GB2271991A - N-(2-oxo-1H-1,4-benzodiazepin-3-yl)-ureas - Google Patents

N-(2-oxo-1H-1,4-benzodiazepin-3-yl)-ureas Download PDF

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Publication number
GB2271991A
GB2271991A GB9322017A GB9322017A GB2271991A GB 2271991 A GB2271991 A GB 2271991A GB 9322017 A GB9322017 A GB 9322017A GB 9322017 A GB9322017 A GB 9322017A GB 2271991 A GB2271991 A GB 2271991A
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Prior art keywords
benzodiazepin
dihydro
phenyl
methyl
urea
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GB9322017D0 (en
Inventor
Mark Stuart Chambers
Stephen Robert Fletcher
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Organon Pharma UK Ltd
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Merck Sharp and Dohme Ltd
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Priority claimed from GB929222934A external-priority patent/GB9222934D0/en
Priority claimed from GB929223584A external-priority patent/GB9223584D0/en
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Priority to GB9322017A priority Critical patent/GB2271991A/en
Publication of GB9322017D0 publication Critical patent/GB9322017D0/en
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    • 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
    • C07D243/14—1,4-Benzodiazepines; Hydrogenated 1,4-benzodiazepines

Abstract

Compounds of the formula (I): <IMAGE> wherein R1 is hydrogen or specified optionally substituted alkyl, R<2> is specified optionally substituted phenyl or pyridyl, x is 0, 1, 2 or 3, R<3> is a specified alkyl, halo or amino and R<4> is specified cycloalkyl or bicycloalkyl; and salts and prodrugs thereof, are useful as antagonists of cholecystokin and gastrin receptors.

Description

BENZODIAZEPINE DERIVATIVE8 This invention relates to benzodiazepine compounds which are useful as antagonists of cholecystokinin and gastrin receptors.
Cholecystokinins (CCK) and gastrin are structurally related peptides which exist in gastrointestinal tissue and in the central nervous system (see, V. Mutt, Gastrointestinal Hormones, G.B.J. Green, Ed., Raven Press, N.Y., p.169 and G. Nission,- ibid.
p.127).
Cholecystokinins include CCK-33 , a a neuropeptide of thirty-three amino acids in its originally isolated form (see, Mutt and Jorpes, Biochem. J. 125, 678 (1971)), its carboxylterminal octapeptide, CCK-8 (also a naturally-occurring neuropeptide and the minimum fully active sequence), and 39- and 12-amino acid forms.
Gastrin occurs in 34-, 17- and 14-amino acid forms, with the minimum active sequence being the C-terminal tetrapeptide, Trp-Met-Asp-Phe-NH2, which is the common structural element shared by both CCK and gastrin.
CCKs are believed to be physiological satiety hormones, thereby possibly playing an important role in appetite regulation (G. P. Smith, Eating and Its Disorders, A. J. Stunkard and E. Stellar, Eds, Raven Press, New York, 1984, p. 67), as well as stimulating colonic motility, gall bladder contraction, pancreatic enzyme secretion and inhibiting gastric emptying. They reportedly co-exist with dopamine in certain mid-brain neurons and thus may also play a role in the functioning of dopaminergic systems in the brain, in addition to serving as neurotransmitters in their own right (see A.
J. Prange et al., "Peptides in the Central Nervous System", Ann. Rests. Med. Chem 17, 31, 33 (1982] and references cited therein; J. A. Williams, Biomed Res. 3 107 (1982]; and J.E. Morley, Life Sci. 30, 479 (1982]).
The primary role of gastrin, on the other hand, appears to be stimulation of the secretion of water and electrolytes from the stomach and, as such, is involved in control of gastric acid and pepsin secretion. Other physiological effects of gastrin then include increased mucosal blood flow and increased antral motility. Rat studies have shown that gastrin has a positive trophic effect on the gastric mucosa, as evidenced by increased DNA, RNA and protein synthesis.
- There are at least two subtypes of cholecystokinin receptors termed CCK-A and CCK-B (T.H.
Moran et al., "Two brain cholecystokinin receptors: implications for behavioural actions", Brain Res., 362, 175-79 [1986]). Both subtypes are found both in the periphery and in the central nervous system.
CCK and gastrin receptor antagonists have been disclosed for preventing and treating CCK-related and/or gastrin related disorders of the gastrointestinal (GI) and central nervous (CNS) systems of animals, especially mammals, and more especially those of humans. Just as there is some overlap in the biological activities of CCK and gastrin, antagonists also tend to have affinity for both CCK-B receptors and gastrin receptors. Other antagonists have activity at the CCK-A subtype.
Selective CCK antagonists are themselves useful in treating CCK-related disorders of appetite regulatory systems of animals as well as in potentiating and prolonging opiate-mediated analgesia (see P. L. Faris et al., Science 226, 1215 (1984)], thus having utility in the treatment of pain. CCK-B and CCK-A antagonists have also been shown to have a direct analgesic effect tM.F.
O'Neill et al., Brain Research, 534 287 (1990)).
Selective CCK and gastrin antagonists are useful in the modulation of behaviour mediated by dopaminergic and serotonergic neuronal systems and thus have utility in the treatment of schizophrenia and depression (Rasmussen et. al., 1991, Eur. J. Pharmacol., 209, 135-138; Woodruff et. al., 1991, Neuropeptides, 19, 45-46; Cervo et. al., 1988, Eur. J. Pharmacol., 158, 53-59), as a palliative for gastrointestinal neoplasms, and in the treatment and prevention of gastrin-related disorders of the gastrointestinal system in humans and animals, such as peptic ulcers, Zollinger-Ellison syndrome, antral G cell hyperplasia and other conditions inH which reduced gastrin activity is of therapeutic value,.see e.g. U.S.Patent 4,820,834. Certain CCK antagonists are useful anxiolytic agents and can be used in the treatment of panic and anxiety disorders.
CCK has been reported to evoke the release of stress hormones such as adrenocorticotrophic hormone, ss- endorphin, vasopressin and oxytocin, CCK may function as a mediator of responses to stress and as part of the arousal system. CCK-A receptors are now known to be present in a number of areas of the CNS and may be involved in modulating all of the above.
CCK may be involved in the regulation of stress and its relationship with drug abuse e.g. alleviation of the benzodiazepine withdrawal syndrome (Singh et. al., 1992, Br. J. Pharmacol., 105, 8-10) and neuroadaptive processes.
Since CCK and gastrin also have trophic effects on certain tumours [K. Okyama, Hokkaido J. Med. Sci., 206-216 (1985)), antagonists of CCK and gastrin are useful in treating these tumours [see, R.D. Beauchamp et al., Ann. Sura., 202, 203 (1985)].
In the light of discussion in C. Xu et al., Peptides, 8, 1987, 769-772, CCK antagonists may also be effective-in neuroprotection.
CCK receptor antagonists have been found to inhibit the contractile effects of CCK on iris sphincter and ciliary muscles of monkey and human eyes (Eur. J.
Pharmacol., 211(2), 183-187; A. Bill et al., Acta Physiol. Scand., 138, 479-485 (1990)), thus having utility in inducing miosis for therapeutic purposes.
A class of benzodiazepine antagonist compounds has been reported which binds selectively to brain CCK (CCK-B and CCK-A) and gastrin receptors (see M. Bock et al., J. Med Chem., 32, 13-16 (1989].
The present invention provides benzodiazepine compounds of formula (I):
wherein: R represents H, C1-6alkyl, C3-7cycloalkyl, cyclopropylmethyl, CH2CO2R5 (where R5 is C14alkyl), CH2CONR6R7 (where R6 and R7 each independently represents H or C1-4alkyl, or R6 and R7 together form a chain (CH2)p where p is 4 or 5), C1-6alkylNR8R9 or C1-6alkylCONR8R9 where R8 and R9 together with the nitrogen atom to which they are attached form a 5 to 8 membered non-aromatic ring system containing a second heteroatom selected from O, S or NR10, where R12 is H or C1-4alkyl;; R2 represents a phenyl or pyridyl group optionally substituted by one or more substituents selected from C1-6alkyl, halo, hydroxy, C14alkoxy, (CH2)q-tetrazolyl optionally substituted in the tetrazole ring by C1-4alkyl, (CH2)q-imidazolyl, (CH2)q-triazolyl (where q is 0, 1, 2 or 3), 5-hydroxy-4-pyrone, NR6R7, NR10COR5, NR10CONR10'R5 (where R10 and R10' are each independently H or C1-4alkyl and R5 is as previously defined), CONR6R7 (where R6 and R7 are as previously defined), SO(Cl~6alkyl), SO2(C1-6alkyl), trifluoromethyl, CONHSO2R, SO2NHCOR (where R is C1-6alkyl, optionally substituted aryl, 2,2-difluorocyclopropane or trifluoromethyl), S02NHR12 (where R12 is a nitrogen containing heterocycle), B(OH)2 or (CH2)qCO2H, where q is as previously defined; or R2 represents a group
where X1 represents CH or N; W represents CH2 or NR10, where R10 is as previously defined, and W1 represents CH2, or W and W1 each represent 0; or R2 represents phenyl substituted by a group
wherein x2 is O, S or NR10, where R10 is as previously defined;Z is a bond, 0 or S; m is 1, 2 or 3; n is 1, 2 or 3; and y is 0, 1, 2 or 3; Each R3 represents C1-6alkyl, halo or NR6R7, where R6 and R7 are as previously defined; R4 represents bridged C6~10bicycloalkyl or C37cyc1oalkyl optionally substituted by one or more C1-4alkyl groups; x is 0, 1, 2 or 3; with the proviso that when R1 is H, Cl,galkyl, C3~7cycloalkyl, cyclopropylmethyl, CH2C2R5 or CH2CONR6R7, R2 represents phenyl substituted by
and salts and prodrugs thereof.
It will be appreciated that formula (I) is intended to embrace all possible isomers, including optical isomers, and mixtures thereof, including racemates.
As used herein, the definition of each expression, when it occurs more than once in any structure, is intended to be independent of its definition elsewhere in the same structure.
The present invention includes within its scope prodrugs of the compounds of formula (I) above. In general, such prodrugs will be functional derivatives of the compounds of formula (I) which are readily convertible in vivo into the required compound of formula (I). Conventional procedures for the selection and preparation of suitable prodrug derivatives are described, for example, in "Design of Prodrugs", ed. H.
Bungaard, Elsevier, 1985.
Halo includes fluoro, chloro, bromo and iodo.
Preferably halo will be fluoro or chloro.
As used herein, unless otherwise indicated, alkyl means straight or branched chain saturated hydrocarbon.
Unless otherwise stated, aryl means optionally substituted carbocyclic or heterocylic aromatic groups, especially phenyl Heteroaryl means aromatic rings preferably having 5 or 6 ring atoms. and containing at least one atom selected from 0, S and N..
A first subgroup of compounds according to the invention is represented by compounds of formula (I), and salts and prodrugs thereof, wherein R1 represents C16alkyl, C37cycloa1ky1, cyclopropylmethyl, CH2C02R5 or CH2CoNR6R7 (where R5, R6 and R7 are as previously defined); R2 represents phenyl substituted by a group:
wherein X2, Z, m, n and y are as previously defined; and R4 represents C37cycloalkyl.
A second subgroup of compounds according to the invention is represented by compounds of formula (I), and salts and prodrugs thereof, wherein R1 represents C1-6alkylNR@R@ or C16alkylCONR8R9; R2 represents a phenyl group optionally substituted by one or more of C1-6alkyl, halo, hydroxy, C14alkoxy, (CH2)g-tetrazolyl optionally substituted in the tetrazole ring by C1-4alkyl, (CH2)q-imidazolyl, (CH2)q-triazolyl, 5hydroxy-4-pyrone, NR6R7, CONR6R7, NR10COR, NR10CONR10'R5, SO(C1-6alkyl), SO2(C1-6alkyl), trifluoromethyl, CONHSO2R1l, SO2NHCOR, SO2NHCOR1l, SO2NHR12, B(OH)2, (CH2)qC02H; or R2 represents a group
where W and W' are as previously defined.
In one preferred subgrup of compounds according to the invention, R1 represents C1-6alkyl, such as C1-4alkyl, for example, methyl, ethyl, n-propyl or i-butyl.
In another preferred subgroup of compounds of the invention, R1 represents C2-3alkylNR8R9, where the group NR8R9 preferably represents morpholinyl.
Suitable values for R11 include methyl, ethyl, i-propyl, t-butyl, phenyl and trifluoromethyl.
When R11 is optionally substituted aryl, this will preferably be optionally substituted phenyl.
Suitable substituents include C14alkyl, C14alkoxy, halo and trifluoromethyl. Preferred is unsubstituted phenyl or phenyl substituted by C1-6alkyl, for example, phenyl substituted by C1-6alkyl in the ortho position.
When R11 is C1-6alkyl, it will preferably represent C1-4alkyl. Particularly preferred are methyl and iso-propyl, especially iso-propyl.
When R2 is phenyl or pyridyl substituted by SO2NHR, suitable values of R include, for example, thiazole, thiadiazole and pyrazine.
Preferably q is zero.
When R2 represents optionally substituted phenyl or pyridyl, the substituents will preferably be selected from C1-galkyl, such as methyl and ethyl, halo, such as chloro, bromo, fluoro and iodo, and trifluoromethyl.
When R2 represents monosubstituted phenyl, the substituent- will preferably be located at the 3- or 4position of the phenyl ring, more preferably at the 3position. When R2 represents disubstituted phenyl the- substituents will preferably be located at the 3- and 4positions of the phenyl ring.
When R2 represents optionally substituted pyridyl it will preferably represent optionally substituted 3-pyridyl. When R2 represents monosubstituted 3-pyridyl, the substituent will preferably be located at the 5-position of the pyridyl ring.
When R2 represents a group
where X1 is CH the fused 5-membered ring will preferably be fused across the 3- and 4-positions of the phenyl ring and where X1 is N the 5-membered ring will preferably be fused across the 4- and 5-positions of the pyridyl ring.
Preferably W and W1 are CH2.
When R2 represents phenyl substituted by a group
the substituent will preferably be located at the 3- or 4- position of the phenyl ring, more preferably at the 3 -position.
Preferably~m is 1 or 2, more preferably 1.
Preferably n is 1 or 2, more preferably 1.
Preferably y is 0 or 1, more preferably 0.
Suitable values for X2 include 0, S, NH and NCH3.
Preferably Z represents a bond.
Suitable values for R3 include methyl and dimethylamino.
Preferably x is 0 or 1, more preferably 0.
When R4 represents C6-10bicycloalkyl, it will preferably contain 7, 8 or 9 carbon atoms, more preferably 7 carbon atoms. A suitable example of a C1-10bicycloalkyl substituent is:
When R4 represents C37cycloalkyl optionally substituted by one or more C1-4alkyl groups, it will preferably represent optionally substituted cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl, more preferably cyclohexyl or cycloheptyl optionally substituted by one or more C1-4alkyl groups. Preferably C1-4alkyl represents methyl.
Preferred are compounds wherein the C37cycloa1kyl group is unsubstituted.
A preferred subgroup of compounds according to the invention is represented by compounds of formula (Ia):
where in R20 is C47cycloalkyl; R21 is C1-4alkyl, preferably methyl, and R22 is H, or R21 and R22 together form a chain (CH2)3; n is 2 or 3; and salts and prodrugs thereof.
A further preferred subgroup of compounds according to the invention is represented by compounds of formula (Ib):
where in x2 is as defined for formula (I); R23 is-Cl~6alkyl, preferably C14alkyl;- R24 is cyclopentyl, cyclohexyl-or cycloheptyl, more preferably cyclohexyl or cycloheptyl; z is 0 or 1; and salts and prodrugs thereof.
Preferably the salts of the compounds of formula (I) are pharmaceutically acceptable, but nonpharmaceutically acceptable salts may be used for the preparation of pharmaceutically acceptable salts. The pharmaceutically acceptable salts of the compounds of formula (I) include the conventional non-toxic salts or the quaternary ammonium salts of the compounds from formula (I) formed, e.g., from non-toxic inorganic or organic salts.For example, such conventional non-toxic salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulphuric, sulphamic, phosphoric, nitric and the like; and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, steric, lactic, malic, tartaric, citric, ascorbic, palmoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulphanilic, 2-acetoxy benzoic, fumaric, toluenesulphonic, methanesulphonic, ethane disulphonic, oxalic and isothionic.
The salts of the present invention can be synthesized from the compound of formula (I) which contain a basic or acidic moiety by conventional chemical methods. Generally, the salts are prepared by reacting the free base or acid with stoichiometric amounts or with an excess of the desired salt-forming inorganic or organic acid or base in a suitable solvent or various combinations of solvents.
The-present invention also encompasses-a pharmaceutical composition comprising a compound of formula (I), or a salt or prodrug thereof and a pharmaceutically acceptable carrier or diluent.
The compounds of formula (I) and their salts and prodrugs, may be administered to animals, preferably to mammals, and most especially to a human subject either alone or, preferably, in combination with pharmaceutically acceptable carriers or diluents, optionally with known adjuvants, such as alum, in a pharmaceutical compostion, according to standard pharmaceutical practice. The compounds can be administered orally, parenterally, including by intravenous, intramuscular, intraperitoneal or subcutaneous administration, or topically.
For oral use of an antagonist of CCK, according to this invention, the selected compounds may be administered, for example, in the form of tablets or capsules, or as an aqueous solution or suspension. In the case of tablets for oral use, carriers which are commonly used include lactose and corn starch, and lubricating agents, such as magnesium stearate, are commonly added. For oral administration in capsule form, useful diluents include lactose and dried corn starch.
When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening and/or flavouring agents may be added.
For intramuscular, intraperitoneal, subcutaneous and intravenous use, sterile solutions of the active ingredient are usually prepared, and the pH of the solutions should be suitably adjusted and buffered.
For intravenous use, the total concentration of solutes should be controlled in order to render the preparation isbfonic,- -- For topical administration, a compound of formula (I) may be formulated as, for example, a suspension, lotion, cream or ointment.
For topical administration, pharmaceutically acceptable carriers are, for example, water, mixtures of water and water-miscible solvents such as lower alkanols or arylalkanols, vegetable oils, polyalkylene glycols, petroleum based jelly, ethyl cellulose, ethyl oleate, carboxymethylcellulose, polyvinylpyrrolidone, isopropyl myristate and other conventionally-employed non-toxic, pharmaceutically acceptable organic and inorganic carriers.The pharmaceutical preparation may also contain non-toxic auxiliary substances such as emulsifying, preserving, wetting agents, bodying agents and the like, as for example, polyethylene glycols 200, 300, 400 and 600, carbowaxes 1,000, 1,500, 4,000, 6,000 and 10,000, antibacterial components such as quaternary ammonium compounds, phenylmercuric salts known to have cold sterilizing properties and which are non-injurious in use, thimerosal, methyl and propyl paraben, benzyl alcohol, phenyl ethanol, buffering ingredients such as sodium chloride, sodium borate, sodium acetates, gluconate buffers, and other conventional ingredients such as sorbitan monolaurate, triethanolamine, oleate, polyoxyethylene sorbitan monopalmitylate, dioctyl sodium sulfosuccinate, monothioglycerol, thiosorbitol, ethylenediamine tetraacetic acid, and the like.
The compounds of formula (I) antagonise CCK and/or gastrin and are useful for the treatment and prevention of disorders including central nervous system disorders wherein CCK and/or gastrin may be involved.
Examples of such disease states include gastrointestinal diseases, including gastrointestinal ulcers, such as peptic and duodenal ulcers, Irritable bowel syndrome, gastroesophagenal reflux disease or excess pancreatic or gastrin secretion, acute pancreatitis, or utility disorders; central nervous system disorders, including central nervous system disorders caused by CCK interaction with dopamine, serotonin and other monoamine neurotransmitters, such as neuroleptic disorders, tardive dyskinesia, Parkinson's disease, psychosis or Gilles de la Tourette syndrome; depression, such as depression resulting from organic disease, secondary to stress associated with personal loss, or idiopathic depression; schizophrenia; disorders of appetite regulatory systems; Zollinger-Ellison syndrome, antral and cell hyperplasia, or pain.
The compounds of formula (I) are particularly useful in the treatment or prevention of neurological disorders involving anxiety disorders and panic disorders, wherein CCK and/or gastrin is involved.
Examples of such disorders include panic disorders, anxiety disorders, panic syndrome, anticipatory anxiety, phobic anxiety, panic anxiety, chronic anxiety and endogenous anxiety.
The compounds of formula (I) are also useful for directly inducing analgesia, opiate or non-opiate mediated, as well as anesthesia or loss of the sensation of pain.
The compounds of formula (I) may further be useful for preventing or treating the withdrawal response produced by chronic treatment or abuse of drugs or alcohol. Such drugs include, but are not limited to benzodiazepines, cocaine, alcohol and nicotine.
The compounds of formula (I) may further by useful in the treatment of stress and its relationship with drug abuse.
- The compounds of formula (I) may further be useful in the treatment of oncologic disorders wherein CCK may be involved. Examples of such oncologic disorders include small cell adenocarcinomas and primary tumours of the central nervous system glial and neuronal cells. Examples of such adenocarcinomas and tumours include, but are not limited to, tumours of the lower oesophagus, stomach, intestine, colon and lung, including small cell lung carcinoma.
The compounds of formula (I) may also be useful as neuroprotective agents, for example, in the treatment and/or prevention of neurodegenerative disorders arising as a consequence of such pathological conditions as stroke, hypoglycaemia, cerebral palsy, transient cerebral ischaemic attack, cerebral ischaemia during cardiac pulmonary surgery or cardiac arrest, perinatal asphyxia, epilepsy, Huntington's chorea, Alzheimer's disease, Amyotrophic Lateral Sclerosis, Parkinson's disease, Olivo-ponto-cerebellar atrophy, anoxia such as from drowning, spinal cord and head injury, and poisoning by neurotoxins, including environmental neurotoxins.
The compounds of formula (I) may further be used to induce miosis for therapeutic purposes after certain types of examination and intraocular surgery. An example of intraocular surgery would include cateract surgery with implantation of an artificial lens. The CCK antagonist compounds of this invention can be used to prevent miosis occuring in association with iritis, ureitis and trauma.
The present invention therefore provides a compound of formula (I) or a salt or prodrug thereof for use in the preparation of a medicament.
The present invention also provides a compound of formula (I) for use in therapy.
In a further or alternaiive embodiment thepresent invention provides a method for the treatment or prevention of a physiological disorder involving CCK and/or gastrin which method comprises administration to a patient in need thereof of a CCK and/or gastrin antagonising amount of a compound of formula (I).
When a compound according to formula (I) is used as an antagonist of CCK or gastrin in a human subject, the daily dosage will normally be determined by the prescibing physician with the dosage generally varying according to the age, weight, and response of the individual patient, as well as the severity of the patient's symptoms. However, in most instances, an effective daily dosage wll be in the range from about 0.OOsmg/kg to about 100mg/kg of body weight, and preferably, of from 0.05mg/kg to about 50mg/kg, such as from about 0.smg/kg to about 20mg/kg of body weight, administered in single or divided doses. In some cases, however, it may be necessary to use dosages outside these limits. For example, animal experiments have indicated that doses as low as lng may be effective.
In effective treatment of panic syndrome, panic disorder, anxiety disorder and the like, preferably about 0.05 mg/kg to about 0.5 mg/kg of CCK antagonist may be administered orally (p.o.), administered in single or divided doses per day (b.i.d.). Other routes of administration are also suitable.
For directly inducing analgesia, anaesthesia or loss of pain sensation, the effective dosage preferably ranges from about 100 ng/kg to about lmg/kg by systemic administration. Oral administration is an alternative route, as well as others.
In the treatment or irritable bowel syndrome, preferably about 0.1 to 10 mg/kg of CCK antagonist is administered orally (p..), administered in singe -or divided doses per day (b.i.d.). Other routes of administration are also suitable.
The use of a gastrin antagonist as a tumour palliative for gastrointestinal neoplasma with gastrin receptors, as a modulator of central nervous activity, treatment of Zollinger-Ellison syndrome, or in the treatment of peptic ulcer disease, an effective dosage of preferably about 0.1 to about 10 mg/kg administered oneto-four times daily is indicated.
For use as neuroprotective agents the effective dosage preferably ranges from about 0.5mg/kg to about 2Omg/kg.
Because these compounds antagonise the function of CCK in animals, they may also be used as feed additives to increase the food intake of animals in daily dosage of preferably about 0.05mg/kg to about 50mg/kg of body weight.
According to one general process (A), the compounds of formula (I) may be prepared by reaction of intermediates of formula (II) with compounds of formula (III)
wherein R, R, R , R4 and x are as defined for formula (I) above, one of R30 and R represents NH2 and the other of R30 and R31 represents -N=C=O.
-The reaction is conveniently effected in a suitable organic solvent, such as an ether, for example, tetrahydrofuran.
According to a second general process, (B), compounds of formula (I) may be prepared by reacting a compound of formula (VI)
wherein R1, R3, R4 and x are as defined for formula (I) and Y represents an activated carbamate, with an amine of formula (III) wherein R31 is NH2 (hereinafter intermediates (IIIA)), in the presence of a base.
An "activated carbamate" is a carbamate group which bears a substituent which activates the carbamate function to nucleophilic attack. Suitably Y may represent an appropriately substituted aryl carbamate of 0 formula Ar-O-C-NH, for example
Suitable bases for use in the reaction include tertiary amines, for example, triethylamine.
The reaction is conveniently effected in a suitable organic solvent, for example, dimethylformamide, at ambient or elevated temperature. Preferably the reaction is conducted at approximately 50"C.
Intermediates of formula (II) wherein R30 is NH2 (hereinafter intermediates (IIA)) may be prepared from compounds of formula (VIII)
wherein R3, R4 and x are as defined for formula (I) and G is a protecting group; by reaction with a reagent suitable to introduce the group R1, for example a halide of formula R1Hal where Hal represents halo such as chloro, bromo or iodo, in the presence of a base, such as an alkali metal hydride or an alkaline earth metal carbonate, for example sodium hydride or caesium carbonate; or a suitable dialkyl acetal of dimethyl formamide in a suitable organic solvent, e.g. toluene, followed by deprotection.
Compounds of formula (VIII) may be prepared from compounds of formula (IX)
wherein R3, R4 and x are as defined for formula (I) by a reaction sequence comprising: (i) reaction with a compound of formula (X)
wherein G is as defined above, in the presence of a base, such as a tertiary amine, for example triethylamine or Nmethyl morpholine, and a coupling reagent. Any of the coupling reagents commonly used in peptide synthesis are suitable, for example, 1,3-dicyclohexylcarbodiimide (DCC) isobutyl chloroformate or, preferably, bis(2-oxo-3oxazolidinyl)phosphonic chloride (BOP-C1).
(ii) Treatment with gaseous ammonia, preferably in the presence of a mercury containing catalyst, such as mercury(II) chloride. The reaction is conveniently effected in a suitable organic solvent, such as an ether, for example, tetrahydrofuran.
(iii) Treatment with an organic acid, for example acetic or propionic acid, optionally in the presence of an ammonium salt, for example ammonium acetate.
Compounds of formula (IX) may be prepared by reaction of a compound of formula (XI) with a compound of formula (XII)
wherein R3, R4 and x are as defined for formula (I), R40 and R41 each represent Cl,qalkyl, preferably dimethyl, and G' represents a protecting group, such as tbutoxycarbonyl group, in the presence of a base, followed by deprotection.
Suitable bases of use in the reaction include organolithiums, such as butyllithiums, for example t butyll ithium.
The reaction is conveniently effected in a suitable organic solvent, such as an ether, for example tetrahydrofuran, at low temperature.
Compounds of formula (XI) may be prepared from the corresponding acids of formula R4COOH, wherein R4 is as defined for formula (I), by reaction with an amine of formula R40NHR40, wherein R40 and R41 are as previously defined, in the presence of a base and a coupling agent.
Suitable bases of use in the reaction include tertiary amines such as, for example, triethylamine. The reaction is conveniently effected in a suitable organic solvent, such as dimethylformamide. A preferred coupling agent of use in the reaction is 1-(3dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC).
Compounds of formula (XII) and acids of formula R4COOH are commercially available or may be prepared from commercially available starting materials by conventioal methods-well known to those. skilled in the art.
Alternatively, compounds of- formula (IX) may be prepared by reaction of a compound of formula (XIV)
wherein R3 and x are as previously defined, with a Grignard reagent of formula R4MgHal wherein R4 is cycloalkyl and Hal is halo such as chloro, bromo or iodo.
Compounds of formula (XIV) are commercially available or may be prepared from commercially available compounds by conventional methods.
Intermediates of formula (II) wherein R30 is -N=C=O (IIB) may be prepared from amines of formula (II) wherein R30 is NH2 (IIA) by reaction with triphosgene in the presence of a base, such as a tertiary amine, for example, triethylamine. The reaction is conveniently effected in a suitable organic solvent, such as an ether, for example, tetrahydrofuran, suitably at low temperature, such as about 0 C.
Intermediates of formula (VI) may be prepared from compounds of formula (IIA) by reaction with a 0 suitable haloformate of formula Ar-O-C-Hal, where Hal is as previously defined, preferably chloro, for example
in the presence of a base, such as a tertiary amine, for example, triethylamine.
Isocyanates of formula (III) wherein R31 is -N=C=O (IIIB) may be prepared from amines of formula (IIIA) by conventional methods.
Amines of formula (IIIA) are known compounds, or may be prepared from the corresponding nitro compounds of formula R2NO2, wherein R2 is as previously defined, by reduction.
Suitably the reduction is effected by catalytic hydrogenation, for example, using a noble metal catalyst such as palladium which may be supported, e.g. on carbon.
The reaction is conveniently effected in a suitable organic solvent, such as an alcohol, e.g. ethanol.
Compounds of formula (XVI) are commercially available or may be prepared by the procedures described in the accompanying Examples or by alternative procedures which will be readily apparent to one skilled in the art.
Where the above-described process for the preparation of the compounds according to the invention gives rise to mixtures of stereoisomers these isomers may, if desired, be separated, suitably by conventional techniques such as preparative chromatography.
The novel compounds may be prepared in racemic form, or individual enantiomers may be prepared either by enantiospecific synthesis or by resolution. The novel compounds may, for example, be resolved into their component enantiomers by standard techniques, such as the formation of diastereomeric pairs by salt formation with an optically active acid, such as (-)-di-p-toluoyl-Ltartaric acid and/or (+)-di-p-toluoyl-D-tartaric acid followed by fractional crystallization and regeneration - of the free base. The novel compounds may also be resolved by - formation - of diastereomeric esters or - amides, followed by chromatographic separation and removal of the chiral auxiliary. Alternatively, enantiomers of the novel compounds may be separated by HPLC using a chiral column.
During any of the above synthetic sequences it may be necessary and/or desirable to protect sensitive or reactive groups on any of the molecules concerned. This may be achieved by means of conventional protecting groups, such as those described in Protective Groups in Organic Chemistrv, ed. J.F.W. McOmie, Plenum Press, 1973; and T.W. Greene and P.G.M. Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 1991. The protecting groups may be removed at a convenient subsequent stage using methods known from the art.
The following examples are provided to assist in a further understanding of the invention. Particular materials employed, species and conditions are intended to be further illustrative of the invention and not limitative of the scope thereof.
4B, 5 and 6 of International Patent Application PCT/GB92/01366 published as WO 93/02078. When tested for CCK Receptor Binding, Pancreas and Brain, by the method described in PCT/GB92/01366 (WO 93/02078) the compounds of the Examples displayed an 1C50 < 1mM at the CCK receptor.
EXAMPLE 1: N-[3(R,S)-5-Cycloheptyl-2,3-dihydro-1-(2-(N morpholino)ethvl)-2-oxo-1H-1,4-benzodiazepin-3-vll N -f3- methylphenyl] urea Step 1: 2-Aminophenylcycloheptylmethanone Over a period of 1h a solution of cycloheptyl bromide (37.8g) in diethyl ether (200ml) was added dropwise to a slurry of magnesium turnings (5.28g) and a crystal of iodine in diethyl ether (20ml) at reflux.The mixture was stirred for a further hour whereupon the Grignard solution was cannulated into a pressure equalising dropping funnel, attached to a three-necked round-bottomed - flask, which was uider an - atmosphere of nitrogen; A solution of 2-aminobenzonitrile (8.26g) in diethyl ether (200ml), at 0 C, was treated dropwise with the Grignard reagent prepared above, over a period of 30 min. Once the addition was complete, the mixture was warmed to room temperature and stirred for 16h under nitrogen. The solution was cooled to 0 C, quenched with 5N hydrochloric acid (45ml), and basified using solid sodium hydroxide (8.9g).The aqueous solution was extracted with ethyl acetate (2 x 100ml) and the combined organic layers were dried (Na2SO4) and evaporated. The residue was chromatographed on silica gel using 2:1 petrol:ethyl acetate as the eluant. This gave the title compound (8.2g) as a pale yellow oil. 1H NMR (250MHz, CDCl3) # 1.40-2.10 (12H, m), 3.34-3.50 (1H, m), 6.30 (2H, brs), 6.60-6.70 (2H, m), 7.20-7.30 (1H, m), 7.70-7.80 (1H, m). TLC (silica, petrol (60/80):ethyl acetate 2:1). Rf= 0.50.
Step 2: Cycloheptyl-2-(&alpha;-(benzyloxycarbonylamino)-&alpha;-iso- propvlthioacetvlamino)phenvl methanone a-(iso-Propylthio)-N-(benzyloxycarbonyl)glycine (21.9g) was dissolved in dichloromethane (450ml) and cooled to 0 C.
The stirred solution was then treated with triethylamine (21.5ml), bis(2-oxo-3-oxazolidinyl)phosphinic chloride (19.7g) and 2-nminophenyl cycloheptyl methanone (12.0g). The mixture was warmed to ambient temperature and stirred for 2h. The mixture was then washed in succession with 10% citric acid solution (1OOml), water (100ml), saturated sodium bicarbonate solution (100ml) and brine (100ml). The dried (Na2SO4) organic phase was evaporated and the residue chromatographed on silica gel using 8:1 petrol:ethyl acetate as eluant.This afforded the title compound as a colourless solid (13.5g). 1H NMR (250MHz, CDC13) 1.10-2.00 (18, m), 3.14-3.34 (1H, m), 3.40-3.56 (1H, m), 5.16-5.22 (2H, m), 5.56 (1H, d, J=5Hz), 5.98 (1Htd, J=5Hz), 7.10-7.18 (1H, m), 7.24-7.44 (5H, m), 7.50-7.60.
(1H, m), 7.84-i.94 (1H, m), 8.60-8.70 (1H, m), 12.28 (1H, brs).
TLC (silica, petrol (60/80):ethyl acetate 3:1). Rf = 0.45.
Step 3: 3(R.S)-r(Benzvlogvcarbonsl)ninol-5-cvcloheptvl- 1,3-dihydro-2H-1,4-benzodiazepin-2-one Cycloheptyl-2-(&alpha;-(benzyloxycarbonylamino)-&alpha;-iso- propylthioacetylamino)phenyl methanone (lOg) was dissolved in anhydrous tetrahydrofuran (500ml) and cooled to 0 C. Ammonia gas was bubbled through the stirred solution for 10 min before adding mercuric chloride (8.5g) in one portion. Ammonia was continually bubbled through the solution for a further hour, then the suspended solids were filtered off. The solvent was evaporated in vacuo to leave an oil, which was used without further purification.
The crude a-aminoglycinamide was dissolved in glacial acetic acid (200ml) and treated with ammonium acetate (7.7g).
The resulting reaction mixture was stirred at room temperature overnight, before removing the solvent in vacuo. The residue was partitioned between ethyl acetate (300ml) and 10% sodium hydroxide solution (200ml). The dried (Na2SO4) organic layer was evaporated and the residue chromatographed on silica gel with 2:1 petrol:ethyl acetate as eluant. This afforded the title product (8.0g) as a colourless solid. 1H NMR (250MHz, CDCl3) 3 1.18-2.20 (12H, m), 2.90-3.07 (1H, m), 5.06-5.24 (3H, m), 6.46 (1H, d, J=1OHz), 7.04-7.12 (1H, m), 7.22-7.42 (6H, m), 7.44-7.56 (1H, m), 7.58-7.68 (1H, m), 10.30 (1H, brs). TLC (silica, petrol (60/80):ethyl acetate 2:1). Rf= 0.15.
Step 4: 3(R.S)-f(Benzvloxvcarbonvl)aminol-5-cveloheptvl- 1.3-dihvdro- 1-(2-(N-morpholino)ethvl)-2H- 1.4-benzodiazepin-2-one 3(R,S)-[(Benzyloxycarbonyl)amino]-5-cycloheptyl-1,3-dihydro 2H-1,4-benzodiazepin-2-one (0.5g) in anhydrous dimethylformamide (20ml) was treated with cesium carbonate (1.2g) and N-(2chloroethyl)morpholine (0.6g). The mixture was stirred at 60 C for 16h. The undissolved material was removed by filtration. The solvent was evaporated and the residue partitioned between ethyl acetate (50ml) and water (30ml). The organic phase was separated, dried (Na2SO4) and evaporated in vacuo.The residue was triturated with diethyl ether to afford the title compound (0.43g) as a colourless solid, mp 130-20C. 1H NMR (250 MHz, D6-DMSO) 3 1.14-1.86 (11H, m), 1.92-2.08 (1H, m), 2.10-2.38 (6H, m), 3.04-3.20 (1H, m), 3.36-3.54 (4H, m), 3.76-3.92 (1H, m), 4.24-4.40 (1H, m), 4.88 (1H, d, J=8.3Hz), 5.00, 2H, s), 7.14-7.40 (5H, s), 7.54-7.80 (3H, m), 8.20 (1H, d, J=8.3Hz).
Step 5: 3(R.S)-Amino-5-cvdohentvl- 1 .3-dihvdro-1-(2-N- morpholino)ethvl)-2H-1.4-benzodiazepin-2-one 3(R,S)-[(Benzyloxycarbonyl)amino]-5-cycloheptyl-1,3 dihydro- 1-(2-(N-morpholino)ethyl)-2H- 1,4-benzodiazepin-2-one (300mg) was treated with a solution of 30% hydrogen bromide in glacial acetic acid (15ml) and stirred for 20 min at room temperature. The mixture was then added dropwise onto cold (0 C) diethyl ether (50ml). A colourless solid precipitated which was filtered off. The solid was treated with 10% sodium hydroxide solution (50ml) then extracted with ethyl acetate (80ml). The organic layer was separated, dried (Na2SO4) and evaporated to afford the title compound (218mg) as a colourless oil.TLC (silica/dichloromethane:methanol 9:1) Rf, 0.40. 1H NMR (250 MHz, CDCl3) o: 1.26-1.74 (9H, m), 1.76-1.96 (2H, m), 2.00-2.16 (1H, m), 2.30-2.60 (8H, m), 2.88-3.00 (1H, m), 3.50-3.70 (4H, m), 3.72-3.86 (1H, m), 4.30-4.42 (1H, m), 7.20-7.30 (1H, m), 7.40-7.60 (3H, m).
Step 6: N-[3(R,S)-5-Cycloheptyl-2,3-dihydro-1-(2-(Nmorpholino)ethyl)-2-oxo-1H-1,4-benzodiazepin-3-1]N'-3methylphenyl]urea 3(R, S)-Amino-5-cycloheptyl- 1,3-dihydro- 1-(2-(N- morpholino)ethyl)-2H-1,4-benzodiazepin-2-one (214mg) in dry tetrahydrofuran (5ml) was treated with 3-methylphenyl isocyanate (72p1). The mixture was stirred for 30 min.The solvent was evaporated and the residue triturated with diethyl ether to afford the. title compound. (100mg) as a colourless solid, mp 206-2070C. 1H NMR (360 MHz, D6-DMSO)~ #1.14-1.80-(11H, m), 1.95-2.06 (1H, m), 2.10-2.37 (9H, m), 3.08-3.19 (1H, m), 3.36-3.50 (4H, m), 3.80-3.93 (1H, m), 4.25-4.35 (1H, m), 5.00 (1H, d, J=8.4Hz), 6.71 (1H, d, J=6.6Hz), 7.04-7.14 (2H, m), 7.17 (1H, s), 7.25 (1H, d, J=8.5Hz), 7.37 (1H, t, J=7.1Hz), 7.61 (1H, t, J=8.5Hz), 7.67-7.82 (2H, m), 8.86 (1H, s).
EXAMPLE 2: N-TSR. $)-5-Cvlohexvl-2 .3-dihvdro- 1-(2-(N- morpholino)ethvl)-2-oxo-lH-1.4-benzodiazepin-3-vllN'-r3- methvlphenvll urea Step 1: (2-Acetamidophenvl) cvelohexvl methanone Cyclohexylmagnesium bromide (240ml of a 2M solution in ether) in ether (200ml) was added dropwise to a solution of 2-methyl-4H-3,1-benzoxazin-4-one (100g) in ether (1100ml) at -10 C over 2h. The mixture was stirred at this temperature for 2h, then at ambient temperature for 30 min.After cooling to -10 C the suspension was treated with 2M HCl (600ml), keeping the temperature below 000. After stirring for 15 min the layers were separated, and the ethereal layer washed sequentially with water (500ml), 5% sodium hydroxide solution (2 x 500ml) and finally water (2 x 500ml). The organic layer was separated, dried (MgSO4), evaporated in vacuo and chromatographed on silica gel using petrol:ethyl acetate (2:1) to give (2-acetamidophenyl) cyclohexyl methanone (28g). Mp 66 C. 1H NMR (CDCl3, 360MHz) 8 1.25-1.89 (10H, m), 2.23 (3H, s), 3.33 (1H, m), 7.13 (1H, d oft, J=6 and 1Hz), 7.53 (1H, d oft, J=6 and 1Hz), 7.92 (1H, d, J=6Hz), 8.76 (1H, d, J=6Hz), 11.73 (1H, brs).
Step 2: (2-Aminophenvl) cvclohexvl methanone A solution of (2-acetamidophenyl) cyclohexyl methanone (0.53g) in methanol (5ml) and concentrated hydrochloric acid (15ml) was heated at 800C for lh. After this time the solution was cooled to ambient temperature-and the solvents removed in vacuo. The residue was dissolved in water (10ml) and basified with 4N sodium hydroxide solution (20ml). The mixture was then extracted into ethyl acetate (4 x 20ml) and the organic layers combined and dried (MgSO4).The solvent was evaporated and the residue chromatographed on silica gel using petrol:ethyl acetate (2:1), to afford the amine (0.40g). Mp 73-75 C. 1H NMR (360MHz, CDCl3) 1.23-2.09 (10H, m), 3.27 (1H, m), 6.29 (2H, brs), 6.64 (2H, m), 7.25 (1H, dt, J=6 and 1Hz), 7.76 (1H, dd, J=7 and 1Hz).
Step 3: 3(R.S)-T(Benzvl oxvcarbonvl )aminol-5-cvclohexvl- 1,3-dihydro-2H- 1.4-benzodiazepin-2-one a-(Isopropylthio)-N-(benzyloxycarbonyl)glycine (30g) was dissolved in dichloromethane (1000ml) and cooled to 05C. The stirred solution was then treated with N-methyl morpholine (11.5ml) followed by isobutyl chloroformate (13.7ml). The resulting reaction mixture was stirred for a further 15 min at OOC, then heated to reflux. The refluxing reaction mixture was treated dropwise, over 20 min, with a solution of (2-aminophenyl) cyclohexyl methanone (20.5g) in dichloromethane (140ml). After addition was complete the reaction was heated at reflux for a further 4h.The mixture was then washed in succession with 10% citric acid solution (2 x 500ml), saturated sodium bicarbonate solution (2 x 500ml) and brine (500ml). The dried (MgSO4) organic phase was evaporated to afford the crude product as a pale orange solid, which was used without further purification.
The crude (isopropylthio)glycinamide was dissolved in anhydrous tetrahydrofuran (800ml) and cooled to 0 C. Ammonia gas was bubbled through the stirred solution for 30 min before adding mercuric chloride (33g) in one portion. Ammonia was continually bubbled through the solution for a further 5 hours, then the suspended solids were filtered off. The solvent was evaporated in vacuo.
The crude a-aminoglycinamide was dissolved in glacial acetic acid (500ml) and treated with ammonium acetate (36.2g). The resulting reaction mixture was stirred at room temperature overnight, before removing the solvent in vacuo. The residue was partitioned between ethyl acetate (300ml) and 1N sodium hydroxide solution (300ml). The organic phase was separated, dried (MgSO4) and evaporated. The residue was chromatographed on silica gel, using 2:1 petrol:ethyl acetate as the eluant, to afford 3(R,S) t(benzYloxycarbonyl)smino]-5- cyclohexyl-1,3-dihydro-2H-1,4- benzodiazepin-2-one (25g). Mp 164-166 C. 1H NMR (360MHz, CD(:13) 61.07-2.04 (10H, m), 2.77 (1H, m), 5.12 (3H, m), 6.44 (1H, d, J=8Hz), 7.08 (1H, d, J=8Hz), 7.23-7.36 (6H, m), 7.46 (1H, t, J=7Hz), 7.59 (1H, d, J=8Hz), 8.60 (1H, brs).
Step 4: 3(R,S)-[(Benzyloxycarbonyl)amino]-5-cyclohexyl- 1.3 dihvdro- 1-(N-(2-ethvl)morpholino)-2H- 1 .4-benzodiazepin-2- one Prepared from 3 (R,S)-C(B enzyloxycarbonyl)amino] -5 -cyclohexyl- 1,3-dihydro-2H-1,4-benzodiazepin-2-one (1.0g) by the method of Example 1, Step 4. TLC (silica/ dichloromethane:methanol 19:1) Rf, 0.27. 1H NMR (250 MHz, D6-DMSO) (o: 0.90-1.69 (8H, m), 1.71-1.89 (1H, m), 1.86-2.00 (1H, m), 2.10-2.34 (6H, m), 2.84-3.00 (1H, m), 3.35-3.50 (4H, m), 3.75-3.90 (1H, m), 4.26-4.40 (1H, m), 4.92 (1H, d, J=7.5Hz), 5.03 (2H, s), 7.22-7.43 (5H, s), 7.54-7.80 (3H, s), 8.22 (1H, d, J=7.5Hz).
Step 5: 3(R,S)-Amino-5-cyclohexyl-1,3-dihydro-1-(2-(Nmorpholino)ethyl)-2H-1,4-benzodiazepin-2-one Prepared from 3(R,S)-[(benzyloxycarbonyl)amino]-5cyclohexyl-1,3-dihydro-1-(2-N-morpholino)ethyl)-2H-1,4benzodiazepin-2-one (0.86g) by the method of Example 1, Step 5.
TLC (silica/dichloromethane:methanol 9:1) Rf, 0.26. 1H NMR (250MHz, D6-DMSO) #0.86-1.70 (8H, m), 1.73-1.86 (1H, m), 1.88-2.00 (1H, m), 2.12-2.30 (6H, m), 2.80-2.94 (1H, m), 3.08 (2H, brs), 3.38-3.46 (4H,-m), 3.76-3.88(1H, m), 4.22-4.36 (1H, m), 7.12-7.36 (2H, m), 7.52-7.70 (2H, m).
Step 6: N-[3(R,S)-5-Cyclohexyl-2,3-dihydro-1-(2-(Nmorpholino)ethyl)-2-oxo-1H-1,4-benzodiazepin-3-yl]N'-[3 methvlphenvll urea Prepared from 3(R,S)-Amino-5-cyclohexyl-1,3-dihydro-1 (2-(N-morpholino)ethyl)-2H-1,4-benzodiazepin-2-one (300mg) by the method of Example 1, Step 6. Mp 216.5-218.5 C. 1H NMR (360 MHz, D6-DMSO) 3 1.00-1.70 (8H, m), 1.74-1.84 (1H, m), 1.90-2.00 (1H, m), 2.14-2.36 (9H, m), 2.90-3.01 (1H, m), 3.36-3.50 (4H, m), 3.82-3.90 (1H, m), 4.30-4.39 (1H, m), 5-03 (1H, d, J=8.5Hz), 6.71 (1H, d, J=7.2Hz), 7.05-7.14 (2H, m), 7.17 (1H, s), 7.26 (1H, d, J=8.5Hz), 7.37 (1H, t, J=7.4Hz), 7.61 (1H, t, J=7.2Hz), 7.65-7.80 (2H, m), 8.87 (1H, s).
EXAMPLE 3: N-[3(R,S)-5-Cyclohexyl-2,3-dihydro-1-(2-(Nmorpholino)ethyl)-2-oxo-1H-1,4-benzodiazepin-3-yl]N'-(5 indanvliurea 5-Aminoindane (163mg) in dry tetrahydrofuran (40ml) was cooled to OOC whereupon triphosgene (121mg) was added. The mixture was stirred at OOC for 2 min, then triethylamine (513 l) was added portionwise until pH8 was attained. The mixture was then stirred for a further 5 min, allowed to warm to 1500 and then recooled to 05C. Then a solution of 3(R,S)-amino-5-cyclohexyl-1,3dihydro-1-(2-N-morpholino)ethyl)-2H-1,4-benzodiazepin-2-one example 2, Step 5] (300mg) in anhydrous tetrahydrofuran (5ml) was added dropwise over 5 min.The mixture was stirred at OoC-for 5 min, allowed to warm to room temperature and then stirred for a further 30 min. The undissolved material was removed by filtration.
The solvent was evaporated in vacuo and the residue partitioned between ethyl acetate (50ml) and water (20ml). The organic phase was separated, dried (Na2SO4) and evaporated. The crude solid was chromatographed on silica gel with a gradient elution of 0-3% methanol in dichloromethane to afford the title compound (114mg), mp 247 C (dec.). 1H NMR (360 MHz, D6-DMSO) 80.90-1.70 (10H, m), 1.75-1.84 (1H, m), 1.90-2.02 (3H, m), 2.15-2.55 (6H, m), 2.70-2,82 (4H, m), 2.95-3.00 (1H, m), 3.40-3.50 (4H, m), 3.80-3.92 (1H, m), 4.30-4.40 (1H, m), 5.03 (1H, d, J=8.5Hz), 7.02-7.04 (2H, m), 7.20 (1H, d, J=8.5Hz). 7.24 (1H, s), 7.37 (1H, t, J=7.4Hz), 7.61 (1H, t, J=7.2Hz), 7.70-8.02 (2H, m), 8.80 (1H, s).
EXAMPLE 4: N-[3(R,S)5-Cyclohexyl-2 .3-dihvdro-2-oxo- 1-nropvl- 1H-1,4-benzodiazepin-3-yl]N'-[4-(N-piperazinyl)phenyl]urea, hvdrochloride salt Step 1: 3(R.S)-r(Benzvloxvcarbonvl)aminol-5-cvelohexvl-1.3- dihydro-1-propyl-2H-1,4-benzodiazepin-2-one A solution of 3(R,S)-[(benzyloxycarbonyl)amino]-5- cyclohexyl1,3-dihydro-2H-1,4-benzodiazepin-2-one (Example 2, Step 3, 2g) in anhydrous dimethylformamide (15ml), under an atmosphere of nitrogen, was treated with sodium hydride (0.22g of a 55-60% dispersion in mineral oil, 5.lmmol) in one portion, at 000. After 45 min at 0 C, 1-iodopropane (0.55ml) was added in one portion and the solution allowed to reach ambient temperature and stirred overnight.
Solvent was removed under reduced pressure, and the crude residue partitioned between water (25ml) and dichloromethane (25ml). The organic phase was separated and the aqueous phase extracted with dichloromethane (3 x 25ml). The combined organic layers were washed with brine (50ml), dried (MgSO4) and evaporated in vacuo.
The residue was triturated with diethyl ether give the title compound (1.74g). Mp 160-163 C. 1H NMR (360MHz, CDCl3) 0.82 (3H, t, J=10.SHz), 0.94-1.48-(5H, m)j 1.50-1.76 (5H, m), 1.79-1.90 (1H, m), 1.96-2.08 (1H, m), 2.70-2.84 (1H, m),3.46-3.59 (1H, m), 4.22-4.35 (1H, m), 5.06-5.16 (3H, m), 5.07 (1H, d, J=12.0Hz), 7.21-7.40 (7H, m), 7.44-7.59 (2H, m).
Step 2: 5-Cvclohexvl-1.3-dihvdro-3-(R.S)-T(4- nitrophenyloxycarbonyl)amino]-1-propyl-2H-1,4-benzodiazepin-2-one 3(R,S)-t(Benzyloxycarbonyl)amino]-5-cyclohexyl-1,3- dihydro-1 propyl-2H- 1,4-benzodiazepin-2-one (1.49g) was dissolved in hydrobromic acid (4ml of a 30% solution in glacial acetic acid) and stirred at room temperature for 45 min. The yellow solution was then added dropwise to anhydrous diethyl ether (20ml) at OOC and the resultant cream solid filtered off and washed with ether. The solid was partitioned between dichloromethane (50ml) and 10% sodium hydroxide solution (30ml). The organic layer was separated, dried (Na2SO4) and evaporated in vacuo to afford the crude 3(R,S)-amino-5cyclohexyl-1,3-dihydro-1-propyl-2H-1,4-benzodiazepin-2-one (1.06g) as a colourless viscous oil.
A solution of the crude amine (0.7g) in anhydrous tetrahydrofix (13ml) under an atmosphere of nitrogen at room temperature, was treated with triethylamine (323p1), followed by a solution of 4-nitrophenyl chloroformate (0.47g) in anhydrous tetrahydrofuran (13ml). The mixture was stirred for 4h then more triethylamine (32 l) and 4-nitrophenyl chloroformate (47mg) were added. The mixture was stirred for a further 2h then the solid filtered off, washed with tetrahydrofuran, and the filtrate evaporated in vacuo. The residue was azeotroped with toluene (2 x 20ml) then triturated with diethyl ether to afford the title compound (553mg).
Mp 152-155 C. 1H NMR (360MHz, CDCl3) 0.85 (3H, t, J=7.4Hz), 1.02-1.50 (6H, m), 1.56-1.76 (4H, m), 1.84-1.93 (1H, m), 2.00-2.08 (1H, m), 2.76-2.87(IH, m), 3.53-3.63 (1H, m), 4.26-4.36 (1H, m), 5.14 (1H, d, J=8.0Hz), 6.93 (1H, d, J=8.3Hz), 7.22-7.40 (4H, m), 7.50-7.62 (-2H, m); 8.18-8.26(2H, m).
Step 3: 4-(N-tert-Butyloxycarbonyl-N'-piperazinyl)- 1- nitrobenzene To a solution of N-tet-butyloxycarbonylpiperazine (5g) in anhydrous dimethylformamide (75ml) containing triethylamine (7.4ml), was added 1-fluoro-4-nitrobenzene (3.81g). The solution was heated at 1200C for 5h then the solvent was evaporated in vacuo and the residue partitioned between ethyl acetate (50ml) and water (50ml). The organic layer was separated and the aqueous phase washed with ethyl acetate (3 x 50ml). The combined organic layers were washed with water (2 x 50ml), dried (Na2SO4) and evaporated in vacuo. The residue was triturated with ether and the resultant yellow solid filtered off.
The solid was then chromatographed on silica gel, using a gradient elution of 0#1% methanol in dichloromethane to afford the title compound (4.45g) as a yellow solid. 1H NMR (360MHz, CDCl3) 3 1.49 (9H, s), 3.40-3.43 (4H, m), 3.59-3.62 (4H, m), 6.82 (2H, dd, J=9 and 1.8Hz), 8.13 (2H, dd, J=9 and 1.8Hz). MS (Cl, NH3) 307 (M+).
Step 4: 1-Amino-4-(N-tert-butyloxycarbonyl-N'piperazinvl)benzene A suspension of 4-(N-tert-butyloxycarbonyl-N' piperazinyl)-1-nitrobenzene (3.09g) in ethanol (70ml) was hydrogenated at 30psi for 15 min, in the presence of a palladium on carbon catalyst (0.35g, 11% (w/w)). The catalyst was filtered off and the residue evaporated in vacuo. The resultant oil was azeotroped with toluene (2 x 20ml) followed by petrol (60/80) (20ml) to afford the aniline (1.96g). 1H NMR (360MHz, CDCl3) 6 1.48 (9H, sj, 2.96-2.98 (4H, m), 3.56-3.59 (4H, m), 6.65 (2H, d, J=8.7), 6.81 (2H, d, J=8.7Hz). MS (Cl, NH3) 278 (M+1).
Step 5: N-[3(R,S)-5-Cyclohexyl-2,3-dihydro-2-oxo-1-propyl 1H- 1 A-benzodiazepin-3-vllN'-f4-F(N-te rt-butvloxycarbonvl-N '- piperazinvi )phenvllurea To a stirred solution of 5-cyclohexyl-1,3-dihydro-3(R,S)-[(4- nitrophenyloxycarbonyl)amino]-1-propyl-2H-1,4-benzodiazepin-2-one (500mg) in anhydrous dimethylformamide (6ml), under an atmosphere of nitrogen, was added triethylamine (173p1). After 5 min a solution of 1-amino-4-(N-tert-butyloxycarbonyl-N'-piperazinyl)benzene (346mg) in anhydrous dimethylformamide (6ml) was added and the solution heated at 500C for 2h.After this time the solvent was evaporated in vacuo and the residue partitioned between ethyl acetate (20ml) and 20% aqueous acetic acid (20ml). An undissolved white solid (260mg) was collected and identified as the desired urea. 1H NMR (360MHz, CDCl3) 0.81 (3H, t, J=7.4Hz), 1.03-2.04 (21H, m), 2.78 (1H, m), 3.10 (4H, brs), 3.50-3.61 (5H, m), 4.24-4.33 (1H, m), 5.30 (1H, d, J=8Hz), 6.60 (2H, m), 6.92-6.95 (2H, m), 7.23-7.34 (4H, m), 7.49 (1H, dd, J=8.5 and 8.5Hz), 7.55 (1H, dd, J=8 and 1.5Hz). MS (Cl, NH3) 602 (M+).
Step 6: N-[3(R,S)-5-Cyclohexy]-2,3-dihydro-2-oxo-1propyl-1H-1,4-benzodiazepin-3-yl]N'-[4-(N-piperazinyl)phenyl] urea. hvdrochloride salt Into a solution of N-[3(R,S)-5-cyclohexyl-2,3-dihydro-2- oxo-l-propyl-1H-1,4-benzodiazepin-3-yl] N'-t4-(N-tert- butyloxycarbonyl-N'-piperazinyl)phenyl]urea (260mg) in ethyl acetate (40ml) and dichloromethane (5ml), at OOC, was bubbled hydrogen chloride for 40 min. After this time the solvent was evaporated in vacuo and the residue azeotroped with ethyl acetate (3 x 50ml). The resultant white solid was triturated with ether and filtered off.The title compound (156mg) was isolated as a white solid. 1H NMR (360MHz, D6-DMSO) 30.73 (3H, t, J=7.4Hz), 0.90-0.96 (1H, m), 1.13-1.89 (11H, m), 2.95(1H, m), 3.21 (8H, m), 3.65 (1H, m), 4.17-4.21 (1H, m), 5.02 (1H, d, J=8.6Hz), 6.88 (2H, d, J=9Hz), 7.16 (1H, d, J=8.6Hz), 7.23 (2H, d, J=9Hz), 7.37 (1H, m), 7.63 (2H, m), 7.76 (1H, d, J=7.8Hz), 8.84 (1H, s), 8.95 (1H, brs). MS (Cl, NH3) 503 (M+1).
EXAMPLE 5: N-[3(R,S)-5-(cyclohexyl-2,3-dihydro-2-oxo-1propyl-1H-1,4-benzodiazepin-3-yl] N'-[4-(N-methyl-N' pinerazinvl)phenvll urea To a cooled (0 C) and stirred solution of N-[3(R,S)-5 cyclohexyl-2,3-dihydro-2-oxo-1-propyl-1H- 1,4-benzodiazepin-3yl]N['-[4-(N-piperazinyl)phenyl]urea (96mg) in methanol (4ml) and glacial acetic acid (49pl) was added sodium cyanoborohydride (24mg) followed by dropwise addition of a solution of formaldehyde (37p1 of a 38% (w/v) solution in water) in methanol (2ml). The solution was stirred at 0 C for 20 min then at room temperature for 1h. After this time the solvent was removed in vacuo and the residue partitioned between dichloromethane (20ml) and saturated potassium carbonate solution (20ml). The organic phase was separated and the aqueous layer washed with dichloromethane (2 x 20ml). The combined organic layers were washed with brine (20ml), dried (Na2SO4) and evaporated in vacuo to afford a white solid.This was then chromatographed on silica gel, eluting with 90:10:1:1 dichloromethane:methanol: acetic acid:water to afford the title compound (60mg) as a white solid. 1H NMR (360MHz, D6-DMSO) 0.73 (3H, t, J=7.4Hz), 0.89-0.95 (1H, m), 1.10-1.70 (11H, m), 2.20 (3H, 5), 2.40-2.43 (4H, m), 2.94-3.01 (5H, m), 3.60-3.70 (1H, m), 4.15-4.22 (1H, m), 5.02 (1H, d, J=8.9Hz), 6.81 (2H, d, J=9Hz), 7.12 (1H, d, J=8.6Hz), 7.17 (2H, d, J=9Hz), 7.35-7.39 (1H, m), 7.61-7.63 (2H, m), 7.76 (1H, d, J=7.8), 8.74 (1H, s). MS (Cl, NH3) 517 (M+1).
EXAMPLE 6: N-[3(R,S)-5-Cyclohexyl-2,3-dihydro-2-oxo-1-propyl-1H1,4-benzodiazepin-3-yl]N'-[3-N-methyl-N'-piperazinyl)phenyl]urea Step 1: 3-(N-Methyl-N'-piperazinyl)-1-nitrobenzene A mixture of N-methylbis(2-chloroethyl )-amine hydrochloride (9.8g) and 3-nitroaniline (7.0g) in 1-butano1(100ml) was heated at reflux for 60h then cooled to room temperature.
Sodium carbonate (2.8g) was added and the mixture heated at reflux for a further 18h. The mixture was cooled to 0 C and filtered. The solid was collected, washed with anhydrous ether then partitioned between dichloromethane (200ml) and sodium hydroxide solution (1M, 150ml). The organic layer was separated and the aqueous phase washed once more with dichloromethane (200ml). The combined organic layers were dried (K2CO3) and evaporated in vacuo. The resultant residue was chromatographed on silica gel, using dichloromethane:methanol (96:4) as the eluant, to afford an orange oil.The oil crystallized on standing and the resultant solid was triturated with petrol (60/80) to give the desired piperazine (5.64g). 1H NMR (360MHz, CDCl3) 6 2.37 (3H, s), 2.58 (4H, t, J=5Hz), 3.30 (4H, t, J=5Hz), 7.18 (1H, dd, J=9 and 3Hz), 7.35 (1H, t, J=8Hz), 7.64 (1H, dd, J=9 and 2Hz), 7.71 (1H, d, J=2Hz). MS (Cl, NH3) 222 (M+1).
Step 2: 1-Amino-3-(N-methyl-N'-piperazinyl)benzene A solution of 3-(N-methyl-N-piperazinyl)-1-nitrobenzene (1.17g) in ethanol (40ml) was hydrogenated at 25psi for 20 min in the presence of a palladium on carbon catalyst (200mg, 17% (w/w)). The catalyst was filtered off and the solvent evaporated in vacuo. The residue was chromatographed on silica gel, using a gradient elution of petrol:ether (1:1) followed by dichloromethane:methanol (95:5) to give a colourless oil, which was azeotroped with toluene (20ml) then left at 0 C overnight.After this time the oil had crystallized, and after trituration z ion with petrol (60/80j the desired aniline (0.86g) was isolated as a white solid. 1H NMR (360MHz, CDCl3) 82.34 (3H, s), -2.55 (4H, t, J=5Hz), 3.18 (4H, t, J=6Hz), 3.60 (2H, brs), 6.20 (1H, dd, J=8 and 2Hz), 6.25 (1H, t, J=2Hz), 6.36 (1H, dd, J=8 and 2Hz), 7.04 (1H, t, J=8Hz). MS (Cl, NH3) 192 (M+1).
Step 3: N-[3(R,S)-5-Cyclohexyl-2,3-dihydro-2-oxo-1-propyl-1H-1,4benzodiazepin-3-yl]N'-[3-(N-methyl-N'-piperazinyl)phenyl]urea To a solution of 1-amino-3-(N-methyl-N'-piperazinyl) benzene (140mg) in anhydrous tetrahydrofuran (20ml) at 0 C, under an atmosphere of nitrogen, was added triphosgene (71mg).
After 2 min, triethylamine (260p1) was added dropwise and the mixture stirred at 0 C for 5 min then at room temperature for 10 nun. The mixture was then cooled back to 0 C and a solution of crude 3(R,S)-amino-5-cyclohexyl- 1,3-dihydro-l-propyl-2H-1,4- benzodiazepin-2-one (150mg) in anhydrous tetrahydrofuran (5ml) dropwise. The mixture was stirred at OOC for 5 min then at room temperature for 10 min. The mixture was filtered and the filtrate evaporated in vacuo. The residue was partitioned between ethyl acetate (30ml) and water (30ml) and an undissolved solid filtered off.The organic phase from the filtrate was separated and the aqueous phase washed once more with ethyl acetate (101). The combined organic layers were washed with brine (20ml), dried (Na2SO4) and evaporated in vacuo. The residue was chromatographed on silica gel using 95:5:1 dichloromethane:methanol:ammonia as the solvent. After trituration with ether the title urea (77mg) was isolated as a white solid.Mp 255-257"C. 1H NMR (360MHz, D6-DMSO) 8 0.73 (3H, t, J=7.3Hz), 0.92 (1H, m), 1.07-1.90 (11H, m), 2.19 (3H, s), 2.39-2.42 (4H, m), 2.95 (1H, m), 3.02-3.05 (4H, m), 3.62-3.67 (1H, m), 4.16-4.22 (1H, m), 5.02 (1H, d, J=8.6Hz), 6.49 (1H, dd, J=8 and 1.5Hz), 6.01 (1H, d, J=8 and 1.5z), 7.03 (1H, t, J=8Hz), 7.11 (H, m), 7.21 (1H, d, J=8.5Hz), 7.39 (1H, m), 7.62-7.63 (2H, uni), 7.76 (1H, d, J=7.9Hz), 8;82 (1H, s).
EXAMPLE 7: N-[3(R,S)-2,3-Dihydro-1-methyl-5-(4-methylpiperidin1-yl)-2-oxo-1H-1,4-benzodiazepin-3-yl]N'-[3-(N-methyl-N' pinerazinvl)phenvliurea Step 1: Methvl-2-(N-bromoacetvl-N-methvlamino)benzoate A solution of bromoacetyl bromide (209g) in dichloromethane (200ml) was added dropwise to a cooled (ice bath) solution of methyl N-methylanthranilate (168g) in dichloromethane (1400ml). A solution of sodium hydroxide (59g) in water (400ml) was added dropwise to this ice cold solution then after addition the reaction mixture was stirred at room temperature for 20h. The organic phase was separated and washed with 1M hydrochloric acid (500ml), brine (300ml), saturated sodium hydrogen carbonate solution (400ml), dried (Na2SO4) then evaporated to afford the required product as a solid (255g, 92%).
Mp 80-82"C. 1H NMR (360MHz, CDCl3) 6 3.23 (3H, s), 3.54 (1H, d, J=11Hz), 3.60 (1H, d, J=11Hz), 3.90 (3H, s), 7.40 (1H, d, J=8Hz), 7.51 (1H, dd, J=8 and 8Hz), 7.65 (1H, dd, J=8 and 8Hz), 8.04 (1H, d, J=8Hz).
Step 2: 1-Methyl-1,2,3,4-tetrahydro-3H-1,4benzodiazepin-2.5-dione Ammonia gas was bubbled through an ice-cooled solution of methyl 2-(N-bromoacetyl-N-methylamino)benzoate (255g) in methanol (1600ml) until saturated. The cooling bath was removed and the reaction mixture left standing at room temperature for 18h. The precipitate was collected to afford the required product (79g). The filtrate was evaporated and the residue partitioned between dichloromethane (300ml) and 10% citric acid solution (200ml). The organic layer was separated, washed with brine (200ml), dried (Na2SO4) then evaporated to give a solid which was recrystallised from dichloromethane/petroleum ether (60/80) to afford further product (32.5g). Total yield = 111.5g. Mp 190-193 C.
1H NMR (360MHz, CDCl3) 8 3.42 (3H, s), 3.80 (2H, brs), 6.80 (1H, s), 7.24 (1H, d, J=8Hz), 7.32 (1H, dd, J=8 and.8Hz), 7.57 (1H, dd, J=8 and 8Hz), 7.90 (1H, d, J=8Hz).
Step 3: 1.2-Dihvdro-1-methvl-5-(4-methvlpiperidin-1-vl)- 3H-1.4-benzodiazepin-2-one To a solution of 1-methyl-1,2,3,4-tetrahydro-3H- 1,4benzodiazepin-2,5-dione (5g) in anhydrous dichloromethane (150ml) was added phosphorous pentachloride (6.74g) in anhydrous dichloromethane (350ml) over a period of 30 min.
The reaction mixture was stirred at ambient temperature for 1.5h. The solvent was evaporated in vacuo and the residue redissolved in anhydrous dichloromethane (200ml). After cooling to 0 C, a solution of 4-methylpiperidine (10.9ml) in anhydrous dichloromethane (100ml) was added dropwise. When the addition was complete, the reaction mixture was allowed to warm to ambient temperature and left to stir for 2h. The solution was washed with saturated sodium bicarbonate solution (300ml). The organic layer was dried (Na2SO4) and treated with decolourising charcoal (5 spatulas) and silica gel (10 spatulas).
After filtering, the solvent was evaporated in vacuo. The residue was redissolved in dichloromethane (100ml) and extracted with citric acid solution (10%, 4 x 25ml). The combined aqueous layers were basified (4M NaOH solution) and extracted with dichloromethane (6 x 50ml). These organic extracts were combined, dried (K2CO3) and evaporated in vacuo to give the title compound (4.0g) as a beige solid. Mp 106-108 C. 1H NMR (360MHz, CDCl3) 0.97 (3H, d, J=6.3Hz), 1.10-1.22 (1H, m), 1.26-1.42 (1H, m), 1.55-1.63 (2H, m), 1.70-1.77 (1H, m), 2.62-2.70 (1H, m), 2.76-2.86 (1H, m), 3.35 (3H, s), 3.47-3.57 (2H, m), 3.80-3.90 (1H, m), 4.23 (1H, d, J=14.4Hz), 7.18-7.30 (2H, m), 7.45-7.56 (2H, m).
Step 4: 1.2-Dihvdro-1-methvl-5-(4-methvlpiperidin-1-vl)- 3-oximido-3H-1,4-benzodiazepin-2-one A solution of 1,2-dihydro-1-methyl-5-(4-methylpiperidin-1-yl)- 3H-1,4-benzodiazepin-2-one (3.5g) in anhydrous toluene (100ml) was cooled to -200C under an atmosphere of nitrogen, and potassium-tert-butoxide (3.95g) was added portionwise over 2 min.
After stirring at -2O0C for 30 min, isopentyl nitrite (2.1ml) was added dropwise to the red mixture. The reaction mixture was stirred at -20 C for 5h and then allowed to warm to ambient temperature. The mixture was poured onto a rapidly stirred mixture of ethyl acetate (200ml) and water (50ml) containing citric acid (2.45g). This mixture was stirred vigorously for 5 min and then neutralized to pH7 using saturated potassium carbonate solution.
The organic layer was separated and the aqueous layer extracted with ethyl acetate (3 x 100ml). The combined organic phases were dried (Na2SO4) and evaporated in vacuo to give an orange solid.
This was triturated with diethyl ether and the resulting solid collected by filtration to give the title compound (2.39g) as a beige solid. Mp 225-228 C. 1H NMR (360MHz, D6-DMSO) 6 0.90-1.15 (4H, m) 1.20-1.36 (1H, m), 1.54-1.76 (3H, m), 2.77-3.01 (2H, m), 3.26 (3H, s), 3.69-4.24 (2H, m), 7.24-7.32 (1H, m), 7.38-7.43 (1H, m), 7.46-7.58 (2H, m), 9.93 and 10.16 (1H, 2 x 8).
Step 5: 1.2 Dihydro-1-methyl-5-(4-methylpiperidin-1-yl)- 3-O-ethylaminocarbonvl)oximido-3H- 1 .4-benzodiazepin-2-one To a suspension of 1,3-dihydro-1-methyl-5-(4-methylpiperidin- 1-yl)-3-oximido-3H-1,4-benzodiazepin-2-one (0.5g) in anhydrous tetrahydrofuran (20ml) was added triethylamine (0.23ml) followed by ethyl isocyanate (0.2ml) dropwise. This mixture was heated at 600C under an atmosphere of nitrogen for 4.5h. The solvent was evaporated in vacuo to give the title compound (0.62g) as a pale yellow foam. H NMR (360MHz, CDCl3) # 1.00 (3H, brs), 1.13 (3H, t, J=7.2Hz), 1.22-1.42 (2H, m), 1.54-1.90(3H, m), 2.78-3.30 (4H, m), 3.44 (3H, s), 3.56-3.78 (1H, m), 4.54-4.86 (1H, m), 6.37-6.45 (1H, m), 7.21-7.38 (3H, m), 7.46-7.54 (1H, m). TLC (silica, dichloromethane:methanol 90:10) Rf=0.66 and 0.71.
Step 6: N-[3(R,S)-2,3-Dihydro-1-methyl-5-(4-methylpiperidin1-yl)-2-oxo-1H-1,4-benzodiazepin-3-yl] N'-[3-(N-methyl-N' piperazinvlBphenvllurea To a solution of 1,2-dihydro-1-methyl-5-(4. methylpiperidin-1yl)-3-(O-ethylaminocarbonyl)oximido-3H-1,4-benzodiazepin-2-one (0.62g) in methanol (30ml) was added 10% palladium on carbon (0.2g, 32% (w/w)). The mixture was hydrogenated at 40psi for 2h.
Further 10% palladium on carbon (0.1g, 16% (w/w)) was added and the mixture hydrogenated at 40psi for another lh. The catalyst was then filtered off and washed with methanol. The solvent was evaporated in vacuo to give the amine (0.48g) as a pale yellow gum which was used without further purification.
To a solution of 1-amino-3-(N-methyl-N'-piperazinyl)benzene (Example 2, Step 2) (0.48g) in anhydrous tetrahydrofuran (30ml) cooled to one under an atmosphere of nitrogen was added triphosgene (0.25g) in one portion. Triethylamine (1.0ml) was added dropwise to ensure a basic solution. After stirring at 0 C for 30 min a solution of the amine (0.48g) in anhydrous tetrahydrofuran (10ml) was added dropwise. The mixture was stirred at 0 C for 5 min and then allowed to warm to ambient temperature and stirred for 10 min. The solvent was evaporated in vacuo and the residue partitioned between ethyl acetate (50ml) and water (50ml).The undissolved solid was collected by filtration and purified by chromatographing on silica gel using 95:5:1, dichloromethane:methanol:aqueous ammonia solution increasing the ratio to 90:10:1, to give the title compound (0.55g) as a colourless solid.
Mp 16000 (dec.). 1H NMR (360MHz, CDCl3) 80.95 (3H, d, J=6.2Hz), 1.05-1.21 (1H, m), 1.28-1.37 (1H, m), 1.46-1.62 (2H, m), 1.64-1.74 (1H, m), 2.45 (3H, s), 2.57-2.80 (6H, m), 3.25-3.35 (4H, m), 3.41 (3H, s), 3.47-3.60 (1H, m), 3.86-3;96-(1H, m), 5.26 (H, d, J=7.8Hz), 6.52-6.58 (2H, m), 6.65 (1H, d, J=7.9Hz), 6.98 (1H, brs), 7.11 (1H, t, J=8.1Hz), 7.16-7.20 (1H, m), 7.21-7.33. (2H, m), 7.47-755 (2H, m) EXAMPLE 8:N-[3(R,S)-5-Cycloheptyl-2,3-dihydro-1-methyl-2-oxo lH-1.4-benzodiazepin-3-vllN'-[3-(N-morpholinomethvl)phenvll urea Step 1: 3(R,S)-[(Benzyloxycarbonyl)amino]-5-cyclohepty]1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one 3(R,S)-[(Benzyloxycarbonyl)amino]-5-cycloheptyl-1,3dihydro-2H-1,4-benzodiazepin-2-one (Example 1, Step 3, 500mg) in anhydrous toluene (40ml) was heated to reflux. A solution of dimethylformamide dimethyl acetal (786 l) in anhydrous toluene (lOml) was added dropwise and the mixture was heated at reflux for a further hour.
The solvent was evaporated and the residue triturated with diethyl ether to afford the title compound (441mg) as a colourless solid. 1H NMR (360MHz, CDCl3) 3 1.24-1.90 (11H, m), 2.00-2.14 (1H, m), 2.90-3.00 (1H, m), 3.40 (3H, s), 5.04-5.18 (3H, m), 6.52 (1H, d, J=7.5Hz), 7.24-7.60 (9H, m). TLC (silica, petrol (60/80): ethyl acetate 2:1). Rf = 0.30.
Step 2: 3-(N-Mornholinomethvl- l-nitrobenzene A solution of morpholine (2.0g) in anhydrous acetone (100ml) was treated with potassium carbonate (4.8g) and 3-nitrobenzyl bromide (5.0g). The mixture was stirred at room temperature for 48h. The solvent was evaporated and the residue partitioned between ethyl acetate (100ml) and water (50ml). The aqueous layer was washed with ethyl acetate (2 x 80ml). The combined organics were dried (Na2SO4), evaporated and the residue chromatographed on silica gel with 3:1 petrol:ethyl acetate as eluant to afford the title compound (2.6g) as a pale yellow solid. 1H NMR (250MHz, CDCl3) 8 2.42-2.54 (4H,m), 3.61 (2H, s), 3.70-3.80 (4H, m), 7.50 (1H, t, J=7.5Hz), 7.69 (1H, d, J=7.5Hz), 8.09-8.18 (1H, m), 8.24 (1H, s). TLC (silica, petrol (60/80):ethyl acetate 3:1). Rf= 0.50.
Step 3: 1-Amino-3-(N-morpholinomethyl)benzene 3-(N-Morpholinomethyl)-1-nitrobenzene (1.25g) in ethanol (50ml) was treated with 10% palladium on carbon (125mg, 10% (w/w)). The mixture was hydrogenated at 25psi for 7 min. The catalyst was filtered off and the solvent evaporated. The residue was chromatographed on silica gel with 2:1 petrol:ethyl acetate as eluant to afford the title compound (0.75g). H NMR (360MHz, CDCl3) # 2.40-2.50 (4H, m), 3.42 (2H, s), 3.64 (2H, brs), 3.68-3.80 (4H, m), 6.57 (1H, dd, J=7.2 and 1.6Hz), 6.69-6.71 (2H, m), 7.07-7.11 (1H, m). TLC (silica, petrol (60/80):ethyl acetate 2:1). Rf=0.30.
Step 4: N-[3(R,S)-5-Cycloheptyl-2,3-dihydro-1-methyl-2-oxo1H-1,4-benzodiazepin-3-yl]N'-[3-(N-morpholinomethyl)phenyl]urea 3(R,S)-[(Benzyloxycarbonyl)amino]-5-cycloheptyl-1,3-dihydro1-methyl-2H-1,4-benzodiazepin-2-one (435mg) was treated with a solution of 30% hydrogen bromide in glacial acetic acid (lOml) and stirred for 20 min at room temperature. The mixture was then added dropwise onto cold (0 C) diethyl ether (50ml). A white solid precipitated which was filtered off. The solid was treated with 10% sodium hydroxide solution (50ml), then extracted with ethyl acetate (80ml). The organic layer was separated, dried (Na2SO4) and evaporated to give crude 3(R,S)-amino-5-cycloheptyl-1,3-dihydro-1- methyl-2H-1,4-benzodiazepin-2-one as a colourless oil.
l-Amino-3-(N-morpholinomethyl)benzene (180mg) in anhydrous tetrahydrofuran (40ml) was cooled to OOC whereupon triphosgene (92mg) was added. The mixture was stirred at OOC for 2 min, then triethylamine (391p1) was added portionwise until pH8. The mixture was then stirred for a further 5 min, allowed to warm to 15 C, and then re-cooled to 000. Then a solution of 3(R,S)-amino-5-cycloheptyl-1,3-dihydro-1-methyl-2H- 1,4- benzodiazepin-2-one (181mg) in anhydrous tetrahydrofuran (7ml) was added dropwise over 5 min. The mixture was stirred at 0 C for 5 min, allowed to warm to room temperature and then stirred for a further 2h. The undissolved material was removed by filtration. The solvent was evaporated in vacuo and the residue partitioned between ethyl acetate (50ml) and water (20ml). The organic phase was separated, dried (Na2SO4) and evaporated. The crude solid was recrystallised from ethyl acetate to afford the title compound as a colourless solid (89mg).
Mp 190-192 C. 1H NMR (360MHz, CDCl3)#1.24-1.94 (llH, m), 2.04-2.16 (1H, m), 2.40-2.48 (4H, m), 2.90-3.00 (1H, m), 3.42 (3H, s), 3.45 (2H, s), 3.66-3.73 (4H, m), 5.35 (1H, d, J=8Hz), 6.61 (1H, d, J=8Hz), 6.81 (1H, s), 6.99-7.06 (1H, m), 7.18-7.34 (4H, m), 7.36 (1H, s), 7.46-7.62 (2H, m).
EXAMPLE 9: N-(R,S)-5-Cycloheptyl-2,3-dihydro-1-methyl-2-oxo-1H1,4-benzodiazepin-3-yl]N'-[3-(2-(N-morpholine)ethoxy)phenyl]urea Step 1: 3-[2-(N-Morpholino)ethoxy]-1-nitrobenzene A solution of 3-nitrophenol (3.0g) in anhydrous dioxane (lOOm') was treated with caesium carbonate (18g) and N-(2chloroethyl)morpholine (20g). The mixture was heated at 80 C for 16h. The undissolved material was filtered off. The solvent was evaporated in vacuo and the residue partitioned between ethyl acetate (80ml) and water (50ml). The organic phase was separated, dried (Na2SO4) and evaporated.The residue was chromatographed on silica gel with petrol (60/80):ethyl acetate (1:1) as the eluant to afford the title compound (3.2g) as a yellow oil. 1H NMR (250MHz, CDCl3) 6 2.55-2.65 (4H? m), 2.84 (2H, t, J=5Hz), 3.72-3.78 (4H, m), 4.20 (2H, t, J=5Hz), 7.21-7.29 (1H, m), 7.42 (1H, t, J=7.5Hz), 7.72-7.78 (1H, m), 7.80-7.88 (1H, m). TLC (silica, ethyl acetate) Rf = 0.36.
Step 2: 1-Amino-3-[2-(N-morpholino)ethoxy]benzene 3-[2-(N-Morpholino)ethoxy]-1-nitrobenzene (1.6g) in ethanol (30ml) was treated with 10% palladium on carbon (159mg, 10% (w/w)). The mixture was hydrogenated at 25psi for 15min. The catalyst was filtered off and the solvent evaporated. The residue was chromatographed on silica gel with ethyl acetate as the eluant to afford the title compound (1.05g). 1H NMR (250MHz, CDCl3) 3 2.54-2.63 (4H, m), 2.80 (2H, t, J=6.3Hz), 3.65 (2H, brs), 3.72-3.78 (4H, m), 4.09 (2H, t, J=6.3Hz), 6.21-6.36 (3H, m), 7.05 (1H, t, J=7.5Hz). TLC (silica, ethyl acetate) Rf = 0.18.
Step 3: N-r3(R.S)-5-Cveloheptvl-2.3-dihvdro-1-methvl-2-oxo- 1H-1,4-benzodiazepin-3-yl]N[-[3-(2-(N-morpholino)ethoxy)phenyl]urea l-Amino-3-[2-(N-morpholino)ethoxy]benzene (323mg) in anhydrous tetrahydrofuran (30ml) was cooled to OOC whereupon triphosgene (143mg) was added. The mixture was stirred at OOC for 2 min, then triethylamine (607 l) was added portionwise until pH8. The mixture was then stirred for a further 5 min, allowed to warm to 15 C, and then re-cooled to 0 C. Then a solution of 3(R,S)-amino-5-cycloheptyl-1,3-dihydro-1-methyl-2H- 1,4-benzodiazepin-2-one (282mg) in anhydrous tetrahydrofuran (10ml) was added dropwise over 5 min.The mixture was stirred at OOC for 5 min, allowed to warm to room temperature and stirred for a further lh. The undissolved material was removed by filtration. The solvent was evaporated in vacuo and the residue partitioned between ethyl acetate (50ml) and water (20ml). The organic phase as separated, dried (Na2S94) and evaporated. The residue was chromatographed on silica gel with a gradient elution of 0#3% methanol in dichloromethane to afford the title compound (360mg).Mp 204-205 C. 1H NMR (-360MHz, D6-DMSO) # 1.10-1.86 (llH, m), 1.90-2.02 (1H, m), 2.40-2.48 (4H, m), 2.65 (2H, t, J=5.7Hz), 3.08-3.20 (1H, m), 3.30 (3H, s), 3.54-3.60 (4H, m), 3.99 (2H, t, J=5.7Hz), 5.04 (1H, d, J=8.4Hz), 6.48 (1H, dd, J=7.7Hz and 2.0Hz), 6.75 (1H, d, J=9.4Hz), 7.06-7.15 (2H, m), 7.26 (1H, d, J=8.4Hz), 7.37 (1H, t, J=6.7Hz), 7.54 (1H, d, J=8.3Hz), 7.63 (1H, t, J=7.0Hz), 7.76 (1H, dd, J=7.9Hz and 1.4Hz), 8.98 (1H, s).
EXAMPLE 10: N-[3(R)-5-Cyclohexyl-2,3-dihydro-1-methyl-2-oxo-1H1,4-benzodiazepin-3-yl]N'-[3-(2-(N-morpholino)ethoxy)phenyl]urea Step 1: 3(R,S)-Amino-5-cyclohexyl-1,3-dihydro-1-methyl 2H-l .4-benzodiazepin-2-one A mixture of 3(R,S)-[(benzyloxycarbonyl)amino]-5- cyclohexyl-1,3-dihydro- l-methyl-2H-1,4-benzodiazepin-2-one (3.0g) and hydrobromic acid (45% in acetic acid, 6.2ml) was stirred for 1H at room temperature under an atmosphere of nitrogen. The mixture was then diluted with cold anhydrous diethyl ether (40ml) and stirred at OOC for 45 min.The white precipitate was collected by filtration, washed with cold diethyl ether (4 x 30ml) and then dissolved in a mixture of water (30ml) and aq. sodium hydroxide (2M, 15ml). The basic aqueous phase was extracted with ethyl acetate (3 x 70ml) and the combined organic layers were washed with brine (30ml), dried (Na2SO4) and concentrated. The residue was chromatographed on silica gel, using 94:6, dichloromethane:methanol as the eluant, to afford the title compound (1.6g) as a pale pink solid.
Mp 133-1360C. 1H NMR (360MHz, CDCl3) 61.02-1.40 (4H, m), 1.47-1.56 (1H, m), 1.61-1.74 (3H, m), 1.84-1.91 (1H, m), 1.96-2.06 (1H, m), 2.17 (2H, brs), 2.70-2.80 (1H, m), 3.39 (3H, s), 4.29 (1H, s), 7.20-7.27 (2H, m), 7.44-7.54 (2H, m).
Step 2: 3(R,S)-[2(R)-(tert-Butyloxycarbonyl)amino-3phenylpropionylamino]-5-cyclohexyl-1,3-dihydro-1-methyl-2H 1 .4-benzodiazepin-2-one To a solution of 3(R,S)-amino-5-cyclohexyl-1,3-dihydro-1 methyl-2H-1,4-benzodiazepin-2-one (4g) in anhydrous dimethylformamide (35ml), under an atmosphere of nitrogen, was added in succession Boc-D-phenyl-alanine (4.11g), 1-hydroxybenzotriazole trihydrate (2.09g) and 1-ethyl-3-[3 (dimethylamino)propy13carbodiimide hydrochloride (2.97g).
Triethylamine (2.16ml) was then added and the resulting suspension was stirred at ambient temperature for 20 min. The solvent was removed under reduced pressure and the residue was partitioned between ethyl acetate (50ml) and 10% citric acid solution (50ml). The organic phase was separated and the aqueous phase extracted with ethyl acetate (3 x 50ml). The combined organic phases were washed with 10% sodium hydroxide solution (50ml), water (50ml) and brine (50ml), dried (MgSO4) and evaporated in vacuo.The residue was chromatographed on silica gel, using 1:1 petrol:ethyl acetate as the eluant, to afford the product (7.26g), mp 95-98 C. 1H NMR (360MHz, CDCl3) #0.99-1.11 (1H, m), 1.16-1.72 (7H, m), 1.40 (9H, s), 1.83-1.92 (1H, m), 1.98-2.06 (1H, m), 2.73-2.83 (1H, m), 3.10-3.24) (2H, m), 3.38 (3H, s), 4.53 (1H, brs), 4.98 (1H, brs), 5.28-5.34 (2H, m), 7.19-7.32 (7H, m), 7.49-7.58 (2H, m).
Step 3: (+)-3(R)-(2(R)-Amino-3-phenylpropionylamino)-5cyclohexyl-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one 3(R,S)-[2(R)-(tert-Butyloxycarbonyl)amino-3 phenylpropionylsmino]-5-cyclohexyl-1,3-dihydro-1-methyl-2H- 1,4-benzodiazepin-2-one (4.7g) was dissolved in ethyl acetate (20ml) and cooled to OOC. This solution was then saturated with hydrogen chloride gas. After 1.5h, the resulting precipitate (which was shown to be the undesired diastereoisomer, Rf = 0.04 ethyl acetate), was removed by filtration and the filtrate evaporated. The solid residue was partitioned between ethyl- acetate (25ml) and 10% sodium carbonate solution (20ml). The organic phase was separated and the aqueous extracted with ethyl acetate (2 x 25ml).
The combined organic phases were dried (Na2SO4) and evaporated in vacuo. The residue was chromatographed on silica gel using a gradient elution of 0-20% methanol in ethyl acetate to afford the title compound (1.66g). Mp 100-103 C. 1H NMR (360MHz, CDCl3) 3 1.00-1.39 (4H, m), 1.50-1.72 (4H, m), 1.84-1.92 (1H, m), 2.00-2.07 (1H, m), 2.72-2.84 (2H, m), 3.28 (1H, dd, J=13.8 and 4.0Hz), 3.40 (3H, s), 3.69 (1H, dd, J=9.8 and 4.1Hz), 5.36 (1H, d, J=8.3Hz), 7.21-7.36 (7H, m), 7.47-7.58 (2H, m), 8.66 (1H, d, J=8.3Hz).
[a]D23 +32.7 (c = 0.58, CH30H).
Step 4: (+)-N-[1(R)-2[(3(R)-5-Cyclohexyl-2,3-dihydro-1methyl-2-oxo-1H-1,4-benzodiazepin-3-yl)amino]-2-oxo-1 (phenylmethyl)ethyl]N'-phenyl thiourea A solution of (+)-3(R)-(2(R)-amino-3- phenylpropionylamino)5-cyclohexyl-1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one (1.6g) in anhydrous dichloromethane (lOml) was treated with phenyl isothiocyanate (0.5ml), and then heated on the steam bath for 30 min. The solvent was evaporated in vacuo and the residue was chromatographed on silica gel with 1:1, ethyl acetate:petrol as the eluant, to afford the product (2.1g) as a pale yellow solid.
Mp 129-132 C. 1H NMR (360MHz, CDCl3) # 0.95-1.07 (1H, m), 1.15-1.37 (3H, m), 1.45-1.69 (4H, m), 1.81-1.88 (1H, m), 1.93-2.00 (lH, m), 2.70-2.80 (1H, m), 3.24-3.41 (2H, m), 3.38 (3H, s), 5.23 (1H, d, J=7.3Hz), 5.31-5.40 (1H, m), 6.67 (1H, 7.0Hz), 6.87-7.02 (2H, m), 7.20-7.35 (9H, m), 7.46-7.52 (2H, m), 7.65 (1H, s).
[&alpha;]25D+27.3 (c = 0.31, CH2Cl2).
Step 5: (+)-3(R)-Amino-5-cyclohexyl-1,3-dihydro-1-methyl 2H-1A-benzodiazepin-2-one (+)-N-[1(R)-2-[(3(R)-5-Cyclohexyl-2,3-dihydro-1-methyl-2oxo-1H-1,4-benzodiazepin-3-yl)amino]-2-oxo-1-(phenylmethyl) ethyl]N'-phenyl thiourea (4.5g) was dissolved in trifluoroacetic acid (25ml) and stirred at ambient temperature for 30 min. The trifluoroacetic acid was removed under reduced pressure and the residue azeotroped with dichloromethane (2 x 20ml) and toluene (2 x 20ml). The residue was chromatographed on silica gel using 90:10:0.1:0.1, dichloromethane:methanol: acetic acid:water as the eluant, to afford an orange gum. This was dissolved in ethyl acetate (150ml), cooled to 0 C, and treated with 10% sodium carbonate solution (15ml).After diluting with water (25ml) and stirring for 1 min, the organic layer was separated and the aqueous re-extracted with ethyl acetate (2 x 50ml). The combined organics were dried (Na2SO4) and evaporated in vacuo to afford the title compound (1.56g) as a solid with 99% e.e. Mp 133-136 C. 1H NMR (360MHz, CDCl3) # 1.01-1.39 (4H, m), 1.50-1.54 (1H, m), 1.60-1.70 (3H, m), 1.84-1.92 (1H, m), 1.96-2.04 (1H, m), 2.36 (2H, brs), 2.70-2.80 (1H, m), 3.41 (3H, s), 4.32 (1H, s), 7.22-7.28 (2H, m), 7.46-7.58 (2H, m). [a]D +83.2 C (c = 0.66, CH3OH).
Step 6: N-[3(R)-5-Cyclohexyl-2,3-dihydro-1-methyl-2-oxo1H-1,4-benzodiazepin-3-yl]N'-[3-(2-(N-morpholino)ethoxy)phenyl]urea 1-Amino-3-[2-(N-morpholino)ethoxy)]benzene (241mg) was dissolved in anhydrous tetrahydrofuran (40ml) and cooled to 0 C under an atmosphere of nitrogen. Triphosgene (106mg) was added in one portion and the mixture was stirred for 5 min. The mixture was then treated with triethylamine (0.45ml) dropwise over a period of5 min. The mixture was allowed to warm to 1000 over a period of 15 min and was then re-cooled to 0 C. 3(R)-Amino-5-cyclohexyl-1,3dihydro-1-methyl-2H-1,4-benzodiazepin-2-one (0.2g) was dissolved in anhydrous tetrahydrofuran (10ml) and added to the reaction dropwise.
The mixture was stirred at ambient temperature for 15 min. The precipitated solid was removed by filtration and washed with tetrahydrofuran. The filtrate was evaporated and partitioned between ethyl acetate (5Oml) and water (25ml). The organic layer was separated and then dried (Na2SO4).-The solvent was evaporated in vacuo and the residue chromatographed on silica gel using a gradient elution of l-+10% methanol in dichloromethane.
The desired urea (220mg) was isolated as a white solid.
Mp 174-176 C. lH NMR (360MHz, D6-DMSO) #0.82-0.93 (1H, m), 1.09-1.89 (9H, m), 2.44 (4H, m), 2.64 (2H, t, J=5.7Hz), 2.90-2.96 (1H, m), 3.29 (3H, m), 3.55 (4H, m), 3.99 (2H, t, J=5.7Hz), 5.05 (1H, d, J=8.3Hz), 6.48 (1H, dd, J=8.0 and 2.1Hz), 6.75 (1H, m), 7.11 (2H, m), 7.26 (1H, d, J=8.4Hz), 7.38 (1H, dd, J=7.1 and 7.1Hz), 7.54 (1H, m), 7.62 (1H, dd, J=7.1 and 7.1Hz), 7.75 (1H, m), 8.98 (1H, s).
EXAMPLE 11: N-[3(R,S)-5-Cycloheptyl-2,3-dihydro-1 (2-methylpropyl)-2-oxo-1H-1,4-benzodiazepin-3-yl]N'-[3-(Nmethyl-N'-piperazinyl)phenyl]urea Step 1: 3(R,S)-[(Benzyloxycarbonyl)amino]-5-cycloheptyl 1.3-dihvdro- 1-(2-methvlpropvl)-2H-1.4-benzodiazepin-2-one To a solution of 3(R,S)-[(benzyloxycarbonyl)amino]-5- cycloheptyl-1,3-dihydro-2H-1,4-benzodiazepin-2-one (1.26g) at OOC, under an atmosphere of nitrogen, was added sodium hydride (125mg of a 50% dispersion in mineral oil). After stirring for 1h at OOC 2-methylpropyl iodide (399p1) was added and the mixture allowed to warm to ambient temperature over 3h.More sodium hydride (16mg of a 50% dispersion in mineral oil) followed by 2-methylpropyl iodide (40p1) was added and the solution stirred at room temperature for 18h. More sodium hydride (103mg of a 50% dispersion in mineral oil) followed by 2-methylpropyl iodide (210 l) was added and the solution stirred for 4h at room temperature. The solvent was then evaporated in vacuo and the residue partitioned between dichloromethane (30ml) and water (30ml). The organic phase was separated and the aqueous layer washed further with dichloromethane (3 x 20ml). The combined organic layers were washed with brine (20ml), dried (MgSO4) and evaporated under reduced pressure.The residue was azeotroped with toluene (2 x 20ml), then chromatographed on silica gel using 4:1 petrol:ethyl acetate as the eluant. The product was isolated as a viscous oil which solidified on addition of petrol:ethyl acetate (4:1) (20ml). After evaporation of the solvent the resultant solid was triturated with petrol (60/80) and the title compound (830mg) isolated as a white solid. 1H NMR (360MHz, CDCl3) #0.73 (3H, d, J=6.6Hz), 0.79 (3H, d, J=6.6Hz), 1.43-1.83 (12H, m), 2.06-2.20 (1H, m), 2.94-3.06 (1H, m), 3.43 (1H, dd, J=13.8 and 5Hz), 4.27 (1H, dd, J=13.8 and 9.3Hz), 5.10 (3H, m), 6.56 (1H, d, J=8.2Hz), 7.24=7.35 (7H, m), 7.49 (1H, dd, J=8.4 and 8.4Hz), 7.58 (1H, d, J=7.9Hz). MS (Cl, NH3) 462 (M+).
Step 2: 5-Cycloheptyl-1,3-dihydro-1-(2-methylpropyl)-3-(R,S) [(4-nitrophenyloxycarbonyl)amino]-2H-1,4-benzodiazepin-2-one 3(R,S)-pSenzyloxycarbonyl)amino]-5-cycloheptyl-1,3- dihydro- 1-(2-methylpropyl)-2H- 1,4-benzodiazepin-2-one (830mg) was dissolved in hydrobromic acid (4ml of a 30% solution in glacial acetic acid) and stirred at room temperature for 20 min.
The yellow solution was then added dropwise to anhydrous diethyl ether (50ml) at OOC and the resultant cream solid filtered off and washed with ether. The solid was partitioned between dichloromethane (50ml) and 10% sodium hydroxide solution (50ml). The organic layer was separated, dried (Na2SO4) and evaporated in vacuo to afford crude 3(R,S)-amino-5-cycloheptyl1,3-dihydro-1-(2-methylpropyl)-2H-1,4-benzodiazepin-2-one as a viscous oil.
The amine was dissolved in anhydrous tetrahydrofuran (9ml) under an atmosphere of nitrogen, at room temperature, and triethylamine (249pl) was added. dropwise. A epiution of 4-nitrophenyl chloroformate (363mg) in anhydrous tetrahydrofuran (9ml) was then added, and the reaction mixture stirred for a further 2h. After this time the undissolved solid was filtered off, washed with tetrahydrofuran, and the filtrate evaporated in vacuo.
The residue was azeotroped with toluene (2 x 20ml) and ether (20ml). The resultant solid was then triturated in anhydrous ether to afford the title compound (420mg) as a white solid. TLC (silica, petrol (60/80):ethyl acetate 2:1) Rf = 0.7. 1H NMR (360MHz, CDCl3) 8 0.77 (3H, d, J=6.6Hz), 0.83 (3H, d, J=6.6Hz), 1.40-1.87 (12H, m), 2.06-2.17 (1H, m), 2.98-3.02 (1H, m), 3.48 (1H, dd, J=13.8 and 5.0Hz), 4.32 (1H, dd, J=13.8 and 9.3Hz), 5.10 (1H, d, J=8.4Hz), 6.92 (1H, d, J=8.2Hz), 7.26-7.39 (4H, m), 7.53 (1H, dd, J=8.5 and 8.5Hz), 7.61 (1H, m), 8.21 (2H, d, J=9Hz).
Step 3: N-[3(R,S)-5-Cycloheptyl-2,3-dihydro-1-(2methylpropyl)-2-oxo-1H-1,4-benzodiazepin-3-yl]N'-[3-(N-methyl N'-pierazinvi)nhenvliurea To a stirred solution of 5-cycloheptyl-l ,3-dihydro-l-(2- methylpropyl)-3(R,S)-[(4-nitrophenyloxycarbonyl)amino]-2H-1,4benzodiazepin-2-one (150mg) in anhydrous dimethylformamide (3ml), under an atmosphere of nitrogen, was added triethylamine (42p1). After 5 min a solution of l-amino-3-(N-methyl-N'piperazinyl)benzene (64mg) in anhydrous dimethylformamide (2ml) was added and the solution heated at 50 C for Th. The solvent was evaporated in vacuo and the residue partitioned between ethyl acetate (20ml) and water (20ml).The undissolved solid was filtered off, washed with ethyl acetate and water, then triturated with anhydrous diethyl ether to yield the desired urea (87mg) as a white solid. 1H NMR (360MHz, D6-DMSO) #0.66 (3H, d, J=6.6Hz), 0.77(3H, d, J=6.6Hz), 1::20-1;80 (12H, m), 1.96-2.10 (1H, m), 2.20 (3H, s), 2.18-2.20 (4H, m), 3.03-3.05 (4H, m), 3.14-3.18 (1H, m), 3.62 (1H, dd, J=13.9 and 4.4Hz), 4.13 (lH,-dd, J=13.9 and 9.6Hz), 5.00 (1H, d, J=8.5Hz), 6.49 (1H, d, J=8.3Hz), 6.61 (1H, d, J=7.7Hz), 7.03 (1H, t, J=8.1Hz), 7.11 (1H, m), 7.23 (1H, d, J=7.7Hz), 7.37 (1H, dd, J=7 and 7Hz), 7.59-7.68 (2H, m), 7.78 (1H, d, J=6.8Hz), 8.84 (1H, s).
MS (FAB) 545 (M+).
EXAMPLE 12: (+)-N-[5-Cyclohentyl-2,3-dihydro-1-methyl-2oxo-1H-1,4-benzodiazepin-3-yl] N'-[3-(N-methyl-N' piperazinvl)phenvllurea Step 1: (+)-2-(2((R)-Amino-3-phenylpropionylamino]-5 cyclohentvl-1 .3-dihydro- 1-methvl-2H- 1 A-benzo diazepin-2-one To a solution of 3(R,S)-amino-5-cycloheptyl-1,3-dihydro-1- methyl-2H-1,4-benzodiazepin-2-one (4.08g) in anhydrous dimethyl formamide (30ml), under an atmosphere of nitrogen, was added in succession Boc-D-phenylalanine (3.98g), 1-hydroxybenzotriazole trihydrate (2.03g) and 1-ethyl-3-[3-(dimethylsmino)propyl] carbodiimide hydrochloride (2.88g). Triethylamine (2.09ml) was then added and the resulting suspension stirred at ambient temperature for lh.The solvent was removed under reduced pressure and the residue partitioned between ethyl acetate (100ml) and 10% citric acid solution (100ml). The organic phase was separated and the aqueous phase extracted with ethyl acetate (3 x 50ml). The combined organic phases were washed with 10% sodium hydroxide solution (100ml), water (50ml) and brine (50ml). The organic layer was separated, dried (Na2SO4) and evaporated in vacuo to afford crude 3(R,S)-[2(R) (tert-butyloxycarbonyl)amino-3-phenylpropionylamino]-5-cycloheptyl1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one 3(R,S)-[2((R)-(tert-Butyloxycarbonyl)amino-3 phenylpropionylamino]-5-cycloheptyl-1,3-dihydro-1-methyl-2H- 1,4-benzodiazepin-2-one (8.6g) was dissolved in ethyl acetate (40ml) any cooled to 000. This solution was then saturated with hydrogen chloride gas. After 1h the resulting precipitate was removed by filtration (which was shown to be the undesired diastereoisomer (Rf = 0.13, ethyl acetate)) and the filtrate evaporated.The solid residue was partitioned between ethyl acetate (25ml) and 10% sodium carbonate solution (20ml). The organic phase was separated and the aqueous layer extracted with ethyl acetate (2 x 25ml). The combined organic phases were dried (Na2SO4) and evaporated in vacuo.The residue was chromatographed on silica gel using a gradient elution of 0#20% methanol in ethyl acetate to afford the title compound (2.5g) as a yellow oil. 1H NMR (250MHz, CDCl3) 6 1.05-1.40 (4H, m), 1.52-1.70 (6H, m), 1.82-1.92 (1H, m), 2.02-2.10 (1H, m), 2.80-2.86 (1H, m), 2.90-2.95 (1H, m), 3.20-3.40 (4H, m), 4.20 (1H, m), 5.30 (1H, d, J=8Hz), 7.20-7.40 (7H, m), 7.41-7.60 (2H, m), 8.00-8.10 (1H, m). TLC (silica, ethyl acetate) Rf = 0.22. [a]23D - +28.0 C (c = 0.2, CH3OH).
Step 2: 3-Amino-5-cycloheptyl-1,3-dihydro-1-methyl-2H 1 .4-benzodiazepin-2-one (Enantiomer A) A solution of (+)-3-[2(R)-amino-3-phenylpropionylamino cycloheptyl- 1,3-dihydro- 1-methyl-2H- l,4-benzodiazepin-2.one (2.5g) in anhydrous dichloromethane (10ml) was treated with phenyl isothiocyanate (0.76ml), and then heated on the steam bath for 30 min. The solvent was evaporated in vacuo and the residue chromatographed on silica gel with 1:1 petrol (60/80):ethyl acetate as eluant to afford N-(1(R)-2-((5-cycloheptyl- 2,3-dihydro-1-methyl-2-oxo-1H-1,4-benzodiazepin-3-yl)amino]-2oxo-1-(phenylmethyl)ethyl]N'-phenyl thiourea (2.3g).
N-[1(R)-2-[(5-Cycloeptyl-2,3-dihydro-1-methyl-2-oxo-1H1,4-benzodiazepin-3-yl)amino]-2-oxo-1-(phenylmethyl)ethyl]N'phenyl thiourea (2.3g) was dissolved in trifluoracetic acid (10ml), and stirred at room temperature for 30 min, then the solvent was removed under reduced pressure and the residue azeotroped with dichloromethane (2 x 20ml) and toluene (2 x 20ml). The residue was chromatographed on silica gel using 90:10:1.0:1.0 - dichloromethane:methanol:acetic acid:water as the eluant to afford an orange gum. This was dissolyed in ethylacetatw(80ml), cooled to OOC, and treated with 10% sodium carbonate solution (8ml).
After diluting with water (25ml) and stirring for 1 min, the organic layer was separated and the aqueous phase re-extracted with ethyl acetate (2 x 50ml). The combined organics were dried (Na2SO4) and evaporated in vacuo to afford the title compound (1.Og) as a yellow oil. 1H NMR (250MHz, CDCl3) # 1.20-1.70 (10H, m), 1.71-1.94 (1H, m), 1.95-2.13 (1H, m), 2.86-3.01 (1H, m), 3.40 (3H, s), 4.36 (1H, brs), 7.13-7.32 (2H, m), 7.41-7.57 (2H, m). TLC (silica, dichloromethyane:methanol 9:1) Rf=0.50.
Step 3: (+)-N-r5-Cveloheptvl-2.3-dihvdro-1-methvl-2-oxo-1H- 1.4-benzodiazepin-3-yl]N'[3-(N-methyl-N'-piperazinyl)phenyl]urea 1-Amino-3-(N-methyl-N -piperazinyl)benzene (197mg) in anhydrous tetrahydrofuran (30ml) was cooled to OOC whereupon triphosgene (101mg) was added. The mixture was stirred at OOC for 2 min, then triethylamine (430 l) was added portionwise until pH8. The mixture was then stirred for a further 5 min, allowed to warm to 15 C, and then re-cooled to 0 C. Then a solution of 3-amino-5-cycloheptyl-1,3-dihydro-1-methyl-2H-1,4benzodiazepin-2-one (200mg) (Enantiomer A) in anhydrous tetrahydrofuran (10ml) was added dropwise over 5 min.The mixture was stirred at OOC for 5 min, allowed to warm to room temperature and then stirred for a further 15 min. The undissolved material was removed by filtration. The filtrate was evaporated in vacuo and the residue partitioned between ethyl acetate (50ml) and water (30ml). The organic phase was separated, dried (Na2SO4) and evaporated. The residue was chromatographed on silica gel with dichloromethane:methanol (9:1) as eluant to afford the title compound (239mg) as a colourless solid.Mp 160 C (dec.). 1H NMR (250MHz, D6-DMSO) 3 1.05-1.87 (11H, m), 1.88-2.04 (1H, m) 2.21 (3H, s), 2.36-2.4i (4H, m), 2.95-3.21 (5H, m), 3.36 (3H, s), 5.05 (1H, d, J=8Hz), 6.50 (1H, dd, J=8.2 and 2.0Hz), 6.61 (1H, d,J=8Hz), 7.04.(lH,.t,.J=7.SHz), 7.13 (1H, s), 7.25 (1H, d, J=7.5Hz), 7.39 (1H, t, J=7.0Hz), 7.52-7.70 (2H, m), 7.76 (1H, d, J=8.0Hz), 8.89 (1H, s).
EXAMPLE 13: N-r3(R.S)-5-Cvclohexvl-2.3-dihvdro-1-meths1-2-oxo- 1H-1,4-benzodiazepin-3-yl]N'-3-(N-morpholinomethyl)phenyl]urea Step 1: 3(R,S)-[(Benzyloxycarbonyl)amino]-5-cyclohexyl1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one A solution of 3(R,S)-[(benzyloxycarbonyl)amino]-5-cyclohexyl- 1,3-dihydro-2H-1,4-benzodiazepin-2-one (1.1g) in dimethylformamide (13ml), under an atmosphere of nitrogen, was treated with sodium hydride (117mg of a 55-60% dispersion in mineral oil) in one portion, at -100C. After 30 min at -100C, iodomethane (174p1) was added in one portion and the solution allowed to reach 0 C over 1h. The solvent was then removed in vacuo and the crude residue partitioned between water (lOOml) and dichloromethane (100ml). The organic phase was separated and the aqueous phase extracted with dichloromethane (2 x 100ml).
The combined organic layers were washed with brine, dried (MgSO4) and evaporated. The residue was chromatographed on silica, using 1:1 petrol:ethyl acetate as the eluant, to afford the title compound (0.75g) as a white solid. Mp 205-2070C. 1H NMR (360MHz, CDCl3) # 1.03-2.04 (10H, m), 2.76 (1H, m), 3.36 (3H, s), 5.10 (3H, m), 6.52 (1H, d, J=8Hz), 7.25-7.55 (9H, m).
Step 2: 3(R,S)-Amino-5-cyclohexyl]-1,3-dihydro-1-methyl 2H-1 .4-benzodiazepin-2-one A mixture of 3(R,S)-[(benzyloxycarbonyl)amino]-5-cyclohexyl- 1,3-dihydro-1-methyl-2H-1,4-benzodiazepin-2-one (3.Og) and hydrobromic acid (45% in acetic acid, 6.2ml) was stirred for 1h at room temperature under an atmosphere nitrogen The mixture was then diluted with cold anhydrous diethyl ether (40ml) and it was stirred at 0 C for 45 min. - The white precipitate was collectedby filtration, washed with cold diethyl ether (4 x 30ml) and then dissolved in a mixture of water (30ml) and aqueous sodium hydroxide (2M, 15ml). The basic aqueous phase was extracted with ethyl acetate (3 x 70ml) and the combined organic layers were washed with brine (30ml), dried (Na2SO4) and concentrated. The residue was chromatographed on silica gel using 94:6, dichloromethane:methanol as the eluant, to afford the title compound (1.6g). Mp 133-136 C.
1H NMR (360MHz, CDCl3) # 1.02-1.40 (4H, m), 1.47-1.56 (1H, m), 1.61-1.74 (3H, m), 1.84-1.91 (1H, m), 1.96-2.06 (1H, m), 2.17 (2H, brs), 2.70-2.80 (1H, m), 3.39 (3H, s), 4.29 (1H, s), 7.20-7.27 (2H, m), 7.44-7.54 (2H, m).
Step 3: N-r3(R.S)-5-Cvclohexvl-2-3-dihvdro-1-methvl-2-oxo- 1H-1.4-benzodiazepin-3-yl]N'-[3-(N-morpholinomethyl)phenyl]urea 1-Amino-3-(N-morpholinomethyl)benzene (54mg) in anhydrous tetrahydrofuran (8ml) was cooled to OOC whereupon triphosgene (27mg) was added. The mixture was stirred at OOC for 2 min, then triethylamine (114p1) was added portionwise until pH8. The mixture was then stirred for a further 5 min, allowed to warm to 15 C, and then re-cooled to 0 C. Then a solution of 3(R,S)-amino-5-cyclohexyl-1,3-dihydro-1-methyl-2H1,4-benzodiazepin-2-one (59mg) in anhydrous tetrahydrofuran (8ml) was added dropwise over 5 min.The mixture was stirred at 0 C for 5 min, allowed to warm to room temperature and then stirred for a further 30 min. The undissolved material was removed by filtration. The solvent was evaporated in vacuo and the residue partitioned between ethyl acetate (30ml) and water (20ml). The organic phase was separated, dried (Na2SO4) and evaporated. The crude solid was recrystallised from ethyl acetate to afford the title compound as a colourless solid (50mg).
H NMR (250MHz, D6-DMSO)# 0.80-1.70-(8H, m), 1.72-1.84 (1H, m), 1.85-2.00 (1H, m), 2.24-2.39 (4H, m), 2.84-3.01 (1H, m), 3.32 (3H, s), 3.36 (2H, s), 3.49-3.63 (4H, m), 5.06 (111, d, J=1OHz), 6.85 (1H, d, J=1OHz), 7.1Q-7.3Q(3H, m), 7.32-7.40 (2H, m), 7.50-7.72 (2H, m), 7.78 (1H, d, J=8Hz), 8.98 (1H, s). TLC (silica, dichloromethane:methanol 9:1) Rf = 0.65.
EXAMPLE 14: N-[3(R)-Cyclohexyl-2,3-dihydro-1-methyl-2-oxo-1H1,4-benzodiazepin-3-yl]N'-[3-(N-morpholinomethyl)phenyl]urea 1-Amino-3-(N-morpholinomethyl)benzene (270mg) in anhydrous tetrahydrofuran (40ml) was cooled to OOC whereupon triphosgene (134mg) was added. The mixture was stirred at OOC for 2 min, then triethylamine (570p1) was added portionwise until pH8. The mixture was then stirred for a further 5 min, allowed to warm to 10 C, and then re-cooled to 0 C. Then a solution of 3(R)-amino-5-cyclohexyl- 1,3-dihydro- 1-methyl-2H- 1,4-benzodiazepin-2-one (250mg) in anhydrous tetrahydrofuran (lOml) was added dropwise over 5 min. The mixture was stirred at OOC for 5 min, allowed to warm to room temperature and then stirred for a further lh.The undissolved material was removed by filtration. The solvent was evaporated in vacuo and the residue partitioned between ethyl acetate (50ml) and water (30ml). The organic phase was separated, dried (Na2SO4) and evaporated. The crude solid was chromatographed on silica gel using a gradient elution of 0e5% methanol in dichloromethane to afford the title compound (225mg).Mp 15500 (dec.). 1H NMR (250MHz, D6-DMSO) # 0.80-1.70 (8H, m), 1.72-1.84 (1H, m), 1.85-2.00 (1H, m), 2.24-2.39 (4H, m), 2.84-3.01 (1H, m), 3.32 (3H, s), 3.36 (2H, s), 3.49-3.63 (4H, m), 5.06 (1H, d, J=1OHz), 6.85 (1H, d, J=1OHz), 7.10-7.30 (3H, m), 7.32-7.40 (2H, m), 7.50-7.72 (2H, m), 7.78 (1H, d, J=8Hz), 8.98 (1H, s).
EXAMPLE 15: N-r3(R.S)-5-Cvclohexvl-2.3-dihvdro-1-methvl-2-oxo- 1H-1.4-benzodiazepin-3-yl]N'-[4-(N-morpholinomethyl)phenyl]urea Step 1: 4-(N-Morpholinomethyl)-1-nitrobenzene A solution of morpholine (2.0g) in anhydrous acetone (100ml) was treated with potassium carbonate (4.8g) and 4-nitrobenzyl bromide (5.0g). The mixture was stirred at room temperature for 16h. The solvent was evaporated and the residue partitioned between ethyl acetate (50ml) and water (50ml). The aqueous layer was washed with ethyl acetate (2 80ml). The combined organics were dried (Na2SO4), evaporated and the residue was chromatographed on silica gel with petrol:ethyl acetate (1:1) as the eluant, to afford the title compound (3.8g). Mp 82-84 C. 1H NMR (360MHz, CDCl3) 3 2.45-2.47 (4H, m), 3.51 (2H, s), 3.67-3.77 (4H, m), 7.52 (2H, d, J=8.7Hz), 8.19 (2H, d, J=8.7Hz).
Step 2: 1-Amino-4-(N-morpholinomethyl)benzene 4-(N-Morpholinomethyl)- 1-nitrobenzene (2.00g) in ethanol (150ml) was treated with 10% palladium on carbon (200mg, 10% (w/w)). The mixture was hydrogenated at 25psi for 4 min. The catalyst was filtered off and the solvent evaporated. The residue was chromatographed on silica gel, with ethyl acetate as eluant, to afford the title compound (800mg). Mp 106-108 C. 1H NMR (250MHz, CDCl3) # 2.42-2.66 (4H, m), 3.52 (2H, s), 3.78 (2H, brs), 3.80-3.91 (4H, m), 6.72-6.84 (2H, d, J=8.4Hz), 7.16-7.28 (2H, d, J=8.4Hz).
Step 3: 5-Cyclohexyl-1,3-dihydro-1-methyl-3(R,S)-[(4nitrophenyloxycarbonyl)amino]-2H-1,4-benzodiazepin-2-one A solution of 3(R,S)-amino-5-cyclohexyl-1,3-dihydro-1-methyl- 2H-1,4-benzodiazepin-2-one (1.Og) in anhydrous tetrahydrofuran (20ml) under an atmosphere of nitrogen at OOC was treated with triethylamine (0.51ml), followed by a solution of 4-nitrophenyl chloroformate (0.75g) in anhydrous tefrahydrofuran (i;0m') dropwise. After stirring at ambient temperature for 20min, the solid which precipitated from the mixture was filtered and the filtrate was evaporated in vacuo. The residue was triturated with diethyl ether to give the title compound (1.2g, 75%) as a colourless solid.
Mp 165-168 C. 1H NMR (360MHz, CDCl3) 3 1.05 (1H, m), 1.18-1.42 (3H, m), 1.55 (1H, m), 1.65 (3H, m), 1.87 (1H, m), 2.05 (1H, m), 2.80 (1H, m), 3.43 (3H, s), 5.18 (1H, d, J=8.3Hz), 6.90 (1H, d, J=8.2Hz), 7.30 (4H, m), 7.57 (2H, m), 8.23 (2H, d, J=7.1Hz).
Step 4: N-r3(R.S)-6-Cvclohexvl-2.3-dihvdro-1-methvl-2-oXo- 1H-1,4-benzodiazepin-3-yl]N'-[4-(N-morpholinomethyl)phenyl]urea A solution of 5-cyclohexyl- 1,3-dihydro- l-methyl-3(R,S)-[(4- nitrophenyloxycarbonyl)amino]-2H- 1 ,4-benzodiazepin-2-one (200mg) in anhydrous dimethylformamide (8ml), under an atmosphere of nitrogen, was treated with triethylamine (63p1). This mixture was stirred at room temperature for 5 min whereupon a solution of 1-amino-4-(N- morpholinomethyl)benzene (88mg) in anhydrous dimethylformamide (4ml) was added dropwise over 5 min. The mixture was heated to 50 C and stirred for 5h. The solvent was evaporated and the residue partitioned between ethyl acetate (50ml) and water (50ml). The organic phase was separated, dried (Na2S04) and evaporated in vacuo.The crude solid was recrystallised from ethyl acetate to afford the title compound (75mg) as a colourless solid. Mp 170-172 C. 1H NMR (360MHz, CDCl3) 6 0.96-1.75 (8H, m), 1.77-1.90 (1H, m), 1.96-2.07 (1H, m), 2.37-2.48 (4H, m), 2.72-2.83 (1H, m), 3.40 (3H, s), 3.43 (2H, s), 3.65-3.76 (4H, m), 5.38 (1H, d, J=8.0Hz), 6.63 (1H, d, J=8.1Hz), 6.65 (1H, s), 7.20-7.34 (6H, m), 7.46-7.60 (2H, m).
EXAMPLE 16: N-r3(R.5)-5-Cvelohexvl-2.3-dihvdro-1-methvl-2-ogo- 1H-1.4-benzodiazepin-3-yl]N'-[2-(N-morpholinomethyl)phenyl]urea Step 1: 1-Amino-2-(N-morpholinomethyl)benzene A solution of morpholine (2.0g) in anhydrous acetone (100ml) was treated with potassium carbonate (4.8g) and 2-nitrobenzyl bromide (5.0g). The mixture was stirred at room temperature for 16h. The solvent was evaporated and the residue partitioned between ethyl acetate (50ml) and water (50ml). The aqueous layer was washed with ethyl acetate (2 x 50ml). The combined organics were dried (Na2SO4) and evaporated to afford crude 2-(N morpholinomethyl)- 1-nitrobenzene (3.6g).
2-(N-Morpholinomethyl)- 1-nitrobenzene (2.0g) in ethanol (150ml) was treated with 10% palladium on carbon (200mg, 10% (w/w)). The mixture was hydrogenated at 25psi for 10 min. The catalyst was filtered off and the solvent evaporated. The residue was chromatographed on silica gel with petrol (60/80):ethyl acetate (2:1) as the eluant to afford the title compound (1.29g) as colourless needles. Mp 72-73 C. 1H NMR (250MHz, CDCl3) 8 2.36-2.50 (4H, m), 3.53 (2H, s), 3.65-3.77 (4H, m), 6.62-6.73 (2H, m), 6.96-7.14 (2H, m).
Step 2: N-r3(R.S)-5-Cvclohexvl-2*3-dihvdro-1-methvl-2-oxo- 1H-1.4-benzodiazepin-3-yl[N'-[2-(N-morpholinomethyl)phenyl]urea A solution of 5-cyclohexyl-1,3-dihydro-1-methyl-3(R,S)-[(4- nitrophenyloxycarbonyl)amino]-2H-1,4-benzodiazepin-2-one (200mg) in anhydrous dimethylformamide (8ml), under an atmosphere of nitrogen, was treated with triethylamine (63p1). This mixture was stirred at room temperature for 5 min whereupon a solution of 1-amino-2-(N-morpholinomethyl)benzene (88mg) in anhydrous dimethylformamide (4ml) was added dropwise over 5 min. The mixture was heated to 500C and stirred for 3h. The solvent was evaporated and the residue was partitioned between ethyl acetate (50ml) and water (50ml).The organic phase was separated, dried (Na2SO4) and evaporated in vacuo. The crude solid was recrystallised from ethyl acetate to afford the title compound (llOmg). Mp 1680C. 1H NMR (360MHz, CDCl3) 6 1.00;1.10 (1H, m), 1.11-1.44 (3H, m), 1.50-1.75 (4H, m), 1.80-1.92 (1H, m), 2.00-2.10 (1H, m), 2.42-2.60 (4H, m), 2.73-2.86 (1H,m), 3.41 (3H, s), 3.52-3.68 (2H, m), 3.70-3.90 (4H, m), 5.39 (1H, d, J=8.1Hz), 6.33 (1H, d, J=7.9Hz), 6.91 (1H, t, J=7.2Hz), 7.06 (1H, d, J=6.8Hz), 7.20-7.34 (3H, m), 7.46-7.60 (2H, m), 7.97 (1H, d, J=8.2Hz), 9.69 (1H, s).
EXAMPLE 17: N-[3(R,S)-5-Cyclohexyl-2,3-dihydro-2-oxo-1-propyl1H-1,4-benzodiazepin-3-yl]N'-[3-(N-morpholinomethyl)phenyl]urea 1-Amino-3-(N-morpholinomethyl)benzene (236mg) in anhydrous tetrahydrofuran (40ml) was cooled to 0 C whereupon triphosgene (120mg) was added. The mixture was stirred at 0 C for 2 min, then triethylamine (5131l1) was added portionwise until pH8. The mixture was then stirred for a further 5 min, allowed to warm to 100C, and then recooled to OOC. Then a solution of 3(R,S)-amino-5-cyclohexyl-1,3-dihydro-1-propyl-2H-1,4benzodiazepin-2-one (250mg) in anhydrous tetrahydrofuran (lOml) was added dropwise over 5 min. The mixture was stirred at 0 C for 5 min, allowed to warm to room temperature and then stirred for a further 2h. The solvent was evaporated in vacuo and the residue partitioned between ethyl acetate (50ml) and water (20ml). The organic phase was separated, dried (Na2SO4) and evaporated. The crude solid was chromatographed on silica gel with dichloromethane:methanol (99:1) as the eluant to afford the title compound (216mg). Mp 230 C. 1H NMR (360MHz, CDCl3) 6 0.82 (3H, t, J=7.4Hz), 1.00-1.90 (11H, m), 1.99-2.10 (1H, m), 2.40-2.52 (4H, m), 2.74-2.86 (1H, m), 3.46 (2H, s), 3.50-3.62 (1H, m), 3.66-3.76 (4H, m), 4.26-4.38 (1H, m), 5.33 (1H, d, J=7.9Hz), 6.66 (1H, d, J=8.0Hz), 6.83 (1H, s), 7.00 (1H, d, J=6.5Hz), 7.16-7.42 (5H, m), 7.45-7.60 (2H, m).
EXAMPLE 18: N-[3(R,S)-5-Cycloheptyl-2,3-dihydro-1-methyl-2 oxo- lH- .4-benzodiazepin-3-vllN'-r3-(N-methvl-N'-piperazinvl) phenyl]urea Step 1: 5-Cycloheptyl-1,3-dihydro-1-methyl-3(R,S)-[(4 nitrophenvloxvcarbonvlEaminol-2H-1.4-benzodiazepin-2-one 3(R,S)-[Benzyloxycarbonyl)amino]-5-cycloheptyl-1,3dihydro-1-methyl-2H-1,4-benzodiazepin-2-one (0.67g) was dissolved in hydrobromic acid (4ml of a 30% solution in glacial acetic acid) and stirred at room temperature for 20min. The yellow solution was then added dropwise to anhydrous diethyl ether (20ml) at OOC and the resultant cream solid filtered off and washed with ether.
The solid was partitioned between dichloromethane (some) and 10% sodium hydroxide solution (501). The organic layer was separated, dried (Na2SO4) and evaporated in vacuo to afford crude 3(R, S)-amino-5-cycloheptyl- 1,3-dihydro- l-methyl-2H-1,4- benzodiazepin-2-one as a colourless viscous oil.
The amine was dissolved in anhydrous tetrahydrofuran (9ml) under an atmosphere of nitrogen, at room temperature, and triethylamine (221p1) was added dropwise. A solution of 4-nitrophenyl chloroformate (322mg) in anhydrous tetrahydrofuran (9ml) was then added, and the reaction mixture stirred for a further 3h. After this time the undissolved solid was filtered off, washed with tetrahydrofuran, and the filtrate evaporated in vacuo. The residue was azeotroped with toluene (2 x 50ml) and the residue triturated with anhydrous ether to afford the title compound (425mg) as a white solid. TLC (silica, petrol (60/80):ethyl acetate 2:1).Rf= 0.3. 1H NMR (360MHz, CDCl3) 8 1.22-2.10 (12H, m), 2.98 (1H, m), 3.45 (3H, s), 5.15 (1H, d, J=8.2Hz), 6.89 (1H, d, J=9.2Hz), 7.26-7.35 (4H, m), 7.49-7.60 (2H, m), 8.22 (2H, d, J=7.1Hz).
Step 2: N-[3(R,S)-5-Cycloheptyl-2,3-dihydro-1-methyl-2-oxo-1H1,4-benzodiazepin-3-yl]N'-[3-(N-methyl-N'-piperazinyl)phenyl]urea To a stirred solution of 5-cycloheptyl-1,3-dihydro-1-methyl3(R,S)-[(4-nitrophenyloxycarbonyl)amino]-2H-1,4-benzodiazepin-2one (200mg) in anhydrous dimethylformafriide (5ml), under an atmosphere of nitrogen, was added triethylamine (61 l). After 2 min a solution of 1-amino-3-(N-methyl-N'-piperazinyl)benzene (85mg) in anhydrous dimethylformamide (5ml) was added and the solution heated at 50 C for 1.5h. After this time the solvent was evaporated in vacuo and the residue partitioned between ethyl acetate (20ml) and water (20ml). An undissolved solid was filtered off and triturated with anhydrous diethyl ether to give the desired urea (128mg) as a white solid. 1H NMR (360MHz, D6-DMSO) 61.13-1.94 (12H, m), 2.19 (3H, s), 2.40-2.43 (4H, m), 3.03-3.12 (5H, m), 3.30 (3H, s), 5.04 (1H, d, J=8.4Hz), 6.49 (1H, dd, 8.1 and 1Hz), 6.62 (1H, d, J=9.0Hz), 7.03 (1H, t, J=8.1Hz), 7.12 (1H, m), 7.22 (1H, d, J=8.4Hz), 7.38 (1H, dd, J=7.6 and 7.6Hz), 7.54 (1H, d, J=7.7Hz), 7.63 (1H, dd, J=8.3 and 8.3Hz), 7.76 (1H, d, J=7.6Hz), 8.86 (1H, s).
MS (CI, NH3) 502 (M+).

Claims (9)

CLAIMS:
1. A compound of the formula (I):
wherein: R represents H, C1-6alkyl, C3-7-cycloalkyl, cyclopropylmethyl, CH2Co2R5 (where R5 is C14alky1), CH2CONR6R7 (where R6 and R7 each independently represents H or C1-4alkyl, or R6 and R7 together form a chain (CH2)p where p is 4 or 5), C1-6alkylNR8R9 or C1-6alkylCONR8R9 where R8 and R9 together with the nitrogen atom to which they are attached form a 5 to 8 membered non-aromatic ring system containing a second heteroatom selected from O, S or NR10, where R12 is H or C1-4alkyl;; R2 represents a phenyl or pyridyl group optionally substituted by one or more substituents salecdted from C1-6alkyl, halc, hydroxy, C1-4alkoxy, (CH2)q-tetrazolyl optionally substituted in the tetrazole ring by C1-4alkyl, (CH2)q-imidazolyl, (CH2)q-triazolyl (where q is 0, 1, 2 or 3), 5-hydroxy-4-pyrone, NR6R7, NR10COR5, NR10CONR10,R5 (where R10 and R10' are each independently H or C1-4alkyl and R5 is as previously defined), CONR6R7 (where R6 and R7 are as previously defined), SO(Cl~6alkyl), SO2(C1-6alkyl), trifluoromethyl, CONHSO2R11, SO2NHCOR11 (where R11 is C1-6alkyl, optionally substituted aryl, 2,2-difluorocyclopropane or trifluoromethyl), SO2NHR12 (where R12 is a nitrogen containing heterocycle), B(OH)2 or (CH2)qCO2H, where q is as previously defined; or R2 represents a group
where X1 represents CH or N; W represents CH2 or NR10, where R10 is as previously defined, and W1 represents CH2, or W and W1 each represent 0; or R2 represents phenyl substituted by a group
wherein x2 is 0, S or NR10, where R10 is as previously defined;Z is a bond, 0 or S; m is 1, 2 or 3; n is 1, 2 or 3; and y is 0, 1, 2 or 3; Each R3 represents C1-6alkyl, halo or NR6R7, where R6 and R7 are as previously defined; R4 represents bridged C6-10bicycloalkyl or C37cycloalkyl optionally substituted by one or more C1-4alkyl groups; x is 0, 1, 2 or 3; with the proviso that when R1 is H, C1-6alkyl, C3-7cycloalkyl, cyclopropylmethyl, CH2CO2R5 or CH2CONR6R7, R2 represents phenyl substituted by
and salts and prodrugs thereof.
2. A compound as claimed in claim 1 wherein R1 is C1-6alkyl, C3-7 cycloalkyl, cyclopropylmethyl, CH2CO2R5 or CH2CONR6R7; R is phenyl substituted by a group:
wherein X2, Z, m, n and y are as previously defined; and R4 represents C3-7cycloalkyl.
3. A compound as claimed in claim 1 wherein R1 is C16 alkyl NR8R9 or C16 alkyl CONR8R9; R2 is a phenyl group optionally substituted by one or more of C1-6alkyl, halo, hydroxy, C1-4alkoxy, (CH2)q-tetrazolyl optionally substituted in the tetrazole ring by C1-4alkyl, (CH2)q imidazolyl, (CH2)q-triazolyl, 5-hydroxy-4-pyrone, NR6R7, CONR6R7, NR10COR, NR10CONR10'R5, SO(C1-6alkyl), SO2(C1 6alkyl), trifluoromethyl, CONHSO2R, SO2NHCOR, SO2NHCOR1l, SO2NHR12, B(OH)2, (CH2)qCO2H; or R2 represents a group
where W and W' are as previously defined.
4. A compound of the formula (Ia):
wherein R20 is C4-7cycloalkyl; R21 is C1-4alkyl and R22 is H, or R21 and R22 together form a chain (CH2)3; n is 2 or 3; and salts and prodrugs thereof.
5. A compound of the formula (Ib):
wherein X is as defined for formula (I); R is C1-6alkyl; R24 is cyclopentyl, cyclohexyl or cycloheptyl; z is 0 or 1; and salts and prodrugs thereof.
6. A compound selected from N-[3(R,S)-5-Cycloheptyl-2,3-dihydro-1-(2-(N- morpholino) ethyl)-2-oxo-1H-1,4-benzodiazepin-3-yl] N-|-(3methylphenyl] urea N-[3(R,S)-5-Cyclohexyl-2,3-dihydro-1-(2-(N morpholino)ethyl)-2-oxo-1H-1,4-benzodiazepin-3#yl]N-|-[3- methylphenyl] urea N-[3-(R,S)-5-Cyclohexyl-2,3-dihydro-1-(2-(Nmorpholino)ethyl)-2-oxo-1H-1,4-benzodiazepin-3-yl] N-|-[5indanyl] urea N-[3(R,S)-5-cyclohexyl-2,3-dihydro-2-oxo-1 propyl-1H-1,4-benzodiazepin-3#yl] N-|-[4-(Npiperazinyl)phenyl]urea N-[3(R,S)-5-Cyclohexyl-2,3-dihydro-2-oxo-1- propyl-1H-1,4-benzodiazepin-3-yl] N-|-[4-(N-methyl-N-|piperazinyl)phenyl]urea N-[3(R,S)-5-Cyclohexyl-2,3-dihydro-2-oxo-1propyl-1H-1,4-benzodiazepin-3-yl]N-|-[3-(N-methyl-N-|piperazinyl)phenyl]urea N-[3(R,S)-2,3-Dihydro-1-methyl-5-(4 methylpiperidin-l-yl)-2-oxo-lH-1,4-benzodiazepin-3-yl]Nd- [3-(N-methyl-N-|-piperazinyl)phenyl]urea N-[3(R,S)-5-Cycloheptyl-2,3-dihydro-1-methyl-2oxo-1H-1,4-benzodiazepin-3-yl]N-|-[3-(Nmorpholinomethyl)phenyl] urea N-(R,S)-5-Cycloheptyl-2,3-dihydro-1-methyl-2oxo-1H-1,4-benzodiazepin-3-yl]N-|-[3-(2-(Nmorpholino)ethoxy)phenyl]urea N-[3(R)-5-Cyclohexyl-2,3-dihydro-1-methyl-2oxo-1H-1,4-benzodiazepin-3-yl]N-|-[3-(2-(Nmorpholino)ethoxy)phenyl]urea N-[3(R,S)-5-Cycloheptyl-2,3-dihydro-1 (2#methylpropyl)-2-oxo-1H,1,4-benzodiazepin-3-yl]N-|-[3 (N-methyl-N-piperazinyl) phenyl)urea (+)-N-[5-Cycloheptyl-2,3-dihydro-1-methyl-2oxo-lH-1,4- benzodiazepin-3-yl] N-[3-(N-methyl-N- piperazinyl)phenyl]urea N-[3(R,S)-5-Cyclohexyl-2,3-dihydro-1-methyl-2- oxo-1H-1,4-benzodiazepin-3-yl]N-|-3-(Nmorpholinomethyl)phenyl]urea N-[3(R)-Cyclohexyl-2,3-dihydro-1-methyl-2-oxo1H-1,4-benzodiazepin-3-yl]N-|-[3-(Nmorpholinomethyl)phenyl]urea N-[3(R,S)-5-Ctyclohexyl-2,3-dihydro-1-methyl-2oxo-1H-1,4-benzodiazepin-3-yl]N-|-[4-(Nmorpholinomethyl)phenyl]urea N-[3(R,S)-5-Cyclohexyl-2,3-dihydro-1-methyl-2 oxo-1H-1,4-Senzodiqzepin-37yl]N-1-C2-(N- morpholinomethyl)phenyl]urea N-[3(R,S)-5-Cyclohexyl-2,3-dihydro-2-oxo-1propyl-1H-1,4-benzodiazepin-3-yl]N-|-[3-(Nmorpholinomethyl)phenyl]urea N-t3(R,S)-5-Cycloheptyl-2,3-dihydro-1-methyl-2- oxo-1H-1,4-benzodiazepin-3-yl]N-|-[3-(N-methyl-N-|piperazinyl) phenyl]urea or pharmaceutically acceptable salts thereof.
7. The use of a compound as claimed in any of claims 1 to 6 in therapy.
8. A compound as claimed in any of claims 1 to 6 for use in the preparation of a medicament.
9. A pharmaceutical composition which comprises a compound of the formula (I) as defined in claim 1 or a pharmaceutically acceptable salt thereof in association with a carrier therefor.
GB9322017A 1992-11-02 1993-10-26 N-(2-oxo-1H-1,4-benzodiazepin-3-yl)-ureas Withdrawn GB2271991A (en)

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GB929222934A GB9222934D0 (en) 1992-11-02 1992-11-02 Therapeutic compounds
GB929223584A GB9223584D0 (en) 1992-11-11 1992-11-11 Therapeutic agents
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8889730B2 (en) 2012-04-10 2014-11-18 Pfizer Inc. Indole and indazole compounds that activate AMPK
US9394285B2 (en) 2013-03-15 2016-07-19 Pfizer Inc. Indole and indazole compounds that activate AMPK

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0514133A1 (en) * 1991-05-14 1992-11-19 MERCK SHARP &amp; DOHME LTD. Benzodiazepine derivatives, compositions containing them and their use in therapy

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0514133A1 (en) * 1991-05-14 1992-11-19 MERCK SHARP &amp; DOHME LTD. Benzodiazepine derivatives, compositions containing them and their use in therapy

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8889730B2 (en) 2012-04-10 2014-11-18 Pfizer Inc. Indole and indazole compounds that activate AMPK
US9394285B2 (en) 2013-03-15 2016-07-19 Pfizer Inc. Indole and indazole compounds that activate AMPK

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