EA042928B1 - POLYMORPHIC FORM OF N-(2-{2-DIMETHYLAMINOETHYLMETHYLAMINO}-4-METHOXY-5-{[4-(1-METHYLINDOL-3-YL)PYRIMIDIN-2-YL]AMINO}PHENYL)PROP-2-ENAMIDE MESYLATE SALT - Google Patents

Description

The invention relates to a polymorphic form of the mesylate salt of N(2-{2-dimethylaminoethylmethylamino}4-methoxy-5-{[4-(1-methylindol-3-yl)pyrimidin-2-yl]amino}phenyl)prop-2-enamide . Said polymorph may be useful in the treatment or prevention of a disease or medical condition mediated by certain mutated forms of the epidermal growth factor receptor (eg, L858R activating mutant, exon 19 deletion activating mutant, and T790M resistant mutant). Such compounds and their salts may be useful in the treatment or prevention of a number of different cancers. The invention also relates to pharmaceutical compositions containing said compounds and salts thereof, particularly useful polymorphs of said compounds and salts, intermediates useful in the preparation of said compounds, and methods of treating diseases mediated by various different forms of EGFR using said compounds and their salts.

EGFR is a transmembrane tyrosine kinase protein that is a member of the erbB receptor family. Upon binding a growth factor ligand such as epidermal growth factor (EGF), the receptor may homodimerize with another EGFR molecule or heterodimerize with another family member such as erbB2 (HER2), erbB3 (HER3) or erbB4 (HER4).

Homo- and/or heterodimerization of erbB receptors results in phosphorylation of key tyrosine residues in the intracellular domain and leads to stimulation of a wide variety of intracellular signaling pathways involved in cell proliferation and survival. Dysregulation of erbB family signaling triggers proliferation, invasion, metastasis, angiogenesis, and tumor cell survival and has been described for many human cancers, including lung, head and neck, and breast.

Thus, the ErbB family represents a rational target for anticancer drug development, and a number of agents targeting EGFR or erbB2 are currently available in clinical practice, including gefitinib (IRESSA™), erlotinib (TARCEVA™), and lapatinib (TYKERB™, TYVERB ™). A detailed description of erbB receptor signaling and its involvement in tumorigenesis is presented in the New England Journal of Medicine (2008) Vol. 358, 1160-74 and Biochemical and Biophysical Research Communications (2004) Vol. 319, 1-11.

It was reported in 2004 (Science [2004] Vol. 304, 1497-500, and New England Journal of Medicine [2004] Vol. 350, 2129-39) that activating mutations in EGFR correlated with response to gefitinib therapy in non-small cell cancer. lungs (NSCLC). The most common EGFR activating mutations L858R and delE746_A750 result in increased affinity for small molecule tyrosine kinase inhibitors such as gefitinib and erlotinib and decreased affinity for adenosine triphosphate (ATP) compared to wild-type (WT) EGFR. As a result, acquired resistance to gefitinib or erlotinib therapy occurs, for example, through mutation of the T790M residue gatekeeper gene, which is reported to be found in 50% of clinically resistant patients. It is believed that this mutation does not prevent gefitinib or erlotinib from binding to EGFR sterically, but merely alters ATP affinity to levels comparable to WT EGFR.

In light of the importance of this mutation in resistance to existing therapies directed to EGFR, the authors believe that agents that can inhibit an EGFR-carrying pyloric gene mutation may be particularly useful in the treatment of cancer.

There remains a need for compounds that can exhibit favorable efficacy profiles against WT EGFR when compared to activating EGFR mutants (e.g. L858R EGFR mutant, or delE746_A750 mutant, or exon 19 deletion EGFR mutant), and/or resistant EGFR mutants ( EGFR mutant T790M), and/or selectively for other enzymatic receptors, which may make the compounds particularly promising for development as therapeutic agents. In this regard, there remains a need for compounds that show a higher degree of inhibition of certain activating or resistance mutants of EGFR while showing relatively low inhibition of WT EGFR. Such compounds are expected to be more suitable as therapeutic agents, especially for the treatment of cancer, in view of reducing the toxicological symptoms associated with WT EGFR inhibition. Such toxicological signs are known to manifest as such in humans in the form of skin rashes and/or diarrhea. Applicants have surprisingly found that one or more 2-(2,4,5-substituted anilino)pyrimidine compounds are highly effective against several EGFR mutants while exhibiting relatively low inhibition of WT EGFR.

The compound(s) of the invention may also exhibit favorable physical properties (eg high water solubility, high permeability and/or low plasma protein binding) and/or favorable toxicity profiles (eg reduced hERG blocking capacity) and/or favorable metabolic properties. profiles compared to other known inhibitors of EGFR/EGFR mutants. Thus, such compound(s) may be particularly useful(s) in the treatment of disease states that involve EGFR and/or EGFR activating mutations and/or EGFR resistance mutations, for example in the treatment of cancer.

- 1 042928

Brief summary of the invention

The present invention relates to a polymorphic form of N-(2-{2-dimethylaminoethylmethylamino}-4-methoxy-5-{[4-(1-methylindol-3-yl)pyrimidin-2-yl]amino}phenyl)prop- 2-enamide, which has an X-ray powder diffraction pattern with characteristic peaks at 2-theta values of 7.2; 8.6; 9.5; 10.4; 15.3; 16.4; 18.8; 22.1; 25.7 and 26.1°, where indicated values may differ by plus or minus 0.2° 2-theta measured using CuKa.

Detailed description of the invention

The invention provides a compound of formula (I)

R (I), where G is selected from 4,5,6,7-tetrahydropyrazolo[1,5-a]pyridin-3-yl, 1H-indol-3-yl, 1-methyl-1Hindol-3-yl and pyrazolo [1,5-a]pyridin-3-yl;

R ¹ is selected from hydrogen, fluorine, chlorine, methyl and cyano;

R ² is selected from methoxy and methyl and

R ³ is selected from (3R)-3-(dimethylamino)pyrrolidin-1-ylα, (3S)-3-(dimethylamino)pyrrolidin-1-yl, 3(dimethylamino)azetidin-1-yl, [2-(dimethylamino) ethyl]-(methyl)amino, [2-(methylamino)ethyl](methyl)amino, 5-methyl-2,5-diazaspiro[3.4]oct-2-yl, (3aR,6aR)-5-methylhexa-hydropyrrolo [3,4-b]pyrrol-1(2H)-yl, 1-methyl-1,2,3,6-tetrahydropyridin-4-yl, 4-methylpiperizin-1-yl, 4-[2-(dimethylamino) -2oxoethyl]piperazin-1-yl, methyl [2-(4-methylpiperazin-1-yl)ethyl]amino, methyl [2-(morpholin-4yl)ethyl]amino, 1-amino-1,2,3,6 -tetrahydropyridin-4-yl and 4-[(28)-2-aminopropanoyl]piperazin-1-yl; or a pharmaceutically acceptable salt thereof.

A suitable pharmaceutically acceptable salt of a compound of formula (I) is, for example, an acid addition salt. For example, an acid addition salt can be formed using an inorganic or organic acid. An acid addition salt may be formed using an inorganic acid selected from hydrochloric acid, hydrobromic acid, sulfuric acid, and phosphoric acid. The acid addition salt may be formed using an organic acid selected from trifluoroacetic acid, citric acid, maleic acid, oxalic acid, acetic acid, formic acid, benzoic acid, fumaric acid, succinic acid, tartaric acid, lactic acid, pyruvic acid, methanesulfonic acid acid, benzenesulfonic acid and p-toluenesulfonic acid.

In one embodiment, N-(2-{2-dimethylaminoethylmethylαmino}-4methoxy-5-{[4-(1-methylindol-3-yl)pyrimidin-2-yl]amino}phenyl)prop-2-enamide mesylate salt is provided.

It should be understood that the compound of formula (I) and its pharmaceutically acceptable salts may exist in solvated forms and unsolvated forms. For example, the solvated form may be the hydrated form. It is to be understood that the present invention encompasses all such solvated and unsolvated forms.

The compound of formula (I) may be administered in the form of a prodrug which is degraded in the human or animal body to give the compound of formula (I). Examples of prodrugs include in vivo hydrolysable esters of the compound of formula (I). In vivo hydrolysable esters can be formed by esterification of the hydroxyl group in a compound of formula (I). Various forms of prodrugs are known in the art. For example, such prodrugs can be found in:

a) Design of Prodrugs, edited by H. Bundgaard, (Elsevier, 1985), and Methods in Enzymology, Vol. 42, p. 309-396, edited by K. Widder, et al. (Academic Press, 1985);

b) A Textbook of Drug Design and Development, edited by Krogsgaard-Larsen and H. Bundgaard, Chapter 5 Design and Application of Prodrugs, by H. Bundgaard p. 113-191 (1991);

c) H. Bundgaard, Advanced Drug Delivery Reviews, 8, 1-38 (1992);

d) H. Bundgaard, et al., Journal of Pharmaceutical Sciences, 77, 285 (1988); and N. Kakeya, et al, Chem Pharm Bull, 32, 692 (1984).

In one aspect of the invention, there are provided compounds of formula (I) which inhibit one or more EGFR activating or resistance mutations, eg L858R activating mutant, exon 19 deletion EGFR activating mutant, and T790M resistance mutant. Advantageously, such compounds may be useful in the treatment of cancer in a patient who has developed or may be at risk of developing a level of resistance to the level of existing EGFR inhibitor therapies.

In one aspect, compounds of formula (I) are provided which exhibit a higher degree

- 2 042928 inhibition of activating or resistance mutants of EGFR compared to WT EGFR. Such compounds are expected to be more suitable as therapeutic agents, especially for the treatment of cancer, in view of reducing the toxicological symptoms associated with WT EGFR inhibition. Such toxicological signs are known to manifest as such in humans in the form of skin rashes and/or diarrhea.

Compounds of formula (I) can be prepared by reacting a compound of formula (II)

R (P) or salts thereof (wherein G, R ¹ , R ² and R ³ are as defined herein), with an activated acrylic acid derivative (for example, acryloyl chloride or an appropriate activated ester), in a solvent such as CH ₂ Cl ₂ , tetrahydrofuran, N,N-dimethylformamide or N,N-dimethylacetamide. Thus, the intermediates of formula (II) form a further aspect of the present invention.

Thus, in a further aspect of the invention, there is provided a compound of formula (II) (as shown above), wherein

G is selected from 1H-indol-3-yl, 1-methyl-1H-indol-3-yl and pyrazolo[1,5-a]-pyridin-3-yl;

R ¹ is selected from hydrogen, chlorine, methyl and cyano;

R ² is methoxy and

R ³ is selected from (3R)-3-(dimethylamino)pyrrolidin-1-yl, (3S)-3-(dimethylamino)pyrrolidin-1-yl, 3(dimethylamino)azetidin-1-yl, [2-(dimethylamino) ethyl]-(methyl)amino, 5-methyl-2,5-diazaspiro[3.4]oct2-yl, (3aR,6aR)-5-methylhexa-hydropyrrolo[3,4-b]pyrrol-1(2H)-yl , 1-methyl-1,2,3,6-tetrahydropyridin-4-yl, 4-methylpiperizin-1-yl, 4-[2-(dimethylamino)-2-oxoethyl]piperazin-1-yl, methyl[2- (4methylpiperazin-1-yl)ethyl]amino, methyl[2-(morpholin-4-yl)ethyl]amino, 1-amino-1,2,3,6-tetrahydropyridin-4-yl and 4-[(2S) -2-(t-butoxycarbonyl)aminopropanoyl]piperazin-1-yl; or its salt.

In further embodiments, the invention provides any of the intermediates (as they are named here in the experimental section, in their free base form).

In further embodiments, the invention provides any of the intermediates (as referred to herein in the experimental section, in their free base form) or their pharmaceutically acceptable salts.

Thus, examples of just some of the above embodiments are given below:

In one embodiment, intermediate 100 or a salt thereof is provided.

Thus, in this case, N ¹ -(2-dimethylaminoethyl)-5-methoxy-N ¹ -methyl-N ⁴ -[4-(1methylindol-3-yl)pyrimidin-2-yl]benzene-1,2, 4-triamine or its salt.

One embodiment provides N ¹ -(2-dimethylaminoethyl)-5-methoxy-N ¹ -methyl-N ⁴ -[4-(1methylindol-3-yl)pyrimidin-2-yl]benzene-1,2,4-triamine .

In one embodiment, intermediate 168 or a salt thereof is provided.

Thus, in this case, N ¹ -(2-dimethylaminoethyl)-N ⁴ -[4-(1H-indol-3yl)pyrimidin-2-yl]-5-methoxy-N ¹ -methylbenzene-1,2,4 -triamine or its salt.

One embodiment provides N ¹ -(2-dimethylaminoethyl)-N ⁴ -[4-(1H-indol-3-yl)pyrimidin-2-yl]5-methoxy-N ¹ -methylbenzene-1,2,4-triamine .

Amino compounds of formula (II) can be obtained by reduction of the corresponding nitro compounds. When G is indol-3-yl, then the nitrogen atom of the indole group may be protected by a suitable nitrogen protecting group, for example a phenylsulfonyl protecting group. For examples of protecting groups, including protecting groups suitable for protecting nitrogen atoms (as well as means of formation and possible deprotection), see T.W. Greene and P.G.M. Wuts, Protective Groups in Organic Synthesis, Second Edition, John Wiley & Sons, New York, 1991.

These additional intermediates form another aspect of the invention.

Thus, in a further aspect of the invention, there is provided a compound of formula (III)

(ΠΙ), where G is selected from 1H-indol-3-yl, 1-methyl-1H-indol-3-yl and 1-(N-protective group)-indol-3-yl and pyrazolo[1,5-a ]pyridin-3-yl;

R ¹ is selected from hydrogen, chlorine, methyl and cyano;

- 3 042928

R ² is methoxy and

R ³ is selected from (3R)-3-(dimethylamino)pyrrolidin-1-yl, (3S)-3-(dimethylamino)-pyrrolidin-1-yl, 3(dimethylamino)azetidin-1-yl, [2-(dimethylamino )ethyl]-(methyl)amino, 5-methyl-2,5-diazaspiro[3.4]oct-2-yl, (3 aR,6aR)-5-methylhexa-hydropyrrolo[3,4-b]pyrrole- 1 ( 2H)-yl, 1-methyl-1,2,3,6-tetrahydropyridin-4-yl, 4methylpiperizin-1-yl, 4-[2-(dimethylamino)-2-oxoethyl]piperazin-1-yl, methyl[ 2-(4-methylpiperazin-1yl)ethyl]amino, methyl[2-(morpholin-4-yl)ethyl]amino, 1-amino-1,2,3,6-tetrahydropyridin-4-yl and 4-[( 28)-2-(tert-butoxycarbonyl)aminopropanoyl]piperazin-1-yl; or its salt.

One example of an N-protecting group in the 1-(N-protecting group)-indol-3-yl radical is a phenylsulfonyl group.

In one embodiment, intermediate 101 or a salt thereof is provided.

Thus, in this case, N'-(2-dimethylaminoethyl)-2-methoxy-N'-methyl-N-[4-(1methylindol-3-yl)pyrimidin-2-yl]-5-nitrobenzene-1, 4-diamine or its salt.

One embodiment provides N'-(2-dimethylaminoethyl)-2-methoxy-N'-methyl-N-[4-(1methylindol-3-yl)pyrimidin-2-yl]-5-nitrobenzene-1,4-diamine .

In one embodiment, intermediate 169 or a salt thereof is provided.

Thus, in this case, N'-(2-dimethylaminoethyl)-N-[4-(1H-indol-3yl)pyrimidin-2-yl]-2-methoxy-N'-methyl-5-nitrobenzene-1, 4-diamine or its salt.

In one embodiment, N'-(2-dimethylaminoethyl)-N-[4-(1H-indol-3-yl)pyrimidin-2-yl]2-methoxy-N'-methyl-5-nitrobenzene-1,4- diamine.

In one embodiment, intermediate 175 or a salt thereof is provided.

Thus, in this case, N ⁴ -(2-dimethylaminoethyl)-2-methoxy-N ⁴ -methyl-N ¹ -[4-(1methylindol-3-yl)pyrimidin-2-yl]-5-nitro-benzene -1,4-diamine or its salt.

In one embodiment, N ⁴ -(2-dimethylaminoethyl)-2-methoxy-N ⁴ -methyl-N ¹ -[4-(1methylindol-3-yl)pyrimidin-2-yl]-5-nitro-benzene-1 is provided, 4-diamine.

Some compounds of formula (III) have an R ³ group attached to the phenyl ring of formula (III) through the nitrogen atom of the R ³ group. These compounds can be prepared by reacting the appropriate amine with the appropriate fluoro compound. Such intermediates form another aspect of the invention.

Thus, in a further aspect of the invention, there is provided a compound of formula (IV)

(IV) wherein G is selected from 1H-indol-3-yl, 1-methyl-1H-indol-3-yl and pyrazolo[1,5-a]pyridin-3-yl;

R ¹ is selected from hydrogen, chlorine, methyl and cyano and

R ² is methoxy; or its salt.

In one embodiment of this aspect of the invention, there is provided a compound of formula (IV) as shown above, wherein

G is selected from 4,5,6,7-tetrahydropyrazolo[1,5-a]pyridin-3-yl, 1H-indol-3-yl, 1-methyl-1H-indol-3yl and pyrazolo[1,5-a ]pyridin-3-yl;

R ¹ is selected from hydrogen, chlorine, methyl and cyano and

R ² is methoxy; or its salt.

In one embodiment, intermediate 68 or a salt thereof is provided.

Thus, N-(4-fluoro-2-methoxy-5-nitrophenyl)-4-(1H-indol-3yl)pyrimidin-2-amine or a salt thereof is provided here.

In one embodiment, N-(4-fluoro-2-methoxy-5-nitrophenyl)-4-(1H-indol-3-yl)pyrimidin-2-amine is provided.

In one embodiment, intermediate 129 or a salt thereof is provided.

Thus, N-(4-fluoro-2-methoxy-5-nitrophenyl)-4-(1-methylindol3-yl)pyrimidin-2-amine or a salt thereof is provided here.

In one embodiment, N-(4-fluoro-2-methoxy-5-nitrophenyl)-4-(1-methylindol-3yl)pyrimidine-2-amine is provided.

In one embodiment, intermediate 176 or a salt thereof is provided.

Thus, N-(4-fluoro-2-methoxy-5-nitrophenyl)-4-(1-methylindol3-yl)pyrimidin-2-amine or a salt thereof is provided here.

In one embodiment, N-(4-fluoro-2-methoxy-5-nitrophenyl)-4-(1-methylindol-3yl)pyrimidine-2-amine is provided.

Some compounds of formula (III) have an R ³ group attached to the phenyl ring

- 4 042928 formula (III) through the nitrogen atom of the group R ³ . These compounds can be prepared by reacting an appropriate organoboron compound (eg, a borate ester compound) with a suitable aryl bromide or aryl chloride compound. For example, the organoboron compound may be (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-R ³ . Such intermediates form another aspect of the invention.

Thus, in a further aspect of the invention, there is provided a compound of formula (V)

where X represents bromine or chlorine;

G is selected from 4,5,6,7-tetrahydropyrazolo[1,5-a]pyridin-3-yl, 1H-indol-3-yl, 1-methyl-1H-indol-3yl and pyrazolo[1,5-a ]pyridin-3-yl;

R ¹ is selected from hydrogen, chlorine, methyl and cyano and

R ² is methoxy; or its salt.

In one embodiment of a compound of formula (V), X is bromine.

In one embodiment of the compound of formula (V), X is chlorine.

In one embodiment, intermediate 145 is provided.

Thus, N-(4-bromo-2-methoxy-5-nitrophenyl)-4-(1-methylindol3-yl)pyrimidin-2-amine or a salt thereof is provided here.

In one embodiment, N-(4-bromo-2-methoxy-5-nitrophenyl)-4-(1-methylindol-3yl)pyrimidine-2-amine is provided.

Preferably, the compounds of the present invention can be prepared by reacting a compound of formula (II) with 3-chloropropanoyl chloride in the presence of a base (eg an alkali metal carbonate, eg potassium carbonate) in a suitable solvent, eg acetone. In this way, an intermediate compound of formula (VI) is obtained, which can be isolated as a solid (in its free base form or as a salt), or it can remain in solution, and treated with a base (for example, an alkali metal hydroxide, for example NaOH) for converting a compound of formula (VI) into the corresponding compound of formula (I). Thus, compounds of formula (VI) and salts thereof are useful chemical intermediates in the formation of compounds of formula (I). Thus, in a further aspect of the invention, there is provided a compound of formula (VI)

or its salt, where G, R ¹ , R ² and R ³ are as defined here.

In one embodiment, there is provided a compound of formula (VI) as shown above, or a salt thereof, wherein

G is selected from 4,5,6,7-tetrahydropyrazolo[1,5-a]pyridin-3-yl, 1H-indol-3-yl, 1-methyl-1H-indol-3yl and pyrazolo[1,5-a ]pyridin-3-yl;

R ¹ is selected from hydrogen, fluorine, chlorine, methyl and cyano;

R ² is selected from methoxy and methyl and

R ³ is selected from (3R)-3-(dimethylamino)pyrrolidin-1-yl, (3S)-3-(dimethylamino)pyrrolidin-1-yl, 3(dimethylamino)azetidin-1-yl, [2-(dimethylamino) ethyl]-(methyl)amino, [2-(methylamino)ethyl](methyl)amino, 5-methyl-2,5-diazaspiro[3.4]oct-2-yl, (3aR,6aR)-5-methylhexa-hydropyrrolo [3,4-b]pyrrole1(2I)-yl, 1-methyl-1,2,3,6-tetrahydropyridin-4-yl, 4-methylpiperizin-1-yl, 4-[2-(dimethylamino)-2oxoethyl ]piperazin-1-yl, methyl[2-(4-methylpiperazin-1-yl)ethyl]amino, methyl[2-(morpholin-4yl)ethyl]amino, 1-amino-1,2,3,6-tetrahydropyridine -4-yl; and 4-[(2S)-2-aminopropanoyl]piperazin-1-yl.

In a further embodiment, there is provided a compound of formula (VI) as shown above, or a salt thereof, wherein

G is selected from 1H-indol-3-yl, 1-methyl-1H-indol-3-yl and pyrazolo[1,5-a]-pyridin-3-yl;

R ¹ is selected from hydrogen, fluorine, chlorine, methyl and cyano;

R ² is selected from methoxy and methyl and

R ³ is selected from (3R)-3-(dimethylamino)pyrrolidin-1-yl, (3S)-3-(dimethylamino)pyrrolidin-1-yl, 3-5 042928 (dimethylamino)azetidin-1-yl, [2- (dimethylamino)ethyl]-(methyl)amino, 5-methyl-2,5-diazaspiro[3.4]oct2-yl, (3aR,6aR)-5-methylhexa-hydropyrrolo[3,4-b]pyrrole-1(2H )-yl, 1-methyl-1,2,3,6-tetrahydropyridin-4-yl, 4-methylpiperizin-1-yl, 4-[2-(dimethylamino)-2-oxoethyl]piperazin-1-yl, methyl [2-(4methylpiperazin-1-yl)ethyl]amino, methyl[2-(morpholin-4-yl)ethyl]amino, 1-amino-1,2,3,6-tetrahydropyridin-4-yl and 4-[ (2S)-2-aminopropanoyl]piperazin-1-yl.

In a further embodiment, intermediate 174 or a salt thereof is provided.

Thus, in this case, proposed 3-chloro-N-[2-[2-dimethylaminoethyl-(methyl)amino]-4methoxy-5-[[4-(1-methylindol-3-yl)pyrimidin-2-yl] amino]phenyl]propanamide or a salt thereof.

In one embodiment, 3-chloro-K-[2-[2-dimethylaminoethyl-(methyl)amino]-4-methoxy-5-[[4(1-methylindol-3-yl)pyrimidin-2-yl]amino] is provided. phenyl]propanamide.

Other intermediates are useful in the preparation of certain compounds of formula (I). Thus, for example, in one embodiment, intermediate 170 or a salt thereof is provided. In a further embodiment, intermediate 171 or a salt thereof is provided. In yet another embodiment, intermediate 172 or a salt thereof is provided. In one embodiment, intermediate 144 or a salt thereof is provided.

According to a further aspect of the invention, there is provided a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, as defined above, together with a pharmaceutically acceptable diluent or carrier.

The compositions of the invention may be presented in a form suitable for oral administration (for example, tablets, lozenges, hard or soft capsules, aqueous or oily suspensions, emulsions, dispersible powders or granules, syrups or elixirs), for topical administration (for example, in the form of creams, ointments, gels or aqueous or oily solutions or suspensions), for administration by inhalation (for example, as a finely divided powder or liquid aerosol), for administration by insufflation (for example, as a finely divided powder), or for parenteral administration (for example , as a sterile aqueous or oily solution for intravenous, subcutaneous, intramuscular or intramuscular dosing or as a suppository for rectal dosing).

Compositions of the invention may be prepared by conventional techniques using conventional pharmaceutical excipients well known in the art. Thus, compositions intended for oral administration may contain, for example, one or more colorants, sweeteners, flavoring agents and/or preservatives.

The compound of formula (I) is typically administered to a warm-blooded animal at a single dose in the range of 5-5000 mg/m ² body area of the animal, that is, about 0.1-100 mg/kg, and this usually provides a therapeutically effective dose. A unit dosage form, such as a tablet or capsule, typically contains, for example, 1-250 mg of the active ingredient. The daily dose will vary as needed depending on the organism to be treated, the particular route of administration, and the severity of the disease being treated. Accordingly, a physician treating a particular patient can determine the optimal dosage.

In the context of the present description, the term therapy also includes prophylaxis, unless there are specific indications to the contrary. The terms therapeutic and therapeutic should be interpreted accordingly.

The term treatment, as used herein, has its usual everyday meaning in the fight against a disease with the aim of completely or partially alleviating some or all of its symptoms, or correcting or compensating for the pathology underlying it.

The term prophylaxis, as used herein, has a common everyday meaning and includes primary prevention of the development of a disease and secondary prevention, in which, if the disease has already developed, the patient is temporarily or permanently protected from exacerbation or worsening of the disease or the development of new symptoms associated with this disease.

As a result of its inhibitory activity against L858R EGFR mutant, T790M EGFR mutant, and exon 19 deletion activating mutant, the compound of formula (I) and pharmaceutically acceptable salts thereof are expected to be useful in the treatment of diseases or medical conditions mediated solely or in part by EGFR. mutant activity, such as cancer. Cancers that can be treated using a compound of formula (I) or a pharmaceutically acceptable salt thereof include ovarian cancer, cervical cancer, colorectal cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, melanoma, prostate cancer, leukemia, lymphoma, non-Hodgkin's lymphoma, gastric cancer, lung cancer, hepatocellular carcinoma, gastric cancer, gastrointestinal stromal tumor (GIST), thyroid cancer, bile duct cancer, endometrial cancer, kidney cancer, anaplastic large cell lymphoma, acute myeloid leukemia (AML ), but not limited to multiple myeloma, melanoma, and mesothelioma.

It is envisaged that in the methods of treating cancer mentioned herein, the compound of formula (I) will be administered to a mammal, more specifically a human. Similarly, in the case of using the Compound of formula (I) for the treatment of cancer mentioned herein, it is envisaged that the compound of formula (I) will be administered to a mammal, more specifically a human.

According to another aspect of the invention, there is provided a compound of formula (I) as defined herein, or a pharmaceutically acceptable salt thereof, for use as a medicament.

According to a further aspect of the invention, there is provided a compound of formula (I) as defined herein, or a pharmaceutically acceptable salt thereof, for use in the treatment of a disease mediated by an EGFR L858R mutant and/or a T790M EGFR and/or exon 19 deletion activating mutant. In one In an embodiment of the invention, said disease mediated through an EGFR L858R mutant and/or a T790M EGFR and/or exon 19 deletion activating mutant is a cancer.

According to a further aspect of the invention, there is provided the use of a compound of formula (I) as defined herein, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of a disease mediated by an EGFR L858R mutant and/or an EGFR T790M mutant and/or an exon 19 deletion activating mutant. In one embodiment of the invention, said disease mediated through an EGFR L858R mutant and/or a T790M EGFR and/or exon 19 deletion activating mutant is a cancer.

According to a further aspect of the invention, the use of a compound of formula (I) as defined herein, or a pharmaceutically acceptable salt thereof, is provided for the manufacture of a medicament for the treatment of cancer.

According to a further aspect of the invention, there is provided a method of using a compound of formula (I) as defined herein, or a pharmaceutically acceptable salt thereof, for the treatment of cancer.

According to this aspect of the invention, there is provided a method for producing an anti-cancer effect in a warm-blooded animal, such as a human, in need of such treatment, comprising administering to said animal an effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, as defined herein.

According to a further aspect of the invention, there is provided a method of treating a human afflicted with a disease wherein inhibition of an L858R EGFR mutant and/or a T790M EGFR mutant and/or an exon 19 deletion activating mutant is beneficial, comprising the steps of administering to a subject in need thereof a therapeutically effective amount of a compound of the formula (I) as defined above, or a pharmaceutically acceptable salt thereof. In one embodiment of the invention, the disease benefiting from inhibition of an L858R EGFR mutant and/or a T790M EGFR mutant and/or exon 19 deletion activating mutant is cancer.

In any of the aspects or embodiments mentioned herein, where cancer is referred to in a general sense, said cancer may be selected from ovarian cancer, cervical cancer, colorectal cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, melanoma, prostate cancer cancer, leukemia, lymphoma, non-Hodgkin's lymphoma, gastric cancer, lung cancer, hepatocellular carcinoma, gastric cancer, gastrointestinal stromal tumor (GIST), thyroid cancer, bile duct cancer, endometrial cancer, kidney cancer, anaplastic large cell lymphoma, acute myeloid leukemia (AML), multiple myeloma, melanoma and mesothelioma.

In any of the aspects or embodiments of the invention where cancer is referred to in a general sense, the following embodiments may be applicable.

In one embodiment, the cancer is ovarian cancer.

In one embodiment, the cancer is cervical cancer.

In one embodiment, the cancer is colorectal cancer.

In one embodiment, the cancer is breast cancer.

In one embodiment, the cancer is pancreatic cancer.

In one embodiment, the cancer is a glioma.

In one embodiment, the cancer is a glioblastoma.

In one embodiment, the cancer is melanoma.

In one embodiment, the cancer is prostate cancer.

In one embodiment, the cancer is leukemia.

In one embodiment, the cancer is a lymphoma.

In one embodiment, the cancer is non-Hodgkin's lymphoma.

In one embodiment, the cancer is stomach cancer.

In one embodiment, the cancer is lung cancer.

In one embodiment, the cancer is non-small cell lung cancer.

In one embodiment, the cancer is hepatocellular cancer.

In one embodiment, the cancer is gastric cancer.

In one embodiment, the cancer is a stromal tumor of the gastrointestinal tract (GIST).

In one embodiment, the cancer is thyroid cancer.

In one embodiment, the cancer is bile duct cancer.

In one embodiment, the cancer is endometrial cancer.

In one embodiment, the cancer is kidney cancer.

- 7 042928

In one embodiment, the cancer is anaplastic large cell lymphoma.

In one embodiment, the cancer is acute myeloid leukemia (AML).

In one embodiment, the cancer is multiple myeloma.

In one embodiment, the cancer is melanoma.

In one embodiment, the cancer is mesothelioma.

The anti-cancer treatment described above may be used as the sole therapy, or it may include, in addition to the compound of the invention, conventional surgery or radiotherapy or chemotherapy or immunotherapy. Such chemotherapy may be administered concurrently, simultaneously, sequentially, or separately with treatment with a compound of the invention, and may include one or more agents from the following categories of anticancer agents:

(1) antiproliferative/antineoplastic drugs and combinations thereof used in medical oncology, such as alkylating agents (eg, cisplatin, oxaliplatin, carboplatin, cyclophosphamide, chlormethine, melphalan, chlorambucil, busulfan, temozolomide, and nitrosoureas); antimetabolites (eg, gemcitabine and antifolates such as fluoropyrimidines such as 5-fluorouracil and tegafur, raltitrexed, methotrexate, cytosine arabinoside, and hydroxyurea); antitumor antibiotics (eg, anthracyclines such as adriamycin, bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C, dactinomycin, and mithramycin); antimitotic agents (eg vinca alkaloids such as vincristine, vinblastine, vindesine and vinorelbine and taxoids such as taxol and taxotere and polokinase inhibitors); and topoisomerase inhibitors (eg epipodophyllotoxins such as etoposide and teniposide, amsacrine, topotecan and camptothecin);

(2) cytostatic agents such as antiestrogen (eg, tamoxifen, fulvestrant, toremifene, raloxifene, droloxifene, and iodoxifene), antiandrogens (eg, bicalutamide, flutamide, nilutamide, and cyproterone acetate), luteinizing hormone-releasing hormone (LHRH) antagonists, or agonists LHRH (eg goserelin, leuprorelin and buserelin), progestogens (eg megestrol acetate), aromatase inhibitors (eg anastrozole, letrozole, vorazole and exemestane) and 5a-reductase inhibitors such as finasteride;

(3) anti-invasive agents (eg, inhibitors of the c-Src family of kinases such as 4-(6-chloro-2,3methylenedioxyanilino)-7-[2-(4-methylpiperazin-1-yl)ethoxy]-5-tetrahydropyran -4-yloxyquinazoline [AZD0530 (saracatinib); WO01/94341], N(2-chloro-6-methylphenyl)-2-{6-[4-(2-hydroxyethyl)piperazin-1yl]-2-methylpyrimidin-4- ylamino}thiazole-5-carboxamide (dasatinib, BMS-354825; J. Med. Chem., 2004, 47, 6658-6661) and bosutinib (SKI-606), and marimastat-type metalloproteinase inhibitors, inhibitors of plasminogen activator urokinase receptor function, or antibodies to heparanase);

(4) inhibitors of growth factor function: for example, such inhibitors include antibodies to growth factors and antibodies to growth factor receptors (eg, anti-erbB2 antibody trastuzumab [Herceptin™], anti-EGFR antibody panitumumab, anti-erbB1 antibody cetuximab [Erbitux, C225] and any antibodies to growth factors and growth factor receptors disclosed by Stern et al Critical reviews in oncology/haematology, 2005, Vol 54, pp 11-29); such inhibitors also include tyrosine kinase inhibitors, for example epidermal growth factor family inhibitors (for example, EGFR family tyrosine kinase inhibitors such as N-(3-chloro-4-fluorophenyl)-7-methoxy-6-(3morpholinopropoxy)quinazoline-4-amine ( gefitinib, ZD1839), N(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazoline-4-amine (erlotinib, OSI-774) and 6-acrylamido-M-(3-chloro-4-fluorophenyl) -7-(3morpholinopropoxy)quinazoline-4-amine (CI 1033), erbB2 tyrosine kinase inhibitors such as lapatinib); hepatocyte growth factor family inhibitors; inhibitors of the insulin family of growth factors; platelet growth factor family inhibitors such as imatinib and/or nilotinib (AMN107); serine/threonine kinase inhibitors (eg Ras/Raf signaling inhibitors such as farnesyl transferase inhibitors eg sorafenib (BAY 43-9006), tipifarnib (R115777) and lonafarnib (SCH66336)), MEK- and/or AKT- signaling inhibitors kinases, c-kit inhibitors, abl kinase inhibitors, PI3 kinase inhibitors, Plt3 kinase inhibitors, CSF-1R kinase inhibitors, IGF receptor (insulin-like growth factor) kinase inhibitors; aurora kinase inhibitors (eg AZD1152, PH739358, VX-680, MLN8054, R763, MP235, MP529, VX-528 and AX39459) and cyclin-dependent kinase inhibitors such as CDK2 and/or CDK4 inhibitors;

(5) antiangiogenic agents, such as agents that inhibit the effects of vascular endothelial growth factor [eg, the anti-vascular endothelial growth factor antibody bevacizumab [Avastin™] and, for example, a VEGF receptor tyrosine kinase inhibitor, such as vandetanib (ZD6474), vatalanib ( PTK787), sunitinib (SU11248), axitinib (AG-013736), pazopanib (GW 786034), and 4-(4-fluoro-2-methylindol-5-yloxy)-6-methoxy-7-(3-pyrrolidin-1- ilpropoxy)quinazoline (AZD2171;

Example 240 in WO 00/47212), compounds such as those disclosed in WO 97/22596, WO 97/30035, WO 97/32856 and WO 98/13354), and compounds with other mechanisms of action (for example , linomide, inhibitors of αΎβ3 integrin function and angiostatin);

(6) vasoconstrictor agents such as Combretastatin A4 and compounds disclosed in public

- 8 042928 citations of international applications WO 99/02166, WO 00/40529, WO 00/41669, WO 01/92224, WO 02/04434 and

WO 02/08213;

(7) an endothelin receptor antagonist such as zibotentan (ZD4054) or atrasentan;

(8) antisense therapies, such as those directed at the targets listed above, such as ISIS 2503, antisense anti-ras;

(9) gene therapy approaches, including, for example, approaches to replace aberrant genes such as aberrant p53 or aberrant BRCA1 or BRCA2, GDEPT (Gene Targeted Enzyme Prodrug Therapy) approaches such as approaches using cytosine deaminase, thymidine kinase, or bacterial the enzyme nitroreductase, and approaches to increase patient tolerance to chemotherapy or radiotherapy, such as multidrug resistance gene therapy; and (10) immunotherapeutic approaches, including, for example, ex-vivo and in-vivo approaches to increase the immunogenicity of a patient's tumor cells, such as transfection with cytokines such as interleukin 2, interleukin 4, or granulocyte-macrophage colony-stimulating factor, approaches to reduce anergy T -cells, approaches using transfected immune cells, such as cytokine-transfected dendritic cells, approaches using cytokine-transfected tumor cell lines, approaches using anti-idiotypic antibodies, approaches to reduce the function of immunosuppressive cells, such as regulatory T cells, myeloid suppressor cells or IDO (indolamine-2,3,-deoxygenase)-expressing dendritic cells, and approaches using cancer vaccines consisting of proteins or peptides derived from tumor-associated antigens, such as NY-ESO-1, MAGE-3, WT1 or Her2/neu.

Thus, in a further aspect of the invention, there is provided a pharmaceutical product comprising a compound of formula (I) as defined above and an additional antitumor agent as defined above, for the co-treatment of cancer.

In such an aspect of the invention, there is provided a pharmaceutical product comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, as defined above, and an additional antitumor agent, as defined above, for the co-treatment of cancer.

In this description, when the term joint treatment is used with reference to combined treatment, it should be understood that this may refer to simultaneous, separate or sequential administration. References to co-administration should be interpreted in a similar way. In one aspect of the invention, co-treatment refers to simultaneous administration. In another aspect of the invention, co-treatment refers to separate administration. In yet another aspect of the invention, co-treatment refers to sequential administration. When the administration is sequential or separate, the dwell time before the administration of the second component should not be such that the advantage of the effect resulting from the application of the combination is lost. Thus, in one embodiment, sequential administration comprises administering each component of the combination over a period of 11 days. In another embodiment, this period is 10 days. In yet another embodiment, this period is 9 days. In yet another embodiment, this period is 8 days. In yet another embodiment, this period is 7 days. In yet another embodiment, this period is 6 days. In yet another embodiment, this period is 5 days. In yet another embodiment, this period is 4 days. In yet another embodiment, this period is 3 days. In yet another embodiment, this period is 2 days. In another embodiment, this period is 24 hours. In another embodiment, this period is 12 hours.

Thus, in one embodiment, the invention provides the use of a compound of formula (I), or a pharmaceutically acceptable salt thereof, as defined herein, and an additional anticancer agent for the co-treatment of cancer.

In one embodiment, the invention provides the use of a compound of formula (I), or a pharmaceutically acceptable salt thereof, as defined herein, and an additional anticancer agent for the simultaneous, separate, or sequential treatment of cancer.

In one embodiment, a method is provided for producing an anti-cancer effect in a warm-blooded animal, such as a human, in need of such treatment, comprising administering to said animal a compound of formula (I) or a pharmaceutically acceptable salt thereof, and co-administering an additional antitumor agent to said animal, wherein the amounts of compound of formula ( I) or a pharmaceutically acceptable salt thereof and an additional antitumor agent are collectively effective in producing an anticancer effect.

In one embodiment, a method is provided for producing an anti-cancer effect in a warm-blooded animal, such as a human, in need of such treatment, comprising administering to said animal a compound of formula (I) or a pharmaceutically acceptable salt thereof, and simultaneously, separately or sequentially administering an additional antitumor agent to said animal, wherein amounts of a compound of formula (I) or a pharmaceutically acceptable salt thereof and an additional antitumor agent are collectively effective in producing an anticancer effect.

In the manufacture of medicinal compositions, it is important for the drug substance to be in the form in which it can be conveniently processed and processed. This is important not only from the point of view of carrying out an industrially acceptable method of production, but also from the point of view of the subsequent production of pharmaceutical preparations (for example, oral dosage forms such as tablets) containing the active compound.

The different physical properties of the crystalline forms relative to each other and relative to the non-crystalline state can have a significant impact on the chemical and pharmaceutical processing of a compound, especially when the compound is prepared or used on an industrial scale.

In addition, in the manufacture of oral drug compositions, it is important that a reliable and reproducible plasma concentration profile of the drug is achieved after administration to the patient. Variability in the absorption profile of a drug in different patients in the stomach, intestines, or bloodstream can affect the safety and efficacy of the drug.

Chemical stability, solid state stability and shelf life of active ingredients are also very important factors. The drug substance and compositions containing it must be able to maintain efficacy for acceptable periods of time without significantly changing the physico-chemical characteristics of the active component (eg, its chemical composition, density, hygroscopicity and solubility).

Moreover, it is important to be able to offer a drug in a form that is as chemically pure as possible.

Amorphous materials may present problems in this regard. For example, such materials tend to be difficult to process and manufacture, exhibit unreliable solubility, and are often found to be unstable and chemically contaminated.

The person skilled in the art will understand that if the drug can be easily obtained in a stable crystalline form, the above problems can be solved.

Thus, in the manufacture of industrially useful and pharmaceutically acceptable drug compositions, it is important to provide, where possible, the drug in crystalline and stable form.

It should be noted, however, that this goal is not always achievable. Indeed, it is generally not possible to predict, from molecular structure alone, what the crystallization behavior of a compound, either by itself or in salt form, will be. This can only be determined empirically.

In a further aspect of the invention, certain compounds and their salts may be obtained in crystalline forms. These crystalline forms can be characterized as specific polymorphs. When the present invention is said to be in crystalline form, the degree of crystallinity is preferably greater than about 60%, more preferably greater than about 80%, preferably greater than about 90%, and more preferably greater than about 95%. Most preferably, the degree of crystallinity is greater than about 98%.

The specific solid forms described herein exhibit powder X-ray diffraction patterns substantially the same as the X-ray powder diffraction patterns shown in the figures and have individual 2-theta values as shown in the tables included herein. It should be understood that the values of the 2-theta x-ray powder diffraction pattern may vary slightly depending on the equipment or depending on the sample, and such recorded values should not be interpreted as absolute.

It is known that an X-ray powder diffraction pattern having one or more measurement errors can be obtained depending on the measurement conditions (such as the equipment or apparatus used). For example, it is generally known that intensities in an x-ray powder diffraction pattern may deviate depending on measurement conditions. Thus, it should be understood that the solid forms of the present invention are not limited to crystals that give powder X-ray diffraction patterns identical to the X-ray powder diffraction patterns shown in the figures, and any crystals that give powder X-ray diffraction patterns substantially the same as those shown in the figures are within the scope of the present invention. One skilled in the art of X-ray powder diffraction is able to appreciate the similarity in substance of X-ray powder diffraction patterns.

Those skilled in the art of X-ray powder diffraction understand that the relative intensity of the peaks can be affected by, for example, grain sizes above 30 µm, as well as non-unitary relationships, which can affect the analysis of samples. Those skilled in the art also recognize that the position of the reflections can be affected by the exact height at which the sample is placed in the diffractometer, as well as the zero calibration of the diffractometer. The flatness of the sample surface may also have a small effect. Therefore, the reported diffraction pattern data should not be taken as absolute values (Jenkins, R. & Snyder, R.L. 'Introduction to X-Ray Powder Diffractometry' John Wiley & Sons 1996; Bunn, C.W. (1948), Chemical Crystallography, Clarendon Press, London; Klug, H.P. & Alexander, L.E. (1974), X-Ray Diffraction Procedures).

- 10 042928

In general, the diffraction angle measurement error in an X-ray powder diffraction pattern is approximately plus or minus 0.2° 2-theta, and this degree of measurement error should be taken into account when considering the X-ray powder diffraction patterns in the figures, as well as when reading the data, contained in the tables included in this description. Moreover, it should be understood that intensities may vary depending on experimental conditions and sample preparation (preferred orientation).

In this description, K-{2-{2-dimethylaminoethylmethylamino}-4-methoxy-5-{[4-(1-methylindol-3yl)pyrimidin-2-yl]amino}phenyl)prop-2-enamide is designated Compound X The initially formed compound X was found to be an amorphous solid. Several useful crystalline polymorphs have been obtained as a result using the conditions described here in the experimental section. In all embodiments relating to solid forms described here, the peaks in the x-ray powder diffraction patterns are measured using CuKa radiation.

Polymorphic form A of compound X.

Thus, in a further aspect of the invention, a polymorph form A of compound X is provided. This polymorph can be characterized by giving at least one of the following 2θ values measured using CuKa radiation: 7.8 and 21.8.

Polymorph form A of compound X has an X-ray powder diffraction pattern essentially as shown in FIG. 1.

The ten X-ray powder diffraction peaks for this polymorph [2theta angle (2θ), intensity (%)] are 7.8 (100%), 21.8 (73.4%), 13.3 (59.4 %), 6.6 (49.5%), 23.9 (40.5%), 9.6 (38.1%), 14.5 (35.3%), 15.6 (33.2%) %), 22.7 (31.2%) and 19.1 (29.8%).

The present invention provides a Form A polymorph of compound X which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of about 7.8°.

The present invention provides a Form A polymorph of compound X which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of about 21.8°.

The present invention provides a Form A polymorph of compound X which has an X-ray powder diffraction pattern with at least two characteristic peaks at 2-theta values of approximately 7.8° and 21.8°.

The present invention provides a polymorph form A of compound X which has an X-ray powder diffraction pattern with characteristic peaks at 2theta values of about 7.8, 21.8, 13.3, 6.6, 23.9, 9.6, 14.5, 15.6, 22.7 and 19.1°.

The present invention provides a Form A polymorph of compound X which has an X-ray powder diffraction pattern substantially the same as the X-ray powder diffraction pattern shown in FIG. 1.

The present invention provides a Form A polymorph of compound X which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of 7.8° plus or minus 0.2° 2-theta.

The present invention provides a Form A polymorph of compound X which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of 21.8° plus or minus 0.2° 2-theta.

The present invention provides a polymorph form A of compound X which has an x-ray powder diffraction pattern with at least two characteristic peaks at 2-theta values of 7.8° and 21.8°, where said values may differ by plus or minus 0.2° 2-theta.

The present invention provides a polymorph form A of compound X which has an x-ray powder diffraction pattern with characteristic peaks at 2theta values of 7.8, 21.8, 13.3, 6.6, 23.9, 9.6, 14 .5, 15.6, 22.7 and 19.1°, where the indicated values may differ by plus or minus 0.2° 2-theta.

Polymorphic form B of compound X.

In a further aspect of the invention, polymorph form B of compound X is provided. This polymorph can be characterized by giving at least one of the following 2θ values measured using CuKa radiation: 9.3 and 23.4.

Polymorph Form B of compound X has an X-ray powder diffraction pattern essentially as shown in FIG. 3.

The ten X-ray powder diffraction peaks for this polymorph [2theta angle (2θ), intensity (%)] are 9.3 (100%), 23.4 (75.0%), 10.5 (63.6 %), 17.7 (54.3%), 21.0 (48.1%), 16.1 (46.4%), 26.1 (44.2%), 18.6 (41.8%) %), 26.7 (32.2%) and 20.6 (30.9%).

The present invention provides a Form B polymorph of compound X which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of about 9.3°.

The present invention provides a Form B polymorph of compound X which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of approximately 23.4°.

The present invention provides a Form B polymorph of compound X which has an X-ray powder diffraction pattern with at least two characteristic peaks at 2-theta values of approximately 9.3° and 23.4°.

The present invention provides a Form B polymorph of compound X which has an X-ray powder diffraction pattern with characteristic peaks at 2theta values of about 9.3, 23.4, 10.5, 17.7, 21.0, 16.1, 26.1, 18.6, 26.7 and 20.6°.

The present invention provides a Form B polymorph of compound X which has an X-ray powder diffraction pattern substantially the same as the X-ray powder diffraction pattern shown in FIG. 3.

The present invention provides a Form B polymorph of compound X which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of 9.3° plus or minus 0.2° 2-theta.

The present invention provides a Form B polymorph of compound X which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of 23.4° plus or minus 0.2° 2-theta.

The present invention provides a Form B polymorph of compound X which has an X-ray powder diffraction pattern with at least two characteristic peaks at 2-theta values of 9.3° and 23.4°, where said values may differ by plus or minus 0.2° 2-theta.

The present invention provides a Form B polymorph of compound X which has an X-ray powder diffraction pattern with characteristic peaks at 2theta values of 9.3, 23.4, 10.5, 17.7, 21.0, 16.1, 26 ,1, 18.6, 26.7 and 20.6°, where the indicated values may differ by plus or minus 0.2° 2-theta.

Polymorphic form C of compound X.

In a further aspect of the invention, a polymorph form C of compound X is provided. This polymorph can be characterized by giving at least one of the following 2θ values measured using CuKa radiation: 6.0 and 11.3.

Polymorph form C of compound X has an X-ray powder diffraction pattern essentially as shown in FIG. 5.

The ten X-ray powder diffraction peaks for this polymorph [2theta angle (2θ), intensity (%)] are 6.0 (100%), 11.3 (58.2%), 7.5 (40.5 %), 10.3 (21.9%), 12.0 (20.1%), 24.9 (19.4%), 13.0 (16.9%), 14.5 (13.5 %), 16.5 (13.5%) and 18.3 (11.8%).

The present invention provides a Form C polymorph of compound X which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of about 6.0°.

The present invention provides a Form C polymorph of compound X which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of about 11.3°.

The present invention provides a Form C polymorph of compound X which has an X-ray powder diffraction pattern with at least two characteristic peaks at 2-theta values of approximately 6.0° and 11.3°.

The present invention provides a Form C polymorph of compound X which has an X-ray powder diffraction pattern with characteristic peaks at 2theta values of about 6.0, 11.3, 7.5, 10.3, 12.0, 24.9, 13.0, 14.5, 16.5 and 18.3°.

The present invention provides a Form C polymorph of compound X which has an X-ray powder diffraction pattern substantially the same as the X-ray powder diffraction pattern shown in FIG. 5.

The present invention provides a Form C polymorph of compound X which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of 6.0° plus or minus 0.2° 2-theta.

The present invention provides a Form C polymorph of compound X which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of 11.3° plus or minus 0.2° 2-theta.

The present invention provides a Form C polymorph of compound X which has an X-ray powder diffraction pattern with at least two characteristic

- 12 042928 peaks at 2-theta values of 6.0° and 11.3°, where said values may differ by plus or minus 0.2° 2-theta.

The present invention provides a Form C polymorph of compound X which has an X-ray powder diffraction pattern with characteristic peaks at 2theta values of 6.0, 11.3, 7.5, 10.3, 12.0, 24.9, 13 .0, 14.5, 16.5 and 18.3°, where the indicated values may differ by plus or minus 0.2° 2-theta.

Polymorphic form D of compound X.

In a further aspect of the invention, a Form D polymorph of compound X is provided, which is believed to be the monohydrate crystalline form. This polymorph can be characterized by giving at least one of the following 2θ values measured using CuKa radiation: 9.3 and 10.5.

The Form D polymorph of compound X has an X-ray powder diffraction pattern essentially as shown in FIG. 7.

The ten X-ray powder diffraction peaks for this polymorph [2theta angle (2θ), intensity (%)] are 9.3 (100%), 10.5 (90.6%), 16.1 (75.8 %), 26.1 (75.2%), 21.0 (70.9%), 20.6 (56.9%), 16.8 (56.5%), 17.7 (53.3 %), 14.7 (41.3%) and 9.7 (38.3%).

The present invention provides a Form D polymorph of compound X which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of about 9.3°.

The present invention provides a Form D polymorph of compound X which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of about 10.5°.

The present invention provides a Form D polymorph of compound X which has an X-ray powder diffraction pattern with at least two characteristic peaks at 2-theta values of about 9.3° and 10.5°.

The present invention provides a Form D polymorph of compound X which has an X-ray powder diffraction pattern with characteristic peaks at 2theta values of about 9.3, 10.5, 16.1, 26.1, 21.0, 20.6, 16.8, 17.7, 14.7 and 9.7°.

The present invention provides a Form D polymorph of compound X which has an X-ray powder diffraction pattern substantially the same as the X-ray powder diffraction pattern shown in FIG. 7.

The present invention provides a Form D polymorph of compound X which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of 9.3° plus or minus 0.2° 2-theta.

The present invention provides a Form D polymorph of compound X which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of 10.5° plus or minus 0.2° 2-theta.

The present invention provides a Form D polymorph of compound X which has an X-ray powder diffraction pattern with at least two characteristic peaks at 2-theta values of 9.3° and 10.5°, where these values may differ by plus or minus 0.2° 2-theta.

The present invention provides a Form D polymorph of compound X which has an X-ray powder diffraction pattern with characteristic peaks at 2theta values of 9.3, 10.5, 16.1, 26.1, 21.0, 20.6, 16 .8, 17.7, 14.7 and 9.7°, where the indicated values may differ by plus or minus 0.2° 2-theta.

Polymorphic form E of compound X.

In a further aspect of the invention, a polymorph Form E of compound X is provided, which is believed to be the crystalline 1,25-stoichiometric hydrate form of compound X. This polymorph can be characterized by giving at least one of the following 2θ values measured using CuKa radiation: 9.2 and 22.9.

Polymorph form E of compound X has an X-ray powder diffraction pattern essentially as shown in FIG. 10.

The ten X-ray powder diffraction peaks for this polymorph [2theta angle (2θ), intensity (%)] are: 9.2 (100%), 22.9 (84.0%), 14.6 (80, 3%), 12.7 (77.8%), 16.5 (66.4%), 26.9 (60.3%), 9.7 (95.6%), 14.0 (52, 3%), 10.4 (49.9%) and 19.5 (48.3%).

The present invention provides a polymorph Form E of compound X which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of about 9.2°.

The present invention provides a polymorph form E of compound X which has an x-ray powder diffraction pattern with at least one characteristic

- 13 042928 peaking at a 2-theta value of approximately 22.9°.

The present invention provides a polymorph Form E of Compound X which has an X-ray powder diffraction pattern with at least two characteristic peaks at 2-theta values of about 9.2° and 22.9°.

The present invention provides a polymorph Form E of Compound X which has an X-ray powder diffraction pattern with characteristic peaks at 2theta values of about 9.2, 22.9, 14.6, 12.7, 16.5, 26.9, 9.7, 14.0, 10.4 and 19.5°.

The present invention provides a polymorph Form E of Compound X which has an X-ray powder diffraction pattern substantially the same as the X-ray powder diffraction pattern shown in FIG. 10.

The present invention provides a polymorph Form E of compound X which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of 9.2° plus or minus 0.2° 2-theta.

The present invention provides a polymorph Form E of compound X which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of 22.9° plus or minus 0.2° 2-theta.

The present invention provides a polymorph Form E of compound X which has an X-ray powder diffraction pattern with at least two characteristic peaks at 2-theta values of 9.2° and 22.9°, where these values may differ by plus or minus 0.2° 2-theta.

The present invention provides a polymorph Form E of compound X which has an X-ray powder diffraction pattern with characteristic peaks at 2theta values of 9.2, 22.9, 14.6, 12.7, 16.5, 26.9, 9 .7, 14.0, 10.4 and 19.5°, where the indicated values may differ by plus or minus 0.2° 2-theta.

Polymorphic form F of compound X.

In a further aspect of the invention, a polymorph Form F of compound X is provided which is believed to be the 0.25-stoichiometric hydrate form of compound X. This polymorph can be characterized by giving at least one of the following 2θ values measured using CuKa radiation: 18.7 and 8.99.

The F polymorph of compound X has an X-ray powder diffraction pattern essentially as shown in FIG. 13.

The ten X-ray powder diffraction peaks for this polymorph [2theta angle (2θ), intensity (%)] are: 18.7 (100%), 8.9 (87.7%), 15.1 (80, 3%), 25.4 (74.6%), 14.5 (72.3%), 22.9 (69.6%), 9.9 (51.1%), 28.2 (42, 0%), 8.2 (24.2%) and 11.9 (22.3%).

The present invention provides a Form F polymorph of compound X which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of about 18.7°.

The present invention provides a Form F polymorph of compound X which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of about 8.9°.

The present invention provides a Form F polymorph of compound X which has an X-ray powder diffraction pattern with at least two characteristic peaks at 2-theta values of about 18.7° and 8.9°.

The present invention provides a polymorph Form F of compound X which has an X-ray powder diffraction pattern with characteristic peaks at 2theta values of about 18.7, 8.9, 15.1,25.4, 14.5, 22.9, 9.9, 28.2, 8.2 and 11.9°.

The present invention provides a Form F polymorph of compound X which has an X-ray powder diffraction pattern substantially the same as the X-ray powder diffraction pattern shown in FIG. 13.

The present invention provides a Form F polymorph of compound X which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of 18.7° plus or minus 0.2° 2-theta.

The present invention provides a Form F polymorph of compound X which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of 8.9° plus or minus 0.2° 2-theta.

The present invention provides a polymorph Form F of compound X which has an X-ray powder diffraction pattern with at least two characteristic peaks at 2-theta values of 18.7° and 8.9°, where said values may differ by plus or minus 0.2° 2-theta.

The present invention provides a polymorph Form F of compound X which has an X-ray powder diffraction pattern with characteristic peaks at 2theta values of 18.7, 8.9, 15.1.25.4, 14.5.22 .9, 9.9, 28.2, 8.2 and 11.9°, where the indicated values may differ by plus or minus 0.2° 2-theta.

Polymorphic form K of compound X

In a further aspect of the invention, a polymorph form K of compound X is provided. This polymorph can be characterized by giving at least one of the following 2θ values measured using CuKa radiation: 8.4 and 9.7.

The K polymorph of compound X has an X-ray powder diffraction pattern essentially as shown in FIG. 16.

The ten X-ray powder diffraction peaks for this polymorph [2theta angle (2θ), intensity (%)] are: 8.4 (100%), 9.7 (37.7%), 12.2 (32. 4%), 15.1 (25.2%), 24.7 (20.7%), 9.0 (16.8%), 21.9 (13.9%), 19.5 (13, 9%), 24.2 (13.8%) and 18.3 (11.8%).

The present invention provides a Form K polymorph of compound X which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of about 8.4°.

The present invention provides a Form K polymorph of compound X which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of about 9.7°.

The present invention provides a Form K polymorph of compound X which has an X-ray powder diffraction pattern with at least two characteristic peaks at 2-theta values of about 8.4° and 9.7°.

The present invention provides a Form K polymorph of compound X which has an X-ray powder diffraction pattern with characteristic peaks at 2theta values of about 8.4, 9.7, 12.2, 15.1, 24.7, 9.0, 21.9, 19.5, 24.2 and 18.3°.

The present invention provides a Form K polymorph of compound X which has an X-ray powder diffraction pattern substantially the same as the X-ray powder diffraction pattern shown in FIG. 16.

The present invention provides a Form K polymorph of compound X which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of 8.4° plus or minus 0.2° 2-theta.

The present invention provides a Form K polymorph of compound X which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of 9.7° plus or minus 0.2° 2-theta.

The present invention provides a Form K polymorph of compound X which has an X-ray powder diffraction pattern with at least two characteristic peaks at 2-theta values of 8.4° and 9.7°, where said values may differ by plus or minus 0.2° 2-theta.

The present invention provides a polymorph form K of compound X which has an X-ray powder diffraction pattern with characteristic peaks at 2theta values of 8.4, 9.7, 12.2, 15.1, 24.7, 9.0, 21 .9, 19.5, 24.2 and 18.3°, where the indicated values may differ by plus or minus 0.2° 2-theta.

In this description, the mesylate salt of K-(2-{2-dimethylaminoethylmethylamino}-4-methoxy-5-{[4-(1methylindol-3-yl)pyrimidin-2-yl]amino}phenyl)prop-2-enamide is designated as Mesylate salt Y.

Polymorph form A of the mesylate salt Y.

In a further aspect of the invention, polymorph form A of the mesylate salt Y is provided. This polymorph form can be characterized by giving at least one of the following values of 20 measured using CuKa radiation: 5.6 and 6.5.

The polymorph form A of the mesylate salt Y has an X-ray powder diffraction pattern essentially as shown in FIG. 18.

The ten X-ray powder diffraction peaks for this polymorph [2theta angle (2θ), intensity (%)] are: 5.6 (100%), 6.5 (66.7%), 10.2 (97, 2%), 21.0 (96.2%), 13.5 (91.7%), 22.7 (89.6%), 19.3 (80.6%), 27.3 (75, 7%), 15.7 (71.2%) and 19.9 (66.7%).

The present invention provides a polymorph form A of the mesylate salt Y which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of about 5.6°.

The present invention provides a polymorph form A of the mesylate salt Y which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of about 6.5°.

The present invention provides a polymorph form A of the mesylate salt Y which has an x-ray powder diffraction pattern with at least two characteristic peaks at 2-theta values of approximately 5.6° and 6.5°.

The present invention provides a polymorph Form A of the mesylate salt Y which has an X-ray powder diffraction pattern with characteristic peaks at 2-theta values of about 5.6, 6.5, 10.2, 21.0, 13.5, 22 .7, 19.3, 27.3, 15.7 and 19.9°.

The present invention provides a polymorph Form A of the mesylate salt Y which has an X-ray powder diffraction pattern substantially the same as the X-ray powder diffraction pattern shown in FIG. 18.

The present invention provides a polymorph form A of the mesylate salt Y which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of 5.6° plus or minus 0.2° 2-theta.

The present invention provides a polymorph form A of the mesylate salt Y which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of 6.5° plus or minus 0.2° 2-theta.

The present invention provides a polymorph form A of the mesylate salt Y which has an X-ray powder diffraction pattern with at least two characteristic peaks at 2-theta values of 5.6° and 6.5°, where these values may differ by plus or minus minus 0.2° 2-theta.

The present invention provides a polymorph form A of the mesylate salt Y which has an X-ray powder diffraction pattern with characteristic peaks at 2-theta values of 5.6, 6.5, 10.2, 21.0, 13.5, 22, 7, 19.3, 27.3, 15.7 and 19.9°, where the indicated values may differ by plus or minus 0.2° 2-theta.

Polymorphic form B of the mesylate salt of Y.

In a further aspect of the invention, polymorph Form B of the Y mesylate salt is provided. This polymorph can be characterized by giving at least one of the following values of 20 measured using CuKa radiation: 7.2 and 8.6.

Polymorph form B of the mesylate salt Y has an X-ray powder diffraction pattern essentially as shown in FIG. 20.

The ten X-ray powder diffraction peaks for this polymorph [2theta angle (2θ), intensity (%)] are: 7.2 (50.2%), 8.6 (55.2%), 15.3 ( 100%), 10.4 (92.6%), 25.7 (74.0%), 26.1 (63.9%), 16.4 (55.2%), 9.5 (47, 5%), 22.1 (46.9%) and 18.8 (47.7%).

The present invention provides a polymorph Form B of the Y mesylate salt which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of about 7.2°.

The present invention provides a polymorph Form B of the Y mesylate salt which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of about 8.6°.

The present invention provides a polymorph Form B of the Y mesylate salt which has an X-ray powder diffraction pattern with at least two characteristic peaks at 2-theta values of approximately 7.2° and 8.6°.

The present invention provides a polymorph Form B of the Y mesylate salt which has an X-ray powder diffraction pattern with characteristic peaks at 2-theta values of about 7.2, 8.6, 15.3, 10.4, 25.7, 26 .1, 16.4, 9.5, 22.1 and 18.8°.

The present invention provides a polymorph Form B of the Y mesylate salt which has an X-ray powder diffraction pattern substantially the same as the X-ray powder diffraction pattern shown in FIG. 20.

The present invention provides a polymorph Form B of the Y mesylate salt which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of 7.2° plus or minus 0.2° 2-theta.

The present invention provides a polymorph Form B of the Y mesylate salt which has an X-ray powder diffraction pattern with at least one characteristic peak at a 2-theta value of 8.6° plus or minus 0.2° 2-theta.

The present invention provides a polymorph Form B of the Y mesylate salt which has an X-ray powder diffraction pattern with at least two characteristic peaks at 2-theta values of 7.2° and 8.6°, where these values may differ by plus or minus minus 0.2° 2-theta.

The present invention provides a polymorph Form B of the Y mesylate salt which has an X-ray powder diffraction pattern with characteristic peaks at 2-theta values of 7.2, 8.6, 15.3, 10.4, 25.7, 26, 1, 16.4, 9.5, 22.1 and 18.8°, where the indicated values may differ by plus or minus 0.2° 2-theta.

- 16 042928

List of figures

All drawings refer to solid forms of the compound No. Compound X) or its mesylate salt when indicated (Mesylate salt Y).

Fig. 1 is an x-ray powder diffraction pattern of Form A.

Fig. 2 - DSC thermogram form A.

Fig. 3 is an X-ray powder diffraction pattern of Form B.

Fig. 4 - DSC thermogram form B.

Fig. 5 is an X-ray powder diffraction pattern of Form C.

Fig. 6 - DSC thermogram form C.

Fig. 7 is an X-ray powder diffraction pattern of Form D (monohydrate).

Fig. 8 is a DSC thermogram of monohydrate form D.

Fig. 9 is a thermogram of TGA form D monohydrate.

Fig. 10 is an X-ray powder diffraction pattern of Form E (hydrated form).

Fig. 11 - DSC thermogram form E (hydrated form).

Fig. 12 - thermogram of TGA form E (hydrated form).

Fig. 13 is an x-ray powder diffraction pattern of form F (hydrated form).

Fig. 14 is a DSC thermogram of form F (hydrated form).

Fig. 15 - thermogram TGA form F (hydrated form).

Fig. 16 is an x-ray powder diffraction pattern of Form K.

Fig. 17 - DSC thermogram of form K.

Fig. 18 is an x-ray powder diffraction pattern of the mesylate salt Form A.

Fig. 19 is a DSC thermogram of form A of the mesylate salt.

Fig. 20 is an x-ray powder diffraction pattern of mesylate salt Form B.

Fig. 21 - DSC thermogram of form B of the mesylate salt.

Methods of chemical synthesis and biological analysis.

The following abbreviations may be used: THF means tetrahydrofuran; DIPEA means diisopropylethylamine; sat. means saturated aqueous solution; FCC means flash column chromatography using silica; TFA means trifluoroacetic acid; k.t. means room temperature; DMF means N,N-dimethylformamide; DMSO means dimethyl sulfoxide; DMA means N,N-dimethylacetamide; EtOAc means ethyl acetate; h means hour(s); proton NMR: (1H NMR) was determined using deuterated dimethyl sulfoxide at 400 or 500 MHz at about 20-30°C, unless otherwise indicated. Standard abbreviations for NMR are used (s means singlet; d means doublet; dd means doublet of doublets; t means triplet; q means quartet; p means pentet; m means multiplet; br means wide; and so on). When iron is mentioned as a reactant, it is iron powder, 325 mesh and reduced hydrogen. Assay values reported (µM) for this example are concentration values at which 50% inhibition (IC ₅₀ ) is observed. X-ray powder diffraction (XRPD) was performed using Bruker D4 equipment. The X-ray powder diffraction pattern was determined by fixing a sample of crystalline material on a Bruker single crystal silicon wafer (SSC) and flattening the sample into a thin layer using a glass slide. The sample was rotated at a speed of 30 rpm (to improve statistical calculations) and irradiated with X-rays generated by a copper long-fine focus tube operating at 40 kV and 40 mA, with a wavelength of 1.5418 Angstroms (CuKa radiation). A collimated X-ray beam from the source was passed through a setup with automatically controlled slit divergence at V20 (path length 20 mm) and reflected radiation was directed through a 5.89 mm antiscattering slit and a 9.55 mm detector slit. The sample was exposed for 0.03 s in 0.00570° 2-theta steps (continuous scan mode) over a 2-theta angle range of 2 to 40° in theta-theta mode. The run time was 3 min 36 s. This instrument was equipped with a position change detector (Lynxeye). Control and data collection was carried out using a Dell Optiplex 686 NT 4.0 Workstation running the Diffrac+ software product. Differential scanning calorimetry (DSC) was performed using a TA Instruments Q1000 differential scanning calorimeter. Typically, less than 5 mg of material placed in a standard aluminum crucible with a lid was heated over a temperature range of 25°C to 300°C at a constant heating rate of 10°C per minute. A nitrogen gas purge was used at a flow rate of 50 ml per minute. Any crystalline form that produces XRPD patterns or DSC thermograms substantially identical to those described herein is within the scope of the present inventions. One skilled in the art is able to determine the substantive similarity of the diffractograms and the thermograms.

- 17 042928

Assay 1: Exon 19 deletion EGFR cellular phosphorylation assay (activating single mutant).

The human lung cell line PC9 (EGFR with a deletion in exon 19) was obtained from the American Type Culture Collection. PC9 was maintained in RPMI 1640 medium containing 10% fetal calf serum and 2 mM glutamine. Cells were grown in a humidified incubator at 37°C with 5% CO ₂ . Assays to measure cellular phosphorylation of endogenous p-EGFR in cell lysates were performed according to the protocol described in R&D Systems DuoSet IC Human Phospho-EGF RELISA (R&D Systems, catalog number #DYC1095).

μl of cells were seeded (10,000 cells/well) in growth medium in black 384-well clear bottom Corning plates and incubated at 37° C. with 5% CO 2 overnight. Cells were acoustically dosed using an Echo 555 with compounds serially diluted in 100% DMSO. The plates were incubated for an additional 2 hours, then the medium was aspirated and 40 μl of 1x lysis buffer was added to each well. High binding black Greiner 384 well plates were coated with immobilized antibody and then blocked with 3% BSA. After removing the block, 15 µl of lysate was transferred to high binding black 384-well Greiner plates and incubated for 2 h. After aspirating and washing the plates with PBS, 20 µl of detectable antibody was added and incubated for 2 h. µl of fluorogenic peroxidase substrate QuantaBlu (Thermo Fisher Scientific, cat. no. 15169) and incubated for 1 h. radiation 460 nm. The data obtained with each compound was entered into a software package (such as Origin) for analytical curve fitting. Based on these data, IC ₅₀ values were obtained, determined by calculating the concentration of the compound required to obtain a 50% effect.

Assay 2: L858R/T790M EGFR cell phosphorylation assay (double mutant).

Human lung cell lines NCI-H1975 were obtained from the American Type Culture Collection. NCI-H1975 was maintained in RPMI 1640 medium containing 10% fetal bovine serum and 2 mM glutamine. Cells were grown in a humidified incubator at 37°C with 5% CO2. Assays to measure cellular phosphorylation of endogenous p-EGFR in cell lysates were performed according to the protocol described in the R&D Systems DuoSet IC Human Phospho-EGF R ELISA (R&D Systems, catalog number #DYC1095). 40 μl of cells were seeded (10,000 cells/well) in growth medium in black 384-well clear bottom Corning plates and incubated at 37° C. with 5% CO2 overnight. Cells were acoustically dosed using an Echo 555 with compounds serially diluted in 100% DMSO. The plates were incubated for an additional 2 hours, then the medium was aspirated and 40 μl of 1x lysis buffer was added to each well. High binding black Greiner 384 well plates were coated with immobilized antibody and then blocked with 3% BSA. After removing the block, 15 µl of lysate was transferred to high binding black 384-well Greiner plates and incubated for 2 h. After aspirating and washing the plates with PBS, 20 µl of detectable antibody was added and incubated for 2 h. 20 µl of detectable antibody and incubated for 2 h. After aspirating and washing the plates with PBS, 20 µl of QuantaBlu fluorogenic peroxidase substrate (Thermo Fisher Scientific, catalog number 15169) was added and incubated for 1 h. 20 µl of QuantaBlu blocking solution was added to the plates , and fluorescence was read on an Envision plate reader using an excitation wavelength of 352 nm and an emission wavelength of 460 nm. The data obtained with each compound was entered into a software package (such as Origin) for analytical curve fitting. Based on these data, IC ₅₀ values were obtained, determined by calculating the concentration of the compound required to obtain a 50% effect.

Assay 3. Wild type EGFR cellular phosphorylation assay.

The human colon cell line LoVo was obtained from the American Type Culture Collection. LoVo was maintained in RPMI 1640 medium containing 3% depleted fetal calf serum and 2 mM glutamine. Cells were grown in a humidified incubator at 37°C with 5% CO2. Assays to measure cellular phosphorylation of endogenous p-EGFR in cell lysates were performed according to the protocol described in the R&D Systems DuoSet IC Human Phospho-EGF R ELISA (R&D Systems, catalog number #DYC1095). 40 μl of cells were seeded (15,000 cells/well) in growth medium in black 384-well clear bottom Corning plates and incubated at 37° C. with 5% CO2 overnight. Cells were acoustically dosed using an Echo 555 with compounds serially diluted in 100% DMSO. The plates were incubated for an additional 2 hours, then stimulated with 100 ng/ml for 10 minutes and the medium was aspirated and 40 μl of 1x lysis buffer was added to each well. High binding black 384-well Greiner plates were coated with immobilized antibody and then blocked with 3% BSA. After removing the block, 15 µl of lysate was transferred to black 384-well Greiner high binding plates and incubated for 2 h. After aspiration

- 18 042928 and washing the plates with PBS, 20 µl of detectable antibody was added and incubated for 2 h. plates were added with 20 μl of QuantaBlu blocking solution and fluorescence was read on an Envision plate reader using an excitation wavelength of 352 nm and an emission wavelength of 460 nm. The data obtained with each compound was entered into a software package (such as Origin) for analytical curve fitting. Based on these data, IC ₅₀ values were obtained, determined by calculating the concentration of the compound required to obtain a 50% effect.

Analytical data (µM) for the compounds of examples of this application are shown in the table below. Although the data of the analysis are presented with a certain number of significant figures, this should not be taken as a representation that the data are determined to exactly that number of significant figures.

Example No. Analysis 1 Analysis 2 Analysis 3 28 0.01292 0.01144 0.4938 28A 0.01975 0.01271 1.443 29 0.001228 0.0008846 0.04652 thirty 0.07375 0.05211 1.613 31 0.03746 0.00734 2.506 32 0.138 0.02378 10.53 33 0.8916 1.158 11.86 34 0.009044 0.003767 0.1526 35 0.008571 0.006772 0.2623 36 0.04329 0.03272 1.051 37 0.002112 0.001814 0.04859 38 0.005092 0.004405 0.5384 39 0.002336 0.001005 0.2484 40 0.0124 0.01477 >30 41 0.02863 0.0295 1,841 42 0.005192 0.005161 0.4542 43 0.01817 0.01055 1.34 44 0.03329 0.0256 3.64 45 0.1102 0.041 7.396 46 0.1289 0.09293 7.091 47 0.1939 0.1192 15.45 48 0.03988 0.03098 1.579 49 0.0742 0.05097 3.093 50 0.1145 0.1297 7.626 51 0.01296 0.007713 0.4622 52 0.02603 0.01501 1.4 53 0.03537 0.02824 2.638 54 0.003217 0.002803 0.1832 55 0.006433 0.002863 0.5066 56 0.04433 0.02922 3.504 57 0.006455 0.01452 0.08931 58 0.007085 0.01683 0.1786 59 0.002266 0.003021 0.02816 60 0.0146 0.04886 0.6241

Example 28 N-(2-{2-Dimethylaminoethylmethylamino}-4-methoxy-5-{[4-(1-methylindol-3-yl)pyrimidin-2-yl]amino}phenyl)prop-2-enamide

A solution of acryloyl chloride (34.5 mg, 0.38 mmol) in CH ₂ Cl ₂ (1 ml) was added dropwise to a stirred mixture of N ¹ -(2-dimethylaminoethyl)-5-methoxy-N ¹ -methyl-N ⁴ -[ 4-(1-methylindol-3-yl)pyrimidin-2yl]benzene-1,2,4-triamine (intermediate 100, 170 mg, 0.38 mmol) and DIPEA (0.073 ml, 0.42 mmol) in CH ₂ Cl ₂ (5 ml), which was cooled in an ice/water bath. The mixture was stirred for 1.5 h and then diluted with CH ₂ Cl ₂ (25 ml) and washed with sat. NaHCO ₃ (50 ml). The aqueous washes were extracted

- 19 042928

CH ₂ Cl ₂ (2 x 25 ml). The combined organic solutions were dried (MgSO ₄ ) and concentrated in vacuo. Purification by FCC eluting with 0-4% 7N methanolic ammonia in CH ₂ Cl ₂ and trituration with diethyl ether gave the title compound (75 mg, 39%) as an off-white solid; 1Н NMR: 2.21 (6Н, s), 2.29 (2Н, t), 2.72 (3Н, s), 2.89 (2Н, t), 3.86 (3Н, s), 3.92 (3Н, s), 5.77 (1H, dd ), 6.27 (1H, dd), 6.43 (1H, dd), 7.04 (1H, s), 7.15 (1H, t), 7.20-7.27 (2H, m), 7.53 (1H, d), 7.91 (1H, s), 8.24 (1H, d), 8.33 (1H, d), 8.68 (1H, s), 9.14 (1H, s), 10.22 (1H, s); m/z: ES ⁺ MH ⁺ 500.42.

Example 28 (alternative synthesis 1). N(2-{2-Dimethylaminoethylmethylamino}-4-methoxy-5-{[4-(1methylindol-3-yl)pyrimidin-2-yl]amino}phenyl)prop-2-enamide

To a stirred solution of 3-chloro-Ni[2-[2-dimethylaminoethyl(methyl)amino]-4-methoxy-5-[[4-(1methylindol-3-yl)pyrimidin-2-yl]amino]phenyl]propanamide ( intermediate 174, 31.5 g, 58.76 mmol) in acetonitrile (310 ml) at rt. triethylamine (17.84 g, 176.28 mmol) was added. The resulting mixture was heated to 80°C for 6 h, then cooled to rt. Water (130 ml) was then added and the mixture was stirred for 12 h. The mixture was then filtered, washed with a mixture of water and acetonitrile (160 ml, 1:1) and dried at 50° C. overnight to give the title compound (19, 2 g, 94%) as a solid form, identified as polymorph D. 1H NMR: 2.69 (3H, s), 2.83 (6H, d), 3.35 (4H, s), 3.84 (3H, s), 3.91 ( 3Н, s), 5.75 (1H, d), 6.28 (1H, d), 6.67 (1H, dd), 7.05-7.23 (2Н, m), 7.29 (1H, t), 7.43 (1H, d), 7.56 (1H, d), 8.21 (2H, s), 8.81 (1H, s), 9.47 (1H, s), 9.52 (1H, s), m/z: ES ⁺ MH ⁺ 500.26.

Example 28 (alternative synthesis 2). N(2-{2-Dimethylaminoethylmethylamino}-4-methoxy-5-{[4-(1methylindol-3-yl)pyrimidin-2-yl]amino}phenyl)prop-2-enamide

To a stirred solution of N ¹ -(2-dimethylaminoethyl)-5-methoxy-N ¹ -methyl-N ⁴ -[4-(1-methylindol3-yl)pyrimidin-2-yl]benzene-1,2,4-triamine ( intermediate 100, 10 g, 21.32 mmol) in THF (95 ml) and water (9.5 ml) at 0° C. was added 3-chloropropanoyl chloride (3.28 g, 25.59 mmol). The mixture was stirred at rt. for 15 min, then NaOH (3.48 g, 85.28 mmol) was added. The resulting mixture was heated to 65° C. for 10 hours. The mixture was then cooled to rt. and CH ₃ OH (40 ml) and water (70 ml) were added. The resulting mixture was stirred overnight. The resulting solid was collected by filtration, washed with water (25 ml) and dried at 50° C. for 12 h to give the title compound (7.0 g, 94%) as a solid, identified as polymorph D. ¹ H NMR: 2.69 (3Н, s) 2.83 (6Н, d) 3.35 (4Н, s) 3.84 (3Н, s) 3.91 (3Н, s) 5.75 (1H, d) 6.28 (1H, d) 6.67 (1H, dd) 7.05-7.23 (2H, m) 7.29 (1H, t) 7.43 (1H, d) 7.56 (1H, d) 8.21 (2H, s) 8.81 (1H, s) 9.47 (1H, s) 9.52 (1H, s) ES ⁺ MH ⁺ 500.26.

Example 28A. N-(2-{2-Dimethylaminoethylmethylamino}-4-methoxy-5-{[4-(1-methylindol-3-yl)pyrimidin-2-yl]amino}phenyl)prop-2-enamide, mesylate salt

Procedure 1 To a stirred solution of N-[2-[2-dimethylaminoethyl(methyl)amino]-4-methoxy-5[[4-(1-methylindol-3-yl)pyrimidin-2-yl]amino]phenyl]prop -2-enamide (example 28, 20 g, 36.63 mmol) in ethanol (120 ml) and EtOAc (80 ml) at 70° C. methanesulfonic acid (3.59 g, 36.63 mmol) was added as a solution in EtOAc (40 ml). The resulting mixture was stirred for 1.5 hours. The resulting solid was collected by filtration and dried at 80° C. under vacuum overnight to give the title salt (20.5 g, 94%) in a solid form identified as polymorph Form B for this salt.

Procedure 2 To a stirred solution of N-[2-[2-dimethylaminoethyl(methyl)amino]-4-methoxy-5[[4-(1-methylindol-3-yl)pyrimidin-2-yl]amino]phenyl]prop -2-enamide (example 28, 5 g, 9.11 mmol) in acetone (45.5 ml) and water (4.55 ml) at 50° C. methanesulfonic acid (0.893 g, 9.11 mmol) was added as solution in acetone (4.55 ml). The resulting mixture was stirred for 1.5 hours. The resulting solid was collected by filtration and dried at 80° C. under vacuum overnight to give the title salt (4.9 g, 94%) in a solid form defined as polymorph Form B for this salt; ¹ H NMR (acetone-d ⁶ ): 2.72 (3H, s), 2.96 (3H, s), 3.01 (6H, s), 3.58 (3H, t), 3.87-3.90 (7H, m), 5.76 (1H , dd), 6.38-6.53 (2Н, m), 7.12 (1H, t), 7.20 (1H, t), 7.29 (1H, s), 7.40 (2Н, t), 8.07-8.16 (3Н, m), 8.56 (1H, s), 9.30 (1H, s), 9.60 (1H, s), 9.66 (1H, s ); m/z: ES ⁺ MH ⁺ 500.26.

Procedure 3. Polymorph Form A of the Na(2-{2-dimethylaminoethylmethylamino}-4methoxy-5-{[4-(1-methylindol-3-yl)pyrimidin-2-yl]amino}phenyl)prop-2-enamide mesylate salt was prepared in the same manner as described above on a scale of approximately 50 mg, except that acetonitrile was used as the solvent. More specifically, approximately 9.6 mg of methanesulfonic acid was dissolved in a minimum volume of acetonitrile. Approximately 50 mg of N(2-{2-dimethylaminoethylmethylamino}-4-methoxy-5-{[4-(1-methylindol-3-yl)pyrimidin-2-yl]amino}phenyl)prop-2-enamide was also dissolved in the minimum volume of acetonitrile and then the resulting solution was added to a solution of methanesulfonic acid. Upon addition, a solid was formed. This solid was collected by filtration and air dried and then analyzed. The specific solid form obtained in this experiment was designated polymorph form A of this salt.

Intermediate 11 3-(2,5-Dichloropyrimidin-4-yl)-III-indole.

CH ₃ MgBr (3.2 M in 2-methyltetrahydrofuran, 3.37 ml, 10.79 mmol) at 0°C was added dropwise over 10 min to a solution of indole (1.28 g, 10.79 mmol) in THF (6 ml). The solution was then stirred at 02° C. for 0.5 h. Then 2,4,5-trichloropyrimidine (1 g, 5.40 mmol) was added dropwise resulting in a yellow solution. The ice bath was removed, then the solution was stirred at rt. within 1 h, obtaining a red solution. The mixture was heated to 60° C. and then stirred at 60° C. for 1.5 hours. The mixture was then cooled to rt. and acetic acid (634 μl, 11.06 mmol) was added dropwise. Water (9.90 ml) and THF (2 ml) were added, then the mixture was stirred for 20 minutes at 60° C. to give a biphasic solution. The layers were separated and heptane (11 ml) was added to the organic solution resulting in crystallization of the solid. The solid was collected by filtration, washed with heptane (2 ml) and dried in a vacuum oven to give the title compound (1.015 g, 66%) as a yellow solid; 1H NMR: 7.24-7.32 (2H, m), 7.55-7.58 (1H, m), 8.52-8.55 (1H, m), 8.71-8.73 (2H, m), 12.24 (1H, s); m/z: ES ⁺ MH ⁺ 264, 266.

Intermediate 23 4-Fluoro-2-methoxy-5-nitroaniline.

4-Fluoro-2-methoxyaniline (2.4 g, 17.00 mmol) was added portionwise to concentrated H2SO4 (15 mL) which was cooled in an ice/water bath and kept below 15°C during the addition. The mixture was stirred until all the solid that formed had dissolved. KNO3 (0.815 ml, 17.00 mmol) was added in portions so that the temperature was maintained below 10°C. The mixture was stirred overnight and then poured into ice/water. The mixture was made basic with concentrated NH4OH. The resulting solid was filtered and then dissolved in CH ₂ Cl ₂ , washed with water, dried (Na ₂ SO ₄ ) and concentrated on silica. Purification by FCC eluting with 50-0% heptane in CH ₂ Cl ₂ gave the title compound (2.450 g, 77%) as a yellow crystalline solid; ¹ H NMR: 3.91 (3H, s), 5.21 (2H, s), 7.03 (1H, d), 7.35 (1H, d); m/z: ES ⁺ MH ⁺ 187.4.

Intermediate 87 5-Chloro-M-(4-fluoro-2-methoxy-5-nitrophenyl)-4-(1-methylindol-3yl)pyrimidin-2-amine.

Mixture of 3-(2,5-dichloropyrimidin-4-yl)-1-methylindole (intermediate 88, 1281 mg, 4.60 mmol), p-toluenesulfonic acid monohydrate (964 mg, 5.07 mmol) and 4-fluoro -2-methoxy-5-nitroaniline (intermediate 23, 900 mg, 4.84 mmol) in 2-pentanol (50 ml) was heated at 125° C. for 18 hours. A precipitate precipitated from the solution on cooling. The precipitate was collected by filtration, washed with CH3OH (10 ml) and diethyl ether (20 ml) and dried on the filter to give the title compound (1.42 g, 72%) as a tan solid which was used without further purification; ¹ H NMR: 3.91 (3Н, s), 3.96 (3Н, s), 7.05 (1H, t), 7.23-7.3 (1H, m), 7.39 (1H, d), 7.53 (1H, d), 8.33 ( 1H, d), 8.47 (1H, s), 8.58 (1H, s), 8.65 (1H, d), 8.76 (1H, s); m/z: ES ⁺ MH ⁺ 428.10.

Intermediate 88 3-(2,5-Dichloropyrimidin-4-yl)-1-methylindole.

NaH (0.795 g, 19.88 mmol) at 0°C under N2 was added to 3-(2,5-dichloropyrimidin-4-yl)-III-indole (intermediate 11, 5.0 g, 18.9 mmol) in THF (200 mL) and the mixture was stirred at 0°C for 0.25 h. CH3I (1.243 mL, 19.88 mmol) was then added and the mixture was allowed to warm to rt. and stirred for 1 h. The mixture was again cooled in an ice bath and additional NaH (0.795 g, 19.88 mmol) was added. The suspension was stirred at 0°C for 10 min, then CH3I (1.243 ml, 19.88 mmol) was added and the mixture was stirred for 1 h. The mixture was then diluted with water (100 ml), which led to the formation of some solid. The solid was collected by filtration and washed with water and EtOAc and then dried to give the title compound (3.67 g, 70%) as a beige solid. The organic solution was further washed with water and sat. brine and then dried (MgSO4) and concentrated in vacuo. Trituration of the residue with diethyl ether gave a solid which was collected by filtration and dried in vacuo to give the title compound (477 mg, 9%) as a brown solid. This material was only 71% pure, so it was stored separately from the earlier batch;

1H NMR: 3.97 (3H, s), 7.34 (2H, dtd), 7.59-7.65 (1H, m), 8.56 (1H, dd), 8.73 (1H, s), 8.79 (1H, s); m/z: ES+MH+

278.06.

Intermediate 100. N ¹ -(2-Dimethylaminoethyl)-5-methoxy-N ¹ -methyl-N ⁴ -[4-(1methylindol-3-yl)pyrimidin-2-yl]benzene-1,2,4-triamine

Mixture of N-(2-dimethylaminoethyl)-2-methoxy-N-methyl-N-[4-(1-methylindol-3-yl)pyrimidin-2-yl]-5-nitrobenzene-1,4-diamine (intermediate 101, 220 mg, 0.46 mmol), iron (155 mg, 2.78 mmol) and NH ₄ Cl (17.32 mg, 0.32 mmol) in ethanol (12 ml) and water (4 ml) were refluxed for 2 hours. The crude mixture was purified by ion exchange chromatography using an SCX column. The desired product was eluted from the column using 7 M methanolic ammonia and the appropriate fractions were combined and concentrated in vacuo on silica. Purification by FCC eluting with 0-5% 7N methanolic ammonia in CH ₂ Cl ₂ gave the title compound (175 mg, 85%) as a beige foam; 1H NMR: 2.17 (6H, s), 2.36 (2H, t), 2.63 (3H, s), 2.88 (2H, t), 3.74 (3H, s), 3.88 (3H, s), 4.58 (2H, br s), 6.76 (1H, s), 7.12-7.19 (2H, m), 7.21-7.27 (1H, m), 7.48 (1H, s), 7.51 (1H, d), 7.78 (1H, s), 8.27 (1H, d), 8.30 (1H, s), 8.42 (1H, d); m/z: ES ⁺ MH ⁺ 446.32.

Intermediate 101 N'-(2-Dimethylaminoethyl)-2-methoxy-N'-methyl-N-[4-(1methylindol-3-yl)pyrimidin-2-yl]-5-nitrobenzene-1,4-diamine

N ¹ ,N ¹ ,N ² -Trimethylethane-1,2-diamine (80 mg, 0.79 mmol) was added to a suspension of N(4-fluoro-2methoxy-5-nitrophenyl)-4-(1-methylindole-3- yl) pyrimidine-2-amine (Intermediate 129 (which can be prepared as described for Intermediate 87); 350 mg, 0.79 mmol) and DIPEA (0.342 ml, 1.97 mmol) in 2.2.2 -trifluoroethanol (5 ml). The mixture was heated under microwave conditions at 140° C. for 1 hour. The cooled reaction mixture was purified by ion exchange chromatography using an SCX column. The desired product was eluted from the column using 7 M methanolic ammonia and the appropriate fractions were combined and concentrated in vacuo on silica. Purification by FCC eluting with 0-4% 7n. methanolic ammonia in CH ₂ Cl ₂ gave the title compound (230 mg, 62%) as an orange solid; 1H NMR: 2.16 (6H, s), 2.45-2.49 (2H, t, masked by DMSO peak), 2.86 (3H, s), 3.26 (2H, t), 3.87 (3H, s), 3.95 (3H, s) , 6.85 (1H, s), 7.11 (1H, t), 7.21 (1H, d), 7.25 (1H, t), 7.52 (1H, d), 8.10 (1H, s), 8.31 (1H, d), 8.33 (1H, s), 8.36 (1H, d), 8.62 (1H, s); m/z: ES ⁺ MH ⁺ 476.40.

Intermediate 129 N-(4-Fluoro-2-methoxy-5-nitrophenyl)-4-(1-methylindol-3yl)pyrimidin-2-amine

p-Toluenesulfonic acid hydrate (22.73 g, 119.5 mmol) was added in one portion to a mixture of 3(2-chloropyrimidin-4-yl)-1-methylindole (Intermediate 130, 24.27 g, 99.58 mmol) and 4-fluoro2-methoxy-5-nitroaniline (intermediate 23, 18.54 g, 99.58 mmol) in 2-pentanol (500 mL). The resulting mixture was stirred at 105° C. for 2.5 hours and then cooled to rt. The resulting precipitate was collected by filtration, washed with 2-pentanol (50 ml) and dried under vacuum to give some of the desired product as a yellow solid. The filtrate was cooled and the resulting precipitate was collected by filtration and washed with 2-pentanol (10 ml). Two batches of product were combined and triturated with CH3CN to give a solid which was collected by filtration and dried under vacuum to give the title compound (37.4 g, 95%) as a yellow solid; ^1H NMR: 3.92 (3Н, s), 4.01 (3Н, s), 7.13 (1H, dd), 7.27-7.36 (1H, m), 7.40-7.51 (2Н, m), 7.59 (1H, d), 8.26 (1H, t), 8.35 (1H, d), 8.61 (1H, s), 8.85 (1H, d), 9.46 (1H, s); m/z: ES ^- M-392.

Intermediate 130 3-(2-Chloropyrimidin-4-yl)-1-methylindole.

NaH (1.707 g, 42.68 mmol, 40% dispersion in mineral oil) was added in small portions to a chilled (0°C) mixture of 3-(2-chloropyrimidin-4-yl)-1H-indole (intermediate 131,

- 22 042928

8.168 g, 35.57 mmol) in THF (250 ml). The resulting mixture was stirred at 0°C for 0.5 h and then CH ₃ I (2.67 ml, 42.68 mmol) was added and the mixture was stirred at 0°C for 3 additional hours. The reaction mixture was quenched by adding sat. NaHCO ₃ (25 ml). The mixture was then diluted with EtOAc (100 ml) and the resulting solution was washed with sat. NaHCO ₃ (50 ml), water (50 ml) and sat. brine (50 ml). The organic solution was then concentrated in vacuo. Purification by FCC eluting with 0-20% CH ₃ OH in CH ₂ Cl ₂ gave the title compound (8.35 g, 96%) as a pale yellow solid; 1H NMR: 3.90 (3Н, s), 7.30 (2Н, pd), 7.54-7.60 (1H, m), 7.82 (1H, d), 8.38-8.44 (1H, m), 8.49 (1H, s), 8.53 (1H, d); m/z: ES+MH+ 244.

Intermediate 130 3-(2-Chloropyrimidin-4-yl)-1-methylindole (alternative synthesis).

AlCl ₃ (197 g, 1.477 mol) was added portionwise to a solution of 2,4-dichloropyrimidine (200 g, 1342 mmol) in dimethoxyethane (2 L), keeping the temperature below 30°C, and the mixture was stirred for 10 min. Then 1-methylindole (0.172 L, 1.342 mol) was added and the mixture was heated to 80° C. for 2 hours and then left to cool overnight. The mixture was then poured into stirred water (20 L) and then stirred for 1 additional hour. The mixture was then filtered and the resulting solid was washed with water (3 L). The solid was then air dried for 16 hours to give a pink solid (315 g). This solid was then stirred in CH ₃ CN (6.3 L, at reflux) for 1.5 h after which time water (630 ml) was added. The mixture was then allowed to cool to rt. and stirred for 18 hours. Then the mixture was stirred at 5°C for 0.5 hours, then the resulting solid was collected by filtration. The solid was then washed with cold 10% CH ₃ CN/water (2 x 1 L) and then dried to give the title compound (220 g, 67%) as an off-white solid.

Intermediate 131 3-(2-Chloropyrimidin-4-yl)-III-indole.

CH ₃ MgBr (3 M in diethyl ether, 22.68 ml, 68.03 mmol) was added dropwise over a period of 10 min to a stirred solution of SH-indole (7.97 g, 68.03 mmol) in 1.2 -dichloroethane (250 ml) at 0° C. under N ₂ . The resulting solution was stirred for 15 minutes and then 2,4-dichloropyrimidine (15.00 g, 100.69 mmol) was added in one portion. The resulting solution was allowed to warm up to rt. and stirred for 16 additional hours. The reaction mixture was quenched by adding CH ₃ OH (25 ml), then the mixture was concentrated in vacuo and absorbed onto silica. Purification by FCC eluting with 0-20% CH ₃ OH in CH ₂ Cl ₂ gave the title compound (7.17 g, 46%) as a yellow solid; 1H NMR: 7.20-7.28 (2H, m), 7.49-7.53 (1H, m), 7.91 (1H, d), 8.42 (1H, dd), 8.50 (1H, d), 8.53 (1H, d), 12.06 (1H, s); m/z: ES+MH+ 230.

Intermediate 174 3-Chloro-N-[2-[2-dimethylaminoethyl(methyl)amino]-4-methoxy-5-[[4(1-methylindol-3-yl)pyrimidin-2-yl]amino]phenyl ]propanamide

Cl

.o o

To a stirred suspension of N ¹ -(2-dimethylaminoethyl)-5-methoxy-N ¹ -methyl-N ⁴ -[4-(1-methylindol3-yl)pyrimidin-2-yl]benzene-1,2,4-triamine ( intermediate 175, 33 g, 62.29 mmol) and K2CO3 (6.09 g, 43.6 mmol) in acetone (300 ml) 3-chloropropanoyl chloride (9.78 g, 74.74 mmol) was added at -50° WITH. The resulting mixture was heated to -20°C and stirred for 0.5 h. CH3OH (27.75 ml) and NaOH solution (2.24 g, 56.06 mmol in 300 ml water) were added. The resulting mixture was stirred for 3-4 h at rt. The solid was collected by filtration and dried at 50° C. to give the title compound (32.5 g, 95%). ¹ H NMR: (CDCl3) 2.95 (2H, t), 3.04 (6H, d), 3.50 (3H, s), 3.63 (2H, s), 3.81 (2H, t), 4.01 (6H, s), 4.33 -4.37 (2H, m), 7.33-7.42 (3H, m), 7.47 (1H, t), 7.51-7.55 (1H, m), 8.118.21 (3H, m), 8.48 (1H, s), 8.87 (1H, s), 9.17 (1H, s); m/z: ES ⁺ MH ⁺ 536.24.

Intermediate 175. N ⁴ -(2-Dimethylaminoethyl)-2-methoxy-N ⁴ -methyl-N ¹ -[4-(1methylindol-3-yl)pyrimidin-2-yl]-5-nitro-benzene-1, 4-diamine

To a stirred solution of N-(4-fluoro-2-methoxy-5-nitrophenyl)-4-(1-methylindol-3-yl)pyrimidin-2-amine (intermediate 176, 65 g, 160.28 mmol) and N ,N','N'-trimethylethane-1,2-diamine (19.65 g, 192.3 mmol) in DMA (630 ml) was added N-ethyl-N-isopropylpropan-2-amine (26.93 g, 208.4 mmol) at r.t. The resulting mixture was stirred at 85°C for 5-6 h, then cooled to rt. Water (630 ml) was then added and the mixture was stirred for 3-4 hours. The solid material was collected

- 23 042928 by filtration, washed with water (315 ml) and dried at 50°C for 12 h to obtain the title compound (79.4 g, 96%) as an orange solid; 1H NMR (CDCl ₃ ): 2.29 (6H, s),

2.60 (2Н, t), 2.93 (3Н, s), 3.31 (2Н, t), 3.96 (3Н, s), 4.00 (3Н, s), 6.69 (1H, s), 7.21 (1H, d), 7.30 -7.38 (2H, m),

7.43 (1H, d), 7.56 (1H, s), 8.18 (1H, d), 8.30 (1H, s), 8.41 (1H, d), 9.59 (1H, s); m/z: ES+MH+ 476.23.

Intermediate 176 N-(4-Fluoro-2-methoxy-5-nitrophenyl)-4-(1-methylindol-3yl)pyrimidin-2-amine

.O

1,4-Dioxane (585 mL) was added to a mixture of 3-(2-chloropyrimidin-4-yl)-1-methyl-indole (Intermediate 177, 50 g, 160.04 mmol), 4-fluoro-2-methoxy -5-nitroaniline (38.03 g, 192.04 mmol) and p-toluenesulfonic acid monohydrate (37.09 g, 192.04 mmol) at r.t. The resulting mixture was stirred at 85° C. for 3 hours. After cooling to rt. the mixture was quenched with 23% aqueous ammonia (39.59 ml, 480.1 mmol) and water (195 ml, 510.1 mmol) and a solid precipitated.

The resulting suspension was stirred at rt. over 3-4 hours. The solid was collected by filtration and dried at 50° C. under vacuum for 12 hours to give the title compound (74.6 g, 85%) as a yellow solid; ¹ H NMR (CDCl ₃ ): 4.01 (6H, s), 6.90 (1H, d), 7.37-7.48 (4H, m), 8.05-8.12 (2H, m), 8.43 (1H, s), 8.90 (1H , s), 9.34 (1H, s); m/z: ES ⁺ MH ⁺ 394.12.

Intermediate 177 3-(2-Chloropyrimidin-4-yl)-1-methylindole.

To a stirred solution of 2,4-dichloropyrimidine (70.5 g, 463.76 mmol) in dimethoxyethane (900 ml) was added FeCl ₃ (77.16 g, 459.12 mmol) and 1-methylindole (68.28 g) at 60°C. The resulting mixture was stirred overnight at 60°C. After cooling, a solid precipitated by adding methanol (345 ml) and water (900 ml). The resulting suspension was stirred for 3 hours. The solid was collected by filtration, washed with CH ₃ OH (1.38 L) and dried at 50° C. overnight to give the title compound (138.7 g, 81.5%) in as a purple solid; 1H NMR ( _CDCl3 ) 3.89 (3Н, s), 7.36-7.41 (3Н, m), 7.49 (1H, s), 7.96 (1H, s), 8.34 (1H, s), 8.45 (1H, s); m/z: ES ⁺ MH ⁺ 244.05.

Useful crystalline polymorphs of N-(2-{2-dimethylaminoethylmethylamino}-4methoxy-5-{[4-(1-methylindol-3-yl)pyrimidin-2-yl]amino}phenyl)prop-2-enamide (referred to in herein as Compound X) and its mesylate salt (referred to herein as Mesylate salt Y).

Polymorphic form A of compound X.

The initially formed compound X was found to be an amorphous solid. Then, taking some of this amorphous compound X (about 20 mg) and suspending it in cyclohexane (about 2 ml) at 50°C and stirring using a magnetic stirrer for about 4 days, the crystalline polymorph form A of compound X was obtained. Then the sample was allowed to cool, the cap was removed from the vial, and the sample was allowed to dry at ambient conditions to give Form A of Compound X. The X-ray powder diffraction pattern of Form A of Compound X is depicted in FIG. 1. The DSC thermogram of form A of compound X is shown in FIG. 2, which shows an initiating event beginning at 35.1°C and peaking at 50.1°C followed by a melting endotherm beginning at 80.2°C and peaking at 88.3°C.

Polymorphic form B of compound X.

The initially formed compound X was found to be an amorphous solid. Then, taking some of this amorphous compound X (about 20 mg) and dissolving it in the minimum amount of EtOAc required to achieve complete dissolution, the crystalline polymorph form B of compound X was obtained. This solution was then left to evaporate to dryness under ambient conditions to obtain form B of the compound X. The X-ray powder diffraction pattern of Form B of Compound X is shown in FIG. 3. The DSC thermogram of form B of compound X is shown in FIG. 4, which shows a melting endotherm starting at 94.1°C and peaking at 113.6°C.

Polymorphic form C of compound X.

The initially formed compound X was found to be an amorphous solid. Then, taking some of this amorphous compound X (about 20 mg) and dissolving it in the minimum amount of diethyl ether required to achieve complete dissolution, the crystalline polymorph form C of compound X was obtained. This solution was then left to evaporate to dryness under ambient conditions to obtain the form C of Compound X. The X-ray powder diffraction pattern of Form C of Compound X is depicted in FIG. 5. The DSC thermogram of form C of compound X is shown in FIG. 6, which shows a melting endotherm starting at 91.1°C and peaking at 103.8°C.

- 24 042928

Polymorphic form D of compound X.

Form D polymorph of compound X, believed to be the crystalline monohydrate form of compound X, was prepared as described above for Example 28, Alternative Syntheses 1 and 2. The X-ray powder diffraction pattern of Form D of compound X is depicted in FIG. 7. The DSC thermogram of form C of compound X is shown in FIG. 8, which shows a melting endotherm starting at 108.8°C and peaking at 117.7°C. Thermogravimetric analysis showed a weight loss of approximately 3.3%, which is indicative of the monohydrate form (theoretical monohydrate means 3.5%). The TGA thermogram is shown in Fig. 9.

Polymorphic form E of compound X.

Polymorph Form E of Compound X, believed to be the 1.25-stoichiometric hydrate form of Compound X, was prepared by suspending Compound X [154 g, prepared as described for Example 28 (using acryloyl chloride)] in a mixture of methanol (150 ml) and water (600 ml). 10 g of compound X (Form D) was added and the suspension was stirred at rt. within 4 days. The resulting solid was then collected by filtration and washed with water and left to dry. The X-ray powder diffraction pattern of Form E of Compound X is shown in FIG. 10. The DSC thermogram of form E of compound X is depicted in FIG. 11, which shows an initiating event beginning at 66.1°C and peaking at 77.2°C followed by an additional event beginning at 93.6°C and peaking at 101.5°C followed by a melting endotherm beginning at 130.9°C and peak at 135.3°C. Thermogravimetric analysis showed a weight loss of approximately 4.7%, which indicates that the hydrate form is equivalent to 1.25 stoichiometric hydrate (theoretical 1.25 hydrate means 4.3%). The TGA thermogram is shown in Fig. 12.

Polymorphic form F of compound X.

The F polymorph of compound X, which was believed to be the 0.25 stoichiometric hydrate form of compound X, was prepared by drying some form E of compound X in a vacuum oven at rt. to constant weight. The X-ray powder diffraction pattern of Form F of Compound X is shown in FIG. 13. The DSC thermogram of form F of compound X is shown in FIG. 14, which shows an initiating event beginning at 80.9°C and peaking at 92.8°C followed by a melting endotherm beginning at 130.7°C and peaking at 135.7°C. Thermogravimetric analysis showed a weight loss of approximately 0.7%, which indicates that the partially hydrated form is equivalent to a 0.25-stoichiometric hydrate (theoretical 0.25 hydrate means 0.89%). The TGA thermogram is shown in Fig. 15.

Polymorphic form K of compound X.

This polymorphic form of compound X was obtained by the following method.

A solution of acryloyl chloride (0.026 L, 318.48 mmol) in CH ₂ Cl ₂ (290 ml) was added dropwise over 25 min to a stirred suspension of N ¹ -(2-(dimethylamino)ethyl)-5-methoxy-N ¹ -methyl -N ⁴ -(4-(1-methyl-1Hindol-3-yl)pyrimidin-2-yl)benzene-1,2,4-triamine (129 g, 289.52 mmol) in CH ₂ Cl ₂ (2, 9 l), which was cooled to -5°C. Addition is exothermic, but the mixture during the addition was not allowed to heat up more than 1°C. The resulting mixture was stirred at -5°C for 2 hours Then cold feast was added dropwise. NaHCO ₃ solution (1 L), keeping the temperature below -2°C. The mixture was then allowed to warm to rt. The phases were separated and the resulting organic solution was washed with water (100 ml) and saturated brine (100 ml). The solution was then dried (MgSO ₄ ) and concentrated in vacuo. The residue was dissolved in 5% CH3OH in CH ₂ Cl ₂ (60 ml) and then filtered. The filtered solution was purified by FCC eluting with 5% CH3OH in CH ₂ Cl ₂ and the pure fractions were combined and concentrated to give impure compound X as a brown gum (96 g). Further purification by chiral preparative HPLC gave a sample of compound X, which was suspended in CH3OH (50 ml). Not all compound X was able to dissolve. Then water (250 ml) was added and the resulting mixture was suspended overnight with stirring using a magnetic stirrer. The resulting solid was then collected by filtration and dried in a vacuum oven over the weekend to give 16.2 g of compound X in the polymorphic form defined herein as form K. 1H NMR: 2.20 (6H, s), 2.28 (2H, m ), 2.71 (3Н, s), 2.88 (2Н, m), 3.85 (3Н, s), 3.90 (3Н, s), 5.76 (1H, d), 6.27 (1H, d), 6.43 (1H, m) , 7.03 (1H, s), 7.15 (1H, m), 7.22 (2H, m), 7.51 (1H, d), 7.87 (1H, s), 8.23 (1H, m), 8.33 (1H, m), 8.68 (1H, s), 9.18 (1H, s), 10.16 (1H, s).

The X-ray powder diffraction pattern of Form K of Compound X is shown in FIG. 16. The DSC thermogram of form K of compound X is depicted in FIG. 17, which shows a melting endotherm beginning at 129.3°C and peaking at 133.4°C.

Polymorph form A of the mesylate salt Y.

The polymorph form A of the mesylate salt Y was prepared in the manner described previously (Example 28A, Procedure 3). The X-ray powder diffraction pattern of Mesylate Salt Y Form A is shown in FIG. 18. The DSC thermogram of form A of the mesylate salt Y is depicted in FIG. 19, which shows an initiating event beginning at 28.1°C and peaking at 62.2°C followed by a melting endotherm beginning at 258.8°C and peaking at 262.0°C.

- 25 042928

Polymorphic form B of the mesylate salt of Y.

Polymorph form B of the mesylate salt Y was prepared in the manner described previously (Example 28A, procedures 1 and 2). The X-ray powder diffraction pattern of the mesylate salt Form B of the Y mesylate salt is shown in FIG. 20. The DSC thermogram of form B of the Y mesylate salt is shown in FIG. 21, which shows a melting endotherm beginning at 245.0°C and peaking at 246.5°C.

Claims (1)

The polymorphic form of the mesylate salt of K-(2-{2-dimethylaminoethylmethylamino}-4-methoxy-5-{[4-(1methylindol-3-yl)pyrimidin-2-yl]amino}phenyl)prop-2-enamide, which has an x-ray powder diffraction pattern with characteristic peaks at 2-theta values of 7.2; 8.6; 9.5; 10.4; 15.3; 16.4; 18.8; 22.1; 25.7 and 26.1°, where indicated values may differ by plus or minus 0.2° 2-theta measured using CuKa radiation.