EP1212305A2 - Derives de 1,5-benzodiazepines - Google Patents

Derives de 1,5-benzodiazepines

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Publication number
EP1212305A2
EP1212305A2 EP00929488A EP00929488A EP1212305A2 EP 1212305 A2 EP1212305 A2 EP 1212305A2 EP 00929488 A EP00929488 A EP 00929488A EP 00929488 A EP00929488 A EP 00929488A EP 1212305 A2 EP1212305 A2 EP 1212305A2
Authority
EP
European Patent Office
Prior art keywords
cck
compound
enantiomer
phenyl
formula
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP00929488A
Other languages
German (de)
English (en)
Inventor
David Colclough
Anne Hodgson
Jerzy Ryszard Szewczyk
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Glaxo Group Ltd
Original Assignee
Glaxo Group Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from GBGB9910366.5A external-priority patent/GB9910366D0/en
Priority claimed from GBGB0008179.4A external-priority patent/GB0008179D0/en
Application filed by Glaxo Group Ltd filed Critical Glaxo Group Ltd
Publication of EP1212305A2 publication Critical patent/EP1212305A2/fr
Withdrawn legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D243/00—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms
    • C07D243/06—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms having the nitrogen atoms in positions 1 and 4
    • C07D243/10—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms having the nitrogen atoms in positions 1 and 4 condensed with carbocyclic rings or ring systems
    • C07D243/12—1,5-Benzodiazepines; Hydrogenated 1,5-benzodiazepines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/55—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole
    • A61K31/551—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole having two nitrogen atoms, e.g. dilazep
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/04—Anorexiants; Antiobesity agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Definitions

  • This invention relates to novel 1 ,5-benzodiazepine derivatives, to processes for their preparation, to pharmaceutical compositions containing them and to their use in medicine. More particularly, it relates to compounds which exhibit agonist activity for CCK-A receptors.
  • Cholecystokinin is a peptide found in the gastrointestinal tract and 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, N.Y., 169.
  • CCK has been implicated inter alia as a physiological satiety hormone involved in appetite regulation, see Della-Ferra et al, Science, 206, 471 (1979), Saito et al., Nature, 289, 599, (1981), G.P.
  • CCK-A and CCK-B Two subtypes of CCK receptors have been identified, designated as CCK-A and CCK-B, and both have been found in the periphery and central nervous systems. It has recently been reported that CCK-B receptors are similar to the gastrin receptor, see Pisegna, J.R., de Weerth, A, Huppi, K, Wank, S.A., Biochem. Biophys. Res. Commun. 189, 296-303 (1992). CCK-A receptors are located predominantly in peripheral tissues including the pancreas, gallbladder, ileum, pyloric sphincter and vagal afferent nerve fibers; CCK-A receptors are found to a lesser extent in the brain, see T.H.
  • CCK agonist activity has been linked to inhibition of food intake in animals and thus weight loss, see Della-Fera, et al, supra, K.E. Asin, et al, Intl. Conference on Obesity, abstract pp.40 (1990). It has been suggested that CCK acts in the periphery through vagal fibers and not directly on the brain to produce satiety, see Smith, G.P. and Cushin, B.J., Neuroscience Abstr., 4, 180 (1978), Smith, G.P., Jerome, C, Cushin, B.J., Eterno, R., and Simansky, K.J., Science, 212, 687-689, (1981 ).
  • U.S. Patent No. 5,646,140 discloses certain 3-amino 1 ,5- benzodiazepine compounds which exhibit agonist activity for the CCK-A receptor thereby enabling them to modulate the hormones gastrin and cholecystokinin (CCK) in mammals. See in particular, the compound of Example 7. Certain of these compounds also exhibit antagonist activity at CCK-B receptors.
  • the present invention provides an enantiomerically enriched compound of Formula (I) or a pharmaceutically acceptable salt or solvate thereof.
  • the compound of Formula (I) is 3- ⁇ 3-[1-(lsopropyl-phenyl-carbamoylmethyl)-2,4- dioxo-5-phenyl-2,3,4,5-tetrahydro-1 H-benzo[b][1 ,4]diazepine-3-yl]-ureido ⁇ benzoic acid.
  • This compound has a chiral carbon on the benzodiazepine ring.
  • Applicants have found that the enantiomer which rotates light in the positive direction, under the conditions described below, is preferred.
  • This enantiomer which is hereinafter referred to as the (+) enantiomer has the (S) configuration according to the Cahn Ingold Prelog convention.
  • enantiomerically enriched means that there is more of the (+) enantiomer than the (-) enantiomer as opposed to the racemic mixture which has equal amounts of each isomer.
  • the compound of this invention or “the enantiomerically enriched compound of this invention”, and expressions containing these or similar phrases, include pharmaceutically acceptable salts and solvates thereof.
  • the “(+) enantiomer” refers to the optical rotation of the enantiomer and not to salts and solvates thereof. 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.
  • 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 exhibits CCK-A agonist activity and can be considered a full cholecystokinin agonist in that it binds to CCK-A receptors and fully stimulates gallbladder contraction and reduces feeding in animal paradigms.
  • (+) enantiomer of this invention should be useful for the treatment of obesity as well as related pathologies, such as hypertension, gallbladder stasis, and diabetes, indirectly through weight loss and directly through CCK-A mediated delayed gastric emptying.
  • the (+) enantiomer disclosed herein provides for new approaches for inducing satiety, providing for appetite regulation and modifying food intake in mammals, especially humans, to regulate appetite, treat obesity and maintain weight loss.
  • a method for the treatment, in a mammal, including man, of a CCK-A mediated disease or condition comprising administering to the patient a therapeutically effective amount of the (+) enantiomer of this invention.
  • the present invention provides the use of the enantiomerically enriched compound of this invention or a pharmaceutically acceptable salt or solvate thereof for the manufacture of a medicament for the treatment of CCK-A medicated diseases or conditions.
  • treatment extends to prophylaxis as well as the treatment of established diseases or symptoms.
  • amount of the preferred enantiomer of the invention required for use in treatment will vary with the nature of the condition being treated and the age and the condition of the patient and will be ultimately at the discretion of the attendant physician or veterinarian.
  • doses employed for adult human treatment will typically be in the range of 0.02 - 5000 mg per day, e.g., 1-1500 mg per day.
  • the desired dose may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, for example as two, three, four or more sub-doses per day.
  • the enantiomerically enriched compound of the present invention may be therapeutically administered as the raw chemical, it is preferable to present the active ingredient as a pharmaceutical composition. Accordingly, the present invention further provides for a pharmaceutical composition comprising the enantiomerically enriched compound of this invention together with one or more pharmaceutically acceptable carriers and/or excipients therefore and, optionally, other therapeutic and/or prophylactic ingredients.
  • the carrier(s) and/or excipients therefor must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.
  • Formulations of the present invention include those especially formulated for oral, buccal, parenteral, implant, topical or rectal administration, however, oral administration is preferred.
  • buccal administration the composition may take the form of tablets or lozenges formulated in conventional manner.
  • Tablets and capsules for oral administration may contain conventional excipients such as binding agents, (for example, syrup, acacia, gelatin, sorbitol, tragacanth, mucilage of starch or polyvinylpyrrolidone), fillers (for example, lactose, sugar, microcrystalline cellulose, maize-starch, calcium phosphate or sorbitol), lubricants (for example, magnesium stearate, stearic acid, talc, polyethylene glycol or silica), disintegrants (for example, potato starch or sodium starch glycollate) or wetting agents, such as sodium lauryl sulphate.
  • the tablets may be coated according to methods well-known in the art, including enteric coatings.
  • the preferred enantiomer of the present invention may be incorporated into oral liquid preparations such as aqueous or oily suspensions, solutions, emulsions, syrups or elixirs, for example.
  • formulations containing these the preferred enantiomer may be presented as a dry product for constitution with water or other suitable vehicle before use.
  • Such liquid preparations may contain conventional additives such as suspending agents such as sorbitol syrup, methyl cellulose, glucose/sugar syrup, gelatin, hydroxyethylcellulose, carboxymethyl cellulose, 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 sorbic acid.
  • Such preparations may also be formulated as suppositories, e.g., containing conventional suppository bases such as cocoa butter or other glycerides.
  • compositions the present invention may be formulated for parenteral administration by injection or continuous infusion.
  • Formulations for injection may take such forms as suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents.
  • the active ingredient may be in powder form for constitution with a suitable vehicle (e.g., sterile, pyrogen-free water) before use.
  • composition according to the invention may also be formulated as a depot preparation.
  • Such long acting formulations may be administered by implantation (for example, subcutaneously or intramuscularly) or by intramuscular injection.
  • the preferred enantiomer of the invention may be formulated with suitable polymeric or hydrophobic materials (as an emulsion in an acceptable oil, for example), ion exchange resins or as sparingly soluble derivatives as a sparingly soluble salt, for example.
  • compositions according to the invention may contain between 0.1 - 99% of the active ingredient, conveniently from 30 - 95% for tablets and capsules, and 3 - 50% for liquid preparations.
  • the (+) enantiomer of this invention can made by first making the racemic mixture as described in Example 7 in U.S. Patent No. 5,646,140 and then separating the enantiomers by chiral chromatography.
  • the (+) enantiomer may be prepared by reaction of the appropriate enantiomer, namely the (S)-enantiomer of the amine of formula (II);
  • the reaction conveniently takes place in the presence of a suitable solvent such as an ether (e.g. tetrahydrofuran) or a halohydrocarbon (e.g. dichloromethane) or nitrile (e.g. acetonitrile) at a temperature in the range of 0-80°C.
  • a suitable solvent such as an ether (e.g. tetrahydrofuran) or a halohydrocarbon (e.g. dichloromethane) or nitrile (e.g. acetonitrile)
  • a suitable solvent such as an ether (e.g. tetrahydrofuran) or a halohydrocarbon (e.g. dichloromethane) or nitrile (e.g. acetonitrile)
  • the required enantiomer of the amine (II) may be used in the form of a salt thereof e.g. R-camphorsulphonic acid salt and in this embodiment the
  • the hydrolysis of the carboxyl protecting group may be carried out using conventional procedures. (Protecting groups in Organic Synthesis T. Greene, Ed, Wiley Interscience, New York, p168, 1981 ). Thus for example when R is a t-butyl group this may be removed by hydrolysis with an appropriate acid such as hydrochloric acid, trifluoroacetic acid or formic acid using established procedures. For example by reaction with hydrochloric acid in a solvent such as 1 ,4-dioxane, or by reaction with formic acid in a solvent such as acetone or aqueous acetone and with heating.
  • racemic amine (II) can be prepared by the method described in Intermediate 11 of US Patent No. 5,646,140.
  • the racemic amine (II) may be prepared by concomitant reduction and hydrogenolysis of the oxime (VI) wherein R 2 is an optionally substituted benzyl group.
  • the reaction is conveniently carried out using a suitable palladium catalyst e.g. palladium on carbon e.g. 5% Pd/on charcoal in the presence of hydrogen or aqueous ammonium formate and in a solvent such as an aqueous alkanol e.g. ethanol, isopropanol or industrial methylated spirits, or tetrahydrofuran.
  • a suitable palladium catalyst e.g. palladium on carbon e.g. 5% Pd/on charcoal
  • a solvent such as an aqueous alkanol e.g. ethanol, isopropanol or industrial methylated spirits, or tetrahydrofuran.
  • heating e.g. 40-80° such as 60°C.
  • R 2 groups for use in the reaction include benzyl, or substituted benzyl, such as p-methoxybenzyl, or benzhydryl.
  • the oxime (VI) may be prepared by reaction of the ortho-phenylene diamine derivative (VII)
  • the activated derivative of the di acid (VIII) is the corresponding diacyl halide e.g. chloride and this is prepared in situ by reaction of the di acid (VIII) with an oxalyl halide e.g. oxalyl chloride.
  • the reaction is conveniently carried out on an aprotic solvent such as an ester e.g. ethyl acetate, toluene, dichloromethane, dimethoxyether or mixtures thereof and in the presence of dimethylformamide.
  • the di acid (VIII) is conveniently prepared by reaction of a di alkylketomalonate e.g. diethyl ketomalonate with the corresponding hydroxylamine R 2 ONH 2 in a solvent such as an alkanol e.g. ethanol or industrial methylated spirits and in the presence of a base e.g. pyridine, followed by hydrolysis of the corresponding di alkyl-oximino malonate using aqueous sodium hydroxide.
  • a di alkylketomalonate e.g. diethyl ketomalonate
  • R 2 ONH 2 a solvent
  • a base e.g. pyridine
  • 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).
  • Isocyanates of Formula (III) may be purchased or prepared by the reaction of the corresponding amine (VI) with phosgene or triphosgene in a suitable solvent such as methylene chloride.
  • Imidazolides of Formula (IV) can be prepared by treatment of the corresponding amine (VI) with carbonyl diimidazole in a suitable solvent (dichloromethane, ether, tetrahydrofuran) at a temperature ranging from 0
  • the optionally substituted phenyl carbamates of Formula (V) can be prepared by the 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 at a temperature of 0 - 50° C.
  • a base pyridine, triethylamine
  • a suitable solvent dichloromethane
  • EtOAc ethyl acetate
  • MeOH methanol
  • DMF N, N-dimethylformamide
  • IPA isopropyl alcohol
  • IMS industrial methylated spirits.
  • (+/-)-2-(3-Amino-2,4-dioxo-5-phenyl-2,3,4,5-tetrahydrobenzo-[b][1 ,4]diazepin-1- yl)-N-isopropyl-N-phenylacetamide (10g) and R-camphorsulfonic acid (4.98g) were stirred in tetrahydrofuran (35ml) and toluene (65ml) to give a solution. The solution was heated to 70°C with formation of a suspension. Water (0.4ml) was added followed by a solution of 2-pyridinecarboxaldehyde (0.24g) in toluene (5ml).
  • Triphosgene (13.428g) was added to a solution of 3-amino-benzoic acid t-butyl ester (26.50g) and triethylamine (38.23ml) in anhydrous tetrahydrofuran (600 ml) at 0-5 °C.
  • the reaction mixture was stirred at 0-5 °C for 2h, then concentrated in vacuo to a white solid.
  • the crude product was slurried in hexane (500 ml), filtered, and the filtrate was concentrated in vacuo to afford the title compound as an oil (21.54 g, 71.6%).
  • the crude isocyanate was used without further purification.
  • Intermediate (I; 66.30g) was slowly added to a solution of 3-lsocyano-benzoic acid t-butyl ester (21.54g) in anhydrous tetrahydrofuran (750 ml). Triethylamine (13.70ml) was added dropwise to the reaction mixture.
  • Di-ethylketomalonate 60g was added at 20°C to a stirred suspension of O- benzylhydroxylamine (57.8g) in IMS (500ml) containing pyridine (30ml). The reaction was heated at 75°C for 4hr. The reaction was cooled and solvents removed under reduced pressure. The residue was partitioned between EtOAc (500ml) and water (300ml) and the organic layer separated, washed with water (250ml) and dried over MgSO 4 . Solvents were evaporated to give the title compound 95.3g, as a colourless oil (99%th, ca 3%w/w residual EtOAc) which was used without further purification.
  • Oxalyl chloride (38.3g) was added dropwise ( ⁇ 1 hr) to a stirred suspension of Intermediate 7 (40g, corrected for salt content to 31.4g) in EtOAc (200ml) containing DMF (0.5ml, 5 mol%). The mixture was stirred at 25°C for 0.5 hour then filtered through a pad of Dicalite, washing with EtOAc (40ml) to give a clear yellow solution. The solution was added ( ⁇ 5mins) to a stirred slurry of N- isopropyl-N-phenyl-2-(2-phenylaminophenylamino)-acetamide (50g) in EtOAc (120ml) at 25°C.
  • the reaction mixture was concentrated in vacuo and the resulting solid/oil was triturated in water to afford 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 cloudy. Additional acetone (40 ml) was added and the solution was heated until a clear solution was obtained. The solution was set aside and allowed to cool.
  • the resulting white solid was collected by filtration washed with water (3 X 100 ml) and dried under house vacuum (20-25 in Hg) at 40-50 °C to provide the title compound as a white solid (40.496g).
  • Active ingredient 0.5-800 mg Polyethylene glycol 400 NF q.s. to 50 ml
  • the active ingredient is suspended in Polyethylene glycol 400 and is then dissolved by sonication to produce the oral solution.
  • the active ingredient is added to a 0.1 % (v/v) Tween 80 solution (20 ml) and the mixture is then sonicated or shaken to produce the oral suspension.
  • the active ingredient, lactose, and sodium starch glycolate are sieved through a 590 micron sieve and blended in a suitable mixer.
  • Stearic acid (screened through a 250 micron sieve) and colloidal silicon dioxide are added to and blended with the active blend.
  • the blend is compressed into tablets using suitable punches.
  • the active ingredient, microcrystallline cellulose and crospovidone are sieved through a 590 micron sieve and blended in a suitable blender.
  • the magnesium stearate is screened (through a 250 micron sieve) and blended with the active blend.
  • the resultant blend is compressed into tablets using suitable tablet punches.
  • the active ingredient, microcrystalline cellulose, and sodium starch glycolate are screened through a 590 micron mesh sieve, blended together and lubricated with magnesium stearate, that has been screened through a 250 micron sieve.
  • the blend is filled into capsules of a suitable size.
  • the active ingredient and lactose are blended together and granulated with a solution of Povidone.
  • the wet mass is dried and milled.
  • the magnesium stearate and Crospovidone are screened through a 250 micron sieve and blended with the granule.
  • the resultant blend is filled into hard gelatin capsules of a suitable size.
  • (+) and (-)-enantiomers and the racemic mixture were characterized in the following assays.
  • the results of these assays are summarized in the table below.
  • Gallbladders were removed from male Hartley guinea pigs sacrificed with CO2 atmosphere.
  • the isolated gallbladders were cleaned of adherent connective tissue and cut into two rings from each animal (2-4 mm in length).
  • the rings were suspended in organ chambers containing a physiological salt solution (118.4 mM NaCI, 4.7 mM KCI, 1.2 mM MgS04, 2.5 mM CaCl2, 1.2 mM KH2PO3, 25 mM NaHC03 , 11.1 mM dextrose).
  • Tissues were connected via gold chains and stainless steel mounting wires to isometric force displacement transducers (Grass, Model FT03 D). Responses were then recorded on a polygraph (Grass, Model 7E).
  • One tissue from each animal served as a time/solvent control and did not receive test compound. Rings were gradually stretched (over a 120-min. period) to a basal resting tension of 1 gm which was maintained throughout the experiment. During the basal tension adjustment period, the rings were exposed to acetylcholine (10 ⁇ 6 M) four times to verify tissue contractility. The tissues were then exposed to a submaximal dose of sulfated CCK-8 (Sigma, 3 X 10"9 M). After obtaining a stable response, the tissues were washed out 3 times rapidly and every 5 to 10 minutes for 1 hour to reestablish a stable baseline.
  • Agonist EC50's Compounds were dissolved in dimethylsulfoxide (DMSO) then diluted with water and assayed via a cumulative concentration-response curve to test compound (10" ' ' 1 to 3 X 10 ⁇ 5 M) followed by a concentration-response curve to sulfated CCK-8 (10" ' ' 0 to 10"6 M) in the presence of the highest concentration of the test compound. As a final test, acetylcholine (1 mM) was added to induce maximal contraction. A minimum of three determinations of activity were made for each test compound.
  • DMSO dimethylsulfoxide
  • the cDNA clones for the human CCK-A 1 8 or CCK-B 19 receptors were ligated into pcDNA1-Neo vector from Invitrogen Corp (San Diego, CA) for direct transfection. DNA was prepared by the alkaline lysis method and transfected into CHO-K1 cells (ATCC,
  • Stable transfectants were initially selected by the use of Geneticin (Gibco BRL) and receptor bearing resistant cells were enriched by fluorescence-activated cell sorting based on binding of Fluorescein-Gly-[(Nle28,31]-CCK-8. Clonal lines were subsequently established by the limiting dilution method.
  • CCK-A or CCK-B receptor cDNA were grown at 37°C under a humidified atmosphere (95% 02 5% CO2) in Ham's F12 medium supplemented with 5% heat inactivated fetal bovine serum. The cells were passaged twice weekly and grown to a density of 2-4 million cells/mL. The cells were collected by centrifugation (600 X g, 15 min, 4°C) and resuspended in buffer (20 mL, pH 7.4) containing TrisHCI (25 mM), EDTA (5 mM), EGTA (5 mM), phenyl sulfonyl fluoride (0.1 mM) and soybean trypsin inhibitor (100 ⁇ g/mL).
  • Receptor Binding Assays 125 l-Bolton Hunter CCK-8 (Amersham, 2000 Ci/mmol) was dissolved in binding buffer (pH 7.4, 100,000 cpm/25 ⁇ L) containing HEPES (20 mM), NaCI (118 mM), KCI (5 mM), MgCI2 (5 mM) and EGTA (1 mM). Nonspecific binding was determined with MK-329 20 (10 ⁇ M, CCK-A) or L- 365,260 21 (10 ⁇ M, CCK-B). Test compounds were dissolved in DMSO at a stock concentration of 100 times the final assay concentration and diluted to appropriate concentrations with binding buffer.
  • Binding assays were performed in triplicate using 96-well plates to which the following were added sequentially: test compound (25 ⁇ L), 125 l-Bolton Hunter CCK-8 (25 ⁇ L), buffer (pH 7.4, 150 ⁇ L) and receptor preparation (50 ⁇ L). The final concentration of DMSO was 1 % in all assay wells. After 3 hours at 30°C, the incubation was terminated by rapid filtration of the mixture onto glass filters (Whatman GF/B) with subsequent washing to remove unbound ligand. Bound radioactivity was quantified by gamma counting.
  • the maximal response was normalized to the maximal response induced by CCK-8. ECso's were calculated at the concentration required to induce half-maximal response.
  • Anorexia Assays Male Long-Evans rats (225-300 g) were conditioned for two weeks to consume a palatable liquid diet (Bio-Serve F1657, Frenchtown, NJ) after a 2 hr fast. On pretreatment day, rats were fasted (100 min) and injected IP with drug vehicle (propylene glycol, PG, 1 mlJkg) and an oral preload of saline (0.9% NaCI, 8 mL/kg). Liquid diet access was provided 20 min later and consumption was measured at 30, 90 and 180 min. To qualify for the drug treatment study, rats had to consume at least 8 mL of liquid diet within the first 30 minutes on the pretreatment day.
  • drug vehicle propylene glycol, PG, 1 mlJkg
  • rats (8 - 10 animals per dose) were treated IP or PO with vehicle (PG, 1 mL/kg) or various doses (0.01 to 10 ⁇ mol/kg) of test compound dissolved in PG (1 mLJkg), immediately followed by the saline oral preload.
  • Food access was again provided 20 min later and food intake was measured at 30, 90 and 180 min. All food intake data were normalized for each rat to the respective values from the pretreatment day. Potency was determined at 30 min and efficacy at the 30 min, 1 ⁇ mol/kg dose.
  • the (+) enantiomer was four-fold more potent than the racemate in the in vitro isolated guinea pig gallbladder test ("GPGB").
  • the (+) enantiomer was eight-fold more potent than the racemate in the mouse gallbladder emptying assay (oral dosing). This increased potency is expected to be beneficial in the treatment of gallbladder stasis and in the treatment of obesity, since gallbladder stasis is a critical problem with rapid weight loss.
  • Anorectic agents are intended for chronic use and thus it is essential that they possess minimal risk for toxicity.
  • the primary toxicity associated with the use of cholecystokinin is concomitant CCK-B receptor agonist activity.
  • Activation of the CCK-B receptor is primarily associated with increased anxiety and increased gastric acid secretion.
  • CCK-B antagonists have been explored for both the development of anxiolytic agents and anti-ulcer agents. See, for example, Lowe, J, "Cholecystokinin-B Receptor Antagonists" in Exp. Opin. Ther. Patents, 5(3), pp 231-237 (1995).
  • the predominant CCK receptor subtype in the rodent pancreas is the CCK-A subtype and activation of this subtype induces pancreatic hyperstimulation and hypertrophy in rodents. Both of these activities are considered to be undesirable.
  • the tissue distribution of CCK receptors in human tissues has been reported.
  • the predominate receptor subtype in human pancreas is the CCK-B receptor subtype. See, for example, Wank, S. A., "Cholecystokinin Receptors" in American Journal of Physiology - Gastrointestinal & Liver Physiology, 32(5):, pp G628-G646, (1995).
  • activation of the CCK-B receptor could induce increased anxiety and gastric acid secretion, as well as pancreatic hyperstimulation and hypertrophy with long term use.
  • the preferred compound should have affinity for the CCK-B receptor and have measurable CCK-B antagonist activity in in vitro assays. Both enantiomers and the racemate are CCK-B antagonists. Although all three compositions have similar human CCK-A receptor affinities (IC50) and efficacies (EC50), the (+) enantiomer has the highest CCK-B receptor affinity (IC50) and selectivity (46- fold). Thus, the (+) enantiomer is preferred both in terms of CCK-A potency and efficacy, as well as in terms of the minimal potential for CCK-B induced toxic side effects.
  • the (+) enantiomer of the invention is essentially non-toxic at therapeutically useful doses.
  • the maximum non-lethal dose was found to be greater than 2000mg/kg in the rat and 1000mg/kg for male mice and 500mg/kg for female mice.

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
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  • Pharmacology & Pharmacy (AREA)
  • Medicinal Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
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  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Diabetes (AREA)
  • Hematology (AREA)
  • Obesity (AREA)
  • Child & Adolescent Psychology (AREA)
  • Emergency Medicine (AREA)
  • Endocrinology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Epidemiology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Nitrogen Condensed Heterocyclic Rings (AREA)
  • Plural Heterocyclic Compounds (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)

Abstract

La présente invention concerne un composé énantiomériquement enrichi représenté par la formule (I), des processus permettant de le préparer, des compositions pharmaceutiques le contenant et l'utilisation de celles-ci, dans le cadre du traitement des maladies induites par CCK-A ou d'états, tels que l'obésité.
EP00929488A 1999-05-06 2000-05-04 Derives de 1,5-benzodiazepines Withdrawn EP1212305A2 (fr)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
GBGB9910366.5A GB9910366D0 (en) 1999-05-06 1999-05-06 1,5-Benzodiazepine Derivatives
GB9910366 1999-05-06
GBGB0008179.4A GB0008179D0 (en) 2000-04-05 2000-04-05 Chemical process
GB0008179 2000-04-05
PCT/EP2000/003982 WO2000068209A2 (fr) 1999-05-06 2000-05-04 Derives de 1,5-benzodiazepines

Publications (1)

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EP1212305A2 true EP1212305A2 (fr) 2002-06-12

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JP (1) JP2002544200A (fr)
KR (1) KR20020012567A (fr)
CN (1) CN1161345C (fr)
AR (1) AR022044A1 (fr)
AU (1) AU4754800A (fr)
BR (1) BR0010338A (fr)
CA (1) CA2370801A1 (fr)
CO (1) CO5170461A1 (fr)
CZ (1) CZ20013988A3 (fr)
HK (1) HK1046138A1 (fr)
HU (1) HUP0201181A3 (fr)
IL (1) IL146124A0 (fr)
MX (1) MXPA01011267A (fr)
MY (1) MY138319A (fr)
NO (1) NO20015397D0 (fr)
NZ (1) NZ515048A (fr)
PE (1) PE20010110A1 (fr)
PL (1) PL351546A1 (fr)
TR (1) TR200103169T2 (fr)
WO (1) WO2000068209A2 (fr)

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WO2005035793A2 (fr) * 2003-10-09 2005-04-21 Decode Genetics Ehf. Marqueurs et haplotypes de cckar associes a des etats d'excedent de poids
JP4069159B2 (ja) 2004-05-25 2008-04-02 ファイザー・プロダクツ・インク テトラアザベンゾ［ｅ］アズレン誘導体及びそれらのアナログ
BR112022006546A2 (pt) 2019-10-07 2022-08-30 Kallyope Inc Agonistas de gpr119

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GB9307833D0 (en) * 1993-04-15 1993-06-02 Glaxo Inc Modulators of cholecystokinin and gastrin

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JP2002544200A (ja) 2002-12-24
CN1161345C (zh) 2004-08-11
KR20020012567A (ko) 2002-02-16
WO2000068209A3 (fr) 2001-03-01
TR200103169T2 (tr) 2002-04-22
PE20010110A1 (es) 2001-03-05
BR0010338A (pt) 2002-02-13
IL146124A0 (en) 2002-07-25
NO20015397L (no) 2001-11-05
HUP0201181A2 (hu) 2002-12-28
CZ20013988A3 (cs) 2002-02-13
AR022044A1 (es) 2002-09-04
CN1362949A (zh) 2002-08-07
HUP0201181A3 (en) 2003-10-28
PL351546A1 (en) 2003-05-05
NO20015397D0 (no) 2001-11-05
MXPA01011267A (es) 2002-05-06
MY138319A (en) 2009-05-29
AU4754800A (en) 2000-11-21
NZ515048A (en) 2004-01-30
HK1046138A1 (zh) 2002-12-27
CO5170461A1 (es) 2002-06-27
WO2000068209A2 (fr) 2000-11-16
CA2370801A1 (fr) 2000-11-16

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