Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
CA 02866502 2016-09-26
Thiochromeno[2,3-c]quinolin-12-one derivatives, preparation
method and application thereof
BACKGROUND OF THE INVENTION
1. FIELD OF THE INVENTION
The invention relates to development of cancer drug, especially
relates to the development of novel
thiochromeno[2,3-c]quinolin-12-one derivatives, preparation method
and application thereof.
2. DESCRIPTION OF THE PRIOR ART
Telomerase is the enzyme that synthesizes telomeric DNA, the
terminal DNA at chromosome ends which, together with
telomere-binding proteins, confers stability to chromosomes. In most
of organism, the replication and maintenance of the length of
telomere has to rely on telomerase. The telomerase is composed of
RNA and protein subunits. At present, part of important telomerase
subunits had been identified. The composition of human telomerase
comprising: human telomerase reverse transcriptase (hTERT) having
reverse transciptase activity, human telomerase RNA component
used as a template, and some telomere-binding proteins such as
human telomerase-associated protein, p23, hsp90, hsp40, hsp70 and
the like.
Many research studies had indicated that the activity of human
telomerase can only be detected in cells having high proliferation
ability, for example, germ cells, hemopoietic cells, part of stem cells,
CA 02866502 2014-10-03
most of immortalized cells and most of tumor cells. In the somatic
cell, the telomere will be shorten gradually as the number of cell
division increased, which may be considered as the mitotic clock for
counting the number of cell division. When a telomere is shortened
to a certain extent, cell will stop division and entering aging stage,
stay at this stage for a period of time, and then goes to death. This
period of time is called mortality stage 1 (MI stage). When a tumor
suppressor gene such as p53 or Rb is mutated within 1\41 stage, the
cell might escape from aging stage and keeps on cell division in this
period of time which is called mortality stage 2 (M2 stage). If a cell
lacks of telomerase activity during this period, the length of a
telomere will be reduced still, the telomere will not be able to protect
the the terminal end of the chromosome, and this might result into the
instability of the chromosome, as well as the cell can not transfer
genetic information completely and enters apoptosis in the end.
Therefore, M2 stage is also called a crisis satge. Most of cells will die
in M2 stage, except small part of cells with telomerase activity will
survive. This small part of cells will continue to divide without
limitation and become an immortalized cell (or a cancer cell).
In view of the foregoing, it is thought generally that the
activation of telomerase can maintain the length of a telomere so as to
prevent a cell from entering the ageing stage; or the inhibition of
telomerase activity can be used to limit the division of a cancer cell.
Both thought may become the key factors in the development of a cell
toward immortalization or cancerization. In summary, using the
telomerase inhibitors to treat the cancer have been considered as a
2
CA 02866502 2014-10-03
, ,
novel cancer-specific therapy, as most tumor cells have high
expression of telomerase, whereas most normal somatic cells express
low or undetectable levels of telomerase and is therefore an attractive
target for the design of anticancer agents.
Cancers arise from abnormal proliferation of DNA. Therefore,
selectively destroy the DNA of cancer cells without damaging the
DNA of normal cells is highly desired. However, it is difficult to
differentiate the DNAs between normal cells and cancer cells.
Consequently, specific 'targeted therapy' was developed following
identification of the differences between normal cells and cancer cells,
and when combined with other chemotherapies or radiation therapies,
targeted therapy can significantly reduce side effects and provide
better treatment outcomes. Thus, targeted therapy currently is a
popular field in studying cancer treatments. Because topoisomerases
have been found to play an indispensible role in DNA replication, they
have become the objects of targeted therapy for anticancer treatments.
The anticancer drug camptothecin discovered by M.E. Wall and M.C.
Wani in 1966 through systematic screening of natural substances is an
inhibitor for type I topoisomerases.
Unfortunately, camptothecin has numerous disadvantages and
thus cannot be used for clinical treatment. For example, the lactone
ring can be easily hydrolyzed to hydroxycarboxylate in vivo at the
normal pH and then binds to serum albumin and lose its effect of
inhibiting the function of type I topoisomerases. In addition, the
structure of the tricomplex of camptothecin-Top I-DNA is not stable
3
CA 02866502 2014-10-03
because the complex is not maintained by covalent bonds and water
solubility of camptothecin is poor which causes lower bioavailability.
The p-glycoprotein (MDR1, ABCB1) efflux transporter proteins in the
cell membrane transported the drugs out of the cells and more
important is that some tumor cells have slowly developed resistance
and adverse drug side effects against camptothecin. As a result, a
number of water-soluble semi-synthetic drugs were developed even
after commercialization of camptothecin such as Topotecan
(HYCAMTIN ) which is used for treating ovarian cancer and
Irinotecan (CAMPTO ) which is used for treating colon cancer and
both have issues when used for clinical treatment.
Hence, based on the importance of topoisomerase inhibitors in
development of anticancer drugs, the inventor of this application
developed a series of novel thiochromeno[2,3 -e]quinolin-12-one
derivatives and disclosed the preparation methods as well as relevant
applications herein after a number of innovative improvements.
SUMMARY OF THE INVENTION
In one aspect, present invention provides a compound as shown
in formulation (I):
0O
CI =s N
(I)
wherein the R is selected from the groups consisting of:
i) halo, amino, hydroxyl and thiol groups;
4
CA 02866502 2014-10-03
. ,
ii) linear alkyl chains of N(CH2)õfl, alkyl groups with
substituted
side chains, alkyl side chains with a substituted amino group
and alkyl side chains with a substituted hydroxyl group,
wherein 1 < n < 10;
iii) 0(CH2)H, N(CH3)2, NH(CH2)nNH(CH2)õOH, wherein 1 5_ n
< 10;
iv) nitrogen-containing cycloalkyl groups and heterocyclic
compounds of C3_12 which contain 1 to 3 heteroatoms selected
from 0, S and N, wherein the ortho-, para- and meta- position
can be further selected independently from one of the groups
consisting of: hydrogen group, (CH2)n alkyl groups, (CH2)n
hydroxyl groups, (CH2)nC3.12 cycloalkyl groups, (CH2)C3-12
nitrogen-containing cycloalkyl groups, (CH2)n benzene rings,
formyl group and (CH2)nC0C3.12 nitrogen-containing
cycloalkyl groups, wherein 0 < n < 10;
v) NH(CH2)nR1, 0 < n < 10, wherein RI is selected from the
groups consisting of: N(CH3)2, C(NH2)2, linear alkyl chains
of NH(CH2)nH, alkyl groups with substituted side chains,
alkyl side chains with a substituted amino group and alkyl
side chains with a substituted hydroxyl group;
vi) NH(CH2)nR2, 0 < n < 10, wherein R2 is selected from the
groups consisting of: benzene rings, C3_12 cycloalkyl groups
and heterocyclic groups of which contain 1 to 3 heteroatoms
selected from 0, S and N, wherein the ortho-, para- and meta-
position can be further selected independently from one of
the groups consisting of: Methoxyl group, amino group,
benzene rings, alkyl, amino, nitro,hydroxyl groups with
5
CA 02866502 2014-10-03
. .
substituted C1-C3 side chains and C3-12 heterocyclic groups;
wherein the C3-12 heterocyclic groups which contain 1 to 3
heteroatoms selected from 0, S and N;
and their pharmaceutically acceptable salts, stereoisomers and
enantimoers.
According to the invention, wherein the R group consisting of i)
¨ vi) are selected from the group consisting of chlorine, hydroxyl,
methoxyl, dimethylamino, piperazin-l-yl, 4-methylpiperazin-1-y1,
4-ethylpiperazin- 1 -yl, 4-(2-hydroxyethyl)piperazin- 1 -yl,
4-Benzylpiperazin-1-y1, 4-phenylpiperazin-1-y1, morpholino,
thiomorpholino, piperidin-l-yl, 4-hydroxypiperidin-l-yl,
4-Benzylpiperidin-1-y1, (1,4'-Bipiperidin)-1'-yl,
4-(3-(piperidin-4-yl)propyl)piperidin- 1 -yl, pyrrolidin- 1 -yl,
2-oxopiperidin-1-y1, methylamino, ethylamino, propylamino,
butylamino, isobutylamino, pentan-3-ylamino,
(2-(dimethylamino)ethyl)amino, (2-(diethylamino)ethyl)amino,
2-ethanolamino, 3-propanolamino, 5-pentanolamino,
(1 -hydroxybutan-2-yl)amino, (4-methylpentan-2-yl)amino,
(2-Aminoethy1)amino, (2-((2-hydroxyethyl)amino)ethyl)amino,
(2-morpholinoethyl)amino, (3-(dimethylamino)propyl)amino,
(3-(diethylamino)propyl)amino,
(34(2-hydroxyethypamino)propyl)amino,
(2,3-dihydro-11-1-inden-2-yDamino, cyclohexylamino,
(1-Benzylpiperidin-4-yl)amino, (thiophen-2-ylmethypamino,
(cyclohexylmethyl)amino, benzylamino, (pyridin-2-ylmethyl)amino,
(Benzo[d][1,3]dioxo1-5-ylmethyl)amino, (2-methoxybenzyl)amino,
(3,4-dimethoxybenzyl)amino, phenethylamino,
6
CA 02866502 2014-10-03
(4-methoxyphenethyl)amino, (4-aminophenethyl)amino, guanidine
and piperidin- 1 -ylamino.
According to the invention, wherein the compound is selected
from the group consisting of:
3-((4-Chlorophenyl)thio)-2-hydroxyquinoline-4-carboxylic acid,
6,9-Dichloro- 1 2H-thiochromeno[2,3 -c]quinolin-1 2-one,
1 0-Chloro-6-hydroxy- 1 2H-thiochromeno [2,3 -c]quinolin- 1 2-one,
1 0-Chloro-6-methoxy- 1 2H-thiochromeno [2,3 -c]quinolin- 12-one 1 O-C
hloro-6-dimethy lam ino- 1 2H-thiochromeno [2,3 -c]quinolin- 12-one,
1 0-Chloro-6-(piperazin- 1 -y1)- 1 2H-thiochromeno [2,3 -c]quinolin- 1 2-on
e,
1 0-Chloro-6-(4-m ethylp ip erazin- 1 -y1)- 1 2H-thiochromeno [2,3 -c]quino
lin- 12-one,
1 0-Chloro-6-(4-ethylpiperazin- 1 -y1)- 1 2H-thiochromeno [2,3 -c]quinoli
n-12-one,
1 0-Chloro-6-(4-(2-hydroxyethyppiperazin- 1 -y1)- 1 2H-thiochromeno [2,
3 -c] quinolin-1 2one,
6-(4 -B enzylp iperazin- 1 -y1)- 1 0-chloro- 1 2H-thiochrom eno [2,3 -c]quinol
in- 12-one,
1 0-Chloro-6-(4-pheny lp iperazin- 1 -y1)- 1 2H-thio chromeno [2,3 -c] quino
lin- 12-one,
1 0-Chloro-6-morpholino-12H-thiochromeno [2,3 -c]quinolin- 12-one,
1 0-Chloro-6-thiomorpholino- 1 2H-thiochromeno [2,3 -c]quinolin- 12-on
e,
1 0-Chloro-6-(piperidin- 1 -y1)- 1 2H-thiochromeno [2,3 -c]quinolin- 12-on
e,
1 0-Chloro-6-(4-hy droxyp iperidin- 1 -y1)- 1 2H-thiochromeno [2,3 -c]quin
7
CA 02866502 2014-10-03
olin-12-one,
6-(4-Benzylpiperidin-l-y1)-10-chloro-12H-thiochromeno[2,3-c]quinol
in-12-one,
6-([1,4'-Bipiperidin]-1'-y1)-10-chloro-12H-thiochromeno[2,3-c]quinoli
n-12-one,
10-Chloro-6-(4-(3-(piperidin-4-yl)propyl)piperidin-1-y1)-12H-thiochr
omeno [2,3 -c]quinolin-12-one,
10-Chloro-6-(pyrrolidin-1-y1)-12H-thiochromeno[2,3-c]quinolin-12-o
ne,
10-Chloro-6-(2-oxopiperidin-l-y1)-12H-thiochromeno[2,3-c]quinolin-
12-one,
10-Chloro-6-methylamino-12H-thiochromeno [2,3 -c]quinolin-12-one,
10-Chloro-6-ethylamino-12H-thiochromeno[2,3-c]quinolin-12-one,
10-Chloro-6-propylamino-12H-thiochromeno[2,3-c]quinolin-12-one,
6-(Butylamino)-10-chloro-12H-thiochromeno [2,3-c]quinolin-12-one,
10-Chloro-6-isobutylamino-12H-thiochromeno [2,3 -c]quinolin-12-one,
10-Chloro-6-(pentan-3-y lam ino)-12H-thiochromeno [2,3-c]quino lin-1
2-one,
10-Chloro-642-(dimethy lam ino)ethy Damino)-12H-thiochromeno [2,3
-c]quinolin-12-one,
10-Chloro-6-02-(diethy lam ino)ethy Dam ino)-12H-thiochromeno [2,3-c
] quinolin-12-one,
10-Chloro-6-(2-ethanolamino)-12H-thiochromeno [2,3-c]quinolin-12-
one,
10-Chloro-6-(3-propanolamino)-12H-thiochromeno[2,3-c]quinolin-12
-one,
10-Chloro-6-(5-pentanolamino)-12H-thiochromeno[2,3-c]quinolin-12
8
CA 02866502 2014-10-03
. ,
. ,
-one,
10-Chloro-6-((1-hydroxybutan-2-yl)amino)-12H-thiochromeno[2,3-c]
quinolin-12-one,
10-Chloro-644-methylpentan-2-yl)amino)-12H-thiochromeno[2,3-c]
quinolin-12-one,
6-((2-Aminoethy1)amino)-10-chloro-12H-thiochromeno[2,3-c]quinoli
n-12-one,
10-Chloro-64242-hydroxyethypamino)ethypamino)-12H-thiochro
meno[2,3-c]quinolin-12-one,
10-Chloro-6-((2-morpholinoethyl)amino)-12H-thiochromeno[2,3-c]qu
inolin-12-one,
10-Chloro-6-((3-(dimethylamino)propyl)amino)-12H-thiochromeno[2,
3-c]quinolin-12-one,
10-Chloro-6-((3-(diethylamino)propyl)amino)-12H-thiochromeno[2,3
-c]quinolin-12-one,
10-Chloro-64342-hydroxyethypamino)propypamino)-12H-thiochro
meno[2,3-clquinolin-12-one,
10-Chloro-6-((2,3-dihydro-1H-inden-2-yl)amino)-12H-thiochromeno[
2,3-c]quinolin-12-one,
10-Chloro-6-(cyclohexylamino)-12H-thiochromeno[2,3-c]quinolin-12
-one,
6-((1-Benzylpiperidin-4-yl)amino)-10-chloro-12H-thiochromeno[2,3-
c]quinolin-12-one,
10-Chloro-6-((thiophen-2-ylmethyDamino)-12H-thiochromeno[2,3-c]
quinolin-12-one,
10-Chloro-6-((cyclohexylmethyl)amino)-12H-thiochromeno[2,3-c]qui
nolin-12-one,
9
CA 02866502 2014-10-03
. .
. .
6-(B enzylam ino)- 1 0-chloro- 1 2H-thio chromeno [2,3 -c] quinolin- 12-one
,
1 0-Chloro-6-((pyri din-2-y lmethy 1)am ino)- 1 2H-thiochromeno [2,3 -c]q
uinolin- 12-one,
6-((Benzo [d] [1 ,3 ]dioxo1-5 -ylmethyl)amino)- 1 0-chloro- 1 2H-thiochrom
eno [2,3 -c]quinolin- 12-one,
1 0-Chloro-6-((2-m ethoxy benzy 1)am ino)- 1 2H-thiochrom eno [2,3 -c] qui
nolin- 12-one,
1 0-Chloro-6((3,4-dimethoxybenzypamino)-12H-thiochromeno [2,3-c]
1 0 quinolin- 12-one,
1 0-Chloro-6-(phenethylamino)- 1 2H-thio chrom eno [2,3 -c] quinolin- 1 2-
one,
1 0-Chloro-6-((4-methoxyphenethy Dam ino)- 1 2H-thiochrom eno [2,3 -c]
quinolin- 12-one,
6-((4-Am inophenethyDamino)- 1 0-chloro- 1 2H-thiochromeno [2,3 -c] qu
inolin- 12-one,
2-( 1 O-Chloro- 1 2-oxo- 1 2H-thiochromeno [2 ,3 -c]quinolin-6-yl)guanidin
e,
1 0-Chloro-6-(piperidin- 1 -y lam ino)- 1 2H-thio chrom eno [2,3 -c] quinolin-
12-one,
and their salts.
In another aspect, the invention provides a pharmaceutical
composition comprising an effective amount of the abovementioned
compound and at least one pharmaceutically acceptable vehicle,
diluent or excipient.
In another aspect, the invention provides a method for inhibiting
Topoisomerase I activity which comprises administrating an effective
CA 02866502 2014-10-03
amount of the abovementioned compound.
In another aspect, the invention provides a method for inhibiting
Topoisomerase II activity which comprises administrating an effevtive
amount of the compound according to claim 1.
In another aspect, the invention provides a method for the
treatment of cancer which comprises administrating an effective
amount of the compound according to claim 1.
According to the inventionõ wherein the cancer is selected from
the groups consisting of leukemia, non-small cell lung cancer,
colorectal cancer, central nervous system (CNS) cancer, melanoma,
ovarian cancer, renal cancer, prostate cancer and breast cancer.
In another aspect, the invention provides a method for
preparation of thiochromeno[2,3-c]quinolin-12-one derivatives,
wherein the method comprising:
(1) mix isatin, 2-((4-chlorophenyl)thio)acetic acid and sodium
acetate was heated at 150 C for 1 h, after cooling the mixture
was added acetic acid, the precipitate was collected, washed
with acetic acid, water and n-hexane, and obtained compound
2 (3-((4-Chlorophenyl)thio)-2-hydroxyquinoline-4-carboxylic
acid);
(2) a solution of compound 2 (3-((4-Chlorophenyl)thio)-
2-hydroxyquinoline-4-carboxylic acid) in phosphoryl
trichloride was heated at 150 C for 48 h, after cooling the
mixture was poured into water 0 C, the precipitate was
collected by filtration, then added into 10% NaHCO3 with
vigorous stirring for 1 h, the resulting precipitate was
collected and washed with H20, the crude solid was
11
CA 02866502 2014-10-03
, .
. .
recrystallized by dichloromethane to give compound 3
(6,9-Dichloro-12H-thiochromeno[2,3-c] quinolin-12-one);
(3) a solution of compound 3 (6,9-Dichloro-12H-thiochromeno
[2,3-c]quinolin-12-one) in DMF was added conc. HC1 and
refluxed, after 6 hours, the conc. HC1 was added dropwise
and refluxed for another 12 hours, the mixture was
evaporated in vacuo and treated with H20, after filtered the
crude solid was washed with Et0H to give compound 4
(10-Chloro-6-hydroxy-12H-thiochromeno[2,3-c]quinolin-12-
one);
(4) a suspension of compound 3 (6,9-Dichloro-12H-
thiochromeno[2,3-c]quinolin-12-one) and sodium methoxide
in methanol was refluxed for 16 h, after cooled the solvent
was removed, filtrated and washed with ethanol and n-hexane
to collect compound 5 (10-Chloro-6-methoxy-12H-
thiochromeno[2,3-c]quinolin-12-one);
(5) a solution of compound 3 (6,9-Dichloro-12H-
thiochromeno[2,3-c]quinolin-12-one), appropriate secondary
amines and sodium carbonate in DMSO was refluxed for 10
hours, then the reaction was added ice-water, the precipitate
was filtered, washed with water/methanol and collected to get
compound 6-21:
10-Chloro-6-dimethylamino-12H-thiochromeno[2,3-c]quinoli
n-12-one,
10-Chloro-6-(piperazin-1-y1)-12H-thiochromeno[2,3-c]quinol
in-12-one,
10-Chloro-6-(4-methylpiperazin-1-y1)-12H-thiochromeno[2,3
12
CA 02866502 2014-10-03
. .
. .
-c]quinolin-12-one,
10-Chloro-6-(4-ethylpiperazin-1 -y1)-12H-thiochromeno [2,3 -c
]quinolin-12-one,
10-Chloro-6-(4-(2-hydroxyethyl)piperazin-1-y1)-12H-thiochr
omeno[2,3 -c]quinolin- 12one,
6-(4-Benzylpiperazin-1-y1)-10-chloro-12H-thiochromeno [2,3 -
clquinolin-12-one,
10-Chloro-6-(4-pheny lp iperazin-1 -y1)-12H-thiochromeno [2 ,3
-c] quino lin-12-one,
10-Chloro-6-morpholino-12H-thiochromeno [2,3 -c]quinolin-1
2-one,
10-Chloro-6-thiomorpholino-12H-thiochromeno [2,3 -c]quinol
in-12-one,
10-Chloro-6-(piperidin-1-y1)-12H-thiochromeno [2,3-c] quinol
in-12-one,
10-Chloro-6-(4-hydroxypiperidin-1-y1)-12H-thiochromeno [2,
3 -c]quinolin-12-one,
6-(4 -Benzylpiperidin-l-y1)-10-chloro-12H-thiochromeno [2,3-
c] quinolin-12-one,
6-([1,4'-Bipiperidin]-1'-y1)-10-chloro-12H-thiochromeno[2,3 -
c] quinolin-12-one,
10-Chloro-6-(4-(3-(piperidin-4-yl)propyl)piperidin-1-y1)-12H
-thiochromeno[2,3-c]quinolin-12-one,
10-Chloro-6-(pyrrolidin-1-y1)-12H-thiochromeno [2,3 -c]quino
lin-12-one, and
10-Chloro-6-(2-oxopip eridin-1 -y1)-12H-thio chrom eno [2,3-c]
quinolin-12-one respectively;
13
CA 02866502 2014-10-03
(6) a solution of compound 3 (6,9-Dichloro-12H-
thiochromeno[2,3-c]quinolin-12-one) in DMSO was added
appropriate primary amines and refluxed for 8 hours, after
cooled the reaction was added water, the precipitate was
filtered and washed with water and methanol to collect
compound N1¨N34:
10-Chloro-6-methylamino-12H-thiochromeno[2,3-c]quinolin-
12-one,
10-Chloro-6-ethylamino-12H-thiochromeno[2,3-c]quinolin-1
2-one,
10-Chloro-6-propylamino-12H-thiochromeno[2,3-c]quinolin-
12-one,
6-(Butylamino)-10-chloro-12H-thiochromeno[2,3-c]quinolin-
12-one,
10-Chloro-6-isobutylamino-12H-thiochromeno[2,3-c]quinoli
n-12-one,
10-Chloro-6-(pentan-3-ylamino)-12H-thiochromeno[2,3-c]qu
inolin-12-one,
10-Chloro-642-(dimethylamino)ethy1)amino)-12H-thiochro
meno[2,3-c]quinolin-12-one,
10-Chloro-64(2-(diethylamino)ethypamino)-1211-thiochrome
no[2,3-c]quinolin-12-one,
10-Chloro-6-(2-ethanolamino)-12H-thiochromeno[2,3-c]quin
olin-12-one,
10-Chloro-6-(3-propanolamino)-12H-thiochromeno[2,3-c]qui
nolin-12-one,
10-Chloro-6-(5-pentanolamino)-12H-thiochromeno[2,3-c]qui
14
CA 02866502 2014-10-03
, A
. .
nolin-12-one,
10-Chloro-64(1-hydroxybutan-2-yl)amino)-12H-thiochromen
o[2,3-c]quinolin-12-one,
10-Chloro-6-((4-methylpentan-2-yDamino)-12H-thiochromen
o[2,3-c]quinolin-12-one,
6-((2-Aminoethyl)amino)-10-chloro-12H-thiochromeno[2,3-c
]quinolin-12-one,
10-Chloro-642-((2-hydroxyethyl)amino)ethypamino)-12H-t
hiochromeno[2,3-c]quinolin-12-one,
10-Chloro-642-morpholinoethyl)amino)-12H-thiochromeno[
2,3-c]quinolin-12-one,
10-Chloro-643-(dimethylamino)propypamino)-12H-thiochr
omeno[2,3-c]quinolin-12-one,
10-Chloro-643-(diethylamino)propyl)amino)-12H-thiochro
meno[2,3-c]quinolin-12-one,
10-Chloro-6-43-((2-hydroxyethyDamino)propyl)amino)-12H
-thiochromeno[2,3-c]quinolin-12-one,
10-Chloro-6-((2,3-dihydro-1H-inden-2-y1)amino)-12H-thioch
romeno[2,3-c]quinolin-12-one,
10-Chloro-6-(cyclohexylamino)-12H-thiochromeno[2,3-c]qui
nolin-12-one,
6-((1-Benzylpiperidin-4-yl)amino)-10-chloro-12H-thiochrom
eno[2,3-c]quinolin-12-one,
10-Chloro-6-((thiophen-2-ylmethypamino)-12H-thiochromen
o[2,3-c]quinolin-12-one,
10-Chloro-6-((cyclohexylmethypamino)-12H-thiochromeno[
2,3-c]quinolin-12-one,
CA 02866502 2014-10-03
) f
. .
6-(Benzylamino)-10-chloro-12H-thiochromeno[2,3-c]quinoli
n-12-one,
10-Chloro-6-((pyridin-2-ylmethyl)amino)-12H-thiochromeno
[2,3-c]quinolin-12-one,
6-((Benzo[d][1,3]dioxo1-5-ylmethypamino)-10-chloro-12H-t
hiochromeno[2,3-c]quinolin-12-one,
10-Chloro-6-((2-methoxybenzyl)amino)-12H-thiochromeno[2
,3-c]quinolin-12-one,
10-Chloro-6-((3,4-dimethoxybenzyl)amino)-12H-thiochrome
no[2,3-c]quinolin-12-one,
10-Chloro-6-(phenethylamino)-12H-thiochromeno[2,3-c]quin
olin-12-one,
10-Chloro-644-methoxyphenethyDamino)-12H-thiochromen
o[2,3-c]quinolin-12-one,
6-((4-Aminophenethyl)amino)-10-chloro-12H-thiochromeno[
2,3-c]quinolin-12-one,
2-( 1 O-Chloro- 1 2-oxo- 1 2H-thiochrom eno [2,3 -c] quino lin-6-y1)
guanidine, and
1 O-Chloro-6-(p iperidin- 1 -y larn ino)- 1 2H-thiochromeno [2,3 -c]
quinolin-12-one respectively.
These features and advantages of the present invention will be
fully understood and appreciated from the following detailed
description of the accompanying Drawings.
BRIEF DESCRIPTION OF THE DRAWINGS
Fig. 1 depicts the general scheme for a series of thiochromeno
[2,3 -c] quino lin- 12-one derivatives.
16
CA 02866502 2014-10-03
=
Fig. 2 is the effect of compounds 5, 7, 8, 16, 19 and CPT on
DNA relaxation catalyzed by TOP I at a concentration of 25 ,M and
50 M.
Fig. 3 is the effect of compounds N2, N7, N8, N9, N14-N19,
N25, and CPT on DNA relaxation catalyzed by TOP I at a
concentration of 50 M.
Fig. 4a-d are the effects of compounds 7, N7, N14, N15, N18,
N19 and N25 on TOP I mediated supercoiled DNA relaxation.
Fig. 5 is the effect of compounds 5, 7, 8, 16, 19 and VP-16 on
DNA relaxation catalyzed by TOP II at a concentration of 25 1A,M and
50 M.
Fig. 6 is the effect of compounds N2, N7, N8, N9, N14-N19,
N25, and VP-16 on DNA relaxation catalyzed by TOP II at a
concentration of 501.1M.
Fig. 7a-d are the effects of compounds 7, N7, N8, N14, N15,
N18, and N19 on TOP II mediated supercoiled DNA relaxation.
DETAILED DESCRIPTION OF THE PREFERRED
EMBODIMENT
Unless defined otherwise, all technical and scientific terms used
herein have the meaning commonly understood by a person skilled in
the art to which this invention belongs. As used herein, the following
terms have the meanings ascribed to them unless specified otherwise.
The present invention will now be described more specifically with
reference to the following embodiments, which are provided for the
17
CA 02866502 2014-10-03
purpose of demonstration rather than limitation.
The term "treatment", "under treatment" and similar terms
refer to the methods which ameliorate, improve, reduce or reverse the
patient's disease or any relevant symptoms caused by the disease, or
methods which can prevent onset of such diseases or any resulting
symptoms.
The term "pharmaceutically acceptable" is used to describe
substances to be used in the composition must be compatible with
other ingredients in the formulation and be harmless to the subject.
The inventive composition can be prepared into a dosage form
for suitable application of the inventive composition by using
technology commonly understood by a person skilled in the art
through formulating the abovementioned Lactobacillus isolated
strain(s) with a pharmaceutically acceptable vehicle, wherein the
excipients include, but are not limited to, solution, emulsion,
suspension, powder, tablet, pill, lozenge, troche, chewing gum, slurry,
and other suitable forms.
The pharmaceutically acceptable vehicle may contain one or
several reagents selecting form the following list: solvents, emulsifiers,
suspending agents, decomposers, binding agents, excipients,
stabilizing agents, chelating agents, diluents, gelling agents,
preservatives, lubricants, surfactants and other agents suitable for use
in the invention.
In the abovementioned compositions, one or more dissolving aids,
buffers, preservatives, colorants, fragrances, flavoring agents and the
18
CA 02866502 2014-10-03
like, which are commonly used for formulation can be added as
desired.
The term "pharmaceutically acceptable excipients", as used
herein, refers to substances known by persons skilled in the art, which
are physiologically inert, pharmacologically inactive and are
compatible with the physical as well as chemical characteristics of
sorafenib or GW5074. Pharmaceutically acceptable excipients include,
but are not limited to, polymers, resins, plasticizers, fillers, lubricants,
diluents, binders, disintegrants, solvents, co-solvents, surfactants,
preservatives, sweetening agents, flavoring agents, pharmaceutical
grade dyes or pigments, and viscosity agents.
The term "pharmaceutical composition" is used to describe
solid or liquid compositions in a form, concentration and purity that
are suitable for administration in patients (e.g. humans or animals) and
can induce desired physiological changes following administration.
Pharmaceutical compositions are typically sterile and non-pyrogenic.
The present invention will now be described more specifically
with reference to the following embodiments, which are provided for
the purpose of demonstration rather than limitation. The drugs as well
as biomaterials used in the invention are all commercially available
materials and the sources disclosed below are merely examples.
All reactions were monitored by thin-layer chromatography
(TLC) coated with silica gel 60 F254. Melting points of all synthetic
compounds were measured with Mehl B-545 melting point apparatus.
1H NMR: Varian GEMINI-300 (300 MHz) or Agilent 400 MR DD2
19
CA 02866502 2014-10-03
(400 MHz); 8 values are in ppm relative to tetramethylsilane (TMS) as
an internal standard (0 ppm). Multiplicities are recorded as s (singlet),
d (doublet), t (triplet), q (quartet), quin (quintuplet), sext (sextet), sep
(septet), m (multiplet), dd (doublet of doublet), dt (doublet of triplet),
td (triplet of doublet), qd (quartet of doublet) and br (broadened).
Mass spectra: High resolution electrospray ionization (HRESI):
Finnigan MAT 95S (Instrumentation Center, National Taiwan
University, Taipei, Taiwan). X-ray Single Crystal Diffraction: Bruker
Enraf-Nonius APEX II diffractometer (Department of Chemistry,
National Taiwan Normal University). Typical experiments illustrating
the general procedures for the preparation of the
thiochromenoquinolones are described below (Fig. 1).
General procedures for chemical synthesis
General procedure A Preparation of compound 2
A mixture of isatin (1) (0.44 g, 2.99 mmol),
2-((4-chlorophenyl)thio)acetic acid (0.70 g, 3.47 mmol), and sodium
acetate (0.05 g) was heated at 150 C in miniclave for 1 h (TLC
monitored). After cooling, the mixture was added acetic acid 10 mL,
and the gray precipitate was collected, washed with acetic acid, water
and n-hexane, and obtained light purple compound.
General procedure B Preparation of compound 3
A solution of compound 2 (0.55 g, 2.1 mmol) in phosphoryl
trichloride (5 mL) was heated at 150 C for 48 h. After cooling, the
mixture was poured into ice (50 mL) at 0 C. The resulting green
precipitate that separated was collected by filtration. The filtered cake
CA 02866502 2014-10-03
was suspended in 10% NaHCO3 solution (50 mL) with vigorous
stirring for 1 h. The resulting precipitate was collected and washed
with H20. The crude solid was recrystallized from dichloromethane to
give yellow product.
General procedure C Preparation of compound 4
To a solution of compound 3 (0.32 g, 0.96 mmol) in DMF (20
mL) was added conc. HC1 (3 mL) and refluxed. After 6 h, the conc.
HC1 (6 mL) was added dropwise and refluxed for another 12 h. The
mixture was evaporated in vacuo and treated with H20 (20 mL), after
filtered the crude solid was washed with Et0H to give yellow solid.
General procedure D Preparation of compound 5
A suspension of compound 3 (0.33 g, 1.0 mmol) and sodium
methoxide (0.55g, 10 mmol) in methanol (20 mL) was refluxed for 16
h. After cooled, the solvent was removed by rotarvapor vacuum,
filtrated and washed with ethanol and n-hexane to collect the white
solid.
General procedure E Preparation of compounds 6-21
Compound 3 (0.33 g, 1.0 mmol), appropriate secondary amines
(1.1 mmol) and sodium carbonate (5 mmol) in DMSO (20 mL) was
refluxed for 10 h (TLC monitored). After 30 min, the reaction was
added ice-water (100 mL). The precipitate was filtered, washed with
water/methanol and collected to get the yellow solid.
General procedure F Preparation of compounds N1-N32
To a solution of compound 3 (0.33 g, 1.0 mmol) in DMSO (30
mL) was added appropriate primary amines (1.1 mmol) and refluxed
for 8 h (TLC monitored). After cooled, the reaction was added water
21
CA 02866502 2014-10-03
. .
. ,
(100 mL). The precipitate was filtered and washed with water and hot
methanol to collect the yellow solid.
Example 1
3((4-ChlorophenyOthio)-2-hydroxyquinoline-4-carboxylic acid
5 (TC-SCI) (2)
The pure compound was obtained as a gray solid (yield 86%). (Rf
= 0.5 at EA: AcOH = 20: 1). Mp 306-308 C. 114 NMR (300 MHz,
DMSO-d6): 6 (ppm) 7.26 (3H, t, J= 7.6 Hz, Ar-H), 7.34 (2H, d, J=
6.0 Hz, Ar-H), 7.39 (1H, d, J= 8.0 Hz, Ar-H), 7.46 (1H, d, J- 8.0 Hz,
10 Ar-H), 7.62 (1H, t, J = 8.0 Hz, Ar-H), 12.22 (1H, s, -COOH). 13C
NMR (100 MHz, DMSO-d6): 6 (ppm) 115.58, 116.26, 120.36, 123.21,
126.21, 129.33, 130.30, 131.47, 132.54, 134.36, 140.11, 151.69,
159.37, 166.80 (CO). HRMS (ESI) calcd for C16HI0NO3SC1 [MT
331.0070; found [M+Hr- 332.0147 (100), [M+H+2r. 334.0122 (33);
15 found [M-H] 330.0002.
Example 2
6,9-Dichloro-12H-thiochromeno[2,3-chuinolin-12-one (3)
The yellow solid material was isolated in 90% yield (Rf = 0.50 at
CH2C12: n-hexane = 1: 1). Mp 259-261 C (CH2C12). 1H NMR (400
20 MHz, CDC13): 6 (ppm) 7.71 (2H, m, Ar-H), 7.77-7.85 (2H, m, Ar-H),
8.10-8.13 (m, 1H, Ar-H), 8.60 (t, 1H, J= 1.2 Hz, Ar-H), 9.67-9.71 (1H,
m, Ar-H). 13C NMR (100 MHz, CDC13): 6 (ppm) 124.87, 126.28,
127.85, 129.17, 129.39, 129.93, 130.31, 131.20, 131.90, 133.01,
133.09, 133.38, 134.43, 145.27, 146.61, 180.64 (CO). HRMS (ESI)
25 calcd for C16H7NOSC12 [Mr- 330.9625; found [M+Hr 331.9699 (100),
[M+H+2] 333.9672 (67), [M+H+4] 335.9645 (11).
22
CA 02866502 2014-10-03
. =
Example 3
10-Chloro-6-hydroxy-12H-thiochromeno[2,3-clquinolin-12-one
(4)
The yellow solid material was isolated in 95% yield (Rf = 0.40 at
EA). Mp > 410 C. 11-1 NMR (400 MHz, DMSO-d6): 6 ppm 7.35 (1H,
td, J= 7.2, 1.2 Hz, Ar-H), 7.47 (1H, dd, J= 8.4, 1.2 Hz, Ar-H), 7.59
(1H, td, J= 7.2 Hz, 1.6 Hz, Ar-H), 7.89 (1H, dd, J= 8.4 Hz, 2.4 Hz,
Ar-H), 8.10 (1H, d, J= 8.8 Hz, Ar-H), 8.38 (1H, d, J= 2.4 Hz, Ar-H),
9.35 (1H, dd, J= 8.4, 2.4 Hz, Ar-H), 12.73 (br, 1H, -OH). 13C NMR
(75 MHz, CDC13): 6 (ppm) 116.61, 117.52, 123.65, 126.82, 128.44,
130.22, 130.49, 130.54, 132.52, 133.00, 133.42, 135.09, 136.27,
138.90, 158.70, 180.38 (CO). HRMS (ESI) m/z calcd for
Ci6H8NO2SC1 [Mr: 312.9964, found, 314.0051.
Example 4
10-Chloro-6-methoxy-12H-thiochromeno[2,3-clquinolin-12-one
(5)
The gray solid material was isolated in 91% yield (Rf = 0.52 at
CH2C12: n-hexane = 1: 1). Mp 227-228 C. 11-1 NMR (400 MHz,
CDC13): 6 (ppm) 4.27 (3H, s, -OCH3), 7.60 (1H, td, J= 7.6, 1.2 Hz,
Ar-H), 7.37 (1H, d, J= 2.0 Hz, Ar-H), 7.70 (1H, td, J= 7.6 Hz, 1.6 Hz,
Ar-H), 7.94 (1H, dd, J= 8.0 Hz, 1.2 Hz, Ar-H), 8.60 (1H, d, J= 1.6
Hz, Ar-H), 9.64 (1H, dd, J= 8.8 Hz, 1.2 Hz, Ar-H). 13C NMR (100
MHz, CDC13): 6 (ppm) 54.83 (OCH3), 122.91, 126.23, 126.54, 126.76,
127.66, 127.95, 129.23, 129.50, 130.54, 132.49, 133.48, 133.85,
143.82, 156.06, 180.47 (CO). HRMS (ESI) m/z calcd for
C17H10NO2SC1 [Mr 327.0121; found [M+H]+ 328.0203, [M+H+2]+
330.0172.
23
CA 02866502 2014-10-03
,
Example 5
10-Chloro-6-dimethylamino-12H-thiochromeno[2,3-chuinolin-
12-one (6)
Product 6 was prepared from 3 and dimethylamine. The
light-yellow solid material was isolated in 85% yield (Rf = 0.45 at
CH2C12: n-hexane = 1: 1). Mp 194-195 C. 1H NMR (400 MHz,
CDC13): 6 (ppm) 3.06 (6H, s, -CH3), 7.59-7.67 (3H, m, Ar-H), 7.71
(1H, t, J= 7.2 Hz, Ar-H), 8.00 (1H, d, J= 8.4 Hz, Ar-H), 8.59 (1H,d,J
= 1.2 Hz, Ar-H), 9.60 (1H, d, J= 8.4 Hz, Ar-H). 13C NMR (100 MHz,
CDC13): 6 (ppm) 43.00, 123.44, 125.71, 127.22, 127.78 128.47,
129.07, 129.37, 130.59, 130.67, 132.23, 132.46, 133.61, 134.36,
144.84, 158.32, 181.52 (CO). HRMS (ESI) calcd for C181113N20SC1
[M]+ 340.0437; found [M+H] + 341.0517 (100), [M+H+21+ 343.0501
(33).
Example 6
10-Chloro-6-03iperazin-1-y0-12H-thiochromeno[2,3-elquinolin
-12-one (7)
Product 7 was prepared from 3 and piperazine. The dark-yellow
solid material was isolated in 69% yield (Rf = 0.12 at EA: MeOH:
ammonia water = 20: 5: 1). Mp 211-213 C. 1H NMR (400 MHz,
CDC13): 6 (ppm) 3.20 (4H, t, J= 4.8 Hz, -CH2-), 3.36 (4H, t, J= 4.8
Hz, -CH2-), 7.60-7.66 (311, m, Ar-H), 7.70 (1H, td, J = 8.0, 1.2 Hz,
Ar-H), 7.99 (1H, dd, J= 8.4, 0.8 Hz, Ar-H), 8.56 (1H, d, J= 2.0 Hz,
Ar-H), 9.60 (1H, dd, J = 8.4, 0.8 Hz, Ar-H). 13C NMR (100 MHz,
CDC13): 6 (ppm) 45.95, 52.31, 123.62, 125.81, 127.58, 127.86, 128.72,
129.10, 129.37, 130.65, 130.99, 132.17,132.47, 133.63, 134.33,
144.94, 157.61, 181.45 (CO). HRMS (ESI) calcd for C20H16N30SC1
24
CA 02866502 2014-10-03
[M]' 381.0703; found [M+H]' 382.0783.
Example 7
10-Chloro-6-(4-methylpiperazin-1-y1)-12H-thiochromeno[2,3-eku
inolin-12-one (8)
Product 8 was prepared from 3 and 1-methylpiperazine. The
green-yellow solid material was isolated in 80% yield (Rf= 0.24 at EA:
methanol = 5: 1). Mp 212-214 C. 1H NMR (400 MHz, CDC13): 6
(ppm) 2.51 (3H, s, -CH3), 2.84 (4H, br, -CH2-), 3.50 (4H, t, J= 4.5 Hz,
-CH2-), 7.60-7.66 (3H, m, Ar-H), 7.68-7.72 (1H, td, J= 8.1, 1.5 Hz,
Ar-H), 8.01 (1H, dd, J= 8.1, 1.5 Hz, Ar-H), 8.56 (1H, d, J= 1.5 Hz,
Ar-H), 9.60 (1H, dd, J = 8.4, 1.5 Hz, Ar-H). 13C NMR (100 MHz,
CDC13): 6 (ppm) 45.73, 50.31, 54.65, 123.70, 125.90, 127.60, 127.80,
128.82, 129.20, 129.39, 130.62, 130.84, 132.36, 132.47, 133.78,
134.24, 145.08, 157.18, 181.42 (CO). HRMS (ESI) calcd for
C21H18N30SC1 [M]' 395.0859; found [M+H]' 396.0926.
Example 8
10-Chloro-6-(4-ethylpiperazin-1-y0-12H-thiochromeno[2,3-ek
uinolin-12-one (9)
Product 9 was prepared from 3 and 1-ethylpiperazine. The yellow
solid material was isolated in 74% yield (Rf= 0.48 at EA: Me0H = 10:
1). Mp 196-198 C. NMR (400
MHz, CDC13): 6 (ppm) 1.19 (3H, t,
J = 7.2 Hz, -CH3), 2.58 (2H, q, J = 7.2 Hz, -CH2-), 2.78 (4H, br,
-CH2-), 3.46 (4H, t, J = 4.4 Hz, -CH2-), 7.61-7.66 (3H, m, Ar-H),
7.68-7.73 (1H, td, J= 8.4, 1.6 Hz, Ar-H), 8.01 (1H, dd, J= 8.0, 1.2 Hz,
Ar-H), 8.59 (1H, d, J = 4.0 Hz, Ar-H), 9.62 (1H, dd, J = 8.4, 0.8 Hz,
Ar-H). 13C NMR (100 MHz, CDC13): 6 (ppm) 12.07, 50.88, 52.46,
52.67, 123.55, 125.77, 127.48, 127.83, 128.78, 129.12, 129.36, 130.62,
CA 02866502 2014-10-03
, .
. .
130.76, 132.21, 132.47, 133.62, 134.34, 144.97, 157.34, 181.50 (CO).
HRMS (ESI) calcd for C22H20N30SC1 [M]+ 409.1016; found [M+Hr
410.1069.
Example 9
5 10-Chloro-6-(4-(2-hydroxyethyl)piperazin-1-y1)-12H-thiochrom
eno[2,3-c]quinolin-12one (10)
Product 10 was prepared from 3 and 2-(piperazin-1-yl)ethanol.
The green-yellow solid material was isolated in 60% yield (Rf = 0.37
at EA: Me0H= 2: 1). Mp 211-213 C. 1H NMR (400 MHz, CDC13): 5
10 (ppm) 2.74 (2H, t, J= 5.2 Hz, -CH2-), 2.87 (4H, t, J= 3.6 Hz, -CH2-),
3.45 (4H, t, J = 3.6 Hz, -CH2-), 3.72 (2H, t, J = 5.2 Hz, -CH20-),
7.62-7.67 (3H, m, Ar-H), 7.72 (1H, td, J = 7.2, 1.6 Hz, Ar-H), 8.01
(1H, dd, J= 8.4, 1.2 Hz, Ar-H), 8.59 (1H, d, J= 0.6 Hz, Ar-H), 9.62
(1H, dd, J= 4.8, 1.2 Hz, Ar-H). 13C NMR (100 MHz, CDC13): 6 (ppm)
15 50.85, 52.72, 57.74, 59.35, 123.65, 125.81, 127.66, 127.82, 128.73,
129.14, 129.44, 130.68, 130.77, 132.19, 132.51, 133.69, 134.21,
144.90, 157.26, 181.43 (CO). HRMS (ESI) calcd for C22H20N302SC1
[Mr 425.0965; found [M+HT 426.1024.
Example 10
20 6-(4-Benzylpiperazin-1-y1)-10-chloro-12H-thiochromeno[2,3-ch
uinolin-12-one (11)
Product 11 was prepared from 3 and 4-benzylpiperazine. The
yellow solid material was isolated in 81% yield (Rf = 0.43 at EA:
n-hexane = 1: 4). Mp 191-193 C. 1H NMR (400 MHz, CDC13): 5
25 (ppm) 2.78 (4H, br, -CH2N-), 3.43 (4H, t, J= 4.85 Hz, - NCH2-), 3.68
(2H, s, -CH2-), 7.27-7.42 (5H, m, Ar'-H), 7.61-7.67 (3H, m, Ar-H),
7.71 (1H, td, J = 7.6, 1.6 Hz, Ar-H), 8.00 (1H, dd, J = 8.4, 1.2 Hz,
26
CA 02866502 2014-10-03
Ar-H), 8.58 (1H, d, J = 2.0 Hz, Ar-H), 9.61 (1H, dd, J = 8.4, 1.2 Hz,
Ar-H). 13C NMR (100 MHz, CDC13): 6 (ppm) 50.94, 52.96, 63.08,
123.57, 125.78, 127.18, 127.48, 127.83, 128.33, 128.73, 129.09,
129.17, 129.34, 130.58, 130.90, 132.18, 132.44, 133.60, 134.36,
138.11, 144.95, 157.48, 181.47 (CO). HRMS (ESI) calcd for
C27H22N30SC1 [M] 471.1172; found [M+H] 472.1241.
Example 11
10-Chloro-6-(4-phenylpiperazin-1-y0-12H-thiochromeno[2,3-4
quinolin-12-one (12)
Product 12 was prepared from 3 and 1-phenylpiperazine. The
yellow solid material was isolated in 77% yield (Rf = 0.73 at EA:
n-hexane = 1: 4). Mp 236-237 C. 1H NMR (300 MHz, CDC13): 6
(PPm) 3.50-3.60 (8H, m, -CH2-), 6.94 (1H, t, J= 7.2 Hz, Ar'-H), 7.04
(2H, d, J= 8.4 Hz, Ar'-H), 7.33 (2H, t, J= 7.5 Hz, Ar'-H), 7.63-7.67
(3H, m, Ar-H), 7.71 (1H, t, J= 7.2 Hz, Ar-H), 8.02 (1H, d, J = 7.2 Hz,
Ar-H), 8.58 (1H, s, Ar-H), 9.63 (1H, d, J = 8.1 Hz, Ar-H). 13C NMR
(75 MHz, CDC13): 6 (ppm) 48.60, 50.33, 115.77,119.51, 123.21,
125.38, 127.14, 127.30, 128.27, 128.65, 128.71, 128.88, 130.23,
131.77, 131.97, 133.22, 133.70, 144.52, 150.90, 156.83, 159.91,
180.91 (CO). HRMS (ESI) calcd for C26H20N30SC1 [M] 457.1016;
found [M+Hr 458.1095.
Example 12
10-Chloro-6-morpholino-12H-thiochromeno[2,3-4quinolin-12-
one (13)
Product 13 was prepared from 3 and morpholine. The yellow
solid material was isolated in 70% yield (Rf = 0.42 at CH2C12). MP
217-218 C. 1H NMR (300 MHz, CDC13): 6 (ppm) 3.41 (4H, t, J = 4.5
27
CA 02866502 2014-10-03
f
I
Hz, -NCH2-), 4.02 (4H, t, J = 4.5 Hz, -CH20-), 7.62-7.70 (3H, m,
Ar-H), 7.73 (1H, td, J= 7.5, 1.5 Hz, Ar-H), 8.03 (1H, dd, J 8.4, 1.5
Hz, Ar-H), 8.59 (1H, dd, J= 2.1, 0.6 Hz, Ar-H), 9.35 (1H, dd, J= 8.7,
1.8 Hz, Ar-H). 13C NMR (75 MHz, CDC13): 6 (ppm) 51.36, 66.88,
123.92, 126.06, 127.85, 127.91, 128.94, 129.34, 129.52, 130.71,
131.05, 132.50, 132.61, 133.95, 134.29, 145.23, 157.29, 181.51 (CO).
HRMS (ESI) calcd for C20H15N202SC1 [M] 382.8633; found [M+H]
383.0620.
Example 13
10-Chloro-6-thiomorpholino-12H-thiochromeno12,3-cpuinolin-
12-one (14)
Product 14 was prepared from 3 and thiomorpholine. The yellow
solid material was isolated in 86% yield (Rf= 0.77 at EA: n-hexane =
_
1: 4). Mp 219-220 C. 1H NMR (300 MHz, CDC13): 6 (ppm) 2.98 (4H,
t, J= 4.8 Hz, -NCH2-), 3.64 (4H, t, J= 5.1 Hz, -SCH2-), 7.62-7.84 (3H,
m, Ar-H), 7.73 (111, td, J= 8.4, 1.8 Hz, Ar-H), 8.06 (1H, dd, J= 8.1,
1.5 Hz, Ar-H), 8.57 (1H, dd, J= 1.8, 0.6 Hz, Ar-H), 9.61 (1H, dd, J-
7.8, 1.2 Hz, Ar-H). 13C NMR (75 MHz, CDC13): 6 (ppm) 27.06, 52.64,
114.95, 123.23, 125.40, 127.27, 127.31, 128.30, 128.68, 128.87,
130.55, 131.81, 131.96, 133.29, 133.70, 144.93, 157.43, 180.86 (CO).
HRMS (ESI) m/z calcd for C20H15N2S20C1+ [M]+ 398.0314, found
[M+1-1]+ 399.0420, [M+H+2]+ 401.0394.
Example 14
10-Chloro-6-(piperidin-l-y0-12H-thiochromeno12,3-clquinolin-
12-one (15)
Product 15 was prepared from 3 and piperidine. The yellow solid
material was isolated in 86% yield (Rf= 0.75 at EA). Mp 187-188 C.
28
CA 02866502 2014-10-03
. ,
. .
1H NMR (400 MHz, CDC13): 6 (ppm) 1.71-1.74 (2H, m, -CH2-), 1.88
(4H, p, J= 4.5 Hz, -CH2-), 3.31 (4H, t, J= 4.2 Hz, -NCH2-), 7.60-7.64
(2H, m, Ar-H), 7.61 (1H, d, J= 6.3 Hz, Ar-H), 7.69 (1H, td, J = 5.1,
1.2 Hz, Ar-H), 7.99 (1H, dd, J = 5.4, 0.6 Hz, Ar-H), 8.58 (1H, d, J =
5 1.5 Hz, Ar-H), 9.62 (1H, dd, J = 6.3, 0.6 Hz, Ar-H) 13C NMR (100
MHz, CDC13): 6 (ppm) 24.27, 25.89, 52.33, 123.47, 125.75, 127.28,
127.85, 128.56, 129.02, 129.23, 130.48, 131.62, 132.16, 132.33,
133.46, 134.67, 144.97, 158.50, 181.51 (CO). HRMS (ESI) calcd for
C21H17N20SC1 [M]+ 380.0750; found [M+H] + 381.0816.
10 Example 15
10-Chloro-6-(4-hydroxypiperidin-1-y1)-12H-thiochromeno[2,3-c
huinolin-12-one (16)
Product 16 was prepared from 3 and 4-hydroxypiperidine. The
gray-yellow solid material was isolated in 84% yield (Rf = 0.4 at EA:
15 n-hexane = 1: 1). Mp 224-225 C. 1H NMR (400 MHz, CDC13): 6
(ppm) 1.89 (1H, td, J = 7.2, 2.7 Hz, piperidine-CHa), 1.94 (1H, td, J =
6.9, 2.7 Hz, piperidine-CHa), 2.15-2.21 (2H, m, piperidine-CH,), 3.19
(2H, td, J = 8.4, 2.1 Hz, piperidine-NCHa), 3.60-3.65 (2H, m,
piperidine-NCHe), 4.01 (1H, sext, J = 3.0 Hz, piperidine-CH),
20 7.61-7.67 (3H, m, Ar-H), 7.71 (1H, td, J= 6.3, 1.2 Hz, Ar-H), 7.99
(1H, dd, J= 6.3, 0.6 Hz, Ar-H), 8.59 (1H, dd, J = 1.5, 0.6 Hz, Ar-H),
9.64 (1H, dd, J= 6.3, 0.6 Hz, Ar-H). 13C NMR (100 MHz, CDC13): 6
(ppm) 34.49, 48.90, 67.80, 123.62, 125.82, 127.54, 127.88, 128.64,
129.11, 129.36, 130.61, 131.29, 132.20, 132.47, 133.63, 134.43,
25 144.91, 157.83, 181.47 (CO). HRMS (ESI) calcd for C21H17N20SC1
[M]+ 396.0699; found [M+H]' 397.0757.
Example 16
29
CA 02866502 2014-10-03
6-(4-Benzylpiperidin-1-y1)-10-chloro-12H-thiochromeno[2,3-ch
uinolin-12-one (1 7)
Product 17 was prepared from 3 and 4-benzylpiperidine. The
yellow solid material was isolated in 90% yield (Rf = 0.57 at CH2C12:
n-hexane = 2: 1). Mp 184-185 C. 1H NMR (400 MHz, CDC13): 6
(ppm) 1.67 (2H, td, J= 9.3, 3.0 Hz, -CH2-), 1.79-1.89 (1H, m, -CH..),
1.88 (2H, d, J = 6.9 Hz, piperidine-CH2), 2.71 (2H, d, J= 5.1
piperidine-CH2), 3.00 (2H, td, J= 9.3, 1.2 Hz, -NCH2-), 3.65 (2H, d,J
= 9.3 Hz, -NCH2-), 7.20-7.25 (3H, m, Ar-H), 7.31-7.33 (2H, m, Ar-H),
7.60-7.72 (4H, m, Ar-H), 7.98 (1H, dd, J= 6.3, 0.6 Hz, Ar-H), 8.59
(1H, d, J= 1.8 Hz, Ar-H), 9.62 (1H, dd, J= 6.6, 0.6 Hz, Ar-H). 13C
NMR (100 MHz, CDC13): 6 (ppm) 32.21, 37.88, 43.30, 51.64, 123.52,
125.79, 125.99, 127.36, 127.87, 128.61, 129.09, 129.17, 129.29,
130.55, 131.49, 132.22, 132.41, 133.54, 134.62, 140.46, 144.98,
147.04, 158.25, 181.55 (CO). HRMS (ESI) calcd for C28H23N20SC1
[M]+ 471.0130; found [M+H] 471.1276.
Example 17
6-([1,4 '-Bipiperidini- 1 '-y1)- 10-chloro- 12H-thiochromeno [2,3 -eh
uinolin- 12-one (18)
Product 18 was prepared from 3 and 1,4'-bipiperidine. The yellow
solid material was isolated in 92% yield (Rf = 0.15 at EA: Me0H= 5:
1). Mp 187-189 C. 1H NMR (400 MHz, CDC13): 6 (ppm) 1.50-1.52
(2H, m, piperidine-H), 1.66-1.67 (3H, m, piperidine-H), 1.86-1.98 (2H,
qd, J = 12.4, 2.8 Hz, piperidine-H), 2.06 (2H, d, J = 11.6 Hz,
piperidine-H), 2.54 (1H, t, J= 10.8 Hz, piperidine-H), 2.65 (3H, br,
piperidine-H), 3.05 (2H, t, J= 12 Hz, piperidine-H), 3.73 (2H, d, J =
12.8 Hz, piperidine-H), 7.60-7.66 (3H, m, Ar-H), 7.70 (1H, td, J= 8.0,
CA 02866502 2014-10-03
. ,
. .
1.2 Hz, Ar-H), 7.98 (1H, d, J= 8.0 Hz, Ar-H), 8.58 (1H, s, Ar-H), 9.62
(1H, d, J = 8.8 Hz, Ar-H). 13C NMR (100 MHz, CDC13): 6 (ppm)
24.79, 26.36, 28.24, 50.45, 51.16, 62.40, 123.57, 125.80, 127.42,
127.86, 128.61, 129.08, 129.30, 130.52, 131.41, 132.18, 132.43,
5 133.56,
134.54, 144.92, 157.92, 181.47 (CO). HRMS (ESI) calcd for
C26H26N30SC1 [Mr- 463.1485; found [M+H]+ 464.1593.
Example 18
10-Chloro-6-(4-(3-(piperidin-4-y0propyl)piperidin-1-y1)-12H-thi
ochromeno[2,3-chuinolin-12-one (19)
10 Product 19 was
prepared from 3 and 1,3-di(piperidin-4-yl)propane.
The yellow solid material was isolated in 76% yield (Rf = 0.13 at
CH2C12). Mp 164-165 C. 11-1 NMR (400 MHz, CDC13): 6 (ppm) 1.16
(2H, qd, J= 11.6, 3.2 Hz, -CH2-), 1.20-1.28 (2H, m, -CH2-), 1.36-1.39
(4H, m, -CH2-), 1.51-1.58 (4H, m, -CH2-), 1.70 (2H, d, J= 13.6 Hz,
15 -CH2-), 1.87
(2H, d, J= 9.6 Hz, -CH2-), 2.43 (1H, br, -NH), 2.60 (2H,
td, J= 12.0, 2.0 Hz, -CH2-), 2.99 (2H, t, J= 11.2 Hz, -CH2-), 3.10 (2H,
d, J= 12 Hz, -CH2-), 3.64 (2H, d, J= 12.4 Hz, -CH2-), 7.59-7.65 (3H,
m, Ar-H), 7.68 (1H, td, J= 8.0, 1.2 Hz, Ar-H), 8.00 (1H, dd, J= 11.2,
1.2 Hz, Ar-H), 8.57 (1H, d, J= 1.6 Hz, Ar-H), 9.61 (1H, d, J= 8.4 Hz,
20 Ar-H). 13C NMR
(100 MHz, CDC13): 6 (ppm) 23.66, 32.40, 33.24,
35.73, 36.08, 36.86, 37.38, 46.58, 51.76, 123.49, 125.79, 127.30,
127.84, 128.57, 129.05, 129.26, 130.49, 131.55, 132.17, 132.36,
133.50, 134.64, 144.98, 158.34, 181.50 (CO). HRMS (EST) calcd for
C29H32N30SC1 [M]+ 505.1955; found [M+H] + 506.2004.
25 Example 19
10-Chloro-6-(pyrrolidin-1-y0-12H-thiochromeno[2,3-c]quinolin
-12-one (20)
31
CA 02866502 2014-10-03
. ,
. .
Product 20 was prepared from 3 and pyrrolidine. The solid
material was isolated in 86% yield (Rf= 0.56 at CH2C12: n-hexane = 1:
1). Mp 170-171 C. 1H NMR (300 MHz, CDC13): 6 (ppm) 2.05 (4H,
quin, J= 3.6 Hz, -CH2-), 3.76 (4H, t, J= 6.9 Hz, -NCH2-), 7.50(1H, td,
5 J= 7.2, 1.5
Hz, Ar-H), 7.61 (1H, d, J= 1.5 Hz, Ar-1-111), 7.65 (1H, td, J
=7.5, 1.5 Hz, Ar-H), 7.88 (1H, dd, J= 8.4, 1.5 Hz, Ar-H), 8.54 (1H, t,
J= 1.5 Hz, Ar-H), 9.44 (1H, dd, J= 8.7, 1.5 Hz, Ar-H). 13C NMR (75
MHz, CDC13): 6 (ppm) 24.89, 50.52, 121.90, 124.97, 125.17, 126.92,
127.33, 128.42, 128.84, 130.09, 131.77, 131.92, 133.10, 133.32,
144.73, 154.62, 159.91, 181.07 (CO). HRMS (ESI) calcd for
C20H15N20SC1 [M] 366.0594; found [M-FH] + 367.0659.
Example 20
10-chloro-6-(2-oxopiperidin-1-y1)-12H-thiochromeno[2,3-chuin
olin-12-one (TC-SCI-B-18) (21)
15 Product 21 was
a yellow solid material which was isolated in 89%
yield. Mp: 258-261 C.1H NMR (400 MHz, CDC13): 6 ppm. 1.25 (1H,
d, J = 4.8 Hz, piperidone-H), 2.44 (2H, quin, -CH2-), 2.61 (1H, s,
piperidone-H), 2.75 (2H, t, J = 8.4 Hz, -CH2-), 4.11-4.14 (2H, m,
-CH2-), 7.60-7.67 (2H, m, Ar-H), 7.78-7.81 (2H, m, Ar-H), 8.09-8.12
20 (1H, m, Ar-H),
8.60 (1H, d, J = 2.0 Hz, Ar-H), 9.72-9.75 (1H, m,
Ar-H). 13C NMR (100 MHz, CDC13): 6 ppm. 19.37, 31.47, 41.05,
49.14, 125.25, 126.08, 127.65, 129.29, 129.51, 129.80, 129.88, 130.74,
131.96, 132.26, 132.74, 133.47, 133.91, 145.05, 148.08, 176.14,
181.01.
25 Example 21
10-Chloro-6-methylamino-12H-thiochromeno[2,3-clquinolin-12
-one (N1)
32
CA 02866502 2014-10-03
Product N1 was prepared from 3 and methylamine. The pure
compound was obtained as a yellow solid (yield 92%) (Rf = 0.65 at
CH2C12). Mp 237-238 C (Me0H). 1H NMR (400 MHz, CDC13): 6
(ppm) 3.26 (3H, d, J= 4.8 Hz, -CH3), 4.92 (1H, d, J= 4.8 Hz, -NH-),
7.45 (1H, td, J= 11.2, 1.6 Hz, Ar-H), 7.58 (1H, d, J= 8.4 Hz, Ar-H),
7.69-7.65 (2H, m, Ar-H), 7.86 (1H, dd, J = 8.4, 0.8 Hz, Ar-H), 8.56
(1H,d, J = 1.6 Hz, Ar-H), 9.45 (1H, dd, J = 8.4, 1.2 Hz, Ar-H). 13C
NMR (100 MHz, CDC13): 6 (ppm) 29.38, 120.73, 123.68, 124.65,
125.87, 127.18, 127.49, 129.38, 129.51, 129.62, 131.04, 132.50,
132.53, 134.14, 145.64, 151.21, 180.96 (CO). HRMS (ESI) m/z calcd
for C17H11N20SC1 [M]+: 326.0281, found [M+Hr: 327.0356.
Example 22
10-Chloro-6-ethylamino-12H-thiochromeno[2,3-dquinolin-12-
one (N2)
Product N2 was prepared from 3 and ethylamine. The pure
compound was obtained as a yellow solid (yield 91%) (Rf = 0.75 at
CH2C12). Mp 204-205 C (Me0H). 1H NMR (400 MHz, CDC13): 6
(ppm) 1.41 (3H, t, J= 7.2 Hz, -CH3), 3.75 (2H, q, J= 1.6 Hz, -C1-12),
4.81 (1H, br, -NH-), 7.44 (1H, td, J = 8.4, 1.6 Hz, Ar-H), 7.58-7.64
(3H, m, Ar-H), 7.83 (1H, d, J= 8.4 Hz, Ar-H), 8.56 (1H, d, J= 1.6 Hz,
Ar-H), 9.44 (1H, d, J= 8.4 Hz, Ar-H). 13C NMR (100 MHz, CDC13): 6
(ppm) 14.79, 37.28, 120.70, 123.50, 124.58, 125.84, 127.18, 127.47,
129.39, 129.47, 129.66, 131.08, 132.50, 132.53, 134.12, 145.67,
150.55, 181.00 (CO). HRMS (ESI) In/z calcd for C18H13N20SC1 [M]:
340.0437, found [M+H]': 341.0493.
Example 23
10-Chloro-6-propylamino-12H-thiochromeno[2,3-dquinolin-12
33
CA 02866502 2014-10-03
, .
. .
-one (N3)
Product N3 was prepared from 3 and propylamine. The pure
compound was obtained as a yellow solid (yield 85%) (Rf = 0.82 at
CH2C12). Mp 178-179 C (Me0H). 1H NMR (400 MHz, CDC13): 6
5 (ppm) 1.09
(3H, t, J = 7.2 Hz, -CH3), 1.81 (2H, sext, J = 7.2 Hz,
-CH2-), 3.69 (2H, q, J = 7.2 Hz, -NCH2-), 4.87 (1H, br, -NH-), 7.44
(1H, td, J= 8.0, 1.2 Hz, Ar-H), 7.58-7.64 (3H,m, Ar-H), 7.82 (1H, d, J
= 8.0 Hz, Ar-H), 8.56 (1H,d, J= 1.2 Hz, Ar-H), 9.44 (1H, d, J= 8.8
Hz, Ar-H). 13C NMR (100 MHz, CDC13): 6 (ppm) 11.68, 22.64, 44.13,
120.67, 123.53, 124.53, 125.82, 127.15, 127.46, 129.38, 129.46,
129.65, 131.05, 132.49, 134.10, 145.66, 150.62, 181.02 (CO). HRMS
(ESI) m/z calcd for C19ll15N20SC1 [Mr: 354.0594, found [M+H]':
355.0651.
Example 24
15 6-(Butylamino)-
10-chloro-12H-thiochromeno[2,3-chuinolin-12
-one (N4)
Product N4 was prepared from 3 and butylamine. The pure
compound was obtained as a yellow solid (yield 91%) (Rf = 0.85 at
CH2C12). Mp 147-149 C (Me0H). 1H NMR (400 MHz, CDC13): 8
20 (ppm) 1.03
(3H, t, J = 7.2 Hz,-CH3), 1.53 (2H, sext, J = 7.2 Hz,
-CH2-), 1.76 (2H, quin, J= 7.2 Hz, -CH2-), 3.71 (2H, q, J= 6.8 Hz,
-NCH2-), 4.83 (1H, br, -NH-), 7.43 (1H, td, J = 7.6, 1.2 Hz, Ar-H),
7.57-7.64 (3H, m, Ar-H), 7.82 (1H, d, J= 8.4 Hz, Ar-H), 8.55 (1H, d,
J = 1.6 Hz, Ar-H), 9.43 (111, d, J = 8.4 Hz, Ar-H). 13C NMR (100
25 MHz, CDC13): 8
(ppm) 13.97, 20.36, 31.54, 42.12, 120.67, 123.53,
124.52, 125.84, 127.16, 127.46, 129.38, 129.46, 129.65, 131.05,
132.48, 132.52, 134.11, 145.68, 150.62, 181.00 (CO). HRMS (ESI)
34
CA 02866502 2014-10-03
. .
m/z calcd for C201-117N20SC1 [Mr: 368.0750, found {M+H}:
369.0846.
Example 25
10-Chloro-6-isobutylamino-12H-thiochromeno[2,3-c]quinolin-
12-one (N5)
Product N5 was prepared from 3 and isobutylamine. The pure
compound was obtained as a yellow crystal (yield 61%) (Rf = 0.85 at
CH2C12). Mp 159-160 C (Me0H). 11-1 NMR (400 MHz, CDC13): 6
(ppm) 1.08 (6H, d, J= 6.8 Hz,-CH3), 2.10 (1H, sep, J= 6.8 Hz,-CH..),
3.56 (211, t, J= 6.4 Hz, -CH2_), 4.94 (111, br, -NH), 7.44(1H, t, J= 7.2
Hz, Ar-H), 7.59-7.64 (3H, m, Ar-H), 7.82 (1H, d, J= 8.4 Hz, Ar-H),
8.57 (1H, dd, J= 2.0, 0.8 Hz, Ar-H), 9.43 (1H, d, J= 8.4 Hz, Ar-H).
13C NMR (100 MHz, CDC13): 6 (ppm) 20.50, 28.16, 49.72, 120.67,
123.54, 124.51, 125.82, 127.13, 127.47, 129.39, 129.47, 129.69,
131.02, 132.50, 134.11, 138.34, 145.62, 150.68, 181.02 (CO). HRMS
(ESI) m/z calcd for C20H17N20SC1 [Mr: 368.0750, found [M+Hr:
369.0825.
Example 26
10-Chloro-6-(pentan-3-ylamino)-12H-thiochromenoI2,3-elquin
olin-12-one (N6)
Product N6 was prepared from 3 and 3-aminopentane. The pure
compound was obtained as a light yellow crystal (yield 65%) (Rf =
0.87 at CH2C12). Mp 160-161 C (Me0H). 111 NMR (400 MHz,
CDC13): 6 (ppm) 0.92 (6H, t, J= 7.6 Hz, -CH3), 1.69 (4H, quin, J=
6.0 Hz, -CH2-), 4.34 (1H, sext, J= 7.2 Hz, -CH-), 6.58 (111, d, J= 8.0
Hz, Ar-H), 7.36(1H, t, J = 8.0 Hz, Ar-H), 7.58 (1H, t, J = 8.0 Hz,
Ar-H), 7.65 (1H, d, J= 8.0 Hz, Ar-H), 7.90 (1H, dd, J= 8.4, 2.4 Hz,
CA 02866502 2014-10-03
õ
Ar-H), 8.01 (1H, d, J= 8.4 Hz, Ar-H), 8.40 (1H, d, J = 2.4 Hz, Ar-H),
9.34 (1H, d, J= 8.8 Hz, Ar-H). 13C NMR (100 MHz, CDC13): 6 (ppm)
11.21, 26.86, 54.41, 120.16, 123.92, 125.13, 125.81, 127.05, 128.49,
129.02, 129.37, 129.73, 132.17, 132.55, 133.22, 133.29, 145.69,
5 151.64, 180.85
(CO). HRMS (ESI) m/z calcd for C2Ifl19N20SC1 [M]:
382.0907, found [M+H]: 383.0994, [M-Hp381.0851.
Example 27
10-Chloro-642-(dimethylamino)ethyl)amino)-12H-thiochrome
no[2,3-dquinolin-12-one (N7)
10 Product N7 was prepared from 3 and
/V,N-dimethylethylenediamine. The pure compound was obtained as a
yellow crystal (yield 76%) (Rf= 0.82 at EA: MeOH: ammonia water=
10:5:1). Mp 156-157 C (Me0H). 1H NMR (400 MHz, CDC13): 6
(13Prn) 2.36 (6H, s, -N(CH3)2), 2.69 (2H, t, J = 6.0 Hz, -CH2N-), 3.57
15 (2H, q, J= 5.6
Hz, -NCH2-), 5.86 (1H, br, -NH), 7.44 (1H, td, J = 8.0,
1.6 Hz, Ar-H), 7.62 (2H, td, J = 7.6, 1.6 Hz, Ar-H), 7.65 (1H, dd, J =
8.0, 0.8 Hz, Ar-H), 7.82 (1H, dd, J = 8.4, 1.2 Hz, Ar-H), 8.58 (1H, dd,
J = 1.6, 0.4 Hz, Ar-H), 9.46 (1H, dd, J= 8.8, 1.2 Hz, Ar-H). 13C NMR
(100 MHz, CDC13): 6 (ppm) 39.53, 45.28, 57.57, 120.71, 123.46,
20 125.90, 127.02, 127.27, 127.57, 129.35, 129.43, 129.56, 131.46,
132.43, 132.56, 134.02, 145.72, 150.90, 181.05. (CO). HRMS (ESI)
m/z calcd for C20H18N30SC1 [M]: 383.0859, found [M+H]:
384.0925.
Example 28
25 10-Chloro-6((2-
(diethylamino)ethyl)amino)-12H-thiochromen
42,3-cpuinolin-12-one (N8)
Product N8 was prepared from 3 and N,N-diethylethylenediamine.
36
CA 02866502 2014-10-03
The pure compound was obtained as a yellow crystal (yield 86%) (Rf
= 0.8 at EA: MeOH: ammonia water= 10:5:1). Mp 152-153 C
(Me0H). 1H NMR (400 MHz, CDC13): 6 (ppm) 1.13 (6H, t, J = 7.2
Hz,-CH3), 2.64 (4H, q, J = 6.8 Hz,-NCH2-), 2.82 (2H, t, J = 6.0
Hz,-CH2N-), 3.70 (2H, q, J = 5.2 Hz, -NCH2-), 6.08 (1H, br, -NH-),
7.43(1H, t, J = 7.2 Hz, Ar-H), 7.59-7.64 (311, m, Ar-H), 7.81 (1H, d, J
= 8.4 Hz, Ar-H), 8.57 (1H, d, J= 1.2 Hz, Ar-H), 9.45 (1H, d, J = 8.4
Hz, Ar-H). 13C NMR (100 MHz, CDC13): 6 (ppm) 12.34, 39.52, 46.81,
50.89, 120.64, 124.25, 124.34, 125.89, 126.96, 127.63, 129.32, 129.41,
129.53, 131.49, 132.37, 132.55, 133.98, 145.79, 150.96, 181.06 (CO).
HRMS (ESI) m/z calcd for C22H22N30SC1 [M]: 411.1172, found
[M+H]: 412.1262.
Example 29
10-Chloro-6-(2-ethanolamino)-12H-thiochromeno[2,3-c]quinol
in-12-one (N9)
Product N9 was prepared from 3 and ethanolamine. The pure
compound was obtained as a yellow crystal (yield 77%) (Rf = 0.65 at
EA). Mp 190-192 C (Me0H). 1H NMR (400 MHz, CDC13): 6 (ppm)
3.93 (2H, q, J = 4.4 Hz, -NCH2-), 4.00 (2H, t, J = 4.4 Hz, -CH20-),
4.23 (1H, br, -OH), 5.45 (1H, br, -NH), 7.48 (1H, td, J = 8.0, 1.6 Hz,
Ar-H), 7.62-7.68 (3H, m, Ar-H),7.81 (1H, dd, J =7.6, 0.8 Hz, Ar-H),
8.58 (1H, dd, J = 1.6, 0.4 Hz, Ar-H), 9.45 (1H, dd, J = 8.4, 0.8 Hz,
Ar-H). 13C NMR (100 MHz, CDC13): 6 (ppm) 45.88, 63.59, 120.97,
123.67, 125.12, 125.92, 126.60, 127.50, 129.48, 129.83, 130.15,
130.91, 132.55, 132.69, 134.35, 144.65, 151.32, 180.87 (CO). HRMS
(ESI) m/z calcd for C18H13N202SC1 [M]: 356.8260, found [M+H]:
357.0476, [M+H+2]: 359.0455.
37
CA 02866502 2014-10-03
. .
= 1
Example 30
10-Chloro-6-(3-propanolamino)-12H-thiochromeno[2,3-clquin
olin-12-one (N10)
Product N10 was prepared from 3 and 3-amino-1-propanol. The
5 pure compound
was obtained as a yellow solid (yield 94%) (Rf= 0.66
at EA). Mp 201-202 C (Me0H). 1H NMR (400 MHz, CDC13): 6
(PPin) 1.94 (2H, p, J = 6.0 Hz, -CH2-), 3.72 (2H, t, J = 5.2 Hz,
-NCH2-), 3.93 (2H, q, J= 6.0 Hz, -CH20-), 4.41 (1H, br, -OH), 5.38
(1H, t, J = 5.2 Hz, -NH-), 7.45 (1H, td, J = 7.6, 1.2 Hz, Ar-H),
10 7.58-7.65 (3H,
m, Ar-H), 7.78 (1H, dd, J= 8.4, 0.8 Hz, Ar-H), 8.56
(1H, dd, J= 2.0, 0.4 Hz, Ar-H), 9.42 (1H, dd, J= 8.4, 1.2 Hz, Ar-H).
13C NMR (100 MHz, CDC13): 6 (ppm) 33.23, 38.94, 59.25, 120.72,
123.44, 124.84, 125.94, 126.32, 127.45, 129.44, 129.92, 130.11,
130.84, 132.49, 132.64, 134.31, 144.83, 151.33, 180.88 (CO). HRMS
15 (ESI) m/z
calcd for C19H15N202SC1 [M]+: 370.0543, found [M+H]:
371.0622.
Example 31
10-Chloro-6-(5-pentanolamino)-12H-thiochromeno[2,3-c]quin
olin-12-one (N11)
20 Product N11
was prepared from 3 and 5-amino-1-pentanol.The
pure compound was obtained as a yellow solid (yield 91%) (Rf = 0.7
at EA). Mp 158-160 C (Me0H). 111 NMR (400 MHz, CDC13): 6
(ppm) 1.40 (1H, br, -0H),1.49-1.62 (2H, m, -CH2-), 1.71 (2H, quin,
-CH2-), 1.83 (2H, quin, -CH2-), 3.74 (4H, quin, -CH2-), 4.91 (1H, br,
25 -NH), 7.45(1H,
td, J= 7.6, 1.2 Hz, Ar-H), 7. 59 (3H, m, Ar-H), 7.83
(1H, d, J= 8.4 Hz, Ar-H), 8.57 (1H, d, J= 1.2 Hz, Ar-H), 9.44 (1H, d,
J= 8.4 Hz, Ar-H). 13C NMR (100 MHz, CDC13): 6 (ppm) 23.39, 29.17,
38
CA 02866502 2014-10-03
= i
32.40, 42.24, 62.83, 120.69, 123.53, 124.60, 125.84, 127.11, 127.48,
129.40, 129.51, 129.71, 131.02, 131.79, 132.53, 134.14, 145.60,
150.58, 181.02 (CO). HRMS (ESI) m/z calcd for C211119N202SC1 [M]:
398.0856, found [M+H]: 399.0914.
Example 32
10-Chloro-641-hydroxybutan-2-y0amino)-12H-thiochromenof
2,3-dquinolin-12-one (N12)
Product N12 was prepared from 3 and 2-amino-1-butanol.The
pure compound was obtained as a yellow solid (yield 94%) (Rf = 0.8
at EA). Mp 203-204 C (Me0H). 11-1 NMR (400 MHz, CDC13): 6
(PPm) 1.21 (3H, t, J = 7.6 Hz, -CH3), 1.71-1.88 (2H, m, -CH2-), 3.79
(1H, dd, J =11.2, 1.6 Hz, -CH2-), 3.99 (1H, dd, J = 11.2, 2.8 Hz,
-CH2-), 4.34 (1H, quin, J= 11.2 Hz, -NCH-), 4.59 (1H, br, -OH), 5.02
(1H, d, J = 6.0 Hz, -NH-), 7.44(1H, td, J = 8.0, 1.2 Hz, Ar-H), 7.56
(1H, d, J = 8.4 Hz, Ar-H), 7.59 (1H, td, J = 7.6, 1.2 Hz, Ar-H), 7.61
(1H, d, J= 8.4 Hz, Ar-H), 7.74 (1H, d, J= 8.4 Hz, Ar-H), 8.52 (1H, d,
J= 2.0 Hz, Ar-H), 9.40 (1H, dd, J= 7.6, 1.2 Hz, Ar-H). 13C NMR (100
MHz, CDC13): 6 (ppm) 10.95, 24.95, 56.89, 67.08, 120.83, 123.68,
124.98, 125.88, 126.51, 127.40, 129.39, 129.75, 130.02, 130.81,
132.38, 132.62, 134.28, 144.53, 151.09, 180.75 (CO). HRMS (ESI)
m/z calcd for C20H17N202SC1 [M]: 384.0699, found [M+11]:
385.0790.
Example 33
10-Chloro-644-methylpentan-2-y0amino)-12H-thiochromenof
2,3-dquinolin-12-one (N13)
Product N13 was prepared from 3 and
4-methylpentan-2-amine.The pure compound was obtained as a
39
CA 02866502 2014-10-03
,
yellow solid (yield 94%) (Rf = 0.9 at CH2C12). Mp 176-177 C
(Me0H).
NMR (400 MHz, CDC13): 6 (ppm) 0.98 (3H, d, J= 6.8
Hz, -CH3), 1.03 (3H, d, J= 6.8 Hz, -CH3), 1.35 (3H, d, J= 6.4 Hz,
-CH3), 1.45 (1H, quin, J= 6.4 Hz, -CH2-), 1.66 (1H, quin, J= 6.8 Hz,
-CH2-), 1.80 (1H, sep, J = 6.8 Hz, -CH-), 4.63 (1H, br, -NH),
4.63-4.66 (1H, m, -CH-), 7.43 (1H, td, J = 7.6, 1.2 Hz, Ar-H),
7.58-7.63 (3H, m, Ar-H), 7.81 (1H, d, J= 8.4 Hz, Ar-H), 8.56 (1H, d,
J = 1.2 Hz, Ar-H), 9.43 (1H, d, J= 8.4 Hz, Ar-H). 13C NMR (100
MHz, CDC13): 6 (ppm) 21.38, 22.87, 22.92, 25.40, 45.73, 46.80,
120.55, 123.42, 124.36, 125.80, 127.21, 127.44, 129.38, 129.71,
131.08, 132.47, 132.52, 134.07, 145.76, 150.01, 181.05 (CO). HRMS
(ESI) m/z calcd for C22H19N20SC1 [M]: 396.1063, found [M+H]:
397.1142.
Example 34
6((2-Aminoethyl)amino)-1O-chloro-12H-thiochromeno[2,3-ch
uinolin-12-one (N14)
Product N14 was prepared from 3 and 1,2-diaminoethane. The
pure compound was obtained as a yellow solid (yield 90%) (Rf = 0.6
at EA: MeOH: ammonia water = 10: 5: 1). Mp 193-194 C (Me0H).
11-1 NMR (400 MHz, DMSO-d6): 6 (ppm) 2.90 (2H, t, J = 6.0 Hz,
-CH2-), 3.59 (2H, t, J= 6.0 Hz, -CH2-), 7.36 (1H, t, J= 8.0 Hz, Ar-H),
7.59(1H, t, J= 8.0 Hz, Ar-H), 7.66 (111, d, J=8.0 Hz, Ar-H), 7.85 (1H,
d, J= 7.2 Hz, Ar-H), 7.96 (1H, d, J¨ 8.8 Hz, Ar-H), 8.35 (1H, br,
Ar-H), 9.32 (1H, d, J = 8.4 Hz, Ar-H). 13C NMR (100 MHz,
DMSO-d6): 6 (ppm) 40.79, 45.06, 120.24, 124.08, 125.31, 125.84,
127.00, 128.39, 128.83, 129.32, 129.69, 132.06, 132.33, 133.12,
133.26, 145.58, 151.52, 180.65 (CO). HRMS (ESI) m/z calcd for
CA 02866502 2014-10-03
= .
C18H14N30SC1 [MI*: 355.0546, found [M-FH]+: 356.0641.
Example 35
10-Chloro-642-((2-hydroxyethyl)amino)ethyl)amino)-12H-thio
chromeno[2,3-dquinolin-12-one (N15)
Product N15 was prepared from 3 and
N-(2-hydroxyethyl)ethylenediamine. The pure compound was
obtained as a yellow solid (yield 58%) (Rf = 0.63 at EA: MeOH:
ammonia water = 10: 5: 1). Mp 141-143 C (Me0H). 111 NMR (400
MHz, DMSO-d6): 8 (ppm) 2.69 (2H, t, J= 5.6 Hz, -CH2-), 2.90 (2H, t,
J= 6.0 Hz, -CH2-), 3.51 (2H, t, J= 5.6 Hz, -CH2-), 3.65 (2H, t, J= 6.0
Hz, -CH2-), 7.10 (1H, br, -NH-), 7.32 (1H, t, J= 7.2 Hz, Ar-H), 7.55
(1H, t, J= 7.2 Hz, Ar-H), 7.62 (1H, d, J= 8.4 Hz, Ar-H), 7.76 (1H, t, J
= 7.2 Hz, Ar-H), 7.86 (1H, d, J= 8.4 Hz, Ar-H), 8.25 (1H, d, J = 2.0
Hz, Ar-H), 9.26 (1H, d, J = 8.4 Hz, Ar-H). 13C NMR (100 MHz,
DMSO-d6): 6 (ppm) 41.90, 48.16, 51.80, 60.72, 120.13, 123.96,
125.14, 125.81, 126.94, 128.26, 128.65, 129.16, 129.60, 131.82,
132.11, 132.96, 133.19, 145.50, 151.35, 180.45 (CO). HRMS (ESI)
m/z calcd for C20H18N302SC1 [Mr: 399.8938, found [M+Hr:
400.0880.
Example 36
1 0-Chloro-642-morph olinoethyl)amino)- 1 2H-thiochromeno [2,
3-chuinolin-12-one (N16)
Product N16 was prepared from 3 and
4-(2-aminoethyl)morpholine. The pure compound was obtained as a
yellow solid (yield 87%) (Rf = 0.48 at EA). Mp 189-190 C (Me0H).
11-1 NMR (400 MHz, CDC13): 6 (ppm) 2.63 (4H, br, -CH2-), 2.81 (2H,
br, -CH2-), 3.81 (6H, br, -CH2-), 5.92 (111, br, -NH-), 6.70 (2H, d, J=
41
CA 02866502 2014-10-03
8.4 Hz, Ar'-H), 7.45 (1H, td, J= 7.8, 1.6 Hz, Ar-H), 7.60-7.64 (3H, m,
.Ar-H), 7.81 (1H, d, J= 8.4 Hz, Ar-H), 8.57 (1H, s, Ar-H), 9.46 (1H,
dd, J = 8.8, 1.2 Hz, Ar-H). 13C NMR (100 MHz, CDC13): 8 (ppm)
38.28, 53.31, 56.52, 67.13, 120.73, 123.94, 124.58, 125.91, 127.01,
127.56, 129.37, 129.49, 129.64, 131.27, 132.47, 132.54, 134.10,
145.66, 150.76, 180.99 (CO). HRMS (ESI) m/z calcd for
C22H20N302SC1 [M]+: 425.0965, found [M+H]: 426.1058, [M-HI:
424.0885.
Example 37
10-Chloro-643-(dimethylamino)propyl)amino)-12H-thiochrom
eno[2,3-chuinolin-12-one (N17)
Product N17 was prepared from 3 and
3-(dimethylamino)-1-propylamine. The pure compound was obtained
as a yellow crystal (yield 43%) (Rf = 0.71 at EA: MeOH: ammonia
water = 10: 5: 1). Mp 194-195 C (Me0H). 1H NMR (400 MHz,
CDC13): 8 (ppm) 1.92 (2H, quin, J =6.0 Hz, -CH2-), 2.41 (6H, s,
-N(CH3)2), 2.60 (2H, t, J= 5.6 Hz, -CH2N-), 3.81 (2H, q, J= 5.6 Hz,
-NCH2-), 7.95 (1H, br, -NH), 7.40 (1H, td, J = 7.6, 1.6 Hz, Ar-H),
7.56-7.63 (4H, m, Ar-H), 7.80 (1H, d, J= 8.4 Hz, Ar-H), 8.57 (1H, d,
J= 2.4 Hz, Ar-H), 9.44 (1H, dd, J= 7.6, 0.8 Hz, Ar-H). 13C NMR (100
MHz, CDC13): 8 (ppm) 24.83, 43.64, 45.68, 59.72, 120.46, 123.97,
124.56, 125.84, 126.83, 127.54, 129.32, 129.48, 131.84, 132.29,
132.56, 133.86, 146.01, 151.27, 181.14 (CO). HRMS (ESI) m/z calcd
for C2II-120N30SC1 [Mr: 397.1016, found [M+H]: 398.1072.
Example 38
10-Chloro-643-(diethylamino)propyl)amino)-12H-thiochrome
no[2,3-chuinolin-12-one (N18)
42
CA 02866502 2014-10-03
Product N18 was prepared from 3 and
3-(diethylamino)-1-propylamine. The pure compound was obtained as
a yellow acicular crystal (yield 70%) (Rf = 0.68 at EA: MeOH:
ammonia water = 10: 5: 1). Mp 142-143 C (MeOH). 1H NMR (400
MHz, CDC13): 6 (ppm) 1.15 (6H, t, J= 6.8 Hz, -CH3), 1.91 (2H, quin,
J = 6.0 Hz, -CH2-), 2.66-2.72 (6H, m, -NCH2-), 3.81 (2H, q, J = 4.8
Hz, -NCH2-), 7.40 (1H, td, J= 7.2, 1.2 Hz, Ar-H), 7.55-7.58 (1H, dd,J
= 8.4, 3.6 Hz, Ar-H), 7.60-7.64 (2H, m, Ar-H), 7.81 (1H, d, J= 8.0 Hz,
Ar-H), 7.93 (1H, br, Ar-H), 8.58 (1H,t,J= 2.0 Hz, Ar-H), 9.45 (1H, dd,
J= 8.4, 0.8 Hz, Ar-H). 13C NMR (100 MHz, CDC13): 6 (ppm) 11.44,
24.71, 44.25, 47.09, 53.57, 120.45, 123.99, 124.50, 125.86, 126.87,
127.46, 129.34, 129.43, 131.85, 132.32, 132.57, 133.88, 146.04,
151.30, 181.15 (CO). HRMS (ESI) m/z calcd for C23H24N3S0C1 [M]+:
425.1329, found [M+Hr: 426.1396, [M-Hf: 424.1284.
Example 39
10-Chloro-64342-hydroxyethyl)amino)propyl)amino)-12H-th
iochromeno[2,3-clquinolin-12-one (N19)
Product N19 was prepared from 3 and
N-(2-Hydroxyethyl)-1,3-diaminopropane. The pure compound was
obtained as a brown solid (yield 75%) (Rf = 0.65 at EA: MeOH:
ammonia water = 10: 5: 1). Mp 65-67 C (MeOH). 1H NMR (400
MHz, CDC13): 6 (ppm) 1.89 (2H, quin, J= 6.0 Hz, -CH2-), 2.15 (2H,
br, -OH & -NH-), 2.85 (4H, quin, -CH2-), 3.74 (2H, t, J = 6.0 Hz,
-CH2-), 3.80 (2H, t, J= 5.2 Hz, -CH2-), 6.53 (1H, br, -NH-), 7.39 (1H,
td, J= 7.6, 0.8 Hz, Ar-H), 7.44 (1H, d, J= 8.8 Hz, Ar-H), 7.50 (111, dd,
J= 8.4, 2.4 Hz, Ar-H), 7.58 (1H, td, J= 7.2, 1.2 Hz, Ar-H), 7.76 (1H,
d, J= 8.0 Hz, Ar-H), 8.46 (1H, d, J= 2.0 Hz, Ar-H), 9.39 (1H, d, J=
43
CA 02866502 2014-10-03
. .
= .
8.4 Hz, Ar-H). 13C NMR (100 MHz, CDC13): 6 (ppm) 28.37, 42.23,
48.74, 51.65, 61.42, 120.48, 123.99, 124.22, 125.85, 126.88, 127.42,
129.13, 129.33, 129.38, 131.23, 132.21, 132.33, 133.95, 145.69,
150.82, 180.92 (CO). HRMS (ESI) m/z calcd for C211-120N302SC1 [M]:
5 413.0965, found [M+H]: 414.1053, [M+H+2]: 416.1037.
Example 40
10-Chloro-642,3-dihydro-1H-inden-2-y0amino)-12H-thiochro
meno[2,3-elquinolin-12-one (N20)
Product N20 was prepared from 3 and 2-aminoindane. The pure
10 compound was obtained as a brown solid (yield 65%) (Rf = 0.7 at
CH2C12: n-hexane= 2: 1). Mp 251-252 C (Me0H). 1H NMR (400
MHz, CDC13): 6 (ppm) 3.02 (1H, d, J= 5.2 Hz, indane-H), 3.06 (1H, d,
J= 5.2 Hz, indane-H), 3.59 (1H, d, J= 7.2 Hz, indane-H), 3.63 (1H, d,
J= 7.2 Hz, indane-H), 5.10 (1H, d, J= 6.8 Hz, -NH), 5.23 (1H, q, J-
15 5.2 Hz, indane-H), 7.21-7.25 (211, m, Ar'-H), 7.28-7.31 (2H, m,
Ar'-H), 7.47 (1H, td, J= 6.8, 1.2 Hz, Ar-H), 7.58 (1H, d, J= 8.4 Hz,
Ar-H), 7.61-7.67 (1H, td, J= 6.8, 1.2 Hz, Ar-H), 7.87 (1H, d, J= 7.6
Hz, Ar-H), 8.57 (1H, d, J= 2.0 Hz, Ar-H), 9.46 (1H, dd, J= 8.8, 0.8
Hz, Ar-H). 13C NMR (100 MHz, CDC13): 6 (ppm) 40.41, 53.27,
20 120.79, 123.60, 124.81, 124.93, 125.84, 126.76, 127.37, 127.46,
129.38, 129.51, 129.71, 131.02, 132.50, 132.54, 134.12, 141.29,
145.56, 150.20, 181.00 (CO). HRMS (ESI) m/z calcd for
C25H17N20SC1 [M]: 428.0750; found [M+H]: 429.0822.
Example 41
25 10-Chloro-6-(cyclohexylamino)-12H-thiochromeno[2,3-ciquino
lin-12-one (N21)
Product N21 was prepared from 3 and cyclohexylamine. The
44
CA 02866502 2014-10-03
g
pure compound was obtained as a brown solid (yield 91%) (Rf= 0.7 at
CH2C12: n-hexane= 2: 1). Mp 196-197 C (Me0H). 1H NMR (400
MHz, CDC13): 6 (ppm) 1.25-1.40 (4H, m, cyclohexylamine-CH2),
1.49-1.60 (2H, m, cyclohexylamine-CH2), 1.70-1.74 (2H, m,
cyclohexylamine-CH2), 1.79-1.84 (2H, m, cyclohexylamine-CH2),
2.21 (2H, dd, J= 8.8, 3.2 Hz, cyclohexylamine-CH2), 4.30 (1H, sep, J
= 3.6 Hz, cyclohexylamine-CH), 4.72 (1H, d, J = 6.8 Hz, -NH-),
7.41(1H, t, J= 8.0 Hz, Ar-H), 7.51-62 (3H, m, Ar-H), 7.79 (1H, d, J=
8.0 Hz, Ar-H), 8.51 (1H, d, J= 1.6 Hz, Ar-H), 9.41 (1H, d, J= 8.4 Hz,
Ar-H). 13C NMR (100 MHz, CDC13): 6 (ppm) 24.94, 25.92, 33.10,
50.26, 120.51, 123.50, 124.34, 125.77, 127.11, 127.37, 129.27, 129.35,
129.60, 131.01, 132.37, 132.41, 134.00, 145.66, 149.75, 180.95 (CO).
HRMS (ESI) m/z calcd for C22Hi9N2OSC1 [M]+: 394.0907; found
[M+H]: 395.0991.
Example 42
641-Benzylpiperidin-4-y0amino)-10-chloro-12H-thiochromen
o[2,3-c]quinolin-12-one (N22)
Product N22 was prepared from 3 and 1-benzylpiperidin-4-amine.
The pure compound was obtained as a brown solid (yield 62%) (Rf --
0.77 at EA). Mp 194-196 C (Me0H). 1H NMR (400 MHz, CDC13): 8
(ppm) 1.62-1.72 (2H, m, piperidine-H), 2.24 (2H, d, J = 13.2 Hz,
piperidine-H), 2.32 (2H, t, J= 11.2 Hz, piperidine-H), 2.92 (211, d, J=
11.6 Hz, piperidine-H), 3.59 (2H, s, -CH2-), 4.35 (1H, sext, J= 6.4 Hz,
piperidine-CH), 4.75 (1H, d, J = 7.2 Hz, -NH), 7.26-7.30 (1H, m,
Ar'-H), 7.36-7.38 (4H, m, Ar'-H), 7.44 (1H, td, J= 7.6, 0.8 Hz, Ar-H),
7.59-7.64 (3H, m, Ar-H), 7.80 (1H, d, J= 7.6 Hz, Ar-H), 8.56 (1H, d,
J= 1.6 Hz, Ar-H), 9.43 (1H, d, J=8.8 Hz, Ar-H). 13C NMR (100 MHz,
CA 02866502 2014-10-03
=
CDC13): .3 (ppm) 32.26, 48.63, 52.35, 63.22, 120.63, 123.47, 124.57,
125.81, 127.07, 127.12, 127.47, 128.25, 129.21, 129.35, 129.44,
129.74, 130.99, 132.46, 132.52, 134.10, 138.37, 145.57, 149.76,
181.00 (CO). HRMS (ESI) m/z calcd for C28H24N30SC1 [M]+:
485.1329; found [M+H]: 486.1379.
Example 43
10-Chloro-6-((thiophen-2-ylmethyl)amino)-12H-thiochromeno[
2,3-dquinolin-12-one (N23)
Product N23 was prepared from 3 and 2-thiophenemethylamine.
The pure compound was obtained as a brown solid (yield 78%) (Rf =
0.7 at CH2C12: n-hexane = 2: 1). Mp 178-180 C (Me0H). 1H NMR
(400 MHz, CDC13): 6 (ppm) 5.07 (111, d, J = 5.2 Hz, -NCH2-), 5.17
(1H, br, -NH-),7.00 (1H, t, J= 4.4 Hz, thiophene-H), 7.16 (1H, d, J =
3.2 Hz, thiophene-H), 7.25 (1H, d, J= 0.8 Hz, thiophene-H), 7.47(1H,
t, J= 8.0 Hz, Ar-H), 7.52 (1H, d, J= 8.4 Hz, Ar-H), 7.58 (1H, d, J =
8.4 Hz, Ar-H), 7.65 (1H, t, J= 7.6 Hz, Ar-H), 7.89 (1H, d, J = 8.0 Hz,
Ar-H), 8.53 (1H, s, Ar-H), 9.46 (1H, d, J = 8.4 Hz, Ar-H). 13C NMR
(100 MHz, CDC13): 60 (ppm) 41.14, 121.06, 123.58, 125.05, 125.47,
125.89, 126.47, 126.73, 127.29, 127.45, 129.33, 129.57, 129.71,
130.94, 132.39, 132.51, 134.12, 141.36, 145.20, 149.82, 180.80 (CO).
HRMS (ESI) m/z calcd for C21H13N20S2C1 [M]: 408.0158; found
[M+H]: 409.0251, [M-Hf: 407.0085.
Example 44
10-Chloro-6-((cyclohexylmethyl)amino)-12H-thiochromenoI2,3
-dquinolin-12-one (N24)
Product N24 was prepared from 3 and cyclohexylmethanamine.
The pure compound was obtained as a brown solid (yield 79%) (Rf =
46
CA 02866502 2014-10-03
=
0.7 at CH2C12: n-hexane = 2: 1). Mp 165-166 C (Me0H). 1H NMR
(400 MHz, CDC13): 6 (ppm) 1.07 (1H, d, J = 11.2 Hz,
cyclohexyl-CH2), 1.30 (1H, d, J= 11.2 Hz, cyclohexyl-CH2), 1.23 (2H,
q, J = 11.6 Hz, cyclohexyl-CH2), 1.31 (2H, q, J = 11.6 Hz,
cyclohexyl-CH2), 1.78-1.81 (4H, m, cyclohexyl-CH2), 1.90 (2H, d, J=
12.4 Hz, cyclohexyl-CH2), 3.53 (211, t, J= 6.0 Hz, -NCH2-), 4.85 (1H,
br, -NH-), 7.40 (1H, t, J= 7.2 Hz, Ar-H), 7.51 (1H, d, J = 8.8 Hz,
Ar-H), 7.55 (1H, d, J= 1.6 Hz, Ar-H), 7.60 (1H, t, J= 8.0 Hz, Ar-H),
7.79 (1H, d, J= 8.0 Hz, Ar-H), 8.50 (1H, d, J= 1.2 Hz, Ar-H), 9.41
(1H, d, J = 8.4 Hz, Ar-H). 13C NMR (100 MHz, CDC13): 6 (ppm)
25.97, 26.52, 31.23, 37.63, 48.50, 120.59, 123.57, 124.41, 125.80,
127.10, 127.38, 129.26, 129.38, 129.45, 130.95, 132.33, 132.41,
134.02, 145.58, 150.64, 180.87 (CO). HRMS (ESI) m/z calcd for
C23H2IN20C1 [M]: 408.1063; found [M+H]: 409.1115.
Example 45=
6-(Benzylamino)-10-chloro-12H-thiochromeno[2,3-dquinolin-
12-one (N25)
Product N25 was prepared from 3 and benzylamine. The pure
compound was obtained as a brown solid (yield 93%) (Rf = 0.67 at
CH2C12: n-hexane = 2: 1). Mp 194-195 C (Me0H). 1H NMR (400
MHz, CDC13): 6 (ppm) 4.94 (211, d, J= 5.2 Hz, -CH2-), 5.16 (1H, br,
-NH-), 7.33-7.51 (611, m, Ar-H), 7.58-7.67 (3H, m, Ar-H), 7.87 (1H, d,
J= 8.0 Hz, Ar-H), 8.59 (1H, d, J = 2.0 Hz, Ar-H), 9.47 (1H, d, J= 8.0
Hz, Ar-H). 13C NMR (100 MHz, CDC13): 6 (ppm) 37.63, 120.98,
123.54, 124.89, 125.87, 127.26, 127.50, 127.63, 128.24, 128.79,
129.43, 129.57, 129.85, 131.02, 132.58, 134.18, 138.82, 145.48,
150.33, 181.00 (CO). HRMS (ESI) m/z calcd for C23H15N20SC1 [M]+:
47
CA 02866502 2014-10-03
. .
402.0594; found [M+Hr: 403.0692.
Example 46
10-Chloro-6-((pyridin-2-ylmethyl)amino)-12H-thiochromeno[2,
3-clquinolin-12-one (N26)
Product N26 was prepared from 3 and 2-picolylamine. The pure
compound was obtained as a brown solid (yield 93%) (Rf = 0.25 at
EA). Mp 187-189 C (Me0H). 1H NMR (400 MHz, CDC13): 8 (ppm)
5.01 (2H, d, J= 4.0 Hz, -CH2-), 6.79 (1H, br, -NH-), 7.24-7.28 (1H, m,
Ar'-H), 7.45 (2H, t, J = 7.2 Hz, Ar'-H & Ar-H), 7.61-7.67 (3H, m,
Ar-H), 7.73 (1H, td, J= 7.6, 1.6 Hz, Ar-H), 7.86 (1H, d, J= 8.4 Hz,
Ar-H), 8.58 (1H, d, J= 2.0 Hz, Ar-H), 8.67 (1H, d, J= 4.8 Hz, Ar'-H),
9.47 (1H, d, J= 8.4 Hz, Ar-H). 13C NMR (100 MHz, CDC13): 8 (ppm)
46.74, 120.81, 122.37, 124.19, 124.61, 125.93, 127.04, 127.60, 129.33,
129.44, 129.60, 131.40, 132.46, 132.52, 134.03, 136.94, 145.62,
148.94, 150.43, 156.58, 181.00 (CO). HRMS (ESI) m/z calcd for
C22H14N30SC1 [Mr: 403.0546; found [M+Hr: 404.0615.
Example 47
64(Benzo[d][1,31dioxo1-5-ylmethyl)amino)-10-chloro-12H-thio
chromeno[2,3-chuinolin-12-one (N27)
Product N27 was prepared from 3 and piperonylamine. The pure
compound was obtained as a brown solid (yield 90%) (Rf = 0.88 at
EA). Mp 205-206 C (Me0H). 1H NMR (400 MHz, CDC13): 8 (ppm)
4.82 (2H, t, J = 5.2 Hz, -NCH2-), 5.08 (1H, br, -NH-), 5.97 (2H, s,
-OCH20-), 6.82 (1H, d, J= 8.0 Hz, Ar'-H), 6.96 (1H, d, J= 8.0 Hz,
Ar'-H), 7.00 (1H, d, J= 1.2 Hz, Ar'-H), 7.47 (1H, td, J= 8.0, 1.2 Hz,
Ar-H), 7.57 (1H, d, J= 8.8 Hz, Ar-H), 7.60-7.66 (211, m, Ar-H), 7.86
(111, d, J= 8.0 Hz, Ar-H), 8.62 (1H, d, J= 2.0 Hz, Ar-H), 9.46 (1H, d,
48
CA 02866502 2014-10-03
. ,
J = 8.8 Hz, Ar-H). 13C NMR (100 MHz, CDC13): 6 (ppm) 46.30,
101.10, 108.40, 108.85, 120.95, 121.58, 123.52, 124.88, 125.86,
127.23, 127.46, 129.39, 129.56, 129.76, 130.98, 132.48, 132.55,
132.61, 134.15, 145.42, 147.07, 147.92, 150.21, 180.93 (CO). HRMS
5 (ESI) m/z
calcd for C24H15N203SC1 [Mr: 446.0492; found [M+Hr:
447.0586, [M-Hf: 445.0440.
Example 48
10-Chloro-642-methoxybenzyl)amino)-12H-thiochromeno12,3
-chuinolin-12-one (N28)
10 Product N28
was prepared from 3 and 2-methoxybenzylamine.
The pure compound was obtained as a brown solid (yield 82%) (Rf =
0.65 at CH2C12: n-hexane = 2: 1). Mp 223-224 C (Me0H). 1H NMR
(400 MHz, CDC13): 6 (ppm) 3.95 (3H, s, -OCH3), 4.93 (2H, d, J= 5.6
Hz, -NCHr), 5.57 (1H, t, J= 5.6 Hz, -NH-), 5.97 (2H, s, -OCH20-),
15 6.94-7.00 (2H,
m, Ar'-H), 7.30 (1H, td, J= 8.0, 2.0 Hz, Ar'-H), 7.45
(1H, td, J = 8.0, 1.6 Hz, Ar'-H), 7.51 (1H, d, J = 7.2 Hz, Ar'-H),
7.59-7.66 (3H, m, Ar-H), 7.89 (1H, dd, J= 8.4, 1.2 Hz, Ar-H), 8.57
(1H, dd,J= 2.0, 0.8 Hz, Ar-H), 9.45 (1H, dd,J= 8.4, 0.8 Hz, Ar-H).
13C NMR (100 MHz, CDC13): 6 (ppm) 42.33, 55.49, 110.47, 120.68,
20 120.81, 123.90, 124.59, 125.82, 126.68, 127.20, 127.52, 128.88,
129.38, 129.44, 129.71, 130.49, 131.23, 132.46, 132.54, 134.04,
145.59, 150.62, 157.89, 181.04 (CO). HRMS (ESI) m/z calcd for
C24Hi7N202SC1 [Mr: 432.0699; found [M+Hr: 433.0783.
Example 49
25 10-Chloro-
643,4-dimethoxybenzyl)amino)-12H-thiochromeno [
2,3-clquinolin-12-one (N29)
Product N29 was prepared from 3 and
49
CA 02866502 2014-10-03
3,4-dimethoxybenzylamine. The pure compound was obtained as a
brown solid (yield 84%) (Rf = 0.66 at CH2C12: n-hexane = 2: 1). Mp
251-252 C (Me0H). 1H NMR (400 MHz, CDC13): 6 (ppm) 3.89 (3H,
s, -OCH3), 3.90 (3H, s, -OCH3), 4.86 (2H, d, J = 4.8 Hz, -NCH2-),
5.11 (1H, t, J= 5.2 Hz, -NH-), 6.89 (1H, d, J= 8.0 Hz, Ar'-H), 7.05
(111, dd, J= 8.0, 2.0 Hz, Ar'-H), 7.08 (1H, d, J= 2.0 Hz, Ar'-H), 7.48
(1H, td, J= 7.6, 1.2 Hz, Ar-H), 7.60 (1H, dd, J= 8.4, 0.4 Hz, Ar-H),
7.65 (1H, dd, J = 8.4, 1.5 Hz, Ar-H), 7.66 (1H, td, J = 8.0, 1.2 Hz,
Ar-H), 7.88 (1H, dd, J= 8.4, 0.8 Hz, Ar-H), 8.59 (1H, dd, J= 1.5, 0.4
Hz, Ar-H), 9.48 (1H, dd, J= 8.4, 1.2 Hz, Ar-H). 13C NMR (100 MHz,
CDC13): 6 (ppm) 46.45, 55.96, 55.99, 111.31, 111.82, 120.59, 120.99,
123.56, 124.88, 125.91, 127.20, 127.50, 129.45, 129.60, 129.88,
131.04, 131.34, 132.59, 134.21, 145.52, 148.63, 149.20, 150.34,
181.01 (CO). HRMS (ESI) m/z calcd for C25H19N203SC1 [M]+:
462.0805; found [M+Hr: 463.0900, [M-Hf: 461.0754.
Example 50
10-Chloro-64henethylamino)-12H-thiochromeno12,3-clquinol
in-12-one (N30)
Product N30 was prepared from 3 and phenethylamine. The pure
compound was obtained as a brown solid (yield 94%) (Rf = 0.52 at
CH2C12: n-hexane= 2: 1). Mp 151-152 C (Me0H). 1H NMR (400
MHz, CDC13): 6 (ppm) 3.10 (2H, t, J= 6.8 Hz, -CH2-), 3.98 (2H, q, J
= 6.4 Hz, -NCH2-),4.91 (1H, t, J = 4.8 Hz, -NH-), 7.27-7.39 (511, m,
Ar'-H), 7.45 (1H, t, J= 8.0 Hz, Ar-H), 7.54 (1H, d, J= 8.4 Hz, Ar-H),
7.59 (1H, d, J= 1.2 Hz, Ar-H), 7.63 (1H, t, J = 7.6 Hz, Ar-H), 7.85
(1H, d, J= 8.4 Hz, Ar-H), 8.54 (1H, d, J= 1.6 Hz, Ar-H), 9.44 (1H, d,
J= 8.4 Hz, Ar-H). 13C NMR (100 MHz, CDC13): 6 (ppm) 35.33, 43.52,
CA 02866502 2014-10-03
t N
120.77, 123.66, 124.69, 125.85, 126.56, 127.23, 127.50, 128.73,
128.94, 129.33, 129.48, 129.50, 131.03, 132.54, 134.08, 139.30,
145.57, 150.36, 180.94 (CO). HRMS (ESI) m/z calcd for
C24H17N20SC1 [M]: 416.9226; found [M+H]: 417.0857, [M+H+2]:
419.0834.
Example 51
10-Chloro-644-methoxyphenethyl)amino)-12H-thiochromeno[
2,3-clquinolin-12-one (N31)
Product N31 was prepared from 3 and
2-(4-methoxyphenyl)ethylamine. The pure compound was obtained as
a yellow solid (yield 95%) (Rf = 0.89 at CH2C12). Mp 214-215 C
(Me0H). 1H NMR (400 MHz, CDC13): 6 (ppm) 3.03 (2H, t, J= 6.8 Hz,
-CH2-), 3.81 (3H, s, -OCH3), 3.94 (2H, q, J = 6.4 Hz, -NCH2-), 4.90
(1H, t, J= 4.8 Hz, -NH-), 6.90 (2H, d, J= 8.4 Hz, Ar'-H), 7.23 (2H, d,
J= 8.4 Hz, Ar'-H), 7.45 (1H, t, J= 7.6 Hz, Ar-H), 7.55 (1H, d, J= 8.8
Hz, Ar-H), 7.59 (1H, d, J = 2.0 Hz, Ar-H), 7.63 (1H, t, J = 7.6 Hz,
Ar-H), 7.85 (1H, d, J= 8.0 Hz, Ar-H), 8.54 (1H, d, J= 2.0 Hz, Ar-H),
9.44 (1H, d, J= 8.8 Hz, Ar-H). 13C NMR (100 MHz, CDC13): 6 (ppm)
34.39, 43.68, 55.31, 114.13, 120.76, 123.67, 124.67, 125.85, 127.22,
127.52, 129.36, 129.49, 129.68, 129.87, 131.05, 131.24, 132.48,
134.09, 145.59, 150.41, 158.31, 180.99 (CO). HRMS (ESI) m/z calcd
for C251119N202SC1 [M]: 446.0856; found [M+Hr: 447.0938.
Example 52
6((4-Aminophenethyl)amino)-1O-chloro-12H-thiochromeno[2,
3-clquinolin-12-one (N32)
Product N32 was prepared from 3 and
2-(4-aminophenypethylamine. The pure compound was obtained as a
51
CA 02866502 2014-10-03
, ,
yellow solid (yield 82%) (Rf = 0.52 at CH2C12). Mp 208-210 C
(Me0H). 1H NMR (400 MHz, CDC13): 6 (ppm) 2.97 (2H, t, J= 6.8 Hz,
-CH2-), 3.63 (2H, br, -NH2), 3.91 (2H, q, J = 6.4 Hz, -NCH2-),4.91
(1H, t, J= 4.8 Hz, -NH-), 6.70 (2H, d, J= 8.4 Hz, Ar'-H), 7.09 (211, d,
5 J= 8.0 Hz, Ar'-H), 7.44 (1H, t, J= 7.6 Hz, Ar-H), 7.55 (1H, d, J¨ 8.0
Hz, Ar-H), 7.60 (111, d, J = 8.4 Hz, Ar-H), 7.63 (1H, t, J = 7.6 Hz,
Ar-H), 7.84 (1H, d, J= 8.0 Hz, Ar-H), 8.54 (1H, d, J= 2.0 Hz, Ar-H),
9.44 (1H, d, J= 8.4 Hz, Ar-H). 13C NMR (100 MHz, CDC13): 6 (ppm)
34.37, 43.70, 115.52, 120.72, 123.73, 124.59, 125.83, 127.19, 127.51,
129.06, 129.31, 129.45, 129.59, 129.75, 131.09, 132.42, 134.04,
144.89, 145.60, 150.45, 180.96 (CO). HRMS (ESI) m/z calcd for
C24Hi6N3OSC1 [M]: 431.0859; found [M+H]: 432.0950.
Example 53
2-(10-Chloro-12-oxo-12H-thiochromeno[2,3-e]quinolin-6-yOgu
15 anidine (TC-SC1-A-41) (N33)
Product N33 was a yellow solid (yield 85%). Mp: 370 C.(dec.)
1H NMR (400 MHz, DMSO-d6): 6 ppm. 7.40 (3H, td, J= 8.4, 1.2 Hz,
Ar-H & -NH2), 7.59 (1H, td, J= 8.7, 1.2 Hz, Ar-H), 7.59 (111, dd, J-
8.4, 0.8 Hz, Ar-H), 7.83 (1H, dd, J= 8.4, 2.0 Hz, Ar-H), 7.95 (1H, d,J
20 = 8.8 Hz, Ar-H), 8.40 (1H, d, J= 2.4 Hz, Ar-H), 9.49 (1H, dd, J= 8.4,
0.8 Hz, Ar-H). 13C NMR (100 MHz, DMSO-d6): 6 ppm. 120.77,
124.49,125.86, 126.82, 128.11, 128.60, 129.21, 129.85, 132.13,
132.52, 132.68, 136.17, 136.80, 144.49, 159.19, 181.16. HRMS (ESI)
calcd for Ci7Hi1N4OSC1 [M] 354.0342; found [M+11] 355.0438.
25 Example 54
10-Chloro-64iperidin-1-ylamino)-12H-thiochromeno[2,3-chu
inolin-12-one (TC-SC1-A-26) (N34)
52
CA 02866502 2014-10-03
Product N34 was a yellow solid (yield 60%). Mp: 180-181 C. 111
NMR (400 MHz, CDC13): 6 ppm. 1.72-1.74 (2H, m, -CH2-), 1.89 (4H,
quin, J= 5.2 Hz, -CH2-), 3.32 (4H, J= 4.8 Hz, -CH2-), 7.36(1H, tt, J-
8.7, 2.1 Hz, Ar-H10), 7.47 (1H, dd, J= 8.4, 2.7 Hz, Ar-H8), 7.61-7.73
(3H, m, Ar-H), 8.00 (1H, d, J= 8.0 Hz, Ar-H), 8.59 (1H,d, J= 2.0 Hz,
Ar-H), 9.63 (1H, d, J= 8.8 Hz, Ar-H). 13C NMR (100 MHz, CDC13): 6
ppm. 24.28, 25.91, 52.36, 123.49, 125.77, 127.32, 127.90, 128.59,
129.06, 129.27, 130.51, 131.66, 132.18, 132.39, 133.49, 134.70,
144.98, 158.53, 181.58.
Pharmacological activity assay
In pharmacological tests, compounds synthesized chemically
including 2-21, N-1 to N-34 (a total of 54 drugs) are subjected to the
following pharmacological activity tests: (1) MTT assay; (2)
Topoisomerase I and II activities assay; (3) cytotoxicity assays
conducted by NCI on the 26 screened compounds in 60 cancer cell
lines.
Example 55
MTT assay for cell cytotoxicity
All of synthesis compounds were evaluated cell cytotoxicity by
using MTT colorimetric assay on PC-3 and DU-145 cell lines.
DU-145 and PC-3 are human hormone-refractory
(androgen-independent) prostatic cancer cell lines from American
Type Culture Collection (HTB-81Tm, ATCC, Rockville, MD)125 and
Bioresource Collection and Research Center (60122, BCRC,
Taiwan)I26, respectively. Two of the "classical" cell lines were
cultivated in RPMI-1640 medium supplemented with 5% fetal bovine
53
CA 02866502 2014-10-03
. .
. .
serum (v/v), 100 U/mL penicillin, and 50 mg/mL streptomycin.
Approximately 2 x 103 cells were seeded into each well of a 96-well
plate and incubated in 5% CO2 at 37 C for 24 h. To evaluate the in
vitro cytotoxicity, all the synthetic compounds were dissolved in
5 DMSO, prepared immediately before the experiments and diluted into
the complete medium before being added to each well of a 96-well
plate. Each compound was then added to the culture medium for
designated various concentrations (0.15, 0.5, 1.5, 5, 15 iM). After 72
h, an amount of 100 ilL of MTT (1 mg/mL) was added to each well,
10 and the samples were incubated at 37 C for 4 h. After removing the
MTT solution, 100 ilL of DMSO was added to each well and
incubated at 37 C for another 20 mins. The absorbency at 560 nm
was measured by using an ELISA reader.
Results are expressed as mean values of at least three
15 independent experiments. The IC50 values of testing compounds were
described in Table 1.
Tablet . Effects of 1 0-Chloro- 1 2H-thiochromeno [2 ,3 -c] quino lin
-12-one derivatives on cytotoxicity by MTT assay.
MTT assay (IC50 SD) a
No. R substitutions
DU-145 (j1M) PC-3 (iaM)
0 OH
2 S
0 > 15
N OH
3 1¨CI >15 >15
4 I-0H >15 10.10
1.81
I¨OCH3 10.84 6.55 3.89 0.54
6 1---N/ >15 >15
\
54
CA 02866502 2014-10-03
, .
7 i-N/-S
NH 5.01 1.68 2.84 0.64
8 i-N/--\N- 12.94 0.26 7.18 2.45
\/
1-N N i--\ /
9 >15 12.04 3.41
OH
i-N/---\N -I- >15 >15
11 /--\ 41 >15 >15
I-N N
1r-\2 I-N N 4. >15 >15
/ /----\
13 LN 0 >15 >15
//---\
14 rN S >15 >15
I-N ) 13.38 2.87 >15
16 I-N )-OH 11.12 4.18 9.55 2.42
17 . >15 >15
1--N
/ /
18 1-N )-N/ ) 10.23 2.03 14.16
1.41
\ \
AN ----." NH
19 9.02 1.20 6.36 0.17
/ /-----
>15 >15
r-N\--
M
N 0
21 e" ---- 11.12 4.18 9.55 2.42
--,...----
N1 FNHcH3 11.65 5.15 14.48
2.70
N2 /N '= 6.70 1.62 5.18 2.20
H
N3 A N'. 7.64 2.74 7.41 3.43
H
N4 A N ..--,,,,,,,, 10.35 2.46 10.11
1.32
H
N5 A N .-,,,,,,,,, >15 >15
H
CA 02866502 2014-10-03
N6 AN >15 >15
N7 N 5.94 3.45 2.25 0.61
N8 1.73 0.72 1.11 0.69
N9NOH 11.55 7.23 7.48 3.35
N10'N-OH 5.94 2.03 6.31 4.11
N11 N 4.95 0.58 8.38 3.39
N12 NOH 6.74 2.14 7.05 1 2.48
N13 A N 14.16 1.18 10.38 2.30
N14 NNH2 1.80 0.40 1.64 0.55
N15 A
N N'OH 3.81 1.88 3.44 1.81
ro
N16 AN,Nj >15 >15
N17 /Nre 2.70 0.16 2.27 0.09
N18 AHNINC 3.53 1.05 2.22 1 0.21
N19 A
1.50 1.32 1.98 0.72
N20 4 4.86 0.99 6.20 2.52
N
N21 ANJO 8.06 2.08 7.51 0.22
N
N22 AN 8.39 1.84 6.47 0.30
N23ANS\, 14.65 3.84 >15
H
56
CA 02866502 2014-10-03
. .
. .
Am
N24 j_i 13.55 3.88
14.80 3.26
AN
N25 j.1 0 13.49 0.69
11.16 1.55
ii.,....--..,...N,,
N26r i i ` 14.75 1.99
14.20 2.22
N27 H
AN 0 > 11.82 5.83 >15
0
AN
N28 H 1.1 9.61 4.88 6.01
3.31
H3C0
N29 AN 0 C) 4.86 0.99 6.20
2.52
0
N30 AN 10 >15 >15
H
0 OCH3
N31 i >15 >15
TN
H
0 NH2
N32 ,4 13.59 1.39
10.11 0.94
1 -N
H
NH2
N33i ,..1,, 8.11 1.37 6.13
6.98
r- N NH2
-1-
cht NH
N34 9.06 1.95 8.22
1.02
- Mitoxantrone 0.10
0.01 0.39 0.02
- Doxorubicin 0.12
0.03 0.63 0.26
- Camptothecin 0.10
0.01 0.10 0.01
- Etoposide (VP-16)
0.40 0.01 4.33 0.86
a SD: standard derivatives, all experiments were independently
performed at least three times.
Besides, compounds N7, N8, N14, N15, N17, and N18
containing more than one nitrogen atom in the side chains showed the
outstanding cytotoxic activities than having a hydroxyl group, alkyl
57
CA 02866502 2014-10-03
group, or aromatic rings. Compounds 5, 7, 8, 16, 19, N2, N7, N8, N9,
N14, N15, N16, N17, N18, N19, and N25 were selected for TOPs
activities assay.
Example 56
Topoisomerase I and II activities assay
According to the cell cytotoxicity, compounds 5, 7, 8, 16, 19, N2,
N7, N8, N9, N14-N19, and N25 were also selected for primary TOP I
and II activities assays at 25 and/or 50 M (Fig. 2-4). In TOP I activity
assay, compounds 7, N7, N14, N15, N17, N18, and N25 were showed
more potent inhibitory effects than CPT and selected for further
evaluation by using five different concentrations (Figure 4). The ICso
value of compounds 7, N7, N14, N15, N18, N19, and N25 were about
10, 10, 1, 5, 25, 5, and 25 M, respectively (detect by TopoGEN
TG2005H, TG-2000H-1).
We also performed TOP II-catalyzed relaxation of plasmid DNA
assays(Fig. 5-7) to evaluate if compounds could inhibit TOP II.
Respond to our drug design, compounds 7, N7, N8, N14, N15, N18,
and N19 were showed more potent inhibitory effects than the positive
agent VP-16 and selected for further evaluation by using five different
concentrations (Figure 7). The IC50 value of compounds 7, N7, N8,
N14, N15, N18, and N19 were about 10, 10, 1, 10, 5, 1, and 1 M,
respectively (detect by TopoGEN TG2005H, TG-2000H-1).
Example 57
National Cancer Institute Cancer Cell Cytotoxicity assay:
The test results shown in this section are the compound
58
CA 02866502 2014-10-03
cytotoxicities in vitro against cancer cell lines National Cancer
Institute (NCI)'s anticancer drug screen and 26 compounds (2, 3, 4, 5,
6, 8, 10, 11, 12, 13, N1, N2, N6, N7, N9, N12, N13, N14, N16, N17,
N19, N21, N25, N27, N30, N31) screened. In the first stage,
cytotoxicity of the 26 compounds at the concentration of 10 p,M was
conducted on 60 cell lines and SRB assay was performed after 48
hours of incubation. The results are shown in Tables 2 to 4 and are
represented by growth percentage.
Furthermore, among five compounds were active drugs for
further their cytostatic and cytotoxic activities against the 60 cell panel
by using five dose studies (0.01, 0.1, 1, 10 and 100 p,M) (Table 5).
Many changes and modifications in the above described
embodiment of the invention can, of course, be carried out without
departing from the scope thereof. Accordingly, to promote the
progress in science and the useful arts, the invention is disclosed and
is intended to be limited only by the scope of the appended claims.
25
59
CA 02866502 2014-10-03
. .
. .
Table 2. In-vitro anticancer activity of compounds 2, 3, 4, 5, 6, 8,
10, 11, 12, and 13 in NCI's drug screen program.
Compounds/ Growth percent'
2 3 4 5 6 8 10 11 12
13
Panel/Cell line L:-, F. F. F. g V ! . "e 1
:2
r- ao
,
t... r,
. 72 1----. ,
P fz' r=-:. Fs. r,
..,.- .
r
i-, t; ,
cd ,.) t:, 4.),
U td-- ,.
cn cn cn w cn cn V) V)
co cn
Z Z Z X Z Z X Z Z
Z
Willialiiii,..:i:77;POIRW:' - is'ilinliAlsell 46,:ifittv4ici it . ' :MN Ili Ri
-*pi ., = I lifiamtperetiii .!.. .44:.f-':-..!
CCRF-CEM 98.31 93.12 107.78 106.56 99.42
91.60 83.46 95.42 98.41 88.02
HL-60(TB) 105.63 99.34 109.82 89.47 107.16
96.19 9311 100.96 113 79 93.38
K-562 112.29 101.04 107.08 95R3 99.71
72.98 35.47 87.84 112.37 81.18
MOLT-4 97.25 98.04 100.41 80.62 97.52
87.97 7001 90.91 107.23 89.31
RPMI-8226 98.10 100.19 100.99 102.86 98.20
91.11 84.87 96.63 99.93 89.54
SR 106.33 8Q02 96 00 88.18 95.68
86.41 79.07 .5.1 103.11 88.91
044414081811111el4liO17W i'V..-' ::'1! ' F..... iig: F,' ':;' = II i'
ilWaralaWa:W., ;Ka. SiCi 1:12iiillt . ' k = RitfilElk ::1441114
A549/ATCC 107.34 104.74 95.25 100.46 94.16
99.80 95.49 106.37 98.54 92.97
EKVX 89.47 N.T. N.T. N.T. N.T. N.T.
N.T. N.T. N.T N.T.
HOP-62 116.47 82.02 9080 69.64 43.62
49.41 67.83 87.72 86.00 70.77
HOP-92 72.97 90.83 93.13 80.11 58.65
17.97 39.64 70.17 81.27 60.43
NCI-H226 105.64 83.41 99.58 98.57 96.20
85.67 89.29 88.98 87.10 88.01
NCI-H23 92.61 85.12 88.96 80.14 77.00
81.11 84.75 89.94 93.80 95.21
NCI-11322M 101.10 100.12 90.87 90.77 83.74
77.04 82.85 88.92 84.41 88.41
NCI-H460 104.20 103.14 96.92 101.23 88.86
96.05 93.71 95.57 103.24 9928
NCI-H522 81.64 96.81 86 60 92.71 70.42
98.78 94.76 96 69 93.50 89.60
FilhiliAlligialigninallithiet E=,==,:i2-,:j: ..tilitg47;t11501418111, iiN:::'
It itljil In : ::!!' gUilaillitailiglial :,111,4i.11.1414
COLO 205 108.65 112 15 93.69 97.94 89.67
9006 88.90 105.64 111.67 99.24
HCC-2998 100.97 123.28 101.48 104.67 97.20
96.87 103.49 99.32 105.23 108.75
HCT-1 16 103.87 10/27 86.20 89.48 74.26
80.94 69.33 98.59 105.36 85.71
HCT-15 107.80 102.28 98.1 1 107.99 89.27
93.94 8/92 94.51 99.81 93.81
HT29 101.41 99.73 96.78 102.69 87.73
78.65 63.19 102.71 99.94 83.73
KM12 112.61 111.27 102.45 98.03 96.40
110.71 101.86 107.75 113.15 88.68
0.W-670 0,92 99.06 101.39 103.05 105.18
95.50 88.11 93 08 100.20 96.18
ilittLviiiii , ' .--411: .,, '-= iiM,.LOIR::; .:zigiaii....qiii:
HantuoirlioN.vtglivanwlincy,a =::-,H ..,.1tti =1:-.g0
SF-2os 113.06 109.60 95.26 94.12 77.86
82.98 87.80 100.42 103.82 103.56
SF-295 98.99 90.26 99.93 86.35 48.82
91.69 100.32 94.34 9923 93.62
SF-539 110.22 89.02 93.21 86.13 72.72
85.30 86.18 108.56 100.06 82.38
SNB-19 97.17 107.89 101 37 89.36 76.46
87 07 84.81 95.52 105.57 9937
SNB-75 82.18 77.88 66.63 52.28 15.29
42.15 77.64 8/62 94.73 76.88
U251 99.50 101 71 101 85 99.18 93.55
92.52 83.31 96.69 09 99
WarailinV14.59111f ' ;', :q.7.3V_ = .44faiL:=!:: 117ii ' e , :
: . 98r.5i8
i r ol
l
LOX IMM 00.46 90.53 92.07 91.21 8653
87.47 87.79 94.24 95.15 89.82
MALME-3M 107.05 95.87 93.26 94.19 89.70
108.36 119.20 112.36 121.84 118.84
M14 106.90 106.18 94.76 88.40 94.36
102.15 101.70 93.00 104.02 110.35
MDA-MB-435 102.02 100.64 103.18 103.83 107.18
105.80 98.24 96.99 102.39 102.07
SK-MEL-2 119.91 95.54 103.04 105.52 87.26
97.53 104.06 107.37 104.26 98.77
SK-MEL-28 98.95 101.80 116.59 112.31 96.60
107.92 100.66 104.19 104.22 111.28
SK-MEL-5 98.07 97.42 100.69 99.74 97.92
99.54 94.13 98.38 103.57 95.88
UACC-257 121.83 104.47 108.36 108.09 109.22
100.55 102.79 95.39 93.68 110.91
UACC-62 102 9; 89 60 105 06 96.15 87.80
99.27 91.36 96.00 104.48 87.86
Wititinliiiiii AEC ., = ',:- ' , . -i.,11 , ' .:'. h: . 1111:!. ' T'''. r
.:1:.:7,i1E,.}NiNtiliniir 1..i,I.:WW ,11r0 ;:i4VitiCtRI
IGROVI 103.57 96.06 7835 71.44 68.16
80.08 71.67 93.49 91.38 80.90
OVCAR-3 121.29 11 1.27 106.37 96.60 79.65
89.13 93.92 101.69 10963 96.54
OVCAR-4 98.41 87.91 N.T. N.T. N.T
94.93 85.35 102.43 94.29 96.75
OVCAR-5 11/47 100.07 111.17 106.70 98.05
98.14 95.70 111.92 108.10 95.97
OVCAR-8 114.68 101.62 100.55 91.82 68.53
89.44 84.95 98.54 90.34 83.66
NCl/ADR-RES 97.75 102.80 96.84 96.10 87.86
81.35 92.05 98.31 100.81 98.16
SK-OV-3 N Tb 101.32 .. 44.40 58.94 41.57
75.11 95.12 . . 94.79 100.39 77.18
ratifialitai-44 iiiingr ' '44f-- - ': ''''- "':-.::.i4liiii411.''.'7.ir': I.
i'i'. SPY/AMU:1- 0.. -,510.1t Eli Ttigatirktfrag,',-tliVi -.1E4 = flu
786-0 104.44 405.25 99.76 99.15 87.00
90.48 85.58 105.57 101.58 99.31
A498 N.T. 106.06 107.74 96.29 97.06
79.33 62.20 94.17 81.04 94.15
ACHN 113.47 86.84 95.52 74.89 64.81
89.73 83.05 96.39 94.99 82.54
CAKI-1 76.26 94.31 NT N.T NT 87.50
87.87 94.03 97.83 90.00
RXF 393 113.90 102.55 116.94 109.68 83.17
88.57 70.31 100.71 107.68 108.61
SNI/C 95.83 95.92 101.14 98.74 99.49
83,03 86.81 97.70 102.66 96.81
TK-10 118.09 108.56 113.26 113.30 64.75
97.07 93.19 124.99 112.45 86.24
CO-31 68.91 66.06 "6.44 74 F. 73 81
48.97 42.34 64.91 V 45 73.11
igsgereinammgraggint -.. '.: :i , ,'. -
, + ,- = '. , : ' ir . : P. MIN1177,11; I - '--,Ivi.-.5.71i-f?.,T;p51:?
4:41p5mirg
PC-3 93.23 91.59 .2 4 -I I .4 ,
84.81 77 9., 79.80
DU-145 115.22 114.27 , , .
' =.i , I 103.45 962, ;II , I II, , 110.5'1
fatininger*-gt114111111/1144M-r.t..1.' Ti'.k 1- '_ '1 ' .f.'" 4104 ti!
T.T',',: ,,11,1,-, ' , -:. .'' -', = = ' :: NvliZIFI
MCI,/ 91.95 82.38 69.90 65.39 22.19
76.60 51.73 68.32 74.16 50.59
MDA-MB-231/ATCC 95.70 84.28 81.80 78.12 71.03
57.30 55.49 89.11 94.34 71.53
HS 5781 88.33 95.14 113.28 66.51 60.38
65.57 76.01 76.42 85.20 83.27
BT-549 124.20 110.01 85.17 94.85 92.96
96.25 90.05 114.05 123.95 113.81
T-4713 99.59 110.10 77.89 68.46 37.66
82.24 87.22 89.22 104.57 80.00
MDA-MB-486 104.57 98.57 100.33 103.54 29.91
102.02 81.67 98.53 86.34 39.67
Mean 102.11 98.11 97.17 92.10 80.59
86.81 84.06 96.19 9930 90.52
Delta 33.20 32.05 30.54 39.82 65.30
6864 48.59 31.28 35.45 50.85
Range 55.29 57.22 50.31 61.02 93.93
92.74 83.73 60.08 60.10 79.17
a Data obtained from NCI in vitro 60-cell drug screen program at 10-5
molar concentration. b N.T. = No test.
CA 02866502 2014-10-03
.., .
Table 3. Growth percentage of compounds N1, N2, N6, N7, N9,
N12, N13, and N14 in the NCI in vitro 60-cell Drug Screen Program.
Compounds/ Growth percent'
NI N2 N6 N7 N9 N12 NI3
N14
:2
Panel/Cell line 4, -
r- F-- 'Fn-
1--
in cn z in in
Z Z Z Z 4 4
4
Leukemia ______________________ ..
CCRF-CEIS4 79.77 88.62 93.72 -53.54 81.07 70.00 98.57
-3961
HL-60(TB) 131.66 103.33 141.93 -47.40 104.58 96.29
103.96 -47.50
K-562 40.69 77.52 95.25 -58,60 68.23
46.86 106.64 -57.17
MOLT-4 86.86 92.27 103.08 -56.34 78.76
61.79 93.68 -58.82
RPMI-8226 93.10 79.66 108.60 -37.68 83.81 73.50 97.78
-27.48
SR 47.76 84 32 104.59 -34.76 60.44
62.38 97.29 -5167
Non-snuill cell lung cancer . . .
A549/ATCC 51.68 61.84 91.04 23.38 80.72 69.86 101.98 -
88.76
EKVX N.T. N.T. NT. N.T. N.T. N.T.
N.T. N.T.
HOP-62 79.39 57.49 95.32 56.42 81.90
55.91 88.69 -97.40
HOP-92 59.15 -0.07 83.26 -10.12 63.17
N.T. N.T. N.T.
NC1-H226 73.53 41.31 79.22 58.08 83.55
77.56 87.55 28.55
NCI-H23 44.39 86.29 94.17 72.59 86.06
81.25 95.10 -5.25
NCI-H322M 60.89 74.88 92.45 49.16 79.00 58.98 85.51 -
50.28
NCI-H460 73.84 53.33 99.42 -71.14 100.97
83.78 95.17 -6239
NCI-H522 97.00 80.75 99.02 -8.99 76.94 64.07 102.30 -
66.59
Colon cancer , = . '
COLO 205 94.82 82.36 107.68 -82.85 99.13 68.37 93,45
-87.22
HCC-2998 105.34 98.02 119.79 -8365 101.55 99.61 112.26
-90.43
HCT-I16 32.49 48.13 99.01 -96.59 93.62
73.42 90,30 -48.19
HCT-15 68.73 79.72 95.35 -30.05 78.61
77.58 96.97 -45.92
HT29 114.79 68.21 11670 -84.24 87.36
78.24 121.02 -83.30
KMI2 68.75 8579 9404 -7707 91.93
8237 105,66 -859
SW-620 84.37 84.65 104.94 -67.82 97.18
84.32 92,80 -66.88 ,
. .
CNS cancer .
.
SF-268 70.58 78.23 102.86 39.05 95.06
76.71 105.52 -67.65
SF-295 89.04 69.49 104.18 -32.59 92.50
89.58 95.92 -74.29
SF-539 59.86 60.35 95.09 48.73 90.56
83.86 N.T. 2.13
SNB-19 96.48 84.19 99.01 55.44 90.96
93.87 106.79 40.06
SNB-75 67.29 12.12 73.24 33.36 90.05
31.01 85.02 -43.13
U251 85.19 76.58 97.05 -39.75 84.10
74.49 105.12 N.T.
Melanoma
. =
LOX IMVI 44.95 80.61 89.35 -84.12 86.96 79.25 98.90
N.T.
MALME-3M 72.46 77.31 92.36 -4.32 104.86 58.66 95.93 -
48.42
M14 105,85 91.72 107.34 -92.77 107.09
85.38 94.27 -55.62
MDA-MB-435 8497 80 67 108.13 -6464 94.98 92.11 108.08 -
85.39
SK-MEL-2 110.10 76.15 104.97 -7.83 86.80 N.T. N.T.
N.T.
SK-MEL-28 83.41 93.07 98.40 -7679 100.76 82.86 106.87 -
96.20
SK-MEL-5 80.13 73.99 90.08 18.83 97.36 87.19 90.71
68.64
UACC-257 91.23 75.21 99.44 -4822 89.33 95.71 114.40 -
67.35
UACC-62 77.66 69.06 83.74 -76.58 76.52
76.17 96.09 -96.67
Ovarian cancer =
1GROV I 67.97 59.26 80.40 33.20 88.51
66.54 74.77 -77.31
OVCAR-3 N.T. 76.51 N.T. - 19.74
103.35 96.34 115.22 -84.15
OVCAR-4 71.33 23.19 98.77 60.67 102.67
83.23 86.07 -20.15
OVCAR-5 86.91 92.93 99.96 24.27 101.56
88.01 101.36 -5216
OVCAR-8 31.61 62.54 95.31 1,22 78.13
68.74 92.79 -74.59
NCl/ADR-RES 70.95 88.61 101 32 23.00 91.54 77.02 98.47
-42.68
SK-OV-3 88.67 2890 99,71 70.10 104.57
58.93 86.70 12.25
õ : -.
= . =
786-0 95.51 23.87 108.84 43.11 10443
82.86 99.74 -57.40
A498 82.89 61.02 103.44 43.17 65,78
74.62 75,01 60.29
ACHN 84.47 43.77 84.92 7.55 82.25
69.12 91.24 -92.31
CAK1-1 68.09 72.44 73.26 34.60 89.29
59.29 86.38 -75.30
RXF 393 84.52 34.56 93.23 -15.29 92.81
71.61 98.67 -87.22
SNI2C 92.52 85.67 94.28 48.67 92.52
81.98 95.41 -93.98
TK-10 81.21 47.11 138.89 -89.05 78.14
77.45 123.23 -28.04
U0-31 65.34 32.06 78.57 21.83 51.97
51.08 78.64 -92.77
= =: Proittateetileee : ,
= = - ..
PC-3 61.41 77.80 84.63 33.32 71.70
58.35 86.33 -97.71
DU-145 80.29 87.67 109.54 -80.85 102.77
87.04 113.48 -96.35
Breast cancer .
MCF7 23.26 17.54 67.12 -85.41 29.66
40.92 73.82 -98.38
MDA-MB-231/ATCC 66.58 4866 77.83 -37.28 66.62 58.29 84.22
-84.52
HS 578T 82.63 36.37 89.81 34.31 83.24
52.66 89.33 50.82
BT-549 108.49 100.10 109.76 91.29
131.62 85.96 103.61 78.49
T-47D 42.13 38.28 73.62 26.06 74.45
46.94 73.76 -82.45
MDA-MB-486 8.75 26/9 95.79 12.81 39.59
80.85 96.02 -62.13
Mean 75.06 65.97 96.94 -1106 86,33
73.52 96.33 -51.89
Delta 66.31 66.04 29.82 85.53 56.67
42.51 22.57 46.49
Range 122.91 103.40 7481 187.88 101.96
68.60 49.47 176.87
aData obtained from NCI in vitro 60-cell drug screen program at 10-5
molar concentration. l'N.T. = No test.
61
CA 02866502 2014-10-03
. .
. .
Table 4. Growth percentage of compounds N16, N17, N19, N21, N25, N27,
,
N30, and N31 in the NCI in vitro 60-cell Drug Screen Program.
Compounds/ Growth percent*
N16 N17 N19 N21 N25 N27 N30
N31
Panel/Cell line
E F
I; Li tz
,..., r
u
. . . . .
.
R z z z z z z
z
. .
. . . .
Leukemia = = .: = -= , . = ,
. .
=
CCRF-CEM 82.21 69.06 -54/8 92.23 102.34
88.86 114.29 100.93
HL-60(TB) 104.97 77.32 -37.88 107.06 98.82
112.12 105.68 94.27
K-562 17.67 47.53 -66.94 102.86 90.61
80.37 82.82 91.76
MOLT-4 80.91 70.71 -47.57 92.42 92.42
87.55 87.33 82.02
RPMI-8226 92.20 78.66 5.53 94.56 88.54
96.35 77.36 99.98
SR 67.74 48.48 -51.40 85.52 85.46
79.57 89.88 94.13
,Non-small cell lung cancer
A549/ATCC 95.12 81.19 -61.45 85.28 68.33
104.74 88.44 93.49
EKVX N.T.' N.T. N.T. N.T N.T. N.T
N.T. NT
HOP-62 67.97 86.73 36.97 101.46 66.53
78.01 103.19 90.07
HOP-92 58.79 40.76 23.01 N.T N.T.
45.39 73.02 67,57
NCI-H226 94.09 81.01 86.75 93,26 79.35
53.50 77.00 72.39
NCI-H23 83.07 100.25 54.26 91.11 74.37
79.93 89.89 92.72
NCI-H322M 76.96 82.29 1005 91.10 91.00
73.65 117.72 99,19
NCI-H460 10049 88.06 -80.98 91.29 64.41
76.71 95.11 98.14
NCI-H522 76.66 98.76 -59.68 83.95 75.71
78.43 93.93 93.69
Colon cancer ._ . =
. .
COLO 205 79.39 91.35 -87.17 103 96 80.79
87.63 100.02 105,41
HCC-2998 90.51 108.36 -74.47 109.83 95.77
N.T. 113.77 105.27
HCT-116 81.64 70.19 -91.19 65.37 66.87
69.98 85.77 9846
IICT- 15 58.81 80.92 -76.47 91.71 71.76
77.41 91.82 100.67 =
HT29 43.13 90.98 -83.53 105.11 77.16
82.91 109.15 12078
KM12 68.16 97.75 -83.29 112.90 80.34
87.96 90,99 109.46
SW-620 99.65 91.74 -81.48 87.98 78.97
76.95 93.45 97.10
.
CNS cancer , ,.
SF-268 79.49 87.67 17.28 92.06 73.68
93.24 101.32 100.14
SF-295 102.17 95.14 -93.00 92.22 74.33
94.39 95.46 95.31
SF-539 76.72 87.85 -47.59 91.20 80,59
77.12 97.86 95.67
SNB-19 90.44 97.83 46.07 106.72 10/93
97.62 100.86 96.74
SNB-75 60.74 60.99 23.66 81.91 14.83
57.14 68.76 74.50
1J251 76.34 84.19 -66.76 NT N.T.
, 81.03 96.78 98.72
' Melanoma = =: =. ,
. ... .. .
LOX IMVI 67.43 66.16 -8239 N.T. N.T.
78.69 89.03 87.78
MALME-3M 78.79 100.91 -3430 93.42 77.59
81.64 122.61 99.74
M14 99.59 103.73 6.12 103.41 87.29
94.93 97.97 100.06
MDA-MB-435 98.93 93.04 -88.95 106.98 89,58
91.64 106.06 104.49
SK-MEL-2 94.52 106.47 53.64 N.T. N.T
95.57 106.33 113.12
SK-MEL-28 98.56 99.63 -94.33 101.79 84.20
88.24 98.94 99.41
SKMEL-5 91.61 91.73 62.02 97.00 92.81
79.86 89.49 85.38
UACC-257 106.52 107.40 45.70 103.68 83.24
N.T. 109.29 98.66
UACC-62 94.58 102.63 -77.63 93.23 89.20
72.96 74.67 80.17
Ovarian cancer . ' : . = ,
IGROV1 74.11 87.27 -43.25 83.31 53.42
67.81 N.T. 86.73
OVCAR-3 97.14 N.T. N.T. 91.85 76.59
N.T. N.T. NT
OVCAR-4 N.T. 94.63 12.32 72.10 27.24
53.73 79.53 79.00
OVCAR-5 68.98 106.26 -65.05 98 06 91.27
84.38 10/03 100.83
OVCAR-8 92.43 98.38 9.89 74 91 68.33
79.45 95.61 97.83
NCl/ADR-RES 77.03 9694 26.35 95.71 77.36
91.11 96.51 103.78
SK-OV-3 85.34 99.91 76.48 94.54 36.71
82.87 102.16 96.11
, . . . .
Rene cancer ' . = : ,
-
786-0 83.16 93.30 -17.60 97.69 72.48
95.83 108.00 100.71
A498 80.54 78.57 61.88 93.00 86.46
75.91 83.22 99.56
ACHN 91.61 86.20 -96.96 97.31 51.82
5848 90.85 91.19
CAKI-1 N.T. 8431 N.T. 72.76 80.38
75.09 81.04 85.75
RXF 393 93.21 76 32 43.67 97.03 71.08
74.20 96.74 88.50
SN12C 95.98 83.63 78.75 89.12 87.82
85.53 86.54 94.23
TK-10 76.67 108,89 -92.61 12796 75.80
116.79 122.13 132.57
.
U0-31 54.16 72.62 24.54 72.52 65.31
80.98 75.82 72.12
.. . ... ,. . , . ,
Prostate cancer '= -= = ' - ==
PC-3 67.82 81.31 -26.98 75.02 72.70
78.54 87.25 84.95
DU-145 93.09 97.29 -71.03 104 /5 74.80
72.44 100.17 107.03
Breast cancer
MCF7 51.69 51.20 -7/49 46.14 1601
43.53 60.91 59.13
MDA-MB-231/ATCC 59.20 73.39 -66.53 77.17 32.67
54.65 79.55 82.31
HS 578T 89.65 87.67 64.69 94.22 34.73
70.49 87.62 87.49
BT-549 91.21 99.81 98.48 105.53 97.19
111.06 106.85 105.24
T-47D 47,63 79.82 16.26 72.43 38.08
74.41 71.96 69.90
MDA-MB-486 74.34 43.26 -24.00 108.92 -9.83
60.47 63.11 53.93
Mean 80.38 85.32 -21.33 92.62 71.97
80.18 93.22 93.38
Delta 62.71 44.56 75.63 46.48 81.80
36.65 32.31 39.45
Range 88.85 68.13 195.44 81,82 112.76
73.26 61.70 78.64
'Data obtained from NCI in vitro 60-cell drug screen program at IV molar
concentration. bN.T. = No test.
62
CA 02866502 2014-10-03
. .
. .
Table 5. In vitro antitumor activity (G150 in M), toxicity (LC50 in
.1\4) and TGI data of selected compounds N2,N7, N14, N19, and N25.
N2 N7 N14 N19
N25
Panel/Cell line
(pM) (NSC771784) (NSC771785) (NSC772865) (NSC777201) (NSC772867)
G150 TGI LC so GIso TGI LCso GIso TGI LCso Glso TGI LCso GIs(
TGI LCso
16416041. -6'1 3,1'4.4;.e.'1,.,,... l'..,"...,...'r.,,-i,,;.:',I
,.:.....i ..'li .: : "..:..., ',!;:::.,.i.ii:,.i.,. 4.:.i: jl.
.i,'''J - i ;. ' = .. :!? 1.1 i:LI0'-!i :' 111:1I :i4. ....i.4.!
eCliF-EM > 100 , 100 > 100 1.63 3.41 7.15
.1.7.5 3.65 7.64 1,83 3.80 > 100 > 100 > 100
HL-60(TB) >100 >100 >100 1.76 3.49 6.95 N.T. N.T. N.T. N.T. N.T.
NT N.T. N.T. NT.
K-562 >100 >100 >100 1.33 3.00 6.74 1.69
3.65 7.84 1.73 >100 >100 >100 >100
MOLT-4 > 100 > 100 > 100 1.71 3.40 6.74
2.02 3.96 7.77 1.88 - > 100 > 100 > 100
RPMI-8226 > 100 > 100 > 100 1.76 3.68 769 2.26
4.92 62.30 1.84 4.21 .. > 100 > 100 > 100
SR ... . , 100 , 100 >. 100 _ L70 . . 3;73 .. 8.14 r
I RI 4.30 = 120 198 1.55 , . > loo . ...> too . . loo. . >100
nitiW4841170thingealkiK' .1.1.;=:ti:% F '.7: -r='.''. :.. : .i, ' l'i ' '..'1
'' i !: : '..!,A - ' :' L = 11! !'' = 1:'= 1,.. :i'.' i:.11. .! : "
;'INIE7. lit : 1:ii.'11 ''.: .: '. = .: ...:.. 71 ,::).iiiitr:!
A549/ATCC . 48.1 > 100 > 100 1.66 3.17 6.06
1.83 3.50 6.68 1.76 3.30 14.40 > 100 > 100
EKVX NT N.T. N.T. N.T. N.T. N.T. N.T.
N.T. N.T. NT. N.T. NT N.T. N.T. N.T.
HOP-62 13.30 35.20 93.10 1.54 2.92
5.56 1.47 2.92 1.38 2.89 2.43 5.92 27.70
HOP-92 N.T N.T. N.T. N.T, NT NT 1.03
3.05 9.04 1.23 4.58 26.70 1.75 7.53 53.60
NCI-H226 33.20 > 100 > 100 1.84 4.38 24.30
1.36 2.95 1.90 4.46 > 100 1.84 > 100
NCI-H23 N.T. N.T. N.T. N.T. N.T. N.T.
1.72 3.37 6.63 1.89 3.68 15.20 > 100 > 100
NCI-H322M 69.50 > 100 > 100 1.74 3.18 5.81
1.65 3.34 6.74 1.80 3.56 7.03 > 100 > 100 > 100
NCI-460 > 100 > 100 > 100 1.76 3.38 6.50
1.69 3.45 7.05 1.66 3.35 5.00 > 100 > 100
NCI-H522 24.80 > 100 > 100 1.59 3.07 5.90
1 RA 3.59 6.92 1.85 3,56 . 7, 25.30 . > /00, > 100
111.10-4",f,,,.4.i4iii,11,
COLO 205 > 100 > 100 > 100 1.66 3.17
6.05 1.73 3.57 1.63 3.37 > 100 > 100
HCC-2998 > 100 > 100 > 100 1.68 3.38 6.82
2.03 4.06 8.14 1.58 3.42 - > 100 > 100 > 100
HCT-116 6.31 > 100 > 100 1.57 3.18 645
1.63 2.99 5.47 1.60 3.05 5.59 > 100 > 100
HCT-15 > 100 > 100 > 100 1.59 3.14 6.21
1.58 3.12 6.16 1.61 3.25 6.57 8.75 > 100 > 100
HT29 > 100 > 100 > 100 1.65 3.36 688
1.78 3.29 6.09 2.05 4.00 7.82 > 100 > 100
KMI2 > 100 > 100 > 100 1.78 3.28 6.03
1.67 3.11 5.79 1.67 3.31 48.30 > 100 > 100
SW-620 > 100 > 100 > 100 1,71 3 79 6 36
1.67 3 42 7 07 1 61 1 ln _ >100 >100
.rli. . .- - -it .:.-; ,;.': ,' _'-j'. ::'
.1-' = f- .7 i-__.i.... ' .'-'_',,f'., _ -. ithitilig,' --
:!: ., .- . ---=,-11,
SF-268 22.10 > 100 > 100 1.57 3.16 6.35
1.59 3.23 6.55 1.67 3.35 - 13.80 > 100 > 100
SF-295 12.20 35.20 > 100 1.73 3.21
5.98 1.79 3.63 1.73 3.33 3.50 16.50 > 100
SF-539 10.10 58.50 > 100 1.68 3.08 5.67
1.57 2.95 5.52 1.70 3.22 6.11 4.49 46.40 > 100
SNB- I 9 2660 81.10 > 100 1.54 3.07
6.10 1.65 3.18 1.64 3.26 8.56 > 100 > 100
SNB-75 4.96 22.00 60.20 1.28 3.69 12.10
1.09 2.31 4.90 1.28 2.56 5.14 1.45 3.78 9.91
U251 16.90 67 90 > 100 1.48_ ._.
214._ 5.44 r rrr. ,r 1.63 ..r r 3.07 r; 5.78. . . r 1,73 ,...... 3.32 _
,_,4.4t. 2..1.0>.:109 .,
1:;1:-!.E
.:l..V.i!.$..1.44 "III, '',.!'"fiT. Ict-i71..' '.-'1!! ilill'PE
3,4i.'..13)
Lox imvi - >100 >100 >100 ..ob 3.24 6.24
1.68 3.16 5.96 1.80 3.(50 > 100 > 100 > 100
MALME-3M N.T N.T N.T, N.T. N.T. N.T. 1.98 3.96 7.93 2.15 4.33 - >100 >100 >100
M14 > 100 > 100 > 100 1.85 3.44 6.40
1.83 3.35 6.14 1.83 3.53 6.82 > 100 > 100 > 100
MDA-MB-435 > 100 > 100 > 100 1.71 3.22 6.05
1.82 3.59 1.59 2.99 5.62 > 100 > 100 > 100
SR-MEL-2 16.3 43.2 > 100 1.80 3.32 6.12 198
3.95 7.87 2.05 3.77 6.90 10.60 31.90 96.50
SK-MEL-28 > 100 > 100 > 100 1,81 3.27 5.89
1.75 3.15 5.66 1.89 3.56 > 100 > 100
SK-MEL-5 > 100 > 100 1.57 3.59 8.21 1.51
2.87 5.44 1.60 3.00 5.63 4.23 > 100 > 100
UACC-257 > 100 > 100 > 100 1.74 3.21 5.93
1.90 3.56 6.70 1.86 3.44 6.33 > 100 > 100 > 100
UACC-62 27' >100>,100 1.70.. 3 16 ., .. 5 88 .
.1..67 . . 1,30 .. ..,6,52 .. .11.1 . .. 3 40 _ 6 39 16 90,
. >400 >.1,00 ,
NOWSINCit'' ,..-, -J7'' V '-r, '''' ',:'1==,..,i - , - ,- WO- ....
).:,-. ..:. 1 '.. ...i=!'i= '.':= -.g 1-.10,1111111:-SiY = '.. .. .-
,:"4...... ,.. :..,.:!'1:;:?;.:.'...i.:-.1,B
IGROV1 11.90 54,40 > , 00 1.70 3.34 6.58
161 3.45 1.66 3.. 4.64 > 100 > 100
OVCAR-3 5240 >100 >100 1.61 3.00 5.58 N.T.
N.T. N.T. 1.79 3.51 - N.T. N.T. N.T.
OVCAR-4 > 100 > 100 > 100 1.19 235 6.35
1.52 3.15 6.54 1.60 3.23 15.00 3.74
OVCAR-5 > 100 > 100 > 100 1.45 2.91 5.83
1.46 2.88 5.66 1.63 3.50 7.53 > 100 > 100 > 100
OVCAR-8 17.70 > 100 > 100 1.64 3.15
6.06 1.89 4.12 1.75 3.32 7.75 > 100 > 100
NCl/ADR-RES 33.50 > 100 > 100 1.67 3.17 6.00
1.88 3.67 - 1.82 3.63 . 43.00 > 100 > 100
SK-OV-3 . 11.10 35, 20 _., > 1nn 1 6 3.065 58 , ..
1 '8_ ,3.38 .. 641 1 68 . 3.38 6.80 3.56 _..,... 13 90 ..
..100
1.4-i;Z,. '.'- .: .-' r - ', -' ''V ' 4-1'..
''',.' If.l'.:' -,' ' ,i,, ..i 9;tit 'T ''t. 1
7.,.r...:',,,S;E:".li&
786-0 10.6 33.8 > 100 1.84 3.45 6.46
1.76 3.20 5.81 1.72 3,0 9.55 .18.70 > 100 '
A498 15.3 91.3 > 100 2.70 9.44 3.52 145
4.99 29.20 1.27 3.62 15.00 8.05 3920 > 100
ACHN 8.66 > 100 > 100 1.63 3.07 5.77
1.44 236 5.29 1.67 3.22 4.02 > 100 > 100
CAKI-1 26.4 > 100 > 100 1.43 2.80 5.49
1.56 3.27 1.33 2.69 5.45 6.07 > 100 > 100
RXF 393 13.1 44.6 > 100 1.60 3.41 7.29
N.T. N.T. N.T. 1.64 3.20 N.T. NT. N.T.
SN12C 75.3 > 100 > 100 1.59 3.12 6.12
1.45 302 1.70 3.39 6.80 24.90 > 100 > 100
TK-10 NT N.T N.T. N.T. N.T. N.T. 206
3.58 6.21 2.09 3.74 6.69 5.02 22.30 > 100
L30-31 29.4 > 100 > 100 1 19 2 50 5,22 1
29 2 78 5.96 1.36 2.94 A 35 57.80 > 100 > 100
WHIMBINEIMIBMWErr'.#111 '= = - 'ffilli'...--:''Ir 'jj .! = :-'- - ''- 7' '' =
S If .1fellif!i'.7311:1., ,,- : 77,7i1:::
PC-3 >100 >100 I 1 17 200 5 -4 I '---
29> 570 1.51 1' .: - '...i 5680 >100 > 100
DU-145 > 100 > 100 > 100 1 87 3.34 5.99
1.81 3.20. 5.66 1 76 , , l 6.25 5.50 > 100 > 100
MINIMISM11101111V. t.
'' " ',1',,111S1''''.11-.!11"'., 7-1 '''7".-7 17 F...;' 1;'"r.,7. .7F,. i. ' -
'-7 ' .:'.' - '-, --' !'.' :,,oti, AIN.
..... :
MCF7 0.047 >100 >100 1.23 2.66 5.71 1.28
2.81 1.05 ;...00 a04 . 18.60 > 100
MDA-MB-231/ATCC 10.10 65.5 > 100 1.20 2.51 5.28
1.31 2.65 5.35 1.36 2.86 6.02 236 34.80 > 100
HS 578T 9.64 65,5 > 100 1.74 4.02 9.27 1.69
3.90 1.29 3.53 9.64 2.94 21.20 > 100
BT-549 9.87 > 100 > 100 8.91 2.27 5.33
2.15 5.35 62.00 1.67 3.15 ay 50 > 100 > 100
T-47D NT N.T. N.T. N.T. N.T N.T. 1.56
3.37 1.50 3.43 - 1.26 7.11 > 100
MDA-MB-468 0.040 > 100 > 100 1.47 3.28 7.34 127
2 80 6.15 1 06 2.56 6 20 0.03 0.66 63.10
_
ilifteadannittgiirlitt''
. '-': 4,31 9.ii:i.-AL:'4:1:10;k4.t .04'k1tfc=ii:,-,AN., .1!:..114,!:4,i 1.*
h:.4):tUilik t 'i14,211 ' 53:76
63
