MXPA00000911A - Oligo-thiophenes useful as antimetastatic agents, a preparation thereof and pharmaceutical compositions containing them - Google Patents

Abstract

Oligo-thiophenes of formula (I) are disclosed wherein:A is a -CH2-CO-, -CH2CH2-CO- or -CH=CH-CO- group;B is a 2-thienyl or a phenyl group, which is substituted by an R group, or it is a natural or synthetic amino acid, linked to the A group via its N-terminus;or A-B is a -CH2CH2-NH-A.A.- group, wherein the A.A.-group is a natural or synthetic amino acid residue, linked to the CH2CH2-NH-group via its carboxy-terminus;R is hydrogen or a chlorine, bromine, iodine, fluorine, (C1-C4)alkyle, (C1-C4)alkylene- COOR', (C1-C4)alkylene-NH2, (C1-C4)alkylene-NR'2 or (C1-C4)alkylene-NHCOR'group;R'is hydrogen or a (C1-C4)alkyl group, isomers thereof, and salts thereof with pharmaceutically acceptable acids and bases. It is also claimed the use of the compounds of formula (I) as inhibitors of the uPA binding to the specific uPAR receptor, in particular their application as antitumor and antimetastatic agents.

Description

OLIGO-TIOFENOS USEFUL AS ANTIMETASTATIC AGENTS, A PREPARATION OF THEM AND THE COMPOSITIONS PHARMACEUTICALS THAT CONTAIN THEM FIELD OF THE INVENTION The present invention relates to oligo-thiophenes, optionally linked to natural or synthetic amino acids. It has been found that these compounds possess uPA-uPAR antagonist activity and can be used with antitumor and / or antimetastatic agents.

BACKGROUND OF THE INVENTION Serine proteases uPA (urokinase-type plasminogen activator) catalyzes the activation of plasminogen to plas ina, which is involved in a variety of physiological and pathological processes. uPA is a multi-domain protein that has a * B "catalytic chain (amino acids 144-411) and an iso-terminal fragment (* ATF", aa 1-143) comprised of a domain similar to growth factor (aa 4) -43) and a kringle domain (aa 47- REF ": 32621 135). uPA is a multifunctional protein involved in tissue proteolysis, cell migration, cell proliferation and growth factor activation. uPA is released from cells as a virtually inactive, pro-uPA proenzyme. Activation of the single-stranded pro-uPA by plasmin (which leads to the active form of two chains) is regulated by strict control mechanisms that are not yet fully understood. Most uPA activities are confined to the cell surface and the pericellular environment. This is carried out by the binding to a specific receptor of high affinity on the cell surface (uPAR). Both forms of uPA are linked to uPAR with similar affinity. The binding interaction is mediated by the domain similar to the growth factor [S.A. Rabbani et al., J. Bi ol. Ch em. 267, 14151-56, 1992]. The uPA receptor is a three-domain glycoprotein where each triplicate portion comprises a consensual sequence rich in cysteine of approximately 90 amino acids [M. Plow et al. J. Bi ol. Chem. , 268, 17539-46, 1993]. uPAR is anchored to the cell membrane by a glucosyl-phosphatidylinositol moiety (GP1 anchor). uPAR is linked to uPA with KD values between 10 ~ 10 and 10 ~ 9 M, depending on the experimental system. The major determinants for uPA binding are located in the N-terminal 1 domain, uPAR can be excised by uPA and plasmin, releasing a water-soluble domain 1 and by the action of phospholipase C, three uPAR domains (1 + 2 +3) can be released from the cell surface. This latter form of uPAR is also soluble in water because the GPl anchor is missing. The inhibition of uPA-dependent phenomena can be mainly focused in two ways, either by direct inhibition of proteolytic activity or by inhibition of uPA receptor binding. The latter strategy has the potential to achieve greater specificity, since the inhibition can be localized to the pericellular environment. A technique of bacteriophage visualization and protein engineering have recently been used to discover peptide and site-specific uPAR antagonists [Goodson et al. PNAS, 91, 7129, 1994; Stratton-Thomas et al., Prot. Eng. , 5, 463-470, 1995, respectively].

The present invention relates to oligo-thiophenes with potent antagonistic activity. Many derivatives of bistiofen and tertiophene appear to show interesting biological properties [Kagan J. et al., J. Org. Chem. 48, 4317-20, 1983 and references cited therein]. Most notably these are toxic to nematodes and this effect can be greatly increased by the presence of ultraviolet light. The most carefully studied of these compounds is a-tertienyl, a natural product derived from plants first recognized as phototoxin in the 70s. This compound showed photo-increased activity against nematodes, microorganisms, algae, human erythrocytes, insect larvae and eggs, as well as of the generation of pigmentation in the skin, acting as a herbicide and as an inhibitor of seed germination. Antiviral and cytotoxic, photoactive activities were also reported [(a) Cooper et al., Bi oorg. Ch em. , 13, 362-374, 1985 and references cited therein; (b) Rawis et al., Ch em. & Engin News, 21-23, 1986; (c) Evans et al., J. Am. Chem. Soc. , 112, 2694-2701, 1990; (d) Kyo et al., Pl an t Cel l Rep. , 9, 393-397, 1990; (e) Hudson et al., Pl an ta Med. , 59, 447-450, 1993; (f) Hudson et al., Che osph ere, 19, 1329-1343, 1989]. A structure-activity relationship study was announced for the fourteen unsubstituted isomeric tertiofennes [Jayasuriya et al., Heterocycl, 24, 2261-2264 and 2901-2904, 1986]. The use of the thiophene ring is very widespread in the different fields of therapeutic agents, whereas oligothiophenes are only rarely mentioned as pharmacological agents [Press et al., The Chemi stry of He terocycl is Compounds, vol. 44, part four, Chapter III, Edited by Salo Grono itz 396-502, 1989]. The uPA / uPAR system has been shown to be involved in a variety of invasive biological processes such as tumor metastasis, trophoblast implantation, inflammation and angiogenesis. Therefore, uPAR antagonists should be able to block tumor invasiveness, metastasis and angiogenesis. Formulations containing uPAR antagonists represent novel therapeutic treatments for a number of highly invasive and metastatic cancers, where it has been found that uPA and uPAR are consistently present in the invading tumor foci [Daño et al., Proteolysi s and Protein Turnover, eds. . Barret + Bond, Portlan Press, 1994, London] (for example, cancers of the breast, lung, colon, ovaries). In patients with breast cancer and non-small cell lung cancer, increased levels of uPAR have been detected in plasma. Therefore, the amount of soluble uPAR seems to reflect the degree of proteolysis in the tumor and this may be closely related to the patient's prognosis. The levels of uPA and uPAR in the tumor tissue are prognostic factors in many types of cancers. In addition to cancer, other diseases mediated by uPA cell surface activity are attacked by uPAR antagonists. The inhibitors of the generation of plasmin by the uPA bound to the receptor, therefore have tumoristic, anti-invasive, antimetastatic, antiangiogenic, antiarthritic, anti-inflammatory, anti-osteoporotic, antiretinoptic and contraceptive activities, based on the mechanism. These compounds are preferably applied via the oral route, but also by intravenous or intramuscular injections, nasal sprays or any other conventionally used application.

DESCRIPTION OF THE INVENTION The present invention relates to the oligo-thiophenes of the general formula (I): wherein: A is a group -CH2CO-, -CH2CH2-CO- or -CH = CH-CO-; B is a 2-thienyl group or a phenyl group, which is substituted by a group R, or this is a natural or synthetic amino acid, linked to group A via its N-terminus; or A-B- is a group -CH 2 CH 2 -NH-A.A-, wherein the group A.A. is a natural or synthetic amino acid residue linked to the CH2CH2-NH group via its carboxyl terminus; R is hydrogen or a chlorine atom, bromine, iodine, fluorine, (alkyl of 1 to 4 carbon atoms), (alkylene of 1 to 4 carbon atoms) -COOR '; (alkylene of 1 to 4 carbon atoms) -NH2, (alkylene of 1 to 4 carbon atoms) -NR '2 or (alkylene of 1 to 4 carbon atoms) -NHCOR '; - R 'is hydrogen or an alkyl group of 1 to 4 carbon atoms, the isomers thereof, and the salts thereof with pharmaceutically acceptable acids and bases.

A natural amino acid denotes one of the 20 a-amino acids which are the monomeric units for the polypeptides, for example glycine, alanine, valine, leucine, isoleucine, phenylalanine, tryptophan, proline, serine, threonine, tyrosine, aspartic acid, glutamic acid, asparagine, glutamine, lysine, arginine, histidine, cysteine or methionine. A synthetic amino acid consists of a compound comprising an amino group together with a carboxyl group bonded to an α-carbon atom. Pharmaceutically acceptable salts of the compounds of the present invention include the physiologically acceptable acid addition salts for example hydrochlorides, hydrobromides, sulfates, methansulphonates, p-toluenesulfonates, phosphates, acetates, citrates, succinates, lactates, tartrates, fumarates and maleates . The salts can also be formed with bases, for example the sodium, potassium, magnesium, and calcium salts. Preferred compounds are those in which B is an amino acid or A-B is a group of the formula -CH 2 CH 2 -NH-A.A. Other preferred compounds are those in which B is a 2-thienyl group. Particularly preferred compounds are those in which the amino acid is tryptophan. Another object of the present invention is to provide a process for the preparation of the compounds of the formula (I). A further objective of the present invention is the use of the compounds of the formula (I) for the treatment of diseases in which a uPAR inhibitor can be effective, in particular in tumors and tumor metastasis or tumor invasiveness, as well as the pharmaceutical compositions containing a pharmacologically effective amount of one or more compounds of the formula (I) in admixture with pharmaceutically suitable additives.

PREPARATION OF THE COMPOUNDS OF THE INVENTION The compounds of the formula (I) in which B is a natural or synthetic amino acid (hereinafter referred to as -A.A-) can be prepared starting from the intermediate of the formula (II): which is first transformed into the corresponding acyl chloride derivative and then this is reacted via the Friedel-Craft reaction with the intermediate of the formula (III): COOR "wherein A 'is a group -CH2-, -CH2CH2- or -CH = CH- and R" is an alkyl group of 1 to 4 carbon atoms, in the presence of a Lewis acid. The transformation of the intermediate (II) into its acyl chloride is preferably carried out with thionyl chloride, which is also used as the solvent, at temperatures between room temperature and the boiling temperature of the solvent. The Friedel-Craft reaction is carried out in an inert solvent and at a temperature in the range of -5 ° C to room temperature, preferably using SnCl 4 as the Lewis acid. The intermediary (IV) obtained as follows: is hydrolyzed in the corresponding acid of the formula (V): Suitable reaction conditions comprise the use of a base, preferably an alkali metal hydroxide, in water, an alcohol or mixtures thereof, at a temperature in the range of 0 ° C to room temperature. The carboxylic group of the intermediate (V) is then reacted, for example with the carbonyldiimidazole to give the imidazolyl derivative, and subsequently reacted preferably in an inert solvent at a temperature between 0 ° C and 50 ° C, with an amino acid of the formula -AA-, to give the intermediaries of the formula (V (): R "which are then converted to the corresponding compounds of the formula (I) via hydrolysis of the carboxylic ester -COOR", preferably under acidic conditions. If in the previous process the intermediary of the formula (III) is replaced by an intermediary of the formula (III '): the compounds of the formula (I) in which A-B is a group -CH 2 CH 2 -NH-A.A. - they can be prepared. In this case, the protective group -COCF3 must be eliminated after the Friedel-Craft reaction and the intermediate obtained thus must be condensed with an N-BOC-amino acid, previously activated through its carboxyl group. The compounds of the formula (I) are finally obtained after removal of the BOC protecting group. The compounds of the formula (I) in which B is a 2-thienyl group or a phenyl group, which is substituted with a group R having the above meanings, can be prepared by Friedel-Craft reaction of the intermediary of the Formula (II), activated in its carboxylic functional group as described above, with an intermediate of the formula (VII): wherein A has the meanings shown above and B 'is a 2-thienyl group or a phenyl group, which is substituted with a group R as shown above. The intermediates of the formula (II) can be obtained by reaction of a bis-thiophene, substituted on a ring with the R substituents, with carbon dioxide and a strong base such as butyllithium, at a temperature between -50 ° C. and -20 ° C. Bis-thiophene starting products are commercially available or can be prepared according to procedures well known to a person skilled in the art.

The intermediaries of the formula (VII) wherein A is -CH2-CO- or -CH = CH-CO- can be obtained starting from the commercially available intermediates of the formula (VIII): wherein A "is -CH2- or -CH = CH-, via the transformation of the carboxylic functional group into its corresponding acyl chloride and the subsequent Friedel-Craft reaction with a compound of the formula BH, wherein BH is a group thiophene or phenyl substituted with a group R as shown above.

The intermediates of the formula (VII) in which A is -CH2CH2-CO- can be obtained from the corresponding intermediates with A = -CH = CH-CO- via the hydrogenation of the C = C double bond in the presence of a suitable catalyst, such as (PPh3) 3RhCl. Analogously, the intermediates of the formula (III) in which A 'is -CH2CH2- can be obtained from the corresponding intermediates possessing C = C via the catalytic hydrogenation.

BIOLOGICAL ACTIVITY The compounds of the invention were tested (ELISA test) as inhibitors of human urokinase (uPA) which binds to its specific uPAR mAk receptor (BIO-R4), according to the procedure described in Biol. Chem. Hoppe-Seyler, 376 587-94 (1995) by Rettenberger et al. The assays are performed in 96-well microtiter plates. The following solutions are used: wash buffer: PBS buffer (without Mg2 + and Ca2 +) + 0.05% Tween 20; Incubation buffer (IP): 1% skimmed milk powder in PBS buffer (without Mg2 + and Ca2 +); BI0-R4 solution: 50 ng / well (0.5 μg / ml, 100 μl / well) in IP; uPAR solution: 3 ng / well (30 ng / ml; 100 μl / well) in PBS buffer (without Mg2 + and Ca2 +); blocking solution: 1% skimmed milk powder in wash buffer (dissolved at 37 ° C); uPA solution: 0.25 ng / well (5 ng / ml, 50 μl / well) in IP. Detection solutions (per microtiter plate): (1) 6 ml (100 mM Tris-Cl pH 7.2 + 0.15% Tween 80) + 1.5 ml (10 μg) of plasminogen in bidistilled water; (2) 6 ml (100 mM Tris-Cl pH 7.2 + 0.15% Tween 80) + 1.5 ml (7.5 mg) of Chromozyme PL in bidistilled water. The detection solution must be continuously stirred. Test substances: the test substances are dissolved in DMSO. These are used in the test system with a higher concentration of 100 μg / ml. The solutions are prepared using PBS. Three controls are carried out: a) positive control: using 2% DMSO in PBS; b) negative control: assay without receptor; c) control of inhibition: 1) inhibition (IC95 at 0.25 mg / ml) with dextran sulfate (molecular weight = 500,000); 2) inhibition (IC90 at 1 μg / ml) with inactivated uPA (175 μg / ml). Incubation is carried out as follows: Each well incubated with 100 μl of BI0-R4 (c = 0.5 μg / ml) for 1 hour at room temperature under agitation. After washing three times with the wash buffer, each well is incubated for 1 hour at 37 ° C with 200 μl / well of blocking solution. After triple washing, each well is incubated for 1 hour at room temperature under agitation with 100 μl / well of uPAR (c = 30 ng / ml), then the wells are again washed three times with the wash buffer. The solution of the test substance and the control solution, respectively, are added (50 μl / well) and incubated for 30 minutes at room temperature under agitation. An additional 50 μl of uPA solution (c = 2.5 ng / ml) is added.

After 1 hour at room temperature, a triple wash is carried out. For detection, the following procedure is used: Incubation with 50 μl of each of the detection solution (1) and (2) at room temperature. After 20 minutes a yellow color will be visible (the positive control reads an extinction of 1 after 45 to 60 minutes.) Detection is performed at 405 nm (reference is 490 nm) using a Dynatech MR 7000 ELISA reader. the percentage of inhibition the following formula is used (E means extinction): % inhibition = 1 00 - 100 X [E-test-Econtrol neg. / Econtrol pos - Econtrol neg. J The data for a representative compound of the invention are reported in table I.

Table I - assay BIO-R4 - inhibition of uPA binding to the specific uPAR receptor (BI0-R4) expressed as IC5o (mM) The invention relates to pharmaceutical agents that contain one or more compounds of the formula (I). In order to produce pharmaceutical agents, the compounds of the formula (I) are mixed in a known manner with suitable carrier, aromatic, flavoring and pharmaceutical dyes, and are for example formed into tablets or coated or suspended tablets. or they are dissolved in water or in an oil such as olive oil with the addition of appropriate auxiliary substances The substance of the general formula (I) can be administered orally or parenterally in a liquid or solid form. medium containing stabilizing agents, solubilizers and / or buffers, which are usually used for solutions for injection Such additives are for example tartrate or borate buffers, ethanol, dimethyl sulfoxide, complexing agents (such as ethylenediaminetetraacetic acid) , high molecular weight polymers (such as liquid polyethylene oxide) for regulation of the viscosity or polyethylene derivatives of sorbitol anhydrides. The solid carrier substances are, for example, starch, lactose, mannitol, methylcellulose, talc, highly dispersed silicic acid, higher molecular weight fatty acids (such as stearic acid), gelatin, agar-agar, calcium phosphate, stearate magnesium, animal and vegetable fats or high molecular weight solid polymers (such as polyethylene glycols). Formulations suitable for the oral route may, if desired, contain flavorings and sweeteners. The dose administered depends on the age, health and weight of the patient, the degree of the disease, the type of treatments that are possibly being carried out concurrently, the frequency of the treatment and the type of the desired effect. The daily dose of the active compound is usually 0.1 to 50 mg / kg of body weight. Normally 0.5 to 40 and preferably 1 to 20 mg / kg / day in one or several applications per day are effective in order to obtain the desired results. The invention is further illustrated by the following examples: Preparation Synthesis of bis-thiophene-2-carboxylic acid Dissolve 15 g of bis-thiophene in 300 ml of tetrahydrofuran (THF) and the solution is cooled to -40 ° C under nitrogen atmosphere. 10.8 ml of 10 M butyl lithium in hexane are carefully added dropwise, then the red solution is kept at -10 ° C for 1 hour and emptied into a 500 g dry ice flask (solid carbon dioxide). The yellow suspension is dissolved in 1 hour, then the resulting solution is poured into 10 ml of 2 N sodium hydroxide. After 30 minutes, the organic phase is separated and the solvent is concentrated to a small volume. To the residue is added 2N sodium hydroxide and water and washed with hexane, then the aqueous phase is acidified to pH = 2 with 37% hydrochloric acid, maintained for 1 hour at room temperature, and the solid is filtered and dried. Wash with 50 ml of water. After drying it in an oven at 70 ° C, 17.8 g of the product are obtained, m.p. 171-174 ° C.

Preparation Synthesis of 2- [3- (2-thienyl) acryloyl] thiophene A solution of 5 g of 3- (2-thienyl) acrylic acid in 30 ml of thionyl chloride is heated at 50 ° C for 1 hour 30 minutes, then the unreacted thionon chloride is removed under reduced pressure. To the residue is added 20 ml of heptane and dried twice, then dissolved in 50 ml of methylene chloride, cooled to 0 ° C and a solution of 2.43 g of thiophene in 5 ml of water is added dropwise. methylene chloride. Maintaining the temperature at 0 ° C, a solution of 6.74 ml of SnCl 4 in 12 ml of methylene chloride is then added. The reaction mixture is kept at room temperature for 2 hours, then it is drained into 200 ml of 2N hydrochloric acid. The aqueous phase is separated and extracted with 100 ml of methylene chloride. The organic phase is dried over sodium sulphate and evaporated to dryness and the residue is purified by chromatography on silica gel (eluent: hexane / ethyl acetate 5: 1), obtaining, after recrystallization from hexane, 4.79 g. of the product, pf 105-107 ° C.

Preparation Synthesis of 2- [3- (2-thienyl) propanoyl] thiophene A solution of 4.64 g of 2- [3- (2-thienyl) -acyloyl] thiophene in 50 ml of anhydrous methylene chloride and 0.97 g of tris (triphenylphosphine) rhodium chloride is hydrogenated at room temperature overnight. The reaction mixture is then concentrated to dryness and purified by chromatography on silica gel (eluent hexane / ethyl acetate 20: 1) to give 4.3 g of the product as an oil.

Preparation 4. Synthesis of 2- [2- ((2,2 '-bistien-5' -yl) carbonyl) thien-5-yl] acetic acid ethyl ester A solution of bis-thiophene-2-carboxylic acid (3 g, preparation 1) in 20 ml of thionyl chloride is heated at 50 ° C for 1 hour 30 minutes, then the unreacted thionyl chloride is evaporated and the residue is evaporated. Treat three times with 10 ml of heptane and evaporate. The residue is dissolved in methylene chloride and cooled to 0 ° C, then a solution of 2.19 g of 2- (thien-2-yl) acetic acid ethyl ester in 5 ml of chloride is added dropwise thereto. of methylene. Subsequently 3 ml of an SnCl solution is subsequently added. in 10 ml of methylene chloride. The reaction mixture is kept at room temperature for 2 hours, then it is drained in 200 ml of 2N hydrochloric acid. The aqueous phase is extracted with methylene chloride (3x150 ml) and the organic extracts are collected, dried over sodium sulfate, sodium and concentrate to dryness. The residue (8 g) is purified by chromatography on silica gel (eluent hexane / ethyl acetate 8: 1) to give 3.62 g of the product.

Preparation 5. Synthesis of 2- [2 - ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] acetic acid A suspension of 3.62 g of 2- [2- ((2,2'-bis-thien-5'-yl) -carbonyl) thien-5-yl] -acetic acid ethyl ester in 60 ml of ethanol and 5 ml of water, it is cooled to 0 ° C and 11.35 ml of 2 N sodium hydroxide are added. The mixture is kept at room temperature overnight, then it is concentrated to a small volume. After acidification, the product crystallizes, which is then separated by filtration and dried under vacuum at 40 ° C. 2.4 g of the product are recovered.

Preparation 4- [3- (2-thienyl) propanolyl] -1-fluorobenzene g of the 2- (2-thienylidene) -4-fluoroacetophenone are dissolved in 50 ml of anhydrous methylene chloride and 0.97 g of tris (triphenylphosphine) rhodium chloride are added. The reaction is hydrogenated for 8 hours (about 470 ml of hydrogen are reacted), then concentrated to dryness and purified by chromatography on silica gel (eluent hexane / ethyl acetate 20: 1), to give 4.85 g of the product .

Preparation 2- [2- (trifluoroacetamido) -1-ethyl] thiophene A solution of 10 g of 2- (2-thienyl) ethylamine in 100 ml of tetrahydrofuran is cooled to 0 ° C, then 9.05 ml of ethyl trifluoroacetate are added dropwise. After stirring for 4 hours at room temperature, the reaction mixture is concentrated to dryness to give 18 g of an oil which is treated with 18 ml of ethyl ether and saturated with 90 ml of hexane. After cooling for 2 hours, 12.15 g of the product are collected by filtration, m.p. 45-47 ° C.

Preparation 2- [2- ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] -N-trifluoroacetylethylamine A solution of bis-thiophene-2-carboxylic acid (3.4 g, preparation 1) in 25 ml of thionyl chloride was heated at 50 ° C for 2 hours. The reaction mixture was concentrated to dryness, then redissolved in heptane and concentrated again to dryness. This work is repeated, then the residue is dissolved in methylene chloride, cooled to 0 ° C and a solution of 3.3 g of 2- [2- (trifluoroacetamido) -1-ethyl] thiophene is added dropwise (preparation 7) in 20 ml of methylene chloride and subsequently with a solution of 3.8 ml of SnCl__ in 3.8 ml of methylene chloride. The reaction mixture is kept under stirring at room temperature for 2 hours, then it is drained in 200 ml of 2N hydrochloric acid and extracted with tetrahydrofuran (4 x 100 ml). The organic extracts are combined, dried over sodium sulfate and the solvent is evaporated under reduced pressure. 18 g of a black oil are obtained, which is purified by chromatography on silica gel (eluent hexane / tetrahydrofuran 2: 1). After recrystallization from "hexane (100 ml) and drying under vacuum at 50 ° C, 4.3 g of the product are obtained, mp 168-170 ° C.

Preparation 9. 2- [2- ((2,2 '-bistien-5' -yl) carbonyl) thien-5-yl] ethylamine To a mixture of 2- [2- ((2, 2'-bis-thien-5'-yl) carbonyl) thien-5-yl] -N-trifluoroacetylethylamine (10 g, preparation 8) in 800 ml of methanol, cooled at 0 ° C and kept under stirring, 60.17 ml of 2 N sodium hydroxide are added and the reaction mixture is heated at 50 ° C overnight. The mixture is then concentrated to a small volume, then 500 ml of water are added and extracted with 4 portions of 200 ml of chloroform. The organic extracts are combined, dried over sodium sulfate and concentrated to dryness. After crystallization from a mixture of ethyl acetate (16 ml) / hexane (16 ml), 5.1 g of the product, m.p. 102-104 ° C.

Example 1. Synthesis of ethyl ester N- [2- [2- ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] acetyl] tryptophan A suspension of 2.34 g of 2- [2- ((2, 2'-bistien-5'-yl) carbonyl) thien-5-yl] acetic acid (preparation 5) and 1.74 g of carbonyldiimidazole in 150 ml of anhydrous tetrahydrofuran it is maintained at 40 ° C for 1 hour. The mixture was cooled to 20 ° C and a solution of 2.45 g of tryptophan ethyl ester in 10 ml of anhydrous tetrahydrofuran was added dropwise thereto. After 2 hours the reaction mixture is concentrated to dryness, then dissolved in ethyl acetate, washed with water and extracted with 200 ml of chloroform. The organic phase is concentrated to dryness to give 7 g of a residue which is purified by chromatography on silica gel (eluent: hexane / ethyl acetate from 4: 1 to 2: 1). 2.79 g of the product are obtained.

Example 2. Synthesis of N- [2- [2- ((2,2 '-bistien-5' -yl) carbonyl) thien-5-yl] acetyl] tryptophan A solution of 2.2 g of N- [2- [2 - ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] acetyl] tryptophan ethyl ester (example 1) in 320 ml of Ethanol and 10 ml of water is cooled to 0 ° C, then 4.43 ml of 2 N sodium hydroxide is added dropwise. After 4 hours at room temperature, the mixture is concentrated to a small volume, water is added it is washed with ethyl acetate and acidified to pH = 2. The aqueous phase is extracted with ethyl acetate, the organic extracts are dried over sodium sulfate, concentrated to dryness and the residue is purified by chromatography on silica gel. (eluent chloroform / methanol / acetic acid 9: 1: 0.25) to give 1.16 g of the product, mp 166-168 ° C.

Example 3 Synthesis of 2- [3- [2- ((2,2 '-bistien-5' -yl) carbonyl) thien-5-yl] propanoyl] thiophene A solution of 1 g of bis-thiophene-2-carboxylic acid (preparation 1) in 5 ml of thionyl chloride is heated at 50 ° C for 1 hour 30 minutes, then the unreacted thionyl chloride is evaporated and the residue is evaporated. He adds heptane and evaporates again. The residue is dissolved in methylene chloride, cooled to 0 ° C and a solution of 2- [3- (2-thienyl) propanoyl] thiophene (1 g; preparation 3) in 3 ml of anhydrous methylene chloride. A solution of 1.05 ml of SnCl4 in 3 ml of anhydrous methylene chloride is then added dropwise, maintaining the temperature at 0 ° C. The reaction mixture is kept at room temperature for 1 hour, then it is emptied into 100 ml of acid 2 N hydrochloric acid, extracted with ethyl acetate (3 x 100 ml), dried over sodium sulfate and concentrated to dryness. 3 g of the residue are purified by chromatography on silica gel (eluent: hexane / ethyl acetate 5: 1) to give 1.02 g of the product, m.p. 124-126 ° C.

Example 4. Synthesis of 2- [3- [2- ((2,2 '-bistien-5' -yl) carbonyl) thien-5-y1] acryloyl] thiophene A solution of 0.24 g of bis-thiophene-2-carboxylic acid (preparation 1) in 3 ml of thionyl chloride was heated at 50 ° C for 1 hour 30 minutes, then the unreacted thionyl chloride was evaporated and the residue was evaporated. He adds heptane and evaporates again. The residue is dissolved in methylene chloride, cooled to 0 ° C and a solution of 2- [3- (2-thienyl) acryloyl] thiophene (0.3 g, preparation 2) in 3 ml of chloride is added dropwise. of anhydrous methylene. A solution of 0.31 ml of SnCl in 3 ml of anhydrous methylene chloride is then added dropwise, keeping the temperature at 0 ° C. The reaction mixture is kept at room temperature for 4 hours, then an additional 0.3 ml of SnCl is added and the mixture is kept at room temperature overnight. The reaction mixture is poured into 200 ml of 2N hydrochloric acid, extracted with 3 portions of 50 ml of ethyl acetate, dried over sodium sulfate and concentrated to dryness. 1.3 g of the residue is purified by chromatography on silica gel (eluent hexane / ethyl acetate 5: 1, then pure ethyl acetate) to give 0.37 g of the product.

Example 5. Synthesis of 2- [3- [2- ((2,2 '-bistien-5' -yl) carbonyl) thien-5-yl] propanoyl] -4-fluorobenzene 1. 06 g of the bis-thiophene-2-carboxylic acid (preparation 1) in 10 ml of thionyl chloride is heated at 50 ° C for 1 hour 30 minutes, then the mixture is concentrated to dryness. The residue is treated twice with 20 ml of heptane, finally with 30 ml of methylene chloride, cooled to 0 ° C and a solution of 4- [3- (2-thienyl) propanoyl] -1 is added dropwise. -fluorobenzene (1.08 g, preparation 6) in 5 ml of methylene chloride. Maintaining the temperature at approximately 0 ° C, a solution of SnCl4 (1.07 ml) in 5 ml of methylene chloride is added dropwise, then the mixture is stirred at room temperature by. 2 hours. The reaction mixture is poured into 150 ml of 2 N hydrochloric acid, then extracted with ethyl acetate and the organic phase is concentrated to dryness. The residue is purified by chromatography on silica gel (eluent methylene chloride), to give after recrystallization from ethyl acetate / hexane, 1 g of the product, m.p. 138-140 ° C.

Example 6 According to the procedures described in the previous preparations and the previous examples, the following oligo-thiophene derivatives are obtained: - N- [2- [2- ((2,2 '-bistien-5'-yl) carbonyl) thien-5-yl] -acetyl] glycine; N- [2- [2 - ((2, 2'-bistien-5'-yl) carbonyl) thien-5-yl] -acetyl] alanine; N- [2- [2 - ((2, 2'-bistien-5'-yl) carbonyl) thien-5-yl] -acetyl] phenylalanine; N- [2- [2 - ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] acetyl] tyrosine; N- [2- [2 - ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] acetyl] leucine; - N- [2- [2- ((2,2 '-bistien-5' -yl) carbonyl) thien-5-yl] acetyl] lysine; N- [2- [2 - ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] acetyl] serine; N- [2- [2 - ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] acetyl] isoleucine; 2- [3- [2 - ((2, 2'-bistien-5'-yl) carbonyl) thien-5-yl] propanoyl] -4-fluorobenzene; 2- [3- [2 - ((2,2 '-bistien-5' -yl) carbonyl) thien-5-yl] propanoyl] -4-chlorobenzene; - 2- [3- [2- ((2, 2'-bistien-5'-yl ') carbonyl) thien-5-yl] propanoyl] -2-bromobenzene; 2- [3- [2 - ((2,2 '-bistien-5' -yl) carbonyl) thien-5-yl] propanoyl] -4- (carboxymethyl) benzene; 2- [3- [2 - ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] propanoyl] -4- (aminomethyl) benzene; 2- [3- [2 - ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] propanoyl] -3- (dimethylaminomethyl) benzene; 2- [3- [2 - ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] propanoyl] -2- (tert-butyl) benzene; 2- [3- [2 - ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] acryloyl] -4-fluorobenzene; 2- [3- [2- ((2, 2'-bistien-5 '-yl) carbonyl) thien-5-yl] acryloyl] -4-chlorobenzene; 2- [3- [2 - ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] acryloyl] -2-bromobenzene; 2- [3- [2 - ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] acryloyl] -4- (carboxymethyl) benzene; 2- [3- [2- ((2,2 '-bistien-5' -yl) carbonyl) thien-5-yl] acryloyl] -4- (aminomethyl) benzene; 2- [3- [2- ((2,2 '-bistien-5' -yl) carbonyl) thien-5-yl] acryloyl] -3- (dimethylaminomethyl) enne; 2- [3- [2- ((2, 2'-bisti en-5 '-yl) carbonyl) thien-5-yl] acryloyl] -2- (tert-butyl) benzene.

Use 4 - [2- [2- ((2,2'-bistien-5i il) carbonyl) thien-5-yl] ethylaminocarbonyl] -2- (tert-butoxycarbonylaminomethyl) butanoic acid A mixture of glutamic acid N-BOC (0.88 g), and 0.63 g of 1,1 '-carbonyldiimidazole in 40 ml of tetrahydrofuran is kept under stirring at room temperature for 2 hours, then a solution of 2- is added dropwise. [2- ((2,2 '-bistien-5' -yl) carbonyl) thien-5-yl] ethylamine (1.11 g, preparation 9) in 40 ml of tetrahydrofuran and the reaction mixture is heated at 50 ° C all the night. The solvent is evaporated under reduced pressure and the residue is dissolved in ethyl acetate and washed with a saturated aqueous solution of potassium acid sulfate. After separation by filtration of the separating solid (initial product), the organic phase is concentrated to dryness. The residue (2.5 g) was purified by chromatography on silica gel (eluent 1) chloroform / methanol 15: 2: 2) chloroform / methanol (9: 1), to give 0.42 g of the product.

For example Acid 4- [2- [2 - ((2,2 '-bistien-5' yl) carbonyl) thien-5-yl] ethylaminocarbonyl] -2-aminobutane A suspension of 4- [2- [2- ((2,2 '-bistien-5'-yl) carbonyl) thien-5-yl] ethylaminocarbonyl] -2- (tert-butoxycarbonylamino) butanoic acid (0.28 g; 7) 0.39 ml of trifluoroacetic acid are added in 28 ml of methylene chloride. The resulting dark solution is kept under stirring at room temperature overnight, then it is concentrated to dryness. The residue is redissolved in 15 ml of absolute ethanol, then concentrated again to dryness and this treatment is repeated once more. 0.3 g of the residue was suspended in 20 ml of hexane, stirred for 1 hour at room temperature, filtered and dried under vacuum at 30 ° C, 0.2 g of the product was obtained. X H NMR in dβ-DMSO + D 20: 1.9 ppm (m, 2H); 2.18 ppm (, 2H); 3.05 ppm (m, 2H); 3.36 ppm (broad m, 2H); 3.7 ppm (t, ÍH); 7.1 ppm (m, 2H); 7.5 ppm (m, 3H); 7.9 ppm (m, 2H); 8.61 ppm (broad t, ÍH).

Example 9 According to the methods described in Examples 7 and 8, starting from the appropriate amino acids, the following oligo-thiophenes are obtained: [2- [2- ((2, 2'-bistien-5'-yl) carbonyl ) thien-5-yl] -ethylamino] leucinamide; - [2- [2- ((2,2 '-bistien-5' -yl) carbonyl) thien-5-yl] -ethylamino] phenylalaninamide; [2- [2 - ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] -ethylamino] serinamide; [2- [2- ((2, 2'-bistien-5'-yl) carbonyl) thien-5-yl] -ethylamino] lysinamide; [2- [2 - ((2,2 '-bistien-5' -yl) carbonyl) thien-5-yl] ethylamino] isoleucinamide; [2- [2 - ((2,2 '-bistien-5' -yl) carbonyl) thien-5-yl] ethylamino] histidinamide; [2- [2 - ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] ethylamino] tyrosinamide.

It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention.

Claims (9)

CLAIMS Having described the invention as above, the content of the following claims is claimed as property:

1. The oligo-thiophenes of the general formula I): characterized in that: A is a group -CH2CO-, -CH2CH2-CO- or -CH = CH-CO-; B is a 2-thienyl group or a phenyl group, which is substituted by a group R, or this is a natural or synthetic amino acid, linked to group A via its N-terminus; or A-B- is a group -CH2CH2-NH-A.A-, wherein the group A.A.- is a natural or synthetic amino acid residue, linked to the group CH2CH2-NH via its carboxyl terminus; R is hydrogen or a chlorine atom, bromine, iodine, fluorine, (alkyl of 1 to 4 carbon atoms), (alkylene of 1 to 4 carbon atoms) -COOR '; (C 1 -C 4 -alkylene) -NH 2, (C 1 -C 4 -alkylene) -NR '2 or (C 1 -C 4 -alkylene) -NHCOR'; R 'is hydrogen or an alkyl group of 1 to 4 carbon atoms, the isomers thereof, and the salts thereof with pharmaceutically acceptable acids and bases.

2. The oligo-thiophenes according to claim 1, characterized in that B is an amino acid.

3. The oligo-thiophenes according to claim 2, characterized in that A-B is a group of the formula -CH 2 CH 2 -NH-A.A-.

4. The oligo-thiophenes according to claims 1 to 3, characterized in that the amino acid is tryptophan.

5. The oligo-thiophenes according to claim 1, characterized in that B is a 2-thienyl group.

6. The oligo-thiophenes according to claim 1, characterized in that they are selected from: N- [2- [2 - ((2,2 '-bistien-5' -yl) carbonyl) thien-5-yl] -acetyl] tryptophan; - 2- [3- [2- ((2, 2'-bistien-5'-yl) carbonyl) thien-5-yl] -propanoyl] thiophene; 2- [3- [2 - ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] -acyloyl] thiophene; 2- [3- [2 - ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] -propanoyl] -4-fluorobenzene; N- [2- [2 - ((2,2 '-bistien-5' -yl) carbonyl) thien-5-yl] -acetyl] glycine; N- [2- [2 - ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] -acetyl] alanine; - N- [2- [2- ((2, 2'-bistien-5'-yl) carbonyl) thien-5-yl] -acetyl] phenylalanine; N- [2- [2- ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] -acetyl] tyrosine; N- [2- [2 - ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] -acetyl] leucine; N- [2- [2 - ((2, 2'-bistien-5'-yl) carbonyl) thien-5-yl] -acetyl] lysine; N- [2- [2- ((2,2 '-bistien-5'-yl) carbonyl) thien-5-yl] -acetyl] serine; - N- [2- [2- ((2, 2'-bistien-5'-yl) carbonyl) thien-5-yl] -acetyl] isoleucine; 2- [3- [2 - ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] -propanoyl] -4-fluorobenzene; 2- [3- [2 - ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] -propanoyl] -4-chlorobenzene; 2- [3- [2 - ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] -propanoyl] -2-bromobenzene; 2- [3- [2 - ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] -propanoyl] -4- (carboxymethyl) benzene; - 2- [3- [2- ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] -propanoyl] -4- (aminomethyl) benzene; 2- [3- [2 - ((2,2 '-bistien-5' -yl) carbonyl) thien-5-yl] -propanoyl] -3- (dimethylaminomethyl) benzene; 2- [3- [2 - ((2, 2'-bistien-5'-yl) carbonyl) thien-5-yl] -propanoyl] -2- (tert-butyl) benzene; 2- [3- [2 - ((2,2 '-bistien-5' -yl) carbonyl) thien-5-yl] -acyloyl] -4-fluorobenzene; 2- [3- [2 - ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] -acryloyl] -4-chlorobenzene; 2- [3- [2 - ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] acryloyl] -2-bromobenzene; 2- [3- [2 - ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] acryloyl] -4- (carboxymethyl) benzene; 2- [3- [2 - ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] acryloyl] -4- (aminomethyl) enne; 2- [3- [2 - ((2, 2'-bistien-5'-yl) carbonyl) thien-5-yl] acryloyl] -3- (dimethylaminomethyl) benzene; 2- [3- [2 - ((2,2'-bistien-5'-yl) carbonyl) thien-5-yl] acryloyl] -2- (tert-butyl) benzene.

7. Pharmaceutical compositions containing a pharmacologically effective amount of one or more compounds according to claims 1 to 6, in admixture with pharmaceutically acceptable excipients.

8. The use of the compounds according to claims 1 to 6, for the preparation of a medicament having anti-tumor and anti-metastatic activity. 9 The compounds according to claims 1 to 3, for use as inhibitors of uPA binding to the specific uPAR receptor.