WO2006070292A2 - Composes de pyrimidine en tant qu'agents anti-ictogeniques et/ou agents anti-epileptogeniques - Google Patents

Composes de pyrimidine en tant qu'agents anti-ictogeniques et/ou agents anti-epileptogeniques Download PDF

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WO2006070292A2
WO2006070292A2 PCT/IB2005/004156 IB2005004156W WO2006070292A2 WO 2006070292 A2 WO2006070292 A2 WO 2006070292A2 IB 2005004156 W IB2005004156 W IB 2005004156W WO 2006070292 A2 WO2006070292 A2 WO 2006070292A2
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benzyl
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phenyl
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Donald F. Weaver
Buhendwa Musole Guillain
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Queen's University
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
    • C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
    • C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
    • C07D239/46—Two or more oxygen, sulphur or nitrogen atoms
    • C07D239/52—Two oxygen atoms
    • C07D239/54—Two oxygen atoms as doubly bound oxygen atoms or as unsubstituted hydroxy radicals
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
    • C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
    • C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
    • C07D239/46—Two or more oxygen, sulphur or nitrogen atoms
    • C07D239/52—Two oxygen atoms
    • C07D239/54—Two oxygen atoms as doubly bound oxygen atoms or as unsubstituted hydroxy radicals
    • C07D239/545—Two oxygen atoms as doubly bound oxygen atoms or as unsubstituted hydroxy radicals with other hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
    • C07D239/553—Two oxygen atoms as doubly bound oxygen atoms or as unsubstituted hydroxy radicals with other hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms with halogen atoms or nitro radicals directly attached to ring carbon atoms, e.g. fluorouracil
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
    • C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
    • C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
    • C07D239/46—Two or more oxygen, sulphur or nitrogen atoms
    • C07D239/56—One oxygen atom and one sulfur atom
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/04—Ortho-condensed systems

Definitions

  • Epilepsy is a serious neurological condition, associated with seizures, that affects hundreds of thousands of people worldwide.
  • a seizure results from a sudden electrical discharge from a collection of neurons in the brain.
  • the resulting nerve cell activity is manifested by symptoms such as uncontrollable movements.
  • a seizure is a single discrete clinical event caused by an excessive electrical discharge from a collection of neurons through a process termed "ictogenesis.”
  • ictogenesis a seizure is merely the symptom of epilepsy.
  • epilepsy is a dynamic and often progressive process, characterized by an underlying sequence of pathological transformations whereby normal brain is altered, becoming susceptible to recurrent seizures through a process termed "epileptogenesis.” While it is believed that ictogenesis and epileptogenesis have certain biochemical pathways in common, the two processes are not identical.
  • Ictogenesis (which may be described as the initiation and propagation of a seizure in time and space) is a rapid and definitive electrical/chemical event occurring over seconds or minutes, hi contrast, epileptogenesis (which may be explained as the gradual process whereby normal brain is transformed into a state susceptible to spontaneous, episodic, time-limited, recurrent seizures, through the initiation and maturation of an "epileptogenic focus") is a slow biochemical and/or histological process which generally occurs over months to years. Epileptogenesis is a two phase process.
  • Phase One epileptogenesis also known as primary epileptogenesis, characterizes the initiation of the epileptogenic process prior to the first seizure, and is often the result of stroke, disease (e.g., meningitis), or trauma, such as an accidental blow to the head or a surgical procedure performed on the brain.
  • Phase Two epileptogenesis also known as secondary epileptogenesis, refers to the process during which an individual already susceptible to seizures, becomes still more susceptible to seizures of increasing frequency and/or severity.
  • NMDA N-methyl-D-aspartate
  • Other researchers implicate downregulation of inhibitory coupling between neurons, mediated by gamma- amino-butyric acid (GABA) receptors.
  • GABA gamma- amino-butyric acid
  • epileptic seizures are rarely fatal, large numbers of patients require medication to avoid the disruptive, and potentially dangerous, consequences of seizures.
  • medication is required for extended periods of time, and in some cases, a patient must continue to take prescription drugs for life.
  • drugs used for the management of epilepsy have side effects associated with prolonged usage, and the cost of the drugs can be considerable.
  • anticonvulsant agents such as phenytoin, valproate and carbamazepine (ion channel blockers), as well as newer agents like felbamate, gabapentin, and tiagabine.
  • anticonvulsant agents such as phenytoin, valproate and carbamazepine (ion channel blockers)
  • newer agents like felbamate, gabapentin, and tiagabine.
  • anticonvulsant is synonymous with “antiseizure” or “anti-ictogenic.” While these drugs can suppress seizures by blocking ictogenesis, it is believed that they do not influence the development of epilepsy because they do not block epileptogenesis.
  • VNS vagus nerve stimulation
  • the present invention is directed to the development of uracil-based compounds which may be used to treat, prevent, or inhibit seizure-associated states, e.g., ranging from discrete seizure episodes to dynamic pathological transformations leading to susceptibility to recurrent seizures to epilepsy.
  • seizure-associated states e.g., ranging from discrete seizure episodes to dynamic pathological transformations leading to susceptibility to recurrent seizures to epilepsy.
  • Such compounds are capable of treating, preventing, or inhibiting the seizure-associated state directly ⁇ e.g., as an anti- ictogenic agent or an anti-epileptogenic agent), or converting in vivo to an active agent, or acting through a combination thereof.
  • the present invention is also directed to new synthetic methods, for example, methods allowing production of l-alkyl-5-bromouracil and thiouracil analogs, as well as 5,5'-biuracil analogs.
  • the invention is directed to a compound, e.g., an anti-ictogenic or anti-epileptogenic compound, of the formula
  • X is S or O
  • R 1 is selected from the group consisting of H, benzyl, isopropyl, ⁇ -NO 2 -benzyl, benzoyl, and -CH 2 SCH 3 ;
  • R 2 is selected from the group consisting of H, CH 3 ,
  • R 3 is selected from the group consisting of H, Br, I, CH 3 , -C ⁇ CC 4 H 9 , -CH 2 N(H)-phenyl, -CH 2 N(H)CH 2 -phenyl, -CH 2 -O-benzyl, an alkylsulfide, and an alkyluracil; and
  • R 4 is selected from the group consisting of H, benzyl, isopropyl, p-NO 2 -benzyl, and benzoyl; or pharmaceutically acceptable salts or esters thereof.
  • the invention pertains to a pharmaceutical composition
  • a pharmaceutical composition comprising a sufficient amount of a compound of the present invention to treat, prevent, or inhibit a seizure-associated state , e.g., epilepsy, in a subject in need thereof, and a pharmaceutically acceptable carrier.
  • the pharmaceutical composition comprises a compound that is anti-ictogenic and/or anti-epileptogenic.
  • the invention in another aspect, relates to a method of treating, preventing, or inhibiting a seizure-associated state in a subject comprising administering to a subject in need thereof a sufficient amount of a compound of the present invention, such that the seizure-associated state is treated, prevented, or inhibited.
  • An additional aspect of the invention is a method of treating, preventing, or inhibiting ictogenesis in a subject comprising administering to a subject in need thereof a sufficient amount of a compound of the present invention, such that ictogenesis is treated, prevented, or inhibited.
  • Yet another aspect of the invention relates to a method of treating, preventing, or inhibiting epileptogenesis in a subject comprising administering to a subject a sufficient amount of a compound of the present invention, such that epileptogenesis is treated or prevented.
  • the invention is directed to a kit for treating, preventing, or inhibiting a seizure-associated state in a subject comprising a compound of the present invention, and instructions for administering a sufficient amount of the compound to a subject, such that the seizure-associated state is treated, prevented, or inhibited.
  • the invention is directed to a packaged pharmaceutical composition for treating, preventing, or inhibiting a seizure-associated state in a subject comprising a container holding a compound of the present invention; and instructions for using the compound for treating, preventing, or inhibiting the seizure-associated state in a subject.
  • the invention is directed to a kit for treating, preventing, or inhibiting ictogenesis in a subject comprising a compound of the present invention, and instructions for administering a sufficient amount of the compound to a subject, such that ictogenesis is treated, prevented, or inhibited.
  • Another aspect of the invention is a packaged pharmaceutical composition for treating, preventing, or inhibiting ictogenesis in a subject comprising a container holding a compound of the present invention; and instructions for using the compound for treating, preventing, or inhibiting ictogenesis in a subject.
  • the invention pertains to a kit for treating, preventing, or inhibiting epileptogenesis in a subject comprising a compound of the present invention, and instructions for administering a sufficient amount of the compound to a subject, such that epileptogenesis is treated, prevented, or inhibited.
  • the invention is directed to a packaged pharmaceutical composition for treating, preventing, or inhibiting epileptogenesis in a subject comprising a container holding a compound of the present invention; and instructions for using the compound for treating, preventing, or inhibiting epileptogenesis in a subject.
  • the invention is a method of treating, preventing, or inhibiting seizures, e.g., seizures caused by epilepsy, in a subject suffering from head trauma comprising administering to a subject a sufficient amount of a compound of the present invention, such that seizures are treated, prevented, or inhibited.
  • seizures e.g., seizures caused by epilepsy
  • the invention is a method of treating, preventing, or inhibiting epileptogenesis in a subject suffering from head trauma comprising administering to a subject a sufficient amount of a compound of the present invention, such that epileptogenesis is treated, prevented, or inhibited.
  • Another aspect of the invention pertains to a method of treating, preventing, or inhibiting seizures in a subject suffering from traumatic brain injury (TBI) comprising administering to a subject a sufficient amount of a compound of the present invention, such that seizures are treated, prevented, or inhibited.
  • TBI traumatic brain injury
  • Another aspect of the invention pertains to a method of treating, preventing, or inhibiting epileptogenesis in a subject suffering from traumatic brain injury (TBI) comprising administering to a subject a sufficient amount of a compound of the present invention, such that epileptogenesis is treated, prevented, or inhibited.
  • TBI traumatic brain injury
  • Another aspect of the invention pertains to a compound of the formula
  • X is S or O;
  • R 1 is selected from the group consisting of H, benzyl, isopropyl, p-NO 2 -benzyl, benzoyl, and -CH 2 SCH 3 ;
  • R 2 is selected from the group consisting of H, CH 3 , -CH 2 CH 2 -phenyl, n-butyl, ethyl, n-propyl, phenyl, benzyl, and -CH 2 CH(OH)-phenyl;
  • R 3 is selected from the group consisting of H, Br, I, CH 3 , -C ⁇ CC 4 H 9 , -CH 2 N(H)-phenyl, -CH 2 N(H)CH 2 -phenyl, -CH 2 -O-benzyl, an alkylsulfide, and an alkyluracil; and
  • R 4 is selected from the group consisting of H, benzyl, isopropyl, p-NO 2 -benzyl, and benzoyl; or acceptable salts, e.g., pharmaceutically acceptable salts, or esters thereof.
  • the invention is directed to a method of preparing a compound of present invention, as described herein, e.g., in the Examples..
  • This invention pertains to methods and agents useful for the treatment, prevention, or inhibition of seizure-associated states; for treatment, prevention, or inhibition of epileptogenesis and/or for treatment, prevention, or inhibition of ictogenesis; and to methods for preparing compounds that are useful for the treatment, prevention, or inhibition of seizure-associated states, e.g., anti-ictogenic and/or anti- epileptogenic agents.
  • the invention further pertains to pharmaceutical compositions for treatment, prevention, or inhibition of seizure-associated states, as well as to kits including the compounds of the invention.
  • a sufficient amount of a compound is an amount of a compound effective, necessary, or sufficient to treat, prevent, or inhibit a seizure-associated state in a subject, e.g., a therapeutically effective amount of a compound.
  • the amount can vary depending on such factors as the size and weight of the subject, the type of illness, or the particular compound. For example, the choice of the compound can affect what constitutes a "sufficient amount”.
  • anionic group refers to a group that is negatively charged at physiological pH.
  • anionic groups include carboxylate, sulfate, sulfonate, sulfmate, sulfamate, tetrazolyl, phosphate, phosphonate, phosphinate, or phosphorothioate or functional equivalents thereof.
  • a particularly preferred anionic group is a carboxylate.
  • "Functional equivalents" of anionic groups include bioisosteres, e.g., bioisosteres of a carboxylate group. Bioisosteres encompass both classical bioisosteric equivalents and non-classical bioisosteric equivalents. Classical and non- classical bioisosteres are known in the art. See, e.g., Silverman, R.B. The Organic Chemistry of Drug Design and Drug Action, Academic Press, Inc.: San Diego, CA, 1992, pp. 19-23.
  • anticonvulsant agent or "anti-ictogenic agent” includes agents which are capable of reducing, preventing or stopping of ictogenesis, inhibiting or slowing down of further ictogenesis in a subject with epilepsy or a seizure-associated state, and reducing ictogenesis in a subject with ongoing epilepsy or a seizure-associated state.
  • the extent of the inhibition of ictogenesis may be a range from substantially complete elimination of ictogenesis, for example, to reduction of ictogenesis.
  • Inhibition of ictogenesis is determined relative to an untreated subject, or relative to the treated subject prior to treatment, e.g., determined by clinically measurable improvement in a biological function in a patient.
  • ictogenesis may be inhibited by the compounds of the invention as described herein.
  • anti-epileptogenic agent includes agents which are capable of inhibiting epileptogenesis when administered to a subject.
  • a process in a pathway associated with epileptogenesis includes biochemical processes or events which take place during Phase One or Phase Two of epileptogenesis and lead to epileptogenic changes in tissue, i.e., in tissues of the central nervous system (CNS), e.g., the brain. Examples of processes in pathways associated with epileptogenesis are discussed in more detail, infi-a.
  • a disorder associated with NMDA receptor activity includes disorders of a subject where abnormal ⁇ e.g., excessive) activity of NMDA receptors can be treated by antagonism of an NMDA receptor.
  • epilepsy is a disorder associated with excessive NMDA-mediated activity.
  • disorders associated with excessive NMDA-mediated activity include pain, stroke, anxiety (e.g., schizophrenia), cerebral ischemia, Huntington's chorea, Parkinson's disease, motor neuron disease (e.g, ALS), Alzheimer's disease, dementias (e.g., AIDS dementia and vascular dementia), and other disorders (in humans or animals) where excessive activity of NMDA receptors is a cause, at least in part, of the disorder.
  • ⁇ -amino anionic compound includes compounds having an amino group, such as -NR a R b , where R a and R b may each independently be hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, or aryloxycarbonyl, or R a and R b , taken together with the nitrogen atom to which they are attached, form a cyclic moiety having from 3 to 8 atoms in the ring separated from an anionic group by a two-carbon spacer unit.
  • a ⁇ -amino anionic compound can be represented by the substructural formula A-C-C-NR a R b , where A is an anionic group.
  • Preferred ⁇ -amino anionic compounds include ⁇ -amino acids and analogs thereof.
  • the ⁇ -amino anionic compound is not ⁇ - alanine or taurine.
  • the term "container” includes any receptacle for holding the therapeutic formulation.
  • the container is the packaging that contains the formulation.
  • the container is not the packaging that contains the formulation, i.e., the container is a receptacle, such as a box or vial that contains the packaged formulation or unpackaged formulation and the instructions for use of the formulation.
  • packaging techniques are well known in the art. It should be understood that the instructions for use of the therapeutic formulation may be contained on the packaging containing the therapeutic formulation, and as such the instructions form an increased functional relationship to the packaged product. However, it should be understood that the instructions can contain information pertaining to the compound's ability to perform its intended function, e.g., treat, prevent, or inhibit ictogenesis and/or epileptogenesis.
  • epilepsy describes the clinical state associated with recurrent seizures whatever the etiology or causation.
  • inhibition of epileptogenesis includes reducing, preventing or stopping of epileptogenesis, inhibiting or slowing down of further epileptogenesis in a subject with epilepsy, and reducing or reversing epileptogenesis in a subject with ongoing epilepsy.
  • the extent of the inhibition of epileptogenesis may be a range, from substantially complete elimination of epileptogenesis for example, to reduction of epileptogenesis.
  • Inhibition of epileptogenesis may be determined relative to an untreated subject, or relative to the treated subject prior to treatment, or, e.g., determined by clinically measurable improvement in a biological function in a patient or by indication that the biological function in the patient does not worsen (i.e., the patient does not develop epilepsy).
  • epileptogenesis may be inhibited by the compounds of the invention as described herein.
  • pharmacophore is known in the art, and includes molecular moieties capable of exerting a selected biochemical effect, e.g., inhibition of an enzyme, binding to a receptor, chelation of an ion, and the like.
  • a selected pharmacophore can have more than one biochemical effect, e.g. , can be an inhibitor of one enzyme and an agonist of a second enzyme.
  • a therapeutic agent can include one or more pharmacophores, which can have the same or different biochemical activities.
  • pharmacophores with similar structures and/or properties (e.g., biological effects) may be combined to predict or design an optimized or "average pharmacophore" structure. Such an average pharmacophore structure may provide a greater level of desired biological effect than the individual pharmacophores used to create the average structure.
  • seizure-associated state includes clinical states or processes which are characterized by one or more discrete seizure events or episodes caused by an excessive electrical discharge from a collection of neurons.
  • Seizure-associated states include, for example, a range from single discrete seizure episodes, e.g., occurring through ictogenesis, to dynamic pathological transformations leading to susceptibility to recurrent seizures, e.g., occurring through epileptogenesis, to epilepsy.
  • the language “seizure-associated state” is also intended to include states prior to clinical diagnosis of seizures or epilepsy, in subjects who are at risk to develop seizures or epilepsy, for example, who have suffered from head trauma or a traumatic brain injury (TBI).
  • TBI traumatic brain injury
  • the clinical state of a subject in which seizures are induced is also considered to be a seizure-associated state.
  • the seizure-associated state is ictogenesis, epileptogenesis, or epilepsy.
  • subject includes living organisms in which a seizure associated-state, e.g. , epilepsy, can occur, or which are at risk for having a seizure, e.g. , partial seizures, generalized seizures, unclassified epileptic seizures.
  • subjects include animals, such as mammals, including but not limited to cats, dogs, horses, pigs, cows, goats, sheep, rodents (e.g., mice or rats), rabbits, squirrels, bears, and primates (e.g., chimpanzees, monkeys, gorillas, apes, and humans); as well as chickens, ducks, and geese; and transgenic species thereof.
  • the subject is human or dog. In a particular embodiment, the subject is human.
  • treatment includes the application or administration of a composition of the invention to a subject, or application or administration of a composition of the invention to a cell or tissue from a subject that suffers from a seizure-associated state, or has a symptom of such a disease or condition, or is at risk of having (or susceptible to) a seizure or developing epilepsy, with the purpose of curing, healing, alleviating, relieving, altering, remedying, ameliorating, improving, or affecting the disease or condition, the symptom of the disease or condition, or the risk of (or susceptibility to) the disease or condition.
  • treating refers to any indicia of success in the treatment or amelioration of an injury, pathology or condition, including any objective or subjective parameter such as abatement; remission; diminishing of symptoms or making the injury, pathology or condition more tolerable to the subject; slowing in the rate of degeneration or decline; making the final point of degeneration less debilitating; improving a subject's physical or mental well-being; or, in some situations, preventing the onset of particular conditions.
  • the treatment or amelioration of symptoms can be based on objective or subjective parameters, including the results of a physical examination, a psychiatric evaluation, or a cognition test such as Clinical Dementia Rating (CDR) scale, Mini- mental State Examination (MMSE), Alzheimer's Disease Assessment Scale-Cognitive Subscale (ADAS-Cog), or another test known in the art.
  • CDR Clinical Dementia Rating
  • MMSE Mini- mental State Examination
  • ADAS-Cog Alzheimer's Disease Assessment Scale-Cognitive Subscale
  • the subject is in need of treatment by the methods of the invention, and is selected for treatment based on this need.
  • a subject in need of treatment is art-recognized, and includes subjects that have been identified as having a seizure-associated state, having a symptom of a seizure-associated state, or at risk of a seizure-associated state, e.g.
  • diagnosis e.g., medical diagnosis
  • treatment e.g., curing, healing, preventing, alleviating, relieving, altering, remedying, ameliorating, improving, or affecting the seizure- associated state, the symptom of the seizure-associated state, or the risk of the seizure- associated state.
  • the chemical groups of the present invention may be substituted or unsubstituted. Further, unless specifically indicated, the chemical substituents may in turn be substituted or unsubstituted. In addition, multiple substituents may be present on a chemical group or substituent.
  • substituents include alkyl, alkenyl, alkynyl, aryl, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxyl, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, formyl, trimethylsilyl, phosphate, phosphonato, phosphinato, cyano, amino (including alkyl amino, dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amido, imino, sulfhydryl, alkylthio, arylthi
  • alkyl refers to saturated aliphatic groups, including straight-chain alkyl groups, branched-chain alkyl groups, cycloalkyl, heterocyclyl, cycloalkyl (alicyclic) groups, alkyl substituted cycloalkyl groups, and cycloalkyl substituted alkyl groups.
  • a straight chain or branched chain alkyl has 30 or fewer carbon atoms in its backbone (e.g., Ci-C 3O for straight chain, C 3 -C 30 for branched chain), and more preferably has 20 or fewer carbon atoms in the backbone.
  • preferred cycloalkyls have from 4-10 carbon atoms in their ring structure, and more preferably have 5, 6 or 7 carbons in the ring structure.
  • alkyl e.g. , methyl, ethyl, propyl, butyl, pentyl, hexyl, etc.
  • alkyl include both "unsubstituted alkyl” and "substituted alkyl", the latter of which refers to alkyl moieties having substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone, which allow the molecule to perform its intended function.
  • substituents which are not intended to be limiting, include moieties selected from straight or branched alkyl (preferably C 1 -C 5 ), cycloalkyl (preferably C 3 -C 8 ), alkoxy (preferably C 1 -C 6 ), thioalkyl (preferably C 1 -C 6 ), alkenyl (preferably C 2 -C 6 ), alkynyl (preferably C 2 -C 6 ), heterocyclic, carbocyclic, aryl (e.g., phenyl), aryloxy (e.g., phenoxy), aralkyl (e.g., benzyl), aryloxyalkyl (e.g., phenyloxyalkyl), arylacetamidoyl, alkylaryl, heteroaralkyl, alkylcarbonyl and arylcarbonyl or other such acyl group, heteroarylcarbonyl, or heteroaryl group, (CR'R") 0-3 NR'R
  • substituents can include, for example, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato, cyano, amino (including alkyl amino, dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates, sulfonate, sulfamoyl, sulfonamido, nitro, trifluoromethyl
  • an "aralkyl” moiety is an alkyl substituted with an aryl (e.g., phenylmethyl (i.e., benzyl)).
  • An "alkylsulfide” moiety is alkyl substituted with a substituted sulfur group, e.g., substituted with a methyl or benzyl, i.e., forming a sulfide or thioether.
  • alkylsulfide also includes alkylsulfides in different oxidation states, e.g., which may further be substituted with one or more oxygens attached by single or double bonds, e.g., sulfoxide or sulfone.
  • An "alkyluracil” moiety is an alkyl substituted with a uracil group. It will be understood that “substitution” or “substituted with” includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc.
  • substituted includes all permissible substituents of organic compounds.
  • permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds.
  • aryl includes 5- and 6-membered single-ring aromatic groups, that may include from zero to four heteroatoms, for example, benzene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine, and the like.
  • Aryl groups also include polycyclic fused aromatic groups such as naphthyl, quinolyl, indolyl, and the like.
  • aryl groups having heteroatoms in the ring structure may also be referred to as "aryl heterocycles," “heteroaryls” or “heteroaromatics.”
  • the aromatic ring e.g., phenyl, indole, thiophene
  • the aromatic ring can be substituted at one or more ring positions with such substituents as described above, as for example, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato, cyano, amino (including alkyl amino, dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino (including alkylcarbonylamino, aryl
  • Aryl groups can also be fused or bridged with alicyclic or heterocyclic rings which are not aromatic so as to form a polycycle such as tetralin.
  • alkenyl and alkynyl include unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but that contain at least one double or triple bond respectively and at least two adjacent carbon atoms.
  • an "optional single/double bond” is represented by a solid line together with a dashed line, and refers to a covalent linkage between two carbon atoms which can be either a single bond or a double bond of either E- or Z-configuration where appropriate.
  • lower alkyl means an alkyl group as defined above, but having from one to ten carbons, more preferably from one to six carbon atoms in its backbone structure.
  • lower alkenyl and “lower alkynyl” have similar chain lengths (and at least two carbon atoms).
  • Preferred alkyl groups are lower alkyls.
  • heterocyclyl or “heterocyclic group” refer to 3- to 10-membered ring structures, more preferably 4- to 7-membered rings, which ring structures include one or more heteroatoms, e.g., two, three, or four.
  • Heterocyclyl groups include pyrrolidine, oxolane, thiolane, piperidine, piperazine, morpholine, lactones, lactams such as azetidinones and pyrrolidinones, sultams, sultones, and the like.
  • the heterocyclic ring can be substituted at one or more positions with such substituents as described above, including halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato, cyano, amino (including alkyl amino, dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates, sulfonato, sulfamoyl,
  • polycyclyl or “polycyclic group” refer to two or more cyclic rings (e.g., cycloalkyls, cycloalkenyls, cycloalkynyls, aryls and/or heterocyclyls) where two or more carbons are common to two adjoining rings, e.g., the rings are "fused rings". Rings that are joined through non-adjacent atoms are termed "bridged" rings.
  • Each of the rings of the polycycle can be substituted with such substituents as described above, as for example, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato, cyano, amino (including alkyl amino, dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate, sulfates, sulfonate, sulfamoyl,
  • heteroatom as used herein means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, sulfur and phosphorus.
  • aryl aldehyde refers to a compound represented by the formula Ar-C(O)H, where Ar is an aryl moiety (as described above) and -C(O)H is a formyl or aldehydo group.
  • the aryl aldehyde is a (substituted or unsubstituted) benzaldehyde.
  • a variety of aryl aldehydes are commercially available, or can be prepared by routine procedures from commercially available precursors.
  • Procedures for the preparation of aryl aldehydes include the Vilsmeier-Haack reaction (See, e.g., Mz, Adv. Org. Chem. 9, pt. 1, 225-342 (1976)), the Gatterman reaction (Truce, Org. React. 9, 37-72 (1957)), the Gatterman-Koch reaction (Crounse, Org. React. 5, 290-300 (1949)), and the Reimer-Tiemann reaction (Wynberg and Meijer, Org. React. 28, 1-36 (1982)).
  • the structure of some of the compounds of this invention includes asymmetric carbon atoms. It is to be understood accordingly that the isomers arising from such asymmetry (e.g., all enantiomers and diastereomers) are included within the scope of this invention unless indicated otherwise. Such isomers can be obtained in substantially pure form by classical separation techniques and by stereochemically controlled synthesis. That is, unless otherwise stipulated, any chiral carbon center may be of either (R)- or ( ⁇ -stereochemistry. Furthermore, alkenes can include either the E- or Z-geometry, where appropriate. Additionally, one skilled in the art will appreciate that the chemical structures as drawn may represent a number of possible tautomers, and the present invention also includes those tautomers.
  • the invention provides methods for treating, preventing, or inhibiting seizure-associated states, including epilepsy.
  • the invention provides a method for treating, preventing, or inhibiting epileptogenesis in a subject.
  • the method includes administering to a subject in need thereof a sufficient amount of a compound of the present invention which modulates, e.g., upregulates or downregulates, a process in a pathway associated with epileptogenesis such that epileptogenesis is treated, prevented, or inhibited in the subject.
  • compounds for administration to a subject include pyrimidines such as substituted uracils which may be used directly to treat, prevent, or inhibit seizure-associated state, or may be converted in vivo to active ⁇ -amino anionic compounds, or may act as a combination thereof.
  • pyrimidines such as substituted uracils which may be used directly to treat, prevent, or inhibit seizure-associated state, or may be converted in vivo to active ⁇ -amino anionic compounds, or may act as a combination thereof.
  • the compound useful in the methods of the present invention can be represented by the formula
  • X is S or O
  • R 1 is selected from the group consisting of H, benzyl, isopropyl, p-NO 2 -benzyl, benzoyl, and -CH 2 SCH 3 ;
  • R 2 is selected from the group consisting of H, CH 3 , -CH 2 CH 2 -phenyl, n-butyl, ethyl, n-propyl, phenyl, benzyl, and
  • R 3 is selected from the group consisting of H, Br, I, CH 3 , -C ⁇ CC 4 H 9 , -CH 2 N(H)-phenyl, -CH 2 N(H)CH 2 -phenyl, -CH 2 -O-benzyl, an alkylsulfide, and an alkyluracil; and R 4 is selected from the group consisting of H, benzyl, isopropyl, p-NO 2 -benzyl, and benzoyl; or pharmaceutically acceptable salts or esters thereof.
  • the compounds for use in accordance with the invention may be selected from compounds 1-80, discussed herein, and specifically listed in the Exemplification Section.
  • many uracil analogs described herein have demonstrated broad spectrum anti-convulsant activity (active on primary or secondary seizure types), and virtually no neurotoxicity in various biological tests, outperforming established anticonvulsant drugs.
  • Another embodiment of the invention relates to a method of treating, preventing, or inhibiting a seizure-associated state in a subject comprising administering to a subject in need thereof a sufficient amount of a compound of the present invention, such that the seizure-associated state is treated, prevented, or inhibited.
  • the invention is a method of treating, preventing, or inhibiting ictogenesis in a subject comprising administering to a subject in need thereof a sufficient amount of a compound of the present invention, such that ictogenesis is treated, prevented, or inhibited.
  • the invention is a method of treating, preventing, or inhibiting epileptogenesis in a subject comprising administering to a subject a sufficient amount of a compound of the present invention, such that epileptogenesis is treated, prevented, or inhibited.
  • the step of administering to the subject can include administering to the subject a compound which is metabolized to an anti-convulsant and/or anti-epileptogenic compound of the invention.
  • the methods of the invention include the use of prodrugs which are converted in vivo to the therapeutic compounds of the invention. See, e.g., Silverman, ch. 8, cited above. Such prodrugs can be used to alter the biodistribution to allow compounds which would not typically cross the blood-brain barrier to cross the blood-brain barrier, or the pharmacokinetics of the therapeutic compound.
  • an anionic group e.g., a carboxylate group
  • an anionic group can be esterified with an ethyl or a fatty group to yield a carboxylic ester.
  • the ester can be cleaved, enzymatically or non- enzymatically, to reveal the anionic group.
  • the methods of the invention include administering to the subject a derivative of uracil or an analog thereof (including substituted pyrimidines, uridine 5'-monophosphate (UMP) and uridine, or analogs thereof).
  • Administration of a uracil compound or metabolite thereof can result in the in vivo formation of an active compound of the invention.
  • the methods of the invention may include the step of administering to a subject in need thereof a sufficient amount of a substituted or unsubstituted uracil, or a derivative or analog thereof (or a pharmaceutically acceptable salt or ester thereof), in an amount effective to treat, prevent, or inhibit a seizure-associated state , e.g., by direct activity of the uracil, in vivo conversion of the uracil to a ⁇ -amino acid compound, or by a combination thereof.
  • the compound of the invention is not uridine.
  • Glutamate Glutamate
  • Aspartate Aspartate
  • GABA Glycine [26-30]
  • Taurine GABA
  • NMDA N-methyl-D- aspartate
  • GABA gamma-amino-butyric acid
  • an agent to be administered to a subject to treat, prevent, or inhibit epileptogenesis may treat, prevent, or inhibit one or more processes in at least one pathway associated with epileptogenesis.
  • an agent useful for inhibition of epileptogenesis can reduce the release of, or attenuate the epileptogenic effect of, NO in brain tissue; antagonize an NMDA receptor; augment endogenous GABA inhibition; block voltage-gated ion channels; reduce the release of, reduce the free concentration of (e.g., by chelation), or otherwise reduce the epileptogenic effect of cations including Ca 2+ , Zn 2+ , or Fe 2+ ; inhibit oxidative cell damage; or the like.
  • an agent.to be administered to a subject to inhibit epileptogenesis is capable of inhibiting at least two processes in at least one pathway associated with epileptogenesis.
  • Non-limiting examples of pharmacophores which can modulate a process in a pathway associated with epileptogenesis include:
  • ⁇ inhibitors of NO synthase such as L-arginine and alkylated derivatives thereof;
  • ⁇ antagonists of NMDA receptors such as (i?)- ⁇ -amino acids. See, e.g., Leeson, P.D. and Iverson, L.L., J Med. Chem. (1994) 37:4053-4067, for a general review of inhibitors of the NMDA receptor;
  • ⁇ augmenters of endogenous GABA inhibition such as inactivators of GABA aminotransferase like gamma-vinyl-GABA. See, e.g., Krogsgaard-Larsen, P., et al., J. Med. Chem. (1994) 37:2489-2505) for a review of GABA receptor agonists and antagonists;
  • ⁇ chelators of Ca 2+ , Zn 2+ , or Fe 2+ such as EDTA, EGTA, TNTA, 2,2- bipyridine-4,4,-dicarboxylate, enterobactin, porphyrins, crown ethers, and azacrown ethers; and
  • the agent antagonizes an NMDA receptor and augments endogenous GABA inhibition, hi certain particular embodiments, the agent is administered orally.
  • the agent is transported to the nervous system of the subject by an active transport shuttle mechanism.
  • an active transport shuttle is the large neutral amino acid transporter, which is capable of transporting amino acids across the blood-brain barrier (BBB).
  • Voltage-gated ion channels can play a significant role in neuronal functions.
  • phenytoin binds Na + channels, and upon firing, the channels open, which in turn results in better binding to the channels by phenytoin, i.e., selective state-dependent binding.
  • phenytoin binds Na + channels, and upon firing, the channels open, which in turn results in better binding to the channels by phenytoin, i.e., selective state-dependent binding.
  • phenytoin binds Na + channels, and upon firing, the channels open, which in turn results in better binding to the channels by phenytoin, i.e., selective state-dependent binding.
  • phenytoin binds Na + channels, and upon firing, the channels open, which in turn results in better binding to the channels by phenytoin, i.e., selective state-dependent binding.
  • these drugs appear to be effective against the high frequency repetitive firing required for the generation and progression of an epileptic seizure without significant interference with the normal firing modes.
  • such drugs are active in the MES-induced seizure
  • voltage-gated ion channels that are targeted in antiseizure drug development include those affecting calcium and potassium ions.
  • phenytoin, benzodiazepines, and barbiturates are known to affect K + currents or the Ca 2+ dependent K + current.
  • calcium ion entry into neurons occurs through several heterogeneous types of voltage-dependent calcium ion channels composed of multiple subunits.
  • the anticonvulsant drug ethosuximide is known to selectively reduce low threshold T-type Ca 2+ channel currents in a voltage-dependent manner at therapeutic drug concentration [56].
  • This anticonvulsant is effective in absence seizure but ineffective in focal seizures or suppressing the development of primary generalized tonic-clonic seizures (as shown in the table below).
  • absence seizures may also be suppressed by benzodiazepines through the enhancement of GABA receptors .
  • the main inhibitory neurotransmitters in the CNS are glycine and GABA with the latter being the most predominant and effective in the brain inhibition processes.
  • Glycine is an agonist at two neurotransmitter receptor sites, one sensitive to inhibition by strychnine and linked to the inhibitory glycine-gated chloride channel and the other a co- agonist site on the excitatory amino acid NMDA cation channel complex [61, 62].
  • all neurons within the mammalian CNS, including the human brain have GABA receptors that produce hyperpolarizing inhibition when activated.
  • reduction of GABA-mediated inhibition results in seizures while enhancement of inhibition is associated with the suppression of seizures [63].
  • GABA A receptor appears to be more important in controlling neuronal hyperexcitability, and thus seizure propagation.
  • GABA glutamic acid (or glutamate) decarboxylase
  • GABA A receptor binds to a GABA A receptor, and mediates inhibition by opening the chloride ion channels and hyperpolarizing the membrane.
  • GABA uptake carrier After synaptic release, GABA is taken up into nerve cells or glial cells through a GABA uptake carrier. Subsequently, in the neuron, GABA is either re-released or broken down by the GABA transaminase (GABA-T) into succinic acid semialdehyde (SSA) as in glial cells.
  • GABA-T GABA transaminase
  • SSA succinic acid semialdehyde
  • the GABA A receptor is associated with an ion channel permeable to chloride ions, GABA increases the probability of the ion channel opening that increases the inward flow of chloride ions through the nerve cell membrane and inhibits neuronal firing.
  • Drugs such as the barbiturates and benzodiazepines exert their anticonvulsant activity by modifying GABA receptor mediated responses via associated modulatory sites to potentiate the effect on chloride ion conductance [64-66].
  • anticonvulsant drugs that exert their effects by enhancing the actions of GABA at GABAA receptors.
  • Barbiturates such as phenobarbital act to prolong inhibition by keeping GAB A-activated channels open longer; and benzodiazepines such as diazepam, clonazepam and clobazam act to increase the frequency of chloride ion channel opening, thereby making inhibitory postsynaptic potentials larger [67].
  • These two classes of drugs bind to specific sites on the GABA receptor and allosterically alter the channel properties.
  • Other approaches to enhance GABA mediated inhibition include, but are not limited to:
  • GABA mimetics since GABA is not sufficiently lipophilic to effectively cross the blood brain-barrier when systematically administered;
  • S GABA-T enzyme inhibitors preventing the breakdown of GABA; and S GABA uptake blockers in neurons or glial cells [68].
  • Inhibition enhancement may also be achieved by interfering with the GABA catabolic enzyme GABA-T. This may increase the synaptic availability of GABA as its breakdown to SSA is decreased or prevented.
  • GABA-T inhibitors should potentiate the action of synaptically released GABA without producing generalized inhibitory effects throughout the brain, as do direct-acting GABA agonists. This approach led to the development and subsequent marketing of the anticonvulsant drug, vigabatrin, an irreversible GABA-T inhibitor [70].
  • GABA uptake blockers should be expected to selectively potentiate the inhibitory effects of synaptically released GABA [71].
  • Conventional blockers of GABA uptake such as nipecotic acid and guavacine, have anticonvulsant activity when injected intracerebroventricularly, but they do not cross the BBB [72]. Thus it increases the extracellular concentration of GABA in the hippocampus and prolongs the inhibitory effects of synaptically released GABA.
  • GABA uptake is performed by carriers or transporters, which exist in four different types known as the GAT-I, GAT-2, GAT-3 and GAT-4.
  • GAT-I which is found in both neurons and glial cells, and is readily inhibited by nipecotic acid or its analogue Tiagabine, and exclusively transports GABA [75].
  • GAT-2 and GAT-3 which have a glial localization, bind both GABA and ⁇ -alanine.
  • GAT-4 may be absent from the CNS.
  • Aspartate and Glutamate are the main excitatory neurotransmitters in the brain, the latter being more predominant.
  • Glutamate produces an excitatory postsynaptic current that may be resolved into fast and slow (or long- lasting) components.
  • the latter being mediated through NMDA receptors while the former is controlled by the non-NMDA ionotropic glutamate receptor-channels AMPA and Kainate [79, 80].
  • Glutamate acts postsynaptically on three classes of ion channel-linked receptors named after their preferred agonists: N-Methyl-D-aspartic Acid (NMDA), kainic acid and alpha-Amino-3-hydroxy-5-Methyl-4-isoxazole Propionic Acid (AMPA) or quisqualic acid.
  • NMDA N-Methyl-D-aspartic Acid
  • AMPA alpha-Amino-3-hydroxy-5-Methyl-4-isoxazole Propionic Acid
  • quisqualic acid quisqualic acid.
  • the latter two molecules, AMPA and quisqualic acid share the same receptor, which is usually referred to as the AMPA receptor.
  • the NMDA receptor has been shown to exhibit properties that facilitate the burst firing and Paroxysmal Depolarizing Shift (PDS) typical of epileptic discharges [82]. Moreover, excessive activation of NMDA receptors is thought to mediate Ca 2+ dependent neurotoxicity associated with trauma, epilepsy and several neurodegenerative diseases.
  • PDS Paroxysmal Depolarizing Shift
  • NMDA receptors are present in virtually every neuronal cell [74]. They are widely distributed in the brain and spinal cord, with the highest densities in cerebral cortex and hippocampus, areas of the brain that seem to be the most susceptible to epilepsy pathogenesis. These receptors, when activated, control ion channel permeability to Na + , K + , and Ca 2+ . However, the channel is blocked by Mg 2+ during normal synaptic transmission and is voltage-dependent. For activation, NMDA receptors require occupation of both the glutamate and glycine subunits. Thus, NMDA receptors can be blocked by competitive antagonists acting at either the glutamate or the glycine (co-agonist) sites.
  • the NMDA receptor is associated with several modulators and pharmacological antagonists which act at its different sites. These include the NMDA (or glutamate) site, the glycine (co-agonist) site, the Zn 2+ modulatory site, the redox modulatory site, the polyamine site, the Mg 2+ -block site, the H + ion modulation sites as well as other sites involved in the binding of open-channel blockers such as memantine and the dissociative anesthetic agents phencyclidine (l-(l-PhenylCyclohexyl)Piperidine (PCP) and dizocilpine, also known as MK-801.
  • the invention provides a method for treating, preventing, or inhibiting a seizure-associated state in a subject.
  • the method includes the step of administering to a subject in need thereof a sufficient amount of an agent which a) blocks sodium or calcium ion channels, or opens potassium or chloride ion channels; and/or b) has at least one activity selected from the group consisting of NMDA receptor antagonism; augmentation of endogenous GABA inhibition; calcium binding; iron binding; zinc binding; NO synthase inhibition; and antioxidant activity; such that seizure-associated state is treated, prevented, or inhibited in the subject.
  • an active agent of the invention antagonizes NMDA receptors, e.g., by binding to the glycine binding site of the NMDA receptors.
  • the agent augments GABA inhibition by decreasing glial GABA uptake.
  • the agent is administered orally.
  • the method further includes administering the agent in a pharmaceutically acceptable vehicle.
  • the invention provides a method for treating, preventing, or inhibiting both ictogenesis and epileptogenesis in a subject, i.e., concurrently or sequentially.
  • the method includes the step of administering to a subject in need thereof a sufficient amount of an agent which a) blocks sodium or calcium ion channels, or opens potassium or chloride ion channels; and b) has at least one activity selected from the group consisting of NMDA receptor antagonism; augmentation of endogenous GABA inhibition; calcium binding; iron binding; zinc binding; NO synthase inhibition; and antioxidant activity; such that epileptogenesis is treated, prevented, or inhibited in the subject.
  • one or more moieties or pharmacophores that are associated with the modulation the pathways described herein may be coadministered with the compounds of the invention, either as substituents of (i.e., covalently linked to) the compounds of the invention or in combination treatments as separate agents.
  • one or more existing drugs may be used in conjunction with the compounds of the present invention to achieve a therapeutic effect, wherein the existing drug may be a substituent of the compounds of the present invention or an additional agent coadministered with the compound of the invention.
  • blockers of sodium and/or calcium ion channel activity are well known in the art and can be used as a substituent of the compounds in the methods of the present invention.
  • any compound which opens potassium or chloride ion channels can be used as a substituent.
  • Antagonists of NMDA receptors and augmenters of endogenous GABA inhibition are also known to one of skill in the art and can be used used as a substituent.
  • 2,3-quinoxalinediones are reported to have NMDA receptor antagonistic activity (See, e.g., U.S. Patent No.
  • Exemplary calcium and zinc chelators include moieties known in the art for chelation of divalent cations, including ethylenediaminetetraacetic acid (EDTA), ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid, and the like, in addition to those mentioned supra.
  • EDTA ethylenediaminetetraacetic acid
  • ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid and the like, in addition to those mentioned supra.
  • Exemplary iron chelators include enterobactin, pyridoxal isonicotinyl hydrazones, N 3 N'- bis(2-hydroxybenzoyl)-ethylenediamine-N,N'-diacetic acid (HBED), and 1-substituted- 2-alkyl-3-hydroxy-4-pyridones, including 1 -(2'-carboxyethyl)-2-methyl-3-hydroxy-4- pyridone, and other moieties known in the art to chelate iron.
  • enterobactin enterobactin
  • pyridoxal isonicotinyl hydrazones N 3 N'- bis(2-hydroxybenzoyl)-ethylenediamine-N,N'-diacetic acid (HBED)
  • HBED N 3 N'- bis(2-hydroxybenzoyl)-ethylenediamine-N,N'-diacetic acid
  • N ⁇ -substituted arginine analogs especially of the L-configuration, including L-N ⁇ -nitro-arginine (a specific inhibitor of cerebral NO synthase), L-N ⁇ -amino-arginine, and L-N ⁇ -alkyl-arginines; or an ester thereof, preferably the methyl ester.
  • exemplary antioxidants include ascorbic acid, tocopherols including alpha-tocopherol, and the like.
  • a linkage between, for example, a compound of the invention and a pharmacophore can be selected such that the two pharmacophores are cleaved from each other in vivo (e.g. , by the selection of a linkage which is labile in vivo).
  • a linkage which is labile in vivo examples of such biologically labile linkages are known in the art. See, e.g., Silverman, cited above.
  • a "hybrid" drug can be designed to be transported within the body to reach a site or organ such as the brain, where one or more of the moieties exert a biological effect, at which site the hybrid drug can be cleaved to provide two active drug moieties.
  • the invention provides a method for concurrently treating, preventing, or inhibiting epileptogenesis and ictogenesis, the method including the step of administering to a subject in need thereof a sufficient amount of a compound of the formula:
  • X is S or O
  • R 1 is selected from the group consisting of H, benzyl, isopropyl, p-NO 2 -benzyl, benzoyl, and -CH 2 SCH 3 ;
  • R 2 is selected from the group consisting of H, CH 3 , -CH 2 CH 2 -phenyl, n-butyl, ethyl, n-propyl, phenyl, benzyl, and -CH 2 CH(OH)-phenyl;
  • R 3 is selected from the group consisting of H, Br, I, CH 3 , -C ⁇ CC 4 H 9 , -CH 2 N(H)-phenyl, -CH 2 N(H)CH 2 -phenyl, -CH 2 -O-benzyl, an alkylsulfide, and an alkyluracil; and R 4 is selected from the group consisting of H, benzyl, isopropyl, p-NO 2 -benzyl, and benzoyl; or pharmaceutically acceptable salts or esters thereof.
  • the invention provides a method for treating, preventing, or inhibiting a disorder associated with NMDA receptor activity. The method includes the step of administering to a subject in need thereof a sufficient amount of a compound of the formula:
  • X is S or O
  • R 1 is selected from the group consisting of H, benzyl, isopropyl, p-NO 2 -benzyl, benzoyl, and -CH 2 SCH 3 ;
  • R 2 is selected from the group consisting of H, CH 3 ,
  • R 3 is selected from the group consisting of H, Br, I, CH 3 , -C ⁇ CC 4 H 9 , -CH 2 N(H)-phenyl, -CH 2 N(H)CH 2 -phenyl, -CH 2 -O-benzyl, an alkylsulfide, and an alkyluracil; and
  • R 4 is selected from the group consisting of H, benzyl, isopropyl, p-NO 2 -benzyl, and benzoyl; or pharmaceutically acceptable salts or esters thereof.
  • Compounds which find use in the therapeutic methods of the invention can be determined through routine screening assays.
  • the animal model of Phase 1 epileptogenesis described in the Exemplification, infra can be employed to determine whether a particular compound has anti-epileptogenic activity against Phase 1 epileptogenesis.
  • Chronic epileptogenesis can be modeled in rats (and candidate compounds screened) with the kindling assay described by Silver et al. ⁇ Ann. Neurol. (1991) 29:356).
  • compounds useful as anticonvulsants can be screened in conventional animal models, such as the mouse model described in Horton, R. W. et al. , Eur. J. Pharmacol. (1979) 59:75-83.
  • Binding to the glycine site on an NMDA receptor can be quantified, e.g., according to the method described in Kemp, A., et al, Proc. Natl. Acad. ScI USA (1988) 85:6547. Effect on the voltage-gated Na + channel can be evaluated in vitro by voltage clamp assay in rat hippocampal slices.
  • Another embodiment of the invention pertains to a method of treating, preventing, or inhibiting seizures in a subject suffering from head trauma comprising administering to a subject a sufficient amount of a compound of the present invention, such that seizures are treated, prevented, or inhibited.
  • the seizures are caused by epileptogenesis.
  • Another embodiment of the invention pertains to a method of treating, preventing, or inhibiting epileptogenesis in a subject suffering from head trauma comprising administering to a subject a sufficient amount of a compound of the present invention, such that epileptogenesis is treated, prevented, or inhibited.
  • Another embodiment of the invention pertains to a method of treating, preventing, or inhibiting seizures in a subject suffering from traumatic brain injury (TBI) comprising administering to a subject a sufficient amount of a compound of the present invention, such that seizures are treated, prevented, or inhibited, e.g., wherein the seizures are caused by epileptogenesis.
  • TBI traumatic brain injury
  • Another embodiment of the invention pertains to a method of treating, preventing, or inhibiting epileptogenesis in a subject suffering from traumatic brain injury (TBI) comprising administering to a subject a sufficient amount of a compound of the present invention, such that epileptogenesis is treated, prevented, or inhibited.
  • TBI traumatic brain injury
  • Neurons that are the most prone to epilepsy are primarily located in the cerebral cortex, which includes the frontal, parietal, occipital and temporal lobes, with the latter lobe appearing to be the most predisposed region of the brain to the process of epileptogenesis.
  • a seizure focus may therefore be defined as that part of the brain from which seizures arise and eventually spread.
  • the anatomical location of this focus inside the brain may determine the observed symptoms or the seizure type.
  • the epileptic seizures may be primarily characterized by motor shaking of the body; while difficulties with speech are usually the first signs of seizures with seizure foci located in the temporal lobe region.
  • Any infectious or physical trauma to the CNS is a potential cause for epilepsy.
  • Prenatal trauma, congenital defects, birth trauma, e.g., birth anoxia, cerebral palsy, and degenerative neurological diseases or postnatal insults, such as brain trauma, CNS infections, brain tumors, and cerebrovascular diseases greatly increase the incidence of epilepsy.
  • metabolic or genetic abnormalities may also play a significant role in the predisposition to developing this disorder.
  • KCC2 mouse gene encoding the neuronal-specif ⁇ c K-Cl cotransporter, which is thought to participate in the postnatal brain development of inhibitory mechanisms by driving intracellular chloride ions to low concentrations and shirting the reversal potential for chloride ion conductances, was associated with hyperexcitability and epilepsy.
  • animals deficient in KCC2 exhibited frequent generalized seizures and died shortly after birth suggesting that these genes play a key role in controlling CNS excitability at birth as well as in adult life.
  • Epileptic seizures are very complex and their clinical manifestations may include an array of sensory, motor and autonomic phenomena.
  • the Commission on Classification and Terminology of the International League against Epilepsy ILAE
  • ILAE International League against Epilepsy
  • EEG electroencephalographic
  • the ILAE seizure classification system distinguishes three main categories of seizures, which are further divided in subcategories, and attempts to describe seizures with respect to six major aspects including, for example, clinical seizure type, electroencephalographic (EEG) seizure type, i.e. the EEG patterns during a clinical seizure, EEG interictal expression, i.e. EEG patterns between clinical seizures, anatomical substrate, etiology, and age.
  • EEG electroencephalographic
  • Partial seizures also known as focal or local seizures, are the most common seizures occurring in about two-thirds of the epilepsy patient population.
  • the first clinical and EEG changes indicate an initial activation of a system of neurons limited to one cerebral hemisphere.
  • consciousness i.e. degree of awareness or responsiveness of the patient to externally applied stimuli
  • the seizure is classified as a complex partial seizure; otherwise, it is a simple partial seizure.
  • simple partial seizures may evolve to complex partial seizures and/or generalized tonic-clonic convulsions, also called secondary generalized seizures.
  • Epilepsy patients suffering from complex partial seizures usually experience an impairment of consciousness and psychic symptoms that proceed from disturbance of higher cerebral functions.
  • Simple partial seizures may involve motor signs of any portion of the body depending on the site of origin of the seizure attack in the motor strip in the cortex. These seizures may remain strictly focal or they may spread to contiguous cortical areas to produce a sequential involvement of body parts in an epileptic "march" leading to a seizure type known as the Jacksonian seizure.
  • Simple partial seizures may also involve autonomic signs (vomiting, flushing, sweating, piloerection, pupil dilation) or somatosensory symptoms, which arise from the cortical areas subserving sensory functions.
  • patients may experience vertiginous symptoms (sensations of falling in space or floating); visual hallucinatory phenomena involving people or scenes; auditory sensations like music; olfactory sensations, usually in the form of unpleasant odors; gustatory sensations that may be odious or pleasant taste hallucinations varying in elaboration from crude (sweet, bitter, salty, or sour) to sophisticated.
  • Generalized seizures are primary generalized seizures that occur in about one- third of the epilepsy patient population [15]. They may be further divided into three major subcategories including, for example, absence seizures, myoclonic seizures and generalized tonic clonic seizures.
  • the first clinical signs of generalized seizures indicate an involvement of both cerebral hemispheres. Their initial manifestations may include the impairment of consciousness, bilateral movements as well as bilateral ictal EEG patterns that reflect the extent of neuronal discharges in the two hemispheres. Absence seizures
  • Abesence seizures also known as "petit mal" are non-convulsive seizures that occur in childhood and usually tend to cease in late teenage years.
  • the hallmark of their attack is a sudden onset, interruption of ongoing activities, a blank stare, possibly a brief upward rotation of eyes.
  • the attack is usually brief, lasting about 30 seconds or less, and dissipating as quickly as it started.
  • LGS Lennox-Gastaut Syndrome
  • consciousness impairment In circumstances wherein consciousness impairment is the only clinical sign during the seizure attack, it is a simple absence; otherwise, it may either be the Lennox- Gastaut Syndrome or the complex absence.
  • the EEG of complex absences is characterized by three per second spikes and slow discharge during ictal periods.
  • the seizures are extremely brief and frequently incorporate many symptoms of simple and complex absences.
  • the EEG for LGS is diffusely slow with typical spikes and wave discharges at frequencies other than 3 Hertz. Although rare, absence seizures may sometimes evolve into generalized tonic-clonic activity producing a seizure-type that is different from the complex partial seizures with secondary generalization.
  • absence seizures include, for example, absence seizures with impairment of consciousness only (Simple absence), wherein no other activities, beside consciousness impairment, take place during the attack; absence seizures with mild clonic components, wherein the onset is indistinguishable from the above but clonic movements may occur in the eyelids, at the corner of the mouth or in other muscle groups with severity varying from almost imperceptible movements to generalized myoclonic jerks; absence seizures with atonic components, which is characterized by a diminution in tone of muscles sub-serving posture as well as limbs leading to drooping of the head, occasional slumping of the trunk, dropping of the arms, and relaxation of the grip (however, it is rare for the muscle tone to be so diminished that the patient falls down); absence seizures with tonic components, wherein tonic muscular contractions may occur during the seizure attack leading to increase in muscle tone that may affect the extensor or flexor muscles, and the head may tonically draw to one side or the other (if the patient is standing, the head
  • Myoclonic seizures are characterized by single or multiple quick involuntary jerks known as myoclonus or myoclonic jerks. These jerks are sudden, brief shock-like contractions, which may be generalized or confined to the face and trunk or to one or more extremities or even to individual muscles or groups of muscles. Myoclonic seizure attacks occur predominantly around the hours of going to sleep or awakening from sleep.
  • Generalized tonic clonic seizures or "grand mal” seizures occur in both childhood and adult life but they are not as frequent as absence seizures. Their frequency may vary from one a day, to one every three months, or even to one every few years. "Grand mal” is the most common type of generalized seizure. Although the majority of patients lose consciousness without any premonitory symptoms, some may experience a vague ill-described warning. During the attack, there is a sudden sharp tonic contraction of muscles and the patient falls to the ground in the tonic state, passing urine involuntarily and injuring his/her tongue, if it is caught between the teeth, as the jaw contracts. The tonic contraction inhibits respiration, possibly, leading to cyanosis (discoloration of the skin due to lack of oxygen in the blood).
  • the clonic phase is characterized by convulsive movements and small gusts of grunting respiration within movement intervals. As the seizure approaches its end, the frequency of clonic contractions gradually decreases and saliva may froth from the mouth. Finally, deep respiration occurs and all the muscles relax but the patient remains unconscious for several minutes, and then awakes feeling stiff and diffusely sore.
  • Unclassified epileptic seizures This category includes all seizures that cannot be classified in either of the two previously described categories due to inadequate or incomplete data. It includes febrile seizures, status epilepticus, and neonatal seizures. Febrile seizures, usually, occur during the rising phase of a fever in children (age: six months to five years). Status epilepticus refers to repetitive seizures lasting more than 30 minutes without total recovery between attacks. This is most frequently observed in generalized seizure patients and constitutes a medical emergency that may result in brain damage. Many of the seizures occurring in infants and newborns including rhythmic eye movements, chewing, swimming movements, jittering and apnea are classified in this category.
  • the invention provides compounds useful for the treatment, prevention, or inhibition of seizure-associated states, including epileptogenesis and ictogenesis.
  • Molecules such as ⁇ -alanines that have neuroprotective properties may prevent primary and/or secondary epileptogenesis.
  • such molecules may prevent a focus from forming or prevent an existing focus from enlarging and worsening (e.g., increasing in size or increasing in number) respectively.
  • they may be considered as not only anticonvulsants but also anti-epileptogenic drugs.
  • Drugs developed for the treatment or management of epilepsy will be required to fulfill certain requirements in order to exert a therapeutic biological activity inside the brain. Due to the properties of the BBB, ⁇ -alanine and other charged molecules are difficult candidates for drug development in the therapy of neurological disorders.
  • the prodrug approach may allow us to circumvent this problem using uracil analogs as they are metabolized in vivo to ⁇ -amino acids following the path illustrated below.
  • the invention is directed to prodrugs that are active to treat, prevent or inhibit seizure-associated states, e.g., epileptogenesis or ictogenesis, prior to metabolization.
  • Hydantoins are prototypic anticonvulsants, which bind Na+ channels.
  • the structure of the hydantoin moiety is depicted below
  • the compounds of the present invention may also have anti-convulsant activity.
  • uracil-based drugs may therefore be expected to exhibit dual biological activity; (1) as anticonvulsants mediating their biological effects via the voltage-gated sodium ion channels due to their similarity with hydantoins, and (2) as antiepileptogenic prodrugs that are anticipated to metabolize to ⁇ -alanine- based drugs.
  • the enzymatic catabolism of uracils as illustrated below indicates that the C-4, C-5, C-6 and N-I atoms of the uracil ring are preserved from the parent uracil to the corresponding ⁇ -amino acid. dihydrouracil dehydrogenase
  • substituted or unsubstituted uracils, and derivatives or analogs thereof may be especially advantageous as certain uracil compounds have been found to have anti-ictogenic properties when tested in an anti-seizure model in rats. See, e.g.,
  • the prodrug form of the compound i.e., a uracil
  • the metabolically-produced ⁇ -amino anionic compounds can have anti-epileptogenic and/or anti-ictogenic activity.
  • pyrimidine compounds such as 5- fluorouracil (5FU)
  • 5FU 5- fluorouracil
  • the anti-cancer activity of 5FU and similar compounds is believed to be due to a "suicide substrate" mechanism where the 5FU inhibits thymidylate synthase, an enzyme important in DNA synthesis.
  • pyrimidine compounds administered according to the invention for the treatment, prevention, or inhibition of seizure- associated states do not significantly inhibit thymidylate synthase.
  • inhibition of thymidylate synthase by pyrimidine compounds is increased by the presence of electronegative groups at the 5 -position of the pyrimidine ring, and can therefore be decreased by providing such compounds with non-electronegative groups at the 5-position of the pyrimidine ring. It is further believed that by providing substituents with sufficient steric bulk to decrease the ability of the pyrimidine compound to bind to thymidylate synthase, inhibition of thymidylate synthase can be decreased.
  • the compounds of the present invention can be represented by the formula
  • X is S or O
  • R 1 is selected from the group consisting of H, benzyl, isopropyl, p-NO 2 -benzyl, benzoyl, and -CH 2 SCH 3 ;
  • R 2 is selected from the group consisting of H, CH 3 , -CH 2 CH 2 -phenyl, n-butyl, ethyl, n-propyl, phenyl, benzyl, and
  • R 3 is selected from the group consisting of H, Br, I, CH 3 ,
  • R 4 is selected from the group consisting of H, benzyl, isopropyl, p-NO 2 -benzyl, and benzoyl; or acceptable salts, e.g., pharmaceutically acceptable salts, or esters thereof.
  • at least one of R , R , R , or R is not hydrogen, e.g., at least two of R 1 , R 2 , R 3 , or R 4 are not hydrogen, e.g., at least three of R 1 , R 2 , R 3 , or R 4 are not hydrogen, e.g. , wherein R 1 , R 2 , R 3 , and R 4 are not hydrogen.
  • R 4 is not hydrogen.
  • the compound is selected from the group consisting of H, benzyl, isopropyl, p-NO 2 -benzyl, and benzoyl; or acceptable salts, e.g., pharmaceutically acceptable salts, or esters thereof.
  • R 1 is selected from the group consisting of H, benzyl, isopropyl, p-NO 2 -benzyl, benzoyl, and -CH 2 SCH 3 ;
  • R 2 is selected from the group consisting of H, CH 3 , -CH 2 CH 2 -phenyl, n-butyl, ethyl, n-propyl, phenyl, benzyl, and -CH 2 CH(OH)-phenyl;
  • R 3 is selected from the group consisting of H, Br, I, CH 3 , -C ⁇ CC 4 H 9 , -CH 2 N(H)-phenyl, -CH 2 N(H)CH 2 -phenyl,
  • R 4 is selected from the group consisting of H, benzyl, isopropyl, p-NO 2 -benzyl, and benzoyl; or acceptable salts, e.g., pharmaceutically acceptable salts, or esters thereof.
  • R 1 , R 2 , and R 4 are not H, while R 3 is H, Br, I, -C ⁇ CC 4 H 9 , or CH 3 .
  • R 1 and R 4 are not H, while R 2 is CH 3 , and R 3 is H or Br.
  • R 1 and R 4 are not H, while R 2 and R 3 are CH 3 . In certain embodiments, R and R are not H, while R 3 is Br, and R 1 is benzyl or isopropyl. In certain embodiments, R 3 and R 4 are not H, while R 2 is H or CH 3 , and R 1 is benzyl. In certain embodiments, R 2 is not H, while R 1 is benzyl, R 3 is -C ⁇ CC 4 H 9 , and R 4 is H or benzyl. In certain embodiments, R 2 is not H, while R 1 and R 4 is p-NO 2 -benzyl, and R 3 is Br. In certain embodiments, R 2 is not CH 3 , while R 1 and R 4 is p-NO 2 -benzyl, and R 3 is H. In another particular embodiment, the compound is
  • R 1 is selected from the group consisting of H, benzyl, p-NO 2 -benzyl, and benzoyl;
  • R 2 is selected from the group consisting of H, CH 3 , -CH 2 CH 2 -phenyl, and n-butyl;
  • R 3 is selected from the group consisting of H, Br, I, and -C ⁇ CC 4 H 9 ; and R 4 is selected from the group consisting of H, benzyl, p-NO 2 -benzyl, and benzoyl; or acceptable salts, e.g., pharmaceutically acceptable salts, or esters thereof.
  • R 1 , R 2 , and R 4 are not H, while R 3 is H, Br, I, or
  • R 1 and R 4 are not H, while R 2 is CH 3 , and R 3 is H or Br. In certain embodiments, R 2 and R 4 are not H, while R 3 is Br, and R 1 is benzyl. In certain embodiments, R 3 and R 4 are not H, while R 2 is H or CH 3 , and R 1 is benzyl. In certain embodiments, R 2 is not H, while R 1 is benzyl, R 3 is -C ⁇ CQHg, and R 4 is H or benzyl. In certain embodiments, R 2 is not H, while R 1 and R 4 is p-NO 2 -benzyl, and R 3 is Br. hi certain embodiments, R 2 is not CH 3 , while R 1 and R is p-NO 2 -benzyl, and R 3 is H.
  • R 2 is selected from the group consisting of H, phenyl, benzyl, and
  • R 3 is selected from the group consisting of H, Br, -CH 2 N(H)-phenyl, and
  • R 2 is not H, while R 3 is H or Br. In certain embodiments, R 2 is not H, while R 3 is Br. In yet another particular embodiment, the compound is
  • R 1 is selected from the group consisting of H, benzyl, isopropyl, p-NO 2 -benzyl, benzoyl, and -CH 2 SCH 3 ;
  • R 2 is selected from the group consisting of H, CH 3 ,
  • R 3 is selected from the group consisting of H, Br, I, CH 3 ,-C ⁇ CC 4 H 9 , -CH 2 N(H)-phenyl, -CH 2 N(H)CH 2 -phenyl, -CH 2 -O-benzyl, an alkylsulfide, and an alkyluracil; and
  • R 4 is selected from the group consisting of H, benzyl, isopropyl, p-NO 2 -benzyl, and benzoyl; or acceptable salts, e.g., pharmaceutically acceptable salts, or esters thereof.
  • the compound is
  • R 1 is selected from the group consisting of H and -CH 2 SCH 3 ; and R 2 is selected from the group consisting of H, n-butyl, ethyl, n-propyl, phenyl, and benzyl; or acceptable salts, e.g., pharmaceutically acceptable salts, or esters thereof.
  • the compound is
  • n 0 or 1 ;
  • R 5 is O or absent
  • R 6 is selected from the group consisting of CH 3 and benzyl
  • R 7 is selected from the group consisting of H and phenyl; or acceptable salts, e.g., pharmaceutically acceptable salts, or esters thereof. In certain embodiments, R 7 is not H, while R 6 is CH 3 and R 5 is absent. In yet another particular embodiment, the compound is
  • R 1 and R la are independently selected from the group consisting of H, benzyl, p-NO 2 -benzyl, benzoyl, and -CH 2 SCH 3 ;
  • R 2 and R 2a are independently selected from the group consisting of H, CH 3 ,
  • R and R a are independently selected from the group consisting of H, benzyl, p-NO 2 -benzyl, and benzoyl;
  • R 7 is selected from the group consisting of H and phenyl; or acceptable salts, e.g., pharmaceutically acceptable salts, or esters thereof, hi certain embodiments, R 1 , R la , R 4 and R 4a are not all benzyl. In yet another particular embodiment, the compound is
  • R 1 and R la are independently selected from the group consisting of H and benzyl
  • R 2 and R 2a are independently selected from the group consisting of H, CH 3 , and benzyl;
  • R 7 is selected from the group consisting of H and phenyl; or acceptable salts, e.g., pharmaceutically acceptable salts, or esters thereof.
  • the compound is
  • the compound of the invention is not a compound of published U.S. Utility Application, publication number 20030153584-A1, the contents of which are hereby expressly incorporated herein by reference in its entirety.
  • the compound of the invention is not compound 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 32, 38, 47, 61, 71, 74, or 76 as depicted in the Exemplification section (i.e., wherein the exclusion of each compound listed is intended to be considered as a separate embodiment).
  • the compounds of the invention are selected based on at least one of the biological assays described herein. In yet another embodiment, the compounds of the invention are selected based on at least two of the biological assays described herein. In yet another embodiment, the compounds of the invention are selected based on at least three of the biological assays described herein. In yet another embodiment, the compounds of the invention are selected based on at least four of the biological assays described herein. It should be understood that, based on the distinctions among the biological assays described herein, a compound of the present invention is useful in the methods of the present invention if the compound produces a positive result within at least one of these biological assays.
  • such analysis may include a ratios, e.g., therapeutic ratios, of data obtained within a single assay, e.g., % protective effect (e.g., greater than a 50% protective effect, wherein protective effect may be defined in one embodiment as seizure onset delay multiplied by 100% and divided by 180), or between the data obtained from two or more different assays.
  • % protective effect e.g., greater than a 50% protective effect, wherein protective effect may be defined in one embodiment as seizure onset delay multiplied by 100% and divided by 180
  • the selection may be based on % protective effect, and the results of the PTZ induced seizure model, MES-seizure model, and the neurotoxicity screen.
  • the compound of the invention is not neurotoxic
  • the invention is directed to a method of treating, preventing, or inhibiting a seizure-associated state, e.g., epileptogenesis or ictogenesis, in a subject comprising administering to a subject in need thereof a sufficient amount of a compound of the present invention, wherein the compound is not neurotoxic, such that the seizure-associated state is treated, prevented, or inhibited.
  • the present invention is further directed, in particular embodiment to the use of prodrugs which are converted in vivo to the therapeutic compounds of the invention.
  • Such prodrugs can be used to alter the biodistribution (e.g., to allow compounds which would not typically cross the blood-brain barrier to cross the blood-brain barrier) or the pharmacokinetics of the therapeutic compound.
  • an anionic group e.g., a. carboxylate or sulfonate
  • carboxylate or sulfonate ester is administered to a subject, the ester is cleaved, enzymatically or non-enzymatically, to reveal the anionic group.
  • Such an ester can be cyclic, e.g. , a lactone or sultone, or two or more anionic moieties may be esterified through a linking group.
  • An anionic group can be esterified with moieties (e.g., acyloxymethyl esters) which are cleaved to reveal an intermediate compound which subsequently decomposes to yield the active compound.
  • an anionic moiety can be esterified to a group which is actively transported in vivo, or which is selectively taken up by target organs. The ester can be selected to allow specific targeting of the therapeutic moieties to particular organs.
  • the prodrug is a reduced form of an anionic group, e.g., a carboxylate or sulfonate, e.g., an alcohol or thiol, which is oxidized in vivo to the therapeutic compound.
  • an anionic group e.g., a carboxylate or sulfonate, e.g., an alcohol or thiol, which is oxidized in vivo to the therapeutic compound.
  • an additional embodiment of the invention is directed to the methods of preparation of compounds of the invention in accordance with the synthetic preparations described herein, e.g., in the Exemplification section.
  • the scope of the invention includes novel reactions described herein, which may include, without intending to be limiting, reactions of uracil and benzyl bromide, synthesis of 6- substituted uracils or thiouracils via cyclization, reaction of 6-methyluracil and p- nitrobenzyl bromide, coupling reactions of N-mono- and N-di-benzyl-5-iodouracil and hexyne, bromination of 6-methyl-3-para-nitrobenzyluracil, iodination of 6-methyl-l, 3- bis-para-nitrobenzyluracil, bromination of 6-methyluracil, benzoylation reactions, alkylation of benzoyl-protected uracil analogs, or synthesis of biuracil analogs via addition of sulf
  • the scope of the present invention also includes the extension of the reactions described herein to other systems, e.g., indoles and pyrroles, which contain the enamine moiety.
  • Compounds useful in the present invention may also include carrier or targeting moieties which allow the therapeutic compound to be selectively delivered to a target organ or organs.
  • the compound may include a moiety capable of targeting the compound to the brain, by either active or passive transport (a "targeting moiety").
  • the carrier molecule may include a redox moiety, as described in, for example, U.S. Patent Nos. 4,540,564 and 5,389,623.
  • an anti-ictogenic and/or anti- epileptogenic pharmaceutical composition of the present invention can include a sufficient amount of one or more of the compounds described herein effective to inhibit a seizure-associated state, such as epilepsy in a subject in need thereof, and a pharmaceutically acceptable carrier.
  • Ictogenesis may be treated, prevented, or inhibited in a subject in need thereof by administering to the subject a sufficient amount of one or more of the compounds described herein, such that ictogenesis is treated, prevented, or inhibited in the subject.
  • epileptogenesis may be treated, prevented, or inhibited in a subject in need thereof by administering to the subject a sufficient amount of one or more of the compounds described herein, such that epileptogenesis is treated, prevented, or inhibited in the subject.
  • the present invention provides pharmaceutically acceptable compositions which comprise a therapeutically-effective amount of one or more of the compounds described above, formulated together with one or more pharmaceutically acceptable carriers (additives) and/or diluents.
  • the pharmaceutical compositions of the present invention may be specially formulated for administration in solid or liquid form, including those adapted for the following: (1) oral administration, for example, drenches (aqueous or non-aqueous solutions or suspensions), tablets, boluses, powders, granules, pastes for application to the tongue; (2) parenteral administration, for example,, by subcutaneous, intramuscular or intravenous injection as, for example, a sterile solution or suspension; (3) topical application, for example, as a cream, ointment or spray applied to the skin; (4) transdermal application, for example, via a "patch,” for example, using a lotion, cream, solution, gel or solid ; or (5) intravaginally or intrarectally, for example, as a pessary,
  • the invention pertains to a pharmaceutical composition
  • a pharmaceutical composition comprising a sufficient amount of a compound of the present invention to treat, prevent, or inhibit a seizure-associated state, e.g., epilepsy, in a subject in need thereof, and a pharmaceutically acceptable carrier.
  • the pharmaceutical composition comprises a compound that is anti-ictogenic and/or anti-epileptogenic.
  • the invention further provides a kit which includes a container of a compound of the invention and instructions for administering a sufficient amount of the compound to a subject in need thereof such that a seizure-associated state (e.g., epileptogenesis) is treated, prevented, or inhibited in the subject.
  • a seizure-associated state e.g., epileptogenesis
  • the kits of the invention provide convenient means for administering the compounds of the invention.
  • the kit includes a sufficient amount of the compound, e.g., in unit dosage form.
  • the invention is directed to a kit for treating, preventing, or inhibiting a seizure-associated state in a subject comprising a compound of the present invention, and instructions for administering a sufficient amount of the compound to a subject, such that the seizure-associated state is treated, prevented, or inhibited.
  • the invention is directed to a packaged pharmaceutical composition for treating, preventing, or inhibiting a seizure-associated state in a subject comprising a container holding a compound of the present invention; and instructions for using the compound for treating, preventing, or inhibiting the seizure-associated state in a subject.
  • the invention is directed to a kit for treating, preventing, or inhibiting ictogenesis in a subject comprising a compound of the present invention, and instructions for administering a sufficient amount of the compound to a subject, such that ictogenesis is treated, prevented, or inhibited.
  • the invention pertains to a kit for treating, preventing, or inhibiting epileptogenesis in a subject comprising said compound of the present invention, and instructions for administering a sufficient amount of the compound to a subject, such that epileptogenesis is treated, prevented, or inhibited.
  • Another embodiment of the invention is a packaged pharmaceutical composition for treating, preventing, or inhibiting ictogenesis in a subject comprising a container holding a compound of the present invention; and instructions for using the compound for treating, preventing, or inhibiting ictogenesis in a subject.
  • the invention is directed to a packaged pharmaceutical composition for treating, preventing, or inhibiting epileptogenesis in a subject comprising a container holding a compound of the present invention; and instructions for using the compound for treating, preventing, or inhibiting epileptogenesis in a subject.
  • the compounds of the invention may be administered to subjects in a biologically compatible form, suitable for pharmaceutical administration in vivo.
  • biologically compatible form, suitable for administration in vivo describes a compound to be administered where any toxic effects are outweighed by the therapeutic effects of the compound.
  • Administration of a therapeutically active amount of the therapeutic compositions of the present invention is defined as an amount effective, at dosages and for periods of time necessary to achieve the desired result.
  • a therapeutically active amount of a compound of the invention may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of compound to elicit a desired response in the individual. Dosage regimes may be adjusted to provide the optimum therapeutic response. For example, several divided doses may be administered daily or the dose may be proportionally reduced as indicated by the exigencies of the therapeutic situation.
  • the active compound may be administered in a convenient manner such as by injection (subcutaneous, intravenous, etc.), oral administration, inhalation, transdermal application, or rectal administration.
  • the active compound may be coated in a material to protect the compound from the action of enzymes, acids and other natural conditions which may inactivate the compound.
  • a compound of the invention can be administered to a subject in an appropriate carrier or diluent, co-administered with enzyme inhibitors or in an appropriate carrier such as liposomes.
  • pharmaceutically acceptable carrier as used herein is intended to include diluents such as saline and aqueous buffer solutions.
  • Liposomes include water-in-oil-in-water emulsions as well as conventional liposomes (Strejan et ah, (1984) J Neuroimmunol 7:27).
  • the active compound may also be administered parenterally or intraperitoneally.
  • Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations may contain a preservative to prevent the growth of microorganisms.
  • compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion.
  • the composition must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi.
  • the pharmaceutically acceptable carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyetheylene glycol, and the like), and suitable mixtures thereof.
  • the proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.
  • Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like.
  • isotonic agents for example, sugars, polyalcohols such as manitol, sorbitol, sodium chloride in the composition.
  • Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate and gelatin.
  • Sterile injectable solutions can be prepared by incorporating active compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filter sterilization.
  • dispersions are prepared by incorporating the active compound into a sterile vehicle which contains a basic dispersion medium and the required other ingredients from those enumerated above.
  • the preferred methods of preparation are vacuum drying and freeze-drying which yields a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.
  • the composition may be orally administered, for example, with an inert diluent or an assimilable edible carrier.
  • pharmaceutically acceptable carrier includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, use thereof in the therapeutic compositions is within the scope of the present invention.
  • Supplementary active compounds can also be incorporated into the compositions. It is especially advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage.
  • Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the mammalian subjects to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier.
  • the specification for the dosage unit forms of the invention are dictated by and directly dependent on (a) the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of compounding such an active compound for the therapeutic treatment of individuals.
  • the invention provides libraries of compounds of the invention, and methods of preparing such libraries.
  • the synthesis of combinatorial libraries is well known in the art and has been reviewed ⁇ see, e.g., Gordon et al., J. Med. Chem. 37:1385-1401 (1994)).
  • the invention includes methods for synthesis of combinatorial libraries of compounds of the invention.
  • Such libraries can be synthesized according to a variety of methods. For example, a "split-pool" strategy can be implemented to produce a library of compounds.
  • the library of immobilized compounds can then be washed to remove impurities.
  • the immobilized compounds can be cleaved from the solid support to yield a compound of the present invention.
  • a "diversomer library” is created by the method of Hobbs, DeWitt et al. (Proc. Natl Acad. Sci. U.S.A. 90:6909 (1993)). After creation of the library of compounds, purification and workup yields a soluble library of substituted compounds of the present invention.
  • synthesis methods including the "tea-bag” technique of Houghten et al., Nature 354:84-86 (1991), can also be used to synthesize libraries of compounds according to the subject invention.
  • Combinatorial libraries can be screened to determine whether any members of the library have a desired activity, and, if so, to identify the active species. Methods of screening combinatorial libraries have been described (see, e.g., Gordon et al., J Med. Chem., op. cit.).
  • Soluble compound libraries can be screened by affinity chromatography with an appropriate receptor to isolate ligands for the receptor, followed by identification of the isolated ligands by conventional techniques (e.g., mass spectrometry, NMR, and the like).
  • Immobilized compounds can be screened by contacting the compounds with a soluble receptor; preferably, the soluble receptor is conjugated to a label (e.g., fluorophores, colorimetric enzymes, radioisotopes, luminescent compounds, and the like) that can be detected to indicate ligand binding.
  • a label e.g., fluorophores, colorimetric enzymes, radioisotopes, luminescent compounds, and the like
  • immobilized compounds can be selectively released and allowed to diffuse through a membrane to interact with a receptor.
  • Exemplary assays useful for screening the libraries of the invention are known in the art (see, e.g., Gordon et al., J. Med. Chem. 37:1385-1401 (1994)).
  • Combinatorial libraries of compounds can also be synthesized with "tags" to encode the identity of each member of the library.
  • this method features the use of inert, but readily detectable, tags, that are attached to the solid support or to the compounds.
  • tags When an active compound is detected such as by one of the techniques described above, the identity of the compound is determined by identification of the unique accompanying tag.
  • This tagging method permits the synthesis of large libraries of compounds which can be identified at very low levels.
  • the libraries of compounds of the invention contain at least 30 compounds, more preferably at least 100 compounds, and still more preferably at least 500 compounds. In particular embodiments, the libraries of compounds of the invention contain fewer than 10 9 compounds, more preferably fewer than 10 8 compounds, and still more preferably fewer than 10 7 compounds.
  • a library of compounds is preferably substantially pure, i.e., substantially free of compounds other than the intended products, e.g., members of the library.
  • the purity of a library produced according to the methods of the invention is at least about 50%, more preferably at least about 70%, still more preferably at least about 90%, and most preferably at least about 95%.
  • the libraries of the invention can be prepared as described herein.
  • at least one starting material used for synthesis of the libraries of the invention is provided as a variegated population.
  • the term "variegated population”, as used herein, refers to a population including at least two different chemical entities, e.g., of different chemical structure.
  • a "variegated population" of compounds of the present invention would comprise at least two different compounds of the present invention.
  • Use of a variegated population of linkers to immobilize compounds to the solid support can produce a variety of compounds upon cleavage of the linkers.
  • Libraries of the invention are useful for, inter alia, drug discovery.
  • a library of the invention can be screened to determine whether the library includes compounds having a pre-selected activity, e.g., anti-epileptogenic or anticonvulsant activity.
  • TLC Thin layer chromatography
  • 6-substituted uracils or thiouracils 5-bromo-l, 3- dibenzyluracil, 6-methyl-l, 3-bis-para-nitrobenzyluracil, 5-bromo-l-para- nitrobenzyluracil, 5-bromo-6-(2-hydroxy-2-phenylethyl)uracil, 5-(hydroxymethyl)uracil and 5-(bromomethyl)uracil, 5-(benzyloxymethyl)uracil, 5-(benzylthiomethyl)uracil, 5- (phenylaminomethyl)uracil, S-benzoyl-S-bromo-l-para-nitrobenzyluracil, 1 ,3-dibenzoyl- 5-bromo-6-methyluracil, 3-benzoyluracil, 5-iodo-6-methyl-l ,3-bis-para- nitrobenzyluracil, 1 ,
  • Particular compounds of the invention also include the following compounds, numbered 1 -80, each of which is considered a separate embodiment of the invention. Moreover, the listing of the compounds below is intended merely as a convenience, and not necessarily as imparting the characteristic of a group. Analogues of the following compounds are also within the scope of the present invention, including acceptable salts, e.g., pharmaceutically acceptable salts, and esters thereof, i.e., wherein the ester group may be added as a substituent to any position available for substitution and that would not affect the ability of the compound to perform its intended function.
  • a series of l-alkyl-5-bromouracils were prepared in a one pot-synthesis by direct reaction between alkyl bromides and uracil under basic conditions in DMSO solvent only at 85°C. Under the same reaction conditions, when other solvents such as ethanol, acetonitrile, THF or DMF are used, the regioselectivity is lost, and no bromination is observed. Moreover, in attempt to understand the role of DMSO and the mechanism that lead to such bromination and regioselectivity the following reactions (equations 2 - 5) were performed and analyzed.
  • R Benzyl, Isopropyl, Isobutyl, cyclohexylmethyl, p-mtrobenzyl
  • R 6 H 1 CH 3
  • the mechanism in the scheme shown below suggests that the bromodimethylsulfoxonium salt in equation (5) is the predominant brominating agent while the benzylating agent may primarily be the intermediate (3) in order to account for N-I alkylation preferences for C-6 unsubstituted uracils.
  • 6-methyluracil under these reaction conditions, undergoes bromination but no alkylated product is formed.
  • the alkyl halide or the molecular bromine cannot be completely excluded as possibilities in the alkylation or bromination reactions since, in some instances, N-disubstituted uracils are obtained as minor products.
  • reactions for the synthesis of l-alkyl-5-bromouracils were monitored by TLC, which revealed that bromination is completed before any alkylation occurs.
  • the intermediate (3) could possibly be a bromine source in the presence of nucleophilic uracil. In this case the reaction would proceed as shown in the scheme below. Then, the resulting 5-bromouracil would be benzylated after deprotonation with the base.
  • the compound (3a) was prepared from l,3-dibenzyl-5-iodouracil (1.8 g, 4.3 mmol), cuprous iodide (25.0 mg), [bis-(triphenylphosphine)]-palladium chloride (26.3 mg) and 1-hexyne (1.0 mL, 8.7 mmol) at 40 0 C for two hours. After discoloration of the organic phase (CHCl 3 ) with activated Norit charcoal, and purification by flash chromatography (CHCl 3 ), the compound (3a) was obtained as a fine powder (827.2 mg, 51.7%). Like (2a), the product (3a) was also non-fluorescent under UV light.
  • Iodomonochloride (0.7 g, 4.3 mmol) was added to an ice pre-cooled yellow solution of 6-methyl-l,3-6 ⁇ -/? ⁇ 7r ⁇ -nitrobenzyluracil (1.0 g, 2.5 mmol) in dichloromethane (15.0 mL) resulting in the formation of a dark red solution. This solution was stirred at 0 0 C for 2 hours, then at 45 0 C for 16 hours in an attempt to drive the reaction to completion.
  • Pre-cooled liquid bromine (4.1 mL, density 3.1) was slowly added to an ice cold suspension of 6-methyluracil in aqueous sodium hydroxide solution (1.0 M, 80.0 mL), and the resulting mixture was vigorously stirred in an ice bath for 1 hour, and then allowed to warm to room temperature.
  • the reaction mixture was heated to 5O 0 C, and bromine was slowly added until a persistent yellow color was obtained.
  • the pale yellow reaction mixture was refluxed until the yellow color disappeared, resulting in a white slurry.
  • the reaction mixture was cooled in ice bath; filtered; washed with water, ethanol and ether; and dried under vacuum overnight to afford the compound 5-bromo-6-methyluracil in quantitative yield.
  • This compound was prepared following the procedure for alkylation of benzoyl- protected uracils from the reaction of 3-benzoyluracil (4.1 g, 19.0 mmol), sodium hydride (504 mg, 21.0 mmol), and benzyl bromide (21.0 mmol) for a total time of 12 hours. After ethyl acetate-water extraction, the aqueous phase was discarded and the organic layer was dried over sodium sulfate. The residue from the organic phase was purified by column chromatography (ethyl acetate / hexanes: 2:3) affording the product 3-benzoyl-l-benzyluracil as white powder (4.8 g, 82.1%).
  • Compounds (2) and (3) were prepared in 75.8% and 85.6% yield respectively following the literature procedure by Knochel et al. [102].
  • Compound (2) was obtained from the reaction of uracil and formaldehyde (37% aqueous solution) in the presence of barium hydroxide.
  • Compound (3) was prepared by refluxing compound (2) with concentrated HBr.
  • the present invention also includes two efficient processes for the conversion of uracil derivatives (1) into their corresponding 5,5'-methylene- ⁇ w-uracils (2) using readily available aldehydes or sulfoxides.
  • reaction conditions (A) the product (2) is readily obtained from an alcohol intermediate upon dehydration while conditions (B) yield the same product from a sulfide intermediate following a loss of the mercaptan HSR7 via elimination or nucleophilic substitution.
  • the mechanism may be represented by the following:
  • the solvent is removed in vacuo and the resulting slurry is treated accordingly to isolate pure products (3) and (4).
  • sulfoxides of product (3) were also isolated as minor products.
  • the sulfide oxidation may occur on the column since the sulfoxide is not observed on TLC in the course of reaction.
  • valeraldehyde was also found to be slower and requiring more equivalents of aldehyde substrate to yield the expected uracil derivative 2m (Table 2A) in addition to the alkene product 9 (below) that may have resulted from the dehydration of the alcohol 6m (Table 2A).
  • 10 derived from the addition of DBU followed by the conversion of the imine to carbonyl by hydrolysis.
  • the following reaction may be performed. Under neutral conditions, a solution of uracil (4.1 g, 36.6 mmol) in DMSO (20.0 mL) was gently refluxed for 20 hours resulting into a white slurry. The reaction mixture was cooled to room temperature, and the solid filtered out, washed with acetone and dried overnight under vacuum to yield the compound 4a (Table 2) (2.5 g, 57.2%) as a white powder. The filtrate was concentrated by distillation under reduced pressure and the resulting slurry was added to boiling ethanol containing 5.O g of silica gel. The mixture was vigorously stirred for 15 minutes, and then prepared for chromatographic separation.
  • the compound 4f (Table 2) was prepared according to the acid-catalyzed procedure from benzyl sulfoxide (4.69 g, 20.4 mmol), 6-methyluracil (2.62 g, 21.6 mmol), and concentrated hydrochloric acid (50.0 mL) at 9O 0 C. After 12 hours of reaction, the solvent was removed on a rotary evaporator and the resulting slurry was refluxed in methanol (50 mL, 10 minutes), filtered to afford a white solid that was washed with chloroform, then ether, and dried under vacuum overnight to yield the product 4f (Table 2) as a white powder (1.81 g, 51.2%). Mp: 352 - 354 0 C (decomposes).
  • the compound (5a) (above) was prepared by gently refluxing the mixture of 5-methylthiomethyluracil (622.3 mg, 3.62 mmol), 1-benzyluracil (734.5 mg, 3.64 mmol) and concentrated hydrochloric acid (17.0 rnL). After 12 hours, the solvent was completely removed on a rotary evaporator, and the resulting residue was stirred in hot DMSO (9O 0 C), and then filtered to remove the insoluble solids found to be a mixture of (5a) and its debenzylated product. Acetone was added to the hot DMSO filtrate until cloudy, and then allowed to slowly cool down.
  • Compound 30a was prepared by gently refluxing the mixture of 5- methylthiomethyluracil (622.3 mg, 3.6 mmol), 1-benzyluracil (734.5 mg, 3.6 mmol) and hydrochloric acid (12 M, 17 mL). After twelve hours, the solvent was completely removed on a rotary evaporator, and the resulting residue was stirred in hot DMSO (10 mL, 90 0 C), and then filtered. Acetone was added to the hot DMSO filtrate until cloudy, and then allowed to slowly cool to room temperature. The resulting crystals were filtered, washed with acetone then ether, and dried overnight under vacuum to afford the compound 30a as white powder (218.2 mg, 18%).
  • Compound 2j was prepared according to the DBU-catalyzed reaction procedure starting from ortho-methoxybenzaldehyde (1.8 g, 13.6 mmol), uracil (1.3 g, 12.4 mmol), and DBU (1.8 mL, 12.0 mmol) in dry DMF (10 mL). After six hours of reaction, the solvent was removed on a rotary evaporator and the resulting slurry was treated as for the typical procedure. The obtained crude product was recrystallized from methanol and ether to afford the pure product 2j as a white powder (1.8 g, 86% yield). Mp: >300°C (decomposed).
  • Compound 21 was prepared according to the DBU-catalyzed reaction procedure starting from para-methoxybenzaldehyde (1.6 g, 11.5 mmol), uracil (1.3 g, 11.4 mmol), and DBU (1.7 mL, 11.0 mmol) in dry DMF (10 mL). After six hours of reaction, the solvent was removed on a rotary evaporator and the resulting slurry was treated as for the typical procedure. The obtained crude product was recrystallized from methanol to afford the pure product 21 as a white powder (1.5 g, 79% yield). Mp: >300°C (decomposed).
  • Compound 2m was prepared according to the DBU-catalyzed reaction procedure starting from valeraldehyde (1.8 g, 20.7 mmol), uracil (2.3 g, 20.5 mmol), and DBU (3.1 mL, 20.7 mmol) in dry DMF (20 mL). After twenty hours of reaction, uracil was still present in the reaction mixture; thus an extra 1 mL of valeraldehyde was added and reaction continued for two more hours to complete the reaction of uracil. The solvent was removed on a rotary evaporator and the resulting slurry was treated as for the typical procedure. The obtained crude product was recrystallized from methanol to afford the pure product 2m as a white powder (1.5 g, 49% yield).
  • Compound 2n was prepared according to the DBU-catalyzed reaction procedure starting from 2-thiophene-carboxaldehyde (1.3 g, 12.0 mmol), uracil (1.3 g, 11.4 mmol), and DBU (1.8 mL, 12.0 mmol) in dry DMF (10 mL). After twelve hours of reaction, the solvent was removed on a rotary evaporator and the resulting slurry was treated as for the typical procedure. The obtained dark red crude product was dissolved in aqueous sodium hydroxide (20 mL, 1 M) and the resulting solution was treated with activated charcoal under reflux for thirty minutes.
  • Compound 2o was prepared according to the DBU-catalyzed reaction procedure starting from 2-furan-carboxaldehyde (1.7 g, 18.1 mmol), uracil (2.0 g, 18.0 mmol), and DBU (2.6 mL, 17.4 mmol) in dry DMF (10 mL). After four hours of reaction, the solvent was removed on a rotary evaporator and the resulting slurry was treated as for the typical procedure. The obtained dark red crude product was dissolved in aqueous sodium hydroxide (20 mL, 1 M) and the resulting solution was treated with activated charcoal under reflux for thirty minutes.
  • Compound 2x was prepared according to the DBU-catalyzed reaction procedure starting from benzaldehyde (210.0 mg, 1.98 mmol), 3-benzyluracil (401.3 mg, 2.0 mmol), and DBU (300 ⁇ L, 2.0 mmol) in dry DMF (10 mL). After twelve hours of reaction, the solvent was removed on a rotary evaporator and the resulting dark slurry was treated as for the typical procedure. The obtained crude product was dissolved in acetone and the resulting solution was treated with the decolorizing activated charcoal (Norit A) under reflux for thirty minutes.
  • the decolorizing activated charcoal Neorit A
  • the obtained powder was mixed with hydrochloric acid (25 mL, 1 M) and stirred at 60°C for one hour. After cooling to room temperature, the fine precipitate was filtered, washed with water, acetone, and ether to afford the pure product 10 (2.6 g, 81% yield) as a white powder after overnight drying under vacuum. Mp >300 (decomposed).
  • Compound 11 was prepared according to the selective Nl-benzylation protocol for 5,5'-methylene-bis-uracil derivatives starting from the uracil derivative 10 (2.0 g, 5.4 mmol), BSA (5.3 mL, 21.7 mmol), and benzyl bromide (1.5 mL, 12.6 mmol) in dry acetonitrile (20 mL). After two hours, the reaction was complete and the work-up was performed according to the selective Nl-benzylation protocol. Recrystallization from methanol afforded the pure compound 11 as a white powder (2.7 g, 87% yield). Mp: 155-158 0 C.
  • Epilepsy is a very complex disorder of the brain and its symptoms cover a wide spectrum of seizure types and subtypes varying from simple partial seizures to generalized seizures.
  • testing of the compounds described herein is performed in a variety of seizure models.
  • These models include pilocarpine induced-seizures (PIS) model, the spontaneous recurrent seizure (SRS) model, the neurotrauma or brain trauma model (BTM), maximal electroshock induced-seizures (MES), subcutaneous pentylenetetrazol induced-seizures (PTZ) model and the hippocampal kindling seizure model.
  • PIS Pilocarpine induced-seizure
  • the test compound is dissolved in a mixture of saline (NaCl 0.9% by mass) and DMSO.
  • concentration of DMSO is normally 10% by volume. However, in cases where the test compound is very insoluble in this solvent mixture, the concentration of DMSO may be increased to a maximum of 50%.
  • test compound 100 mg/kg i.p. in saline/10% DMSO
  • SRS spontaneous recurrent seizures
  • Diazepam is administered (4 mg/kg i.p.) to stop the seizures
  • Test compound is injected I/day for 7 days (20 mg/kg i.p.) • Three weeks later, the animals are videotaped for 8 hours/day, 5 days/week for 2 months
  • BTM is a model that attempts to mimic the post-traumatic epilepsy, which is thought to result from the cascade of biochemical phenomena that follows a head (brain) injury.
  • the solution of the test compound is prepared as in PIS or SRS models, and the biological test is performed as follows:
  • Test compound is injected I/day for 7 days (20 mg/kg i.p.)
  • the numbers of seizures are counted from the videotape viewed by individuals blinded to the test drug/control information •
  • the animals are anesthesized (sodium pentobarbital), 5 mL of blood collected from the aorta
  • the MES model is described in, e.g., "Molecular and Cellular Targets for Anti- Epileptic Drugs” G. Avanzini, et al. (1997) John Libbey & Company Ltd., pp 191-198; Chapter 16, "The NIH Anticonvulsant Drug Development (ADD) Program: preclinical anticonvulsant screening project," by James P. Stables and Harvey J. Kupferberg.
  • seizures are induced in animals via electrical stimulation following the established NIH/ ADD program protocol.
  • Corneal electrodes primed with a drop of the electrolyte solution (9 % aqueous sodium chloride) are applied to the animal eyes and an electrical stimulus (50 mA in mice, 150 mA in rats; 60 Hz) is delivered for 0.2 seconds at the time of peak effect of the test compound.
  • the animal is restrained by hand and released at the moment of stimulation to allow observation of the entire seizure.
  • the test compound is considered to have offered protection against MES- induced seizure spread if there is abolition of the hind-leg tonic extensor component i.e. the hind-leg tonic extension does not exceed a 90° angle to the plane of the body.
  • the convulsant dose of pentylenetetrazole (85 mg/kg in mice and 70 mg/kg in rats) is injected at the time of peak effect of the test compound. Then, the animals are isolated and observed for a period of 30 minutes to see if seizures occur. Absence of clonic spasms persisting for five seconds or longer is an indication that the test compound may have the ability to increase the pentylenetetrazole induced-seizure threshold.
  • Hippocampal kindling seizure model This model is particularly useful as it, not only, provides an experimental model of seizures but also offers a means of studying the complex brain networks that may contribute to the spread and generalization of seizures from the focus, i.e., epileptogenesis.
  • Biological testing using hippocampal kindling seizure model follows the Lothman procedure (Lothman, et al, Epilepsy Res., 1988, 2:366) or the Silver procedure (Silver, et al., Ann. Neurol. (1991) 29:356). See also, Lothman, et al., Brain Res., 1994, 649:71, and Racine, et al., Clin. Neurophysiol., 1972, 32:281. This test is performed according to the procedure by Lothman [104] as summarized below:
  • a bipolar stimulating electrode is stereotactically implanted in the ventral hippocampus (AP-3.6, ML 4.9, DV-5.0 from dura, incisor bar +5) of adult male Sprague-Dawley rats (250 - 300 g) under ketamine-xylazine anesthesia.
  • Three anchor screws are attached to the skull and the electrode assembly anchored to the skull with dental acrylic cement. After the incision is closed with sutures, the animal receives a single dose of the Penicillin salt Bicillin, also known as Permapen, (60,000 units, i.m.) and is returned in its home cage to the animal quarters. Animals are kindled according to the Lothman & Williamson procedure [105].
  • test substance 50 mg/kg
  • BSS behavioral seizure score
  • ADD afterdischarge duration
  • Stage 2 Stage 1 plus head nodding
  • Stage 3 Stage 2 plus forelimb clonus • Stage 4: Stage 3 plus rearing
  • Stage 5 Stage 4 plus repeated rearing and falling
  • test-compounds are found to significantly lower the seizure score and decrease the afterdischarge duration, a dose-response study is initiated to quantify the drug- efficiency by varying the dose between 0 and 100% effect. Usually, the same group of kindled animals is used for multiple dose study as long as animals are permitted to dry- out for 4-5 days between studies. Test-compounds that lower the behavioral seizure score to 2 or less, and significantly decrease the afterdischarge duration are considered active or protective, and may be expected to be clinically useful against focal seizures that evolve to secondarily generalized seizures. Six Hertz psychomotor seizure test
  • MES and PTZ are very efficient in identifying compounds that may be useful for the treatment of the human generalized seizures, they may miss novel drugs that could be effective in the treatment of resistant partial epilepsy [107, 108].
  • An example of one such drug was Levetiracetam (UCB LO59), which was found to be active in the 6 Hz test but inactive in both MES and PTZ models. Seizures induced by the low frequency (6 Hertz) seem resistant to phenytoin and other sodium ion channel blockers, but sensitive to anticonvulsant drugs such as barbiturates, benzodiazepines, succinamides, and valproic acid that raise seizure threshold.
  • mice are pretreated intraperitoneally with 200 mg/kg of the test compound, and then challenged, after a certain interval of time (0.25, 0.5, 1.0, 4.0 h), with current (32 mA, 6 Hertz for 3 seconds) delivered through corneal electrodes to elicit a psychomotor seizure, which is characterized by a minimal clonic phase that is followed by sterotyped automatistic behaviors that resemble the aura observed for human patients with partial seizures. Subject animals that do not display these symptoms are considered protected, and the test compound active.
  • Neurotoxicity screen Preliminary neurotoxicity induced by the test compound may be characterized by neurologic abnormalities and poor performances in given tasks. For instance, the neurologic deficit may be indicated by the inability to maintain equilibrium for one minute in each of three trials on a knurled rod rotating at 6 rpm.
  • Other tests for neurotoxicity include the rotorod ataxia test, positional sense test, and gait and stance test. In the positional sense test, one hind leg is gently lowered over the edge of a table. The animal quickly reacts by returning the leg to the normal position. Failure to do so rapidly, indicates a neurologic deficit.
  • the neurologic deficit is indicated by a circular or zigzag gait, ataxia, and other abnormal events that may include abnormal spread of legs, body posture, tremor, hyperactivity, lack of exploratory behavior, somnolence, stupor, and catalepsy.
  • Biological testing results are indicated by a circular or zigzag gait, ataxia, and other abnormal events that may include abnormal spread of legs, body posture, tremor, hyperactivity, lack of exploratory behavior, somnolence, stupor, and catalepsy.
  • ED 50 (mg/kg, i.p.) (Low - High) TD 50 (mg/kg, i.p.)
  • Phenobarbital 12 (9 - 16) 11(7-18) 12 (8 - 17) 24(17-34)
  • Biological data in Tables 5 and 6 indicate that uracil analogs compare fairly well with established anticonvulsant drugs.
  • the analog (22) shows good biological activity in various animal models including MES, PTZ, Kindling, PIS, and virtually no neurotoxicity at dosage as high as 300 mg/kg, outperforming first-line drugs of choice such as phenytoin, clonazepam, carbamazepine, and phenobarbital. Showing activity in diverse biological tests, and offering high therapeutic indexes, the uracil-based drugs are expected to offer protection over a wide range of seizure types and sub-types, with very little risk for unwanted side effects.

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Abstract

Cette invention se rapporte à des procédés et à des agents utiles dans le traitement, la prévention ou l'inhibition d'états de crises, notamment dans le traitement, la prévention ou l'inhibition d'épileptogenèse et/ou d'ictogenèse. L'invention se rapporte également à des procédés de préparation d'agents anti-ictogéniques et/ou anti-épileptogéniques. Cette invention se rapporte également à des compositions pharmaceutiques destinées au traitement, à la prévention ou à l'inhibition d'états de crise, ainsi qu'à des kits renfermant les composés anti-ictogéniques et/ou anti-épileptogéniques décrits dans l'invention.
PCT/IB2005/004156 2004-10-12 2005-10-12 Composes de pyrimidine en tant qu'agents anti-ictogeniques et/ou agents anti-epileptogeniques Ceased WO2006070292A2 (fr)

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WO2009156182A3 (fr) * 2008-06-25 2010-03-11 Resprotect Gmbh Dérivés de l'uracile et leur utilisation
US8106064B2 (en) 2006-07-24 2012-01-31 Korea Research Institute Of Chemical Technology Pyrimidine-2,4-dione HIV reverse transcriptase inhibitors
US8119800B2 (en) 2007-12-21 2012-02-21 Korea Research Institute Of Chemical Technology Processes for preparing HIV reverse transcriptase inhibitors
US8334295B2 (en) 2007-06-29 2012-12-18 Korea Research Institute Of Chemical Technology Pyrimidine derivatives as HIV reverse transcriptase inhibitors
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WO2015066034A1 (fr) * 2013-10-28 2015-05-07 Icahn School Of Medicine At Mount Sinai Compositions et procédés pour moduler l'excitabilité neuronale et le comportement moteur
US9771332B2 (en) 2015-05-05 2017-09-26 Pfizer Inc. 2-thiopyrimidinones
RU2643520C1 (ru) * 2017-04-11 2018-02-02 Федеральное государственное бюджетное образовательное учреждение высшего образования "Волгоградский государственный медицинский университет" Министерства здравоохранения Российской Федерации ФГБОУ ВО ВолгГМУ МЗ РФ 1,3-Бис(4-метоксибензоил)пиримидин-2,4(1Н,3Н)-дион, обладающий разрывающей поперечные сшивки гликированных белков активностью
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US8106064B2 (en) 2006-07-24 2012-01-31 Korea Research Institute Of Chemical Technology Pyrimidine-2,4-dione HIV reverse transcriptase inhibitors
US8334295B2 (en) 2007-06-29 2012-12-18 Korea Research Institute Of Chemical Technology Pyrimidine derivatives as HIV reverse transcriptase inhibitors
US8354421B2 (en) 2007-06-29 2013-01-15 Korea Research Insitute Of Chemical Technology HIV reverse transcriptase inhibitors
US8119800B2 (en) 2007-12-21 2012-02-21 Korea Research Institute Of Chemical Technology Processes for preparing HIV reverse transcriptase inhibitors
WO2009156182A3 (fr) * 2008-06-25 2010-03-11 Resprotect Gmbh Dérivés de l'uracile et leur utilisation
US9181199B2 (en) 2008-06-25 2015-11-10 Resprotect Gmbh Uracil derivatives and use thereof
US9873673B2 (en) 2011-11-11 2018-01-23 Pfizer Inc. 2-thiopyrimidinones
US8835449B2 (en) 2011-11-11 2014-09-16 Pfizer Inc. 2-thiopyrimidinones
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US10870853B2 (en) 2013-10-28 2020-12-22 Icahn School Of Medicine At Mount Sinai Compositions and methods for modulating neuronal excitability and motor behavior
WO2015066034A1 (fr) * 2013-10-28 2015-05-07 Icahn School Of Medicine At Mount Sinai Compositions et procédés pour moduler l'excitabilité neuronale et le comportement moteur
US12163130B2 (en) 2013-10-28 2024-12-10 Icahn School Of Medicine At Mount Sinai Compositions and methods for modulating neuronal excitability and motor behavior
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US9771332B2 (en) 2015-05-05 2017-09-26 Pfizer Inc. 2-thiopyrimidinones
RU2643520C1 (ru) * 2017-04-11 2018-02-02 Федеральное государственное бюджетное образовательное учреждение высшего образования "Волгоградский государственный медицинский университет" Министерства здравоохранения Российской Федерации ФГБОУ ВО ВолгГМУ МЗ РФ 1,3-Бис(4-метоксибензоил)пиримидин-2,4(1Н,3Н)-дион, обладающий разрывающей поперечные сшивки гликированных белков активностью
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CN113045545A (zh) * 2021-03-23 2021-06-29 济南同路医药科技发展有限公司 一种咪唑斯汀有关物质a的制备方法

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