HK1048314B

HK1048314B - 2-arylimino-2,3-dihydrothiazoles, and their use thereof as somatostatin receptor ligands

Info

Publication number: HK1048314B
Application number: HK03100526.0A
Authority: HK
Inventors: Moinet Christophe; Sackur Carole; Thurieau Christophe
Original assignee: Ipsen Pharma S.A.S.
Priority date: 1999-07-22
Filing date: 2000-07-21
Publication date: 2009-08-21

Description

2-arylimino-2, 3-dihydrothiazoles and their use as somatostatin receptor ligands

The subject of the present application is novel 2-arylimino-2, 3-dihydrothiazole derivatives and processes for their preparation. These products have good affinity with certain subtypes of somatostatin receptors and thus have useful pharmacological properties. The invention also relates to these products as medicaments, to pharmaceutical compositions containing them and to their use for the preparation of medicaments intended for the treatment of pathological conditions or diseases in which one (or more) somatostatin receptor(s) is (are) involved.

Growth hormone release inhibitor (SST) is a cyclic tetradecapeptide, a growth hormone-inhibiting substance that was first isolated from the hypothalamus (Brazeau P et al, science 1973, 179, 77-79). It also acts as a neurotransmitter in the brain (Reisine T. et al neuroscience 1995, 67, 777-. Molecular cloning has shown that the biological activity of somatostatin is directly dependent on the family of five receptors attached to the membrane.

Studies on the heterogeneity of biological functions of somatostatin have attempted to determine the structure-activity relationships of peptide analogs at somatostatin receptors, and thus 5 subtypes of receptors have been discovered (Yamada et al, Proc. Natl. Acad. Sci. USA 89, 251-255, 1992; Raynor, K. et al, mol. Pharma. 44, 385-392, 1993). The utility of these receptors is currently being actively studied. The affinity to different subtypes of somatostatin receptors has been linked to the treatment of the following disorders/diseases. Activation of subtypes 2 and 5 has been associated with inhibition of Growth Hormone (GH), more particularly with inhibition of GH-secreting adenomas (acromegaly) and hormone-secreting TSH. Activation of subtype 2, but not subtype 5, has been associated with the treatment of prolactin secreting adenomas. Other indications associated with the activation of somatostatin receptor subtypes are recurrence of stenosis, inhibition of insulin and/or glucagon secretion, in particular diabetes, hyperlipidemia, insulin insensitivity, syndrome X, vascular diseases, proliferative retinopathy, dawn phenomenon and nephropathy; inhibition of gastric acid secretion, in particular peptic ulcers, enterocutaneous and pancreaticocutaneous fistulas, irritable colon syndrome, dumping syndrome, watery diarrhea syndrome, AIDS-associated diarrhea, chemotherapy-induced diarrhea, acute or chronic pancreatitis and secretory gastrointestinal tumors; treatment of cancers such as hepatoma; inhibition of angiogenesis, treatment of inflammation such as arthritis; chronic allograft rejection; performing angioplasty; prevention of bleeding from transplanted blood vessels and gastrointestinal bleeding. Somatostatin agonists can also be used to reduce the body weight of a patient.

Among the pathological disorders associated with somatostatin (Moreau J.P. et al, Life sciences 1987, 40, 419; Harris A.G. et al, European journal of medicine 1993, 2, 97-105), mention may be made, for example, of: acromegaly, pituitary adenoma, Cushing's disease, gonadotropinoma and prolactinoma, glucocorticoid catabolic side effects, insulin-dependent diabetes mellitus, diabetic retinopathy, diabetic nephropathy, hyperthyroidism, gigantism, endocrine gastroenteropancreatic tumors (including carcinoid syndrome, pancreatic adenoma, insulinoma, insulinomatosis, hyperinsulinemia, glucagonomas, gastrinoma and Zollinger-Ellison syndrome), growth hormone releasing factor tumor of esophageal varices and acute hemorrhage, gastroesophageal reflux, gastroduodenal reflux, pancreatitis, refractory diarrhea of enterocutaneous and pancreatic fistulas, acquired immunodeficiency syndrome, chronic secretory diarrhea, diarrhea associated with irritable bowel syndrome, disorders associated with gastrin-releasing peptide, secondary lesions of intestinal transplantation, portal hypertension and variceal hemorrhage in cirrhosis patients, Gastrointestinal bleeding, gastroduodenal ulcer bleeding, crohn's disease, systemic sclerosis, dumping syndrome, small bowel syndrome, hypotension, scleroderma and medullary thyroid carcinoma, diseases associated with cellular hyperproliferation (such as cancer, more precisely breast cancer, prostate cancer, thyroid cancer, and pancreatic and colorectal cancers), fibrosis (more precisely renal fibrosis, hepatic fibrosis, pulmonary fibrosis, skin fibrosis, central nervous system fibrosis and nasal fibrosis and fibrosis induced by chemotherapy) and other therapeutic areas, such as headache (including headache associated with pituitary tumours), pain, panic attacks, chemotherapy, wound healing, renal insufficiency caused by development, obesity and developmental retardation associated with obesity, uterine developmental retardation, skeletal dysplasia, noonan syndrome, sleep apnea syndrome, cerebral infarction, cerebral hemorrhage, cerebral infarction, graves 'disease, ovarian polycystic disease, pancreatic pseudocysts and ascites, leukemia, meningioma, cancer cachexia, inhibition of helicobacter pylori, psoriasis and Alzheimer's disease. Osteoporosis may also be mentioned.

Applicants have found that the compounds of formula (I) below have affinity and selectivity for somatostatin receptors. Since somatostatin and its peptide analogs often have poor oral bioavailability and poor selectivity (Robinson, C. (future drugs) 1994, 19, 992; Reubi, J.C. et al TIPS 1995, 16, 110), said compounds are non-peptide agonists or antagonists of somatostatin and can be advantageously used in the treatment of the above-mentioned and pathological conditions or diseases in which one (or more) somatostatin receptor(s) is/are involved. Preferably, the compounds can be used for the treatment of acromegaly, pituitary adenoma, or endocrine gastroenteropancreatic tumors including carcinoid syndrome.

The compounds of the present invention are of the general formula (I)

Or a racemate, enantiomeric forms or a combination of all of these forms of (a), wherein:

r1 represents amino (C)₂-C₇) Alkyl, aminoalkyl arylalkyl, aminoalkyl cycloalkylalkyl, (C)₁-C₁₅) Alkyl, (C)₃-C₇) Cycloalkyl group, (C)₁-C₆) Alkyl radical (C)₃-C₆) Cycloalkyl group, (C)₃-C₆) Cycloalkylalkyl, cyclohexenylalkyl, alkenyl, alkynyl, a carbocyclic aryl radical containing at least two rings, at least one of which is not an aromatic ring, a carbocyclic or heterocyclic aralkyl radical which may optionally be substituted on the aryl, bis-arylalkyl, alkoxyalkyl, furanylalkyl, tetrahydrofuranyl, dialkylaminoalkyl, N-acetamidoalkyl, cyanoalkyl, alkylthioalkyl, arylhydroxyalkyl, aralkyloxyalkyl, morpholinoalkyl, pyrrolidinylalkyl, piperidinoalkyl, N-alkylpyrrolidinylalkyl, N-alkylpiperazinylalkyl or oxopyrrolidinylalkyl radical,

or R1 represents one of the following radicals:

or R1 represents a-C (R11) (R12) -CO-R10 radical;

r2 represents an optionally substituted carbocyclic or heterocyclic aryl radical,

or R2 represents one of the following radicals:

r3 represents alkyl, adamantyl, an optionally substituted carbocyclic or heterocyclic aryl radical, an optionally substituted carbocyclic or heterocyclic aralkyl radical in an aryl group,

or R3 represents one of the following radicals:

or R3 represents a-CO-R5 radical;

r4 represents H, alkyl, carbocyclic or heterocyclic aralkyl optionally located on the aryl radical;

orThe radicals being of the formulaA radical in which i represents an integer from 1 to 3;

r5 represents a N (R6) (R7) radical;

r6 represents (C)₁-C₁₆) Alkyl, cycloalkylalkyl, hydroxyalkyl, aryloxyalkyl radical, optionally substituted carbocyclic or heterocyclic aralkyl radical on the aryl radical, aralkyloxyalkyl, arylhydroxyalkyl, alkoxyalkyl, alkylthioalkyl, alkenyl, alkynyl, cyclohexenyl, cyclohexenylalkyl, alkylthiohydroxyalkyl, cyanoalkyl, N-acetamidoalkyl radical, optionally substituted bis-arylalkyl radical on the aryl radical, optionally substituted di-arylalkyl radical on the aryl radical, morpholinoalkyl, pyrrolidinylalkyl, piperidinoalkyl, N-alkylpyrrolidinylalkyl, oxopyrrolidinylalkyl, tetrahydrofuranyl alkyl, N-benzylpyrrolidinylalkyl, N-alkylpiperazinylalkyl, N-benzylpiperazinylalkyl, N-benzylpiperidinylalkyl or N-alkoxycarbonyl piperidinyl radical,

or R6 represents (C) optionally substituted by a radical selected from the group consisting of₃-C₈) Cycloalkyl radical: a hydroxyl radical and an alkyl radical,

or R6 represents one of the following radicals:

r7 represents H or an alkyl, hydroxyalkyl, mono-or di-aminoalkyl or aralkyl radical;

or-N (R6) (R7) represents a radical of the formula:

wherein:

r8 represents H, alkyl, hydroxyalkyl, optionally substituted carbocyclic or heterocyclic aryl, optionally substituted aralkyl, alkenyl, alkoxyalkyl, cycloalkyl, cycloalkylalkyl, bis-arylalkyl, piperidinyl, pyrrolidinyl, hydroxy, arylalkenyl,

or R8 represents-X- (CH)₂)_b-R9；

R9 represents H or an alkyl, alkoxy, aryloxy, optionally substituted carbocyclic or heterocyclic aryl, morpholinyl, pyrrolidinyl, alkylamino or N, N' - (alkyl) (aryl) amino radical;

x represents CO, CO-NH or SO₂；

Y represents CH or N;

a represents 1 or 2;

b represents an integer from 0 to 6;

or the N (R6) (R7) radical represents a radical of the formula:

wherein:

z represents CH, O or S;

c represents an integer from 0 to 4;

or the N (R6) (R7) radical represents one of the following radicals:

r10 represents amino (C)₂-C₇) Alkylamino, ((aminoalkyl) aryl) alkylamino, ((aminoalkyl) cycloalkyl) alkylamino, piperazinyl, homopiperazinyl radical,

or R10 represents the following radical:

r11 represents H;

r12 represents H or alkyl, (C)₃-C₇) Cycloalkyl, optionally substituted carbocyclic or heterocyclic aralkyl, propargyl, allyl, hydroxyalkyl, alkylthioalkyl, arylalkyl alkoxyalkyl, arylalkylthioalkoxyalkyl radicals;

alternatively, the compounds of the present invention are salts of compounds of formula (I).

If the compounds of the general formula (I) contain R1, R2, R3, R4, R6, R8 or R9 radicals which comprise a substituted aryl radical or an aralkyl radical substituted on the aryl radical, the aryl or aralkyl radical is preferably such that:

-with respect to R1, if aryl is substituted, it may be substituted 1 to 5 times (except for the bond to the rest of the molecule) with radicals independently selected from the group consisting of: halogen atoms and alkyl, alkoxy, alkylthio, haloalkyl, haloalkoxy, aryl, aralkoxy or SO₂NH₂A radical of atoms. If appropriate, two substituents may be joined together to form a ring, for example together representing a methylenedioxy or propylene radical.

-with respect to R2, if aryl is substituted, it may be substituted 1 to 5 times (except for the bond to the rest of the molecule) with radicals independently selected from the group consisting of: halogen atoms and alkyl, alkoxy, alkylthio, haloalkyl, alkenyl, haloalkoxy, nitro, cyano, azido, SO₂N, mono-or di-alkylamino, aminoalkyl, aralkoxy or aryl radical. If appropriate, two substituents may be joined together to form a ring, for example together representing a methylenedioxy, ethylenedioxy or propylene radical.

With respect to R3, if the aryl group or groups (derived from aryl or aralkyl radicals) are substituted, they are substituted as appropriate from 1 to 5 times (except for the bond linking to the rest of the molecule). The carbocyclic aryl or aralkyl radical may be substituted 1 to 5 times on the aryl ring with radicals independently selected from the group consisting of: halogen atoms and alkyl, hydroxy, alkoxy, haloalkyl, haloalkoxy, nitro, cyano, azido, mono-or di-alkylamino, pyrrolidinyl, morpholinyl, aralkyloxy or aryl radicals. If appropriate, two substituents may be joined together to form a ring, for example together representing an alkylenedioxy radical having from 1 to 3 carbon atoms. The heterocyclic aryl or aralkyl radical of R3 may be substituted 1 to 2 times on the ring with radicals independently selected from the group consisting of: halogen atom and alkyl radical.

With respect to R4, if the aryl group is substituted, it may be substituted 1 to 5 times (except for the bond linking to the rest of the molecule). The aryl radical may be substituted with radicals independently selected from the group consisting of: halogen atoms and alkyl or alkoxy radicals.

With respect to R6, if the aryl group or groups are substituted, they may be substituted 1 to 5 times (except for the bond linking to the rest of the molecule). The optional substituents on the aryl group are independently selected from the group consisting of halogen atoms and alkyl, alkoxy, alkylthio, haloalkyl, haloalkoxy, aryl, aryloxy or SO₂NH₂Groups of radicals.

With respect to R8, if the aryl group or groups are substituted, they may be substituted 1 to 5 times (except for the bond linking to the rest of the molecule). The optional substituents on the aryl group are independently selected from the group comprising halogen atoms and alkyl, haloalkyl, alkoxy, hydroxy, cyano, nitro or alkylthio radicals.

With respect to R9, if a carbocyclic or heterocyclic aryl radical is substituted, it may be substituted 1 to 5 times (except for the bond to the rest of the molecule). Optional substituents on the aryl group are independently selected from the group comprising halogen atoms and alkyl, haloalkyl, alkoxy, haloalkoxy, alkylthio, carbocyclic aryl, hydroxy, cyano or nitro radicals.

With respect to R12, if a carbocyclic or heterocyclic aryl radical is substituted, it may be substituted 1 to 5 times (except for the bond to the rest of the molecule). Optional substituents on the aryl group are independently selected from the group comprising halogen atoms and alkyl, alkoxy, carbocyclic aryl, aralkoxy, hydroxy, cyano or nitro groups.

Unless otherwise specified, alkyl represents a straight or branched alkyl radical containing from 1 to 6 carbon atoms. Unless otherwise specified, cycloalkyl refers to a monocyclic carbon system containing 3 to 7 carbon atoms. Unless otherwise specified, alkenyl represents a straight or branched alkyl radical containing from 1 to 6 carbon atoms and at least one degree of unsaturation (double bond). Unless otherwise specified, alkynyl refers to a straight or branched alkyl radical containing from 1 to 6 carbon atoms and at least one double unsaturation (triple bond). Carbocyclic or heterocyclic aryl means a carbocyclic or heterocyclic ring system containing at least one aromatic ring, and if at least one of the rings containing it contains a heteroatom (O, N or S), this is referred to as a heterocyclic ring system. Haloalkyl represents an alkyl radical in which at least one hydrogen atom (optionally all) is replaced by a halogen atom.

Alkylthio, alkoxy, haloalkyl, haloalkoxy, aminoalkyl, alkenyl, alkynyl and aralkyl radicals denote, respectively, alkylthio, alkoxy, haloalkyl, haloalkoxy, aminoalkyl, alkenyl, alkynyl and aralkyl radicals in which the alkyl radical has the abovementioned meaning.

Straight-chain or branched alkyl having 1 to 6 carbon atoms denotes in particular the methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl and tert-butyl, pentyl, neopentyl, isopentyl, hexyl, isohexyl radical. Cycloalkyl denotes in particular cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl radicals. Carbocyclic or heterocyclic aryl denotes in particular phenyl, naphthyl, pyridyl, furyl, thienyl, 2, 3-indanyl, indolyl, imidazolyl, benzofuryl, benzothienyl, phthalimidyl radicals. Carbocyclic or heterocyclic aralkyl denotes in particular benzyl, phenethyl, phenylpropyl, phenylbutyl, indolylalkyl, phthalimidoalkyl, naphthylalkyl, furanylalkyl, thienylalkyl, benzothienylalkyl, pyridylalkyl and imidazolylalkyl radicals.

If an arrow is issued from the chemical structure, it represents a point of attachment. For example:

represents a benzyl radical.

Preferably, the compounds of formula (I) are such that:

r1 represents-C (R11) (R12) -CO-R10 or one of the following radicals:

r2 represents one of the following radicals:

r3 represents CO-R5 or one of the following radicals:

r4 represents H, alkyl, optionally substituted carbocyclic or heterocyclic aralkyl on the aryl radical,

r5 represents one of the following radicals:

r10 represents one of the following radicals:

r11 represents H;

r12 represents one of the following radicals:

it goes without saying that, with respect to R4, if the aryl group is substituted, it may be substituted 1 to 5 times (except for the bond linking to the rest of the molecule) by radicals independently selected from the following group: halogen atoms and alkyl or alkoxy radicals.

The compounds of the invention are preferably such that: r4 represents H.

More preferably, the compounds according to the invention have the general formula (II)

Wherein:

r1 represents one of the following radicals

R2 represents one of the following radicals

R3 represents one of the following radicals

R4 represents H;

or R1 represents one of the following radicals

R2 represents one of the following radicals

R3 represents a COR5,

r4 represents a hydrogen atom or a hydrogen atom,

r5 represents one of the following radicals

Or finally, R1 represents a radical-C (R11) (R12) -CO-R10, in which

R10 represents a radical

R11 represents a hydrogen atom or a hydrogen atom,

r12 represents a radical

R2 represents a radical

R3 represents a radical

R4 represents H.

Furthermore, the present invention relates to a process for the preparation of the aforementioned compounds of the general formula (I) (which is also applicable to the corresponding compounds of the general formula (II)) on a solid support.

According to the invention, compounds of the general formula (I) a

Wherein:

r1 represents-CH₂-A1-NH₂Radical, in which A1 represents- (CH)₂)_n-、-(CH₂)_n-O-(CH₂)_p-, aralkylene or cycloalkyleneAn alkyl radical, n and p represent integers from 1 to 6;

r2 and R4 represent the same radicals as in formula (I);

r3 represents the same radical as in formula (I) except for the-CO-R5 radical;

for example, it can be prepared according to a process characterized in that it comprises the following successive stages:

1) p-nitrophenylcarbonate Wang resin was treated with a large excess of R1-NH in an aprotic solvent, such as methylene chloride or dimethylformamide₂Symmetric diamine treatment;

2) treating the resin isolated after stage 1) with an aromatic isothiocyanate of the formula R2-N ═ C ═ S in an aprotic solvent, such as dichloromethane or dimethylformamide, wherein the R2 radical has the same meaning as in formula (I) a;

3) treating the resin obtained in stage 2) with a compound of formula (III) in an aprotic solvent, such as dioxane or dimethylformamide,

wherein the radicals R3 and R4 have the same meaning as in formula (I) a;

4) cracking the resin under acidic conditions;

5) treating the product obtained in stage 4) under alkaline conditions.

The preparation of p-nitrophenylcarbonate Wang resin is further described in the section "preparation of the Compounds of the invention".

Preferably, with respect to the above process, in order to achieve a large excess in stage 1), 10 to 20 equivalents of diamine R1-NH will be used₂. Stage 1) is preferably carried out at ambient temperature. Step (A)Stage 3) is carried out at a temperature above ambient temperature, for example at a temperature between 60 and 90 ℃, using 2 to 5 equivalents of the compound of formula (III). In stage 4), acidic conditions, which may be generated, for example, using a 50% dichloromethane/trifluoroacetic acid mixture, are preferably maintained for 1 to 2 hours. In stage 5), alkaline conditions can be generated, for example, using a saturated sodium bicarbonate solution or eluting on an alkaline alumina cartridge.

According to a variant of the invention, the compounds of the general formula (I) b

Wherein:

r1 represents the same radical as in formula (I) except-CH₂-A1-NH₂Other than type atomic group, wherein A1 represents- (CH)₂)_n-、-(CH₂)_n-O-(CH₂)_p-, aralkylene or cycloalkylalkylene radical, n and p represent integers from 1 to 6, with the exception of the-C (R11) (R12) -CO-R10 radical;

r2 represents an aminoalkylphenyl radical;

r3 represents the same radical as in formula (I) except for the-CO-R5 radical;

r4 represents the same atomic group as in the general formula (I);

1) p-nitrophenylcarbonate Wang resin is used with an excess of R2-NH in an aprotic solvent, such as dichloromethane or dimethylformamide₂Wherein the radical R2 has the same meaning as in formula (I)1 b;

2) treating the resin isolated after stage 1) with an isothiocyanate of the formula R1-N ═ C ═ S in an aprotic solvent, such as dichloromethane or dimethylformamide, wherein the R1 radical has the same meaning as in formula (I) b;

wherein the radicals R3 and R4 have the same meaning as in formula (I) b;

4) cracking the resin under acidic conditions;

5) treating the product obtained in stage 4) under alkaline conditions.

Preferably, with respect to the above process, in order to achieve an excess in stage 1), 5 to 10 equivalents of aminoalkylaniline will be used. Stage 1) is preferably carried out at ambient temperature. Stage 3) is carried out at a temperature above ambient temperature, for example at a temperature between 60 and 90 ℃, using 2 to 5 equivalents of the compound of formula (III). In stage 4), acidic conditions, which may be generated, for example, using a 50% dichloromethane/trifluoroacetic acid mixture, are preferably maintained for 1 to 2 hours. In stage 5), alkaline conditions can be generated, for example, using a saturated sodium bicarbonate solution or eluting on an alkaline alumina cartridge.

According to another variant of the invention, the compounds of the general formula (I) c

Wherein:

r1 represents-CH₂-A1-NH₂Radical, in which A1 represents- (CH)₂)_n-、-(CH₂)_n-O-(CH₂)_p-, aralkylene orA cycloalkyl alkylene radical, n and p represent integers from 1 to 6;

r2 represents the same atomic group as in the general formula (I);

r3 represents a-CO-R5 radical;

r4 and R5 represent the same radicals as in formula (I);

1) p-nitrophenylcarbonate Wang resin is washed with a large excess of a compound of formula R1-NH in an aprotic solvent, such as methylene chloride or dimethylformamide₂Wherein the radical R1 has the same meaning as in formula (I) c;

2) treating the resin isolated after stage 1) with an aromatic isothiocyanate of the formula R2-N ═ C ═ S in an aprotic solvent, such as dichloromethane or dimethylformamide, wherein the R2 radical has the same meaning as in formula (I) C;

3) treating the resin obtained in stage 2) with an acid of general formula (IV) in an aprotic solvent, such as dioxane or dimethylformamide,

wherein the radical R4 has the same meaning as in formula (I) c;

4) peptide coupling;

5) cracking the resin under acidic conditions;

6) treating the product obtained in stage 5) under alkaline conditions.

Preferably, with respect to the above process, in order to achieve a large excess in stage 1), 10 to 20 equivalents of symmetric diamine will be used. Stage 1) is preferably carried out at ambient temperature. Stage 3) is carried out at a temperature above ambient temperature, for example at a temperature between 60 and 90 ℃, using 2 to 5 equivalents of the acid of formula (IV). The peptide coupling of stage 4) is carried out, for example, in DMF with coupling agents such as Dicyclohexylcarbodiimide (DCC), Diisopropylcarbodiimide (DIC), DIC/N-hydroxybenzotriazole (HOBt) mixtures, benzotriazolyloxytris (dimethylamino) phosphonium hexafluorophosphate (PyBOP), 2- (1H-benzotriazol-1-yl) -1, 1, 3, 3-tetramethyluronium Hexafluorophosphate (HBTU) or 2- (1H-benzotriazol-1-yl) -1, 1, 3, 3-tetramethyluronium tetrafluoroborate (TBTU) and aminating compounds. Preferably, the coupling agent is used in a ratio of 4 to 5 equivalents, as is the aminating compound, and the reaction will be carried out at ambient temperature for 1 to 24 hours. In stage 5), acidic conditions, which may be generated, for example, using a 50% dichloromethane/trifluoroacetic acid mixture, are preferably maintained for 1 to 2 hours. In stage 6), alkaline conditions can be generated, for example, using a saturated sodium bicarbonate solution or eluting on an alkaline alumina cartridge.

According to another variant of the invention, the compounds of the formula (I) d

Wherein:

r1 represents the same atomic group as in the general formula (I) defined in claim 1, except-CH₂-A1-NH₂Other than type atomic group, wherein A1 represents- (CH)₂)_n-、-(CH₂)_n-O-(CH₂)_p-, aralkylene or cycloalkylalkylene radical, n and p represent integers from 1 to 6, with the exception of the-C (R11) (R12) -CO-R10 radical;

r2 represents an aminoalkylphenyl radical;

r3 represents a-CO-R5 radical;

r4 and R5 represent the same radicals as in formula (I);

can be prepared according to a process characterized in that it comprises the following successive stages:

1) p-nitrophenylcarbonate Wang resin is used with an excess of R2-NH in an aprotic solvent, such as dichloromethane or dimethylformamide₂Wherein the R2 radical has the same meaning as in formula (I) d;

2) treating the resin isolated after stage 1) with an isothiocyanate of the formula R1-N ═ C ═ S in an aprotic solvent, such as dichloromethane or dimethylformamide, wherein the R1 radical has the same meaning as in formula (I) d;

wherein the radical R4 has the same meaning as in formula (I) d;

4) peptide coupling;

5) cracking the resin under acidic conditions;

6) treating the product obtained in stage 5) under alkaline conditions.

Preferably, with respect to the above process, in order to achieve an excess in stage 1), 5 to 10 equivalents of aminoalkylaniline will be used. Stage 1) is preferably carried out at ambient temperature. Stage 3) is carried out at a temperature above ambient temperature, for example at a temperature between 60 and 90 ℃, using 2 to 5 equivalents of the acid of formula (IV). The peptide coupling of stage 4) is carried out, for example, in DMF with coupling agents such as Dicyclohexylcarbodiimide (DCC), Diisopropylcarbodiimide (DIC), DIC/N-hydroxybenzotriazole (HOBt) mixtures, benzotriazolyloxytris (dimethylamino) phosphonium hexafluorophosphate (PyBOP), 2- (1H-benzotriazol-1-yl) -1, 1, 3, 3-tetramethyluronium Hexafluorophosphate (HBTU) or 2- (1H-benzotriazol-1-yl) -1, 1, 3, 3-tetramethyluronium tetrafluoroborate (TBTU) and aminating compounds. Preferably, the coupling agent is used in a ratio of 4 to 5 equivalents, as is the aminating compound, and the reaction will be carried out at ambient temperature for 1 to 24 hours. In stage 5), acidic conditions, which may be generated, for example, using a 50% dichloromethane/trifluoroacetic acid mixture, are preferably maintained for 1 to 2 hours. In stage 6), alkaline conditions can be generated, for example, using a saturated sodium bicarbonate solution or eluting on an alkaline alumina cartridge.

According to another variant, the compounds of the general formula (I) e

Wherein:

r1 represents the same radical as in formula (I) except-CH₂-A1-NH₂Other than radicals, in which A1 represents- (CH)₂)_n-、-(CH₂)_n-O-(CH₂)_p-, aralkylene or cycloalkylalkylene radical, n and p represent integers from 1 to 6, with the exception of the-C (R11) (R12) -CO-R10 radical;

r2 represents the same atomic group as in the general formula (I);

r3 represents a-CO-R5 radical;

r4 represents H;

r5 represents-CH₂-A1-NH₂Radical in which A1 represents a linear or branched alkylene radical having 1 to 6 carbon atoms, - (CH)₂)_n-O-(CH₂)_p-, aralkylene or cycloalkylalkylene, N and p represent integers from 1 to 6, or R5 represents a N (R6) (R7) radical corresponding to the general formula:

wherein:

r8 represents H;

y represents N;

a represents 1 or 2;

1) p-nitrophenylcarbonate Wang resin is treated with a large excess of a symmetrical diamine of the formula R5-H in an aprotic solvent, such as dichloromethane or dimethylformamide;

2) peptide coupling with an acid of the general formula (IV) on the resin obtained in stage 1),

wherein the radical R4 has the same meaning as in formula (I) e;

3) general formula R1-NH₂Reacting a primary amine with an isothiocyanate of the formula R2-NCS in a solvent such as dimethylformamide or dioxane, R1 and R2 having the same meaning as in formula (I) e;

4) adding the thiourea obtained in the step 3) to the resin obtained in the step 2), and heating the mixture;

5) cracking the resin under acidic conditions;

6) treating the product obtained in stage 5) under alkaline conditions.

Preferably, with respect to the above process, in order to achieve a large excess in stage 1), 10 to 20 equivalents of diamine R5-NH will be used₂. Stage 1) is preferably carried out at ambient temperature. The peptide coupling of stage 2) is carried out with a coupling agent in DMF,coupling agents such as DIC/N-hydroxybenzotriazole (HOBt) mixtures. Preferably, the reaction of stage 3) is carried out in a solvent such as dimethylformamide or dioxane. During the addition of stage 4), 2 to 5 equivalents of thiourea will be used per equivalent of resin; preferably, the heating will be carried out at a temperature above ambient temperature, for example at a temperature of from 40 to 100 ℃ (precisely a temperature of about 80 ℃) for 2 to 24 hours. In stage 5), acidic conditions, which may be generated, for example, using a 50% dichloromethane/trifluoroacetic acid mixture, are preferably maintained for 1 to 2 hours. In stage 6), alkaline conditions can be generated, for example, using a saturated sodium bicarbonate solution or eluting on an alkaline alumina cartridge.

According to another variant of the invention, the compounds of formula (I) f

Wherein:

r1 represents a-C (R11R12) -CO-R10 radical;

r2, R3 and R4 represent the same radicals as in the general formula (I);

or R10 represents the following radical:

r11 represents H;

r12 represents H or alkyl, (C)₃-C₇) Cycloalkyl, optionally substituted carbocyclic or heterocyclic aralkyl, propargyl, allylHydroxyalkyl, alkylthioalkyl, arylalkyl alkoxyalkyl, arylalkylthioalkoxyalkyl radicals;

1) p-nitrophenylcarbonate Wang resin is treated with a large excess of a symmetrical diamine of the formula R10-H, wherein R10 has the same meaning as in formula (I) f, in an aprotic solvent, such as dichloromethane or dimethylformamide;

2) stage 1) peptide coupling of the resulting resin with amino acids of the general formula HOOC-C (R11) (R12) -NH-Fmoc, wherein R11 and R12 have the same meaning as in formula (I) f;

3) cleaving the Fmoc group of the resin obtained in stage 2);

4) stage 3) reaction of the resulting resin with an isothiocyanate of the formula R2-NCS, wherein R2 has the same meaning as in formula (I) f;

5) cracking the resin under acidic conditions;

6) treating the product obtained in stage 5) under alkaline conditions.

Preferably, with respect to the above process, in order to achieve a large excess in stage 1), 10 to 20 equivalents of diamine R10-H will be used. Stage 1) is preferably carried out at ambient temperature. The peptide coupling of stage 2) is carried out, for example, in DMF with coupling reagents such as Dicyclohexylcarbodiimide (DCC), Diisopropylcarbodiimide (DIC), DIC/N-hydroxybenzotriazole (HOBt) mixtures, benzotriazolyloxytris (dimethylamino) phosphonium hexafluorophosphate (PyBOP), 2- (1H-benzotriazol-1-yl) -1, 1, 3, 3-tetramethyluronium Hexafluorophosphate (HBTU) or 2- (1H-benzotriazol-1-yl) -1, 1, 3, 3-tetramethyluronium tetrafluoroborate (TBTU). Preferably, the reaction of stage 2) is carried out at ambient temperature for 1 to 24 hours. The deprotection of stage 3) can be carried out, for example, with the aid of a DMF mixture containing 20% piperidine. Stage 4) will preferably be carried out in a solvent such as dimethylformamide or dichloromethane, preferably with 5 to 10 equivalents of isothiocyanate added per equivalent of resin obtained in stage 3). In stage 5), acidic conditions, which may be generated, for example, using a 50% dichloromethane/trifluoroacetic acid mixture, are preferably maintained for 1 to 2 hours. In stage 6), alkaline conditions can be generated, for example, using a saturated sodium bicarbonate solution or eluting on an alkaline alumina cartridge.

The subject of the invention is also the compounds of the general formulae (I) and (II) indicated above or their pharmaceutically acceptable salts as medicaments. The invention also relates to pharmaceutical compositions containing said compounds or pharmaceutically acceptable salts thereof and to their use for the preparation of medicaments intended for the treatment of pathological conditions or diseases in which one (or more) somatostatin receptor(s) is (are) involved.

In particular, the compounds of the aforementioned general formulae (I) and (II) or a pharmaceutically acceptable salt thereof can be used for the preparation of a medicament intended for the treatment of a pathological condition or disease selected from the group consisting of: acromegaly, pituitary adenoma, Cushing's disease, gonadotropinoma and prolactin, catabolic side effects of glucocorticoids, insulin-dependent diabetes mellitus, diabetic retinopathy, diabetic nephropathy, syndrome X, dawn phenomenon, vascular disease, angioplasty, hyperthyroidism, gigantism, endocrine gastroenteropancreatic tumors (including carcinoid syndrome, pancreatic adenoma, insulinoma, insulinomatosis, hyperinsulinemia, glucagonoma, gastrinoma and Zollinger-Ellison syndrome), growth hormone releasing factor tumors of esophageal varices, as well as acute bleeding, ulceration, gastroesophageal reflux, gastroduodenal reflux, pancreatitis, intestinal skin and pancreatic fistula with diarrhea, refractory diarrhea of acquired immunodeficiency syndrome, chronic secretory diarrhea, diarrhea associated with irritable bowel syndrome, diarrhea induced by chemotherapy, Disorders associated with gastrin releasing peptide, secondary pathologies to intestinal transplantation, portal hypertension and variceal bleeding in patients with cirrhosis of the liver, gastrointestinal bleeding, gastroduodenal ulcer bleeding, graft vascular bleeding, crohn's disease, systemic sclerosis, dumping syndrome, small bowel syndrome, hypotension, scleroderma and medullary thyroid carcinoma, diseases associated with excessive cell proliferation (such as cancer, more precisely breast cancer, prostate cancer, thyroid cancer, and pancreatic and colorectal cancers), fibrosis (more precisely renal fibrosis, liver fibrosis, lung fibrosis, dermal fibrosis, fibrosis of the central nervous system and fibrosis of the nose and fibrosis induced by chemotherapy) and other therapeutic areas, such as headache, including headache associated with pituitary tumours, pain, inflammation (such as arthritis), panic attacks, chemotherapy, wound healing, pain relief, pain associated with inflammation (such as pain), and other therapeutic areas, Renal insufficiency caused by developmental delay, hyperlipidemia, obesity and obesity-related developmental delay, uterine developmental delay, skeletal dysplasia, noonan's syndrome, sleep apnea syndrome, graves ' disease, ovarian polycystic tumor disease, pancreatic pseudocysts and ascites, leukemia, meningioma, cancer cachexia, inhibition of helicobacter pylori, psoriasis, chronic allograft rejection and alzheimer's disease and osteoporosis.

Preferably, the compounds of the aforementioned general formulae (I) and (II) or a pharmaceutically acceptable salt thereof can be used for the preparation of a medicament intended for the treatment of a pathological condition or disease selected from the group consisting of: acromegaly, pituitary adenoma, or endocrine gastroenteropancreatic tumors (including carcinoid syndrome), and gastrointestinal bleeding.

Pharmaceutically acceptable salts mean in particular addition salts of inorganic acids, such as hydrochloride, sulfate, phosphate, hydrogen phosphate, hydrobromide and nitrate, or of organic acids, such as acetate, maleate, fumarate, tartrate, succinate, citrate, lactate, methanesulfonate, p-toluenesulfonate, pamoate, oxalate and stearate. Salts formed from bases such as sodium hydroxide or potassium hydroxide, where applicable, are also within the scope of the invention. As examples of other pharmaceutically acceptable salts, reference may be made to "pharmaceutically acceptable salts" journal of pharmaceutical science 66: 1(1977).

The pharmaceutical composition may be in solid dosage form, such as powder, granule, tablet, capsule, liposome or suppository. Suitable solid carriers may be, for example, calcium phosphate, magnesium stearate, talc, sugars, lactose, dextrin, starch, gelatin, cellulose, methyl cellulose, sodium carboxymethyl cellulose, polyvinylpyrrolidone and waxes. Suspensions contain, in particular, suspensions of slow-release microparticles loaded with the active ingredient (in particular microparticles of polylactide-co-glycolide or PLGA; see, for example, patent US 3,773,919, EP 52510 or EP 58481, or patent application pct wo 98/47489), it being possible to achieve a predetermined daily dose administered over a period of days to weeks.

Pharmaceutical compositions containing the compounds of the invention may also be in liquid form, for example as solutions, emulsions, suspensions or syrups. Suitable liquid carriers are, for example, water and organic solvents, such as glycerol or glycols, and mixtures thereof in water in various proportions.

The administration of the medicament of the present invention may be carried out by topical, oral, parenteral or intramuscular injection, etc.

The medicaments according to the invention are administered in doses of from 0.1mg to 10g, depending on the type of active compound used.

These compounds can be prepared as follows.

Preparation of the Compounds of the invention

I) Preparation of alpha-bromoketones

First method

This method is inspired by the scheme described in the following publications: macholan, l.; skursky, l. (chem. listy) 1955, 49, 1385-1388; bestman, h.j.; seng, F. (chemical report 1963, 96, 465-); jones, r.g.; kornfeld, e.c.; McLaughlin, K.C. Proc. USA society of chemistry 1950, 72, 4526-; nimgirawath, S.; ritchai, e.; taylor, W.C. J.Olympic Chemicals 1973, 26, 183-193.

The carboxylic acid is first converted to an acid with oxalyl chloride or thionyl chloride in the presence of a base (triethylamine or N-methylmorpholine) or activated to the anhydride form with an alkyl chloroformate (e.g. isobutyl chloroformate, see Krantz, A.; Copp, L.J.; biochemistry, 1991, 30, 4678-.

The activated carboxyl group is then converted to diazoketone with diazomethane in an ether solution or commercial solution of trimethylsilyldiazomethane (Aoyama, T.; Shiori, T.,. Chemicals bulletin, 1981, 29, 3249-3255) in an aprotic solvent, such as diethyl ether, Tetrahydrofuran (THF) or acetonitrile.

Bromination is then carried out with a brominating agent such as a solution of hydrobromic acid in acetic acid, aqueous hydrobromic acid in diethyl ether or dichloromethane.

Preparation example 1

2- (4-bromo-3-oxobutyl) -1H-isoindole-1, 3(2H) -dione

Oxalyl chloride (5.8 ml; 66.7mmol) was added to a solution of Pht-. beta. -Ala-OH (9.96 g; 44.5mmol) in dichloromethane (120ml) and 3 drops of Dimethylformamide (DMF). The mixture was stirred at ambient temperature for 3 hours. After removal of the solvent, the white solid was dissolved in a 1: 1 mixture of anhydrous tetrahydrofuran and acetonitrile (200ml), and 49ml of a 2M (trimethylsilyl) diazomethane in hexane (97.9mmol) was added dropwise at 0 ℃. After stirring overnight at 0 ℃, the solvent was removed. The pale yellow solid was then dissolved in dichloromethane (60ml) and 12ml aqueous hydrobromic acid (48%) was added dropwise at 0 ℃. The mixture was stirred until the temperature reached 15 ℃ and 50ml of saturated sodium bicarbonate solution were added. The organic phase was washed with brine and then dried over sodium sulfate. Crystallization from diethyl ether gave a white solid (11.39 g; yield 86%).

NMR ¹H(DMSO D6，100MHz，δ)：7.83(s，4H)；4.36(s，2H，CH₂Br)；3.8(t，2H，J＝7.1Hz，NCH₂)；2.98(t，2H，J＝6.9Hz，CH₂CO).

Preparation examples 2 to 11

The following compounds were prepared in a manner analogous to the procedure described in preparation 1:

^＊compounds which have been described in the literature

Second method

The starting product is an aryl ketone or a heteroaryl ketone.

The starting aryl or heteroaryl ketone is converted to the corresponding α -bromoketone using different brominating agents:

-CuBr₂(King, L.C.; Ostrum, G.K.; J. org. chem., 1964, 29, 3459-;

n-bromosuccinimide in CCl₄Or aqueous acetonitrile (Morton, H.E.; Leanna, M.R. & tetrahedron Kupflug)》1993，34，4481-4484)；

-bromine in glacial acetic acid or sulfuric acid;

phenyltrimethylammonium tribromide (Sanchez, J.P.; Parcell, R.P.; J.Chem, 1988, 25, 469-474), in an aprotic solvent, such as THF, or tetrabutylammonium tribromide (Kajigaeshi, S.; Kakinami, T.; Okamoto, T.; Fujisaki, S.; Japanese society of chemistry, 1987, 60, 1159-1160), in a dichloromethane/methanol mixture at ambient temperature;

brominating agents on polymeric supports, such as the perbromide poly (vinylpyridinium hydrobromide perbromide) on Amberlyst A-26 resin (Frechet, J.M.J.; Farrall, M.J.. J. Macro. Sci. 1977, 507-514), in aprotic solvents, such as methanol, at about 20-35 ℃ for about 2-10 hours.

Preparation example 12

1- (1-benzofuran-2-yl) -2-bromo-1-ethanone

A polymer of perbromide of pyridine hydrobromide (8.75 g; 17.5 mmol; 1.4 eq.) was added to a solution of (benzofuran-2-yl) methanone (2 g; 12.5mmol) in methanol (40 ml). The resulting mixture was stirred at ambient temperature for 7 hours and the reaction was terminated by filtration. Methanol was removed under reduced pressure and the expected product was crystallized by additional addition of diethyl ether (3.6 g; yield 60%).

NMR ¹H(DMSO D6，100MHz，δ)：8.09(s，1H)；7.98(d，1H，J＝6.6Hz)；7.75(d，1H，J＝8.4Hz)；7.58(t，1H，J＝8.4Hz)；7.4(t，1H，J＝7Hz)；4.83(s，2H，CH₂Br).

Preparation examples 8 to 12

The following compounds were prepared in a manner analogous to the procedure described in preparation 12:

^＊compounds which have been described in the literature

II) solid-phase Synthesis of 2-Arylimino-2, 3-dihydrothiazoles

Preparation of p-nitrophenyl carbonate Wang resin

The resin was prepared from Wang resin by a general procedure described in detail, the latter being obtained from Bachem or Novabiochem, with a load of more than 0.89mmol/g (see Bunin, B.A. "Combined index" Academic Press, 1998, p.62-63; Dressman, B.A.; span, L.A.; Kaldor, S.W. "tetrahedron Kuck." 1996, 37, 937. Ampere. 940; Hauske, J.R.; Dorff, P. "tetrahedron Kuck." 1995, 36, 1589. sup. 1592; Cao, J.; Cuny, G.D.; Hauske, J.R. "molecular diversity" 1998, 3, 173. sup. 179): n-methylmorpholine or pyridine as a base and 4-nitrophenyl chloroformate were added successively to Wang resin, the latter having been swollen beforehand at ambient temperature in Dichloromethane (DCM) or Tetrahydrofuran (THF). The mixture was stirred overnight. The resin was then washed successively with THF, diethyl ether and DCM and then dried overnight at 50 ℃ under reduced pressure.

Method A

Preparation of mono-protected symmetric diamines

General operation: para-nitrophenyl carbonate Wang resin is treated with a large excess of symmetrical diamine (10-20 equivalents) in an aprotic solvent such as DCM or DMF in order to produce a mono-protected diamine resin after stirring overnight as described in the literature (Dixit, D.M.; Leznoff, C.C.; J.C.S. Commission Chemicals 1977, 798-; Dixit, D.M.; Leznoff, C.C. J.C. J.S. J.Chem.252; KalJuste K.; Unden, A. tetrahedron Kupffer 1995, 36, 9211-; 9214; Munson, M.C.; Cook, A.W.; Josey, J.A. Rao, C. tetrahedron Kupffer 1998, 39, 7223-; 7226).

Preparation of thiourea resins

General operation: aromatic and heteroaromatic isothiocyanates (5-10 equivalents) are added to a mono-protected symmetric diamine in a solvent such as DCM or DMF and stirred overnight at ambient temperature (Smith, j.; Liras, j.l.; Schneider, s.e.; Anslyn, e.v.; journal of organic chemistry 1996, 61, 8811-8818). The thiourea resin was isolated by successive washings with DMF and DCM and then dried under reduced pressure at 50 ℃ overnight.

Preparation example 19

(Phenylthiocarbonyl) ethylcarbamate Wang resin

Phenyl isothiocyanate (1 ml; 8.5 mmol; 5eq.) was added to ethylenediamine N-carbamate Wang resin (2 g; 1.72 mmol; 0.86mmol/g) swollen in DCM (50 ml). After stirring at ambient temperature overnight, the resin was washed successively with DMF (5X 20ml) and DCM (5X 20 ml). The success of the coupling was monitored by means of the Kaiser ninhydrin test (Kaiser, E.; Colescott, R.L.; Bossinger, C.D.; Cook, P.I.; Ann. Biochemical Ann. 1970, 34, 595-598). A pale yellow resin (1.79g) was obtained with a loading of 0.648mmol/g, calculated from elemental analysis of sulfur.

Synthesis of 2-arylimino-2, 3-dihydrothiazoles

General operation: regioselective cyclization was carried out between thiourea resin and alpha-bromoketone (2-5 equivalents) in an aprotic solvent such as dioxane or DMF at 80 ℃ for 2-3 hours (Korohoda, M.J.; Bojarska, A.B.; J. Poland. chem. 1984, 58, 447-453; Ragab, F.A.; Hussein, M.M.; Hanna, M.M.; Hassan, G.S.; Kenawy, S.A.; J. Egypt. Pharman. J. Pharman. 1993, 34, 387-400; Hassan, H.Y.; El Koussi, N.A.; Fargly, Z.S.; J. chem. bulletin 1998, 46, 863-866.). The resin was then washed successively with DMF, methanol and DCM and then dried under reduced pressure. The 2-arylimino-2, 3-dihydrothiazole resin was cleaved under acidic conditions (50% DCM/trifluoroacetic acid) for 1-2 hours and then washed with DCM. The solvent was removed by evaporation, treated under basic conditions (saturated sodium bicarbonate solution), extracted with DCM or eluted with methanol in a basic alumina cartridge (500mg, Interchim) and the free base isolated.

Example 1

N- [3- (2-aminoethyl) -4- (4-chlorophenyl) -1, 3-thiazol-2 (3H) -ylidene ] aniline

A solution of 2-bromo-4' -chloroacetophenone (30.2 mg; 129. mu. mol; 2eq.) in DMF (1ml) was added to the thiourea resin (100 mg; 64.8. mu. mol; load 0.648mmol/g) prepared above. The mixture was stirred at 80 ℃ for 2 hours. The resin was then washed successively with DMF (3X 2ml), methanol (3X 2ml) and DCM (3X 2 ml). The release phase was performed in 1ml of a mixture of 50% DCM/trifluoroacetic acid, stirred for 1h 30 min and after elution with methanol in a basic alumina cartridge (500mg, Interchim) an oil was obtained. The free base was isolated in a quantitative manner (21.3mg) as a yellow oil with a purity of 98% measured by UV spectrophotometry at 220 nm.

NMR ¹H(DMSO D6，100MHz)δ：7.55(s，5H)；7.3(d，2H，J＝7.1Hz)；6.99(d，2H，J＝7.1Hz)；6.21(s，1H，H azole)；3.74(t，2H，J＝6.2Hz，NCH₂)；3.32(broad s，2H，NH₂)；2.72(t，2H，J＝6.2Hz，NCH₂).SM/LC：m/z＝330(M+H)⁺.

Using our automated System (ACTMOS 496), a series of 2-arylimino-2, 3-dihydrothiazoles were synthesized according to method A:

-R1 group:

-R2 group:

-R3 group:

-R4 represents H, alkyl, carbocyclic or heterocyclic aralkyl optionally located on the aryl radical;

Method B

Preparation of urethane Wang resins from aminoalkylanilines

General operation: p-nitrophenylcarbonate Wang resin was treated with excess aminoalkylaniline (5-10eq.) in DCM or DMF and stirred at ambient temperature overnight as described in the literature (Hulme, C.; Peng, J.; Morton, G.; Salvino, J.M.; Herpin, T.; Labaudiniere, R.: tetrahedron letters 1998, 39, 7227-. The resin was washed successively with DMF, methanol and DCM and then dried overnight at 50 ℃ under reduced pressure.

Preparation example 20

4-Aminophenylethylcarbamate Wang resin

A solution of 2- (4-aminophenyl) ethylamine (2.48 g; 17.3 mmol; 5eq.) in 30ml of anhydrous DMF was added to p-nitrophenylcarbonate Wang resin (4.05 g; 3.47 mmol; load 0.857mmol/g) previously swollen in 50ml of anhydrous DMF. The mixture was stirred at ambient temperature overnight and filtered. The resin was washed successively with DMF (10X 30ml), methanol (5X 30ml) and DCM (5X 30 ml). After drying overnight at 50 ℃ under reduced pressure, 3.7g of yellow resin (loaded 0.8mmol/g, calculated from elemental analysis of nitrogen) were isolated and the Kaiser ninhydrin test was positive.

Preparation of thiourea resins from aliphatic isothiocyanates

General operation: aliphatic isothiocyanate (5-10 equivalents) was added to the aminoalkylaniline resin in a solvent such as DCM or DMF and stirred overnight at ambient temperature. After successive washings with DMF and DCM, the thiourea resin was isolated and dried under reduced pressure at 50 ℃ overnight.

Preparation example 21

Wang resin of 4- { [ (phenethylamino) thiocarbonyl ] amino } phenethylcarbamate

10ml of anhydrous DMF and phenethyl isothiocyanate (624. mu.l, 4mmol, 10eq.) were added to the resin (0.5 g; 0.4 mmol; 0.8mmol/g) under argon. The reaction medium is stirred at ambient temperature overnight and the Kaiser ninhydrin test is negative. The resin was then washed successively with DMF (5X 20ml) and DCM (5X 20 ml). Drying at 50 ℃ under reduced pressure gave 488mg of resin at a loading of 0.629mmol/g, as calculated from elemental analysis of sulfur.

Synthesis of 2-arylimino-2, 3-dihydrothiazoles

General operation: the cyclisation stage is carried out between the thiourea resin and the α -bromoketone (2-5 equivalents) in an aprotic solvent, such as dioxane or DMF, at 80 ℃ for 2 hours. The resin was then washed successively with DMF, methanol and DCM and then dried under reduced pressure. The iminothiazole resin was cleaved under acidic conditions (50% DCM/trifluoroacetic acid) for 1-2 hours and then washed with DCM. The solvent was removed by evaporation, treated under basic conditions (saturated sodium bicarbonate solution), extracted with DCM or eluted with methanol in a basic alumina cartridge (500mg, Interchim) and the free base isolated.

Example 2

4- (2-aminoethyl) -N- [4- (4-chlorophenyl) -3-phenethyl-1, 3-thiazolylidene-2 (3H) -yl ] aniline

100mg (62.9. mu. mol, load 0.629mmol/g) of thiourea resin and 2-bromo-4' -chloroacetophenone (30 mg; 125.8. mu. mol; 2eq.) were dissolved in 1ml of DMF and heated to 80 ℃ for 2 hours. The resin was then washed successively with DMF (5X 1ml), methanol (5X 1ml) and DCM (5X 1 ml). The resin was stirred in 1ml of a 50% DCM/trifluoroacetic acid mixture for 1h 30 min at ambient temperature. The resin was washed with DCM (5X 1ml) and the filtrate was evaporated under reduced pressure. The residue was dissolved in methanol and eluted in an alkaline alumina cartridge (500mg, Interchim) in order to quantitatively obtain (27.3mg) the expected product in solid form (UV purity: 97%).

NMR ¹H(DMSO D6，100MHz)δ：7.9(broad s，2H，NH₂)；7.53(d，2H，J＝8.5Hz)；7.32-7.15(m，7H)；7.08-6.9(m，4H)；6.37(s，1H，H azole)；4.07(m，2H，NCH₂)；3.03(m，2H，NCH₂)；2.88(m，4H).MS/LC：m/z＝435(M+H)⁺.

Using our automated System (ACTMOS 496), a series of 2-arylimino-2, 3-dihydrothiazoles were synthesized according to method B:

-R1 group:

-R2 group:

r3 and R4 groups such as those in method A.

Method C

Synthesis of 2-arylimino-1, 3-thiazole-4 (3H) -carboxamides

General operation: regioselective cyclization stages were carried out using α -bromopyruvic acid (2-5eq.) for 2-3 hours at 80 ℃ in an aprotic solvent, such as dioxane or DMF, starting from the thiourea resin prepared in method a. The resin was then washed successively with DMF, methanol and DCM and then dried under reduced pressure. Peptide coupling with different standard coupling agents (4-5eq.) and aminated compounds (4-5eq.) in DMF at ambient temperature for 1-24 hours (Knorr, R.; Trzeciak, A.; Bannwarth, W.; Gillessen, D.; tetrahedron letters 989, 30, 1927-, coupling agents such as Dicyclohexylcarbodiimide (DCC), Diisopropylcarbodiimide (DIC), DIC/N-hydroxybenzotriazole (HOBt) mixtures, benzotriazolyloxytris (dimethylamino) phosphonium hexafluorophosphate (PyBOP), 2- (1H-benzotriazol-1-yl) -1, 1, 3, 3-tetramethyluronium Hexafluorophosphate (HBTU) or 2- (1H-benzotriazol-1-yl) -1, 1, 3, 3-tetramethyluronium tetrafluoroborate (TBTU). The 2-arylimino-1, 3-thiazole-4 (3H) -carboxamide resin was cleaved by treatment under acidic conditions (50% DCM/trifluoroacetic acid) for 1-2 hours and then washed with DCM. The solvent was removed by evaporation, treated under basic conditions (saturated sodium bicarbonate solution), extracted with DCM or eluted with methanol in a basic alumina cartridge (500mg, Interchim) and the free base isolated.

Example 3

3- (4-Aminobutyl) -N-benzhydryl-2- [ (4-bromophenyl) imino]-1, 3-thiazole-4 (3H) -carboxamides

50mg (27.5. mu. mol, load 0.55mmol/g) of the carboxylic acid resin were activated with 14.8mg (0.11mmol, 4eq.) of N-hydroxybenzotriazole and 35.3mg (0.11mmol, 4eq.) of TBTU in 800. mu.l of anhydrous DMF for 15 min. Then 20.7mg (0.11mmol, 4eq.) of aminodiphenylmethane in 200. mu.l anhydrous DMF was added, the resin was filtered after stirring overnight at ambient temperature. The resin was washed sequentially with DMF (5X 1ml), methanol (5X 1ml) and DCM (5X 1ml) and treated under acidic conditions (50% DCM/trifluoroacetic acid) for 1h 30 min. The resin was washed with DCM (5X 1ml) and the filtrate was evaporated under reduced pressure. The residue was dissolved in methanol and eluted in a basic alumina cartridge (500mg, Interchim) to give a pale yellow solid (8.2 mg; yield 55.7%; UV purity at 220nm 94%).

NMR ¹H(DMSO D6，100MHz，δ)：9.6(d；1H；J＝8.6Hz；NH)；7.49(d；2H；J＝8.6Hz)；7.35(s；10H)；6.92(s；1H；H azole)；6.91(d；2H；J＝8.5Hz)；6.27(d；1H；J＝8.5Hz；NHCH)；4.02(m；2H；NCH₂)；3.45(broad m；2H+2H；NH₂ and NCH₂)；1.55-1.24(broad m；4H).MS/LC：m/z＝535(M+H).

Using our automated system (ACTMOS 496), a series of 2-arylimino-1, 3-thiazole-4 (3H) -carboxamides were synthesized according to method C:

the R1 and R2 groups have been as described in method A;

-R3＝-CO-R5；

-R4＝H；

-R5 group:

method D

Synthesis of 2-arylimino-1, 3-thiazole-4 (3H) -carboxamides

General operation: regioselective cyclization stages were carried out using α -bromopyruvic acid (2-5eq.) for 2-3 hours at 80 ℃ in an aprotic solvent, such as dioxane or DMF, starting from the thiourea resin prepared in method B. The resin was then washed successively with DMF, methanol and DCM and then dried under reduced pressure. Peptide coupling with different standard coupling agents (4-5eq.) and aminated compounds (4-5eq.) in DMF at ambient temperature for 1-24 hours (Knorr, R.; Trzeciak, A.; Bannwarth, W.; Gillessen, D.; tetrahedron letters 1989, 30, 1927-, coupling agents such as Dicyclohexylcarbodiimide (DCC), Diisopropylcarbodiimide (DIC), DIC/N-hydroxybenzotriazole (HOBt) mixtures, benzotriazolyloxytris (dimethylamino) phosphonium hexafluorophosphate (PyBOP), 2- (1H-benzotriazol-1-yl) -1, 1, 3, 3-tetramethyluronium Hexafluorophosphate (HBTU) or 2- (1H-benzotriazol-1-yl) -1, 1, 3, 3-tetramethyluronium tetrafluoroborate (TBTU). The 2-arylimino-1, 3-thiazole-4 (3H) -carboxamide resin was cleaved by treatment under acidic conditions (50% DCM/trifluoroacetic acid) for 1-2 hours and then washed with DCM. The solvent was removed by evaporation, treated under basic conditions (saturated sodium bicarbonate solution), extracted with DCM or eluted with methanol in a basic alumina cartridge (500mg, Interchim) and the free base isolated.

Example 4

(2Z) -2- { [4- (2-aminoethyl) phenyl ] imino } -N- (4-chlorobenzyl) -3- (2-phenylethyl) -2, 3-dihydro-1, 3-thiazole-4-carboxamide

A solution of phenethyl isothiocyanate (310 mg; 1.9 mmol; 10eq.) in 3ml of dimethylformamide was added to 200mg (190. mu. mol, loading 0.946mmol/g) of aminated resin (see preparation 20). Stirring overnight at ambient temperature, the Kaiser ninhydrin test was negative. The resin was then washed successively with DMF (5X 3ml) and DCM (5X 3ml) and then dried under vacuum for one hour before adding bromopyruvic acid (63.4 mg; 380. mu. mol; 2eq.) pre-diluted in 3ml of dimethylformamide. The mixture was stirred at 80 ℃ for 2.5 hours. The resin was filtered and washed with DMF (5X 3ml), methanol (3X 3ml) and DCM (5X 3 ml). The carboxylic acid resin was preactivated for 1 hour with 244mg (0.76 mmol; 4eq.) of TBTU diluted in 2ml of anhydrous DMF. Then 110mg (0.76 mmol; 4eq.) of 4-chlorobenzylamine in 1ml of anhydrous DMF was added, the mixture was stirred overnight at ambient temperature and the resin was filtered. The resin was washed sequentially with DMF (5X 3ml), methanol (3X 3ml) and DCM (3X 3ml) and treated under acidic conditions (50% DCM/trifluoroacetic acid) for 1h 30 min. The resin was washed with DCM (5X 1ml) and the filtrate was evaporated under reduced pressure. The residue was dissolved in DCM and neutralized with saturated sodium bicarbonate solution to give a solid after evaporation (38.2 mg; yield 41%; UV purity%).

NMR ¹H(DMSO D6，400MHz，δ)：9.1(m，1H)；7.39(d，2H，J＝8.4Hz)；7.33(d，2H，J＝8.4Hz)；7.25(q，2H，J＝6.8Hz)；7.19(q，1H，J＝7.2Hz)；7.11(m，4H)；6.8(d，2H，J＝8Hz)；6.75(s，1H，H azole)；4.34(d，2H，J＝6Hz)；4.27(t，2H，J＝6.8Hz)；3.14(m，1H)；2.89(t，2H，J＝6.8Hz)；2.73(t，1H，J＝7.2Hz)；2.62(m，2H).MS/LC：m/z＝491.24(M+H)⁺

Using our automated system (ACTMOS 496), a series of 2-arylimino-1, 3-thiazole-4 (3H) -carboxamides were synthesized according to method D:

the R1 and R2 groups have been as described in method B;

-R3＝-CO-R5；

-R4＝H；

the-R5 group has been described in method C.

Method E

Preparation of mono-protected diamine resins functionalized with alpha-bromopropanoic acid

General operation: mono-protected symmetric mono-or di-diamine resins, which have been prepared as described in method a, are functionalized by peptide coupling with alpha-bromopyruvic acid (10eq.), DIC (10eq.), and HOBt (10eq.) in a solvent such as DMF at ambient temperature. After stirring for 2 to 24 hours, the resin was washed successively with DMF and DCM and then dried under vacuum. A negative Kaiser ninhydrin test indicated complete functionalization.

Preparation example 22

2- [ (3-bromo-2-oxopropionyl) amino ] ethyl N-carbamate Wang resin

HOBt (0.93g, 6.88mmol) and alpha-bromopyruvate (1.18g, 6.88mmol) were dissolved in 28ml DMF (0.5M). DIC (1.07 ml; 6.88mmol) was then added via syringe to activate the acid. The mixture was stirred at ambient temperature for about 15 minutes and then added to ethylenediamine N-carbamate Wang resin (0.8 g; 0.688 mmol; loading 0.86 mmol/g). After stirring at ambient temperature for 3 hours, the Kaiser ninhydrin test was negative, the resin was filtered, washed successively with DMF (5X 20ml) and DCM (5X 20ml) and then dried under vacuum. Ochre resin (0.812g) was obtained with a loading of 0.525mmol/g, calculated from elemental analysis of bromine.

Synthesis of 2-arylimino-1, 3-thiazole-4 (3H) -carboxamides

General operation: thiourea formation is carried out by mixing equimolar amounts of a primary amine with an aromatic or heteroaromatic isothiocyanate in a solvent such as DMF or dioxane. After stirring at ambient temperature for 2 to 24 hours, thiourea (2-5eq.) was added to the functionalized resin, followed by heating at 80 ℃ for 2 to 4 hours. The 2-arylimino-1, 3-thiazole-4 (3H) -carboxamide resin was cleaved by treatment under acidic conditions (50% DCM/trifluoroacetic acid) for 1-2 hours and then washed with DCM. The solvent was removed by evaporation, treated under basic conditions (saturated sodium bicarbonate solution), extracted with DCM or eluted with methanol in a basic alumina cartridge (500mg, Interchim) and the free base isolated.

Example 5

(2Z) -N- (2-aminoethyl) -3- [2- (3, 4-dimethoxyphenyl) ethyl ] -2- (phenylimino) -2, 3-dihydro-1, 3-thiazole-4-carboxamide

Mu.l (105. mu. mol; 2eq.) of beta- (3, 4-dimethoxyphenyl) ethylamine and 12.6. mu.l (105. mu. mol; 2eq.) of phenylisothiocyanate were stirred in 1ml of DMF for 18 hours. Thiourea was added to 100mg (52.5. mu. mol; loading 0.525mmol/g) of the resin (preparation 22), and the mixture was heated at 80 ℃ for 3 hours. The resin was then filtered and washed successively with DMF (5X 1ml), methanol (5X 1ml) and DCM (5X 1 ml). The resin was dried under vacuum and then 1ml of 50% DCM/TFA mixture was added. Stirring was carried out at ambient temperature for 1.5 h, the resin was filtered and washed with DCM. The residue recovered after evaporation was eluted with methanol in a basic alumina column in order to isolate 22.2mg (quantitative yield; UV purity at 230nm 93.4%) of a brown solid, corresponding to the free amine.

NMR ¹H(DMSO D6，100MHz，δ)：8.42(m，1H，NH)；7.32(t，2H，J＝7.1Hz)；7.08-6.63(m，6H)；5.76(s，1H，H azole)；4.31(t，2H，J＝6.6Hz)；3.72(s，6H，OCH₃)；3.32(broad s，2H)；3.17(m，2H)；2.89(m，2H)；2.62(m，2H).MS/LC：m/z＝427.17(M+H)⁺.

Using our automated System (ACTMOS 496), a series of 2-arylimino-1, 3-thiazole-4 (3H) -carboxamides were synthesized according to method E:

-R1 group:

the R2 group has been described as in method A;

-R3＝-CO-R5；

-R4＝H；

-R5 group:

method F

Preparation of Mono-protected diamine resins functionalized with N-protected amino acids (Fmoc)

General operation: mono-protected diamine resins were coupled with a peptide of commercially available (Bunin, B.A. "Combined index" Academic Press, 1998, p.77-82) N-Fmoc amino acids (4-10eq.) in DMF at ambient temperature for 1 to 24 hours using different standard coupling agents (4-10eq.), coupling agents such as Dicyclohexylcarbodiimide (DCC), Diisopropylcarbodiimide (DIC), DIC/N-hydroxybenzotriazole (HOBt) mixtures, benzotriazolyloxytris (dimethylamino) phosphonium hexafluorophosphate (PyBOP), 2- (1H-benzotriazol-1-yl) -1, 1, 3, 3-tetramethyluronium Hexafluorophosphate (HBTU) or 2- (1H-benzotriazol-1-yl) -1, 1, 3, 3-tetramethyluronium tetrafluoroborate (TBTU). The resin was then washed successively with DMF and DCM. The coupling procedure can be repeated one to two times until the Kaiser ninhydrin test is negative.

Preparation example 23

4- [ ({ [ (9H-fluoren-9-ylmethoxy) carbonyl ] amino } acetyl) amino ] butyl N-carbamate Wang resin

Fmoc-Gly-OH acid (2.36g, 7.94mmol) was activated with HOBt (1.07g, 7.94mmol) and DIC (1.25ml, 7.94mmol) in 22ml DMF for 5 min before the mixture was added to butylamine N-carbamate Wang resin (1g, loading 0.794mmol/g) previously swollen in 10ml DMF. After stirring at ambient temperature for 18 h, the resin was washed successively with DMF (5X 20ml) and DCM (5X 20ml) and then dried under vacuum. This gives 1.27g of a pale yellow resin which is negative in the Kaiser ninhydrin test.

Preparation of thiourea resins

General operation: the resin was deprotected with 20% DMF/piperidine mixture. After stirring at ambient temperature for one hour, the resin was filtered and washed successively with DMF and DCM. The deprotection/washing procedure was repeated a second time and the resin was dried under vacuum. The latter is pre-swollen in a solvent such as DMF or DCM, then the aromatic or heteroaromatic isothiocyanate (5-10eq.) is added. The mixture was stirred at ambient temperature for 2 to 24 hours, then the resin was filtered and washed successively with DMF and DCM. The resin was then dried under vacuum and a negative Kaiser ninhydrin test confirmed that the substitution reaction was complete.

Preparation example 24

4- [ ({ [ (1-naphthylamino) thiocarbonyl ] amino } acetyl) amino ] butyl N-carbamate Wang resin

1.27g of the above resin (cf. preparation 23) were deprotected with 14ml of 20% DMF/piperidine. The mixture was stirred at ambient temperature for one hour. The resin was then filtered and washed with DMF (5X 30ml) and DCM (5X 30 ml). The deprotection/washing procedure was repeated once and then the resin was dried under vacuum. This gave 0.781g of a pale yellow resin, at a loading of 0.758mmol/g, as calculated from elemental analysis of sulfur. 416mg (2.2mmol, 10eq.) of 1-naphthyl isothiocyanate was diluted in 6ml of DMF and added to 0.3g (0.22mmol) of the thiourea resin. The mixture was stirred at ambient temperature for 18 hours. The resin was filtered and then washed successively with DMF (5X 20ml) and DCM (5X 20 ml). After drying under vacuum 310mg of pale yellow resin were isolated with a loading of 0.66mmol/g, calculated from elemental analysis of nitrogen.

Synthesis of 2-arylimino-2, 3-dihydrothiazoles

General operation: regioselective cyclization stages between thiourea resin and α -bromoketone (2-5eq.) were carried out at 80 ℃ for 2 to 3 hours in aprotic solvents such as dioxane, DMF or N-methylpyrrolidinone. The resin was then washed successively with DMF, methanol and DCM and then dried under reduced pressure. The 2-arylimino-2, 3-dihydrothiazole resin was cleaved under acidic conditions (50% DCM/trifluoroacetic acid) for 1 to 2 hours, then washed with DCM. The solvent was removed by evaporation, treated under basic conditions (saturated sodium bicarbonate solution), extracted with DCM or eluted with methanol in a basic alumina cartridge (500mg, Interchim) and the free base isolated.

Example 6

N- (4-aminobutyl) -2- ((2Z) -4- (4-chlorophenyl) -2- (1-naphthylimino) -1, 3-thiazol-3 (2H) -yl) acetamide

80mg (52.8. mu. mol, loading 0.66mmol/g) of thiourea resin (preparation 24) and 25.1mg (105.6mmol, 2eq.) of 2-bromo-4' -chloroacetophenone were diluted in 1ml of DMF. The mixture was heated at 80 ℃ for 2 hours. The resin was filtered and then washed with DMF (5X 1ml), methanol (5X 1ml) and DCM (5X 1ml) and dried under vacuum. 1ml of 50% DCM/TFA mixture was added, followed by stirring for 1 hour 30 minutes. The resin was filtered and washed with DCM. The filtrate was evaporated and then diluted again in methanol eluting on basic alumina. After evaporation, 20.6mg (yield 84%; UV purity at 220nm 94.2%) of a yellow solid are thus obtained, corresponding to the free base.

NMR ¹H(DMSO D6，100MHz，δ)：8.36(t，1H，J＝4.7Hz，NH）；8.12(dd，1H，J＝2.1and 7.3Hz)；7.87(dd，1H，J＝2.7and 6.3Hz)；7.63-7.34(m，8H)；7.13(dd，1H，J＝1.6and 6.7Hz)；6.33(s，1H，H azole)；4.44(broad s，2H)；3.14(m，2H)；2.7(m，2H)；1.5(m，4H).MS/LC：m/z＝465.21(M+H)⁺.

Using our automated System (ACTMOS 496), a series of 2-arylimino-2, 3-thiazoles were synthesized according to method F:

-R1＝-C(R11R12)-CO-R10；

the R2, R3 and R4 groups have been as described in method A;

-R10 group:

-R11＝H；

-R12 group:

examples

Examples obtained according to methods A, B, C, D, E and F described above are shown below. These examples are provided only to illustrate the above-described operations and are in no way to be construed as limiting the scope of the invention.

The retention time (rt) and mass spectrum (M + H) of the resulting compounds have been identified⁺。

Pharmacological Properties of the products of the invention

The compounds of the present invention can and have been tested for their affinity for different subtypes of somatostatin receptors according to the following procedures.

Affinity studies for human somatostatin receptor subtypes:

by measuring the length of the pair of [2 ]¹²⁵I-Tyr¹¹]Inhibition of binding of SRIF-14 to transfected CHO-K1 Compounds of the invention were assayed against subtypes 1 to 5 of human somatostatin receptors (sst, respectively)₁、sst₂、sst₃、sst₄And sst₅) The affinity of (a).

Human growth hormone release inhibitory factor sst has been cloned₁A genomic fragment of a receptor gene. The 1.5Kb PstI-XmnI segment containing 100bp of the non-transcribed 5 'region, the entire 1.17Kb coding region and 230bp of the non-transcribed 3' region was modified by the addition of linker Bg1 II. The resulting DNA fragment was subcloned at the BamHI site of pCMV-81 in order to prepare an expression plasmid (supplied by dr. Expression of sst in a Stable manner by transfection in CHO-K1 cells (ATCC) using calcium phosphate coprecipitation₁A clonal cell line of recipients. Plasmid pRSV-neo (ATCC) was included as a selection marker. Clonal cell lines were selected in RPMI 1640 medium containing 0.5mg/ml G418(Gibco) and then subjected to cyclic cloning and proliferation in culture.

Human growth hormone release inhibitory factor sst₂The receptor gene was supplied by Dr.G.Bell (Univ.Chicago), isolated as a 1.7Kb BamHI-HindIII DNA genomic fragment and subcloned in the plasmid vector pGEM3Z (Promega). A1.7 Kb BamH1-HindII fragment was inserted into the endonuclease restriction site of plasmid pCMV5 to construct an expression vector for mammalian cells. A clonal cell line was obtained by transfection in CHO-K1 cells using calcium phosphate co-precipitation. Plasmid pRSV-neo was included as a selection marker.

Separating sst₃The genomic fragment of the receptor contained the entire coding sequence in the 2.4Kb BamHI/HindIII fragment. After terminal modification and addition of EcoR1 linker, a 2.0Kb NcoI-HindIII fragment was inserted at EcoR1 site of vector pCMV to construct expression plasmid pCMV-h3 in mammals. Expression of sst in a Stable manner by transfection in CHO-K1 cells (ATCC) using calcium phosphate coprecipitation₃A clonal cell line of recipients. Plasmid pRSV-neo (ATCC) was included as a selection marker. Clonal cell lines were selected in RPMI 1640 medium containing 0.5mg/ml G418(Gibco) and then subjected to cyclic cloning and proliferation in culture.

Human sst₄The expression plasmid for the receptor, pCMV-HX, was supplied by Dr.Graeme Bell (Univ.Chicago). The vector contains a human sst encoding 1.4Kb NheI-NheI, 456pb non-transcribed 5 'region and 200pb non-transcribed 3' region₄The genomic fragment of the receptor was cloned into pCMV-HX at the XbaI/EcoR1 site. Expression of sst in a stable manner was obtained by transfection in CHO-K1 cells (ATCC) using calcium phosphate co-precipitation₄A clonal cell line of recipients. Plasmid pRSV-neo (ATCC) was included as a selection marker. Clonal cell lines were selected in RPMI 1640 medium containing 0.5mg/ml G418(Gibco) and then subjected to cyclic cloning and proliferation in culture.

Using genomic lambda clones as probes, human sst-equivalent was obtained by PCR₅A receptor gene provided by dr. The resulting 1.2KbPCR fragment contained a 21 base pair non-transcribed 5 'region, the entire coding region and a 55pb non-transcribed 3' region. The clone was inserted at EcoR1 site of plasmid pBSSK (+). The insert was recovered as a 1.2Kb HindHI-XbaI fragment and used for subcloning into the mammalian expression vector pCVM 5. Expression of sst in a Stable manner by transfection in CHO-K1 cells (ATCC) using calcium phosphate coprecipitation₅A clonal cell line of recipients. Plasmid pRSV-neo (ATCC) was included as a selection marker. Clonal cell lines were selected in RPMI 1640 medium containing 0.5mg/ml G418(Gibco) and then subjected to cyclic cloning and proliferation in culture.

CHO-K1 cells expressing human sst receptors in a stable manner were cultured in RPMI 1640 medium containing 10% fetal bovine serum and 0.4mg/ml geneticin. Cells were harvested with 0.5mM EDTA and centrifuged at 500g for about 5 minutes at about 4 ℃. The pellet was resuspended in Tris 50mM, pH 7.4 and centrifuged twice at 500g for approximately 5 minutes at approximately 4 ℃. Cells were lysed by sonication and then centrifuged at 39000g for approximately 10 minutes at 4 ℃. The sediment was resuspended in the same buffer, centrifuged at 50000g for about 10 minutes at about 4 ℃ and the cell membranes in the resulting sediment were stored at-80 ℃.

The reaction is carried out in a 96-well polypropylene plate¹²⁵I-Tyr¹¹]Competitive inhibition of SRIF-14 binding assay, in duplicate. Containing 0.2% BSA and MgCl₂5mM, aprotinin 200KIU/ml, bacitracin 0.02mg/ml, phenylmethylsulfonyl fluoride 0.02mg/ml in HEPES50mM buffer (pH 7.4), cell membrane (10. mu.g protein/well) was bound at about 37 ℃¹²⁵I-Tyr¹¹]SRIF-14(0.05nM) was incubated for approximately 60 minutes.

Using Filtermate 196(Packard), free [ sic ] was immediately filtered through GF/C glass fiber filter plates (Unifilter, Packard) previously impregnated with 0.1% polyethylenimine (P.E.I. ])¹²⁵I-Tyr¹¹]SRIF-14 and bound [ alpha ], [ alpha ]¹²⁵I-Tyr¹¹]And (5) separating the SRIF-14. The filter plates were washed with 50mM HEPES buffer at about 0-4 ℃ for about 4 seconds and their radioactivity was measured using a counter (Packardtop Count).

Specific binding was obtained by subtracting non-specific binding (measured in the presence of 0.1. mu.M SRIF-14) from total binding. Analysis of the data on binding by computer-assisted non-linear regression analysis (MDL) to determine the inhibition constant (K)₁) The value of (c).

The following assays were used to determine agonist or antagonist properties of the compounds of the invention.

And (4) functional test: inhibition of intracellular cAMP production

CHO-K1 cells expressing human growth hormone Release inhibitor receptor subtype (SRIF-14) were cultured in RPMI 1640 medium containing 10% fetal bovine serum and 0.4mg/ml geneticin 24-well plates. The culture medium was changed the day before the experiment.

At a ratio of 10⁵The cells per well were washed twice with 0.5ml of fresh RPMI medium containing 0.2% BSA, quenched with 0.5mM 3-isobutyl-1-methylxanthine (IBMX), and incubated at about 37 ℃ for about 5 minutes.

Cyclic AMP production was stimulated by the addition of 1mM Forskolin (FSK) for 15-30 minutes at approximately 37 deg.C.

Simultaneously adding FSK (1 μ M) and SRIF-14 (10)^-12M to 10^-6) And a test compound (10)^-10M to 10^-5M), measuring the inhibitory effect of the agonist compound on growth hormone release inhibitor.

FSK (1. mu.M), SRIF-14(1 to 10nM) and test compound (10 nM) were added simultaneously^-10M to 10^-5M), measuring the antagonistic effect of the compound.

The reaction medium is drained and 200ml of 0.1N HCl are added. The amount of cAMP was measured by radioimmunoassay (FlashPlate SMP001A kit, New England Nuclear).

As a result:

tests carried out according to the above-described protocol have demonstrated that the products of general formula (I) defined in the present application have a good affinity for at least one subtype of somatostatin receptor, the inhibition constant K being the product shown in the table below for certain exemplary compounds, precisely₁Less than micromolar.

Claims

1. A compound of the general formula (I):

wherein:

r1 representsA radical, wherein n is 1-6;

r2 representsA radical;

r3 represents CO-R5;

r4 represents H;

r5 represents one of the following radicals:

or a salt of a compound of formula (I).

2. A compound of the formula

Or a salt of said compound.

3. A pharmaceutical composition comprising a compound according to one of claims 1 or 2 or a pharmaceutically acceptable salt of said compound as an active ingredient.

4. Use of a compound according to one of claims 1 or 2 or of a pharmaceutically acceptable salt of said compound for the preparation of a medicament intended for the treatment of a pathological state or disease in which one or more somatostatin receptors are involved.

5. Use according to claim 4, characterized in that the pathological state or disease to be treated is selected from the group comprising: acromegaly, pituitary adenoma, Cushing's disease, gonadotropinoma and prolactinoma, glucocorticoid catabolic side effects, insulin-dependent diabetes mellitus, diabetic retinopathy, diabetic nephropathy, syndrome X, dawn phenomenon, vascular disease, angioplasty, hyperthyroidism, gigantism, endocrine gastroenteropancreatic tumors, tumor of growth hormone releasing factor of esophageal varices and acute hemorrhage, ulcer, gastroesophageal reflux, gastroduodenal reflux, pancreatitis, enterocutaneous and pancreatic fistula with diarrhea, refractory diarrhea of acquired immunodeficiency syndrome, chronic secretory diarrhea, diarrhea associated with irritable bowel syndrome, diarrhea induced by chemotherapy, disorders associated with gastrin-releasing peptide, secondary lesions of intestinal transplantation, portal hypertension and variceal hemorrhage of cirrhosis patients, gastrointestinal bleeding, Gastroduodenal ulcer bleeding, graft vascular bleeding, crohn's disease, systemic sclerosis, dumping syndrome, small bowel syndrome, hypotension, scleroderma and medullary thyroid carcinoma, diseases associated with cellular hyperproliferation, fibrosis, headache, pain, inflammation, panic attacks, chemotherapy, wound healing, renal insufficiency due to developmental delay, hyperlipidemia, obesity and developmental delay associated with obesity, uterine developmental delay, skeletal dysplasia, noonan syndrome, sleep apnea syndrome, graves ' disease, ovarian polycystic tumor disease, pancreatic pseudocysts and ascites, leukemia, meningioma, cancer cachexia, inhibition of helicobacter pylori, psoriasis, chronic allograft rejection, and alzheimer's disease and osteoporosis.

6. The use of claim 5, wherein said endocrine gastroenteropancreatic tumor is selected from the group consisting of carcinoid syndrome, pancreatic tumors, insulinoma, gastrinoma, and Zollinger-Ellison syndrome.

7. The use of claim 5, wherein the disease associated with cellular hyperproliferation is cancer.

8. The use of claim 7, wherein the cancer is selected from the group consisting of breast cancer, prostate cancer, thyroid cancer, and pancreatic and colorectal cancers.

9. The use of claim 5, wherein said fibrosis is selected from the group consisting of renal fibrosis, liver fibrosis, lung fibrosis, skin fibrosis, central nervous system fibrosis and nasal fibrosis and fibrosis induced by chemotherapy.

10. The use of claim 5, wherein said headache is a headache associated with pituitary tumors.

11. The use of claim 5, wherein the inflammation is arthritis.

12. Use according to claim 5, characterized in that the pathological conditions or diseases to be treated are selected from the group comprising: acromegaly, pituitary adenoma, or endocrine gastroenteropancreatic tumors, and gastrointestinal bleeding.

13. The use according to claim 12, wherein said pituitary adenoma or endocrine gastroenteropancreatic neoplasm is carcinoid syndrome.