U.S. patent application number 14/950347 was filed with the patent office on 2016-03-17 for heterocyclic substituted-3-heteroarylidenyl-2-indolinone derivative.
The applicant listed for this patent is BOSTON BIOMEDICAL, INC.. Invention is credited to Toshio Kanai, Tomohiro Kodama, Katsumi Kubota, Yosuke Takanashi, Shingo Tojo, Katsunori Tsuboi.
Application Number | 20160075697 14/950347 |
Document ID | / |
Family ID | 51529945 |
Filed Date | 2016-03-17 |
United States Patent
Application |
20160075697 |
Kind Code |
A1 |
Tsuboi; Katsunori ; et
al. |
March 17, 2016 |
HETEROCYCLIC SUBSTITUTED-3-HETEROARYLIDENYL-2-INDOLINONE
DERIVATIVE
Abstract
Disclosed is a compound represented by formula (1) or a
pharmacologically acceptable salt thereof. (In the formula, R.sub.1
is optionally substituted heteroaryl etc.; R.sub.2 is hydrogen
etc.; R.sub.3 and R.sub.4 are each independently hydrogen etc.,
R.sub.5 is the following group: (wherein Y is optionally
substituted five membered heteroaryl etc., R.sub.9a is optionally
substituted aryl etc., R.sub.9b and R.sub.9c are each dependently
hydrogen etc., and m is the integral 0 etc.) etc.; R.sub.6 is
hydrogen etc.; and R.sub.7 is hydrogen etc. ##STR00001##
Inventors: |
Tsuboi; Katsunori; (Osaka,
JP) ; Takanashi; Yosuke; (Osaka, JP) ; Tojo;
Shingo; (Osaka, JP) ; Kodama; Tomohiro;
(Osaka, JP) ; Kubota; Katsumi; (Osaka, JP)
; Kanai; Toshio; (Osaka, JP) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
BOSTON BIOMEDICAL, INC. |
Cambridge |
MA |
US |
|
|
Family ID: |
51529945 |
Appl. No.: |
14/950347 |
Filed: |
November 24, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14208289 |
Mar 13, 2014 |
9227962 |
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14950347 |
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61780263 |
Mar 13, 2013 |
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Current U.S.
Class: |
544/131 ;
544/140; 544/144; 544/331; 544/365; 544/367; 544/371; 544/373;
546/201; 546/277.7; 548/136; 548/145; 548/204; 548/236; 548/255;
548/266.4; 548/364.7; 548/466 |
Current CPC
Class: |
C07D 403/14 20130101;
C07D 417/14 20130101; C07D 409/14 20130101; C07D 413/14 20130101;
C07D 401/14 20130101; C07D 405/14 20130101 |
International
Class: |
C07D 417/14 20060101
C07D417/14; C07D 413/14 20060101 C07D413/14; C07D 405/14 20060101
C07D405/14; C07D 409/14 20060101 C07D409/14; C07D 403/14 20060101
C07D403/14 |
Claims
1. A compound of formula 1: ##STR00154## or pharmaceutically
acceptable salt thereof, R.sub.1 is optionally substituted
heteroaryl, or optionally substituted heteroalicyclic; R.sub.2 is
hydrogen, optionally substituted alkyl, optionally substituted
alkylcarbonyl, optionally substituted alkoxycarbonyl, or optionally
substituted aminocarbonyl; R.sub.3 and R.sub.4 are each
independently hydrogen, halogen, cyano, nitro, hydroxy, optionally
substituted alkyl, optionally substituted alkoxy, optionally
substituted amino, optionally substituted alkoxycarbonyl,
optionally substituted alkylcarbonyl, optionally substituted
aminocarbonyl, or the following group: ##STR00155## (wherein X is
optionally substituted five membered heteroaryl or optionally
substituted heteroalicyclic, R.sub.8a is optionally substituted
aryl, optionally substituted heteroaryl, or optionally substituted
heteroalicyclic, R.sub.8b and R.sub.8c are each independently
hydrogen, alkyl, halogen, cyano, amino, nitro, hydroxy,
trifluoromethyl, optionally substituted aryl, optionally
substituted heteroaryl, or optionally substituted heteroalicyclic,
and n is the integral 0 to 2), R.sub.5 is hydrogen, halogen, cyano,
nitro, hydroxy, optionally substituted alkyl, optionally
substituted alkoxy, optionally substituted amino, optionally
substituted alkoxycarbonyl, optionally substituted alkylcarbonyl,
optionally substituted aminocarbonyl, or the following group:
##STR00156## (wherein Y is optionally substituted five membered
heteroaryl (provided that the five membered heteroaryl is not the
following group: ##STR00157## wherein R.sub.9a is same as the above
definition), R.sub.9a is optionally substituted aryl, optionally
substituted heteroaryl, or optionally substituted heteroalicyclic,
R.sub.9b and R.sub.9c are each dependently hydrogen, alkyl,
halogen, cyano, amino, nitro, hydroxy, trifluoromethyl, and m is
the integral 0 to 2), and at least one of R.sub.3, R.sub.4 and
R.sub.5 is the following group: ##STR00158## (X, R.sub.8a,
R.sub.8b, R.sub.8c and n are same as the above definition), or the
following group: ##STR00159## (Y, R.sub.9a, R.sub.9b, R.sub.9c and
m are same as the above definition); R.sub.6 is hydrogen, halogen,
cyano, nitro, hydroxy, amino, optionally substituted alkyl,
optionally substituted alkoxy, optionally substituted amino,
optionally substituted alkoxycarbonyl, optionally substituted alkyl
carbonyl, or optionally substituted aminocarbonyl; and R.sub.7 is
hydrogen, optionally substituted alkyl, or optionally substituted
cycloalkyl.
2. The compound according to claim 1, or pharmaceutically
acceptable salt thereof, R.sub.1 is optionally substituted
heteroaryl, or optionally substituted heteroalicyclic, wherein the
heteroaryl is selected from the group consisting of pyrrole, furan,
thiophene, pyridine, pyrimidine, pyrazine, pyridazine, indole, and
pyrrolopyridinone, and heteroalicyclic is selected from the group
consisting of pyridone, pyrrolidine, and piperidine.
3. The compound according to claim 2, or pharmaceutically
acceptable salt thereof, R.sub.1 is optionally substituted
heteroaryl, or optionally substituted heteroalicyclic, wherein
heteroaryl is pyrrole, or pyrrolopyridinone, and heteroalicyclic is
pyridone.
4. The compound according to anyone of claims 1-3, or
pharmaceutically acceptable salt thereof, the substituent of
optionally substituted heteroaryl and optionally substituted
heteroalicyclic in R.sub.1 is one or more substituent(s) selected
from the group consisting of halogen, alkyl, optionally substituted
alkoxy, optionally substituted amino, optionally substituted alkoxy
carbonyl, optionally substituted alkylcarbonyl, or optionally
substituted aminocarbonyl.
5. The compound according to anyone of claims 1-4, or
pharmaceutically acceptable salt thereof, R.sub.1 is one of the
group selected from the following group: ##STR00160## (wherein
R.sub.10 is hydroxy, optionally substituted alkyl, optionally
substituted alkoxy, or optionally substituted amino).
6. The compound according to claim 5, or pharmaceutically
acceptable salt thereof, R.sub.10 is hydroxy, mono-substituted
alkyl amino (said alkyl is substituted by di-substituted alkyl
amino, or heteroalicyclic), piperazynyl, homopiperazynyl or
morphorinyl.
7. The compound according to claim 6, or pharmaceutically
acceptable salt thereof, R.sub.10 is hydroxy, ethyl amino (said
ethyl is substituted by di-ethyl amino, 1-pyrrolidino), or
4-piperazinyl (said piperazinyl is substituted with alkyl).
8. The compound according to claim 7, or pharmaceutically
acceptable salt thereof, R.sub.10 is hydroxy, 2-(di-ethyl
amino)ethyl amino, 2-pyrrolidino ethyl amino, 4-methyl piperazinyl
or morpholino.
9. The compound according to anyone of claims 1-8, or
pharmaceutically acceptable salt thereof, R.sub.2 is hydrogen, or
optionally substituted alkoxy carbonyl.
10. The compound according to claim 9, or pharmaceutically
acceptable salt thereof, R.sub.2 is hydrogen.
11. The compound according to anyone of claims 1-10, or
pharmaceutically salt thereof, R.sub.3 and R.sub.4 are each
independently 1: hydrogen, 2: halogen, 3: cyano, 4: nitro, 5:
hydroxy, 6: alkyl. 7: alkoxy, 8: amino, 9: alkylcarbonyl, 10:
alkoxycarbonyl, 11: aminocarbonyl, and 12: the following group:
##STR00161## (wherein X, R.sub.8a, R.sub.8b, R.sub.8c, and n are
same as the above definition), wherein said alkyl, alkoxy, amino,
alkylcarbonyl, alkoxycarbonyl, and aminocarbonyl are optionally
substituted with one or more substitutent(s) selected from the
group consisting of (a) alkyl, (b) alkenyl, (c) alkynyl, (d)
hydroxy, (e) amino, (f) nitro, (g) cyano, (h) halogen, (i) alkoxy,
(j) alkylcarbonyl, (k) alkoxycarbonyl, (l) aminocarbonyl, (m) aryl,
(n) heteroaryl, (o) cycloalkyl, and (p) heteroalicyclic).
12. The compound according to claim 11, or pharmaceutically
acceptable salt thereof, R.sub.3 and R.sub.4 are each independently
1: hydrogen, 2: halogen, 3: alkyl(said alkyl is optionally
substituted with one or more substituent(s) selected from the group
consisting of (a) alkyl, (b) alkenyl, (c) alkynyl, (d) hydroxy, (e)
amino, (f) nitro, (g) cyano, (h) halogen, (i) alkoxy, (j)
alkylcarbonyl, (k) alkoxycarbonyl, (l) aminocarbonyl, (m) aryl, (n)
heteroaryl, (o) cycloalkyl, and (p) heteroalicyclic) 4: the
following group: ##STR00162## (wherein X, R.sub.8a, R.sub.8b,
R.sub.8c, and n are same as the above definition).
13. The compound according to claim 12, or pharmaceutically
acceptable salt thereof, R.sub.3 and R.sub.4 are each independently
hydrogen, or the following group: ##STR00163## (wherein X,
R.sub.8a, R.sub.8b, R.sub.8c, and n are same as the above
definition).
14. The compound according to anyone of claims 1-13, or
pharmaceutically acceptable salt thereof, X is pyrrole, furan,
thiophene, imidazole, pyrazole, oxazole, isoxazole, thiazole,
isothiazole, triazole, thiadiazole, oxadiazole, pyrrolidine,
piperidine, azepane, tetrahydrofuran, oxane, or oxepane.
15. The compound according to claim 14, or pharmaceutically
acceptable salt thereof, X is pyrrole, furan, thiophene, imidazole,
pyrazole, oxazole, isoxazole, thiazole, isothiazole, triazole,
thiadiazole, or oxadiazole.
16. The compound according to claim 15, or pharmaceutically
acceptable salt thereof, X is thiophene, pyrazole, oxazole,
thiazole, thiadiazole, or oxadiazole.
17. The compound according to claim 16, or pharmaceutically
acceptable salt thereof, X is pyrazole, thiazole, or oxazole.
18. The compound according to claim 1, or pharmaceutically
acceptable salt thereof, R.sub.8a, R.sub.8b and R.sub.8c are each
independently hydrogen, alkyl, halogen, cyano, amino, nitro,
hydroxy, trifluoromethyl, phenyl, or pyridyl.
19. The compound according to claim 18, or pharmaceutically
acceptable salt thereof, R.sub.8a, R.sub.8b and R.sub.8c are each
dependently hydrogen, alkyl, phenyl, or pyridyl.
20. The compound according to claim 1, or pharmaceutically
acceptable salt thereof, n is the integra 0.
21. The compound according to claim 1, or pharmaceutically
acceptable salt thereof, X is selected from the group consisting of
the following group: ##STR00164## R.sub.8a is piperidinyl, pyranyl,
phenyl, thiophenyl, pyrazinyl, pyrimidinyl, pyridazinyl, or pyridyl
(said piperidinyl, pyranyl, phenyl, thiophenyl pyrazinyl,
pyrimidinyl, pyridazinyl, or pyridyl are optionally substituted
with halogen, hydroxy, cyano, amino, nitro, alkyl, alkoxy,
alkoxycarbonyl, alkylcarbonyl, or aminocarbonyl), and R.sub.8b is
alkyl.
22. The compound according to claim 1, or pharmaceutically
acceptable salt thereof, R.sub.3 and R.sub.4 are hydrogen.
23. The compound according to claim 1, or pharmaceutically
acceptable salt thereof, R.sub.5 is selected from the group
consisting of the following group: ##STR00165## R.sub.9a
piperidinyl, pyranyl, phenyl, thiophenyl, pyrazinyl, pyrimidinyl,
pyridazinyl, or pyridyl (said piperidinyl, pyranyl, phenyl,
thiophenyl, pyrazinyl, pyrimidinyl, pyridazinyl, or pyridyl are
optionally substituted with halogen, hydroxy, cyano, amino, nitro,
alkyl, alkoxy, alkoxycarbonyl, alkylcarbonyl, or aminocarbonyl).
and R.sub.9b is alkyl.
24. The compound according to claim 1, or pharmaceutically
acceptable salt thereof, R.sub.6 is hydrogen, halogen, methyl,
ethyl, n-propyl, isopropyl, trifluoromethyl, optionally substituted
alkoxy, optionally substituted amino, optionally substituted
alkoxycarbonyl, optionally substituted alkylcarbonyl, or optionally
substituted aminocarbonyl.
25. The compound according to claim 1, or pharmaceutically
acceptable salt thereof, R.sub.7 is hydrogen, methyl, ethyl,
n-propyl, or isopropyl.
26. A pharmaceutical composition comprising the compound of claim
1, or pharmaceutically acceptable salt thereof.
Description
TECHNICAL FIELD
[0001] The present invention relates to novel heterocyclic
substituted-3-heteroarylidenyl-2-indolinone derivative, or a
pharmaceutically acceptable salts thereof which modulate the
activity of protein kinases ("PKs"). The present invention also
relates to pharmaceutical composition comprising heterocyclic
substituted-3-heteroarylidenyl-2-indolinone derivative, or a
pharmaceutically acceptable salts thereof. The present invention
also relates to a therapeutic or preventive agent or method for
treating disorders related to abnormal PK activity.
BACKGROUND ART
[0002] The protein kinases represent a large family of proteins,
which plays a central role of in the regulation of wide variety of
cellular processes and maintaining control over cellular function
as enzymes that catalyze the phosphorylation of hydroxyl groups on
tyrosine, serine and threonine residues of protein. A partial,
non-limiting, list of these kinases include: receptor tyrosine
kinases such as platelet-derived growth factor receptor kinase
(PDGFR), insulin-like growth factor receptor (IGFR), the nerve
growth factor receptor, TrkB, Met, and the fibroblast growth factor
receptor, FGFR-3; non-receptor tyrosine kinase such as Abl and the
function kinase Bcr-Abl, Lck, Csk, Fes, Bmx, and Src; and
serine/threonint kinases such as B-Raf, C-Raf, Sgk, MAP kinases
(e.g., MKK4, MKK6, etc.) and SAPK2a, SAPK2b and SAPK3. The
consequences of phosphorylation with PKs are staggering; cell
growth, differentiation and proliferation. Furthermore, aberrant
kinase activity has been observed in many disease states including
benign and malignant proliferative disorders, as well as diseases
resulting from inappropriate activation of the immune and nerve
systems. In view of the apparent link between PK-related cellular
activities and wide variety of human disorders, a great deal of
effort is being expended in an attempt to identify ways to modulate
PK activity.
[0003] Some of this effort has involved biomimetic approaches using
large molecules patterned on those involved in the actual cellular
processes (mutant ligands (Patent Document 1); soluble receptors
and antibodies (Patent Document 2) and tyrosine kinase inhibitors
(Patent Document 3.).
[0004] In addition to the above, attempts have been made to
identify small molecules which act as PK inhibitors, For example,
bis-monocyclic, bicyclic and heterocyclic aryl compounds (Patent
Document 4), vinylene azaindole derivatives (Patent Document 5)
have been described as tyrosine kinase inhibitors. However these
compounds have limited utility because of toxicity, poor
bioavailability, or less potency.
[0005] On the other hand, some of indolinone derivatides are
reported as PK inhibitors (Patent Document 6). Especially,
4-phenyl- or 4-pyridyl indolinone derivatives (Patent Document 7),
and 5-(thiazol-4-yl)indolin-2-one derivatives (Patent Document 8)
are reported.
[0006] However these prior arts do not disclose derivatives with
other five-membered heteroaryl groups or heteroalicyclic group at
5.sup.th, 6.sup.th, or 7.sup.th position on indolinone and
indolin.
[0007] [Patent Document 1] U.S. Pat. No. 4,966,849
[0008] [Patent Document 2] WO94/10202 pamphlet
[0009] [Patent Document 3] WO92/21660 pamphlet
[0010] [Patent Document 4] WO92/20642 pamphlet
[0011] [Patent Document 5] WO94/14808 pamphlet
[0012] [Patent Document 6] U.S. Pat. No. 5,792,783
[0013] [Patent Document 7] WO02/02551 pamphlet
[0014] [Patent Document 8] WO2009/033033 pamphlet
SUMMARY OF INVENTION
Problems to be Solved by the Invention
[0015] Recently, a pharmaceutically satisfiable compound inhibiting
CSCPKs has been desired as an agent for treating diseases including
cancer etc.
[0016] According to the extensive studies for solving the problem,
the inventors have found the following heterocyclic
substituted-3-heteroarylidenyl-2-indolinone derivatives
structurally characterized by specific group substituted the
five-membered heteroaryl groups or heteroalicyclic group at
5.sup.th, 6.sup.th, or 7.sup.th position on indolinone have potent
CSCPKs inhibitory activity.
[0017] The inventors have also found that the derivatives have
suitable properties essential for a medicament, including
solubility, membrane permeability, metabolic stability,
bioavailability, safety for heart (such as hERG), as well as CSCPKs
inhibitory activity, and achieved the present invention.
Means of Solving the Problems
[0018] Specifically, the present invention is as follows.
[0019] Item1. A compound of formula 1:
##STR00002##
or pharmaceutically acceptable salt thereof, R.sub.1 is optionally
substituted heteroaryl, or optionally substituted heteroalicyclic;
R.sub.2 is hydrogen, optionally substituted alkyl, optionally
substituted alkylcarbonyl, optionally substituted alkoxycarbonyl,
or optionally substituted aminocarbonyl; R.sub.3 and R.sub.4 are
each independently hydrogen, halogen, cyano, nitro, hydroxy,
optionally substituted alkyl, optionally substituted alkoxy,
optionally substituted amino, optionally substituted
alkoxycarbonyl, optionally substituted alkyl carbonyl, optionally
substituted aminocarbonyl, or the following group:
##STR00003##
(wherein X is optionally substituted five membered heteroaryl or
optionally substituted heteroalicyclic, R.sub.8a is optionally
substituted aryl, optionally substituted heteroaryl, or optionally
substituted heteroalicyclic, R.sub.8b and R.sub.8c are each
independently hydrogen, alkyl, halogen, cyano, amino, nitro,
hydroxy, trifluoromethyl, optionally substituted aryl, optionally
substituted heteroaryl, or optionally substituted heteroalicyclic,
and n is the integral 0 to 2), R.sub.5 is hydrogen, halogen, cyano,
nitro, hydroxy, optionally substituted alkyl, optionally
substituted alkoxy, optionally substituted amino, optionally
substituted alkoxycarbonyl, optionally substituted alkylcarbonyl,
optionally substituted aminocarbonyl, or the following group:
##STR00004##
(wherein Y is optionally substituted five membered
heteroaryl(provided that the five membered heteroaryl is not the
following group:
##STR00005##
wherein R.sub.9a is same as the above definition), R.sub.9a is
optionally substituted aryl, optionally substituted heteroaryl, or
optionally substituted heteroalicyclic, R.sub.9b and R.sub.9c are
each dependently hydrogen, alkyl, halogen, cyano, amino, nitro,
hydroxy, trifluoromethyl, and m is the integral 0 to 2), and at
least one of R.sub.3, R.sub.4 and R.sub.5 is the following
group:
##STR00006##
(X, R.sub.8a, R.sub.8b, R.sub.8c and n are same as the above
definition), or the following group:
##STR00007##
(Y, R.sub.9a, R.sub.9b, R.sub.9c and m are same as the above
definition); R.sub.6 is hydrogen, halogen, cyano, nitro, hydroxy,
amino, optionally substituted alkyl, optionally substituted alkoxy,
optionally substituted amino, optionally substituted alkoxy
carbonyl, optionally substituted alkyl carbonyl, or optionally
substituted aminocarbonyl; and R.sub.7 is hydrogen, optionally
substituted alkyl, or optionally substituted cycloalkyl.
[0020] Item2. The compound according to Item 1, or pharmaceutically
acceptable salt thereof,
R.sub.1 is optionally substituted heteroaryl, or optionally
substituted heteroalicyclic, wherein the heteroaryl is selected
from the group consisting of pyrrole, furan, thiophene, pyridine,
pyrimidine, pyrazine, pyridazine, indole, and pyrrolopyridinone,
and heteroalicyclic is selected from the group consisting of
pyridone, pyrrolidine, and piperidine.
[0021] Item3. The compound according to Item 2, or pharmaceutically
acceptable salt thereof,
R.sub.1 is optionally substituted heteroaryl, or optionally
substituted heteroalicyclic, wherein heteroaryl is pyrrole, or
pyrrolopyridinone, and heteroalicyclic is pyridone.
[0022] Item4. The compound according to anyone of Items 1-3, or
pharmaceutically acceptable salt thereof,
the substituent of optionally substituted heteroaryl and optionally
substituted heteroalicyclic in R.sub.1 is one or more
substituent(s) selected from the group consisting of halogen,
alkyl, optionally substituted alkoxy, optionally substituted amino,
optionally substituted alkoxy carbonyl, optionally substituted
alkylcarbonyl, or optionally substituted aminocarbonyl.
[0023] Item5. The compound according to anyone of Items 1-4, or
pharmaceutically acceptable salt thereof,
R.sub.1 is one of the group selected from the following group:
##STR00008##
(wherein R.sub.10 is hydroxyl, optionally substituted alkyl,
optionally substituted alkoxy, or optionally substituted
amino).
[0024] Item6. The compound according to Item 5, or pharmaceutically
acceptable salt thereof,
R.sub.10 is hydroxy, mono-substituted alkyl amino (said alkyl is
substituted by di-substituted alkyl amino, or heteroalicyclic),
piperazynyl, homopiperazynyl, or morphorinyl.
[0025] Item7. The compound according to Item 6, or pharmaceutically
acceptable salt thereof,
R.sub.10 is hydroxy, ethyl amino (said ethyl is substituted by
di-ethyl amino, 1-pyrrolidino), or 4-piperazinyl (said piperazinyl
is substituted with alkyl).
[0026] Item8. The compound according to Item 7, or pharmaceutically
acceptable salt thereof,
R.sub.10 is hydroxy, 2-(di-ethyl amino)ethyl amino, 2-pyrrolidino
ethyl amino, 4-methyl piperazinyl, or morpholino.
[0027] Item9. The compound according to anyone of Items 1-8, or
pharmaceutically acceptable salt thereof,
R.sub.2 is hydrogen, or optionally substituted alkoxy carbonyl.
[0028] Item10. The compound according to Item 9, or
pharmaceutically acceptable salt thereof,
R.sub.2 is hydrogen.
[0029] Item11. The compound according to anyone of Items 1-10, or
pharmaceutically salt thereof,
R.sub.3 and R.sub.4 are each independently
[0030] 1: hydrogen,
[0031] 2: halogen,
[0032] 3: cyano,
[0033] 4: nitro,
[0034] 5: hydroxy,
[0035] 6: alkyl.
[0036] 7: alkoxy,
[0037] 8: amino,
[0038] 9: alkylcarbonyl,
[0039] 10: alkoxycarbonyl,
[0040] 11: aminocarbonyl, and
[0041] 12: the following group:
##STR00009##
(wherein X, R.sub.8a, R.sub.8b, R.sub.8c, and n are same as the
above definition), wherein said alkyl, alkoxy, amino,
alkylcarbonyl, alkoxycarbonyl, and aminocarbonyl are optionally
substituted with one or more substitutent(s) selected from the
group consisting of
[0042] (a) alkyl,
[0043] (b) alkenyl,
[0044] (c) alkynyl,
[0045] (d) hydroxy,
[0046] (e) amino,
[0047] (f) nitro,
[0048] (g) cyano,
[0049] (h) halogen,
[0050] (i) alkoxy,
[0051] (j) alkylcarbonyl,
[0052] (k) alkoxycarbonyl,
[0053] (l) aminocarbonyl,
[0054] (m) aryl,
[0055] (n) heteroaryl,
[0056] (o) cycloalkyl, and
[0057] (p) heteroalicyclic).
[0058] Item12. The compound according to Item 11, or
pharmaceutically acceptable salt thereof,
R.sub.3 and R.sub.4 are each independently
[0059] 1: hydrogen,
[0060] 2: halogen,
[0061] 3: alkyl(said alkyl is optionally substituted with one or
more sunstituent(s) selected from the group consisting of
(a) alkyl, (b) alkenyl, (c) alkynyl, (d) hydroxy, (e) amino, (f)
nitro, (g) cyano, (h) halogen, (i) alkoxy, (j) alkylcarbonyl, (k)
alkoxycarbonyl, (l) aminocarbonyl, (m) aryl, (n) heteroaryl, (o)
cycloalkyl, and (p) heteroalicyclic)
[0062] 4: the following group:
##STR00010##
(wherein X, R.sub.8a, R.sub.8b, R.sub.8c, and n are same as the
above definition).
[0063] Item13. The compound according to Item 12, or
pharmaceutically acceptable salt thereof,
R.sub.3 and R.sub.4 are each independently hydrogen, or the
following group:
##STR00011##
(wherein X, R.sub.8a, R.sub.8b, R.sub.8c, and n are same as the
above definition).
[0064] Item14. The compound according to anyone of Items 1-13, or
pharmaceutically acceptable salt thereof,
X is pyrrole, furan, thiophene, imidazole, pyrazole, oxazole,
isoxazole, thiazole, isothiazole, triazole, thiadiazole,
oxadiazole, pyrrolidine, piperidine, azepane, tetrahydrofuran,
oxane, or oxepane.
[0065] Item15. The compound according to Item 14, or
pharmaceutically acceptable salt thereof,
X is pyrrole, furan, thiophene, imidazole, pyrazole, oxazole,
isoxazole, thiazole, isothiazole, triazole, thiadiazole, or
oxadiazole.
[0066] Item16. The compound according to Item 15, or
pharmaceutically acceptable salt thereof,
X is thiophene, pyrazole, oxazole, thiazole, thiadiazole, or
oxadiazole.
[0067] Item17. The compound according to Item 16, or
pharmaceutically acceptable salt thereof,
X is pyrazole, thiazole, or oxazole.
[0068] Item18. The compound according to Item 14, or
pharmaceutically acceptable salt thereof,
X is piperidine.
[0069] Item19. The compound according to anyone of Items 1-18, or
pharmaceutically acceptable salt thereof,
R.sub.8a, R.sub.8b and R.sub.8c are each independently hydrogen,
alkyl, halogen, cyano, amino, nitro, hydroxy, trifluoromethyl,
phenyl, or pyridyl.
[0070] Item20. The compound according to Item 19, or
pharmaceutically acceptable salt thereof,
R.sub.8a, R.sub.8b and R.sub.8c are each dependently hydrogen,
alkyl, phenyl, or pyridyl.
[0071] Item21. The compound according to anyone of Items 1-20, or
pharmaceutically acceptable salt thereof,
n is the integra 0.
[0072] Item22. The compound according to anyone of Items 1-10, or
pharmaceutically acceptable salt thereof,
X is selected from the group consisting of the following group:
##STR00012##
R.sub.8a is piperidinyl, pyranyl, phenyl, thiophenyl, pyrazinyl,
pyrimidinyl, pyridazinyl, or pyridyl (said piperidinyl, pyranyl,
phenyl, thiophenyl, pyrazinyl, pyrimidinyl, pyridazinyl, or pyridyl
are optionally substituted with halogen, hydroxy, cyano, amino,
nitro, alkyl, alkoxy, alkoxycarbonyl, alkylcarbonyl, or
aminocarbonyl), and R.sub.8b is alkyl.
[0073] Item23. The compound according to anyone of Items 1-10, or
pharmaceutically acceptable salt thereof,
X is selected from the group consisting of the following group:
##STR00013##
and R.sub.8a is 4-piperazinyl (said piperidinyl is substituted with
alkyl or alkylcarbonyl), 4-tetrahydropyranyl, phenyl, pyrazinyl,
pyrimidinyl, pyridazinyl, or pyridiyl (said phenyl, pyrazinyl,
pyrimidinyl, pyridazinyl, or pyridyl are optionally substituted
with halogen, hydroxy, cyano, amino, nitro, alkyl, alkoxy,
alkoxycarbonyl, alkylcarbonyl, or aminocarbonyl).
[0074] Item24. The compound according to anyone of Items 1-10, or
pharmaceutically acceptable salt thereof,
R.sub.3 and R.sub.4 are hydrogen.
[0075] Item25. The compound according to anyone of Items 1-24, or
pharmaceutically salt thereof,
R.sub.5 is
[0076] 1: hydrogen,
[0077] 2: halogen,
[0078] 3: cyano,
[0079] 4: nitro,
[0080] 5: hydroxy,
[0081] 6: alkyl,
[0082] 7: alkoxy,
[0083] 8: amino,
[0084] 9: alkylcarbonyl,
[0085] 10: alkoxycarbonyl,
[0086] 11: aminocarbonyl, and
[0087] 12: the following group:
##STR00014##
(wherein Y, R.sub.9a, R.sub.9b, R.sub.9c, and m are same as the
above definition), wherein said alkyl, alkoxy, amino,
alkylcarbonyl, alkoxycarbonyl, and aminocarbonyl are optionally
substituted with one or more substituent(s) selected from the group
consisting of
[0088] (a) alkyl,
[0089] (b) alkenyl,
[0090] (c) alkynyl,
[0091] (d) hydroxy,
[0092] (e) amino,
[0093] (f) nitro,
[0094] (g) cyano,
[0095] (h) halogen,
[0096] (i) alkoxy,
[0097] (j) alkylcarbonyl,
[0098] (k) alkoxycarbonyl,
[0099] (l) aminocarbonyl,
[0100] (m) aryl,
[0101] (n) heteroaryl,
[0102] (o) cycloalkyl, and
[0103] (p) heteroalicyclic).
[0104] Item26. The compound according to Item 25, or
pharmaceutically salt thereof,
R.sub.5 is hydrogen, or the following group:
##STR00015##
(wherein Y, R.sub.9a, R.sub.9b, R.sub.9c, and m are same as the
above definition).
[0105] Item27. The compound according to anyone of Items 1-26, or
pharmaceutically acceptable salt thereof,
Y is pyrrole, furan, thiophene, imidazole, pyrazole, oxazole,
isoxazole, thiazole, isothiazole, triazole, thiadiazole,
oxadiazole, pyrrolidine, piperidine, azepane, tetrahydrofuran,
oxane, or oxepane.
[0106] Item28. The compound according to Item 27, or
pharmaceutically acceptable salt thereof,
Y is pyrrole, furan, thiophene, imidazole, pyrazole, oxazole,
isoxazole, thiazole, isothiazole, triazole, thiadiazole, or
oxadiazole.
[0107] Item29. The compound according to Item 28, or
pharmaceutically acceptable salt thereof,
Y is furan, thiophene, imidazole, pyrazole, oxazole, thiazole,
triazole, thiadiazole, oxadiazole,
[0108] Item30. The compound according to Item 27, or
pharmaceutically acceptable salt thereof,
Y is piperidine.
[0109] Item31. The compound according to anyone of Items 1-30, or
pharmaceutically acceptable salt thereof,
R.sub.9a is piperidinyl, pyranyl, phenyl, thiophenyl, or
pyridyl(said pheny and pyridyl are optionally substituted with
hydroxy, amino, nitro, cyano, alkyl, alkoxyl, trifluoromethyl, and
halogen).
[0110] Item32. The compound according to anyone of Items 1-31, or
pharmaceutically acceptable salt thereof,
R.sub.9b and R.sub.9c are each dependently hydrogen, methyl, ethyl,
halogen, cyano, amino, nitro, hydroxy, or trifluoromethyl.
[0111] Item33. The compound according to Item 32, or
pharmaceutically acceptable salt thereof,
R.sub.9b is each dependently hydrogen, methyl, ethyl, halogen,
cyano, amino, nitro, hydroxy, or trifluoromethyl.
[0112] Item34. The compound according to anyone of Items 1-33, or
pharmaceutically acceptable salt thereof,
m is the integra 0.
[0113] Item35. The compound according to anyone of Items 1-24, or
pharmaceutically acceptable salt thereof,
R.sub.5 is selected from the group consisting of the following
group:
##STR00016##
R.sub.9a is piperidinyl, pyranyl, phenyl, thiophenyl, pyrazinyl,
pyrimidinyl, pyridazinyl, or pyridyl (said piperidinyl, pyranyl,
phenyl, thiophenyl, pyrazinyl, pyrimidinyl, pyridazinyl, or pyridyl
are optionally substituted with halogen, hydroxy, cyano, amino,
nitro, alkyl, alkoxy, alkoxycarbonyl, alkylcarbonyl, or
aminocarbonyl), and R.sub.9b is alkyl.
[0114] Item36. The compound according to anyone of Items 1-24, or
pharmaceutically acceptable salt thereof,
R.sub.5 is selected from the group consisting of the following
group:
##STR00017##
and R.sub.9a is 4-piperazinyl (said piperidinyl is substituted with
alkyl, or alkylcarbonyl), pyrazinyl, pyrimidinyl, pyridazinyl, or
pyridiyl (said phenyl, pyrazinyl, pyrimidinyl, pyridazinyl, or
pyridyl are optionally substituted with halogen, hydroxy, cyano,
amino, nitro, alkyl, alkoxy, alkoxycarbonyl, alkylcarbonyl, or
aminocarbonyl).
[0115] Item37. The compound according to anyone of Items 1-35, or
pharmaceutically acceptable salt thereof,
R.sub.6 is hydrogen, halogen, methyl, ethyl, n-propyl, isopropyl,
trifluoromethyl, optionally substituted alkoxy, optionally
substituted amino, optionally substituted alkoxycarbonyl,
optionally substituted alkylcarbonyl, or optionally substituted
aminocarbonyl.
[0116] Item38. The compound according to anyone of Items 1-36, or
pharmaceutically acceptable salt thereof,
R.sub.7 is hydrogen, methyl, ethyl, n-propyl, or isopropyl.
[0117] Item39. The compound according to anyone of Items 1-10,
19-21, 36 or 37, or pharmaceutically acceptable salt thereof,
R.sub.3 is the following group:
##STR00018##
(wherein X, R.sub.8a, R.sub.8b, R.sub.8c, and n are same as the
above definition), R.sub.4 is hydrogen, and R.sub.5 is
hydrogen.
[0118] Item40. The compound according to anyone of Items 1-10,
19-21, 36 or 37, or pharmaceutically acceptable salt thereof,
R.sub.3 is hydrogen, R.sub.4 is the following group:
##STR00019##
(wherein X, R.sub.8a, R.sub.8b, R.sub.8c, and n are same as the
above definition), and R.sub.5 is hydrogen.
[0119] Item41. The compound according to anyone of Items 1-10,
31-34, 36 or 37, or pharmaceutically acceptable salt thereof,
R.sub.3 is hydrogen, R.sub.4 is hydrogen, and R.sub.5 is the
following group:
##STR00020##
(Y, R.sub.9a, R.sub.9b, R.sub.9c, and m are same as the above
definition).
[0120] Item42. The compound according to Item1, or pharmaceutically
acceptable salt thereof, wherein a compound of formula (1) is
compounds of the following formulae:
##STR00021##
(wherein R.sub.7, Y, R.sub.9a, R.sub.9b, R.sub.9c, m, and R.sub.10
are same as the above definition).
[0121] Item43. The compound according to Item1, or pharmaceutically
acceptable salt thereof, wherein a compound of formula (1) is
compounds of the following formulae:
##STR00022##
(wherein R.sub.7, R.sub.9a, and R.sub.10 are same as the above
definition).
[0122] Item44. The compound according to Item1, or pharmaceutically
acceptable salt thereof, wherein a compound of formula (1) is
compounds of the following formulae:
##STR00023##
(wherein R.sub.7, X, R.sub.8a, R.sub.8b, R.sub.8c, n, and R.sub.10
are same as the above definition).
[0123] Item45. The compound according to Item1, or pharmaceutically
acceptable salt thereof, wherein a compound of formula (1) is
compounds of the following formulae:
##STR00024##
(wherein R.sub.7, X, R.sub.8a, R.sub.8b, R.sub.8c, n, and R.sub.10
are same as the above definition).
[0124] Item46. The compound according to Item1, or pharmaceutically
acceptable salt thereof, selected from the group consisting of:
##STR00025## ##STR00026## ##STR00027##
[0125] Item47. A pharmaceutical composition comprising the compound
anyone of Items 1-46, or pharmaceutically acceptable salt
thereof.
[0126] Item48. The pharmaceutical composition according to Item 47,
for treating cancer in a mammal, comprising to the mammal in need
thereof a therapeutically effective amount of said pharmaceutical
composition.
[0127] Item49. The pharmaceutical composition according to Item 48,
wherein the cancer is selected from the group consisting of lung
cancer, bladder cancer, head and neck cancer, melanoma, ovarian
cancer, prostate cancer, breast cancer, small-cell lung cancer,
glioma, colorectal cancer, non-small cell lung cancer,
genitourinary cancer, pancreatic cancer, thyroid cancer, Hodgkin's
lymphoma, non-Hodgkin's lymphoma, gastrointestinal cancer, gastric
cancer, hepatoma, gastrointestinal stromal tumor, squamous cell
carcinoma, renal cell carcinoma, astrocytoma, Kaposi's sarcoma,
chronic myelogenous leukemia, acute myelogenous leukemia,
myeloproliferative disorders, and glioblastoma.
[0128] Item50. A method for treating cancer in a mammal, comprising
administering to the mammal in need thereof a therapeutically
effective amount of the active ingredient the compound of any one
of Items 1-46, or pharmaceutically acceptable salt thereof.
[0129] Item51. A method for treating cancer in a mammal according
to Item 50, wherein the cancer is selected from the group
consisting of lung cancer, bladder cancer, head and neck cancer,
melanoma, ovarian cancer, prostate cancer, breast cancer,
small-cell lung cancer, glioma, colorectal cancer, non-small cell
lung cancer, genitourinary cancer, pancreatic cancer, thyroid
cancer, Hodgkin's lymphoma, non-Hodgkin's lymphoma,
gastrointestinal cancer, gastric cancer, hepatoma, gastrointestinal
stromal tumor, squamous cell carcinoma, renal cell carcinoma,
astrocytoma, Kaposi's sarcoma, chronic myelogenous leukemia, acute
myelogenous leukemia, myeloproliferative disorders, and
glioblastoma.
Effect of the Invention
[0130] A compound of formula (1), or a pharmaceutically acceptable
salt thereof is useful a CSCPKs inhibitor to inhibit, reduce or
diminish cancer stem cell survival and/or proliferation in a
mammal. A compound of formula (1), or a pharmaceutically acceptable
salt thereof is also useful an anti cancer agent.
DESCRIPTION OF EMBODIMENT
[0131] The present invention is described in more detail below. The
term "group" as used in the specification refers to a monovalent
group. For example, the term "alkyl group" refers to a monovalent
saturated hydrocarbon group. Moreover, the term "group" is
sometimes omitted in the explanation of substituents in the
specification.
[0132] The number of substituents in groups defined by the phrase
"optionally substituted" or "substituted" is not particularly
limited as long as the substituents are replaceable, and the number
is one or more than one. Moreover, unless otherwise particularly
specified, the explanation of each group is also applicable when
the groups are part or substituents of other groups. Furthermore,
in the specification, groups that are not modified by the phrase
"optionally substituted" or "substituted" refer to "unsubstituted"
groups.
[0133] The term "alkyl" refers to a straight or branched chain
alkane (hydrocarbon) radical containing from 1 to 12 carbon atoms,
preferably 1 to 6 carbon atoms, more preferably 1 to 4 carbon
atoms. The example of "alkyl" groups include methyl, ethyl, propyl,
isopropyl, n-butyl, t-butyl, isobutyl pentyl, hexyl, isohexyl,
heptyl, 4, 4-dimethylpentyl, octyl, 2,2,4-trimethylpentyl, nonyl,
decyl, undecyl, dodecyl, and the like. "Optionally substituted
alkyl" refers to an alkyl group optionally substituted with one or
more substituents, preferably 1 to 4 substituents, at any available
point of attachment.
[0134] The substituents include, but are not limited to, one or
more of the following groups:
hydrogen, halogen, cyano, nitro, amino, cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, heteroalicyclic, aryl, heteroaryl, alkoxy,
thioxo, sulfoxide, alkoxysulfonyl, aminosulfonyl, alkoxycarbonyl
optionally substituted with hydroxyl or amino, alkylcarbonyl
optionally substituted amino, or aminocarbonyl optionally
substituted with hydroxyl or amino.
[0135] Preferable substituents are selected from the group
consisting of alkyl, alkenyl, alkynyl, hydroxy, amino, nitro,
cyano, halogen, alkoxy, alkylcarbonyl, alkoxycarbonyl,
aminocarbonyl, aryl, heteroaryl, cycloalkyl, and
heteroalicyclic.
[0136] The term "alkenyl" refers to a straight or branched chain
hydrocarbon radical containing from 2 to 12 carbon atoms and at
least one carbon-carbon double bond. The such groups include, for
example, ethenyl or allyl.
[0137] The term "alkynyl" refers to a straight or branched chain
hydrocarbon radical containing from 2 to 12 carbon atoms and at
least one carbon to carbon triple bond. Examples of such group
include, for example, ethinyl.
[0138] The term "cycloalkyl" refers to a fully saturated cyclic
hydrocarbon group containing from 1 to 4 rings and 3 to 8 carbons
per ring. Examples of such group include, for example, cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl, or cycloheptyl.
[0139] "Optionally substituted cycloalkyl" refers to a cycloalkyl
group substituted with one or more substituents, preferably 1 to 4
substituents, at any available point of attachment. The examples of
substituent in optionally substituted cycloalkyl include, but are
not limited to, nitro, cyano, or alkyl optionally substituted with
hydroxy or alkoxy.
[0140] The "alkyl" moiety of "alkoxy" has the same meaning as
defined in the "alkyl".
[0141] Preferable one is a straight or branched chain alkoxy
radical containing from 1 to 12 carbon atoms, preferably 1 to 6
carbon atoms, more preferably 1 to 4 carbon atoms. Examples of
"alkoxy" include methoxy, ethoxy, propoxy, isopropoxy, butoxy,
isobutoxy, sec-butoxy, or tert-butoxy.
[0142] "Optionally substituted alkoxy" refers to an alkoxy group
optionally substituted with one or more substituents, preferably 1
to 4 substituents, at any available point of attachment.
[0143] The substituents include, but are not limited to, one or
more of the following groups:
hydrogen, halogen, cyano, nitro, amino, cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, heteroalicyclic, aryl, heteroaryl, alkoxy,
thioxo, sulfoxide, alkoxysulfonyl, aminosulfonyl, alkoxycarbonyl
optionally substituted with hydroxyl or amino, alkylcarbonyl
optionally substituted amino, or aminocarbonyl optionally
substituted with hydroxyl or amino.
[0144] Preferable substituents are selected form the groups
consisting of alkyl, alkenyl, alkynyl, hydroxy, amino, nitro,
cyano, halogen, alkoxy, alkylcarbonyl, alkoxycarbonyl,
aminocarbonyl, aryl, heteroaryl, cycloalkyl, and
heteroalicyclic
[0145] The term "aryl" refers to cyclic, aromatic hydrocarbon
groups that have 1 to 5 aromatic rings, especially monocyclic or
bicyclic groups such as phenyl, biphenyl or naphthyl.
[0146] Where containing two or more aromatic rings (bicyclic, etc),
the aromatic rings of the aryl group may be joined at a single
point (e.g., biphenyl), or fused (e.g., naphthyl, phenanthrenyl and
the like).
[0147] "Optionally substituted aryl" refers to an aryl group
substituted by one or more substituents, preferably 1 to 3
substituents, at any point of attachment.
[0148] Examples of substituent include, but are not limited to,
halogen, hydroxy, cyano, amino, nitro, optionally substituted with
alkyl, optionally substituted with alkenyl, optionally substituted
with alkynyl, optionally substituted with alkoxy, alkoxycarbonyl,
alkylcarbonyl, or aminocarbonyl.
[0149] Preferable substituents are selected from the groups
consisting of hydroxy, cyano, amino, nitro, alkyl, alkenyl,
alkynyl, alkoxy, alkoxycarbonyl, alkylcarbonyl, and
aminocarbonyl.
[0150] The term "heteroaryl" refers to 5 to 12-membered mono- or
poly-cyclic aromatic group, and comprises the same or different and
one or more (e.g., 1 to 4) heteroatoms selected from nitrogen atom,
sulfur atom or oxygen atom. A preferable "polycyclic heteroaryl" is
bi- or tri-cyclic group, more preferably bicyclic group. The
polycyclic heteroaryl includes a condensed ring of the above
monocyclic heteroaryl with an aromatic ring (including benzene,
pyridine) or a non-aromatic ring (including cyclohexyl). The term
"five membered heteroaryl" refers to 5-membered mono-cyclic
aromatic group, and comprises the same or different and one or more
(e.g., 1 to 4) heteroatoms selected from nitrogen atom, sulfur atom
or oxygen atom.
[0151] Concrete examples of the "heteroaryl" include, but not
limited to, pyrrole, furan, thiophene, imidazole, pyrazole,
oxazole, isoxazole, thiazole, isothiazole, triazole, thiadiazole,
oxadiazole, pyrrolidine, piperidine, azepane, tetrahydrofuran,
oxane, or oxepane.
[0152] "Optionally substituted heteroaryl" and "optionally
substituted five membered heteroaryl" refer to an heteroaryl group
substituted by one or more substituents, preferably 1 to 3
substituents, at any point of attachment, more preferably 1 or
2.
[0153] Examples of substituent include, but are not limited to,
halogen, hydroxy, cyano, amino, nitro, optionally substituted with
alkyl, optionally substituted with alkenyl, optionally substituted
with alkynyl, optionally substituted with alkoxy, alkoxycarbonyl,
alkylcarbonyl, or aminocarbonyl.
[0154] Preferable substituents are selected from the groups
consisting of hydroxy, cyano, amino, nitro, alkyl, alkenyl,
alkynyl, alkoxy, alkoxycarbonyl, alkylcarbonyl, and
aminocarbonyl.
[0155] The term "heteroalicyclic" refers to fully saturated, or
partially or fully unsaturated, including aromatic (i.e.,
"heteroaryl") cyclic groups (for example, 4 to 7 membered
monocyclic, 7 to 11 membered bicyclic, or 8 to 16 membered
tricyclic ring systems) which have at least one heteroatom in at
least one carbon atom-containing ring. Each ring of the
heterocyclic group containing a heteroatom may have 1, 2, 3, or 4
heteroatoms selected from nitrogen atoms, oxygen atoms and/or
sulfur atoms, where the nitrogen and sulfur heteroatoms may
optionally be oxidized and the nitrogen heteroatoms may optionally
be quaternized. (The term "heteroarylium" refers to a heteroaryl
group bearing a quaternary nitrogen atom and thus a positive
charge.) The heterocyclic group may be attached to the remainder of
the molecule at any heteroatom or carbon atom of the ring or ring
system.
[0156] Examples of monocyclic heteroalicyclic group include
azetidinyl, pyrrolidinyl, pyrrolyl, pyrazolyl, oxetanyl,
pyrazolinyl, imidazolyl, imidazolinyl, imidazolidinyl, oxazolyl,
oxazolidinyl, isoxazolinyl, isoxazolyl, thiazolyl, thiadiazolyl,
thiazolidinyl, isothiazolyl, isothiazolidinyl, furyl,
tetrahydrofuryl, thienyl, oxadiazolyl, piperidinyl, piperazinyl,
2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolodinyl,
2-oxoazepinyl, azepinyl, hexahydrodiazepinyl, 4-piperidonyl,
pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, triazolyl,
tetrazolyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl,
thiomorpholinyl sulfoxide, thiomorpholinyl sulfone, 1,3-dioxolane
or tetrahydro-1,1-dioxothienyl.
[0157] Examples of bicyclic heteroalicyclic group include indolyl,
isoindolyl, benzothiazolyl, benzoxazolyl, benzoxadiazolyl,
benzothienyl, benzo[d][1,3]dioxolyl,
2,3-dihydrobenzo[b][1,4]dioxinyl, quinuclidinyl, quinolinyl,
tetrahydroisoquinolinyl, isoquinolinyl, benzimidazolyl,
benzopyranyl, indolizinyl, benzofuryl, benzofurazanyl, chromonyl,
coumarinyl, benzopyranyl, cinnolinyl, quinoxalinyl, indazolyl,
pyrrolopyridyl, furopyridinyl (such as furo[2,3-c]pyridinyl,
furo[3,2-b]pyridinyl] or furo[2,3-b]pyridinyl), dihydroisoindolyl,
dihydroquinazolinyl (such as 3,4-dihydro-4-oxo-quinazolinyl),
triazinylazepinyl, or tetrahydroquinolinyl. Exemplary tricyclic
heterocyclic groups include carbazolyl, benzidolyl,
phenanthrolinyl, acridinyl, phenantliridinyl, or xanthenyl.
[0158] "Optionally substituted heteroalicyclic" refers to
heteroalicyclic groups substituted with one or more substituents,
preferably 1 to 4 substituents, at any available point of
attachment.
[0159] Examples of substituent include, but are not limited to,
halogen, hydroxy, cyano, amino, nitro, optionally substituted with
alkyl, optionally substituted with alkenyl, optionally substituted
with alkynyl, optionally substituted with alkoxy, alkoxycarbonyl,
alkylcarbonyl, or aminocarbonyl.
[0160] Preferable substituents are selected from the groups
consisting of hydroxy, cyano, amino, nitro, alkyl, alkenyl,
alkynyl, alkoxy, alkoxycarbonyl, alkylcarbonyl, and amino
carbonyl.
[0161] The terms "halogen" or "halo" refer to chlorine, bromine,
fluorine or iodine.
[0162] The "alkoxycarbonyl" is a group wherein the "alkoxy" binds
to carbonyl.
[0163] Preferable "alkoxycarbonyl" is the group having 1 to 12
carbon atoms, preferably 1 to 6 carbon atoms, more preferably 1 to
4 carbon atoms.
[0164] Specifically, it includes methoxycarbonyl, ethoxycarbonyl,
propoxycarbonyl, 2-propoxycarbonyl, or tert-butoxycarbonyl.
[0165] "Optionally substituted alkoxycarbonyl" refers to an alkoxy
group optionally substituted with one or more substituents,
preferably 1 to 2 substituents, at any available point of
attachment.
[0166] The substituents include, but are not limited to, one or
more of the following groups:
hydrogen, halogen, cyano, nitro, amino, cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, heteroalicyclic, aryl, heteroaryl, alkoxy,
thioxo, sulfoxide, alkoxysulfonyl, aminosulfonyl, alkoxycarbonyl
optionally substituted with hydroxyl or amino, alkylcarbonyl
optionally substituted amino, or aminocarbonyl optionally
substituted with hydroxyl or amino.
[0167] Preferable substituents are selected form the groups
consisting of alkyl, alkenyl, alkynyl, hydroxy, amino, nitro,
cyano, halogen, alkoxy, alkylcarbonyl, alkoxycarbonyl,
aminocarbonyl, aryl, heteroaryl, cycloalkyl, and
heteroalicyclic
[0168] The "alkylcarbonyl" means a group wherein the "alkyl" binds
to carbonyl.
[0169] Preferable "alkylcarbonyl" is the group having 1 to 12
carbon atoms, preferably 1 to 6 carbon atoms, more preferably 1 to
4 carbon atoms.
[0170] Specifically, it includes acetyl, propionyl or butyryl.
[0171] "Optionally substituted alkylcarbonyl" refers to an alkoxy
group optionally substituted with one or more substituents,
preferably 1 to 2 substituents, at any available point of
attachment.
[0172] The substituents include, but are not limited to, one or
more of the following groups:
hydrogen, halogen, cyano, nitro, amino, cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, heteroalicyclic, aryl, heteroaryl, alkoxy,
thioxo, sulfoxide, alkoxysulfonyl, aminosulfonyl, alkoxycarbonyl
optionally substituted with hydroxyl or amino, alkylcarbonyl
optionally substituted amino, or aminocarbonyl optionally
substituted with hydroxyl or amino.
[0173] Preferable substituents are selected from the groups
consisting of alkyl, alkenyl, alkynyl, hydroxy, amino, nitro,
cyano, halogen, alkoxy, alkylcarbonyl, alkoxycarbonyl,
aminocarbonyl, aryl, heteroaryl, cycloalkyl, and
heteroalicyclic
[0174] The substituent of "optionally substituted amino" refers to
an amino group optionally substituted with one or more
substituents, preferably 1 or 2 substituent(s), at any available
point of attachment.
[0175] The substituents include, but are not limited to, one or
more of the following groups:
halogen, cyano, nitro, amino, cycloalkyl, alkenyl, cycloalkenyl,
alkynyl, heteroalicyclic, aryl, heteroaryl, alkoxy, thioxo,
sulfoxide, alkoxysulfonyl, aminosulfonyl, alkoxycarbonyl optionally
substituted with hydroxyl or amino, alkylcarbonyl optionally
substituted amino, or aminocarbonyl optionally substituted with
hydroxyl or amino.
[0176] Preferable substituents are selected form the groups
consisting of alkyl, alkenyl, alkynyl, hydroxy, amino, nitro,
cyano, halogen, alkoxy, alkylcarbonyl, alkoxycarbonyl,
aminocarbonyl, aryl, heteroaryl, cycloalkyl, and
heteroalicyclic.
[0177] The "optionally substituted amino" in the "optionally
substituted aminocarbonyl" has the same meaning as defined in the
"optionally substituted amino".
[0178] A preferable embodiment in the present invention is
explained in more detail.
[0179] R.sub.1 is preferably optionally substituted heteroaryl, or
optionally substituted heteroalicyclic,
[0180] wherein the heteroaryl is selected from the group consisting
of pyrrole, furan, thiophene, pyridine, pyrimidine, pyrazine,
pyridazine, indole, and pyrrolopyridinone, and
[0181] heteroalicyclic is selected from the group consisting of
pyridone, pyrrolidine, and piperidine, more preferably,
[0182] optionally substituted heteroaryl, or optionally substituted
heteroalicyclic,
[0183] wherein heteroaryl is pyrrole, or pyrrolopyridinone, and
[0184] heteroalicyclic is pyridine.
[0185] R.sub.1 is more preferably one of the group selected from
the following group:
##STR00028##
(wherein R.sub.10 is same as the above definition).
[0186] R.sub.10 is preferably hydroxy, mono-substituted alkyl amino
(said alkyl is substituted by di-substituted alkyl amino, or
heteroalicyclic), piperazynyl, homopiperazynyl or morphorinyl, more
preferably,
[0187] hydroxy, ethyl amino (said ethyl is substituted by di-ethyl
amino, 1-pyrrolidino), or 4-piperazinyl (said piperazinyl is
substituted with alkyl), more preferably,
[0188] hydroxy, 2-(di-ethyl amino)ethyl amino, 2-pyrrolidino ethyl
amino, 4-methyl piperazinyl or morpholino.
[0189] R.sub.2 is preferably hydrogen, or optionally substituted
alkyl, more preferably, hydrogen.
[0190] R.sub.3, and R.sub.4 are preferably each independently
1: hydrogen, 2: halogen, 3: cyano, 4: nitro, 5: hydroxy, 6: alkyl.
7: alkoxy, 8: amino, 9: alkylcarbonyl, 10: alkoxycarbonyl, 11:
aminocarbonyl, and 12: the following group:
##STR00029##
(wherein X, R.sub.8a, R.sub.8b, R.sub.8c, and n are same as the
above definition), wherein said alkyl, alkoxy, amino,
alkylcarbonyl, alkoxycarbonyl, and aminocarbonyl are optionally
substituted with one or more substitutent(s) selected from the
group consisting of (a) alkyl, (b) alkenyl, (c) alkynyl, (d)
hydroxy, (e) amino, (f) nitro, (g) cyano, (h) halogen, (i) alkoxy,
(j) alkylcarbonyl, (k) alkoxycarbonyl, (l) aminocarbonyl, (m) aryl,
(n) heteroaryl, (o) cycloalkyl, and (p) heteroalicyclic),
[0191] more preferably, R.sub.3, and R.sub.4 are each independently
1: hydrogen, 2: halogen, 3: alkyl(said alkyl is optionally
substituted with one or more substituent(s) selected from the group
consisting of (a) alkyl, (b) alkenyl, (c) alkynyl, (d) hydroxy, (e)
amino, (f) nitro, (g) cyano, (h) halogen, (i) alkoxy, (j)
alkylcarbonyl, (k) alkoxycarbonyl, (l) aminocarbonyl, (m) aryl, (n)
heteroaryl, (l) cycloalkyl, and (o) heteroalicyclic), or 4: the
following group:
##STR00030##
(wherein X, R.sub.8a, R.sub.8b, R.sub.8c, and n are same as the
above definition), more preferably, R.sub.3, and R.sub.4 are each
independently hydrogen, or the following group:
##STR00031##
(wherein X, R.sub.8a, R.sub.8b, R.sub.8c, and n are same as the
above definition).
[0192] X is preferably thiophene, pyrazole, oxazole, thiazole,
thiadiazole, or oxadiazole,
[0193] more preferably, X is pyrazole, thiazole, or oxazole.
[0194] X is preferably piperidine.
[0195] R.sub.8a, R.sub.8b and R.sub.8c are preferably each
independently hydrogen, alkyl, halogen, cyano, amino, nitro,
hydroxy, trifluoromethyl, phenyl, or pyridyl,
[0196] more preferably, R.sub.8a, R.sub.8b and R.sub.8c are each
dependently hydrogen, alkyl, phenyl, or pyridyl.
[0197] n is preferably the integra 0.
[0198] In case that X is selected from the group consisting of the
following group:
##STR00032##
[0199] R.sub.8a is preferably piperidinyl, pyranyl, phenyl,
thiophenyl, pyrazinyl, pyrimidinyl, pyridazinyl, or pyridyl (said
piperidinyl, pyranyl, phenyl, thiophenyl pyrazinyl, pyrimidinyl,
pyridazinyl, or pyridyl are optionally substituted with halogen,
hydroxy, cyano, amino, nitro, alkyl, alkoxy, alkoxycarbonyl,
alkylcarbonyl, or aminocarbonyl), and
[0200] R.sub.8b is preferably alkyl.
[0201] In case that X is selected from the group consisting of the
following group:
##STR00033##
and
[0202] R.sub.8a is preferably 4-piperazinyl (said piperidinyl is
substituted with alkyl or alkylcarbonyl), 4-tetrahydropyranyl,
phenyl, pyrazinyl, pyrimidinyl, pyridazinyl, or pyridiyl (said
phenyl, pyrazinyl, pyrimidinyl, pyridazinyl, or pyridyl are
optionally substituted with halogen, hydroxy, cyano, amino, nitro,
alkyl, alkoxy, alkoxycarbonyl, alkylcarbonyl, or
aminocarbonyl).
[0203] In a compound of formula (1), R.sub.5 is 1: hydrogen, 2:
halogen, 3: cyano, 4: nitro, 5: hydroxy, 6: alkyl, 7: alkoxy, 8:
amino, 9: alkylcarbonyl, 10: alkoxycarbonyl, 11: aminocarbonyl, and
12: the following group:
##STR00034##
(wherein Y, R.sub.9a, R.sub.9b, R.sub.9c, and m are same as the
above definition),
[0204] wherein said alkyl, alkoxy, amino, alkylcarbonyl,
alkoxycarbonyl, and aminocarbonyl are optionally substituted with
one or more substituent(s) selected from the group consisting of
(a) alkyl, (b) alkenyl, (c) alkynyl, (d) hydroxy, (e) amino, (f)
nitro, (g) cyano, (h) halogen, (i) alkoxy, (j) alkylcarbonyl, (k)
alkoxycarbonyl, (l) aminocarbonyl, (m) aryl, (n) heteroaryl, (l)
cycloalkyl, and (o) heteroalicyclic, more preferably,
[0205] R.sub.5 is hydrogen, or the following group:
##STR00035##
(wherein Y, R.sub.9a, R.sub.9b, R.sub.9c, and m are same as the
above definition). Y is preferably pyrrole, furan, thiophene,
imidazole, pyrazole, oxazole, isoxazole, thiazole, isothiazole,
triazole, thiadiazole, or oxadiazole,
[0206] more preferably, Y is furan, thiophene, imidazole, pyrazole,
oxazole, thiazole, triazole, thiadiazole, oxadiazole.
[0207] Y is preferably piperidine.
[0208] R.sub.9a is preferably phenyl or pyridyl(said pheny and
pyridyl are optionally substituted with hydroxy, amino, nitro,
cyano, alkyl, alkoxyl, trifluoromethyl, and halogen.
[0209] R.sub.9b and R.sub.9c are preferably each dependently
hydrogen, methyl, ethyl, halogen, cyano, amino, nitro, hydroxy, or
trifluoromethyl.
[0210] R.sub.9b is preferably each dependently hydrogen, methyl,
ethyl, halogen, cyano, amino, nitro, hydroxy, or
trifluoromethyl.
[0211] m is preferably the integra 0.
[0212] In case that R.sub.5 is selected from the group consisting
of the following group:
##STR00036##
[0213] R.sub.9a is preferably piperidinyl, pyranyl, phenyl,
thiophenyl, pyrazinyl, pyrimidinyl, pyridazinyl or pyridyl (said
piperidinyl, pyranyl, phenyl, thiophenyl pyrazinyl, pyrimidinyl,
pyridazinyl, or pyridyl are optionally substituted with halogen,
hydroxy, cyano, amino, nitro, alkyl, alkoxy, alkoxycarbonyl,
alkylcarbonyl, or aminocarbonyl), and
[0214] R.sub.9b is preferably alkyl.
[0215] In case that Y is selected from the group consisting of the
following group:
##STR00037##
and
[0216] R.sub.9a is 4-piperazinyl (said piperidinyl is substituted
with alkyl or alkylcarbonyl), 4-tetrahydropyranyl, phenyl,
pyrazinyl, pyrimidinyl, pyridazinyl, or pyridiyl (said phenyl,
pyrazinyl, pyrimidinyl, pyridazinyl, or pyridyl are optionally
substituted with halogen, hydroxy, cyano, amino, nitro, alkyl,
alkoxy, alkoxycarbonyl, alkylcarbonyl, or aminocarbonyl).
[0217] R.sub.6 is preferably hydrogen halogen cyano, or optionally
substituted alkyl,
[0218] more preferably, R.sub.6 is hydrogen.
[0219] R.sub.7 is hydrogen, methyl, ethyl, n-propyl, or
isopropyl,
[0220] more preferably, R.sub.7 is hydrogen.
[0221] Preferable compound in the present invention is the compound
selected the group consisting of the following:
##STR00038## ##STR00039## ##STR00040##
[0222] In case of compound of formula (1), preferably,
[0223] R.sub.3 and R.sub.4 are each hydrogen, and
[0224] R.sub.5 is the following group:
##STR00041##
(wherein Y, R.sub.9a, R.sub.9b, R.sub.9c, and m are same as the
above definition); or
[0225] R.sub.3 is the following group:
##STR00042##
(wherein X, R.sub.8a, R.sub.8b, R.sub.8c, and n are same as the
above definition), and
[0226] R.sub.4 and R.sub.5 are each hydrogen; or
[0227] R.sub.3 is hydrogen,
[0228] R.sub.4 is the following group:
##STR00043##
(wherein X, R.sub.8a, R.sub.8b, R.sub.8c, and n are same as the
above definition), and
[0229] R.sub.5 is hydrogen,
[0230] A compound of formula (1) is preferably compounds explained
in the following (1) to (4). Each definition in these compounds is
the same as defined above. A preferable embodiment of the
definition is also the same as defined above.
(1) A compound of the following formula (1-a), or a
pharmaceutically acceptable salt thereof.
##STR00044##
(2) A compound of the following formula (1-b), or a
pharmaceutically acceptable salt thereof.
##STR00045##
(3) A compound of the following formula (1-c), or a
pharmaceutically acceptable salt thereof.
##STR00046##
(4) A compound of the following formula (1-d), or a
pharmaceutically acceptable salt thereof.
##STR00047##
(5) A compound of the following formula (1-e), or a
pharmaceutically acceptable salt thereof.
##STR00048##
(6) A compound of the following formula (1-f), or a
pharmaceutically acceptable salt thereof.
##STR00049##
(7) A compound of the following formula (1-g), or a
pharmaceutically acceptable salt thereof.
##STR00050##
(8) A compound of the following formula (1-h), or a
pharmaceutically acceptable salt thereof.
##STR00051##
[0231] Preparation methods for a compound of formula (1) are
explained. A compound formula (1) or a pharmaceutically acceptable
salt thereof is illustrated, but the present invention is not
intended to be limited thereto.
[0232] In the following method, the starting materials and the
intermediates of the reaction may be isolated and purified if
desired using conventional techniques, including but not limited to
filtration, distillation, crystallization, chromatography and the
like.
[0233] The materials of invention can be characterized by using
conventional means including but not limited to physical constants
and spectral data. The reactions are performed in solvents
appropriate to the reagents and materials employed and are suitable
for transformations being effected. The representative examples
include, but are not limited to, tetrahyrdofuran, dimethylforamide,
methanol, ethanol, water, dimethylforamide, chloroform,
dichloromethane, hexane, toluene, 1,4-dioxane or ethyl acetate.
[0234] Unless specified, the reactions described herein were
performed at atmospheric pressure over a temperature range from
about -78.degree. C. to about 150.degree. C.
[0235] For heating, any methods can be used which depends on
reagent and target material. The representative examples include,
but are not limited to, water bath, oil bath, water bath, or
microwave reactor.
[0236] The compound of formula (1) in the present invention may be
prepared from known compounds by optionally combining the method of
the following Preparation methods I to II, similar methods to the
following Preparation methods, or synthetic known to a skilled
person.
Preparation of Method
[0237] A compound of formula (1) may be synthesized by the
following method.
##STR00052##
[In the scheme, R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5,
R.sub.6 and R.sub.7 are as defined in the above item 1. A is
optionally substituted heteroaryl group in R.sub.3, R.sub.4 and
R.sub.5 in the above item 1. X is metal containing group such as
boronic acid, boronic acid pinacol ester, trifluoro boran, organic
tin, zinc halide, magnesium halide, organic silicon, and organic
lithium. Y is leaving group such as Cl, Br, I, and OTf.]
Preparation of Method I
[0238] A compound of formula (1) may be synthesized by the
following method.
[0239] Among a compound of formula (1), Compound 1-3 or a
pharmaceutically acceptable salt thereof is prepared by the
following method.
##STR00053##
[In the scheme, R.sub.1, R.sub.2, and R.sub.7 are as defined in the
above item 1. A is optionally substituted heteroaryl group in
R.sub.3, R.sub.4 and R.sub.5 in the above item 1. X is metal
containing group such as boronic acid, boronic acid pinacol ester,
trifluoro boran, organic tin, zinc halide, magnesium halide,
organic silicon, and organic lithium. Y is leaving group such as
Cl, Br, I, and OTf.]
[0240] A compound of formula 1-1 can react with a compound of
formula 1-2 in the presence of transition metal catalyst
(representative examples include, but are not limited to
tetrakis(triphenylphosphine)palladium(0),
[1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride,
palladium carbon, dichlorobis(triphenylphosphine)nickel(II), or
bis(triphenylphosphine)palladium(II) dichloride.), alkali metal
carbonate (representative examples include, but are not limited to
potassium carbonate, sodium carbonate, or cesium carbonate.) or
other alkali metal salt (sodium hydroxide, potassium hydroxide,
dodium ethoxide, sodium methoxide, sodium tert-butoxide, potassium
tert-butoxide, sodium hydride, sodium phosphate, potassium
phosphate.) and appropriate solvent or without solvent to give a
compound of formula 1-3.
Preparation Method II
[0241] A compound I-1 may be prepared from a compound II-2.
##STR00054##
[In the scheme, the symbols have the same meaning as defined
above.]
[0242] A compound of formula II-1 can react with a compound of
formula II-2 in the presence of a base (representative examples
include, but are not limited to pyrrolidine and piperidine) or an
acid (representative examples include, but are not limited to
hydrochloric acid, acetic acid, trifluoroacetic acid), and
appropriate solvent or without solvent to give a compound of
formula I-1.
Preparation Method III
[0243] A compound I-3 may be prepared from a compound III-1.
##STR00055##
[In the scheme, the symbols have the same meaning as defined
above.]
[0244] A compound of formula III-1 can react with a compound of
formula III-2 in the presence of transition metal catalyst
(representative examples include, but are not limited to,
tetrakis(triphenylphosphine)palladium(0),
[1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride,
palladium carbon, dichlorobis(triphenylphosphine)nickel(II), or
bis(triphenylphosphine)palladium(II) dichloride.), alkali metal
carbonate (representative examples include, but are not limited to,
potassium carbonate, sodium carbonate, or cesium carbonate.) or
other alkali metal salt (sodium hydroxide, potassium hydroxide,
dodium ethoxide, sodium methoxide, sodium tert-butoxide, potassium
tert-butoxide, sodium hydride, sodium phosphate, potassium
phosphate.), and appropriate solvent or without solvent to give a
compound of formula 1-3.
Preparation Method IV
[0245] A compound III-1 may be prepared from a compound IV-1.
##STR00056##
[In the scheme, the symbols have the same meaning as defined
above.]
[0246] A compound of formula IV-1 can react with a compound of
formula II-2 in the presence of a base (representative examples
include, but are not limited to pyrrolidine and piperidine) or an
acid (representative examples include, but are not limited to
hydrochloric acid, acetic acid, trifluoroacetic acid), and
appropriate solvent or without solvent to give a compound of
formula III-1.
Preparation Method V
[0247] A compound of formula III-1 may be prepared from a compound
I-1.
##STR00057##
[In the scheme, the symbols have the same meaning as defined
above.]
[0248] A compound of formula I-1 can react with a compound of boron
reagent (representative examples include, but are not limited to,
bis(pinacolato)diboron, bis(neopentyl Glycolato)diboron, or
bis(catecholato)diboron.) in the presence of transition metal
catalyst (representative examples include, but are not limited to,
dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium,
[1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride, or
bis(triphenylphosphine)palladium(II) dichloride.), alkali metal
carbonate or alkali metal acetate (representative examples include,
but are not limited to, potassium carbonate, sodium carbonate,
cesium carbonate, or potassium acetate.), and appropriate solvent
or without solvent to give a compound of formula III-1.
Preparation Method VI
[0249] A compound of formula I-3 may be prepared from a compound
VI-1.
##STR00058##
[In the scheme, the symbols have the same meaning as defined
above.]
[0250] A compound of formula VI-1 can react with a compound of
formula II-2 in the presence of a base (representative examples
include, but are not limited to pyrrolidine and piperidine) or an
acid (representative examples include, but are not limited to
hydrochloric acid, acetic acid, trifluoroacetic acid), and
appropriate solvent or without solvent to give a compound of
formula 1-3.
Preparation Method VII
[0251] A compound of formula VI-1 may be prepared from a compound
of formula II-1.
##STR00059##
[In the scheme, the symbols have the same meaning as defined
above.]
[0252] A compound of formula II-1 can react with a compound of
formula I-2 in the presence of transition metal catalyst
(representative examples include, but are not limited to,
tetrakis(triphenylphosphine)palladium(0),
[1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride,
palladium carbon, dichlorobis(triphenylphosphine)nickel(II), or
bis(triphenylphosphine)palladium(II) dichloride.), alkali metal
carbonate (representative examples include, but are not limited to,
potassium carbonate, sodium carbonate, or cesium carbonate.) or
other alkali metal salt (sodium hydroxide, potassium hydroxide,
dodium ethoxide, sodium methoxide, sodium tert-butoxide, potassium
tert-butoxide, sodium hydride, sodium phosphate, potassium
phosphate.), and appropriate solvent or without solvent to give a
compound of formula VI-1.
Preparation Method VIII
[0253] A compound of formula VI-1 may be prepared from a compound
of formula IV-1.
##STR00060##
[In the scheme, the symbols have the same meaning as defined
above.]
[0254] A compound of formula IV-1 can react with a compound of
formula III-2 in the presence of transition metal catalyst
(representative examples include, but are not limited to,
tetrakis(triphenylphosphine)palladium(0),
[1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride,
palladium carbon, dichlorobis(triphenylphosphine)nickel(II), or
bis(triphenylphosphine)palladium(II) dichloride), alkali metal
carbonate (representative examples include, but are not limited to,
potassium carbonate, sodium carbonate, or cesium carbonate) or
other alkali metal salt (sodium hydroxide, potassium hydroxide,
dodium ethoxide, sodium methoxide, sodium tert-butoxide, potassium
tert-butoxide, sodium hydride, sodium phosphate, potassium
phosphate), and appropriate solvent or without solvent to give a
compound of formula VI-1.
Preparation Method IX
[0255] A compound of formula IX-3 may be prepared from a compound
of formula IX-1.
##STR00061##
[In the scheme, Ra and Rb are each independently halogen, cyano,
nitro, amino, cycloalkyl, alkenyl, cycloalkenyl, alkynyl,
heteroalicyclic, aryl, heteroaryl, alkoxy, thioxo, sulfoxide,
alkoxysulfonyl, aminosulfonyl, alkoxycarbonyl optionally
substituted with hydroxyl or amino, alkylcarbonyl optionally
substituted amino, or aminocarbonyl optionally substituted with
hydroxyl or amino, B is optionally substituted heteroaryl group or
optionally substituted heteroalicyclic group.]
[0256] A compound of formula IX-1 can react with a compound of
formula IX-2 (representative examples include, but are not limited
to, N.sup.1,N.sup.1-diethylethane-1,2-diamine,
N.sup.1,N.sup.1-dimethylethane-1,2-diamine,
2-(pyrrolidin-1-yl)ethanamine, N-methyl-piperazine,
N-methyl-homopiperazine, 2-morpholinoethanamine, or morpholine.) in
the presence of coupling reagent (representative examples include,
but are not limited to, N,N-dicyclohexylcarbodiimide,
N,N-diisopropylcarbodiimide, or 1-ethyl-3-(3-dimethylaminopropyl)
carbodiimide hydrochloride.), and appropriate solvent or without
solvent to give a compound of formula IX-3. This amide formation
reaction can be performed in the presence of appropriate additives
(representative examples include, but are not limited to,
1-hydroxybenzotriazole, or N-hydroxysuccinimide).
Preparation Method X
[0257] A compound of formula X-5 and X-6 may be prepared from a
compound of formula IV-1.
##STR00062##
[In the scheme, R.sub.2, X and Y are same as the above definition.
C is optionally substituted heteroalicyclic group (said
heteroalicyclic group is unsaturated, and one of double bond is
attached to X or Y). D is optionally substituted heteroalicyclic
group (wherein heteroalicyclic group is saturated.). E is
optionally substituted hetero alicyclic. F is optionally
substituted heteroaryl or optionally substituted hetero alicyclic
group.]
[0258] A compound of formula IV-1 can react with a compound of
formula X-1 in the presence of transition metal catalyst
(representative examples include, but are not limited to,
tetrakis(triphenylphosphine)palladium(0),
[1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride,
palladium carbon, dichlorobis(triphenylphosphine)nickel(II), or
bis(triphenylphosphine)palladium(II) dichloride.), alkali metal
carbonate (representative examples include, but are not limited to,
potassium carbonate, sodium carbonate, or cesium carbonate) or
other alkali metal salt (sodium hydroxide, potassium hydroxide,
dodium ethoxide, sodium methoxide, sodium tert-butoxide, potassium
tert-butoxide, sodium hydride, sodium phosphate, potassium
phosphate), and appropriate solvent or without solvent to give a
compound of formula X-2.
[0259] A compound of formula X-2 can further react in the presence
of transition metal catalyst (representative examples include, but
are not limited to, palladium carbon, platinum carbon or rhodium
carbon.), and appropriate solvent or without solvent under hydrogen
atmosphere to give a compound of formula X-3. The reaction can be
performed in any hydrogen pressure which depends on reagent and
target material. However, preferable pressure is between 1 to 10
atm, and even more preferably between 1 to 5 atm.
[0260] A compound of formula X-3 can react with a compound of
formula X-4 in the presence of reducing reagent (representative
examples include, but are not limited to, sodium
triacetoxyborohydride, tetramethyl triacetoxyborohydride, picolyl
borane, or sodium cyanoborohydride.), acid (representative examples
include, but are not limited to acetic acid, or trifluoroacetic
acid), and appropriate solvent or without solvent to give a
compound of formula X-5.
[0261] A compound of formula X-3 can react with a compound of
formula X-6 (wherein Z is leaving group representative examples
include, but are not limited to, chloro, bromo, iodo,
trifluoromethanesulfonyl, or p-tosyl) in the presence of tertiary
amine (representative examples include, but are not limited to,
diisopyropylethylamine, triethylamine, or pyridine), and
appropriate solvent or without solvent to give a compound of
formula X-7.
Preparation Method XI
[0262] A compound of formula X-2 may be prepared from a compound of
formula II-2.
##STR00063##
[In the scheme, the symbols have the same meaning as defined
above.]
[0263] A compound of formula II-1 can react with a compound of
formula XI-1 in the presence of transition metal catalyst
(representative examples include, but are not limited to,
tetrakis(triphenylphosphine)palladium(0),
[1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride,
palladium carbon, dichlorobis(triphenylphosphine)nickel(II) or
bis(triphenylphosphine)palladium(II) dichloride.), alkali metal
carbonate (potassium carbonate, sodium carbonate, or cesium
carbonate) or other alkali metal salt (sodium hydroxide, potassium
hydroxide, dodium ethoxide, sodium methoxide, sodium tert-butoxide,
potassium tert-butoxide, sodium hydride, sodium phosphate,
potassium phosphate), and appropriate solvent or without solvent to
give a compound of formula X-2.
Preparation Method XII
[0264] A compound of formula XII-4 may be prepared from a compound
of formula XII-1.
##STR00064##
[In the scheme, R.sub.2 is same as the above definition. G is
defined as R.sub.8a or R.sub.9a.]
[0265] A compound of formula XII-1 can react with a compound of
formula XII-2 in the presence of coupling reagent (representative
examples include, but are not limited to,
N,N-dicyclohexylcarbodiimide, N,N-diisopropylcarbodiimide, or
1-ethyl-3-(3-dimethylaminopropyl.), primary or secondary amine
(representative examples include, but are not limited to,
2-amino-1-phenylethanone, 2-amino-1-p-tolylethanone,
2-amino-1-(4-chlorophenyl)ethanone,
2-amino-1-(4-methoxyphenyl)ethanone, or
2-amino-1-(pyridin-4-yl)ethanone.), and appropriate solvent or
without solvent to give a compound of formula XII-3. This amide
formation reaction can be performed in the presence of appropriate
additives (representative examples include, but are not limited to,
1-hydroxybenzotriazole, or N-hydroxysuccinimide.).
[0266] A compound of formula XII-3 can further react in the
presence of acid (representative examples include, but are not
limited to, trifluoroacetic acid, methanesulfonic acid,
p-toluenesulfonic acid, benzenesulfonic acid, or sulfuric acid) to
give compound of formula XII-4.
Preparation Method XIII
[0267] A compound of formula XIII-6 may be prepared from a compound
of formula XIII-1.
##STR00065##
[In the scheme, the symbols have the same meaning as defined
above.]
[0268] A compound of formula XIII-1 can react with azide salt
(representative examples include, but are not limited to, sodium
azide.), and appropriate solvent or without solvent to give a
compound of formula XIII-2. This reaction can be preformed in the
presence of additive (representative examples include, but are not
limited to, potassium iodide, or tetrabutylammonium iodide).
[0269] A compound of formula XIII-2 can further react in the
presence of metal catalyst (representative examples include, but
are not limited to, palladium carbon, or platinum carbon.), and
appropriate solvent or without solvent under hydrogen atmosphere to
give a compound of formula XIII-3. The reaction can be performed in
any hydrogen pressure which depends on reagent and target material.
However, preferable pressure is between 1 to 10 atm, and even more
preferably between 1 to 5 atm.
[0270] A compound of formula XIII-3 can further react with a
compound of formula XIII-4 (wherein "Z" is defined as leaving group
such as Cl, Br and the likes. Representative examples include, but
are not limited to, benzoyl chloride, benzoyl bromide,
4-chlorobenzoyl chloride, 4-methoxybenzoyl chloride,
4-methylbenzoyl chloride, isonicotinoyl chloride, nicotinoyl
chloride, picolinoyl chloride, or tetrahydro-2H-pyran-4-carbonyl
chloride.), and appropriate solvent or without solvent to give a
compound of formula XIII-5. This reaction can be preformed in the
presence of additive (representative examples include, but are not
limited to, diisopropylethylamine, pyridine, or
triethylamine.).
[0271] A compound of formula XIII-5 can further react in the
presence of acids (representative examples include, but are not
limited to, trifluoroacetic acid, methanesulfonic acid,
p-toluenesulfonic acid, benzenesulfonic acid, or sulfuric acid.),
and appropriate solvent or without solvent to give a compound of
formula XIII-6.
Preparation Method XIV
[0272] A compound of formula XIV-4 may be prepared from a compound
of formula XIV-1.
##STR00066##
[In the scheme, R is alkyl. The symbols have the same meaning as
defined above.]
[0273] A compound of formula XIV-1 can react with hydrazine
(representative examples include, but are not limited to, hydrazine
hydrate, or hydrazine) in the presence of solvent or without
solvent to give a compound of formula XIV-2.
[0274] A compound of formula XIV-2 can react with aryl nitrile
(representative examples include, but are not limited to,
benzonitrile, 4-methylbenzonitrile, 4-chlorobenzonitrile,
4-methoxybenzonitrile, 3-methylbenzonitrile, isonicotinonitrile, or
tetrahydro-2H-pyran-4-carbonitrile.) in the presence of alkali
metal carbonate (representative examples include, but are not
limited to, potassium carbonate, sodium carbonate, or cesium
carbonate.) and appropriate solvent or without solvent to give a
compound of formula XIV-4.
Preparation Method XV
[0275] A compound of formula XV-2 may be prepared from a compound
of formula XIII-1.
##STR00067##
[In the scheme, the symbols have the same meaning as defined
above.]
[0276] A compound of formula XIII-1 can react with a compound of
formula XV-1 (representative examples include, but are not limited
to, benzimidamide, substituted benzimidamide, or
isonicotinimidamide.), and appropriate solvent or without solvent
to give a compound of formula XV-2. This reaction can be preformed
in the presence of additive (representative examples include, but
are not limited to, sodium iodide or potassium iodide.).
Preparation Method XVI
[0277] A compound of formula XVI-2 may be prepared from a compound
of formula XIII-1.
##STR00068##
[In the scheme, the symbols have the same meaning as defined
above.]
[0278] A compound of formula XIII-1 can react with a compound of
formula XVI-1 (representative examples include, but are not limited
to, benzothioamide, 4-methylbenzothioamide, 4-chlorobenzothioamide,
4-methoxybenzothioamide, 3-methylbenzothioamide,
pyridine-4-carbothioamide, pyridine-3-carbothioamide,
pyridine-2-carbothioamide or tert-butyl
4-carbamothioylpiperidine-1-carboxylate), and appropriate solvent
or without solvent to give a compound of formula XVI-2.
Preparation Method XVII
[0279] A compound of formula XVII-3 may be prepared from a compound
of formula IV-1.
##STR00069##
[In the scheme, the symbols have the same meaning as defined
above.]
[0280] A compound of formula IV-1 can react with azide salt
(representative examples include, but are not limited to, sodium
azide, or hydrogen azide.) and a compound of formula XVII-2
(representative examples include, but are not limited to, phenyl
acetylene, 1-ethynyl-4-methylbenzene, 4-chloro-1-ethynyl-benzene,
or 4-ethynylpyridine.) in the presence of alkali base carbonate
(representative examples include, but are not limited to, sodium
carbonate, potassium carbonate, or cesium carbonate.), copper salt
(representative examples include, but are not limited to, copper
chloride (I), copper bromide (I), or copper iodide (I).), ascorbate
(representative examples include, but are not limited to, sodium
ascorbate, or potassium ascorbate.), amine (representative examples
include, but are not limited to, N,N-dimethylethylenediamine) and
appropriate solvent or without solvent to give a compound of
formula XVII-3.
sodium ascorbate
Preparation Method XVIII
[0281] A compound of formula XVIII-4 and XVIII-6 may be prepared
from a compound of formula IV-1.
##STR00070##
[In the scheme, the symbols have the same meaning as defined
above.]
[0282] A compound of formula IV-1 (wherein R is alkyl, or trialkyl
silyl) can react with a compound of formula XVIII-1 (representative
examples include, but are not limited to, phenylacetylene,
prop-1-yne, or 3,3-diethoxyprop-1-yne.) in the presence of
transition metal catalyst (representative examples include, but are
not limited to, tetrakis(triphenylphosphine)palladium(0),
[1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride, or
bis(triphenylphosphine)palladium(II) dichloride.), copper catalyst
(representative examples include, but are not limited to, copper
chloride (I), copper bromide (I), or copper iodide (I).), organic
base (representative examples include, but are not limited to,
diisopropylethyamine, or triethylamine.), and appropriate solvent
or without solvent to give a compound of formula XVIII-2.
[0283] A compound of formula XVIII-2 can further react with a
compound of formula XVIII-3 (representative examples include, but
are not limited to, phenylazide, 1-azido-4-methylbenzene,
1-azido-4-chlorobenzene, or 4-azidopyridine.) in the presence of
copper catalyst (representative examples include, but are not
limited to, copper chloride (I), copper bromide (I), or copper
iodide (I).), alkali metal carbonate (representative examples
include, but are not limited to, sodium carbonate, potassium
carbonate, or cesium carbonate.), amine (representative examples
include, but are not limited to, N,N-dimethylethylenediamine.) and
appropriate solvent or without solvent to give a compound of
formula XVIII-4.
[0284] A compound of formula XVIII-2 (wherein R contains ketone,
aldehyde or their equivalent (representative examples include, but
are not limited to, 5-(3,3-diethoxyprop-1-ynyl)indolin-2-one, or
5-(3,3-diethoxybut-1-ynyl)indolin-2-one.) next to triple bond) can
react with a compound of formula XVIII-5 (representative examples
include, but are not limited to, phenylhydrazine, p-tolylhydrazine,
or p-cyanophenylhydrazine.) in the presence of appropriate solvent
or without solvent to give a compound of formula XVIII-6. This
reaction can be performed in the presence of acid (representative
examples include, but are not limited to, sulfuric acid,
p-toluenesulfonyl acid, or methanesulfonyl acid.).
[0285] The present invention encompasses a compound of formula (1)
or a prodrug thereof, or a pharmaceutically acceptable salt
thereof. It also encompasses a solvate thereof such as a hydrate or
an ethanolate, etc. Further, the present invention encompasses
every tautomer, every existing stereoisomer and every crystalline
form of the compound of the present invention (1).
[0286] The term "prodrug of a compound of formula (1)" herein means
a compound which is converted to a compound of formula (1) by
reaction(s) by enzyme or gastric acid, etc. under the physiological
condition in vivo, e.g. a compound which is converted to a compound
of formula (1) by enzymatic oxidization, reduction, hydrolysis,
etc.; a compound which is converted to a compound of formula (1) by
hydrolysis by gastric acid, etc.
[0287] The compounds of the present invention which contain a basic
moiety, such as but not limited to an amine or a pyridine or
imidazole ring, may form salts with a variety of organic and
inorganic acids. Exemplary acid addition salts include acetates
(such as those formed with acetic acid or trihaloacetic acid, for
example, trifluoroacetic acid), adipates, alginates, ascorbates,
aspartates, benzoates, benzenesulfonates, bisulfates, borates,
butyrates, citrates, camphorates, camphorsulfonates,
cyclopentanepropionates, digluconates, dodecylsulfates, ethane
sulfonates, fumarates, glucoheptonoates, glycerophosphates,
hemisulfates, heptanoates, hexanoates, hydrochlorides,
hydrobromides, hydroiodides, hydroxyethanesulfonates (e.g.,
2-hydroxyethanesulfonates), lactates, maleates, methanesulfonates,
naphthalenesulfonates (e.g., 2-naphthalenesulfonates), nicotinates,
nitrates, oxalates, pectinates, persulfates, phenylpropionates
(e.g., 3-phenylpropionates), phosphates, picrates, pivalates,
propionates, salicylates, succinates, sulfates (such as those
formed with sulfuric acid), sulfonates, tartrates, thiocyanates,
toluenesulfonates such as tosylates, undecanoates, and the
like.
[0288] The compounds of the present invention which contain an
acidic moiety, such as but not limited to a carboxylic acid, may
form salts with a variety of organic and inorganic bases. Exemplary
basic salts include ammonium salts, alkali metal salts such as
sodium, lithium and potassium salts, alkaline earth metal salts
such as calcium and magnesium salts, salts with organic bases (for
example, organic amines) such as benzathines, dicyclohexylamines,
hydrabamines (formed with N, N-bis(dehydroabietyl)
ethylenediamine), N-methyl-D-glucamines, N-methyl-D-glycamides,
t-butyl amines, and salts with amino acids such as arginine, lysine
and the like. Basic nitrogen-containing groups may be quaternized
with agents such as lower alkyl halides {e.g. methyl, ethyl,
propyl, and butyl chlorides, bromides and iodides), dialkyl
sulfates (e.g. dimethyl, diethyl, dibutyl, and diamyl sulfates),
long chain halides (e.g. decyl, lauryl, myristyl and stearyl
chlorides, bromides and iodides), aralkyl halides (e.g. benzyl and
phenethyl bromides), and others.
[0289] Solvates of the compounds of the invention are also
contemplated herein. Solvates of the compounds of the present
invention include, for example, hydrates.
[0290] Compounds of the present invention, and salts thereof, may
exist in their tautomeric form (for example, as an amide or imino
ether). All such tautomeric forms are contemplated herein as part
of the present invention.
[0291] The compounds of the present invention may form salts which
are also within the scope of this invention. Reference to a
compound of the present invention herein is understood to include
reference to salts thereof, unless otherwise indicated. The term
"salt(s)", as employed herein, denotes acidic and/or basic salts
formed with inorganic and/or organic acids and bases. In addition,
when a compound of the present invention contains both a basic
moiety, such as but not limited to a pyridine or imidazole, and an
acidic moiety such as but not limited to a carboxylic acid,
zwitterions ("inner salts") may be formed and are included within
the term "salt(s)" as used herein. "Pharmaceutically acceptable
salts" (i.e., non-toxic, physiologically acceptable) are preferred,
although other salts are also useful, e.g., in isolation or
purification steps which may be employed during preparation. Salts
of the compounds of the present invention may be formed, for
example, by reacting a compound I, II or III with an amount of acid
or base, such as an equivalent amount, in a medium such as one in
which the salt precipitates or in an aqueous medium followed by
lyophilization.
[0292] All stereoisomers of the compounds of the present invention
(for example, those which may exist due to asymmetric carbons on
various substituents), including enantiomeric forms and
diastereomeric forms, are contemplated within the scope of this
invention. Individual stereoisomers of the compounds of the
invention may, for example, be substantially free of other isomers
(e.g., as a pure or substantially pure optical isomer having a
specified activity), or may be admixed, for example, as racemates
or with all other, or other selected, stereoisomers. The chiral
centers of the present invention may have the S or R configuration
as defined by the IUPAC 1974 Recommendations. The racemic forms can
be resolved by physical methods, such as, for example, fractional
crystallization, separation or crystallization of diastereomeric
derivatives or separation by chiral column chromatography. The
individual optical isomers can be obtained from the racemates by
any suitable method, including without limitation, conventional
methods, such as, for example, salt formation with an optically
active acid followed by crystallization.
[0293] Compounds of the present invention are, subsequent to their
preparation, preferably isolated and purified to obtain a
composition containing an amount by weight equal to or greater than
95% ("substantially pure" compound I), which is then used or
formulated as described herein. In certain embodiments, the
compounds of the present invention are more than 99% pure.
[0294] All configurational isomers of the compounds of the present
invention are contemplated, either in admixture or in pure or
substantially pure form. The definition of compounds of the present
invention embraces both cis (2) and trans (E) alkene isomers, as
well as cis and trans isomers of cyclic hydrocarbon or heterocyclic
rings.
[0295] Throughout the specifications, groups and substituents
thereof may be chosen to provide stable moieties and compounds.
[0296] The present invention also provides, in part, a method of
treating, preventing or ameliorating a protein kinase related
disorder in a mammal, comprising administering to the mammal in
need thereof a therapeutically effective amount of a pharmaceutical
composition comprising a compound of the present invention as
described hereinabove.
[0297] The mammal may be in need of the treatment or the treatment
may be administered prophylactically for prevention or for
amelioration of the protein kinase related disorder.
[0298] A "protein kinase related disorder" is any disease or
deleterious condition in which a protein kinase plays a role.
Examples include a serine-threonine kinase related disorder, a
receptor tyrosine kinase related disorder, a non-receptor tyrosine
kinase related disorder, an EGFR related disorder, an IGFR related
disorder, a PDGFR related disorder and a flk related disorder. The
compounds of the present invention may be used for any of these
protein kinase related disorders.
[0299] In certain embodiments, the protein kinase related disorder
is a cancer such as lung cancer, bladder cancer, head and neck
cancer, melanoma, ovarian cancer, prostate cancer, breast cancer,
small-cell lung cancer, glioma, colorectal cancer, non-small cell
lung cancer, genitourinary cancer, pancreatic cancer, thyroid
cancer, Hodgkin's lymphoma, non-Hodgkin's lymphoma,
gastrointestinal cancer, gastric cancer, hepatoma, gastrointestinal
stromal tumor, squamous cell carcinoma, renal cell carcinoma,
astrocytoma, Kaposi's sarcoma, chronic myelogenous leukemia, acute
myelogenous leukemia, myeloproliferative disorders, and
glioblastoma.
[0300] According to one or more embodiments of the present
invention, "cancer stem cell" ("CSC") or "cancer stem cells"
("CSCs") refer to a minute population of cancer cells that have
self-renewal capability and are tumorigenic. They are also called
"Cancer Initiating Cells", "Tumor Initiating Cells", "Cancer
Stem-Like Cells", "Stem-Like Cancer Cells", "aggressive cancer
cells", and "super malignant cancer cells", etc. The methods of
isolating these cells include but not limited to enrichment by
their ability of efflux Hoechst 33342, enrichment of surface
markers such as CD133, CD44, and others, and enrichment by their
tumorigenic property.
[0301] The term "CSCPK" or "CSCPKs" refer to protein kinase(s) that
are essential for cancer stem cell survival or self-renewal.
[0302] In certain embodiments, the protein kinase is CSCPK. The
compounds of the present invention are particularly useful for the
treatment, prevention or amelioration of cancer, such as lung
cancer, bladder cancer, head and neck cancer, melanoma, ovarian
cancer, prostate cancer, breast cancer, small-cell lung cancer,
glioma, colorectal cancer, non-small-cell lung cancer,
genitourinary cancer, pancreatic cancer, thyroid cancer, Hodgkin's
lymphoma, non-Hodgkin's lymphoma, gastrointestinal cancer, gastric
cancer, hepatoma, gastrointestinal stromal tumor, squamous cell
carcinoma, renal cell carcinoma, astrocytoma, Kaposi's sarcoma,
chronic myelogenous leukemia, acute myelogenous leukemia,
myeloproliferative disorders, and glioblastoma, by inhibiting
CSCPKs.
[0303] In yet other embodiments, the protein kinase includes
serine-threonine kinases, receptor tyrosine kinases and
non-receptor tyrosine kinases.
[0304] In yet other embodiments, the protein kinase related
disorder includes diabetes, an autoimmune disorder, a
hyperproliferation disorder, angiogenesis, an inflammatory
disorder, an immunological disorder, a cardiovascular disorder,
restenosis, fibrosis, psoriasis, von Heppel-Lindau disease,
osteoarthritis, neurodegeneration, infection, and rheumatoid
arthritis.
[0305] The present invention provides, in part, a method of
inhibiting/reducing/diminishing cancer stem cell survival and/or
proliferation, self-renewal in a mammal by inhibiting or decreasing
unwanted activity of several kinases including CSCPK.
[0306] The present invention also provides, in part, a method of
inhibiting cancer stem cell niche, or stromal cell signaling by
targeting CSCPKs.
[0307] The present invention further provides, in part, a method of
treating cancer, inhibiting/reducing/diminishing cancer stem cell
survival and/or proliferation.
[0308] The present invention also provides, in part, a method of
modulating the catalytic activity of a protein kinase. The method
comprises contacting said protein kinase with a compound of the
present invention, or a pharmaceutically-acceptable salt thereof.
In certain embodiments, the protein kinase includes a
serine-threonine kinase, a receptor tyrosine kinase and a
non-receptor tyrosine kinase.
[0309] The present invention also provides, in part, a
pharmaceutical composition comprising a compound of the present
invention as described hereinabove, or an enantiomer, diastereomer,
tautomer, or pharmaceutically acceptable salt, solvate or prodrugs
thereof, and a pharmaceutically-acceptable excipient, carrier, or
diluent.
[0310] Formulations of the present invention include those suitable
for oral, nasal, topical (including buccal and sublingual), rectal,
vaginal and/or parenteral administration. The formulations may
conveniently be presented in unit dosage form and may be prepared
by any methods well known in the art of pharmacy. The amount of
active ingredient which can be combined with a carrier material to
produce a single dosage form will vary depending upon the mammal
being treated and the particular mode of administration. The amount
of active ingredient, which can be combined with a carrier material
to produce a single dosage form, will generally be that amount of
the compound which produces a therapeutic effect. Generally, out of
100%, this amount will range, for example, from about 0.1% to about
25% of active ingredient.
[1] Therapeutic compositions or formulations of the invention
suitable for oral administration may be in the form of capsules,
cachets, pills, tablets, lozenges (using a flavored basis, usually
sucrose and acacia or tragacanth), powders, granules, or as a
solution or a suspension in an aqueous or non-aqueous liquid, or as
an oil-in-water or water-in-oil liquid emulsion, or as an elixir or
syrup, or as pastilles (using an inert base, such as gelatin and
glycerin, or sucrose and acacia) and/or as mouth washes and the
like, each containing a predetermined amount of a compound of the
present invention as an active ingredient. A compound of the
present invention may also be administered as a bolus, electuary or
paste.
[0311] In solid dosage forms of the invention for oral
administration (capsules, tablets, pills, dragees, powders,
granules and the like), the alcohol or inhibitor according to the
invention is mixed with one or more pharmaceutically-acceptable
carriers, such as sodium citrate or dicalcium phosphate, and/or any
of the following: fillers or extenders, such as starches, lactose,
sucrose, glucose, mannitol, and/or silicic acid; binders, such as,
for example, carboxymethylcellulose, alginates, gelatin, polyvinyl
pyrrolidone, sucrose and/or acacia; humectants, such as glycerol;
disintegrating agents, such as agar-agar, calcium carbonate, potato
or tapioca starch, alginic acid, certain silicates, sodium
carbonate, and sodium starch glycolate; solution retarding agents,
such as paraffin; absorption accelerators, such as quaternary
ammonium compounds; wetting agents, such as, for example, cetyl
alcohol, glycerol monostearate, and polyethylene
oxide-polypropylene oxide copolymer; absorbents, such as kaolin and
bentonite clay; lubricants, such a talc, calcium stearate,
magnesium stearate, solid polyethylene glycols, sodium lauryl
sulfate, and mixtures thereof; and coloring agents. In the case of
capsules, tablets and pills, the pharmaceutical compositions may
also comprise buffering agents. Solid compositions of a similar
type may also be employed as fillers in soft and hard-filled
gelatin capsules using such excipients as lactose or milk sugars,
as well as high molecular weight polyethylene glycols and the
like.
[0312] Liquid dosage forms for oral administration of the compounds
of the invention include pharmaceutically acceptable emulsions,
micro-emulsions, solutions, suspensions, syrups and elixirs. In
addition to the active ingredient, the liquid dosage forms may
contain inert diluents commonly used in the art, such as, for
example, water or other solvents, solubilizing agents and
emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl
carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate,
propylene glycol, 1,3-butylene glycol, oils (in particular,
cottonseed, groundnut, corn, germ, olive, castor and sesame oils),
glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty
acid esters of sorbitan, and mixtures thereof. Additionally,
cyclodextrins, for example, hydroxypropyl-beta-cyclodextrin, may be
used to solubilize compounds.
[0313] Besides inert diluents, the oral compositions can also
include adjuvants such as wetting agents, emulsifying and
suspending agents, sweetening, flavoring, coloring, perfuming and
preservative agents. Suspensions, in addition to the alcohols or
inhibitors according to the invention, may contain suspending
agents as, for example, ethoxylated isostearyl alcohols,
polyoxyethylene sorbitol and sorbitan esters, microcrystalline
cellulose, aluminum metahydroxide, bentonite, agar-agar and
tragacanth, and mixtures thereof.
[0314] Formulations of the pharmaceutical compositions of the
invention for rectal or vaginal administration may be presented as
a suppository, which may be prepared by mixing one or more alcohols
or inhibitors according to the invention, with one or more suitable
nonirritating excipients or carriers comprising, for example, cocoa
butter, polyethylene glycol, a suppository wax or a salicylate, and
which is solid at room temperature, but liquid at body temperature
and, therefore, will melt in the rectum or vaginal cavity and
release the active pharmaceutical agents of the invention.
Formulations of the present invention which are suitable for
vaginal administration also include pessaries, tampons, creams,
gels, pastes, foams or spray formulations containing such carriers
as are known in the art to be appropriate.
[0315] Dosage forms for the topical or transdermal administration
of an alcohol or other inhibitor according to the invention include
powders, sprays, ointments, pastes, creams, lotions, gels,
solutions, patches and inhalants. The active compound may be mixed
under sterile conditions with a pharmaceutically-acceptable
carrier, and with any preservatives, buffers, or propellants which
may be required.
[0316] The ointments, pastes, creams and gels may contain, in
addition to an alcohol or other inhibitor according to the
invention, excipients, such as animal and vegetable fats, oils,
waxes, paraffins, starch, tragacanth, cellulose derivatives,
polyethylene glycols, silicones, bentonites, silicic acid, talc and
zinc oxide, or mixtures thereof.
[0317] Powders and sprays can contain, in addition to a compound of
this invention, excipients such as lactose, talc, silicic acid,
aluminum hydroxide, calcium silicates and polyamide powder, or
mixtures of these substances. Sprays can additionally contain
customary propellants, such as chlorofluorohydrocarbons and
volatile unsubstituted hydrocarbons, such as butane and
propane.
[0318] Ophthalmic formulations, eye ointments, powders, solutions
and the like, are also contemplated as being within the scope of
this invention.
[0319] Pharmaceutical compositions of this invention suitable for
parenteral administration comprise one or more alcohols or
inhibitors according to the invention in combination with one or
more pharmaceutically-acceptable sterile isotonic aqueous or
nonaqueous solutions, dispersions, suspensions or emulsions, or
sterile powders which may be reconstituted into sterile injectable
solutions or dispersions just prior to use, which may contain
antioxidants, buffers, bacteriostats, solutes which render the
formulation isotonic with the blood of the intended recipient or
suspending or thickening agents.
[0320] In some cases, in order to prolong the effect of the alcohol
or inhibitor according to the invention, it is desirable to slow
the absorption of the alcohol or inhibitor from subcutaneous or
intramuscular injection. This may be accomplished by the use of a
liquid suspension of crystalline or amorphous material having poor
water solubility.
[0321] The rate of absorption of the drug then depends upon its
rate of dissolution, which, in turn, may depend upon crystal size
and crystalline form. Alternatively, delayed absorption of a
parenterally-administered composition is accomplished by dissolving
or suspending the alcohol or inhibitor in an oil vehicle. One
strategy for depot injections includes the use of polyethylene
oxide-polypropylene oxide copolymers wherein the vehicle is fluid
at room temperature and solidifies at body temperature.
[0322] The pharmaceutical compounds of this invention may be
administered alone, or simultaneously, subsequently or sequentially
with one or more active agents, other pharmaceutical agents, or
with other anti-cancer or cytotoxic agent as described hereinabove,
as well as in combination with a pharmaceutically-acceptable
excipient, carrier, or diluent as described above.
[0323] The amount of pharmacological agent in the oral unit dosage
form, with as a single or multiple dosage, is an amount that is
effective for treating a neurological disorder. As one of skill in
the art will recognize, the precise dose to be employed will depend
on a variety of factors, examples of which include the condition
itself, the seriousness of the condition being treated, the
particular composition used, as well as various physical factors
related to the individual being treated. In vitro or in vivo assays
can optionally be employed to help identify optimal dosage
ranges.
[0324] The compounds of the invention will normally be administered
in a daily dosage regimen (for an adult patient) of, for example,
an oral dose of between 1 mg and 2000 mg, preferably between 30 mg
and 1000 mg, e.g. between 10 and 250 mg or an intravenous,
subcutaneous, or intramuscular dose of between 0.1 mg and 100 mg,
preferably between 0.1 mg and 50 mg, e.g. between 1 and 25 mg of
the compounds of the invention or a pharmaceutically acceptable
salt thereof calculated as the free base, the compound being
administered 1 to 4 times per day. Suitably the compounds will be
administered for a period of continuous therapy, for example for a
week or more.
EXAMPLES
[0325] The following preparations and examples are given to enable
those skilled in the art to more clearly understand and to practice
the present invention. They should not be considered as limiting
the scope of the invention, but merely as being illustrative and
representative thereof.
[0326] The synthesized compounds are identified by NMR, LCMS, IR,
melting point, HPLC and so on.
[0327] The abbreviations used in NMR spectrums are following:
s for singlet, d for doublet, t for triplet, m for multiplet, br
for broad, brs for broad singlet, and J for binding constant.
Reference Example 1
Production of 5-(5-phenylthiophen-2-yl)indolin-2-one
##STR00071##
[0329] To a solution of 5-bromooxindole (100 mg, 0.572 mmol) in
dioxane/H.sub.2O (3 ml/1 ml) was added Pd(PPh.sub.3).sub.4 (55 mg,
0.047 mmol), 5-phenylthiophene-2-boronic acid (106 mg, 0.519 mmol)
and potassium carbonate (196 mg, 1.42 mmol). The mixture was
stirred at 120.degree. C. for 1 hour under microwave irradiation.
The residue was extracted with CHCl.sub.3, and the organic layer
was washed with H.sub.2O and brine, dried over Na.sub.2SO.sub.4 and
concentrated in vacuo. The residue was purified by column
chromatography (CHCl.sub.3/MeOH) to give
5-(5-phenylthiophen-2-yl)indolin-2-one (44 mg) as a pale yellow
solid.
[0330] MS m/z 292.4 (M+H).
Reference Examples 2 to 8
[0331] Reactions and treatments were carried out in the same manner
as Reference example 1 using the corresponding starting material
compounds, thereby givining the compounds of Reference example 2 to
8 shown in Table 1.
TABLE-US-00001 TABLE 1 Reference Example structure Spectral data 2
##STR00072## LCMS m/z 290.3 (M + H) 3 ##STR00073## 300 MHz
.sup.1H-NMR (DMSO-d.sub.6, .delta.) 10.48 (s, 1H), 8.12 (s, 1H),
8.03-7.98 (m, 2H), 7.65-7.49 (m, 5H), 7.29 (d, 1H, J = 7.7 Hz),
3.52 (s, 2H) 4 ##STR00074## 300 MHz .sup.1H-NMR (DMSO-d.sub.6,
.delta.) 9.97 (s, 1H), 8.20 (s, 1H), 8.06-8.00 (m, 2H), 7.75 (d,
1H, J = 7.5 Hz,), 7.61-7.50 (m, 3H), 7.25 (d, 1H, J = 7.2 Hz), 7.06
(dd, 1H, J = 7.2, 7.5 Hz), 3.60 (s, 2H) 5 ##STR00075## LCMS m/z
298.2 (M + H) 6 ##STR00076## LCMS m/z 292.4 (M + H) 7 ##STR00077##
LCMS m/z 291.3 (M + H) 8 ##STR00078## LCMS m/z 291.3 (M + H)
Reference Example 9
Production of 5-(5-phenyl-1,3,4-thiadiazol-2-yl)indolin-2-one
##STR00079##
[0333] To a solution of
5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indolin-2-one (98
mg, 0.38 mmol) in dioxane (0.76 ml) was added PdCl.sub.2(dppf)
CH.sub.2Cl.sub.2 (28 mg, 0.039 mmol),
2-bromo-5-phenyl-1,3,4-thiadiazole (138 mg, 0.57 mmol) and 2 M
potassium carbonate (aq, 568 .mu.L). The mixture was stirred at
90.degree. C. for 4 hour. The residue was extracted with EtOAc, and
the organic layer was washed with H.sub.2O and brine, dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The residue was
purified by column chromatography (n-hexane/EtOAc) to give
5-(5-phenyl-1,3,4-thiadiazol-2-yl)indolin-2-one (28 mg) as brown
oil.
[0334] LCMS m/z 294.3 (M+H)
Reference Examples 10 to 14
[0335] Reactions and treatments were carried out in the same manner
as Reference example 9 using the corresponding starting material
compounds, thereby givining the compounds of Reference example 10
to 12 shown in Table 2.
TABLE-US-00002 TABLE 2 Reference Example structure Spectral data 10
##STR00080## LCMS m/z 276.3 (M + H) 11 ##STR00081## LCMS m/z 293.2
(M + H) 12 ##STR00082## LCMS m/z 277.3 (M + H) 13 ##STR00083## 300
MHz .sup.1H-NMR (CDCl.sub.3, .delta.) 7.61 (s, 1H), 7.46 (s, 1H),
4.40-4.30 (m, 1H), 4.33-4.09 (m, 2H), 3.50 (d, 2H, J = 2.4, 8.8
Hz), 2.11-1.93 (m, 2H). 14 ##STR00084## 400 MHz .sup.1H-NMR
(CDCl.sub.3, .delta.) 8.58 (brs, 1H), 7.78 (d, 1H, J = 1.8 Hz),
7.65 (dd, 2H, J = 1.9, 6.8 Hz), 7.47 (dd, 2H, J = 1.9, 6.8 Hz),
7.16 (s, 1H), 7.09 (dd, 1H, J = 1.6, 8.1 Hz), 6.88 (d, 1H, J = 1.6,
8.1 Hz), 6.50 (d, 1H, J = 1.8 Hz), 3.56 (s, 2H).
Reference Example 15
Production of 5-(5-phenyloxazol-2-yl)indolin-2-one
##STR00085##
[0337] To a solution of 2-oxoindoline-5-carboxylic acid (1.3 g, 7.3
mmol) in DMF (50 ml) was added iPr.sub.2NEt (3.8 ml, 22 mmol), HOBt
(1.2 g, 8.8 mmol), WSCI (1.7 g, 8.8 mmol) and
2-amino-1-phenylethanone hydrochloride (1.3 g, 7.3 mmol). The
reaction mixture was stirred for 2 h at room temperature. The
mixture was poured into H.sub.2O and EtOAc. The resulting
precipitate was removed by filtration, and the filtrate was
separated. The organic layer was washed with sat. NaHCO.sub.3
solution, sat. NH.sub.4Cl solution and brine, and then dried over
Na.sub.2SO.sub.4. The solvent was evaporated and the residue (0.73
g) was used for the next reaction without further purification.
[0338] Sulfuric acid (5 ml) was added to the residue, and the
mixture was heated for 2 h at 100.degree. C. Ice was added, and the
mixture was extracted with EtOAc. The organic layer was washed with
H.sub.2O and brine, dried over Na.sub.2SO.sub.4 and evaporated. The
residue was crystallized from EtOH to afford
5-(5-phenyloxazol-2-yl)indolin-2-one (0.27 g, 13%).
[0339] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 10.68 (s, 1H),
7.96-7.91 (m, 2H), 7.84-7.80 (m, 2H), 7.77 (s, 1H), 7.52-7.47 (m,
2H), 7.37 (m, 1H), 6.97 (d, 1H, J=8.0 Hz), 3.60 (s, 2H).
Reference Example 16
Production of 5-(2-phenyloxazol-5-yl)indolin-2-one
##STR00086##
[0341] To a solution of 5-(2-chloroacetyl)indolin-2-one (1.0 g, 4.8
mmol) in DMF (20 ml) was added NaI (0.14 g, 0.96 mmol) and
NaN.sub.3 (0.37 g, 5.7 mmol), and the mixture was stirred for 2 h
at room temperature. H.sub.2O and EtOAc were added to the mixture,
and the resulting precipitate was filtered and dried to afford
5-(2-azidoacetyl)indolin-2-one (0.38 g, 37%).
[0342] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 10.77 (s, 1H),
7.84 (dd, 1H, J=8.2, 1.6 Hz), 7.79 (d, 1H, J=1.6 Hz), 6.93 (d, 1H,
J=8.2 Hz), 4.80 (s, 2H), 3.57 (s, 2H).
[0343] To a solution of 5-(2-azidoacetyl)indolin-2-one (0.20 g, 1.1
mmol) in DMF (5 ml) was added 10% Pd--C (0.20 g), and the mixture
was stirred for 3.5 h at room temperature under H.sub.2 atmosphere.
The mixture was passed through Celite. To the filtrate was added
benzoyl chloride (0.12 ml, 1.1 mmol) and iPr.sub.2NEt (0.36 ml, 2.2
mmol), and the reaction mixture was stirred for 1 h at 0.degree. C.
H.sub.2O and EtOAc were added to the mixture, and insoluble solid
was removed by filtration. The filtrate was separated and the
organic layer was washed with H.sub.2O and brine, dried over
Na.sub.2SO.sub.4 and evaporated. The residue was dissolved in
sulfuric acid (2.0 ml) and the mixture was heated for 2 h at
90.degree. C. The mixture was cooled to room temperature, and
H.sub.2O was added. The mixture was extracted with EtOAc, washed
with H.sub.2O and brine, dried over Na.sub.2SO.sub.4 and
evaporated. Purification by column chromatography (EtOAc/hex) gave
5-(2-phenyloxazol-5-yl)indolin-2-one (0.07 g, 24%).
[0344] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 10.58 (s, 1H),
8.08-8.05 (m, 2H), 7.72-7.65 (m, 3H), 7.58-7.50 (m, 3H), 6.92 (d,
1H, J=8.1 Hz), 3.57 (s, 2H).
Reference Example 17
Production of 5-(3-phenyl-1H-1,2,4-triazol-5-yl)indolin-2-one
##STR00087##
[0346] To a solution of methyl 2-oxoindoline-5-carboxylate (0.40 g,
4.8 mmol) in EtOH (8 ml) was added hydrazine monohydrate (2 ml),
and the mixture was stirred for 6 h at 80.degree. C. The mixture
was cooled to room temperature, and the resulting precipitate was
filtered and dried to afford 2-oxoindoline-5-carbohydrazide (0.25
g, 63%).
[0347] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.58 (s, 1H),
7.71-7.67 (m, 2H), 6.83 (d, 1H, J=8.0 Hz), 4.41 (br, 2H), 3.51 (s,
2H).
[0348] To a solution of 2-oxoindoline-5-carbohydrazide (200 mg,
1.05 mmol) in n-BuOH/DMF (6 ml/2 ml) was added benzonitrile (324
mg, 3.14 mmol) and potassium carbonate (29 mg, 0.21 mmol). The
mixture was heated at 150.degree. C. for 3 hours under microwave
irradiation. CHCl.sub.3/MeOH (20 ml/1 ml) was added to the mixture
and insoluble solid was removed by filtration. The filtrate was
concentrated. H.sub.2O was added to the residue and extracted with
CHCl.sub.3. The organic layer was dried over Na.sub.2SO.sub.4 and
concentrated in vacuo. Purification by column chromatography
(CHCl.sub.3/MeOH) gave
5-(3-phenyl-1H-1,2,4-triazol-5-yl)indolin-2-one (15 mg).
[0349] LCMS m/z 277.3 (M+H)
Reference Example 18
Production of 5-(2-phenyl-1H-imidazol-5-yl)indolin-2-one
##STR00088##
[0351] To a solution of 5-(2-chloroacetyl)indolin-2-one (100 mg,
0.477 mmol) in THF/H.sub.2O (3 ml/1 ml) was added benzimidamide
hydrochloride (75 mg, 0.477 mmol) and potassium carbonate (198 mg,
1.43 mmol). The mixture was stirred for 7 hours under reflux. The
mixture was extracted with CHCl.sub.3, and the organic layer washed
with H.sub.2O, dried over Na.sub.2SO.sub.4 and concentrated in
vacuo. The residue purified by column chromatography
(CHCl.sub.3/MeOH) to give
5-(3-phenyl-1H-imidazol-5-yl)indolin-2-one (19 mg).
[0352] LCMS m/z 276.30 (M+H)
Reference Example 19
Production of 5-(4-phenyl-1H-1,2,3-triazol-1-yl)indolin-2-one
##STR00089##
[0354] The mixture of 5-bromoindolin-2-one (530 mg, 2.5 mmol),
N,N-dimethylethylenediamine (44 mg, 0.5 mmol), ethynylbenzene (274
.mu.l, 2.5 mmol), CuI (48 mg, 0.25 mmol), sodium azide (325 mg, 5
mmol) and sodium ascorbate (99 mg, 0.5 mmol) in EtOH (7 ml),
H.sub.2O (3 ml) was heated to 80.degree. C. for 18 h. All reagents
were re-added and heated to 80.degree. C. for 10 h. After
confirming the reaction complete, reaction mixture was cooled to
room temperature and EtOH was removed under reduced pressure. 20 ml
of water was added and filtered. The filtrate was washed with water
and hexane and dried under vacuo to give
5-(4-phenyl-1H-1,2,3-triazol-1-yl)indolin-2-one (450 mg).
Reference Example 20
Production of 5-(1-phenyl-1H-1,2,3-triazol-4-yl)indolin-2-one
##STR00090##
[0356] To a solution of 5-iodoindolin-2-one (518 mg, 2 mmol), TEA
(3 ml) and CuI (38 mg) in DMF (3 ml) was added to
PdCl.sub.2(PPh.sub.3).sub.2 (70 mg). The mixture was cooled to
0.degree. C. and a solution of TMS-acetylene (1 ml). The mixture
was maintained same temperature for 3 h, then warmed to rt. After
stirring for overnight, the reaction mixture was concentrated in
vacuo. The residue was purified by silica gel column chromatography
to give 5-((trimethylsilyl)ethynyl)indolin-2-one (451 mg).
[0357] 7.37-7.34 (2H, m), 6.80 (1H, d, J=9.0 Hz), 3.51 (2H, s), and
0.24 (9H, s).
[0358] To a mixture of iodobenzene (204 mg, 1 mmol), sodium azide
(130 mg, 2 mmol), sodium carbonate (53 mg, 0.5 mmol), CuI (19 mg,
0.1 mmol), sodium ascorbate (20 mg) and
N,N'-dimethylethylenediamine (18 ul, 0.2 mmol) in EtOH (1.5 ml) and
water (0.5 ml) were added 5-((trimethylsilyl)ethynyl)indolin-2-one
(115 mg, 0.5 mmol), and stirred at 80.degree. C. for 2 h. After
cooling to ambient temp, EtOH was removed under reduced pressure.
The residue was suspended in EtOH and stirred for 1 h at rt and
filtered. The filtrate was washed with water and hexane and dried
under vacuo to give 5-(1-phenyl-1H-1,2,3-triazol-4-yl)indolin-2-one
(106 mg).
Reference Example 21
Production of 5-(1-phenyl-1H-pyrazol-5-yl)indolin-2-one
##STR00091##
[0360] To a solution of 5-iodo-2-oxoindoline (497 mg, 1.9 mmol) in
THF (20 ml) were added triethylamine (0.80 ml, 5.7 mmol),
3,3-diethoxyprop-1-yne (738 mg, 5.7 mmol), CuI (73 mg, 0.38 mmol)
and Pd(PPh.sub.3).sub.4 (222 mg, 0.19 mmol). The reaction mixture
was stirred for 4 h at 50.degree. C. The mixture was poured into
H.sub.2O and EtOAc. The mixture was separated into an aqueous layer
and an organic layer. The aqueous layer was extracted with ethyl
acetate 3 times. The combined organic layer was washed with sat.
NaHCO.sub.3 solution, and brine, and then dried over
Na.sub.2SO.sub.4. The solvent was evaporated and the residue
purified by column chromatography (EtOAc then CHCl.sub.3/MeOH) to
give 5-(3,3-diethoxyprop-1-ynyl)indolin-2-one as a brown solid (292
mg, 59%).
[0361] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.73 (s, 1H),
7.19 (d, 1H, J=8.0 Hz), 7.16 (s, 1H), 6.63 (d, 1H, J=8.0 Hz), 5.31
(s, 1H), 3.64 (dq, 2H, J=9.4, 7.1 Hz), 3.48 (dq, 2H, J=9.4, 7.1
Hz), 3.34 (s, 2H), 1.10 (t, 6H, J=7.1 Hz).
[0362] To a solution of 5-(3,3-diethoxyprop-1-ynyl)indolin-2-one
(100 mg, 0.39 mmol) in acetonitrile (5 ml) were added phenyl
hydrazine (38 .mu.L, 0.38 mmol) and sulfuric acid (52 .mu.L, 0.98
mmol), and the mixture was stirred for 3 h at room temperature,
then the mixture was stirred for 2 h at 50.degree. C. The reaction
mixture was poured into water (50 mL), and the resulting
precipitate was filtered and dried. The precipitated was dissolved
in acetonitrile (5 mL), then water (52 .mu.L, 3.9 mmol) and
sulfuric acid (93 .mu.L, 1.75 mmol) were added. The mixture was
heated at 80 C for 4 h. The mixture was cooled to room temperature,
and then neutralized with sat. NaHCO.sub.3. The mixture was
extracted with CHCl.sub.3/EtOAc 3 times. The combined organic
extracts were washed with sat. NaCl, dried over Na.sub.2SO.sub.4,
and evaporated in vacuo. The residue purified by column
chromatography (EtOAc/n-hexane) to give the title compound as a
brown solid (44 mg, 41%).
[0363] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.28 (brs, 1H),
7.73 (d, 1H, J=1.8 Hz), 7.39-7.28 (m, 5H), 7.13-7.08 (m, 2H), 6.81
(d, 1H, J=8.0 Hz), 6.48 (d, 1H, J=1.8 Hz), 3.51 (s, 2H).
[0364] MS m/z 276.3 (M+H)
Reference Example 22
Production of
5-(1-(tetrahydro-2H-pyran-4-yl)piperidin-4-yl)indolin-2-one
##STR00092##
[0366] To a suspension of 5-bromoindolin-2-one (600 mg, 2.83 mmol)
in 1,4-Dioxane (9 ml) and H.sub.2O (3 ml) were added tert-butyl
4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5,6-dihydropyridine-1(2H)-
-carboxylate (1.05 g, 3.40 mmol), Pd(PPh.sub.3).sub.4 (164 mg,
0.142 mmol) and K.sub.2CO.sub.3 (1.17 g, 8.50 mmol). After stirring
at 120.degree. C. in microwave reactor for 1 h, the reaction
mixture was diluted with sat. NaHCO.sub.3 aq. and extracted with
CHCl.sub.3. The organic layer was dried over Na.sub.2SO.sub.4 and
concentrated. The residue was purified by column chromatography
(CHCl.sub.3/MeOH) to give tert-butyl
4-(2-oxoindolin-5-yl)-5,6-dihydropyridine-1(2H-carboxylate (912 mg)
as mixture with triphenylphosphin oxide.
[0367] LCMS m/z 315 (M+H)
[0368] To a solution of tert-butyl
4-(2-oxoindolin-5-yl)-5,6-dihydropyridine-1(2H-carboxylate (912 mg,
2.90 mmol) in THF (10 ml) and MeOH (10 ml) was added 10% Pd/C (453
mg) and stirred at room temperature under H.sub.2 (1 atom)
atmosphere for 7 h. The reaction mixture was filtered through a
Celite pad and concentrated. The residue was purified by column
chromatography (CHCl.sub.3/MeOH) to afford tert-butyl
4-(2-oxoindolin-5-yl)piperidine-1-carboxylate (846 mg, 92%).
[0369] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 10.25 (s, 1H),
7.65-7.49 (m, 1H), 7.07 (s, 1H), 7.00 (d, 1H, J=7.8 Hz), 6.71 (d,
1H, J=7.8 Hz), 4.10-3.96 (m, 2H), 3.41 (s, 2H), 2.86-2.66 (m, 2H),
2.66-2.50 (m, 1H), 1.75-1.62 (m, 2H), 1.51-1.30 (m, 2H), 1.40 (s,
9H).
[0370] To a solution of TFA (10 ml) was added tert-butyl
4-(2-oxoindolin-5-yl)piperidine-1-carboxylate (789 mg, 2.49 mmol)
and stirred at room temperature for 30 min. The reaction mixture
was concentrated. The residue was diluted with 1N HCl and extracted
with CHCl.sub.3. The aqueous layer was added with 28% NH.sub.3 aq
until pH 8 and extracted with CHCl.sub.3/EtOH (3/1). The organic
layer was dried over Na.sub.2SO.sub.4 and concentrated to give
5-(piperidin-4-yl)indolin-2-one (409 mg, 76%).
[0371] LCMS m/z 217 (M+H)
[0372] To a solution of 5-(piperidin-4-yl)indolin-2-one (64.6 mg,
0.299 mmol) in THF (1.5 ml) and MeOH (3 ml) were added
dihydro-2H-pyran-4(3H)-one (0.132 ml, 1.34 mmol), acetic acid
(0.170 ml, 29.5 mmol) and NaBH.sub.3(CN) (61.6 mg, 0.931 mmol).
After stirring at room temperature for 4 days, the reaction mixture
was concentrated. The residue was diluted with sat. NaHCO.sub.3 aq.
and extracted with CHCl.sub.3. The organic layer was dried over
Na.sub.2SO.sub.4 and concentrated. The residue was purified by
column chromatography (CHCl.sub.3/MeOH) to give
5-(1-(tetrahydro-2H-pyran-4-yl)piperidin-4-yl)indolin-2-one (84.9
mg, 95%).
[0373] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 10.23 (s, 1H),
7.06 (s, 1H), 7.00 (d, 1H, J=7.9 Hz), 6.70 (d, 1H, J=7.9 Hz),
3.89-3.85 (m, 2H), 3.40 (s, 2H), 3.30-3.20 (m, 2H), 2.99-2.92 (m,
2H), 2.50-2.31 (m, 2H), 2.21-2.13 (m, 2H), 1.74-1.62 (m, 4H),
1.61-1.36 (m, 4H).
Reference Example 23
[0374] Reactions and treatments were carried out in the same manner
as Reference example 22 using the corresponding starting material
compounds, thereby givining the compounds of Reference example 23
shown in Table 3.
TABLE-US-00003 TABLE 3 Reference Example structure Spectral data 23
##STR00093## 300 MHz .sup.1H-NMR (DMSO-d.sub.6, .delta.) 10.24 (s,
1H), 7.06 (s, 1H), 7.00 (d, 1H, J = 8.1 Hz), 6.69 (d, 1H, J = 8.1
Hz), 4.42-4.34 (m, 1H), 3.91-3.77 (m, 2H), 3.72-3.56 (m, 2H), 3.40
(s, 2H), 3.16-3.07 (m, 1H), 3.01-2.86 (m, 4H), 2.45-2.30 (m, 1H),
2.27-2.16 (m, 2H), 1.96 (s, 3H), 1.79-1.45 (m, 3H), 1.45-1.13 (m,
2H).
Reference Example 24
Production of 5-(1-(pyrimidin-2-yl)piperidin-4-yl)indolin-2-one
##STR00094##
[0376] To a solution of 5-(piperidin-4-yl)indolin-2-one (39.8 mg,
0.184 mmol) in EtOH 3 ml) were added 2-chloropyrimidine (33.7 mg,
0.294 mmol) and iPr.sub.2NEt (0.095 ml, 0.551 mmol). After stirring
at 80.degree. C. for 6 h, the reaction mixture was concentrated.
The residue was purified by column chromatography (CHCl.sub.3/MeOH)
to give 5-(1-(pyrimidin-2-yl)piperidin-4-yl)indolin-2-one (47.9 mg,
88%).
[0377] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 8.35 (s, 1H), 8.33
(s, 1H), 7.54 (brs, 1H), 7.08 (s, 1H), 7.04 (d, 1H, J=7.9 Hz), 6.77
(d, 1H, J=7.9 Hz), 6.58-6.47 (m, 1H), 4.99-4.90 (m, 2H), 3.49 (s,
2H), 3.03-2.91 (m, 2H), 2.81-2.69 (m, 1H), 1.98-1.88 (m, 2H),
1.73-1.50 (m, 2H).
Reference Example 25
Production of 5-(1-phenylpiperidin-4-yl)indolin-2-one
##STR00095##
[0379] To a solution of LHMDS (3.2 ml, 1.10 M in hexane, 3.52 mmol)
in THF (30 ml) was added a solution of 1-phenylpiperidin-4-one (559
mg, 3.19 mmol) in THF (7 ml) at -78.degree. C. over 3 min. After
stirring at the same temperature for 30 min, PhNTf.sub.2 (1.48 g,
4.15 mmol) was added. After stirring at -78.degree. C. for 20 min,
then the reaction mixture was stirred at 0.degree. C. for 20 min.
The reaction mixture was quenched by sat. NH.sub.4Cl aq. and
extracted with CHCl.sub.3. The organic layer was dried over
Na.sub.2SO.sub.4 and concentrated. The residue was purified by
column chromatography (hexane/EtOAc) to give
1-phenyl-1,2,3,6-tetrahydropyridin-4-yl trifluoromethanesulfonate
(563 mg, 58%).
[0380] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.32-7.24 (m, 2H),
6.97-6.87 (m, 3H), 5.90-5.86 (m, 1H), 3.87-3.82 (m, 2H), 3.50 (t,
2H, J=5.6 Hz), 2.62-2.56 (m, 2H).
[0381] To a solution of 1-phenyl-1,2,3,6-tetrahydropyridin-4-yl
trifluoromethanesulfonate (104 mg, 0.339 mmol) in 1,4-Dioxane (3
ml) and H.sub.2O (1 ml) were added
5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indolin-2-one (97.6
mg, 0.377 mmol), Pd(PPh.sub.3).sub.4 (38.8 mg, 0.00336 mmol), LiCl
(47.1 mg, 1.11 mmol) and K.sub.2CO.sub.3 (140 mg, 1.01 mmol). After
stirring at 120.degree. C. in microwave reactor for 1 h, the
reaction mixture was quenched by sat. NaHCO.sub.3 aq. The resulting
mixture was extracted with CHCl.sub.3, the organic layer was washed
with brine, dried over Na.sub.2SO.sub.4 and concentrated. The
residue was purified by column chromatography (CHCl.sub.3/MeOH) to
give 5-(1-phenyl-1,2,3,6-tetrahydropyridin-4-yl)indolin-2-one (75.8
mg) as mixture of triphenylphosphine oxide.
[0382] MS m/z 291 (M+H)
[0383] To a solution of
5-(1-phenyl-1,2,3,6-tetrahydropyridin-4-yl)indolin-2-one (75.8 mg,
0.261 mmol) in THF (3 ml) and MeOH (3 ml) was added 10% Pd/C (210
mg) and stirred at room temperature under H.sub.2 (1 atom)
atmosphere for 2 h. The reaction mixture was filtered through a
Celite pad and concentrated. The residue was purified by column
chromatography (CHCl.sub.3/MeOH) to afford
5-(1-phenylpiperidin-4-yl)indolin-2-one (51.2 mg) as mixture of
triphenylphosphine oxide.
[0384] MS m/z 293 (M+H)
Reference Example 26
Production of
(Z)--N-(2-(diethylamino)ethyl)-2,4-dimethyl-5-((2-oxo-5-(4,4,
5,5-tetramethyl-1,3,2-dioxab
orolan-2-yl)indolin-3-ylidene)methyl)-1H-pyrrole-3-carboxamide
##STR00096##
[0386] To a solution of
5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indolin-2-one (195
mg, 0.75 mmol) in EtOH (3 ml) was added
N-(2-(diethylamino)ethyl)-5-formyl-2,4-dimethyl-1H-pyrrole-3-carboxamide
(200 mg, 0.76 mmol) and piperidine (82 .mu.L, 0.83 mmol). The
mixture was stirred at 80.degree. C. for 1 hour. After cooled down
to room temperature, the reaction mixture was concentrated,
filtrated, and washed with EtOH to give
(Z)--N-(2-(diethylamino)ethyl)-2,4-dimethyl-5-((2-oxo-5-(4,4,
5,5-tetramethyl-1,3,2-dioxab
orolan-2-yl)indolin-3-ylidene)methyl)-1H-pyrrole-3-carboxamide (218
mg) as yellow solid.
[0387] MS m/z 507.6 (M+H)
Reference Example 27
Production of
(Z)-5-((5-bromo-2-oxoindolin-3-ylidene)methyl)-N-(2-(diethylamino)ethyl)--
2,4-dimethyl-1H-pyrrole-3-carboxamide
##STR00097##
[0389] To the solution of 5-bromoindolin-2-one (262 mg, 1.24 mmol)
in EtOH (5 ml) was added
N-(2-(diethylamino)ethyl)-5-formyl-2,4-dimethyl-1H-pyrrole-3-carboxamide
(298 mg, 1.12 mmol) and piperidine (112 .mu.L, 1.13 mmol). The
mixture was stirred at 80.degree. C. for 1 hour. After cooled down
to room temperature, the reaction mixture was concentrated,
filtrated, and washed with EtOH to give
(Z)-5-((5-bromo-2-oxoindolin-3-ylidene)methyl)-N-(2-(diethylamino)ethyl)--
2,4-dimethyl-1H-pyrrole-3-carboxamide (368 mg) as orange solid.
[0390] MS m/z 459.4/461.4 (M+H)
Example 1
Production of
(Z)--N-(2-(diethylamino)ethyl)-2,4-dimethyl-5-((2-oxo-5-(5-phenylthiophen-
-2-)indolin-3-ylidene)methyl)-1H-pyrrole-3-carboxamide
##STR00098##
[0392] To a solution of No (23 mg, 0.079 mmol) in THF/EtOH (1 ml/1
ml) was added No (25.2 mg, 0.095 mmol) and piperidine (0.7 mg,
0.008 mmol). The mixture was stirred at 80.degree. C. for 10 hours.
After cooled down to the room temperature, the reaction mixture was
concentrated, filtrated, and washed with EtOH to give
(Z)--N-(2-(diethylamino)ethyl)-2,4-dimethyl-5-((2-oxo-5-(5-phenylthiophen-
-2-yl)indolin-3-ylidene)methyl)-1H-pyrrole-3-carboxamide (18 mg) as
an orange solid.
[0393] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 13.67 (s, 1H),
11.02 (s, 1H), 8.31 (s, 1H), 7.81 (s, 1H), 7.70-7.76 (m, 2H), 7.51
(s, 2H), 7.42-7.45 (m, 4H), 7.32-7.39 (m, 1H), 6.90-6.93 (m, 1H),
3.25-3.34 (m, 4H), 2.4-2.6 (m, 10H), 0.94-0.99 (m, 6H); MS m/z
539.70 (M+H).
Examples 2 to 50
[0394] Reactions and treatments were carried out in the same manner
as in Example 1 using the corresponding starting material
compounds, thereby giving the compounds of Examples 2 to 50 shown
in Table 4.
TABLE-US-00004 TABLE 4 Ex- am- ple structure Spectral data 2
##STR00099## 30 0MHz .sup.1H-NMR (CDCl.sub.3, .delta.) 13.29 (s,
1H), 8.48 (s, 1H), 8.19 (d, 1H), 7.99-7.96 (m, 1H), 7.66 (dd, 1H, J
= 8.4, 1.8 Hz), 7.48-7.45 (m, 5H), 6.95 (d, 1H, J = 8.1 Hz),
3.74-3.47 (brs, 4H), 2.38-2.24 (m, 13H) 3 ##STR00100## 300 MHz
.sup.1H-NMR (DMSO-d.sub.6, .delta.) 13.62 (s, 1H), 11.02 (s, 1H),
8.17 (s, 1H), 7.80 (s, 1H), 7.70-7.68 (m, 2H), 7.55-7.51 (m, 2H),
7.45-7.41 (m, 3H), 7.32-7.29 (m, 1H), 6.94-6.92 (m, 1H), 3.47-3.42
(m, 4H), 2.49 (s, 3H), 2.32 (s, 3H), 2.30-2.27 (m, 4H), 2.19 (s,
3H) 4 ##STR00101## 300 MHz .sup.1H-NMR (CDCl.sub.3, .delta.) 13.29
(s, 1H), 8.53 (s, 1H), 7.67 (d, 2H, J = 8.4 Hz), 7.50 (s, 1H), 7.40
(t, 2H, J = 7.8 Hz), 7.30-7.23 (m, 3H), 6.83 (d, 1H, J = 7.8 Hz),
6.70 (brs, 1H), 3.48 (d, 2H, J = 5.4 Hz), 2.68-2.57 (m, 6H), 2.49
(s, 3H), 2.39 (s, 3H), 1.02 (t, 6H, J = 6.9 Hz) 5 ##STR00102## 300
MHz .sup.1H-NMR (CDCl.sub.3, .delta.) 13.63 (s, 1H), 8.14 (s, 1H),
7.78 (s, 1H), 7.45-7.42 (m, 4H), 7.40-7.36 (m, 2H), 7.24-7.17 (m,
2H), 6.95-6.92 (m, 1H), 3.74-3.72 (m, 4H), 2.63-2.56 (m, 4H), 2.42
(s, 3H), 2.38 (s, 3H), 2.32 (s, 3H), 2.30 (s, 3H) 6 ##STR00103##
300 MHz .sup.1H-NMR (CDCl.sub.3, .delta.) 13.34 (s, 1H), 8.15 (s,
1H), 7.98 (s, 1H), 7.73 (d, 2H, J = 7.5 Hz), 7.60 (s, 1H),
7.46-7.41 (m, 3H), 7.32 (t, 2H, J = 7.5 Hz), 6.90 (d, 1H, J = 7.8
Hz), 3.69 (brs, 4H), 2.42-2.33 (m, 14H) 7 ##STR00104## 300 MHz
.sup.1H-NMR (DMSO-d.sub.6, .delta.) 13.63 (s, 1H), 11.01 (s, 1H),
8.15 (s, 1H), 8.05-8.00 (m, 2H), 7.87 (d, 1H, J = 8.1 Hz),
7.72-7.68 (m, 2H), 7.58-7.50 (m, 2H), 7.46-7.41 (m, 1H), 3.38-3.25
(m, 4H), 2.60-2.47 (m, 4H), 2.45 (s, 3H), 2.43 (s, 3H), 0.98 (t,
6H, J = 7.0 Hz) 8 ##STR00105## 300 MHz .sup.1H-NMR (DMSO-d.sub.6,
.delta.) 13.57 (s, 1H), 11.01 (s, 1H), 8.15 (s, 1H), 8.04-8.00 (m,
2H), 7.86 (d, 1H, J = 8.1 Hz), 7.70 (dd, 1H, J = 1.5, 6.6 Hz), 7.66
(s, 1H), 7.58-7.51 (m, 5H), 3.55-3.35 (m, 4H), 2.37-2.23 (m, 4H),
2.29 (s, 3H), 2.27 (s, 3H), 2.18 (s, 3H) 9 ##STR00106## 300 MHz
.sup.1H-NMR (DMSO-d.sub.6, .delta.) 13.65 (s, 1H), 10.41 (s, 1H),
8.24 (s, 1H), 8.08-8.04 (m, 2H), 7.87 (d, 1H, J = 7.5 Hz), 7.75 (s,
1H), 7.72 (d, 1H, J = 8.3 Hz), 7.62-7.54 (m, 3H), 7.47-7.43 (m,
1H), 7.14 (dd, 1H, J = 7.7, 7.9 Hz), 3.36-3.23 (m, 4H), 2.56-2.45
(m, 2H), 2.46 (s, 3H), 2.44 (s, 3H), 0.97 (t, 6H, J = 7.1 Hz) 10
##STR00107## 300 MHz .sup.1H-NMR (DMSO-d.sub.6, .delta.) 13.60 (s,
1H), 10.41 (s, 1H), 8.25 (s, 1H), 8.08-8.05 (m, 2H), 7.86 (d, 1H, J
= 7.7 Hz), 7.73 (s, 1H), 7.71 (d, 1H, J = 8.8 Hz), 7.62-7.54 (m,
3H), 7.14 (t, 1H, J = 7.6 Hz), 3.57-3.37 (m, 4H), 2.32-2.25 (m,
4H), 2.30 (s, 3H), 2.28 (s, 3H), 2.18 (s, 3H) 11 ##STR00108## 300
MHz .sup.1H-NMR (DMSO-d.sub.6, .delta.) 13.62 (s, 1H), 11.02 (s,
1H), 8.17 (s, 1H), 7.80 (s, 1H), 7.70-7.67 (m, 2H), 7.53-7.49 (m,
2H), 7.49-7.42 (m, 3H), 7.32-7.30 (m, 1H), 6.94-6.91 (m, 1H),
3.60-3.30 (m, 8H), 2.30 (s, 3H), 2.28 (s, 3H) 12 ##STR00109## 300
MHz .sup.1H-NMR (DMSO-d.sub.6, .delta.) 13.66 (s, 1H), 10.77 (s,
1H), 7.75 (s, 1H), 7.67 (s, 1H), 7.38 (t, 1H, J = 5.5 Hz), 6.96
(brd, 1H, J = 7.9 Hz), 6.75 (d, 1H, J = 7.9 Hz), 3.92-3.85 (m, 2H),
3.34-3.22 (m, 6H), 3.03-2.97 (m, 2H), 2.57-2.39 (m, 6H), 2.42 (s,
3H), 2.42 (s, 3H), 2.23-2.12 (m, 2H), 1.76-1.65 (m, 6H), 1.52-1.36
(m, 2H), 0.97 (t, 6H, J = 7.2 Hz) 13 ##STR00110## 300 MHz
.sup.1H-NMR (CDCl.sub.3, .delta.) 13.35 (s, 1H), 8.14-8.11 (m, 1H),
7.74 (s, 1H), 7.51-7.45 (m, 4H), 7.42-7.38 (m, 2H), 7.29-7.21 (m,
2H), 6.96-6.90 (m, 1H), 3.80-3.40 (m, 8H), 2.43 (s, 3H), 2.40 (s,
3H), 2.24 (s, 3H) 14 ##STR00111## 300 MHz .sup.1H-NMR
(DMSO-d.sub.6, .delta.) 13.66 (s, 1H), 11.02 (s, 1H), 8.19 (s, 1H),
7.81 (s, 1H), 7.70-7.67 (m, 2H), 7.45-7.42 (m, 2H), 7.40-7.32 (m,
3H), 7.30-7.27 (m, 1H), 6.93-6.91 (m, 1H), 3.60-3.30 (m, 4H),
2.71-2.65 (m, 4H), 2.50 (s, 3H), 2.49 (s, 3H), 1.70-1.60 (m, 4H) 15
##STR00112## LCMS m/z 539.70 (M + H) 16 ##STR00113## 300 MHz
.sup.1H-NMR (CDCl.sub.3, .delta.) 13.37 (s, 1H), 8.70 (s, 1H), 7.77
(s, 1H), 7.49-7.47 (m, 4H), 7.40-7.36 (m ,2H), 7.20-7.17 (m, 1H),
6.94-6.85 (m, 2H), 3.58-3.56 (m, 2H), 2.78-2.71 (m, 6H), 2.56 (s,
3H), 2.42 (s, 3H), 2.41 (s, 3H), 1.12-1.08 (m, 6H) 17 ##STR00114##
400 MHz .sup.1H-NMR (DMSO-d.sub.6, .delta.) 13.67 (s, 1H), 1.11
(br, 1H), 8.29 (d, 1H), J = 1.5 Hz), 8.15-8.10 (m, 2H), 7.83 (s,
1H), 7.73 (s, 1H), 7.65-7.43 (m, 5H), 7.00 (d, 1H, J = 8.1 Hz),
3.35-3.26 (m, 4H), 2.58-2.45 (m, 10H), 0.98 (t, 6H, J = 7.1 Hz) 18
##STR00115## 400 MHz .sup.1H-NMR (DMSO-d.sub.6, .delta.) 13.61 (s,
1H), 11.10 (br, 1H), 7.81 (s, 1H), 7.73 (s, 1H), 7.65-7.51 (m, 4H),
7.00 (d, 1H, J = 8.1 Hz), 3.7-3.3 (m, 4H), 2.35-2.22 (m, 4H), 2.34
(s, 3H), 2.31 (s, 3H), 2.19 (s, 3H) 19 ##STR00116## 400 MHz
.sup.1H-NMR (CDCl.sub.3, .delta.) 13.35 (s, 1H), 8.36 (br, 1H),
8.20 (d, 1H, J = 1.4 Hz), 7.93 (dd, 1H, J = 8.1, 1.4 Hz), 7.75-7.70
(m, 2H), 7.52 (s, 1H), 7.48-7.30 (m, 4H), 6.99 (d, 1H, J = 8.1 Hz),
6.62 (br, 1H), 3.52 (m, 2H), 2.70 (t, 2H, J = 5.8 Hz), 2.62 (q, 4H,
J = 7.1 Hz), 2.50 (s, 3H), 2.48 (s, 3H), 1.06 (t, 6H, J = 7.1 Hz)
20 ##STR00117## 400 MHz .sup.1H-NMR (CDCl.sub.3, .delta.) 13.32 (s,
1H), 8.22 (d, 1H, J = 1.6 Hz), 8.14 (br, 1H), 7.93 (dd, 1H, J =
8.1, 1.6 Hz), 7.76-7.72 (m, 2H), 7.52 (s, 1H), 7.49-7.32 (m, 4H),
7.00 (d, 1H, J = 8.1 Hz), 4.0-3.3 (m, 4H), 2.6-2.3 (m, 4H), 2.40
(s, 3H), 2.34 (s, 3H), 2.33 (s, 3H) 21 ##STR00118## 400 MHz
.sup.1H-NMR (CDCl.sub.3, .delta.) 13.34 (s, 1H), 8.24 (br, 1H),
7.96-7.91 (m, 2H), 7.77-7.72 (m, 2H), 7.53 (s, 1H), 7.49-7.32 (m,
4H), 7.00 (d, 1H, J = 8.1 Hz), 4.0-3.3 (m, 8H), 2.41 (s, 3H), 2.36
(s, 3H) 22 ##STR00119## 400 MHz .sup.1H-NMR (CDCl.sub.3, .delta.)
13.26 (s, 1H), 7.69 (s, 1H), 7.67 (d, 1H, J = 1.8 Hz), 7.30-7.23
(m, 6H), 7.17 (s, 1H), 6.90 (dd, 1H, J = 1.6, 8.1 Hz), 6.73 (d, 1H,
J = 8.1 Hz), 6.47 (d, 1H, J = 1.8 Hz), 3.55-3.40 (m, 2H), 2.73-2.47
(m, 6H), 2.53 (s, 3H), 2.39 (s, 3H), 1.09-0.91 (m, 6H). 23
##STR00120## 400 MHz .sup.1H-NMR (CDCl.sub.3, .delta.) 13.27 (s,
1H), 8.05 (s, 1H), 7.76 (d, 1H, J = 1.8 Hz), 7.39-7.31 (m, 6H),
7.21 (s, 1H), 6.99 (dd, 1H, J = 1.6, 8.1 Hz), 6.81 (d, 1H, J = 8.1
Hz), 6.55 (d, 1H, J = 1.8 Hz), 3.95-3.37 (br, 4H), 2.56-2.30 (m,
4H), 2.41 (s, 3H), 2.35 (s, 3H), 2.27 (s, 3H). 24 ##STR00121## 400
MHz .sup.1H-NMR (CDCl.sub.3, .delta.) 13.22 (s, 1H), 7.84 (s, 1H),
7.71 (d, 1H, J = 1.8 Hz), 7.55 (dt, 2H, J = 1.9, 8.7 Hz), 7.41 (dt,
2H, J = 1.9, 8.7 Hz), 7.35 (1H, d, J = 1.5 Hz), 7.22 (s, 1H), 6.84
(dd, 1H, J = 1.5, 8.0 Hz), 6.78 (d, 1H, J = 8.0 Hz), 6.48 (d, 1H, J
= 1.8 Hz), 3.90-3.30 (br, 4H), 2.47-2.20 (m, 4H), 2.33 (s, 3H),
2.27 (s, 3H), 2.17 (s, 3H). 25 ##STR00122## 400 MHz .sup.1H-NMR
(CDCl.sub.3, .delta.) 13.30 (s, 1H), 7.96 (s, 1H), 7.71 (d, 1H, J =
1.8 Hz), 7.55 (dt, 2H, J = 2.1, 8.7 Hz), 7.41 (dt, 2H, J = 2.1, 8.7
Hz), 7.36 (1H, d, J = 1.5 Hz), 7.26 (s, 1H), 6.83 (dd, 1H, J = 1.5,
8.0 Hz), 6.78 (d, 1H, J = 8.0 Hz), 6.48 (d, 1H, J = 1.8 Hz),
3.47-3.40 (m, 2H), 2.65-2.49 (m, 6H), 2.53 (s, 3H), 2.38 (s, 3H),
0.98 (t, 6H, J = 6.9 Hz). 26 ##STR00123## 300 MHz .sup.1H-NMR
(CDCl.sub.3, .delta.) 13.38 (s, 1H), 8.86 (s, 1H), 8.48 (d, 1H, J =
4.5 Hz), 8.21 (s, 1H), 7.84 (d, 1H, J = 7.5 Hz), 7.63 (s, 1H),
7.43-7.34 (m, 2H), 7.30-7.25 (m, 2H), 6.92 (d, 1H, J = 8.1 Hz),
3.72 (brs, 2H), 3.03-2.92 (m, 6H), 2.59 (s, 3H), 2.52 (s, 3H), 1.26
(d, 6H, J = 5.1 Hz) 27 ##STR00124## 400 MHz .sup.1H-NMR
(CDCl.sub.3, .delta.) 13.33 (s, 1H), 7.80 (s, 1H), 7.60 (d, 1H, J =
1.6 Hz), 7.40 (s, 1H), 7.35 (dd, 1H, J = 1.7, 8.1 Hz), 7.16-7.12
(m, 3H), 7.08 (d, 1H, J = 3.7 Hz), 6.97 (dd, 1H, J = 3.7, 5.1 Hz),
6.84 (d, 1H, J = 8.1 Hz), 3.69-3.63 (m, 2H), 3.54-3.40 (m, 2H),
2.72-2.49 (m, 4H), 2.54 (s, 3H), 2.47 (s, 3H), 1.08-0.95 (m, 6H).
28 ##STR00125## 300 MHz .sup.1H-NMR (CDCl.sub.3, .delta.) 13.22 (s,
1H), 9.63 (s, 1H), 7.76 (s, 1H), 7.46-7.37 (m, 5H), 7.19-7.11 (m,
3H), 6.90-6.88 (m, 1H), 3.69-3.60 (m, 2H), 3.05-2.80 (m, 6H), 2.51
(s, 3H), 2.41 (s, 3H), 2.21 (s, 3H), 2.02-1.91 (m, 4H) 29
##STR00126## LCMS m/z 523.66 (M + H) 30 ##STR00127## 300 MHz
.sup.1H-NMR (CDCl.sub.3, .delta.) 13.34 (s, 1H), 8.82 (s, 1H), 8.43
(d, 1H, J = 4.2 Hz), 8.01 (t, 2H, J = 3.6 Hz), 7.86-7.81 (m, 2H),
7.67 (s, 1H), 7.42- 7.37 (m, 2H), 7.20 (t, 1H, J = 6.3 Hz), 6.91
(d, 1H, J = 7.8 Hz), 3.69 (brs, 8H), 2.89 (s, 3H), 2.36 (s, 3H) 31
##STR00128## 300 MHz .sup.1H-NMR (DMSO-d.sub.6, .delta.) 13.61 (s,
1H), 10.93 (s, 1H), 9.09 (s, 1H), 8.50 (d, 1H, J = 4.8 Hz), 8.32
(s, 1H), 8.25 (s, 1H), 8.00-7.95 (m, 2H), 7.78 (s, 1H), 7.54 (d,
1H, J = 8.4 Hz), 7.37 (t, 1H, J = 5.4 Hz), 6.89 (d, 1H, J = 7.8
Hz), 3.47 (brs, 4H), 2.32-2.26 (m, 13H) 32 ##STR00129## 300 MHz
.sup.1H-NMR (CDCl.sub.3, .delta.) 13.38 (s, 1H), 8.79 (s, 1H), 8.42
(d, 1H, J = 3.6 Hz), 8.11 (s, 1H), 8.00-7.97 (m, 2H), 7.81 (t, 1H,
J = 6.9 Hz), 7.64 (s, 1H), 7.42 (s, 1H), 7.19 (d, 1H, J = 6.6 Hz),
6.91 (d, 1H, J = 7.8 Hz), 3.56 (brs, 4H), 2.79-2.70 (m, 6H), 2.58
(s, 3H), 2.51 (s, 3H). 33 ##STR00130## 300 MHz .sup.1H-NMR
(CDCl.sub.3, .delta.) 13.34 (s, 1H), 8.78 (s, 1H), 8.40 (d, 1H, J =
3.9 Hz), 8.30 (s, 1H), 7.99-7.95 (m, 2H), 7.80 (t, 1H, J = 7.5 Hz),
7.62 (s, 1H), 7.38-7.35 (m, 2H), 7.18 (d, 1H, J = 7.2 Hz), 6.89 (d,
1H, J = 7.8 Hz), 3.70 (brs, 2H), 3.02 (brs, 6H), 2.56 (s, 3H), 2.47
(s, 3H), 1.96 (brs, 4H) 34 ##STR00131## 300 MHz .sup.1H-NMR
(DMSO-d.sub.6, .delta.) 13.71 (s, 1H), 11.09 (s, 1H), 8.10-8.07 (m,
3H), 7.82 (s, 1H), 7.53-7.45 (m, 5H), 7.03-7.00 (m, 1H), 3.34-3.27
(m, 7H), 2.45-2.39 (m, 10H), 0.99-0.94 (m, 6H) 35 ##STR00132## 300
MHz .sup.1H-NMR (DMSO-d.sub.6, .delta.) 13.64 (s, 1H), 11.86 (s,
1H), 8.12-8.07 (m, 2H), 7.81 (s, 1H), 7.58-7.48 (m ,5H), 7.09-7.03
(m, 1H), 3.96-3.45 (m, 7H), 2.50-2.47 (m, 4H), 2.34 (s, 3H), 2.27
(s, 3H), 2.14 (s, 3H) 36 ##STR00133## 300 MHz .sup.1H-NMR
(DMSO-d.sub.6, .delta.) 13.59 (s, 1H), 11.02 (s, 1H), 8.15 (s, 1H),
8.05-8.00 (m, 2H), 7.87 (d, 1H, J = 8.3 Hz), 7.72-7.67 (m, 2H),
7.58-7.53 (m, 4H), 3.65-3.53 (m, 4H), 2.31 (s, 3H), 2.29 (s, 3H) 37
##STR00134## LCMS m/z 538.65 (M + H) 38 ##STR00135## LCMS m/z
522.61 (M + H) 39 ##STR00136## LCMS m/z 536.64 (M + H) 40
##STR00137## 300 MHz .sup.1H-NMR (DMSO-d.sub.6, .delta.) 13.60 (s,
1H), 10.76 (s, 1H), 7.73 (s, 1H), 7.65 (s, 1H), 6.96 (dd, 1H, J =
7.9, 8.4 Hz), 6.75 (d, 1H, J = 7.9 Hz), 3.92-3.84 (m, 2H),
3.57-3.35 (m, 4H), 3.04-2.96 (m, 2H), 2.52-2.35 (m, 3H), 2.32-2.15
(m, 7H), 2.26 (s, 3H), 2.26 (s, 3H), 2.18 (s, 3H), 1.78-1.64 (m,
6H), 1.52-1.37 (m, 2H) 41 ##STR00138## 300 MHz .sup.1H-NMR
(DMSO-d.sub.6, .delta.) 13.68 (s, 1H), 12.61 (s, 1H), 10.93 (s,
1H), 8.18 (s, 1H), 8.03-8.01 (m, 2H), 7.78-7.70 (m, 2H), 7.51-7.44
(m, 2H), 7.32-7.37 (m, 1H), 6.91-6.89 (m, 1H), 3.34-3.30 (m, 4H),
2.64-2.60 (m, 4H), 2.50-2.48 (m, 6H), 0.99-0.96 (m, 6H) 42
##STR00139## 300 MHz .sup.1H-NMR (DMSO-d.sub.6, .delta.) 13.68 (s,
1H), 10.80 (s, 1H), 7.78 (s, 1H), 7.68 (s, 1H), 7.24-7.19 (m ,2H),
7.04-6.95 (m, 3H), 6.80-6.74 (m, 2H), 3.84-3.76 (m, 2H), 3.35-3.22
(m, 3H), 2.78-2.53 (m, 6H), 2.50-2.36 (m, 2H), 2.43 (s, 3H), 2.42
(s, 3H), 1.91-1.81 (m, 4H), 1.01 (brs, 6H) 43 ##STR00140## 300 MHz
.sup.1H-NMR (DMSO-d.sub.6, .delta.) 13.66 (s, 1H), 10.79 (s, 1H),
8.36 (s, 1H), 8.34 (s, 1H), 7.73 (s, 1H), 7.65 (s, 1H), 7.40-7.36
(m, 1H), 6.99 (brd, 1H, J = 7.9 Hz), 6.77 (d, 1H, J = 7.9 Hz), 6.58
(t, 1H, J = 4.6 Hz), 4.90-4.80 (m, 2H), 3.30-3.20 (m, 2H),
2.98-2.86 (m, 2H), 2.86-2.69 (m, 1H), 2.56-2.44 (m, 6H), 2.42 (s,
3H), 2.40 (s, 3H), 1.89- 1.78 (m, 2H), 1.72-1.55 (m, 2H), 0.96 (t,
6H, J = 7.0 Hz) 44 ##STR00141## 300 MHz .sup.1H-NMR (DMSO-d.sub.6,
.delta.) 13.62 (s, 1H), 12.60 (s, 1H), 10.90 (s, 1H), 8.14 (s, 1H),
8.02-7.99 (m, 2H), 7.71-7.67 (m, 2H), 7.51-7.44 (m, 2H), 7.37-7.32
(m, 1H), 6.90-6.87 (m, 1H), 3.45-3.33 (m, 4H), 2.50-2.48 (m, 4H),
2.29 (s, 3H), 2.19 (s, 3H), 2.14 (s, 3H) 45 ##STR00142## 300 MHz
.sup.1H-NMR (DMSO-d.sub.6, .delta.) 13.66 (s, 1H), 10.77 (s, 1H),
7.75 (s, 1H), 7.67 (s, 1H), 7.38 (t, 1H, J = 5.6 Hz), 6.96 (brd,
1H, J = 7.9 Hz), 6.75 (d, 1H, J = 7.9 Hz), 4.43-4.36 (m, 1H),
3.86-3.80 (m, 1H), 3.35-3.23 (m, 4H), 2.98-2.93 (m, 2H), 2.55- 2.38
(m, 8H), 2.42 (s, 3H), 2.42 (s, 3H), 2.30-2.15 (m, 2H), 1.98 (s,
3H), 1.70-1.64 (m, 6H), 1.48-1.15 (m, 2H), 0.96 (t, 6H, J = 7.2 Hz)
46 ##STR00143## 300 MHz .sup.1H-NMR (DMSO-d.sub.6, .delta.) 13.60
(s, 1H), 10.77 (s, 1H), 7.73 (s, 1H), 7.65 (s, 1H), 6.96 (d, 1H, J
= 7.9 Hz), 6.75 (brd, 1H, J = 7.9 Hz), 4.43-4.35 (m, 1H), 3.88-3.78
(m, 1H), 3.60-3.38 (m, 4H), 3.04-2.90 (m, 2H), 2.56-2.20 (m, 8H),
2.26 (s, 3H), 2.26 (s, 3H), 2.19 (s, 3h), 2.00-1.91 (m, 2H), 1.98
(s, 3H), 1.70-1.65
(m, 6H), 1.48-1.18 (m, 2H) 47 ##STR00144## 300 MHz .sup.1H-NMR
(DMSO-d.sub.6, .delta.) 13.64 (s, 1H), 10.88 (s, 1H), 8.19 (s, 1H),
8.00 (s, 1H), 7.90 (s, 1H), 7.68 (s, 1H), 7.45-7.38 (m, 1H), 7.35
(d, 1H, J = 7.9 Hz), 6.85 (d, 1H, J = 7.9 Hz), 4.44-4.33 (m, 1H),
4.02-3.91 (m, 2H), 3.52-3.43 (m, 2H), 3.32-3.24 (m, 2H), 2.58-2.48
(m, 6H), 2.44 (s, 3H), 2.44 (s, 3H), 2.06- 1.89 (m, 4H), 0.97 (t,
6H, J = 7.0 Hz) 48 ##STR00145## 300 MHz .sup.1H-NMR (DMSO-d.sub.6,
.delta.) 13.64 (s, 1H), 11.06 (s, 1H), 9.18 (s, 1H), 8.30 (s, 1H),
7.96 (d, 2H, J = 9 Hz), 7.75-7.45 (m, 6H), 7.00 (d, 1H, J = 9 Hz),
3.39-3.26 (m, 4H), 2.56-2.45 (m, 10H), and 0.97 (t, J = 7.5 Hz). 49
##STR00146## LCMS m/z 524.5 (M + H) 50 ##STR00147## 300 MHz
.sup.1H-NMR (DMSO-d.sub.6, .delta.) 13.64 (s, 1H), 11.14 (s, 1H),
8.41 (s, 1H), 8.12-8.11 (m, 2H), 7.87-7.85 (m, 1H), 7.84 (s, 1H),
7.55-7.48 (m, 4H), 7.03-7.01 (m, 1H), 3.34-3.25 (m, 4H), 2.54-2.45
(m, 10H), 0.99-0.95 (m, 6H)
Example 51
Production of
(Z)--N-(2-(diethylamino)ethyl)-2,4-dimethyl-5-((2-oxo-5-(4-phenylthiazol--
2-yl)indolin-3-ylidene)methyl)-1H-pyrrole-3-carboxamide
##STR00148##
[0396] To a solution of No (40 mg, 0.079 mmol) in DMF/H.sub.2O (3
ml/1 ml) was added Pd(PPh.sub.3).sub.4 (9.1 mg, 0.008 mmol),
5-phenylthiophene-2-boronic acid (23 mg, 0.095 mmol) and potassium
carbonate (33 mg, 0.237 mmol). The mixture was stirred at
110.degree. C. for 1 hour under microwave irradiation. The mixture
was extracted with CHCl.sub.3, and the organic layer was washed
with H.sub.2O, dried over Na.sub.2SO.sub.4 and concentrated in
vacuo. The residue was purified by column chromatography
(CHCl.sub.3/MeOH) to give
(Z)--N-(2-(diethylamino)ethyl)-2,4-dimethyl-5-((2-oxo-5-(4-phenylthiazol--
2-yl)indolin-3-yl idene)methyl)-1H-pyrrole-3-carboxamide (10 mg) as
yellow solid.
[0397] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 13.70 (s, 1H),
11.20 (s, 1H), 8.43 (s, 1H), 8.07-8.11 (m, 3H), 7.79-7.90 (m. 2H),
7.39-7.50 (m, 4H), 7.02 (m, 1H), 3.25-3.35 (m, 4H), 2.4-2.6 (m,
10H), 0.95-0.99 (m, 6H); MS m/z 540.69 (M+H).
Examples 52 to 55
[0398] Reactions and treatments were carried out in the same manner
as in Example 1 using the corresponding starting material
compounds, thereby giving the compounds of Examples 52 to 55 shown
in Table 5.
TABLE-US-00005 TABLE 5 Ex- am- ple structure Spectral data 52
##STR00149## 300 MHz .sup.1H-NMR (CDCl.sub.3, .delta.) 13.24 (s,
1H), 8.03 (s, 1H), 7.89 (t, 1H, J = 3.3 Hz), 7.83 (s, 1H), 7.58 (t,
1H, J = 9.0 Hz), 7.41-7.27 (m, 4H), 6.91-6.82 (m, 1H), 3.62 (brs,
2H), 2.93-2.82 (m, 6H), 2.62-2.36 (m, 6H), 1.18-1.15 (m, 6H) 53
##STR00150## 300 MHz .sup.1H-NMR (CD.sub.3OD, .delta.) 8.34 (s,
1H), 8.16 (d, 2H, J = 7.8 Hz), 7.92 (d, 1H, J = 8.4 Hz), 7.65-7.60
(m, 5H), 7.08 (d, 1H, J = 8.1 Hz), 3.59 (t, 2H, J = 6.6 Hz),
2.99-2.88 (m, 6H), 2.52 (s, 3H), 2.50 (s, 3H), 1.22 (t, 6H, J = 7.2
Hz) 54 ##STR00151## 300 MHz .sup.1H-NMR (DMSO-d.sub.6, .delta.)
13.64 (s, 1H), 11.07 (s, 1H), 8.28 (s, 1H), 8.23 (s, 1H), 7.96-7.94
(m, 2H), 7.83 (s, 1H), 7.55-7.42 (m ,4H), 6.97-6.94 (m, 1H),
3.38-3.25 (m, 4H), 2.56-2.40 (m, 10H), 0.99-0.95 (m, 6H) 55
##STR00152## 300 MHz .sup.1H-NMR (DMSO-d.sub.6, .delta.) 13.65 (s,
1H), 11.25 (s, 1H), 7.86 (s, 1H), 7.75 (s, 1H), 7.55 (s, 1H),
7.44-7.41 (m, 3H), 7.27-7.21 (m, 3H), 6.91-6.89 (m, 1H), 6.78-6.75
(m, 1H), 3.96 (s, 3H), 3.34-3.28 (m, 4H), 2.55-2.50 (m, 4H), 2.45
(s, 3H), 2.34 (s, 3H), 0.99-0.97 (m, 6H)
Example 56
Production of
(Z)--N-(2-(diethylamino)ethyl)-2,4-dimethyl-5-((2-oxo-5-(5-phenylfuran-2--
yl)indolin-3-ylidene)methyl)-1H-pyrrole-3-carboxamide
##STR00153##
[0400] To a solution of No (31 mg, 0.068 mmol) in DMF/H.sub.2O
(0.75 ml/0.25 ml) was added Pd(PPh.sub.3).sub.4 (15.9 mg, 0.014
mmol), 5-phenylfuran-2-boronic acid (17 mg, 0.090 mmol) and
potassium carbonate (14 mg, 0.100 mmol). The mixture was stirred at
120.degree. C. for 1 hour under microwave irradiation. The mixture
was concentrated in vacuo. The residue was purified by reverse
phase column chromatography (H.sub.2O/CH.sub.3CN) to give
(Z)--N-(2-(diethylamino)ethyl)-2,4-dimethyl-5-((2-oxo-5-(5-phenylfuran-2--
yl)indolin-3-ylidene)methyl)-1H-pyrrole-3-carboxamide (12 mg) as an
orange solid.
[0401] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.76-7.72 (m, 3H),
7.55 (d, 1H, J=9.9 Hz), 7.45-7.37 (m, 3H), 7.27-7.24 (m, 3H), 6.91
(d, 1H, J=8.4 Hz), 6.72 (d, 1H, J=3.3 Hz), 6.67 (d, 1H, J=3.6 Hz),
3.57 (brs, 2H), 2.71 (brs, 4H), 2.58 (s, 3H), 2.52 (s, 3H), 1.11
(brs, 6H).
Test Example 1
Biological Assays
[0402] Compounds of the present invention can be tested according
to the protocol described.
[0403] Cell Culture: FaDu (ATCC, Manassas, Va.) was maintained in
Dulbecco's Modified Eagle Medium (DMEM) (Invitrogen, Carlsbad,
Calif.) supplemented with 10% fetal bovine serum (FBS) (Gemini
Bio-Products, West Sacramento, Calif.) and 5%
penicillin/streptomycin/amphotercin B (Invitrogen).
[0404] Cell Viability Determination: For colony formation assay,
cells were plated in 6 well plates at 2000 cells per well.
Twenty-four hours after plating, cells were treated with compound.
Colonies were allowed to develop for 7-10 days, at which they were
stained with modified Giemsa stain (Sigma). Stained colonies were
then counted to determine ICso.
[0405] Western Blot Analysis: Cultured cells were harvested and
lysed in whole-cell extract buffer (50 mM Tris-HcL pH 7.5, 150 mM
NaCl, 1.0% NP-40, 1 mM EDTA, 0.1 mM sodium orthovanadate, IX
protease inhibitor cocktail (Roche)) by incubation for 30 minutes
on ice. Soluble proteins were separated by centrifugation at
13,000.times.g in a microcentrifuge, and supernatants were stored
at -70.degree. C. Proteins were separated by sodium dodecyl
sulfate-polyacrylamide gel electrophoresis analysis and transferred
to a polyvinylidene difluoride membrane (Biorad, Hercules, Calif.)
by electroblotting.
[0406] CSC isolation with surface markers: Sorting tumor cells
based primarily upon the differential expression of the surface
marker(s), such as CD44 or CD133, have accounted for the majority
of the highly tumorigenic CSCs described to date. CD44.sup.high
cells were isolated by FACS according to the methods described in
Ponti et al, with slight modification [Ponti, D., et al, Isolation
and in vitro propagation of tumorigenic breast cancer cells with
stem/progenitor cell properties. Cancer Res, 2005. 65(13); p.
5506-11.], Briefly, after trypsinization and recovery of cells for
30 minutes at 37.degree. C. in growth media, cells were pelleted at
400.times.g and were resuspended in PBS with 2% FBS and ImM EDTA at
1.times.10.sup.6 cells/mL. Cells were then incubated on ice with a
1:100 dilution of CD44-FITC (BD Biosicences, San Diego, Calif.) for
15 minutes. Alternatively, CD24-PE (BD Bioscences, San Diego,
Calif.) (1:100) was utilized for negative selection.
[0407] After washing three times, cells were resuspended at
2.times.10 VmL and passed through a 40 .mu.M mesh before
sorting.
[0408] Sphere assay: A reliable method of measuring the
self-renewal capacity of cell population if the ability to be
cultured as spheres in the absence of serum or attachment.
CD44.sup.high FaDu cancer stem cells were cultured in ultra low
attachment plates in cancer stem cell media (DMEM/F12, B27
Neurobasal supplement, 20 ng/ml EGF, 10 ng/rnl FGF, 4 .mu.g/ml
insulin, and 0.4% BSA) to allow spheres formation. Typically,
sphere formation was evaluated by microscopy after 10-14 days in
culture and spheres with >50 cells were scored.
TABLE-US-00006 TABLE 5 colonyformation sphere Example IC.sub.50
(.mu.M) IC.sub.50 (.mu.M) 4 6.3 0.6 6 3.6 0.3 7 1.8 0.6 9 3.8 1.3
10 5.4 3 12 24 >50 30 >100 8.9 31 3.7 0.2 32 0.5 0.05 33 2.5
1.7 38 6.2 2 42 1.9 1.3 43 6.5 1.9 45 22 >50 47 6.2 9.1
[0409] As shown in Table 5, compounds of present invention show a
potent inhibitory acidity for sphere formation of CSC.
Test Example 2
Solubility
[0410] Compounds of the present invention can be tested according
to the protocol described.
[0411] A solution of compounds (10 mM in dimethylsulfoxide, 2
.mu.L) was diluted with 50% acetonitrile (198 .mu.L), and this
solution was used as a standard solution. On the other hand, a
solution of compounds (10 mM in dimethylsulfoxide, 15 .mu.L) was
transferred to a 96 well plate, then solvent was removed in vaccuo.
Dimethyl sulfoxide (3 .mu.L) was added to dissolve a residual
solid, and then buffer solution (pH7.4 or 1.2, 300 .mu.L) was
added. The mixture was vigorously stirred for 90 min, then leave it
for 16-20 h. An insoluble material was removed, then the solution
was analyzed by HPLC. The concentration of compound, which was
corresponding to a solubility of the compound, was determined by
the comparison with area value between sample and the standard
solution.
TABLE-US-00007 TABLE 6 pH 1.2 pH 7.4 Example (mg/mL) (mg/ml) 12
>0.2000 >0.2000 26 >0.2000 <0.0005 41 0.186 0.0003 42
>0.2000 0.011 43 0.163 0.0004 48 0.147 <0.0003 49 0.109 0.013
50 0.154 0.007 51 >0.2000 <0.0005 52 0.163 0.004 53
>0.2000 0.061 55 0.043 0.0005
INDUSTRIAL APPLICABILITY
[0412] A compound of formula (1), or a pharmaceutically acceptable
salt thereof is useful a CSCPKs inhibitor to inhibit, reduce or
diminish cancer stem cell survival and/or proliferation in a
mammal. A compound of formula (1), or a pharmaceutically acceptable
salt thereof is also useful an anti cancer agent.
* * * * *