The invention concerns quinazoline derivatives of formula wherein n is 1, 2 or 3 and each R.sup.2 is independently halogeno; and R.sup.1 is di-(1-4C)alkyl!amino-(2-4C)alkoxy, pryolidin-1-yl-(2-4C)alkoxy,piperidino-(2-4C)alkoxy, morpholino-(2-4C)alkoxy, piperazin-1-yl-(2-4C)alkoxy or 4-(1-4C)alkylpiperazin-1-yl-(2-4C)alkoxy; or pharmaceutically-acceptable salts thereof; processes for their preparation, pharmaceutical compositions containing them, and the use of the receptor tyrosine kinase inhibitory properties of the compounds in the treatment of proliferative disease such as cancer.

The invention relates to quinazoline derivatives, or 
pharmaceutically-acceptable salts thereof, which possess 
anti-proliferative activity such as anti-cancer activity and are 
accordingly useful in methods of treatment of the human or animal body. 
The invention also relates to processes for the manufacture of said 
quinazoline derivatives, to pharmaceutical compositions containing them 
and to their use in the manufacture of medicaments of use in the 
production of an anti-proliferative effect in a warm-blooded animal such 
as man. 
Many of the current treatment regimes for cell proliferation diseases such 
as psoriasis and cancer utilise compounds which inhibit DNA synthesis. 
Such compounds are toxic to cells generally but their toxic effect on 
rapidly dividing cells such as tumour cells can be beneficial. Alternative 
approaches to anti-proliferative agents which act by mechanisms other than 
the inhibition of DNA synthesis have the potential to display enhanced 
selectivity of action. 
In recent years it has been discovered that a cell may become cancerous by 
virtue of the transformation of a portion of its DNA into an oncogene i.e. 
a gene which, on activation, leads to the formation of malignant tumour 
cells (Bradshaw, Mutagenesis, 1986, 1, 91). Several such oncogenes give 
rise to the production of peptides which are receptors for growth factors. 
The growth factor receptor complex subsequently leads to an increase in 
cell proliferation. It is known, for example, that several oncogenes 
encode tyrosine kinase enzymes and that certain growth factor receptors 
are also tyrosine kinase enzymes (Yarden et al., Ann. Rev. Biochem., 1988, 
57, 443; Larsen al. Ann. Reports in Med. Chem. 1989, Chpt. 13). 
Receptor tyrosine kinases are important in the transmission of biochemical 
signals which initiate cell replication. They are large enzymes which span 
the cell membrane and possess an extracellular binding domain for growth 
factors such as epidermal growth factor (EGF) and an intracellular portion 
which functions as a kinase to phosphorylate tyrosine amino acids in 
proteins and hence to influence cell proliferation. Various classes of 
receptor tyrosine kinases are known (Wilks, Advances in Cancer Research, 
1993, 60, 43-73) based on families of growth factors which bind to 
different receptor tyrosine kinases. The classification includes Class I 
receptor tyrosine kinases comprising the EGF family of receptor tyrosine 
kinases such as the EGF, TGF.alpha., NEU, erbB, Xmrk, HER and let23 
receptors, Class II receptor tyrosine kinases comprising the insulin 
family of receptor tyrosine kinases such as the insulin, IGFI and 
insulin-related receptor (IRR) receptors and Class III receptor tyrosine 
kinases comprising the platelet-derived growth factor (PDGF) family of 
receptor tyrosine kinases such as the PDGF.alpha., PDGF.beta. and 
colony-stimulating factor 1 (CDF1) receptors. It is known that Class I 
kinases such as the EGF family of receptor tyrosine kinases are frequently 
present in common human cancers such as breast cancer (Sainsbury et al., 
Brit J. Cancer, 1988, 58, 458; Guerin et al., Oncogene Res., 1988, 3, 21 
and Klijn et al., Breast Cancer Res. Treat., 1994, 29, 73), non-small cell 
lung cancers (NSCLCs) including adenocarcinomas (Cerny et al., Brit. J. 
Cancer, 1986, 54, 265; Reubi et al., Int. J. Cancer, 1990, 45, 269; and 
Rusch et al., Cancer Research, 1993, 53, 2379) and squamous cell cancer of 
the lung (Hendler et al., Cancer Cells, 1989, 7, 347), bladder cancer 
(Neal et al., Lancet, 1985, 366), oesophageal cancer (Mukaida et al., 
Cancer, 1991, 68, 142), gastrointestinal cancer such as colon, rectal or 
stomach cancer (Bolen et al., Oncogene Res., 1987, 1, 149), cancer of the 
prostate (Visakorpi et al., Histochem. J., 1992, 24, 481), leukaemia 
(Konaka et al., Cell, 1984, 37, 1035) and ovarian, bronchial or pancreatic 
cancer (European Patent Specification No. 0400586). As further human 
tumour tissues are tested for the EGF family of receptor tyrosine kinases 
it is expected that their widespread prevalance will be established in 
further cancers such as thyroid and uterine cancer. It is also known that 
EGF type tyrosine kinase activity is rarely detected in normal cells 
whereas it is more frequently detectable in malignant cells (Hunter, Cell, 
1987, 50, 823). It has been shown more recently (W J Gullick, Brit. Med. 
Bull., 1991, 47, 87) that EGF receptors which possess tyrosine kinase 
activity are overexpressed in many human cancers such as brain, lung 
squamous cell, bladder, gastric, breast, head and neck, oesophageal, 
gynaecological and thyroid tumours. 
Accordingly it has been recognised that an inhibitor of receptor tyrosine 
kinases should be of value as a selective inhibitor of the growth of 
mammalian cancer cells (Yaish et al. Science, 1988, 242, 933). Support for 
this view is provided by the demonstration that erbstatin, an EGF receptor 
tyrosine kinase inhibitor, specifically attenuates the growth in athymic 
nude mice of a transplanted human mammary carcinoma which expresses EGF 
receptor tyrosine kinase but is without effect on the growth of another 
carcinoma which does not express EGF receptor tyrosine kinase (Toi et al., 
Eur. J. Cancer Clin. Oncol., 1990, 26, 722.) Various derivatives of 
styrene are also stated to possess tyrosine kinase inhibitory properties 
(European Patent Application Nos. 0211363, 0304493 and 0322738) and to be 
of use as anti-tumour agents. The in vivo inhibitory effect of two such 
styrene derivatives which are EGF receptor tyrosine kinase inhibitors has 
been demonstrated against the growth of human squamous cell carcinoma 
inoculated into nude mice (Yoneda et al., Cancer Research, 1991, 51, 
4430). Various known tyrosine kinase inhibitors are disclosed in a more 
recent review by T R Burke Jr. (Drugs of the Future, 1992, 17, 119). 
It is known from European Patent Applications Nos. 0520722, 0566226 and 
0635498 that certain quinazoline derivatives which bear an anilino 
substituent at the 4-position possess receptor tyrosine kinase inhibitory 
activity. It is further known from European Patent Application No. 0602851 
that certain quinazoline derivatives which bear a heteroarylamino 
substituent at the 4-position also possess receptor tyrosine kinase 
inhibitory activity. 
It is further known from International Patent Application WO 92/20642 that 
certain aryl and heteroaryl compounds inhibit EGF and/or PDGF receptor 
tyrosine kinase. There is the disclosure of certain quinazoline 
derivatives therein but no mention is made of 4-anilinoquinazoline 
derivatives. 
The in vitro anti-proliferative effect of a 4-anilinoquinazoline derivative 
has been disclosed by Fry et al., Science, 1994, 265, 1093. It was stated 
that the compound 4-(3'-bromoanilino)-6,7-dimethoxyquinazoline was a 
highly potent inhibitor of EGF receptor tyrosine kinase. 
The in vivo inhibitory effect of a 4,5-dianilinophthalimide derivative 
which is an inhibitor of the EGF family of receptor tyrosine kinases has 
been demonstrated against the growth in BALB/c nude mice of a human 
epidermoid carcinoma A-431 or of a human ovarian carcinoma SKOV-3 
(Buchdunger et al., Proc. Nat. Acad. Sci., 1994, 91, 2334). 
It is further known from European Patent Application No. 0635507 that 
certain tricyclic compounds which comprise a 5- or 6-membered ring fused 
to the benzo-ring of a quinazoline possess receptor tyrosine kinase 
inhibitory activity. It is also known from European Patent Application No. 
0635498 that certain quinazoline derivatives which carry an amino group at 
the 6-position and a halogeno group at the 7-position possess receptor 
tyrosine kinase inhibitory activity. 
Accordingly it has been indicated that Class I receptor tyrosine kinase 
inhibitors will prove to be useful in the treatment of a variety of human 
cancers. 
It is also expected that inhibitors of EGF type receptor tyrosine kinases 
will be useful in the treatment of other diseases of excessive cellular 
proliferation such as psoriasis (where TGF.alpha. is believed to be the 
most important growth factor) and benign prostatic hypertrophy (BPH). 
There is no disclosure in these documents of quinazoline derivatives which 
bear at the 4-position a halogeno-substituted anilino substituent and 
which also bear a dialkylaminoalkoxy substituent at the 6-position. We 
have now found that such compounds possess potent in vivo 
anti-proliferative properties which are believed to arise from their Class 
I receptor tyrosine kinase inhibitory activity. 
According to the present invention there is provided a quinazoline 
derivative of the formula I 
##STR1## 
wherein n is 1, 2 or 3 and each R.sup.2 is independently halogeno; and 
R.sup.1 is di-(1-4C)alkyl!amino-(2-4C)alkoxy, 
pyrrolidin-1-yl-(2-4C)alkoxy, piperidino-(2-4C)alkoxy, 
morpholino-(2-4C)alkoxy, piperazin-1-yl-(2-4C)alkoxy or 
4-(1-4C)alkylpiperazin-1-yl-(2-4C)alkoxy; 
or a pharmaceutically-acceptable salt thereof. 
In this specification the term "alkyl" includes both straight and branched 
chain alkyl groups but references to individual alkyl groups such as 
"propyl" are specific for the straight chain version only. For example 
when R.sup.1 is a di-(1-4C)alkyl!amino-(2-4C)alkoxy group, suitable 
values for this generic radical include 2-dimethylaminoethoxy, 
3-dimethylaminopropoxy, 2-dimethylaminopropoxy and 
1-dimethylaminoprop-2-yloxy. An analogous convention applies to other 
generic terms. 
Within the present invention it is to be understood that, insofar as 
certain of the compounds of the formula I may exist in optically active or 
racemic forms by virtue of one or more substituents containing an 
asymmetric carbon atom, the invention encompasses any such optically 
active or racemic form which possesses anti-proliferative activity. The 
synthesis of optically active forms may be carried out by standard 
techniques of organic chemistry well known in the art, for example by 
synthesis from optically active starting materials or by resolution of a 
racemic form. 
The quinazolines of the formula I are unsubstituted at the 2-, 5- and 
8-positions. 
It is also to be understood that certain quinazoline derivatives of the 
formula I can exist in solvated as well as unsolvated forms such as, for 
example, hydrated forms. It is to be understood that the invention 
encompasses all such solvated forms which possess anti-proliferative 
activity. 
Suitable values for the generic radicals referred to above include those 
set out below. 
A suitable value for R.sup.2 when it is halogeno is, for example fluoro, 
chloro, bromo or iodo. 
Suitable values for each R.sup.1 substituent which may be present on the 
quinazoline ring include, for example: 
______________________________________ 
for di-(1-4C)alkyl!amino- 
2-dimethylaminoethoxy, 
(2-4C)alkoxy: 2-(N-ethyl-N-methylamino)ethoxy, 
2-diethylaminoethoxy, 
2-dipropylaminoethoxy, 
3-dimethylaminopropoxy, 
3-diethylaminopropoxy, 
2-dimethylaminopropoxy, 
2-diethylaminopropoxy, 
1-dimethylaminoprop-2-yloxy, 
1-diethylaminoprop-2-yloxy, 
1-dimethylamino-2-methylprop-2-yloxy, 
2-dimethylamino-2-methylpropoxy, 
4-dimethylaminobutoxy, 
4-diethylaminobutoxy, 
3-dimethylaminobutoxy, 
3-diethylaminobutoxy, 
2-dimethylaminobutoxy, 
2-diethylaminobutoxy, 
1-dimethylaminobut-2-yloxy and 
1-diethylaminobut-2-yloxy; 
for pyrrolidin-1-yl-(2-4C)- 
2-(pyrrolidin-1-yl)ethoxy, 
alkoxy: 3-(pyrrolidin-1-yl)propoxy and 
4-(pyrrolidin-1-yl)butoxy; 
for piperidino-(2-4C)alkoxy: 
2-piperidinoethoxy, 
3-piperidinopropoxy and 
4-piperidinobutoxy; 
for morpholino-(2-4C)alkoxy: 
2-morpholinoethoxy, 
3-morpholinopropoxy and 
4-morpholinobutoxy; 
for piperazin-1-yl-(2-4C)alkoxy: 
2-(piperazin-1-yl)ethoxy, 
3-(piperazin-1-yl)propoxy and 
4-(piperazin-1-yl)butoxy; 
for 4-(1-4C)alkylpiperazin-1-yl- 
2-(4-methylpiperazin-1-yl)ethoxy, 
(2-4C)alkoxy: 3-(4-methylpiperazin-1-yl)propoxy and 
4-(4-methylpiperazin-1-yl)butoxy. 
______________________________________ 
A suitable pharmaceutically-acceptable salt of a quinazoline derivative of 
the invention is, for example, a mono- or di-acid-addition salt of a 
quinazoline derivative of the invention which is sufficiently basic, for 
example, an acid-addition salt with, for example, an inorganic or organic 
acid, for example hydrochloric, hydrobromic, sulphuric, phosphoric, 
trifluoroacetic, citric, maleic, tartaric, fumaric, methanesulphonic or 
4-toluenesulphonic acid. 
Particular novel compounds of the invention include, for example, 
quinazoline derivatives of the formula I, or pharmaceutically-acceptable 
salts thereof, wherein: 
(a) n is 1 or 2 and each R.sup.2 is independently fluoro, chloro or bromo; 
and R.sup.1 has any of the meanings defined hereinbefore or in this 
section relating to particular novel compounds of the invention; 
(b) R.sup.1 is 2-dimethylaminoethoxy, 2-diethylaminoethoxy, 
3-dimethylaminopropoxy, 3-diethylaminopropoxy, 2-(pyrrolidin-1-yl)ethoxy, 
3-(pyrrolidin-1-yl)propoxy, 2-piperidinoethoxy, 3-piperidinopropoxy, 
2-morpholinoethoxy, 3-morpholinopropoxy, 2-(piperazin-1-yl)ethoxy, 
3-(piperazin-1-yl)propoxy, 2-(4-methylpiperazin-1-yl)ethoxy or 
3-(4-methylpiperazin-1-yl)propoxy; 
and n and R.sup.2 have any of the meanings defined hereinbefore or in this 
section relating to particular novel compounds of the invention. 
A preferred compound of the invention is a quinazoline derivative of the 
formula I 
wherein (R.sup.2).sub.n is 3'-chloro, 3'-bromo, 2',4'-difluoro, 
2',4'-dichloro, 3',4'-difluoro, 3',4'-dichloro, 3'-fluoro-4'-chloro or 
3'-chloro-4'-fluoro; 
and R.sup.1 is 2-dimethylaminoethoxy, 2-diethylaminoethoxy, 
3-dimethylaminopropoxy, 3-diethylaminopropoxy, 2-(pyrrolidin-1-yl)ethoxy, 
3-(pyrrolidin-1-yl)propoxy, 2-piperidinoethoxy, 3-piperidinopropoxy, 
2-morpholinoethoxy or 3-morpholinopropoxy; 
or a pharmaceutically-acceptable acid-addition salt thereof. 
A further preferred compound of the invention is a quinazoline derivative 
of the forula I 
wherein (R.sup.2).sub.n is 3',4'-difluoro or 3'-chloro-4'-fluoro; 
and R.sup.1 is 3-dimethylaminopropoxy, 3-diethylaminopropoxy, 
3-(pyrrolidin-1-yl)propoxy, 3-piperidinopropoxy or 3-morpholinopropoxy; 
or a pharmaceutically-acceptable acid-addition salt thereof. 
According to a further aspect of the present invention there is provided 
the quinazoline derivative 
4-(3'-chloro-4'-fluoroanilino)-6-(3-diethylaminopropoxy)quinazoline; 
or a pharmaceutically-acceptable salt thereof. 
According to a further aspect of the present invention there is provided 
the quinazoline derivative 
4-(3',4'-difluoroanilino)-6-(3-morpholinopropoxy)quinazoline; 
or a pharmaceutically-acceptable salt thereof. 
It is also to be understood that these quinazoline derivatives of the 
invention can exist in solvated as well as unsolvated forms such as, for 
example, hydrated forms. It is to be understood that the invention 
encompasses all such solvated forms which possess anti-proliferative 
activity. 
The quinazoline derivatives of the invention, or 
pharmaceutically-acceptable salts thereof, may be prepared by any process 
known to be applicable to the preparation of chemically-related compounds. 
Suitable processes include, for example, those illustrated in European 
Patent Applications Nos. 0520722, 0566226, 0602851, 0635498 and 0635507. 
Such processes, when used to prepare the quinazoline derivatives of the 
invention, or pharmaceutically-acceptable salts thereof, are provided as a 
further feature of the invention and are illustrated by the following 
representative Examples. Necessary starting materials may be obtained by 
standard procedures of organic chemistry. The preparation of such starting 
materials is described within the accompanying non-limiting Examples. 
Alternatively necessary starting materials are obtainable by analogous 
procedures to those illustrated which are within the ordinary skill of an 
organic chemist. 
(a) The reaction, conveniently in the presence of a suitable base, of a 
quinazoline of the formula II 
##STR2## 
wherein Z is a displaceable group, with an aniline of the formula III 
##STR3## 
A suitable displaceable group Z is, for example, a halogeno, alkoxy, 
aryloxy or sulphonyloxy group, for example a chloro, bromo, methoxy, 
phenoxy, methanesulphonyloxy or toluene-4-sulphonyloxy group. 
A suitable base is, for example, an organic amine base such as, for 
example, pyridine, 2,6-lutidine, collidine, 4-dimethylaminopyridine, 
triethylamine, morpholine, N-methylmorpholine or 
diazabicyclo5.4.0!undec-7-ene, or, for example, an alkali or alkaline 
earth metal carbonate or hydroxide, for example sodium carbonate, 
potassium carbonate, calcium carbonate, sodium hydroxide or potassium 
hydroxide. Alternatively a suitable base is, for example, an alkali metal 
or alkaline earth metal amide, for example sodium amide or sodium 
bis(trimethylsilyl)amide. 
The reaction is preferably carried out in the presence of a suitable inert 
solvent or diluent, for example an alkanol or ester such as methanol, 
ethanol, isopropanol or ethyl acetate, a halogenated solvent such as 
methylene chloride, chloroform or carbon tetrachloride, an ether such as 
tetrahydrofuran or 1,4-dioxan, an aromatic solvent such as toluene, or a 
dipolar aprotic solvent such as N,N-dimethylformamide, 
N,N-dimethylacetamide, N-methylpyrrolidin-2-one or dimethylsulphoxide. The 
reaction is conveniently carried out at a temperature in the range, for 
example, 10 to 150.degree. C., preferably in the range 20 to 80.degree. C. 
The quinazoline derivatives of the invention may be obtained from this 
process in the form of the free base or alternatively they may be obtained 
in the form of a salt with the acid of the formula H-Z wherein Z has the 
meaning defined hereinbefore. When it is desired to obtain the free base 
from the salt, the salt may be treated with a suitable base as defined 
hereinbefore using a conventional procedure. 
(b) For the production of the compounds of the formula I wherein R.sup.1 is 
an amino-substituted (2-4C)alkoxy group, the alkylation, conveniently in 
the presence of a suitable base as defined hereinbefore, of a quinazoline 
derivative of the formula I wherein R.sup.1 is a hydroxy group. 
The reaction is preferably carried out in a suitable inert solvent or 
diluent as defined hereinbefore and at a temperature in the range, for 
example, 0 to 150.degree. C., conveniently at or near 90.degree. C. 
(c) For the production of those compounds of the formula I wherein R.sup.1 
is an amino-substituted (2-4C)alkoxy group, the reaction, conveniently in 
the presence of a suitable base as defined hereinbefore, of a compound of 
the formula I wherein R.sup.1 is a hydroxy-(2-4 C) alkoxy group, or a 
reactive derivative thereof, with an appropriate amine. 
A suitable reactive derivative of a compound of the formula I wherein 
R.sup.1 is a hydroxy-(2-4C)alkoxy group is, for example, a halogeno- or 
sulphonyloxy-(2-4C)alkoxy group such as a bromo- or 
methanesulphonyloxy-(2-4C)alkoxy group. 
The reaction is preferably carried out in the presence of a suitable inert 
solvent or diluent as defined hereinbefore and at a temperature in the 
range, for example, 10 to 150.degree. C., conveniently at or near 
50.degree. C. 
When a pharmaceutically-acceptable salt of a quinazoline derivative of the 
invention is required, for example a mono- or di-acid-addition salt of the 
quinazoline derivative of the invention, it may be obtained, for example, 
by reaction of said compound with, for example, a suitable acid using a 
conventional procedure. 
As stated hereinbefore the quinazoline derivatives defined in the present 
invention possess anti-proliferative activity which is believed to arise 
from the Class I receptor tyrosine-kinase inhibitory activity of the 
compounds. These properties may be assessed, for example, using one or 
more of the procedures set out below: 
(a) An in vitro assay which determines the ability of a test compound to 
inhibit the enzyme EGF receptor tyrosine kinase. Receptor tyrosine kinase 
was obtained in partially purified form from A-431 cells (derived from 
human vulval carcinoma) by the procedures described below which are 
related to those described by Carpenter et al., J. Biol. Chem., 1979, 254, 
4884, Cohen et al., J. Biol. Chem., 1982, 257, 1523 and by Braun et al., 
J. Biol. Chem., 1984, 259, 2051. 
A-431 cells were grown to confluence using Dulbecco's modified Eagle's 
medium (DMEM) containing 5% fetal calf serum (FCS). The obtained cells 
were homogenised in a hypotonic borate/EDTA buffer at pH 10.1. The 
homogenate was centrifuged at 400 g for 10 minutes at 0-4.degree. C. The 
supernatant was centrifuged at 25,000 g for 30 minutes at 0-4.degree. C. 
The pelleted material was suspended in 30 mM Hepes buffer at pH 7.4 
containing 5% glycerol, 4 mM benzamidine and 1% Triton X-100, stirred for 
1 hour at 0-4.degree. C., and recentrifuged at 100,000 g for 1 hour at 
0-4.degree. C. The supernatant, containing solubilised receptor tyrosine 
kinase, was stored in liquid nitrogen. 
For test purposes 40 .mu.l of the enzyme solution so obtained was added to 
a mixture of 400 .mu.l of a mixture of 150 mM Hepes buffer at pH 7.4. 500 
.mu.M sodium orthovanadate, 0.1% Triton X-100, 10% glycerol, 200 .mu.l 
water, 80 .mu.l of 25 mM DTT and 80 .mu.l of a mixture of 12.5 mM 
manganese chloride, 125 mM magnesium chloride and distilled water. There 
was thus obtained the test enzyme solution. 
Each test compound was dissolved in dimethylsulphoxide (DMSO) to give a 50 
mM solution which was diluted with 40 mM Hepes buffer containing 0.1% 
Triton X-100, 10% glycerol and 10% DMSO to give a 500 .mu.M solution. 
Equal volumes of this solution and a solution of epidermal growth factor 
(EGF; 20 .mu.g/ml) were mixed. 
.gamma.-.sup.32 P!ATP (3000 Ci/mM, 250 .mu.Ci) was diluted to a volume of 
2 ml by the addition of a solution of ATP (100 .mu.M) in distilled water. 
An equal volume of a 4 mg/ml solution of the peptide 
Arg-Arg-Leu-Ile-Glu-Asp-Ala-Glu-Tyr-Ala-Ala-Arg-Gly in a mixture of 40 mM 
Hepes buffer at pH 7.4, 0.1% Triton X-100 and 10% glycerol was added. 
The test compound/EGF mixture solution (5 .mu.l) was added to the test 
enzyme solution (10 .mu.l) and the mixture was incubated at 0-4.degree. C. 
for 30 minutes. The ATP/peptide mixture (10 .mu.l) was added and the 
mixture was incubated at 25.degree. C. for 10 minutes. The phosphorylation 
reaction was terminated by the addition of 5% trichloroacetic acid (40 
.mu.l) and bovine serum albumin (BSA; 1 mg/ml, 5 .mu.l). The mixture was 
allowed to stand at 4.degree. C. for 30 minutes and then centrifuged. An 
aliquot (40 .mu.l) of the supernatant was placed onto a strip of Whatman p 
81 phosphocellulose paper. The strip was washed in 75 mM phosphoric acid 
(4.times.10 ml) and blotted dry. Radioactivity present in the filter paper 
was measured using a liquid scintillation counter (Sequence A). The 
reaction sequence was repeated in the absence of the EGF (Sequence B) and 
again in the absence of the test compound (Sequence C). 
Receptor tyrosine kinase inhibition was calculated as follows: 
##EQU1## 
The extent of inhibition was then determined at a range of concentrations 
of test compound to give an IC.sub.50 value. 
(b) An in vitro assay which determines the ability of a test compound to 
inhibit the EGF-stimulated growth of the human naso-pharyngeal cancer cell 
line KB. 
KB cells were seeded into wells at a density of 1.times.10.sup.4 
-1.5.times.10.sup.4 cells per well and grown for 24 hours in DMEM 
supplemented with 5% FCS (charcoal-stripped). Cell growth was determined 
after incubation for 3 days by the extent of metabolism of MTT tetrazolium 
dye to furnish a bluish colour. Cell growth was then determined in the 
presence of EGF (10 ng/ml) or in the presence of EGF (10 ng/ml) and a test 
compound at a range of concentrations. An IC.sub.50 value could then be 
calculated. 
(c) An in-vivo assay in a group of athymic nude mice (strain ONU:Alpk) 
which determines the ability of a test compound (usually administered 
orally as a ball-milled suspension in 0.5% polysorbate) to inhibit the 
growth of xenografts of the human vulval epidermoid carcinoma cell line 
A-431. 
A-431 cells were maintained in culture in DMEM supplemented with 5% FCS and 
2 mM glutamine. Freshly cultured cells were harvested by trypsinization 
and injected subcutaneously (10 million cells/0.1 ml/mouse) into both 
flanks of a number of donor nude mice. When sufficient tumour material was 
available (after approximately 9 to 14 days), fragments of tumour tissue 
were transplanted in the flanks of recipient nude mice (test day 0). 
Generally, on the seventh day after transplantation (test day 7) groups of 
7 to 10 mice with similar-sized tumours were selected and dosing of the 
test compound was commenced. Once daily dosing of test compound was 
continued for a total of 13 days (test days 7 to 19 inclusive). In some 
studies the dosing of the test compound was continued beyond test day 19, 
for example to test day 26. In each case, on the following test day the 
animals were killed and the final tumour volume was calculated from 
measurements of the length and width of the tumours. Results were 
calculated as a percentage inhibition of tumour volume relative to 
untreated controls. 
The compounds of the inventions in general, possess activity at 
approximately the following concentrations or doses in tests (a) to (c): 
______________________________________ 
Test (a):- 
IC.sub.50 in the range, for example, 0.001 to 1 .mu.M; 
Test (b):- 
IC.sub.50 in the range, for example, 0.1 to 1 .mu.M; 
Test (c):- 
20 to 90% inhibition of tumour volume from a daily dose in 
the range, for example, 12.5 to 200 mg/kg. 
______________________________________ 
The quinazoline derivative 
4-(3'-chloro-4'-fluoroanilino)-6-(3-diethylaminopropoxy)quinazoline 
possesses activity at approximately the following concentrations or 
doses:--Test (a): IC.sub.50 0.003 .mu.M; Test (b): IC.sub.50 0.1 .mu.M; 
Test (c): greater than 50% inhibition of tumour volume from a daily dose 
of 50 mg/kg. 
The quinazoline derivative 
4-(3',4'-difluoroanilino)-6-(3-morpholinopropoxy)quinazoline possesses 
activity at approximately the following concentrations or doses: Test (b): 
IC.sub.50 1.2 .mu.M; Test (c): greater than 50% inhibition of tumour 
volume from a daily dose of 50 mg/kg. 
According to a further aspect of the invention there is provided a 
pharmaceutical composition which comprises a quinazoline derivative of the 
formula I, or a pharmaceutically-acceptable salt thereof, in association 
with a pharmaceutically-acceptable diluent or carrier. 
The composition may be in a form suitable for oral administration, for 
example as a tablet or capsule, for parenteral injection (including 
intraveous, subcutaneous, intramuscular, intravascular or infusion) as a 
sterile solution, suspension or emulsion, for topical administration as an 
ointment or cream or for rectal administration as a suppository. 
In general the above compositions may be prepared in a conventional manner 
using conventional excipients. 
The quinazoline derivative will normally be administered to a warm-blooded 
animal at a unit dose within the range 5-10000 mg per square meter body 
area of the animal, i.e. approximately 0.1-200 mg/kg, and this normally 
provides a therapeutically-effective dose. A unit dose form such as a 
tablet or capsule will usually contain, for example 1-250 mg of active 
ingredient. Preferably a daily dose in the range of 1-100 mg/kg is 
employed. However the daily dose will necessarily be varied depending upon 
the host treated, the particular route of administration, and the severity 
of the illness being treated. Accordingly the optimum dosage may be 
determined by the practitioner who is treating any particular patient. 
According to a further aspect of the present invention there is provided a 
quinazoline derivative of the formula I, or a pharmaceutically-acceptable 
salt thereof, for use in a method of treatment of the human or animal body 
by therapy. 
We have now found that the compounds of the present invention possess 
anti-proliferative properties such as anti-cancer properties which are 
believed to arise from their Class I receptor tyrosine kinase inhibitory 
activity. Accordingly the compounds of the present invention are expected 
to be useful in the treatment of diseases or medical conditions mediated 
alone or in part by Class I receptor tyrosine kinases, i.e. the compounds 
may be used to produce a Class I receptor tyrosine kinase inhibitory 
effect in a warm-blooded animal in need of such treatment. Thus the 
compounds of the present invention provide a method for treating the 
proliferation of malignant cells characterised by inhibition of Class I 
receptor tyrosine kinases, i.e. the compounds may be used to produce an 
anti-proliferative effect mediated alone or in part by the inhibition of 
Class I receptor tyrosine kinase. Accordingly the compounds of the present 
invention are expected to be useful in the treatment of psoriasis and/or 
cancer by providing an anti-proliferative effect, particularly in the 
treatment of Class I receptor tyrosine kinase sensitive cancers such as 
cancers of the breast, lung, colon, rectum, stomach, prostate, bladder, 
pancreas and ovary. 
Thus according to this aspect of the invention there is provided the use of 
a quinazoline derivative of the formula I, or a 
pharmaceutically-acceptable salt thereof, in the manufacture of a 
medicament for use in the production of an anti-proliferative effect in a 
warm-blooded animal such as man. 
According to a further feature of this aspect of the invention there is 
provided a method for producing an anti-proliferative effect in a 
warm-blooded animal, such as man, in need of such treatment which 
comprises administering to said animal an effective amount of a 
quinazoline derivative as defined immediately above. 
As stated above the size of the dose required for the therapeutic or 
prophylactic treatment of a particular proliferative disease will 
necessarily be varied depending on the host treated, the route of 
administration and the severity of the illness being treated. A unit dose 
in the range, for example, 1-200 mg/kg, preferably 1-100 mg/kg is 
envisaged. 
The anti-proliferative treatment defined hereinbefore may be applied as a 
sole therapy or may involve, in addition to the quinazoline derivative of 
the invention, one or more other anti-tumour substances, for example those 
selected from, for example, mitotic inhibitors, for example vinblastine, 
vindesine and vinorelbine; tubulin disassembly inhibitors such as taxol; 
alkylating agents, for example cis-platin, carboplatin and 
cyclophosphamide; antimetabolites, for example 5-fluorouracil, tegafur, 
methotrexate, cytosine arabinoside and hydroxyurea, or, for example, one 
of the preferred antimetabolites disclosed in European Patent Application 
No. 239362 such as 
N-{5-N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-methylamino!-2 
-thenoyl}-L-glutamic acid; intercalating antibiotics, for example 
adriamycin, mitomycin and bleomycin; enzymes, for example asparaginase; 
topoisomerase inhibitors, for example etoposide and camptothecin; 
biological response modifiers, for example interferon; and anti-hormones, 
for example antioestrogens such as tamoxifen or, for example antiandrogens 
such as 
4'-cyano-3-(4-fluorophenylsulphonyl)-2-hydroxy-2-methyl-3'-(trifluoromethy 
l)propionanilide. Such conjoint treatment may be achieved by way of the 
simultaneous, sequential or separate dosing of the individual components 
of the treatment. According to this aspect of the invention there is 
provided a pharmaceutical product comprising a quinazoline derivative of 
the formula I as defined hereinbefore and an additional anti-tumour 
substance as defined hereinbefore for the conjoint treatment of cancer. 
As stated above the quinazoline derivative defined in the present invention 
is an effective anti-cancer agent, which property is believed to arise 
from its Class I receptor tyrosine kinase inhibitory properties. Such a 
quinazoline derivative of the invention is expected to possess a wide 
range of anti-cancer properties as Class I receptor tyrosine kinases have 
been implicated in many common human cancers such as leukaemia and breast, 
lung, colon, rectal, stomach, prostate, bladder, pancreas and ovarian 
cancer. Thus it is expected that the quinazoline derivative of the 
invention will possess anti-cancer activity against these cancers. It is 
in addition expected that the quinazoline derivative of the present 
invention will possess activity against a range of leukaemias, lymphoid 
malignancies and solid tumours such as carcinomas and sarcomas in tissues 
such as the liver, kidney, prostate and pancreas. 
It is further expected that a quinazoline derivative of the invention will 
possess activity against other diseases involving excessive cellular 
proliferation such as psoriasis, benign prostatic hypertrophy (BPH) and 
atherosclerosis. 
It is also to be expected that a quinazoline derivative of the invention 
will be useful in the treatment of additional disorders of cellular growth 
in which aberrant cell signalling by way of receptor tyrosine kinase 
enzymes, including as yet unidentified receptor tyrosine kinase enzymes, 
are involved. Such disorders include, for example, inflammation, 
angiogenesis, vascular restenosis, immunological disorders, pancreatitis, 
kidney disease and blastocyte maturation and implantation.