MXPA01011267A - 1,5-benzodiazepine derivatives. - Google Patents

Abstract

An enantiomerically enriched compound of Formula (I) is disclosed, processes for its preparation, pharmaceutical compositions containing it and the use therefore, for the treatment of CCK-A mediated diseases or conditions, such as obesity.

Description

1, 5-BENZODIAZEPINE DERIVATIVES FIELD OF THE INVENTION This invention relates to novel derivatives of 5, 5-benzodiazepine, to processes for their preparation, to pharmaceutical compositions containing them and to their use in medicine. More particularly, it refers to compounds that show an agonist activity for the CCK-A receptors.

BACKGROUND OF THE INVENTION Cholecystokinin (CCK) is a peptide found in the gastrointestinal tract and in the central nervous system, see A.J. Prange et al., Ann. Reports Med. Chem. 17, 31, 33 (1982), J.A. Williams, Biomed Res. 3, 107 (1982) and V. Mutt, Gastrointestinal Hormones, G.B.J. Green, Ed., Raven Press, NY, 169. CCK has been implicated inter alia as a physiological satiety hormone involved in the regulation of appetite, see Della-Ferra et al, Science, 206, 471 (1979), Saito et al. contributors, Na ture, 289, 599, (1981), GP Smith, Ea ting and Its Disorders, A.J. Stun ard and E. Stellar, Eds, Raven Press, New York, 67 (1984), as a regulator of gallbladder contraction and secretion of pancreatic enzymes, an inhibitor of gastric emptying, and as a neurotransmitter, see AJ Prange, supra, J.A. REF. 1 33987 - & amp;, * l ^ j ^^^ tg & ^^^^^^ & fcb ^^^^? ^ ¿^ ^ m Ü¡Ít¡¡¡ Williams, Biomed Res. , 3, 107 (1982), J.E. Morley, Life Sci. 30, 479, (1982). Gastrin is a peptide involved in the secretion of gastric acid and pepsin in the stomach, see 1. Sandvick et al., American J. Physiology, 260, G295 (1991), C.W. Lin and collaborators, American J.

Physiology, 262, G1113, (1992). CCK and gastrin share structural homology in their C-terminal tetrapeptide: Trp-Met-Asp-Phe. Two subtypes of CCK receptors, designated CCK-A and CCK-B, have been identified, and both have been found in the central and peripheral nervous systems. Recently it has been reported that the CCK-B receptors are similar to the gastrin receptor, see Pisegna, J.R., of Weerth, A, Huppi, K, Wank, S.A., Biochem. Bi ophys. Res. Commun. 189, 296-303 (1992). CCK-A receptors are predominantly located in peripheral tissues that include the pancreas, gallbladder, ileum, pyloric sphincter, and vagal afferent nerve fibers; CCK-A receptors are found to a lesser degree in the brain, see T.H. Moran et al., Brain Res. , 362, 175-179 (1986), D.R. Hill et al., Brain Res, 4545, 101, (1988), D.R. Hill et al., Neurosci Let t. , 89, 133, (1988), R.W. Barret et al., Mol. Pharmacol. , 36, 285, (1989), D.R. Hill et al., J. Neurosci, 10, 1070 (1990), V. Daugue et al., Pharmacol Biochem Behac, 33, 637, (1989), whereas CCK-B receptors are found predominantly in the brain, see V.J. Lotti and R.S.L Chang, Proc. Natl Acad. Sci. U. S A., 83, 4923 (1986), J.N. Crawley, Trends Pharm. Sci. , 88, 232, (1991). 5 The agonist activity of the CCK has been associated with the inhibition of the ingestion of food in animals, and in this way, with its weight loss, see Della-Fera et al., Supra, K.E. Asin and collaborators, Intl. Conference on Obesi ty, abstract pp.40 (1990). It has been suggested 10 that CCK acts in the periphery through the vagal fibers and not directly in the brain, to produce satiety, see Smith, G.P. and Cushin, B.J., Neurosci ence Abstr. , 4, 180 (1978), Smith, G.P., Jerome, C, Cushin, B.J., Eternal, R., and Simansky, K.J., Sci ence, 212, 687-689 15 (1981). U.S. Patent No. 5,646,140 (Sugg et al.) Discloses certain 3-amino-1,5-benzodiazepine compounds showing agonist activity by the CCK-A receptor, thereby allowing them to modulate the hormones 20 gastrin and cholecystokinin (CCK) in mammals. See in particular the compound of Example 7. Certain of these compounds also exhibit antagonist activity at the CCK-B receptors.

DESCRIPTION OF THE INVENTION Briefly, in one aspect, the present invention provides an enantiomerically enriched compound, of Formula (I), or a pharmaceutically acceptable salt or solvate thereof.

(I) The compound of Formula (I) is 3- acid. { 3- [l- (isopropyl-phenyl-carbamoylmethyl) -2,4-dioxo-5-phenyl-2, 3, 4, 5- 20 tetrahydro-lH-benzo [b] [1,4] diazepin-3-yl] ureido} benzo? co. This compound has a chiral carbon in the benzodiazepine ring. Applicants have found that the enantiomer that rotates light in the positive direction is preferred, under the conditions described 25 later. This enantiomer, which is hereinafter referred to as the (+) enantiomer, has the configuration (S), according to the Cahn Ingold Prelog convention. Applicants have found that this isomer has improved properties with respect to the racemic mixture and is therefore more convenient than the racemic mixture for the treatment of obesity and other diseases or conditions mediated by CCK-A. As used herein, "enantiomerically enriched" means that there is more of the (+) enantiomer than of the (-) enantiomer, which is the opposite of the case of the racemic mixture, which has equal amounts of each isomer. As used herein "the compound of this invention" or "the enantiomerically enriched compound of this invention, and expressions containing these phrases or similar phrases, include the pharmaceutically acceptable salts and solvates thereof." The "(+) enantiomer" refers to the optical rotation of the enantiomer and not the salts and solvates thereof The preferred salts and solvates will be salts and solvates of the (+) - enantiomer of the compound of Formula (I) regardless of the optical rotation of the salt or solvate. Preferably, the (+) enantiomer is at least 90% of the total amount of the enantiomerically enriched compound. More preferably, the (+) enantiomer is at least 96% of the total amount of the compound. Most preferably, the (+) enantiomer is at least 99% of the total amount of the compound.

The enantiomer (+) of the present invention shows a CCKA agonist activity and can be considered as a total agonist of cholecystokinin, because it binds to the CCK-A receptors and totally stimulates the contraction of the gall bladder and reduces the feeding in animal models. For example, the (+) enantiomer of this invention should be useful for the treatment of obesity, as well as related pathologies, such as hypertension, gall bladder stasis, and diabetes, indirectly through weight loss, and directly through delayed gastric emptying, mediated by CCK-A. In addition, the (+) enantiomer described herein provides novel approaches to induce satiety, to provide appetite regulation and to modify food intake in mammals, especially humans, to regulate appetite, treat obesity and maintain loss of appetite. weight. Therefore, in a further aspect of the present invention, there is provided herein a method for the treatment, in a mammal, including man, of a disease or condition mediated by CCK-A, which comprises administering to a patient a therapeutically effective amount of the (+) enantiomer of this invention. & In accordance with another aspect, the present invention provides the use of an enantiomerically enriched compound of this invention, or a pharmaceutically acceptable salt or solvate thereof, for the manufacture of a medicament for the treatment of diseases or conditions mediated by the CCK. -TO. Those skilled in the art will appreciate that when treatment is referred to herein, it extends to prophylaxis as well as treatment of established diseases or symptoms. In addition, it will be appreciated that the amount of the preferred enantiomer of the invention required for use in the treatment, it will vary with the nature of the condition being treated and with the age and condition of the patient, and finally it will be at the decision of the doctor or veterinarian who attends the case. However, in general, the doses employed for the treatment of adult humans will commonly be in the range of 0.02 to 5000 mg per day, for example, from 1 to 1500 mg per day. The desired dose may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, for example, in two, three, four or more secondary doses per day. Although it is possible that the enantiomerically enriched compound of the present invention is therapeutically administered as the pure chemical, It is preferable to present the active ingredient as a pharmaceutical composition. Accordingly, the present invention further provides a pharmaceutical composition comprising the enantiomerically enriched compound, of this invention, together with one or more pharmaceutically acceptable carriers and / or excipients therefor and, optionally, other therapeutic and / or prophylactic ingredients. The carriers and / or excipients therefor must be "acceptable" in the sense that they are compatible with the other ingredients of the formulation and that they are not harmful to the one receiving the same. Formulations of the present invention include those formulated especially for oral, buccal, parenteral, implant, topical or rectal administration, however oral administration is preferred. For buccal administration, the composition may take the form of tablets or lozenges formulated in a conventional manner. Tablets and capsules for oral administration may contain conventional excipients such as binding agents (eg, syrup, acacia, gelatin, sorbitol, tragacanth, starch mucilage or polyvinylpyrrolidone), fillers (eg, lactose, sugar, cellulose) microcrystalline, corn starch, calcium phosphate or sorbitol), lubricants (eg, magnesium stearate, stearic acid, talc, polyethylene glycol or silica), disintegrants (e.g., potato starch or sodium starch glycolate) or wetting agents, such as sodium lauryl sulfate. The tablets may be coated according to methods well known in the art, including enteric coatings. Alternatively, the preferred enantiomer of the present invention can be incorporated into oral liquid preparations such as, for example, aqueous or oily suspensions, solutions, emulsions, syrups or elixirs. In addition, formulations containing this preferred enantiomer may be presented as a dry product for constitution with water or other suitable vehicle before use. These liquid preparations may contain conventional additives such as suspending agents, such as, for example, sorbitol syrup, methylcellulose, glucose / sugar syrup, gelatin, hydroxyethylcellulose, carboxymethylcellulose, aluminum stearate gel or hydrogenated edible fats; emulsifying agents such as lecithin, sorbitan mono-oleate or acacia; non-aqueous vehicles (which may include edible oils) such as almond oil, fractionated coconut oil, oily esters, propylene glycol or ethyl alcohol; and preservatives such as methyl or propyl p-hydroxybenzoates, or ascorbic acid. These preparations are They can also be formulated as suppositories, for example, containing conventional suppository bases, such as cocoa butter or other glycerides, In addition, the compositions of the present invention can be formulated for parenteral administration by injection or by injection. Continuous infusion: Formulations for injection may take such forms as suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain agents for 10 formulations, as suspending, stabilizing and / or dispersing agents. Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle (eg, sterile, pyrogen-free water) before use. The composition according to the invention can also be formulated as a deposit preparation. These long-acting formulations can be administered by implant (e.g., subcutaneously or intramuscularly) or by intramuscular injection.

Accordingly, the preferred enantiomer of the invention can be formulated with suitable polymeric or hydrophobic materials (for example, as an emulsion in an acceptable oil), ion exchange resins or as sparingly soluble derivatives, such as, for example, a salt poorly 25 soluble. - ^^^ móí ^^^^^^^^^ * ^ ¿dfe * The compositions according to the invention may contain between 0.1 and 99% of the active ingredient, conveniently 30 to 95% for tablets and capsules, and 3 to 50% for liquid preparations. The (+) enantiomer of this invention can be produced by first making the racemic mixture, as described in Example 7 of the US Patent No. 5,646,140 and then separating the enantiomers by chiral chromatography. Alternatively the (+) enantiomer can be prepared by the reaction of the appropriate enantiomer, especially the (S) -enantiomer of the amine of the formula di); 25 with an isocyanate of formula (III, wherein R is a carboxyl protective group, eg, t-butyl), an imidazolide of formula (IV, wherein R is a carboxyl protective group, eg, t-butyl) or an optionally substituted phenyl carbonate of formula (V; wherein R is a carboxyl protecting group, eg, t-butyl and Ri is hydrogen or a conventional phenyl substituent, eg, N02) followed by removal of the carboxyl protecting group R.

(V) The reaction conveniently takes place in the presence of a suitable solvent such as an ether (e.g., tetrahydrofuran) or a halogenated hydrocarbon (e.g., dichloromethane) or nitrile (e.g., acetonitrile) ^ ^ fa ^^ 1 at a temperature that is in the range of 0 to 80 ° C. Conveniently the required enantiomer of the amine (II) can be used in the form of a salt thereof, for example, a salt of the R-camforsulfonic acid and in this embodiment the reaction can be carried out in the presence of a base, example, of a tertiary amine such as triethylamine. The hydrolysis of the carboxyl protecting group can be carried out using conventional procedures.

(Protecting groups in Organic Synthesis T. Greene, Ed., Wiley Interscience, New York, p-168, 1981). Thus, for example, when R is a t-butyl group, it can be removed by hydrolysis with an appropriate acid such as hydrochloric acid, trifluoroacetic acid or formic acid, using established procedures. For example, by reacting with hydrochloric acid in a solvent such as 1,4-dioxane, or by reacting with formic acid in a solvent such as acetone or aqueous acetone and with heating. The required (S) enantiomer of the amine of Formula (II); it can be prepared by separating the corresponding racemic amine through chiral HPLC chromatography, or by means of asymmetric separation induced by crystallization via the R-camforsulfonic acid salt. The racemic amine (II) can be prepared by the method described in Intermediate Compound 11 of U.S. Patent No. 5,646,140. Alternatively, the racemic amine (II) can be prepared by concomitant reduction and hydrogenolysis of the oxime (VI) wherein R2 is an optionally substituted benzyl group.

--"*•* * * (SAW) The reaction is conveniently carried out using a suitable palladium catalyst, for example palladium on carbon, for example 5% Pd on carbon in the presence of hydrogen or aqueous ammonium formate, and in a solvent such as an aqueous alkanol, example ethanol, isopropanol or methylated, industrial, or tetrahydrofuran volatile compounds. Conveniently the reaction is carried out with heating, for example from 40 to 80 ° such as 60 ° C. Examples of suitable R2 groups, for use in the reaction, include benzyl, or substituted benzyl, such as p-methoxybenzyl, or benzhydryl. The oxime (VI) can be prepared by the reaction of the ortho-phenylenediamine derivative (VII). ^? m ?? ¡t H ^ i ^^^^^^ J with an activated derivative of the diacid (VIII) wherein R2 is an optionally substituted benzyl group.

Conveniently the activated derivative of the diacid (VIII) is the corresponding diacyl halide, for example the chloride, and this is prepared in situ by the reaction of the diacid (VIII) with an oxalyl halide, for example oxalyl chloride. The reaction is conveniently carried out in an aprotic solvent, such as an ester, for example ethyl acetate, toluene, dichloromethane, dimethoxyether or mixtures thereof, and in the presence of dimethylformamide.

The diacid (VIII) is conveniently prepared by the reaction of a dialkyl ketomalonate, for example a diethyl ketomalonate with the corresponding hydroxylamine R2ONH2 in a solvent such as an alkanol, for example methanol or industrial methylated volatile compounds and in the presence of a base, for example pyridine, followed by hydrolysis of the corresponding dialkyl-oximino malonate using aqueous sodium hydroxide. In a further aspect, the invention provides a process for preparing the (S) -enantiomer of the compound of formula (I) substantially free of the (R) -enantiomer thereof from the racemic amine (II) as described above, wherein the Racemic amine (II) has been prepared from the oxime (VI) as described above and more particularly, wherein the oxime (VI) has been prepared from the compounds (VII) and (VIII). The isocyanates of Formula (III) can be purchased or prepared by reacting the corresponding amine (VI) with phosgene or triphosgene in a suitable solvent such as methylene chloride. The imidazolides of Formula (IV) can be prepared by treating the corresponding amine (VI) with carbonyldiimidazole in a suitable solvent (dichloromethane, ether, tetrahydrofuran) at a temperature ranging from 0 to 80 ° C (conveniently at room temperature) . The optionally substituted phenyl carbamates of Formula (V) can be prepared by reaction of the corresponding amine (VI) with the optionally substituted phenyl chloroformate in the presence of a base (pyridine, triethylamine) in a suitable solvent (dichloromethane) and a temperature from 0 to 50 ° C. The amines of the formula (VI) are already known compounds and can be prepared by methods analogous to those used to prepare the known compounds. The following examples, which are not limiting, illustrate the invention. In the examples, the abbreviations have the following meanings, EtOAc = ethyl acetate; MeOH = methanol, DMF = N, N-dimethylformamide; IPA = isopropyl alcohol; IMS = industrial methylated volatile compounds.

Intermediate Compound 1 (+) -2- (3-Amino-2,4-dioxo-5-phenyl-2,3,4,5-tetrahydrobenzo- [b] [1,4] diazepin-1-yl acid salt ) -N-isopropyl-N-phenylacetamide camforsulfonic. They were stirred (+/-) -2- (3-Amino-2,4-dioxo-5-phenyl-2,3,4,5-tetrahydrobenzo- [b] [1 , 4] diazepin-1-yl) -N-isopropyl-N-phenylacetamide (10 g) and R-camforsulfonic acid (4.98 g), in tetrahydrofuran (35 ml) and toluene (65 ml) to give a X - _ ¿___dí_ solution. The solution was heated to 70 ° C with the formation of a suspension. Water (0.4 ml) was added followed by a solution of 2-pyridinecarboxaldehyde (0.24 g) in toluene (5 ml). The mixture was heated at 70 ° C for 3 hours and then cooled to 25 ° C for 5 hours and stirred at 25 ° C for 16 hours. The suspension was suddenly cooled to a temperature of 0 to 5 ° C for a time of 1.5 hours. The solid was collected by filtration washing with toluene / tetrahydrofuran (2: 1) (10 ml). Drying in vacuo at 50 ° C afforded the title compound as a white solid (12.6 g). Chromatographic analysis: Eluent: 30% isopropyl alcohol, 70% heptane + 0.05% diethylamine; Column: 25 cm x 4.6 mm internal diameter, Chiralpak AD; Flow: 1 ml / minute; Temperature: 40 degrees centigrade; Detection: UV 230 nm; Injection Volume: 10 μL; Sample solution: 0.1 mg / ml; sample in 30% isopropyl alcohol, 70% heptane. The sample solutions were injected immediately after the preparation. Retention times: enantiomer (+), 8.2 minutes. The unwanted (-) enantiomer (12.7 minutes) was below the detection limits.

Intermediate Compound 2 3-Nitrobenzoic acid t-butyl ester Jti.

Potassium t-butoxide (3.82 g) was added to a solution of 3-nitrobenzoyl chloride (5.00 g) in anhydrous tetrahydrofuran (70 ml) and stirred under nitrogen for 2 hours. The reaction mixture was concentrated in vacuo and partitioned between dichloromethane and water. After separating the phases, the aqueous layer was back extracted with ethyl acetate. The organic layers were combined, dried over anhydrous magnesium sulfate, filtered and then concentrated in vacuo. The crude product was purified on flash silica gel, using a gradient of 0 to 5% ethyl acetate in n-hexane. The fractions containing the product were combined, concentrated in vacuo, and then dried under high vacuum to give the title compound as an oil (3.82 g). NMR X (300 MHz, CDCl 3) d = 1.63 (s, 9H); 7.62 (t, J = 7.9 Hz, 1H); 8.29-8.41 (m, 2H); 8.78-8.80 (m, 1H). MS (Cl): [M + H] + = 224.

Intermediate 3 3-Amino-benzoic acid t-butyl ester A solution of 3-nitro-benzoic acid t-butyl ester (3.77 g) in absolute ethanol (50 ml) was combined with palladium on charcoal (10% by weight). weight, 0.30 g) and stirred under atmospheric hydrogen for about 3 hours. The reaction mixture is ^^^ filtered through a pad of diatomaceous earth and then concentrated in vacuo to an oil that crystallized when dried under high vacuum to provide the title compound as a tan solid (3.28 g). X-NMR (30 MHz, CDC13) d = 1.58 (s, 9H); 6.79-6.87 (m, 1H), 7.19 (t, J = 8.5 Hz, 1H); 7.24-7.34 (m, 1H); 7.38 (d, J = 8.0 Hz, 1H). MS (Cl): [M + H] + = 194.

Intermediate 4 T-butyl ester of 3-isocyanobenzoic acid Triphosgene (13,428 g) was added to a solution of the 3-aminobenzoic acid t-butyl ester (26.50 g) and tetylamine (38.23 ml) in anhydrous tetrahydrofuran (600 ml) at a temperature of 0 to 5 ° C. The reaction mixture was stirred at a temperature of 0 to 5 ° C for 2 hours, then concentrated in vacuo to a white solid. The crude product was incorporated as a liquid slurry in hexane (500 ml), filtered, and the filtrate was concentrated in vacuo to yield the title compound as an oil (21.54 g, 71.6%). The crude isocyanate was used without further purification. X-NMR (300 MHz, CDC13) d = 1.59 (s, 9H); 7.23 (broad signal, J = 7.8 Hz, 1H); 7.36 (t, J = 7.8 Hz, 1H); 7.6 (broad signal, 1H); 7.81 (d, J = 7.8 Hz, 1H).

Intermediate 5 Ester t-butyl of (+) 3- acid. { 3- [1- (isopropyl-phenyl-carbamoylmethyl) -2,4-dioxo-5-phenyl-2,3,4,5-tetrahydro-lH-benzo [b] [1,4] diazepin-3-yl] -ureido} Benzoic Method A The intermediate (I, 66.30 g) was added slowly to a solution of 3-isocyanobenzoic acid t-butyl ester (21.54 g) in anhydrous tetrahydrofuran (750 ml). To the reaction mixture was added triethylamine (13.70 ml) dropwise. The resulting reaction mixture was stirred at room temperature overnight. The reaction mixture was poured into water (3000 ml) to yield a white solid. The solid was collected by filtration by washing with water (3 X 500 ml). Drying by vacuum filtration afforded the title compound as a white solid (65.01 g). The crude title compound was used without further purification. Chromatographic characterization: Eluent: 30% isopropyl alcohol, 70% Heptane + Diethylamine 0.05%. Column 25 cm x 4.6 mm internal diameter, Chiralpak AD; Flow rate: 1 ml / minute; Temperature: 40 degree C; Detection: UV 230 nm; injection volume: 10 μL; Sample solution: 0.1 mg / ml sample in 30% isopropyl alcohol, 70% Heptane. The sample solutions were injected immediately after preparation. Retention times: ^^^^^^^^^ ^ gk ^ ^^^^^ l ^ m ^^^^ ^^^^^^^^^^ and g ^ enantiomer (+): 15.6 minutes. The unwanted (-) enantiomer: (13.3 minutes) was below the detection limits.

Method BA a suspension of carbonyldiimidazole (13.2 g) in dichloromethane (55 ml) with stirring at 20 ° C, a solution of 3-amino-benzoic acid t-butyl ester (15.8 g) in dichloromethane was added dropwise. (40 ml) for 30 minutes. The resulting solution was stirred at 20 ° C for 1 hour. To this solution was added a solution of Intermediate Compound 1 (50 g) in dichloromethane (130 ml) for 5 minutes. The reaction was quenched by the addition of water (200 ml) and stirred for 10 minutes. The phases were separated and the organic phase was washed with water. The organic phase was concentrated at atmospheric pressure and 100 ml of dichloromethane was removed by distillation. T-butylmethyl ether (700 ml) was added and the mixture was stirred overnight at 20 ° C. The solid was collected by filtration and washed with t-butylmethyl ether (100 ml) and dried in vacuo at 45 ° C to give the title compound as a white solid (42 g, 63 mmol).

Intermediate 6 2- [(benzyloxy) imino] diethyl malonate ^^ a Di-ethyl ketomalonate (60 g) was added to 20 ° C, to a stirred suspension of O-benzylhydroxylamine (57.8 g) in IMS (500 ml) containing pyridine (30 ml). The reaction was heated to 75 ° C for 4 hours. The reaction 5 cooled and the solvents were removed under reduced pressure. The residue was partitioned between EtOAc (500 ml) and water (300 ml) and the organic layer was separated, washed with water (250 ml) and dried over MgSO4. The solvents were evaporated to give the title compound 95.3 g, as an oil 10 colorless (99%, approximately 3% w / w residual EtOAc) that was used without further purification. NMR X (300 MHz, CDC13) 7.4 (m, 5H), 5.35 (s, 2H), 4.35 (m, 4H), 1.3 (m, 6H).

Intermediate Compound 7 2- [(Benzyloxy) imino] malonic acid To a solution of intermediate 6 (40 g) in MeOH (80 ml) was added 2M NaOH (200 ml) for 20 minutes. The reaction was stirred at room temperature for 2 hours. He MeOH was removed under reduced pressure and the residual solution acidified to pH 2 by the dropwise addition of concentrated HCl (approximately 30 ml) while cooling to maintain the temperature below 35 ° C. A thick, white, liquid paste formed, which was diluted 25 with water (50 ml) to help mobility. The solids are ^^ a ^^^ collected by filtration, washed with cold water (25 ml) and dried in vacuo at 55 ° C to give the title compound as a white solid (17 g) containing about 10% w / w of residual inorganic salts. Corrected performance of approximately 45%. It was used without further purification. NMR X (300 MHz, D20) 7.4 (m, 5H), 5.2 (s, 2H).

Intermediate 8 2- [-3- [(benzyloxy) imino] -2,4-dioxo-5-phenyl-2, 3,4,5-tetrahydro-lH-l, 5-benzodiazepinyl) -N-isopropyl-N phenylacetamide Oxalyl chloride (38.3 g) was added, dropwise (for about 1 hour) to a stirred suspension of intermediate 7 (40 g, corrected for the salt content at 31.4 g) in EtOAc (200 ml) which contained DMF (0.5 ml, 5% mol). The mixture was stirred at 25 ° C for 0.5 hours and then filtered through a pad of Dicalite, washed with EtOAc (40 ml) to give a light yellow solution. The solution was added (approximately 5 minutes) to a stirred liquid paste of N-isopropyl-N-phenyl-2- (2-phenylaminophenylamino) -acetamide (50 g) in EtOAc (120 ml) at 25 ° C. The mixture heated up 60 ° C and a dark purple solution formed. After 1 hour the EtOAc (200 ml) was removed by atmospheric distillation. IPA- (120 ml) and water were added (40 ml) and the mixture was further distilled to remove more solvent (80 ml). IPA (40 ml) and water were added (40 ml) and an additional amount of solvent was distilled (80 ml). The reaction mixture was cooled to 25 ° C for 1.5 hours and the solids were collected by filtration. The solids were washed with IPA (2 x 120 ml), water (1 x 120 ml) and finally IPA again (1 x 40 ml) and then dried in vacuo at 55 ° C to give the title compound as a powder. salmon pink color (56.6 g). NMR X (300 MHz, CDC13) 2: 1 mixture of isomers around the oxime 7.6-6.95 (m, 18H), 6.9 (t, 1H), 5.3 (m, 2H), 4.95 (m, H), 4.65 ( d, 0.33H), 4.4 (d, 0.67H), 4.1 (d, 0.67H), 4.0 (d, 0.33H), 0.95 (m, 6H).

Intermediate 9 (+) -2- (3-amino-2,4-dioxo-5-phenyl-2, 3,4, 5-tetrahydrobenzo- [b] [4] diazepen-1-yl) -N-isopropyl -N-phenylacetamide To a stirred suspension of intermediate 8 (3 g) and ammonium formate (2.08 g) in IMS (30 ml) and water (3 ml) was added Pd at 5% / C (50% w / w in water) (0.25 g).

The mixture was heated under a nitrogen atmosphere at 60 ° C overnight. The hot reaction mixture is filtered through Dicalite to remove the catalyst. The catalyst was washed with hot IMS (60 ml) and filtered.

The filtrates were concentrated under reduced pressure to give the title compound as a white solid (2.34 g).

Example 1 Chromatographic Separation of Acid (+) and (-) -3- Enantiomers. { 3- [1- (isopropyl-phenyl-carbamoylmethyl) -2,4-dioxo-5-phenyl-2, 3,4,5-tetrahydro-lH-benzo [b] [1,4 | diazepin-3-yl] -ureido} benzoic. Acid 3- was prepared. { 3- [1- (isopropyl-phenyl-carbamoylmethyl) -2,4-dioxo-5-phenyl-2,3,4,5-tetrahydro-lH-benzo [b] [1,4] diazepin-3-yl] -ureido} racemic benzoic acid as described for Example 7 in US Pat. No. 5,646,140 and separated by chiral HPLC under the following conditions: column 250 x 4.0 μm (internal diameter), Diacel Chiracel OD-R 5 um; the eluent was 80: 20: 0.1: 1, 80 parts of acetonitrile, 20 parts of water, 0.1 part of triethylamine, and 1 part of acetic acid; the UV detection wavelength is 230 nm; the temperature was the environment; the flow rate was 1 ml / minute; and the injection volume was 20 ul. Under these conditions the (+) enantiomer had a retention time of 6.50 minutes 5 and the (-) isomer had a retention time of 3.89 minutes. s. ^ igjl ^ hi ^ Example 2 Acid (+) -3-. { 3- [l- (isopropyl-phenyl-carbamoylmethyl) -2,4-dioxo-5-phenyl-2, 3,4,5-tetrahydro-lH-benzo [b] [1,4] diazepin-3-yl] -ureido} benzoic Method A T-Butyl ester of (+) - 3 acid was stirred. { 3- [l- (isopropyl-phenyl-carbamoylmethyl) -2,4-dioxo-5-phenyl-2, 3,4,5-tetrahydro-lH-benzo [b] [1,4] diazepin-3-yl] -ureido} benzoic acid (intermediate 5: 65.01 g) in 4N hydrochloric acid in dioxane (280 ml) at room temperature for 6 hours. The reaction mixture was concentrated in vacuo and the resulting solid / oil was triturated in water to yield a white solid. The white solid was collected by filtration and washed with water (2 X 500 ml). The crude product was dissolved in hot acetone (250 ml) and water (275 ml) was added until the solution became turbid. Additional acetone (40 ml) was added and the solution was heated until a clear solution was obtained. The solution was left aside and allowed to cool. The resulting white solid was collected by filtration, washed with water (3 X 100 ml) and dried under a vacuum hood (50.8-63.5 cm Hg) at a temperature of 40 to 50 ° C, to provide the title compound as a white solid (40,496 g). Its purity was analyzed by chiral chromatography (see the chromatographic separation protocol). ^ -'- - * • * - »- •" - ~ &- -? I ?? í ?? jj¡M¿lt Optical rotation (0.721 g in 100 mL of acetone) [a] D = +84.3. NMR AH (300 MHz, DMSO) d = 0.95 (d, J = 7.5 Hz, 3H), 0.97 (d, J = 7.2 Hz, 3H), 4.18 (d, J = 16.7 Hz, '1H), 4.48 (d) , J = 16.7 Hz, 1H), 4.78 (m, 1H), 5.02 (d, J = 7.8 Hz, 1H), 6.91 (d, J = 7.8 Hz, 1H), 6.95 (broad signal, J = 8.1 Hz, 1H), 7.22-7.57 (m, 19H), 8.00 (s, 1H), 9.34 (s, 1H), 12.78 (s, 1H), MS (ES): [M + l] = 606.1; [M + Na] ] = 628.1; [Ml] = 604.1.

Method B Intermediate 5 (5 g) was added to a mixture of acetone (15 ml) and formic acid (25 ml) at room temperature and the resulting suspension was heated to 55 ° C and stirred for 5 hours. Water (30 ml) was added, drop by drop, to the solution, to ensure that the temperature of the content was maintained above 50 ° C. The resulting liquid paste was stirred at 55 ° C for 1 hour, cooled to room temperature and stirred overnight. The liquid paste was filtered and washed with water (3 x 25 ml). The residue was added to IMS (50 ml) and the liquid paste was heated to 45 ° C and stirred overnight. The liquid paste was cooled to room temperature, filtered and the wet cake dried in vacuo at 55 ° C to give the title compound as a white solid (3.40 g). ^^ & Chiral chromatographic analysis showed that the required product of the reaction contained 0.7% of the undesired (-) enantiomer. 5 Examples of Pharmacy Oral Solution Active Ingredient 0.5-800 mg Polyethylene glycol 400 NF c.s.p. 50 ml The active ingredient is suspended in Polyethylene Glycol 400 and then dissolved by application of sonic energy, to produce the oral solution.

Oral Suspension Active Ingredient 0.5-80 mg 15 Polysorbate 80 NF (Tween 80) 0.02 ml Sterile Water for Irrigation c.s.p. 20 ml The active ingredient is added to a solution of Tween 80 (20 ml) at 0.1% (v / v) and then sonic energy is applied to the mixture to produce the oral suspension. 20 Tablets a. Active ingredient 6 mg Lactose anhydrous USP 136.2 mg Sodium glycolate starch USP / NF 6 mg 25 Stearic acid USP / NF 1.5 mg ss -am -U. - Colloidal silicon dioxide USP / NF _ 0.3 mg. Compression Weight 150 mg The active ingredient, lactose, and sodium starch glycolate are screened through a 590 micron sieve and mixed in a suitable mixer. Stearic acid (sieved through a 250 micron sieve) and colloidal silicon dioxide are added and mixed with the active mixture. The mixture is compressed to form tablets using suitable punches. b. Active ingredient 6 mg Microcrystalline cellulose USP / NF 136.5 mg Crospovidone USP / NF 6 mg Magnesium stearate USP / NF 1.5 mg Compression weight 150 mg The active ingredient, microcrystalline cellulose and crospovidone are sieved through a 590 micron sieve and mixed in a suitable mixer. The magnesium stearate is sieved (through a 250 micron sieve) and mixed with the active mixture. The resulting mixture is compressed to form tablets using punches suitable for tablets.

Capsules a. Active ingredient 6 mg llii 11 ^ Microcrystalline cellulose USP / NF 128.25 mg Sodium glycolate starch USP / NF 15 mg Magnesium stearate USP 0.75 mg Filling weight 150 mg The active ingredient, microcrystalline cellulose, and sodium starch glycolate, sieve through a sieve with 590 micron mesh, mixed together and lubricated with magnesium stearate that had been sieved through a 250 micron sieve. The mixture is filled into capsules of a suitable size. b. Active ingredient 6 mg Lactose monohydrate USP 130.5 mg Povidone USP 6 mg Crospovidone NF 6 mg Magnesium stearate 1.5 mg Filling weight 150 mg The active ingredient and the lactose are mixed together and granulated with a solution of Povidone. The wet mass is dried and ground. Magnesium stearate and Crospovidone are screened through a 250 micron sieve and mixed with the granules. The resulting mixture is filled into hard gelatin capsules of a suitable size. ^^^^? a ?? t ^? ^ íá? ^^ áß? l ^ t ?? .-ni i-i't. - BIOLOGICAL TESTS The (+) and (-) enantiomers and the racemic mixture were characterized in the following tests. The results of these tests are summarized in the following table.

Preparation of Gallbladder Gallbladder Tissue Male Hartley guinea pig bile vesicles, sacrificed with C0 = atmosphere, were removed. The adherent connective tissue was removed from the biliary vesicles and cut into two rings of each animal (2 to 4 mm in length). The rings were suspended in organ chambers containing a physiological saline solution (118.4 mM NaCl, 4.7 mM KCl, 1.2 mM MgSO4, 2.5 mM CaCl2, 1.2 mM KH2P03, 25 mM NaHCO3, 11.1 mM dextrose). The bath solution was maintained at 37 ° C and was aerated with 95% 02/5% C02 to maintain a pH = 7.4. The fabrics were connected by gold chains and mounting wires, stainless steel, to isometric force displacement transducers (Grass, Model FT03 D). The answers were recorded in a polygraph (Grass, Model 7E). A tissue from each animal served as a time / solvent control and did not receive the test compound. The rings were gradually stretched (for a period of 120 minutes) to a resting basal tension of 1 gm which was maintained throughout the experiment. During the period of adjustment of the basal tension, the rings were exposed - to -acetylcholine (10 ~ D M) four times, to verify tissue contractibility. The tissues were then exposed to a dose below the maximum dose of sulfated CCK-8 (Sigma, 3 X 10 ~ 9 M). After obtaining a stable response, the tissues were washed 3 times rapidly and after 5 to 10 minutes, for 1 hour, to reestablish a stable baseline. 10 EC5o Values of the Agonist. The compounds were dissolved in dimethylsulfoxide (DMSO) then diluted with water and assayed through a cumulative concentration-response curve for the test compound (10"11 to 3 X 10 ~ 5 M) followed by a 15 concentration-response curve for sulfated CCK-8 (10 ~ 10 to 10"° M) in the presence of the highest concentration of the test compound.As a final test, acetylcholine (1 mM) was added to induce the maximum contraction For each test compound a minimum of 20 three activity determinations.

Evaluation of cell lines containing the stable CCK receptor. Clones of cDNA for receptors CCK-A18 or 25 human CCK-B19, were ligated into the pcDNA-Neo vector of ^^ nm __ ^^?, U ...; ^ Invitrogen Corp (San Diego, CA) - for direct transfection. The DNA was prepared by the alkaline lysis method and transfected into 'CHO-Kl cells (ATCC, Rockville, MD) using the Lipofectin24 reagent (Gibco BRL, Gaithersberg, MD). Stable transfectants were initially selected by the use of Geneticin (Gibco BRL) and the resistant cells containing the receptor were enriched by fluorescence-activated cell sorting, based on the binding of Fluorescein-Gly- [(Nle28, 31) -CCK -8 The clonal lines were subsequently established by the limiting dilution method.

Preparation of cell membranes. CHO-Kl cells stably transfected with cDNA from the CCK-A or CCK-B receptor were cultured at 37 ° C under a humidified atmosphere (95% 02/5% C02) in Ham's F12 medium supplemented with 5% serum Fetal bovine thermally inactivated. Cells were subcultured twice weekly and cultured to a density of 2 to 4 million cells / mL. Cells were harvested by centrifugation (600 X g, 15 minutes, 4 ° C) and resuspended in buffer (20 mL, pH 7.4) containing TrisHCl (25 mM), EDTA (5 mM), EGTA (5 mM), phenylsulfonyl fluoride (0.1 mM) and soybean trypsin inhibitor (100 μg / mL). The cells were broken with a homogenizer Teflon and motorized glass were added (25 times) and the homogenate was centrifuged at low speed (600 X g, 10 minutes, 4 ° C). The supernatant was collected and centrifuged at high speed (500,000 X g, 15 minutes, 4 ° C) to form pellets of the particulate fraction. The pellets at low speed were processed three additional times. The particle fractions were combined and resuspended in buffer (1-5 mg protein / mL) and frozen -80 ° C. The protein concentration was determined according to the manufacturer's instructions, using BioRad reagent and bovine serum albumin as a standard.

Receptor Binding Assays. 125I-Bolton Hunter CCK-8 was dissolved (Amersham, 2000 Ci / mmol) in buffer for binding (pH 7.4, 100,000 cpm / 25 μL) containing HEPES (20 mM), NaCl (118 mM), KCl (5 mM), MgCl2 (5 mM) and EGTA (1 mM) . The non-specific binding was determined with MK-32920 (10 μM, CCK-A) or L-365,26021 (10 μM, CCK-B). The test compounds were dissolved in DMSO at a concentration of 100 times the concentration of the final assay and diluted to appropriate concentrations with buffer for binding. Binding assays were performed in triplicate using 96-well plates, at , _, _, Which were sequentially added the following: test compound (25 μL), i25I-Bolton Hunter CCK-8 (25 μL), buffer (pH 7.4, 150 μL) and receptor preparation (50 μL) . The final concentration of DMSO was 1% in all the test wells. After 3 hours at 30 ° C the incubation was terminated by rapid filtration of the mixture over glass filters (Whatman GF / B) with subsequent washing to remove the unbound ligand. The binding radioactivity was quantified by gamma counting.

Intracellular Calcium Measurements: Stably transfected CHO-Kl cells were cultured with hCCK-A or hCCK-B receptors, on glass coverslips, to a confluence of 75-90%. The cells were loaded for 50 minutes in serum-free culture medium, containing 5 mM FURA2-AM and 2.5 mM probenecid. A calcium analyzer, JASCO CAF-102, was used to measure changes in intracellular calcium concentration, using standard radiometric techniques, using excitation wavelengths of 340 nm and 380 nm. The cells were perfused with increasing concentrations of CCK-8 (n = 2) or compounds (n = 2) until a value was obtained in the ratio of 340/380. A wash / recovery period of at least 10 minutes between successive stimulations was allowed. The maximum response was normalized to the maximum response induced by CCK-8. The EC50 were calculated at the concentration required to induce the half-maximal response. In addition to the agonist concentration-response curves, CHO-Kl cells expressing human CCK-B were perfused for 1 hour with three concentrations of the compounds (10 ~ 8, 10"7, 10 ~ 6 M, n = 2), then response curves were obtained for the concentration, for CCK-8 (from 10 ~ 12 to 10"6 M). 10 Anorexia Tests: Male Long-Evans Rats (225-300 g) were conditioned for two weeks to consume an appetizing liquid diet (Bio-Serve F1657, Frenchtown, NJ) after a 15 fasting for 2 hours. On the day of pretreatment the rats were fasted (100 minutes) and injected with the drug vehicle (propylene glycol, PG, 1 mL / kg) and oral saline preload (0.9% NaCl, 8 mL / kg). ). Access to the liquid diet was allowed for 20 minutes 20 after and the consumption was measured at 30, 90 and 180 minutes. To qualify for the treatment study with the drug, the rats had to consume at least 8 mL of liquid diet within the first 30 minutes on the day of pretreatment. The next day, followed by the deprivation of 100 minutes, 25 rats (from 8 to 10 animals per dose) were treated IP or PO ? rf ^^ ... ^ ........ ¿Jai ».- *, > . with the vehicle (PG, 1 mL / kg) or several doses (from 0.01 to 10 μmol / kg) of the test compound dissolved in PG (1 mL / kg), immediately followed by the oral preload of saline. The access to the food was allowed again, 20 minutes later, and the food consumption was measured at 30, 90 and 180 minutes. All the feed consumption data were normalized for each rat, at the respective values of the pre-treatment day. Power was determined at 30 minutes and efficacy at 30 minutes, dose of 1 μmol / kg.

Gallbladder emptying test in mouse: Makovec, F .; Bani, M .; Cereda, R .; Chiste, R .; Paccini, M. A .; Revel, L .; Rovati, L. C. Antispasmodic Activity on the Gallbladder of the Mouse of CR1409 (Lorglumide), a Potent Antagonist of Peripheral Cholecystokinin. Pharmacol. Res. Commun. 1987, 19, 41-51.

PApp Values: A measurement of intestinal permeation determined using the in vi tro assay of Artursson P. and Karlson J. 1991, Biochem. Biophys Res. Common. 175, 880-885 (Correlation between the absorption of oral drugs in humans and the apparent permeability coefficients of the drug, in epithelial, intestinal, human cells (CACO-2)).

Pharmacological Comparison of Enantiomers and Racemates 10 fifteen twenty 1. Selectivity Ratio = IC50 (hCCK-A) / Kx (hCCK-B) 2. Inactive up to 10 μmol / kg 25 3. There was no significant reduction in feeding, observed with doses up to 10 μmol / kg. ^ tt ^ g ^ 1 ^ '- ~ "^ -lililí * *" * Three unexpected biological activities distinguish the (+) enantiomer of the (-) enantiomer and the racemate. Two of these activities refer to the improved efficacy of CCK-A, which would have improved the beneficial activity of this enantiomer. The third one refers to the antagonist activity towards CCK-B, of the (+) enantiomer, which would have proven beneficial due to its lower toxicity. The (+) enantiomer was four times more potent than the racemate in the isolated guinea pig gallbladder test, in vi tro, ("GPGB"). The (+) enantiomer was eight times more potent than the racemate in the mouse gall bladder emptying assay (oral dosing). It is expected that this increased potency will be beneficial in the treatment of gallbladder stasis and in the treatment of obesity, since stasis of the gallbladder is a critical problem with rapid weight loss. Anorectic agents are used chronically and therefore it is essential that they possess minimal risk of toxicity. The primary toxicity associated with the use of cholecystokinin is concomitant with the agonist activity of the CCK-B receptor. Activation of the CCK-B receptor is mainly associated with increased anxiety and increased gastric acid secretion. The utility of antagonists of CCK-B has been explored, both for the U ^ ifa_É__ÉÉUÍiii development of anxiolytic agents as antiulcer agents. See, for example, Lo e, J. "Cholecystokinin- B Receptor Antagonists" in Exp. Opin. Ther. Patents, 5 (3), pages 231-237 (1995). 5 The subtype of the CCK receptor, predominant in the rodent pancreas, is the CCK-A subtype, and the activation of this subtype induces pancreatic hyperstimulation and hypertrophy in rodents. Both of these activities are considered undesirable. Recently it has been reported 10 tissue distribution of CCK receptors in human tissues. Surprisingly, the predominant receptor subtype in the human pancreas is the subtype of the CCK-B receptor. See, for example, Wank, S.A., "Cholecystokinin Receptors" in American Journal of 15 Physiology - Gastrointestinal & Liver Physiology, 32 (5) :, pages G628-G646, (1995). Thus, in humans, the activation of the CCK-B receptor (agonist activity CCK-B) could induce increased gastric acid anxiety and secretion, as well as hyperstimulation and pancreatic hypertrophy with 20 the prolonged use. To decrease the risk of agonist activity of CCK-B, undesirable, in vivo, the preferred compound should have affinity for the CCK-B receptor and have an antagonistic activity by CCK-B, measurable, in in vitro assays. 25 vi tro. Both enantiomers and the racemate are antagonists of the CCK-B. Although all three compositions have affinities (IC5o) and similar efficiencies (EC50) for the CCK-A receptor, the (+) enantiomer has the affinity (IC5u) and selectivity (46 times) higher for the CCK-B receptor. Thus, the (+) enantiomer is preferred both in terms of potency and efficacy of CCK-A, as well as in terms of the minimum potency with respect to the toxic side effects induced by CCK-B. The (+) enantiomer of the invention is essentially non-toxic at the therapeutically useful doses. For example, in oral studies with a single dose, it was found that the maximum non-lethal dose was higher than 2000 mg / kg in rats and 1000 mg / kg for male mice and 500 mg / kg for female mice.

It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is the conventional one for the manufacture of the objects or products to which it refers.

Claims (12)

1. CLAIMS Having described the invention as above, the content of the following claims is claimed as property: 1. The compound characterized in that it is 3- acid. { 3- [1- (isopropyl-phenyl-carbamoylmethyl) -2,4-dioxo-5-phenyl-2,3,4,5-tetrahydro-lH-benzo [b] [1,4] diazepin-3-yl] ureido} enantiomerically enriched benzoic acid, or a pharmaceutically acceptable salt or solvate thereof. 2. The enantiomerically enriched compound according to claim 1, characterized in that the (+) enantiomer, or a pharmaceutically acceptable salt or solvate thereof, is at least 90% of the compound. 3. The enantiomerically enriched compound according to claim 1 or claim 2, characterized in that the (+) enantiomer, or a pharmaceutically acceptable salt or solvate thereof, is at least 99% of the compound. 4. A pharmaceutical composition characterized in that it comprises the enantiomerically enriched compound according to any of claims 1 to 3, in a mixture with one or more pharmaceutically acceptable carriers and / or excipients. A method for the treatment of a disease or condition mediated by CCK-A, characterized in that it comprises the administration of an effective amount of a compound in accordance with any of the 5 claims 1 to 3. 6. A method for the treatment of a disease or condition mediated by CCK-A, characterized in that it comprises the administration of the pharmaceutical composition according to claim 4. 10 7. The method according to the claim 5 or with claim 6, characterized in that the disease or condition is obesity, stasis of the gall bladder, or diabetes. 8. The method according to claim 5 or claim 6, characterized in that the disease or condition is obesity. 9. The use of a compound according to any of claims 1 to 3 in the manufacture of a medicament for the treatment of a disease or 20 condition mediated by CCK-A. A process for the preparation of a compound according to claim 1, characterized in that it comprises: (a) the separation of 3- [3- [1- (isopropyl-phenyl-25-carbamoylmethyl) -2,4-dioxo acid -5-phenyl-2, 3,4,5- racemic tetrahydro-lH-benzo [b] [1,4] diazepin-3-yl] benzoic, by chiral high-performance liquid chromatography; (b) the reaction of the appropriate enantiomer of the amine of formula (II). With the isocyanate of formula (III), imidazolide of formula (IV) or optionally substituted phenyl carbamate of formula (V), followed by removal of the protective group 20 carboxi, R. MMa &riaaMaM? I laMÉ ^^^^^^^^^^ (^^ ¿¡^^^^^^^^^^^^^ (V) 11. A process according to claim 10, characterized in that the required compound of claim 1 is prepared by the racemic amine (II) which has been prepared by the concomitant reduction and hydrogenolysis of the oxime (VI), (SAW) wherein R2 is an optionally substituted benzyl group, ^^ - * - * - - - - - 12. A process according to claim 11, characterized in that the oxime (VI) is prepared from the orthophenylenediamine (VII) and an activated derivative of the diacid (VIII), Wherein R2 is an optionally substituted benzyl group,