CA2866502C - Thiochromeno(2,3-c)quinolin-12-one derivatives, preparation method and application thereof - Google Patents

Abstract

The invention provides a series of novel thiochromeno[2,3-c] quinolin-12-one derivatives. Further, the invention also provides the preparation method and application of said derivatives, said application comprises: said derivatives with treating effective amount are prepared into pharmaceutical compositions for inhibition of topoisomerase type I and II, inhibition of cancer cell growth, further treating cancer.

Description

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, 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 , , 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 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):

CI =s N
(I) wherein the R is selected from the groups consisting of:
i) halo, amino, hydroxyl and thiol groups;

4 . , 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 . .
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 (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 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 . , . , -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 . .
. .
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 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 , .
. .
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 . .
. .
-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;

(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 , 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, ) 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.

=
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 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 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 (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 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 . .
. , (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).

. =
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.

, 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 [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, , .
. .
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, 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 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.

. , . .
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 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, . , . .
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) . , . .
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) 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 , .
. .
-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) . .
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, 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.

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.

. .
= 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, = 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 , 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 = .
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=

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) 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=

. .
= .
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 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, =
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 =

=
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]+:

. .
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, . , 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 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, 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 , , 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) 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 . .
. .
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 > 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 \

, .
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 / /
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

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

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 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 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

. .
. .
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 AN 0 > 11.82 5.83 >15 AN
N28 H 1.1 9.61 4.88 6.01 3.31 N29 AN 0 C) 4.86 0.99 6.20 2.52 N30 AN 10 >15 >15 H

N31 i >15 >15 TN
H

N32 ,4 13.59 1.39 10.11 0.94 H

N33i ,..1,, 8.11 1.37 6.13 6.98 r- N NH2 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 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 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.

. .
. .
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' 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.

.., .
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' :2 Panel/Cell line 4, -r- F-- 'Fn-in cn z in in Leukemia ______________________ ..
CCRF-CEIS4 79.77 88.62 93.72 -53.54 81.07 70.00 98.57 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.

. .
. .
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*

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.

. .
. .
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.

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

Claims (12)

What is claimed is:

1. A compound as shown in formula (I):
wherein R is:
i) a halo, amino, hydroxyl or thiol group;
ii) a linear alkyl chain of NH(CH2)nH, an alkyl group with substituted side chains, an alkyl side chain with a substituted amino group or an alkyl side chain with a substituted hydroxyl group, wherein 1 <= n <=
10;
iii) O(CH2)nH, N(CH3)2, or NH(CH2)nNH(CH2)nOH, wherein 1 <= n <= 10;
iv) a nitrogen-containing cycloalkyl group or an heterocyclic compound of C3-12 which contain 1 to 3 heteroatoms which are O, S or N, wherein at the ortho-, para- and meta- position is independently an hydrogen group, (CH2)n alkyl group, (CH2)n hydroxyl group, (CH2)n3-12 cycloalkyl group, (CH2)nC3-12 nitrogen-containing cycloalkyl group, (CH2)n benzene ring, formyl group or (CH2)nCOC3-12 nitrogen-containing cycloalkyl group, wherein 0 <= n <= 10;
v) NH(CH2)nR1, 0 <= n <=, wherein R1 is N(CH3)2, C(NH2)2, linear alkyl chain of NH(CH2)nH, alkyl group with substituted side chains, alkyl side chains with a substituted amino group or alkyl side chains with a substituted hydroxyl group;
vi) NH(CH2)nR2, 0 <= n <= 10, wherein R2 is a benzene ring, cycloalkyl group or heterocyclic group of which contain 1 to 3 heteroatoms which are O, S or N, wherein at the ortho-, para- and meta- position is independently a methoxyl group, amino group, benzene ring, alkyl, amino, nitro,hydroxyl group with substituted C1-C3 side chains or C3-12 heterocyclic group; wherein the C3-12 heterocyclic group contains 1 to 3 heteroatoms which are O, S or N;
and their pharmaceutically acceptable salts, stereoisomers and enantiomers.

2. The compound according to claim 1, wherein the R group of i)-iv) is chlorine, hydroxyl, methoxyl, dimethylamino, piperazin-1-yl, 4-methylpiperazin-1-yl, 4-ethylpiperazin-1-yl, 4-(2-hydroxyethyl)piperazin-1-yl, 4-Benzylpiperazin-1-yl, 4-phenylpiperazin-1-yl, morpholino, thiomorpholino, piperidin-1-yl, 4-hydroxypiperidin-1-yl, 4-Benzylpiperidin-1-yl, (1,4'-Bipiperidin)-1'-yl, 4-(3-(piperidin-4-yl)propyl)piperidin-1-yl, pyrrolidin-1-yl, 2-oxopiperidin-1-yl, 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-Aminoethyl)amino, (2-((2-hydroxyethyl)amino)ethyl)amino, (2-morpholinoethyl)amino, (3-(dimethylamino)propyl)amino, (3-(diethylamino)propyl)amino, (3-((2-hydroxyethyl)amino)propyl)amino, (2,3-dihydro-1H-inden-2-yl)amino, cyclohexylamino, (1-Benzylpiperidin-4-yl)amino, (thiophen-2-ylmethyl)amino, (cyclohexylmethyl)amino, benzylamino, (pyridin-2-ylmethyl)amino, (Benzo[d][1,3]dioxol-5-ylmethyl)amino, (2-methoxybenzyl)amino, (3,4-dimethoxybenzyl)amino, phenethylamino, (4-methoxyphenethyl)amino, (4-aminophenethyl)amino, guanidine, or piperidin-1-ylamino.

3. The compound according to claim 1, wherein the compound is:
3-((4-Chlorophenyl)thio)-2-hydroxyquinoline-4-carboxylic acid, 6,9-Dichloro-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-hydroxy-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-methoxy-12H-thiochromeno[2,3-c]quinolin-12-one10-Chloro-6-dimethylamino-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-(piperazin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-(4-methylpiperazin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-(4-ethylpiperazin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-(4-(2-hydroxyethyl)piperazin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12one, 6-(4-Benzylpiperazin-1-yl)-10-chloro-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-(4-phenylpiperazin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-morpholino-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-thiomorpholino-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-(piperidin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-(4-hydroxypiperidin-1-yl)-12H-thiochromeno[2,3 -c]quinolin-12-one, 6-(4-Benzylpiperidin-1-yl)-10-chloro-12H-thiochromeno[2,3-c]quinolin-12-one, 6-([1,4'-Bipiperidin]- 1'-yl)-10-chloro-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-(4-(3-(piperidin-4-yl)propyl)piperidin-1-yl)-12H-thiochromeno [2,3 -c]quinolin-12-one, 10-Chloro-6-(pyrrolidin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-(2-oxopiperidin-1-yl)-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-ylamino)-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-((2-(dimethylamino)ethyl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-((2-(diethylamino)ethyl)amino)-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-one, 10-Chloro-6-((1-hydroxybutan-2-yl(amino)-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-((4-methylpentan-2yl(amino)-12H-thiochromeno[2,3-c]quinolin-12-one, 6-((2-Aminoethyl)amino)-10-chloro-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-((2-((2-hydroxyethyl)amino)ethyl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-((2-morpholinoethyl(amino)-12H-thiochromeno[2,3-c]quinolin-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-6-((3-((2-hydroxyethyl)amino)propyl)amino)-12H-thiochromeno[2,3-c]quinolin-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-ylmethyl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-((cyclohexylmethyl)amino)-12H-thiochromeno[2,3-c]quinolin-12-, one, 6-(Benzylamino)-10-chloro-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-((pyridin-2-ylmethyl(amino)-12H-thiochromeno[2,3-c]quinolin-12-one, 6-((Benzo[d][1,3]dioxol-5-ylmethyl)amino)-10-chloro-12H-thiochromeno[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-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-(phenethylamino)-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-((4-methoxyphenethyl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one, 6-((4-Aminophenethyl)amino)-10-chloro-12H-thiochromeno[2,3-c]quinolin-12-one, 2-(10-Chloro-12-oxo-12H-thiochromeno[2,3-c]quinolin-6-yl)guanidine, 10-Chloro-6-(piperidin-1-ylamino)-12H-thiochromeno[2,3-c]quinolin-12-one, or their salts.

4. A pharmaceutical composition comprising a compound according to claim 1 and at least one pharmaceutically acceptable vehicle, diluent or excipient.

5. Use of the compound according to claim 1 for inhibiting Topoisomerase I
activity.

6. Use of the compound according to claim 1 in the manufacture of a medicament for inhibiting Topoisomerase I activity.

7. Use of the compound according to claim 1 for inhibiting Topoisomerase II

activity.

8. Use of the compound according to claim 1 in the manufacture of a medicament for inhibiting Topoisomerase II activity.

9. Use of the compound according to claim 1 for treating cancer.

10. Use of the compound according to claim 1 in the manufacture of a medicament for treating cancer.

11. The use according to claim 9 or 10, wherein the cancer is leukemia, non-small cell lung cancer, colorectal cancer, central nervous system (CNS) cancer, melanoma, ovarian cancer, renal cancer, prostate cancer or breast cancer.

12. A method for preparing thiochromeno[2,3-c]quinolin-12-one derivatives, wherein the method comprises:
heating a mixture of isatin, 2-((4-chlorophenyl)thio)acetic acid and sodium acetate at 150 °C for 1 h, cooling the mixture and adding acetic acid thereto, collecting the precipitate, washing the precipitate with acetic acid, water and n-hexane, and obtaining compound 2 (3-((4-Chlorophenyl)thio)-2-hydroxyquinoline-4-carboxylic acid);
heating a solution of compound 2 (3-((4-Chlorophenyl)thio)- 2-hydroxyquinoline-4-carboxylic acid) in phosphoryl trichloride at 150 °C
for 48 h, cooling the mixture and pouring said mixture into water at 0 °C, collecting the precipitate by filtration, adding 10% NaHCO3 thereto with vigorous stirring for 1 h, collecting the resulting precipitate and washing said precipitate with H2O, recrystallizing the crude solid with dichloromethane to obtain compound 3 (6,9-Dichloro-12H-thiochromeno[2,3-c] quinolin-12-one);
adding concentrated HCI to a solution of compound 3 (6,9-Dichloro-12H-thiochromeno [2,3-c]quinolin-12-one) in DMF and refluxing; after 6 hours, adding concentrated HCI dropwise and refluxing for another 12 hours, evaporating the mixture in vacuo and treating said mixture with H2O, filtering the crude solid and washing said crude solid with EtOH to obtain compound 4 (10-Chloro-6-hydroxy-12H-thiochromeno[2,3-c]quinolin-12-one);
refluxing a suspension of compound 3 (6,9-Dichloro-12H-thiochromeno[2,3-c]quinolin-12-one) and sodium methoxide in methanol for 16 h, cooling and removing the solvent, filtrating and washing with ethanol and n-hexane to collect compound 5 (10-Chloro-6-methoxy-12H- thiochromeno[2,3-c]quinolin-12-one);
refluxing a solution of compound 3 (6,9-Dichloro-12H- thiochromeno[2,3-c]quinolin-12-one), suitable secondary amines and sodium carbonate in DMSO
for 10 hours, adding ice water, filtering the precipitate, washing with water/methanol and collecting the following:

10-Chloro-6-dimethylamino-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-(piperazin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-(4-methylpiperazin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-(4-ethylpiperazin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-(4-(2-hydroxyethyl)piperazin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12one, 6-(4-Benzylpiperazin-1-yl)-10-chloro-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-(4-phenylpiperazin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-morpholino-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-thiomorpholino-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-(piperidin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-(4-hydroxypiperidin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one, 6-(4-Benzylpiperidin-1-yl)-10-chloro-12H-thiochromeno[2,3-c]quinolin-12-one, 6-([1,4'-Bipiperidin]-1'-yl)-10-chloro-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-(4-(3-(piperidin-4-yl)propyl)piperidin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-(pyrrolidin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one, or 10-Chloro-6-(2-oxopiperidin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one respectively;

refluxing a solution of compound 3 (6,9-Dichloro-12H- thiochromeno[2,3-c]quinolin-12-one) in DMSO and suitable primary amines for 8 hours, cooling the solution and then adding water, filtering the precipitate and washing with water and methanol to collect the following:
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-ylamino)-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-((2-(dimethylamino)ethyl(amino)-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-((2-(diethylamino)ethyl(amino)-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-one, 10-Chloro-6-((1-hydroxybutan-2-yl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-((4-methylpentan-2-yl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one, 6-((2-Aminoethyl)amino)-10-chloro-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-((2-((2-hydroxyethyl)amino)ethyl)amino)-12H-thiochromeno [2,3 -c] quinolin-12-one, 10-Chloro-6-((2-morpholinoethyl(amino)-12H-thiochromeno[2,3-c] quinolin-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-6-((3 -((2-hydroxyethyl)amino)propyl)amino)-12H-thiochromeno[2,3 -c]quinolin-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-ylmethyl)amino)- 12H-thiochromeno[2,3-c] quinolin-12-one, 10-Chloro-6-((cyclohexylmethyl(amino)-12H-thiochromeno[2,3-c] quinolin-12-one, 6-(Benzylamino)-10-chloro-12H-thiochromeno [2,3-c] quinolin-12-one, 10-Chloro-6-((pyridin-2-ylmethyl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one, 6-((Benzo[d][1,3]dioxo1-5-ylmethyl)amino)-10-chloro-12H-thiochromeno[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-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-(phenethylamino)-12H-thiochromeno[2,3-c]quinolin-12-one, 10-Chloro-6-((4-methoxyphenethyl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one, 6-((4-Aminophenethyl)amino)-10-chloro-12H-thiochromeno[2,3-c]quinolin-12-one, 2-(10-Chloro-12-oxo-12H-thiochromeno[2,3-c]quinolin-6-yl)guanidine, or 10-Chloro-6-(piperidin-1-ylamino)-12H-thiochromeno[2,3-c]quinolin-12-one respectively.