PATENT DOCUMENT

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.

Full Description:
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 (M1 stage). When a tumor suppressor gene such as p53 or Rb is mutated within M1 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 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 stage. 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 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-c]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): 
                                
wherein the R is selected from the groups consisting of:
         i) haloformyl, amino, hydroxyl and thiol groups;   ii) linear alkyl chains of N(CH 2 ) n H, 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) O(CH 2 ) n H, N(CH 3 ) 2 , NH(CH 2 ) n NH(CH 2 ) n OH, wherein 1≦n≦10;   iv) nitrogen-containing cycloalkyl groups and heterocyclic compounds of C 3-12  which contain 1 to 3 heteroatoms selected from O, S and N, wherein the ortho-, para- and meta-position can be further selected independently from one of the groups consisting of: hydrogen group, (CH 2 ) n  alkyl groups, (CH 2 ) n  hydroxyl groups, (CH 2 ) n C 3-12  cycloalkyl groups, (CH 2 ) n C 3-12  nitrogen-containing cycloalkyl groups, (CH 2 ) n  benzene rings, formyl group and (CH 2 ) n COC 3-12  nitrogen-containing cycloalkyl groups, wherein 0≦n≦10;   v) NH(CH 2 ) n R 1 , 0≦n≦10, wherein R 1  is selected from the groups consisting of: N(CH 3 ) 2 , C(NH 2 ) 2 , linear alkyl chains of NH(CH 2 ) n H, 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(CH 2 ) n R 2 , 0≦n≦10, wherein R 2  is selected from the groups consisting of: benzene rings, C 3-12  cycloalkyl groups and heterocyclic groups of which contain 1 to 3 heteroatoms selected from O, 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 substituted C1-C3 side chains and C 3-12  heterocyclic groups; wherein the C 3-12  heterocyclic groups which contain 1 to 3 heteroatoms selected from O, S and N;   and their pharmaceutically acceptable salts, stereoisomers and enantimoers.       

     According to the invention, wherein the functional group i)˜iv) are selected from the group consisting of 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 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-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-one   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-ylpropyl)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-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]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,
 
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 effective 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 invention, wherein the cancers are 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% NaHCO 3  with vigorous stirring for 1 h, the resulting precipitate was collected and washed with H 2 O, 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. HCl and refluxed, after 6 hours, the conc. HCl was added dropwise and refluxed for another 12 hours, the mixture was evaporated in vacuo and treated with H 2 O, after filtered the crude solid was washed with EtOH 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]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, and   10-Chloro-6-(2-oxopiperidin-1-yl)-12H-thiochromeno[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-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]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, and   10-Chloro-6-(piperidin-1-ylamino)-12H-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]quinolin-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 μM and 50 μM. 
         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 μM. 
         FIG. 4   a - 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 μM and 50 μM. 
         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 50 μM. 
         FIG. 7   a - 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&#39;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 F 254 . Melting points of all synthetic compounds were measured with Büchi B-545 melting point apparatus.  1 H NMR: Varian GEMINI-300 (300 MHz) or Agilent 400 MR DD2 (400 MHz); δ 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% NaHCO 3  solution (50 mL) with vigorous stirring for 1 h. The resulting precipitate was collected and washed with H 2 O. 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. HCl (3 mL) and refluxed. After 6 h, the conc. HCl (6 mL) was added dropwise and refluxed for another 12 h. The mixture was evaporated in vacuo and treated with H 2 O (20 mL), after filtered the crude solid was washed with EtOH 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 (0.55 g, 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-Chlorophenyl)thio)-2-hydroxyquinoline-4-carboxylic acid (TC-SCl) (2) 
     The pure compound was obtained as a gray solid (yield 86%). (R f =0.5 at EA:AcOH=20:1). Mp 306-308° C.  1 H NMR (300 MHz, DMSO-d 6 ): δ (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, Ar—H), 7.62 (1H, t, J=8.0 Hz, Ar—H), 12.22 (1H, s, —COOH).  13 C NMR (100 MHz, DMSO-d 6 ): δ (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 ( C O). HRMS (ESI) calcd for C 16 H 10 NO 3 SCl [M] +  331.0070. found [M+H] + 332.0147 (100), [M+H+2] +  334.0122 (33). found [M−H] −  330.0002. 
     Example 2 
     6,9-Dichloro-12H-thiochromeno[2,3-c]quinolin-12-one (3) 
     The yellow solid material was isolated in 90% yield (R f =0.50 at CH 2 Cl 2 :n-hexane=1:1). Mp 259-261° C. (CH 2 Cl 2 ).  1 H NMR (400 MHz, CDCl 3 ): δ (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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) calcd for C 16 H 7 NOSCl 2  [M] +  330.9625. found [M+H] +  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-c]quinolin-12-one (4) 
     The yellow solid material was isolated in 95% yield (R f =0.40 at EA). Mp&gt;410° C.  1 H NMR (400 MHz, DMSO-d6): δ 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).  13 C NMR (75 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) m/z calcd for C 16 H 8 NO 2 SCl [M] + : 312.9964. found, 314.0051. 
     Example 4 
     10-Chloro-6-methoxy-12H-thiochromeno[2,3-c]quinolin-12-one (5) 
     The gray solid material was isolated in 91% yield (R f =0.52 at CH 2 Cl 2 :n-hexane=1:1). Mp 227-228° C.  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 4.27 (3H, s, —OCH 3 ), 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).  13 C NMR (100 MHz, CDCl 3 ): δ (ppm) 54.83 (OCH 3 ), 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 ( C O). HRMS (ESI) m/z calcd for C 17 H 10 NO 2 SCl [M] +  327.0121. found [M+H] +  328.0203, [M+H+2] +  330.0172. 
     Example 5 
     10-Chloro-6-dimethylamino-12H-thiochromeno[2,3-c]quinolin-12-one (6) 
     Product 6 was prepared from 3 and dimethylamine. The light-yellow solid material was isolated in 85% yield (R f =0.45 at CH 2 Cl 2 :n-hexane=1:1). Mp 194-195° C.  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 3.06 (6H, s, —CH 3 ), 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) calcd for C 18 H 13 N 2 OSCl [M] +  340.0437. found [M+H] +  341.0517 (100), [M+H+2] +  343.0501 (33). 
     Example 6 
     10-Chloro-6-(piperazin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one (7) 
     Product 7 was prepared from 3 and piperazine. The dark-yellow solid material was isolated in 69% yield (R f =0.12 at EA:MeOH:ammonia water=20:5:1). Mp 211-213° C.  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 3.20 (4H, t, J=4.8 Hz, —CH 2 —), 3.36 (4H, t, J=4.8 Hz, —CH 2 —), 7.60-7.66 (3H, 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) calcd for C 20 H 16 N 3 OSCl [M] + 381.0703. found [M+H] +  382.0783. 
     Example 7 
     10-Chloro-6-(4-methylpiperazin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one (8) 
     Product 8 was prepared from 3 and 1-methylpiperazine. The green-yellow solid material was isolated in 80% yield (R f =0.24 at EA:methanol=5:1). Mp 212-214° C.  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 2.51 (3H, s, —CH 3 ), 2.84 (4H, br, —CH 2 —), 3.50 (4H, t, J=4.5 Hz, —CH 2 —), 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) calcd for C 21 H 18 N 3 OSCl [M] + 395.0859. found [M+H] + 396.0926. 
     Example 8 
     10-Chloro-6-(4-ethylpiperazin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one (9) 
     Product 9 was prepared from 3 and 1-ethylpiperazine. The yellow solid material was isolated in 74% yield (R f =0.48 at EA:MeOH=10:1). Mp 196-198° C.  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 1.19 (3H, t, J=7.2 Hz, —CH 3 ), 2.58 (2H, q, J=7.2 Hz, —CH 2 —), 2.78 (4H, br, —CH 2 —), 3.46 (4H, t, J=4.4 Hz, —CH 2 —), 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) calcd for C 22 H 20 N 3 OSCl [M] + 409.1016. found [M+H] +  410.1069. 
     Example 9 
     10-Chloro-6-(4-(2-hydroxyethyl)piperazin-1-yl)-12H-thiochromeno[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 (R f =0.37 at EA:MeOH=2:1). Mp 211-213° C.  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 2.74 (2H, t, J=5.2 Hz, —CH 2 —), 2.87 (4H, t, J=3.6 Hz, —CH 2 —), 3.45 (4H, t, J=3.6 Hz, —CH 2 —), 3.72 (2H, t, J=5.2 Hz, —CH 2 O—), 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).  13 C NMR (100 MHz, CDCl 3 ): δ (ppm) 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 ( C O). HRMS (ESI) calcd for C 22 H 20 N 3 O 2 SCl [M] +  425.0965. found [M+H] +  426.1024. 
     Example 10 
     6-(4-Benzylpiperazin-1-yl)-10-chloro-12H-thiochromeno[2,3-c]quinolin-12-one (11) 
     Product 11 was prepared from 3 and 4-benzylpiperazine. The yellow solid material was isolated in 81% yield (R f =0.43 at EA:n-hexane=1:4). Mp 191-193° C.  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 2.78 (4H, br, —CH 2 N—), 3.43 (4H, t, J=4.85 Hz, —NCH 2 —), 3.68 (2H, s, —CH 2 —), 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) calcd for C 27 H 22 N 3 OSCl [M] +  471.1172. found [M+H] +  472.1241. 
     Example 11 
     10-Chloro-6-(4-phenylpiperazin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one (12) 
     Product 12 was prepared from 3 and 1-phenylpiperazine. The yellow solid material was isolated in 77% yield (R f =0.73 at EA:n-hexane=1:4). Mp 236-237° C.  1 H NMR (300 MHz, CDCl 3 ): δ (ppm) 3.50-3.60 (8H, m, —CH 2 —), 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).  13 C NMR (75 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) calcd for C 26 H 20 N 3 OSCl [M] +  457.1016. found [M+H] +  458.1095. 
     Example 12 
     10-Chloro-6-morpholino-12H-thiochromeno[2,3-c]quinolin-12-one (13) 
     Product 13 was prepared from 3 and morpholine. The yellow solid material was isolated in 70% yield (R f =0.42 at CH 2 Cl 2 ). Mp 217-218° C.  1 H NMR (300 MHz, CDCl 3 ): δ (ppm) 3.41 (4H, t, J=4.5 Hz, —NCH 2 —), 4.02 (4H, t, J=4.5 Hz, —CH 2 O—), 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).  13 C NMR (75 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) calcd for C 20 H 15 N 2 O 2 SCl [M] +  382.8633. found [M+H] +  383.0620. 
     Example 13 
     10-Chloro-6-thiomorpholino-12H-thiochromeno[2,3-c]quinolin-12-one (14) 
     Product 14 was prepared from 3 and thiomorpholine. The yellow solid material was isolated in 86% yield (R f =0.77 at EA:n-hexane=1:4). Mp 219-220° C.  1 H NMR (300 MHz, CDCl 3 ): δ (ppm) 2.98 (4H, t, J=4.8 Hz, —NCH 2 —), 3.64 (4H, t, J=5.1 Hz, —SCH 2 —), 7.62-7.84 (3H, m, Ar—H), 7.73 (1H, 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).  13 C NMR (75 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) m/z calcd for C 20 H 15 N 2 S 2 OCl + [M] +  398.0314. found [M+H] +  399.0420, [M+H+2] +  401.0394. 
     Example 14 
     10-Chloro-6-(piperidin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one (15) 
     Product 15 was prepared from 3 and piperidine. The yellow solid material was isolated in 86% yield (R f =0.75 at EA). Mp 187-188° C.  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 1.71-1.74 (2H, m, —CH 2 —), 1.88 (4H, p, J=4.5 Hz, —CH 2 —), 3.31 (4H, t, J=4.2 Hz, —NCH 2 —), 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=1.5 Hz, Ar—H), 9.62 (1H, dd, J=6.3, 0.6 Hz, Ar—H)  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) calcd for C 21 H 17 N 2 OSCl [M] +  380.0750. found [M+H] +  381.0816. 
     Example 15 
     10-Chloro-6-(4-hydroxypiperidin-1-yl)-12H-thiochromeno[2,3-c ]quinolin-12-one (16) 
     Product 16 was prepared from 3 and 4-hydroxypiperidine. The gray-yellow solid material was isolated in 84% yield (R f =0.4 at EA:n-hexane=1:1). Mp 224-225° C.  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 1.89 (1H, td, J=7.2, 2.7 Hz, piperidine-CH a ), 1.94 (1H, td, J=6.9, 2.7 Hz, piperidine-CH a ), 2.15-2.21 (2H, m, piperidine-CH e ), 3.19 (2H, td, J=8.4, 2.1 Hz, piperidine-NCH a ), 3.60-3.65 (2H, m, piperidine-NCH e ), 4.01 (1H, sext, J=3.0 Hz, piperidine-CH), 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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, 144.91, 157.83, 181.47 ( C O). HRMS (ESI) calcd for C 21 H 17 N 2 OSCl [M] +  396.0699. found [M+H] +  397.0757. 
     Example 16 
     6-(4-Benzylpiperidin-1-yl)-10-chloro-12H-thiochromeno[2,3-c]quinolin-12-one (17) 
     Product 17 was prepared from 3 and 4-benzylpiperidine. The yellow solid material was isolated in 90% yield (R f =0.57 at CH 2 Cl 2 :n-hexane=2:1). Mp 184-185° C.  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 1.67 (2H, td, J=9.3, 3.0 Hz, —CH 2 —), 1.79-1.89 (1H, m, —CH—), 1.88 (2H, d, J=6.9 Hz, piperidine-CH 2 ), 2.71 (2H, d, J=5.1 Hz, piperidine-CH 2 ), 3.00 (2H, td, J=9.3, 1.2 Hz, —NCH 2 —), 3.65 (2H, d, J=9.3 Hz, —NCH 2 —), 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) calcd for C 28 H 23 N 2 OSCl [M] +  471.0130. found [M+H] +  471.1276. 
     Example 17 
     6-([1,4′-Bipiperidin]-1′-yl)-10-chloro-12H-thiochromeno[2,3-c]quinolin-12-one (18) 
     Product 18 was prepared from 3 and 1,4′-bipiperidine. The yellow solid material was isolated in 92% yield (R f =0.15 at EA:MeOH=5:1). Mp 187-189° C.  1 H NMR (400 MHz, CDCl 3 ): δ (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).  13 C NMR (100 MHz, CDCl 3 ): δ (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, 133.56, 134.54, 144.92, 157.92, 181.47 ( C O). HRMS (ESI) calcd for C 26 H 26 N 3 OSCl [M] +  463.1485. found [M+H] +  464.1593. 
     Example 18 
     10-Chloro-6-(4-(3-(piperidin-4-yl)propyl)piperidin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one (19) 
     Product 19 was prepared from 3 and 1,3-di(piperidin-4-yl)propane. The yellow solid material was isolated in 76% yield (R f =0.13 at CH 2 Cl 2 ). Mp 164-165° C.  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 1.16 (2H, qd, J=11.6, 3.2 Hz, —CH 2 —), 1.20-1.28 (2H, m, —CH 2 —), 1.36-1.39 (4H, m, —CH 2 —), 1.51-1.58 (4H, m, —CH 2 —), 1.70 (2H, d, J=13.6 Hz, —CH 2 —), 1.87 (2H, d, J=9.6 Hz, —CH2-), 2.43 (1H, br, —NH), 2.60 (2H, td, J=12.0, 2.0 Hz, —CH 2 —), 2.99 (2H, t, J=11.2 Hz, —CH 2 —), 3.10 (2H, d, J=12 Hz, —CH 2 —), 3.64 (2H, d, J=12.4 Hz, —CH 2 —), 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, Ar—H).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) calcd for C 29 H 32 N 3 OSCl [M] +  505.1955. found [M+H] +  506.2004. 
     Example 19 
     10-Chloro-6-(pyrrolidin-1-yl)-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 (R f =0.56 at CH 2 Cl 2 :n-hexane=1:1). Mp 170-171° C.  1 H NMR (300 MHz, CDCl 3 ): δ (ppm) 2.05 (4H, quin, J=3.6 Hz, —CH 2 —), 3.76 (4H, t, J=6.9 Hz, —NCH 2 —), 7.50 (1H, td, J=7.2, 1.5 Hz, Ar—H), 7.61 (1H, d, J=1.5 Hz, Ar—H 11 ), 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).  13 C NMR (75 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) calcd for C 20 H 15 N 2 OSCl [M] +  366.0594. found [M+H] +  367.0659. 
     Example 20 
     10-chloro-6-(2-oxopiperidin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one (TC-SCl-B-18) (21) 
     Product 21 was a yellow solid material which was isolated in 89% yield. Mp: 258-261° C.  1 H NMR (400 MHz, CDCl 3 ): δ ppm. 1.25 (1H, d, J=4.8 Hz, piperidone-H), 2.44 (2H, quin, —CH 2 —), 2.61 (1H, s, piperidone-H), 2.75 (2H, t, J=8.4 Hz, —CH 2 —), 4.11-4.14 (2H, m, —CH 2 —), 7.60-7.67 (2H, m, Ar—H), 7.78-7.81 (2H, m, Ar—H), 8.09-8.12 (1H, m, Ar—H), 8.60 (1H, d, J=2.0 Hz, Ar—H), 9.72-9.75 (1H, m, Ar—H).  13 C NMR (100 MHz, CDCl 3 ): δ 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. 
     Example 21 
     10-Chloro-6-methylamino-12H-thiochromeno[2,3-c]quinolin-12-one (N1) 
     Product N1 was prepared from 3 and methylamine. The pure compound was obtained as a yellow solid (yield 92%) (R f =0.65 at CH 2 Cl 2 ). Mp 237-238° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 3.26 (3H, d, J=4.8 Hz, —CH 3 ), 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) m/z calcd for C 17 H 11 N 2 OSCl [M] + : 326.0281. found [M+H] + : 327.0356. 
     Example 22 
     10-Chloro-6-ethylamino-12H-thiochromeno[2,3-c]quinolin-12-one (N2) 
     Product N2 was prepared from 3 and ethylamine. The pure compound was obtained as a yellow solid (yield 91%) (R f =0.75 at CH 2 Cl 2 ). Mp 204-205° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 1.41 (3H, t, J=7.2 Hz, —CH 3 ), 3.75 (2H, q, J=1.6 Hz, —CH 2 ), 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) m/z calcd for C 18 H 13 N 2 OSCl [M] + : 340.0437. found [M+H] + : 341.0493. 
     Example 23 
     10-Chloro-6-propylamino-12H-thiochromeno[2,3-c]quinolin-12-one (N3) 
     Product N3 was prepared from 3 and propylamine. The pure compound was obtained as a yellow solid (yield 85%) (R f =0.82 at CH 2 Cl 2 ). Mp 178-179° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 1.09 (3H, t, J=7.2 Hz, —CH 3 ), 1.81 (2H, sext, J=7.2 Hz, —CH 2 —), 3.69 (2H, q, J=7.2 Hz, —NCH 2 —), 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) m/z calcd for C 19 H 15 N 2 OSCl [M] + : 354.0594. found [M+H] + : 355.0651. 
     Example 24 
     6-(Butylamino)-10-chloro-12H-thiochromeno[2,3-c]quinolin-12-one (N4) 
     Product N4 was prepared from 3 and butylamine. The pure compound was obtained as a yellow solid (yield 91%) (R f =0.85 at CH 2 Cl 2 ). Mp 147-149° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 1.03 (3H, t, J=7.2 Hz, —CH 3 ), 1.53 (2H, sext, J=7.2 Hz, —CH 2 —), 1.76 (2H, quin, J=7.2 Hz, —CH 2 —), 3.71 (2H, q, J=6.8 Hz, —NCH 2 —), 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 (1H, d, J=8.4 Hz, Ar—H).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) m/z calcd for C 20 H 17 N 2 OSCl [M] + : 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%) (R f =0.85 at CH 2 Cl 2 ). Mp 159-160° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 1.08 (6H, d, J=6.8 Hz, —CH 3 ), 2.10 (1H, sep, J=6.8 Hz, —CH), 3.56 (2H, t, J=6.4 Hz, —CH 2 —), 4.94 (1H, 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) m/z calcd for C 20 H 17 N 2 OSCl [M] + : 368.0750. found [M+H] + : 369.0825. 
     Example 26 
     10-Chloro-6-(pentan-3-ylamino)-12H-thiochromeno[2,3-c]quinolin-12-one (N6) 
     Product N6 was prepared from 3 and 3-aminopentane. The pure compound was obtained as a light yellow crystal (yield 65%) (R f =0.87 at CH 2 Cl 2 ). Mp 160-161° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 0.92 (6H, t, J=7.6 Hz, —CH 3 ), 1.69 (4H, quin, J=6.0 Hz, —CH 2 —), 4.34 (1H, sext, J=7.2 Hz, —CH—), 6.58 (1H, 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, 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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, 151.64, 180.85 ( C O). HRMS (ESI) m/z calcd for C 21 H 19 N 2 OSCl [M] + : 382.0907. found [M+H] + : 383.0994, [M−H] − : 381.0851. 
     Example 27 
     10-Chloro-6-((2-(dimethylamino)ethyl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one (N7) 
     Product N7 was prepared from 3 and N,N-dimethylethylenediamine. The pure compound was obtained as a yellow crystal (yield 76%) (R f =0.82 at EA:MeOH:ammonia water=10:5:1). Mp 156-157° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 2.36 (6H, s, —N(CH 3 ) 2 ), 2.69 (2H, t, J=6.0 Hz, —CH 2 N—), 3.57 (2H, q, J=5.6 Hz, —NCH 2 —), 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).  13 C NMR (100 MHz, CDCl 3 ): δ (ppm) 39.53, 45.28, 57.57, 120.71, 123.46, 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. ( C O). HRMS (ESI) m/z calcd for C 20 H 18 N 3 OSCl [M] + : 383.0859. found [M+H] + : 384.0925. 
     Example 28 
     10-Chloro-6-((2-(diethylamino)ethyl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one (N8) 
     Product N8 was prepared from 3 and N,N-diethylethylenediamine. The pure compound was obtained as a yellow crystal (yield 86%) (R f =0.8 at EA:MeOH:ammonia water=10:5:1). Mp 152-153° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 1.13 (6H, t, J=7.2 Hz, —CH 3 ), 2.64 (4H, q, J=6.8 Hz, —NCH 2 —), 2.82 (2H, t, J=6.0 Hz, —CH 2 N—), 3.70 (2H, q, J=5.2 Hz, —NCH 2 —), 6.08 (1H, br, —NH—), 7.43 (1H, t, J=7.2 Hz, Ar—H), 7.59-7.64 (3H, 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) m/z calcd for C 22 H 22 N 3 OSCl [M] + : 411.1172. found [M+H] + : 412.1262. 
     Example 29 
     10-Chloro-6-(2-ethanolamino)-12H-thiochromeno[2,3-c]quinolin-12-one (N9) 
     Product N9 was prepared from 3 and ethanolamine. The pure compound was obtained as a yellow crystal (yield 77%) (R f =0.65 at EA). Mp 190-192° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 3.93 (2H, q, J=4.4 Hz, —NCH 2 —), 4.00 (2H, t, J=4.4 Hz, —CH 2 O—), 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) m/z calcd for C 18 H 13 N 2 O 2 SCl [M] + : 356.8260. found [M+H] + : 357.0476, [M+H+2] + : 359.0455. 
     Example 30 
     10-Chloro-6-(3-propanolamino)-12H-thiochromeno[2,3-c]quinolin-12-one (N10) 
     Product N10 was prepared from 3 and 3-amino-1-propanol. The pure compound was obtained as a yellow solid (yield 94%) (R f =0.66 at EA). Mp 201-202° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 1.94 (2H, p, J=6.0 Hz, —CH 2 —), 3.72 (2H, t, J=5.2 Hz, —NCH 2 —), 3.93 (2H, q, J=6.0 Hz, —CH 2 O—), 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), 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) m/z calcd for C 19 H 15 N 2 O 2 SCl [M] + : 370.0543. found [M+H] + : 371.0622. 
     Example 31 
     10-Chloro-6-(5-pentanolamino)-12H-thiochromeno[2,3-c]quinolin-12-one (N11) 
     Product N11 was prepared from 3 and 5-amino-1-pentanol. The pure compound was obtained as a yellow solid (yield 91%) (R f =0.7 at EA). Mp 158-160° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 1.40 (1H, br, —OH), 1.49-1.62 (2H, m, —CH 2 —), 1.71 (2H, quin, —CH 2 —), 1.83 (2H, quin, —CH 2 —), 3.74 (4H, quin, —CH 2 —), 4.91 (1H, br, —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).  13 C NMR (100 MHz, CDCl 3 ): δ (ppm) 23.39, 29.17, 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 ( C O). HRMS (ESI) m/z calcd for C 21 H 19 N 2 O 2 SCl [M] + : 398.0856. found [M+H] + : 399.0914. 
     Example 32 
     10-Chloro-6-((1-hydroxybutan-2-yl)amino)-12H-thiochromeno[2,3-c]quinolin-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%) (R f =0.8 at EA). Mp 203-204° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 1.21 (3H, t, J=7.6 Hz, —CH 3 ), 1.71-1.88 (2H, m, —CH 2 —), 3.79 (1H, dd, J=11.2, 1.6 Hz, —CH 2 —), 3.99 (1H, dd, J=11.2, 2.8 Hz, —CH 2 —), 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) m/z calcd for C 20 H 17 N 2 O 2 SCl [M] + : 384.0699. found [M+H] + : 385.0790. 
     Example 33 
     10-Chloro-6-((4-methylpentan-2-yl)amino)-12H-thiochromeno[2,3-c]quinolin-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%) (R f =0.9 at CH 2 Cl 2 ). Mp 176-177° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 0.98 (3H, d, J=6.8 Hz, —CH 3 ), 1.03 (3H, d, J=6.8 Hz, —CH 3 ), 1.35 (3H, d, J=6.4 Hz, —CH 3 ), 1.45 (1H, quin, J=6.4 Hz, —CH 2 —), 1.66 (1H, quin, J=6.8 Hz, —CH 2 —), 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) m/z calcd for C 22 H 19 N 2 OSCl [M] + : 396.1063. found [M+H] + : 397.1142. 
     Example 34 
     6-((2-Aminoethyl)amino)-10-chloro-12H-thiochromeno[2,3-c]quinolin-12-one (N14) 
     Product N14 was prepared from 3 and 1,2-diaminoethane. The pure compound was obtained as a yellow solid (yield 90%) (R f =0.6 at EA:MeOH:ammonia water=10:5:1). Mp 193-194° C. (MeOH).  1 H NMR (400 MHz, DMSO-d 6 ): δ (ppm) 2.90 (2H, t, J=6.0 Hz, —CH 2 —), 3.59 (2H, t, J=6.0 Hz, —CH 2 —), 7.36 (1H, t, J=8.0 Hz, Ar—H), 7.59 (1H, t, J=8.0 Hz, Ar—H), 7.66 (1H, 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).  13 C NMR (100 MHz, DMSO-d 6 ): δ (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 ( C O). HRMS (ESI) m/z calcd for C 18 H 14 N 3 OSCl [M] + : 355.0546. found [M+H] + : 356.0641. 
     Example 35 
     10-Chloro-6-((2-((2-hydroxyethyl)amino)ethyl)amino)-12H-thiochromeno[2,3-c]quinolin-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%) (R f =0.63 at EA:MeOH:ammonia water=10:5:1). Mp 141-143° C. (MeOH).  1 H NMR (400 MHz, DMSO-d 6 ): δ (ppm) 2.69 (2H, t, J=5.6 Hz, —CH 2 —), 2.90 (2H, t, J=6.0 Hz, —CH 2 —), 3.51 (2H, t, J=5.6 Hz, —CH 2 —), 3.65 (2H, t, J=6.0 Hz, —CH 2 —), 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).  13 C NMR (100 MHz, DMSO-d 6 ): δ (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 ( C O). HRMS (ESI) m/z calcd for C 20 H 18 N 3 O 2 SCl [M] + : 399.8938. found [M+H] + : 400.0880. 
     Example 36 
     10-Chloro-6-((2-morpholinoethyl)amino)-12H-thiochromeno[2,3-c]quinolin-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%) (R f =0.48 at EA). Mp 189-190° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 2.63 (4H, br, —CH 2 —), 2.81 (2H, br, —CH 2 —), 3.81 (6H, br, —CH 2 —), 5.92 (1H, 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) m/z calcd for C 22 H 20 N 3 O 2 SCl [M] + : 425.0965. found [M+H] + : 426.1058, [M−H] − : 424.0885. 
     Example 37 
     10-Chloro-6-((3-(dimethylamino)propyl)amino)-12H-thiochromeno[2,3-c]quinolin-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%) (R f =0.71 at EA:MeOH:ammonia water=10:5:1). Mp 194-195° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 1.92 (2H, quin, J=6.0 Hz, —CH 2 —), 2.41 (6H, s, —N(CH 3 ) 2 ), 2.60 (2H, t, J=5.6 Hz, —CH 2 N—), 3.81 (2H, q, J=5.6 Hz, —NCH 2 —), 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) m/z calcd for C 21 H 20 N 3 OSCl [M] + : 397.1016. found [M+H] + : 398.1072. 
     Example 38 
     10-Chloro-6-((3-(diethylamino)propyl)amino)-12H-thiochromeno[2,3-c]quinolin-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%) (R f =0.68 at EA:MeOH:ammonia water=10:5:1). Mp 142-143° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 1.15 (6H, t, J=6.8 Hz, —CH 3 ), 1.91 (2H, quin, J=6.0 Hz, —CH 2 —), 2.66-2.72 (6H, m, —NCH 2 —), 3.81 (2H, q, J=4.8 Hz, —NCH 2 —), 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) m/z calcd for C 23 H 24 N 3 SOCl [M] + : 425.1329. found [M+H] + : 426.1396, [M−H] − : 424.1284. 
     Example 39 
     10-Chloro-6-((3-((2-hydroxyethyl)amino)propyl)amino)-12H-thiochromeno[2,3-c]quinolin-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%) (R f =0.65 at EA:MeOH:ammonia water=10:5:1). Mp 65-67° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 1.89 (2H, quin, J=6.0 Hz, —CH 2 —), 2.15 (2H, br, —OH &amp; —NH—), 2.85 (4H, quin, —CH 2 —), 3.74 (2H, t, J=6.0 Hz, —CH 2 —), 3.80 (2H, t, J=5.2 Hz, —CH 2 —), 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 (1H, 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) m/z calcd for C 21 H 20 N 3 O 2 SCl [M] + : 413.0965. found [M+H] + : 414.1053, [M+H+2] + : 416.1037. 
     Example 40 
     10-Chloro-6-((2,3-dihydro-1H-inden-2-yl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one (N20) 
     Product N20 was prepared from 3 and 2-aminoindane. The pure compound was obtained as a brown solid (yield 65%) (R f =0.7 at CH 2 Cl 2 :n-hexane=2:1). Mp 251-252° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (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=5.2 Hz, indane-H), 7.21-7.25 (2H, 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).  13 C NMR (100 MHz, CDCl 3 ): δ (ppm) 40.41, 53.27, 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 ( C O). HRMS (ESI) m/z calcd for C 25 H 17 N 2 OSCl [M] + : 428.0750. found [M+H] + : 429.0822. 
     Example 41 
     10-Chloro-6-(cyclohexylamino)-12H-thiochromeno[2,3-c]quinolin-12-one (N21) 
     Product N21 was prepared from 3 and cyclohexylamine. The pure compound was obtained as a brown solid (yield 91%) (R f =0.7 at CH 2 Cl 2 :n-hexane=2:1). Mp 196-197° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 1.25-1.40 (4H, m, cyclohexylamine-CH 2 ), 1.49-1.60 (2H, m, cyclohexylamine-CH 2 ), 1.70-1.74 (2H, m, cyclohexylamine-CH 2 ), 1.79-1.84 (2H, m, cyclohexylamine-CH 2 ), 2.21 (2H, dd, J=8.8, 3.2 Hz, cyclohexylamine-CH 2 ), 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) m/z calcd for C 22 H 19 N 2 OSCl [M] + : 394.0907. found [M+H] + : 395.0991. 
     Example 42 
     6-((1-Benzylpiperidin-4-yl)amino)-10-chloro-12H-thiochromeno[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%) (R f =0.77 at EA). Mp 194-196° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (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 (2H, d, J=11.6 Hz, piperidine-H), 3.59 (2H, s, —CH 2 —), 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) m/z calcd for C 28 H 24 N 3 OSCl [M] + : 485.1329. found [M+H] + : 486.1379. 
     Example 43 
     10-Chloro-6-((thiophen-2-ylmethyl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one (N23) 
     Product N23 was prepared from 3 and 2-thiophenemethylamine. The pure compound was obtained as a brown solid (yield 78%) (R f =0.7 at CH 2 Cl 2 :n-hexane=2:1). Mp 178-180° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 5.07 (1H, d, J=5.2 Hz, —NCH 2 —), 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) m/z calcd for C 21 H 13 N 2 OS 2 Cl [M] + : 408.0158. found [M+H] + : 409.0251, [M−H] − : 407.0085. 
     Example 44 
     10-Chloro-6-((cyclohexylmethyl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one (N24) 
     Product N24 was prepared from 3 and cyclohexylmethanamine. The pure compound was obtained as a brown solid (yield 79%) (R f =0.7 at CH 2 Cl 2 :n-hexane=2:1). Mp 165-166° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 1.07 (1H, d, J=11.2 Hz, cyclohexyl-CH 2 ), 1.30 (1H, d, J=11.2 Hz, cyclohexyl-CH 2 ), 1.23 (2H, q, J=11.6 Hz, cyclohexyl-CH 2 ), 1.31 (2H, q, J=11.6 Hz, cyclohexyl-CH 2 ), 1.78-1.81 (4H, m, cyclohexyl-CH 2 ), 1.90 (2H, d, J=12.4 Hz, cyclohexyl-CH 2 ), 3.53 (2H, t, J=6.0 Hz, —NCH 2 —), 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) m/z calcd for C 23 H 21 N 2 OCl [M] + : 408.1063. found [M+H] + : 409.1115. 
     Example 45 
     6-(Benzylamino)-10-chloro-12H-thiochromeno[2,3-c]quinolin-12-one (N25) 
     Product N25 was prepared from 3 and benzylamine. The pure compound was obtained as a brown solid (yield 93%) (R f =0.67 at CH 2 Cl 2 :n-hexane=2:1). Mp 194-195° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 4.94 (2H, d, J=5.2 Hz, —CH 2 —), 5.16 (1H, br, —NH—), 7.33-7.51 (6H, 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) m/z calcd for C 23 H 15 N 2 OSCl [M] + : 402.0594. found [M+H] + : 403.0692. 
     Example 46 
     10-Chloro-6-((pyridin-2-ylmethyl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one (N26) 
     Product N26 was prepared from 3 and 2-picolylamine. The pure compound was obtained as a brown solid (yield 93%) (R f =0.25 at EA). Mp 187-189° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 5.01 (2H, d, J=4.0 Hz, —CH 2 —), 6.79 (1H, br, —NH—), 7.24-7.28 (1H, m, Ar′—H), 7.45 (2H, t, J=7.2 Hz, Ar′—H &amp; 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) m/z calcd for C 22 H 14 N 3 OSCl [M] + : 403.0546. found [M+H] + : 404.0615. 
     Example 47 
     6-((Benzo[d][1,3]dioxol-5-ylmethyl)amino)-10-chloro-12H-thiochromeno[2,3-c]quinolin-12-one (N27) 
     Product N27 was prepared from 3 and piperonylamine. The pure compound was obtained as a brown solid (yield 90%) (R f =0.88 at EA). Mp 205-206° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 4.82 (2H, t, J=5.2 Hz, —NCH 2 —), 5.08 (1H, br, —NH—), 5.97 (2H, s, —OCH 2 O—), 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 (2H, m, Ar—H), 7.86 (1H, 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) m/z calcd for C 24 H 15 N 2 O 3 SCl [M] + : 446.0492. found [M+H] + : 447.0586, [M−H] − : 445.0440. 
     Example 48 
     10-Chloro-6-((2-methoxybenzyl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one (N28) 
     Product N28 was prepared from 3 and 2-methoxybenzylamine. The pure compound was obtained as a brown solid (yield 82%) (R f =0.65 at CH 2 Cl 2 :n-hexane=2:1). Mp 223-224° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 3.95 (3H, s, —OCH 3 ), 4.93 (2H, d, J=5.6 Hz, —NCH 2 —), 5.57 (1H, t, J=5.6 Hz, —NH—), 5.97 (2H, s, —OCH 2 O—), 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).  13 C NMR (100 MHz, CDCl 3 ): δ (ppm) 42.33, 55.49, 110.47, 120.68, 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 ( C O). HRMS (ESI) m/z calcd for C 24 H 17 N 2 O 2 SCl [M] + : 432.0699. found [M+H] + : 433.0783. 
     Example 49 
     10-Chloro-6-((3,4-dimethoxybenzyl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one (N29) 
     Product N29 was prepared from 3 and 3,4-dimethoxybenzylamine. The pure compound was obtained as a brown solid (yield 84%) (R f =0.66 at CH 2 Cl 2 :n-hexane=2:1). Mp 251-252° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 3.89 (3H, s, —OCH 3 ), 3.90 (3H, s, —OCH 3 ), 4.86 (2H, d, J=4.8 Hz, —NCH 2 —), 5.11 (1H, t, J=5.2 Hz, —NH—), 6.89 (1H, d, J=8.0 Hz, Ar′—H), 7.05 (1H, 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) n/z calcd for C 25 H 19 N 2 O 3 SCl [M] + : 462.0805. found [M+H] + : 463.0900, [M−H] − : 461.0754. 
     Example 50 
     10-Chloro-6-(phenethylamino)-12H-thiochromeno[2,3-c]quinolin-12-one (N30) 
     Product N30 was prepared from 3 and phenethylamine. The pure compound was obtained as a brown solid (yield 94%) (R f =0.52 at CH 2 Cl 2 :n-hexane=2:1). Mp 151-152° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 3.10 (2H, t, J=6.8 Hz, —CH 2 —), 3.98 (2H, q, J=6.4 Hz, —NCH 2 —), 4.91 (1H, t, J=4.8 Hz, —NH—), 7.27-7.39 (5H, 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).  13 C NMR (100 MHz, CDCl 3 ): δ (ppm) 35.33, 43.52, 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 ( C O). HRMS (ESI) m/z calcd for C 24 H 17 N 2 OSCl [M] + : 416.9226. found [M+H] + : 417.0857, [M+H+2] + : 419.0834. 
     Example 51 
     10-Chloro-6-((4-methoxyphenethyl)amino)-12H-thiochromeno[2,3-c]quinolin-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%) (R f =0.89 at CH 2 Cl 2 ). Mp 214-215° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 3.03 (2H, t, J=6.8 Hz, —CH 2 —), 3.81 (3H, s, —OCH 3 ), 3.94 (2H, q, J=6.4 Hz, —NCH 2 —), 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) m/z calcd for C 25 H 19 N 2 O 2 SCl [M] + : 446.0856. found [M+H] + : 447.0938. 
     Example 52 
     6-((4-Aminophenethyl)amino)-10-chloro-12H-thiochromeno[2,3c]quinolin-12-one (N32) 
     Product N32 was prepared from 3 and 2-(4-aminophenyl)ethylamine. The pure compound was obtained as a yellow solid (yield 82%) (R f =0.52 at CH 2 Cl 2 ). Mp 208-210° C. (MeOH).  1 H NMR (400 MHz, CDCl 3 ): δ (ppm) 2.97 (2H, t, J=6.8 Hz, —CH 2 —), 3.63 (2H, br, —NH 2 ), 3.91 (2H, q, J=6.4 Hz, —NCH 2 —), 4.91 (1H, t, J=4.8 Hz, —NH—), 6.70 (2H, d, J=8.4 Hz, Ar′—H), 7.09 (2H, d, 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 (1H, 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).  13 C NMR (100 MHz, CDCl 3 ): δ (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 ( C O). HRMS (ESI) m/z calcd for C 24 H 16 N 3 OSCl [M] + : 431.0859. found [M+H] + : 432.0950. 
     Example 53 
     2-(10-Chloro-12-oxo-12H-thiochromeno[2,3-c]quinolin-6-yl)guanidine (TC-SCl-A-41) (N33) 
     Product N33 was a yellow solid (yield 85%). Mp: 370° C. (dec.)  1 H NMR (400 MHz, DMSO-d 6 ): δ ppm. 7.40 (3H, td, J=8.4, 1.2 Hz, Ar—H &amp; —NH 2 ), 7.59 (1H, td, J=8.7, 1.2 Hz, Ar—H), 7.59 (1H, 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=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).  13 C NMR (100 MHz, DMSO-d 6 ): δ 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 C 17 H 11 N 4 OSCl [M] + 354.0342. found [M+H] +  355.0438. 
     Example 54 
     10-Chloro-6-(piperidin-1-ylamino)-12H-thiochromeno[2,3-c]quinolin-12-one (TC-SCl-A-26) (N34) 
     Product N34 was a yellow solid (yield 60%). Mp: 180-181° C.  1 H NMR (400 MHz, CDCl 3 ): δ ppm. 1.72-1.74 (2H, m, —CH 2 —), 1.89 (4H, quin, J=5.2 Hz, —CH 2 —), 3.32 (4H, J=4.8 Hz, —CH 2 —), 7.36 (1H, tt, J=8.7, 2.1 Hz, Ar—H 10 ), 7.47 (1H, dd, J=8.4, 2.7 Hz, Ar—H 8 ), 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).  13 C NMR (100 MHz, CDCl 3 ): δ 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-81™, ATCC, Rockville, Md.) 125  and Bioresource Collection and Research Center (60122, BCRC, Taiwan) 126 , 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 2×10 3  cells were seeded into each well of a 96-well plate and incubated in 5% CO 2  at 37° C. for 24 h. To evaluate the in vitro cytotoxicity, all the synthetic compounds were dissolved in 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 μM). After 72 h, an amount of 100 μL of MTT (1 mg/mL) was added to each well, and the samples were incubated at 37° C. for 4 h. After removing the MTT solution, 100 μL 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 independent experiments. The IC 50  values of testing compounds were described in Table 1. 
     
       
         
               
             
               
               
               
             
               
               
               
               
             
           
               
                 TABLE 1 
               
             
             
               
                   
               
               
                 Effects of 10-Chloro-12H-thiochromeno[2,3-c]quinolin-12-one derivatives on 
               
               
                 cytotoxicity by MTT assay. 
               
             
          
           
               
                   
                   
                 MTT assay (IC 50  ± SD)  a   
               
             
          
           
               
                 No. 
                 R substitutions 
                 DU-145 (μM) 
                 PC-3 (μM) 
               
               
                   
               
               
                  2 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 &gt;15 
                 &gt;15 
               
               
                   
               
               
                  3 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 &gt;15 
                 &gt;15 
               
               
                   
               
               
                  4 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 &gt;15 
                 10.10 ± 1.81  
               
               
                   
               
               
                  5 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 10.84 ± 6.55  
                 3.89 ± 0.54 
               
               
                   
               
               
                  6 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 &gt;15 
                 &gt;15 
               
               
                   
               
               
                  7 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 5.01 ± 1.68 
                 2.84 ± 0.64 
               
               
                   
               
               
                  8 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 12.94 ± 0.26  
                 7.18 ± 2.45 
               
               
                   
               
               
                  9 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 &gt;15 
                 12.04 ± 3.41  
               
               
                   
               
               
                 10 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 &gt;15 
                 &gt;15 
               
               
                   
               
               
                 11 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 &gt;15 
                 &gt;15 
               
               
                   
               
               
                 12 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 &gt;15 
                 &gt;15 
               
               
                   
               
               
                 13 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 &gt;15 
                 &gt;15 
               
               
                   
               
               
                 14 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 &gt;15 
                 &gt;15 
               
               
                   
               
               
                 15 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 13.38 ± 2.87  
                 &gt;15 
               
               
                   
               
               
                 16 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 11.12 ± 4.18  
                 9.55 ± 2.42 
               
               
                   
               
               
                 17 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 &gt;15 
                 &gt;15 
               
               
                   
               
               
                 18 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 10.23 ± 2.03  
                 14.16 ± 1.41  
               
               
                   
               
               
                 19 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 9.02 ± 1.20 
                 6.36 ± 0.17 
               
               
                   
               
               
                 20 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 &gt;15 
                 &gt;15 
               
               
                   
               
               
                 21 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 11.12 ± 4.18  
                 9.55 ± 2.42 
               
               
                   
               
               
                 N1 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 11.65 ± 5.15  
                 14.48 ± 2.70  
               
               
                   
               
               
                 N2 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 6.70 ± 1.62 
                 5.18 ± 2.20 
               
               
                   
               
               
                 N3 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 7.64 ± 2.74 
                 7.41 ± 3.43 
               
               
                   
               
               
                 N4 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 10.35 ± 2.46  
                 10.11 ± 1.32  
               
               
                   
               
               
                 N5 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 &gt;15 
                 &gt;15 
               
               
                   
               
               
                 N6 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 &gt;15 
                 &gt;15 
               
               
                   
               
               
                 N7 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 5.94 ± 3.45 
                 2.25 ± 0.61 
               
               
                   
               
               
                 N8 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 1.73 ± 0.72 
                 1.11 ± 0.69 
               
               
                   
               
               
                 N9 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 11.55 ± 7.23  
                 7.48 ± 3.35 
               
               
                   
               
               
                 N10 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 5.94 ± 2.03 
                 6.31 ± 4.11 
               
               
                   
               
               
                 N11 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 4.95 ± 0.58 
                 8.38 ± 3.39 
               
               
                   
               
               
                 N12 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 6.74 ± 2.14 
                 7.05 ± 2.48 
               
               
                   
               
               
                 N13 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 14.16 ± 1.18  
                 10.38 ± 2.30  
               
               
                   
               
               
                 N14 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 1.80 ± 0.40 
                 1.64 ± 0.55 
               
               
                   
               
               
                 N15 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 3.81 ± 1.88 
                 3.44 ± 1.81 
               
               
                   
               
               
                 N16 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 &gt;15 
                 &gt;15 
               
               
                   
               
               
                 N17 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 2.70 ± 0.16 
                 2.27 ± 0.09 
               
               
                   
               
               
                 N18 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 3.53 ± 1.05 
                 2.22 ± 0.21 
               
               
                   
               
               
                 N19 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 1.50 ± 1.32 
                 1.98 ± 0.72 
               
               
                   
               
               
                 N20 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 4.86 ± 0.99 
                 6.20 ± 2.52 
               
               
                   
               
               
                 N21 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 8.06 ± 2.08 
                 7.51 ± 0.22 
               
               
                   
               
               
                 N22 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 8.39 ± 1.84 
                 6.47 ± 0.30 
               
               
                   
               
               
                 N23 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 14.65 ± 3.84  
                 &gt;15 
               
               
                   
               
               
                 N24 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 13.55 ± 3.88  
                 14.80 ± 3.26  
               
               
                   
               
               
                 N25  
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 13.49 ± 0.69 
                 11.16 ± 1.55  
               
               
                   
               
               
                 N26 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 14.75 ± 1.99  
                 14.20 ± 2.22  
               
               
                   
               
               
                 N27 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 11.82 ± 5.83  
                 &gt;15 
               
               
                   
               
               
                 N28 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 9.61 ± 4.88 
                 6.01 ± 3.31 
               
               
                   
               
               
                 N29 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 4.86 ± 0.99 
                 6.20 ± 2.52 
               
               
                   
               
               
                 N30 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 &gt;15 
                 &gt;15 
               
               
                   
               
               
                 N31 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 &gt;15 
                 &gt;15 
               
               
                   
               
               
                 N32 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 13.59 ± 1.39  
                 10.11 ± 0.94  
               
               
                   
               
               
                 N33 
                 
                   
                             
                     
                         
                         
                     
                   
                 
                 8.11 ± 1.37 
                 6.13 ± 6.98 
               
               
                   
               
               
                 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 μM ( 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 ( FIG. 4 ). The IC 50  value of compounds 7, N7, N14, N15, N18, N19, and N25 were about 10, 10, 1, 5, 25, 5, and 25 μM, 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 ( FIG. 7 ). The IC 50  value of compounds 7, N7, N8, N14, N15, N18, and N19 were about 10, 10, 1, 10, 5, 1, and 1 μM, 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)&#39;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 μM 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 μM) (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 a   
               
             
          
           
               
                   
                 2 
                 3 
                 4 
                 5 
                 6 
                 8 
                 10 
                 11 
                 12 
                 13 
               
               
                 Panel/ 
                 NSC- 
                 NSC- 
                 NSC- 
                 NSC- 
                 NSC- 
                 NSC- 
                 NSC- 
                 NSC- 
                 NSC- 
                 NSC- 
               
               
                 Cell line 
                 763977 
                 771780 
                 777199 
                 777200 
                 777202 
                 771775 
                 771776 
                 771777 
                 771778 
                 771779 
               
               
                   
               
             
          
           
               
                 Leukemia 
               
             
          
           
               
                 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 
                 93.81 
                 100.96 
                 113.79 
                 93.38 
               
               
                 K-562 
                 112.29 
                 101.04 
                 107.08 
                 95.83 
                 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 
                 70.01 
                 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 
                 89.02 
                 96.00 
                 88.18 
                 95.68 
                 86.41 
                 79.07 
                 85.15 
                 103.11 
                 88.91 
               
             
          
           
               
                 Non-small cell lung cancer 
               
             
          
           
               
                 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 
                 90.80 
                 96.64 
                 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-H322M 
                 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 
                 99.28 
               
               
                 NCI-H522 
                 81.64 
                 96.81 
                 86.60 
                 92.71 
                 70.42 
                 98.78 
                 94.76 
                 96.69 
                 93.50 
                 89.60 
               
             
          
           
               
                 Colon cancer 
               
             
          
           
               
                 COLO 205 
                 108.65 
                 112.15 
                 93.69 
                 97.94 
                 89.67 
                 90.06 
                 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-116 
                 103.87 
                 102.27 
                 86.20 
                 89.48 
                 74.26 
                 80.94 
                 69.33 
                 98.59 
                 105.36 
                 85.71 
               
               
                 HCT-15 
                 107.80 
                 102.28 
                 98.11 
                 107.99 
                 89.27 
                 93.94 
                 82.92 
                 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 
               
               
                 SW-620 
                 96.92 
                 99.06 
                 101.39 
                 103.05 
                 105.18 
                 95.50 
                 88.11 
                 93.08 
                 100.20 
                 96.18 
               
             
          
           
               
                 CNS cancer 
               
             
          
           
               
                 SF-268 
                 113.60 
                 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 
                 99.23 
                 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 
                 99.37 
               
               
                 SNB-75 
                 82.18 
                 77.88 
                 66.63 
                 52.28 
                 15.29 
                 42.15 
                 77.64 
                 82.62 
                 94.73 
                 76.88 
               
               
                 U251 
                 99.59 
                 101.71 
                 101.85 
                 99.18 
                 93.55 
                 92.52 
                 83.31 
                 96.69 
                 96.09 
                 98.58 
               
             
          
           
               
                 Melanoma 
               
             
          
           
               
                 LOX IMVI 
                 90.46 
                 90.53 
                 92.07 
                 91.21 
                 86.53 
                 87.47 
                 87.79 
                 94.22 
                 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 
                 102.05 
                 89.69 
                 105.86 
                 96.15 
                 87.80 
                 99.27 
                 91.36 
                 96.00 
                 104.48 
                 87.86 
               
             
          
           
               
                 Ovarian cancer 
               
             
          
           
               
                 IGROV1 
                 103.57 
                 96.06 
                 78.35 
                 71.44 
                 68.16 
                 80.08 
                 71.67 
                 93.49 
                 91.38 
                 80.90 
               
               
                 OVCAR-3 
                 121.29 
                 111.27 
                 106.37 
                 96.60 
                 79.65 
                 89.13 
                 93.92 
                 101.69 
                 109.63 
                 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 
                 112.47 
                 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 
               
               
                 NCI/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.T. b   
                 101.32 
                 94.40 
                 58.94 
                 41.57 
                 75.11 
                 95.12 
                 94.79 
                 100.39 
                 77.18 
               
             
          
           
               
                 Renal cancer 
               
             
          
           
               
                 786-0 
                 104.44 
                 105.25 
                 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 
                 N.T. 
                 N.T. 
                 N.T. 
                 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 
               
               
                 SN12C 
                 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 
               
               
                 UO-31 
                 68.91 
                 66.06 
                 76.44 
                 74.69 
                 73.81 
                 48.97 
                 42.34 
                 64.91 
                 63.85 
                 73.11 
               
             
          
           
               
                 Prostate cancer 
               
             
          
           
               
                 PC-3 
                 93.23 
                 91.59 
                 91.90 
                 92.46 
                 81.25 
                 84.81 
                 77.99 
                 89.12 
                 90.66 
                 79.80 
               
               
                 DU-145 
                 115.22 
                 114.27 
                 99.80 
                 104.27 
                 92.93 
                 103.45 
                 96.26 
                 111.61 
                 111.15 
                 110.81 
               
             
          
           
               
                 Breast cancer 
               
             
          
           
               
                 MCF 
                 91.95 
                 82.58 
                 69.90 
                 65.39 
                 22.19 
                 76.60 
                 51.73 
                 68.32 
                 74.16 
                 50.59 
               
               
                 MDA-MB-231/ 
                 95.70 
                 84.28 
                 81.80 
                 78.12 
                 71.03 
                 57.30 
                 55.49 
                 89.11 
                 94.34 
                 71.53 
               
               
                 ATCC 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 HS 578T 
                 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-47D 
                 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 
                 99.30 
                 90.52 
               
               
                 Delta 
                 33.20 
                 32.05 
                 30.54 
                 39.82 
                 65.30 
                 68.64 
                 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 a   
               
             
          
           
               
                   
                 N1 
                 N2 
                 N6 
                 N7 
                 N9 
                 N12 
                 N13 
                 N14 
               
               
                 Panel/ 
                 NSC- 
                 NSC- 
                 NSC- 
                 NSC- 
                 NSC- 
                 NSC- 
                 NSC- 
                 NSC- 
               
               
                 Cell line 
                 775512 
                 771784 
                 775513 
                 771785 
                 771786 
                 772854 
                 772855 
                 772865 
               
               
                   
               
             
          
           
               
                 Leukemia 
               
             
          
           
               
                 CCRF-CEM 
                 79.77 
                 88.62 
                 93.72 
                 −53.54 
                 81.07 
                 70.00 
                 98.57 
                 −39.61 
               
               
                 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 
                 −51.67 
               
             
          
           
               
                 Non-small cell lung cancer 
               
             
          
           
               
                 A549/ATCC 
                 51.68 
                 61.84 
                 91.04 
                 23.38 
                 80.72 
                 69.86 
                 101.98 
                 −88.76 
               
               
                 EKVX 
                 N.T. b   
                 N.T. 
                 N.T. 
                 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. 
               
               
                 NCI-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 
                 −62.39 
               
               
                 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 
                 −83.65 
                 101.55 
                 99.61 
                 112.26 
                 −90.43 
               
               
                 HCT-116 
                 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 
                 116.70 
                 −84.24 
                 87.36 
                 78.24 
                 121.02 
                 −83.30 
               
               
                 KM12 
                 68.75 
                 85.79 
                 94.04 
                 −77.07 
                 91.93 
                 82.37 
                 105.66 
                 −85.99 
               
               
                 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 
                 84.97 
                 80.67 
                 108.13 
                 −64.64 
                 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 
                 −76.79 
                 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 
                 −48.22 
                 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 
               
             
          
           
               
                 IGROV1 
                 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 
                 −52.16 
               
               
                 OVCAR-8 
                 31.61 
                 62.54 
                 95.31 
                 1.22 
                 78.13 
                 68.74 
                 92.79 
                 −74.59 
               
               
                 NCI/ADR-RES 
                 70.95 
                 88.61 
                 101.32 
                 23.00 
                 91.54 
                 77.02 
                 98.47 
                 −42.68 
               
               
                 SK-OV-3 
                 88.67 
                 28.90 
                 99.71 
                 70.10 
                 104.57 
                 58.93 
                 86.70 
                 12.25 
               
             
          
           
               
                 Renal cancer 
               
             
          
           
               
                 786-0 
                 95.51 
                 23.87 
                 108.84 
                 43.11 
                 104.43 
                 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 
               
               
                 CAKI-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 
               
               
                 SN12C 
                 92.52 
                 84.57 
                 94.28 
                 48.67 
                 92.52 
                 81.98 
                 95.14 
                 −93.98 
               
               
                 TK-10 
                 81.21 
                 47.11 
                 138.89 
                 −89.05 
                 78.14 
                 77.45 
                 123.23 
                 −28.04 
               
               
                 UO-31 
                 65.34 
                 32.06 
                 78.57 
                 21.83 
                 51.97 
                 51.01 
                 78.64 
                 −92.77 
               
             
          
           
               
                 Prostate cancer 
               
             
          
           
               
                 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/ 
                 66.58 
                 48.66 
                 77.83 
                 −37.28 
                 66.62 
                 58.29 
                 84.22 
                 −84.52 
               
               
                 ATCC 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 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 
                 26.29 
                 95.79 
                 12.81 
                 39.59 
                 80.85 
                 96.02 
                 −62.13 
               
               
                 Mean 
                 75.06 
                 65.97 
                 96.94 
                 −11.06 
                 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 
                 74.81 
                 187.88 
                 101.96 
                 68.60 
                 49.47 
                 176.87 
               
               
                   
               
               
                   a Data obtained from NCI in vitro 60-cell drug screen program at 10 −5  molar concentration. 
               
               
                   b 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/ 
                 N16 
                 N17 
                 N19 
                 N21 
                 N25 
                 N27 
                 N30 
                 N31 
               
               
                 Cell line 
                 NSC777203 
                 NSC775511 
                 NSC777201 
                 NSC772866 
                 NSC772867 
                 NSC775508 
                 NSC775509 
                 NSC775510 
               
               
                   
               
             
          
           
               
                 Leukemia 
               
             
          
           
               
                 CCRF-CEM 
                 82.21 
                 69.06 
                 −54.78 
                 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 
                 −64.45 
                 85.28 
                 68.33 
                 104.74 
                 88.44 
                 93.49 
               
               
                 EKVX 
                 N.T. b   
                 N.T. 
                 N.T. 
                 N.T. 
                 N.T. 
                 N.T. 
                 N.T. 
                 N.T. 
               
               
                 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 
                 10.05 
                 91.10 
                 91.00 
                 73.65 
                 117.72 
                 99.19 
               
               
                 NCI-H460 
                 100.49 
                 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 
                 98.46 
               
               
                 HCT-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 
                 120.78 
               
               
                 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 
                 102.93 
                 97.62 
                 100.6 
                 96.74 
               
               
                 SNB-75 
                 60.74 
                 60.99 
                 23.66 
                 81.91 
                 14.83 
                 57.14 
                 68.76 
                 74.50 
               
               
                 U251 
                 76.34 
                 84.19 
                 −66.76 
                 N.T. 
                 N.T. 
                 81.03 
                 96.78 
                 98.72 
               
             
          
           
               
                 Melanoma 
               
             
          
           
               
                 LOX IMVI 
                 67.43 
                 66.16 
                 −82.79 
                 N.T. 
                 N.T. 
                 78.69 
                 89.03 
                 87.78 
               
               
                 MALME-3M 
                 78.79 
                 100.91 
                 −34.30 
                 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 
               
               
                 SK-MEL-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. 
                 N.T. 
               
               
                 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 
                 102.03 
                 100.83 
               
               
                 OVCAR-8 
                 92.43 
                 98.38 
                 9.89 
                 74.91 
                 68.33 
                 79.45 
                 95.61 
                 97.83 
               
               
                 NCI/ADR-RES 
                 77.03 
                 96.94 
                 26.35 
                 95.91 
                 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 
               
             
          
           
               
                 Renal 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 
                 58.48 
                 90.85 
                 91.19 
               
               
                 CAKI-1 
                 N.T. 
                 84.31 
                 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 
                 87.80 
                 85.53 
                 86.54 
                 94.23 
               
               
                 TK-10 
                 76.67 
                 108.89 
                 −92.61 
                 127.96 
                 75.80 
                 116.79 
                 122.13 
                 132.57 
               
               
                 UO-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 
                 104.25 
                 74.80 
                 72.44 
                 100.17 
                 107.03 
               
             
          
           
               
                 Breast cancer 
               
             
          
           
               
                 MCF7 
                 51.69 
                 51.20 
                 −72.49 
                 46.14 
                 16.01 
                 43.53 
                 60.91 
                 59.13 
               
               
                 MDA-MB-231/ 
                 59.20 
                 73.39 
                 −66.53 
                 77.17 
                 32.67 
                 54.65 
                 79.55 
                 82.31 
               
               
                 ATCC 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 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 
                 81.18 
                 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 
               
               
                   
               
               
                   a Data obtained from NCI in vitro 60-cell drug screen program at 10 −5  molar concentration. 
               
               
                   b N.T. = No test. 
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 5 
               
             
             
               
                   
               
               
                 In vitro antitumor activity (GI 50  in μM), toxicity (LC 50  in 
               
               
                 μM) and TGI data of selected compounds N2, N7, N14, N19, and N25. 
               
             
          
           
               
                 Panel/ 
                 N2 
                 N7 
                 N14 
                 N19 
                 N25 
               
               
                 Cell line 
                 (NSC771784) 
                 (NSC771785) 
                 (NSC772865) 
                 (NSC777201) 
                 (NSC772867) 
               
             
          
           
               
                 (μM) 
                 GI 50   
                 TGI 
                 LC 50   
                 GI 50   
                 TGI 
                 LC 50   
                 GI 50   
                 TGI 
                 LC 50   
                 GI 50   
                 TGI 
                 LC 50   
                 GI 50   
                 TGI 
                 LC 50   
               
               
                   
               
             
          
           
               
                 Leukemia 
               
             
          
           
               
                 CCRF- 
                 &gt;100 
                 &gt;100 
                 &gt;100 
                 1.63 
                 3.41 
                  7.15 
                 1.75 
                 3.65 
                  7.64 
                 1.83 
                 3.80 
                 — 
                 &gt;100 
                 &gt;100 
                 &gt;100 
               
               
                 CEM 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 HL-60 
                 &gt;100 
                 &gt;100 
                 &gt;100 
                 1.76 
                 3.49 
                  6.95 
                 N.T. 
                 N.T. 
                 N.T. 
                 N.T. 
                 N.T. 
                 N.T. 
                 N.T. 
                 N.T. 
                 N.T. 
               
               
                 (TB) 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 K-562 
                 &gt;100 
                 &gt;100 
                 &gt;100 
                 1.33 
                 3.00 
                  6.74 
                 1.69 
                 3.65 
                  7.84 
                 1.73 
                 — 
                 &gt;100 
                 &gt;100 
                 &gt;100 
                 &gt;100 
               
               
                 MOLT-4 
                 &gt;100 
                 &gt;100 
                 &gt;100 
                 1.71 
                 3.40 
                  6.74 
                 2.02 
                 3.96 
                  7.77 
                 1.88 
                 — 
                 — 
                 &gt;100 
                 &gt;100 
                 &gt;100 
               
               
                 RPMI- 
                 &gt;100 
                 &gt;100 
                 &gt;100 
                 1.76 
                 3.68 
                  7.69 
                 2.26 
                 4.92 
                 62.30 
                 1.84 
                 4.21 
                 — 
                 &gt;100 
                 &gt;100 
                 &gt;100 
               
               
                 8226 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 SR 
                 &gt;100 
                 &gt;100 
                 &gt;100 
                 1.70 
                 3.73 
                  8.14 
                 1.81 
                 4.30 
                 &gt;100 
                 1.98 
                 4.55 
                 &gt;100 
                 &gt;100 
                 &gt;100 
                 &gt;100 
               
             
          
           
               
                 Non-small cell lung cancer 
               
             
          
           
               
                 A549/ 
                 48.1 
                 &gt;100 
                 &gt;100 
                 1.66 
                 3.17 
                  6.06 
                 1.83 
                 3.50 
                  6.68 
                 1.76 
                 3.30 
                 — 
                 14.40 
                 &gt;100 
                 &gt;100 
               
               
                 ATCC 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 EKVX 
                 N.T. 
                 N.T. 
                 N.T. 
                 N.T. 
                 N.T. 
                 N.T. 
                 N.T. 
                 N.T. 
                 N.T. 
                 N.T. 
                 N.T. 
                 N.T. 
                 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. 
                 N.T. 
                 N.T. 
                 1.03 
                 3.05 
                  9.04 
                 1.23 
                 4.58 
                 26.70 
                  1.75 
                  7.53 
                 53.60 
               
               
                 NCI-H226 
                 33.20 
                 &gt;100 
                 &gt;100 
                 1.84 
                 4.38 
                 24.30 
                 1.36 
                 2.95 
                 — 
                 1.90 
                 4.46 
                 &gt;100 
                  1.84 
                 — 
                 &gt;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 
                 &gt;100 
                 &gt;100 
               
               
                 NCI- 
                 69.50 
                 &gt;100 
                 &gt;100 
                 1.74 
                 3.18 
                  5.81 
                 1.65 
                 3.34 
                  6.74 
                 1.80 
                 3.56 
                  7.03 
                 &gt;100 
                 &gt;100 
                 &gt;100 
               
               
                 H322M 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 NCI-H460 
                 &gt;100 
                 &gt;100 
                 &gt;100 
                 1.76 
                 3.38 
                  6.50 
                 1.69 
                 3.45 
                  7.05 
                 1.66 
                 3.35 
                 — 
                  5.00 
                 &gt;100 
                 &gt;100 
               
               
                 NCI-H522 
                 24.80 
                 &gt;100 
                 &gt;100 
                 1.59 
                 3.07 
                  5.90 
                 1.86 
                 3.59 
                  6.92 
                 1.85 
                 3.56 
                 — 
                 25.30 
                 &gt;100 
                 &gt;100 
               
             
          
           
               
                 Colon cancer 
               
             
          
           
               
                 COLO 205 
                 &gt;100 
                 &gt;100 
                 &gt;100 
                 1.66 
                 3.17 
                  6.05 
                 1.73 
                 3.57 
                 — 
                 1.63 
                 3.37 
                 — 
                 — 
                 &gt;100 
                 &gt;100 
               
               
                 HCC-2998 
                 &gt;100 
                 &gt;100 
                 &gt;100 
                 1.68 
                 3.38 
                  6.82 
                 2.03 
                 4.06 
                  8.14 
                 1.58 
                 3.42 
                 — 
                 &gt;100 
                 &gt;100 
                 &gt;100 
               
               
                 HCT-116 
                  6.31 
                 &gt;100 
                 &gt;100 
                 1.57 
                 3.18 
                  6.45 
                 1.63 
                 2.99 
                  5.47 
                 1.60 
                 3.05 
                 — 
                  5.59 
                 &gt;100 
                 &gt;100 
               
               
                 HCT-15 
                 &gt;100 
                 &gt;100 
                 &gt;100 
                 1.59 
                 3.14 
                  6.21 
                 1.58 
                 3.12 
                  6.16 
                 1.61 
                 3.25 
                  6.57 
                  8.75 
                 &gt;100 
                 &gt;100 
               
               
                 HT29 
                 &gt;100 
                 &gt;100 
                 &gt;100 
                 1.65 
                 3.36 
                  6.88 
                 1.78 
                 3.29 
                  6.09 
                 2.05 
                 4.00 
                  7.82 
                 — 
                 &gt;100 
                 &gt;100 
               
               
                 KM12 
                 &gt;100 
                 &gt;100 
                 &gt;100 
                 1.78 
                 3.28 
                  6.03 
                 1.67 
                 3.11 
                  5.79 
                 1.67 
                 3.31 
                 — 
                 48.30 
                 &gt;100 
                 &gt;100 
               
               
                 SW-620 
                 &gt;100 
                 &gt;100 
                 &gt;100 
                 1.71 
                 3.29 
                  6.36 
                 1.67 
                 3.42 
                  7.02 
                 1.63 
                 3.30 
                 — 
                 — 
                 &gt;100 
                 &gt;100 
               
             
          
           
               
                 CNS cancer 
               
             
          
           
               
                 SF-268 
                 22.10 
                 &gt;100 
                 &gt;100 
                 1.57 
                 3.16 
                  6.35 
                 1.59 
                 3.23 
                  6.55 
                 1.67 
                 3.35 
                 — 
                 13.80 
                 &gt;100 
                 &gt;100 
               
               
                 SF-295 
                 12.20 
                 35.20 
                 &gt;100 
                 1.73 
                 3.21 
                  5.98 
                 1.79 
                 3.63 
                 — 
                 1.73 
                 3.33 
                 — 
                  3.50 
                 16.50 
                 &gt;100 
               
               
                 SF-539 
                 10.10 
                 58.50 
                 &gt;100 
                 1.68 
                 3.08 
                  5.67 
                 1.57 
                 2.95 
                  5.52 
                 1.70 
                 3.22 
                  6.11 
                  4.49 
                 46.40 
                 &gt;100 
               
               
                 SNB-19 
                 26.60 
                 81.10 
                 &gt;100 
                 1.54 
                 3.07 
                  6.10 
                 1.65 
                 3.18 
                 — 
                 1.64 
                 3.26 
                 — 
                  8.56 
                 &gt;100 
                 &gt;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 
                 &gt;100 
                 1.48 
                 2.84 
                  5.44 
                 1.63 
                 3.07 
                  5.78 
                 1.73 
                 3.22 
                 — 
                  4.41 
                 25.10 
                 &gt;100 
               
             
          
           
               
                 Melanoma 
               
             
          
           
               
                 LOX  
                 &gt;100 
                 &gt;100 
                 &gt;100 
                 1.68 
                 3.24 
                  6.24 
                 1.68 
                 3.16 
                  5.96 
                 1.80 
                 3.80 
                 — 
                 &gt;100 
                 &gt;100 
                 &gt;100 
               
               
                 IMVI 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 MALME- 
                 N.T. 
                 N.T. 
                 N.T. 
                 N.T. 
                 N.T. 
                 N.T. 
                 1.98 
                 3.96 
                  7.93 
                 2.15 
                 4.33 
                 — 
                 &gt;100 
                 &gt;100 
                 &gt;100 
               
               
                 3M 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 M14 
                 &gt;100 
                 &gt;100 
                 &gt;100 
                 1.85 
                 3.44 
                  6.40 
                 1.83 
                 3.35 
                  6.14 
                 1.83 
                 3.53 
                  6.82 
                 &gt;100 
                 &gt;100 
                 &gt;100 
               
               
                 MDA-MB- 
                 &gt;100 
                 &gt;100 
                 &gt;100 
                 1.71 
                 3.22 
                  6.05 
                 1.82 
                 3.59 
                 — 
                 1.59 
                 2.99 
                  5.62 
                 &gt;100 
                 &gt;100 
                 &gt;100 
               
               
                 435 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 SK-Mel-2 
                 16.3 
                 43.2 
                 &gt;100 
                 1.80 
                 3.32 
                  6.12 
                 1.98 
                 3.95 
                  7.87 
                 2.05 
                 3.77 
                  6.90 
                 10.60 
                 31.90 
                 96.50 
               
               
                 SK-MEL- 
                 &gt;100 
                 &gt;100 
                 &gt;100 
                 1.81 
                 3.27 
                  5.89 
                 1.75 
                 3.15 
                  5.66 
                 1.89 
                 3.56 
                 — 
                 — 
                 &gt;100 
                 &gt;100 
               
               
                 28 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 SK- 
                 — 
                 &gt;100 
                 &gt;100 
                 1.57 
                 3.59 
                  8.21 
                 1.51 
                 2.87 
                  5.44 
                 1.60 
                 3.00 
                  5.63 
                  4.23 
                 &gt;100 
                 &gt;100 
               
               
                 MEL-5 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 UACC- 
                 &gt;100 
                 &gt;100 
                 &gt;100 
                 1.74 
                 3.21 
                  5.93 
                 1.90 
                 3.56 
                  6.70 
                 1.86 
                 3.44 
                  6.33 
                 &gt;100 
                 &gt;100 
                 &gt;100 
               
               
                 257 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 UACC-62 
                 22.3 
                 &gt;100 
                 &gt;100 
                 1.70 
                 3.16 
                  5.88 
                 1.67 
                 3.30 
                  6.52 
                 1.81 
                 3.40 
                  6.39 
                 16.90 
                 &gt;100 
                 &gt;100 
               
             
          
           
               
                 Ovarian cancer 
               
             
          
           
               
                 IGROV1 
                 11.90 
                 54.40 
                 &gt;100 
                 1.70 
                 3.34 
                  6.58 
                 1.61 
                 3.45 
                 — 
                 1.66 
                 3.60 
                 — 
                  4.64 
                 &gt;100 
                 &gt;100 
               
               
                 OVCAR-3 
                 52.40 
                 &gt;100 
                 &gt;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 
                 &gt;100 
                 &gt;100 
                 &gt;100 
                 1.19 
                 2.75 
                  6.35 
                 1.52 
                 3.15 
                  6.54 
                 1.60 
                 3.23 
                 — 
                 15.00 
                  3.74 
                 — 
               
               
                 OVCAR-5 
                 &gt;100 
                 &gt;100 
                 &gt;100 
                 1.45 
                 2.91 
                  5.83 
                 1.46 
                 2.88 
                  5.66 
                 1.63 
                 3.50 
                  7.53 
                 &gt;100 
                 &gt;100 
                 &gt;100 
               
               
                 OVCAR-8 
                 17.70 
                 &gt;100 
                 &gt;100 
                 1.64 
                 3.15 
                  6.06 
                 1.89 
                 4.12 
                 — 
                 1.75 
                 3.32 
                 — 
                  7.75 
                 &gt;100 
                 &gt;100 
               
               
                 NCI/ADR- 
                 33.50 
                 &gt;100 
                 &gt;100 
                 1.67 
                 3.17 
                  6.00 
                 1.88 
                 3.67 
                 — 
                 1.82 
                 3.63 
                 — 
                 43.00 
                 &gt;100 
                 &gt;100 
               
               
                 RES 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 SK-OV-3 
                 11.10 
                 35.20 
                 &gt;100 
                 1.68 
                 3.06 
                  5.58 
                 1.78 
                 3.38 
                  6.42 
                 1.68 
                 3.38 
                  6.80 
                  3.56 
                 13.90 
                 &gt;100 
               
             
          
           
               
                 Renal cancer 
               
             
          
           
               
                 786-0 
                 10.5 
                 33.8 
                 &gt;100 
                 1.84 
                 3.45 
                  6.46 
                 1.76 
                 3.20 
                  5.81 
                 1.72 
                 3.37 
                 — 
                  9.55 
                 48.70 
                 &gt;100 
               
               
                 A498 
                 15.3 
                 91.3 
                 &gt;100 
                 2.70 
                 9.44 
                  3.52 
                 1.45 
                 4.99 
                 29.20 
                 1.27 
                 3.62 
                 15.00 
                  8.05 
                 39.20 
                 &gt;100 
               
               
                 ACHN 
                  8.66 
                 &gt;100 
                 &gt;100 
                 1.63 
                 3.07 
                  5.77 
                 1.44 
                 2.76 
                  5.29 
                 1.67 
                 3.22 
                 — 
                  4.02 
                 &gt;100 
                 &gt;100 
               
               
                 CAKI-1 
                 26.4 
                 &gt;100 
                 &gt;100 
                 1.43 
                 2.80 
                  5.49 
                 1.56 
                 3.27 
                 — 
                 1.33 
                 2.69 
                  5.45 
                  6.07 
                 &gt;100 
                 &gt;100 
               
               
                 RXF 393 
                 13.1 
                 44.6 
                 &gt;100 
                 1.60 
                 3.41 
                  7.29 
                 N.T. 
                 N.T. 
                 N.T. 
                 1.64 
                 3.20 
                 — 
                 N.T. 
                 N.T. 
                 N.T. 
               
               
                 SN12C 
                 75.3 
                 &gt;100 
                 &gt;100 
                 1.59 
                 3.12 
                  6.12 
                 1.45 
                 3.02 
                 — 
                 1.70 
                 3.39 
                  6.80 
                 24.90 
                 &gt;100 
                 &gt;100 
               
               
                 TK-10 
                 N.T. 
                 N.T. 
                 &gt;100 
                 N.T. 
                 N.T. 
                 N.T. 
                 2.06 
                 3.58 
                  6.21 
                 2.09 
                 3.74 
                  6.69 
                  5.02 
                 22.30 
                 &gt;100 
               
               
                 UO-31 
                 29.4 
                 &gt;100 
                 &gt;100 
                 1.19 
                 2.50 
                  5.22 
                 1.29 
                 2.78 
                  5.96 
                 1.36 
                 2.94 
                  6.35 
                 57.80 
                 &gt;100 
                 &gt;100 
               
             
          
           
               
                 Prostate cancer 
               
             
          
           
               
                 PC-3 
                 &gt;100 
                 &gt;100 
                 &gt;100 
                 1.47 
                 2.90 
                  5.74 
                 1.55 
                 2.99 
                  5.76 
                 1.51 
                 2.92 
                 5.66 
                 56.90 
                 &gt;100 
                 &gt;100 
               
               
                 DU-145 
                 &gt;100 
                 &gt;100 
                 &gt;100 
                 1.87 
                 3.34 
                  5.99 
                 1.81 
                 3.20 
                  5.66 
                 1.76 
                 3.31 
                 6.25 
                  5.50 
                 &gt;100 
                 &gt;100 
               
             
          
           
               
                 Breast cancer 
               
             
          
           
               
                 MCF7 
                  0.0.47 
                 &gt;100 
                 &gt;100 
                 1.23 
                 2.66 
                  5.71 
                 1.28 
                 2.81 
                 — 
                 1.05 
                 2.60 
                 — 
                  0.04 
                 18.60 
                 &gt;100 
               
               
                 MDA-MB- 
                 10.10 
                 65.5 
                 &gt;100 
                 1.20 
                 2.51 
                  5.28 
                 1.31 
                 2.65 
                  5.35 
                 1.36 
                 2.86 
                  6.02 
                  2.36 
                 34.80 
                 &gt;100 
               
               
                 231/ATCC 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 HS 578T 
                  9.64 
                 65.5 
                 &gt;100 
                 1.74 
                 4.02 
                  9.27 
                 1.69 
                 3.90 
                 — 
                 1.29 
                 3.53 
                  9.64 
                  2.94 
                 21.20 
                 &gt;100 
               
               
                 BT-549 
                  9.87 
                 &gt;100 
                 &gt;100 
                 8.91 
                 2.27 
                  5.33 
                 2.15 
                 5.35 
                 62.00 
                 1.67 
                 3.15 
                 — 
                 20.50 
                 &gt;100 
                 &gt;100 
               
               
                 T-47D 
                 N.T. 
                 N.T. 
                 N.T. 
                 N.T. 
                 N.T. 
                 N.T. 
                 1.56 
                 3.37 
                 — 
                 1.50 
                 3.43 
                 — 
                  1.26 
                  7.11 
                 &gt;100 
               
               
                 MDA-MB- 
                  0.040 
                 &gt;100 
                 &gt;100 
                 1.47 
                 3.28 
                  7.34 
                 1.27 
                 2.80 
                  6.15 
                 1.06 
                 2.56 
                  6.20 
                  0.03 
                  0.66 
                 63.10 
               
               
                 486 
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                 Mean 
                 28.84 
                 83.18 
                 100 
                 1.66 
                 3.39 
                  6.92 
                 1.66 
                 3.39 
                  7.94 
                 1.66 
                 3.39 
                  9.55 
                 11.22 
                 53.70 
                 91.20