MXPA00005483A - New compounds - Google Patents

Abstract

There are provided compounds of formula (I), wherein R1, R2, R3, Rx, Y, n and B have meanings given in the description which are useful as competitive inhibitors of trypsin-like proteases, such as thrombin, and in particular in the treatment of conditions where inhibition of thrombin is required (e.g. thrombosis) or as anticoagulants.

Description

NEW USEFUL PHARMACEUTICAL COMPOUNDS AS SELECTIVE SERENE PROTEASE INHIBITORS FIELD OF THE INVENTION The present invention relates to novel pharmaceutically useful compounds, in particular selective inhibitors of serine proteases such as trypsin, especially thrombin, their uses as medicaments, pharmaceutical compositions containing them and synthetic routes for their production.

BACKGROUND OF THE INVENTION Blood coagulation is the key process involved in both hemostasis (ie, preventing blood loss from a damaged vessel) as thrombosis (ie, the formation of a blood clot, sometimes leading to vessel obstruction). Coagulation is the result of a complex series of enzymatic reactions. One of the last stages in this series of reactions is the conversion of proenzyme prothrombin to active enzymatic thrombin.

REF; 120265 It is known that thrombin plays a central role in coagulation. These active platelets, which lead to the aggregation of the platelet, convert the fibrogen to fibrin monomers, which polymerize spontaneously in fibrin polymers, and activate factor XIII, which in turn crosslinks the polymers to form insoluble fibrin. In addition, thrombin activates factor V and factor VIII leading to a generation of "positive realization" of thrombin from prothrombin. By inhibiting platelet aggregation and fibrin formation and cross-linking, effective thrombin inhibitors would therefore be expected to exhibit anti-thrombotic activity. In addition, antithrombotic activity would be expected to be increased by the effective inhibition of the positive feedback mechanism.

PREVIOUS TECHNIQUE The early development of low molecular weight thrombin inhibitors has been described by Claesson in Blood Coagul. Fibrinol. (1994) 5, 411. Block et al. (in J. Clin. Lab. Invest. 24, suppl. 107, 59, (1969)) reports thrombin inhibitors. based on the amino acid sequence located around the cleavage or separation site for the fibrinogen chain Aa. The amino acid sequences are discussed, these authors suggest that the tripeptide sequence Phe-Val-Arg (P9-P2-P1, hereinafter referred to as the sequence P3-P2-Pl) would be the most effective inhibitor. Thrombin inhibitors based on dipeptidyl derivatives with an α, β-aminoalkyl guanidine in the Pl position are known from U.S. Patent No. 4,346,078 and International Patent Application WO 93/11152. Structurally related dipeptidyl derivatives, similar, have also been reported. For example, International Patent Application WO 94/29336 describes compounds with, for example, aminomethyl benzamidines, cyclic aminoalkyl amidines and cyclic aminoalkyl guanidines in the Pl position; European Patent Application 0 648 780, describes compounds with, for example, cyclic aminoalkyl guanidines in the Pl position. Thrombin inhibitors based on peptidyl derivatives, which also have cyclic aminoalkyl guanidines (for example either 3- or 4-aminomethyl) -1-amidinopiperidina) in the position Pl are known to from European Patent Applications 0 468 231, 0 559 046 and 0 641 779. Inhibitors. of thrombin based on tripeptidyl derivatives with arginine aldehyde at the P position are first described in European Patent Application 0 185 390. More recently, peptidyl derivatives based on arginine aldehyde, modified at the P3 position, have been reported. For example, International Patent Application WO 93/18060 describes hydroxyl acids, European Patent Application 0 526 877 describes amino acids, and European Patent Application 0 542 525 describes O-methyl mandelic acids in the P3 position. Also known are serine protease inhibitors (for example thrombin) based on electrophilic ketones in the Pl position. For example, European Patent Application 0 195 212 describes α-keto peptidyl esters and amides, European Patent Application 0 362 002 fluoroalkylamide ketones, European Patent Application 0 364 344 composed of a, β, d-triceto and European Patent Application 0 530 167 derivatives of a-alkoxy-arginine ketone in the Pl position.

Other structurally different serine protease inhibitors such as trypsin are known based on C-terminal boronic acid derivatives of arginine and isothiouronium analogs thereof from European Patent Application 0 293 881. More recently, inhibitors have been described. of thrombin based on peptidyl derivatives, and amino acid in European Patent Application 0 669 317 and International Patent Applications WO 95/35309, WO 95/23609, WO 94/29336, WO 97/02284, WO v 97/46577, WO 98/06740 and WO 98/06741. However, a need remains for effective inhibitors of serine proteases such as trypsin, such as thrombin. There is a particular need for compounds, which are both orally bioavailable and selective in thrombin, which inhibits other serine proteases. Compounds that exhibit competitive inhibitory activity towards thrombin. they would be expected to be especially useful as anticoagulants and therefore in the therapeutic treatment of thrombosis and related disorders.

DESCRIPTION OF THE INVENTION According to the invention there is provided a compound of the formula I, wherein 1 represents H, CIOJR11, SiR12R13R14 or alkyl of 1 to 6 carbon atoms (in which the latter group is optionally substituted or terminated by one or more substituents selected from OR15 or (CH2) qR16); R 12, R 13 and R 14 independently represent H, phenyl or alkyl of 1 to 6 carbon atoms; R16 represents alkyl of 1 to 4 carbon atoms, phenyl, OH, C (0) OR17 or C (0) N (H) R18; R18 represents H, alkyl of 1 to 4 carbon atoms or CH2C (0) 0R19; R15 and R17 independently represent H, alkyl of 1 to 6 carbon atoms or alkylphenyl of 7 to 9 carbon atoms; R11 and R19 independently represent H or alkyl of 1 to 4 carbon atoms; and q represents 0, 1 or 2; R2 and R3 independently represent H, alkyl of 1 to 4 carbon atoms, cyclohexyl or phenyl; Rx represents a structural fragment of the formula lia, Ilb or lie, lia llb He where k, 1 and m independently represent 0, 1, 2, 3 or 4; R4 and R5 independently represent H, Si (Me) 3 / I- or 2-naphthyl, a polycyclic hydrocarbyl group, CHR41R42 or alkyl of 1 to 4 carbon atoms (in which the latter group is optionally substituted by one or more substituents halo), or cycloalkyl of 3 to 8 carbon atoms, phenyl / methylenedioxyphenyl, benzodioxanyl, benzofuranyl, dihydrobenzofuranyl, benzothiazolyl, benzoxazolyl, benzimidazolyl, coumaranonyl, coumarinyl or dihydro-coumarinyl (in which the last twelve groups are optionally substituted by one or more of alkyl of 1 to 4 carbon atoms (in which the latter group is optionally substituted by one or more substituents halo), alkoxy of 1 to 4 carbon atoms, halo, hydroxy, cyano, nitro, S02NH2, C (0) 0H or N (H) R43); R41 and R42 independently represent cyclohexyl or phenyl; R6 and R7 independently represent H, alkyl of 1 to 4 carbon atoms, cycloalkyl of 3 to 8 carbon atoms, phenyl (in which the latter group is optionally substituted by one or more of alkyl of 1 to 4 carbon atoms ( in which the latter group is optionally substituted by one or more halo substituents), alkoxy of 1 to 4 carbon atoms, halo, hydroxy, cyano, nitro, S02NH2, C (0) 0H or N (H) R44 or together with the carbon atom to which they are attached form a cycloalkyl ring of 3 to 8 carbon atoms, R43 and R44 independently represent H C (O) R 45 R 45 represents H, alkyl of 1 to 4 carbon atoms or alkoxy of 1 to 4 carbon atoms; Y represents CH2, (CH2) 2, CH = CH, (CH2) 3, CH2CH = CH or CH = CHCH2, in which the last three groups are optionally substituted by alkyl of 1 to 4 carbon atoms, methylene, oxo or hydroxy; n represents 0, 1, 2, 3 or 4; and B represents a structural fragment of the formula IVa, IVb, IVe or IVd, IVa IVb IVe IVd wherein X1, X2, X3 and X4 independently represent CH, N or N-O; X5 and X6 independently represent a single bond or CH2; one of X7, X8 and X9 represents S, O or NH, and the other two independently represent -CH =, = CH-, -N =, = N-, -N (0) = o = N (0) -; R31 represents, in all cases, one or more optional substituents selected from halo, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms or -O- (CH2) PC (O) N (R32) ( R33); p represents 0, 1, 2, 3 or 4; and R32 and R33 independently represent H, alkyl of 1 to 6 carbon atoms or cycloalkyl of 3 to 7 carbon atoms; or a pharmaceutically acceptable salt thereof. The compounds of formula I may exhibit tautomerism. All tautomeric forms and mixtures thereof are included within the scope of the invention. The compounds of formula I may also contain one or more asymmetric carbon atoms and may therefore exhibit optical isomers and / or diastereoisomerism. All diastereomers can be separated using conventional techniques, for example, chromatography or fractional crystallization. The various stereoisomers can be isolated by separation of a racemic mixture or other of the compounds using conventional techniques, by example fractional crystallization or CLAP. Alternatively, desired optical isomers can be made by reacting the appropriate, optically active starting materials under conditions that will not cause racemization or epimerization, or by derivatization, for example with a homochiral acid followed by separation of the diastereomeric derivatives by conventional means (for example CLAP, chromatography on silica). All stereoisomers are included within the scope of the invention. Allocation groups which can represent Rl p2 R 3 p. R5 R6 R7 pll pl. R13 pl4 pl5 pl6 v, -T, J, J, J, J, J, G., J? , _ \, J? , ±. , JA, R R 18 R 19, 31 R 3? R 33 and R "and with which R, R, R, R and Y can be substituted; alkoxy groups which may represent R31 and R45 and with which R4, R5, R6 and R7 may be substituted; cycloalkyl groups which may represent R2, R3, R4, R5, R6, R7, R32, R33, R41 and R42; and alkylphenyl groups which may represent R15 and R17 may be linear or branched and may be saturated or unsaturated. Alkylene groups represented by - (CH2) > .-. - (CH2)? -, - (CH2) n-, - (CH2) P- and - (CHj) q- in compounds of the formula I can be linear or branched, and can be saturated or unsaturated.

Halo groups which can represent R31, with which R4, R5, R6 and R7 can be substituted, and with which the substituents on R4, R5, R6 and R7 can be substituted, include fluorine, chlorine, bromine and iodine. In • the structural fragments of the formulas lia, Ilb and lie, the points indicated on the carbon atom that are attached to the group -C (0) - and the carbon atom, carry -OR1, R2 and R3 in a compound of Formula I (for the prevention of doubt, there is no other H atom attached to the carbon atom so indicated). The wavy line on the link in fragments IVa, IVb, IVe and IVd signify the binding position of the fragments. For the prevention of doubt, when one or. more substituents R31 is / are present, they replace one or more H atoms of CH, CH2 and / or NH groups on the appropriate rings. The skilled person will appreciate that, in the IVd structural fragment, two double bonds in the five membered ring must be present, the position of which the double bonds will depend after X7, X8 and X9 represent S, O or NH. The abbreviations are listed at the end of this specification.

Preferred compounds of the invention include those in which, when B represents a fragment structure for the formula IVa (in which X1, X2, X3 and X4 all represent CH), a structural fragment of the formula IVb or a structural fragment of the Formula IVe, R31 represents one or more substituents selected from halo, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms or -O- (CH2) P-C (0) N (RJ2) (RJJ) (ie, the substituent or substituents is not / are optional). Preferred compounds of the invention include those in which B represents a structural fragment for formula IVa. Compounds of the formula I in which the fragment is in the S configuration. The wavy lines at the nitrogen and carbon atom in the above fragment-mean the binding position of the fragment.

Preferred compounds of formula I include the compounds of Examples 1 and 2.

PREPARATION According to the invention there is also provided a process for the preparation of compounds of the formula I comprising: (a) coupling a compound of the formula V, where R1, R2, R3 and. Rx are as defined in the foregoing with a compound of formula VI, where Y, n and B are as defined in the above, for example in the presence of a system of coupling (for example oxalyl chloride in DMF, EDC, DCC, HBTU or TBTU), an appropriate base (for example pyridine, DMAP, TEA or DIPEA) and a suitable organic solvent (for example dichloromethane, acetonitrile or DMF); (b) coupling a compound of formula VII, wherein R1, R2, R3, Rx and Y are as defined in the above with a compound of the formula VIII, H2N- (CH2) n-B VIII wherein n and B are as defined above, for example in the presence of a coupling system (e.g., oxalyl chloride in DMF, EDC, DCC, HBTU or 'TBTU), an appropriate base (e.g., pyridine, DMAP) , TEA or DIPEA) and a suitable organic solvent (for example, dichloromethane, acetonitrile or DMF); (c) for compounds of formula I in which B represents a structural fragment of the formula IVa, IVb or IVe, reaction of a compound of I to formula IX, wherein B1 represents a structural fragment of the formula IVa1, IVb1 or IVe1 IVa1 IVb1 I Ve1 where Ry represents alkyl of 1 to 4 carbon atoms and R1, R2, R3, Rx, Y, n, X1, X2, X3, X4, X5, X -e6 and R, 31 are as defined above, with ammonium gas, for example at room temperature in the presence of a suitable organic solvent (for example methanol or ethanol); (d) for compounds of the formula I in which B represents a structural fragment of the formula IVa, IVb or IVe, reduction of a compound of the formula X, wherein B represents a structural fragment of the formula IVa2, IVb2 or IVe2 IVa < IVb2 IVe2 and R1, R2, R3, Rx, Y, n, X1, X2, X3, X4, X5, X6 and R, 31"are as defined above, in the presence of a suitable reducing agent (e.g. by catalytic hydrogenation in the presence of for example Pd / C or TiCl3) and an appropriate organic solvent (for example ethanol); or (e) for compounds of the formula I, wherein X 1, X 2, X 3, X 4, X 7, X 8 and / or X 9 represents NO, oxidation of a corresponding compound of the formula I in which X 1, X 2, X 3, X 4 , X7, X8 and / or X9 (as appropriate) represents N, for example under conditions that are well known to those skilled in the art. Compounds of the formula I can be prepared, in which B represents a structural fragment of the formula IVd, analogously with methods described herein or, alternatively, analogously with the methods described in the international patent applications WO 95/23609 and WO 98/06741. Compounds of formula V are commercially available, are well known in the literature or are available using known techniques. For example, compounds of formula V can be prepared by hydrolysis of a compound of formula XI, wherein R is alkyl of 1 to 6 carbon atoms or alkylphenyl of 1 to 3 carbon atoms and R1, R2, R3 and Rx are as defined above, for example at room temperature in the presence of a suitable base ( for example lithium hydroxide) and an appropriate solvent (for example THF and / or water). Compounds of formula VI can be prepared by the reaction of a compound of formula XII, wherein Y is as defined in the foregoing with a compound of formula VIII as defined above, for example under conditions such as those described above for the synthesis of compounds of formula I (process of steps ( a) and (b)). Compounds of formula VII are readily available using known techniques. For example, compounds of the formula VII can be prepared by the reaction of a compound of the formula V as defined above with a compound of the formula XII as defined above, by example under conditions such as those described in the above for the synthesis of the compounds of the formula I (process of steps (a) and (b)). Compounds of the formula IX can be prepared by known techniques. For example, compounds of the formula IX can be prepared in which B1 represents a structural fragment of the formula IVa1 or IVe1 by the reaction of a compound of the formula XIII, wherein B3 represents a structural fragment of formula IVa3 or IVe3, IVa3 IVe3 R1, R2 R- 'Rx. Y. n, X- X 'X' X4 and R31 are as defined above, with HCl (g) and an alkyl alcohol of 1 to 4 carbon atoms, for example at or below room temperature. Compounds of formula X can be prepared by known techniques. For example, compounds of the formula X can be prepared, in which B2 represents a structural fragment of the formula IVa2 or IVe2, by the reaction of a compound of the formula XIII, as defined above with HCl (g) and methanol, for example at or below room temperature, followed by reaction with hydroxylamine, or a hydrohalide salt thereof, for example at or around room temperature in the presence of an appropriate base (e.g. TEA) and a suitable solvent (eg MeOH). Compounds of the formula XI can be prepared, in which R1 and R3 represent both H, by the reduction of a compound of the formula XIV, where R, Rx and R2 are as defined above, for example at low ambient temperature (for example between -70 ° C and -5 ° C) in the presence of a suitable reducing agent (for example sodium borohydride) ) and an appropriate organic solvent (for example MeOH or EtOH). Compounds of the formula XI can be prepared, in which R1 represents H and R3 represents alkyl of 1 to 4 carbon atoms, cyclohexyl or phenyl, by the reaction of a compound of the formula XIV as defined above with a reagent organometallic of the formula XV R3 * M XV wherein R represents alkyl of 1 to 4 carbon atoms, cyclohexyl or phenyl, M represents Li or MgHal and Hal is Cl, Br or I, under conditions which are well known to those skilled in the art in the presence of an organic solvent appropriate (for example THF). It is also possible to prepare compounds of the formula XI, in which R1 represents H, by the reaction of a compound of the formula XVI, RO-C (O) -RXH XVI wherein R and Rx are as defined in the foregoing with a compound of formula XVII, R2-C (0) -R3 XVII wherein R2 and R3 are as defined in the above under conditions that are well known to those skilled in the art. Compounds of the formula XI can be prepared, in which R1, R2 and R3 all represent H, Rx, represent a structural fragment of the formula bundle, as defined above, in which neither k nor 1 represents 0, the reduction of a compound of formula XVIII, XVIII wherein Rxa represents a structural fragment of the formula lia, as defined above, in which neither k nor 1 represents 0, and R is as defined above, in% the presence of a reducing agent suitable (for example borane) in the presence of an appropriate organic solvent (for example THF). Compounds of formula XIII can be prepared by coupling a compound of formula VII as defined above for a compound of formula XIX, H2N- (CH2) a-I_r XIX wherein n and B are as defined in the foregoing, for example under conditions such as those described above for the synthesis of compounds of formula I (process of steps (a) and (b)). Compounds of the formula XIV are either known from, or may be prepared analogously to, the methods described in J. Org. Chem., 54, 3831 (1989). Compounds of formula XVIII are well known in the literature or can be prepared using known techniques, for example by the reaction of a suitable malonic acid derivative with an alkylating agent of formula XX, R? L XX wherein L is a leaving group (for example halo (Cl, Br, I) or tosyl) and Rxa is as defined above, for example in the presence of a suitable base (e.g., sodium hydride or ethoxide). sodium) and an appropriate organic solvent. Compounds of the formula VIII, XII, XV, XVI, XVII, XIX and XX and derivatives thereof, are either commercially available, are known in the literature, or can be obtained either by analogy with the process described herein , or by conventional synthetic procedures, according to standard techniques, from readily available starting materials using appropriate reagents and reaction conditions. Substituents in phenyl groups can be inter-converted into the contents of compounds of formulas I, V, VI, VII, VIII, IX, X, XI, XIII, XIV, XVI, XVIII, XIX and XX, using standard techniques . Compounds of the formula I can be isolated from their reaction mixtures using conventional techniques. It will be appreciated by those skilled in the art that in the process described in the foregoing, the Functional groups of intermediate compounds may need to be protected by protecting groups. Functional groups that are desirable to protect include hydroxy, amino and carboxylic acid. Suitable protecting groups for hydroxy include trialkylsilyl or diarylalkysilyl groups (for example t-butyldimethylsilyl, t-butyldiphenylsilyl or trimethylsilyl) and tetrahydropyranyl. Suitable protecting groups for the carboxylic acid include alkyl of 1 to 6 carbon atoms or benzyl esters. Suitable protecting groups for amino, amidino and guanidino include t-butyloxycarbonyl or benzyloxycarbonyl. Amidino and guanidino nitrogens can be either mono- or diprotected. The protection and deprotection of functional groups can be carried out before or after coupling. For example, compounds of formula I can be prepared by processes comprising the coupling of an N-acylated amino acid or an N-protected amino acid. When an N-protected amino acid is used, the acyl group can be added after coupling and deprotection of the nitrogen atom can then be carried out using standard methods after this.

Protective groups can be removed according to techniques that are well known to those skilled in the art and as described hereinafter. The use of protecting groups is fully described in "Protective Groups in Organic Chemistry", edited by JW F McOmie, Plenum Press (1973), and "Protective Groups in Organic Synthesis", 2nd edition, T W Greene & P G M Wutz, Wiley-Interscience (1991). Certain protected derivatives of compounds of the formula I, which can be made prior to a final deprotection step to form compounds of the formula I, are novel. According to another aspect of the invention there is provided a compound of the formula la, where Ba represents a structural fragment of the formula IVe, IVf, IVg or IVh IVe IVf IVg IVh wherein D1 and D2 independently represent, in each case, H, OH, ORa, OC (0) Rb, OC (0) ORc, C (0) ORd, C (0) Re, and Ra, Rb, Rc, Rd and Rp independently represent alkyl of 1 to 12 carbon atoms (in which the latter group is optionally interrupted by oxygen and / or substituted by halo) ', phenyl, naphthyl, alkylphenyl of 1 to 3 carbon atoms (in which three groups are optionally substituted by alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms, nitro or halo) or - (C ( Rf) (Rg)) 20C (O) C (Rh), Rf, Rg and Rh independently represent H or alkyl of 1 to 4 carbon atoms, and R1, R2, R3, Rx, Y, n, X1, X2, X3, X4, X5, X6, X7, X8, X9 and R31 are as defined above, or a pharmaceutically acceptable salt thereof, with the proviso that D1 and D2 both do not represent H.

Alkyl groups which can represent Ra, Rb, Rc, Rd, Re, Rf, Rg and Rh, and with which Ra, Rb, Rc, Rd and Re can be substituted, can be linear or branched, can be saturated or unsaturated , and they can be cyclic, acyclic or cyclical / acyclic. Alkoxy groups with which Ra, Rb, Rc, Rd and Re can be substituted, can be linear or branched, can be saturated or unsaturated, and can be cyclic, acyclic or cyclic / acyclic. The alkyl part of alkylphenyl groups which may represent Ra, Rb, Rc, Rd and Re, may be linear or branched, and may be saturated or unsaturated. Halo groups with which Ra, Rb, Rc and Re can be substituted include fluorine, chlorine, bromine and iodine. The wavy line on the link in the fragments IVe, IVf, IVg and IVh, means the binding position of the fragments. Preferred compounds of the formula include those wherein D2 represents H and D1 represents OH, 0CH3, OC (0) Rb or C (0) 0Rd, wherein Rb and Rd are as defined above. Structural preferences mentioned above are also applied for compounds of the formula I to compounds of the formula la.

Compounds of the formula la can also be prepared directly from the compounds of the formula I according to techniques well known to those skilled in the art. For example, compounds of the formula la can be prepared in which Ba represents a structural fragment of the formula IVe, IVf or IVg, and in which D1 or D2 represent OH, as described above for compounds of the formula X, or by analogous methods. Alternatively, compounds of the formula la, in which Ba represents a structural fragment of the formula IVe, IVf or IVg, and in which D1 or D2 represent OH or 0Ra, wherein Ra is as defined above, can be prepare from compounds of formula XIII as defined above by the reaction with a compound of formula XXI, H2NORal XXI wherein Ral represents H or Ra and Ra is as defined above, for example at 40 to 60 ° C, in the presence of a suitable base (for example, TEA) and an appropriate organic solvent (for example, THF) , CH3CN, DMF or DMSO).

Alternatively, compounds of the formula can be prepared via other protected derivatives of the formula according to techniques well known to those skilled in the art. For example, compounds of the formula la can be prepared in which D1 or D2 represent 0C (0) 0Rc, and Rc is as defined above, by the reaction of a corresponding compound of the formula la in which D1 or D2 (as appropriate) represents OH with a compound of the formula XXII, R ° C. { 0) -0-C (0) Re XXII wherein R c is as defined above, for example at room temperature in the presence of a suitable base (e.g., TEA, pyridine or DMAP) and an appropriate organic solvent. In addition, compounds of the formula la can be prepared in which D1 or D2 represent OH, by the reaction of a corresponding compound of the formula la in which D1 or D2 (as appropriate) represents C00Rd and Rd is as defined above with hydroxylamine (or a hydrohalide salt thereof), for example at 40 ° C in the presence of a suitable base (for example TEA) and an appropriate organic solvent (for example THF).

Compounds of formulas XXI and XXII are commercially available, are well known in the literature, or are available using known techniques. It will also be appreciated by those skilled in the art that, although such protected derivatives of compounds of the formula I (e.g.

, Formula 1) can not possess pharmacological activity as such, they can be administered parenterally or orally and thereafter metabolized in the body to form compounds of the invention which are pharmacologically active. Such derivatives can therefore be described as "prodrugs". All prodrugs of compounds of the formula la are included within the scope of the invention. Protected derivatives of compounds of the formula I which are particularly useful as prodrugs include compounds of the formula la. further, certain compounds of the formula I can act as prodrugs of other compounds of the formula I. Compounds of the formula I, pharmaceutically acceptable salts, tautomers and tereoisomers thereof, as well as prodrugs thereof (including compounds of the The formula which are prodrugs of compounds of the formula I) are hereinafter referred to as "the compounds of the invention". The person skilled in the art will appreciate that, in order to obtain compounds of the invention in an alternative, and, on some occasions, more conveniently, the steps of the individual process mentioned herein may be performed in a different order, and / or the individual reactions can be carried out at a different stage in the total route (that is, substituents a and / or chemical transformations carried out on, intermediates different from those associated in the foregoing can be added with a particular reaction). This will depend on the factors such as the nature of different functional groups. present in a particular substrate, the availability of key intermediaries and the strategy of the protective group (if any) that is adopted. Clearly, the type of chemistry involved will influence the choice of reagent that is used in the synthetic steps, the need, and type, of protective groups that are employed, and the sequence to perform the synthesis.

MEDICAL AND PHARMACEUTICAL USE The compounds of the invention are useful because they possess pharmacological activity. They are therefore indicated as pharmaceuticals. According to another aspect of the invention of this. The compounds of the invention are provided for use as pharmaceuticals. In particular, the compounds of the invention are potent inhibitors of proteases such as trypsin, especially thrombin, either as such or, in the case of prodrugs, after administration to mammals including man, for example as demonstrated in the tests described. later. The compounds of the invention are thus expected to be useful in those conditions where inhibition of thrombin is required. The compounds of the invention are thus indicated in the treatment or prophylaxis of thrombosis and hypercoagulability in the blood and tissues of animals including man. It is known that hypercoagulability can lead to thrombo-embolic diseases. Conditions associated with hypercoagulability and diseases Thrombo-embolisms that may be mentioned include resistance to activated protein C, such as the V mutation factor (Leiden factor V), and hereditary or acquired deficiencies in antithrombin III, protein C, protein S, heparin cofactor II. Other known conditions that are associated with hypercoagulability and thromboembolic disease include circulating antiphospholipid antibodies (anticoagulant lupus), homocysteinemia, heparin-induced thrombocytopenia, and defects in fibrinolysis. The compounds of the invention are thus indicated both in the therapeutic and prophylactic treatment of these conditions. The compounds of the invention are further indicated in the treatment of conditions where there is an undesirable excess. of thrombin without signs of hypercoagulability, for example in neurodegenerative diseases such as Alzheimer's disease. Particular disease states that may be mentioned include the therapeutic and / or prophylactic treatment of venous thrombosis and pulmonary embolism, arterial thrombosis (for example in myocardial infarction or heart failure, unstable angina, attack or shock based on thrombosis and peripheral arterial thrombosis) and systematic embolism that usually originates from the atrium during arterial fibrillation or from the left ventricle after infarction to the transmural myocardium. In addition, the compounds of the invention are expected to be useful in the prophylaxis of reocclusion (ie, thrombosis) after thrombolysis, percutaneous transluminal angioplasty (PTA) and coronary bypass operations; the prevention of re-thrombosis after microsurgery and vascular surgery in general. Other indications include the therapeutic and / or prophylactic treatment of disseminated intravascular coagulation caused by bacteria, multiple trauma, intoxication or any other mechanism; anticoagulant treatment when the blood is in contact with external surfaces in the body such as vascular grafts, vascular stents, vascular catheters, mechanical and biological prosthetic valves or any other medical device; and anticoagulant treatment when the blood is in contact with medical devices outside the body such as during cardiovascular surgery using a heart-lung machine or in hemodialysis.

In addition, for its effects on the coagulation process, thrombin is known to activate a large number of cells (such as neutrophilis, fibroblasts, endothelial cells and smooth muscle cells). Therefore, the compounds of the invention may also be useful for the therapeutic and / or prophylactic treat of distress syndrome or idiopathic and adult respiratory distress, pulmonary fibrosis following treat with radiation or chemotherapy, septic shock, septicemia, inflammatory responses , which include, but are not limited to, edema, acute or chronic atherosclerosis such as coronary artery disease, cerebral arterial disease, peripheral artery disease, reperfusion injury, and restenosis after percutaneous trans-luminal angioplasty (PTA). Compounds of the invention that inhibit trypsin and / or thrombin in the treat of pancreatitis may also be useful. According to another aspect of the present invention, there is provided a method for the treat of a condition, wherein inhibition of thrombin is required or desired, which method comprises administering a therapeutically effective amount of a compound of the invention. invention, or a pharmaceutically acceptable salt thereof, for a person suffering from, or who is susceptible to, such a condition. The compounds of the invention will normally be administered orally, intravenously, subcutaneously, buccally, rectally, dermally, nasally, tracheally, bronchially, or by any other parenteral route or inhalation route, in the form of pharmaceutical preparations comprising an active compound either as a free base, or a pharmaceutically acceptable, non-toxic, organic or inorganic acid addition salt, in a pharmaceutically acceptable dosage form. Depending on the disorder and the patient to be treated and the route of administration, the compositions may be administered in various doses. The compounds of the invention can also be combined and / or co-administered with any antithrombotic agent with a different mechanism of action, such as antiplatelet agents of acetylsalicylic acid, ticlopidine, clopidogrel, thromboxane receptor and / or synthetase inhibitors, antagonists of the fibrinogen receptor, prostacyclin mimetic inhibitors and phosphodiesterase and ADP receptor antagonists (P2T).

The compounds of the invention can also be combined and / or co-administered with thrombolytics such as tissue plasminogen activator (natural, recombinant or modified), streptokinase, urokinase, prourokinase, anisoylated plasminogen-streptokinase activator complex (APSAC), activators of the plasminogen of the animal salivary gland, and the like, in the treat of thrombotic diseases, in particular myocardial infarction. In accordance with another aspect of the invention this way a pharmaceutical formulation is provided which includes a compound of the invention, in admixture with a pharmaceutically acceptable adjuvant, diluent or carrier. Suitable daily doses of the compounds of the invention in therapeutic treat of humans are approximately 0.001-100 mg / kg of body weight in peroral administration and 0.001-50 mg / kg of body weight in parenteral administration. The compounds of the invention have the advantage that they can be more effective, be less toxic, be more long acting, have a broader range of activity, be more potent, produce fewer side effects, be more easily absorbed than, or that can have other properties useful pharmacological agents, compounds known in the prior art.

BIOLOGICAL TESTS Test A Determination of Thrombin Coagulation Time (TT) Human thrombin was incubated (T 6769, Sigma Chem. Co. ) in buffered solution, pH 7.4, 100 μl and inhibitor solution, 100 μl, for one minute. Accumulated normal cipher human plasma, 100 μl, and coagulation time measurement in an automatic device (KC 10, Amelumg) were then added. The coagulation time in seconds was plotted against the inhibitory concentration, and the IC50TT by interpolation. The IC5_TT is the concentration of the inhibitor in the test that doubles the coagulation time of thrombin for human plasma.

Test B Determination of Thrombin Inhibition with a Robotic, Chromogenic Assay The potency of the thrombin inhibitor was measured with a chromogenic substrate method, in a Robotic microplate processor Plato 3300 (Rosys AG, CH-8634 Ho brechtikon, S i tzerland), using 96-well medium volume microtiter plates (Costar, Cambridge, MA, USA, Cat. No. 3690). Stock solutions of test substance were diluted in DMSO (72 μl, 1 mmol / 1) in a 1: 3 series (24 + 48 μl) with DMSO to obtain ten different concentrations, which were analyzed as samples in the assay. 2 μl of the test sample was diluted with 124 μl of the assay buffer, 12 μl of chromogenic substrate solution (S-2366, Chromogenix, Molndal, Sden) in assay buffer and finally 12 μl of a -trombine, (Human a-thrombin, Sigma Chem. Co.) both in assay buffer, and the samples are mixed. The final test concentrations were: test substance 0.00068 - 13.3 umol / 1, S-2366 0.30 mmol / 1, a-thrombin 0.020 NIHU / ml. The increase in linear absorbance during 40 minutes of incubation at 37 ° C was used to calculate the percentage inhibition for the test samples, compared to blank without inhibitor. The IC50-robotic valve, which corresponds to the concentration of the inhibitor that causes 50% inhibition of thrombin activity, was calculated from a log dose vs. % of the inhibition curve.

Test C Determination of the K Inhibition Constant for the Human Thrombin Id determinations were made using a chromogenic substrate method, performed at 37 ° C on a Cobas Bio centrifugal analyzer (Roche, Basel, Switzerland). Residual enzyme activity was determined after incubation of human a-thrombin with various concentrations of the test compound at three different substrate concentrations, and was measured as the change in optical absorbance at 405 nm. Solutions of the test compound (100 μl, usually in buffer or saline containing 10 g / 1 BSA) were mixed with 200 μl of human a-thrombin (Sigma Chem. Co.) In assay buffer (0.05 mol / 1 Tris). -HCl pH 7.4, 0.15 ionic strength adjusted with NaCl) containing BSA (10 g / 1), and analyzed as samples in the Cobas Bio. A 60 μl sample, together with 20 μl of water, was added to 320 μl of substrate S-2238 (Chromogenix AB, Molndal, Sden) in assay buffer, and the absorbance change (ÅA / min) was monitored. The final concentrations of S-2238 were 16, 24 and 50 μmol / l and 0.125 NIH U / ml of thrombin.

The steady-state reaction rate was used to construct Dixon traces, i.e., inhibitor concentration diagrams vs l / (? A / min). For competitive, reversible inhibitors, data points for different substrate concentrations typically form straight lines that intercept Test D Determination of Activated Partial Thromboplastin Time (APTT) APTT was determined in normal human citrated plasma accumulated with the PTT reagent Auto ated 5 manufactured by Stago. The inhibitors were added to the plasma (10 μl of inhibitor solution to 90 μl of plasma) followed by the reagent and calcium chloride solution and APTT were determined in the mixture by using the KC10 coagulation analyzer (Amelumg) according to the instructions from the reactive producer. The coagulation time in seconds is plotted against the inhibitory concentration in plasma and the IC5.APTT was determined by interpolation. IC50APTT was defined as the concentration of the inhibitor in human plasma that doubles the Activated Partial Thromboplastin Time.

Test E Determination of Thrombin Time ex vi Vo inhibition of thrombin after oral or parenteral administration of the compounds of formula I and, dissolve in ethanol: SolutolMR: water (5: 5: 90), were examined in conscious rats which, one or two days before the experiment, were equipped with a catheter for blood sample of the carotid artery. Blood samples on the day of the experiment were separated at fixed times after administration of the compound in plastic tubes containing 1 part sodium citrate solution (0.13 moles per 1) and 9 parts blood. The tubes were centrifuged to obtain poor quality plasma platelets. Plasma was used for determination of thrombin time as described below. Crude rat plasma was diluted, 25 μl with a saline solution, 0.9%, 25 μl, and plasma coagulation was initiated by the addition of human thrombin (T 6769, Sigma, Chem. Co., USA) in a buffer solution. , pH 7.4, 25 μl. The coagulation time was measured in an automatic device (KC lOA-micro, Amelumg, Germany).

When a compound of the formula la was administered, concentrations of the appropriate active thrombin inhibitor of the formula I are estimated in rat plasma, by the use of standard curves, related to the thrombin time in the coded rat plasma, accumulated at known concentrations of the corresponding "active" thrombin inhibitor, dissolved in saline. Based on the estimated plasma concentrations of the active plasma inhibitor of formula I (assuming that the prolongation of the thrombin time is caused by the compound mentioned above) in the rat, the area under the plasma concentration curve-time after of the oral and / or parenteral administration of the corresponding compound of. The formula was calculated (AUCpd) using the trapezoidal rule and extrapolation of data to infinity. The bioavailability of the active thrombin inhibitor of the formula I after oral or parenteral administration of the compound of the formula la was calculated as follows: [(AUCpd / dose) / (AUCactive, parenteral / dose] x 100 where AUCactive, parenteral represents the AUC obtained after parenteral administration of the active thrombin inhibitor corresponding to formula I to conscious rats as described above.

Test F Determination of Thrombin Time in Extinct Urine The amount of the active thrombin inhibitor of formula I was excreted in urine after oral or parenteral administration of the compounds of the invention, dissolved in ethanol: Solutol ™: water (5: 5: 90), it was estimated by determining the thrombin time in ex vivo urine (assuming that the prolongation of thrombin time is caused by the compound mentioned above). They were placed, conscious rats in metabolism cages, allowing to separate the collection of urine and feces, for 24 hours followed by the oral administration of compounds of the invention. The thrombin time was determined in the urine collected as described below. Accumulated normal cipher human plasma (100 μl) was incubated with the concentrated rat urine, or saline dilutions thereof, for one minute. Then the plasma coagulation was started by the administration of human thrombin (T 6769, Sigma Chem. Co.) in buffer solution (pH 7.4, 100 μl). The coagulation time was measured in an automatic device (KC 10; Amelumg). The concentrations of the active thrombin inhibitor of formula I in rat urine were estimated by the use of standard curves that relate to the thrombin time in accumulated normal cipher human plasma at known concentrations of the active thrombin inhibitor mentioned above. , dissolved in concentrated rat urine (or saline dilutions thereof). By multiplying the total rat urine production during the 24 hour period with the estimated average concentration of the active inhibitor mentioned in the above in the urine, the amount of the active inhibitor excreted in the urine (AMOUNTpd) could be calculated. The bioavailability of the active tro-bin inhibitor of formula I after oral or parenteral administration of the prodrug was calculated as follows: [(AMOUNTpd / dose) / (MOUNTactive, parenteral / dose] x 100 wherein AMOUNTactive, parenteral represents the amount excreted in the urine after parenteral administration of the corresponding active thrombin inhibitor of formula I to conscious rats as described above. The invention is illustrated by the form of the following examples.

EXAMPLES General Experimental Procedures The mass spectrum was recorded on a Finnigan MAT TSQ 700 triple quadrupole mass spectrometer equipped with an electrovaporizing interface zone (FAB-EM) and a VG Platform II mass spectrometer equipped with an electrovaporizing interface (LC-). EM). Measurements of 13E NMR and 13C NMR were made on BRUKER ACP 300 and Varian UNITY plus 400, 500 and 600 spectrometers, operating at frequencies of 1E of 300.13, 399.96, 499.82 and 599.94 MHz respectively, and at frequencies of 13C of 75.46, 100. 58, 125.69 and 150.88 MHz respectively. Preparative CLAP was performed on reverse phase columns (250 mm, 20 or 50 mm, Chromasil C8 phase from 5 to 7 μM) with flow rates of 10 to 50 ml / minute using a UV detector (240 to 290 nm).

Example 1 (2,5-diMeO) Ph- (5) - or (R) -CH (CH2OH) -CO-Pro-Pab (2-Cl) x HOAc (i) 4-Azidomethyl-3-chlorobenzoni trile A mixture of 8.0 g is stirred vigorously (0.035 moles) of 4-bromomethyl-3-chlorobenzonitrile (J. Pharm, Sci., (1986) 75, 410), 2.7 g (0.042 moles) of sodium azide, 1.2 g (3.4 mmoles) of tetrabutylammonium acid sulfate , 0.30 g (3.4 mmoles) of sodium hydrogencarbonate, 7 ml of water and 20 ml of toluene for 3 days. The phases are separated and the aqueous layer is extracted three times with ether. The combined organic phase is washed with water, dried over sodium sulfate and evaporated to give 6.7 g (100%) of the subtitle compound. NMR-aH (300 MHz, CDC13): d 4.60 (s, 2H), 7.57 (d, 1H), 7.61 (m, 1H), 7.70 (d, 1H) (i i) 4-Aminomethyl-3-chlorobenzoni trile Dissolve 4-azidomethyl-3-chlorobenzoni trile (1.0 g, 5.2 mmol, from step (i) above), in 9 ml of water and 1 ml of ethanol. Triphenylphosphine (1.5 g) is added, and the mixture is stirred overnight. The ethanol is evaporated and the residue divided between 1M HCl and benzene. The aqueous layer is extracted several times with Benzene and then lyophilized. The yield of amine hydrochloride was 0.54 g (51%). NMR-aH (400 MHz; D20) HCl salt; 6 4.42 (s, 2H), 7.69 (d, 1H), 7.81 (dd, 1H), 7.98 (d, 1H) (iii) Ethyl ester of (R, S) -3-hydroxy-2- (2,5-dimethoxy phenyl) propionic acid It is added to a solution of 3-OXO-2- (2,5-dimethoxyphenyl) propionic acid ethyl ester (7.6 g, 30 mmol, prepared according to the method described in J. Org Chem. 54, 3831 (1989)) in ethanol NaBH 4 (2 equivalents) at -5 ° C. After stirring for 2 hours at -15 ° C, and 4 hours at -5 ° C, water is added, the reaction mixture is concentrated and the resulting mixture is extracted with ethyl acetate. The organic phase is washed with brine, dried (MgSO 4) and concentrated to give the sub-title compound. Yield of 7.7 g (100%). (iv) (2, 5-diMeQ) Ph-. { R, S) CH (CH2QH) -COOH Ethyl ester is dissolved from the stage (iii) above (7.4 g, 29 mmol) in THF: water (1: 1).

LiOH x H20 (2 equivalents) is added and the reaction mixture is stirred for 2 hours at room temperature.

Concentrate and extract the reaction mixture with CH2C12. The aqueous phase (pH 2) is acidified with HCl (2M) and extract three times with CH2C12. The aqueous phases are combined, dried (NaS0) and concentrated to give the subtitle compound. Yield of 5.4 g (82%). NMR-aH (400 MHz, CDC13): d 6.89 (d, 1H), 6.85 (d, 1H); 6.78 (dd, 1H); 4.86 (broad, 2H); 4.14 (dd, 1H); 3.98 (dd, 1H); 3.78 (s, 3H); 3.72 (s, 3H); 3.67 (dd, 1H) (v) (2, 5-diMeO) Ph- (R, S) CH (CH2OH) -CQ-Pro-OBn. 1.8 ml (9.6 mmol) of diisopropyl-ethylamine is added to an ice-cooled mixture of 0.50 g (2.2 mmoles) of (2, 5-diMeO) Ph- (R, S) CH (CH 2 OH) -COOH (from step (iv) above), 0.58 g (2.4 mmoles) of H-Pro-OBn and 0.77 g (2.4 mmol) of TBTU in 10 ml of DMF. The mixture is stirred at room temperature overnight, and then poured into 1M HCl and extracted twice with ethyl acetate. ethyl: toluene (1: 1). The combined organic layer is washed with NaHCO (aqueous) and water, dried (Na 2 SO 4) and evaporated. The product was pure and is used directly in the next stage. Yield: 0.91 g (100%). NMR-JH (500 MHz; CDC13) diastereomeric mixture: d 1.8-2.3 (m, 4H), 3.0-3.1 (, 1H), 3.18 (m, 1H, diastereomer), 3.34 (m, 1H, diastereomer), 3.46 ( m, 1H, diastereomer), 3.65-3.7 (m, 1H), 3.75-3.9 (m, 7H, 4 singles thereof to 3.77, 3.80, 3.85 and 3.87 ppm), 3.95-4.05 (m, 1H), 4.39 (m, 1H, diastereomer), 4.46 (m, 1H, diastereomer), 4.58 (m, 1H, diastereomer), 4.63 (m, 1H, diastereomer), 5.18 (d, 1H, diastereomer), 5.26 (s, 1H, diastereomer), 5.32 (d, 1H), diastereomer), 5.37 (s, 1H, diastereomer), 6.8-7.0 (m, 3H), 7.2-7.5 (m, 5H) (vi) (2, 5-diMeO) Ph- (R, S) CH (CH2OH) -CO-Pro-OH Hydrogenated (2, 5-diMeO) Ph- (R, S) CH (CH2OH) -CO- Pro-OBn (0.91 g; 2.2 mmole; from step (v) above) for 2 hours at atmospheric pressure over 50 mg of 10% Pd / C in 25 ml of ethanol. The mixture is filtered through celite and evaporated to give 0.71 g. (100%) of the subtitle compound .. NMR-XH (500 MHz; CDC13) mixed tereomeric dias: d 1.8- 2.3 (m, 4H), 3.02 (m, 1H, diastereomer), 3.15 (m, 1H, diastereomer ), 3.47 (m, 1H, diastereomer), 3.7-3.8 (m, 5H, of the same the other diastereomer corresponding to 3.47, and two singlets at 3.74 and 3.76 ppm), 3.83-3.85 (m, 3H), 4.0-4.1 (m, 1H), 4.38 (m, 1H, diastereomer) , 4. 50 (m, 1H, diastereomer), 4.56 (m, 1H, diastereomer), 4. 63 (m, 1H, diastereomer), 6.8-6.9 (m, 3H) (vii) (2, 5-diMeO) Ph- (R and 5) CH (CH2OH) -CO-Pro-NHCH2-Ph (2-C1,4-CN) 0.76 ml (4.4 mmol) of diisopropylethylamine is added to an ice-cooled mixture of 0.35 g (1.1 mmol) of 2, 5-diMeO) Ph- (R, 5) CH (CH 2 OH) -CO-Pro-OH (from from step (vi) above), 0.22 g (1.1 mmol) of 4-aminomethyl-3-chlorobenzonitrile (from step (ii) above) and 0.35 g (1.1 mmol) of TBTU in 10 ml of DMF. The mixture is stirred at room temperature for two days, poured into 1M HCl and extracted twice with ethyl acetate: toluene (1: 1). The combined organic layer is washed with NaHCO 3 (aqueous) and water, dried (Na 2 SO 4) and evaporated. The crude product is subjected to flash chromatography on silica gel with EtOAc: MeOH (99: 1) as eluent. The diastereomers are separated. Total yield: 0.48 g (94%). The initial diastereomer (A) is isolated in a yield of 0.22 g. Diastereomer A NMR- '? (400 MHz, CDC13): d 1.7-2.1 (m, 3H), 2.34 (m, 1H); 2.90 (m, 1H), 3.12 (m, 1H), 3.61 (m, 1H), 3.73-3.8 (, 4H, a singlet thereof to 3.75), 3.82 (s, 3H), 4.01 (m, 1H), 4.44 (m, 1H), 4.56 (m, 1H), 4.65 (, 1H), 6.75-6.85 (m, 3H), 7.53 (d, 1H), 7.56 (dd, 1H), 7.65 (d, 1H), 7.68 (m, 1H) (viii) (2, 5-diMeO) Ph- (i.5) CH (CH2OH) -CO-Pro-NHCH2- Ph (2-Cl, 4-C (NH) OMe) Dissolves (2, 5-diMeO ) Ph- (R or S) CH (CH20H) -CO-Pro-NHCH2-Ph (2-Cl, 4-CN) (0.22 g, 0.47 mmol, diastereomer A from step (vii) above) in 25 ml of methanol saturated with hydrogen chloride gas and place it in the refrigerator for two days. The evaporation produces an unpurified material that is used in the next step without further purification. MS (M + 1) + 504 (ix) (2,5-diMeO) Ph- (i. or 5) CH (CH2OH) -CO-Pro-Pab (2-Cl) x HOAc Half is dissolved. of the unpurified material from step (viii) above in 25 ml of methanol saturated with ammonia and allowed to stand at room temperature for five days. The solvent is evaporated and the residue is purified in preparative CLAP with CH3CN: 0.1 M NH4OAc (30:70). Water lyophilization gave 9 mg (7%) of the desired product. NMR-aH (400 MHz; D20) rotamers: d 1.8-2.4 (m, 7H, singlet thereof at 1.93 ppm), 3.35 (m, 1H), 3.63 (, 1H), 3.7-3.9 (m, 8H, singlettes of the same at 3.80, 3.81 and 3.86 ppm (rotamers)), 4.05 (m, 1H), 4.4-4.65 (, 3H), 6.82 (d, 1H, major rotamer), 6.86 (d, 1H, minor rotamer) 6.95-7.05 m. 1H) 7.0¡ (, 1H, major rotamer), 7.23 (d, 1H, minor rotamer), 7.59 (d, 1H), 7.63 (dd, 1H, minor rotamer), 7.73 (dd, 1H, major rotamer), 7.82 (d, 1H, minor rotamer), 7.98 (d, 1H, major rotamer) NMR-13.C (100 MHz, D20) carbonyl carbons and amidine: d 176.2, 174.6, 166.7 Example 2 (2, 5-diMeO) Ph- (i 5) CH (CH 2 OH) -CO-Pro-Pab (2-Me) x HOAc (i) 3-Methyl-4-vinylbenzoni trile 0.75 g (0.65 mmoles) of tetrakis (triphenylphosphine) palladium (0) is added to a solution of 5.1 g (0.026 moles) of 4-bromo-3-methylbenzonitrile and 8.3 g (0.026 moles) of vinyl tributyltin in 250 ml of toluene under argon, and the reaction is heated to reflux overnight. The mixture is filtered through a pad of celite and evaporated. The residue is chromatographed on silica gel with heptane: EtOAc (1: 1) as eluent. Yield: 4.0 g (100%). NMR-H (300 MHz, CDC13): d 2.36 (s, 3H), 5.46 (d, 1H), 5.73 (d, 1H), 6.90 (dd, 1H), 7.4-7.5 (, 2H), 7.52 (d , 1 HOUR) (ii) 4-Hydroxymethyl-3-methylbenzonitrile 3-Methyl-4-vinylbenzonitrile (0.40 g, 2.8 mmol, from step (i) above) is dissolved in 50 ml of methanol and cooled to -70 ° C. They are bubbled through ozone (2 equivalents), and then 0.20 g are added (5.3 mmoles) of sodium borohydride and 5 ml of water and the cooling bath is removed. After 4 hours the methanol is evaporated and the residue is partitioned between 1M HCl and ether. The aqueous layer is extracted twice with ether and the combined organic phase is washed with water, dried (Na2SO4) and evaporated to yield 0.37 g (90%) of the desired product, which is used in the next step without further purification. NMR ^ H (400 MHz, CDCl 3): d 2.29 (s, 3H), 3.07 (broad, 1H), 4.69 (s, 2H), 7.37 (s, 1H), 7.45 (d, 1H), 7.53 (d, 1H) (iii) 4-Methanesulfonyloxy-3-methylbenzonitrile 1.2 g (0.010 mole) of me tyl-sulphonyl chloride is added dropwise to a solution of 1.5 g (0.010 moles) of 4-hydroxymethyl-3-methylbenzonitrile (from step (ii) above) and 1.0 g (0.010 moles) of triethylamine in 50 ml of methylene chloride at 0 ° C. The reaction mixture is stirred at room temperature for 3 hours, it is washed with 1M HCl, water, dried (Na2SO4) and evaporated. Yield: 2.1 g (91%). NMR-1! - (300 MHz; CDC13): d 2.41 (s, 3H), 3.03 (s, 3H), . 28 (s, 2H), 7.4-7.6 (m, 3H) (iv) 4-Azidomethyl-3-methylbenzonitrile. 1.0 g (0.015 mole) of sodium azide and 10 ml of water are added to a solution of 2.1 g (9.3 mmol) of 4-methanesulfonyloxy-3-methylbenzonitrile (from step (iii) above) in 20 ml of DMF. The reaction mixture is stirred for 1.5 hours at room temperature, poured into 200 ml of water and extracted three times with ether. The combined organic phase is washed several times with water, dried (Na 2 SO 4) and evaporated. Yield: 1.4 g (87%). NMR-aH (300 MHz, CDC13): d 2.39 (s, 3H), 4.40 (s, 2H), 7. 40 (d, 1H), 7.45-7.55 (m, 2H) (v) 4-Aminomethyl-3-methylbenzoyl itryl The subtitle compound is prepared according to the method described in Example 1 (ii) above of 1.4 g (8.1 mmol) of 4-azidomethyl-3-methylbenzonitrile (from step (iv) above) and 2.3 g (9.0 mmol) of triphenylphosphine in 18 ml of ethanol and 2 ml of water. Yield: 0.60 g (contaminated by triphenylphosphine oxide, estimated current yield: 0.36 g (28%)). RMN-1 !. (300 MHz, CDC13): d 2.38 (s, 3H), 3.90 (s broad, 2H), 7.3-7.4 (m, 3H) darkened by triphenylphosphine oxide (vi) (2, 5-diMeO) Ph- (R and 5) CH (CH20H) -CO-Pro-NHCH2-P (2-Me, 4-CN) The subtitle compound is prepared according to the method described in Example l (vii) above of 0.37 g (1.2 mmol) of (2, 5-diMeO) Ph- (R, S) CH (CH2OH) -CO-Pro-OH (see Example l (vi) above) , 0. 42 g of 4-aminomethyl-3-methybenzoni tryl (from step (v) above, estimated content of pure material: 0.25 g (1.7 mmol)), 0.38 g (1.2 mmol) of TBTU and 0.62 g ( 4.8 mmol) of diisopropylethylamine in 10 ml of DMF. The crude product is subjected to flash chromatography on silica gel with EtOAc: acetone (9: 1) as eluent. The two diastereomers are separated. Total yield: 0.58 g contaminated by triphenylphosphine oxide; Current estimated yield: 0.45 g (87%). The first diastereomer which is eluted as diastereomer A. Diastereomer A is isolated (yield: 0.30 g, contaminated by triphenylphosphine oxide, estimated current yield: 0.20 g) RMN-1 !! (600 MHz, CDC13): d 1.7-2.1 (m, 3H), 2.3-2.35 (m, 4H, a singlet thereof at 2.32 ppm), 3.1-3.2 (, 1H), 3.65 (m, 1H), 3.7 -3.8 (, 4H, a singlet thereof at 3.73 ppm), 3.80 (s, 3H), 3.98 (, iH), 4.4-4.5 (m, 3H), 4.65 (m, 1H), 6.7-6.9 (m, 3H), 7.37 (d, 1H), 7. 41 (s, 1H), the remaining aromatic signal obscured by triphenylphosphine oxide (vii) (2, 5-diMeO) Ph- (R or S) CH (CH2OH) -CQ-Pro-NHCH2-Ph (2-Me, -C (NH) OMe) Dissolves (2, 5-diMeO) Ph- (R or S) CH (CH2OH) -CO-Pro-NHCH2-Ph (2-Me, 4-CN) (0.30 g, estimated content of pure material: 0.20 g (0.44 mmol), diastereomer A from step (vi) above) in 25 ml of methanol saturated with hydrogen chloride gas and allowed to stand at room temperature overnight. The evaporation produces an unpurified material that is used without further purification. According to CCF (EtOAc: acetone; 9: 1) the starting material is not left. (viii) (2, 5-diMeO) Ph- (R or S) CH (CH2OH) -CO-Pro-Pab (2-Me) (OH) The unpurified product is dissolved from step (vii) above in 10 ml of methanol and 0. 14 g (2.0 mmol) of hydroxylamine hydrochloride and 0.40 g (4.0 mmol) of triethylamine are added. The mixture is left to stand at room temperature for two days. Evaporation and flash chromatography on silica gel with CH2Cl2: MeOH (9: 1) gives 0.13 g (60%) of the desired compound. RMN-1 !! (600 MHz, CDC13): d 1.80 (m, 1H), 1.93 (m, 1H), 2.08 (m, 1H), 2.20 (s, 3H), 2.27 (m, 1H), 3.22 (rt ?, 1H) , '3.65-3.8 (m, 5H, a singlet thereof at 3.74 ppm), 3.80 (s, 3H), 4.06 (m, 1H), 4.32 (dd, 1H), 4.38 (dd, 1H), 4.51 (, 1H), 4.71 (m, 1H), 4.93 (broad, 2H), 6.78 (dd, 1H), 6.81 (d, 1H), 6.85 (d, 1H), 7.08 (d, 1H), 7.16 (d, 1H) ), 7.20 (s, 1H), 7.81 (broad t, 1H) 13 C-NMR (100 MHz, CDC13) carbonyl carbons and amidine: d 174.7, 172.8, 154.9 (ix) (2, 5-diMeO) Ph- (i. or S) CH (CH2OH) -CO-Pro-Pab (2-Me) Dissolves (2, 5-diMeO) Ph- (R or 5) CH (CH20H) -CO-Pro-Pab (2-Me) (OH) (65 mg, 0.13 mmol, from step (viii) above) in 5 ml of ethanol. HOAc (8 drops of a Pasteur pipette) and 40 mg of 10% Pd / C are added. The hydrogenated. reaction mixture at atmospheric pressure for two days. The reaction mixture is filtered through celite and evaporated. The residue in water, washed with ethyl acetate and lyophilized. Yield: 46 mg (65%). 2 H NMR (500 MHz; D20) rotamers: d 1.8-2.1 (m, 6H, a singlet thereof at 1.92 ppm), 2.2-2.4 (m, 4H, two singles thereof to 2.24 (minor rotamer) and 2.38 ( major rotamer), 3.35 (m, 1H, minor rotamer), 3.6-3.7 (m, 1H), 3.75-3.85 (m, 7H, three singles thereof to 3.77, 3.79 and 3.84 ppm (rotamers)), 4.0- 4.1 (m, 1H), 4.4-4.5 (m, 3H), 4.8 (m, 1H partially obscured by the HDO peak), 6.80 (d, 1H, major rotamer), 6.85 (d, 1H, minor rotamer), 6.9 -7.1 (m, 2H), 7.45 (d, 1H, major rotamer), 7.51 (d, 1H, minor rotamer), 7.55-7.65 (m, 2H) NMR-: 3C (75 MHz, D20) carbonyl carbons and amidine: d 175.4, 174.5 (minor), 174.2 (minor), 174.1, 167.1 Example 3 The title compounds of Examples 1 and 2 are tested in Test A above and both are found to exhibit an IC50TT value of less than 0.3 μM.

Abbreviations aq = aqueous Bn = benzyl DCC = dicyclohexylcarbodiimide DIPEA diisopropylethylamine DMAP = N, N-dimethylaminopyridine DMF = dimethylformamide DMSO = dimethyl sulfoxide EDC = 1- (3-dimethylaminopropyl) 3-ethylcarbodiimide hydrochloride Et = ethyl EtOAc ethyl acetate EtOH = ethanol h = hours HBTU = [N, N, N ', N' tetramethyl-O- (benzotriazol-1-yl) uronium hexafluorophosphate HOAC = acetic acid H-Pab = 4-amidinobenci sheet H-Pab (Z) = 4- (N-benzyloxycarbonylamino) benzylamine H-Pab (2-Ci; = 4-amidino-2-chloro-benzylamine H-Pab (2-Me! = 4-amidino-2-methyl-benzylamine HPLC = high performance liquid chromatography Me = methyl MeOH = methanol Ph = phenyl Pr = propyl i-PrOH i-propanol TBTU = [N, N, N ', N' tetramethyl-O- (benzotriazol-1-yl) uronium tetrafluoroborate] TEA = triethylamine THF = tetrahydrofuran Z = benzyloxycarbonyl Prefixes n, s, i and t have their usual meanings: normal, iso, sec and tertiary. The stereochemistry for the amino acids is in default (S) if not established otherwise. It is noted that in relation to this date, the best method known by the applicant to carry out the said invention is that which is clear from the present description of the invention.

Claims (15)

CLAIMS Having. described the invention as above, claimed as property contained in the following claims:

1. A compound of formula I, characterized in that R1 represents H, CIOJR11, SiR12R13R14 or alkyl of 1 to 6 carbon atoms (in which the latter group is optionally substituted or terminated by one more substituents selected from OR 15 (CH2) qRi6); Ri, RIJ and R1 * independently represent H, phenyl or alkyl of 1 to 6 carbon atoms; R16 represents alkyl of 1 to 4 carbon atoms, phenyl, OH, C (0) OR17 or C (0) N (H) R18; R18 represents H, alkyl of 1 to 4 carbon atoms or CH2C (0) OR19; R15 and R17 independently represent H, alkyl of 1 to 6 carbon atoms or alkylphenyl of 7 to 9 carbon atoms; R 11 and R independently represent H alkyl of 1 to 4 carbon atoms; and q represents 0, 1 or 2; R2 and R3 independently represent H, alkyl of 1 to 4 carbon atoms, cyclohexyl or phenyl; Rx represents a structural fragment of the formula bundle, Ilb or lie, lia llb lie where k, 1 and m independently represent 0, 1, 2, 3 or 4; R4 and R5 independently represent H, If (Me) 3, 1- or 2-naphthyl, a polycyclic hydrocarbyl group, CHR 1R42 or alkyl of 1 to 4 carbon atoms (in which the latter group is optionally substituted by one or more substituents halo), or cycloalkyl of 3 to 8 carbon atoms, phenyl, methylenedioxyphenyl, benzodioxanyl, benzofuranyl, dihydrobenzofuranyl, benzothiazolyl, benzoxazolyl, benzimidazolyl, coumaranonyl, coumarinyl or di-idro-coumarinyl (in which the latter twelve groups are optionally substituted by one or more of alkyl of 1 to 4 carbon atoms (in which the latter group is optionally substituted by one or more halo substituents), alkoxy of 1 to 4 carbon atoms, halo, hydroxy, cyano , nitro, S02NH2, C (0) OH or N (H) R43); R41 and R42 independently represent cyclohexyl or phenyl; R6 and R7 independently represent H, alkyl of 1 to 4 carbon atoms, cycloalkyl of 3 to 8 carbon atoms, phenyl (in which the latter group is optionally substituted by one or more of alkyl of 1 to 4 carbon atoms ( in which the latter group is optionally substituted by one or more halo substituents), C 1 -C 4 alkoxy, halo, hydroxy, cyano, nitro, S02NH2, C (0) OH or N (H) R44 or together with 'the carbon atom to which they are attached form a cycloalkyl ring of 3 to 8 carbon atoms; Rq and R independently represent H R 45 represents H, alkyl of 1 to 4 carbon atoms or alkoxy of 1 to 4 carbon atoms; Y represents CH2, (CH2) 2, CH = CH, (CH2) 3. CH2CH = CH or CH = CHCH2, in which the last three groups are optionally substituted by alkyl of 1 to 4 carbon atoms, methylene, oxo or hydroxy; n represents 0, 1, 2, 3 or 4; and B represents a structural fragment of the formula IVa, IVb, IVe or IVd, IVa IVb IVe IVd where xJ X 'x- and X4 independently represent CH, N or N-O; X5 and X6 independently represent a single bond or CH2; one of X7, X8 and X9 represents S, O or NH, and the other two independently represent -CH =, = CH-, -N =, = N-, -N (0) = o = N (0) -; R 31 represents, in all cases, one or more optional substituents selected from halo, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms or -O- (CH 2) pC (O) N (R32) ( R33); p represents 0, 1, 2, 3 or 4; and R32 and R33 independently represent H, alkyl of 1 to 6 carbon atoms or cycloalkyl of 3 to 7 carbon atoms; or a pharmaceutically acceptable salt thereof.

2. A compound of the formula I, according to claim 1, characterized in that, when B represents a structural fragment for the formula IVa (in which X1, X2, X3 and X4 all represent CH), a structural fragment of the formula or IVb or a structural fragment of the formula or IVe, R31 represents one or more substituents selected from halo, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms or -0- (CH2) pc (O) N (R32) (R33).

3. A compound of the formula I, according to claim 1 or claim 2, characterized in that B represents a structural fragment for the formula IVa.

4. A compound of the formula I, according to any of the preceding claims, characterized in that the fragment is in the S configuration

5. A compound of the formula la, characterized in that Ba represents a structural fragment of the formula IVe, IVf, IVg or IVh IVe IVf IVg IV wherein D1 and D2 independently represent, in each case, H, OH, ORa, 0C (0) Rb, OC (0) ORc, C (0) 0Rd, C (0) Re, and Ra, Rb, Rc, Rd and Re independently represent alkyl of 1 to 12 carbon atoms (in which the latter group is optionally interrupted by oxygen and / or substituted by halo), phenyl, naphthyl, alkylphenyl of 1 to 3 carbon atoms (in which three groups they are optionally substituted by alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms, nitro or halo) or - (C (Rf) (Rg)) 2OC (O) C (Rh), Rf, Rg and Rh independently represent H or alkyl of 1 to 4 carbon atoms, and R1, R2, R3, Rx, Y, n, X1, X2, X3, X4, X5, X6, 'X7, X8, X9 and R31 are how defined in the foregoing, or a pharmaceutically acceptable salt thereof, with the proviso that D1 and D2 both do not represent H.

6. A pharmaceutical formulation, characterized in that it includes a compound according to any of the preceding claims, or a pharmaceutically acceptable salt thereof, in admixture with a pharmaceutically acceptable adjuvant, diluent or carrier.

7. A compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, for use as a pharmaceutical.

8. A compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, for use in the treatment of a condition where the inhibition of thrombin is required or desired.

9. A compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, for use in the treatment of thrombosis.

10. A compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, for use as an anticoagulant.

11. The use of a compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, as an active ingredient in the manufacture of a medicament for the Treatment of a condition where inhibition of thrombin is required or desired.

12. The use according to claim 11, characterized in that the condition is thrombosis.

13. The use according to claim 11, characterized in that the condition is hypercoagulability in the blood and tissues.

14, The use of a compound according to any of claims 1 to 5, or a pharmaceutically acceptable salt thereof, as an active ingredient in the manufacture of an anticoagulant.

15. A process for the preparation of a compound of the formula I, characterized in that it comprises: (a) the coupling of a compound of the formula V, wherein R1, R2, R3 and Rx are as defined in claim 1 with a compound of the formula VI, wherein Y, n and B are as defined in claim 1; (b) coupling a compound of formula VII, wherein R1, R2, R3, Rx and Y are as defined in claim 1 with a compound of formula VIII, H2N- (CH2) "-B VIII wherein R1, R ', R3 and Rx are as defined in claim 1 with a compound of the formula VI, wherein Y, n and B are as defined in claim 1; (b) coupling a compound of formula VII, wherein R1, Rr, R3, Rx and Y are as defined in claim 1 with a compound of formula VIII, H2N- (CH2) n-B VIII wherein n and B are as defined in claim 1; (c) for compounds of the formula I in which B represents a structural fragment of the formula IVa, IVb or IVe, reaction of a compound of the formula IX, where B "represents a structural fragment of the formula IVa :, IVb: o ¡Ve '' ^ otf IVa1 fVb1 I Ve1 wherein Ry represents alkyl of 1 carbon atoms and R1, R2 R3, R? Y / n. X1 X 'X x < x- x ( R 31 are as defined in claim 1, with ammonium gas; (d) for compounds of the formula I in which B represents a structural fragment of the formula IVa, IVb or IVe, reduction of a compound of the formula X, wherein B represents a structural fragment of the formula IVa2, IVb2 or IVe2 IVa2 IVb2 IVe2 and R1, R2, R3, Rx, Y, n, X1, X2, X3, X4, X5, X6 and R.3"1 are as defined in claim 1, or (e) for compounds of the formula I, wherein X1, X2, X3, X4, X7, X8 and / or X9 represents NO, oxidation of a corresponding compound of the formula I in which X1, X2, X3, X4, X7, X8 and / or X9 (as appropriate) represents N. ^ 3 NEW PHARMACEUTICAL COMPOUNDS USEFUL AS SELECTIVE INHIBITORS OF SERINE PROTEASES SUMMARY OF THE INVENTION Compounds of the formula (I) are provided, wherein R1, R2, R3, Rx, Y, n and B have the meanings given in the description, which are useful as competitive inhibitors of proteases such as trypsin, such as thrombin, and in particular in the treatment of conditions where the inhibition of thrombin is required (for example, thrombosis) or as anticoagulants.