U.S. patent application number 15/631230 was filed with the patent office on 2018-01-18 for quinazoline inhibitors of activating mutant forms of epidermal growth factor receptor.
The applicant listed for this patent is AstraZeneca AB. Invention is credited to David Yunzhi Li, Jiabing Wang, Zhenfan Yang, Qingbei Zeng, Xiaolin Zhang.
Application Number | 20180016259 15/631230 |
Document ID | / |
Family ID | 50288181 |
Filed Date | 2018-01-18 |
United States Patent
Application |
20180016259 |
Kind Code |
A1 |
Li; David Yunzhi ; et
al. |
January 18, 2018 |
QUINAZOLINE INHIBITORS OF ACTIVATING MUTANT FORMS OF EPIDERMAL
GROWTH FACTOR RECEPTOR
Abstract
The invention relates to compounds of formula (I), or a
pharmaceutically acceptable salt thereof: ##STR00001## Formula (I)
which possess inhibitory activity against activating mutant forms
of EGFR, and are accordingly useful for their anti-cancer activity
and in methods of treatment of the human or animal body. The
invention also relates pharmaceutical compositions containing them
and to their use in the manufacture of medicaments of use in the
production of an anti-cancer effect in a warm-blooded animal such
as man.
Inventors: |
Li; David Yunzhi; (Shanghai,
CN) ; Wang; Jiabing; (Shanghai, CN) ; Yang;
Zhenfan; (Shanghai, CN) ; Zeng; Qingbei;
(Shanghai, CN) ; Zhang; Xiaolin; (Shanghai,
CN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
AstraZeneca AB |
Sodertalje |
|
SE |
|
|
Family ID: |
50288181 |
Appl. No.: |
15/631230 |
Filed: |
June 23, 2017 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
15162816 |
May 24, 2016 |
9718806 |
|
|
15631230 |
|
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|
|
14709900 |
May 12, 2015 |
9375432 |
|
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15162816 |
|
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|
14197476 |
Mar 5, 2014 |
9066979 |
|
|
14709900 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 39/3955 20130101;
A61K 45/06 20130101; A61K 31/519 20130101; A61P 35/00 20180101;
A61P 35/04 20180101; A61K 31/517 20130101; A61K 31/4184 20130101;
A61K 31/4985 20130101; C07D 403/12 20130101; A61P 35/02 20180101;
A61K 31/519 20130101; A61K 2300/00 20130101; A61K 31/4184 20130101;
A61K 2300/00 20130101; A61K 31/4985 20130101; A61K 2300/00
20130101 |
International
Class: |
C07D 403/12 20060101
C07D403/12; A61K 39/395 20060101 A61K039/395; A61K 31/519 20060101
A61K031/519; A61K 31/517 20060101 A61K031/517; A61K 31/4985
20060101 A61K031/4985; A61K 45/06 20060101 A61K045/06; A61K 31/4184
20060101 A61K031/4184 |
Foreign Application Data
Date |
Code |
Application Number |
Mar 6, 2013 |
CN |
PCT/CN2013/072250 |
Claims
1. A succinate salt of a compound of formula (I): ##STR00024##
2. A succinate salt as claimed in claim 1, which is
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate succinate.
3. The succinate salt as claimed in claim 1, which is in
crystalline form.
4. The succinate salt as claimed in claim 3, which is in
crystalline form having an X-ray powder diffraction pattern with at
least three specific peaks at about 2-theta=6.5.degree.,
17.7.degree. and 14.7.degree..
5. The succinate salt as claimed in claim 4, which is in
crystalline form having an X-ray powder diffraction pattern with
specific peaks at about 2-theta=6.5, 17.7, 14.7, 9.2, 26.5, 20.2,
13.1, 27.3, 24.0.degree..
6. The succinate salt as claimed in claim 1, in association with a
pharmaceutically-acceptable diluent or carrier.
7. A method for the inhibition of activating mutant EGFR in a
warm-blooded animal in need of such treatment, which comprises
administering to said animal an effective amount of the succinate
salt of claim 1.
8. The method of claim 7, wherein the warm-blooded animal is
man.
9. A method of treating cancer in a subject in need thereof,
comprising administering a therapeutically effective amount of the
succinate salt of claim 1.
10. The method of claim 9, wherein the cancer is non-small-cell
lung cancer.
11. The method of claim 10, wherein the non-small-cell lung cancer
is metastatic non-small-cell lung cancer.
12. The method of claim 10, wherein the non-small-cell lung cancer
is non-small-cell lung cancer with CNS metastasis.
13. The method of claim 12, wherein the CNS metastasis is brain
metastasis.
14. The method of claim 12, wherein the CNS metastasis is
leptomeningeal metastasis.
15. The method of claim 9, wherein the succinate salt of claim 1 is
provided in combination with an anti-tumour agent selected from:
(i) an anti-CTLA-4 antibody; (ii)
6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-ca-
rboxylic acid (2-hydroxy-ethoxy)-amide or a pharmaceutically
acceptable salt thereof; (iii) an anti-PD-L1 antibody; (iv)
1-[(1S)-1-(imidazo[1,2-a]pyridin-6-yl)ethyl]-6-(1-methyl-1H-pyrazol-4-yl)-
-1H-[1,2,3]triazolo[4,5-b]pyrazine or a pharmaceutically acceptable
salt thereof; (v) an anti-PD-1 antibody; or (vi) an OX40 agonist
antibody.
16. A succinate salt as claimed in claim 1, in combination with an
anti-tumour agent selected from: (i) an anti-CTLA-4 antibody; (ii)
6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-ca-
rboxylic acid (2-hydroxy-ethoxy)-amide or a pharmaceutically
acceptable salt thereof; (iii) an anti-PD-L1 antibody; (iv)
1-[(1S)-1-(imidazo[1,2-a]pyridin-6-yl)ethyl]-6-(1-methyl-1H-pyrazol-4-yl)-
-1H-[1,2,3]triazolo[4,5-b]pyrazine or a pharmaceutically acceptable
salt thereof; (v) an anti-PD-1 antibody; or (vi) an OX40 agonist
antibody.
17. A compound of the following formula: ##STR00025## or a
pharmaceutically acceptable salt thereof.
18. The compound of claim 17 or a pharmaceutically acceptable salt
thereof, in association with a pharmaceutically-acceptable diluent
or carrier.
Description
[0001] This application is a continuation of U.S. patent
application Ser. No. 15/162,816, filed May 24, 2016, which is a
continuation of U.S. patent application Ser. No. 14/709,900, filed
May 12, 2015, now U.S. Pat. No. 9,375,432, which is a continuation
of U.S. patent application Ser. No. 14/197,476, filed Mar. 5, 2014,
now U.S. Pat. No. 9,066,979, which claims the benefit of priority
to International Application No. PCT/CN2013/072250, filed Mar. 6,
2013, the entire contents of each of which are expressly
incorporated by reference in their entirety.
[0002] The present invention relates to certain
4-(substituted-anilino)-6-O-(substituted-piperizine-carbonyl)quinazoline
compounds and pharmaceutically salts thereof which may be useful in
the treatment or prevention of a disease or medical condition
mediated through activating mutant forms of epidermal growth factor
receptor (EGFR), for example the L858R activating mutant and/or the
Exon 19 deletion activating mutants. Such compounds and salts
thereof may be useful in the treatment or prevention of a number of
different cancers. The invention also relates to pharmaceutical
compositions comprising said compounds, or a pharmaceutically salt
thereof, crystalline forms of these compounds, or a
pharmaceutically salt thereof, intermediates useful in the
manufacture of said compounds, or a pharmaceutically salt thereof,
and to methods of treatment of diseases mediated by activating
mutant forms of EGFR using said compounds, or a pharmaceutically
salt thereof.
[0003] EGFR (otherwise known as ErbB1 or HER1) is a transmembrane
protein tyrosine kinase member of the erbB receptor family. Upon
binding of a growth factor ligand such as epidermal growth factor
(EGF), the receptor can homo-dimerise with another EGFR molecule or
hetero-dimerise with another family member such as erbB2 (HER2),
erbB3 (HER3), or erbB4 (HER4).
[0004] Homo- and/or hetero-dimerisation of erbB receptors results
in the phosphorylation of key tyrosine residues in the
intracellular domain and leads to the stimulation of numerous
intracellular signal transduction pathways involved in cell
proliferation and survival. Deregulation of erbB family signalling
promotes proliferation, invasion, metastasis, angiogenesis, and
tumour cell survival and has been described in many human cancers,
including those of the lung, head and neck and breast.
[0005] The erbB family therefore represents a rational target for
anticancer drug development and a number of agents targeting EGFR
or erbB2 are now clinically available, including gefitinib
(IRESSA.TM.), erlotinib (TARCEVA.TM.) and lapatinib (TYKERB.TM.,
TYVERB.TM.). Detailed reviews of erbB receptor signalling and its
involvement in tumourigenesis are provided in New England Journal
of Medicine [2008] Vol. 358; 1160-74 and Biochemical and
Biophysical Research Communications [2004] Vol. 319: 1-11.
[0006] In 2004 it was reported (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 lung cancer (NSCLC). Approximately 90% of
NSCLC associated EGFR mutations consist of two major EGFR mutations
(E746-A750del in Exon 19 and L858R substitution mutation in Exon
21) (Pao et al. Proceedings of the National Academy of Sciences of
the United States of America [2004], Vol. 13: 306-11 and Kosada et
al. Cancer Research [2004] Vol. 64: 8919-23). These activating
mutations, result in an increase in affinity for small molecule
tyrosine kinase inhibitors such as gefitinib and erlotinib and a
decrease in affinity for adenosine triphosphate (ATP) relative to
wild type (WT) EGFR.
[0007] However, adverse effects, such as skin rash and diarrhoea,
which are considered to be related to inhibition of WT EGFR
signalling pathways in normal skin and gut cells, were reported in
>60% NSCLC patients treated with gefitinib or erlotinib (Zhou C
C et al. Journal of Clinical Oncology [2011], Vol. 12: 735-42; Mok
T S et al. New England Journal of Medicine [2009], Vol. 361:
947-57). In addition, both gefitinib and erlotinib showed limited
effects on treating NSCLC patients with brain metastasis, since
neither of them effectively cross the blood-brain-barrier (BBB)
(McKillop D et al. Xenobiotica [2004], Vol. 34: 983-1000; Jackman D
M et al. Journal of Clinical Oncology [2006], Vol. 24: 4517-20
Grommes C et al. Neuro-Oncology [2011], Vol. 13: 1364-9), while
several reports show that lung cancer brain metastasis are emerging
as an unmet medical need (Gavrilovic et al, Journal of
Neurooncology [2005], Vol. 75: 5-14; Barnholtz-Sloan J S et al.
Journal of Clinical Oncology [2004], 22: 2865-72; Schouten L J et
al, Cancer [2002], Vol. 94: 2698-705).
[0008] Leptomeningeal metastases occur when cancer spreads to the
meninges, the layers of tissue that cover the brain and the spinal
cord. Metastases can spread to the meninges through the blood or
they can travel from brain metastases, carried by the cerebrospinal
fluid (CSF) that flows through the meninges. If tumour cells enter
the CSF and survive, they can travel throughout the central nervous
system, causing neurological problems (Le Rhun et al. Surg
Neurollnt. [2013], Vol. 4: S265-88). The incidence of
leptomeningeal metastases is increasing, partly because cancer
patients are living longer, but also because many chemotherapies
and molecular target therapies are unable to reach sufficient
concentrations in the cerebral spinal fluid to kill the tumour
cells. Treatments have traditionally been ineffective and survival
has been measured in weeks. AstraZeneca has investigated sapitinib
(AZD8931), an equipotent inhibitor of EGFR, HER2 and HER3
receptors, for use in breast cancer. To date sapitinib has been
studied in three phase II clinical trials; the first in combination
with paclitaxel versus paclitaxel alone in advanced breast cancer
patients expressing low levels of HER2; the second in combination
with anastrozole versus anastrozole alone in hormone receptor
positive advanced breast cancer; and the third in combination with
paclitaxel versus paclitaxel alone in metastatic, gastric or
gastro-oesophageal junction cancer who progress following first
line therapy and are ineligible for treatment with trastuzumab by
HER2 status. The compound of the present invention is structurally
distinct from sapitinib, and possesses enhanced brain penetration
properties which make it potentially useful in the treatment of
cancers that have metastasised to the central nervous system [CNS],
particularly those that have metastasised to the brain and those
that result in leptomeningeal metastases.
[0009] Currently some irreversible quinazoline EGFR inhibitors,
such as afatinib and dacomitinib, are under clinical development.
Although these compounds showed comparable effects on EGFR
activating mutations in NSCLC patients with gefitinib and
erlotinib, they demonstrated more severe adverse effects, such as
skin rash (>90% skin rash and diarrhoea) (Zhou C C et al.
Journal of Clinical Oncology [2011], Vol. 12: 735-42; Mok T S et
al. New England Journal of Medicine [2009], Vol. 361: 947-57;
Miller V A et al. Lancet Oncology [2012], Vol. 13: 528-38;
Ramalingam S S et al. Journal of Clinical Oncology [2012], Vol. 30:
3337-44). The compounds of the present invention are reversible
inhibitors, and are therefore expected to have less EGFR-related
adverse effects than afatinib and dacomitinib.
[0010] Certain quinazoline compounds have been disclosed, e.g.
"Preparation of quinazoline derivatives for treatment of tumors"
(US 20080177068 A1), "Preparation of quinazoline derivatives for
treatment of tumors" (US 20080167328 A1), "Preparation of
saccharide derivatives of quinazolines as protein tyrosine kinase
inhibitors" (CN 101857618 A), "Preparation of
chlorofluoroanilinomethoxy-N-methylcarbamoylmethylpiperidinyloxyquinazoli-
ne derivatives for use as antitumor agents" (WO 2010061208 A2),
"Preparation of 4-aminoquinazoline derivatives as antineoplastic
agents (CN 101367793 A)", "Preparation of proline quinazoline
derivatives as antiproliferative agents (BR 2006002275 A)",
"Preparation of quinazoline derivatives as protein kinase
inhibitors" (WO 2005097137 A2), "Preparation of quinazoline
derivatives as protein kinase inhibitors" (WO 2005097134 A2),
"Preparation of quinazoline derivatives as EGFR tyrosine kinase
inhibitors" (WO 2005028469 A1), "Preparation of
phenylamino-substituted quinazolines as inhibitors of EGF and
ErbB-2 kinases" (WO 2005028470 A1), "Preparation of quinazoline
derivatives as EGFR tyrosine kinase inhibitors" (WO 2005026156 A1),
"Preparation of piperidyl-quinazoline derivatives as tyrosine
kinase inhibitors for the treatment of tumors" (WO 2005012290 A1),
"Preparation of 4-anilinoquinazolines as antiproliferative agents"
(WO 2003082831 A1), "Preparation of aminoquinazolines as epidermal
growth factor receptor signal transduction inhibitors" (WO
2002018351 A1), "Preparation of quinazolines as aurora 2 kinase
inhibitors" (WO 2001021594 A1), "Quinazolines and other bicyclic
heterocycles, pharmaceutical compositions containing these
compounds as tyrosine kinase inhibitors, and processes for
preparing them" (WO 2000055141 A1), "Preparation of quinazoline
derivatives and their receptor tyrosine kinase inhibitory
properties" (WO 9738994 A1), "Quinazoline derivatives as antitumor
agents" (WO 9730034 A1), "Preparation of haloanilinoquinazolines as
Class I receptor tyrosine kinase inhibitors" (WO 9633980 A1) and
"Quinazoline derivatives useful for treatment of neoplastic
disease" (U.S. Pat. No. 5,457,105).
[0011] The compounds of the invention, or a pharmaceutically
acceptable salt thereof, when compared with other clinically
available EGFR inhibitors, exhibit certain improved properties e.g.
higher BBB penetration (thus making them potentially useful for the
treatment of cancers that have metastasised to the CNS, in
particular brain metastases and leptomeningeal metastases); show
better selectivity between WT EGFR and mutant EGFR (which may
result in less treatment side effects of skin rash and diarrhoea);
whilst maintaining equivalent or improved activity against
activating mutant EGFR (e.g. EGFR L858R activating mutant and/or
the Exon 19 deletion activating mutants). Therefore, such
compounds, or a pharmaceutically acceptable salt thereof, may be
especially useful in the treatment of disease states in which these
activating mutations of EGFR are implicated, for example in the
treatment of cancer.
[0012] Accordingly, the present invention provides a compound of
formula (I):
##STR00002##
or a pharmaceutically acceptable salt thereof.
[0013] The structures of the clinical compounds referred to above
are as follows:
##STR00003## ##STR00004##
[0014] A suitable pharmaceutically acceptable salt of a compound of
the invention is, for example, an acid-addition salt, for example
an inorganic or organic acid, for example hydrochloric,
hydrobromic, sulphuric, phosphoric, citric, L-tartaric, glycolic,
fumaric, succinic or maleic acid, especially hydrochloric,
hydrobromic, sulphuric, phosphoric, citric, L-tartaric, glycolic,
fumaric or maleic acid. A particular pharmaceutically acceptable
salt of a compound of the invention is a hydrochloric acid salt. A
further particular pharmaceutically acceptable salt of a compound
of the invention is a succinic acid salt. The skilled person would
appreciate that additional acid-addition salts, for example but not
limited to those shown in the examples, may also be possible.
[0015] Salts of the compounds of formula (I) may be formed, for
example, by reacting the compound of formula (I) with an amount of
acid in a medium such as one in which the salt precipitates or in
an aqueous medium followed by lyophilization.
[0016] The compounds of formula (I), or a pharmaceutically
acceptable salt thereof, have a chiral centre. It is to be
understood that the invention encompasses all stereoisomers
(enantiomers and diastereoisomers) of the compounds of formula (I),
or a pharmaceutically acceptable salt thereof, that possess
activating mutant EGFR inhibitory activity. The invention further
relates to any and all tautomeric forms of the compounds of formula
(I), or a pharmaceutically acceptable salt thereof, that possess
activating mutant EGFR inhibitory activity. In a further aspect of
the invention there is provided an enantiomer of formula (I), or a
pharmaceutically acceptable salt thereof, substantially free of any
other enantiomers. In a further aspect of the invention there is
provided the (R)-enantiomer of formula (I), or a pharmaceutically
acceptable salt thereof, substantially free of any other
enantiomers. In a further aspect of the invention there is provided
the (S)-enantiomer of formula (I), or a pharmaceutically acceptable
salt thereof, substantially free of any other enantiomers.
[0017] In one embodiment of the invention where the mixture
comprises unequal molar proportions of enantiomers, the mixture may
have an enantiomeric excess selected from >50%, >70%, >90%
and >95%. Particularly the mixture may have an enantiomeric
excess >98%. More particularly the mixture may have an
enantiomeric excess >99%. More particularly the mixture may have
an enantiomeric excess >99.5%.
[0018] It is also to be understood that certain compounds of
formula (I), or a pharmaceutically acceptable salt thereof, can
exist in solvated as well as unsolvated forms such as, for example,
hydrated forms. It is to be understood that the invention
encompasses all such solvated forms which possess activating mutant
EGFR inhibitory activity.
[0019] It is further to be understood that the invention
encompasses all isotopic forms of the compounds described herein.
For example hydrogen includes deuterium and carbon includes
.sup.12C and .sup.13C.
[0020] In another aspect of the invention, particular compounds of
the invention are any one of the Examples or a pharmaceutically
acceptable salt thereof.
[0021] In another aspect of the invention, particular compounds of
the invention are selected from: [0022]
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate; [0023]
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2S)-2,4-dimethylpiperazine-1-carboxylate; and [0024]
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl (.+-.)
2,4-dimethylpiperazine-1-carboxylate; or a pharmaceutically
acceptable salt thereof.
[0025] In another aspect of the invention, particular compounds of
the invention are selected from: [0026]
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate; [0027]
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2S)-2,4-dimethylpiperazine-1-carboxylate; and [0028]
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl (.+-.)
2,4-dimethylpiperazine-1-carboxylate.
[0029] In another aspect of the invention, a particular compound of
the invention is selected from a pharmaceutically acceptable salt
of 4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate.
[0030] In another aspect of the invention, a particular compound of
the invention is selected from
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate hydrochloride.
[0031] In another aspect of the invention, a particular compound of
the invention is selected from
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate succinate.
[0032] In another aspect of the invention, a particular compound of
the invention is selected from
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate.
[0033] In another aspect of the invention, a particular compound of
the invention is selected from a pharmaceutically acceptable salt
of 4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2S)-2,4-dimethylpiperazine-1-carboxylate.
[0034] In another aspect of the invention, a particular compound of
the invention is selected from
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2S)-2,4-dimethylpiperazine-1-carboxylate.
[0035] In another aspect of the invention, a particular compound of
the invention is selected from a pharmaceutically acceptable salt
of 4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(-)-2,4-dimethylpiperazine-1-carboxylate.
[0036] In another aspect of the invention, a particular compound of
the invention is selected from
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(-)-2,4-dimethylpiperazine-1-carboxylate.
[0037] In another aspect of the invention, a particular compound of
the invention is selected from a pharmaceutically acceptable salt
of 4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(+)-2,4-dimethylpiperazine-1-carboxylate.
[0038] In another aspect of the invention, a particular compound of
the invention is selected from
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(+)-2,4-dimethylpiperazine-1-carboxylate.
[0039] In another aspect of the invention, a particular compound of
the invention is selected from a pharmaceutically acceptable salt
of 4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(.+-.) 2,4-dimethylpiperazine-1-carboxylate.
[0040] In another aspect of the invention, a particular compound of
the invention is selected from
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl (1)
2,4-dimethylpiperazine-1-carboxylate.
[0041] Herein where optical rotations of (+) or (-) are quoted,
particularly they are measured at a c10 where c is the
concentration in g/mL, in DMSO at 25.degree. C. It is also to be
understood that certain compounds of the invention, or a
pharmaceutically acceptable salt thereof, may exist in certain
crystalline forms. In particular
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate, has been identified as
having several crystalline forms--particularly Form A, Form E, Form
I and Form J. In addition the hydrochloride salt of
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate may also exist in
crystalline form--particularly mono-HCl salt Form A.sub.1 and the
succinate salt of
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate may also exist in
crystalline form--particularly succinate salt Form A.sub.8. It is
to be understood that the present invention encompasses all such
crystalline forms of the compounds of formula (I), or a
pharmaceutically acceptable salt thereof, which possess activating
mutant EGFR inhibitory activity.
4-(3-Chloro-2-fluorophenyl)aminol-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate in Crystalline Form, Form
A
[0042] Form A is characterised in providing at least one of the
following 2.degree. values measured using CuKa radiation: 23.3 and
14.3.degree.. Form A is characterised in providing an X-ray powder
diffraction pattern, substantially as shown in FIG. 1. Ten X-Ray
powder diffraction peaks are shown in Table A:
TABLE-US-00001 TABLE A Ten X-Ray Powder Diffraction peaks for Form
A Angle 2-Theta (2.theta.) Intensity % 23.3 100.00 14.3 83.70 9.4
78.08 18.6 61.70 16.3 60.41 21.5 39.61 12.4 38.89 26.1 38.18 19.8
35.71 27.4 31.12
[0043] According to the present invention there is provided a
crystalline form, Form A, which has an X-ray powder diffraction
pattern with at least two specific peaks at about
2-theta=23.3.degree. and 14.3.degree..
[0044] According to the present invention there is provided a
crystalline form, Form A, which has an X-ray powder diffraction
pattern with specific peaks at about 2-theta=23.3, 14.3, 9.4, 18.6,
16.3, 21.5, 12.4, 26.1, 19.8, 27.4.degree..
[0045] According to the present invention there is provided
crystalline form, Form A which has an X-ray powder diffraction
pattern substantially the same as the X-ray powder diffraction
pattern shown in FIG. 1.
[0046] According to the present invention there is provided a
crystalline form, Form A, which has an X-ray powder diffraction
pattern with at least two specific peaks at 2-theta=23.3.degree.
and 14.3.degree. wherein said values may be plus or minus
0.2.degree. 2-theta.
[0047] According to the present invention there is provided a
crystalline form, Form A, which has an X-ray powder diffraction
pattern with specific peaks at 2-theta=23.3, 14.3, 9.4, 18.6, 16.3,
21.5, 12.4, 26.1, 19.8, 27.40 wherein said values may be plus or
minus 0.2.degree. 2-theta.
[0048] DSC analysis of Form A shows a melting endotherm with an
onset of 192.4.degree. C. and a peak at 195.8.degree. C. (FIG.
2).
4-(3-Chloro-2-fluorophenyl)aminol-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate in Crystalline Form, Form
E
[0049] Form E is characterised in providing at least one of the
following 2.theta. values measured using CuKa radiation: 7.3 and
13.7.degree.. Form E is characterised in providing an X-ray powder
diffraction pattern, substantially as shown in FIG. 3. Nine X-Ray
powder diffraction peaks are shown in Table B:
TABLE-US-00002 TABLE B Nine X-Ray Powder Diffraction peaks for Form
E Angle 2-Theta (2.theta.) Intensity % 7.3 100.00 13.7 81.83 13.4
74.07 17.6 28.89 5.6 28.02 10.8 19.08 21.7 19.04 26.5 17.10 28.4
13.41
[0050] According to the present invention there is provided a
crystalline form, Form E, which has an X-ray powder diffraction
pattern with at least two specific peaks at about
2-theta=7.3.degree. and 13.7.degree..
[0051] According to the present invention there is provided a
crystalline form, Form E, which has an X-ray powder diffraction
pattern with specific peaks at about 2-theta=7.3, 13.7, 13.4, 17.6,
5.6, 10.8, 21.7, 26.5, 28.4.degree..
[0052] According to the present invention there is provided
crystalline form, Form E which has an X-ray powder diffraction
pattern substantially the same as the X-ray powder diffraction
pattern shown in FIG. 3.
[0053] According to the present invention there is provided a
crystalline form, Form E, which has an X-ray powder diffraction
pattern with at least two specific peaks at 2-theta=7.3.degree. and
13.7.degree. wherein said values may be plus or minus 0.2.degree.
2-theta.
[0054] According to the present invention there is provided a
crystalline form, Form E, which has an X-ray powder diffraction
pattern with specific peaks at 2-theta=7.3, 13.7, 13.4, 17.6, 5.6,
10.8, 21.7, 26.5, 28.4.degree. wherein said values may be plus or
minus 0.2.degree. 2-theta.
[0055] DSC analysis of Form E shows a melting endotherm with an
onset of 194.2.degree. C. and a peak at 196.3.degree. C. (FIG.
4).
4-(3-Chloro-2-fluorophenyl)aminol-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate in Crystalline Form, Form
I
[0056] Form I is characterised in providing at least one of the
following 2.theta. values measured using CuKa radiation: 3.5 and
7.0.degree.. Form I is characterised in providing an X-ray powder
diffraction pattern, substantially as shown in FIG. 5. Ten X-Ray
powder diffraction peaks are shown in Table C:
TABLE-US-00003 TABLE C Ten X-Ray Powder Diffraction peaks for Form
I Angle 2-Theta (2.theta.) Intensity % 3.5 100.00 7.0 41.22 9.5
32.57 6.4 32.54 14.3 25.70 18.0 24.80 16.4 22.12 15.3 10.95 4.7
7.05 21.3 4.54
[0057] According to the present invention there is provided a
crystalline form, Form I, which has an X-ray powder diffraction
pattern with at least three specific peaks at about 2-theta
3.5.degree., 7.0.degree. and 9.5.degree..
[0058] According to the present invention there is provided a
crystalline form, Form I, which has an X-ray powder diffraction
pattern with specific peaks at about 2-theta=3.5, 7.0, 9.5, 6.4,
14.3, 18.0, 16.4, 15.3, 4.7, 21.3.degree..
[0059] According to the present invention there is provided
crystalline form, Form I which has a X-ray powder diffraction
pattern substantially the same as the X-ray powder diffraction
pattern shown in FIG. 5.
[0060] According to the present invention there is provided a
crystalline form, Form I, which has an X-ray powder diffraction
pattern with at least three specific peaks at 2-theta=3.5.degree.,
7.0.degree. and 9.5.degree. wherein said values may be plus or
minus 0.2.degree. 2-theta.
[0061] According to the present invention there is provided a
crystalline form, Form I, which has an X-ray powder diffraction
pattern with specific peaks at 2-theta=3.5, 7.0, 9.5, 6.4, 14.3,
18.0, 16.4, 15.3, 4.7, 21.3.degree. wherein said values may be plus
or minus 0.2.degree. 2-theta.
[0062] DSC analysis of Form I shows a melting endotherm with an
onset of 193.3.degree. C. and a peak at 195.9.degree. C. (FIG.
6).
4-(3-Chloro-2-fluorophenyl)aminol-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate in Crystalline Form, Form
J
[0063] Form J is characterised in providing at least one of the
following 2.theta. values measured using CuKa radiation: 7.8 and
7.0.degree.. Form J is characterised in providing an X-ray powder
diffraction pattern, substantially as shown in FIG. 7. Ten X-Ray
powder diffraction peaks are shown in Table D:
TABLE-US-00004 TABLE D Ten X-Ray Powder Diffraction peaks for Form
J Angle 2-Theta (2.theta.) Intensity % 7.8 100.00 7.0 49.36 4.9
45.57 15.9 27.11 17.7 20.89 3.4 17.30 20.7 16.71 9.8 14.59 13.9
14.11 12.7 10.83
[0064] According to the present invention there is provided a
crystalline form, Form J, which has an X-ray powder diffraction
pattern with at least two specific peaks at about
2-theta=7.8.degree. and 7.0.degree..
[0065] According to the present invention there is provided a
crystalline form, Form J, which has an X-ray powder diffraction
pattern with specific peaks at about 2-theta=7.8, 7.0, 4.9, 15.9,
17.7, 3.4, 20.7, 9.8, 13.9, 12.7.degree..
[0066] According to the present invention there is provided
crystalline form, Form J which has an X-ray powder diffraction
pattern substantially the same as the X-ray powder diffraction
pattern shown in FIG. 7.
[0067] According to the present invention there is provided a
crystalline form, Form J, which has an X-ray powder diffraction
pattern with at least two specific peaks at 2-theta=7.8.degree. and
7.0.degree. wherein said values may be plus or minus 0.2.degree.
2-theta.
[0068] According to the present invention there is provided a
crystalline form, Form J, which has an X-ray powder diffraction
pattern with specific peaks at 2-theta=7.8, 7.0, 4.9, 15.9, 17.7,
3.4, 20.7, 9.8, 13.9, 12.7.degree. wherein said values may be plus
or minus 0.2.degree. 2-theta.
[0069] DSC analysis of Form J shows a melting endotherm with an
onset of 193.3.degree. C. and a peak at 195.8.degree. C. (FIG.
8).
4-(3-Chloro-2-fluorophenyl)aminol-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate Hydrochloride Salt in
Crystalline Form, Mono-HCl Salt Form A.sub.1
[0070] Mono-HCl salt Form A.sub.1 is characterised in providing at
least one of the following 20 values measured using CuKa radiation:
12.3 and 13.9.degree.. Mono-HCl salt Form A.sub.1 is characterised
in providing an X-ray powder diffraction pattern, substantially as
shown in FIG. 9. Nine X-Ray powder diffraction peaks are shown in
Table E:
TABLE-US-00005 TABLE E Nine X-Ray Powder Diffraction peaks for
mono-HCl salt Form A.sub.1 Angle 2-Theta (2.theta.) Intensity %
12.3 100.00 13.9 40.45 9.3 29.34 23.3 26.42 18.7 20.54 16.0 17.94
24.6 10.24 26.8 8.94 28.0 7.90
[0071] According to the present invention there is provided a
crystalline form, mono-HCl salt Form A.sub.1 which has an X-ray
powder diffraction pattern with at least two specific peaks at
about 2-theta=12.3.degree. and 13.9.degree..
[0072] According to the present invention there is provided a
crystalline form, mono-HCl salt Form A.sub.1 which has an X-ray
powder diffraction pattern with specific peaks at about
2-theta=12.3, 13.9, 9.3, 23.3, 18.7, 16.0, 24.6, 26.8,
28.0.degree..
[0073] According to the present invention there is provided
crystalline form, mono-HCl salt Form A.sub.1 which has an X-ray
powder diffraction pattern substantially the same as the X-ray
powder diffraction pattern shown in FIG. 9.
[0074] According to the present invention there is provided a
crystalline form, mono-HCl salt Form A.sub.1 which has an X-ray
powder diffraction pattern with at least two specific peaks at
2-theta=12.3.degree. and 13.9.degree. wherein said values may be
plus or minus 0.2.degree. 2-theta.
[0075] According to the present invention there is provided a
crystalline form, mono-HCl salt Form A.sub.1 which has an X-ray
powder diffraction pattern with specific peaks at 2-theta=12.3,
13.9, 9.3, 23.3, 18.7, 16.0, 24.6, 26.8, 28.0.degree. wherein said
values may be plus or minus 0.2.degree. 2-theta.
[0076] DSC analysis of mono-HCl salt Form A.sub.1 shows a melting
endotherm with an onset of 259.6.degree. C. and a peak at
261.4.degree. C. (FIG. 10).
4-[(3-Chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate Succinate Salt in
Crystalline Form, Succinate Salt Form A.sub.8
[0077] Succinate salt Form A.sub.8 is characterised in providing at
least one of the following 20 values measured using CuKa radiation:
6.5 and 17.7. Succinate salt Form A.sub.8 is characterised in
providing an X-ray powder diffraction pattern, substantially as
shown in FIG. 11. Nine X-Ray powder diffraction peaks are shown in
Table F:
TABLE-US-00006 TABLE F Angle 2-Theta (2.theta.) Intensity % 6.5
100.00 17.7 31.30 14.7 24.91 9.2 21.73 26.5 14.13 20.2 12.03 13.1
11.74 27.3 9.72 24.0 5.56
[0078] According to the present invention there is provided a
crystalline form, Succinate salt Form A.sub.8 which has an X-ray
powder diffraction pattern with at least three specific peaks at
about 2-theta=6.5.degree., 17.7.degree. and 14.7.degree..
[0079] According to the present invention there is provided a
crystalline form, Succinate salt Form A.sub.8 which has an X-ray
powder diffraction pattern with specific peaks at about
2-theta=6.5, 17.7, 14.7, 9.2, 26.5, 20.2, 13.1, 27.3,
24.0.degree..
[0080] According to the present invention there is provided
crystalline form, Succinate salt Form A.sub.8 which has an X-ray
powder diffraction pattern substantially the same as the X-ray
powder diffraction pattern shown in FIG. 11.
[0081] According to the present invention there is provided a
crystalline form, Succinate salt Form A.sub.8 which has an X-ray
powder diffraction pattern with at least three specific peaks at
2-theta=6.5.degree., 17.7.degree. and 14.7.degree. wherein said
values may be plus or minus 0.2.degree. 2-theta.
[0082] According to the present invention there is provided a
crystalline form, Succinate salt Form A.sub.8 which has an X-ray
powder diffraction pattern with specific peaks at 2-theta=6.5,
17.7, 14.7, 9.2, 26.5, 20.2, 13.1, 27.3, 24.0.degree. wherein said
values may be plus or minus 0.2.degree. 2-theta.
[0083] DSC analysis shows Succinate salt Form A.sub.8 shows a
melting endotherm with an onset of 191.8.degree. C. and a peak at
194.2.degree. C. (FIG. 12).
LEGENDS TO FIGURES
[0084] FIG. 1: X-Ray Powder Diffraction Pattern of Form A
[0085] FIG. 2: DSC Thermogram of Form A
[0086] FIG. 3: X-Ray Powder Diffraction Pattern of Form E
[0087] FIG. 4: DSC Thermogram of Form E
[0088] FIG. 5: X-Ray Powder Diffraction Pattern of Form I
[0089] FIG. 6: DSC Thermogram of Form I
[0090] FIG. 7: X-Ray Powder Diffraction Pattern of Form J
[0091] FIG. 8: DSC Thermogram of Form J
[0092] FIG. 9: X-Ray Powder Diffraction Pattern of mono-HCl salt
Form A.sub.1
[0093] FIG. 10: DSC Thermogram of mono-HCl salt Form A.sub.1
[0094] FIG. 11: X-Ray Powder Diffraction Pattern of Succinate salt
Form A.sub.8
[0095] FIG. 12: DSC Thermogram of Succinate salt Form A.sub.8
[0096] When it is stated that the present invention relates to a
crystalline form, the degree of crystallinity is conveniently
greater than about 60%, more conveniently greater than about 80%,
conveniently greater than about 90% and more conveniently greater
than about 95%. Most conveniently the degree of crystallinity is
greater than about 98%.
[0097] It will be understood that the 2-theta values of the X-ray
powder diffraction pattern may vary slightly from one machine to
another or from one sample to another, and so the values quoted are
not to be construed as absolute. It is known that an X-ray powder
diffraction pattern may be obtained which has one or more
measurement errors depending on measurement conditions (such as
equipment or machine used). In particular, it is generally known
that intensities in an X-ray powder diffraction pattern may
fluctuate depending on measurement conditions. Therefore it should
be understood that the polymorphic forms of the present invention
are not limited to the crystals that provide X-ray powder
diffraction patterns identical to the X-ray powder diffraction
pattern shown in the figures, and any crystals providing X-ray
powder diffraction patterns substantially the same as those shown
in the figures fall within the scope of the present invention. A
person skilled in the art of X-ray powder diffraction is able to
judge the substantial identity of X-ray powder diffraction
patterns.
[0098] Persons skilled in the art of X-ray powder diffraction will
realise that the relative intensity of peaks can be affected by,
for example, grains above 30 microns in size and non-unitary aspect
ratios, which may affect analysis of samples. The skilled person
will also realise that the position of reflections can be affected
by the precise height at which the sample sits in the
diffractometer and the zero calibration of the diffractometer. The
surface planarity of the sample may also have a small effect. Hence
the diffraction pattern data presented are not to 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).
[0099] Generally, a measurement error of a diffraction angle in an
X-ray powder diffractogram is approximately plus or minus
0.2.degree. 2-theta, and such degree of a measurement error should
be taken into account when considering the X-ray powder diffraction
patterns shown in the figures and tables. Furthermore, it should be
understood that intensities might fluctuate depending on
experimental conditions and sample preparation (preferred
orientation).
[0100] Therefore, in a further aspect of the invention there is
provided
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate in crystalline form.
[0101] In a further aspect of the invention there is provided a
pharmaceutically acceptable salt of
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate in crystalline form.
[0102] In a further aspect of the invention there is provided a
hydrochloride salt of
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate in crystalline form.
[0103] In one aspect of the invention,
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate in crystalline form is in
the form of Form A.
[0104] In one aspect of the invention,
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate in crystalline form is in
the form of Form E.
[0105] In one aspect of the invention,
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate in crystalline form is in
the form of Form I.
[0106] In one aspect of the invention,
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate in crystalline form is in
the form of Form J.
[0107] In one aspect of the invention, the hydrochloride salt of
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate in crystalline form is in
the form of mono-HCl salt Form A.sub.1.
[0108] In one aspect of the invention, the succinate salt of
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate in crystalline form is in
the form of succinate salt Form A.sub.8.
[0109] In one aspect of the invention,
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate in crystalline form is in
the form of Form A and is substantially free of any other
Forms.
[0110] In one aspect of the invention,
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate in crystalline form is in
the form of Form E and is substantially free of any other
Forms.
[0111] In one aspect of the invention,
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate in crystalline form is in
the form of Form I and is substantially free of any other
Forms.
[0112] In one aspect of the invention,
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate in crystalline form is in
the form of Form J and is substantially free of any other
Forms.
[0113] In one aspect of the invention the hydrochloride salt of
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate in crystalline form is in
the form of mono-HCl salt Form A.sub.1 and is substantially free of
any other Forms.
[0114] In one aspect of the invention the succinate salt of
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate in crystalline form is in
the form of Succinate salt Form A.sub.8 and is substantially free
of any other Forms.
[0115] The term "substantially free" refers to less than 10% of
another Form or Forms, enantiomer or enantiomers, particularly less
than 5%. In another aspect "substantially free" refers to less than
1% of another Form or Forms, enantiomer or enantiomers. Here in
Form also includes the amorphous Form.
[0116] As stated hereinbefore the compounds, or a pharmaceutically
acceptable salt thereof, defined in the present invention possess
anti-cancer activity which is believed to arise from the activating
mutant EGFR inhibitory activity, and other properties, of the
compounds, or a pharmaceutically acceptable salt thereof. These
properties may be assessed, for example, using the procedures set
out below.
Assay 1: Cellular Phosphorylation Assay
[0117] The human lung cell line NCI-H3255 (L858R) was obtained from
the American Type Culture Collection. The NCI-H3255 cells were
maintained in BEBM media (Lonza; CC-3171), containing 10% fetal
bovine serum (FBS) (Gibco; 10099-141), supplemented with BEGM kit
(Lonza; CC-4175). The human lung cell line PC-9 (Exon 19 deletion
EGFR) was obtained from the American Type Culture Collection. PC-9
cells were maintained in RPMI 1640 (Gibco; 22400-089), containing
10% fetal bovine serum. The human lung cell line NCI-H838 (EGFR
wild type) was obtained from the American Type Culture Collection.
NCI-H838 cells were maintained in RPMI 1640 (Gibco; 22400-089),
containing 10% fetal bovine serum.
[0118] All cells were grown in a humidified incubator at 37.degree.
C. with 5% CO.sub.2. Assays to measure cellular phosphorylation of
endogenous p-EGFR in cell lysates were carried out according to the
protocol described in the PathScan.RTM. Phospho-EGF Receptor
(Tyr1068) Sandwich ELISA Kit (Cell Signalling kit catalogue number
#7240).
[0119] 100 .mu.L of cells were seeded (32000 cells/well) in RPMI
1640+1% fetal bovine serum in Corning Costar, 96 well cell culture
plates and incubated at 37.degree. C. with 5% CO.sub.2 overnight.
Cells were acoustically dosed using a Tecan, with compounds
serially diluted in 100% DMSO. Cell plates were incubated for a
further 4 h after the compounds were added, (for NCI-H838: rhEGF
(R&D catalogue number#236-EG) was added to cell plate with
final concentration 100 ng/ml rhEGF to stimulate 5 minutes), then
following aspiration of medium, 110 .mu.L IP lysis buffer (IP lysis
buffer: add 1:100 phosphatase inhibitor cocktail 2&3 (Sigma
catalogue number P5726&P0044), 1:100 protease inhibitor
cocktail (Sigma catalogue number P8340) to Pierce IP lysis buffer
(Thermo catalogue number #87788)) was added to each well. The
plates were put at 4.degree. C. with rotation 300 rpm for 0.5-1
hour. 100 .mu.l/well of cell lysis was transferred to coated plates
(Cell Signalling kit catalogue number#7240) and incubated overnight
at 4.degree. C. with rotation 300 rpm. The plates we taken from
4.degree. C. to 37.degree. C. with rotation 300 rpm for 1 hour.
Following aspiration and washing of the plates with 1.times. wash
buffer, 100 .mu.l of detection antibody (Cell Signalling kit
catalogue number#7240) was added to each well. The plate was sealed
with tape and incubated for 2 hours at 37.degree. C. with rotation
300 rpm. Following aspiration and washing of the plates with
1.times. wash buffer, 100 .mu.l of HRP-linked secondary antibody
(Cell Signalling kit catalogue number#7240) was added to each well.
The plate was sealed with tape and incubated for 1 hour at
37.degree. C. with rotation 300 rpm. Following aspiration and
washing of the plates with 1.times. wash buffer, 100 .mu.l of TMB
substrate (Cell Signalling kit, catalogue number#7240) was added to
each well. The plate was sealed with tape and incubated for 30
minutes at 37.degree. C. with 300 rpm. 100 .mu.l stop solution
(Cell Signalling kit catalogue number#7240) was added to the plates
and absorbance read at 450 nm within 30 minutes on SpectraMax M5e
plate reader.
[0120] The data obtained with each compound was exported into a
suitable software package (such as H-BASE) to perform curve fitting
analysis. From this data an IC.sub.50 value was determined by
calculation of the concentration of compound that is required to
give a 50% effect.
[0121] The assay data (.mu.M) in Assay 1 for the Examples of this
application as well as that obtained for gefitinib and erlotinib
are shown in the table below (where n=the number of times the
experiment was repeated):
TABLE-US-00007 IC.sub.50 IC.sub.50 IC.sub.50 Compound (NCI-H3255)
(PC-9) (NCI-H838) Example 1 0.0072 .+-. 0.0013 (n = 12) 0.0074 .+-.
0.0013 (n = 12) 0.065 .+-. 0.009 (n = 13) Example 3 0.005 (n = 1)
0.008 (n = 1) 0.04 (n = 1) Example 4 0.001 (n = 1) 0.004 (n = 1)
0.04 (n = 1) gefitinib 0.0065 .+-. 0.002 (n = 42) 0.0062 .+-.
0.0019 (n = 42) 0.03 .+-. 0.01 (n = 42) erlotinib 0.0081 .+-.
0.0019 (n = 10) 0.0061 .+-. 0.0019 (n = 10) 0.033 .+-. 0.007 (n =
10)
[0122] This shows that Example 1, Example 2, and Example 3 have
comparable potency to gefitinib and erlotinib.
Assay 2: Brain Blood Barrier Penetration Assay
[0123] Both K.sub.p,uu brain and K.sub.p,uu CSF should be the main
parameters measured and optimized in CNS drug discovery (Di L et
al., Journal of Medicinal Chemistry [2013], 56: 2-12). K.sub.p,uu
brain, the relationship between concentrations of unbound drug in
brain and in blood, predicts drug action on metastatic tumors in
brain Leptomeningeal metastasis (LM) results from metastatic spread
of cancer to the leptomeninges, giving rise to central nervous
system dysfunction. K.sub.p,uu CSF represents the distribution of
drug in CSF as compared to that in blood, which drives drug
response during leptomeningeal metastasis treatment.
[0124] In vitro blood and brain binding assay was carried out on a
HT-Dialysis plate (Gales Ferry, Conn.) with semi-permeable
membrane. Diluted blood (1:1 with DPBS pH7.4) and brain homogenate
(1:3 with DPBS pH7.4) were spiked with 5 .mu.M test compound (in
triplicate) and dialyzed against equal volume of 150 .mu.L 100 mM
PBS buffer (pH7.4) at 37.degree. C. for 4 hours in a slowly rotated
plate. At the end of incubation, a 50 .mu.L aliquot from the
receiver side and a 5 L from the donor chamber were taken. The 5
.mu.L sample was further diluted with 45 .mu.L of blank blood or
brain homogenate. Paired samples were matrix-matched with either
buffer or blank blood/brain homogenate and mixed for 2 min, and
then precipitated with 150 .mu.L cold acetonitrile with 100 ng/mL
tolbutamide as internal standard. After centrifuging at 4000 rpm
for 20 min, supernatant was diluted with 0.1% formic acid aqueous
solution and analyzed for LC/MS/MS (API 4000, Applied Biosystems,
Foster City). Unbound fraction (fu) of test compound in the brain
homogenate and diluted blood were calculated by the ratio of the
buffer side response to the brain homogenate/blood side response,
and unbound fraction (f.sub.u,bl and f.sub.u,br) of test compound
in non-diluted blood and tissue were calculated from measured fu in
homogenate and diluted blood with the following equation:
f.sub.u,bl (f.sub.u,br)=(1/D)/[(1/fu-1)+1/D)]. D is dilution
factor.
[0125] A Short oral absorption (SOA) model is an in-vivo screening
model to identify brain penetration of a compound. Six male Han
Wistar rats purchased from Beijing Vital River were orally dosed
with the compound at 2 mg/kg in 1% methylcellulose. At 0.25, 0.5,
1, 2, 4 and 7 hour post-dose, cerebral spinal fluid (CSF) was
collected from cisterna magna, and blood samples (>60 .mu.L/time
point/each site) were collected via cardiac puncture, into separate
EDTA coagulated tubes, and then immediately diluted with 3-fold
volume of water. Brain tissue was harvested and homogenized in
3.times. volume of 100 mM phosphate buffered saline (pH7.4). All
samples were stored at .about.-70.degree. C. prior to LC/MS/MS
analysis.
[0126] Standards were prepared by spiking blank blood, brain
homogenate and artificial CSF covering 0.2 to 500 ng/mL.
Homogenized brain tissue along with blood samples were precipitated
by adding 3-fold volume of cold acetonitrile containing internal
standard (40 ng/mL Dexamethasone and 40 ng/mL Diclofenac), and 10
.mu.L of CSF samples were precipitated with 100 .mu.L of cold
acetonitrile containing internal standard. After 2 min vortex and 5
min centrifugation at 14,000 rpm, supernatant was analyzed by
LC/MS/MS (API 4000, Applied Biosystems, Foster City). Two sets of
standard curves were run at the beginning and end of each batch
from blood sample analysis. For brain and CSF samples, one standard
curve was analyzed along with test samples.
[0127] Total brain levels, expressed as brain/blood ratio
(Kp.sub.,brain) were measured by AUC.sub.brain/AUC.sub.blood in
rodents after oral administration. Free fraction of test compound
in biological matrix was determined by in vitro blood and brain
binding assay. K.sub.p,uu brain and K.sub.p,uu CSF were calculated
by the following equation: K.sub.p,uu
brain=AUC.sub.brain/AUC.sub.blood.times.(f.sub.u,brain/f.sub.u.blood)
and K.sub.p,uu
CSF=AUC.sub.CSF/(AUC.sub.blood.times.f.sub.u.blood).
[0128] The assay data in Assay 2 for the Examples of this
application as well as data obtained for sapitinib (freebase form)
is shown in the table below:
TABLE-US-00008 Compound K.sub.p, uu brain K.sub.p, uu CSF Example 1
0.8-1.3 (n = 2) 1.0-1.3 (n = 2) Example 3 1.6 (n = 1) 2.6 (n = 1)
sapitinib 0.13 (n = 1) Below quantification limit
demonstrating the superior brain barrier penetration properties of
the compounds of the present invention, when compared to
sapitinib.
[0129] According to a further aspect of the invention there is
provided a pharmaceutical composition which comprises a compound of
formula (I), or a pharmaceutically acceptable salt thereof, as
defined hereinbefore, in association with a
pharmaceutically-acceptable diluent or carrier.
[0130] The composition may be in a form suitable for oral
administration, for example as a tablet or capsule, for parenteral
injection (including intravenous, subcutaneous, intramuscular,
intravascular or infusion) as a sterile solution, suspension or
emulsion, for topical administration as an ointment or cream or for
rectal administration as a suppository. Particularly the
composition may be in a form suitable for oral administration.
[0131] In general the above compositions may be prepared in a
conventional manner using conventional excipients.
[0132] The compound of formula (I), or a pharmaceutically
acceptable salt thereof, will be administered to a warm-blooded
animal at a unit dose within the range 0.01-2000 mg/kg,
particularly 2.5-1000 mg/kg, particularly 5-500 mg/kg, and this
should provide a therapeutically effective dose. However the daily
dose will necessarily be varied depending upon the host treated,
the particular route of administration, and the severity of the
illness being treated. Accordingly the optimum dosage may be
determined by the practitioner who is treating any particular
patient.
[0133] According to a further aspect of the present invention there
is provided a compound of formula (I), or a pharmaceutically
acceptable salt thereof, as defined hereinbefore for use in a
method of treatment of the human or animal body by therapy.
[0134] As a result of its activating mutant EGFR inhibitory
activity, the compounds of formula (I), or a pharmaceutically
acceptable salt thereof, are expected to be useful in the treatment
of diseases or medical conditions mediated alone or in part by
activating mutant EGFR, for example cancer. The types of cancers
which may be susceptible to treatment using the compounds of
formula (I), or a pharmaceutically acceptable salt thereof,
include, but are not limited to, ovarian cancer, cervical cancer,
colorectal cancer, breast cancer, pancreatic cancer, glioma,
glioblastoma, melanoma, prostate cancer, leukaemia, lymphoma,
non-Hodgkins lymphoma, lung cancer, hepatocellular cancer, gastric
cancer, gastrointestinal stromal tumour, thyroid cancer, bile duct
cancer, endometrial cancer, renal cancer, anaplastic large cell
lymphoma, acute myeloid leukaemia, multiple myeloma, melanoma and
mesothelioma. In a particular embodiment of the invention, the type
of cancer which may be susceptible to treatment using the compound
of formula (I), or a pharmaceutically acceptable salt thereof is
non-small-cell lung cancer (NSCLC). In a further particular
embodiment the NCSLC cells in the warm blooded animal possess or
have previously been shown to possess activation mutations in the
EGFR gene.
[0135] The compound of formula (I), or a pharmaceutically
acceptable salt thereof, is useful in the treatment of disease
states in which activating mutant EGFR is implicated. In one aspect
of the invention where activating mutant EGFR is referred to this
refers one or more mutations in the ATP-binding site (kinase
domain) of the EGFR gene, particularly around Exons 18-21, such as
those described in WO 2005/094357. In one aspect of the invention
where activating mutant EGFR is referred to this refers to L858R
activating mutant EGFR and/or Exon 19 deletion activating mutant
EGFR. In one aspect of the invention where activating mutant EGFR
is referred to this refers to L858R activating mutant EGFR and Exon
19 deletion activating mutant EGFR. In one aspect of the invention
where activating mutant EGFR is referred to this refers to L858R
activating mutant EGFR. In another aspect of the invention where
activating mutant EGFR is referred to this refers to Exon 19
deletion activating mutant EGFR.
[0136] It is envisaged that for the methods of treatment of cancer
mentioned herein, the compounds of formula (I), or a
pharmaceutically acceptable salt thereof, will be administered to a
mammal, more particularly a human being. Similarly, the uses of the
compounds of formula (I), or a pharmaceutically acceptable salt
thereof, for the treatment of cancer mentioned herein, it is
envisaged that the compounds of formula (I), or a pharmaceutically
acceptable salt thereof, will be administered to a mammal, more
particularly a human being.
[0137] According to another aspect of the invention, there is
therefore provided the compounds of formula (I), or a
pharmaceutically acceptable salt thereof, as defined hereinbefore
for use as a medicament.
[0138] According to a further aspect of the invention there is
provided the use of a compound of formula (I), or a
pharmaceutically acceptable salt thereof, as defined hereinbefore
in the manufacture of a medicament for the inhibition of activating
mutant EGFR in a warm-blooded animal such as man.
[0139] According to this aspect of the invention there is provided
the use of a compound of formula (I), or a pharmaceutically
acceptable salt thereof, as defined hereinbefore in the manufacture
of a medicament for the production of an anti-cancer effect in a
warm-blooded animal such as man.
[0140] According to a further feature of the invention, there is
provided the use of a compound of formula (I), or a
pharmaceutically acceptable salt thereof, as defined hereinbefore
in the manufacture of a medicament for use in the treatment of
ovarian cancer, cervical cancer, colorectal cancer, breast cancer,
pancreatic cancer, glioma, glioblastoma, melanoma, prostate cancer,
leukaemia, lymphoma, non-Hodgkins lymphoma, lung cancer,
hepatocellular cancer, gastric cancer, gastrointestinal stromal
tumour, thyroid cancer, bile duct cancer, endometrial cancer, renal
cancer, anaplastic large cell lymphoma, acute myeloid leukaemia,
multiple myeloma, melanoma and mesothelioma.
[0141] According to a further feature of the invention, there is
provided the use of a compound of formula (I), or a
pharmaceutically acceptable salt thereof, as defined hereinbefore
in the manufacture of a medicament for use in the treatment of
NSCLC.
[0142] According to a further feature of this aspect of the
invention there is provided a method of inhibiting activating
mutant EGFR in a warm-blooded animal, such as man, in need of such
treatment which comprises administering to said animal an effective
amount of a compound of formula (I), or a pharmaceutically
acceptable salt thereof, as defined hereinbefore.
[0143] According to a further feature of this aspect of the
invention there is provided a method for producing an anti-cancer
effect in a warm-blooded animal, such as man, in need of such
treatment which comprises administering to said animal an effective
amount of a compound of formula (I), or a pharmaceutically
acceptable salt thereof, as defined hereinbefore.
[0144] According to a further feature of this aspect of the
invention there is provided a method for producing an anti-cancer
effect in a warm-blooded animal, such as man, in need of such
treatment which comprises (1) determining whether or not the warm
blooded animal has an activating EGFR mutation in the tumour cell
and (2) and if so administering to said animal an effective amount
of the compound of formula (I), or a pharmaceutically acceptable
salt thereof, as defined hereinbefore.
[0145] According to an additional feature of this aspect of the
invention there is provided a method of treating ovarian cancer,
cervical cancer, colorectal cancer, breast cancer, pancreatic
cancer, glioma, glioblastoma, melanoma, prostate cancer, leukaemia,
lymphoma, non-Hodgkins lymphoma, lung cancer, hepatocellular
cancer, gastric cancer, gastrointestinal stromal tumour, thyroid
cancer, bile duct cancer, endometrial cancer, renal cancer,
anaplastic large cell lymphoma, acute myeloid leukaemia, multiple
myeloma, melanoma and mesothelioma, in a warm-blooded animal, such
as man, in need of such treatment which comprises administering to
said animal an effective amount of a compound of formula (I), or a
pharmaceutically acceptable salt thereof, as defined
hereinbefore.
[0146] According to an additional feature of this aspect of the
invention there is provided a method of treating NSCLC, in a
warm-blooded animal, such as man, in need of such treatment which
comprises administering to said animal an effective amount of a
compound of formula (I), or a pharmaceutically acceptable salt
thereof, as defined hereinbefore.
[0147] According to a further aspect of the invention there is
provided a compound of formula (I), or a pharmaceutically
acceptable salt thereof, as defined hereinbefore for use in
inhibiting activating mutant EGFR in a warm-blooded animal such as
man.
[0148] According to this aspect of the invention there is provided
a compound of formula (I), or a pharmaceutically acceptable salt
thereof, as defined hereinbefore for use in the production of an
anti-cancer effect in a warm-blooded animal such as man.
[0149] According to a further feature of the invention, there is
provided a compound of formula (I), or a pharmaceutically
acceptable salt thereof, as defined hereinbefore for use in the
treatment of ovarian cancer, cervical cancer, colorectal cancer,
breast cancer, pancreatic cancer, glioma, glioblastoma, melanoma,
prostate cancer, leukaemia, lymphoma, non-Hodgkins lymphoma, lung
cancer, hepatocellular cancer, gastric cancer, gastrointestinal
stromal tumour, thyroid cancer, bile duct cancer, endometrial
cancer, renal cancer, anaplastic large cell lymphoma, acute myeloid
leukaemia, multiple myeloma, melanoma and mesothelioma.
[0150] According to a further feature of the invention, there is
provided a compound of formula (I), or a pharmaceutically
acceptable salt thereof, as defined hereinbefore for use in the
treatment of NSCLC.
[0151] In a further aspect of the invention there is provided a
pharmaceutical composition which comprises a compound of formula
(I), or a pharmaceutically acceptable salt thereof, as defined
hereinbefore in association with a pharmaceutically-acceptable
diluent or carrier for use in inhibiting activating mutant EGFR in
a warm-blooded animal such as man.
[0152] In a further aspect of the invention there is provided a
pharmaceutical composition which comprises a compound of formula
(I), or a pharmaceutically acceptable salt thereof, as defined
hereinbefore in association with a pharmaceutically-acceptable
diluent or carrier for use in the production of an anti-cancer
effect in a warm-blooded animal such as man.
[0153] In a further aspect of the invention there is provided a
pharmaceutical composition which comprises a compound of formula
(I), or a pharmaceutically acceptable salt thereof, as defined
hereinbefore in association with a pharmaceutically-acceptable
diluent or carrier for use in the treatment of ovarian cancer,
cervical cancer, colorectal cancer, breast cancer, pancreatic
cancer, glioma, glioblastoma, melanoma, prostate cancer, leukaemia,
lymphoma, non-Hodgkins lymphoma, lung cancer, hepatocellular
cancer, gastric cancer, gastrointestinal stromal tumour, thyroid
cancer, bile duct cancer, endometrial cancer, renal cancer,
anaplastic large cell lymphoma, acute myeloid leukaemia, multiple
myeloma, melanoma and mesothelioma in a warm-blooded animal such as
man.
[0154] In a further aspect of the invention there is provided a
pharmaceutical composition which comprises a compound of formula
(I), or a pharmaceutically acceptable salt thereof, as defined
hereinbefore in association with a pharmaceutically-acceptable
diluent or carrier for use in the treatment of NSCLC in a
warm-blooded animal such as man.
[0155] In any of the aspects or embodiments mentioned herein where
cancer is mentioned said cancer may be selected from ovarian
cancer, cervical cancer, colorectal cancer, breast cancer,
pancreatic cancer, glioma, glioblastoma, melanoma, prostate cancer,
leukaemia, lymphoma, non-Hodgkins lymphoma, lung cancer,
hepatocellular cancer, gastric cancer, gastrointestinal stromal
tumour, thyroid cancer, bile duct cancer, endometrial cancer, renal
cancer, anaplastic large cell lymphoma, acute myeloid leukaemia,
multiple myeloma, melanoma and mesothelioma.
[0156] In any of the aspects or embodiments mentioned herein where
cancer is mentioned, particularly said cancer may be selected from
lung cancer. In a further aspect, particularly said cancer may be
selected from non-small-cell lung cancer. In a further aspect,
particularly said cancer may be selected from non-metastatic
non-small-cell lung cancer. In a further aspect, particularly said
cancer may be selected from metastatic non-small-cell lung
cancer.
[0157] The compound of the present invention may be applied in the
adjuvant and/or 1.sup.st line and/or 2.sup.nd line treatment
settings of NSCLC patients carrying activating mutant EGFR, with
and without CNS metastasis, particularly brain metastasis and/or
leptomeningeal metastases.
[0158] In another aspect the cancer is in a non metastatic
state.
[0159] In another aspect the cancer is in a metastatic state.
[0160] In another aspect of the invention particularly the
metastasis are CNS metastases.
[0161] In another aspect, particularly the CNS metastases are brain
metastases.
[0162] In another aspect, particularly the CNS metastases are
leptomeningeal metastases. Certain NSCLC patients with CNS
metastasis, particularly brain metastasis and/or leptomeningeal
metastases, exhibit CNS symptoms, such as headache and vomiting.
For these patients, whole brain radiation therapy (WBRT) may be
used to improve these symptoms. The compound of the present
invention may be able to enhance the anti-tumour effect of WBRT as
well as to further improve CNS symptoms when used in combination
with WBRT.
[0163] The activating mutant EGFR activity treatment defined
hereinbefore may be applied as a sole therapy or may involve, in
addition to the compound of the invention, conventional surgery or
radiotherapy (for example WBRT as described hereinabove) or
chemotherapy. Such chemotherapy may include one or more of the
following anti-tumour agents:--
[0164] (i) an anti-CTLA-4 antibody;
[0165] (ii)
6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-ca-
rboxylic acid (2-hydroxy-ethoxy)-amide (as disclosed in WO
2007/076245) or a pharmaceutically acceptable salt thereof;
[0166] (iii) an anti-PD-L1 antibody;
[0167] (iv)
1-[(1S)-1-(imidazo[1,2-a]pyridin-6-yl)ethyl]-6-(1-methyl-1H-pyrazol-4-yl)-
-1H-[1,2,3]triazolo[4,5-b]pyrazine (Compound 270 of WO 2011/079804)
or a pharmaceutically acceptable salt thereof;
[0168] (v) an anti-PD-1 antibody; or
[0169] (vi) an OX40 agonist antibody.
[0170] Particularly an anti-CTLA-4 antibody is tremelimumab (as
disclosed in U.S. Pat. No. 6,682,736).
[0171] In another aspect of the invention, particularly an
anti-CTLA-4 antibody is ipilimumab (marketed by Bristol Myers Squib
as YERVOY.RTM.).
[0172] Particularly
"6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidazole-5-c-
arboxylic acid (2-hydroxy-ethoxy)-amide (as disclosed in WO
2007/076245) or a pharmaceutically acceptable salt thereof" is the
hydrogen sulphate salt of
6-(4-bromo-2-chloro-phenylamino)-7-fluoro-3-methyl-3H-benzoimidaz-
ole-5-carboxylic acid (2-hydroxy-ethoxy)-amide. More particularly
the hydrogen sulphate salt is 1:1 compound: H.sub.2SO.sub.4.
[0173] Particularly an anti-PD-L1 antibody is an antibody as
disclosed in US 20130034559 (MedImmune). In another aspect of the
invention particularly an anti-PD-L1 antibody is an antibody as
disclosed US 2010/0203056 (Genentech/Roche). In another aspect of
the invention particularly an anti-PD-L1 antibody is an antibody as
disclosed US 20090055944 (Medarex). In another aspect of the
invention particularly an anti-PD-L1 antibody is an antibody as
disclosed US 20130323249 (Sorrento Therapeutics).
[0174] Particularly an anti-PD-1 antibody is MRK-3475 as disclosed
in WO 2009/114335 and U.S. Pat. No. 8,168,757 (Merck). In another
aspect of the invention particularly is Nivolumab, an anti-PD-1
antibody as disclosed in WO 2006/121168 or U.S. Pat. No. 8,008,449
(Medarex). In another aspect of the invention particularly an
anti-PD-1 antibody is an antibody as disclosed in WO2009/101611
(CureTech). In another aspect of the invention particularly an
anti-PD-1 antibody is an antibody as disclosed in WO2012/145493
(Amplimmune). In another aspect of the invention particularly an
anti-PD-1 antibody is an antibody as disclosed in U.S. Pat. No.
7,488,802 (Wyeth/MedImmune).
[0175] Particularly an anti-OX40 antibody is an antibody as
disclosed in US20110123552 (Crucell). In another aspect of the
invention particularly an anti-PD-1 antibody is an antibody as
disclosed in US 20130280275 (Board of Regents, Univ. of Texas). In
another aspect of the invention particularly an anti-PD-1 antibody
is an antibody as disclosed in WO 99/42585 (Agonox) and WO 95/12673
and WO 95/21915.
[0176] According to this aspect of the invention there is provided
a combination suitable for use in the treatment of cancer
comprising a compound of formula (I) as defined hereinbefore or a
pharmaceutically acceptable salt thereof and any one of the anti
tumour agents listed under (i)-(iv) above.
[0177] Therefore in a further aspect of the invention there is
provided a compound of formula (I) or a pharmaceutically acceptable
salt thereof in combination with an anti-tumour agent selected from
one listed under (i)-(iv) herein above. Herein, where the term
"combination" is used it is to be understood that this refers to
simultaneous, separate or sequential administration. In one aspect
of the invention "combination" refers to simultaneous
administration. In another aspect of the invention "combination"
refers to separate administration. In a further aspect of the
invention "combination" refers to sequential administration. Where
the administration is sequential or separate, the delay in
administering the second component should not be such as to lose
the beneficial effect of the combination.
[0178] According to a further aspect of the invention there is
provided a pharmaceutical composition which comprises a compound of
formula (I) or a pharmaceutically acceptable salt thereof in
combination with an anti-tumour agent selected from one listed
under (i)-(iv) herein above, in association with a pharmaceutically
acceptable diluent or carrier.
[0179] According to a further aspect of the invention there is
provided a pharmaceutical composition which comprises a compound of
formula (I) or a pharmaceutically acceptable salt thereof in
combination with an anti-tumour agent selected from one listed
under (i)-(iv) herein above, in association with a pharmaceutically
acceptable diluent or carrier for use in producing activating
mutant EGFR activity.
[0180] According to a further aspect of the invention there is
provided a pharmaceutical composition which comprises a compound of
formula (I) or a pharmaceutically acceptable salt thereof in
combination with an anti-tumour agent selected from one listed
under (i)-(iv) herein above, in association with a pharmaceutically
acceptable diluent or carrier for use in producing an anti-cancer
effect.
[0181] According to a further aspect of the invention there is
provided a pharmaceutical composition which comprises a compound of
formula (I) or a pharmaceutically acceptable salt thereof in
combination with an anti-tumour agent selected from one listed
under (i)-(iv) herein above, in association with a pharmaceutically
acceptable diluent or carrier for use in treating ovarian cancer,
cervical cancer, colorectal cancer, breast cancer, pancreatic
cancer, glioma, glioblastoma, melanoma, prostate cancer, leukaemia,
lymphoma, non-Hodgkins lymphoma, lung cancer, hepatocellular
cancer, gastric cancer, gastrointestinal stromal tumour, thyroid
cancer, bile duct cancer, endometrial cancer, renal cancer,
anaplastic large cell lymphoma, acute myeloid leukaemia, multiple
myeloma, melanoma and mesothelioma.
[0182] According to a further aspect of the invention there is
provided a pharmaceutical composition which comprises a compound of
formula (I) or a pharmaceutically acceptable salt thereof in
combination with an anti-tumour agent selected from one listed
under (i)-(iv) herein above, in association with a pharmaceutically
acceptable diluent or carrier for use in treating NSCLC.
[0183] According to a further aspect of the present invention there
is provided a kit comprising a compound of formula (I) or a
pharmaceutically acceptable salt thereof in combination with an
anti-tumour agent selected from one listed under (i)-(iv) herein
above.
[0184] According to a further aspect of the present invention there
is provided a kit comprising: [0185] a) a compound of formula (I)
or a pharmaceutically acceptable salt thereof in a first unit
dosage form; [0186] b) an anti-tumour agent selected from one
listed under (i)-(iv) herein above; in a second unit dosage form;
and [0187] c) container means for containing said first and second
dosage forms.
[0188] In addition to their use in therapeutic medicine, the
compounds of formula (I), or a pharmaceutically acceptable salt
thereof, are also useful as pharmacological tools in the
development and standardisation of in vitro and in vivo test
systems for the evaluation of the effects of activating mutant EGFR
inhibitory activity in laboratory animals such as cats, dogs,
rabbits, monkeys, rats and mice, as part of the search for new
therapeutic agents.
[0189] In the above other pharmaceutical composition, process,
method, use and medicament manufacture features, the alternative
and preferred embodiments of the compounds of the invention
described herein also apply.
EXAMPLES
[0190] The invention will now be illustrated in the following
Examples in which, generally:
[0191] (i) in general, the course of reactions were followed by
liquid chromatography mass spectrometry (LCMS) or thin later
chromatography (TLC); the reaction times that are given are not
necessarily the minimum attainable;
[0192] (ii) when necessary, organic solutions were dried over
anhydrous magnesium sulfate or anhydrous sodium sulfate, work-up
procedures were carried out using traditional layer separating
techniques, evaporations were carried out either by rotary
evaporation under reduced pressure or in a Genevac HT-4/EZ-2.
[0193] (iii) yields, where present, are not necessarily the maximum
attainable, and when necessary, reactions were repeated if a larger
amount of the reaction product was required;
[0194] (iv) in general, the structures of the end-products were
confirmed by nuclear magnetic resonance (NMR) and/or mass spectral
techniques; electrospray mass spectral data were obtained using a
Waters ZMD or Waters ZQ LC/mass spectrometer acquiring both
positive and negative ion data, generally, only ions relating to
the parent structure are reported; proton NMR chemical shift values
were measured on the delta scale at 400 MHz using a Bruker NMR
spectrometer or a Varian NMR spectrometer. The following
abbreviations have been used: s, singlet; d, doublet; pd, partial
doublet; t, triplet; q, quartet; m, multiplet; br, broad.
Exchangeable protons are not always observed or reported in the NMR
of end-products due to exchange with deuterated solvent or
advantageous deuterated water in the solvent or the signal is
poorly resolved and/or very broad;
[0195] (v) intermediates were not necessarily fully purified but
their structures and purity were assessed by TLC, analytical HPLC
and/or NMR analysis;
[0196] (vi) unless otherwise stated, column chromatography (by the
flash procedure) and medium pressure liquid chromatography (MPLC)
were performed on Merck Kieselgel silica (Art. 9385) or by using
pre-packed silica cartridges on semi-automated flash chromatography
(SFC) equipment (for example a CombiFlash Companion); and
[0197] (vii) the following abbreviations have been used: [0198] Boc
tert-butyloxycarbonyl; [0199] CD.sub.3OD deuteromethanol; [0200]
DMSO-d.sub.6 hexadeuterodimethylsulfoxide; [0201] CDCl.sub.3
deuterochlorform; [0202] PE petroleum ether; [0203] IPA
isopropanol; [0204] iPrOAc isopropyl acetate; [0205] MTBE methyl
tert-butyl ether; [0206] DCM dichloromethane; [0207] THF
tetrahydrofuran; [0208] RT room temperature; [0209] MeOH methanol;
[0210] EtOH ethanol; and [0211] EtOAc ethyl acetate.
X-Ray Powder Diffraction
[0212] Analytical Instrument: Panalytical Empyrean. The X-ray
powder diffractogram was determined by mounting a sample of the
crystalline material on a Si single crystal holder and spreading
out the sample into a thin layer with the aid of a microscope
slide. The 20 position was calibrated against Panalytical 640 Si
powder standard. The sample irradiated with X-rays generated by a
copper long-fine focus tube operated at 45 kV and 40 mA with a
wavelength of K.alpha.1=1.540598 angstroms and K.alpha.2=1.544426
angstroms (K.alpha.2/K.alpha.1 intensity ratio is 0.50). The
collimated X-ray source was passed through an programmed divergence
slit set at 10 mm and the reflected radiation directed through a
5.5 mm antiscatter slit. The sample was exposed for 12.7 seconds
per 0.0167.degree. 2-theta increment (continuous scan mode) over
the range 3 degrees to 40 degrees 2-theta in theta-theta mode. The
running time was 3 minutes and 57 seconds. The instrument was
equipped with a RTMS detector (X'Celerator). Control and data
capture was by means of a Dell Optiplex 780 XP operating with data
collector software. Persons skilled in the art of X-ray powder
diffraction will realize that the relative intensity of peaks can
be affected by, for example, grains above 30 microns in size and
non-unitary aspect ratios that may affect analysis of samples. The
skilled person will also realize that the position of reflections
can be affected by the precise height at which the sample sits in
the diffractometer and the zero calibration of the diffractometer.
The surface planarity of the sample may also have a small effect.
Hence the diffraction pattern data presented are not to be taken as
absolute values.
Differential Scanning Calorimetry
[0213] Analytical Instrument: TA Instruments Q200 or Q2000 DSC.
Typically less than 5 mg of material contained in a standard
aluminium pan fitted with a lid was heated over the temperature
range 25.degree. C. to 300.degree. C. at a constant heating rate of
10.degree. C. per minute. A purge gas using nitrogen was used--flow
rate 50 ml per minute.
Intermediate 1
5-Hydroxy-4-methoxy-2-nitrobenzoic Acid
##STR00005##
[0215] 4,5-Dimethoxy-2-nitrobenzoic acid (145 g, 0.639 mol) was
dissolved in a solution of sodium hydroxide (6N, 600 mL) and heated
at 100.degree. C. for 3 h. The mixture was cooled to RT, and poured
into a mixture of concentrated hydrochloric acid and crushed ice
(pH<2). The mixture was filtered, and the filter cake was dried
to give Intermediate 1 (149 g, crude) as a yellow solid, which was
used without further purification. .sup.1H NMR (DMSO-d.sub.6 400
MHz): .delta.7.34 (s, 1H), 6.89 (s, 1H), 3.80 (s, 3H).
Intermediate 2
2-Amino-5-hydroxy-4-methoxybenzoic Acid
##STR00006##
[0217] A mixture of Intermediate 1 (50 g, 93.85 mmol) and 10% Pd/C
(5 g) in MeOH (1.2 L) was stirred under H.sub.2 atmosphere (50 psi)
at RT for 4 h. The mixture was filtered and washed with MeOH
(10.times.1 L). The combined MeOH extracts were concentrated to
afford Intermediate 2 (27.7 g, 64% yield) as black solid which was
used without further purification.
Intermediate 3
7-Methoxyquinazoline-4,6-diol
##STR00007##
[0219] To a suspension of Intermediate 2 (88 g, 0.48 mol) in
2-methoxyethanol (2 L) was added formamidine (101 g, 0.96 mol) and
the reaction mixture was refluxed overnight. The reaction mixture
was concentrated, diluted with water (1.5 L) and neutralized (to
pH=7) with ammonia. The mixture was filtered and the precipitate
was washed with water. The precipitate was dried under reduced
pressure to afford Intermediate 3 as a brown solid (62 g, 67%
yield). .sup.1H NMR (DMSO-d.sub.6 400 MHz): .delta. 7.89 (s, 1H),
7.36 (s, 1H), 7.08 (s, 1H), 3.88 (s, 3H).
Intermediate 4
4-Hydroxy-7-methoxyquinazolin-6-yl acetate
##STR00008##
[0221] To a suspension of Intermediate 3 (52 g, 0.27 mol) and
pyridine (53.6 g, 0.68 mol) in anhydrous DCM (1 L) was added acetic
chloride (52.9 g, 0.68 mol) drop-wise and the mixture was stirred
overnight at RT. The mixture was poured into water (1 L) and
extracted with DCM several times. The combined organic layers were
washed with brine, dried over Na.sub.2SO.sub.4, concentrated to
afford Intermediate 4 as a black solid (63.2 g, 100% yield). 1H NMR
(DMSO-d.sub.6 400 MHz): .delta. 8.62 (s, 1H), 7.88 (s, 1H), 7.37
(s, 1H), 3.95 (s, 3H), 2.74 (s, 3H).
Intermediate 5
4-Chloro-7-methoxyquinazolin-6-yl acetate
##STR00009##
[0223] A suspension of Intermediate 4 (75.6 g, 0.323 mol) in
POCl.sub.3 (287 mL) was heated to refluxed for 0.5 h. The reaction
mixture was concentrated and diluted with DCM (500 mL), poured into
water (500 mL), filtered and washed with DCM. The combined organic
layers were washed with brine, dried over Na.sub.2SO.sub.4 and
concentrated. Purification by chromatography (PE/EtOAc=1/1) gave
Intermediate 5 (55 g, 67% yield) as white solid. .sup.1H NMR
(CDCl.sub.3 400 MHz): .delta. 8.95 (s, 1H), 7.90 (s, 1H), 7.43 (s,
1H), 4.02 (s, 1H), 2.39 (s, 1H).
Intermediate 6
4-[(3-Chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
acetate
##STR00010##
[0225] To a suspension of Intermediate 5 (100 g, 0.396 mol) in
acetonitrile (4 L) was added 2-fluoro-3-chloroaniline (60.5 g,
0.416 mol) and the reaction mixture was heated to 80.degree. C.
overnight. The precipitate was collected by filtration and dried in
vacuo to afford Intermediate 6 (181 g, 80% purity) as white solid
which was used for next step directly without purification. .sup.1H
NMR (DMSO-d.sub.6 400 MHz): .delta. 8.93 (s, 1H), 8.82 (s, 1H),
7.67-7.63 (m, 1H), 7.59 (s, 1H), 7.56-7.52 (m, 1H), 7.39-7.35 (m,
1H), 4.02 (s, 3H), 2.39 (s, 3H).
Intermediate 7
4-[(3-Chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-ol
##STR00011##
[0227] To a solution of Intermediate 6 (181 g, 0.396 mol) in MeOH
(2 L) was added potassium carbonate (138 g, 1 mol) and the reaction
mixture was stirred at RT overnight. The reaction mixture was
filtered and the solid washed with MeOH. The filtrate was
concentrated in vacuo to afford Intermediate 7 (280 g, 60% purity,
contained potassium carbonate). .sup.1H NMR (DMSO-d.sub.6 400 MHz):
.delta. 8.01 (s, 1H), 7.61-7.58 (m, 1H), 7.27-7.24 (m, 1H),
7.17-7.13 (m, 1H), 6.95 (s, 1H), 6.83 (s, 1H), 3.79 (s, 3H).
Intermediate 8
tert-Butyl
(3R)-4-(chlorocarbonyl)-3-methylpiperazine-1-carboxylate
##STR00012##
[0229] To a mixture of triphosgene (23 g, 75 mmol) in anhydrous DCM
(250 mL) was added pyridine (18 g, 225 mmol) drop-wise followed by
addition of tert-butyl (3R)-3-methylpiperazine-1-carboxylate (15 g,
75 mmol) at 0.degree. C. The mixture was stirred overnight at RT.
TLC showed the starting material had been consumed. The mixture was
concentrated to afford Intermediate 8 as yellow solid, which was
used without further purification.
Intermediate 9
4-tert-Butyl
1-{4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl}(2R)-2-met-
hylpiperazine-1,4-dicarboxylate
##STR00013##
[0231] A mixture of Intermediate 7 (19.2 g, 60 mmol), Intermediate
8 prepared according to the above procedure and potassium carbonate
(16.6 g, 120 mmol) in anhydrous DMF (300 mL) was stirred overnight
at RT. The reaction mixture was poured into water (250 mL) and
filtered, and the filter cake was dried under vacuum to afford
Intermediate 9 (25 g, 75% yield) as yellow solid. HPLC:
t.sub.R=2.68 min in 10-80AB_6 min chromatography (Ultimate XB-C18,
3.0*50 mm, 3 um). LCMS: t.sub.R=0.792 min in 5-95AB_1.5 min
chromatography (Welch Xtimate C18, 2.1*30 mm, 3 um), MS (ESI) m/z
546.0 [M+H].sup.+.
Intermediate 10
4-[(3-Chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2-methylpiperazine-1-carboxylate
##STR00014##
[0233] A mixture of Intermediate 9 (8.3 g, 15 mmol) in DCM (100 mL)
and HCl/dioxane (10 mL, 4M) was stirred for 30 min at RT. After
filtration, the solid was collected and redissolved in water, and
then adjusted to pH=8 with saturated NaHCO.sub.3. The precipitate
was collected and washed with CH.sub.2Cl.sub.2. The solid was dried
under vacuum to give the Intermediate 10 (8 g, 85% purity) as
yellow solid. This crude product was used for the next step without
purification.
Intermediate 11
(S)-2,4-dimethylpiperazine-1-carbonyl chloride
##STR00015##
[0235] To a solution of triphosgene (1.04 g, 3.5 mmol) in DCM (20
mL) under nitrogen was added pyridine (2.3 g, 28.0 mmol) drop-wise
at 0.degree. C. followed by addition of (S)-1,3-dimethylpiperazine
(800 mg, 7.0 mmol) in DCM (30 mL), the reaction mixture was warmed
to RT and stirred overnight as monitored by TLC (R.sub.f=0.9, PE:
EtOAc=1:1). The mixture was concentrated to give Intermediate 11 (3
g, crude) which was used without purification.
Intermediate 12
(.+-.)-tert-Butyl
(4-(chlorocarbonyl)-3-methylpiperazine-1-carboxylate
##STR00016##
[0237] To a mixture of triphosgene (23 g, 75 mmol) in anhydrous DCM
(250 mL) was added pyridine (18 g, 225 mmol) drop-wise followed by
addition of (.+-.)-tert-butyl 3-methylpiperazine-1-carboxylate (15
g, 75 mmol) at 0.degree. C. The mixture was stirred overnight at
RT. TLC showed the starting material was consumed. The mixture was
concentrated to afford Intermediate 12 as yellow solid, which was
used without further purification.
Intermediate 13
(.+-.)-4-tert-Butyl
1-{4-[(2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl}2-methylpiperazine--
1,4-dicarboxylate
##STR00017##
[0239] A mixture of Intermediate 7 (19.2 g, 60 mmol), Intermediate
12 prepared according to above procedure and potassium carbonate
(16.6 g, 120 mmol) in anhydrous DMF (300 mL) was stirred overnight
at RT. The reaction mixture was poured into water (250 mL) and
filtered, and the filter cake was dried under vacuum to afford
Intermediate 13 (25 g, 75% yield) as yellow solid. HPLC:
t.sub.R=2.68 min in 10-80AB_6 min chromatography (Ultimate XB-C18,
3.0*50 mm, 3 um). LCMS: t.sub.R=0.792 min in 5-95AB_1.5 min
chromatography (Welch Xtimate C18, 2.1*30 mm, 3 um), MS (ESI) m/z
546.0 [M+H].sup.+.
Intermediate 14
(.+-.)-4-[(3-Chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl-2-methy-
lpiperazine-1-carboxylate
##STR00018##
[0241] A mixture of Intermediate 13 (25 g, 46 mmol) in a solution
of HCl/dioxane (250 mL, 4M) was stirred for 30 min at RT. The
resulting solid was collected and redissolved in water, and then
adjusted to pH=8 with saturated NaHCO.sub.3. The precipitate was
collected and washed with CH.sub.2Cl.sub.2. The solid was dried
under vacuum to give the product (19 g, 93% yield) as yellow solid.
HPLC: t.sub.R=1.58 min in 10-80AB_6 min chromatography (Ultimate
XB-C18, 3.0*50 mm, 3 um). LCMS: t.sub.R=0.638 min in 5-95AB_1.5 min
chromatography (Welch Xtimate C18, 2.1*30 mm, 3 um), MS (ESI) m/z
445.1 [M+H].sup.+. .sup.1H NMR (CD.sub.3OD 400 MHz): .delta. 8.44
(s, 1H), 8.08 (s, 1H), 7.60 (t, 1H), 7.39 (t, 1H), 7.27-7.20 (m,
2H), 4.41 (s, 1H), 4.00 (s, 3H), 3.08-2.79 (m, 4H), 1.43 (brs,
3H).
Example 1
4-[(3-Chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate
##STR00019##
[0243] To a mixture of Intermediate 10 (8 g, 15 mmol, 85% purity)
and paraformaldehyde (1 g, 32 mmol) in MeOH (100 mL) was added
sodium cyanoborohydride (2 g, 32 mmol) and the reaction mixture was
stirred at RT overnight. The reaction mixture was concentrated in
vacuo, the residue was diluted with water and extracted with EtOAc
(3.times.100 mL). The combined organic layers were washed with
brine, dried over anhydrous sodium sulfate and concentrated under
reduced pressure. The crude product was purified by reverse phase
preparative HPLC (column: synergi 77*250, 10 um, gradient: 5-35% B
(A=water/0.05% formic acid, B=acetonitrile), flow rate: 140
mL/min). The fraction contained desired product was neutralized
with saturated potassium carbonate and extracted with EtOAc. The
combined organic layer was concentrated in vacuo and freeze-dried
to afford Example 1 (4 g, 58% yield for 2 steps) as white
solid.
[0244] LC-MS: t.sub.R=1.406 min in 4 min chromatography, MS (ESI)
m/z 460.0 [M+H].sup.+
[0245] SFC: t.sub.R=1.637 min in 3 min chromatography (Chiralpak
AD-3 50*4.6 mm I.D, 3 um), MS (ESI) m/z 460.1 [M+H].sup.+
[0246] .sup.1H NMR (CDCl.sub.3 400 MHz): .delta. 8.76 (s, 1H),
8.53-8.48 (m, 1H), 7.65 (s, 1H), 7.44 (brs, 1H), 7.34 (s, 1H),
7.19-7.15 (m, 2H), 4.51-4.50 (m, 1H), 4.20-4.05 (m, 1H), 3.99 (s,
3H), 3.50-3.30 (m, 1H), 2.87 (d, 1H), 2.73 (d, 1H), 2.35 (s, 3H),
2.35-2.25 (m, 1H), 2.13-2.11 (m, 1H), 1.47 (s, 3H).
Example 1, Form A
[0247] Form A material was produced by heating Example 1 to
140.degree. C. Approximately 10 mg of Example 1 was placed in an
aluminium pan. The pan was heated to 140.degree. C. with the
heating rate of 10.degree. C./min using differential scanning
calorimetry (DSC) and subsequently cooled to RT under nitrogen
gas.
[0248] Form A material was also produced by slow evaporation of
Example 1 in IPA. Approximately 10 mg of Example 1 was weighed to a
3-mL vial, 0.25 mL of IPA was added to dissolve the solid. After
evaporating at RT for 24 hours, Example 1 (Form A) was
obtained.
[0249] Form A material was also produced by slurrying Example 1 in
MTBE for 24 hours at 50.degree. C. Approximately 10 mg of Example 1
was weighed to a 3-mL vial, 1 mL of MTBE was added and then the
suspension was stirred for 24 hours at 50.degree. C. to obtain
Example 1 (Form A) was obtained.
[0250] Form A material was also produced by anti-solvent addition
of EtOAc/heptane. Approximately 10 mg of Example 1 was weighed to a
5-mL vial, 1 mL of EtOAc was added to dissolve the solid and the 4
mL of anti-solvent heptane was added to the vial slowly. The
mixture was stirred for 24 hours at RT to obtain Example 1 (Form
A).
[0251] The X-ray powder diffraction spectra for Example 1 (Form A)
showed the material to be crystalline. The material had a melting
point of 192.4.degree. C. (onset).
Example 1, Form E
[0252] Approximately 10 mg of Example 1 was weighed to a 5 mL vial,
0.25 mL of THF was added to dissolve the solid, then 4 mL of
anti-solvent heptane was added to the vial and the mixture stirred
for 24 hours at RT before the solid was isolated. The sample (Form
E) was determined to be crystalline by XRPD and had a melting point
of 194.2.degree. C. (onset).
Example 1, Form I
[0253] Approximately 10 mg of Example 1 was weighed to a 3 mL vial,
1 mL of H.sub.2O to was added the vial and the suspension stirred
for 24 hours at 50.degree. C. before the solid was isolated. The
sample (Form I) was determined to be crystalline by XRPD and had a
melting point of 193.3.degree. C. (onset).
Example 1, Form J
[0254] Approximately 10 mg of Example 1 was weighed to a 3 mL vial,
1 mL of H.sub.2O was added to the vial and the suspension stirred
for 24 hours at RT before the solid was isolated. The sample (Form
J) was determined to be crystalline by XRPD and had a melting point
of 193.3.degree. C. (onset).
Example 2
4-[(3-Chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate hydrochloride
##STR00020##
[0256] Example 1 (1.8 g) was dissolved in acetonitrile (5 mL), then
1 N HCl (5 mL) was added slowly, the solution was dried by
hyophilization to give Example 2 (1.93 g) as a yellow solid. LC-MS:
t.sub.R=1.355 min in 4 min chromatography, MS (ESI) m/z 460.1
[M+H].sup.+. SFC: t.sub.R=1.773 min in 3 min chromatography
(Chiralpak AD-3 50*4.6 mm I.D, 3 um), MS (ESI) m/z 460.1
[M+H].sup.+. .sup.1H NMR (CD.sub.3OD 400 MHz): .delta. 8.55 (s,
1H), 8.33-8.16 (m, 1H), 7.56 (t, 1H), 7.45 (t, 1H), 7.33 (s, 1H),
7.28-7.20 (m, 1H), 4.81-4.59 (m, 1H), 4.52-4.15 (m, 1H), 4.10-3.95
(m, 3H), 3.74-3.48 (m, 3H), 3.35 (br. s., 1H), 3.24-3.09 (m, 1H),
2.97 (s, 3H), 1.54 (br. s., 3H). [.alpha.].sub.D.sup.25=-14.96
(c10, DMSO).
Formation of Example 2 Mono-HCl Salt Form A.sub.1
[0257] To approximately 10 mg of Example 1 was added 0.35 mL of
IPA, followed by 0.217 mL of hydrochloric acid. The solution was
sealed tightly with a cap and left to stir on a magnetic stirrer
plate. During the stirring, some white precipitate was observed.
After approximately 24 hours, the sample was separated and dried at
RT by vacuum. This form (mono-HCl salt Form A.sub.1) was determined
to be crystalline by XRPD and had a melting point of 259.6.degree.
C. (onset).
[0258] Mono-HCl salt Form A.sub.1 was also produced by reaction
crystallization of Example 1 and hydrochloric acid in EtOH at RT.
To approximately 10 mg of Example 1, was added 0.35 mL of EtOH to
dissolve the solid, then 0.217 mL of hydrochloric acid was added to
the solution. The solution was sealed tightly with a cap and left
to stir on a magnetic stirrer plate. During the stirring, some
white precipitate was observed. After approximately 24 hours, the
sample was separated and dried at RT by vacuum. This form (mono-HCl
salt Form A.sub.1) was determined to be crystalline by XRPD and had
a melting point of 259.6.degree. C. (onset).
[0259] Mono-HCl salt Form A.sub.1 was also produced by reaction
crystallization of Example 1 and hydrochloric acid in acetone at
RT. To approximately 10 mg of Example 1, was added 0.35 mL of
acetone to dissolve the solid, followed by 0.217 mL of hydrochloric
acid. The solution was sealed tightly with a cap and left to stir
on a magnetic stirrer plate. During the stirring, some white
precipitate was observed. After approximately 24 hours, the sample
was separated and dried at RT by vacuum. This form (mono-HCl salt
Form A.sub.1) was determined to be crystalline by XRPD and had a
melting point of 259.6.degree. C. (onset).
[0260] Mono-HCl salt Form A.sub.1 was also produced by reaction
crystallization of Example 1 and hydrochloric acid in THF at RT. To
approximately 10 mg of Example 1, was added 0.35 mL of THF to
dissolve the solid, then 0.217 mL of hydrochloric acid was added.
The solution was sealed tightly with a cap and left to stir on a
magnetic stirrer plate. During the stirring, some white precipitate
was observed. After approximately 24 hours, the sample was
separated and dried at RT by vacuum. This form (mono-HCl salt Form
A.sub.1) was determined to be crystalline by XRPD and seen to be
different to previously seen forms. This material had a melting
point of 259.6.degree. C. (onset).
Example 3
4-[(3-Chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2S)-2,4-dimethylpiperazine-1-carboxylate
##STR00021##
[0262] A solution of Intermediate 7 (150 mg, 0.47 mmol),
Intermediate 11 (1 g, crude) and K.sub.2CO.sub.3 (130 mg, 0.94
mmol) in N,N-dimethyl-formamide (10 mL) was stirred at 30.degree.
C. overnight as monitored by LCMS. The solution was filtered and
purified by reverse phase preparative HPLC (column: ASB 150*25 mm*5
um, gradient: 3-28% B (A=water/0.05% HCl, B=acetonitrile), flow
rate: 30 mL/min) to give Example 3 (21.0 mg). LC-MS t.sub.R=1.156
min in 4 min chromatography, MS (ESI) m/z 460.0 [M+H].sup.+ SFC:
t.sub.R=2.084 min in 3 min chromatography (Chiralpak AD-3 50*4.6 mm
I.D, 3 um), MS (ESI) m/z 460.1 [M+H].sup.+; .sup.1H NMR
(CD.sub.3OD, 400 MHz): .delta.8.77 (s, 1H), 8.43 (s, 1H), 7.57-7.50
(m, 2H), 7.38 (s, 1H), 7.32-7.28 (m, 1H), 4.51-4.21 (m, 1H), 4.10
(s, 3H), 3.77-3.35 (m, 5H), 3.27-3.17 (m, 1H), 2.99 (s, 3H),
1.58-1.49 (m, 3H).
Example 4
4-[(3-Chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl (.+-.)
2,4-dimethylpiperazine-1-carboxylate
##STR00022##
[0264] A mixture of Intermediate 14 (1.0 g, 2.0 mmol, 96% purity),
paraformaldehyde (200 mg, 6.6 mmol), acetic acid (400 mg, 6.6 mmol)
in MeOH (15 mL) was stirred for 2 hours at RT. Sodium
cyanoborohydride (400 mg, 6.6 mmol) was added. The resulting
reaction mixture was stirred for another 2 hours. The mixture was
worked up and purified by reverse phase preparative HPLC (column:
ASB, gradient: 5-30% B (A=water/0.05% HCl, B=acetonitrile), flow
rate: 30 mL/min) to afford Example 4 (300 mg, 27%) as white solid.
LC-MS t.sub.R=1.099 min in 4 min chromatography, MS (ESI) m/z 460.1
[M+H]+; .sup.1H NMR (CD.sub.3OD 400 MHz): .delta. 8.79 (s, 1H),
8.51 (s, 1H), 7.58-7.52 (dd, 2H), 7.45 (s, 1H), 7.34-7.30 (t, 1H),
4.71-4.30 (m, 2H), 4.13 (s, 3H), 3.75-3.58 (m, 3H), 3.55-3.42 (m,
1H), 3.27 (s, 1H), 3.02 (s, 3H), 1.62-1.53 (m, 3H).
Example 5
Alternative crystalline forms of
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate
TABLE-US-00009 [0265] XRPD Angle 2- XRPD Melting Freebase Theta
Intensity point Form Preparation Methods (2.theta.) (%) (.degree.
C.) Form A Amorphous freebase heated 23.3, 100.00, 192.4 to
135.degree. C. (10.degree. C./min) 14.3, 83.70, Slow evaporation:
IPA, 9.4 78.08 iPrOAc Slurry in heptane, MTBE, DCM/heptane (1/4,
v/v), THF/heptane (1/4, v/v), iPrOAc/heptane (1/4, v/v) at RT
Slurry in heptane, MTBE, acetone/H.sub.2O (1/4, v/v), MeOH/H.sub.2O
(1/4, v/v), EtOH/H.sub.2O (1/4, v/v), THF/H.sub.2O (1/4, v/v),
DCM/heptane (1/4, v/v), THF/heptane (1/4, v/v), iPrOAc/heptane
(1/4, v/v), EtOH/heptane (1/4, v/v) at 50.degree. C. Anti-solvent
addition (solvent/anti-solvent): EtOAc/heptane, DCM/heptane Wet
grinding: acetone, EtOAc Form B Slurry in acetone/H.sub.2O 6.3,
100.00, N/A (1/4, v/v), 3.1, 52.07, MeOH/H.sub.2O (1/4, v/v), 12.6
35.29 EtOH/H.sub.2O (1/4, v/v), THF/H.sub.2O (1/4, v/v) at RT
Anti-solvent addition (solvent/anti-solvent): MeOH/H.sub.2O,
THF/H.sub.2O, Dioxane/H.sub.2O Wet grinding: EtOH/H.sub.2O (1/1,
v/v) Form C Slow evaporation: THF 15.6, 100.00, N/A Anti-solvent
addition 8.6, 60.20, (solvent/anti-solvent): 13.9 34.59
dioxane/hepane Form D Slow evaporation: EtOH 7.3, 100.00, N/A
Slurry in EtOH/heptane 11.4, 39.48, (1/4, v/v) at RT 21.0 23.59
Anti-solvent addition (solvent/anti-solvent): EtOH/MTBE Form E
Anti-solvent addition 7.3, 100.00, 93.0 (solvent/anti-solvent):
13.7, 81.83, THF/heptane 13.4 74.07 Form F Slow evaporation: 9.3,
100.00, N/A acetone 16.0, 69.50, 21.6 57.55 Form G Slow
evaporation: 5.1, 100.00, N/A acetone 7.2, 12.14, Wet grinding: DCM
17.0 8.13 Form H Slow evaporation: MeOH 7.7, 100.00, N/A Wet
grinding: MeOH 21.2, 41.70, 19.5 39.40 Form I Slurry in H.sub.2O at
50.degree. C. 3.5, 100.00, 193.3 7.0, 41.22, 9.5 32.57 Form J
Slurry in H.sub.2O at RT 7.8, 100.00, N/A 7.0, 49.36, 4.9 45.57
Example 6
Alternative Salt Forms of
4-[(3-chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate
TABLE-US-00010 [0266] XRPD Angle 2- XRPD Melting Salt Theta
Intensity point Form Preparation Methods (2.theta.) (%) (.degree.
C.) HCl salt Reaction crystallization 12.3, 100.00, 259.6 Form
A.sub.1 of the freebase and 13.9, 40.45, hydrochloric acid in 9.3
29.34 IPA, EtOH, acetone or THF at RT HCl salt Reaction
crystallization 6.6, 100.00, N/A Form B.sub.1 of the freebase and
13.2, 52.30, hydrochloric acid in 12.6 38.68 EtOH/H.sub.2O (v/v,
19/1) at RT, then evaporation Sulfate Reaction crystallization
19.8, 100.00, N/A Form A.sub.2 of the freebase and 20.4, 36.69,
sulfuric acid in IPA at 22.3 26.58 RT Sulfate Reaction
crystallization 7.2, 100.00, 223.7 Form B.sub.2 of the freebase and
16.7, 68.32, sulfuric acid in EtOH, 14.5 45.68 acetone, THF or
EtOH/H.sub.2O (v/v, 19/1)at RT Phosphate Reaction crystallization
7.0, 100.0, 206.0 Form A.sub.3 of the freebase and 16.5, 61.81,
phosphoric acid in EtOH 22.4 29.12 at RT Phosphate Reaction
crystallization 5.1, 100.00, 177.8 Form B.sub.3 of the freebase and
23.4, 18.27, phosphoric acid in 11.9 16.19 EtOH/H.sub.2O (v/v,
19/1) at RT Maleate Reaction crystallization 4.9, 100.00, 108.1
Form A.sub.4 of the freebase and 6.6, 93.00, maleic acid in IPA at
RT 12.6 30.51 Reaction crystallization of the freebase and maleic
acid in acetone at RT, then evaporation Maleate Reaction
crystallization 6.7, 100.00, 120.0 Form B.sub.4 of the freebase and
4.5, 26.67, maleic acid in DCM or 20.2 11.01 THF at RT Maleate
Reaction crystallization 6.3, 100.00, N/A Form C.sub.4 of the
freebase and 8.5, 87.86, maleic acid in EtOH/H.sub.2O 10.6 63.25
(v/v, 19/1) at RT, then evaporation Tartrate Reaction
crystallization 13.3, 100.00, 158.5 Form A.sub.5 of the freebase
and 6.6, 63.41, tartaric acid in EtOH or 17.6 49.61 EtOH/H.sub.2O
(v/v, 19/1) at RT Fumarate Reaction crystallization 6.6, 100.00,
212.8 Form A.sub.6 of the freebase and 5.2, 51.69, fumaric acid in
acetone 20.4 29.49 at RT Reaction crystallization of the freebase
and fumaric acid in IPA at RT, then evaporation Fumarate Reaction
crystallization 9.3, 100.00, 205.8 Form B.sub.6 of the freebase and
9.8, 58.74, fumaric acid in DCM at 26.7 54.18 RT Fumarate Reaction
crystallization 7.2, 100.00, 199.2 Form C.sub.6 of the freebase and
17.0, 86.58, fumaric acid in EtOH/H.sub.2O 6.2 54.86 (v/v, 19/1) at
RT, then evaporation Citrate Reaction crystallization 28.3, 100.00,
157.9 Form A.sub.7 of the freebase and 15.2, 36.42, citric acid in
DCM at RT 22.2 26.50 Succinate Reaction crystallization 6.5,
100.00, 191.8 salt of the freebase and 17.7, 31.30, Form A.sub.8
succinic acid in 14.7 24.91 acetone, DCM or EtOAc at RT Reaction
crystallization of the freebase and succinic acid in EtOH at RT,
then evaporation Succinate Reaction crystallization 6.0, 100.00,
N/A salt of the freebase and 24.3, 70.58, Form B.sub.8 succinic
acid in 8.3 62.41 acetone/H.sub.2O (v/v, 19/1) at RT, then
evaporation Adipate Reaction crystallization 5.0, 100.00, 133.4
Form A.sub.9 of the freebase and 8.5, 9.38, adipic acid in DCM at
RT 16.6 6.46 Reaction crystallization of the freebase and adipic
acid in acetone or EtOAc at RT, then evaporation Adipate Reaction
crystallization 4.4, 100.00, N/A Form B.sub.9 of the freebase and
6.2, 43.19, adipic acid in EtOH at 15.7 36.16 RT, then evaporation
Mesylate Reaction crystallization 13.1, 100.0, N/A Form A.sub.10 of
the freebase and 16.9, 72.46, methanesulfonic acid in 7.2 63.66
acetone at RT Mesylate Reaction crystallization 18.6, 100.00, 224.0
Form B.sub.10 of the freebase and 23.0, 94.62, methanesulfonic acid
in 19.3 86.41 DCM or EtOAc at RT Malonate Reaction crystallization
15.0, 100.00, 157.7 Form A.sub.11 of the freebase and 13.1, 65.81,
malonic acid in DCM or 10.2 50.20, EtOAc at RT Reaction
crystallization of the freebase and malonic acid in acetone or EtOH
at RT, then evaporation Benzoate Reaction crystallization 5.9,
100.00, N/A Form A.sub.12 of the freebase and 17.0, 38.62, benzoic
acid in acetone, 3.7 22.28 THF, EtOH, DCM or acetone/H.sub.2O (v/v,
19/1) at RT, then evaporation Benzoate Reaction crystallization
6.0, 100.00, N/A Form B.sub.12 of the freebase and 26.3, 56.07,
benzoic acid in EtOAc at 25.9 47.79 RT, then evaporation
Example 7
4-[(3-Chloro-2-fluorophenyl)amino]-7-methoxyquinazolin-6-yl
(2R)-2,4-dimethylpiperazine-1-carboxylate succinate
##STR00023##
[0268] Example 1 (10 mg, 0.022 mmol) was dissolved in 0.44 mL of
acetone in a vial. Succinic acid (2.57 mg, 0.022 mmol) was added to
the solution. The resulting mixture was sealed tightly with a cap
and allowed to stir on a magnetic stirrer plate. During the
stirring, some white precipitate was observed. After approximately
24 hours, the white solid was separated and dried at room
temperature under vacuum. .sup.1H NMR (400 MHz, DMSO-d.sub.6) 9.74
(s, 1H), 8.47 (s, 1H), 8.22 (s, 1H), 7.51-7.47 (m, 2H), 7.34 (s,
1H), 7.30-7.26 (m, 1H), 4.4-4.2 (br, 1H), 3.95 (s, 3H), 3.9-3.7
(br, 1H), 2.82-2.80 (d, 1H), 2.70-2.67 (s, 1H), 2.42 (s, 2H), 2.21
(s, 3H), 2.12-2.10 (m, 1H), 1.94-1.89 (m, 1H), 1.34 (s, 3H).
Formation of Example 7 Succinate Salt Form A.sub.8
[0269] Succinate salt Form A.sub.8 was produced by the procedure
described above. This form (Succinate salt Form A.sub.8) was
determined to be crystalline by XRPD and had a melting point of
191.8.degree. C. (onset).
[0270] Succinate salt Form A.sub.8 was also produced by reaction
crystallization of Example 1 and succinic acid in EtOH at RT. To
approximately 10 mg of Example 1, was added 0.59 mL of EtOH to
dissolve the solid, then 2.57 mg of succinic acid was added to the
solution. The solution was sealed tightly with a cap and left to
stir on a magnetic stirrer plate. After approximately 24 hours
stirring, the solution was evaporated to dryness at RT. This form
(Succinate salt Form A.sub.8) was determined to be crystalline by
XRPD and had a melting point of 191.8.degree. C. (onset).
[0271] Succinate salt Form A.sub.8 was also produced by reaction
crystallization of Example 1 and succinic acid in DCM at RT. To
approximately 10 mg of Example 1, was added 0.25 mL of DCM to
dissolve the solid, then 2.57 mg of succinic acid was added. The
solution was sealed tightly with a cap and left to stir on a
magnetic stirrer plate. During the stirring, some white precipitate
was observed. After approximately 24 hours, the sample was
separated and dried at RT by vacuum. This form (Succinate salt Form
A.sub.8) was determined to be crystalline by XRPD and had a melting
point of 191.8.degree. C. (onset).
Succinate salt Form A.sub.8 was also produced by reaction
crystallization of Example 1 and succinic acid in EtOAc at RT. To
approximately 10 mg of Example 1, was added 0.25 mL of EtOAc to
dissolve the solid, then 2.57 mg of succinic acid was added. The
solution was sealed tightly with a cap and left to stir on a
magnetic stirrer plate. During the stirring, some white precipitate
was observed. After approximately 24 hours, the sample was
separated and dried at RT by vacuum. This form (Succinate salt Form
A.sub.8) was determined to be crystalline by XRPD and had a melting
point of 191.8.degree. C. (onset).
* * * * *