AP164A - Pharmacologically active cns compounds - Google Patents

Abstract

A class of substituted phenylpyrimidine compounds are disclosed which are potent inhibitors of the excitatory amino acid, glutamate. Such compounds are useful in the treatment or prevention of a range of cns disorders including celebral ischaemic damage and epilepsy.

Description

PHARMACOLOGICALLY ACTIVE CMS COMPOUNDS

The present invention relates to a class of pyrimidine compounds which are useful in the treatment of central nervous system (CNS) diseases and disorders such as the prevention of cerebral ischaemic damage, to pharmaceutical compositions containing them, to their use in the treatment of such disorders, and to methods of preparing them.

Glutamate is an excitatory amino acid which functions as a neurotransmitter. However, when its extracellular concentration is sufficiently high, glutamate acts as a powerful neurotoxin, capable of killing neurones in the central nervous system, (Rothman & Olney (1986) Prog.Brain.Res., 63, 69). The neurotoxic effect of glutamate has been implicated in a number of central nervous system disorders and disease states including cerebral ischaemic damage, epilepsy and chronic neurodegenerative disorders, such as Alzheimer's* disease, motor system disorders, and Huntington's chorea, (Meldrum Clinical Science (1985) 68 113-122). In addition, glutamate has been implicated in other neurological disorders such as manic depression, depression, schizophrenia, high pressure neurological syndrome, chronic pain, trigeminal neuralgia and migraine.

In European Patent application No.21121 there is disclosed a group of

3.5- diamino-6-(substituted phenyl )-1,2,4-triazines which are active in the treatment of CNS disorders, for example in the treatment of epilepsy. One compound described in that application,

3.5- diami no-6-(2,3-dichlorophenyl )-1,2,4-tri azine (lamotrigine), has been shown to inhibit the release of the excitatory amino acids, glutamate and aspartate, (Leach et £[ Epilepsia 27, 490-497 1986, A.A.Miller et al New anticonvulsant drugs. Ed. Meldrum and Porter 165-177, 1987).

The present inventors have now found that a series of substituted pyrimidine compounds, as defined in Formula I, are potent inhibitors of glutamate release; these comoounds are useful in the treatment of

I!) Ill I) U dV

MJWD/MS/PA1024/22nd November 1989

- 2 the above mentioned disorders and disease states of the central nervous system. The pyrimidine compounds of formula I are also inhibitors of aspartate release.

Thus in a first aspect of the present invention there is provided the use of a compound of Formula I or an acid addition salt thereof in the manufacture of a medicament for treating or preventing CNS disorders or diseases of a mammal, wherein

in Formula I,

R. and R_? are the same or different and are selected from hydrogen, ¹ 4 1111 halo, hydroxy, alkoxy, alkyl, alkylthio and a group NR R where R and rH are the same or different and are selected from hydrogen, alkyl, aryl and arylalkyl or together with the nitrogen atom to which they are attached form a cyclic ring optionally substituted with one or more alkyl groups and optionally containing a further heteroatom;

Rg is hydrogen, alkyl optionally substituted by one or more halo radicals, or is amino, alkylamino, dialkylamino, cyano, nitro, halo, carbamoyl, hydroxy, carboxy, alkoxy, alkylthio, alkylthioalkyl, S(O)_nalkyl, di(alkyloxy)alkyl, -C(R):NOH or -COR or -CO2R wherein R is hydrogen or alkyl, or a group CH^X where X is hydroxy, alkoxy, aryloxy, ary la Iky loxy, halo, cyano, -NrW^ wherein R^ and R^ are as defined above, S(0)_n~alkyl where n is 1 or 2, or SO^NR^R^; each of R^ to Rg are the same or different and each is selected from hydrogen, halo, alkyl, perhaloalky 1, cyano, carbamoyl, carboxy, COR,

MJWD/MS/PA1024/22nd November 1989

- 3 nitro, amino, alkylsulphonylamino, alkoxy, S(0)_n-alkyl where n is 1 or 2, or SO₂NR¹R¹¹; or and R$ or Rg and Rg together are the group -CH=CH-CH=CH- or the group -Cl^-Cl^-C^-Cl·^- in which case both R? and Rg are hydrogen; and optionally one of the nitrogen atoms in the pyrimidine ring may be N alkylated or optionally may be an N oxide;

the foregoing alkyl groups or moieties of alkyl-containing groups having from 1 to 6 carbon atoms, and the aryl groups or aryl moieties of aryl-containing groups having 6 or 10 carbon atoms.

Certain compounds of Formula I are chiral, and it will be appreciated that in these instances, Formula I encompasses both the racemic mixture and the individual enantiomers of such compounds. It will also be appreciated that when one or more of Rp R₂ and Rg are hydroxy, they may also exist in their tautomeric form.

A preferred class of compounds of Formula I which are potent inhibitors of glutamate are those wherein one of and R2 is amino and the other is selected from amino, hydroxy, halo, morpholino, piperazinyl, N-alkylpiperazinyl, Ν,Ν-dialkylamino, N-alkylamino or alkylthio;

Rg is alkyl optionally substituted by one or more halo radicals, or is alkyl, alkylthio, hydrogen, hydroxy, alkoxy, halo, carboxy, carbamoyl, or a group CF^X where X is hydroxy, phenoxy, benzyloxy, alkoxy or alkylthio;

one of and R$ is halo and the other is selected from halo or hydrogen;

R, is halo, hydrogen, nitro or amino;

θ 111 Ry is hydrogen, halo, cyano, alkylthio, SC^NR R alkyl, nitro, amino or methanesuIphonamido; and

Rg is hydrogen or halo.

In the present invention,

R^ is preferably hydroxy, amino, N-alkylamino, Ν,Ν-dialkylamino, morpholino, oioerazinyl, N-alkylpiperaziny1;

MJWD/MS/PA1024/22nd November 1989

- 4 $2 is preferably chloro, amino, N,N-dialkylamino or piperidino;

Rg is preferably hydrogen, alkyl, methoxymethyl, trifluoromethyl, benzyloxymethyl, phenoxymethy1 or methylthiomethyl;

R^ to Rg are preferably selected from hydrogen and chloro. Preferably the alkyl moieties contain from 1 to 4 carbon atoms.

In Formula I, advantageously at least one of R^ and R₂ is amino and the other is amino, piperazinyl or N-methylpiperazinyl, N-alkylamino, N.,N[-dia 1 kylamino; and

Rg is hydrogen, methyl, trifluoromethyl or methoxymethyl.

It is a preferred feature of Formula I that two of R^, Rg and R-, are chloro, in particular it is preferred that all of R^, Rg and R? are chloro. Such compounds are highly potent inhibitors of glutamate release. Preferred examples of the group NR^R^¹ are amino, N-methylamino, N-ethylamino, Ν,Ν-dimethylamino, piperazinyl, N-methylpiperazinyl, piperidinyl, and morpholino.

An especially preferred class of compounds within Formula I are those wherein

R^ is selected from amino, piperazinyl, Ν,-methylpiperazinyl, N-morpholino, Ν,Ν-dimethylamino, and N-ethylamino;

R^ is amino;

Rg is selected from trifluoromethyl, hydrogen, methyl, benzyloxymethyl, methoxymethyl and methylthiomethyl;

R₄ is chloro; and at least one of Rg, Rg and R? is chloro, and the remainder are selected from hydrogen, chloro and nitro; and Rg is hydrogen or R^ and Rg are both hydrogen, Rg and R? are both chloro and Rg is selected from hydrogen chloro and nitro.

The present invention also provides a subclass of compounds of Formula I, which whilst being potent inhibitors of glutamate release show only weak (ie. having an ΙΟ_ςθ of >20μΐη) or insignificant inhibitory effects on the enzyme dihydrcfolate reductase . Accordingly, in a preferred bad

MJWD/MS/PA1024/22nd November 1989

- 5 embodiment of the present invention there is provided compounds of Formula I where R^ to Rg are hereinbefore defined with the proviso that when

Ry is halo, then

Rg is hydrogen, perhaloalkyl, methyl or methoxymethyl and/or Rg is nitro and/or

R^ is N-alkylpiperazinyl, morpholino, Ν,Ν-dimethylamino, piperazinyl or N-ethylamino; or with the proviso that when Rg is chloro then

R^ is halo, and Rg is hydrogen, perhaloalkyl, methoxymethyl, methyl or halo and/or Rj is N-methylpiperazinyl, piperazinyl, morpholino or Ν,Νdimethylamino, or N-ethylamino.

Compounds of Formula I may be used in the treatment or prophylaxis of acute and chronic disorders of the mammalian central nervous system. The acute condition comprises cerebral ischaemia which may arise from a variety of causes including stroke, cardiac arrest, bypass surgery, neonatal anoxia and hypoolycaemia; and also physical injury or trauma of the spinal cord or brain. Chronic neuro- degenerative disorders which may be treated include Alzeheimer's disease, Huntington's chorea, 01ivopontocerebrellar atrophy, motor system disorders. Other neurological conditions which may be treated with a compound of Formula I include depression, manic depression, schizophrenia, chronic pain, epilepsy, trigeminal neuralgia and migraine.

APO00164

In a further aspect, the present invention provides treatment or prevention of a CNS disorder or disease including man, comprising the administration to the non-toxic effective amount of a compound of Formula addition salt thereof.

a method of of a mammal, mammal of a I or an acid

In particular, the present invention provides a method of treating a mammal predisposed to or having neurotoxic extraeellular glutamate

MJWD/MS/PA1024/22nd November 1989

- 6 levels of the central nervous system comprising the administration to the mammal of a non-toxic effective amount of a compound of Formula 1 or an acid addition salt thereof.

Certain substituted phenylpyrimidines of the present invention are known in the art as having antimalarial activity. See for example Brit.J.Pharmacol. 6, 185-200 (1951); JACS, 73, 3763-70, (1951). Other phenylpyrimidines are known from Chem.Biol. Pteridines, 463-468, (1982) and Pharmacotherap.Budesinsky, p.129-141, (1963), ed. Oldrich Hanc.

Nonetheless, certain compounds of the present invention are novel and accordingly the present invention provides a compound of Formula I or an acid addition salt thereof wherein to Rg are as hereindefined, with the proviso that at least one of R^ to Rg is other than hydrogen, and further with the proviso that when R^ and R? are both amino, or when R^ is hydroxy and R? is amino, and Rg is alkyl or hydrogen, and R? is hydrogen, then R^ and Rg are both halo.

Other novel compounds of the present invention are those where R^ to Rg and Rg are as hereinbefore defined and one of or Rg is other than hydrogen, and R? is halo, alkyl, perhaloalkyl, cyano, nitro, amino, alkylthio, S(0)_n-alkyl or SO^NR^R^.

A third class of novel compounds of the present invention are those of Formula I where Ry is morpholino, piperazinyl, N-alkylpiperazinyl, N_, N_-d i a Iky 1 ami no, N-alkylamino or alkylthio and R? to Rg are as hereinbefore defined.

A fourth class of novel compounds of the present invention are those of Formula I where R^ and R? are as hereinbefore defined; and Rg is alkoxy, alkylthio or alkyl substituted by one or more halo radicals, or is a group Ci^X where X is alkylthio, aryloxy, arylalkyloxy, alkyloxy or hydroxy;

MJWD/MS/PA1024/22nd November 1989

- 7 to Rg are the same or different and are each selected from hydrogen, halo, perhaloalkyl, cyano, nitro, amino or alkylthio;

Ry is halo, alkyl, perhaloalkyl, cyano, nitro, amino, or a group SC^NiR¹¹¹^ wherein R¹¹¹ is alkyl; and

Rg is hydrogen or halo.

A fifth class of novel compounds of the present invention are those of Formula I where Rj and R₂ and R^ to Rg are as hereindefined, and R^ is alkoxy, aryloxy, arylalkyloxy or alkylthio.

AP 0 0 0 1 6 4

MJWD/MS/PA1024/22nd November 1989

- 8 Preferred novel compounds of the present invention include the following, the numbers referring to the Examples hereinafter appearing:Example No.

1. 4-Amino-2-(4-methylpiperazin-1-y1)-5-(2,3,5-trichloropheny1)-6trifluoromethylpyrimidine

2. 2,4-Diamino-5-(2,3,5-trichlorophenyl)-6-methoxymethylpyrimidine

3. 4-Amino-2-(4-methylpiperazin-l-yl)-5-(2,3,5-tri chlorophenyl) pyrimidine

4. 2,4-Diami no-5-(2,3,5-tri chlorophenyl )-6-tri fluoromethyl pyrimidine

5. 2,4-Diamino-5-(4-nitro-2,3,5-tri chlorophenyl)pyrimidine

6. 2,4-Diamino-5-(2,3,5-trichlorophenyl)-6-methylpyrimidine

7. 4-Amino-2-N-morpholino-5-(2,3,5-trichlorophenyl )-6-trifluoromethylpyrimidine

8. 4-Ami no-2-N.,N-dimethy1 ami no-5-(2,3,5-tri chi oropheny! )-6trifluoromethylpyrimidine

9. 4-Amino-2-morpholino-N-5-(2,3,5-trichlorophenyl)pyrimidine

10. 4-Amino-2-N,N-dimethylami no-5-(2,3,5-trichloropheny1) pyrimidine

11. 4-Amino-6-methyl-2-(4-methyIpiperazin-l-yl)-5-(2,3,5-tri chlorophenyl) pyrimidine

14. 2,4-Diamino-5-(2,3-dichlorophenyl)-6-trifluoromethylpyrimidine

15. 2,4-Diami no-5-(2,3-dichloropheny1)-6-methy1 pyrimidine

16. 2,4-Diamino-5-(2,3-dichloropheny1)-6-methoxymethylpyrimidine

25. 2,4-Diami no-5-(2,4-dichlorophenyl )-6-trifluoromethyl pyrimidine

26. 6-Benzyloxymethyl-2,4-diami no-5-(2,4-dichloropheny1) pyrimidine

27. 2-N-methy1 piperazinyl-4-amino-5-(2,4-dichioropheny1) pyrimidine

28. 2,4-Diami no-5-(2,5-dichlorophenyl)-6-trifluoromethylpyrimi dine

29. 2,4-Diami no-5-(2,3,5-trichlorophenyl) pyrimidine

36. 4-Amino-5-(3,5-dichlorophenyl)-6-methy1-2-(4-methy1 pi perazin-1 yl)pyrimidine

58. 4-Amino-2-N-ethyl ami no-5-(2,3,5-tri chlorophenyl) pyrimidine

79. 4-Amino-2-N-methylamino-5-(2,3,5-trichloropheny1) pyrimidine

MJWD/MS/PA1024/22nd November 1989

- 9 or an acid addition salt thereof.

Suitable acid addition salts of the compounds of Formula I include those formed with both organic or inorganic acids. Such acid addition salts will normally be pharmaceutically acceptable. Thus, preferred salts include those formed from hydrochloric, hydrobromic, sulphuric, citric, tartaric, phosphoric, lactic, pyruvic, acetic, succinic, oxalic, fumaric, maleic, oxaloacetic, methanesulphonic, ethanesulphonic, jj-toluenesulphonic, benzenesulphonic and isethionic acids. These salts can be made by reacting the compound as the free base with the appropriate acid.

While it is possible for the compounds of Formula I to be administered as the raw chemical, it is preferable to present them as a pharmaceutical formulation. The formulations of the present invention comprise a novel compound of Formula I, as above defined, or a pharmaceutically acceptable salt thereof together with one or more acceptable carriers therefor and optionally other therapeutic ingredients. The carrier(s) must be 'acceptable' in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.

The formulations include those suitable for oral, parenteral (including subcutaneous, intradermal, intramuscular and intravenous), rectal and topical (including dermal, buccal and sublingual) administration although the most suitable route may depend upon for example the condition and disorder of the recipient. The formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. All methods include the step of bringing into association a compound of Formula I or a pharmaceutically acceptable acid addition salt thereof (active ingredient) with the carrier which constitutes one or more accessory ingredients. In general the formulations are prepared by uniformly and intimately bringing into association the active ingredient with

MJWD/MS/PA1024/22nd November 1989

- 10 liquid carriers or finely divided solid carriers or both and then, if necessary, shaping the product into the desired formulation.

Formulations of the present invention suitable for oral administration may be presented as discrete units such as capsules, cachets or tablets each containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or a suspension in an aqueous liquid or a non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion. The active ingredient may also be presented as a bolus, electuary or paste.

A tablet may be made by compression or moulding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active ingredient in a freeflowing form such as a powder or granules, optionally mixed with a binder, lubricant, inert diluent, lubricating, surface active or dispersing agent. Moulded tablets may be made by moulding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. The tablets may optionally be coated or scored and may be formulated so as to provide slow or controlled release of the active ingredient therein.

Formulations for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain antioxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents. The formulations may be presented in unit-dose or multi-dose containers, for example sealed ampoules and vials, and may be stored in a freeze-dried (lyophilised) condition requiring only the addition of the sterile liquid carrier, for example, water-for-injection, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets of the kJnd previously described.

MJWD/MS/PA1024/22nd November 1989

- 11 Formulations for rectal administration may be presented as a suppository with the usual carriers such as cocoa butter or polyethylene glycol.

Formulations for topical administration in the mouth, for example buccallyor sublingually, include lozenges comprising the active ingredient in a flavoured basis such as sucrose and acacia or tragacanth, and pastilles comprising the active ingredient in a basis such as gelatin and glycerin or sucrose and acacia.

Preferred unit dosage formulations are those containing an effective dose, an hereinbelow recited, or an appropriate fraction thereof, of the active ingredient.

It should be understood that in addition to the ingredients particularly mentioned above, the formulations of this invention may include other agents conventional in the art having regard to the type of formulation in question, for example those suitable for oral administration may include flavouring agents.

Tablets or other forms of presentation provided in discrete units may conveniently contain an amount of compound of the Formula I which is effective at such dosage or as a multiple of the same, for instance, units containing 5mg to 500mg, usually around lOmg to 250mg.

The compounds of the Formula I are preferably used to treat CNS disorders or diseases by oral administration or injection (intraparenteral or subcutaneous). The precise amount of compound administered to a patient will be the responsibility of the attendant physician. However the dose employed will depend on a number of factors, including the age and sex of the patient, the precise disorder being treated, and its severity. Thus for example when treating a patient with epilepsy the dose range is likely to be significantly lower than when treating a patient after stroke to

APO 0016 4

MJWD/MS/PA1024/22nd November 1989

- 12 alleviate cerebral ischaemic damage. Also the route of administration is likely to vary depending on the condition and its severity.

The compounds of the Formula I may be administered orally or via injection at a dose of from 0.1 to 30mg/kg per day. The dose range for adult humans is generally from 8 to 2,400 mg/day and preferably 35 to 1,050 mg/day. As certain compounds of the Formula I are long acting, it may be advantageous to administer an initial dose of 70 to 2,400 mg the first day then a lower dose of 20 to 1,200 mg on subsequent days.

Examples of such long acting compounds are:

2.4- diamino-5-(2,3,5-trichlorophenyl)-6-trifluoromethylpyrimidine;

4-amino-2-(4-methylpiperazin-l-yl)-5-(2,3,5-trichlorophenyl )-6tr if1uoromethy1pyrimidine; and

4-amino-2-(4-methylpiperazin-l-yl)-5-(2,3,5-trichlorophenyl )pyr i mi di ne.

Long acting compounds in the clinic are advantageous because they are easier to manage. In the chronic situation, they may be administered without infusion and there is the minimum of direct, medical intervention; also in acute conditions, patient compliance is encouraged by minimising daily dosing. Conversely, short acting compounds such as:

2.4- diamino-5-(2,3,5-trichlorophenyl)-6-methoxymethylpyrimidine permit the clinician to control the pharmacological effect of the compound with great precision, since such compounds will be cleared from the central nervous system rapidly. _

MJWD/MS/PA1024/22nd November 1989

- 13 Compounds of the present invention may be made in any manner known to make analogous compounds known in the art (eg. JACS vol 73 (1951) 3763-70).

The present invention also provides a process for the preparation of a compound of Formula I or an acid addition salt thereof, which comprises the reaction of a compound of Formula II * u

I

I Ϊ hi

I

II

L wherein to βθ are as hereinbefore defined, L is a leaving group, and Y is cyano or carboxy, carbonyl, or alkoxycarbonyl with a compound or salt thereof of formula III

KJH n ni

M 2 — C , wherein R^ is as hereindefined, and isolating the compound of Formula I as the free base or an acid addition salt thereof, and optionally converting the base into an acid addition salt thereof or into another acid addition salt, or into another compound of Formula I or an acid addition salt thereof.

V9 I 00 OdV

It will be appreciated that certain compounds of Formula III for example where Rj is hydroxy exists in the corresponding tautomeric form (eg. as urea).

As examples of interconversion of compounds of Formula I, when in the product of the above process one of Rp R₂ or R^ is hydroxy the compound may be halogenated for example using the Vilsmeier Haack reagent or phosphorus oxychloride (POC1), to the corresponding halo compound. This compound may be further converted into a compound of Formula I R,, % or R-, is alkvlthic o, reaction with the

MJWD/MS/PA1024/22nd November 1989

BAD ORIGINAL

- 14 appropriate a lkylthiolate or to the corresponding amino compound (R, , R? or Rg being NR R ) by reaction with the appropriate amino compound in a suitable solvent such as an alkanol, eg. ethanol, or converted to a compound where R^, R₂ or Rg is alkoxy by reaction with appropriate alkoxide.

If it is required to make a compound of Formula I in which one of R.

to Rg is nitro, this can be made from the corresponding compound of Formula I where one of R^ to Rg is hydrogen by utilising standard nitration conditions, eg. sulphuric acid and potassium nitrate, and then further converted by standard reduction means to the corresponding amino compound, eg. utilising PtO₂, AcOH, H_2>

It will be appreciated that amino or halo compounds can be further converted to R^ to Rg as herewithin defined by standard interconversion, for example, via the diazonium salts. When Rg is alkyl this may be converted into perhaloalkyl, or a halogenated alkyl moiety by reaction with the appropriate halogen or N-halo succinimide

*. (NXS) in a suitable solvent such as acetic acid.

>

Compounds in which Rg is CH(0Et)₂ may be converted into the corresponding aldehyde by hydrolysis with dilute acid(s) and thereafter reduced to the corresponding alcohol with a standard reducing agent (eg. sodium borahydride NaBH^), or converted to the corresponding oxime (hydroxylamine hydrochloride in ethanol), which in turn can be converted to the corresponding cyano compound utilising, for example trifluoroacetic anhydride (TFAA), and then to the corresponding amido compound utilising concentrated sulphuric acid. Alternatively, the aldehyde may be oxidised with KMnO^ (potassium permanganate) to give the corresponding acid, which may in turn be reacted with an alcohol to give the corresponding ester.

Where in the product of the above process Rg is a group CH₂OR where R is alkyl or arylalkyl this product may be converted to CHgX by reaction with HX (X = halo) in, <or example acetic acid, and th^;s

MJWD/MS/PA1024/22nd November 1989 bad original further converted to the corresponding cyano compound, for example by treatment with sodium cyanide and DMF, or to fluoromethyl by treatment with for example cesium fluoride (CsF) or to a group CH^NR^R^ by reaction with the appropriate amine. Alternatively, the group Cl^OR can be dealkylated to give the corresponding alcohol, for example with Me^Sil, and this further converted to fluoromethyl with diethylaminosulphur trifluoride (DAST).

Where Rg to Rg contains an alkylthio moiety, this can be oxidised to the corresponding_ sulphoxide and sulphone using for example MCPBA (metachloroperbenzoic acid).

It will be appreciated that other interconversions may be effected as required by those skilled in the art using standard methodologies.

Examples of suitable leaving groups (L) include C, . alkoxy, halo,

11 ^1-4 NR R as hereindefined eg. anilino, morpholino, alkylamino, benzylamino, or alkylthio. Preferably in Formula III, R. is hydroxy, 111 ¹ alkoxy, alkylthio, or a group NR R as herein defined. Advantageously R^ is amino, alkylamino, dialkylamino, piperazinyl or N-methylpiperazinyl.

AP 0 (1 0 1 6 4

Preferably the reaction of the compound of Formula I and II is carried out in a non-aqueous solvent, for example an alkanol, eg. ethanol at elevated temperatures (eg. between 50 to 110°C) in a base, preferably an alkanoxide, preferably under reflux using sodium ethoxide as the base.

Alternatively a compound of Formula I or an acid addition salt thereof may also be made by the reaction of a compound of Formula III with a compound of Formula IV

IV

MJWD/MS/PA1024/22nd November 1989

wherein Rg to Rg and Y are as herein defined. The reaction preferably takes place in a non-aqueous solvent, eg. alkanols, such as ethanol, and at elevated temperatures, preferably under reflux.

Compounds of formula IV can be made by methods known in the art (JACS, 1951, 73, 3763-3770).

Compounds of Formula II may be made by methods known in the art (JACS supra) for example by the reaction of a compound of Formula IV with diazomethane or with alkylorthoesters (JACS, 1952, 74, 1310-1313), or by condensation with an amine.

In Formula III when R^ is piperazinyl or alkyl piperazinyl these can be made by standard methods for example by reaction of a known compound of Formula III where R^ is alkylthio with the appropriate amine, eg. N-methylpiperazine. This reaction preferably takes place at room temperature in water.

Compounds of formula I may also be made by the reaction of a compound of Formula V c

wherein Y and R^ are as hereinbefore defined and R^ and Rj, are alkyl or collectively form a group (CR₉)_n where n is 2 to 4 and R is ‘4 or

-’MJWD/MS/PA1024/22nd November 1989 alkyl, with a compound of Formula III. Most preferably R^ is amino, piperazinyl or methyl piperazinyl. Preferably the reaction is carried out in a non-aqueous solvent, eg. ethanol, under reflux using sodium ethoxide as the base.

Compounds of Formula I may also be prepared from the corresponding dihydropyrimidine by utilising standard dehydrogenation conditions, (eg. JCS, 1956, 1019).

Such dihydropyrimidjne can be prepared by the reaction of a compound of Formula II where Rg to Rg are as defined and L is hydrogen with a compound of Formula III.

APO 0016 4

MJWD/MS/PA1024/22nd November 1989

- 18 In the Examples of the invention set forth below, the chemical and other abbreviations used are standard in the art and have these meanings:-

Na8H4	sodium borohydride
chci3	chloroform
NaHC03	sodium bicarbonate
MgS04	magnesium sulphate
PBr3	phosphorus tribromide
DMF	dimethyl formamide
KCN	potassium cyanide
Et2O	diethyl ether
NaOEt	sodium ethoxide
EtOH	ethanol
H2so4	sulphuric acid
AcOH	acetic acid
MeOH	methanol
N2	nitrogen
HC1	hydrochloric acid
NaOH	sodium hydroxide
Si02	si 1ica
DMSO	dimethylsulphoxide
Na	sodium
DME	dimethoxyethane
Mel	methyl iodide (iodomethane)
EtOAc	ethyl acetate
ch2ci2	dichloromethane ~
- *- r'	triethylamine

MJWD/MS/PA1024/22nd November 1989

- 19 MeNH₂ nh₄oh soci₂

THF

NaH ^CC14

OHFR pto₂

NXS *2

TFAA

CsF Me₃SiI DAST

MCPBA

AIBN methyl amine ammonium hydroxide thionyl chloride tetrahydrofuran sodium hydride carbon tetrachloride dihydrofolate reductase platinum oxide (Adams’ catalyst)

N-halo succinimide halogen trifluoroacetic anhydride cesium fluoride trimethylsilyliodide diethylaminosulphur trifluoride metachloroperbenzoic acid a,q'-azoisobutyronitr i le (2,2‘-azobis(2-methylpropionitrile)

MJWD/MS/PA1024/22nd November 1989

- 20 Exawl· 1

Preparation of 4-Amino-2-(4-methylpiperaz in-1-yl)-5-(2,3,5-tri chlorophenyl )-6-trifluoromethylpyrimidine

1. Preparation of N-roethylpiperazinoformamidine hydriodide

Thiourea (10.8g) was dissolved in acetone (250ml) at 50°C. Iodomethane (10ml) was added and the reaction was stirred at 50°C for 4 hours. After cooling, the solution was diluted with ether (1 litre) and the methiodide salt was filtered, washed with ether and dried in vacuo. 29.2g, 113-115°C. The methiodide salt (5g) was dissolved in water, (30ml) and N-methylpiperazine was added. The solution was stirred, with nitrogen bubbled through, at room temperature for 24 hours. The solution was concentrated in vacuo. The residue was slurried with ethanol, filtered and dried in vacuo. 4.98g, m.pt.230-242°C.

2. Preparation of 2.3.5-trichlorobenzylaIcohol

To a solution of 2,3,5-trichlorobenzaldehyde (Aldrich, 50gms) in ethanol (1.0L) at room temperature was added NaBH^ (7.00gms) and the resulting mixture stirred for 3.5 hours. The reaction was quenched with water, and the solvent evaporated in vacuo before partitioning the residue between CHCl^ and saturated NaHCO^ solution. The organic phase was washed with brine, dried over MgSO^, filtered and the solvent evaporated in vacuo to leave a white solid. 43.00gms, mp. 90-93°C.

3. Preparation of 2.3,5-trichlorohenzyl bromide

To a solution of the alcohol in benzene (400ml) under was added PBr^ (126.58gms), and the mixture stirred at 55-60°C for 3.5 hours. After cooling, the mixture was poured onto crushed ice (2L) and the benzene layer separated. The aqueous phase was washed with benzene (x3) and the combined benzene extracts washed with saturated NaHCO^ solution and water, dried over MgSO^,

MJWD/MS/PA1024/22nd November 1989

- 21 filtered and the solvent evaporated to leave a brownish liquid which solidified on standing, 37.53gms, mp. 40-42°C.

4. Preparation of 2.3.5-trichlorophenylacetonitri le

The bromide was suspended in DMF (130ml)/ water(86.67ml) at 0°C and KCN(12.99gms) added in portions. After stirring at 30-35° for 3 hours, the suspension was diluted with water and extracted with Et₂O. The combined ether extracts were washed with water, dried over MgSO^, filtered and the solvent evaporated in vacuo. Chromatography on silica gel eluting with hexane to 20% ether-hexane gave the desired product as a white solid, 18.52gms, mp. 60-62°C.

5. Preparation of 2-(2.3.5-trichlorophenyl)-4,4.4-trifluoro-3-oxobutyronitrile

To a solution of NaOEt (from 1.04gms Na) in EtOH (60ml) at room temperature under N₂ was added the nitrile (8.40gms) followed by ethyl trifluoroacetate (6.57gms) and the mixture stirred at reflux for 5 hours. After cooling, the solvent was removed jn vacuo and the residue dissolved in water. The aqueous phase was washed with Et₂O (discarded), acidified with i^SO^ and extracted with Et₂0. The combined Et₂0 extracts were washed with water, dried over MgSO^, filtered and the solvent evaporated in vacuo to leave an oil. This was triturated with petroleum ether, and the solid filtered off and dried'. The solid was azeotroped with toluene (x5), 4.89gms, mp. 160-163°C.

6. Preparation of 2-(2.3.5-trichloroph&nyl)-4.4,4-trifluoro-3methoxybut-2-enonitrile

To a solution of the trifluoromethyl ketone in Et₂O (39.62ml) at room temperature was added diazomethane (from 8.55gms Diazald) in Et₂O (79.62ml), and the resulting mixture left to stand at room temperature overnight. Excess diazomethane was then removed in vacuo into AcOH, and the residue was dissolved jn Et₂O, dried

AP 0 0 0 1 6 4

MJWD/MS/PA1024/22nd November 1989

- 22 over MgSO^, filtered and the solvent evaporated in vacuo to leave a brownish oil, 5.20gms.

7. Preparation of 4-Amino-2-(4-methylpiperazin-l-yl)-5-(2,3,5trichlorophenyl)-6-trifluoromethylpyrimidine To a solution of NaOEt (from 0.144g of Na) in EtOH (12.5ml) was added N-methylpiperazinoformamidine hydriodide (1.39g). After stirring for 10 minutes at room temperature a solution of the above intermediate (0.85g) in EtOH (2.5ml) was added and the resulting mixture was stirred at reflux for 4.5 hours. After cooling, the suspension was filtered, and the filtrate was evaporated to dryness in vacuo. Chromatography on silica gel, eluting with CHCl^ -4% MeOH/CHCl^, gave the desired product which was triturated with petroleum ether (b.p. 40-60°C) and dried in vacuo. 0.56g, m.pt 127-129°C.

8. 4-Amino-2-(4-methyIpiperazin-l-yl)-5-(2,3,5-trichlorophenyl)-6trifluoromethylpyrimidine methanesulphonate

The phenyl pyrimidine base (9.6g) was dissolved in absolute ethanol, cooled at 0°C, and methanesulphonic acid (2.14g, 1.62ml) was added. After stirring at room temperature for 2 hours, the solution was evaporated to dryness and the residue triturated with Et₂0, filtered and dried in vacuo to leave a beige coloured solid. This was dissolved in water (500mls) and freeze-dried to leave 10.7g as a tan coloured solid. The methanesulphonate salt could be further purified by triturating with ^BuOH (30ml), filtering, dissolving in water and again freeze drying to leave the title product as a off-white solid. 8.33g mp. 145-7°C.

MJHD/MS/PA1024/22nd November 1989

- 23 Ex—pi· 2

Preparation of 2,4-0iamino-5-(2,3.5-trichlorophenyl)-6-methoxymethyl pyrimidine

A. 2-(2,3.5-tri chlorophenyl )-4-methoxv-3-oxo-butyronitri Ie

To a stirred refluxing solution of sodium ethoxide (from 1.38g sodium) in ethanol (25ml) was added over 5 minutes a mixture of

2,3,5-trichlorophenylacetonitrile (llg) and ethyl methoxyacetate (8.85g) in dry OME (25ml). After 4 hours the mixture was cooled on ice and acidified by dropwise addition of acetic acid (ca. 6ml), diluted with ice-water (150ml) and extracted with dichloromethane (2 x 100ml). The dichloromethane extract was washed with water, dried over MgSO^ and concentrated giving a yellow solid, which was triturated with a little ether and filtered. 8.6q Homogeneous by TLC (19:1 CH^ClgiMeOH).

B. Preparation of 2.4-0iamino-5-(2.3.5-trich1orophenyl)-6methoxymethylpyrimidine

A suspension of the crude acyl acetonitrile (8.5g) in ether (100ml), cooled on ice was treated with an excess of an alcohol free solution of diazomethane in ether (0.035M). After one hour TLC (19:1 CHzC^-'MeOH) showed no starting material. The solution was concentrated to give .a brown waxy solid, which was used without further purification.

To a solution of sodium ethoxide (from 0.76g sodium) in ethanol (30ml) was added guanidine hydrochloride (2.9g). After 15 minutes a solution of the crude enol ether in ethanol (25ml) was added and the mixture refluxed with stirring for 4 hours, cooled and concentrated. The residue was shaken with 2M NaOH (150ml) and the dark solid filtered off, washed with water, dried in air and recrystallised from ethanol (150ml).

5a M.pt. 214-216°C. TLC (1:9 MeOH:CHCl₃) Rf~O.35.

APO0016 4

MJW0/MS/PA1024/22nd November 1989

- 24 C. 2,4-Diamino-5-(2.3,5-tri chlorophenyl)-6-methoxymethylpyriroidine ethanesulphate

To a stirred suspension of the phenylpyrimidine (2g) in ethanol (75ml) was added dropwise ethanesulphonic acid (0.67g) jn ethanol (10ml). After ca 30 minutes the solution became cloudy. Stirring was continued for a further 1.5 hours and the solvent then concentrated to ca 20mls. Ether was added, the solid filtered off and washed with Et₂O before drying jn vacuo, 2.17g, m.pt. 265-268°C.

Example 3

Synthesis of 4-Amino-2-(4-methylpiperazin-1-y1)-5-(2,3,5-trich lorophenyl) pyrimidine mesylate

1. Preparation of 2-(2,3.5-trichloropheny1)-3-oxo-propionitrile, sodium salt

To a solution of NaOEt (from 0.803g of sodium) in ethanol (55ml) cooled in ice, under nitrogen, was added 2,3,5-trichlorophenyl acetonitrile (see Example 1.4). Ethyl formate (5.1ml) was added and the mixture was stirred at room temperature overnight. After stirring for a further 2.5-hours at 50°C, the mixture was cooled and filtered. The filtrate was evaporated, and the residue was triturated with diethyl ether, filtered and dried (6.82g).

2. Preparation of 2-(2,3.5-trichlorophenv1)-3-methoxy-acrylonitrile

The above solid was dissolved in DMF (36ml) and methyl iodide (2ml) was added. The reaction vessel was sealed before stirring the contents at 40°C for 3 hours. The solvent was then evaporated. The residue was partitioned between water and ethyl acetate. The organic phase was washed with water, dried (MgSO^) and the solvent evaporated to give the crude product as a red-brown oil that solidified on standing (5.04g)^MJWD/MS/PA1024/22nd November 1989

- 25 3. Preparation of 4-Amino-2-(4-methylpiperazin-l-yl)-5-(2.3.5trichlorophenyl) pyrimidine

To a solution of NaOEt (from 0.21g of sodium) in ethanol (20ml) was added N-metbylpiperazinoformamidine hydriodide (2.06g) (see Example 1.1). After stirring for a further 10 minutes, the compound of Example 3.2 (lg) was added and the mixture was The mixture was left standing at then filtered. The filtrate was concentrated and the residue was purified by chromatography on Si0₂, eluting with CHC1to 4% MeOH/CHCl^ to give the title compound as the free base. 0.89g, mp. 162-164°C.

stirred at reflux for 4 hours, room temperature overnight and

The free base (0.805g) was then dissolved in ethanol (35ml) and cooled in an ice bath. Methanesulphonic acid (0.21g) was added and the reaction was stirred at room temperature for 2 hours. The solvent was then evaporated and the residue was triturated with diethyl ether, filtered, dissolved in cold water and freeze dried to give the title salt as a pale green solid, 0.98g, mp. 143-146°C.

Example 4

Synthesis of 2.4-Diaroino-5-(2.3.5-trich1orophenyl)-6-trifluoromethyl pyrimidine

To a solution of NaOEt (from 0.88g of Na) in EtOH (82ml) was added guanidine hydrochloride (2.98g). The resulting white suspension was stirred at room-temperature for 10 minutes. A solution of the enol ether (Example 1.6) in ethanol (27ml) was added and the resulting mixture stirred at reflux for 4.25 hours. After cooling, the suspension was filtered, and the filtrate evaporated to dryness in vacuo. Chromatography on silica gel eluting with CHCl^ to 2% MeOH:CHCl^ gave the desired product which was triturated with Et^O and dried in vacuo, l./Sgms, m.p. 226-227°C. _

MJW0/MS/PA1024/22nd November 1989

- 26 Exampl· 5

Preparation of 2.4-Diamino-5-(4-nitro-2.3.5-trichloropheny1) pyrimidine

The compound from Example 29 was dissolved in concentrated sulphuric acid (2.5ml), potassium nitrate (25.8mg) added and the solution stirred for 3 hours. The solution was then poured onto ice and basified with 10 N NaOH. The product was extracted with ethyl acetate (x3), dried over MgSO^, filtered and the solvent evaporated. Chromatography on silica gel eluting with ethyl acetate gave the desired product, 40.7mg, mp. 293-295°C.

Example 6

Preparation of 2.4-Diamino-5-(2.3.5-trichlorophenv1)-6-methyl pyrimidine

1. Preparation of 2-(2,3.5-trichloropheny1)-3-oxobutyronitrile

To a solution of NaOEt (from 0.68g of sodium) in ethanol (20ml) was added 2,3,5-trichlorophenylacetonitrile (5g) and ethyl acetate (4.43ml). The mixture was heated under reflux, under nitrogen for 2.5 hours. The mixture was left standing at room temperature overnight. The mixture was then concentrated and the residue was dissolved in water. The aqueous phase was washed with ether, acidified with concentrated H₂SO^ and extracted with ether. The extracts were bulked, dried (MgSO^) and evaporated, 2.59g, mp. 134-135°C.

2. Preparation of 2-(2,3.5-trichlorophenyl)-3-methoxybut-2enonitrile

To a solution of the ketone in ether (100ml) at room temperature was added diazomethane (from 5.43g Oiazald) in ether (50ml) and the resulting mixture was left to stand at room temperature overnight. The ether was then distilled off to leave the desired C^rodLCt. 2.35c.

MJWD/MS/PA1024/22nd November 1989

- 27 3. Preparation of 2.4-0iamino-5-(2.3.5-trichlorophenyl)-6-methylpyrimidine

Guanidine hydrochloride (3.87g) was added to a solution of sodium ethoxide (from l.Olg of sodium) in ethanol (80ml). The resulting white suspension was stirred at room temperature for 10 minutes and then added to a solution of the enol ether (Example 6.2) (5.60g) in ethanol (20ml). The resulting mixture was stirred at reflux for 8 hours under nitrogen. After cooling, the suspension was filtered, and the filtrate evaporated to dryness in vacuo. Chromatography on silica gel eluting with CHCl-j to 2% MeOH-CHClg gave the desired product which was triturated with ether and dried in vacuo. Yield 1.70g, mp. 236-238°C.

Exawl· 7

I

Preparation of 4-Amino-2-N-morpholino-5-(2,3,5-trichlorophenyl)-6trifluoromethylpyrimidine.

To a solution of NaOEt (from 0.144g of Na) in EtOH (12.5ml) was added morpholinoformamidine hydrobromide (1.08g) (Lancaster Synthesis). The resulting white suspension was stirred at room temperature for 10 minutes. A solution of the enol ether (0.85g) (Example 1.6) in EtOH (2.5ml) was added and the resulting mixture was stirred at reflux for 4.5 hours. After cooling, the suspension was filtered, and the filtrate was evaporated to dryness in vacuo. Chromatography on silica gel, eluting with CHC1₃, gave the desired product which was triturated with petroleum ether (b.p. 40-60°C) and dried in vacuo. 0.47g, mp.

177-181°C.

AP π π π 1β 4

MJWD/MS/PA1024/22nd November 1989

- 28 Exawl· 8

Preparation of 4-Amino-2(-N.N-diroethy1aroino)-5-(2,3.5-trichlorophenyl)-6-trifluoromethyl pyrimidine.

To a solution of NaOEt (from 0.144g of Na) in EtOH (12.5ml) was added 1,1-dimethylguanidine sulphate (1.4g), (Aldrich). The resulting white suspension was stirred at room temperature for 10 minutes. A solution of 2-(2,3,5-tri chlorophenyl )-4,4,4-trifluoro-3-methoxybut-2-enonitrile (see Example 1.6) (0.85g) in EtOH (2.5ml) was added and the resulting mixture was stirred at reflux for 4.5 hours. After cooling, the suspension was filtered, and the filtrate was evaporated to dryness in vacuo. Chromatography on silica gel, eluting with CHClg, gave the desired product, 0.61g, mp. 124-126°C.

Exawl· 9

Synthesis of 4-Amino-2-N-morpholino-5-(2.3.5-trichlorophenyl)pyrimidine

To a solution of NaOEt (from 0.21g of sodium) in ethanol (20ml) was added morpholinoformamidine hydrobromide (1.6g). After stirring for 10 minutes the adduct of Example 3.2 (lg) was added and the mixture was stirred at reflux for 4 hours. The mixture was left standing at room temperature overnight and then filtered. The filtrate was concentrated and the residue was purified by chromatography on SiO^ gel, eluting with CHClg to give the desired product, O.73g, mp. 168-170°C.

MJHD/MS/PA1024/22nd November 1989

- 29 Ex—pie 10

Synthesis of 4-Amino-2-(N,N-dimethyl amino)-5-(2,3,5-tr ichlorophenyl )pyrimidine

To a solution of NaOEt (from 0.21g of sodium) in ethanol (20ml) was added 1,1-dimethylguanidine sulphate (2.07g). After stirring for 10 minutes the adduct of Example 3.2 (lg) was added and the mixture was stirred at reflux for 4 hours. The mixture was left standing at room temperature overnight and then filtered. The filtrate was concentrated and thi residue was purified by chromatography on S1O2. eluting with CHCl^ to give the desired product, 0.69g, mp. 145-147°C.

Ex—pie 11

Synthesis of 4-Amino-6-methyl-2-(4-methylpiperazin-l-yl)-5-(2,3,5trichlorophenyl)pyrimidine

To a solution of NaOEt (from 0.16g of sodium) in ethanol (15ml) was added N-methylpiperazinoformamidine hydriodide (1.6g). After stirring for 10 minutes the enol ether of Example 6.2 (0.82g) in ethanol (5ml) was added and the mixture was stirred at reflux for 5 hours. The mixture was left standing at room temperature overnight and then filtered. The filtrate was concentrated and the residue was purified by chromatography on Si0₂ gel, eluting with CHC1₃ to 4% MeOH/CHCl^ to give the desired product, 0.31g, mp. 156-159°C.

APO00164

Ex—pie 12

Synthesis of 2,4-diamino-6-methyl-5-(2.3,5-trichloro-4-nitropheny1) pyrimidine

2,4-0iamino-5-methyl-5-(2,3,5-trichlorophenyl)pyrimid ine (0.152g) (Example 6) was dissolved in concentrated sulphuric acid and potassium nitrate (50mg) was added. The solution was st^;r^ed -at room bad ORIGINAL

MJWD/MS/PA1024/22nd November 1989

- 30 temperature overnight, then poured onto ice and basified with 0.880 ammonia. The product was extracted into ethyl acetate, bulked, dried (MgSO^) and evaporated. The residue was purified by chromatography on Si0₂ gel, eluting with EtOAc to 8% MeOH/EtOAc to give the desired product, 0.13g, mp. 313-315°C.

Exawl· 13 ,

Synthesis of 4-Amino-2-(N,N-dimethylamino)-6-methyl-5-(2.3,5-tri chlorophenyl)pyrim1dine

To a solution of NaOEt (from 0.16g of sodium) in ethanol (15ml) was added 1,1-dimethylguanidine sulphate (1.6g). After stirring for 10 minutes the enol ether of Example 6.2 (0.82g) in ethanol (5ml) was added and the mixture was stirred at reflux for 5 hours. The mixture was left standing at room temperature overnight and then filtered. The filtrate was concentrated and the residue was purified by chromatography on Si0₂ gel, eluting with CHClg to give the desired product, 54mg, mp. 151-153°C.

Exawl· 14

Synthesis of 2.4-Diamino-$-(2,3-dichlorophenyl)-6-trifluoromethyl pyrimidine

1. Preparation of 2,3-dichlorobenzyl alcohol

To a solution of 2,3-dichlorobenzaldehyde (Aldrich, 50gms) in alcohol (800mL) at room temperature was added NaBH^ (8.54gms) and the resulting mixture stirred for 1.5 hours. The reaction was quenched with water and the solvent evaporated in vacuo before partitioning the residue between CHClg and saturated NaHCOg solution. The organic phase was washed with brine, dried over MgSO^, filtered and the solvent evaporated in vacuo to leave a white solid, 48.38gms, mp. 37-87.5°C.

MJWD/MS/PA1024/22nd November 1989

J» ‘‘ '

- 31 2. Preparation of 2.3-dichlorobenzyl bromide

To a solution of the alcohol in benzene (500ml) under H? was added PBr^ (167.8gms), and the mixture stirred at 55-60°C for 3.5 hours. After cooling, the mixture was poured onto crushed ice (2L) and the benzene layer separated. The aqueous phase was washed with benzene (x3) and the combined benzene extracts washed with saturated NaHCOg solution and water, dried over MgSO^, filtered and the solvent evaporated to leave a brownish liquid which solidified on standing. 37.53gms, mp. 31-32°C.

3. Preparation o*f 2,3-dichlorophenylacetonitrile

The bromide was suspended in DMSO (155ml)/ water(105ml) at 0°C and KCN (20.24gms) added in portions. After stirring at 30-35°C for 2 hours, the suspension was diluted with water and extracted with Et₂O. The combined ether extracts were washed with water, dried over MgSO^, filtered and the solvent evaporated in vacuo to leave a white solid, 27.52gms, mp. 64-67°C.

4. Preparation of 2-(2,3-dichlorophenyl)-4,4.4-trifluoro-3-oxobutyronitri le

To a solution of NaOEt (from 1.48gms Na) in EtOH (25ml) at room temperature under N₂ was added the nitrile (lO.Ogms) followed by ethyl trifluoroacetate (9.3gms) and the mixture stirred'at reflux for 5 hours. After cooling, the solvent was removed in vacuo and the residue dissolved in water. The aqueous phase was washed with Et₂O (discarded), acidified with H₂S0₄ and extracted with Et₂0. The combined Et₂0 extracts were washed with water, dried over MgSO^, filtered and the solvent evaporated in vacuo to leave an oil. This was triturated with petroleum ether, and the solid filtered off and dried. 9.56gms, mp. 74-75°C.

MJWD/MS/PA1024/22nd November 1989

- 32 5. Preparation of 2-(2,3-dichlorophenyl)-4.4.4-trifluoro-3methoxybut-2-enonitri le

To a solution of the trifluoromethyl ketone in Et₂0 (90ml) at room temperature was added diazomethane (from 19.35gms Diazald) in Et₂0 (180ml), and the resulting mixture left to stand at room temperature overnight. Excess diazomethane was then removed in vacuo into AcOH, and the residue was dissolved in Et₂O, dried over MgSO^, filtered and the solvent evaporated in vacuo to leave a brownish solid, 6.44gms.

6. Preparation of 2,4-Diamino-5-(2.3-dichlorophenyl)-6-trif luoro methylpyrimidine

To a solution of the above enol ether in ethanol (37ml) was added guanidine hydrochloride (1.92gms) followed by a solution of NaOEt (from 540mgs of Na) in EtOH (90ml), and the resulting mixture stirred at reflux for 3 hours. After cooling, the suspension was filtered, and the filtrate evaporated to dryness jn. vacuo. Chromatography on silica gel eluting with CHCl^ to 2% MeOH-CHCl^ gave the desired product which was triturated with Et₂0 and dried in vacuo, 673mg, m.pt.218-9°C.

7. 2,4-Diamino-5-(2,3-dichlorophenyl)-6-trifluoromethylpyrimidine methanesulphonate

To a suspension of the free base (lOOmg) in ethanol was added methanesulphonic acid (30mg) and the resulting clear solution stirred at room temperature for 2 hours. The solution was evaporated to dryness, and the resulting solid triturated with ether, filtered, and dried in vacuo, 107mg, m.pt. 253-256°C.

8. 2,4-Diami no-5-(2,3-di chlorophenyl)-6-trifluoromethylpyrimidine hydrochloride

To a solution of the free base (150mg) in methanol was added ethereal hydrogen chloride. After stirring, the solvent was evaporated to dryness and the resulting solid triturated with ether, filtered, and dried in vacuo. 160mg, m.pt. 233-235^7.

BAD ORIGINAL

MJWD/MS/PA1024/22nd November 1989

- 33 ExtwU 15

Synthesis of 2,4-Di ami no-5-(2,3-dichlorophenyl)-6-methylpyrimidine

This compound was made in an analogous manner to the compound of Example 6 from 2,3-dichlorophenylaceonitrile, m.pt. 245-247°C.

Exmpl· 16

Synthesis of 2,4-Diamino-5-(2,3-dichlorophenyl)-6-methoxymethy1 pyrimidine

1. Preparation of 2-(2,3-dichlorophenyl)-4-methoxv-3-oxo-butyronitrile

To a stirred refluxing solution of NaOEt (from l,38gms Na) in EtOH (25ml) was added a mixture of ethyl methoxyacetate (8.85gms) and 2,3-dichlorophenylacetonitrile (Example 14.3) (9.3g) dissolved in DME (20ml) during 5 minutes. After 5 hours a precipitate had appeared (sodium salt of product). The mixture was cooled and filtered, the filtrate evaporated to dryness jn vacuo and the residue partitioned between ether and water (the ether phase was discarded). .The aqueous residue was acidified with 2N H₂S0₄ and extracted with ether (2x). The combined Et₂0 extracts were washed with water, dried over MgSO^, filtered and evaporated in vacuo to give a yellow solid (a). The sodium salt (above) was dissolved in water and the solution extracted with ether and discarded. The aqueous solution was acidified with 2N H₂SO₄ and extracted with ether. The ether extract was washed with water, dried over MgSO^, filtered and evaporated in vacuo to give a white solid (b).

The above products (a) and (b) were combined to give a yield of 10.4gms which was used without further purification. Single soot by TLC (19:1 CH^Cl?: MeOH) Rf 0.35.

V 9 I· 0 U OdV

MJWD/MS/PA1024/22nd November 1989 bad original

- 34 2. Preparation of 2-(2,3-dich lorophenyl)-3,4-dimethoxybut-2-enonitri Ie

To a stirred solution of the above nitrile (9.4gms) in ether was added in portions diazomethane (0.4 - 0.45M) in ether. Initially vigorous frothing occurred and after further addition no immediate reaction was produced. The mixture was left stirring at room temperature for 3 hours and evaporated in vacuo, into AcOH to give the enol ether.

3· Preparation of 2.4-Diamino-5-(2.3-dichlorophenyl)-6methoxymethylpyrimidine

To a solution of NaOEt (from 0.92gms Na) in EtOH (40ml) was added guanidine hydrochloride (3.44gms). A solution of the enol ether (above) in EtOH (30ml) was added and the mixture refluxed for ca. 3 hours. After cooling, the solvent was evaporated off in vacuo and the residue treated with 5N NaOH (ca. 50ml). The red solid was filtered off, dissolved in AcOH (ca. 20ml), diluted with water (40ml), treated with charcoal, and filtered. The filtrate (yellow solution) was made alkaline with 2N NaOH, and the white precipitate filtered off, dried and recrystallised from EtOH, 4.39gms, mp. 237-240°C.

Extol· 17

Preparation of 2,4-0iamino-5-(1-naphthyl )pyrimidine

To a solution of NaOEt (from 1.45gms Na) in ethanol (60ml) at room temperature under N^ was added 1-naphthylacetonitriIe (Aldrich, 10.02gms). After stirring for 10 minutes, ethyl formate (8.88gms) was added and the mixture stirred at reflux for 5 hours. The mixture was cooled, the solvent evaporated, and the residue triturated with Et^O before filtering and drying the solid in vacuo (6.86gms).

MJWD/MS/PA1024/22nd November 1989

- 35 The above solid was dissolved in DMF (45ml), methyliodide (5.4gms) added, and the reaction vessel sealed before stirring the contents at 40°C for 4 hours. The solvent was then removed in vacuo. and the residue partitioned between EtOAc and water. The organic phase was washed with water, dried over MgSO^, filtered, and the solvent evaporated in vacuo to leave the crude product as a viscous reddish oil (5.4gms).

To a solution of NaOEt (from 1.19gms of Na) in ethanol (80ml) was added guanidine hydrochloride (4.94gms). After stirring for a further 5 minutes, the ab*ove intermediate in ethanol was added and the resulting mixture stirred at reflux for 3.5 hours. After cooling, the solvent was removed in vacuo and the residue partitioned between CHCl^ and water. The organic phase was washed with water, dried over MgSO^, filtered and the solvent evaporated to leave a pale yellow solid. Chromatography on SiO£ eluting with CHCl^ to 4% CHClyMeOH, followed by recrystallisation from ethanol gave the desired product as a white solid. 2.82gms, mp.l71-3°C.

APO 0 0 16 4

Ex awl· 18

Preparation of 2.4-Diamino-5-(2.3-dichlorophenyl)-6-fluoromethyl pyrimidine

1. 2,4-Diamino-5-(2,3-dichlorophenyl)-6-(diethoxymethyl) .

pyrimidine

To a stirred refluxing solution of NaOEt (from 1.38g sodium) in ethanol (25ml) was added over 5 minutes a mixture of ethyl diethoxyacetate (13.21g; 75mmol) and (Example 14.3) 2,3dichlorophenylacetonitrile (9.3g; 50mmol) in dry dimethoxyethane (20ml). After 4 hours cooled and evaporated in vacuo. The residue was partitioned between water (100ml) and_ether (100ml), the ether phase discarded and the aaueous residue acidified with

BAD

MJWD/MS/PA1024/22nd November 1989

- 36 IN H^SO^. Extraction with CH^Cl? gave the acyl acetonitrile (13.47g), which was used without further purification.

To a stirred solution of the above acyl acetonitrile in ether (100ml), cooled on ice was added in portions a solution of diazomethane (ca. 3g) in ether. After 2 hours the solution was evaporated in vacuo to give the desired enol ether as an oil, which was used without further purification.

To a solution of NaOEt (from 1.4g sodium) in ethanol (50ml) was added guanidine hydrochloride (4.8g; 50mmo1). A solution of the above enol ether in ethanol (20ml) was added and the mixture refluxed for 4 hours cooled, and concentrated in vacuo to ca. 30ml and diluted with water to give a dark purple solid which was filtered, dissolved in washed with water, dried over MgSO^ and evaporated jn vacuo. The residue was triturated with ethanol (50ml) and filtered to give the desired product (8.4g) which was used without further purification, (mp. 214-217°C).

2. 2.4-Diamino-5-(2.3-dichlorophenvl)pyrimidine-6-carboxaldehyde

A mixture of the above acetal (7g) and 0.4M HC1 (150ml) was refluxed with stirring for 1 hour, cooled on ice and neutralised with 2M NaOH. The mixture was filtered, washed with water and dried in air to give the desired product (6.2g), which was used without further purification.

3. 2,4-Di ami no-5-(2.3-dichloropheny1)-6-hydroxyroethvlpyrimidine

To a stirred solution of the above aldehyde (2.8g; lOmmol) in a mixture of dimethoxyethane (15ml) and ethanol (15ml) was added in portions sodium borohydride (llOmg; 3nmo1). After 30 minutes the solution was treated with water (50ml) and a few drops of acetic acid added to destroy excess borohydride. Extracted with dichloromethane (2 x 50ml), washed with water and the extract was then dried over MgSO^. Evaporation of the solvent, in vacuo gave a pink solid, which was triturated with ether, filtered and dried

MJWD/MS/PA1024/22nd November 1989

- 37 (1.6g). Recrystallisation from methanol (50ml) gave the desired product as fine colourless crystals. 0.65g, m.p. 173-6°C.

4. 2,4-Di ami no-5-(2.3-dichlorophenyl)-6-f luoromethylpyrimidine

To a stirred suspension of 2,4-diamino-5-(2.3-dichlorophenyl)-5hydroxymethylpyrimidine (185mg; lmmol) in dry dichloromethane (25ml), under nitrogen at -70°C, was added dropwise diethylaminosulphur trifluoride (263μ1; 2mmol). The mixture was allowed to warm to 0°C and kept at this temperature for 4 hours. After coolingL to -70°C the mixture was quenched with aqueous sodium bicarbonate, extracted with dichloromethane (2 x 50ml), washed with saturated brine and dried (MgSO^). Concentration gave a colourless gum (0.2g). Chromatography on silica gel, eluting with 0.01:1:19 Et₃N:MeOH:CH₂Cl₂ gave the desired product which was triturated with CCl^ and dried in vacuo, lllmg, mp. 224-6°C.

AP 0 0 0 1 6 4

Exaapl· 19

2.4-Diamino-5-(2.3-dichloropheny1)-6-phenoxvmethylpyrimidine To a stirred solution of NaOEt (from 1.38g sodium), in ethanol (70ml) at reflux, was added over 10 minutes a mixture of 2,3-dichlorophenylacetonitrile (9.3g) and ethyl phenoxyacetate (13.5g) . in dry dimethoxyethane (50ml). After stirring at reflux for 3 hours the mixture was cooled, filtered and the solvents evaporated in vacuo. The residue was dissolved in water, washed with ether (discarded), acidified with 2N hydrochloric acid and extracted with di chloromethane. The combined extracts were washed with brine, dried (MgSO^) and evaporated _£n vacuo to leave a tan coloured solid (8g), which was used without further purification.

To a suspension of the crude acyl acetonitrile (8g) in ether (150ml) was added in portions an excess of a solution of djazomethane in eider. After stirring for ’ hour at room temperature the s~1ut^;^as

BAD ORIGINAL

MJWD/MS/PA1024/22nd November 1989

- 38 concentrated j_n vacuo to give the enol ether, which was used without further purification.

To a solution of sodium ethoxide (from 0.63g sodium) in ethanol (25ml) at room temperature was added guanidine hydrochloride (2.39g). After 15 minutes a solution of the above enol ether in ethanol (25ml) was added and the mixture stirred at reflux for 4 hours. After cooling the solvent was evaporated in vacuo. The residue was suspended in 2N NaOH (75ml), filtered, washed with water, dried in air and recrystallised from ethanol to give the desired product as a colourless solid. 3.82g, mp. 2U-213°C.

Exawle 20

2.4-0iamino-5-(2,3-dichlorophenyl)-6-methylthiomethy1pyrimidine

To a stirred solution of NaOEt (from 1.38g sodium), in ethanol (25ml) at reflux, was added over 5 minutes a mixture of 2,3-dichlorophenylacetonitrile (9.3g) and ethyl methylthioacetate (10.07g) in dry dimethoxyethane (20ml). After stirring at reflux for 5 hours the mixture was cooled on ice, acidified with acetic acid (5ml), poured into cold water and extracted with dichloromethane. The combined extracts were washed with water, dried (MgSO^) and concentrated to give a yellow oil which was used without further purification.

The crude acyl acetonitrile was heated under N₂ with triethyl orthoformate (40ml) at 140-150°C for 4 hours, distilling off low boiling material. After cooling, the mixture was concentrated j_n vacuo to leave a dark oil (15.2g). The oil was dissolved in ethanol (20ml) and added to a mixture of guanidine hydrochloride (4.8g) and sodium ethoxide (from 1.38g sodium) in ethanol (50ml). After stirring at reflux for 4 hours the mixture was cooled and concentrated in vacuo. The residue was shaken with 5M NaOH to give a dark oil, which -as extracted with dichloromethane, washed with water and dried

MJWD/MS/PA1024/22nd November 1989

- 39 (MgSO^). Concentration in vacuo left a dark gum. The desired product crystallised from ethanol (20ml) as a light tan solid, 0.57g, mp. 205-7°C.

Example 21A

Preparation of 2,4-Diamino-5-(2.3-dichlorophenv1)pyrimidine

a. 2-(2,3-dichlorophenyl)-3-oxo-prop ionitriIe

To a solution of NaOEt (from 3.63gms Na) in ethanol (500ml) was added 2,3-dichlorophenylacetonitri le (Example 14.3) in ethanol (150ml). Ethyl formate (16.67gms) was then added and the mixture stirred at 80°C for 45 minutes, before adding a further quantity of ethyl formate (2.78gms). After stirring at 80°C for a further 1.5 hours, the precipitate was filtered off and dried j_n vacuo. The solid was dissolved in water , filtered, acidified with concentrated hydrochloric acid and the precipitate filtered off and dried in vacuo. 14.35gms, 45% yield.

b. A solution of the above product, ethylene glycol (9.19gms) and £-toluene-sulphonic acid (8.9gms) in toluene (100ml) was stirred at reflux, with the water being collected in a Dean and Stark trap. After cooling, the solution was washed with water, IN NaOH and water before drying over MgSO^. Evaporation of the solvent left an oil (20.17gms) which was dissolved in EtOH (70ml). After standing for 1 hour the cream precipitate was filtered off and dried in vacuo (10.44gms).

c. To a solution of NaOMe (5.lgms) in ethanol (75ml) was added guanidine hydrochloride (8.2gms). After stirring for 30 minutes the NaCl was filtered off, the above acetal added and the mixture stirred at reflux for 1 hour. The solvent was then concentrated and the product filtered o’<. Recrystallisation from ethano· >9lOOOdV

MJ'wC, MS, ?AiG24/22na Novemcer 1989 — bad ORIGIN*¹V

- 40 gave the product as a white solid, 7.67gms, 70% yield, mp.

212.5-214°C.

The diamino-pyrimidine (6.12gms) was dissolved in ethanol (250ml), concentrated hydrochloric acid (2.07ml) added, and the suspension chilled for 2 hours. The precipitate was then filtered off and dried to give the hydrochloride salt, 5.52gms.

Exawl· 21B

a. 2,3-Dichlorophenylacetic acid

Concentrated hydrochloric acid (100ml) was poured onto crushed ice (150ml), the solution added to 2,3-dichlorophenylacetonitrile (30.6gms), and the mixture refluxed for 3 hours. After cooling, the mixture was diluted with water (500ml), extracted with EtOAc (600ml) and the organic phase washed with brine before drying over MgSO^. Evaporation of the solvent left a white solid, 31.3gms.

b. Ethyl 2,3-dichlorophenylacetate

To a suspension of the acid in ethanol (200ml) was added concentrated H^SO^ (1ml) and the mixture stirred at reflux for 3 hours. After cooling, the solvent was evaporated and the residue treated with concentrated NH^OH (3ml) in water (50ml). The organic phase was extracted into C^Cl?, dried over MgSO^, and the solvent evaporated to leave a clear liquid, 19.88gms.

c. Ethyl-2-(2,3-dichlorophenvl)-3-N-morpho1ino-aerylate

To a mixture of the ester, morpholine (40.7gms) and ethyl orthoformate (69.24gms) was added acetic anhydride (0.5ml) and the resulting pale yellow solution stirred at reflux for 3 hours. After cooling, the mixture was concentrated in vacuo. A white precipitate started to form, and this was filtered off before

MJriD/MS;rAlC24_z'22na November 1989

BAD ORIGINAL further concentrating the filtrate to give a brownish clear oil. Standing overnight in vacuo gave a yellow solid, 34.34gms.

d. 5-(2,3-dichloropheny1) isocytosine

To the above ester was added guanidine hydrochloride (26.6gms) slurried in sodium 2-methoxyethoxide (from 6.6gms of Na) in 2-methoxyethanol (150ml), and the mixture stirred at reflux overnight. After cooling, the mixture was concentrated in vacuo, diluted with water (100ml) and then washed with Et^O (200ml). The aqueous phase was acidified with AcOH, and the precipitate filtered off and washed with EtOH and then Et^O before prying in vacuo, 13.48gms.

e. N^-fA-chloro-5-(2,3-dichlorophenyl )-2-pyrimidiny11-N²,N²dimethylformamidine

To a mixture of the above isocytosine (14.4gms) in CH₂C1₂ (200ml) was added dropwise during 30 minutes fresh Vilsmeier-Haack reagent (from 2.75 equivalents of SOC1₂ and 2.58 equivalents of DMF), and the mixture refluxed for 6 hours. After cooling, IN NaOH (250ml) was added slowly. The aqueous phase was washed with CH₂C1₂ and the combined organic extracts washed with brine before drying over MgSO^. Evaporation of the solvent and chromatography on Si0₂ gel, eluting with EtOAc gave 13.6gms, mp. 113-115°C.

f. 2-Amino-4-ch1oro-5-(2,3-dichlorophenyl )pyrimidine

To the formamidine in EtOH (50ml) was added ethanolic-MeNH₂ (8 equiv.) in EtOH (50ml), and the mixture sealed in a Parr reaction vessel before stirring at room temperature for 5 hours. The reaction mixture was then concentrated in vacuo, the residue mixed with IN NaOH (75ml), filtered, washed with water and dried in vacuo. 11.2gms, mp. 228-30°C.

5· 2,4-Oiamino-5-(2,3-dichlorophenyl) pyrimidine

To 2-amino-4-chloro-5-(2,3-dichlorophenyl)pyrimidine (6.73gms) was added ethanolic ammonia (30 equiv. in 50ml~Et0H) and the

V0'w3/MS/PA 1024/22nd November 1589

BAD ORIGINAL

APO00164

- 42 mixture sealed in a Parr reaction vessel and heated to 125°C for 38 hours. After cooling, the mixture was concentrated jn vacuo and the residue mixed with IN NaOH (75ml), filtered, washed with water and dried in vacuo. 6.14gms, mp. 208-211°C.

Example 22

Synthesis of 2,4-Diamino-5-(2-chlorophenyl)-6-methyl-pyrimidine

This compound was prepared according to JACS, (1951), 73. 3763-70, mp. 225°C.

Example 23

Synthesis of 2.4-Diamino-5-(2-chlorophenyl )pyrimidine

This compound was prepared according to JACS, (1951, 73. 3763-70, mp. 125-8°C.

Example 24

Synthesis of 2.4-0iamino-5-(2-chlorophenyl)-6-ethylpyrimidine Prepared as for Example 22, except that ethyl propionate replaced ethyl acetate, mp. 197-8°C.

Example 25

Synthesis of 2,4-Diami no-5-(2,4-dichlorophenyl)-6-trifluoromethyl pyrimidine

This compound was prepared in a manner analogous to the compound of Example 14 from 2,4-dichlorophenylacetonitri le (Aldrich): mp. 220.5221°C.

.^MJnC/MS/PA1024,^/22nd November 1989

BAD ORIGINAL

- 43 Exaaple 26

Synthesis of 6-Benzy 1 oxymethy1-2,4-diami no-5-(2,4-di chlorophenyl) pyrimidine

This compound was prepared in a manner analogous to the compound of Example 16 from 2,4-dichlorophenylacetonitrile and ethyl benzyloxyacetate, 3.77gms, mp. 171-172°C.

Example 27

2-(4-methylpiperazin-l-yl)-4-amino-5-(2,4-dichlorophenyl)pyrimidine

A) A solution of 55.7g (0.4equiv) of S-methylisothiourea sulphate in 280ml of water was prepared and gently heated on a steam bath with stirring. Then 40g (0.4mol) of N-methyl-piperazine was slowly dripped into the solution while sweeping the flask out with nitrogen. The evolved gases were collected in several portions of a solution of 132g of mercuric chloride in 400ml of ethanol, which caused evolved methylmercaptan to be precipitated as methylmercuric chloride. After the addition of the N-methylpiperazine was complete, the reaction was continued until no more methylmercuric chloride precipitated. The reaction mixture was then concentrated in vacuo and chilled which caused the N-methyl-Ν'-amidinopiperazine sulphate to crystallise; 50.79g was collected.

8) A mixture of 76.3g (0.356 mol) of o-formyl-2,4-dichlorophenylacetonitrile, 63.7g of isoamyl alcohol, 0.36g of £-toluenesulphonic acid, 895 ml of toluene, and 10 drops of concentrated sulphuric acid were heated under reflux for 20 hours in the presence of a Dean and Stark trap to remove water formed in the reaction. Then an equal portion of i-amyl alcohol and a few drops of sulphuric acid were added, and the reaction was 'MS,'^οΑ1024/Ζ2ηα .‘to/emcer 1989

BAD ORIGINAL heated for another 20 hours, until the theoretical amount of water had been collected. The solution was cooled.

C) An 8.2g portion of sodium was dissolved in 500ml of absolute ethanol, and 50g of N-methyl-N'-amidinopiperazine sulphate was added. The mixture was allowed to stir for 10 minutes. This was then added to solution 8. The mixture was refluxed with stirring for 6 hours, allowed to stand overnight and the solvent removed in vacuo. The residue was then extracted with dilute hydrochloric acid, which dissolved most of it. The solution was extracted three times with ether, followed by neutralization of the aqueous fraction, which precipitated a gum which solidified upon standing overnight; weight 30g. This was crystallised repeatedly from 50% ethanol with the aid of decolourising charcoal. Very slow cooling was required in order for crystals

to be	formed; mp. 137°C.
Anal.	Calcd for C^H	17C12N5:
	C, 53.27;	H, 5.07;	N, 20.71;
Found:	: C, 53.58;	H, 5.14;	N, 20.40.

Example 28

Synthesis of 2,4-Diami no-5-(2,5-dichlorophenyl )-6-trifluoromethyl pyrimidine

This compound was prepared in a manner analogous to the compound in Example 14 from 2,5-dichlorobenzyl alcohol (Lancaster Synthesis, 48.26g) to give the title compound in a yield of 3.85gms, mp. 215-217°C.

024/22nd November 1989

BAD ORIGINAL

- 45 Exawl· 29

Preparation of 2,4-Diamino-5-(2,3,5-trichlorophenyl) pyrimidine

Guanidine hydrochloride (3.20g) was added to a solution of sodium ethoxide (from 848mg sodium) in ethanol (52ml). The resulting white suspension was stirred at room temperature for 10 minutes. The enol ether from Example 3.2 (4.40g) was added and the resulting mixture stirred at reflux for 3.5 hours. After cooling, the suspension was filtered, and the filtrate evaporated to dryness in vacuo. Chromatography on silica gel eluting with CHClg to 3¾ MeOH-CHClg gave the desired product which was triturated with ether and dried in vacuo. Yield = 2.01g, mp. 246-249°C.

Ex awl· 30

Synthesis of 4-Amino-5-(3-bromophenvl)-6-methyl-2-(4-methylpiperazin

-1-yl) pyrimidine

To a solution of NaOEt (from 0.92g of sodium) in ethanol (75ml) was added 3-bromophenylacetonitrile (Aldrich, 7.85g) and ethyl acetate (3.52g). The mixture was heated under reflux for 6 hours. After cooling, the mixture was concentrated and the residue was dissolved in water. The aqueous phase was washed with ether, acidified with 2N HC1 and extracted with ether. The extracts were bulked, dried (MgSO^) and evaporated, 3.8g, mp. 97-103°C.

The resulting ketone (3.7g), ethylene glycol (5ml) and £-toluenesulphonic acid (lOOmg) were heated under reflux in toluene (100ml) in a Dean & Stark apparatus for 3.5 hours. The mixture was cooled, concentrated and water was added to the residue. The product was extracted with ether and the extracts were bulked, dried (MgSO^) and evaporated, 4.03g, mp. 63-71°C.

e^cer 1989

BAD ORIGINAL

- 46 To a solution of NaOEt (from 0.28g of sodium) in ethanol (30ml) was added N-methylpiperazinoformamidine hydriodide (2.7g). After stirring for 10 minutes, the ketal (1.41g) was added and the mixture was stirred at reflux for 4 hours. After cooling the suspension was filtered, and the filtrate was concentrated. The residue was purified by chromatography on SiO^ gel. eluting with 10% MeOH/CHClg to give the desired product, 0.48g, mp. 120-122°C.

Ex—ole 31

Synthesis of 2.4-Diamino-5-(l-naphthyl)-6-trifluoromethylpyrimidine

This compound was prepared in a manner analogous to the compound in Example 14 from 1-naphthylacetonitrile (Aldrich, lOgms), to give the title compound in a yield of 0.69gms, mp. 224-226°C.

Exawle 32

Preparation of 2-Amino-5-(2,4-dichlorophenyl)-4,6-dichloropyrimidine

1. Ethyl 2,4-dichlorophenylacetate

2,4-Dichlorophenylacetonitrile (27.9g, 150m.mol) was suspended in 2N NaOH (400ml) and the mixture refluxed for 4 hours. The cooled reaction mixture was extracted with ether (2x200ml) acidified to pH3 and the solid filtered and dried (22g, 70%).

The product (20g) was dissolved in EtOH (300ml) and concentrated H^SO^ (5ml) added carefully. The mixture was refluxed for 7 hours. The cooled reaction mixture was evaporated under reduced pressure and the residue partitioned between CH^Clg and water (30ml each). The organic layer was extracted with saturated NaHCOg solution (200ml), washed with water (100ml), dried and

HJnD/MS/PAiO24/22nG Novemner 1389

- 47 evaporated in vacuo to give ethyl 2,4-dichlorophenylacetate as an oil (22.2g, 89.5%).

2. Diethyl 2,4-DichlorophenYl malonate

Sodium (1.86g, 0.081M) was added in portions to absolute ethanol (150ml) with stirring. After all the sodium had dissolved a solution of ethyl 2,4-dichlorophenylacetate (20g) in diethyl carbonate (50ml) was added dropwise. The reaction mixture was heated until EtOH distilled over. The rate of addition was controlled such that it equalled the rate of distillation. After the completioh of addition the reaction mixture was heated and distilled for further 4 hours. The cooled reaction mixture was partitioned between water (300ml) and EtOAc (300ml) and the organic layer dried and evaporated in vacuo to give a yellow oil (21g, 85%).

3. 2-Ami no-5-(2,4-dichioropheny1)-4,6-dihydroxypyrimidine

Sodium (4.52g, 0.196M) was added in portions to ethanol (150ml). After all the sodium had dissolved guanidine hydrochloride (12.44g, 0.13M) was added followed by diethyl 2,4-dichlorophenyl malonate(20g, 0.0655M). The mixture was refluxed for 6 hours, EtOH was removed under reduced pressure and the residue partitioned between 2N NaOH (400ml) and EtOAc (400ml). The aqueous layer was acidified with concentrated hydrochloric acid with cooling and the precipitated solid was filtered and dried (llg. 62%).

4. 2-Amino-5-(2,4-dich)oropheny))-4,6-dichloropyrimidine

A mixture of 2-amino-5-(2,4-dich)orophenyl)-4,6-dihydroxypyrimidine (lOg), phosphoryl chloride (100ml) and dimethylani1ine (1.5ml) was refluxed for 6 hours. The cooled reaction mixture was carefully added to crushed ice and the insoluble solid was filtered and washed with 2N HCl, followed by water. The solid was resuspended in water, neutralised (0.88 NH^OH) with cooling, and the mixture stood overnight at room temperature. The

APO00164

MJWD/MS/PA1024/22nd November 1989

- 48 insoluble solid was filtered, dried and purified by flash column chromatography to give the title compound (2.5g, 22¾), mp.

211-213°C. Microanalysis:

Calcd; C, 38.83; H, 1.6; N, 13.59;

Found: C, 38.59; H, 1.53; N, 13.40.

Exawl· 33

Synthesis of 2,4-Diamino-6-chloro-5-(2,4-dich1orophenyl)-pvrimidine

A mixture of 2-amino-5-(2,4-dichlorophenyl )-4,6-dichloro'pyrimidine (0.5g) (Example 32) EtOH saturated with ammonia (20ml) and copper powder (0.05g) was heated in an autoclave at 180°C for 18 hours. The cooled reaction mixture was filtered, evaporated and the residue purified by flash column chromatography to give the title compound (0.12, 25¾). mp. 219°C. Microanalysis:

Calcd: C, 40.82; H, 2.55; N, 19.05;

Found: C, 41.27; H, 2.46; N, 18.74.

Exaapl· 34

Synthesis of 2-Ami no-4-ch loro-5-(2,4-d i chlorophenyl ).-6-methylthio pyrimidine

A mixture of 2-amino-5-(2,4-dichlorophenyl)-4,6-dichloropyrimidine (0.5g) (Example 32) THF (15ml), methanethiol sodium salt (0.113g), copper powder (0.05g) and tris[2-(2-methoxyethoxy)ethyljamine (0.1a) was neated in an autoclave at 180°C for 18 hours. The cooled reaction mixture was filtered, evaporated and the residue purified by flash column chromatography to give the title compound (0.262g, 52%), mp. 201-202°C (softens at 196°C). Microanalysis:

Calcd: c, 41.19; H, 2.50; N, 13.10;

F:und: C, 41.10; H, 2.52: N, 12.77.

MJWD/MS/PA1024/22nd November 1989

BAD ORIGINAL

- 49 Exampj· J§

Synthesis of 2,4-0iamino-5-(2,4-dichlorophenyl)-6-methy1th iopyrimidine

A mixture of 2-amino-4-chloro-5-(2,4-dichlorophenyl)-6-methylthio pyrimidine (0.5g) (Example 32) EtOH saturated with ammonia (20ml), copper powder (0.05g) and tris[2-(2-methoxyethoxy)ethyljamipe (.Olg) was heated in an autoclave at 180°C for 18 hours. The cooled reaction mixture was filtered, evaporated and the residue purified by flash column chromatography to give the title compound (Q.llg, 23.5%), mp. 191-192°C. Microanalysis:

Calcd: for 0.2 hydrate: C, 43.33; H, 3.41; N, 18.38;

Found: C, 43.37; H, 3.23; N, 18.33.

Exaapl· 36

4-Amino-5-(3.5-dichlorophenvl)-6-methyl-2-(4-methylpiperazin-l-y1) pyrimidine

a. 3,5-Dichlorophenylacetonitrile

A mixture of 3,5-dichlorobenzyl alcohol (Aldrich, 25g), thionyl chloride (100ml) and DMF (0.5ml) was stirred and refluxed for 4 hours. After cooling the mixture was concentrated in vacuo, the residue was taken up in ether, washed with saturated aqueous NaHCO^ and brine, dried (MgSO^) and concentrated in vacuo to give

3,5-dichlorobenzyl chloride as a light yellow solid, which was used without further purification, 28g, mp. 32-36°C.

To a vigorously stirred solution of 3,5-dichlorobenzylchloride (28g) in dichloromethane (150ml) was added a mixture of KCN (27.5g) and tetrabutylammonium hydrogen sulphate (2.38g) in water (110ml). After stirring at room temperature for 22 hours, the mixture was diluted with dichloromethane, the organic phase washed with water and concentrated in vacuo to 'eave an o^;'.

APO 0016 4 bad ORIGINAL

MJWD/M$/PA1024/22nd November 1989

- 50 Filtration through silica with toluene followed by concentration then trituration with hexane gave the desired product as a colourless solid. 15.8g, mp. 31-32°C.

b. 4-Amino-5-(3,5-dichlorophenyl)-6-methyl-2-(4-methylpiperazin-lvl)pyrimidine

To a stirred solution of NaOEt (from 0.59g sodium), in ethanol (25ml) at reflux, was added over 5 minutes a mixture of

3,5-dichlorophenylacetonitrile (9.3g) and ethyl acetate (3.3g) in dry dimethoxyethane (10ml). After stirring at reflux for 4 hours, the mixture was cooled on ice, acidified with acetic acid, poured into cold water and extracted with dichloromethane. The combined extracts were washed with water and concentrated to give an oil. Trituration with hexane gave 2-(3,5-dichlorophenyl)-3-oxobutyronitrile as a colourless solid (4.15g).

To a solution of the acyl acetonitrile (4.1g) in ether (100ml) was added in portions an excess of a solution of diazomethane in ether. After stirring for 2 hours at room temperature the solution was concentrated in vacuo to give the enol ether.

To a stirred solution of NaOEt (from 0.72g sodium) in ethanol (25ml) was added N-methylpiperazinoformamidine hydriodide (7.29g). After 10 minutes a solution of the above enol ether in ethanol (25ml) was added and then stirred and refluxed for 4.5 hours. After cooling the solvent was evaporated in vacuo and the residue shaken with 2N NaOH (50ml). The solid was filtered off, washed with water, dried in air and chromatographed (sijica; 1:9 MeOH.'CHCl^) to give the desired product as a colourless solid, 1.6g, mp. 154-166°C.

MJrtD/MS/PA1024/22nd November 1989

Exawl· 37

Preparation of 2.4-Diami no-5-(2,5-dichlorophenyl)-6-methylpyrimidine

This compound was made in an analogous manner to the compound of Example 6 from 2,5-dichlorobenzylalcohol (Lancaster Synthesis). Mp. 226-228°C: TLC (Si0_£: CHClj/MeOH, 9:1) Rf = 0.24.

Exawl· 38

Preparation of 2,4.-Diamino-5-(3.4-dich1orophenyl)-6-trifluororoethylpyrimidine

This compound was made in an analogous manner to the compound of Example 4, from 3,4-dichlorophenylacetonitrile (Aldrich) mp. 252-254.5°C: TLC (Si02i methanol/chloroform, 1:9) Rf = 0.38.

Exawl· 39

Preparation of 2,4-0iami no-5-(2,3-dichlorophenyl-4-nitrophenyl)pyrimidine

This compound was made in an analogous manner to the compound of Example 5 from 2,4-diamino-5~(2,3-dichlorophenyl)pyrimidine (Example 21). The reaction gave a mixture of the 4-nitro and 5-nitro derivatives from which the title compound was separated by column chromatography (Si02< EtOAc), mp. 237-9°C. Also separated in this manner was 2_r4-diamino-5-(2,3-dichloro-5-nitrophenyl)pyrimidine, mp. 264-6°C.

MJWD,MS/?A1024/22nd Novemoer 1989

- 52 Exaapl· 40

Preparation of 2,4-Diamino-5-( 2,4-dichlorophenyl )-6-(diethoxymethv 1) pyrimidine

This compound was prepared in a manner analogous to 2,4-diamino-5-(2, 3-dichlorophenyl)-6-(diethoxymethyl)pyrimidine (Example 18.1) from

2,4-dichlorophenylacetonitrile, mp. 225°C.

Exawl· 41

Preparation of 2,4-Diamino-5-(3,5-dichlorophenyl)-6-methylpyrimidine

This compound was prepared in a manner analogous to the compound of Example 6 from 3,5-dichlorophenylacetonitrile (Aldrich). mp. 242-244°C.

Exawl· <2

Preparation of 2,4-Diamino-5-(2,3-dichlorophenyl)-6-trifluoromethyl pyrimidine N-oxide

This compound was made from the compound of Example 14 by reaction with MCPBA in CHC1at room temperature. Mp. 275-278°C.

Exawl· 43

Preparation of 2,4-Diamino-5-(2,3-dichlorophenyl)-6-tri bromomethyl pyrimidine

This compound was prepared from the compound of Example 15 by reaction with excess bromine and sodium acetate in acetic acid at reflux. The title compound was separated from a mixture with the compound cf Example 75 by coloumn chromatography, mp. 210°C (dec)._

MJWD/MS/PA1024/22nd November 1989

Ex—pie 44

Preparation of 2.4-Diamino-5-(2,4-dich1orophenyl)-6-methox methyl pyrimidine

This compound was prepared in a manner analogous to Example 2, from

2,4-dichlorophenylacetonitrile, mp. 183-185°C. Single spot on TLC.

Ex—pie 45

Preparation of 2.4-0iamino-5-(2.6-dichlorophenyl)-6-methylpyrimidine

This compound was prepared in a manner analogous to Example 6 from

2,5-dichlorophenylacetonitrile (Aldrich), mp. 250°C.

Ex—pie 46

Preparation_of_2,4-Di ami no-5-(2,4-dichlorophenyl )pyr imid ine

-6-carboxaldehyde

This compound was made from the compound of Example 40 in an analogous manner to the compound of Example 18.2, mp. >350°C.

APO 0016 4

Ex—pie 47

Preparation of 2,4-Diami no-5-(2,3-dichloro-4-nitrophenyl)-6-methyl pyrimidine

This compound was made from the compound of Example 15 in an analogous manner to the compound of Example 39, mp. 265°C. Also obtained from this reaction was 2,4-diamino-5(2,3-dichloro-5-nitrophenyl)-6-methyl pyrimidine.

MJWD/MS/PA1024/22nd November 1989

- 54 Example 48

Preparation of 2.4-0iamino-5-(2.4-dichlorophenyl)-6-hydroxyimino methylpyrimidine

This compound was made from the compound of Example 46 by reaction with hydroxylamine hydrochloride in ethanol, mp. 260-5°C.

Exawle 49

Preparation of 2.4-Diami no-5-(2,4-dichlorophenyl)-6-hydroxymethyl pyrimidine

This compound was made from the compound of Example 46 in an analogous manner to the compound in Example 18.3, mp. 169-171°C.

Exawl e 50

Preparation of_2,4-Diamino-5-(2,3,4-trichlorophenyl)-6-methyl pyrimidine

This compound was made from the compound of Example 47 by reduction to the amine (PtO^, H_2> AcOH), formation of the diazonium salt (NaNO^, H^SO^), and reaction of this with CuCl (as for Example 57). Sublimes at 275°C. Homogenous on TLC (methanol/chloroform, 1:9) Rf=0.36.

Example 51

2,4-Diami no-5-(2,6-dichlorophenyl)-6-methoxymethy1-pyrimidine This compound was prepared in an analogous manner to the compound of Example 2 from 2,6-dichlorophenylacetonitrile (Aldrich), mp. 204-207°C.

MJWD/MS/PA1024/22nd November 1989

- 55 Exaapl· 52

Preparation of 2,4-Diami no-5-(2,3,5-trichloropheny 1 )-6-tri chloro methyl pyrimidine

This compound was made from the compound of Example 6 by reacting with NCS in AcOH at 100°C (AIBN as catalyst), mp. 226-227°C.

Exawl· S3

2,4-Di amino-5-(2,4~-d ich lorophenyl )-6-f luoromethylpyr imi dine

1. 2,4-0iamino-6-bromomethyl-5-(2,4-d ich lorophenyl )pyr imid-ine

2,4-0iami no-6-benzyloxymethy1-5-(2,4-dichioropheny1) pyrimidine (Example 26) (6.5g) was dissolved in a 47% solution of hydrobromic acid in acetic acid (75ml) and the mixture stirred and heated at 100°C for 6 hours. After standing at room temperature overnight the di hydrobromide · salt was filtered off, washed with ether and dried in vacuo. 6g.

To a stirred solution of the dihydrobromide salt (0.43g) in dimethylsulphoxide (4ml) was added dropwise a solution of sodium bicarbonate (0.84g) in water (10ml). After 30 minutes the precipitate was filtered off, washed with water then ether and dried in vacuo, 0.26g, mp'.>270°C (decomposes).

2. 2,4-0iamino-5-(2.4-dichloropheny1)-6-fluoromethylpyrimidine

To a solution of 2,4-diamino-6-bromomethyl-5-(2,4-dichlorophenyl) pyrimidine (1.04g) in tetramethylene sulphone (4.5ml) was added cesium fluoride (lg). The mixture was stirred and heated at 100°C for 4 hours, cooled, diluted with water and extracted with chloroform. The combined extracts were washed with water, dried (MgSO^) and concentrated in vacuo. The residue was chromatographed (silica; 19:1:0.1 dichloromethane/methanol/ triethylamine) to give the title compound_, which was recrysta1¹ised from ethanol. 0.19g, mp. 210-211°C.

y 9 I· 0 0 OdV

MJWD/MS/PA1024/22nd November 1989

- 56 Εχ«ρ1· 54

Preparation of 2.4-Diamino-5-r2-chloro-5-(N,N-dimethylsulphamoyl) phenyl]-6-methylpyrimidine

1. 2_l4-Diamino-5-(2-chloro-5-nitrophenyl)-6-methylpyrimidine

To a solution of 2,4-diamino-5-(2-chlorophenyl)-6-methyl pyrimidine (11.84g) (Example 22) in concentrated H₂S0₄ (100ml), was added potassium nitrate (5.1g). After stirring at room temperature for 90 minutes, the solution was poured onto ice and basified with ION NaOH. The product was extracted with ethyl acetate, bulked, dried (MgS0₄) and evaporated, 13.9g, 236-240°C.

2,4-Diami no-5-(5-amino-2-chloropheny1)-6-methy1pyrimidine A solution of 2,4-diamino-5-(2-chloro-5-nitrophenyl)-6-methyl pyrimidine (13.9g) in acetic acid (500ml) was reduced under an atmosphere of hydrogen in the presence .of PtO₂ (0.28g). The filtered through hyflo and the filtrate was

The residue was neutralised with saturated NaHCO-, mixture was concentrated solution and the product was extracted with ethyl* acetate, bulked, dried (MgSO^) and evaporated. Chromatography on Si0₂ gel, eluting with CHCK to product, 6g, mp. 117-121 C.

40% MeOH/CHClg, gave the desired

3. 2,4-Diamino-5-(2-chloro-5-N,N-dimethylsu1phamoylphenyl)-6-methyl pyrimidine

2,4-Di ami no-5-(5-ami no-2-chloropheny1)-6-methylpyrimidine (0.25g) was dissolved in water (0.8ml) and concentrated HC1 (0.5ml). To the cooled solution (below 10°C) was added a solution of sodium nitrite (0.07g) in water (0.5ml). After stirring at room temperature for 2 hours, the solution was cooled to 5°C. Cupric chloride (0.05g) and 5.14M S0₂ in acetic acid (0.97ml) were added and the reaction was stirred at 5°C overnight. The mixture was

MJ'wD/MS/PA1024/22na NovemDer 1989

- 57 filtered and washed with water to give the sulphonyl chloride, 0.23g.

The sulphonyl chloride (0.16g) was dissolved in THF (2ml) aqueous dimethylamine (2ml) was added. After stirring overnight, the solution was diluted with water, extracted with ethyl acetate, bulked, dried (MgSO^) and evaporated. Chromatography on Si0₂ gel, eluting with 2% MeOH/CHCl^, gave the desired product, 0.047g, mp. 283-285°C.

Example 55

Preparation of_2,4-Di ami no-(3,5-dichlorophenyl)-6-methoxymethyl pyrimidine

This compound was prepared in an analogous manner to the compound in Example 2 from 3,5-dichlorophenylacetonitrile, mp. 228-230°C.

Exawla 56

Preparation of 2.4-Diamino-5-(2,3-dich1orophenyl)-6-hydroxvpyrimidine

1. Ethyl-2-cyano-2-(2,3-dichlorophenyl)acetate

Sodium (1.2g) was added portionwise to ethanol (50ml) with stirring. After the sodium had dissolved a solution of

2,4-dichlorophenylacetonitrile (9.4g) in diethylcarbonate (25ml) was added dropwise. The reaction mixture was heated until EtOH distilled over. The rate of addition was controlled such that it equalled the rate of distillation. After the completion of the addition, the reaction mixture was heated and distilled for a further 4 hours. The cooled reaction mixture was partitioned between water and EtOAc (300ml each). The organic layer was dried and evaporated in vacuo and the residue vyas purified by flash column chromatography tc give the title product. (5g, 39%',.

MJWD/MS/PA1024/22nd November 1989

AP 0 0 0 1 6 4

BAD ORIGINAL

- 58 2.

2.4-Diami no-5-(2.3-dichlorophenyl)-6-hydroxypyrimidine

Sodium (1.2g, 0.052mol) was added in portions to absolute ethanol (50ml) with stirring. After the sodium had dissolved guanidine hydrochloride (3.69g, 0.039mol) was added followed by ethyl-2-cyano-2-(2,3-dich1orophenyl)acetate (5g, 0.0195mol). The mixture was refluxed for 8 hours, EtOH was removed under reduced pressure and the residue was partitioned between EtOAc and water. The EtOAc layer was extracted with 2N NaOH and the extract was neutralised with 2N HC1 with cooling. The precipitated solid was filtered and dried to give the title compound. (0.22g), mp. 275°C (decomp). Microanalysis:

Calcd: for 0.25 hydrate: C, 43.56; H, 3.09; N, 20.33;

C, 43.75; H, 3.09; N, 20.03.

ExawU 57

Synthesis of 2,4-0iamino-5-(2,4,5-trichlorophenyl)-6-methylpyrimidine

a. Preparation of 2-(2,4-dich1orophenyl)-3-oxo-butyronitri1e

A solution of 2,4-dichlorophenylacetonitrile (30.00gms, 161mmol) (Aldrich) in dry ethyl acetate (36ml) was added dropwise to an ethanolic solution of sodium ethoxide, prepared in situ from sodium metal (4.90g, 213mmol) and dry ethanol (60ml). This reaction mixture was refluxed for two hours, allowed-to stand overnight at room temperature and the ethanol was evaporated. The yellow solid obtained was dissolved in water and the resulting solution was extracted twice with ether. The aqueous layer was chilled and acidified with hydrochloric acid. The crude product was extracted with ether to give 23.31g of white solid.

MJWD/MS/PA1024/22nd November 1989

- 59 b. Preparation of 2.4-diamino-5-(2,4-dich1oropheny1)-6-methy1 pyrimidine

A solution of crude 2-(2,4-dichlorophenyl)-3-oxo-butyronitrile (23.24gs) in dry toluene (400ml) was refluxed with- ethylene glycol (280ml) and £-toluenesulphonic acid (8.00g, 42mmol) for four hours using a Dean and Stark trap. After cooling, the organic phase was washed with saturated NaHCOg, dried over MgSO^, and the solvent evaporated to leave a solid (24.0gm).

Finely ground guanidine hydrochloride (19.Ig, 200mmol) was added to an ethanolic solution of sodium ethoxide, prepared in situ from sodium metal (5.0g, 218mmol) in dry ethanol (500ml). A solution of the ketal (25.Og, 92mmol) in dry ethanol (10ml) was added to the guanidine solution. This mixture was refluxed for two hours and allowed to stand overnight at room temperature. The ethanol was evaporated and the crude product recrystal 1ised from hot acetone to give 17.23g of product, mp. 222-222.5°C.

c. Preparation of 2.4-Diamino-5-(2,4-dichloro-5-nitropheny1)-6methylpyrimidine

Finely ground potassium nitrate (6.5g 64mmol) was added to a solution of 2,4-diamino-5-(2,4-dichlorophenyl)-6-methyl pyrimidine (17.23g, 64mmol) in concentrated sulphuric acid (150ml). This mixture was stirred at room temperature for 90 minutes. The reaction mixture was then added to sodium bicarbonate and ice. The product was extracted with ethyl acetate. After the ethyl acetate was removed, a yellow solid was obtained (30.86g). A portion of this crude product (7.0g) was passed through a silica flash chromatography column and eluted with ethyl acetate to give the pure product (4.81g).

AP 0 0 0 1 6 4

MJWD/MS/PA1024/22nd November 1989

- 60 d. Preparation of 2.4-Diamino-5-(5-amino-2.4-dich1oropheny1 )-6methylpyrimidine

2,4-diami no-5-(2,4-dichloro-5-nitropheny1)-6-methylpyrimidine (4.80g, 15mmol) was dissolved in glacial acetic acid (18ml). This solution and lOmg of Adam's catalyst was stirred under an atmosphere of hydrogen at room temperature for 4 hours. The catalyst was filtered off and the acetic acid was evaporated. The colourless liquid obtained was dissolved in ethyl acetate and washed 3 times with water. After evaporating the ethyl acetate, a white solid was obtained (2.64g, 9mmol).

e. Preparation of 2,4-Diamino-5-(2.4,5-trich1orophenyl)-6-methyl pyrimidine

2,4-diami no-5-(5-ami no-2,4-dichlorophenyl)-6-methylpyrimidine . * .

(1.95g, 7mmol) was dissolved in a mixture of concentrated hydrochloric acid (3.6ml) and water (6ml). The temperature was lowered to 10°C. A chilled aqueous solution (3.6ml) of sodium nitrite (Q.50g, 7mmol) was added dropwise, keeping the temperature at 10°C. This mixture was stirred at room temperature for 2 hours, then chilled before adding it dropwise to a cold solution of cuprous chloride (1.7g, 17mmol) in concentrated hydrochloric acid (50ml). A grey solid precipitated which was filtered off and dried. This crude product (2.30g) was dissolved in ethyl acetate and washed twice with ammonium hydroxide solution and once with brine. After evaporating the ethyl acetate an off-white solid was obtained (2.04g). After recrystal 1isation from 10% methanol in chloroform the pure product was obtained (0.55g, 2mmol), mp. 262°C (dec).

MJWD/MS/PA1024/22nd November 1989

- 61 Example 53

Synthesis of 4-amino-2-(ethylamino)-5-(2.3.5-trichlorophenyl) pyrimidine

To a solution of NaOEt (from O.lg of sodium) in ethanol (10ml) was added ethylguanidine sulphate (lg) (Aldrich). After stirring for 10 minutes, the enol ether (Example 3.2) (0.486g) was added and the mixture was stirred at reflux for 4 hours. The reaction was left standing at room temperature overnight and then filtered. The filtrate was concentrated and the residue was purified by chromatography on SiO^ gel, eluting with CHCl^ to give the desired product, O.llg, mp. 149-152°C.

Exawle 59

2.4- Di ami no-5-(2,4-dichlorophenyl)-6-cyanomethyl-pyrimidine

This compound was prepared from 2,4-diamino-5-(2,4-dichlorophenyl)-6bromomethyl-pyrimidine (Example 53) by reaction with sodium cyanide in DMF at room temperature, mp. 249-251°C.

Example 60

2.4- 0iami no-5-(2.4-dichioropheny1)-6-dimethyl ami nomethyl pyrimidine

This compound was prepared from 2,4-diamino-5-(2,4-dichlOrophenyl)-6bromomethyl-pyrimidine (Example 53) by reaction with dimethylamine in ethanol at room temperature, mp. 170-172°C.

APO00164

Example 61

2,4-Diami no-5-(2,4-d ichlorophenyl)-6-cyanopyrimid ine

This compound was prepared from the compound of Example_48 by reaction with trifluorcacetic anhydride in pyridine, mp. 249°C.

MJ'nO/MS· ?-'.O24 ·22ηύ Novemoer 1969

- 62 Exawl· 62

2,4-Diamino-5-(2-chloro-4-fluorophenyl)-6-methylpyrimidine

This compound was prepared in an analogous manner to the compound of Example 15 from 2-chloro-4-fluorophenylacetonitrile, which was itself prepared from 2-chloro-4-fluorotoluene (Aldrich, mp. 238°C.

Ex awl· 63

2.4- Diami no-5-(3,4-di chiorophenyl)-6-methoxymethyl pyrimidine

This compound was prepared in an analogous manner to the compound of Example 2 from 3,4-dichlorophenylacetonitrile (Aldrich), mp. 204-206°C.

Exawl· 64

2.4- Diamino-5-(2.3-dichiorophenyl)-6-ethylpyrimidine

This compound was prepared in an analogous manner to the compound of Example 15 from ethyl propionate, mp. 228-230°C.

Exawl· 65

2.4- Diamino-5-(2, 4-d ifluoropheny1)-6-methyl pyrimidine

This compound was prepared in an analogous manner to the compound of Example 6 from 2,4-difluorophenylacetonitrile (Aldrich), mp. 291-296°C.

MJWD/MS/PA1024/22nd November 1989

- 63 Exawl· 66

2,4-Diami no-5-(2-naphthy))-6-methylpyrimidine

This compound was prepared in an analogous manner to the compound of Example 6 from 2-naphthylacetonitrile (Aldrich), mp. 221-222°C.

Ex awl a 67

2,4-Diami no-5-(1-naphthyl)-6-methy1pyrimidine

This compound was prepared in an analogous manner to the compound of Example 6 from 1-naphthylacetonitrile (Aldrich), mp. 224-225°C.

Exawl· SB

2-Hydroxv-4-amino-5-(2.3-dichlorophenyl) pyrimidine

This compound was prepared from the compound of Example 21 by reaction with sodium nitrite in IN H^SO^ at reflux to give a mixture of the title compound and the compound of Example 218.d. The title compound was separated by column chromatography, mp. 330-334°C.

AP 0 0 0 1 6 4

Exawl· 69

2-Amino-4-ethoxy-5-(2,4-dichlorophenyl)-6-methy1th iopyrimidine

This compound was made from the compound of Example 32.4 by reaction with methanethiol sodium salt in ethanol, mp. 123-124°C.

MJHD/MS/PA1024/22nd November 1989

- 64 Exawl· 70

2,4-Di ami no-5-(2,3,5-trichlorophenyl )-6-hydroxymethylpvrimidine

This compound was made from the compound of Example 2 by reaction with trimethylsilyl iodide in sulpholane at 80°C, mp. 101-105°C.

Exawl· 71

2,4-Di ami no-5-(2,3.5-trichlorophenyl)-6-fluoromethylpyrimidine This compound was .prepared in an analogous manner to the compound of Example 18 from 2,4-diamino-5-(2,3,5-trichlorophenyl)-6-hydroxymethylpyrimidine, mp. 215-217°C.

Exawl· 72

2,4-Di ami no-5-(2,4-di chlorophenyl )-6-carbamoylpyrimidine

This compound was prepared from the compound of Example 61 by reaction with concentrated sulphuric acid at room temperature, mp. 298-299°C.

Exawl· 73

2,4-Diamino-5-(2,4-dichlorophenyl)pyrimidine-6-carboxylic acid

This compound was prepared from the compound of Example 46 reaction with potassium permanganate, mp. 227°C.

Exawl· 74

Ethyl-2,4-diamino-5-(2,4-dichlorophenyl)pyrimidine-6-carboxvlate

This compound was prepared from the compound of Examp.le 73 by refluxing in ethanol in the presence of concentrated sulphuric acid, mo. 177.5°C.

MJ^D/MS/PA1024/22nd November 1989

- 65 Example 75

2,4-0iami no-5-(2,3-dichlorophenyl)-6-dibromomethylpyrimidine This compound was prepared from the compound of Example 15 by reaction with 2 eq NBS in CCl^ and AI8N as initiator. The title compound was separated from a mixture with the compound of Example 43 by column chromatography, mp. 270°C (dec).

Example 76

2-Dimethylami no-4-ami no-5-(2,4-dichloropheny1)pyrimi dine This compound was prepared in an analogous manner to the compound Example 10 from 2,4-dichlorophenylacetonitrile (Aldrich), mp. 151°C

Example 77

2-Dimethylamino-4-amino-5-(3,4-dichloropheny!)-6-methy1pyrimidine

This compound was made in an analogous manner to the compound Example 13 from 3,4-dichlorophenylacetonitrile.

Example 78

2-N-Piper idyl-4-amino-5( 2,4-dichloropheny]) pyrimidine This compound was prepared in an analogous manner to the compound Example 76 from 1-piperidinecarboxamidine sulphate (Bader), i 169°C.

of •<r <o <o o

o

D, of of

MJWD/MS/PA1024/22nd November 1989

- 66 Exawl· 79

2-Methylamino-4-amino-5-(2.3,5-trichlorophenyl)pyrimidine

This compound was prepared in an analogous manner to the compound of Example 58 from 1-methyl-guanidine hydrochloride (Aldrich), mp. 155-157°C.

Exawl· 80

2,4-Diami no-5-(2-chloro-5-bromophenyl)-6-methylpyrimidine

This compound was prepared in a similar manner to the compound of Example 54 by reaction of the diazonium salt with cuprous bromide, mp. 212-216°C.

Exawl· 81

2,4-Diami no-5-(2-chloro-5-iodophenyl)-6-methy1pyrimidine

This compound was prepared in a similar manner to the compound of Example 54 by reaction of the diazonium salt with potassium iodide, mp. 232-234°C.

Exawl· 82

2,4-Diamino-5-(2-chloro-5-cyanophenyl)-6-roethylpyrimidine

This compound was prepared in a similar manner to the compound of Example 54 by reaction of the diazonium salt with cuprous cyanide, mp. 239-241°C.

MJWD/MS/PA1024/22nd November 1989

- 67 Exawl· 83

2.4- Diamino-5-(2-chloro-5-fluorophenyl)-6-methylpyrimidine

This compound was prepared in a similar manner to the compound of Example 54 by way of the diazonium tetrafluoroborate salt, mp. 195-197°C.

Exawl· 84

2.4- Diamino-5-(2-chloro-5-methy1thiophenyl)-6-methylpyrimidine

This compound was prepared in a similar manner to the compound of Example 54 by reaction of the diazonium salt with methanethiol in the presence of copper powder, mp. 194-198°C.

Exawla 85

2-Amino-4,6-di(methyl thio )-5-(2.4-dichlorophenyl)pyrimidine

This compound was prepared from the compound of Example 32.4 by reaction with methanethiol sodium salt in methanol in the presence of tris[2-(2-methoxyethoxy)ethyl]amine and copper powder, mp. 164-165°C.

Exawl· 86

2,4-Diamino-5-(2-ch1oro-5-methanesu1phonylaminophenyl)-6-methyl pyrimidi ne

This compound was prepared from 2,4-diamino-5-(2-chloro-5-aminophenyl) -6-methylpyrimidine from Example 54 by reaction with methanesulphonylchloride in pyridine, mp. 234-240°C.

MJWD/MS/PA1024/22nd November 1989

- 68 Exawl· 87

2,4-Diamino-5-(2,3-dichlorophenyl)-l-methylpyrimidinium iodide This compound was prepared from the compound of Example 21 and methyl iodide, mp. 280-284°C.

Exawl· 88

2-Amino-4-methylamino-5-(2,3-dichlorophenyl)pyrimidine

This compound was prepared in an analogous manner to the compound of Example 21B.g by reaction with methylamine in ethanol, mp. 233-237°C.

Exawl· 89

2-Amino-4-dimethylamino-5-(2,3-dichlorophenyl)pyrimidine hydrochloride

This compound was prepared in an analogous manner to the compound of Example 21B.g by reaction with dimethylamine in ethanol and subsequent conversion to the hydrochloride salt, mp. 295-300°C.

Exawl· 90

2-Amino-4-chloro-5-(2,4-dichlorophenyl)pyrimidine

The compound was prepared in an analogous manner to he compound of Example 21B.f. from 5-(2,4-dichlorophenyl)isocytosine, mp. 2!5-216°C.

Exawl· 91

2-Amino-4-methylamino-5-(2,4-dichlorophenyl) pyrimidine

This compound was prepared from the compound of Example_ 90 by reaction «’tr methylamine in ethanol, me. 185-90 C.

MJWD/MS/PA1024/22nd November 1989

- 69 Exawl· 92

2-Amino-4-dimethy1 amino-5-(2,4-dichlorophenyl)pyrimidine hydrochloride

This compound was prepared from the compound of Example 90 by reaction with dimethylamine in ethanol and subsequent conversion to the hydrochloride salt, mp. 297-301°C.

Exawl· 93

2-Amino-4-piperidino-5-(2,4-dichlorophenyl)pyrimidine hydrochloride

This compound was prepared from the compound of Example 90 by reaction with piperidine in ethanol and subsequent conversation to the hydrochloride salt, mp. 303°C (dec).

APO00164

MJWD/MS/PA1024/22nd November 1989

- 70 Preferred among the compounds of formula (I) are the pyrimidines of the foregoing Examples 1,2,3,4,14 and 16, together with salts (in particular, pharmaceutically acceptable salts) thereof; these bases have the following respective two-dimensional structures.

Ex&apl· 1

Exaapl· 2

Example 3

MJWD/MS/PAlO24/22nd November 1989

Exaapl· 14

Ex«jqple 16

Ql <

MJWD/MS/PA1024/22nd November 1989

- 72 TABLE OF NMR DATA (g)

EXAMPLE MO. SOLVENT

ASSIGNMENT l· CDC1₃ 7.56(d,lH), 7.18(d,lH), 4.65-4.50 (br.s,2H), 3.88(t,4H), 2.5(t,4H), 2.36{s,3H)

DMSO-dg

DMSO-dg

3.06(s,3H,-0Me), 3.8(d,1H,J12.5Hz, -CH₂0Me), 3.9(d,1H,JI2.5HZ,-CH₂0Me), 5.98(br.s,2H,-NH₂), 6.1(br.s,2H,-NH^), 7.32(d,1H,J2.5Hz,6'-H), 7.78(d,1H,

J2.5Hz,4¹-H)

7.8(d,lH), 7.65(S,1H), 7.36(d,lH), 6.33-6.23(brs,2H), 3.68(t,4H), 2.32(t,4H), 2.2(s,3H)

DMSO-dg

6.40(s,2H), 6.55(s,2H), 7.35(s,lH), 7.80(s,lH)

DMSO-dg

8.6(s,lH), 7.49(s,lH), 6.4-6.3(br.s,2H), 6.25-6.15(br.s,2H)

OMSO-dg

1.70(s,3H), 7-30(s,1H),

5.75(s,2H), 5.90(s,2H), 7.75(s,lH)

COC1₃

7.55(d,1H),

18(d,lH), 4.75-4.58 (br.s,2H), 3.9-3.7(m,8H)

CDC1₃ . 55(d, 1H), 7.18(d, 1H), 4_.56-4.50 (b-.s.2H), 3.2(s,6H)

MJWD/MS/PA1024/22nd November 1989

- 73 10

DMSO-dg

7.79(d,lH), 7.67(s 6.47-6.27(br.s,2H)

1H), 7.36(d,lH), 3.72-3.57(m,8H)

DMSO-dg

7.78(d,lH), 7.64(s,lH), 7.35(d,lH), 3.08(s,6H) coci₃

7.51(d,1H) (br.s,2H). 2.34(s,3H)

7.17(d,1H)

3.82(t,4H),

2.0(s,3H)

4.40-4.22

2.48(t,4H),

DMSO-dg

8.28(s,lH), 6.18-6.04(br.d,4H), 2.1(s,3H)

COC1₃

7.51(d,lH), 7.18(d,lH) (br.s,2H), 3.16(s,6H),

4.36-4.22

2.0(s,3H)

DMSO d, b

6.10(s,2H), 6.45(s,2H) 7.3O(t,lH), 7.55(d,lH)

15(d,lH),

APO0016 4

DMSO-d_c

1.7O(s,3H), 5.60(s,2H), 7.15(d,lH), 7.3O(t,lH),

5.80(s,2H), 7.55(d;1H)

DMSO-dg 3.04(s,3H,-0Me),3.76(d,lH,J12Hz,-CH₂0Me)

3.85(d,1H,J12Hz,-0CH₂0Me), 5.84 (br.s,2H,-NH₂), 6.05(br.s,2H,-NH₂),

7.22(dd,1H,J7.5,1.5Hz,6'-H), 7.38 (dd,1H,J7.5Hz,5‘-H), 7.6(dd,lH,J7.5, 1.5Hz,4 ‘-H)

OMSO-dg

7.3-8.0(m,8H), 6.0-6.1(br.s,2H), 5.2-5.4(br.s,2H)

DMSO-d, b

4.75(2xdd,2H,J47,15Hz,-CH₂F), 5.95(br.s,2H,-NH₂), 6.15(_br. s,2H, -NH₂), 7.25(dd, IH.J7.5. 1.3Hz.5'-H), 7.39

MJWD/MS/PA1024/22nd November 1989

DMSO-dg (dd.lHJ7.5Hz), 7.64(dd,lH, J7.5, 1.0Hz)

4.43(d,1H,JllHz), 4,53(d,1H,J11Hz), 5.95(br.s,2H), 6.12(br.s,2H), 6.7(m,2H), 6.85(dd,1H,J7Hz),

7.l-7.4(m,4H), 7.55(dd,lH,J7,lHz).

DMSO-dg 4.4(d,lH,J12Hz,-CH₂0Ph), 4.52(d,lH,

J12Hz, -CH₂0Ph), 5.92(br.s,2H,-NH₂),

6.12(br.s,2H, -NH₂), 6.69(dd,1H,J7.5, 1.0Hz,6'-H), 6.85(dd,1H,J7.5Hz,5'-H), 7.10-7.35 (m,5H,-0Ph), 7.55(dd,1H,J7.5, 1.OHz,4'-H)

DMSO-dg 7.52(s,lH), 7.15-7.75(m,3H),

6.02(br.s,4H,2x-NH₂)

DMSO-dg 6.07(s,2H), 6.25(s,2H), 7.25(d,lH),

7.45(d,lH), 7.63(s,lH)

DMSO-dg 3.88(d,1H,JllHz), 4.0(d,1H,JllHz),

4.3(s,2H), 5.9(br.s,2H), 6.1(br.s,2H), 7.05-7.2(m,2H), 7.2-7.35(m,4H),

7.4(dd,1H,J8,2.5Hz), 7.62(d,1H,J2.5Hz)

DMSO-dg

6.25(s,2H), 6.50(s,2H), 7.30(s,lH), 7.40(d,lH), 7.5O(d,lH)

DMSO-dg

5.85(s,2H),

7.45(s_r1H),

6.1(s,2H), 7.25(s,lH), 7.7(s,1H)

COC1₃

7.53-7.12(m,4H), 4.48-4.30(br.s,2H), 3.81(t,4H), 2.46(t,4H), 2.33(s,3H),

2.03(s,3H)

MJWD/MS/PA1024/22nd November 1989

- 75 32

CDC1₃ 7.38(d,lH), 7.2(dd,lH), 7.08(d-,lH),

8.2(br.s,2H)

DMSO-dg 7.7(d,lH), 7.48(dd,lH), 7.29(d,lH),

6.45(br.s,2H), 6.2(br.s,2H)

CDC1₃ 7.5(d,lH), 7.35(dd,lH), 7.18(d,lH),

5.25(br.s,2H), 2.44(s,3H)

CDC1₃ 7.52(d,lH), 7.32(dd,lH), 7.21(d,lH),

5.08(br.s,2H), 4.66(br.s,2H), 2.42(s,3H)

DMSO-dg 1.9(s,3H,6-CH₃), 2.2(s,3H,N-Me),

2.25-2.40(m,4H,-N N-), 3.55-3.75 (m,4H,-N N-), 5.85(2H₍br.s,-NH₂), 7.2(d,2H,J1.5Hz,2',6'-H), 7.52(dd,1H, J1.5Hz,4'-H)

OMSO-dg 7.58(dd,lH), 7.45(dd,lH), 7.35(d,lH),

7.24(br.s,lH), 3.35(br.s,2H), 3.96(br.s,2H)

DMSO-dg 1.8(s,3H), 5.8(s,2H), 5.95(s,2H),

7.53(s,lH), 7.92(s,lH)

APO 0016 4

DMSO-dg

7.78(d,lH), 7.59(s,lH), 7.36(d,lH),

6.60-6- 47(br.t,1H), 6.25-6.03(br.s,2H), 3.25(q,2H), l.L(t,3H)

In the foregoing, the signals have been abbreviated as follows: s = singlet; d = doublet; dd = doublet of doublets; t = triplet; q = quadruplet; m = multiplet; br.s = broad singlet; br.t = broad triplet.

MJWD/MS/PA1024/22nd November 1989

- 75 Pharmacological Activity

Inhibition of Glutamate release and Inhibition of Rat Liver DHFR

Compounds of Formula (I) were tested for their effect on veratrineevoked release of glutamate from rat brain slices according to the protocol described in Epilepsia 27(5): 490-497, 1986. The protocol for testing for inhibition of DHFR activity was a modification of that set out in Biochemical Pharmacology Vol.20 pp 561-574, 1971.

The results are given in Table 1, the ICgg being the concentration of compound to cause _ 50% inhibition of (a) veratrine-evoked release of glutamate and (b) of DHFR enzyme activity.

TABLE 1

Compound of Example No.	Glutamate Release (P95 Holts)	IC50<^) Rat Liver DHFR (P95 Helts)
1	1.18 (0.50-2.60)	>100
2	0.56 (0.23-1.37)	33 (27.00-40.00)
3	2.15 (0.90-5.10)	>100
4	0.33 (.0.196-0.566)	>30<100
5	3.50 (1.10-10.40)	ca.IQO
6	0.70 (0.40-1.50)	0.51 (0.36-0.73)
7	<10.00	>10.0
8	<10.00	>10.0
9	<10.00	>100.00
10	<10.00	>100.00
11	4.80 (2.30-10.20)	>100.00
12	<10.00	>100.00
13	<10	>100.00

MJWD/MS/PA1024/22nd November 1989

14	3.1 (2.1-4.6)	>100.00
15	2.7 (1.0-7.2)	8.7 (5.2-14.7)
16	3.2 (1.7-6.1)	>100
17	2.4 (1.00-5.80)	4.9 (3.90-6.20)
18	<10.00	ca.100
19	2.6 (0.80-8.50)	>100.00
20	4.2 (1.20-15.30)	17.50(9.80-31.40)
21	11.5 (4.80-27.60)	16.01(12.05-21.282)
22	2.80 (0.80-9.80)	23.800(9.00-61.00)
23	8.70 (2.60-29.10)	20.940(9.00-61.00)
24	2.10 (0.90-4.80)	15.10(11.00-20.70)
25	4.10 (1.10-15.50)	>100.00
26	ca.3.00	clO.OO
27	ca.10.00	>100.00
28	4.6(1.60-13.30)	46.10(14.30-148.90)
29	1.57(0.94-2.62)	0.53(0.348-0.812)

APO00164

MJWD/MS/PA1024/22nd November 1989

- 78 Toxicological Exawl·

The compound of Example 1 has been administered intravenously to groups of six male and six female Wistar rats once daily at dose levels of upto 15mg/kg/day. The no observsed effect dose was 2.5mg/kg/day.

The compound of Example 2, has been rats the no observed effect dose was observed effect dose was 14mg/kg/day.	tested in both rats and dogs.
2.5mg/kg/day a	ind in dogs, the
PharaacMitlcal Forwlatlon Exawl·
Tablet:
INGREDIENT
A: Compound of Example 1	150 mg )
Lactose	200 mg )
Maize Starch	50 mg )
Polyvinyl pyrrolidone	4 mg )
Magnesium Stearate	4 mg )
) = contents per tablet.		•
The C'-ug was mixed with the (actcs-	at stance and	granulated wt·
s:\:;on :* the po 1 yv '..ny Ipyrro ' ;co.· a	;.r «iter. Tne	resultant gran:
MJWD/MS/PA1024/22nd November 1989

·*

BAD ORIGINAL

- 79 were dried, mixed with magnesium stearate and compressed to give tablets.

B: INJECTION (I)

The salt of the compound of Formula I was dissolved in sterile water for injection.

INTRAVENOUS INJECTION FORMULATION (II)

Active ingredient 0.20g

Sterile, pyrogen-free phosphate buffer (pH9.0) to 10ml

The compound of Example I as a salt is dissolved in most of the phosphate buffer at 35-40°C, then made up to volume and filtered through a sterile micropore filter into sterile 10ml glass vials (Type 1) which are sealed with sterile closures and overseals.

In the following Examples, the active compound maybe any compound of formula (I) or pharmaceutically acceptable salt thereof.

C: Capsule formulations

APO00164

Capsule Formulation A

Formulation A may be prepared by admixing the ingredients and filling two-part hard gelatin napsu’es with the resulting mixture.

MJWD/MS/PA1024/22nd November 1989

BAD ORIGINAL

- 80 mq/capsule

(a)	Active ingredient	250
(b)	Lactose B.P.	143
(c)	Sodium Starch Glycol late	25
(d)	Magnesium Stearate	2 420

Capsule Formulation B mq/capsule (a) Active ingredient 250 (b) Macrogel 4000 BP 350

600

Capsules may be prepared by melting the Macrogel 4000 BP, dispersing the active ingredient in the melt, and filling two-part hard gelatin capsules therewith.

Capsule Formulation B (Controlled release capsule) mq/capsule

(a)	Active ingredient	250
(b)	Microcrystalline Cellulose	125
(c)	Lactose BP	125
(d)	Ethyl Cellulose	13
		513

The contro1 led-release capsule formulation may be prepared by extruding mixed ingredients (a) to (c) using an extruder, tne.n spheronising and

MJWD/MS/PA1024/22nd November 1989 —

BAD ORIGINAL drying the extrudate. The dried pellets are coated with ethyl cellulose (d) as a control led-release membrane and filled into two-part hard gelatin capsules.

Syrup formulation

Active ingredient 0.2500 g

Sorbitol Solution 1.5000 g

Glycerol 1.0000 g

Sodium Benzoate 0.0050 g

Flavour 0.0125 ml

Purified Water q.s. to 5.0 ml

The sodium benzoate is dissolved in a portion of the purified water and the sorbitol solution added. The active ' ingredient is aCded and dissolved. The resulting solution is mixed with the glycerol and then made up to the required volume with the purified water.

Suppository formulation mq/suppository

Active ingredient (63μπι)* 250

Hard Fat, BP (Witepsol H15 - Dynamit Nobel) 1770

2020 * The active ingredient is used as a powder wherein at least 90% of the particles are of 53μίτι diameter or less.

MJWD/MS/PA1024/22nd November 1989

- 82 One-fifth of the Witepsol H15 is melted in a steam-jacketed pan at 45°C maximum. The active ingredient is sifted through a 200μπι sieve and added to the molten base with mixing, using a Silverson fitted with a cutting head, until a smooth dispersion is achieved. Maintaining the mixture at 45°C, the remaining Witepsol H15 is added to the suspension which is stirred to ensure a homogenous mix. The entire suspension is then passed through a 250um stainless steel screen and, with continuous stirring, allowed to cool to 40°C. At a temperature of 38-40°C, 2.02g aliquots of the mixture are filled into suitable plastic moulds and the suppositories allowed to cool to room temperature.

MJWD/MS/PA1024/22nd November 1989

Claims (8)

1. Use of a compound of Formula I or a/facid addition salt thereof in the

manufacture of a medicament for or diseases of a mammal, wherein treating or preventing CNS disorders !“* /? p \ Λ5 i\>— Λ λ ^N'· Ή < _zx~- . \ / \ -n 1I / \ \ / t \ \ \ h/ λ, *7

where R hydrogen^ to which and in Formula I,

R| and are the same or different and are selected from halo, hydroxy, alkoxy, *lhy >, alkylthio and a group NR^R^ and rH are the same or different and are selected from alkyl, aryn-iand >eyla1i>yl or together with the nitrogen atom group from M-al k^l pi'peraxino they are attached form a ayalia ring opfriana; ly^uhif if'ifod with-aaa. ¹ morphol«no and piparictin© · =aa-mors alkyl, greupa and-apt rone Vly .aanta-ifting-a-.-ifurthe'* he-fr-rnnaea;

Rg is hydrogen, alkyl optionally substituted by one or more halo

SelecVed fnorc radicals, or is ) alliywMKwe, '44a 1 Itylawiw», cyano, halo, carbamoyl, hydroxy, ea»bo*yj- ' alkoxy, alkylthio, alkylthioalkyl,

Η R di(alkyloxy)alkyl, -C(A):NOH ex -CCJ^ er -CCLR wherein R is and.

hydrogen or alkyl, [sr a group CF^X where X is hydroxy, alkoxy, phe.doxy benzwloxu °^r i ii i n ®-^acb ^a*kw( · .anylfitxy, aojtlaTly 1 aiQr, Μ«, cyano^ -NR R wherein R and R are >

MJW0/MS/PA1024-EP/22nd November 1989

2.S A precess for the production of a compound or an acidjsalt thereof according to any one of claims 9 to 4)^ which comprises the reaction a compound of formula II

II c

c c

c <

L.

cchorn or wherein Rg to Rg are as hereinbefore defined, L is a leaving ^group, and Y is cyanojar^carboxy, cirhnnyl, or alkoxycarbonyl with a compound of formula III or So.IV Hiere©^· κ H \\

-C-K

III £ollow«jd as appropriate bj Gonuersian Hie pre dux V wherein R^ is as hereindef ined, and ^aelating-tha e£mp8und--ot· Fj|rffiulaInbo the free base or an acid addition salt thereof,-and ;mp»i

-ueoftvegiing tha bm intO'tn-tcidadd-ition xlt-tharaaf ar Tntc irw^s

MJWD/MS/PA1024-EP/22nd November 1989

BAD ORIGINAL

2,4-Diamino-5-(2,3,5-trichloropheny1)pyrimidine y^ and

23 4-Amino-5-(3,5-dichlorophenyl)-6-methyl-2-(4-methylpiperazin-l· y 1 )pyrimidine

2.4- 0iamino-5-(2,3-dichlorophenyl)-6-methy1pyrimidine and ai. 2 , 4-0 i ami no-5-(2,3-dichlorophenyl )-6-methoxymethylpyr imidine 2,4-0iamino-5-(2,4-dich loropheny1)-6-trif luoromethylpyrimidine δ-3enzylox vmethyl-2,4-diami no-5-(2,4-dich loropheny1) pyrimidine

Cm- ' -iand

2X. 2-^-methy 1 piperazir^ j^4-amino-5-(2,4-d ich lorophenyl)pyrimidine 2,4-0iamino-5-(2,5-dich lorophenyl)-6-trif luoromethylpyrimidine

2.4- 0 i ami no-5-(2,3-d ich lorophenyl )-6-trif luoromethyl pyrimidine and ·.

2,4-0iami no-5-(2,3,5-trichloropheny!)-6-methyl pyrimidine

?2 '^*^ils?l * i<<'Λ··Rg is hydrogen or halo.

'SO_aNfc'ft‘'

-aauae, or -group

AP 0 0 0 1 6 4 in claim 1

13. A compound or acid addition salt/of formula I/wherein Rp R₂, R., Rg, , , p*e/\ox e^oxy *=e*?,

Rg, Ry and Rg are as defined in claim 1 and R^ is/alkoxy, banxyloxw f- --;-ι iiry1i Tltylarfy or alkylthio. I a <^roof CH_aX uiKene. X <s \

14. 4-Amino-2-(4-methylpiperazin-l-yl)-5-(2,3,5-trichioropheny!)-6p^rimidinc and s trif lucromethyl acid addition salt^ bkerec^..

and acid addition salt.

H\eceof .

MJWD/MS/PA1024-EP/22nd November 1989

15. 2,4-Diami no-5-(2,3,5-trichlorophenyl)-6-methoxymethylpyrimidine £

2.

accorating ha c(a‘«nh 1 of o Cor^pownd or So.Ik of

Use oi-t-ceapounri^Hi f-iatflied ia claim 1 wherein one of R^ and R₂ is amino, and the other is selected from amino, hydroxy, halo, o

morpholino, pi pern riajtU N-alkylpiperazinjfi, N,N-dialkylamino, ond

Ν,-alkylamino alkylthio;

Rg is alkyl optionally substituted by one or more halo radicals, or is ^Λ '^7-, alkylthio, hydrogen, hydroxy, ja.lJrjOJjt, halo, carboxy, carbamoyl, or a group C^X where X is hydroxy, phenoxy, benzyloxy, alkoxy or alkylthio; _ancj one of R₄ and Rg is halo and the other is selected from halo hydrogen;

or

I¹ λ It

Rg is halo, hydrogen^ nitro nr naiaa·; SO_aNR'ft'

R. is hydrogen, halo, cyano, alkylthio, .WgiUS-k* iilky.l, nYee-, ^el-kalcu{phon«4| amino or woth.->nyn1phnwnmi«1n; ana

MJW0/MS/PA1024-EP/22nd November 1989

-

- 2 PROOUCT/EP CLAIMS

Mgr ined ab'.

i no >*0i.

tune·· each of R₄ to Rg

-tha.

tamp f!n—

UUXj iat if.dffesi, cyano, hydrogen, halo, atk^lMvo <ind (, a 1 ky T su I phony 1 ami no, ^=iae is selected from o w y, uR >

(gSfcj

T7' or ^J · · · n | SO^N^¹¹ cokers. fc'ond fc cure eack

R₄ and Rg or R. and Rg together are the group -C'H=CH-CH=CHijaragji^» in which case both R- and Rg are hydrogen;

and optionally one of the nitrogen atoms in the pyrimidine ringa»s

Hve

N alkylated or w-t^oriB-riyta? ae^iri N oxide;

ΟλΗ atky?

the foregoing alkyl groups F moieties of alkyl-containing groups iS having from 1 to J^carbon atoms»-*aa the aryl graupa of ary» cantaining··group»· naming ό

3 PROOUCT/EP CLAIMS

Rg is hydrogen or halo.

according Vo claim i of a compound or sail· of3. Use mp—a compound ffkimad in claims 1 or 2 wherein R. is hydroxy, or amino, N-alkylamino, Ν,Ν-dialkylamino, morpholino, -ftipor^-nnyl, N-alkylpiperazin^ .

according Vo claim 1 of a compound or sail· of·

4-Amino-2-J-methylaminc-5-(2,3,5-trichlorophenyl) pyrimidine and $ ar^wa acid addition salthereof.

f or q, pKctrmo.ct.u.ViCall^ acceptable acid add i hop sail· au X A pharmaceutical formulation I comprising a compound according to any one ο* claims 9 to ή^/together with one or more acceptable carriers therefor and optionally other therapeutic ingredients.

addih' on

4-Amino-2-/^ethy lamino)-5-(2,3,5-tr ich lorophenyl )pyr imidine MJWD/MS/PA1024-EP/22nd November 1989

BAD ORIGINAL

4-Aminc-2-N.-mcroho I i no-5-(2,3,5-tr ich loropheny! )-6-tri fluoromethyl pyrimidine and acid addif-ion Sad'S K.ere.o£.

iq 4-Ami no-Z-N^-dimethyl amino-5-(2,3,5-tr ich loropheny 1)-6trif luoromethylpyrimidine

- M4-Amino-2 jTmorpnol ino-^-5-(2,3,5-tr ich lorophenyl) pyrimidine 4-Ami no-2-N,N-d imethyl amino-5-(2,3,5-tr ich loropheny 1) pyrimidine \

2o. 4-Amino-6-methyl-2-(4-methylpiperazin-l-y1)-5-(2,3,5-tr ich 1 orcphenyl )pyrimidine

- 4 PROOUCT/EP CLAIMS

Rg is hydrogen or R^ and Rg are both hydrogen, Rg and R? are both chloro and Rg is selected from hydrogen, chloro and nitro.

6p

8. Use of a compound af-fnrm» la I or ** .•wMiKnw salt th*£e^f=as ^dei±aed=ia claim 1 in the manufacture of a medicament for treating or preventing CNS disorders or diseases of a mammal wherein R^ to Rg are as defined in claim 1 with the proviso that when Ry is halo, then

Rg is hydrogen, perhaloalkyl, methyl or methoxymethyl and/or

R_c is nitro and/or ⁶ O

R| is N.-alkylpiperazinj^ , morpholino, N,^-dimethylamino, pipppnaiajtl· or N.-ethylamino; or with the proviso that when Rg is chloro then

R^ is halo, and Rg is hydrogen, perhaloalkyl, methoxymethyl, methyl or o

halo and/or Rj is N-methylpiperazin^B, morphol ino^er N..Ndimethylamino, or N-ethylamino.

9. A compound or acid addition salt thereof of Formula I as=^e£±eed in claim 1 with the proviso that at least one of R^ to Rg is other than hydrogen, and further with the proviso that when R^ and R₂ are both amino, or when Rj, is hydroxy and R^ is amino, and Rg is alkyl or hydrogen, and R? is hydrogen, then R^ and Rg are both halo.

MJW0/MS/PA1024-EP/22nd November 1989

4. Use of--n ccmpftttftrf a^.rllimed in any one of claims 1 to 3 wherein R₂ is chloro, amino, N,N-dia Iky1 ami no, or piperidino.

according Vo claim 1 of a Compound or SalV of

- 5 PRCOUCT/E? CLAIMS in claim 1

10. A comocund or acid addition salt thereof of Formula I^wherein R^ to R, and one o*r and is ©Pkcr Fka.n ivjdrcgen.

and R_a are as defined in claim l^and R₇ is halo, «fed cyanc, t, a Iky 1th io, j^·

IaH or so.nk'R

4r^- X hhcreof· ·π claim i.

11. A ccmccund or acid addition saltjfof formula I^wherein R^ to Rg are as defined in claim 1 and R. is morpholine, a i aawaa iw;·^ O ¹ ec

N-a 1 ky 1 piperazin^ , Ν,Ν-dia ikylamino^ N.-a 1 ky 1 amino_t't-j 1 h^;nH\e.re.o*r ^,n claim i

12. A compound or acid addition salt/of formula i/wherein R. and R? are as

Se.lec.fed from defined in claim 1 and R, is h-lkeie», alkylthio sr alkyl substituted by ^J * and ^J one or more halo radicals, ta* a group CHgX where X is alkylthio, phenox^j benzol oxy jjg.ysayy, ftp.yΙ.ΐ.i¥y, alkyloxy or hydroxy;

R. to R ,.frhr> ,.-.,-tsin .m 4».t-7i«noi .-md are each selected from ^{4 6} and hydrogen, halo, Nry¹, ~yn. nitro;

Ry is ha 1 o, adkyl, per h a1sa-lfatg, cyano, s »'}! 11 \ , ι _-111_:

5. Use of··· compound ^t-claiin»d=« any one of claims 1 to 4 wherein Rg is hydrogen, alkyl, methoxymethyl, trifluoromethyl, benzyloxymethyl, phenoxymethyl or methylthiomethyl.

according Vo claim 1 of a compound or Sail· of

- 6 PROOUCT/EP CLAIMS

Ιό. A-Amino-2-(4-methylpiperazin-l-yl)-5-(2,3,5-trichlorophenyl an4 s pyrimidine «£· acid addition salt^ B\ers.C|-.

αη4

17. 2,4-0iami no-5-(2,3,5-trichlorophenyl )-6-tri fluoromethyl pyrimidine £ acid addition salt^ i4\<rc.o£.

18. 2,4-Qiamino-5-(4-nitro-2,3,5-trichlorophenyl) pyrimidine

6. Use afr-jfr..,ccapi3und ai-xlairrud in claims 1 or 2 wherein one of R. and R_o ¹ °^Z is amino and the other is amino, piperneiftyl or N.-methylpiperazinrf, or

N.-alkylamino£ Ν,,Ν-dialkylamino and

Rg is hydrogen, methyl, trifluoromethyl or methoxymethyl.

according Vo claim 1 of A Compound or SalV of

- 7 PROOUCT/E? CLAIMS

7. Use of. a compound—ai> elnimcd in claim 1 wherein R. is selected from

O ¹ amino, -.pi proa tiny!, N-methylpiperazin^, ^morpholino,

Ν,Ν-dimethylamino, and N-ethylamino;

cP <P

CP

CP oR₂ is amino;

Rg is selected from trifluoromethyl, hydrogen, methyl, benzyloxymethyl, methoxymethyl and methylthiomethyl;

R^ is chloro; and at least one of Rg, Rg and R? is chloro, and the remainder are selected from hydrogen, chloro and nitro; and

MJWD/MS/PA1024-EP/22nd November 1989

- 8 PRODUCT/EP CLAIMS nimnwH ι» Γ·ιι~»η.la. f·.. <»« »» »^.w«*1» »np^oi..

O£_ A compound or pharmaceutically acceptable / salt thereof as claimed in • S 23 ^K acid ad dip ion icceptable £sa1t claim^9 to for use in tn«rapy. medicine..