GB2440242A

GB2440242A - 1,2,4-Triazine derivatives as sodium channel blockers, antifolates and antimalarials

Info

Publication number: GB2440242A
Application number: GB0713564A
Authority: GB
Inventors: Michael Leach; Laurence Harbige; Dieter Riddall; Paul Barraclough
Original assignee: University of Greenwich
Current assignee: University of Greenwich
Priority date: 2006-07-13
Filing date: 2007-07-13
Publication date: 2008-01-23
Anticipated expiration: 2027-07-13
Also published as: CN104739839A; WO2008007149A3; EP2043656A2; JP5473598B2; CA2657577A1; AU2007274038A1; IL196469A0; CA2657577C; HK1129206A1; AU2007274038B2; CN101516377A; US20130005732A1; GB0613836D0; EP2043656B1; JP2009542792A; WO2008007149A2; US20090291954A1; GB0713564D0; IL196469A; ES2462927T3

Abstract

Use of compounds of formula (I) or a salt or solvate thereof as sodium channel blockers, antifolates, antimalarials or for use against mammalian cancers: wherein R1 is H or optionally substituted- alkyl, alkenyl, alkylaryl, alkylheterocyclyl; cycloalkyl; R2 to R6 are independently H, halogen, optionally substituted- alkyl, alkenyl, alkynyl or alkoxy. Preferred compounds include where R1 is an optionally substituted alkyl, aralkyl or heterocyclyl-alkyl group. In another aspect, compounds of formula (I) per se wherein R1 is optionally substituted- alkylaryl, alkylheterocyclyl; R2 is halogen, optionally substituted- alkyl, alkenyl, alkynyl or alkoxy; R3 to R6 are independently H, halogen, optionally substituted- alkyl, alkenyl, alkynyl or alkoxy. Some novel compounds where R1 is an aralkyl or heterocyclyl-alkyl are disclosed.

Description

NEW MEDiCAL USE OF TRIAZINE DERIVATIVES The present invention relates

to the use of triazine compounds as sodium channel blockers and antifolates and for preparation of medicaments for treatment of associated disorders.

US Patent No. 4,649,139 discloses compounds of the formula (A): R3 kIN R R5 y' y N 6 J

H N N 2 (A)

in which R' is C1..10 alkyl, C7lOalkenyl, C2.0 alkynyl or C31o cycloalkyl, any of which is optionally substituted, and R2 to R6 are independently selected from hydrogen, halogen, C1 alkyl, alkenyl, alkynyl or alkoxy (all optionally substituted by one or more of halogen, hydroxy and aryl), amino, mono-or di-substituted amino, alkenyloxy, acyl, acyloxy, cyano, nitro, aryl and alkylthio groups or any adjacent two of R2 to R6 are linked to form a (-CH=CH-CH=CH-) group. It is disclosed that these compounds are active in the treatment of cardiac disorders, and are particularly useful in the treatment of arrhythniias.

The present invention is based on the finding that compounds within formula (A) and certain novel derivatives thereof are potent sodium channel blockers and so are indicated to be useful as voltage dependent sodium channel blockers in treating disorders in mammals and particularly of value in the treatment of epilepsy, multiple sclerosis, glaucoma and uevitis, cerebral traumas and cerebral ischaernias, stroke, head injury, spinal cord injury, surgical trauma, neurodegenerative disorders, motorneurone disease, Alzheimers disease, Parkinsons disease, chronic inflammatory pain, neuropathic pain, migraine, bipolar disorder, mood, anxiety and cognitive disorders, schizophrenia and trigeminal autonomic cephalalgi as. Some conipounds also show anti!blate activity and so are indicated as of value as antifolates for the treatment of mammalian cancers and as antimalarials against plasmodium vivax and plasmodium falciparum malaria.

Accordingly, the present invention provides use of a compound of formula (1): R3 kIN RI R y y N 6k) H2N N NH (1) or a salt or solvate thereof in which R' is hydrogen, C11() alkyl, C21() alkenyl, C1 alkyl-aryl, C1 alkyl-heterocyclyl, or C 10 cycloalkyl, any of which is optionally substituted by halogen, halo C1 alkyl, C1 alkyl or C1 alkoxy; R2 to are independently selected from hydrogen, halogen, C1 alkyl, alkenyl, alkynyl or alkoxy (all optionally substituted by one or more of halogen, hydroxy and aryl), amino, mono-or di-substitutcd amino, alkenyloxy, acyl, acyloxy, cyano, nitro, aiyl and alkylthio groups; (a) as voltage dependent sodium channel blockers for the treatment of disorders in mammals, and particularly epilepsy, multiple sclerosis, glaucoma and uevitis, cerebral traumas and cerebral ischaemias, stroke, head injury, spinal cord injury, surgical trauma, neurodegenerative disorders, motorneurone disease, Alzheirners disease, Parkinsons disease, chronic inflammatory pain, neuropathic pain, migraine, bipolar disorder, mood, anxiety and cognitive disorders, schizophrenia and trigeminal autonornic cephalalgias, especially in humans; (b) as antifolates for the treatment of disorders in mammals, and particularly lbr treatment of mammalian cancers and as antimalarials against plasmodium vivax and plasmodium falciparum malaria, especially in humans.

The invention further includes the use of compounds of formula (1) for the preparation of a medicament for use as a sodium channel blocker or as an antifolate or as an antimalarial, especially for treatment of the individual disorders mentioned above.

As a C1io alkyl group, R1 is suitably an unsubstituted C1 alkyl group, typically methyl, ethyl, i-propyl, n-propyl, i-butyl or n-butyl.

As a C210 alkenyl group, R' may be an unsubstituted C2 alkenyl group, such as allyl.

As a C0 cycloalkyl group, R' is typically cyclohexyl, optionally substituted by one or niore halogen, haloalkyl or alkoxy groups, for example chioro, fluoro, tn fluoromethyl, methoxy or ethoxy.

As a C alkylaryl group, R' is typically benzyl in which the phenyl group is optionally substituted by one or more halogen, haloalkyl or alkoxy groups, for example chloro, fluoro, trilluorornethyl, trilluoromethoxy, niethoxy or ethoxy.

As a C13 alkyl-heterocyclyl, R' is suitably piperidine-niethyl, optionally N-substituted, or thienyl-niethyl, or furyl-methyl.

The R2 to R6-subslituted phenyl ring suitably contains one, two or three substituents..

R2 to R6 when other than hydrogen are preferably selected from halogen, halo alkyl or C1 alkoxy groups. Particularly preferred substitutions are 2,3 or 2,4 or 2,5 or or 3,5 or 2,3,5 di-or tn-halo (especially chloro and/or Iluro).

In a preferred class of compounds, R' is not hydrogen.

In another preferred class of compounds, R2 is not hydrogen.

In a further preferred class of compounds, both R' and R2 are not hydrogen.

In a preferred class of compounds of!brmula (I): R' is C11() alkyl, C210 alkenyl, C13 alkyl-aryl or C13 alkyl-hetcrocyclyl, any of which is optionally substituted by halogen, halo C1(,alkyl, C1alkyl or C1.,alkoxy; and R2 to R6 are independently selected from hydrogen and halogen.

In a favoured group of compounds having neuroprotective properties, R' is C alkyl, optionally substituted by CF3 for example methyl, ethyl, n-propyl, iso-butyl, n-butyl and trifluoropropyl, and R2 and R3, or R2 and R4, or R2 and R5, or R3 and R5, or R2, R3 and R5 are halo, especially chloro and/or fluoro.

Within formula (I) there is a group of compounds in which R' is hydrogen and R2 to R6 are independently selected from hydrogen, halogen, haloalkyl and haloalkoxy.

Within fommia (I) there is a group of compounds in which R' is alkyl, hydroxyalkyl, haloalkyl, heterocyclylalkyl, alkenyl, carboxamido, benzyl, benzyl substituted by halogen, alkyl, alkoxy, hydroxyalkyl, haloalky! or carboxamido and R2 to R6 are independently selected from hydrogen and halogen.

Within formula (I) there is a group of compounds in which R2 to are hydrogen and R' is hydrogen or alkyl Compounds of formula (1) which are novel form a further aspect of this invention.

In particular, compounds of formula (I) in which R' is optionally substituted C13 alkyl-heterocycl yl or optionally substituted C1 alkyl-aryl (excluding unsubstituted benzyl) are believed to be novel compounds.

Illustrative compounds of formula (I) are 5-amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2-isopropyl-I,2, 4-triazine; 5-arnino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2-n-propyl-1,2, 4-triazine; 5-amino-6-(2-pentyloxphenyl)-2,3-dihydro-3-imino-2-methyl-1,2,4-triazine; 5-amino-6-(2,3,5-trichlorophenyl)-2,3-dihydro-3-imino-2-methyl-I,2, 4-triazine; 5-amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2-methyl-1,2, 4-triazine; and 5-amino-6-(2,3-dichlorophenyl)-2,3dihydro-3-imino-2-benzyl-1,2,4-triazine; Further compounds of formula (I) include: 5-amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2-ethyl-I,2,4-triazine; 5-amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2-isopropyl-J,2, 4-triazine; 5-amino-6-(2,3-clichloropheny!)-2,3-dihydro-3-imino-2-n-propyl-J,2, 4-triazine; 5-anhino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2-isobutyl-1,2, 4-triazine; 5-amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2-n-butyl-1,2, 4-triazine; 5-aniino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-iniino-2-ally!-1,2, 4-triazine; 5-amino-6-(2,3,5-trichlorophenyl)-2,3-dihydro-3-imino-2-methyl-J,2, 4-triazine; 5-arnino-6-(2,3,5-trichlorophenyl)-2,3-dihydro-3-imino-2-propyl-1,2, 4-triazine; 5-amino-6-(2-lluoro,3-chlorophenyl)-2,3-dihydro-3-imino-2-methyl-1,2, 4-triazine; 5-arnino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2-(3,3, 3-trifluoropropyl)-1,2,4-triazine; 5(3)-amino-6-(2,4-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2-methyl-1, 2,4-triazine; 5(3)-aniino-6-phenyl-2,3(2,5)-dihydro-3(5)-irnino-2-methyl-1,2,4-triazine; 5(3)-amino-6-phenyl-2,3(2,5)-dihydro-3(5)-imino-2-ethyl-1,2,4-triazine; 5(3)-arnino-6-(2,5-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2-mcthyl-1, 2,4-triazine; 5(3)-arnino-6-(2,5-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2-ethyl-I, 2,4-triazine; 5(3)-arnino-6-(2,3,5-trichlorophenyl)-2,3(2,5)-dihydro-3(5) -imino-2-ethyl-1,2,4-triazine; 5(3)-amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2(2-fluoroethyl)-I,2,4-triazine; 5(3)-amino-6-(3,5-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-iniino-2-methyl-1, 2,4-triazine; 5(3)-arnino-6-(3,5-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2-ethyl-1, 2,4-triazine; 5(3)-arnino-6-(2,3,5-trichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2(2-fluoroethyl)- 1,2,4-triazine; 5(3)-aniino-6-(2,3,5-trichlorophenyl)-2,3(2,5)-dihydro-3(5)-irnino-2-(3,3, 3-tnfluoropropy!)-I,2,4-triazine; 5(3)-amino-6-(2,3,dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2-(2,2-di fluoroethy!)-I,2,4 triazine.

Another group of compounds of formula (1) includes: 3,5-diami no-6-(2, 5-dichiorophenyl)-1,2,4-triazine, 3,5-diamino-6-(3,5-dichlorophenyl)-1,2,4-lriazine, 3,5-diamino-6-phenyl-1,2,4-triazine, 3,5-diamino-6-(2,4-dichlorophenyl)-I,2,4-triazine, 3,5-diamino-6-(2-tri!luorornethoxyphenyl)-I,2,4-triazine.

Novel compounds of formula (I) include: 5-arnino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2-(2 -fluorophenyl-methy!)-I,2,4-triazine; 5-ami no-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino2(3 -fluorophenyl-methyl)-1,2,4-triazine; S-arnino-6-(2,3-dich!orophenyl)-2,3-cljhydro-3-imino-2(4 -Iluorophenyl-methyl)-I,2,4-triazine; 5-amino-6-(2,3-dichloroplienyl)-2,3-dihydro-3-imino-2-(2 3 -diii uoropheny!-methyl)-I,2,4-triazine 5-arnino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2-(3 -chlorophenyl-methy!)- 1,2,4-triazine 5-arnino-6-(2,3-dichloropheny!)-2,3.dihydro-3-jmino2(4 -chiorophenyl-methyl)-I,2,4-triazine; 5-arnino-6-(2,3-dichlorophenyl)-2,3-dihydro-3 -imino-2-(4 -methyiphenyl-methyl)-l,2,4-triazine; 5-arnino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2-(2 -methoxyphenyl-methyl)- 1,2,4-triazine; 5-arnino-6-(2,3-dichlorophenyl)-2,3-dihydro-3..inhjno-2(3 -methoxyphenyl-methy!)- 1,2,4-triazine; 5 -amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-irnino-2-(4 -methoxyphenyl-methyl)-I,2,4-triazine; 5-amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2-(2 -chiorophenyl-methyl)-I,2,4-triazine; 5-amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-irnino-2-(2 -tn fluorornethyl pheny!-methyl)-I,2,4-triazine; 5-arnino-6-(2,3 -dichiorophenyl)-2,3 -dihydro-3 -imino-2-(3 -tn fluoromethyiphenyl-methyl)-I,2,4-triazine; 5-arnino-6-(2,3-di chiorophenyl)-2,3-di hydro-3-imino-2-(4 -tn fluorornethylphenyl-methyl)-1,2,4-triazine; 5-amino-6-(2,3-dichloropheny!)-2,3-djhydno-3-jmino2(2 -fluoro-3 -tn Ii uorornethylphenyl-methyl)-I,2,4-triazine; 5-amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2-(3thienylniethyl) 1,2,4-triazine; 5-arnino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2-(3-furyl-rnethyl)-1, 2,4-triazine and S-arnino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2{N4,ocpiperidjn4yl methyl)-I,2,4-triazine; S-amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-inijno2(piperjdin4ylniethyl) 1,2,4-triazine.

The use of salts of the compounds of formula (I) form an aspect of this invention.

Preferred salts are pharmaceutically acceptable acid addition. Suitable pharmaceutically acceptable acid addition salts include those formed with both organic and inorganic acids, for example from hydrochloric, sulphuric, citric, tartaric, phosphoric, lactic, pyruvic, acetic, malonic, succinic, oxalic, fumaric, maleic, ox aloacetic, methanesuiphonic, p-toluenesulphonic, benzene-sulphonic, gI utanhic, naphthoic, and isethionic acids. Esylate (ethanesulfonate), edisylate (1,2-ethanesulfonate), malate, mandalate, benzoate, and salicylate salts are also suitable.

In preparation of the compounds of formula (I), the compound or its salt may be obtained as a solvate of the reaction or crystallisation solvent or a component thereof.

Use of such solvates forms another aspect of this invention. Suitable pharmaceutically acceptable solvates include hydrates.

The present invention includes within its scope use of all tautoniers, enantiomers and polymorphs of the compounds of formula (1), and of salts and solvates thereof The compounds of formula (I) may suitably be prepared by the procedures disclosed in the above-mentioned US Patent No. 4,649,139, the entire disclosure of which is incorporated herein by reference.

Conveniently, a compound of Formula II :II (II) in which R2 -R6 are as deiincd for formula (1) is reacted with a compound R1-Q (Ill) where RI is as defined for formula (I) and Q is a leaving group.

Suitable leaving groups include halogens and sulphonic acid derivatives, such as mesyl, tosyl, etc. The reaction suitably take places under conventional conditions in a solvent in which the compound of formula II is soluble at a convenient temperature (for example between 0 and 100 C. and niost conveniently at rooni temperature).

Compounds of formula II may be prepared by methods disclosed in EP 0 021121 A, the entire disclosure of which is incorporated herein by reference.

Salts of compounds of formula (1) may be obtained by the presence of a residual Q acid. Alternatively salts may be prepared by mixing the compound of formula (I) as the free base with a pharmaceutically acceptable acid in a suitable solvent, and removing the solvent to recover the salt, or crystallising the salt from the solvent.

In a further aspect, the present invention provides phamiaceutical compositions for the treatment of disorders such as epilepsy, multiple sclerosis, glaucoma and uevitis, cerebral traumas and cerebral ischaemias, stroke, head injury, spinal cord injury, surgical trauma, neurodegencrative disorders, motorneurone disease, Alzheiniers disease, Parkinsons disease, chronic inflammatory pain, neuropathic pain, migraine, bipolar disorder, mood, anxiety and cognitive disorders, schizophrenia and trigeminal autonomic cephalalgias; for treatment of mammalian cancers; and for treatment of malaria; comprising a conipound of formula (I), or a pharmaceutically acceptable salt or solvate thereo1 in admixture with a pharmaceutically acceptable carrier.

The compounds of formula (1) will be present in the compositions of the present invention in an effective unit dosage form, that is to say in an amount sufficient to be effective against the disorders in vivo.

The pharmaceutically acceptable carriers present in the compositions of the present invention may be materials conventionally used for the purpose of administering the medicament. These may be liquid or solid materials, which are otherwise inert or medically acceptable and are compatible with the active ingredients.

These pharmaceutical compositions may be given orally or parenterally, for example as a suppository, ointment, creanî, powder or trans-dermal patch. However, oral administration and intravenous injection of the compositions are preferred.

For oral administration, fine powders or granules will contain diluting, dispersing andlor surface active agents, and niay be presented in draught, in water or in a syrup, in capsules or sachets in the dry slate or in non-aqueous suspension wherein suspending agents may be included, or in a suspension in water or syrup. Where desirable or necessary, flavouring, preserving, suspending, or thickening agents can be included. Dry powders or granules may be compressed to form a tablet or contained in a capsule.

For injection, the conipounds may be presented in sterile aqueous injection solutions which may contain anti-oxidants or buffers.

The free base or a salt or solvate thereof may also be administered in its pure form unassociated with other additives in which case a capsule or sachet is the preferred carrier.

Alternatively the active compound may be presented in a pure form as an effective unit dosage, for instance compressed as a tablet or the like.

Other compounds which may be included are, for example, medically inert ingredients, e.g., solid and liquid diluents such as lactose, starch, or calcium phosphate for tablet or capsules; olive oil or ethyl oleate for soft capsules; and water or vegetable oil for suspensions or emulsions; lubricating agents such as tale or magnesium stearale; gelling agents such as colloidal clays; thickening agents such as gum tragacanth or sodium alginate; and other therapeutically acceptable accessory ingredients such as humectants, preservatives, buffers, and antioxidants which are useful as carriers in such formulations.

Tablets or other forms of presentation provided in discrete units may conveniently contain an amount of compound of formula I which is effective at such dosage or as a multiple of the same, for instance units containing 5 nig to 500 mg, usually around 10 rngto250mg.

The pharmaceutical compositions of the present invention may be prepared by the adniixture of a compound of formula (1) with a pharmaceutically acceptable carrier.

Conventional pharmaceutical excipients niay be admixed as required. Example of suitable formulations are give in the above- mentioned US Patent. No. 4,649,139.

The present invention provides a method of treatment of disorders in mammals that are susceptible to sodium channel blockers and antifolates, and particularly disorders such epilepsy, multiple sclerosis, glaucoma and uevitis, cerebral traumas and cerebral ischaemias, stroke, head injury, spinal cord injury, surgical trauma, neurodegenerative disorders, motorneurone disease, Alzheimers disease, Parkinsons disease, chronic inflammatory pain, neuropathic pain, migraine, bipolar disorder, mood, anxiety and cognitive disorders, schizophrenia and trigeminal autonomic cephalalgias; for Ireatnient of mammalian cancers; and for treatment of malaria; by the administration of a non-toxic effective amount of a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or a composition as hereinbefore defined.

As indicated above, the compounds of formula (I) are generally useful in treating such disorders by oral administration or intravenous injection.

The compounds of formula (1) are normally administered at a dose of' from 0.01 mg/kg to 20 mg/kg per day, preferably 0. 1 to 5.0 mg/kg per day. The dose range for adult humans is thus generally from 0.7 nig to 1400 mg/day and preferably 7 to 350 mg/day.

In view of' the known use in humans of structurally similar compounds such as lamotrigine, no major toxicity problems are anticipated in use of compounds of formula (I). However appropriate testing procedures should be carried out before clinical use.

The following Examples show the preparation of illustrative compounds of formula (1) and other compounds used in testing, as reported below.

Example 1 -Lamotrigine Larnotriginc -5(3)-amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-iniino-I,2, 4-triazine -is approved for human use as an anticonvulsant for treatment of epilepsy and is commercially available under the name LAM ICTAL (GSK). The preparation of lamotrigine is disclosed in European Patent No. 0021121.

Example 2

5-ami no-6-(2,3-dichlorophenyl)-2,3-di hydro-3-imino-2-methyl-1,2,4-triazine The preparation of the title compound free base is described in US Patent No. 4,649,139 (Example 1), by reaction of 3,5-diamino-6-(2,3-dichlorophenyl)-I,2,4-triazine with iodomethane. The methanesulfonate salt was prepared from the free base as follows.

Methyl methanesulfonate (0.50 g, 4.5 mmol), 3,5-diamino-6-(2,3-dichlorophenyl) - 1,2,4-triazine (0.50 g, 2.0 mmol) and dirnethylformamide (4 ml) were stirred and heated at 100 C for 10 nun. The solution was cooled, toluene (20 ml) added, and the mixture stirred for 0.5h. The solid was collected by filtration and recrystallised from propan-2-ol to give the methanesulfonate salt of the title compound as a white solid (0.40 g), mp 274-276 C.

Example 3

5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5) -imino-2-ethyl-1,2,4-triazine lodoethane (3.12 g, 0.02 mol) was added to a stirred suspension of 3,5-diamino-6-(2,3-dichlorophenyl)-1,2,4-triazine (2.56 g, 0.01 mol) in acetone (200 ml). The mixture was stirred at room temperature for 5 days, more ethyl iodide added (1.56 g, 0.01 mol) and stirring continued for a further 3 days. The solid was collected by filtration and then stirred in 40 ml of 18% ammonia solution. The solid (ca. 2.5 g) was removed by filtration, dried in vacuo and recrystallised from methanol to give 1.4 g (22 %) of the title compound as a white crystalline solid, mp 216-217 C 6-(500 MHz, dmso-d6) 1.21 (3H, 1, J = 7. 0 Hz, C-CH3), 3.90 (2H, q, I = 7.0 Hz, NCH2), 4.15 (IH, brpeak, NH), 6.2-7. 2 (2H, vbrpeak, NH2), 7.41 (2H, iii, aromatic H), 7.71 (IH, dd,1 8,2 Hz, aromatic H).

Methanesulfonate salt mp 255-260 C.

Example 4

5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5) -imino-2-isopropyl-1,2,4-triazine 2-lodopropane (2 nil, 3.4g, 0.02 mol) was added to a stirred suspension of 3,5-diamino-6-(2,3-dichlorophenyl)-I,2,4-triazine (2.56g. 0.01 mol) in acetone (200 nil). The mixture was stirred at rellux for 5 days, more 2-iodopropane (1 ml, 0.01 niol) added and refluxing continued for 2 days. After cooling, the solid was collected by filtration and then stirred with 0.88 aqueous ammonia (80 ml) for 0.5 h. The solid (ca. 2.5 g) was removed by filtration, dried in vacuo and recrystallised from methanol to give 1. 0 g (34 %) of the title compound as a pale yellow crystalline solid, mp 209-2 12 C.

8n (500 MHz, dmso-d6) 1.21 (6H, t, J= 7 Hz, CH3-C-CH3), 3.21 (3H, s, CH3OH), 4.15 (1H, brpeak, NH), 4.84 (1H, brpeak, CHN), 7.38-7.46 (2H, m, aromatic H), 7.71 (1H, dd, J = 8,2 HL, aromatic H). This compound is a methanol solvate.

Methanesulfonate salt mp 247-250 C.

Example 5

5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2(2-hydroxyethyl)-I,2,4-triazine 2-lodoethanol (3.44 g, 0.02 mol) was added to a stirred suspension of 3,5-diarnino-6-(2,3-dichlorophenyl)-1,2,4-triazine (2.56 g, 0.01 mol) in acetone (200 ml). The mixture was stirred at reflux for 6 days, cooled and the solid collected by filtration. The solid was stirred with 0.88 aqueous ammonia (100 ml) and the mixture stirred for 0.5 h. The solid (ca. 2.7 g) was removed by filtration, dried in vacuo and recrystallised from methanol to give 1.14 g (38 %) of the title conipound as a white crystalline solid, mp 217-218 C ö (500 MHz, dmso-&) 3.34 (3H, s, CH3OH), 3.68 (2H, brt, .1 6 Hz, OCH2), 3.96 (2H, iii, NCH), 5.5-7.0 (2H, vbrpeak, NH2), 7.36-7A6 (2H, m, aroniatic H), 7.71 (IH, dd, J = 8,2 Hz, aromatic H). This conipound is a methanol solvate Methanesulfonatc salt mp 242-245 C]

Example 6

5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5) -imino-2-propyl-1,2,4-triazine 1-lodopropane (3.4 g, 0.02 mol) was added to a stirred suspension of 3,5-diamino-6-(2,3-dichlorophenyl)-1,2,4-triazinc (2.56 g, 0.01 mol) in acetone (200 ml). The mixture was stirred at reflux for 2 days, more 1-iodopropane (1.7 g, 0.01 mol) added and refluxing continued for a further 24h. After cooling, the solid was collected by uiltration and then stirred with 0.88 aqueous ammonia (80 ml) for 0.5 h. The solid (ca. 3. 1 g) was removed by filtration, dried in vacuo and recrystallised from methanol-water (ca. 160 ml) to give 1.65 g (56 %) of the title compound as a white crystalline solid, mp 197-199 C.

n (500 MHz, dmso-d6) 0.88 and 0.91 (3H, 2 x t,J= 7Hz, C-CH3),l.64-1.74 (2H, m, C-CH2-C), 3.82 and 3.90 (2H, 2 x t, I = 7 Hz, NCH2), 6.2-7.4 (lH, vbrpeak, NH), 7.35-7.46 (2H, rn, aromatic H), 7.71 (IH, rn, aromatic H). Two tautomers are present inratio4:1.

Methanesulfonate salt mp 237-240 C

Example 7

5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5) -imino-2-isobutyl-1,2,4-triazine methanesulfonate 2-lodobutane (1.8 ml, 2.88 g, 0.016 mol) was added to a stirred suspension of 3,5-diamino-6-(2,3-dichlorophenyl)-1,2,4-triazine (1.28 g, 0.005 mol) in acetone (50 ml).

The mixture was stirred and heated at reflux for 4 days. More l-iodobutane (0.6 ml, 0.005 mol) was added and refluxing continued for I day. After cooling, the solid was collected by filtration and then stirred with 0.88 aqueous ammonia (80 ml) for 0.5 h. The resulting solid (ca.0.9 g) was removed by filtration, dried in vacuo. A portion (0.31 g) was stirred with methanesulfonic acid (0.10 g) in methanol (3.5 ml) and the mixture diluted with ether to give the mesylate of the title compound (0.22 g) as a white crystalline solid, of no sharp mp (decomp.> 230 C (500 MHz, dniso-d6) 0.92 (6H, d, J = 5.9 Hz, 2 x C-CH3), 2. 12 (1 H, m, CHMe2) 2.30 (3H, s, SCH3), 3.92 (2H, brs, NCH2), 7.54 (2H, m, aromatic H), 7.86 (IH, dd,J= 7.2, 2.5 Hz, aromatic H), 8.18 (IH, brs, NH, exchang.), 8.2-8.8 (2H, vbrpeak, NH2, exchange), 9.14 (IH, brs, NH, exchange.).

Example 8

5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5) -imino-2-butyl-1,2,4-triazine 1-lodohutane (2.3 ml, 3.68 g, 0.02 fbI) was added to a stirred suspension of 3,5-diarnino-6-(2,3-dichlorophenyl)-I,2,4-triazine (2.56 g, 0.01 mol) in acetone (200 nil). The mixture was stirred and heated at refiux for 4 days. More I-iodobutane (0.6 ml, 0.005 mol) was added and relluxing continued for 1 day. After cooling, the solid was collected by filtration and then stirred with 0.88 aqueous ammonia (80 ml) for 0.5 h. The resulting solid (ca.2.2 g) was renioved by filtration, dried in vacuo and recrystallised from methanol to give I. I g (35 %) of the title compound as a white crystalline solid, nip 175 C.

6-(500 MHz, dniso-d6) 0.89 (3H, t, I = 7 Hz, CH3), 1.31 (2H, hextet, I = 7 Hz, CH2Me), 1.64 (2H, pent, I = 7 Hz, CH2-C-Me), 3.86 (2H, 1, 1 = 7 Hz, NCH2), 6.2-7.2 (2H, vbrpeak, NH) 7.38 (IH, dd, I = 8,2 Hz, aromatic H), 7.43 (IH, t, I = 8 Hz, aromatic H), 7.70 (1 H, dd, I = 8,2 Hz, aromatic H).

Example 9

5(3)-Am ino-6-(2,3-dichlorophenyl)-2,3(2,5)-clihydro-3(5)-imino-2-phenylmethyl-1, 2,4-triazine hemi-methanesulfonate Benzyl chloride (0.92 ml, 1.01 g, 0. 008 mol) was added to a stirred suspension of 3,5-diamino-6-(2,3-dichlorophenyl)-1,2,4-triazjne (1.0 g, 0.004 mol) in acetone (50 ml). The mixture was stirred and heated at reflux for 3 days. More benzyl chloride (0.6 ml, 0.005 mol) was added and refluxing continued for 2 days. After cooling, the solid was collected by filtration and then stirred with 0.88 aqueous ammonia (80 ml) for 0.5 h. The resulting solid (ca.0.64 g) was removed by filtration, dried in vacuo. A portion (0.35 g) was stirred with methanesulfonic acid (0,10 g) in methanol (3.5 ml) and the mixture diluted with ether to give the mesylate of the title compound (0.14 g) as a white crystalline solid, of no sharp mp (deconip. > 270 C ii (500 MHz, dmso-d6) 2.31 (1.5H, s, SCH3), 5.42 (2H, brs, NCH2), 7.39 (5H, ni, aromatic H), 7.56 (2H, m, aromatic H), 7.86 (IH, dd, I = 7.2, 2. 4 Hz, aromatic H), 8.28 (lH, s, NH, exchang.), 8.4-8.8 (2H, vbrpeak, NH2, exchang.), 9.28 (IH, s, NH, exchang.).

The spectrum indicates stoichionietry of heterocycle.0.5 MeSO3H ni/z 347 (M+ 1).

Example 10

5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2(prop-2-enyl)-I,2,4-triazine hemi-methanesulfonate Allyl bromide (1.8 ml, 2.52 g, 0.02 mol) was added to a stirred suspension of 3,5-diamino-6-(2,3-dichlorophenyl)-I,2,4-triazine (1.28 g, 0.005 mo!) in acetone (50 ml). The mixture was stirred and heated at rellux for 4 days. More ally! bromide (0.6 ml, 0.007 mol) was added and refluxing continued for I day. After cooling, the solid was collected by filtration and then stirred with 0.88 aqueous ammonia (80 ml) for 0.5 h. The resulting solid (ca.0.84 g) was removed by filtration, dried in vacuo. A portion (0.30 g) was stirred with methanesulfonic acid (0.10 g) in methanol (3.5 ml) and the mixture diluted with ether to give the mesylate of the title conipound(0.26 g) as a fawn crystalline solid, of no sharp mp (decomp. > 270 C.

H (500 MHz, dmso-d6) 2.31 (l.5H, s, SCH), 4.74 (2H, d, J 4.8 Hz, NCH2), 5.28 (2H, in, olefinic H), 5.93 (IH, ni, olefinic H), 7.54 (2H, m, aromatic H), 7.86 (IH, m, aromatic H), 8.20 (IH, s, NH, exchang.), 8.2-8. 8 (2H, vbrpeak, NH2, exchang.), 9.2 (lH, s, NH, exchang.).

The spectrum indicates stoichiometry of heterocycle.0.5 MeSO3H.

Example II

5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-di hydro-3(5)-imino-1,2,4-triazin-2-ylJacetamide 2-(carboxamido)methyllamotrigine) 2-lodoacetamide (1.85 g, 0.01 mol) was added to a stirred suspension of 3,5-diamino-6-(2,3-dichloropheny!)-I,2,4-triazine (1.28 g, 0.005 mol) in acetone (50 ml). The mixture was stirred and heated at reflux for 4h. AfTer cooling, the solid was collected by filtration and then stirred with 0.88 aqueous ammonia (50 ml) for 0.5 h. The resulting solid (ca.l.1 g) was removed by filtration, dried in vacuo and recrystallised froni acelonitrile to give 0.56 g (36 %) of'the title compound as a white ctystalline solid of no sharp mp (decomp. above 270 C).

(500 MHz, drnso-d6) 2.07 (CHCN), 4.44 (2H, brs, NCH), 6.0-7.0 (2H, vbrpeak, NH2), 7.13 (1H, brs, NH, exchang.), 7.37 (IH, dd,J= 8,2 Hz, aromatic H), 7.44 (IH, t, J = 8 Hz, aromatic H), 7.49 (IH, brs, NH, exchang.), 7.71 (IH, dd, J = 8,2 Hz, aromatic H).

m/L313 (M)

Example 12

5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-1m1no2(4..

methyl)phenylmethyl-1,2,4-triazine 4-Methybenzyl bromide (0.70 g, 4.3 mmol) was added to a stirred suspension of 3,5-diarnino-6-(2,3-dichloropheny!)-1,2,4-triazjne (0.64 g, 2.5 mmol), Nal (0.1 g) and acetone (25 nil). The mixture was stirred and heated at reflux for 3 h. After cooling, the solid was collected by filtration and then stirred with 0.88 aqueous ammonia-water (40 ml, 1:1) for 0.5 h. The resulting solid (ca.0.7 g) was removed by filtration and dried in vacuo. Recrystallisation from ethanol gave the product (0.44 g) as a white solid, mpl8O-185 C (deconip.).

(500 MHz, dmso-d6) 2.27 (3H, s, CH3), 5.05 (2H, s, NCH,), 7. 14 (2H, d, J = 8 Hz, aromatic H), 7.22 (2H, d,J= 8Hz, aromatic H), 7.38 (IH, dd,J 7.5,2Hz, aromatic H), 7.44 (IH, d,.J= 7.5 Hz, aromatic H), 7.71 (1H, dd, I 7.5, 2 Hz). The spectrum indicates that the compound contains 0.3EtOH.

m/z 361 (M+ 1).

Example 13

5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imjno-2-(2,3..

difluoro)phenylmethyl-I,2,4-triazine 2,3-Difluorobenzyl bromide (2.07 g, 10 mmol) was added to a stirred suspension of 3,5-diamino-6-(2,3-dich!orophenyl)-1,2,4-triazine (1.28 g, 5 mrnol), Na! (0.! g) and acetone (50 nil). The mixture was stirred and heated at rellux for 6 h. After cooling, the solid was collected by filtration and then stirred with 0.88 aqueous ammonia-water (80 ml, 1:1) for 0.5 h. The resulting solid (ca.2 g) was renioved by filtration and dried in vacuo. Recrystallisation from methanol gave the product (1.2 g) as a pale yellow solid, nip 208-209 C.

ö (500 MHz, dmso-do) 5.20 (2H, s, NCH7), 5.66 (IH, brpeak, NH), 6.63 (lH, brpeak, NH), 7.19 (2H, ni, aromatic H), 7.35 (1H, m, aromatic H), 7.44 (2H, m, aromatic H), 7.72 (1H, brd,J= 7Hz, aromatic H).

nilz 383 (M+ 1)

Example 14

5(3)-Amino-6-(2.,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2(2-fluoro)phenylmethyl-1,2,4-triazine 2-Fluorobenzyl chloride (1.45 g, 10 mmol) was added to a stirred suspension of 3,5-diamino-6-(2,3-dichlorophenyl)-1,2,4-triazine (1.28 g, 5 mmol), NaT (0.1 g) and acetone (50 nil). The mixture was stirred and heated at reflux for 5 h. After cooling, the solid was collected by filtration and then stirred with 0.88 aqueous ammonia-water (80 ml, 1:1) for 0.5 h. The resulting solid (ca.2 g) was removed by filtration and dried in vacuo. Recrystallisation from methanol gave the product (1.2 g) as a pale yellow solid, mp 20 1-203 C.

H (500 MHz, dmso-d) 5.16 (2H, s, NCH2), 6-7 (2H, vbr peak, NH2), 7.18 (2H, rn, aromatic H), 7.33 (2H, m, aromatic H), 7.42 (2H, rn, aromatic H), 7.72 (1H, dd, J 7.5, 1.5 Hz). The spectrum indicated the compound contained 0.5McOH.

mlz 365 (M + I)

Example 15

5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2(3-fluoro)phenylmethyl-I,2,4-triazine 3-Fluorobenzyl chloride (1.45 g, 10 rnrnol) was added to a stirred suspension of 3,5-diarnino-6-(2,3-dichlorophenyl)-I,2,4-triazine (1.28 g, 5 mmol), Nal (0. I g) and acetone (50 ml). The mixture was stirred and heated at reflux for 5 h. After cooling, the solid was collected by filtration and then stirred with 0.88 aqueous ammonia-water (80 ml, 1: 1) for 0.5 h. The resulting solid (ca. I.5 g) was removed by filtration and dried in vacuo. Recryslallisation from methanol gave the product (0.42 g) as a pale yellow solid, nip 189-190 C.

(500 MHz, drnso-d6) 5.13 (2H, s, NCH7), 6-7 (2H, vbr peak, NH), 7.14 (3H, m, aromatic H), 7.43 (3H, m, aromatic H), 7.72 (IH, dd,J= 7.5, 1.5 Hz).

nilz 365 (M+ 1).

Example 16

5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2(4-fluoro)phenylmethyl-1,2,4-triazine 4-Fluorobenzy! chloride (1.45 g, 10 mrnol) was added to a stirred suspension of 3,5-diarnino-6-(2,3-dichlorophenyl)-l,2,4-triazine (1.28 g, 5 mmol), Nal (0.1 g) and acetone (50 nil). The mixture was stirred and heated at reflux for 5 h. After cooling, the solid was collected by liltration and then stirred with 0.88 aqueous ammonia-water (80 ml, 1: 1) for 0.5 h. The resulting solid (ca. 1.6 g) was removed by filtration and dried in vacuo. Recrystallisation from methanol gave the product (I. I g) as a pale yellow solid, mp 189-190 C.

(500 MHz, dmso-d6) 5.08 (2H, s, NCH2), 6-7 (2H, vbr peak, , NH, ), 7.17 (2H, t, J = 8 Hz, aromatic H), 7.40 (4H, m, aromatic H), 7.71 (lH, dd, J = 7, 2 Hz, aromatic H). rn/z 365 (M + I)

Example 17

5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-di hydro-3(5)-imino-2-(2-methoxy)phenylmethyl-I,2,4-triaiine 2-Methoxybenzyl chloride (1.56 g, 10 mrnol) was added to a stirred suspension of 3,5-diamino-6-(2,3-dichlorophenyl)-I,2,4-triazine (1.28 g, 5 mmo 1), Nal (0.1 g) and acetone (50 ml). The mixture was stirred and heated at reflux for 5 h. After cooling, the solid was collected by filtration and then stirred with 0.88 aqueous ammonia-water (80 ml, 1:1) fbr 0.5 h. The resulting solid (ca.1.8 g) was removed by filtration and dried in vacuo. Recrystallisation from ethanol gave the product (0.95 g) as a pale yellow solid, mp 194-196 C.

(500 MHz, dmso-d6) 3.80 (3H, s, OCH3), 5.05 (2H, brs, NCH2), 6.5-7.0 (IH, vbrpeak, NH), 6.92 (1 H, t, J = 8 Hz, aromatic H), 7.01 (2H, brt, J = 8 Hz, aromatic H), 7.26 (JH, brt,J= 8Hz, aromatic H), 7.40 (2H, ni, aromatic H), 7.69 (1H, brd,J 8 Hz, aromatic H).

m/z377(M+ 1)

Example 18

5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2(3-methoxy)phenylmethyl-I,2,4-triazine 3-Methoxybenzyl chloride (1.56 g, 10 mmol) was added to a stirred suspension of 3,5-dianiino-6-(2,3-dichlorophenyl)-l,2,4-triazine (1.28 g, 5 nmiol), Nal (0.1 g) and acetone (50 ml). The mixture was stirred and heated at rellux for 5 h. After cooling, the solid was collected by filtration and then stirred with 0.88 aqueous ammonia-water (80 ml, 1:1) for 0.5 h. The resulting solid (ca.1.4 g) was removed by filtration and dried in vacuo. Recrystallisation from ethanol gave the product (0.64 g) as a pale yellow solid, mp 192-195 C.

(500 MHz, dmso-d) 3.73 (3H, s, OCH3), 5.07 (2H, brs, NCH2), 6.5-7.0 (IH, vbrpcak, NH), 6.84 (1 H, brd, I = 8 Hz, aromatic H), 6.88 (2H, m, aromatic H), 7.26 (lH, 1, 1 = 8 Hz, aromatic H), 7.42 (2H, m, aromaticH), 7.71 (IH, d, I = 7 Hz,, aromatic H).

m/z377(M+ 1)

Example 19

5(3)-Amin o-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2-(4-methoxy) phenylmethyl-1,2,4-triazine 4-Methoxybenzyl chloride (1.56 g, 10 nimol) was added to a stirred suspension of 3,5-dianiino-6-(2,3-dichlorophenyl)-I,2,4-triazine (1.28 g, 5 mmol), Nal (0.1 g) and acetone (50 ml). The mixture was stirred and heated at reflux for 5 h. After cooling, the solid was collected by filtration and then stirred with 0.88 aqueous ammonia-water (80 ml, 1: 1) for 0.5 h. The resulting solid (ca. 1.6 g) was removed by filtration and dried in vacuo. Recrystallisation from ethanol gave the product (0.83 g) as a pale yellow solid, mp 212-2 15 C.

nilz 377 (M+ I)

Example 20

5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2(3-chloro)phenylmethyl-I,2,4-triazine 3-Chlorobenzyl bromide (2.05 g, 10 mrnol) was added to a stirred suspension of 3,5-diamino-6-(2,3-dichlorophenyl)-1,2,4-triazine (1.28 g, 5 mrnol), NaT (0.1 g) and acetone (50 ml). The mixture was stirred and heated at reflux for 5 h. After cooling, the solid was collected by filtration and then stirred with 0.88 aqueous ammonia-water (80 ml, 1:1) for 0.5 h. The resulting solid (ca.1.2 g) was removed by filtration and dried in vacuo. Recrysiallisation from methanol gave the product (0.35 g) as a pale yellow solid, mp 178-180 C.

(500 MHz, dmso-d6) 5.11 (2H, brs, NCH2), 6.5-7.0 (IH, vbrpeak, NH), 7.28 (IH, brd,J 8Hz, aromatic H), 7.32-7.47 (5H, m, aromatic H), 7. 72 (IH, dd,J= 7,2Hz, aromatic H). The spectrum indicates the presence of 0.75MeOH.

rnlz 381,383 (M + I)

Example 21

5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2(4-chloro)phenylmethyl-1,2,4-triazine 4-Chlorobenzyl chloride (1.61 g, 10 rnmol) was added to a stirred suspension of 3,5-dianiino-6-(2,3-dichlorophenyl)-I,2,4-triazine (1.28 g, 5 mmol), Nal (0.1 g) and acetone (50 ml). The mixture was stirred and heated at reflux for 5 h. After cooling, the solid was collected by filtration and then stirred with 0.88 aqueous ammonia-water (80 ml, 1:1) for 0.5 h. The resulting solid (ca.1.5 g) was removed by filtration and dried in vacuo. Recrystallisation from methanol gave the product (0.71 g) as a pale yellow solid, nip 192-193 C.

(500 MHL, dmso-d(,) 5.09 (2H, s, NCH), 5.5 (IH, vbrpeak, NH), 6.5 (IH, vbrpeak, NH), 7.35 (2H, d, J = 8 Hz, aromatic H), 7.42 (4H, rn, aromatic H), 7.72 (1 H, brd, I = 7 Hz, aromatic H).

m1z381,383(M+ 1)

Example 22

5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2(2-chloro)phenylmethyl-I,2,4-triazine 2-Chlorobenzyl bromide (2.06 g, 10 nimol) was added to a stirred suspension of 3,5-diamino-6-(2,3-dichlorophenyl)-l,2,4-triazine (1.28 g, 5 mrnol), Na! (0.1 g) and acetone (50 ml). The mixture was stirred and heated at reflux for 5 h. After cooling, the solid was collected by filtration and then stirred with 0.88 aqueous ammonia-water (80 ml, 1:1) for 0.5 h. The resulting solid (ca.1.5 g) was removed by filtration and dried in vacuo. Recrystallisation from methanol gave the product (0.71 g) as a pale yellow solid, rnp 205 C (decomp.).

H (500 MHz, dmso-d6) 3.32 (3H, s, MeOH), 5.18 (2H, s, NCH2), 5.6 (IH, brpeak, NH, exchang.), 6.6 (1H, brpeak, NH, exchang.), 7.20 (lH, m, aromatic H), 7.32 (2H, m, aromatic H), 7.46 (3H, m, aroniatic H), 7.70 (1H, brd,1 7Hz, aromatic H).

m/z38l,383(M+ 1)

Example 23

5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2(2-trifluoromethyl)phenylmethyl-1,2,4-triazine 2-trifluoromethylbenzyl bromide (0.56 g, 2.2 mrnol) was added to a stirred suspension of 3,5-diamino-6-(2,3-dichlorophenyl)-I,2,4-triazine (0.56 g, 2.2 mmo 1), Na! (0.1 g) and acetone (25 ml). The mixture was stirred and heated at reflux for 5 h. After cooling, the solid was collected by filtration and then stirred with 0.88 aqueous ammonia-water (40 ml, 1: 1) for 0.5 h. The resulting solid (ca. 1.5 g) was removed by filtration and dried in vacuo. Recrystallisation from methanol gave the product (0.42 g) as a pale yellow solid, mp 200-201 C.

H (500 MHz, dmso-d6) 3.32 (3H, s, MeOH), 5.31 (2H, s, NCH2), 5.66 (IH, brs, NH, exchang.), 6.66 (1H, brs, NH, exchang.), 7.31 (1H, d, J = 8 Hz, aromatic H), 7.45 (3H, m, aromatic H), 7.5 (IH, br peak, NH, exchang.), 7.72 (3H, m, aromatic H).

m/z4l4,416(M+ 1)

Example 24

5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2(3-trifluoromethyl)phenylmethyl-I,2,4-triazine 3-Trifluorornethylbenzyl bromide (2.0 g, 8.7 mrnol) was added to a stirred suspension of 3,5-diamino-6-(2,3-dichlorophenyl)-l,2,4-triazine (1,12 g, 4.4 mmol), Nat (0.1 g) and acetone (50 nil). The mixture was stirred and heated at reflux for 5 h. After cooling, the solid was collected by filtration and then stirred with 0.88 aqueous ammonia-water (80 ml, 1:1) for 0.5 h. The resulting solid (ca. 1.2 g) was removed by filtration and dried in vacuo. Recrystallisation from methanol gave the product (0.52 g) as a pale yellow solid, mp 168-170 C (500 MHz, dmso-d6) 3.32 (3H, s, MeOH), 5.22 (2H, brs, NCH2), 5.4-5.8 (1H, vbrpeak, NH, exchang.), 6.4-6.8 (IH, brs, NH, exchang.), 7.42 (2H, m, aromatic H), 7.63 (4H, ni, aromatic H), 7.73 (1 H, brd, J = 7 Hz, aromatic H). mlz 414, 416 (M + 1).

Example 25

5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2(4-trifluoromethyl)phenylmethyl-I,2,4-triazine 4-Trifluoromethylbenzyl chloride (1.0 g, 4.0 mrnol) was added to a stirred suspension of 3,5-diarnino-6-(2,3-dichlorophenyl)-l,2,4-triazine (0.56 g, 2.2 nimol), Nal (0.1 g) and acetone (25 nil). The mixture was stirred and heated at reulux for 5 h. After cooling, the solid was collected by filtration and then stirred with 0.88 aqueous ammonia-water (40 nil, I: I) for 0.5 h. The resulting solid (ca.0.7 g) was removed by filtration and dried in vacuo. Recrystallisation froni methanol gave the product (0.42 g) as a pale yellow solid, mp 198-200 C.

H (500 MHz, dniso-d6) 3.32 (3H, s, MeOH), 5.20 (2H, brs, NCH2), 5.3-5.8 (IH, vbrpeak, NH, exchang.), 6.4-6.8 (IH, brs, NH, exchang.), 7-8 (lH, vbrpeak, NH, exchang.), 7.43 (2H, m, aromatic H), 7.53 (2H, brd, J = 8 Hz, aroniatic H), 7.73 (3H, brd,J 8Hz, aromatic H). ni/z 414, 416 (M + I)

Example 26

5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2(2-fluoro-3-trifluoromethyl)phenylmethyl-I,2,4-triazine 2-Fluoro-3-trifluorornethylbenzyl bromide (1.0 g, 4.0 mmol) was added to a stirred suspension of 3,5-diarnino-6-(2,3-dichlorophenyl)-I,2,4-triazine (0.56 g, 2.2 mmo I), Nal (50 mg) and acetone (25 ml). The mixture was stirred and heated at reflux for 5 h. After cooling, the solid was collected by filtration and then stirred with 0.88 aqueous ammonia-water (40 nil, 1: 1) for 0.5 h. The resulting solid (ca.0.7 g) was renioved by filtration and dried in vacuo. Recrystallisation from methanol gave the product (0.40 g) as a pale yellow solid, mp> 250 C (decomp.).

ö (500 MHz, dmso-d(,) 3.32 (3H, s, McOH), 5.23 (2H, brs, NCH2), 5.67 (IH, brs, NH, exchang.), 6.5-7.0 (1H, vbrpeak, NH, exchang.), 7.42 (3H, m, aromatic H), 7.5 (IH, brpeak, NH, exchang.), 7.70 (3H, m, aromatic H).

m/z432,434(M+l)

Example 27

4-I5(3)-Amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3(5)-imino-1,2, 4-triazin-2-yl] methyl}benzamide or 5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2(3-carboxamido)phenylmethyl-1,2,4-triazine 3-(chloromethyl)benzami de was prepared by reaction of 3 -ch loromethylbenzoyl chloride with ammonia according to the procedure of C. Y. Watson et al, Bioorg. & Med (hem., 6,721-734(1998).

(3-Chloromethyl)benzamide (1.33 g, 7.8 rnmol) was added to a stirred suspension of 3,5-diarnino-6-(2,3-dichlorophenyl)-I,2,4-triazine (1.61 g, 6.3 mmo I), NaT (0.1 g) and acetone (70 ml). The mixture was stirred and heated at reflux overnight. After cooling, the solid was collected by filtration and then stirred with 0.88 aqueous ammonia-water (80 nil, I I) for 0.5 h. The resulting solid (ca. 1.5 g) was removed by filtration and dried in vacuo. Recrystallisation from methanol gave the product (0.92 g) as a pale yellow solid, nip 228-230 0(' j (500 MHz, dmso-d) 3.32 (3H, s, MeOH), 5.15 (2H, brs, NCH), 5.4-5.8 (1H, vbrpcak, NH, exchang.), 6.4-6.8 (IH, vbrpcak, NH, exchang.), 7.34 (1H, brs, NH, exchang.), 7.38-7.48 (4H, rn, aromatic H), 7.70 (IH, brd,J= 8Hz, aromatic H.), 7.76 (IH, brs, I = 8 Hz, aromatic H), 7.84 (1H, brs, aromatic H), 7.96 (1H, brs, NH, cxchang.). nh/z 389, 391 (M + 1).

Example 28

4-fl5(3)-Amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3(5)-imino-1,2, 4-triazin-2-yll methyl}phenylmethanol or 5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2(4-hydroxymethyl)phenylmethyl-I,2,4-triazine 4-(Chlorornethyl)benzyl alcohol (1.0 g, 6.4 mniol) was added to a stirred suspension of 3,5-diamino-6-(2,3-dichlorophenyl)-1,2,4-triazine (1.28 g, 5 niniol), Nal (0.1 g) and acetone (50 ml). The mixture was stirred and heated at reflux for 10 h. After cooling, the solid was collected by filtration and then stirred with 0.88 aqueous amnionia-water (80 ml, 1:1) for 0.5 h. The resulting solid (Ca. 1.3 g) was removed by filtration and dried in vacuo. Recrystallisation from methanol gave the product (0.47 g) as a pale yellow solid, rnp 215-217 C.

(500 MHz, dmso-d6) 3.32 (3H, s, MeOH), 4.47 (2H, d, I = 5 Hz, OH, exchang.), 5.08 (2H, brs, NCH2), 5.14 (IH, brt,J= 5Hz, OH, cxchang.), 5.4-5.8 (IH, vbrpeak, NH, exchang.), 6.4-6.8 (1H, vbrpeak, NH, exchang.), 7.28 (4H, m, aromatic H), 7.40 (I H, brd, I = 8 Hz, aromatic H), 7.45 (IH, t, J = 8 Hz, aromatic H), 7.72 (1H, brd, I = 8 Hz, aromatic H).

mIz376,378(M+ I)

Example 29

5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2(3-thienylmethyl)-l,2,4-triazine 3-chlorornethyithiophene was prepared by chlorination of thiophene-3-methanol according to the procedure of S. Gronowitz and S. Liljefors, chetnica Scipta, 13, 39-(1978-79).

2-Chloromethyithiophene (1.04 g, 7.8 mmol) was added to a stirred suspension of 3,5-diamino-6-(2,3-dichlorophenyl)-I,2,4-triazine (1.00 g, 3.9 nirnol), Na! (0.07 g) and acetone (35 ml). The mixture was stirred and heated at rellux for 36 h. After cooling, the solid was collected by filtration and then stirred with 0.88 aqueous ammonia-waler (30 ml, I: I) for 5 h. The resulting solid (ca.0.5 g) was removed by filtration and dried in vacuo. Recrystallisation from niethanol gave the product (0.22) as a cream solid, nipl9I-192 C (decomp.).

(500 MHL, dmso-d6) 3.32 (3H, s, CHOH), 5.07 (2H, s, NCH2), 5.2-6.0 (IH, vbr peak, NH, exchang.), 6.5-7.5 (2H, vbrpeak, NH), 7.11 (IH, dd, 1=5, 1 Hz, aromatic H), 7.37-7.45 (3H, rn, aromatic H), 7.50 (IH, m, aromatic H), 7.72 (1H, dd,J 7.5,2 Hz). The spectrum indicates that the compound contains 1.OMeOH. m/z 353 (M + 1).

Example 30

5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imi no-2-(3-furanylmethyl)-1,2,4-triazine 3-chloromethylfuran was prepared by chlorination of furan-3-methanol according to the procedure of E. Sherman and E. D. Amstutz, 1. Am. chem. Soc., 72, 2195-2199 (1950).

3-Chloromethylfuran (0.90 g, 7.7 rnrnol) was added to a stirred suspension of 3,5-diarnino-6-(2,3-dichlorophenyl)-I,2,4-triazine (1.00 g, 3.9 mmol), Nal (0.07 g) and acetone (40 nil). The mixture was stirred and heated at reflux for 36 h. After cooling, the solid was collected by filtration and then stirred with 0.88 aqueous ammonia-water (30 ml, I: I) for 5 h. The resulting solid (ca. 1.1 g) was removed by filtration and dried in vacuo. Recrystallisation from methanol gave the product (0.72 g) as a cream solid, nipl9I-193 C.

H (500 MHz, dmso-d6) 3.32 (3H, s, CH3OH), 4.92 (2H, s, NCH2), 5.5-6.4 (IH, vbr peak, NH, exchang.), 6.48 (IH, brs, furan H), 6.5-7.5 (2H, vbrpeak, NH,), 7.37-7.46 (2H, m, aromatic H), 7.61 (IH, brs, furan H), 7.64 (JH, brs, furan H), 7.71 (1H, dd, I = 7.5, 2 Hz, aromatic H). The spectrum indicates that the compound contains 1.OMeOH.

m/z337(M+ 1)

Example 31

6-(2,3,5-trichlorophenyl)-I,2,4-triazine-3,5-diamine prepared according to the method described in US patent 4,602,017: rnp 232-235 C

Example 32

5(3)-Amino-6-(2,3,5-trichlorophenyl)-2,3(2,5)-dihydro-3(5) -imino-2-methyl-1,2,4-triazine lodornethane (0.40 g, 2.8 mmol) was added to a stirred suspension of 3,5-diamino-6-(2,3,5-trichlorophenyl)-I,2,4-triazine (0.20 g, 0.7 mmo I) in acetone (15 nil). The mixture was stirred at room temperature for 6 days and the solvent removed in vacuo at 40 C. ice (ca. 4 g) was added to the residue followed by 0.88 aqueous ammonia (3 ml) and the mixture stirred for 4 h. The solid was removed by filtration, dried in vacuo and recrystalllised from ethanol to give 0.13 g of the title compound as an off-white crystalline solid, mp 225-226 C.

(500 MHz, dmso-d(,) 3.47 (3H, s, NCH3), 5.5-7.4 (3H, vbr peak, NH, exchang.), 7.56 (1H, d,J= 2.5 Hz, aromatic H), 7.92 (IH, dd,J= 2.5 Hz, aromatic H).

mIz304-306(M+ I)

Example 33

6-(2,3-ditluorophenyl)-I,2,4-triazine-3,5-diamine Stepi 2,3-Difluorobenzoyl chloride 2,3-Difiuorobenzoic acid (11.6 g, 0.07 niol), thionyl chloride (37.5 ml, 61.1 g, 0.5 mol) and toluene (80 nil) were heated at reflux for 3 h. The solution was cooled and the volatiles were renioved in vacuo. The residue was azeotroped with toluene (2 x 30 nil) to give the product (10.8 g) as a clear yellow oil.

Step 2 2,3-Difluorobenzoyl cyanide Copper(l) cyanide (6.6 g, 0.07 mol), potassium iodide (12.2 g, 0.07 mol) and xylene (70 ml) were heated at reflux for 24 h using a Dean-stark apparatus. A solution of 2,3-difluorobenzoyl chloride (10.8 g, 0.06 mol) in xylene (40 nil) was added. The resulting suspension was refluxed under N2 at 165 C for 3 days using a Dean-Stark apparatus. After cooling, inorganic salts were removed by filtration and the filtrate concentrated in vacuo. The residue was azeotroped with toluene (2 x 30 ml) to give the product (7.2 g) as a brown solid.

Step 3 2-(2,3-Difluorophenyl)-2-guanidinoimino)acetonitrile Cone. sulfuric acid (43.5 nil, 80 g, 0.82 mol) was added slowly to water (45 ml) with stirring. Aminoguanidine bicarbonate (4.4 g, 0.032 mol) was added slowly to this hot acid solution with stirring (caution! CO2 evolved) and stirring was continued for a further 15 mm. A solution of 2,3-difluoro-benzoyl cyanide (3.1 g, 0.0 19 mol) in acetonitrile (20 ml) was added dropwise over 0.5h to the above solution of aminoguinidine sulfate and the mixture was stirred at room temperature for 4 days.

Aqueous NaOH solution (4M) was then added carefully, with cooling in an ice-bath, until the mixture was at pH 7. The precipitate was collected by filtration, washed with water and dried to give the product (2.9 g) as a yellow solid, mp 168-170 C. Step4

2-(2,3-Diuluorophenyl)-2-guanidinoimino)acctonitrile (2.8 g, 0.01 mol) and propan-I -ol (30 ml) were stirred and heated at rellux for 1.5 h. The cool solution was concentrated in vacuo and the residue chrornatographed on silica (250 g). Elution with CH2CI2-MeOH (95:5) gave a tan solid. This material was slurried in CH2CI2 and the remaining insolubles collected by filtration to give the product (1.3 g) as a cream solid, rnp 229-230 C.

8 (500 MHz, dmso-d6) 6.42 (2H, brs, NH, exchang.), 6.6-7.0 (2H, vbr peak, NH2, exehang.), 7.25 (IH, brt,J= 7.5 Hz, aromatic H), 7.30 (IH, m, aromatic H), 7.48 (1H, m, aromatic H). m/z 224 (M + I)

Example 34

5(3)-Amino-6-(2,3,5-trichlorophenyl)-2,3(2,5)-dihydro-3(5) -imino-2-propyl-1,2,4-triazine lodopropane (0.51 g, 3 mrnol) was added to a stirred suspension of 3,5-diamino-6-(2,3,5-trichlorophenyl)-I,2,4-triazine (0.29 g, 1 mmol) in acetone (15 ml). The mixture was stirred and heated at reflux for 8 days. After cooling, the precipitate was collected by filtration, and then stirred in water ( 4 ml) and aqueous ammonia (2 ml) for 4 h. The solid was removed by filtration, dried in vacuo and recrystalllised from niethanol to give 0.15 g of the title compound as an off-white crystalline solid, nip 240-243 C.

(500 MHz, dniso-d6) 0.88 (3H, 1, J = 7.5 HL, CH), 1.67 (2H, hext, I = 7.5 Hz, CH2), 3.82 (2H, t, J = 7.5 Hz, NCH2), 6.3-7.3 (2H, vbr peak, NH2, exchang.), 7.56 (IH, d,J= 2.5 Hz, aromatic H), 7.92 (lH, d,J= 2.5 Hz, aromatic H).

rn/i 332-334 (M + 1)

Example 35

3-chloro-2-fluorophenyl)-1,2,4-triazine-3,5-diamine Step 1 3-Chloro-2-fluorobenzoic acid 3-Chloro-2-Iluorohenzaldehyde (15.9 g, 0.1 mol) was dissolved in tert-butanol (60 ml), stirred and heated under N-at 50 C. 2M aqueous NaOH (100 nil, 0.2 mol) was warmed to 50 C and added to the solution of the aldehyde. Aqueous hydrogen peroxide solution (H207, 30%, 70 ml, 0.6 mol) was added over 45 mm, niaintaining the temperature at 55-60 C. The mixture was stirred and heated under N2 for a further lh, cooled and concentrated in vacuo. The residual slurry was filtered. The filtrate was washed with toluene (2 x xx ml) and acidified to pH 1 with 5N hydrochloric acid whilst stirring vigorously. The resulting solid was collected by filtration, washed with water and dried in vacuo at 50 C. to give 11.1 g of product, nip 179-181 C.

A sample prepared by a different route [ J.Mortier ci al, Tetrahedron Lett., 36, 881- 884 (1995)] is reported to have nip 179-18 1 C.

Step 2 3-Chloro-2-fluorobenzoyl chloride 3-Chloro-2-lluorobenzoic acid (10.0 g, 0.06 mol), thionyl chloride (31 ml, 50 g, 0.4 mol) and dry toluene (40 nil) were heated at reflux for 3 h. The solution was cooled and the volatiles were removed in vacuo. The residue was azeotroped with toluene (2 x 30 ml) to give the product (11.5 g) as a clear yellow oil.

Step 3 3-Chloro-2-\fluorobenzoyl cyanide Copper(1) cyanide (6.6 g, 0.07 mol), potassium iodide (12.2 g, 0.07 mol) and xylene (50 ml) were heated at reflux for 24 h using a Dean-stark apparatus. A solution of 3-chloro-2-fluorobenzoy! chloride (11.5 g, 0.06 mol) in xylene (15 ml) was added. The resulting suspension was refluxed under N2 at 165 C lbr 3 days using a Dean-Stark apparatus. After cooling, inorganic salts were removed by liltration and the filtrate concentrated in vacuo. The residue was azeotroped with toluene (2 x 30 nil) to give the product (9.5 g) as a brown solid.

Step 4 2-(3-Chloro-2-fluorophenyl)-2-guanidinoimino)acetonitrile Cone. sulfurie acid (43.5 ml, 150g. 1.6 mol) was added slowly to water (45 ml) with stirring. Aniinoguanidine bicarbonate (5.7 g, 0.036 mol) was added slowly to this hot acid solution with stirring (caution! CO2 evolved) and stirring was continued for a further 15 mm. A solution of 3-chloro-2-tluoro-benzoyl cyanide (4.3 g. 0.02 mo!) in acetonitrile (31 nil) was added dropwise over 0.5h to the above solution of aniinoguinidine sulfate and the mixture was stirred at room temperature for 4 days.

Aqueous NaOH solution (4M) was then added carefully, with cooling in an ice-bath, until the mixture was at pH 7. The precipitate was collected by filtration, washed with water and dried to give the product (3.2 g) as a tan solid.

Step 5 2-(3-Chloro-2-fluorophenyl)-2-guanidinoirnino)acetonitrile (3.2 g, 0.01 mol) and propan-1-ol (30 ml) were stirred and heated at reulux for 3 h. The cool solution was concentrated in vacuo and the residue chrornatographed on silica (250 g). Elution with CH2CI2-MeOH (95:5) gave a tan solid. This material was slurried in CH2CI2 and the remaining insolubles collected by filtration to give the product (1.3 g) as a creani solid, nip 246-247 ö (500 MHz, dmso-d6) 6.99 (2H, brs, NH7, exchang.), 7.28 (1 H, t, .1 = 8 Hz, aromatic H), 7.44 (I H, td, J = 8, 2 Hz, aromatic H), 7.65 (1 H, td, J = 8, 2 Hz, aromatic H), 12.5 (1H, brpeak, NH, exchang.).

niIz240,242(M+ 1).

Example 36

5(3)-Amino-6-(3-chloro-2-fluorophenyl)-2,3(2,5)-dihydro-3(5) -imino-2-methyl-I,2,4-triazine lodomethane (0.5 ml, 1.14 g, 8 nirnol) was added to a stirred suspension of 3,5-diamino-6-(3-chloro-2-Iiuorophenyl)-I,2,4-triaine (0.48 g, 2 mmo I) in acetone (25 ml). The mixture was stirred at 45 C for 24h, cooled, and the solid collected by filtration. Ice (ca. 10 g) was added to the residue followed by 0.88 aqueous ammonia (5 ml) and the nhixture stirred for 4 h. The solid was renioved by filtration, dried in vacuo and recrystalllised from methanol to give 0.23 g of the title compound as an off-white crystalline solid, mp 194-196 C.

H (500 MHz, dmso-d() 3.48 (3H, s, NCH3), 6.2-7.2 (2H, vbr peak, NH2, exchang.), 7.27 (lH, t,J= 8Hz, aromatic H), 7.40 (1H, td,J= 8,2Hz, aromatic H), 7.64 (1H, td, 18,2 Hz, aromatic H).

mIz254,256(M+ 1)

Example 37

1,1 -Dimethylethyl 4-15(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-I,2, 4-triazin-2-ylmethyl J piperidine-I -carboxylate or 5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2(Bocpiperidin- 4-yl)methyl-1,2,4-triazine 1,1 -Dirnethylethyl 4-iodornethylpiperidine-I -carboxylate [prepared from ethyl isonipecotate by a three step process according to the method of A. Villalobos et a!, 1. Mcd. chem., 37, 272 1-2734 (1994)].

(3.25 g, 10 mmol) was added to a stirred suspension of 3,5-diarnino-6-(2,3-dichlorophenyl)-l,2,4-triazine (1.28 g, 5 rnrnol) in acetone (50 ml). The mixture was stirred and heated at reflux for 9 days. After cooling in ice, the solid (2.5 g) was collected by filtration. This material was stirred in water (10 ml) and 0.88 aqueous ammonia (10 ml) for 12 h. The solid was renioved by filtration, dried in vacuo and recrystallised from methanol to give 0.60 g of the above urethane as an off-white crystalline solid, of no sharp mp.

m/z 453,455 (M + 1) oH (500 MHz, dn1so-d(,) 1.05 (2H, ddd, J = 25, 12, 4 Hz, CCHC), 1.38 (9H, s, C(CH3)3), 1.59 (2H, brd,J= 12Hz, CCHC), 2.06 (IH, iii, CH), 2.69 (2H, m, CH2N), 3.76 (2H, in, CH2N), 3.92 (2H, brd, J= 7 Hz, NNCH), 5.0-6.0 (1H, vbr peak, NH, exchang.), 6.4-7.0 (2H, vbrpeak, NH, cxchang.), 7.39 (1H, d, I = 7.5 Hz, aromatic H), 7.44 (IH, t, I = 7.5 Hz, aromatic H), 7.70 (IH, d, I = 7.5 Hz, aromatic H).

Example 38

4-15(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-1,2, 4-triazin-2-ylmethyljpiperidine dimethanesulfonate or 5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-i mino-2-(piperidin-4-yl)methyl-1,2,4-triazine 1,1 -Dimethylethyl 4-[5(3)-Aniino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-I,2, 4-triazin-2-ylmethyl]piperidine-I -carboxylate (0.5 g, 1. I mmol) was dissolved in CH2CI2 (10 nil) and trifluoroacetic acid (TFA, 10 ml) added. The mixture was stirred for 0.5h, then concentrated in vacuo. Remaining TFA was renioved by azeotroping with toluenc. The residue was stirred with saturated NaHCO3 solution (10 nil) and then ammonia (d = 0.88) was added until the pH was 12. The mixture was stirred 2h, the solid which deposited collected by filtration and dried. This material (0.12 g, 0.3 nimol) was dissolved in methanol (3 ml) and niethanesulfonic acid (70 mg, 0.7 rnmol) added. The solution was stirred for 2h and then diluted slowly with ether until an oily solid precipitated. This was triturated and removed by filtration and dried in vacuo to give the product (0.18 g) as a off-white solid, mp 180-200 C.

m/z 353,355 (M + 1).

OH (500 MHz, dniso-d6) 1.30 (2H, brddd,J= 25, 12,4Hz, CCH2C), 1.79 (2H, brd,J = 12Hz, CCHC), 2.07 (IH, m, CH), 1.59 (2H, brd,J 12Hz, CCH2C), 2.07 (IH, rn, CH), 2.31 (6H, rn, CH3S), 2.76 (2H, td, I = 12, 4 Hz, CH2N), 3.20 (2H, brd, I = 12 Hz, NCH2), 3.22-3.40 (7H, brpeak, NH, exchang.), 4.00 (2H, brd, I = 7 Hz, NNCH), 7.53 (1 H, dd, I = 7.5, 2 Hz, aromatic H), 7.56 (1 H, t, I = 7.5 Hz, aromatic H), 7.86 (IH, dd,I= 7.5,2Hz, aromatic H).

Example 39

5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2-(3,3, 3-trifluoropropyl)-I,2,4-triazine no sharp mp (decomp.) (500 MHz, dmso-d6) 2.76-2.86 (2H, rn, CH2CF3), 4.3 1 (2H, 1, 1 = 7 Hz, NCH2), 7.47 (IH, dd, 1=8, 1.5 HL, aromatic H), 7.53 (IH, t,J= 8 Hz, aromatic H), 7.84 (IH, dd, J = 8, 1.5 Hz, aromatic H), 8.30 (3H, brpeak, NH, exchang.). rnlz 352,354 (M + 1).

Example 40

2-chloro-3-tluorophenyl-I,2,4-triazine-3,5-diamine This compound was prepared in a similar manner to Example 35 via the following intermediates: Step I 3-Chloro-2-fluorobenzoic acid was obtained by chlorination of 3-fluorobenzoic acid according to the method of B. Bennetau et a!, 1. Ghetn. Soc. Perkin Trans 1, 1265- 1271 (1995).

Step 2 3-Chloro-2-fluorobenzoyl chloride Step 3 3-Chloro-2-fluorobenzoyl cyanide Step 4 2-(3-Chloro-2-fluorophenyl)-2-guanidinoimino)acetonitrile Step 5 2-chloro-3-lluorophenyl-1,2,4-triazine-3,5-diamine, mp 244-246 C, rn.lz 240, 242 (M+ 1).

(500 MHz, dmso-d) 6.4-7.0 (4H, s + brpeak, 2 x NH2, exchang.), 7.25 (IH, m, aromatic H), 7.47 (2H, 111, aromatic H).

Example 41

3,5-Diamino-6-(2,5-dichlorophenyl)-1,2,4-triazine, nip 228-230 C, was prepared according to the niethod described in US Patent No 4,602,017.

Example 42

3,5-Diamino-6-(3,5-dichlorophenyl)-I,2,4-triazine, mp 223-225 C, was prepared from 3,5-dichlorobenzoic acid using similar methodology to that used in Example 33.

Example 43

3,5-Diamino-6-phenyl-1,2,4-triazine, mp 218-219 C, was prepared using the method of J.A. Settepani and A. B. Borkovec,J. Iletemcvl. (hem., 3, 188-190, (1966)

Example 44

3,5-Diamino-6-(2,4-dichlorophenyl)-1,2,4-triazine was prepared according to the method of R.W.A. Rees and P.B. Russell et al,J.Med. Chem., 15, 859-861 (1972).

Example 45

5(3)-Amino-6-(2.,4-dichlorophenyl)-2,3(2,5)-dihydro-3(5) -imino-2-methyl-1,2,4- triazine methanesulfonate, mp 283-285 C, was prepared by reaction of 3,5-Diarnino-6-(2,4-dichlorophenyl)-1,2,4triazine and methyl methanesulfonate in a similar manner to Example 2, but using ethanol as solvent. This compound is described US Patent No. 4,649,139.

Example 46

5(3)-Am ino-6-phenyl-2,3(2,5)-dihydro-3(5)-imino-2-methyl-1,2,4-triazine methanesulfonate, nip 230-232 C, was prepared by reaction of 3,5-Diamino-6-phenyl-l,2,4-triazine and methyl rnethanesulfonate in a similar manner to that of example 2, but using ethanol as solvent. The free base is described in US pat 4,649,139.

Example 47

5(3)-Amino-6-phenyl-2,3(2,5)-dihydro-3(5)-i mino-2-ethyl-I,2,4-triazine methanesulfonate, rnp 230-232 C, was prepared by reaction of 3,5-Diarnino-6-phenyl-1,2,4-triazine and ethyl methanesulfonate in a similar manner to that of Example 2, but using ethanol as solvent.

Example 48

5(3)-Amino-6-(2,5-dichlorophenyl)-2,3(2,5)-dihydro-3(5) -imino-2-methyl-1,2,4-triazine methanesulfonate, rnp 297-298 C, was prepared according to the method described in US Patent No. 4,649,139.

Example 49

5(3)-Amino-6-(2,5-dichlorophenyl)-2,3(2,5)-dihydro-3(5) -imino-2-ethyl-1,2,4- triazine methanesulfonate, mp 264-265 C, was prepared by reaction of 3,5-Diamino-6-(2,5-dichlorophenyl)-I,2,4-triazine with ethyl methanesulfonate in a similar manner to that described in Example 2, but using ethanol as solvent.

Example 50

5(3)-Amino-6-(2,3,5-trichlorophenyl)-2,3(2,5)-dihydro-3(5) -imino-2-ethyl)-1,2,4- triazine mp 269-27 1 C, was prepared by reaction of 3,5-Diamino-6-(2,3,5-trichlorophenyl)-l,2,4-triazine with ethyl methansulfonate in a similar manner to that described in Example 2, but using ethanol as solvent.

Example 51

3,5-Diamino-6-(2-trifluoromethoxyphenyl)-I,2,4-triazine, mp 148-150 0C, was prepared from 2-trilluoroniethoxybenzoic acid using similar methodology to that employed for Example 33

Example 52

5(3)-Amino-6-(2,3-dichlorophenyl)-2,3(2.,5)-dihydro-3(5)-imino-2(2-fluoroethyl)- 1,2,4-triazine methanesulfonate Stepi 2-Fluoroethyl methanesulfonate Methanesulfonyl chloride (12.6 g, 0.11 mol) was added over 10 mm to a stirred solution of 2-fluoroethanol ( 6.40 g, 0.10 niol) in dichloromethane (100 ml) containing triethylaniine (12.1 g, 0.12 mol) kept at 0-5 C. The mixture was stirred for lh and allowed to warm to room temperature over this tinie. The mixture was diluted with dichloromethanc (25 ml) , and then washed with ice-water ( 40 nil), followed by cold 10% hydrochloric acid ( 40 ml), saturated sodium bicarbonate solution ( 40 ml) and brine (40 ml). The dichiorornethane solution was dried over sodium sulfate and the solvent removed in vacuo to give the product as a pale yellow oil (11.4 g). This material was used without further purification for the following reaction.

Step 2 2-fluoroethanol (0.50 g, 3.5 nirnol), 3,5-diamino-6-(2,3-dichlorophenyl) -1,2,4-triazine (0.50 g, 2.0 mmol) and dimethylformamide (4 ml) were stirred and heated at C for 24h. The solution was cooled, ether (30 ml) added, and the mixture stirred and triturated for 0.5h. After the mixture settled, the solvent was decanted from the oily precipitate and the residue extracted with boiling 2-butanone (25 nil, 2 x) to remove impurities. Crystallisation of the residue from methanol-ether gave the title conipound as a light tan solid (0.40 g), nip 253-255 C (deconip., rapid heating) öi (500 MHz, drnso-d6) 2.31 (3H, s, SCH3), 4,46 (2H, brdt, J = 26.4, 5Hz, NCH2), 4.76 (2H, brd, I = 47.2 Hz, FCH7), 7.55 (2H, m, aromatic H), 7.86 (I H, m, aromatic H), 8.28 (1H, s, NH, exchang.), 8.3-9.0 (2H, vbrpcak, NH2, exchang.), 9.24 (IH, s, NH, exchang.)

Example 53

5(3)-Amino-6-(3,5-dichlorophenyl)-2,3(2,5)-dihydro-3(5) -imino-2-methyl-1,2,4- triazine methanesulfonate, nip 234-236 C, was prepared by reaction of 3,5-Diarnino-6-(3,5-dichlorophenyl)-1,2,4triazinc and methyl methanesul fbnate in a similar manner to Example 2, but using ethanol as solvent.

Example 54

5(3)-Amino-6-(3,5-dichlorophenyl)-2,3(2,5)-dihydro-3(5) -imino-2-ethyl-1,2,4- triazine methanesulfonate, nip 217-219 C, was prepared by reaction of 3,5-Dianiino-6-(3,5-dichlorophenyl)-1,2,4triazine and ethyl niethanesulfonate in a similar manner to Example 2, but using ethanol as solvent

Example 55

5(3)-Amino-6-(2,3,5-trichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2(2-fluoroethyl)-1,2,4-triazine methanesulfonate, nip 212-214 C, was prepared by reaction of 2-Iluoroethyl methanesul fonate with 3,5-diamino-6-(2,3,5-trichlorophenyl) -1,2,4-triazine in dimcthylformaniide in a similar manner to that of Example 52.

Example 56

5(3)-Amino-6-(2,3,5-trichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2-(3, 3,3-trifluoropropyl)-1,2,4-triazine methanesulfonate Step I 3,3,3-Trilluoropropyl niethanesulfonate was prepared by reaction of 3,3,3-trifluoropropanol with methanesulionyl chloride by an analogous procedure to that used for Example 52 step 1.

Step 2 Reaction of 3,3,3-trilluoropropyl methanesulfonate with 3,5-diamino-6-(2,3,5-trichlorophcnyl) -I,2,4-triazine in dimethylformarnide in a similar manner to that of Example 52 gave the title conipound of no sharp mp (hygroscopic).

Example 57

5(3)-Amino-6-(2,3,dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2-(2, 2-difluoroethyl)-! ,2,4-triazine Step! 2,2-Difluoroethyl trifluoroniethanesulfonate was prepared by reaction of 2,2difluoroethano! and triflic anhydride according to the procedure of W. G. Reifenrath et al,J. Med. chein., 23, 985-990 (1980).

Step 2 2,2-Difluoroethyl trifluoromethanesulfonate (1.40 g, 6.5 mmol) was added to 3,5-diamino-6-(2,3-dichlorophenyl) -1,2,4-triazine (0.50 g, 2.0 mmol) and dimethylformamidc (3.5 ml). The warm mixture was stirred and heated at 100 C for 2h and then allowed to stand at room temperature overnight. Ether (35 ml) was added, and the mixture stirred for 0.Sh. After the mixture settled, the solvent was decanted from the oily precipitate and the residue stirred with water (10 nil) and aqueous ammonia solution (5 ml, d = 0.88) for 6h. A tan solid was removed by filtration, washed with water (3 nil) and air-dried. Recrystallisation from propan-2-ol gave the title compound as a light tan solid (0.25 g), nip 179-181 C (decomp., rapid heating) (500 MHz, dmso-d6) 4.30 (2H, brt, J = 13.8 Hz, NCH), 5.6-7.0 (2H, vbrpeak, NH, exchang.), 6.39 (IH, brt, J = 56 Hz, CHF,), 7.3-7.7 (IH, vbrpeak, NH, exchang.), 7.41(1 H, d, J = 7.7 Hz, aromatic H), 7.45 (1 H, t, J = 7.7 Hz, aromatic H), 7.74 (1H, d,J= 7. 7 Hz, aromatic H).

Biological Testing Compounds of Formula (1) were tested for various activities as follows: Screening strategy: The screening strategy is designed to select conipounds with appropriate sodiuni channel blocking activity and low side effect liability. To this end all compounds are processed through the primary sodiuni channel assay (veratrine-evoked uptake of [4C]guanidine into rat forebrain synaptosomes) and IC50 values computed from generated concentration-effect curves. In order to complement this data 1C50's for selected compounds to inhibit binding of[H]BTX-B are also measured.

Previous studies have shown that substituted triazines are potential inhibitors of 5.iydro[olate eductase (DHFR) activity (McCullough and Bertino 1971, Cashmore eta!, 1975, Booth eta!, 1987) and Sapse eta!, 1994). Inhibitors of DHFR (such as Methotrexate) have been used for the treatment of various cancers (Suster et a!, 1978 and Niculescu-Duvaz eta!, 1982) as inhibition ofthis enzyme interferes with cell growth but because of this effect (on cell growth) inhibitors of DHFR may also be teratogenic (Skalko and Gold, 1974, Feldcamp and Carey, 1993 and Buckley et a!, 1997). Should compounds be found which are potent inhibitors of DHFR then such conipounds may, themselves, have potential as anti-cancer agents. Several methods are available for measurement of inhibition of DHFR activity and for this study we have examined effects of compounds to inhibit the binding of [3H] niethotrexate (Myers et a!, 1975 and Rothenberg et a!, 1977).

Another common side-effect marker is inhibition of human ether-a-go-go elated gene potassium (hERG) potassium channel (Inward rectifying, lKr) activity which can be fatal due to heart failure brought about by development of long QT syndrome.

A useful preliminary screen to assess potential to affect this channel is assessed by measurement of inhibition of the binding of {3H]astemizolc to cell membranes expressing hERG. Selected compounds are tested for this activity by measurement of inhibition 10 pM. Assuming inhibition values lie between 10% and 90% it is possible to compute an extrapolated IC50 for each compound.

The above screening cascade identifies compounds with appropriate sodium channel blocking activities that have a low(er) propensity for aforementioned side-effect liabilities. In order to develop these compounds further, sonic knowledge of their pharmacodynamic properties pharmacological action is required.

Sodium channel blockers, such as Sipatrigine, which both reduces the neurological deficit and infarct volume afier middle cerebral artery occlusion in rats (Smith et al, 1997) and phenyloin, (which protect retinal ganglion cell death in an experimental model of glaucoma (Hams and Waxman, 2005) show neuroprotective efficacy in a range of models of nerve degeneration.. As failure of oxygen supply compromises both glycolysis and oxidative phosphorylation, ischaemic daniage ultimately leads to electrical failure (nerve signalling) and pump failure (restoration of cellular membrane potentials). These failures (of electrical and ion pump activity) are associated with decreased local concentrations of ATP (Astrup et al 1981). Thus the effect of compounds to niaintain concentrations of ATP in 0.4 mm slices of rat hippocampus following a severe metabolic insult (incubation with the metabolic inhibitor, lodoacetate) was developed.

Experimental procedures: Preparation of rat forebrain synaptosomes and homogenates: Experiments were performed using forebrain (whole brain less cerebellum/medulla) froni Male Wistar rats weighing 175-250g. All efforts were made to reduce the number of animals used and all experiments were carried out in accordance with the UK Animals (Scientific Procedures) Act, 1986 and the European Community Council Directive of 24 November 1986 (86/609/EEC). Following killing of animals by stunning and decapitation, the forebrain (whole brain less cerebellum/medulla) was rapidly dissected and transferred to a weighed tube containing ice-cold 0.25M sucrose.

Synaptosornes (heavy and light mitochondrial fraction containing synaptosomes) were prepared by transferring the forebrain (of known wet weight) to a glass Potter vessel to which 9 volumes ice-cold 0.25M sucrose had been added and homogenising, using a teflon pestle, by 8 up and down strokes' of a Braun Potter S motor driven homogeniser set to 900rpm. The resulting homogenate was centrifuged at 1036 x g at 4 for 10 mm and the supematant collected. The remaining pellet was resuspended, as above, in fresh ice-cold 0.25M sucrose and the centrifugation step repeated. The supernatant fractions were pooled and centrifuged at 40,000 x g (average) at 4 for 15 mm and the resulting pellet resuspended in the appropriate assay buffer at a concentration of 20-25 mg wet weight per ml appropriate assay buffer.

Homogenates were prepared by transferring the known weight of forebrain to a cooled tube containing 9 volumes of ice-cold 50mM pH 7.4 HEPES buffer. The mixture was homogenised, 40 by 3 x 5 sec bursts of an Ultra-TurraxTM homogeniser set at maximum speed. The resulting homogenate was centrifuged at 40,000 x g (average) at 40 for I 5 mm and the supernatant discarded. The resulting pellet was resuspended in 9 volumes of fresh ice-cold pH 7.4 buffer (as above), the centrifugation step was repeated and the resulting pellet resuspended in the [3H]BTX-B binding buffer at a concentration of 20-25 mg wet weight per ml assay buffer.

I'4C1 guanidine flux and binding of I3HIBTX-B: Both assays were carried out using l4rnl polypropylene test tubes to which a range of concentrations of the compounds under test were added. Test compounds were dissolved in DMSO and added to assays such that maximum concentration of DMSO did not exceed 2% v/v.

[4Clguanidine flux: Compounds under test were pre-incubated for 10 mm at 30 in incubation buffer (50mM pH 7.4 HEPES (adjusted to pH 7.4 with Tris base), 130mM choline chloride, 5.5mM D-glucose, 0.8rnmMgSO4 and 5mM KCI) containing 7.5mg original wet weight of tissue and lOOjtg veratrine HCI in a final volume of 0.Snil. Uptake was initiated by the addition of 0.5m1 of [4C]guanidine (1.OjtCi/ml in incubation buffer) and temiinated 2.5 mm later by the addition of lOml of ice-cold wash buffer (163mM choline chloride, 1.8mM CaC!2 and 0.8mM MgSO4 in 5mM pH 7.4 HEPES buffer), followed immediately by vacuum filtration through Whatman GF/C glass fibre filters using a BrandelTM cell harvester. A further 2 x 5nil of ice-cold wash builer was added to each tube and the vacuum filtration step repeated. The GF/C glass fibre filters were transferred to niinivials and 4nil Pico!luor4 liquid scintillant added using a BrandelTM deposit/dispense system. Radioactivity was measured using a Beckman Liquid Scintillation Counter.

Binding of I3HIBTX-B: Binding was initiated by the addition of 5 rng original wet weight of tissue to tubes containing [3H] BTX-B (incubation concentration measured independently by measurement of radioactivity), drug under test and 25pg a-scorpion venoni in a final volume of 0.25ml of incubation buffer (see above but modified to contain 134mM choline chloride and 1mM KCI). Samples were mixed, incubated for 90 minutes @ 25 and assays terminated by the addition of 5ml of ice-cold wash butler (see above), followed immediately by vacuum filtration through Whatman GF/C glass fibre filters using a BrandelTM cell harvester. A further Srnl of ice-cold wash buffer was added to each tube and the vacuum filtration step repeated. The GF/C glass fibre filters were transferred to minivials and 4m1 Picofluor4 liquid scintillant added using a BrandelTM deposit/dispense system. Radioactivity was nieasured using a Beckman Liquid Scintillation Counter and cprn converted directly to dpm via reference to appropriate quench parameters..

IS

Binding of I3lllMethotrexate All steps were carried out at 4 (or on ice). Freshly dissected rat liver was dissected into 0.25M ice- cold Sucrose and subsequently hornogenised (U-turrax) in 50 mM pH 6.0 phosphate buffer (10 mug tissue) containing 15 mM Dithiothreitol. The resulting homogenate was centrifuged @ 47,500 x g for 20 mm and supernatant (filtered through cotton wool to remove fatty lumps) stored -80 before use (Rothenberg et al).

Inhibition of the binding of [HJmcthotrexate to rat liver homogenate supernatant fractions were carried out essentially as described by Arons et al, 1975. Briefly compounds were incubated with NADPH (480 pM), liver supernatant (DHFR enLyme) and [H]rnetotrexate (50 nM) in a final volume of 410 pL of 50mM pH 6.0 phosphate butler in the presence of Mercaptoethanol (60 mM) for 15 minutes at rooni temperature. The binding reaction was stopped by the addition of 50 pl of a charcoal suspension (consisting of charcoal, Bovine Serum Albumin and Dextran present in a weight ratio of 100:4: 1 suspended in 50 niM pH 6.0 phosphate buffer. Samples were vortexed, allowed to stand for 2 minutes and charcoal was precipated by microcentrifugation full speed for 5 mm. Aliquots of clear supematant were transferred to counting vials containing liquid scintillant for measurement of radioactivity using Liquid Scintillation Spectroscopy.

Specific binding of [3H]rnethotrexate was determined as the difference in binding in the presence and absence of 200 tM cold' niethotrexate. Percentage inhibition values were calculated by comparison to this value.

Computation of IC50 values: Data are presented as mean + sem of number of experiments indicated in brackets.

IC50 values were obtained from radioligand displacement or guanidine flux inhibition curves by plotting logio concentration vs bound ligand/guanidine uptake according the equation:-y = Rniin + Rsp/ l+exp [n (x-C)fl where y = bound (dpm) x = logio compound concentration Rrnin = lower asymptote (i.e. 100% inhibition) Rsp = upper asymptote -Rmin (i.e. specific binding) n = slope (loge) and C = IC50 (i.e. concentration required to inhibit 50 A of specific binding) Hippocampal slice assay: Following killing of animals by stunning and decapitation, the forebrain (whole brain less cerebellum/medulla) was rapidly dissected and transferred to a vessel containing ice-cold pre-gassed artificial cerebral spinal fluid (aCSF). Hippocampi were rapidly dissected and 0.4 mm slices prepared using a Mcllwain tissue chopper. Slices were randomly distributed in 50 ml conical flasks containing 25-30 ml ice-cold pre-gassed aCSF. Flasks were incubated @ 30 for 30 minutes under continued gassing with 95% 02/5% C0 when n-tedium was removed by vacuum aspiration.. Fresh aCSF was added and slices were incubated for a further 30 minutes as described previously.

Medium was again removed by vacuum aspiration and replaced by 25 ml of pre-warmed (300) Ca2-free aCSF. Following a furthcrl() mm incubation under continuous gassing 2 -3 slices were removed (in a volume of 100 ti using an Eppendorf pipette) for measurement of ATP and protein by immediate transfer to individual microfuge tubes containing 0.4 nil of ice-cold 0.5M TriChioroacetic Acid (TCA) lodoacetate (25 p1 of an 0.4 M solution) was added to flask and gassing discontinued. Exactly II nun later 3 -4 slices were removed and transferred to micro fuge tubes as described previously.

Measurement of ATP and protein: Individual slices were disrupted by ultra-sonication and the resulting homogenates centrifuged (à 10000 x g for 5 mm 4o* The supernatant was decanted into a fresh tube and any remaining supernatant removed by vacuum aspiration. The pellet was resuspended in 0.5 ml 0.1 M KOH by ultra-sonication and the resulting suspensions warmed with gentle agitation ( 37 for 30 minutes.

Concentrations of ATP were measured in 6 j.tI of supernatant by mixing with Luciferase reagent (ATPLite from Perkin Elmer) and measuring subsequent luminescence in a 96-well plate Counter.

Protein concentration was measured using BCATM protein assay (Pierce) withBovinc Serum albumin as reference standard.

ATP concentrations were expressed as nmolcs/ mg protein and neuroprotective indices (% protection) calculated by direct comparison with the e!Thct of I tM TTX. hERG:

Assays were carried out to measure effects of compounds @ 10 tM. Making the assumption that binding slopes would be I and for compounds with inhibition values between 10% and 9O% IC50 values were extrapolated.

Results:

Table 1:

Compound l'4C1 guanidine uptake IHIBTX-B binding (synaptosomes) (synaptosomes) Lamotrigine 186 5 26 5 (5) 82.6 2.3 (3) N-methyl-Iamotrigine 35.9 1.8 (3) 7.4 0.4 (2) BW202W92 2.0 0.2 (9) 4.5 0.4 (4) Data presented as mean JCc() (tM) scm of the number of experiments indicated in brackets Table 2: Compound nomenclatures and structures: _____________ 2',3'-dichloro II 2 T,3'-dichloro Methyl 3 2',3'-dichloro Ethyl 4 T,3'-dichloro lso-propyl 2',3-dichloro Hydroxy-ethyl 6 2,3-dichloro n-Propyl 7 2',3'-dichloro Iso-butyl 8 2',3'-dichloro n-Butyl 9 2',3'-dichloro Benzyl 2',3'-dichloro Allyl 11 2,3-dichloro Carhoxamido 12 2',3'-dichloro 4?Mebenzyl 13 T,3'-dichloro 2',3' di-F-benzyl 14 2',3'-dichloro 2'-F-benzyl 2',3'-dichloro 3'-Fbcnzyl 16 2',3'-dichloro 4'-F-benzyl 17 2',3'-diehloro 2'-OMc-benzyl 18 2',3'-dichloro 3'-OMe-benzyl 19 2',3'-dichloro 4'-OMe-benzyl 2',3'-dichloro 3'-Cl-benzyl 21 2',3'-dichloro 4'-Cl-benzyl 22 2',3'-dichloro 2'-CI-benzyl 23 2',3'-dichloro 2'-CFrbenzyl 24 2',3'-dicliloro 3'-CF3-benzyl 2',3'-dichloro 4-CF 3-bcnzyl 26 2',3'-dichloro 2'-F,3'-CF;-benzyl 27 2',3'-dichloro 4'-CON l12-benzyl 28 2',3'-dichloro 4'-CI 1201l-benzyl 29 2',3'-dichloro Thienyl 2',3'dichloro Fury! 31 2',3',5'-trichloro II 32 2',3',5'-trichloro Methyl 33 2',3'-difluoro II 34 2',3',5'-trichloro Propyl 2-F', 3'-Cl H 36 2-F', 3'-C! Methyl 37 2',3'-dichloro 2-BocPipcridCi 12 38 2',3'-dichloro 2-PiperidCll2 39 2',3'-dichloro Cl-12C112CF3 3-F, 2'-Cl 1-1 41 2',5'-dichloro II 42 3',5'-dichloro II 43 Des-chloro H 44 2',4'-dichloro H 2',4'-dichloro Methyl 46 Des-chloro Methyl 47 Des-chloro Ethyl 48 2',5'-dichloro Methyl 49 2',5'-dichloro Ethyl 2',3',5'-trichloro Ethyl 51 2'-OCF3 II 52 2',3'-dichloro F-ethyl 53 3',5'-dichloro Methyl 54 3',S'-dichloro Ethyl 2',3',5'-trichloro F-ethyl 56 2',3',5'-trichloro 3,3,3,-triF-propyl 57 2',3'-dichloro 2,2-diE-ethyl

Table 3: [4C]guanidine flux summary

Compound Mean Scm n ______________ (lC5O: tiM) Sipatrigine 13.1 1.3 10 BW202W92 2.0 0.2 9 Lamotrigine 208.9 38.0 7 2 35.9 1.8 3 3 6.4 0.4 2 4 4.0 0.5 2 172.2 46.2 4 6 2.8 0.5 4 7 2.9 0.3 2 8 2.1 0.2 2 9 4.8 0.4 4 6.1 2.6 2 11 >100 -2 12 4.1 0.5 2 13 2.8 0.0 2 14 2.8 0.6 2 3.9 0.3 2 16 5.1 0.6 2 17 4.6 0.3 2 18 3.1 0.1 2 19 5.0 0.6 2 3.5 0.1 2 21 6.5 0.6 2 22 4.6 -1 23 4.4 -24 4.0 -I 6.2 - 26 5.3 - 27 234.4 -1 28 49.0 -1 29 7.3 1.0 2 4.4 0.2 2 31 16.2 3.3 2 32 6.4 0.4 2 33 >300 -2 34 1.3 -489.8 -1 36 128.8 - 37 12.9 -1 38 20.9 -1 39 3.9 0.3 2 834.5 142.7 2 41 524.9 -1 42 154.9 -1 43 >500 - 44 603.0 -87.1 -I 46 2291.0 - 47 676.0 -1 48 28.2 -49 20.9 -

Table 4: [3H]BTX-B

binding summary ____________________________

Example Assay Tissue prep'n Mean Scm 11 (ICSO: j.iM) Sipatrigine [11]BTX-B Synaptosomes 4.9 0.7 4 [311]BTX-B Homogenate 9.4 2.8 3 BW202W92 {111]BTX-B Synaptosomes 4.5 0.4 4 [II]BTX-B homogenate 4.1 0.6 5 Lamotrigine [II]BTX-B Synaptosomes 76.6 6.2 5 [l1]BTX-B homogenate 97.9 37.7 3 2 [II]BTX-B Synaptosomes 7.2 0.8 4 [3H]BTX-B homogenate 2.5 1.3 3 3 [3H]BTX-B Synaptosomes 3.7 0.6 2 [hh]BTX-B Homogenate 0.9 - 4 [h1]BTX-B Synaptosomes 1.5 0.2 2 [F1]BTX-B Homogenate 0.6 - [3H]BTX-b Synaptosomes25.7 -1 [h1]BTX-b Ihomogenate 7.2 - 6 [hI]BTX-B Synaptosomes 3.6 1.3 2 [3H]BTX-B Homogenate 0.5 - 7 [31l]BTX-B Synaptosomes 1.0 1 8 [11]BTX-B Synaptosomes 1.7 -I 9 [3H]BTX-B Synaptosomes 2.2 -I 29 [Ih]BTX-B Synaptosomes 2.2 -1 [3H]BTX-B Synaptosomes 2.6 - 31 [31-I]BTX-B Synaptosomes 21.9 - 32 [h1]Bi'X-B Synaptosomes 1.9 - 34 [I1]BTX-B Synaptosomes 0.5 -1 39 [hh]BTX-B Synaptosomes 1.6 -1 [3H]B'h'X-B Synaptosomes 282.8 -

Table 5:

Extrapolated lC501s (hERG) from single point inhibition study and [K] (hERG): [Na] channel selectivities \IDS Results Example hERG hERG]tuie [lI]BTX-B JQ hE!i %inhthion Extrapolated I(.,, (riM) IC (tiM) [4C]guaii dine [l BIX-B Sipatrigine 87 I 5 13.1 49 0.1 0.3 I 900 208 9 76 6 > 4.3 > 11 7 2 12 73 3 35 9 7 2 2.0 10.2 3 32 21.2 64 3 7 3.3 5.7 3 36 17.8 40 1.5 4.5 11.9 4 35 18.6 2 8 3.6 6.6 5.2 4 34 194 29 LO 6.7 194 58 7.2 2.1 1.7 3.5 43 6 78 28 4.8 2.2 06 13 14 80 2 5 2.8 nd 0 9 - IS 83 20 3.9 nd 05 - 13 84 19 28 nd 07 - 17 84 1.9 46 nd 04 - 18 87 1.5 31 nd 05 - 23 90 1.1 4.4 nd 03 - 24 94 0.6 4 0 nd 0 2 - 50 100 6 1 nd 16 - 29 83 2.0 7 3 2.2 0.3 0 9 65 5.4 4.4 2.6 1 2 2 1 31 13 669 162 219 41 31 32 33 203 64 1 9 3 2 10.7 34 62 6 1 1 3 0.5 47 12 3 36 33 20.3 128.8 nd 02 - 38 80 2 5 20 9 nd 0 I - 39 40 15.0 39 1.6 38 94 * Synaptosome data

Table 6: Summary of {H]methotrexate binding data

Examples Assay concentration 1Cso Inhibition fl (j.iM) (1.tM) (% assay conc'n) Methotrexate -1 8 0.08 nM -4 Pyrimethamine -2 8 - Trimethopnm -955 -BW202W92 10 --I 99 29 Sipatrigine 10 -0 198 --6 1 (Lamotrigine) -770 96 -3 --1.5 0.5 2 99. 11 1 2 10 -0 99 --3 I 3 10 -0 99 -1 1 4 10 --1 1 99 -0 1 10 --1 1 101 -1 I 6 10 -0 I 99 -2 1 7 JO --4 1 99 --2 1 8 10 --4 99 --2 1 9 10 -2 1 99 --4 1 10 --2 1 99 --I 11 20 --3 1 99 --3 I 12 20 --2 99 --3 1 13 10 --3 I 99 --4 1 14 10 --1 99 --4 1 10 -- 4 1 99 --5 I 16 10 --3 99 --3 I 17 10 --4 1 198 --3 1 18 10 --4 I 99 --5 I 19 10 --2 I 99 --3 I 9 --4 I 99 --7 1 21 10 --4 1 --4 1 29 ->250 - ->250 - 31 -41.0 4.7 -2 32 ->250 -2 33 -441.6 5.1 -2 34 ->30() - -81.3 - 36 ->300 - 37 ->300 - 38 ->300 -1 39 >300 -1 589 - 41 50 -93 1 42 50 -99 1 43 50 -82 1 44 -105 -495 --I 46 288 -1 47 495 -I 1 48 -251 -1 49 -95 - % inhibition computed from methotrexate displacement curve

Table 7: Summary of hippocampal slice data

Example EC() Max Protection EC60 (tM) (% v TTX) (max set ( 100) Sipatrigme 107 90.0 12.6 I (Lamotrigine) 30.2 79.0 33.1 2 36.3 98 5 37. 4 3 16.0 87.() 20.0 6 10.4 81 27 14.5 7 1.3* 47 3 - 8 10.0 46.0 - 9 >>30 -- 31 1.8 91 4 2.2 32 5.9 65 7 - 34 9.8 37.0 -slope set to 2. 30 IATPI in presence of 1 jiM TTX was if max approaches I 00% 71.8 2.9% (17) as then data re-computed compared with untreated control (prc-IOAA slices) = 100% protection. setting max @ 100% Table 8 Effect of compounds on uptake of [1 4C]guanidine Compound Rmin % Specific Hill plC5O 1C50 (jtM) Lamotriginc 1968 597 72.9 0.93 3.75 0.13 177.8 1934 990 72.0 0.71 3.68 0.27 208.9 BW202W92 3019 70 55.9 1.07 5.61 0.04 2.5 2616 91 66.8 0.93 5.52 0.03 3.0 51 2026 27 72.2 1.22 2.76 0.11 1737.8 52 2472 197 64.9 0.74 5.00 0.08 10.0 53 2009 575 72.2 0.73 4.34 0.16 45.7 54 2297 316 65.7 0.76 4.73 0.11 18.6 2209 122 67.9 0.79 5.20 0.05 6.31 56 1807 107 72.9 0.83 2.90 0.20 1258.9 References: McCullough, J. L., and Bertino, R. (1971) Biochem Pharmacol 20(3): 561-74.

Cashmore, A. R., Skeel, R.T., Makulu, D.R., Gralla, E.J. and Bertino, J.R. (1975) Cancer Res 35(1): 17-22.

Booth, R.G., Selassie, C.D., Hansch, C. and Santi, D.V. (1987) J Med Chem 30(7): 1218-24.

Sapse, A.M., Waltham, M.C. and Bertino, J.R. (1994) Cancer Invest 12(5): 469-76.

Niculescu-Duvaz,1., Ciustea, G., Stoicescu, D., Muresan, Z. and Dobrc, V. (1982) Neoplasma 29(1): 43-52.

Suster, D.C., Tarnauceanu, E., Botcz, G., Dobre, V. and Niculescu-Duvaz,I. (1978) J Med Cheni 21(11): 1165-7 Skalko, R.G. and Gold, M.P. (1974) Teratology 9(2):159-63.

Felikamp, M. and Carey, J.C. (1993) Teratology 47(6): 533-9.

Buckley, L.M., Bullaboy, C.A., Leichtrnan, L. and Marquez, M. (1997) Arthritis Rheum40(5): 971-3.

Rothenberg, S.P., da Costa, M. and lqbal, M.P. (1977) Cancer Treat Rep 61: 575-84.

Arons, E., Rothenberg, S.P., da Costa, M., Fischer, C. and lqbal, M.P. (1975) Cancer Research 35: 2033-38 6k

Claims

CLAIMS

I. Use of a conipound of formula (1): ::Ii R6) H2N N NH (1) or a salt or solvate thereot in which R1 is hydrogen, C10 alkyl, C21o alkenyl, C1 alkylaryl, C1 alkyl-heterocyclyl or C..10 cycloalkyl, any of which is optionally substituted by halogen, halo C1, alkyl, C1 alkyl or C1 alkoxy; R2 to R6 are independently selected from hydrogen, halogen, C1 alkyl, alkenyl, alkynyl or alkoxy (all optionally substituted by one or more of halogen, hydroxy and aryl), anhino, mono-or di-substituted amino, alkenyloxy, acyl, acyloxy, cyano, nitro, aryl and alkylthio groups for the preparation of medicaments for use as sodium channel blockers, or as antifolates, or as antimalarials in the treatment of disorders in mammals.

2. Use according to claim 1, for the treatment of epilepsy, multiple sclerosis, glaucoma and uevitis, cerebral traumas and cerebral ischaernias, stroke, head injury, spinal cord injury, surgical trauma, neurodegenerative disorders, motorneurone disease, Alzheirners disease, Parkinsons disease, chronic inflammatory pain, neuropathic pain, migraine, bipolar disorder, mood, anxiety and cognitive disorders, schizophrenia and trigeminal autonomic cephalalgias.

3. Use according to claim 1, for treatment of mammalian cancers.

4. Use according to claim 1, for treatment of malaria.

5. Use according to any one of claims I to 4, in which R1 is Ciio alkyl, C21o alkenyl, C1 alkyl-aryl, or C1 alkyl-heterocyclyl, any of which is optionally substituted by halogen, halo alkyl, halo C1 alkoxy, C1, alkyl or Ci 6alkoxy; 6. Use according to any one of claims I to 5, in which R2 is selected from halogen, C1 alkyl, alkenyl, alkynyl or alkoxy (all optionally substituted by one or more of halogen, hydroxy and aryl), amino, mono-or di-substituted amino, alkenyloxy, acyl, acyloxy, cyano, nitro, aryl and alkylthio groups; and R to R are independently selected from hydrogen, halogen, C1 alkyl, alkenyl, alkynyl or alkoxy (all optionally substituted by one or more of halogen, hydroxy and aryl), amino, mono-or di-substituted amino, alkenyloxy, acyl, acyloxy, cyano, nitro, aryl and alkylthio groups.

7. Usc according to any one of claims I to 4, in which R is hydrogen and R2 to R6 are independently selected from hydrogen, halogen, haloalkyl and haloalkoxy.

8. Use according to any one of clainis ito 4, in which R' is alkyl, hydroxyalkyl, haloalkyl, heterocyclylalkyl, alkenyl, carboxamido, benzyl, bcnzyl substituted by halogen, alkyl, alkoxy, hydroxyalkyl, haloalkyl or carboxaniido and R2 to R' are independently selected froni hydrogen and halogen.

9. Use according to any one of claims I to 4, in which R2 to R' are hydrogen and R' is hydrogen or alkyl.

10. Use according to any one of claims I to 4, in which the compound of formula (I) is selected from 5-amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2-isopropyl-1,2, 4-triazine; 5-amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2-n-propyl-l,2, 4-triazine; 5-amino-6-(2-pentyloxphenyl)-2,3-dihydro-3-imino-2-methyl-I,2,4-triazine; 5-amino-6-(2,3,5-trichlorophenyl)-2,3-dihydro-3-iniino-2-methyl-I,2, 4-triazine; 5-amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2-methyl-I,2, 4-triaLine; 5-amino-6-(2,3-dichlorophenyl)-2,3dihydro-3-imino-2-benzyl-1,2,4-triazine; 5-amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-iniino-2-ethyl-1,2, 4-triazine; 5-amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2-isopropyl-1,2, 4-triazine; 5-arnino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2-n-propyl-i,2, 4-triazine; 5-amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2-isobutyl-I,2, 4-triazine; 5-amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-inhino-2-n-butyl-1,2, 4-triazine; 5-amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2-aIlyI-I,2,4-lriazine; 5-amino-6-(2,3,5-trichlorophcnyl)-2,3-dihydro-3-iniino-2-methyl-I,2, 4-triazine; 5-amino-6-(2,3,5-trichlorophenyl)-2,3-dihydro-3-imino-2-propyl-I,2, 4-triazinc; 5-amino-6-(2-Iluoro,3-chlorophcnyl)-2,3-dihydro-3-iniino-2-methyl-I,2, 4-triazine; 5-aniino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-irnino-2-(3,3, 3-trifluoropropyl)-1,2,4-triazine 5(3)-aniino-6-(2,4-dichlorophenyl)-2,3(2,5)-dihydro-3(5) -irnino-2-mcthyl-1,2,4-triazine; 5(3)-arnino-6-phenyl-2,3(2,5)-dihydro-3(5)-iniino-2-mcthyl-1,2,4-triazine; 5(3)-amino-6-phcnyl-2,3(2,5)-dihydro-3(5)-imino-2-ethyl-I.2,4-triazine; 5(3)-amino-6-(2,5-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2-methyl-I, 2,4-triazine; 5(3)-arnino-6-(2,5-dichlorophcnyl)-2,3(2,5)-dihydro-3(5)-imino-2-ethyl-1, 2,4-triazine; 5(3)-amino-6-(2,3,5-trichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2-ethy-I, 2,4-triazine; 5(3)-arnino-6-(2,3-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2(2-fluoroethyl)-I,2,4-triazine; 5(3)-amino-6-(3,5-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-imino-2-methyl-1, 2,4-triazinc; 5(3)-arnino-6-(3,5-dichlorophenyl)-2,3(2,5)-dihydro-3(5)-irnino-2-ethyl-I, 2,4-triazinc; 5(3)-arnino-6-(2,3,5-trichlorophenyl)-2,3(2,5)-dihydro-3(5)-irnino-2(2-fluorocthyl)-I,2,4-triazine; 5(3)-arnino-6-(2,3,5-trichlorophcnyl)-2,3(2,5)-dihydro-3(5)-imino-2-(3,3, 3-trilluoropropyl)-I,2,4-iriazine; 5(3)-arnino-6-(2,3,dichlorophcny! )-2,3(2,5)-dihydro-3(5)-irnino-2-(2,2-di fluoroethyl)- 1,2,4 triazine.

11. A compound of formula (1): NR1 H2N N NH (1) or a salt or solvate thereof in which R' is C1 alkylaryl or C1 alkyl-heterocyclyl, which is optionally substituted by halogen, halo C1 alkyl, C1 alkyl or C1 alkoxy (excluding unsubstituted benzyl); R2 is selected from halogen, C1 alkyl, alkenyl, alkynyl or alkoxy (all optionally substituted by one or more of halogen, hydroxy and aryl), amino, mono-or di-substituted amino, alkenyloxy, acyl, acyloxy, cyano, nitro, aryl and alkylthio groups; and R3 to R6 are independently selected from hydrogen, halogen, C1 alkyl, alkenyl, alkynyl or alkoxy (all optionally substituted by one or more of halogen, hydroxy and aryl), amino, mono-or di-substituted amino, alkenyloxy, acyl, acyloxy, cyano, nitro, aryl and alkylthio groups.

12. A compound according to claim 11 in which R' is substituted benzyl, piperidinyl-niethyl, luryl-methyl, or thienyl-methyl.

13. A compound according to claim 11, selected from 5-amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2-(2 -fluorophenyl-methyl)- 1,2,4-triazine; 5-amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2-(3 -fluorophenyl-methyl)-I,2,4-triazine; 5-aniino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2(4'-fluorophenyl-methyl)-I,2,4-triazine; 5-amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2-(2,3 -fluorophenyl-methyl)-I,2,4-triazine; 5-amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2-(3 -chlorophenyl-methyl)- 1,2,4-triazine; 5( 5-amino-6-(2,3-dichlorophenyl)-2,3 -dihydro-3-imino-2-(4 -chiorophenyl-methyl)-I,2,4-triazine; 5-arnino-6-(2,3-dichlorophcny! )-2,3-dihydro-3-irnino-2-(4 -methyiphenyl-niethyl)-I,2,4-triazine; 5-amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-irnino-2-(2 -methoxyphcny!-methyl)-I,2,4-triazine 5-arnino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-irnino-2-(3 -methoxyphenyl-meihyl)-I,2,4-triazine; 5-amino-6-(2,3-dichlorophcnyl)-2,3-dihydro-3-imino-2-(4 -methoxyphenyl-methyl)- 1,2,4-triazine; 5-amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2-(2 -chlorophenyl-methyl)-I,2,4-triazine; 5-arnino-6-(2,3-di chioropheny! )-2,3-dihydro-3-imino-2-(2 -trifluoromethyiphenyl-methyl)-! ,2,4-triazine; 5-amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3 -imino-2-(3 -tn fluoromethyiphenyl-methyl)-! ,2,4-triazine; 5-arnino-6-(2,3dichloropheny! )-2,3-dihydro-3-imino-2-(4 -trifluoromethylphenyl-methyl)-I,2,4-triazine; 5-arnino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2-(2 -fluoro-3 -trifluoromethylphenyl-methyl)-1,2,4-lriazine; 5-amino-6-(2,3-dichloropheny!)-2,3-dihydro-3-imino-2-(3-thieny!-nicthyl) -1,2,4-triazinc; 5-aniino-6-(2,3-dichloropheny!)-2,3-dihydro-3-irnino-2-(3-furyl-methy!)-1, 2,4-triazine; 5-amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2(N-boc-piperidin-4-yl-methyl)-I,2,4-triazine; 5-amino-6-(2,3-dichlorophenyl)-2,3-dihydro-3-imino-2(piperidin-4y1-methy!)-1,2,4-triazinc.