U.S. patent number RE48,687 [Application Number 16/517,790] was granted by the patent office on 2021-08-17 for pyridinylaminopyrimidine derivatives, preparation process and use thereof.
This patent grant is currently assigned to Shanghai Allist Pharmaceuticals Co., Ltd.. The grantee listed for this patent is Shanghai Allist Pharmaceuticals Co., Ltd.. Invention is credited to Huibing Luo, Shuhui Wang, Yong Wu, Huayong Zhou.
United States Patent |
RE48,687 |
Luo , et al. |
August 17, 2021 |
Pyridinylaminopyrimidine derivatives, preparation process and use
thereof
Abstract
The present invention relates to pyridinylaminopyrimidine
derivatives represented by the following formula (I), and
pharmaceutically acceptable salts, preparation process and use
thereof, wherein R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, m and
A are defined as in the description. Pyridinylaminopyrimidine
derivatives of the present invention can selectively inhibit the
activity of mutant-type epidermal growth factor receptor (EGFR),
have a good inhibition for the cancer cell proliferation, and
therefore can be used as a therapeutic agent for treating tumors
and relevant diseases. ##STR00001##
Inventors: |
Luo; Huibing (Shanghai,
CN), Zhou; Huayong (Shanghai, CN), Wang;
Shuhui (Shanghai, CN), Wu; Yong (Shanghai,
CN) |
Applicant: |
Name |
City |
State |
Country |
Type |
Shanghai Allist Pharmaceuticals Co., Ltd. |
Shanghai |
N/A |
CN |
|
|
Assignee: |
Shanghai Allist Pharmaceuticals
Co., Ltd. (Shanghai, CN)
|
Family
ID: |
1000005472929 |
Appl.
No.: |
16/517,790 |
Filed: |
July 22, 2019 |
PCT
Filed: |
July 29, 2015 |
PCT No.: |
PCT/CN2015/000540 |
371(c)(1),(2),(4) Date: |
January 25, 2017 |
PCT
Pub. No.: |
WO2016/015453 |
PCT
Pub. Date: |
April 02, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
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Reissue of: |
15329044 |
Jul 29, 2015 |
10072002 |
Sep 11, 2018 |
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Foreign Application Priority Data
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Jul 29, 2014 [CN] |
|
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201410365911.4 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K
31/506 (20130101); C07D 401/14 (20130101); C07D
471/04 (20130101); C07D 403/14 (20130101) |
Current International
Class: |
C07D
471/04 (20060101); A61K 31/506 (20060101); C07D
403/14 (20060101); C07D 401/14 (20060101) |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
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102482277 |
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May 2012 |
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CN |
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103702990 |
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Apr 2014 |
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CN |
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104761544 |
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Jul 2015 |
|
CN |
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2012061299 |
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May 2012 |
|
WO |
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2013014448 |
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Jan 2013 |
|
WO |
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2015188777 |
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Dec 2015 |
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WO |
|
Other References
Office Action corresponding to Japanese Patent Application No.
2017-504644 dated Mar. 13, 2018. cited by applicant .
Ward et al. "Structure-and-Reactivity-Based Development of Covalent
Inhibitors of the Activating and Gatekeeper Mutant Forms of the
Epidermal Growth Factor Receptor (EGFR)", Journal of Medicinal
Chemistry, American Chemical Society 56(17):7025-7048 (2013). cited
by applicant .
Besse et al. "Neratinib (HKI-272), an irreversible pan-ErbB
receptor tyrosine kinase inhibitor: preliminary results of a phase
2 trial in patients with advanced non-small cell lung cancer",
Poster Session--Phase II 203:64 (2008). cited by applicant .
Janne et al. "Multicenter, Randomized, Phase II Trial of CI-1033,
an Irreversible Pan-ERBB Inhibitor, for Previously Treated Advanced
Non-Small-Cell Lung Cancer", J. Clin. Oncol. 25(25):3936-3944
(2007). cited by applicant .
Janne et al. "Phase I Dose-Escalation Study of the Pan-HER
Inhibitor, PF299804, in Patients with Advanced Malignant Solid
Tumors", Clin. Cancer Res. 17:1131-1139 (2011). cited by applicant
.
Katakami et al. "LUX-Lung 4: A Phase II Trial of Afatinib in
Patients with Advanced Non-Small-Cell Lung Cancer who Progressed
During Prior Treatment with Erlotinib, Gefitinib, or Both", J.
Clin. Oncol. 31:3335-3341 (2013). cited by applicant .
Landi et al. "Irreversible EGFR-TKIs: dreaming perfection", Transl
Lung Cancer Res. 2(1);40-49 (2013). cited by applicant .
Li et al. "BIBW2992, an irreversible EGFR/HER2 inhibitor highly
effective in preclinical lung cancer models", Oncogene 27:4702-4711
(2008). cited by applicant .
Pao et al. "Acquired Resistance of Lung Adenocarcinomas to
Gefitinib or Erlotinib is Associated with a Second Mutation in the
EGFR Kinase Domain", PLoS Medicine 2(3):0225-0235 (2005). cited by
applicant .
Walter et al. "Discovery of a mutant-selective covalent inhibitor
of EGFR that overcomes T790M-mediated resistance in NSCLC", Cancer
Discov. 3(12):1404-1415 (2013). cited by applicant .
Zhou et al. "Novel mutant-selective EGFR kinase inhibitors against
EGFR T790M", Nature 462(24):1070-1074 (2009). cited by applicant
.
International Search Report corresponding to International
Application No. PCT/CN2015/000540 dated Oct. 28, 2015. cited by
applicant .
Chinese Office Action corresponding to Chinese Application No.
201410365911.4 dated Jul. 25, 2017. cited by applicant .
Office Action corresponding to Canadian Application No. 2,956,628
dated Apr. 25, 2018. cited by applicant .
Extended European Search Report corresponding to European
Application No. 15828199.8 dated Nov. 29, 2017. cited by
applicant.
|
Primary Examiner: Johnson; Jerry D
Attorney, Agent or Firm: Myers Bigel, P.A.
Claims
What is claimed is:
1. A compound represented by the following general formula (I), or
a pharmaceutically acceptable salt thereof, ##STR00110## wherein,
Ring A is aryl or heteroaryl; R.sub.1 is selected from a group
consisting of hydrogen, halogen, C.sub.1-C.sub.4alkyl,
haloC.sub.1-C.sub.4alkyl, C.sub.2-C.sub.5alkenyl,
C.sub.2-C.sub.6alkynyl or --CN; R.sub.2 is .[.selected from a group
consisting of C.sub.1-C.sub.4alkyl, haloC.sub.1-C.sub.4alkyl,
C.sub.2-C.sub.6alkenyl, --(CH.sub.2).sub.cOR.sub.7,
--(CH.sub.2).sub.qNR.sub.7R.sub.7' or --(CH.sub.2) C(O)R.sub.7.].
.Iadd.trifluoroethyl.Iaddend.; R.sub.4 is ##STR00111## each R.sub.5
is .[.dependently.]. .Iadd.independently .Iaddend.halogen,
C.sub.1-C.sub.4alkyl, haloC.sub.1-C.sub.4alkyl,
C.sub.2-C.sub.6alkenyl, C.sub.2-C.sub.6alkynyl, --OR.sub.6,
--C(O)R.sub.7, --C(O)NR.sub.7R.sub.7', --OR.sub.7,
--NR.sub.7R.sub.7', --CN or --NO.sub.2; R.sub.3 is selected from a
group consisting of halogen, --CN, --NO.sub.2,
C.sub.1-C.sub.4alkyl, haloC.sub.1-C.sub.4alkyl, --C(O)R.sub.6,
--C(O)R.sub.7, --C(O)NR.sub.7R.sub.7', --OR.sub.7, --OR.sub.6,
--NHR.sub.7, --NR.sub.7--(C.sub.1-C.sub.4alkyl),
--NR.sub.7-(haloC.sub.1-C.sub.4alkyl),
--NR.sub.7(CH.sub.2).sub.nC(O)R.sub.6, --NR.sub.6R.sub.7,
--NR.sub.7-heterocycloalkyl, wherein said heterocycloalkyl is
unsubstituted or substituted with 1-2 substituents selected from
R.sub.7, or --NR.sub.7SO.sub.2R.sub.7, or heterocycloalkyl that is
unsubstituted or substituted with 1-3 substituents selected from
halogen, C.sub.1-C.sub.4alkyl, haloC.sub.1-C.sub.4alkyl,
--(CH.sub.2).sub.nOH, --NR.sub.7R.sub.7', --OR.sub.7 or
--C(O)R.sub.7; wherein, R.sub.6 is --(CH.sub.2).sub.qOR.sub.7,
--(CH.sub.2).sub.qNR.sub.7R.sub.7',
--(CH.sub.2).sub.qNR.sub.7C(O)R.sub.7,
--(CH.sub.2).sub.qC(O)R.sub.7 or
--(CH.sub.2).sub.qC(O)NR.sub.7R.sub.7'; R.sub.7 and R.sub.7' are
each independently hydrogen, C.sub.1-C.sub.4alkyl,
C.sub.2-C.sub.6alkenyl, C.sub.2-C.sub.6alkynyl or
haloC.sub.1-C.sub.4alkyl, or R.sub.7, R.sub.7' and the nitrogen
atom attached thereto are cyclized together to form a
heterocycloalkyl that is unsubstituted or substituted with 1-3
substituents selected from halogen, C.sub.1-C.sub.4alkyl,
haloC.sub.1-C.sub.4alkyl, --(CH.sub.2).sub.nOH, --NR.sub.7R.sub.7',
--OR.sub.7 or --C(O)R.sub.7; m is 1, 2 or 3; n is 0, 1, 2, 3 or 4;
q is 0, 1, 2, 3 or 4.
2. The compound according to claim 1 or a pharmaceutically
acceptable salt thereof, wherein Ring A is heteroaryl.
3. The compound according to claim 2 or a pharmaceutically
acceptable salt thereof, wherein Ring A is indolyl, indazolyl,
.[.pyrro[2,3-c]pyridinyl.]. .Iadd.pyrrolo[2,3-c]pyridinyl.Iaddend.,
.[.pyrro[3,2-c]pyridinyl.]. .Iadd.pyrrolo[3,2-c]pyridinyl.Iaddend.,
.[.pyrro[2,3-b]pyridinyl.]. .Iadd.pyrrolo[2,3-b]pyridinyl.Iaddend.,
.[.pyrro[3,2-b]pyridinyl.]. .Iadd.pyrrolo[3,2-b]pyridinyl.Iaddend.,
.[.pyrro[2,3-b]pyrazinyl.]. .Iadd.pyrrolo[2,3-b]pyrazinyl.Iaddend.,
indolin-2-onyl, pyridinyl, pyrazolyl or pyrimidinyl.
4. The compound according to claim 1 or a pharmaceutically
acceptable salt thereof, wherein R.sub.1 is hydrogen, halogen or
haloC.sub.1-C.sub.4alkyl.
.[.5. The compound according to claim 1 or a pharmaceutically
acceptable salt thereof, wherein R.sub.2 is C.sub.1-C.sub.4alkyl or
haloC.sub.1-C.sub.4alkyl..].
.[.6. The compound according to claim 5 or a pharmaceutically
acceptable salt thereof, wherein R.sub.2 is C.sub.2-C.sub.4alkyl or
haloC.sub.2-C.sub.4alkyl..].
.[.7. The compound according to claim 6 or a pharmaceutically
acceptable salt thereof, wherein R.sub.2 is isopropyl or
trifluoroethyl..].
8. The compound according to claim 1 or a pharmaceutically
acceptable salt thereof, wherein R.sub.4 is ##STR00112## R.sub.7
and R.sub.7' are each independently hydrogen or
C.sub.1-C.sub.4alkyl.
9. The compound according to claim 8 or a pharmaceutically
acceptable salt thereof, wherein R.sub.4 is ##STR00113## R.sub.7 is
hydrogen.
10. The compound according to claim 1 or a pharmaceutically
acceptable salt thereof, wherein R.sub.3 is selected from a group
consisting of halogen, --CN, --NO.sub.2, C.sub.1-C.sub.4alkyl,
haloC.sub.1-C.sub.4alkyl, --C(O)R.sub.7, --C(O)NR.sub.7R.sub.7',
--OR.sub.7, --NHR.sub.7, --NR.sub.7--(C.sub.1-C.sub.4alkyl),
--NR.sub.7(CH.sub.2).sub.nC(O)R.sub.6 or --NR.sub.6R.sub.7, or
heterocycloalkyl that is unsubstituted or substituted with 1-3
substituents selected from halogen, C.sub.1-C.sub.4alkyl,
haloC.sub.1-C.sub.4alkyl, --(CH.sub.2).sub.nOH, --NR.sub.7R.sub.7',
--OR.sub.7 or --C(O)R.sub.7; wherein, R.sub.6 is
--(CH.sub.2).sub.qOR.sub.7, --(CH.sub.2).sub.qNR.sub.7R.sub.7',
--(CH.sub.2).sub.qC(O)R.sub.7 or
--(CH.sub.2).sub.qC(O)NR.sub.7R.sub.7'; R.sub.7 and R.sub.7' are
each independently hydrogen, C.sub.1-C.sub.4alkyl or
haloC.sub.1-C.sub.4alkyl, or R.sub.7, R.sub.7' and the nitrogen
atom attached thereto are cyclized together to form a
heterocycloalkyl; n is 0, 1, 2, 3 or 4; q is 0, 1, 2, 3 or 4.
11. The compound according to claim 10 or a pharmaceutically
acceptable salt thereof, wherein R.sub.3 is --NR.sub.6R.sub.7, in
which R.sub.6 is --(CH.sub.2).sub.qNR.sub.7R.sub.7', R.sub.7 and
R.sub.7' are each independently hydrogen or C.sub.1-C.sub.4alkyl, q
is 2.
12. The compound according to claim 10 or a pharmaceutically
acceptable salt thereof, wherein R.sub.3 is a heterocycloalkyl
substituted by one substituent selected from halogen,
C.sub.1-C.sub.4alkyl, haloC.sub.1-C.sub.4alkyl or
--NR.sub.7R.sub.7', R.sub.7 and R.sub.7' are each independently
hydrogen or C.sub.1-C.sub.4alkyl.
13. The compound according to claim 12 or a pharmaceutically
acceptable salt thereof, wherein said heterocycloalkyl is
pyrrolidinyl.
14. The compound according to claim 1 or a pharmaceutically
acceptable salt thereof, wherein each R.sub.5 is .[.dependently.].
.Iadd.independently .Iaddend.halogen, C.sub.1-C.sub.4alkyl,
haloC.sub.1-C.sub.4alkyl, --OR.sub.7, --NR.sub.7R.sub.7', --CN or
--NO.sub.2, R.sub.7 and R.sub.7' are each independently hydrogen or
C.sub.1-C.sub.4alkyl, m is 1, 2 or 3.
15. The compound according to claim 14 or a pharmaceutically
acceptable salt thereof, wherein each R.sub.5 is .[.dependently.].
.Iadd.independently .Iaddend.halogen, C.sub.1-C.sub.4alkyl,
--OR.sub.7 or --NR.sub.7R.sub.7', R.sub.7 and R.sub.7' are each
independently hydrogen or C.sub.1-C.sub.4alkyl, m is 1, 2 or 3.
16. The compound according to claim 1 or a pharmaceutically
acceptable salt thereof, wherein said compound is selected from a
group consisting of:
.[.N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{5-c-
hloro-[4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylam-
ide;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[4-(1--
methyl-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylamide;.].
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-(2,2,2-trifluoroethoxyl)-5-
-{[4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]amino)pyridin-3-yl}acrylamide;
N-(2-{[2-(dimethylamino)ethyl](methyl)amino}-6-(2,2,2-trifluoroethoxyl)-5-
-{5-chloro-[4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}ac-
rylamide;
.[.N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-
-{[4-(1-methyl-5-fluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}ac-
rylamide;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[-
4-(1-methyl-5,6-difluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}a-
crylamide;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{-
5-chloro-[4-(1-methyl-6-fluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin--
3-yl}acrylamide;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{5-chloro-[-
4-(1-methyl-5,6-difluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}a-
crylamide;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{-
5-chloro-[4-(1-methyl-5-fluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin--
3-yl}acrylamide;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{5-fluoro-[-
4-(1-methyl-5-fluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acryl-
amide;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{5-fl-
uoro-[4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylami-
de;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{5-fluor-
o-[4-(1-methyl-5,6-difluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-y-
l}acrylamide;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[4-(1-meth-
yl-6-fluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylamide;.].
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-(2,2,2-trifluoroethoxyl)-5-
-{5-fluoro-[4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}ac-
rylamide; .Iadd.and.Iaddend.
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-(2,2,2-trifluoroethoxyl)-5-
-{[4-(1-methyl-5-fluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}ac-
rylamide.[.;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{5-chloro-[-
4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylamide
methanesulfonate;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{5-chloro-[-
4-(1-methyl-5,6-difluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}a-
crylamide methanesulfonate;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[4-(1-meth-
yl-5,6-difluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylamide
methanesulfonate;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[5-chloro--
4-(1-methyl-1H-pyrro[2,3-b]pyridin-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}-
acrylamide;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[5-chloro--
4-(1-methyl-1H-pyrro[2,3-b]pyridin-5-yl)pyrimidin-2-yl]amino}pyridin-3-yl}-
acrylamide;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[5-chloro--
4-(1-methyl-1H-pyrazol-4-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylamide;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[5-chloro--
2'-methoxy-(4,5'-bipyrimidine)-2-yl]amino}pyridin-3-yl}acrylamide;
and
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[5-chloro--
2'-amino-(4,5'-bipyrimidine)-2-yl]amino}pyridin-3-yl}acrylamide.]..
17. A process for preparing the compound represented by the general
formula (I) of claim 1, comprising the steps of: ##STR00114##
##STR00115## or ##STR00116## wherein ring A, R.sub.1, R.sub.2,
R.sub.3, R.sub.4, R.sub.5 and m are defined as in claim 1; L
represents .[.a leaving group, including.]. hydrogen, halogen or
##STR00117## compounds (a) and (b) are used as starting material,
and subjected to substitution under the catalysts to produce an
Intermediate 2; the Intermediate 2 and an Intermediate 1 are
subjected to substitution or coupling reaction to produce a
compound (c), the nitro group of the compound (c) is reduced to
produce a compound (d), the compound (d) is acylated to produce a
compound (I); or the Intermediate 2 and an Intermediate 1' are
subjected to substitution or coupling reaction to directly produce
a compound (I).
18. A pharmaceutical composition, comprising the compound
represented by formula (I) of claim 1 or a pharmaceutically
acceptable salt thereof and a pharmaceutically acceptable carrier,
excipient or diluent.
19. A method for treating an EGFR activating or resistant mutation
mediated lung cancer in a mammal, said method comprises
administering to a mammal the compound represented by formula (I)
of claim 1 or a pharmaceutically acceptable salt thereof, or a
pharmaceutical composition comprising a therapeutically effective
amount of a compound represented by formula (I) of claim 1 .Iadd.or
a pharmaceutically acceptable salt thereof .Iaddend.and a
pharmaceutically acceptable carrier, excipient or diluent.
20. A method for selectively inhibiting an EGFR activating or
resistant mutation over a .Iadd.wild-type .Iaddend.EGFR, said
method comprises contacting a biological sample with or
administering to a lung cancer patient the compound represented by
formula (I) of claim 1 or a pharmaceutically acceptable salt
thereof or a pharmaceutical composition containing the same.
21. The method of claim 19, wherein the mammal is a human.
.Iadd.22. A compound, wherein said compound is
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-(2,2,2-trifluoroethoxyl)-5-
-{[4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylamide
or a pharmaceutically acceptable salt thereof..Iaddend.
Description
RELATED APPLICATIONS
This application is a 35 U.S.C. .sctn. 371 national phase
application of and claims priority to PCT Application
PCT/CN2015/000540 filed Jul. 29, 2015, which claims priority to
Chinese Application No. 201410365911.4 filed Jul. 29, 2014, the
entire contents of which are incorporated herein by reference in
their entirety.
TECHNICAL FIELD
The present invention relates to pyridinylaminopyrimidine
derivatives, which selectively inhibit the activity of
mutation-type epidermal growth factor receptor (EGFR), a
pharmaceutically acceptable salt thereof, a process for preparing
the same, a pharmaceutical composition containing said derivative
and a pharmaceutically acceptable salt thereof, uses of said
derivative and a pharmaceutically acceptable salt thereof in
treating some mutation-type EGFR mediated diseases and in
manufacture of a medicament for treating some mutation-type EGFR
mediated diseases.
BACKGROUND
Cancer has been considered as a disease of the intracellular signal
transconducing system or signal transduction mechanism. The most
common cause of cancer is a series of defects, either in proteins,
when they are mutated, or in the regulation of the quantities of
the proteins in the cells such that they are over or under
produced. Mutations to the cell surface receptors, which usually
transduce the signals into the cells by means of tyrosine kinases,
can lead to activation of the kinase in the absence of ligand, and
passing of a signal which does not really exist. Alternatively,
many receptor tyrosine kinases can be overexpressed on the cell
surface leading to an inappropriately strong response to a weak
signal.
Epidermal cell growth factors receptors (EGFR) are identified as
one significant driving factor in the process for cellular growth
and proliferation. The epidermal cell growth factors receptors
family is composed of EGFR (Erb-B1), Erb-B2 (HER-2/neu), Erb-B3 and
Erb-B4. The epidermal cell growth factors receptors are concerned
in the process for most cancers, such as lung cancer, colon cancer
and breast cancer. The overexpression and mutation of EGFR have
been proved to be the leading risk factor for a breast cancer with
poor prognosis. Besides, it has been verified that each of the
above four members of the receptors family can aggregate with
another member into a heterodimer, and form a signal transduction
complex. Overexpression of one or more member(s) of this family in
a malignant tumor will result in a synergistic signal
transduction.
EGFR belongs to the protein tyrosine kinase (PTK) family. The
protein tyrosine kinase is an group of enzymes which catalyze the
transportation of phosphate groups from adenosine triphosphate
(ATP) to the tyrosine residue located in a protein substrate.
Protein tyrosine kinases function in normal cell growth. The
overexpression and mutation of EGFR may cause the activation of
receptors without ligands and the phosphorylation of some proteins,
and then the signal for cell division is produced. As a result,
EGFR may magnify the weak signal excessively by its own
tyrosine-kinase action, and render the overproliferation of
cells.
Specific PTK inhibitors as a potential anti-cancer therapeutic drug
are of wide concern. Typical representatives of currently market
available EGFR reversible inhibitors include Gefitinib, Erlotinib
and Lapatinib, and inhibit the EGFR wild-type and activating
mutations (e.g. Exon 19 deletion activating mutation, or L858R
activating mutation). Their structures are as follows, and are
respectively useful for treating non-small cell lung cancer (NSCLC)
and breast cancer. Clinical study proves gefitinib and erlotinib
have a favorable therapeutic effect on NSCLC patients with EGFR
exon 19 deletion or L858R mutation. However, their limitations are
that patients develop drug resistance after treatment, so that
inhibitors of this type are limited in their further clinical
applications. The study shows that 50% of resistance formed after
the treatment with gefitinib and erlotinib is associated with a
second mutation occurred in EGFR (T790M) (Pao W. et al., Plos Med.,
2:1-11, 2005). The therapeutic effect as reversible inhibitor is
lost.
##STR00002##
T790M is located at the entrance of the ATP binding pocket of EGFR,
and the size of its side chain directly affects the ability of EGFR
binding to ATP. The T790M mutation spatially inhibits the
interaction of the EGFR inhibitor and the ATP binding site,
increases the affinity of EGFR to ATP, and makes the cells
resistant to the EGFR inhibitors.
Compared to reversible EGFR inhibitors, irreversible EGFR
inhibitors have very prominent advantages. Irreversible EGFR
inhibitors can inhibit EGFR for a long time and are only limited by
the normal rate of receptor re-binding (also called reversion). It
is found that the irreversible EGFR inhibitor can covalently bind
to the cysteine residue (Cys797) of the EGFR by Michael addition
reaction and expand the binding sites of irreversible EGFR
inhibitors and the ATP, so that the resistance caused by the T790M
mutation can be overcame to some extent (Li D et al., Oncogene,
27:4702-4711, 2008). Currently market available irreversible EGFR
inhibitors include BIBW-2992 (Afatinib), those in development
include HKI-272 (Neratinib), EKB-569 (Pelitinib), PF00299804
(Dacomitinib) and the like, and their structures are as
follows.
##STR00003##
However, these irreversible EGFR inhibitors, which can inhibit EGFR
T790M, also have a large inhibition effect on the wild-type EGFR,
leading to severe side effects such as diarrhea, erythra, nausea,
anorexia, and weakness (Besse, B. et al. Eur. J. Cancer Suppl., 6,
64, abstr. 203, 2008; Janne, P. A. et al., J. Clin. Oncol., 25:
3936-3944, 2007). Accordingly although it is reported in the
literature that in the preclinical study, BIBW2992 (Afatinib) and
PF00299804 (Dacomitinib) show a significant antitumor activity and
can inhibit the activities of EGFR and EGFR T790M, however, due to
the occurrence of these adverse reactions, the clinical dose and
the effective blood drug concentration are limited in the clinical
course. Therefore, there is no remarkable progress for BIBW2992
(Afatinib) and PF00299804 (Dacomitinib) in overcoming the T790M
resistant mutation (Katakami N, Atagi S, Goto K, et al. [J].
Journal of Clinical Oncology, 2013, 31(27): 3335-3341.; Janne P A,
Boss D S, Camidge D R, et al. [J]. Clinical Cancer Research, 2011,
17(5): 1131-1139.; Landi L, Cappuzzo F. [J]. Translational Lung
Cancer Research, 2013, 2(1): 40-49.).
The above-mentioned reversible or irreversible EGFR inhibitors,
being currently marketed or under development, are mainly
quinazoline compounds. The currently reported quinazoline EGFR
inhibitors are the ATP competitive inhibitors of wild-type EGFR,
leading to the occurrence of some side-reaction. In 2009, a group
of pyrimidine-based irreversible EGFR inhibitors which are specific
to the EGFR T790M was reported by the researchers, and the
structures are shown below. Compared to the existing aniline
quinazoline EGFR inhibitors, these pyrimidine-based compounds have
a 30-100 fold higher inhibition activity for the EGFR T790M, and a
100 fold lower inhibition activity for the wild-type EGFR
(WenjunZhou et al., Nature, 462:1070-1074, 2009). However, these
pyrimidine-based compounds did not enter the clinical study
later.
##STR00004##
International Patent Application WO 2012/061299 A1 filed by Avila
Therapeutics discloses another series of pyrimidine-based
compounds, and the structures are shown below. The representative
compound is CO1686. It is reported in the literature that CO1686
can selectively act on the EGFR activating mutation and the T790M
resistant mutation, but have a weak inhibition effect on the
wild-type EGFR (Walter A O, Sjin R T T, Haringsma H J, et al. [J].
Cancer discovery, 2013, 3(12): 1404-1415.). Currently, this
compound is ready to enter Phase-II clinical stage.
##STR00005##
International Patent Application WO 2013/014448 A1 filed by
ASTRAZENECA AB also discloses a series of pyrimidine-based
compounds, and their structures are shown below. The representative
compound is AZD9291. This compound has a better inhibition effect
on the EGFR activating mutation and the T790M resistant mutation
than the wild-type EGFR, and is now in Phase I clinical stage.
##STR00006##
There is an urgent demand in the current anti-tumor field to
overcome the problems of the clinically common EGFR resistant
mutation (e.g. T790M mutation) and the toxic and side effects of
the existing EGFR inhibitors, i.e., develop more small molecule
inhibitors that show a higher inhibition effect on some activating
mutation and resistant mutation EGFRs and a lower inhibition effect
on the wild-type EGFR. During the study of the EGFR inhibitors, the
present inventors surprisingly discovered a group of
pyridinylaminopyrimidine derivatives, which have a remarkably
higher inhibition activity on the EGFR activating mutation (e.g.
Exon 19 deletion activating mutation, or L858R activating mutation)
and the T790M resistant mutation than the wild-type EGFR (WT EGFR),
and has good selectivity, low toxic and side effects, and good
safety. It is expected that this kind of inhibitors will have a
good therapeutic effect, can overcome the problems of drug
resistance and toxic/side effects, and accordingly may have good
development prospects.
SUMMARY OF THE INVENTION
The present invention provides a compound represented by the
following general formula (I), or a pharmaceutically acceptable
salt thereof:
##STR00007## wherein, Ring A is aryl or heteroaryl; R.sub.1 is
selected from a group consisting of hydrogen, halogen,
C.sub.1-C.sub.4alkyl, haloC.sub.1-C.sub.4alkyl,
C.sub.2-C.sub.6alkenyl, C.sub.2-C.sub.6alkynyl or --CN; R.sub.2 is
selected from a group consisting of C.sub.1-C.sub.4alkyl,
haloC.sub.1-C.sub.4alkyl, C.sub.2-C.sub.6alkenyl,
--(CH.sub.2).sub.qOR.sub.7, --(CH.sub.2).sub.qNR.sub.7R.sub.7' or
--(CH.sub.2).sub.qC(O)R.sub.7; R.sub.4 is
##STR00008## Each R.sub.5 is .[.dependently.]. .Iadd.independently
.Iaddend.halogen, C.sub.1-C.sub.4alkyl, haloC.sub.1-C.sub.4alkyl,
C.sub.2-C.sub.6alkenyl, C.sub.2-C.sub.6alkynyl, --OR.sub.6,
--C(O)R.sub.7, --C(O)NR.sub.7R.sub.7', --OR.sub.7,
--NR.sub.7R.sub.7', --CN or --NO.sub.2; R.sub.3 is selected from a
group consisting of halogen, --CN, --NO.sub.2,
C.sub.1-C.sub.4alkyl, haloC.sub.1-C.sub.4alkyl, --C(O)R.sub.6,
--C(O)R.sub.7, --C(O)NR.sub.7R.sub.7', --OR.sub.7, --OR.sub.6,
--NHR.sub.7, --NR.sub.7--(C.sub.1-C.sub.4alkyl),
--NR.sub.7-(haloC.sub.1-C.sub.4alkyl),
--NR.sub.7(CH.sub.2).sub.nC(O)R.sub.6, --NR.sub.6R.sub.7,
--NR.sub.7-heterocycloalkyl, wherein said heterocycloalkyl is
unsubstituted or substituted with 1-2 substituents selected from
R.sub.7, or --NR.sub.7SO.sub.2R.sub.7, or heterocycloalkyl that is
unsubstituted or substituted with 1-3 substituents selected from
halogen, C.sub.1-C.sub.4alkyl, haloC.sub.1-C.sub.4alkyl,
--(CH.sub.2).sub.nOH, --NR.sub.7R.sub.7', --OR.sub.7 or
--C(O)R.sub.7; wherein, R.sub.6 is --(CH.sub.2).sub.qOR.sub.7,
--(CH.sub.2).sub.qNR.sub.7R.sub.7',
--(CH.sub.2).sub.qNR.sub.7C(O)R.sub.7,
--(CH.sub.2).sub.qC(O)R.sub.7 or
--(CH.sub.2).sub.qC(O)NR.sub.7R.sub.7'; R.sub.7 and R.sub.7' are
each independently hydrogen, C.sub.1-C.sub.4alkyl,
C.sub.2-C.sub.6alkenyl, C.sub.2-C.sub.6alkynyl or
haloC.sub.1-C.sub.4alkyl, or R.sub.7, R.sub.7' and the nitrogen
atom attached thereto are cyclized together to form a
heterocycloalkyl that is unsubstituted or substituted with 1-3
substituents selected from halogen, C.sub.1-C.sub.4alkyl,
haloC.sub.1-C.sub.4alkyl, --(CH.sub.2).sub.nOH, --NR.sub.7R.sub.7',
--OR.sub.7 or --C(O)R.sub.7; m is 1, 2 or 3; n is 0, 1, 2, 3 or 4;
q is 0, 1, 2, 3 or 4.
The present invention provides a compound represented by the
general formula (I), which can inhibit one or more EGFR activating
or resistant mutations, such as L858R activating mutation, Exon 19
deletion activating mutation, and T790M resistant mutation.
Advantageously, the present compound can be useful in treating the
cancer patient who has been resistant to the existing therapy based
on the EGFR inhibitor.
The present invention provides a compound represented by the
general formula (I), which shows a higher inhibition to the
activating or resistant mutation-type EGFR than the wild-type EGFR.
Due to the reduced toxicity associated with the inhibition of the
wild-type EGFR, it is therefore expected that the compound of the
present invention is more useful as a therapeutic agent, in
particular for treating the cancer.
The present invention also provides a process for preparing the
compound represented by the general formula (I) of the present
invention.
The present invention also provides a pharmaceutical composition,
comprising the compound represented by the general formula (I) of
the present invention or a pharmaceutically acceptable salt
thereof, and a pharmaceutically acceptable carrier, excipient or
diluent.
The present invention also provides use of the compound represented
by the general formula (I) of the present invention or a
pharmaceutically acceptable salt thereof for treating an EGFR
activating or resistant mutation-mediated disease, in particular
cancer, in mammals, in particular human.
The present invention also provides use of the compound represented
by the general formula (I) of the present invention or a
pharmaceutically acceptable salt thereof in manufacture of a
medicament for treating an EGFR activating or resistant
mutation-mediated disease, in particular cancer, in mammals, in
particular human.
The present invention also provides a method for treating an EGFR
activating or resistant mutation-mediated disease, in particular
cancer, in mammals, in particular human, said method comprises
administrating to a patient the compound represented by the general
formula (I) or a pharmaceutically acceptable salt thereof, or the
pharmaceutical composition comprising a therapeutically effective
amount of the compound represented by the general formula (I) and a
pharmaceutically acceptable carrier, excipient or diluent.
The present invention also provides a method of selectively
inhibiting the EGFR activating or resistant mutation over the
wild-type EGFR (WT EGFR), said method comprises contacting a
biological sample with or administrating to a patient the compound
represented by the general formula (I) or a pharmaceutically
acceptable salt thereof or a pharmaceutical composition comprising
the same.
The cancer as mentioned in the present invention can be selected
from a group consisting of lung cancer, ovarian cancer, cervical
cancer, breast cancer, stomach cancer, colorectal cancer,
pancreatic cancer, glioma, glioblastoma, melanoma, prostate cancer,
leukemia, lymphoma, non-Hodgkin's lymphoma, hepatocytes cancer,
gastrointestinal stromal tumor (GIST), thyroid cancer,
cholangiocarcinoma, endometrial cancer, renal cancer, anaplastic
large cell lymphoma, acute myeloid leukemia (AML), multiple
myeloma, and mesothelioma.
In a preferable embodiment of the compound represented by the
general formula (I) or a pharmaceutically acceptable salt thereof
according to the present invention, Ring A is heteroaryl.
In a more preferable embodiment of the compound represented by the
general formula (I) or a pharmaceutically acceptable salt thereof
according to the present invention, Ring A is indolyl, indazolyl,
.[.pyrro[2,3-c]pyridinyl.]. .Iadd.pyrrolo[2,3-c]pyridinyl.Iaddend.,
.[.pyrro[3,2-c]pyridinyl.]. .Iadd.pyrrolo[3,2-c]pyridinyl.Iaddend.,
.[.pyrro[2,3-b]pyridinyl.]. .Iadd.pyrrolo[2,3-b]pyridinyl.Iaddend.,
.[.pyrro[3,2-b]pyridinyl.]. .Iadd.pyrrolo[3,2-b]pyridinyl.Iaddend.,
.[.pyrro[2,3-b]pyrazinyl.]. .Iadd.pyrrolo[2,3-b]pyrazinyl.Iaddend.,
indolin-2-onyl, pyridinyl, pyrazolyl or pyrimidinyl.
In a preferable embodiment of the compound represented by the
general formula (I) or a pharmaceutically acceptable salt thereof
according to the present invention, R.sub.1 is hydrogen, halogen or
haloC.sub.1-C.sub.4alkyl.
In a more preferable embodiment of the compound represented by the
general formula (I) or a pharmaceutically acceptable salt thereof
according to the present invention, R.sub.1 is hydrogen, chloro,
fluoro or trifluoromethyl.
In a preferable embodiment of the compound represented by the
general formula (I) or a pharmaceutically acceptable salt thereof
according to the present invention, R.sub.2 is C.sub.1-C.sub.4alkyl
or haloC.sub.1-C.sub.4alkyl, preferably C.sub.2-C.sub.4alkyl or
haloC.sub.2-C.sub.4alkyl, more preferably isopropyl or
trifluoroethyl.
In a preferable embodiment of the compound represented by the
general formula (I) or a pharmaceutically acceptable salt thereof
according to the present invention, R.sub.4 is
##STR00009## R.sub.7 and R.sub.7' are each independently hydrogen
or C.sub.1-C.sub.4alkyl.
In a more preferable embodiment of the compound represented by the
general formula (I) or a pharmaceutically acceptable salt thereof
according to the present invention, R.sub.4 is
##STR00010## R.sub.7 is hydrogen.
In a preferable embodiment of the compound represented by the
general formula (I) or a pharmaceutically acceptable salt thereof
according to the present invention, R.sub.3 is selected from a
group consisting of
halogen, --CN, --NO.sub.2, C.sub.1-C.sub.4alkyl,
haloC.sub.1-C.sub.4alkyl, --C(O)R.sub.7, --C(O)NR.sub.7R.sub.7',
--OR.sub.7, --NHR.sub.7, --NR.sub.7--(C.sub.1-C.sub.4alkyl),
--NR.sub.7(CH.sub.2).sub.nC(O)R.sub.6 or --NR.sub.6R.sub.7,
or heterocycloalkyl that is unsubstituted or substituted with 1-3
substituents selected from halogen, C.sub.1-C.sub.4alkyl,
haloC.sub.1-C.sub.4alkyl, --(CH.sub.2).sub.nOH, --NR.sub.7R.sub.7',
--OR.sub.7 or --C(O)R.sub.7;
wherein, R.sub.6 is --(CH.sub.2).sub.qOR.sub.7,
--(CH.sub.2).sub.qNR.sub.7R.sub.7', --(CH.sub.2).sub.qC(O)R.sub.7
or --(CH.sub.2).sub.qC(O)NR.sub.7R.sub.7';
R.sub.7 and R.sub.7' are each independently hydrogen,
C.sub.1-C.sub.4alkyl or haloC.sub.1-C.sub.4alkyl, or R.sub.7,
R.sub.7' and the nitrogen atom attached thereto are cyclized
together to form a heterocycloalkyl;
n is 0, 1, 2, 3 or 4;
q is 0, 1, 2, 3 or 4.
In a more preferable embodiment of the compound represented by the
general formula (I) or a pharmaceutically acceptable salt thereof
according to the present invention, R.sub.3 is --NR.sub.6R.sub.7,
wherein R.sub.6 is --(CH.sub.2).sub.qNR.sub.7R.sub.7', R.sub.7 and
R.sub.7' are each independently hydrogen or C.sub.1-C.sub.4alkyl, q
is 2.
In a more preferable embodiment of the compound represented by the
general formula (I) or a pharmaceutically acceptable salt thereof
according to the present invention, R.sub.3 is a heterocycloalkyl
substituted by one substituent selected from halogen,
C.sub.1-C.sub.4alkyl, haloC.sub.1-C.sub.4alkyl or
--NR.sub.7R.sub.7', R.sub.7 and R.sub.7' are each independently
hydrogen or C.sub.1-C.sub.4alkyl; more preferably, the
heterocycloalkyl is pyrrolidinyl.
In a preferable embodiment of the compound represented by the
general formula (I) or a pharmaceutically acceptable salt thereof
according to the present invention, each R.sub.5 is
.[.dependently.]. .Iadd.independently .Iaddend.halogen,
C.sub.1-C.sub.4alkyl, haloC.sub.1-C.sub.4alkyl, --OR.sub.7,
--NR.sub.7R.sub.7', --CN or --NO.sub.2, R.sub.7 and R.sub.7' are
each independently hydrogen or C.sub.1-C.sub.4alkyl, m is 1, 2 or
3.
In a more preferable embodiment of the compound represented by the
general formula (I) or a pharmaceutically acceptable salt thereof
according to the present invention, each R.sub.5 is
.[.dependently.]. .Iadd.independently .Iaddend.halogen,
C.sub.1-C.sub.4alkyl, --OR.sub.7 or --NR.sub.7R.sub.7', R.sub.7 and
R.sub.7' are each independently hydrogen or C.sub.1-C.sub.4alkyl, m
is 1, 2 or 3.
The specifically preferable compound represented by the general
formula (I) or a pharmaceutically acceptable salt thereof according
to the present invention includes:
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{5-chloro-[-
4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylamide;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[4-(1-meth-
yl-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylamide;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-(2,2,2-trifluoroethoxyl)-5-
-{[4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylamide;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-(2,2,2-trifluoroethoxyl)-5-
-{5-chloro-[4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}ac-
rylamide;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[-
4-(1-methyl-5-fluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acryl-
amide;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[4-(-
1-methyl-5,6-difluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acry-
lamide;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{5-c-
hloro-[4-(1-methyl-6-fluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-y-
l}acrylamide;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{5-chloro-[-
4-(1-methyl-5,6-difluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}a-
crylamide;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{-
5-chloro-[4-(1-methyl-5-fluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin--
3-yl}acrylamide;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{5-fluoro-[-
4-(1-methyl-5-fluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acryl-
amide;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{5-fl-
uoro-[4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylami-
de;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{5-fluor-
o-[4-(1-methyl-5,6-difluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-y-
l}acrylamide;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[4-(1-meth-
yl-6-fluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylamide;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-(2,2,2-trifluoroethoxy)-5--
{5-fluoro-[4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acr-
ylamide;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-(2,2,2-trifluoroet-
hoxyl)-5-{[4-(1-methyl-5-fluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-
-3-yl}acrylamide;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{5-chloro-[-
4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylamide
methanesulfonate;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{5-chloro-[-
4-(1-methyl-5,6-difluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}a-
crylamide methanesulfonate;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[4-(1-meth-
yl-5,6-difluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylamide
methanesulfonate;
.[.N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[5-chlo-
ro-4-(1-methyl-1H-pyrro[2,3-b]pyridin-3-yl)pyrimidin-2-yl]amino}pyridin-3--
yl}acrylamide.].
.Iadd.N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[5-c-
hloro-4-(1-methyl-1H-pyrrolo[2,3-b]pyridin-3-yl)pyrimidin-2-yl]amino}pyrid-
in-3-yl}acrylamide.Iaddend.;
.[.N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[5-chlo-
ro-4-(1-methyl-1H-pyrro[2,3-b]pyridin-5-yl)pyrimidin-2-yl]amino}pyridin-3--
yl}acrylamide.].
.Iadd.N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[5-c-
hloro-4-(1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)pyrimidin-2-yl]amino}pyrid-
in-3-yl}acrylamide.Iaddend.;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[5-chloro--
4-(1-methyl-1H-pyrazol-4-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylamide;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[5-chloro--
2'-methoxy-(4,5'-bipyrimidine)-2-yl]amino}pyridin-3-yl}acrylamide;
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[5-chloro--
2'-amino-(4,5'-bipyrimidine)-2-yl]amino}pyridin-3-yl}acrylamide.
The present invention also provides a process for preparing the
compound represented by the general formula (I), which comprises
the steps of:
##STR00011## ##STR00012## wherein ring A, R.sub.1, R.sub.2,
R.sub.3, R.sub.4, R.sub.5 and m are identical to those defined in
the above general formula (I); L represents a leaving group,
including hydrogen, halogen or
##STR00013## compounds (a) and (b) are used as starting material,
and subjected to substitution under a catalyst to produce an
Intermediate 2; the Intermediate 2 and an Intermediate 1 are
subjected to substitution or coupling reaction to produce a
compound (c), the nitro group of the compound (c) is reduced to
produce a compound (d), the compound (d) is acylated to produce a
compound (I); or the Intermediate 2 and an Intermediate 1' are
subjected to substitution or coupling reaction to directly produce
a compound (I).
In the process for preparing the compound of the general formula
(I), the catalyst for carrying out the substitution reaction of the
compounds (a) and (b) includes a Lewis acid such as AlCl.sub.3 or a
transition metal catalyst such as
bis(pinacolato)diboron/PdCl.sub.2(dppf), PdCl.sub.2(dppf); the
substitution or coupling reaction of Intermediate 2 and
Intermediate 1 can also be carried out under the catalysis of a
transition metal catalyst including but not limited to
Pd2(dba)3/xantphos; conventional reducing agents well known in the
art are used in the reduction of the nitro group, includes but is
not limited to iron powders, zinc powders, sodium sulfide,
H.sub.2/PtO.sub.2; the acylation of the compound (d) is carried out
with the corresponding acyl halide such as acyl chloride.
In an embodiment of preparing the compound represented by the
general formula (I) according to the present invention, if
Intermediate 2 is Intermediate 2j, the preparation process is as
follows,
##STR00014##
In the process for preparing the compound represented by the
general formula (I) according to the present invention, the
preparation process for Intermediate 1 and Intermediate 1'
comprises the steps of,
##STR00015## wherein, R.sub.2, R.sub.3 and R.sub.4 are defined as
in the above general formula (I); 2,6-dichloro-3-nitropyridine is
used as starting material, and subjected to etherification to
produce a compound (e), the nitro group of the compound (e) is
reduced to produce a compound (f), the compound (f) is then
subjected to a reaction to produce a compound (g), the compound (g)
is subjected to nitration to produce a compound (h), the compound
(h) and R.sub.3H are subjected to substitution to produce a
compound (i), and the compound (i) is then deprotected to produce
the Intermediate 1; the compound (i) is subjected to Boc-protection
to produce a compound (j), the compound (j) is then subjected to
acetyl-deprotection to produce a compound (k), the nitro group of
the compound (k) is reduced to produce a compound (l), the compound
(l) is subjected to acylation to produce a compound (m), and
finally the compound (m) is subjected to Boc-deprotection to
produce the Intermediate 1'.
In the process for preparing Intermediate 1 and Intermediate 1'
according to the present invention, the etherification reaction is
carried out in presence of a strong base which includes, but is not
limited to sodium hydride, potassium hydride, sodium hydroxide,
potassium hydroxide, sodium ethoxide, and sodium methoxide;
conventional reducing agents well known in the art are used in the
reduction of the nitro group, includes but is not limited to iron
powders, zinc powders, sodium sulfide, H.sub.2/PtO.sub.2;
protection and deprotection are the conventional method well known
in the art, and are carried out in an appropriate acidic or basic
condition.
In the above preparation processes, the used abbreviations for the
agents have the following meanings:
TABLE-US-00001 AlCl.sub.3 Aluminium chloride Bis(pinacolato)diboron
Bis(pinacolato)diboron PdCl.sub.2(dppf)
[1,1'-Bis(diphenylphosphino)ferrocene]dichloro- palladium
Pd.sub.2(dba).sub.3 Tris(dibenzylideneacetone)dipalladium Xantphos
4,5-Bis(diphenylphosphino)-9,9-dimethylxanthene PtO.sub.2 Platinum
dioxide NaH Sodium hydride THF tetrahydrofuran p-TsCl
p-Toluenesulfonyl chloride KOAc Potassium acetate Dioxane Dioxane
Na.sub.2CO.sub.3 Sodium carbonate TBAF Tetrabutylammonium fluoride
MeI Methyl iodide DMF N,N-dimethylformamide
In the present invention, the term "halogen" means fluoro, chloro,
bromo iodo and the like, preferably fluoro, chloro and bromo, and
more preferably chloro.
In the present invention, the term "C.sub.1-C.sub.4alkyl" means
methyl, ethyl, propyl, isopropyl, butyl, iso-butyl, sec-butyl or
tert-butyl, the term "C.sub.2-C.sub.4alkyl" means ethyl, propyl,
isopropyl, butyl, iso-butyl, sec-butyl or tert-butyl, preferably
ethyl, propyl, isopropyl or butyl, more preferably isopropyl.
In the present invention, the term "haloC.sub.1-C.sub.4alkyl" means
the C.sub.1-C.sub.4alkyl, as defined herein, which is substituted
with one or more halogen atoms, preferably 1-5 halogen atoms,
including but not limited to, trifluoromethyl, trifluoroethyl,
difluoromethyl, 1-chloro-2-fluoroethyl and the like. The term
"haloC.sub.2-C.sub.4alkyl" includes but is not limited to
trifluoroethyl, difluoromethyl, 1-chloro-2-fluoroethyl and the
like, preferably trifluoroethyl.
In the present invention, the term "alkenyl" means a mono-valent
group derived from a hydrocarbon group, the term
"C.sub.2-C.sub.6alkenyl" means an alkenyl group containing 2 to 6
carbon atoms and at least containing one C--C double bond,
including but not limited to, ethenyl, propenyl, butenyl,
2-methyl-2-butenyl, 2-methyl-2-pentenyl and the like.
In the present invention, the term "alkynyl" means a mono-valent
group derived from a hydrocarbon group, the term
"C.sub.2-C.sub.6alkynyl" means an alkynyl group containing 2 to 6
carbon atoms and at least containing one C--C triple bond,
including but not limited to, ethynyl, propynyl, 1-butynyl,
2-butynyl and the like.
In the present invention, the term "cycloalkyl" means a mono-valent
group derived from monocyclic or polycyclic, saturated or partially
unsaturated aliphatic carbocyclic compounds, the term
"C.sub.3-C.sub.8-cycloalkyl" includes but is not limited to
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,
cyclooctyl, cyclooctenyl, cyclopropenyl, cyclobutenyl,
cyclopentenyl, cyclohexenyl, cycloheptenyl, and the term
"C.sub.9-C.sub.12-" includes but is not limited to
bicyclo[2.2.1]heptyl, bicyclo[2.2.1]octyl and the like.
In the present invention, the term "heterocycloalkyl" means a
monovalent monocyclic group, which is saturated or partially
unsaturated (but not aromatic) and contains 3-8 ring members,
preferably 4-7 ring members, or a monovalent fused bicyclic group,
which is saturated or partially unsaturated (but not aromatic) and
contains 5-12 ring members, preferably 7-10 ring members, wherein
1-4 ring heteroatom(s) is/are selected from a group consisting of
O, S and N, and the remaining ring atoms are carbon. Said
heterocycloalkyl includes but is not limited to azetidinyl,
oxetanyl, pyrrolidinyl, piperidyl, morpholinyl, piperazinyl,
tetrahydropyranyl, pyrazolidinyl, pyrazolinyl, imidazolinyl,
imidazolidinyl, [1,3]dioxolane (dioxolane), dihydropyridinyl,
tetrahydropyridinyl, hexahydropyridinyl, oxazolinyl, oxazolidinyl,
iso-oxazolidinyl, thiazolinyl, thiazolidinyl, tetrahydrothiazolyl,
iso-tetrahydrothiazolyl, octahydroindolyl, octahydroisoindolyl,
tetrahydrofuryl and the like, preferably azetidinyl, oxetanyl,
pyrrolidinyl, piperidyl, morpholinyl or piperazinyl.
In the present invention, the term "aryl" means an aromatic cyclic
hydrocarbyl, which is a fused or non-fused carbonaceous ring system
containing one or more aromatic rings, and includes but is not
limited to phenyl, naphthyl, tetrahydronaphthyl, indanyl, indenyl
and the like, preferably an aryl containing 6-14 carbon atoms, more
preferably an aryl containing 6-10 carbon atoms, such as phenyl and
naphthyl, more preferably phenyl.
In the present invention, the term "heteroaryl" means 5-6 membered
monocyclic heteroaryl containing 1-4 heteroatoms selected from N, S
or O, or bicyclic heteroaryl formed by fusing said 5-6 membered
monocyclic heteroaryl with a benzene ring, pyridine ring or pyrrole
ring, said heteroaryl can be partially saturated. Said heteroaryl
includes but is not limited to furyl, thienyl, pyrrolyl,
imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl,
isoxazolyl, triazolyl, tetrazolyl, thiadiazolyl, pyridinyl,
pyrimidinyl, pyridazinyl, pyrazinyl, benzofuranyl, benzothienyl,
benzothiadiazolyl, benzothiazolyl, benzimidazolyl, indolyl,
isoindolyl, indazolyl, quinolyl, isoquinolyl, quinazolinyl,
1,2,3,4-tetrahydroisoquinolyl, .[.pyrro[2,3-c]pyridinyl.].
.Iadd.pyrrolo[2,3-c]pyridinyl.Iaddend., .[.pyrro[3,2-c]pyridinyl.].
.Iadd.pyrrolo[3,2-c]pyridinyl.Iaddend., .[.pyrro[2,3-b]pyridinyl.].
.Iadd.pyrrolo[2,3-b]pyridinyl.Iaddend., .[.pyrro[3,2-b]pyridinyl.].
.Iadd.pyrrolo[3,2-b]pyridinyl.Iaddend., .[.pyrro[2,3-b]pyrazinyl.].
.Iadd.pyrrolo[2,3-b]pyrazinyl.Iaddend., indolin-2-onyl, preferably
indolyl, indazolyl, .[.pyrro[2,3-c]pyridinyl.].
.Iadd.pyrrolo[2,3-c]pyridinyl.Iaddend., .[.pyrro[3,2-c]pyridinyl.].
.Iadd.pyrrolo[3,2-c]pyridinyl.Iaddend., .[.pyrro[2,3-b]pyridinyl.].
.Iadd.pyrrolo[2,3-b]pyridinyl.Iaddend., .[.pyrro[3,2-b]pyridinyl.].
.Iadd.pyrrolo[3,2-b]pyridinyl.Iaddend., .[.pyrro[2,3-b]pyrazinyl.].
.Iadd.pyrrolo[2,3-b]pyrazinyl.Iaddend., indolin-2-onyl, pyridinyl,
pyrazolyl or pyrimidinyl, imidazolyl, pyrazinyl, benzimidazolyl,
indolyl, isoindolyl or 1,2,3,4-tetrahydroisoquinolyl, more
preferably indolyl, indazolyl, .[.pyrro[2,3-c]pyridinyl.].
.Iadd.pyrrolo[2,3-c]pyridinyl.Iaddend., .[.pyrro[3,2-c]pyridinyl.].
.Iadd.pyrrolo[3,2-c]pyridinyl.Iaddend., .[.pyrro[2,3-b]pyridinyl.].
.Iadd.pyrrolo[2,3-b]pyridinyl.Iaddend., .[.pyrro[3,2-b]pyridinyl.].
.Iadd.pyrrolo[3,2-b]pyridinyl.Iaddend., .[.pyrro[2,3-b]pyrazinyl.].
.Iadd.pyrrolo[2,3-b]pyrazinyl.Iaddend., indolin-2-onyl, pyridinyl,
pyrazolyl or pyrimidinyl.
The present invention also includes the pharmaceutically acceptable
salt of the compound represented by formula (I). The term
"pharmaceutically acceptable salt" means relatively nontoxic acid
addition salts or base addition salts of the compound of the
present invention. Said acid addition salts are the salts formed
between the compound represented by formula (I) of the present
invention and suitable inorganic acids or organic acids. Said salts
may be prepared during the final separation and purification
processes of the compounds, or may be prepared through the reaction
of purified compound represented by formula (I) in the form of free
base thereof and suitable organic acids or inorganic acids.
Representative acid addition salts includes hydrobromic acid salt,
hydrochloric acid salt, sulfate, bisulfate, sulfite, acetate,
oxalate, valerate, oleate, palmate, stearate, laurate, borate,
benzoate, lactate, phosphate, hydrogen phosphate, carbonate,
bicarbonate, toluate, citrate, maleate, fumarate, succinate,
tartrate, benzoate, mesylate, p-tosylate, glyconate, lactobionate
and laurylsulfonate and the like. Said base addition salts are the
salts formed between the compound represented by formula (I) and
suitable inorganic bases or organic bases, including such as the
salts formed with alkali metals, alkaline earth metals, quaternary
ammonium cations, such as sodium salts, lithium salts, potassium
salts, calcium salts, magnesium salts, tetramethylammonium salts,
tetraethylammonium salt and the like; amine salts, including the
salts formed with ammonia (NH.sub.3), primary amines, secondary
amines or tertiary amines, such as: methylamine salts,
dimethylamine salts, trimethylamine salts, triethylamine salts,
ethylamine salts and the like.
The compound of the present invention or a pharmaceutically
acceptable salt thereof can be administered to mammals, such as
human, and administrated orally, rectally, parenterally
(intravenously, intramuscularly or subcutaneously), topically (such
as in the form of powders, ointments or drops), or
intratumorally.
The administration dosage of the compound of the present invention
can be about 0.05-50 mg/kg body weight/day, e.g. 0.1-45 mg/kg body
weight/day, 0.5-35 mg/kg body weight/day.
The compound of the present invention or a pharmaceutically
acceptable salt thereof can be formulated into the solid dosage
forms for oral administration, which includes but is not limited to
capsules, tablets, pills, powders and granules and the like. In
these solid dosage forms, the compounds represented by formula (I)
of the present invention as active ingredients are admixed with at
least one conventional inert excipients (or carriers), such as
sodium citrate or dicalcium phosphate, or admixed with the
following ingredients: (1) fillers or extenders, such as, starch,
lactose, sucrose, glucose, mannitol and silicic acid and the like;
(2) adhesives, such as, hydroxymethylcellulose, alginate, gelatin,
polyvinyl pyrrolidine, sucrose and acacia and the like; (3)
humectants, such as, glycerol and the like; (4) disintegrating
agents, such as, agar, calcium carbonate, potato starch or tapioca,
alginic acid, certain composite silicate and sodium carbonate and
the like; (5) retarding solvents, such as paraffin wax and the
like; (6) absorption accelerators, such as, quaternary ammonium
compounds and the like; (7) moistening agents, such as cetanol and
glyceryl monostearate and the like; (8) absorbents, such as, kaolin
and the like; and (9) lubricants, such as, talc, calcium stearate,
magnesium stearate, solid polyethylene glycol, sodium dodecyl
sulphate and the like, or mixtures thereof. Capsules, tablets and
pills may also comprise buffers.
Said solid dosage forms such as tablets, sugar pills, capsules,
pills and granules can also by coated or microencapsulated by
coatings and shell materials such as enteric coatings and other
materials well known in the art. They may comprise opacifying
agents, and the release of active ingredients in these compositions
may be carried out in a certain portion of digestive tube in a
retarded manner. The examples for embedding components that may be
adopted are polymers and waxes. If necessary, active ingredients
can also be formulated into the form of microcapsules with one or
more of the above excipients.
The compound of the present invention or a pharmaceutically
acceptable salt thereof can be formulated into liquid dosage forms
for oral administration, including but not limited to
pharmaceutically acceptable emulsions, solutions, suspensions,
syrups and tinctures and the like. Besides the compounds
represented by formula (I) or a pharmaceutically acceptable salt
thereof as active ingredients, the liquid dosage forms may comprise
inert diluents customarily used in the art, such as water and other
solvents, solubilizers and emulsifiers, such as, ethanol, isopropyl
alcohol, ethyl carbonate, ethyl acetate, propylene glycol,
1,3-butanediol, dimethyl formamide, and oils, especially cottonseed
oil, peanut oil, corn germ oil, olive oil, castor oil and sesame
oil and the like or mixtures of these materials and the like.
Besides these inert diluents, the liquid dosage forms of the
present invention may also comprise conventional auxiliaries, such
as moistening agents, emulsifiers and suspending agents, sweeting
agents, flavoring agents and fragrances and the like.
Said suspending agents includes, such as, ethoxylated isostearyl
alcohol, polyoxyethylene sorbitol and sorbitan ester,
microcrystalline cellulose, aluminium methoxide and agar and the
like or mixtures of these materials.
The compound of the present invention or a pharmaceutically
acceptable salt thereof can be formulated into dosage forms for
parenteral injection, including but not limited to physiologically
acceptable sterile aqueous or anhydrous solutions, dispersions,
suspensions or emulsions, and sterile powder for re-dissolving into
sterile injectable solutions or dispersions. Suitable carriers,
diluents, solvents or excipients include water, ethanol, polyhydric
alcohol and suitable mixtures thereof.
The compound of the present invention or a pharmaceutically
acceptable salt thereof can also be formulated into dosage forms
for topical administration, including but not limited to ointments,
powders, suppositories, drops, propellants and inhalants and the
like. The compounds represented by formula (I) of the present
invention or a pharmaceutically acceptable salt thereof as active
ingredients are admixed together with physiologically acceptable
carriers and optional preservatives, buffers, or if necessary,
propellants, under sterile condition.
The present invention also provides a pharmaceutical composition
containing the compound represented by formula (I) of the present
invention or a pharmaceutically acceptable salt thereof as active
ingredients, and pharmaceutically acceptable carriers, excipients
or diluents. When preparing the pharmaceutical composition, the
compound represented by formula (I) of the present invention or a
pharmaceutically acceptable salt thereof is generally admixed with
pharmaceutically acceptable carriers, excipients or diluents. The
content of the compound of the general formula (I) or a
pharmaceutically acceptable salt thereof can be 0.01-1000 mg, for
example 0.05-800 mg, 0.1-500 mg, 0.01-300 mg, 0.01-200 mg, 0.05-150
mg, 0.05-50 mg and the like.
By conventional preparation methods, the composition of the present
invention may be formulated into conventional pharmaceutical
preparations, such as tablets, pills, capsules, powder, granules,
emulsions, suspensions, dispersions, solutions, syrups, elixirs,
ointments, drops, suppositories, inhalants, propellants and the
like.
The compound of the present invention or a pharmaceutically
acceptable salt thereof may be administered alone or in combination
with other pharmaceutically acceptable therapeutic agents,
especially with other anti-tumor drugs. The therapeutic agents
include but are not limited to anti-tumor drugs which exert an
influence on the chemical structure of DNA, such as Cisplatin,
anti-tumor drugs which affect the synthesis of nucleic acid, such
as Methotrexate (MTX), 5-Fluorouracil (5FU) and the like,
anti-tumor drugs which affect the transcription of nucleic acid,
such as Adriamycin, Epirubicin, Aclacinomycin, Mitramycin and the
like, anti-tumor drugs which exert an influence on synthesis of
tubulin, such as Paclitaxel, Vinorelbine and the like, aromatase
inhibitors such as Aminoglutethimide, Lentaron, Letrozole,
Anastrozole and the like, inhibitors of the cell signal pathway
such as epidermal growth factor receptor inhibitors Imatinib,
Gefitinib, Erlotinib, and the like. Each therapeutic agent to be
combined can be administered simultaneously or sequentially, and
can be administered either in a unitary formulation or in separate
formulations. Such combination includes not only the combination of
the compound of the present invention with another active
ingredient but also the combination of the compound of the present
invention with two or more other active ingredients.
It is proved by the cell experiments, i.e., in vitro proliferation
inhibition experiments on the activating mutation, i.e., Exon 19
deletion activating mutation tumor cells, such as HCC827 cell,
resistant tumor cells such as H1975 and the wild-type EGFR human
skin cancer cell A431 that, the compound of the present invention
has a good proliferation inhibition effect on the activating
mutation or resistant mutation tumor cells and a weak proliferation
inhibition effect on the wild-type EGFR cancer cells, and has a
good selectivity. It is proved by the animal experiment, i.e., the
experiment of inhibiting the growth of subcutaneously transplanted
tumors of human lung cancer H1975-bearing nude mice that, the
compound of the present invention has a good inhibition effect on
the growth of the transplanted tumor and a good safety. The
compound of the present invention can be used as the medicament for
treating a disease or condition mediated by the activity of EGFR
activating or resistant mutation, in particular tumor, e.g. cancer.
Said cancer includes but is not limited to, e.g. lung cancer,
ovarian cancer, cervical cancer, breast cancer, stomach cancer,
colorectal cancer, pancreatic cancer, glioma, glioblastoma,
melanoma, prostate cancer, leukemia, lymphoma, non-Hodgkin's
lymphoma, hepatocytes cancer, gastrointestinal stromal tumor
(GIST), thyroid cancer, cholangiocarcinoma, endometrial cancer,
renal cancer, anaplastic large cell lymphoma, acute myeloid
leukemia (AML), multiple myeloma, mesothelioma, in particular a
type of tumor wherein threonine at position 790 of the epidermal
growth factor receptor is mutated into methionine (EGFR T790M). For
example, the compound of the present invention can be used as
medicament for treating the non-small cell cancer (EGFR T790M). It
can be used to overcome the resistency problem caused by EGFR T790M
after Gefitinib and Erlotinib are clinically used. Due to the
reduced toxicity associated with the inhibition of the wild-type
EGFR, it is therefore expected that the compound of the present
invention will produce a relatively small toxic and side-effect
upon being applied to the cancer treatment.
The pharmacodynamic action of the compound of the present invention
in terms of inhibiting the proliferation of cancer cells may be
assayed by conventional methods. One preferable evaluation method
of which is Sulforhodamine B (SRB) protein staining method, which
calculates the inhibition ratio of a drug against the proliferation
of cancer cells by measuring the change in optical absorption value
generated after the drug has acted on the cancer cells. Inhibition
ratio (%)=[(blank control OD-inhibitor OD)/blank control
OD].times.100%
Blank control OD: the OD value of the well of normally growed cells
without the action of a drug.
Inhibitor OD: the OD value of the well of cells with the action of
the added compounds to be screened.
The median inhibitory concentration (IC.sub.50) value is obtained
by the software GraphPad Prism 5.0 by the 4-parameter logistic
curve fit calculation. Each experiment is repeated three times, and
the average IC.sub.50 value for three experiments is used as the
final index for the inhibitory ability.
The pharmacodynamic action of the compound of the present invention
in terms of inhibiting the growth of transplanted tumors in animal
may be assayed by conventional methods. One preferable evaluation
method of which is the inhibitory effect on the growth of
subcutaneously transplanted tumors of human lung cancer
H1975-bearing nude mice. The experimental method is as follows:
human lung cancer H1975 cell strain (5.times.10.sup.6/each mouse)
is inoculated to nude mice subcutaneously at the right side of the
back thereof. After the tumors grow to 100-150 mm.sup.3 on average,
the animals are divided into groups randomly according to the tumor
size and the animal weight. The test compounds are administered by
intragastric administration in a certain dosages, and solvent
control groups are administered with equal amount of solvent by
intragastric administration, wherein the administration is
performed once per day for a continuous period of 12 days. During
the entire experimental process, the animal weight and the tumor
size are measured twice per week, so as to observe whether or not
the toxic reaction occurs. The tumor volume is calculated as
follows: Tumor volume (mm.sup.3)=0.5.times.(Tumor major
diameter.times.Tumor minor diameter.sup.2)
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 is the tumor volume curve for subcutaneously transplanted
tumors of human lung cancer H1975-bearing nude mice at the
administration dosage of 25 mg/kg of the compound of Example 3 and
AZD9291.
FIG. 2 is the body weight curve for human lung cancer H1975-bearing
nude mice at the administration dosage of 25 mg/kg of the compound
of Example 3 and AZD9291.
The present invention will be further illustrated hereinafter in
connection with specific Examples. It should be understood that
these Examples are only used to illustrate the present invention by
the way of examples without limiting the scope thereof. In the
following Examples, the experimental methods without specifying
conditions are generally performed according to conventional
conditions or based on the conditions recommended by the
manufacturer. The parts and percentages are the parts and
percentages by weight respectively, unless otherwise specified.
DETAILED DESCRIPTION OF THE INVENTION
I. Preparation Examples of the Compounds of the Present
Invention
Intermediate 1a:
N.sup.2-methyl-N.sup.2-[2-(dimethylamino)ethyl]-6-methoxy-3-nitropyridin--
2,5-diamine hydrochloride
##STR00016##
Step 1: Synthesis of 6-chloro-2-methoxy-3-nitropyridine
##STR00017##
To a 250 mL three-necked flask were added
2,6-dichloro-3-nitropyridine (11.58 g, 60 mmol), 150 ml
tetrahydrofuran and methanol (1.92 g, 60 mmol). The mixture was
cooled to 0.degree. C. To the mixture was added in batch 60% sodium
hydride (2.4 g, 60 mmol). The resulting mixture was stirred at
0.degree. C. for 1 hour, warmed up slowly to room temperature, and
continued to stir for 1 hour. To the reaction mixture was added 100
ml ethyl acetate. The reaction mixture was washed successively with
water (50 ml.times.2) and saturated brine (50 ml). The organic
phase was dried with anhydrous sodium sulfate, filtered, evaporated
under a reduced pressure to remove the solvent, purified by silica
gel column chromatography (petroleum ether:ethyl acetate=30:1) to
produce 7.3 g of a product with a yield of 64%.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.29 (d, J=8.3 Hz, 1H),
7.07 (d, J=8.3 Hz, 1H), 4.15 (s, 3H).
Step 2: Synthesis of 6-chloro-2-methoxypyridin-3-amine
##STR00018##
To a 100 mL single-necked flask were added
6-chloro-2-methoxy-3-nitropyridine (2.0 g, 10.6 mmol), ammonia
chloride (2.8 g, 53.0 mmol) and 80 ml of a mixed solvent of ethanol
and water (volume ratio=3:1). To the mixture was added in batch a
reduced iron powders (3.0 g, 53.0 mmol). The mixture was stirred at
80.degree. C. for 1.5 hours. The reaction mixture was cooled to
room temperature, and filtered through diatomite. 150 ml ethyl
acetate and 120 ml saturated sodium chloride were added to the
filtrate. An organic layer was separated and dried with anhydrous
sodium sulfate, and filtered. The filtrate was evaporated to
dryness under a reduced pressure to produce a brown solid (1.6 g)
with a yield of 95%. MS m/z: 159 [M+1].
Step 3: Synthesis of
N-(6-chloro-2-methoxypyridin-3-yl)acetamide
##STR00019##
To a 250 mL single-necked flask were added
6-chloro-2-methoxypyridin-3-amine (1.6 g, 10.1 mmol),
diisopropylethylamine (2.6 ml, 15.1 mmol) and 100 ml
dichloromethane. The mixture was cooled to 5.degree. C. in an ice
bath. Acetyl chloride (0.86 ml, 12.1 mmol) was added. The reaction
continued for 1.25 hours. The reaction mixture was washed
successively with 80 ml water, 80 ml 1N hydrochloric acid and 80 ml
saturated sodium chloride solution, dried with anhydrous sodium
sulfate, filtered, and evaporated to dryness under a reduced
pressure to produce 1.9 g of a brown solid with a yield of 94%. MS
m/z: 201 [M+1].
Step 4: Synthesis of
N-(6-chloro-2-methoxy-5-nitropyridin-3-yl)acetamide
##STR00020##
To a 100 mL single-necked flask were added
N-(6-chloro-2-methoxypyridin-3-yl)acetamide (1.9 g, 9.47 mmol) and
20 ml trifluoroacetic anhydride. The mixture was cooled in an
ice-salt bath to -10.degree. C. Fuming nitric acid (0.4 ml, 9.47
mmol) was dropwisely added while the temperature was controlled to
below -5.degree. C. After the completion of dropwise addition, the
reaction continued in an ice-salt bath for 1.25 hours. The reaction
mixture was slowly added to crushed ice. A solid precipitated and
was filtered. The resulting crude product was dried at 60.degree.
C., and added to ethyl acetate to form a slurry. 1.5 g of an beige
solid was obtained with a yield of 65%. MS m/z: 244 [M-1].
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.90 (s, 1H), 9.17 (s, 1H),
4.06 (s, 3H), 2.17 (s, 3H).
Step 5: Synthesis of
N-{6-{[2-(dimethylamino)ethyl](methyl)amino}-2-methoxy-5-nitropyridin-3-y-
l}acetamide hydrochloride
##STR00021##
To a 100 mL single-necked flask were added
N-(6-chloro-2-methoxy-5-nitropyridin-3-yl)acetamide 1.0 g, 4.1
mmol), 30 ml acetonitrile and N,N,N'-trimethylethylenediamine (0.6
g, 6.1 mmol). The mixture was reacted at 80.degree. C. for 3 hours.
The reaction mixture was concentrated under a reduced pressure to
about 1/3 of the original volume. 50 ml ethyl acetate was added.
The mixture was stirred for several minutes, a solid precipitated
and was filtered to produce 1.1 g of an beige solid with a yield of
87%.
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 11.13 (s, 1H), 9.53 (s, 1H),
8.73 (s, 1H), 4.05 (s, 5H), 3.41 3.36 (m, 2H), 2.83 (s, 3H), 2.80
(s, 6H), 2.07 (s, 3H).
Step 6: Synthesis of
N.sup.2-methyl-N.sup.2-[2-(dimethylamino)ethyl]-6-methoxy-3-nitropyridin--
2,5-diamine hydrochloride
##STR00022##
To a 50 mL single-necked flask were added
N-{6-{[2-(dimethylamino)ethyl](methyl)amino}-2-methoxy-5-nitropyridin-3-y-
l}acetamide (600 mg, 1.93 mmol), 15 ml methanol and 0.3 ml
concentrated hydrochloric acid. The mixture was reacted at
60.degree. C. overnight. The reaction mixture was evaporated to
dryness under a reduced pressure. 100 ml dichloromethane and 80 ml
saturated sodium bicarbonate were added. The resulting mixture was
stirred until no bubble produced. An organic layer was separated
and dried with anhydrous sodium sulfate, filtered, and concentrated
under a reduced pressure. The residue was purified by silica gel
column chromatography (dichloromethane:methanol=10:1) to produce
400 mg of a brown solid. MS m/z: 270 [M+1].
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 11.20 (s, 1H), 8.16 (s, 1H),
4.06-4.02 (m, 5H), 3.38 (br s, 2H), 2.83 (s, 3H), 2.80 (s, 3H),
2.79 (s, 3H).
Intermediate 1b: Synthesis of
N.sup.2-methyl-N.sup.2-[2-(dimethylamino)ethyl]-6-isopropyloxy-3-nitropyr-
idin-2,5-diamine
##STR00023##
Step 1: Synthesis of 6-chloro-2-isopropyloxy-3-nitropyridine
##STR00024##
The compound was synthesized in the same manner as those in Step 1
of Intermediate 1a.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.22 (d, J=8.3 Hz, 1H),
6.98 (d, J=8.3 Hz, 1H), 5.50 (hept, J=6.2 Hz, 1H), 1.43 (d, J=6.2
Hz, 6H).
Step 2: Synthesis of 6-chloro-2-isopropyloxypyridin-3-amine
##STR00025##
The compound was synthesized in the same manner as those in Step 2
of Intermediate 1a with a yield of 74%. MS m/z: 187 [M+1], 189.
Step 3: Synthesis of
N-(6-chloro-2-isopropyloxypyridin-3-yl)acetamide
##STR00026##
The compound was synthesized in the same manner as those in Step 3
of Intermediate 1a with a yield of 83%. MS m/z: 229 [M+1], 231.
Step 4: Synthesis of
N-(6-chloro-2-isopropyloxy-5-nitropyridin-3-yl)acetamide
##STR00027##
The compound was synthesized in the same manner as those in Step 4
of Intermediate 1a with a yield of 33%. MS m/z: 272 [M-1].
Step 5: Synthesis of
N-{6-{[2-(dimethylamino)ethyl](methyl)amino}-2-isopropyloxy-5-nitropyridi-
n-3-yl}acetamide
##STR00028##
To a 500 mL single-necked flask were added
N-(6-chloro-2-isopropyloxy-5-nitropyridin-3-yl)acetamide (15 g,
54.8 mmol), 150 ml acetonitrile, N,N,N'-trimethylethylenediamine
(7.28 g, 71.3 mmol) and potassium carbonate (15.15 g, 110 mmol).
The mixture was reacted at 80.degree. C. overnight. The reaction
mixture was cooled to room temperature, and filtered. The filtrate
was evaporated to dryness under a reduced pressure to produce 18.6
g of a product with a yield of 100%.
MS m/z: 340 [M+1].
Step 6: Synthesis of
N.sup.2-methyl-N.sup.2-[2-(dimethylamino)ethyl]-6-isopropyloxy-3-nitropyr-
idin-2,5-diamine
##STR00029##
The compound was synthesized in the same manner as those in Step 6
of Intermediate 1a with a yield of 38%. MS m/z: 298 [M+1].
Intermediate 1c:
N.sup.2-methyl-N.sup.2-[2-(dimethylamino)ethyl]-6-(2,2,2-trifluoroethoxyl-
)-3-nitropyridin-2,5-diamine
##STR00030##
Step 1: Synthesis of
6-chloro-2-(2,2,2-trifluoroethoxyl)-3-nitropyridine
##STR00031##
The compound was synthesized in the same manner as those in Step 1
of Intermediate 1a with a yield of 80%.
Step 2: Synthesis of
6-chloro-2-(2,2,2-trifluoroethoxyl)pyridin-3-amine
##STR00032##
The compound was synthesized in the same manner as those in Step 2
of Intermediate 1a with a yield of 83%.
Step 3: Synthesis of
N-[6-chloro-2-(2,2,2-trifluoroethoxyl)pyridin-3-yl]acetamide
##STR00033##
The compound was synthesized in the same manner as those in Step 3
of Intermediate 1a with a yield of 71%. MS m/z: 269 [M+1], 271.
Step 4: Synthesis of
N-[6-chloro-2-(2,2,2-trifluoroethoxyl)-5-nitropyridin-3-yl]acetamide
##STR00034##
The compound was synthesized in the same manner as those in Step 4
of Intermediate 1a with a yield of 53%. MS m/z: 314 [M+1], 316.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.37 (s, 1H), 7.63 (s,
1H), 4.93 (q, J=8.2 Hz, 2H), 2.30 (s, 3H).
Step 5: Synthesis of
N-{6-{[2-(dimethylamino)ethyl](methyl)amino}-2-(2,2,2-trifluoroethoxyl)-5-
-nitropyridin-3-yl}acetamide
##STR00035##
To a 25 mL single-necked flask were added
N-[6-chloro-2-(2,2,2-trifluoroethoxyl)]-5-nitropyridin-3-yl)acetamide
(626 mg, 2 mmol), 10 ml acetonitrile,
N,N,N'-trimethylethylenediamine (224 mg, 2.2 mmol) and potassium
carbonate (138 mg, 4 mmol). The mixture was stirred at room
temperature overnight. To the reaction mixture was added 100 ml
ethyl acetate. The resulting mixture was washed with 20 ml water,
dried with anhydrous sodium sulfate, and evaporated under a reduced
pressure to remove the solvent to produce 710 mg of a product with
a yield of 94%. MS m/z: 380 [M+1].
Step 6: Synthesis of
N.sup.2-methyl-N.sup.2-[2-(dimethylamino)ethyl]-6-(2,2,2-trifluoroethoxyl-
)-3-nitropyridin-2,5-diamine
##STR00036##
The compound was synthesized in the same manner as those in Step 6
of Intermediate 1a with a yield of 100%. MS m/z: 338 [M+1].
Intermediate 1d: tert-butyl
{5-acrylamide-6-{[2-(dimethylamino)ethyl](methyl)amino}-2-isopropyloxypyr-
idin-3-yl}carbamate
##STR00037##
Step 1: Synthesis of
N-tert-butoxycarbonyl-N-{6-{[2-(dimethylamino)ethyl](methyl)amino}-2-isop-
ropyloxy-5-nitropyridin-3-yl}acetamide
##STR00038##
To a 500 mL single-necked flask were added
N-{6-{[2-(dimethylamino)ethyl](methyl)amino}-2-isopropyloxy-5-nitropyridi-
n-3-yl}acetamide (18.6 g, 54.8 mmol), 4-dimethylaminopyridine (0.67
g, 5.48 mmol), 150 ml acetonitrile and di-tert-butyl dicarbonate
(59.8 g, 274 mmol). The mixture was reacted at 80.degree. C. for
2.5 hours. The reaction mixture was cooled to room temperature, was
evaporated to dryness under a reduced pressure, and purified by
silica gel column chromatography (dichloromethane methanol=10:1) to
produce 24 g of a product with a yield of 100%.
Step 2: Synthesis of tert-butyl
{6-({[2-(dimethylamino)ethyl](methyl)amino}-2-isopropyloxy-5-nitropyridin-
-3-yl}carbamate
##STR00039##
To a 500 mL single-necked flask were added
N-tert-butoxycarbonyl-N-{6-{[2-(dimethylamino)ethyl](methyl)amino}-2-isop-
ropyloxy-5-nitropyridin-3-yl}acetamide (24 g, 54.6 mmol) and 240 ml
methanol. The mixture was cooled to 0.degree. C. Sodium methoxide
(2.95 g, 54.6 mmol) was added. The mixture was slowly warmed up to
room temperature and reacted overnight. The reaction mixture was
concentrated under a reduced pressure. The residue was dissolved in
300 ml ethyl acetate, and washed with 100 ml water. The organic
phase was dried with anhydrous sodium sulfate, filtered, and
evaporated to dryness under a reduced pressure to produce 18 g of a
product with a yield of 83%.
Step 3: Synthesis of tert-butyl
{5-amino-6-{[2-(dimethylamino)ethyl](methyl)amino}-2-isopropyloxypyridin--
3-yl}carbamate
##STR00040##
The compound was synthesized in the same manner as those in Step 2
of Intermediate 1a with a yield of 97%.
MS m/z: 368 [M+1].
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 7.61 (s, 1H), 7.44 (s, 1H),
6.74 (br s, 2H), 5.09-4.96 (m, 1H), 3.29 (t, J=5.8 Hz, 2H), 3.19
(t, J=5.7 Hz, 2H), 2.70 (s, 6H), 2.56 (s, 3H), 1.45 (s, 9H), 1.26
(d, J=6.2 Hz, 6H).
Step 4: Synthesis of tert-butyl
{5-acrylamide-6-{[2-(dimethylamino)ethyl](methyl)amino}-2-isopropyloxypyr-
idin-3-yl}carbamate
##STR00041##
To a 500 ml three-necked flask were added tert-butyl
{5-amino-6-{[2-(dimethylamino)ethyl](methyl)amino}-2-isopropyloxypyridin--
3-yl}carbamate (9 g, 24.49 mmol), trimethylamine (6.83 ml, 49.0
mmol) and 250 ml dichloromethane. The reaction mixture was cooled
in an ice-water bath to below 5.degree. C. Acryloyl chloride (2.1
ml, 25.7 mmol) was dropwisely added. The resulting mixture was
continued to react for 1 hour. The reaction mixture was washed
successively with 150 ml saturated sodium bicarbonate solution and
150 ml saturated brine, dried with anhydrous sodium sulfate, and
filtered. The filtrate was evaporated to dryness under a reduced
pressure to produce 5 g of a product with a yield of 48%. MS m/z:
422 [M+1].
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.76 (s, 1H), 8.16 (s, 1H),
7.88 (s, 1H), 6.44 (dd, J=17.0, 10.1 Hz, 1H), 6.22 (dd, J=17.0, 1.9
Hz, 1H), 5.74 (dd, J=10.1, 1.9 Hz, 1H), 5.22-5.13 (m, 1H), 3.09 (t,
J=6.5 Hz, 2H), 2.77 (s, 3H), 2.41 (t, J=6.5 Hz, 2H), 2.18 (s, 6H),
1.45 (s, 9H), 1.31 (d, J=6.2 Hz, 6H).
Intermediate 1e: tert-butyl
{5-acrylamide-6-{[2-(dimethylamino)ethyl](methyl)amino}-2-(2,2,2-trifluor-
oethoxyl)pyridin-3-yl}carbamate
##STR00042##
The compound was synthesized in the same manner as those in Step 1
of Intermediate 1d with a yield of 99%. MS m/z: 480 [M+1].
Step 2: Synthesis of tert-butyl
{6-{[2-(dimethylamino)ethyl](methyl)amino}-2-(2,2,2-trifluoroethoxyl)-5-n-
itropyridin-3-yl}carbamate
##STR00043##
The compound was synthesized in the same manner as those in Step 2
of Intermediate 1d with a yield of 88%. MS m/z: 438 [M+1].
Step 3: Synthesis of tert-butyl
{5-amino-6-{[2-(dimethylamino)ethyl](methyl)amino}-2-(2,2,2-trifluoroetho-
xyl)pyridin-3-yl}carbamate
##STR00044##
The compound was synthesized in the same manner as those in Step 2
of Intermediate 1a with a yield of 76%. MS m/z: 408 [M+1].
Step 4: Synthesis of tert-butyl
{5-acrylamide-6-{[2-(dimethylamino)ethyl](methyl)amino}-2-(2,2,2-trifluor-
oethoxyl)pyridin-3-yl}carbamate
##STR00045##
The compound was synthesized in the same manner as those in Step 4
of Intermediate 1d with a yield of 62%. MS m/z: 462 [M+1].
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 10.11 (s, 1H), 9.35 (s,
1H), 6.61 (s, 1H), 6.46 (dd, J=16.9, 1.7 Hz, 1H), 6.39-6.25 (m,
1H), 5.70 (dd, J=10.0, 1.8 Hz, 1H), 4.76 (q, J=8.5 Hz, 2H), 2.96
(s, 2H), 2.71 (s, 3H), 2.42 (s, 2H), 2.34 (s, 6H), 1.53 (s,
9H).
Intermediate 2a: 3-(2-chloropyrimidin-4-yl)-1-methyl-1H-indole
##STR00046##
To a 500 mL single-necked flask were added 2,4-dichloropyrimidine
(14.9 g, 100 mmol), 1-methyl-1H-indole (13 g, 100 mmol), 200 ml
1,2-dichloroethane and aluminium chloride (13.9 g, 120 mmol). The
mixture was stirred at 80.degree. C. for 1.5 hours. The reaction
mixture was cooled to room temperature in an ice bath. 120 ml
methanol and 400 ml water were added to quench the reaction. A
solid precipitated and was filtered. The filter cake was washed
with methanol, and dried in vacuum to produce 17.2 g of a product
with a yield of 71%. MS m/z: 244 [M+1], 246.
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.53 (d, J=5.5 Hz, 1H), 8.49
(s, 1H), 8.42 (dd, J=7.0, 1.5 Hz, 1H), 7.81 (d, J=5.5 Hz, 1H), 7.56
(dd, J=7.0, 1.2 Hz, 1H), 7.33-7.26 (m, 2H), 3.90 (d, J=5.2 Hz,
3H).
Intermediate 2b:
3-(2,5-dichloropyrimidin-4-yl)-1-methyl-1H-indole
##STR00047##
The compound was synthesized in the same manner as those in
Intermediate 2a with a yield of 87%. MS m/z: 278[M+1], 279,
280.
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.79 (s, 1H), 8.74 (s, 1H),
8.56 (dd, J=7.3, 1.2 Hz, 1H), 7.62 (d, J=7.6 Hz, 1H), 7.39-7.34 (m,
1H), 7.34-7.29 (m, 1H), 3.97 (s, 3H).
Intermediate 2c:
3-(2-chloropyrimidin-4-yl)-1-methyl-5-fluoro-1H-indole
##STR00048##
The compound was synthesized in the same manner as those in
Intermediate 2a with a yield of 29%. MS m/z: 262 [M+1], 264.
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.55 (s, 1H), 8.53 (d, J=5.5
Hz, 1H), 8.10 (dd, J=10.3, 2.5 Hz, 1H), 7.80 (d, J=5.5 Hz, 1H),
7.60 (dd, J=8.9, 4.6 Hz, 1H), 7.17 (td, J=9.1, 2.6 Hz, 1H), 3.90
(s, 3H).
Intermediate 2d:
3-(2-chloropyrimidin-4-yl)-1-methyl-6-fluoro-1H-indole
##STR00049##
The compound was synthesized in the same manner as those in
Intermediate 2a. MS m/z: 262 [M+1], 264.
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.54 (d, J=5.5 Hz, 1H), 8.49
(s, 1H), 8.39 (dd, J=8.8, 5.6 Hz, 1H), 7.81 (d, J=5.5 Hz, 1H), 7.47
(dd, J=9.9, 2.3 Hz, 1H), 7.14 (td, J=9.6, 2.4 Hz, 1H), 3.86 (s,
3H).
Intermediate 2e:
3-(2-chloropyrimidin-4-yl)-1-methyl-5,6-difluoro-1H-indole
##STR00050##
The compound was synthesized in the same manner as those in
Intermediate 2a. MS m/z: 280 [M+1], 282.
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.54 (d, J=5.5 Hz, 1H), 8.52
(s, 1H), 8.22 (dd, J=11.7, 8.2 Hz, 1H), 7.79 (d, J=5.5 Hz, 1H),
7.73 (dd, J=11.2, 7.0 Hz, 1H), 3.86 (s, 3H).
Intermediate 2f:
3-(2,5-dichloropyrimidin-4-yl)-1-methyl-6-fluoro-1H-indole
##STR00051##
The compound was synthesized in the same manner as those in
Intermediate 2a. MS m/z: 296 [M+1], 297, 298.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.69 (dd, J=8.9, 5.5 Hz,
1H), 8.50 (s, 1H), 8.41 (s, 1H), 7.17 7.07 (m, 2H), 3.90 (s,
3H).
Intermediate 2g:
3-(2,5-dichloropyrimidin-4-yl)-1-methyl-5,6-difluoro-1H-indole
##STR00052##
The compound was synthesized in the same manner as those in
Intermediate 2a. MS m/z: 314 [M+1], 315, 316.
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.85 (s, 1H), 8.77 (s, 1H),
8.39 (dd, J=12.1, 8.3 Hz, 1H), 7.83 (dd, J=11.0, 7.1 Hz, 1H), 3.94
(s, 3H).
Intermediate 2h:
3-(2,5-dichloropyrimidin-4-yl)-1-methyl-5-fluoro-1H-indole
##STR00053##
The compound was synthesized in the same manner as those in
Intermediate 2a. MS m/z: 296 [M+1], 297, 298.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.49 (s, 1H), 8.46 (s,
1H), 8.46-8.42 (m, 1H), 7.34 (dd, J=8.9, 4.4 Hz, 1H), 7.14 (td,
J=8.9, 2.6 Hz, 1H), 3.94 (s, 3H).
Intermediate 2i:
3-(2-chloro-5-fluoropyrimidin-4-yl)-1-methyl-1H-indole
##STR00054##
The compound was synthesized in the same manner as those in
Intermediate 2a with a yield of 73%. MS m/z: 262 [M+1], 264.
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.69 (d, J=3.7 Hz, 1H), 8.54
(dd, J=7.2, 1.2 Hz, 1H), 8.39 (d, J=3.0 Hz, 1H), 7.62 (d, J=7.5 Hz,
1H), 7.41-7.30 (m, 2H), 3.96 (s, 3H).
Intermediate 2j:
3-(2-chloro-5-fluoropyrimidin-4-yl)-1-methyl-5-fluoro-1H-indole
##STR00055##
The compound was synthesized in the same manner as those in
Intermediate 2a with a yield of 77%. MS m/z: 280 [M+1], 282.
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.71 (d, J=3.5 Hz, 1H), 8.45
(d, J=2.8 Hz, 1H), 8.20 (dd, J=10.3, 2.5 Hz, 1H), 7.66 (dd, J=8.9,
4.5 Hz, 1H), 7.30-7.16 (m, 1H), 3.96 (s, 3H).
Intermediate 2k:
3-(2-chloro-5-fluoropyrimidin-4-yl)-1-methyl-5,6-difluoro-1H-indole
##STR00056##
The compound was synthesized in the same manner as those in
Intermediate 2a. MS m/z: 298 [M+1], 300.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.56 (dd, J=11.4, 8.1 Hz,
1H), 8.36 (d, J=3.3 Hz, 1H), 8.01 (d, J=2.6 Hz, 1H), 7.19 (dd,
J=10.1, 6.6 Hz, 1H), 3.90 (s, 3H).
Intermediate 2:
.[.3-(2,5-dichloropyrimidin-4-yl)-1-methyl-1H-pyrro[2,3-b]pyridine.].
.Iadd.3-(2,5-dichloropyrimidin-4-yl)-1-methyl-1H-pyrrolo[2,3-b]pyridine.I-
addend.
##STR00057##
Step 1: Synthesis of
.[.3-bromo-1-p-tosyl-1H-pyrro[2,3-b]pyridine.].
.Iadd.3-bromo-1-p-tosyl-1H-pyrrolo[2,3-b]pyridine.Iaddend.
##STR00058##
To a 250 mL three-necked flask were added
.[.3-bromo-1H-pyrro[2,3-b]pyridine.].
.Iadd.3-bromo-1H-pyrrolo[2,3-b]pyridine .Iaddend.(4.0 g, 20.3 mmol)
and 80 ml tetrahydrofuran. The mixture was cooled to below
5.degree. C. in an ice-water bath. 60% of sodium hydride (1.3 g,
32.5 mmol) was added. The mixture was stirred for 15 minutes.
p-Toluensulfonyl chloride (4.1 g, 21.3 mmol) was added. The
reaction continued for 15 minutes. 150 ml water was added to quench
the reaction. The reaction mixture was extracted with ethyl acetate
(150 ml). The organic layer was evaporated to dryness under a
reduced pressure to produce a brown solid, which was added to
petroleum ether to form a slurry, and a brown solid (5 g) was
obtained with a yield of 70%. MS m/z: 351 [M+1], 353.
Step 2: Synthesis of
.[.3-(2,5-dichloropyrimidin-4-yl)-1-p-tosyl-1H-pyrro[2,3-b]pyridine.].
.Iadd.3-(2,5-dichloropyrimidin-4-yl)-1-p-tosyl-1H-pyrrolo[2,3-b]pyridine.-
Iaddend.
##STR00059##
To a 100 mL single-necked flask were added
.[.3-bromo-1-p-tosyl-1H-pyrro[2,3-b]pyridine.].
.Iadd.3-bromo-1-p-tosyl-1H-pyrrolo[2,3-b]pyridine .Iaddend.(2.0 g,
5.7 mmol), bis(pinacolato)diboron (1.9 g, 7.4 mmol), potassium
acetate (1.7 g, 17.1 mmol),
[1,1'-Bis(diphenylphosphino)ferrocene]dichloropalladium (0.21 g,
0.285 mmol) and 25 ml dioxane with atmosphere replaced by argon.
The mixture was reacted at 85.degree. C. for 6.5 hours. LC-MS
monitoring showed the starting materials were depleted. To the
reaction mixture was added 2,4,5-trichloropyrimidine (1.3 g, 7.0
mmol), 5 ml 2N sodium carbonate solution and
[1,1'-Bis(diphenylphosphino)ferrocene]dichloropalladium (0.37 g,
0.50 mmol) with atmosphere replaced by argon. The reaction
continued at 85.degree. C. overnight. The reaction mixture was
diluted with 150 ml ethyl acetate, and washed with 150 ml water.
The aqueous phase was extracted with dichloromethane (120
ml.times.3). The organic phases were combined, dried with anhydrous
sodium sulfate, and filtered. The filtrate was evaporated to
dryness under a reduced pressure, and purified by silica gel column
chromatography (petroleum ether:ethyl acetate=5:1). The product was
added to a mixed solvent of petroleum ether and ethyl acetate
(volume ratio=2:1) to form a slurry, and 1.0 g of an off-white
solid was obtained with a yield of 42%. MS m/z: 419 [M+1], 421.
Step 3: Synthesis of
.[.3-(2,5-dichloropyrimidin-4-yl)-1H-pyrro[2,3-b]pyridine.].
.Iadd.3-(2,5-dichloropyrimidin-4-yl)-1H-pyrrolo[2,3-b]pyridine.Iaddend.
##STR00060##
To a 100 mL single-necked flask were added
.[.3-(2,5-dichloropyrimidin-4-yl)-1-p-tosyl-1H-pyrro[2,3-b]pyridine.].
.Iadd.3-(2,5-dichloropyrimidin-4-yl)-1-p-tosyl-1H-pyrrolo[2,3-b]pyridine
.Iaddend.(0.95 g, 2.3 mmol) and 30 ml tetrahydrofuran. Under
stirring, tetrabutylammonium fluoride (1.2 g, 4.6 mmol) was added.
The mixture was reacted at room temperature for 20 minutes. To the
reaction mixture was added 100 ml ethyl acetate. The reaction
mixture was washed with 100 ml water. The organic phase was dried
with anhydrous sodium sulfate, and filtered. The filtrate was
evaporated to dryness under a reduced pressure. The residue was
added to 20 ml of a mixed solvent of petroleum ether and ethyl
acetate (volume ratio=4:1) to form a slurry. The slurry was
filtered by suction to produce 500 mg of an off-white solid with a
yield of 83%. MS m/z: 265 [M+1].
Step 4: Synthesis of
.[.3-(2,5-dichloropyrimidin-4-yl)-1-methyl-1H-pyrro[2,3-b]pyridine.].
.Iadd.3-(2,5-dichloropyrimidin-4-yl)-1-methyl-1H-pyrrolo[2,3-b]pyridine.I-
addend.
##STR00061##
To a 50 ml three-necked flask were added
.[.3-(2,5-dichloropyrimidin-4-yl)-1H-pyrro[2,3-b]pyridine.].
.Iadd.3-(2,5-dichloropyrimidin-4-yl)-1H-pyrrolo[2,3-b]pyridine
.Iaddend.(480 mg, 1.8 mmol) and 15 ml N,N-dimethylformamide. The
resulting mixture was cooled to 5.degree. C. under an ice-water
bath. 60% of sodium hydride (145 mg, 3.6 mmol) was added. The
mixture was stirred for 10 minutes, and methyl iodide (0.12 ml, 1.9
mmol) was added thereto. The resulting mixture was stirred at
5.degree. C. for 15 minutes. The reaction mixture was poured to
ice-water, and a solid precipitated and was filtered by suction.
The filter cake was dried to produce 450 mg of .[.an.]. .Iadd.a
.Iaddend.beige solid with a yield of 89%. MS m/z: 265 [M+1].
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.94 (s, 1H), 8.81 (dd,
J=8.0, 1.6 Hz, 1H), 8.78 (s, 1H), 8.44 (dd, J=4.7, 1.6 Hz, 1H),
7.38 (dd, J=8.0, 4.7 Hz, 1H), 3.97 (s, 3H).
Intermediate 2m:
.[.5-(2,5-dichloropyrimidin-4-yl)-1-methyl-1H-pyrro[2,3-b]pyridine.].
.Iadd.5-(2,5-dichloropyrimidin-4-yl)-1-methyl-1H-pyrrolo[2,3-b]pyridine.I-
addend.
##STR00062##
The compound was synthesized in the same manner as those in Step 2
of Intermediate 2l with a yield of 50%. MS m/z: 279 [M+1].
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.99 (s, 1H), 8.75 (d, J=2.1
Hz, 1H), 8.51 (d, J=2.1 Hz, 1H), 7.68 (d, J=3.5 Hz, 1H), 6.66 (d,
J=3.5 Hz, 1H), 3.90 (s, 3H).
Intermediate 2n:
2,5-dichloro-4-(1-methyl-1H-pyrazol-4-yl)pyrimidine
##STR00063##
To a three-necked flask were added 2,4,5-trichloropyrimidine (2.0
g, 10.9 mmol), 1-methyl-4-pyrazole-bis(pinacolato)diboron (1.75 g,
8.4 mmol), 8.4 ml 2N sodium carbonate solution,
[1,1'-Bis(diphenylphosphino)ferrocene]dichloropalladium (0.61 g,
0.84 mmol) and 30 ml dioxane with atmosphere replaced by argon. The
mixture was stirred at 80.degree. C. overnight. To the reaction
mixture was added 150 ml ethyl acetate, washed successively with
150 ml water and 100 ml saturated sodium chloride solution, dried
with anhydrous sodium sulfate, and evaporated to dryness under a
reduced pressure to produce a earth yellow solid (1.6 g) with a
yield of 83%. MS m/z: 229 [M+1].
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.82 (s, 1H), 8.75 (s, 1H),
8.27 (s, 1H), 3.96 (s, 3H).
Intermediate 2o: 2,5-dichloro-2'-methoxy-4,5'-bipyrimidine
##STR00064##
The compound was synthesized in the same manner as those in
Intermediate 2n with a yield of 70%. MS m/z: 257 [M+1].
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.10 (s, 2H), 9.05 (s, 1H),
4.04 (s, 3H).
Intermediate 2p: 2,5-dichloro-2'-amino-4,5'-bipyrimidine
##STR00065##
The compound was synthesized in the same manner as those in
Intermediate 2n with a yield of 44%. MS m/z: 242 [M+1].
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 8.90 (s, 1H), 8.84 (s, 2H),
7.52 (s, 2H).
Example 1:
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{-
5-chloro-[4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acry-
lamide
##STR00066##
Step 1: Synthesis of
N.sup.2-methyl-N.sup.2-[2-(dimethylamino)ethyl]-6-isopropyloxy-N.sup.5
[5-chloro-4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]-3-nitropyridin-2,5-di-
amine
##STR00067##
To a 50 mL single-necked flask were added
N.sup.2-methyl-N.sup.2-[2-(dimethylamino)ethyl]-6-isopropyloxy-3-nitropyr-
idin-2,5-diamine (490 mg, 1.65 mmol),
3-(2,5-dichloropyrimidin-4-yl)-1-methyl-1H-indole (550 mg, 1.98
mmol), tris(dibenzylideneacetone)dipalladium (226 mg, 0.2475 mmol),
4,5-Bis(diphenylphosphino)-9,9-dimethylxanthene (286 mg, 0.495
mmol), potassium phosphate (874 mg, 4.125 mmol) and 15 ml dioxane.
Under the nitrogen protection, the mixture was reacted at
100.degree. C. overnight. The reaction mixture was filtered with
diatomite. The filtrate was evaporated to dryness under a reduced
pressure, purified by silica gel column chromatography
(dichloromethane:methanol=50:1) to produce 480 mg of a product with
a yield of 54%. MS m/z: 539 [M+1].
Step 2: Synthesis of
N.sup.2-methyl-N.sup.2-[2-(dimethylamino)ethyl]-6-isopropyloxy-N.sup.5-[5-
-chloro-4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]pyridin-2,3,5-triamine
##STR00068##
To a 50 mL single-necked flask were added
N.sup.2-methyl-N.sup.2-[2-(dimethylamino)ethyl]-6-isopropyloxy-N.sup.5-[5-
-chloro-4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]-3-nitropyridin-2,5-diami-
ne (480 mg, 0.892 mmol), ammonia chloride (48 mg, 0.897 mmol) and
12 ml of a mixed solvent of ethanol and water (volume ratio=3:1).
To the mixture was added in batch a reduced iron powders (240 mg,
4.26 mmol). The mixture was stirred at 80.degree. C. for 1 hour.
The reaction mixture was cooled to room temperature, and filtered
through diatomite. The filtrate was evaporated to dryness under a
reduced pressure, dissolved in dichloromethane, and washed with a
saturated sodium carbonate solution. The organic layer was dried
with anhydrous sodium sulfate and filtered. The filtrate was
evaporated to dryness under a reduced pressure, and subjected to a
preparative TLC separation (dichloromethane:ethyl
acetate:methanol=5:5:1) to produce 96 mg of a product with a yield
of 43%. MS m/z: 509 [M+1].
Step 3: Synthesis of
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{5-chloro-[-
4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylamide
##STR00069##
To a 50 ml single-necked flask were added
N.sup.2-methyl-N.sup.2-[2-(dimethylamino)ethyl]-6-isopropyloxy-N.sup.5-[5-
-chloro-4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]pyridin-2,3,5-triamine
(196 mg, 0.385 mmol) and 10 ml dichloromethane. The reaction
mixture was cooled in an ice-water bath. 0.5 N of a solution of
acryloyl chloride in dichloromethane (0.8 ml, 0.4 mmol) and
triethylamine (0.15 ml, 1.08 mmol) were added. The mixture was
reacted at room temperature for 0.5 hour. To the reaction mixture
was added a suitable amount of water. The dichloromethane layer was
separated, dried with anhydrous sodium sulfate, and filtered. The
filtrate was concentrated under a reduced pressure, and purified by
preparative TLC separation (dichloromethane:ethyl
acetate:methanol=5:5:1) to produce 130 mg of a pale-yellow solid
with a yield of 60%. MS m/z: 563 [M+1], 565.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.75 (s, 1H), 9.36 (s,
1H), 8.39 (s, 1H), 8.38-8.33 (m, 1H), 8.29 (s, 1H), 7.40 (s, 1H),
7.38-7.33 (m, 1H), 7.33-7.27 (m, 2H), 7.06 (dd, J=16.9, 10.2 Hz,
1H), 6.39 (d, J=16.9 Hz, 1H), 5.70 (d, J=10.2 Hz, 1H), 5.29-5.20
(m, 1H), 3.90 (s, 3H), 3.51-3.46 (m, 2H), 3.09 (br s, 2H), 2.77 (s,
3H), 2.75 (s, 6H), 1.38 (d, J=6.2 Hz, 6H).
Example 2:
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{-
[4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylamide
##STR00070##
Step 1: Synthesis of
N.sup.2-methyl-N.sup.2-[2-(dimethylamino)ethyl]-6-isopropyloxy-N.sup.5-[4-
-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]-3-nitropyridin-2,5-diamine
##STR00071##
The compound was synthesized in the same manner as those in Step 1
of Example 1 with a yield of 100%.
Step 2: Synthesis of
N.sup.2-methyl-N.sup.2-[2-(dimethylamino)ethyl]-6-isopropyloxy-N.sup.5-[4-
-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]pyridin-2,3,5-triamine
##STR00072##
N.sup.2-methyl-N.sup.2-[2-(dimethylamino)ethyl]-6-isopropyloxy-N.sup.5-[4-
-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]-3-nitropyridin-2,5-diamine
(200 mg, 0.397 mmol) was dissolved in 12 ml methanol. 35 mg
platinum dioxide was added and hydrogen was introduced. The
resulting mixture was stirred at room temperature for 1.5 hour, and
filtered. The filtrate was concentrated under a reduced pressure,
and subjected to a preparative TLC seperation
(dichloromethane:ethyl acetate:methanol=9:1:1) to produce 50 mg of
a product with a yield of 27%. MS m/z: 475 [M+1].
Step 3: Synthesis of
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[4-(1-meth-
yl-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylamide
##STR00073##
The compound was synthesized in the same manner as those in Step 3
of Example 1 with a yield of 45%. MS m/z: 529 [M+1].
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.80 (s, 1H), 9.73 (s,
1H), 8.88 (s, 1H), 8.39 (d, J=5.3 Hz, 1H), 8.11-8.03 (m, 1H), 7.81
(d, J=8.3 Hz, 1H), 7.48 (s, 1H), 7.42-7.40 (m, 1H), 7.36 (d, J=8.1
Hz, 1H), 7.30 (d, J=3.7 Hz, 1H), 7.24 (d, J=5.3 Hz, 1H), 6.50 (dd,
J=16.9, 1.9 Hz, 1H), 5.76 (dd, J=10.2, 1.9 Hz, 1H), 5.32-5.21 (m,
1H), 3.99 (s, 3H), 3.52 (br s, 2H), 3.11 (br s, 2H), 2.81 (d, J=2.5
Hz, 9H), 1.39 (d, J=6.2 Hz, 6H).
Example 3:
.[.N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-(2,2,2-triflu-
oroethoxyl)-5-{[4-(1-methyl-H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl-
}acrylamide.].
.Iadd.N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-(2,2,2-trifluoroetho-
xyl)-5-{[4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acryl-
amide.Iaddend.
##STR00074##
Step 1: Synthesis of
N.sup.2-methyl-N.sup.2-[2-(dimethylamino)ethyl]-6-(2,2,2-trifluoroethoxyl-
)-N.sup.5-[4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]-3-nitropyridin-2,5-di-
amine
##STR00075##
The compound was synthesized in the same manner as those in Step 1
of Example 1 with a yield of 86%. MS m/z: 545 [M+1].
Step 2: Synthesis of
N.sup.2-methyl-N.sup.2-[2-(dimethylamino)ethyl]-6-(2,2,2-trifluoroethoxyl-
)-N.sup.5-[4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]pyridin-2,3,5-triamine
##STR00076##
The compound was synthesized in the same manner as those in Step 2
of Example 2 with a yield of 56%. MS m/z: 515 [M+1].
Step 3: Synthesis of
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-(2,2,2-trifluoroethoxyl)-5-
-{[4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylamide
##STR00077##
The compound was synthesized in the same manner as those in Step 3
of Example 1 with a yield of 23%. MS m/z: 569 [M+1].
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 10.41 (s, 1H), 10.27 (s,
1H), 8.68 (s, 1H), 8.44 (s, 1H), 8.28 (t, J=8.5 Hz, 2H), 8.18 (s,
1H), 7.52 (d, J=8.0 Hz, 1H), 7.29-7.14 (m, 3H), 6.98 (s, 1H), 6.28
(d, J=17.1 Hz, 1H), 5.76 (d, J=10.4 Hz, 1H), 5.00 (q, J=9.0 Hz,
2H), 3.89 (s, 3H), 3.61 (s, 2H), 3.28 (s, 2H), 2.80 (s, 3H), 2.73
(s, 6H).
Example 4:
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-(2,2,2-trifluoro-
ethoxyl)-5-{5-chloro-[4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]amino}pyrid-
in-3-yl}acrylamide
##STR00078##
Step 1: Synthesis of
N.sup.2-methyl-N.sup.2-[2-(dimethylamino)ethyl]-6-(2,2,2-trifluoroethoxyl-
)-N.sup.5-[5-chloro-4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]-3-nitropyrid-
in-2,5-diamine
##STR00079##
The compound was synthesized in the same manner as those in Step 1
of Example 1 with a yield of 86%.
Step 2: Synthesis of
N.sup.2-methyl-N.sup.2-[2-(dimethylamino)ethyl]-6-(2,2,2-trifluoroethoxyl-
)-N.sup.5-[5-chloro-4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]pyridin-2,3,5-
-triamine
##STR00080##
The compound was synthesized in the same manner as those in Step 2
of Example 1 with a yield of 65%.
Step 3: Synthesis of
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-(2,2,2-trifluoroethoxyl)-5-
-{5-chloro-[4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}ac-
rylamide
##STR00081##
The compound was synthesized in the same manner as those in Step 3
of Example 1 with a yield of 15%. MS m/z: 603 [M+1], 605.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 11.68 (br s, 1H), 9.77
(s, 1H), 9.48 (s, 1H), 8.42 (s, 1H), 8.38 (d, J=8.7 Hz, 1H), 8.33
(s, 1H), 7.40-7.37 (m, 2H), 7.32 (dd, J=6.7, 3.0 Hz, 2H), 7.12 (dd,
J=16.8, 10.2 Hz, 1H), 6.43 (dd, J=16.9, 1.8 Hz, 1H), 5.72 (dd,
J=10.2, 1.8 Hz, 1H), 4.83 (q, J=8.4 Hz, 2H), 3.93 (s, 3H), 3.60 (s,
2H), 3.17 (s, 2H), 2.86 (s, 3H), 2.85 (s, 6H).
Example 5:
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{-
[4-(1-methyl-5-fluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acry-
lamide
##STR00082##
Step 1: Synthesis of
N.sup.2-methyl-N.sup.2-[2-(dimethylamino)ethyl]-6-isopropyloxy-N.sup.5-[4-
-(1-methyl-5-fluoro-1H-indol-3-yl)pyrimidin-2-yl]-3-nitropyridin-2,5-diami-
ne
##STR00083##
The compound was synthesized in the same manner as those in Step 1
of Example 1 with a yield of 57%. MS m/z: 523.
Step 2: Synthesis of
N.sup.2-methyl-N.sup.2-[2-(dimethylamino)ethyl]-6-isopropyloxy-N.sup.5-[4-
-(1-methyl-5-fluoro-1H-indol-3-yl)pyrimidin-2-yl]pyridin-2,3,5-triamine
##STR00084##
The compound was synthesized in the same manner as those in Step 2
of Example 1 with a yield of 64%. MS m/z: 493 [M+1].
Step 3: Synthesis of
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[4-(1-meth-
yl-5-fluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylamide
##STR00085##
The compound was synthesized in the same manner as those in Step 3
of Example 1 with a yield of 45%. MS m/z: 547 [M+1].
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.82 (s, 1H), 9.80 (s,
1H), 8.93 (s, 1H), 8.40 (d, J=5.2 Hz, 1H), 7.71 (d, J=9.7 Hz, 1H),
7.49 (s, 1H), 7.32 (dd, J=8.8, 4.4 Hz, 1H), 7.20-6.98 (m, 3H), 6.48
(d, J=16.8 Hz, 1H), 5.76 (d, J=10.5 Hz, 1H), 5.31-5.25 (m, 1H),
3.99 (s, 3H), 3.43 (br s, 2H), 2.98 (br s, 2H), 2.71 (s, 6H), 1.39
(d, J=6.1 Hz, 6H).
Example 6:
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{-
[4-(1-methyl-5,6-difluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}-
acrylamide
##STR00086##
Step 1: Synthesis of
N.sup.2-methyl-N.sup.2-[2-(dimethylamino)ethyl]-6-isopropyloxy-N.sup.5-[4-
-(1-methyl-5,6-difluoro-1H-indol-3-yl)pyrimidin-2-yl]-3-nitropyridin-2,5-d-
iamine
##STR00087##
The compound was synthesized in the same manner as those in Step 1
of Example 1 with a yield of 28%. MS m/z: 541.
Step 2: Synthesis of
N.sup.2-methyl-N.sup.2-[2-(dimethylamino)ethyl]-6-isopropyloxy-N.sup.5-[4-
-(1-methyl-5,6-difluoro-1H-indol-3-yl)pyrimidin-2-yl]pyridin-2,3,5-triamin-
e
##STR00088##
The compound was synthesized in the same manner as those in Step 2
of Example 1 with a yield of 64%. MS m/z: 511 [M+1].
Step 3: Synthesis of
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[4-(1-meth-
yl-5,6-difluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylamide
##STR00089##
The compound was synthesized in the same manner as those in Step 3
of Example 1 with a yield of 38%. MS m/z: 565 [M+1].
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.73 (s, 1H), 9.70 (s,
1H), 8.82 (s, 1H), 8.39 (d, J=4.9 Hz, 1H), 7.88-7.74 (m, 1H), 7.50
(s, 1H), 7.25 (dd, J=16.2, 9.7 Hz, 1H), 7.20-7.13 (m, 1H), 7.05 (d,
J=5.0 Hz, 1H), 6.47 (d, J=16.5 Hz, 1H), 5.76 (d, J=10.3 Hz, 1H),
5.31-5.21 (m, 1H), 3.94 (s, 3H), 3.54 (s, 2H), 3.13 (s, 2H), 2.82
(s, 6H), 1.39 (d, J=5.9 Hz, 6H).
Example 7:
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{-
5-chloro-[4-(1-methyl-6-fluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin--
3-yl}acrylamide
##STR00090##
To a 25 ml three-necked flask were added tert-butyl
{5-acrylamide-6-{[2-(dimethylamino)ethyl](methyl)amino}-2-isopropyloxypyr-
idin-3-yl}carbamate (160 mg, 0.38 mmol),
3-(2,5-dichloropyrimidin-4-yl)-6-fluoro-1-methyl-1H-indole (112 mg,
0.38 mmol), p-toluenesulfonic acid monohydrate (112 mg, 0.59 mmol),
4 ml 2-amyl alcohol and 2 ml N-methylpyrrolidone. Under the
nitrogen protection, the mixture was reacted at 120.degree. C.
overnight. The mixture was cooled to room temperature and poured
into 50 ml water. A solid precipitated and was filtered. The solid
was dissolved in 20 ml dichloromethane, washed successively with 10
ml saturated sodium bicarbonate solution and 10 ml water, dried
with anhydrous sodium sulfate, and filtered. The filtrate was
evaporated to dryness under a reduced pressure, and subjected to a
preparative TLC seperation (dichloromethane:methanol=10:1).
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.78 (s, 1H), 9.46 (s,
1H), 8.43 (s, 1H), 8.32-8.28 (m, 2H), 7.40 (s, 1H), 7.08-7.03 (m,
2H), 7.00-6.86 (m, 1H), 6.45-6.38 (m, 1H), 5.73 (d, J=10.2 Hz, 1H),
5.31-5.23 (m, 1H), 3.88 (s, 3H), 3.45 (s, 2H), 2.99 (s, 2H), 2.80
(s, 3H), 2.73 (s, 6H), 1.39 (d, J=6.2 Hz, 6H).
Example 8:
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{-
5-chloro-[4-(1-methyl-5,6-difluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyri-
din-3-yl}acrylamide
##STR00091##
The compound was prepared in the same manner as those in Example 7
with a yield of 8%. MS m/z: 599 [M+1].
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.93 (s, 1H), 8.71 (s, 1H),
8.66 (s, 1H), 8.37 (s, 1H), 8.25 8.15 (m, 1H), 8.11 (s, 1H), 7.68
(dd, J=11.1, 7.0 Hz, 1H), 6.83-6.64 (m, 1H), 6.21 (d, J=16.5 Hz,
1H), 5.73 (d, J=11.9 Hz, 1H), 5.21 5.13 (m, 1H), 3.89 (s, 3H), 3.36
(s, 4H), 2.80 (s, 3H), 2.56 (s, 6H), 1.18 (d, J=6.1 Hz, 6H).
Example 9:
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{-
5-chloro-[4-(1-methyl-5-fluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin--
3-yl}acrylamide
##STR00092##
The compound was prepared in the same manner as those in Example 7
with a yield of 10%. MS m/z: 581 [M+1].
.sup.1H NMR (400 MHz, MeOD) .delta. 8.48 (s, 1H), 8.36 (s, 1H),
8.34 (s, 1H), 8.02 (dd, J=10.6, 2.4 Hz, 1H), 7.44 (dd, J=8.9, 4.4
Hz, 1H), 7.03 (td, J=9.0, 2.5 Hz, 1H), 6.47 (dd, J=17.0, 9.3 Hz,
1H), 6.40 (dd, J=17.0, 2.5 Hz, 1H), 5.82 (dd, J=9.3, 2.5 Hz, 1H),
5.39-5.28 (m, 1H), 3.91 (s, 3H), 3.74 (t, J=5.7 Hz, 2H), 3.32 (t,
J=5.9 Hz, 2H), 2.89 (s, 6H), 2.80 (s, 3H), 1.37 (d, J=6.2 Hz,
6H).
Example 10:
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{5-fluoro-[-
4-(1-methyl-5-fluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acryl-
amide
##STR00093##
The compound was prepared in the same manner as those in Example 7
with a yield of 9%. MS m/z: 565 [M+1].
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 10.01 (s, 1H), 8.45 (s, 1H),
8.35 (d, J=3.9 Hz, 1H), 8.28 (d, J=2.7 Hz, 1H), 8.26 (s, 1H), 8.07
(d, J=10.2 Hz, 1H), 7.55 (dd, J=8.9, 4.6 Hz, 1H), 7.11 (td, J=9.1,
2.6 Hz, 1H), 6.90 (s, 1H), 6.23 (dd, J=17.1, 1.9 Hz, 1H), 5.72 (dd,
J=10.2, 1.9 Hz, 1H), 5.26-5.12 (m, 1H), 3.92 (s, 3H), 3.53 (s, 2H),
3.24 (s, 2H), 2.77 (s, 3H), 2.71 (s, 6H), 1.21 (d, J=6.2 Hz,
6H).
Example 11:
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{5-fluoro-[-
4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylamide
##STR00094##
The compound was prepared in the same manner as those in Example 7
with a yield of 7%. MS m/z: 547 [M+1].
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 10.12 (s, 1H), 8.47 8.32 (m,
3H), 8.23 (d, J=2.8 Hz, 1H), 8.19 (s, 1H), 7.52 (d, J=8.2 Hz, 1H),
7.24 (t, J=7.6 Hz, 1H), 7.15 (t, J=7.5 Hz, 1H), 6.88 (dd, J=16.9,
10.3 Hz, 1H), 6.23 (dd, J=17.1, 1.8 Hz, 1H), 5.72 (dd, J=10.2, 1.7
Hz, 1H), 5.27-5.15 (m, 1H), 4.04 (s, 3H), 3.91 (s, 3H), 3.57 (s,
2H), 3.28 (s, 2H), 2.76 (s, 6H), 1.26 (d, J=6.2 Hz, 6H).
Example 12:
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{5-fluoro-[-
4-(1-methyl-5,6-difluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}a-
crylamide
##STR00095##
The compound was prepared in the same manner as those in Example 7.
MS m/z: 583 [M+1].
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.70 (s, 1H), 9.47 (s,
1H), 8.29 (d, J=3.4 Hz, 2H), 8.23 (dd, J=11.8, 8.2 Hz, 1H), 7.33
(s, 1H), 7.14 (dd, J=10.3, 6.7 Hz, 2H), 6.41 (dd, J=16.9, 1.8 Hz,
1H), 5.73 (dd, J=10.2, 1.8 Hz, 1H), 5.31-5.23 (m, 1H), 3.90 (s,
3H), 3.55 (s, 2H), 3.13 (s, 2H), 2.83 (s, 9H), 1.40 (d, J=6.2 Hz,
6H).
Example 13:
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[4-(1-meth-
yl-6-fluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylamide
##STR00096##
The compound was prepared in the same manner as those in Example 7
with a yield of 15%. MS m/z: 547 [M+1].
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.77 (s, 1H), 9.76 (s,
1H), 8.82 (s, 1H), 8.39 (d, J=5.3 Hz, 1H), 7.98 (dd, J=8.7, 5.2 Hz,
1H), 7.47 (s, 1H), 7.16 (d, J=5.3 Hz, 1H), 7.08 (dd, J=9.6, 2.3 Hz,
1H), 7.03 (dd, J=9.1, 2.2 Hz, 1H), 6.49 (dd, J=16.9, 2.0 Hz, 1H),
5.77 (dd, J=10.2, 2.0 Hz, 1H), 5.27 (hept, J=6.2 Hz, 1H), 3.94 (s,
3H), 3.52 (s, 2H), 3.10 (s, 2H), 2.82 (s, 3H), 2.80 (s, 6H), 1.39
(d, J=6.2 Hz, 6H).
Example 14:
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-(2,2,2-trifluoroethoxyl)-5-
-{5-fluoro-[4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}ac-
rylamide
##STR00097##
The compound was prepared in the same manner as those in Example 7.
MS m/z: 587 [M+1].
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.87 (s, 1H), 9.53 (s,
1H), 8.52-8.44 (m, 1H), 8.28 (d, J=3.7 Hz, 1H), 8.21 (s, 1H), 7.38
(dd, J=8.1, 4.9 Hz, 1H), 7.33 (dd, J=6.0, 3.3 Hz, 2H), 7.19 (dd,
J=16.9, 10.3 Hz, 1H), 6.43 (dd, J=16.9, 1.5 Hz, 1H), 5.74 (dd,
J=10.3, 1.5 Hz, 1H), 4.83 (q, J=8.5 Hz, 2H), 3.93 (s, 3H), 3.56 (t,
J=5.1 Hz, 2H), 3.15 (t, J=5.1 Hz, 2H), 2.85 (s, 3H), 2.81 (s,
6H).
Example 15:
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-(2,2,2-trifluoroethoxyl)-5-
-{[4-(1-methyl-5-fluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}ac-
rylamide
##STR00098##
The compound was prepared in the same manner as those in Example 7.
MS m/z: 587 [M+1].
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.85 (s, 2H), 8.83 (s,
1H), 8.40 (d, J=5.3 Hz, 1H), 7.71 (dd, J=10.2, 2.1 Hz, 1H), 7.41
(s, 1H), 7.31 (dd, J=8.9, 4.5 Hz, 1H), 7.13 (d, J=5.3 Hz, 2H), 7.03
(td, J=9.0, 2.3 Hz, 1H), 6.49 (dd, J=16.9, 1.8 Hz, 1H), 5.78 (dd,
J=10.2, 1.8 Hz, 1H), 4.83 (q, J=8.5 Hz, 2H), 3.97 (s, 3H), 3.49 (s,
2H), 3.05 (s, 2H), 2.83 (s, 3H), 2.75 (s, 6H).
Example 16:
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{5-chloro-[-
4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylamide
methanesulfonate
##STR00099##
To
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{5-chlor-
o-[4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylamide
(56 mg, 0.1 mmol) were added 2 ml acetone, 0.4 ml water, and
methanesulfonic acid (6.5 .mu.l, 0.1 mmol). The mixture was heated
at 50.degree. C. to be completely dissolved, and evaporated to
dryness under a reduced pressure. Acetonitrile was added, and the
resulting mixture was again evaporated to dryness under a reduced
pressure. Acetone was added to the residue, and the resulting
mixture was ultrasonically treated and filtered. The filter cake
was dried to produce 40 mg of a yellow solid with a yield of
61%.
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.99 (s, 1H), 9.88 (s, 1H),
8.63 (s, 2H), 8.40 (s, 1H), 8.28 (s, 1H), 8.19 (s, 1H), 7.52 (d,
J=7.3 Hz, 1H), 7.25 (s, 1H), 7.10 (s, 1H), 6.88-6.70 (m, 1H), 6.26
(d, J=16.8 Hz, 1H), 5.75 (d, J=8.6 Hz, 1H), 5.18 (br s, 1H), 3.92
(s, 3H), 3.46 (s, 2H), 3.31 (s, 2H), 2.79 (s, 9H), 2.38 (s, 3H),
1.32-1.12 (m, 6H).
Example 17:
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{5-chloro-[-
4-(1-methyl-5,6-difluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}a-
crylamide methanesulfonate
##STR00100##
The compound was synthesized in the substantially same manner as
those in Example 16. Ethyl acetate was added to the final crude
product. The mixture was ultrasonically treated and filtered to
produce a product with a yield of 43%.
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 10.02 (s, 1H), 9.95 (s, 1H),
8.90 (s, 1H), 8.71 (s, 1H), 8.39 (s, 1H), 8.21 (s, 1H), 8.09 (s,
1H), 7.69 (dd, J=11.1, 7.0 Hz, 1H), 6.84 (dd, J=17.0, 10.2 Hz, 1H),
6.23 (dd, J=17.1, 1.7 Hz, 1H), 5.73 (dd, J=10.3, 1.7 Hz, 1H), 5.17
(hept, J=6.1 Hz, 1H), 3.90 (s, 3H), 3.61 (t, J=5.6 Hz, 2H), 3.32
(d, J=5.5 Hz, 2H), 2.79 (s, 6H), 2.78 (s, 3H), 2.39 (s, 3H), 1.18
(d, J=6.1 Hz, 6H).
Example 18:
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[4-(1-meth-
yl-5,6-difluoro-1H-indol-3-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylamide
methanesulfonate
##STR00101##
The compound was synthesized in the substantially same manner as
those in Example 16. Ethyl acetate was added to the final crude
product. The mixture was ultrasonically treated and filtered to
produce a product with a yield of 96%.
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 9.99 (s, 2H), 8.82 (s, 1H),
8.26-8.11 (m, 3H), 7.81 (dd, J=10.6, 6.9 Hz, 1H), 7.40 (d, J=6.6
Hz, 1H), 6.82 (dd, J=16.9, 10.3 Hz, 11H), 6.27 (d, J=17.0 Hz, 1H),
5.78 (d, J=10.1 Hz, 1H), 5.25-5.19 (m, 1H), 3.91 (s, 3H), 3.68 (d,
J=5.5 Hz, 2H), 3.35 (d, J=5.5 Hz, 2H), 2.86 (s, 3H), 2.82 (s, 3H),
2.80 (s, 3H), 2.36 (s, 3H), 1.21 (d, J=6.1 Hz, 6H).
Example 19:
.[.N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[5-chlo-
ro-4-(1-methyl-1H-pyrro[2,3-b]pyridin-3-yl)pyrimidin-2-yl]amino}pyridin-3--
yl}acrylamide.].
.Iadd.N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[5-c-
hloro-4-(1-methyl-1H-pyrrolo[2,3-b]pyridin-3-yl)pyrimidin-2-yl]amino}pyrid-
in-3-yl}acrylamide.Iaddend.
##STR00102##
Step 1: Synthesis of
.[.N.sup.2-methyl-N.sup.2-[2-(dimethylamino)ethyl]-6-isopropyloxy-N.sup.5-
-[5-chloro-4-(1-methyl-1H-pyrro[2,3-b]pyridin-3-yl)pyrimidin-2-yl]-3-nitro-
pyridin-2,5-diamine.].
.Iadd.N.sup.2-methyl-N.sup.2-[2-(dimethylamino)ethyl]-6-isopropyloxy-N.su-
p.5-[5-chloro-4-(1-methyl-1H-pyrrolo[2,3-b]pyridin-3-yl)pyrimidin-2-yl]-3--
nitropyridin-2,5-diamine.Iaddend.
##STR00103##
The compound was synthesized in the same manner as those in Step 1
of Example 1 with a yield of 46%. MS m/z: 540.
Step 2: Synthesis of
.[.N.sup.2-methyl-N.sup.2-[2-(dimethylamino)ethyl]-6-isopropyloxy-N.sup.5-
-[5-chloro-4-(1-methyl-1H-pyrro[2,3-b]pyridin-3-yl)pyrimidin-2-yl]pyridin--
2,3,5-triamine.].
.Iadd.N.sup.2-methyl-N.sup.2-[2-(dimethylamino)ethyl]-6-isopropyloxy-N.su-
p.5-[5-chloro-4-(1-methyl-1H-pyrrolo[2,3-b]pyridin-3-yl)pyrimidin-2-yl]pyr-
idin-2,3,5-triamine.Iaddend.
##STR00104##
The compound was synthesized in the same manner as those in Step 2
of Example 1 with a yield of 37%. MS m/z: 510 [M+1].
Step 3: Synthesis of
.[.N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[5-chlo-
ro-4-(1-methyl-1H-pyrro[2,3-b]pyridin-3-yl)pyrimidin-2-yl]amino}pyridin-3--
yl}acrylamide.].
.Iadd.N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[5-c-
hloro-4-(1-methyl-1H-pyrrolo[2,3-b]pyridin-3-yl)pyrimidin-2-yl]amino}pyrid-
in-3-yl}acrylamide.Iaddend.
##STR00105##
The compound was synthesized in the same manner as those in Step 3
of Example 1 with a yield of 52%. MS m/z: 564 [M+1].
.sup.1H NMR (400 MHz, MeOD) .delta. 8.70 (dd, J=8.0, 1.0 Hz, 1H),
8.61 (s, 1H), 8.41 (s, 1H), 8.40 (s, 1H), 8.29 (dd, J=4.7, 1.5 Hz,
1H), 7.14 (dd, J=8.0, 4.8 Hz, 1H), 6.48 (dd, J=16.9, 2.6 Hz, 1H),
6.42 (dd, J=16.9, 9.2 Hz, 1H), 5.86 (dd, J=9.2, 2.6 Hz, 1H),
5.38-5.32 (m, 1H), 3.97 (s, 3H), 3.74 (t, J=5.7 Hz, 2H), 3.33 (t,
J=5.7 Hz, 2H), 2.90 (s, 6H), 2.80 (s, 3H), 1.41 (d, J=6.2 Hz,
6H).
Example 20:
.[.N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[5-chlo-
ro-4-(1-methyl-1H-pyrro[2,3-b]pyridin-5-yl)pyrimidin-2-yl]amino}pyridin-3--
yl}acrylamide.].
.Iadd.N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[5-c-
hloro-4-(1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)pyrimidin-2-yl]amino}pyrid-
in-3-yl}acrylamide.Iaddend.
##STR00106##
The compound was prepared in the same manner as those in Example 7.
MS m/z: 564 [M+1].
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.69 (s, 1H), 9.55 (s,
1H), 8.99 (d, J=1.9 Hz, 1H), 8.71 (s, 1H), 8.50 (s, 1H), 7.52 (s,
1H), 7.24 (d, J=3.5 Hz, 1H), 7.14 (dd, J=16.9, 10.2 Hz, 1H), 6.63
(d, J=3.5 Hz, 1H), 6.54 (dd, J=16.9, 1.9 Hz, 1H), 5.77 (dd, J=10.2,
1.9 Hz, 1H), 5.26 (hept, J=6.2 Hz, 1H), 3.94 (s, 3H), 3.52 (t,
J=5.2 Hz, 2H), 3.11 (t, J=5.2 Hz, 2H), 2.81 (s, 3H), 2.79 (s, 3H),
1.38 (d, J=6.2 Hz, 6H).
Example 21:
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[5-chloro--
4-(1-methyl-1H-pyrazol-4-yl)pyrimidin-2-yl]amino}pyridin-3-yl}acrylamide
##STR00107##
The compound was prepared in the same manner as those in Example 7
with a yield of 8%. MS m/z: 514 [M+1].
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.79 (s, 1H), 9.55 (s,
1H), 9.12 (s, 1H), 8.46 (s, 1H), 8.33 (s, 1H), 7.48 (s, 1H), 6.50
(dd, J=17.0, 2.0 Hz, 1H), 5.77 (dd, J=10.0, 2.0 Hz, 1H), 5.30 5.22
(m, 1H), 4.08 (s, 3H), 3.57 (s, 2H), 3.16 (s, 2H), 2.83 (s, 9H),
1.39 (d, J=6.2 Hz, 6H).
Example 22:
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[5-chloro--
2'-methoxy-(4,5'-bipyrimidine)-2-yl]amino}pyridin-3-yl}acrylamide
##STR00108##
The compound was synthesized in the same manner as those in Example
7.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.75 (s, 1H), 9.50 (s,
1H), 9.27 (s, 2H), 8.51 (s, 1H), 7.52 (s, 1H), 7.17-7.03 (m, 1H),
6.57 (d, J=16.9 Hz, 1H), 5.76 (d, J=12.0 Hz, 1H), 5.31-5.23 (m,
1H), 4.13 (s, 3H), 3.54 (s, 2H), 3.12 (s, 2H), 2.83 (s, 3H), 2.81
(s, 6H), 1.40 (d, J=6.2 Hz, 6H).
Example 23:
N-{2-{[2-(dimethylamino)ethyl](methyl)amino}-6-isopropyloxy-5-{[5-chloro--
2'-amino-(4,5'-bipyrimidine)-2-yl]amino}pyridin-3-yl}acrylamide
##STR00109##
The compound was prepared in the same manner as those in Example 7
with a yield of 8%. MS m/z: 527 [M+1].
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.76 (s, 1H), 9.43 (s,
1H), 9.09 (s, 2H), 8.45 (s, 1H), 7.48 (s, 1H), 7.02 (s, 1H), 6.52
(dd, J=16.9, 1.8 Hz, 1H), 5.75 (dd, J=10.3, 1.8 Hz, 1H), 5.61 (s,
2H), 5.26 (hept, J=6.2 Hz, 1H), 3.47 (br s, 2H), 3.05 (br s, 2H),
2.81 (s, 3H), 2.76 (s, 6H), 1.39 (d, J=6.2 Hz, 6H).
II. Examples of the Activity Test of the Present Compounds
Test Example 1: Proliferation Inhibition Effects on Human Skin
Cancer Cell (A431, Wild-Type EGFR), Human Lung Cancer Cell (HCC827,
EGFR Exon 19 Deletion Activating Mutation), Human Lung Cancer Cell
(H1975, EGFR L858R/T790M Resistant Mutation)
Cells in the logarithmic phase were inoculated to 96-well culture
plates (cell density: 5000/well, cell suspension: 180 .mu.l/well),
and cultured at 37.degree. C. under 5% CO.sub.2 for 24 hours. After
the culturing, the cells adhered to the well walls. Each of
compounds was dissolved in DMSO in advance to formulate a 10 nM
stock solution. Upon testing, the stock solution was diluted with
complete medium to 10 times the target concentration in another
96-cell plate. And then the compound was added at 20 .mu.l/cell in
the 96-well plate in which the cells were inoculated, i.e. the
target concentration could be reached. The well for each
concentration was triplicated, and the blank control was
established. Cells continued to be cultured at 37.degree. C. under
5% CO.sub.2 for 72 hours. After the termination of culturing, 50
.mu.l pre-cooled (4.degree. C.) 50% trichloroacetic acid, i.e., TCA
was added to each of wells (final concentration=10%), and was
placed at 4.degree. C. for 1 hour to fix the cells. The resulting
matter was washed with purified water for at least 5 times, and
dried naturally in air or at 60.degree. C. in an oven. 4 mg/ml
Sulforhodamine B (SRB) solution prepared by 1% glacial acetic
acid/purified water was added at 100 .mu.l/well to each well so as
to stain for 1 hour at room temperature. The supernatant was
discarded. The residue was washed with 1% acetic acid for at least
5 times to remove the non-specifically binding, and dried for use.
To each well was added 150 .mu.l of 10 mM Tris-HCl solution for
dissolving the contents therein. The OD value was measured at a
wavelength of 510 nm, and the inhibition rate was calculated based
on the collected data. The result was shown in Table 1.
TABLE-US-00002 TABLE 1 HCC827 H1975 A431 IC.sub.50 (nM) IC.sub.50
(nM) IC.sub.50 (nM) AZD9291 3.80 5.43 70.43 Example 1 compound 2.15
5.64 140.5 Example 2 compound 4.22 5.54 195.5 Example 3 compound
1.34 2,28 224.2 Example 4 compound 1.92 6.15 163.7 Example 5
compound 2.58 4.83 181.4 Example 6 compound 2.36 5.20 337.8 Example
7 compound 1.40 16.68 307.0 Example 8 compound 5.98 8.40 375.4
Example 9 compound 1.17 12.58 697.2 Example 10 compound 2.62 5.32
208.9 Example 11 compound 2.23 7.22 210.3 Example 12 compound 0.96
9.01 338.6 Example 13 compound 2.62 5.33 208.9 Example 14 compound
0.77 5.17 241.8 Example 15 compound 0.69 6.28 337.4 Example 16
compound 1.45 5.43 273.4 Example 17 compound 5.56 7.63 375.4
Example 18 compound 2.24 5.07 341.3 Example 19 compound 2.62 2.56
208.9 Example 20 compound 8.97 42.35 800.7 Example 21 compound
142.4 18.55 369.8 Example 22 compound 33.30 37.98 2765 Example 23
compound 3.08 30.70 1145 Note: AZD9291 was prepared according to
Example 28 of WO 2013/014448 A1
The test results showed that the compounds of the present invention
had a strong proliferation inhibition effect on human lung cancer
cell (HCC827, EGFR Exon 19 deletion activating mutation) and human
lung cancer cell (H1975, EGFR L858R/T790M resistant mutation), a
relatively weak proliferation inhibition effect on human skin
cancer cell (A431, wild-type EGFR), that is to say, the compounds
of the present invention had a good selectivity.
Test Example 2: Inhibition Effect on the Growth of Subcutaneously
Transplanted Tumors of Human Lung Cancer H1975-Bearing Nude
Mice
The Inhibition effect of the compound of Example 3 of the present
invention and AZD9291 on subcutaneously transplanted tumors of
human lung cancer H1975-bearing nude mice and the corresponding
safety were observed.
Cell cultivation: H1975 was placed in a RPMI-1640 medium containing
10% FBS, and cultivated in a temperature-constant incubator
containing 5% CO.sub.2 at 37.degree. C. The cells in exponential
growth phase were collected and counted for inoculation.
Test animals: 15 BALB/c nude mices, 15 males and 0 female, 6 weeks
old, 18-20 g, commercially available from Shanghai Lab. Animal
Research Center
Three test groups were established: 0.5% sodium
carboxymethylcellucose solvent control group, the groups of the
compound of Example 3 at 25 mg/kg and the groups of AZD9291 at 25
mg/kg, respectively.
Experimental method: human lung cancer H1975 cell strain
(5.times.10.sup.6/each mouse) was inoculated to nude mice
subcutaneously at the right side of the back thereof. Each mouse
was inoculated with 0.1 ml, and the tumor growth was observed
regularly. After the tumors grew to 100-150 mm.sup.3 on average,
the mice were divided into groups randomly according to the tumor
size and the mouse weight. The compound of Example 3 and AZD9291
were administered by intragastric administration in the dosage of
25 mg/kg, and solvent control groups were administered with equal
amount of solvent by intragastric administration, wherein the
administration was performed once per day for a continuous period
of 12 days. During the entire experimental process, the mouse
weight and the tumor size were measured twice per week, so as to
observe whether or not the toxic reaction occurs. The tumor volume
is calculated as follows: Tumor volume (mm.sup.3)=0.5.times.(Tumor
major diameter.times.Tumor minor diameter.sup.2)
The tumor growth curves of three experimental groups are shown in
FIG. 1, and the mice's weight growth curves are shown in FIG. 2.
The results show that the compounds of the present invention have a
good inhibition effect on the growth of subcutaneously transplanted
tumors of human lung cancer H1975-bearing nude mice, while having
little effect on the weights of nude mice, and showing a good
safety.
All of the literatures mentioned herein are incorporated into the
present application by reference. It should be also noted that,
upon reading the above mentioned contents of the present
application, a person skilled in the art can modify, change or
amend the present invention without departing from the spirits of
the present invention, and these equivalents are also within the
scope as defined by the claims appended in the present
application.
* * * * *