MXPA00012554A - Antithrombotic compounds - Google Patents

Abstract

The present invention relates to compounds of formula (I), wherein R1 is phenyl, naphthyl, 1,2,3, 4-tetrahydronaphthyl, (iso)quinolinyl, tetrahydro(iso)quinolinyl, 3,4-dihydro-1H-isoquinolinyl, chromanyl or the camphor group, which groups may optionally be substituted with one or more substitutents selected from (1-8C)alkyl or (1-8C)alkoxy;R2 and R3 are independently H or (1-8C)alkyl;R4 is (1-8C)alkyl or (3-8C)cycloalkyl;or R3 and R4 together with the nitrogen atom to which they are bonded are a nonaromatic (4-8)membered ring optionally containing another heteroatom, the ring optionally being substituted with (1-8C)alkyl or SO2-(1-8C)alkyl;Q is a spacer having a chain length of 10 to 70 atoms;and Z is a negatively charged oligosaccharide residue comprising two to six monosaccharide units, the charge being compensated by positively charged counterions;or a pharmaceutically acceptable salt thereof or a prodrug thereof. The compounds of the invention have antithrombotic activity an can be used in treating or preventing thrombin-related diseases.

Description

ANTITROMBOTIC COMPOUNDS DESCRIPTION OF THE INVENTION The invention relates to novel antithrombotic agents, a process for their preparation, pharmaceutical compositions containing the compounds as active ingredients, as well as the use of said compounds for the manufacture of medicaments. Serine proteases are enzymes that play an important role in the blood coagulation cascade. The members of this protease group are, for example, thrombin, trypsin, Vlla, IXa, Xa, Xla, Xlla, and protein C. Thrombin is the final serine protease enzyme in the coagulation cascade. The main function of thrombin is the cleavage of fibrinogen to generate fibrin monomers, which are entangled to form an insoluble gel. In addition, thrombin regulates its own production through the activation of factors V and VIII above in the cascade. It also has important actions at the cellular level, where it acts on specific receptors to cause platelet aggregation, endothelial cell activation and fibroblast proliferation. In this way, thrombin has a central regulatory role in hemostasis and thrombus formation. Since thrombin inhibitors can have a wide variety of therapeutic applications, an extensive search is carried out in this area. Another important serine protease, factor Xa catalyzes the conversion of prothrombin to thrombin. In the development of synthetic serine protease inhibitors, and more specifically of thrombin, the benzamidine moiety is one of the key structures. This resembles the protonated side chain of the basic amino acids Arg and Lys of their natural substrates. Compounds with this portion have been studied extensively and repeatedly A very potent representative point of this type of thrombin inhibitors is the amino acid derivative Na- (2-naphthylsulfonyl) -glycyl-4-amidinophenylalanine piperidine (NAPAP) (Stürzebecher, J. and others, Thromb Res. 29,635-642, 1983). However, the NAPAP profile is not attractive for therapeutic applications: for example, the compound has a low specific thrombin character and is poorly soluble. The NAPAP derivatives were subsequently prepared, such as the N-alkyl substituted derivatives described in EP 0,236,163, or the glycopeptide derivatives described in EP 0,558,961, Proc. Am. Pept. Symp., 13, h. (60LXAW); 94; pp. 643-5 (Stúber, W. et al., Pept.:Chem., Struct., Biol.,), Proc. Int Symp. Controlled Relay Bioact. Mater. (PCRMEY, 10220178); Vol. 21 st; pp. 712-12 (Walter, E. and others), and EP 0,513,543. However, although these derivatizations may have led to improvements in the pharmacological profile when compared to NAPAP, all these NAPAP-derived compounds remain only active as direct thrombin inhibitors and have a restricted plasma half-life and rapid elimination (from this ma was ^^^ ff ^ ^^^^ Si ^^^^^^^^^^. ^^ ---- ^^ -í ^ ugly ^ ^ ^ representing its anti-thrombin activity only for a short period). It has now been found that the compounds of the formula (I) Z I Q R1-S (0) 2-N (H) -C (H) -C (0) -N (R wherein R is phenyl, naphthyl, 1, 2,3,4-tetrahydronaphthyl, (iso) quinolinyl, tetrahydro (iso) quinolinyl, 3,4-dihydro-1H-isoquinolinyl, chromanyl, or the camphor group, said groups Optionally they can be substituted with one or more substituents selected from alkyl of 1 to 8 carbon atoms or alkoxy of 1 to 8 carbon atoms; R2 and R3 are independently H or alkyl of 1 to 8 carbon atoms; R 4 is alkyl of 1 to 8 carbon atoms or cycloalkyl of 3 to 8 carbon atoms; or R3 and R4 together with the hydrogen atom to which they are attached, are a non-aromatic 4- to 8-membered ring, optionally containing another heterogeneous atom, the ring being optionally substituted with alkyl of 1 to 8 carbon atoms or SO2-alkyl of 1 to 8 carbon atoms; Q is a separator having a chain length of 10 to 70 atoms; and Z is a negatively charged oligosaccharide residue comprising from 2 to 6 monosaccharide units, the charge being compensated through positively charged counterions; or a pharmaceutically acceptable salt thereof or a prodrug thereof which are potent and highly versatile antithrombotic. The compounds of the invention have anti-thrombin activity, but also the structure of the compounds can be modified selectively, so that they have a harmonizable mixed profile of both the non-mediated anti-thrombin (non-mediated factor) activity and the activity anti-Xa mediated by anti-thrombin III (AT-III). The compounds of the invention in this manner are double inhibitors. The compounds of the invention have a long half-life in the plasma and, as a result, have a prolonged anti-thrombin activity compared to NAPAP or its previously reported derivatives. In addition, the compounds of the invention can escape the neutralizing action of platelet factor 4 (PF4). Low toxicity is also an advantageous aspect of the compounds of this invention. Another type of double inhibitors is described in EP 0,649,854. Contrary to the compounds of the present invention, the conjugated saccharide compounds described therein display an anti-thrombin activity mediated by indirect AT-111, in addition to the anti-Xa activity mediated by AT-III. The compounds of the present invention are useful for treating and preventing diseases mediated by thrombin and associated with thrombin. This includes a number of thrombotic and prothrombotic states where the coagulation cascade is activated, which include, but are not limited to, deep vein thrombosis, pulmonary embolism, thrombophlebitis, arterial occlusion of thrombosis or embolism, arterial reocclusion during or after of angioplasty or thrombolysis, restenosis after arterial damage or invasive cardiological procedures, thrombosis or venous embolism after operations, acute or chronic atherosclerosis, shock or attack, myocardial infarction, cancer and metastasis and neurodegenerative diseases The compounds of the invention can also to be used as anticoagulants in extracorporeal blood circuits as needed in dialysis and surgery. The compounds of the invention can also be used as anticoagulants in vitro. The mixed profile of the compounds of the invention can be harmonized by varying the nature of the oligosaccharide residue Z and the length of the Q separator. In this way, a variety of profiles are available. Any negatively charged oligosaccharide residue of 2 to 6 saccharide units can be used in the compounds of the present invention. The suitable compounds of the invention are ^^^^^^^^^^ g ^^ g ^^^^^^^ g? The compounds in which Z is a sulphated or phosphorylated oligosaccharide residue are present. Preferably, the oligosaccharide residue Z is derived from an oligosaccharide, which per se has anti-Xa activity mediated by AT-III, such as the saccharides described in EP 0,454,220 and EP 0,529,715. Particularly preferred oligosaccharide residues are pentasaccharide residues. Most preferably, Z has the formula (II) wherein R5 is independently OSO3"or alkoxy of 1 to 8 carbon atoms Other preferred compounds of the invention are compounds of the formula I, wherein R1 is phenyl, 4-methoxy-2,3,6-trimethylphenyl or naphthyl. In the preferred compounds, NR3R4 represents the piperidinyl group, Preferably, the R2 group is H. The chemical structure of the separator is of minor importance or of no importance for the anti-thrombotic activity of the compounds of the invention, however, it may not be completely rigid, highly flexible separators are more suitable than others, and for synthetic reasons, some separators are more «, -a.e. ^ * -. appropriate than others Suitable separators that can be used easily have, for example, formula III: - [(CH2) 2O] m - [(CH2) n-NR3-C (O) pW- (CH2) 5- (III), where W is - [1,4-phenylene-NR3-C (O) q- (CH2) rS- (CH2) t-S- (CH2) u- [O (CH2) 2] v-O- (CH2) w-C (O) -NR3-; and R3 is independently H or alkyl of 1 to 8 carbon atoms; m = 1-12; n = 1-8; p = 0-4; q = 0 or 1; r = 1-8; s = 1-8; t = 1-8; u = 1-8; v = 1-12; w = 1-8; the total number of atoms is 10-70 atoms, and the portion - [(CH2) 2O] m- is the end with which Q is attached to Z. Preferred separators are the following: - [CH2) 2O] 5 - (CH2) 2-NH-C (O) -CH2-S-CH2-; - [CH2) 2O] 5- (CH2) 2-NH-C (O) -CH2-S- (CH2) 2- [O (CH2) 2] 3-O-CH2-C (O) -NH- ( CH2) 4-, and - [CH2) 2O] 3- (CH2) 2-NH-C (O) -1,4-phenylene-NH-C (O) -CH2-S-CH2-. In the description of the compounds of formula I, the following definitions are used. The term alkyl of 1 to 8 carbon atoms represents a branched alkyl group having 1-8 carbon atoms, for example, methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, hexyl and octyl. Preferred alkyl groups are methyl and ethyl.

The term "alkoxy" of 1 to 8 carbon atoms represents an alkoxy group having from 1 to 8 carbon atoms, the alkyl portion having the previously defined meaning. Methoxy is the preferred alkoxy group. The term cycloalkyl of 3 to 8 carbon atoms represents a cycloalkyl group having 3-8 carbon atoms, being cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl. Preferred cycloalkyl groups are cyclopentyl and cyclohexyl. The length of the separator is the number of atoms of the separator, represented along the shortest chain between Z and the peptide part of the molecule, not representing the oxygen atom of the oligosaccharide Z, which is connected to the separator. The term "prodrug" represents a compound of the invention wherein the amino group of the amidino moiety is protected, for example, by hydroxy or an alkoxycarbonyl group of 1 to 6 carbon atoms. The compounds of the present invention are prepared by derivatizing NAPAP (or a NAPAP analog) at the position of glycine with cysteine or lysine, using methods generally known in the art, said compound subsequently (a) being coupled to an oligosaccharide spacer residue or (b) is coupled to a separator, which is then derivatized with a thiol group and subsequently coupled to an oligosaccharide residue. Any suitable oligosaccharide can be used for this purpose, for example oligosaccharides known in the literature (for example EP 0,454,220 and EP 0,529,715, but not limited to these sources) or commercially available oligosaccharides. Oligosaccharides can be phosphorylated at an appropriate time through methods described, for example, by Bujisman, R, et al., (Abstracts of papers, 9th European Carbonhydrate Symposium Utrecht 1997, Abstract A150). The coupling of the separator to the oligosaccharide can be carried out, for example, using the methods described in EP 0,649,854. Peptide coupling, a method step in the method described above for preparing the methods of the invention, can be performed by methods commonly known in the art for coupling, or condensation, of peptide fragments such as through the azide method , mixed anhydride method, activated ester method, or, preferably, through the carbodiimide method, especially with the addition of catalytic and racemization suppressor compounds such as N-hydroxysuccinimide and N-hydroxybenzotriazole. A summary is presented in The Peptides, Analysis, Synthesis, Biology, Vol. 3, E. Gross and J. Meienhofer, eds. (Academic Press New York, 1981). The amine functions present in the compounds can be protected during the synthetic process through an N-protecting group, which represents a group commonly used in the peptide technique for the protection of a-amino groups, such as the tertiary group. butyloxycarbonyl (Boc), the benzyloxycarbonyl group (Z), the 9-fluoren? lmet? lox? carbonyl group (Fmoc) or the phthaloyl group (Phth). The removal of protective groups can occur in different forms, depending on the nature of those protecting groups. Usually, deprotection occurs under acidic conditions and in the presence of scavengers. A summary of amino protecting groups and methods for their removal is presented in the aforementioned document The Peptides. Analysis, Synthesis, Biology, Vol 3. The compounds of the invention, which may occur in In the form of a free base, they can be isolated from the reaction mixture in the form of a pharmaceutically acceptable salt. Pharmaceutically acceptable salts can also be obtained by treating the free base of formula I with an organic or inorganic acid, such as HCl, HBr, Hl, H2SO, H3PO4, acetic acid, propionic acid, glycolic acid, maleic acid, malonic acid, methanesulfonic acid, fumaric acid, succinic acid, tartaric acid, citric acid, benzoic acid, ascorbic acid and the like. The compounds of this invention possess chiral carbon atoms, and, therefore, can be obtained as an enantiomer pure, or as a mixture of enantiomers, or as a mixture containing diastereomers. Methods for obtaining the pure enantiomers are well known in the art, for example, crystallization of salts which are obtained from optically active acids and the racemic mixture, or chromatography using chiral columns. For diastereomers, they can be used Straight phase or reverse phase columns The compounds of the invention can be administered enterally or parenterally. The exact dose and regimen of these compounds and their compositions will necessarily depend on the needs of the individual to whom the medication is to be administered, the degree of distress or need and the judgment of the physician. In general, parenteral administration requires lower doses than other methods of administration, which are more dependent on absorption. However, daily doses, for Human beings are preferably 0.001-100 mg per kg of body weight, most preferably 0.01-10 mg per kg of body weight. The medicament made with the compounds of this invention can also be used as an aid in therapy acute anticoagulant. In such a case, the medicament is administered with other compounds useful for treating said disease states. Mixed with pharmaceutically suitable auxiliaries, for example, as described in the standard reference, Gennaro and others, Remington's Pharmaceutical Sciences, (18lh ed., Mack Publishing Company, 1990, see especially part 8: Pharmaceutical Preparations and their manufacture) compounds can be compressed into solid dose units, such as pills, tablets, or can be processed in capsules or suppositories. TO Through pharmaceutically acceptable liquids, the compounds they may also be applied in the form of a solution, suspension, emulsion, for example, to be used as a preparation for injection, or as a spray, for example, to be used as a nasal spray or spray. To make dose units, e.g., tablets, the use of conventional additives such as fillers, colorants, polymeric binders, and the like is contemplated. In general, any pharmaceutically acceptable additive that does not interfere with the function of the active compounds can be used. Suitable carriers which can be administered with the compositions include lactose, starch, cellulose derivatives and the like, or mixtures thereof, and used in suitable amounts. The invention is further illustrated through the following examples.

EXAMPLES Abbreviations used: DMAP = N, N-dimethylaminopyridine TEA = triethylamine Z = benzyloxycarbonyl Ac = acetyl MMTr = monomethoxytrityl Bn = Benzyl DCHA = dicyclohexylammonium EDCI = 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride HOBt = 1-hydroxybenzotriazole DiPEA = diisopropylethylamine Pyr = pyridinyl TEG = tetraethylenic glycol HEG = hexaethylenic glycol APA = amidinophenylalanine Cys = cysteine The numbers of the compounds refer to the compounds in the formulas sheets. 10 Trichloroacetamidate of 4-0- (4-0 (2,3,4,6-tetra-0-acetyl-aD-glucopyranosyl) -2,3,6-tri-0-acetyl-aD-glucopyranosyl) -2, 3,6-tri-0-a / β-D-glucopyranosyl (4) To a stirred solution of maltotriose (1) (2.0 g, 4.0 mmol) in 100 ml of pyridine was added acetic anhydride (6.2 ml, 65 mmol) and a catalytic amount of DMAP (0.79 g, 65 mmol;). After 5 hours, the reaction mixture was poured into aqueous hydrogen acid carbohydrate (1 M, 250 ml) and extracted three times with 200 ml of ethyl acetate. The organic layers were dried over MgSO 4 and concentrated in vacuo. The product was purified by column chromatography (light petroleum / ethyl acetate 1/1 to 0/1, v / v) giving 2 as a white foam (91% yield, 3.5 g). Anomeric deacetylation was achieved through the treatment of 2 (3.0 g, 3.1 mmol) with 0.1 M of a solution of hydrazine acetate in dimethylformamide (34 ml, 3.4 mmoles) for 1 hour After concentration in vacuo, the reaction mixture was diluted with 50 ml of ethyl acetate, washed with sodium hydrogen carbonate (1 M, 3 x 25 ml), dried (sulfate and magnesium) and concentrated. Purification through silica gel column chromatography (light petroleum / ethyl acetate, 3/2 to 1/0, v / v) gave 3 (92% yield, 2.7 g). Compound 3 (2.7 g, 3.1 mmol) was diluted in 15 ml of dichloromethane and 1.7 ml of trichloroacetonitrile together with a catalytic amount of cesium carbonate (0.2 g, 0.62 mmol), which was added. After 1 hour the reaction mixture was filtered and the filtrate was concentrated under reduced pressure. Purification of crude 4 through column chromatography (light petroleum / ethyl acetate / TEA, 50/49/1 to 0/99/1, v / v / v) yielded 4 pure as a white foam (1.9 g, 71%). 4-0- (4-0- (2,3,4,6-tetra-0-acetyl-aD-glucopyranosyl-2,3,6-tri-0-acetyl-aD-glucopyranosyl) -2,3 , 6-tri-acetyl-β-D-glucopyranoside of glycol N-benzyloxycarbonyl-1-aminohexamethylene (6) A solution of donor 4 (0.69 g, 0.76 mmol) and acceptor 5 (0.31 g, 0.76 mmol) ) in 1.5 ml of dichloromethane was stirred for 1 hour under a flow of argon in the presence of activated molecular sieves 4a (250 mg) .The solution was cooled to -20 ° C and a solution of 15 μL of trifluoromethanesulfonate was added dropwise. tri-methylsilyl in 0.6 ml of dichloromethane to the reaction mixture After 10 minutes, analysis by TLC (5% methanol in dichloromethane) showed the presence of a product, 03 g of solid sodium acid carbonate was added to the The reaction mixture was stirred for 10 minutes and then filtered, the filtrate was diluted with 50 ml of dichloromethane, subsequently washed with aqueous sodium hydrogen carbonate (1M, 2 x 25 ml) dried (sulfa). magnesium) and concentrated in vacuo. The residue was chromatographed on silica gel (0-4% in ethyl acetate) yielding 6 (0.57 g 56% yield). 4-0- (4-0- (α-D-glucopyranosyl) -a-D-glucopyranosyl) -β-D-glucopyranoside glycol N-benzyloxycarbonyl-1-aminohexaethylenic acid (7). Compound 6 (0.57 g, 0.43 mmol) was treated with a solution of potassium tert-butylate (43 mg, 10 mg per mmol Ac) in 15 ml of methanol. After 1 hour, analysis by TLC (ethyl acetate / pyridine / acetic acid / water, 5/7/4 / 1.6, v / v / v / v) indicates a complete conversion of 6 to 7. The reaction it was neutralized with the ream Dowex 50 WX4-H +. The resin was removed by filtration and the filtrate was concentrated under reduced pressure to provide 7 (0.37 g, 95% yield), which was used without further purification. 4-0- (4-0- (α-D-glucopyranosyl-2,3,4,6-tetrakis- (dibenzylphosphate) -aD-glucopyranosyl-2,3,6-tris (dibenzylphosphate)) - ß-D-glucopyranoside 2 3,6-tris (dibenzylphosphate) glycol N-benzyloxycarbonyl-1-aminohexaethylenic acid (9) A solution of 1H-tetrazole (54 mg, 0.77 mmol) in 1 ml of acetonitrile was added to a mixture of 7 (86 mg , 95 μmol) and 8 (450 mg, 1.4 mmol) in acetonitrile / hexane (2/1, v / v, 2 ml) After stirring for 1 hour at 20 ° C, the reaction mixture was cooled with a bath of ice and 0.75 ml of ter-bitylhydroperoxide were added.The stirring was continued for 45 minutes, after which the analysis by TLC showed the presence of a main product.The purification through silica gel chromatography (100/0 to 95/5 dichloromethane / methanol, v / v) produced pure 9 (311 mg, 92% yield). 4-0- (4-0- (aD-glucopyranosyl 2,3,4,6-tetrakis phosphate) -aD-glucopyranosyl 2,3,6-triphosphate) -β-D-glucopyranoside 2,3,6-triphosphate 1-aminohexethylene glycol (10). Compound 9 (311 mg, 87 μmoles) was dissolved in tert-butanol / water 6/1, v / v, 20 ml) containing a few drops of acetic acid. The solution was stirred under a continuous stream of hydrogen in the presence of 10% Pd / C (100 mg). After 3 hours, the Pd / C catalyst was removed by filtration and the filtrate was concentrated in vacuo. Then the Dowex 50 WX4-Na + ion exchange produced 10 (179 mg, 98% yield).

N-2-naphthalene sulfonyl-S-4-monomethoxytrityl- (L) -cysteine (12). To a stirred mixture of commercially available S-4-monomethoxytrityl- (L) -cysteine (11) (0.34 g, 1 mmol), 5 ml of dioxane and 5 ml of 10% sodium carbonate, was added sodium chloride. Naphthalenesulfonyl (0.25 g, 11 mmol) After stirring for 1 hour, the reaction mixture was acidified through the addition of 50 ml of 5% aqueous citric acid and extracted with ethyl acetate (2 x 50 ml. ). The combined organic layers were dried (magnesium sulfate) and concentrated under reduced pressure. The crude product was chromatographed on silica gel (methanol / dichloromethane / triethylamine, 0/99/1 to 4/95/1, v / v / v) to yield 12 (36% yield, 0.44 g).

N-2-naphthalenesulfonyl-S-2-pyridinesulfenyl- (L) -cysteine (14). A solution of trifluoroacetic acid and triisopropylsilane in dichloromethane (1/1/18, v / v / v) was added to compound 12 (0.44 g, 0.76 mmol). After stirring for 20 minutes, the mixture was emptied with water and extracted with dichloromethane (2 x 50 ml). The layers The combined organics were dried over magnesium sulfate and concentrated in vacuo. The traces of trifluoroacetic acid in the crude mixture were removed by co-evaporation with toluene. The resulting free thiol 13 was redissolved in 2.5 ml of isopropanol and a solution of Aldrithiol ™ (1.7) was added dropwise. g, 7.6 mmol) in isopropanol / 2 N aqueous acetic acid (1/1, v / v, 20 ml). After 1 hour, analysis by TLC indicated that the reaction was complete and the mixture was concentrated under reduced pressure. Traces of acetic acid in the residue were removed by co-evaporation with toluene. The crude product was dissolved in ml of acetone and to this solution 0.3 ml of ^ * s¡ and ». Then, the compound 14 was precipitated from the reaction mixture as its DCHA salt. The precipitate was isolated, dissolved in 50 ml of ethyl acetate and washed with 5% aqueous citric acid (2 x 30 ml). The organic layer was dried (magnesium sulfate) and concentrated under reduced pressure to give pure 14 (55% yield, 0.25 g) N (N (Na-2-naphthalenesulfonyl-S-2-pyridinesulfenyl- (L) -cysteinyl) - (D, L) -4-am idinofen i I to nyl) piperidine (16). To a solution of N - ((D, L) -4-amidinophenylalanyl) p? Peridine dichlorohydrate (15) (0.13 g, 0.39 mmol) and the cysteine derivative 14 (0.16 g, 0.39 mmol) in 2 ml of dimethylformamide HOBt (58 mg, 0.42 mmol), EDCI (82 mg, 0.42 mmol) and N-ethylmorpholine (110 μL, 0.78 mmol) were added. After stirring for 16 hours, the mixture was diluted with 20 ml of dichloromethane and washed with water (2 x 19 ml). The organic layer was dried (magnesium sulfate) and concentrated in vacuo. The residue was purified through silica gel column chromatography (first 10-20% methanol in dichloromethane to remove impurities and then ethyl acetate / pyridine / acetic acid / water (16/7 / 1.6 / 4, v / v / v / v) to release the product), and subsequently through gel filtration on Sephadex LH-20 (eluent: methanol / dichloromethane, 4/1, v / v) to produce homogeneous 16 (70%, 0.19 g ).

. ^^^ ^ - ^ ~ • J-fÜÉ fttJMtilfiMt ^ - '^ ^^^ ífe ^. ^^ ^? - ^. ^^ j5 ^^ 3- ^ £ jgE ^. ^. , - ... .-. -S »..« A. * - & Conversion coupling of maltotriose decaphosphate 18 with peptide 16. To a solution of maltotriose decaphosphate 18 (21 mg, 9.8 pmol) in a pH regulator of 0.1 M Na2HPO4 (1.0 ml pH 7.5) was added a solution of N-hydroxysuccinimidyl 2-bromoacetate in 1 ml of ethanol. After stirring for 2 hours, the reaction mixture was applied on a Sephadex G25 column eluted with 10% acetonitrile in water. The appropriate fractions were combined and concentrated under reduced pressure at low temperature (25 ° C) to produce compound 19. The NAPAP 16 analogue (10 mg, 15 μmol) was dissolved in a mixture of 1 ml of methanol and the pH regulator 0.1 M Na2HPO4 (0.75 ml, pH 7.0) degassed by passing it through helium and through sound application. To this solution, tributylphosphine (4.1 μL, 16 μmol) and the mixture of The reaction was stirred under an argon atmosphere. After 2 hours, analysis by HPLC (Lichrospher® RP18-column) indicated a complete cleavage of the 2-pyridinesulfenyl group and a solution of compound 19 in 0.25 ml of dimethylformamide and pH regulator 0.1 M Na2HPO4 was added to the reaction mixture. (0.50 ml, pH 7.0).

The mixture was stirred for 3 hours, after which the crude mixture was purified by gel filtration (Fractogel HW-40, eluent: 0.15 M TEAB). The concentration of the appropriate fractions and the subsequent exchange of Dowex 50 WX-Na + ions provided, after lyophilization, a homogeneous conjugate I (10.1 mg, 47% yield). The two diastereoisomers were ^ a - ^ - ~ ^^^^ »** ^? - TllÍÍ It- > - - "--A '" r.?^-^^aaEáia^^ separated through semi-preparative HPLC column chromatography (LiCrospher RP-18 column, gradient: 17.5% -22.4% CH3CN in 0.05 M TEAA aqueous) to give the diastereoisomer Ia (retention time: 28.6 minutes) and diastereoisomer lb (retention time: 33.0 minutes). The two isomers were desalted through gel filtration (Sephadex G-25 DNA, superfine grade), transformed to the Na + - form using resin and Dowex 50 WX-Na + ion exchange. Diastrereoisomer la: 1H NMR (D2O, 600 MHz, HH-COZY): maltotriose: (reduction end) 4.65 (bs, 1H, H1), 3.85 (m, 1H, H2), 4.35 (m, 1H, H3), 3.76 (m, 1H, H4); 5.50 (bs, 1H, H1 '), 4.18 (rn, 1H, H2'), 4.10 (m, 1H, H4 '); (without reduction end) 5.71 (bs, 1H, H1"), 4.09 (n-, 1 H, H2"), 4.45 (m, 1H, H3"), 4.15 (m, 1 H, H4"); 3.95-3.84 (H5, maltotriose); separator: 3.65-3.51 (m, 22H, OCH2HEG), 3.35 (m "2H, CH2NH2), 3.15 (s, 2H, SCH2 (O)); peptide: 8.31 (s, 1H, HaromNAS), 8.06-7.67 (n-, 6H, HaromNAS), 7.70, 7.17 (2 xd, 4H, Harom APA, J = 7.8 Hz), 4.28 (m, 1H, aCH APA) , 3.91 (n-, 1 H, aCH Cys), 3.30-3.04 (m, 4H, CH2N piperidine), 2.82-2.62 (m, 311, ßCH2 Cys, ßCH APA), 2.57 (m, 1H, ßCH 'APA) , 1.45-1.25 (m, 6H, CH2 piperidine); ES-MS: [M-3H] 3"724.1, [M-2H] 2" 1086.7. Diaesteroisomer lb: 1 H NMR (D 2 O, 600 MHz, HH-COZY): maltotriose: (reduction end) 3.80 (m, 1 H, H 2), 4.32 (m, 1 H, H 3), 3.89 (m, 1 H, H 4); 5.49 (bs, 1H, H1 '), 4.22 (m, 1H, H2'), 4.11 (m, 1H, H4 '); (without reduction end) 5.70 (bs, 1H, H1"), 4.22 (m, 1H, H2"), 4.52 (m, 1H, H3"), 4.24 (m, 1H, H4"); 3.91-3.84 (H5, maltotriose), separator '363-3 52 (m, 22H, OCH2 HEG), 3 (t, 2H, CH2NH2), 3.17 (AB, 2H, SCH2 (O)); peptide: 8.35 (s, 1H, Haro ?? NAS), 8.07-7.65 (m, 6H, HaromNAS), 7.77, 7.22 (2 xd, 4H, Har? m APA, J = 7.8 Hz), 4.62 (t, 1H , aCH APA, JaCH.ßcH = 7.3 Hz), 4.05 (m, 1H, aCH Cys), 3.05-3.00 (m, 4H, CH2N piperidine), 2.85-2.67 (m, 4H, ßCH2 Cys, ßCH2 APA)), 1.88-1.24 (m, 6H, CH2 piperidine); ES-MS: [M-3H] 3"724.0, [M-2H] 2" 1086.2.

N-Hydroxysuccinimidyl-14-S-2-pyridinesulfenyl-14-mercapto-3,6,9,12-tetraoxatetradecanoate (22). Separator 20 (0.75, 2.4 mmol) (P. Westerduin et al., Angew. Chem. Int. De. Engl. 1996, 35, 3, p 331-333) and Aldrithiol ™ 2.6 g, 12.1 mmol) was dissolved in 20 ml. ml of dichloromethane and treated with 4 ml of n-butylamine. After stirring for 2 hours, the reaction mixture was concentrated in vacuo, redissolved in 50 ml of dichloromethane and washed with 5% aqueous citric acid (2 x 50 ml). The organic layer was dried and concentrated under reduced pressure. Silica gel column chromatography (methanol / acetic acid / dichloromethane, 0/1/99 to 6/1/93, v / v / v) of the residue yielded pure 21 (0.80 g, 88% yield). Compound 21 (0.80 g, 2.1 mmol) was dissolved in 10 ml of dichloromethane and N-hydroxysuccinimide (0.26 g, 2.3 mmol) and EDCI (0.45 mg, 2.3 mmol) were added to this solution. After 1 hour, the reaction mixture was diluted with 50 ml of dichloromethane, washed three times with 20 ml of ice water, dried (magnesium sulfate) and concentrated to give the reaction mixture. & B¡SB ¡i-C.-j tí * »',! (0.98 mg, 98% yield), which was used without further purification.

Ne-tert-Butyloxycarbonyl-Nabenzensulfonyl- (L) -lysine (24). It was prepared as described for 12, using 23 and d-benzensulfonyl chloride as starting materials (0.86 mg, 75% yield).

Ne- (14-s-2-pyrid insult i 1-14-mercapto-2, 6, 9,12- 10 tetraoaxatetradecanol) - Na-benzensulfonyl- (L) -lysine (26). Compound 24 (0.86 g, 2.2 mmol) was treated with 3 N of hydrogen chloride in ethyl acetate. After 15 minutes, the reaction mixture was concentrated in vacuo. The traces of acid in the residue were removed by co-evaporation with toluene. He Crude oil was dissolved in a dioxane / water mixture (4/1, v / v, 2.5 ml) and to this solution was added compound 22 (0.98 g, 2 1 mmol) and DiPEA (1.1 ml, 6.6 mmol). . After 1 hour, the reaction mixture was diluted with 100 ml of dichloromethane and washed with 5% aqueous citric acid (2 x 50 ml). The organic layer dried (Magnesium sulfate) and concentrated in vacuo. The residual oil was purified by silica gel column chromatography (0-10% methanol / ethyl acetate) to give homogeneous 26 (0.95 g, 67% yield).

Fe ^ g¡aj g¡¡ ^ ^ fc 6 ^ N (N (Ne- (14-S-2-Pirinsulfenyl-14-mercapto-3,6,9,12-tetraoxatetradecanoyl) -Na-benzenesulfonyl- (L ) -lysinyl) - (D, L) -4-amidinophenylalanyl) piperidine (27). It was prepared as described for 16, using 26 and 15 as starting materials (87 mg, 70% yield).

Conversion coupling of maltotriose-decaphosphate 18 with peptide 27 (II). Prepared as described for I, using 18 and 27 as starting materials. Purification of crude II was carried out by semi-preparative HPLC (LiChrospher ® RP-18 columns). After desalting through gel filtration (Sephadex G-25 DNA-superfine grade), transformation to the Na + - form using the Dowex 50 WX4-Na + ion exchange resin and lyophilization provided pure II as a fluffy solid white (8.5 mg, 23% yield from 18). H NMR (D2O, 600 MHz, HH-COZY): maltotriose: (reduction end) 4.67 (m, 1H, H1), 4.07 (m, 1H, H2), 4.40 (m, 1H, H3), 4.06 (m , 1H, H4); 5.49 (bs, 1H, H1 '), 4.24 (m, 1H, H2'), 4.66 (m, 1H, H31), 4.10 (m, 1H, H4 '); (without reduction end) 5.73 (bs, 1H, H1"), 4.17 (m, 1H, H2"), 4.3 8 (m, 1H, H3"), 4.22 (m, 1H, H4"); 3.95-3.82 (H5, maltotriose); separator: 4.03, 4.02 (2 xs, 2H, OCH2C (O)), 3.73-3.59 (m, 36H, OCH2 TEG, HEG), 3.38 (t, 2H, CH2NH2), 3.27, 3.26 (2 xs, 2H, SCH2 (O)), 2.75 (2 xt, 2H, CH2S); peptide: 7.81-7.52 (m, 5H, HaromBS), 7.79, 7.76, 7.42, 7.41, (4 xd, 4H, HaromAPA), 4.87, 466 (2 xt, 1H, aCH APA, JucH PCH = 4 Hz), 407 (m, 1H, aCH Lys), 3.37-3.73 (m, 8H, eCH2 Lys, ßCH2 APA, CH2N piperidine), 1.96-1.46 (m, 12H, CH2 piperidine, ß / ?, dCH2 Lys); ES-MS [M + 3 H] 3+ 801.2, [M + 2 H] 2 + 12008. 5 Pentasaccharide 30 partially protected. The known pentasaccharide 29 (53 mg, 49 μmoles) (R.C. Bujisman et al., Chem. Eur. J 1996, 2, 12, p 1572-1577) was dissolved in 0.25 ml of dimethyl formamide and 1 ml of water and treated with N- (benzyloxycarbonyloxy) -succinimide (18 mg, 72 μmoles) and 28.6 ml of N-ethylmorpholine. After stirring for 15 minutes, analysis by TLC (ethyl acetate / pyridine / acetic acid / water, 5/7 / 1.6 / 4, v / v / v / v) revealed that the reaction was complete and the reaction mixture was applied directly on a column of RP-18, which was eluted with water-ethanol (90/10 to 60/40). The appropriate fractions were combined and concentrated to a small volume and applied on a Dowex 50 WX4-H ion exchange column in water. The eluted product was concentrated in vacuo to yield pure (54 mg, 91% yield). 20 Sulfated Pentasaccharide 32. Compound 30 (54 mg, 45 μmol) was dissolved in 1 ml of dimethylformamide. The complex of triethylamine sulfur dioxide (0.51 g, 5 equivalents for each hydroxyl group) was added, and the mixture was stirred under a nitrogen atmosphere at 55 ° C for 16 hours. The mixture was subsequently cooled to 0 ° C and aqueous sodium hydrogen carbonate (5 equivalents for each equivalent of the triethylamine sulfothioxide complex) was added. The mixture was stirred for 1 hour, concentrated to a small volume and applied on a Sephandex G-25 column, which was eluted with 10% acetonitrile in water. The appropriate fractions were combined and concentrated to a small volume, which was subsequently passed through a column of Dowex 50 WX4 (Na + form) eluted with water. The eluted product was concentrated and redissolved in 1 ml of 0.2 N hydrogen chloride and allowed to stand for 16 hours at 4 ° C. The reaction mixture was neutralized with 0.1 sodium hydroxide and desalted on a Sephadex G-25 column and eluted with 10% acetonitrile in water to provide homogeneous. Compound 31 was dissolved in tert-butanol / water 6/1, v / v, 20 ml) containing a few drops of acetic acid. The solution was stirred under a stream of hydrogen in the presence of 10% Pd / C (100 mg). After 3 hours, the Pd / C catalyst was removed by filtration and the filtrate was concentrated in vacuo to yield pure 32 (60 mg, 60% yield).

Coupling by condensation of pentasaccharide 32 with peptide 16. Pentasaccharide 32 (15 mg, 6.5 μmoles) was dissolved in pH buffer of 0.1 M NaH2PO (2 ml, pH 7.5) and to this solution was added sulfo-SlAB ™ (16 mg, 33 μmol) After stirring for 3 hours in the dark, analysis by HPLC (monoQ anion exchange) revealed that the reaction was complete and the crude compound 34 was purified on a Superdex 30 column (10% acetonitrile in Water). The appropriate fractions were combined and concentrated in vacuo at low temperature (25 ° C). To this solution of the analogue NAPAP 16 (9 mg, 14 μmol) in a mixture of 1 ml of methanol and pH regulator of 0.1 M Na2HP04 (0.75 ml, pH 7.0) degassed by passing it through helium and by applying sound before of use, tributylphosphine (3.9 μL, 15 μmol) was added. After stirring for 1 hour under an argon atmosphere, analysis by HPLC (Lichrospher® RP-18 column) indicated a complete cleavage of the 2-pyridinesulfenyl group. A solution of the derivatized pentasaccharide 34 0.25 ml of dimethylformamide and pH regulator of 0.1 M Na2HP0 (0.50 ml, pH 7.0) was added, and the mixture was stirred for 3 hours. The crude product was applied on a Sephadex G-50 column, which was eluted with 10% acetonitrile in water. The appropriate fractions were combined, concentrated to a small volume and desalted on a Superdex 30 column, which was eluted with 10% methanol in water. Concentration and lyophilization yielded conjugate III as a white solid (9 mg, 52% yield). 1 H NMR (D 2 O, 600 MHz, HH-COZY): d 3.60, 3.53, 3.43 (3 x s, 9H, CH 3 OE, G, H); D. 5.53 ring (m, 1H, H1), 4.15 (m, 1H, H2), 4.58 (m, 1H, H3), 3 56 (m, 1H, H4), 392 (m, 1H, H5), 4 26, 4 13 (2 xm, 2H, H6, H6 '); ap / 7 /? £: 4.70 (d, 1H, H1, J ?, 2 = 8.1 Hz,), 4.21 (m, 1H, H2), 3.62 (m, 1H, H3), 3.92 (m, 1H, H4), 3 74 ( m, 1H, H5); an / 7 / o F: 5.39 (d, 1H, H1, J?, 2 = 38 Hz), 4.22 (m, 1H, H2), 4.56 (m, 1H, H3), 3.83 (t, 1H, H4, J3,4 = J4, s = 9.8 Hz), 4.12 (m, 1H, H5), ap / 7 / or G: 5.15 (bs, 1H, H1), 4.35 (m, 1H, H2), 3 76 (m , 1H, H3), 421 (m, 1H, H4), 4.80 (m, 1H, H5); ring H: 5.10 (d, 1H, H1, JL2 = 3 .6 Hz), 4.31 (m, 1H, H2), 4.54 (m, 1H, H3), 4.21 (m, 1H, H4); separator: 7.51, 7.53, 7.13, 712 (4 x d, 4H, HaromSIAB), 3.73 (m, 2H, CH2CH2NH2), 3.66 (m, 12H, OCH2 TEG), 3.31 (m "2H, CH2NH2); Peptide: 8.27, 8.22 (2 xs, 1H, HaromNAS), 7 98-760 (m, 6H, HaromNAS), 7.71, 7.64, 7.46, 7.44 (4 xd, 4H, HaromAPA), 4.60, 4.45 (2 xt, 1H , aCH APA, JacH, ßcH = 6.6 Hz), 4.00, 3.97 (2 xm, 1H, aCH Cys), 3.10-2.85 (m, 4H, CH2N piperidine), 2.822.70 (m, 3H, ßCH2 Cys, ßCH APA ), 2.61 (m, 1H, ßCH 'APA), 1.55- 15 1.15 (m, 6H, CH2 piperidine); ES-MS. [M-H] "2680.6 Using similar methods, the following compounds were prepared: ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ f ^^^^^^^^^^^^ ^^^^^^^^^^^^^^^^ g ^ r •? in1riir - «^« «^ - - ^« ^ «- ^ ---' - .- ^ '- * - ~ -a-fa» fci -.- J. _to? --- »- - £" * "**** -" »'» * • - ~ ^ - > * ^ * J- ^., -J - ^ - ^, - ^ i .. ^. ^ ..,.

The biological activities of the compounds of the present invention were determined through the following test methods.

I. Anti-thrombin assay. Thrombin (factor lia) is a factor in the coagulation cascade. The anti-thrombin activity of the compounds of the The present invention was analyzed by spectrophotometrically measuring the rate of hydrolysis of chromogenic substrate s-2238 exerted by thrombin. This assay for anti-thrombin activity in a pH regulator system was used to determine the IC50 value of a test compound. 15 Test Medium: Trometamine-NaCl-glycol pH regulator iajjja t | ^ j¡ | jK) | fitt | ¡gi ||| £ iHHB ^ -to-S .. ..- ^ ^^ ¡ßk & & polyethylenic 6000 (TNP) Reference compound: 12581 (Kabi). Vehicle: TNP pH regulator. The solubilization can be assisted with dimethyl sulfoxide, methanol, ethanol, acetonitop, or tert-butyl alcohol, which do not produce adverse effects in concentrations of up to 2.5% in the final reaction mixture. Technique: Reagents * 1. Trometamine pH regulator- NaCl (TN). PH composition: Trometamine (Tris) 6.057 g (50 mmol) NaCl 5.844 g (100 mmol) Water for 1 1 The pH of the solution was adjusted to 7.4 at 37 ° C with HCl (10 mmol-1"1). TNP pH regulator Polyethylene glycol 6000 was dissolved in the TN pH regulator to give a concentration of 3 g-1"1. 3. Solution S-2238 One vial of S-2238 (25 mg, Kabi Diagnostica Sweden) was dissolved in 20 ml of the TN pH regulator to give a concentration of 1.25 mg-ml "1 2 mmoles-1" +. 4. Thrombin solution Human thrombin (16,000 nKat-vial "1, Central Laboratorium voor Bloedtransfusie, Amsterdam, The Netherlands) ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ ^^^^^^^^^^ was dissolved in pH regulator of TNP to give a supply solution of 835 nKat.m 1. Immediately before use, this solution was distilled with the pH regulator of TNP to give a concentration of 3.34 nKat.ml "1. * -All the ingredients used are of analytical grade.-For the aqueous solutions, ultra pure water was used (Milli-Q quality) .Preparation of the solutions of test and reference compounds. Test and reference compounds were dissolved in Milli-Q water to give supply concentrations of 1 () "2 mol-1. Each concentration was diluted stepwise with the vehicle to give concentrations of 10" 3, 10"4, and 10. ~ 5 mol-1"1. The dilutions, including the supply solution, were used in the assay (final concentrations in the reaction mixture 3-10" 3.10"3; 3-10" 4; 10"4; 3-10"5.10" 5; 3-106 mol-1"1, respectively).

Process. At room temperature, 0.075 ml and 0.025 ml of the solutions of the test compound or the reference compound or vehicle were alternately pipetted into the wells of a microtitre plate, and these solutions were diluted with 0.115 ml and 0.0165 ml of TNP pH regulator, respectively. An aliquot of 0.030 ml of the solution was added ^ - ^.,. ...- jáj »t-¿a¿. ^. ^ »< s ^ & < tJ-a. n > müí? . , S »afe - ^ - 3iii ^ a- ^ ai ^ á ^ < gfe-Av »¡i- ^ Jaafa.fa.-j. , < -.-.- -... jaS- t-jr «an - *. * .- - S-2238 to each cavity, and the plate was preheated and incubated with shaking in an incubator (Amersham) for 20 minutes at 37 ° C. ° C. After preincubation, the hydrolysis of S-2238 was initiated through the addition of 0.030 ml of a thrombin solution to each well. The plate was incubated (with shaking for 30 seconds) at 37 ° C. When starting after 1 minute of incubation, the absorbance of each sample at 404 nm was measured every 2 minutes, for a period of 90 minutes, using a kinetic microtiter plate reader (Twinreader plus, Flow Laboratories). All data was recorded on a personal computer IBM using LOTUS-MEASURE. For each concentration of the compound (expressed in a reaction mixture in mol-1"1) and for the template, the absorbance was plotted against the reaction time in minutes.

Evaluation of responses: for each final concentration, the maximum absorbance was calculated from the test chart. The IC5o value (final concentration, expressed in μmol-1"1, causing a 50% inhibition of the maximum absorbance of the template) was calculated using the logit transformation analysis (logarithmic unit) according to Hafner et al., ( Arzneim-Frosch./Drug Res. 1977; 27 (ll): 1871-3).

Antithrombin activity.

II. Anti-factor Xa assay. Activated factor X (Xa) is a factor in the coagulation cascade. The anti-Xa activity of the compounds of the present invention was determined by spectrophotometrically measuring the hydrolysis rate of the chromogenic substrate s-2222, exerted by Xa. This assay for anti-Xa activity in a pH regulator system was used to determine the IC50 value of the test compound. In general, the following procedure and test conditions were analogous to those of the anti-thrombin assay described above. The differences are indicated below.

Reference compound: benzamidine Vehicle: pH regulator of TNP. Solubilization can be assisted with dimethyl sulfoxide, methanol, ethanol, acetonitrile or tert-butyl alcohol, which have no adverse effects at concentrations of up to 1% (for DMSO) and 2.5% (for the other solvents) in the mixture. final reaction.

Technique: Reagents * 3. solution S-2222 S-2222 (15 mg, Kabí Diagnostica, Sweden) was dissolved in a bottle in 10 ml of water to give a concentration of 1.5 mg-ml "1 2 mmoles-1" 1. 4. Xa solution. Human bovine factor Xa (71 nKat-vial "1; Kcibi Diagnostica) was dissolved in 10 ml of pH regulator of TNP and then further diluted with 30 ml of pH regulator of TNP to give a concentration of 1.77 nKat-ml "1. The dilution has to be freshly prepared Procedure: Instead of the solution of S-2238 (in the anti-thrombin assay), it was added the solution of S-2222 to each cavity in this assay, anti-factor Xa activity. twenty LAMINA FORMULA 1 FORMULA 2 LAMINA LAMINA FORMULA 3 I 11 1888 R K R * = H I- • - 19 R BrAc II LAMIN ORMULA 4 Q, LAMINA FORMULA 5

Claims (10)

1. CLAIMS A compound of the formula (I). Z I Q R'-S (0) 2-N (H) -C (H) -C (0) -N (R wherein R 1 is phenyl, naphthyl, 1,2,3,4-tetrahydronaphthyl, (iso) quinolinyl, tetrahydro (iso) quinolinyl, 3,4-dihydro-1 H-isoquinolinyl, chromanyl, or the camphor group, said groups optionally they can be substituted with one or more substituents selected from alkyl of 1 to 8 carbon atoms or alkoxy of 1 to 8 carbon atoms; R2 and R3 are independently H or alkyl of 1 to 8 carbon atoms; R 4 is alkyl of 1 to 8 carbon atoms or cycloalkyl of 3 to 8 carbon atoms; or R3 and R4 together with the hydrogen atom to which they are attached, are a non-aromatic 4- to 8-membered ring, optionally containing another heterogeneous atom, the ring being ^^^^^^^^^^^^^^^^^^^^^^^^^^. ** - / swirfliia-i &t-teg optionally substituted with alkyl of 1 to 8 atoms carbon or SO 2 -alkyl of 1 to 8 carbon atoms; Q is a separator having a chain length of 10 to 70 atoms; and Z is a negatively charged oligosaccharide residue comprising from 2 to 6 monosaccharide units, the charge being compensated through positively charged counterions; or a pharmaceutically acceptable salt thereof or a prodrug thereof.
2. 2. The compound according to claim 1, wherein Z is derived from an oligosaccharide, which has, per se, an anti-Xa activity mediated by AT-III.
3. 3. The compound according to claim 2, wherein Z is a pentasaccharide residue.
4. 4. The compound according to claim 3, wherein Z has the formula (II) ' wherein R5 is independently OSO3 or alkoxy of 1 to 8 carbon atoms.
5. 5. The compound according to claims 1 to 4, 4? wherein R 'is phenyl, 4-phthate' l-2,3 > 6-tr? Methylene or naphthyl; R2 is H, and NR3R4 represents the piperidinyl group.
6. 6. The compound according to claims 1 to 5, wherein Q has the formula (III). 5 - [(CH2) 2O] m - [(CH2) n-NR3-C (O)] pW- (CH2) 5- (III), wherein W is - [1,4-phenylene-NR3-C ( O) q- (CH2) rS- or - (CH2) tS- (CH2) u- [O (CH2) 2] vO- (CH2) wC (O) -NR3-; 10 and R3 is independently H or alkyl of 1 to 8 carbon atoms; m = 1-12; n = 1-8, p = 0-4; q = 0 or 1; r = 1-8; s = 1-8; t = 1-8; u = 1- 8; v = 1-12; w = 1-8; the total number of atoms is 10-70 atoms; and the portion - [(CH2) 2O] m- is the end with which Q is bonded to Z.
7. 7. The compound according to claim 6, wherein Q is selected from: - [CH2) 2O] 5 - (CH2) 2-NH-C (O) -CH2-S-CH2-; - [CH2) 2O] 5- (CH2) 2-NH-C (O) -CH2-S- (CH2) 2- [O (CH2) 2] 3-O-CH2-C (O) -NH- ( CH2) 4-; and - [CH2) 2O] 3- (CH2) 2-NH-C (O) -1,4-phenylene-NH-C (O) -CH2-S-CH2-.
8. 8. A pharmaceutical composition comprising the compound according to claims 1 to 7 and pharmaceutically acceptable auxiliaries.
9. 9. The compound of any of claims 1 to 7 for use in therapy. 25
10. 10. The use of the compound of any of the ^^^^^^^^^ & ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ diseases related to thrombin The present invention relates to the compounds of the formula I, wherein fif is phenyl, naphthyl, 1, 2,3,4-tetrahydronaphthyl, (iso) quinolinyl, tetra-3,4-dihydro-1H-isoquinolinyl, chromanyl, nfor, said groups can optionally be substituted with one or more substituents selected from alkyl of 1 to 8 carbon atoms or alkoxy of 1 to 8 carbon atoms; R2 and R3 are independently H or alkyl? 1 to 8 carbon atoms; R is alkyl of 1 to 8 carbon atoms or cycloalkyl of 3 to 8 carbon atoms; or R3 and R4 together with the hydrogen atom to which they are attached, are a non-aromatic 4- to 8-membered ring, optionally containing another heterogeneous atom, the ring being optionally substituted with alkyl of 1 to 8 carbon atoms or SO2-alkyl from 1 to 8 carbon atoms; Q is a separator having a chain length of 10 to 70 atoms; and Z is a negatively charged oligosaccharide residue comprising from 2 to 6 monosaccharide units, the charge being compensated through positively charged counterions; or a pharmaceutically acceptable salt thereof or a prodrug thereof. The compounds of the invention have antithrombotic activity and can be used to treat or prevent thrombin-related diseases. l ... .. SiSS btt? mmmm ^ m ^ mm ^ m ^ m. i fíilJÉÉf?