TWI488843B - Novel thiochromeno(2,3-c)quinolin-12-one derivatives, preparation method and application thereof - Google Patents

Description

Novel dehydrothiamine [2,3-c]quinolin-12-one derivative, preparation method thereof and application thereof

The present invention relates to the technical field of cancer drug development, and in particular to dehydrothiamine [2,3- c ]quinolin-12-one derivatives, their anticancer activity application and evaluation.

In most organisms, the replication and maintenance of telomere length must be dependent on telomerase. Telomerase is a ribonuclease that is a complex of RNA and protein subunits. At present, some important telomerase subunits have been identified, including human telomerase reverse transcriptase (hTERT) with reverse transcriptase activity, and as a human telomerase composition. The human telomerase RNA component of the template, and some proteins that bind to telomerase, such as human telomerase-associated protein, p23, hsp90, hsp40, hsp70, and the like.

Many studies have pointed out that human telomerase activity exists only in some highly proliferating cells, such as germ cells, hematopoietic cells, part of stem cells, most immortalized cells, and Most of the tumor cells. In general somatic cells, since telomerase activity is not contained, telomeres are gradually reduced as the number of cell divisions increases, which can be regarded as a mitotic clock for cells to calculate the number of divisions. When the telomere is short to a certain extent, the cell will stop dividing and enter the aging stage, and the cell will stay at this stage for a period of time, and then go to death, this period is called the M1 phase (mortality stage 1); if in the M1 phase Mutations in the tumor suppressor gene, such as p53 and Rb, cause the cell to escape the aging phase and continue cell division, which is called the M2 phase (mortality stage 2). Due to the absence of telomerase activity during this period, the telomere length will still be shortened. When the telomere can not protect the integrity of the end of the chromosome, which leads to chromosomal instability, the cell can not completely transmit the genetic message and gradually die, so the M2 phase is also called the crisis; most of the cells will M2 phase death, only a few cells will survive due to the expression of telomerase activity, this cell can be unrestricted and persistently split into undead cells (or cancer cells).

Therefore, it is generally believed that activation of telomerase can maintain telomere length to prevent cells from entering replicative senescence, or to remove telomerase activity and inhibit its activity to limit the division of cancer cells, both of which can be cell-oriented and cancerous. The key to development. In summary, since normal human somatic cells do not have telomerase activity, a drug capable of inhibiting telomerase activity does not affect the physiological state of normal cells when inhibiting telomerase activity. However, this active inhibitory drug can attenuate the proliferative ability of cancer cells, so if telomerase activity inhibitors can be found, the development of cancer therapeutic drugs has great potential through the discussion of anticancer mechanisms.

The cause of cancer is due to abnormal proliferation of DNA. Therefore, as long as it can selectively attack the DNA of cancer cells, it is not desirable to damage the DNA of normal cells, but it is difficult to distinguish the DNA of normal cells from the DNA of cancer cells, so that normal cells and cancer cells are found. After the difference in characteristics, a specific "target treatment" is produced. This treatment, combined with other chemotherapy or radiation therapy, can significantly reduce side effects and achieve better results, so target treatment is now popular. Research to treat cancer in the direction. Because topoisomerase plays an indispensable role in DNA replication, it is the target of recent anti-cancer target treatment research.

Camptothecin is the first small molecule drug to inhibit the first type of topoisomerase, but unfortunately, camptothecin has many disadvantages that make it impossible to use in clinical medicine, such as lactone ring in the human body. It is easy to be hydrolyzed into hydroxycarboxylate under pH environment, which leads to easy to bind to serum albumin in vivo and loses the effect of inhibiting the first type of topoisomerase; the triple complex of camptothecin-Top I-DNA is not The structure of the covalent bond is unstable; the water solubility of camptothecin is poor, resulting in poor bioavailability; the p-glycoprotein (MDR1, ABCB1) on the cell membrane is discharged from the pump to push the drug out of the cell; More importantly, some tumor cells gradually develop resistance to camptothecin and its derivatives and adverse drug side effects, making it even more semi-synthetic than the camptothecin. The drug, in 1996, was approved by the US Food and Drug Administration (FDA) for Topotecan (Hycamtin ^® ) for the treatment of ovarian cancer and Irinotecan (Camptosar ^® ) for the treatment of colon cancer.

In view of the importance of the activity of the topoisomerase activity inhibitor in the development of cancer therapeutic drugs, the inventors of the present invention have finally developed and developed the dehydrothiamine [2,3- c ]quinolin-12-one derivative, Its preparation method and its application.

It is an object of the present invention to provide a compound of formula (I), Wherein R is selected from the group consisting of: i) halo, amine, hydroxy and thio; ii) linear alkyl of NH(CH ₂ ) _n H, alkyl having a branch substituent An alkyl branch having an amino group substitution, an alkyl group having a hydroxyl group substitution, wherein 0 n 12; iii) O(CH ₂ ) _n H, N(CH ₃ ) ₂ , NH(CH ₂ ) _n NH(CH ₂ ) _n OH,; n 10; iv) C _3-12 nitrogen-containing cycloalkyl and heterocyclic group containing from 1 to 3 heteroatoms selected from O, S and N, wherein the ortho, meta or para is more Independently selected from one of the following substituents: hydrogen, (CH ₂ ) _n alkyl, (CH ₂ ) _n hydroxy, (CH ₂ ) _n C _3-12 cycloalkyl, (CH ₂ ) _n C _{3- 12} nitrogen-containing cycloalkyl, (CH ₂ ) _n benzene ring, aldehyde group and (CH ₂ ) _n COC _3-12 nitrogen-containing cycloalkyl group; wherein n 10;v)NH(CH ₂ ) _n R ₁ ,0 n 10, R ₁ is selected from one of the following substituents: N(CH ₃ ) ₂ , C(NH ₂ ) ₂ , a linear alkyl group of NH(CH ₂ ) _n H , an alkyl group having a branched chain, An alkyl group having an amino group substitution, an alkyl group having a hydroxyl group substitution; vi) NH(CH ₂ ) _n R ₂ , 0 n 10, R ₂ is selected from one of the following substituents: a benzene ring, a C _3-12 cycloalkyl group and a heterocyclic group, containing from 1 to 3 hetero atoms selected from O, S and N, wherein the The position, the meta position or the para position may be independently selected from one of the following substituents: a methoxy group, an amine group, a benzene ring, an alkyl group having a C1-C3 branch substitution, an amine group, a nitro group, a hydroxyl group, and a C _3-12 heterocyclic ring; wherein the C _3-12 heterocyclic ring contains from 1 to 3 heteroatoms selected from O, S and N; and pharmaceutically acceptable salts, stereoisomers and mirror image isomers .

To achieve the aforementioned object, the substituent group i)-vi) is selected from the group consisting of chlorine, hydroxyl, methoxy, 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, morpholine , thiomorpholino, piperidin-1-yl, 4-hydroxypiperidin-1-yl, 4-benzylpiperidin-1-yl, [1,4'-bipiperidinyl]-1'- , 4-(3-(piperidin-4-yl)propyl, pyrrolidin-1-yl, 2-oxopiperidin-1-yl, methylamino, ethylamino, propylamino, butyl Amino, isobutyl, pent-3-ylamino, (2-(dimethylamino)ethyl)amino, (2-(diethylamino)ethyl)amino, 2-ethanolamino, 3-propanol Amino, 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-indol-2-yl)amino, cyclohexyl, (1-benzylpiperidin-4-yl) Amino, (thiophen-2-yl) 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-aminophenylethyl) a group consisting of amino, anthracene and piperidin-1-ylamino.

To achieve the foregoing objects, the compound is selected from the group consisting of compounds 1-21 and N1-N34.

3-((4-chlorophenyl)thio)-2-hydroxyquinoline-4-carboxylic acid, 6,9-dichloro-12H-dehydrothiamine [2,3-c]quinoline-12- Ketone, 10-chloro-6-hydroxy-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-methoxy-12H-dehydrothiamine and [2 ,3-c]quinolin-12-one, 10-chloro-6-dimethylamino-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-( Piperazin-1-yl)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(4-methylpiperazin-1-yl)-12H- Dehydrothiamine-[2,3-c]quinolin-12-one, 10-chloro-6-(4-ethylpiperazin-1-yl)-12H-dehydrothiamine[2,3- c] quinoline-12-one, 10-chloro-6-(4-(2-hydroxyethyl)piperazin-1-yl)-12H-dehydrothieno[2,3-c]quinoline 12 -ketone, 6-(4-benzylpiperazin-1-yl)-10-chloro-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(4-phenylpiperazin-1-yl)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-morpholino- 12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-thiomorpholino-12H-dehydrothieno[2,3-c]quinoline- 12-keto, 10-chloro-6-(piperidin-1-yl)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(4-hydroxyl Piperidin-1-yl)-12H-dehydrothieno[2,3-c]quinolin-12-one, 6-(4-benzylpiperidin-1-yl)-10-chloro-12H- Dehydrothiamine-[2,3-c]quinolin-12-one, 6-([1,4'-bipiperidinyl]-1'-yl)-10-chloro-12H-dehydrothiamine [2,3-c]quinolin-12-one, 10-chloro-6-(4-(3-(piperidin-4-yl)propyl)piperidin-1-yl-12H-dehydrothiamine And [2,3-c]quinolin-12-one, 10-chloro-6-(pyrrolidin-1-yl)-12H-dehydrothieno[2,3-c]quinolin-12-one , 10-chloro-6-(2-oxopiperidin-1-yl)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-methylamino -12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-ethylamino-12H-dehydrothieno[2,3-c]quinoline-12 -ketone, 10-chloro-6-propylamino-12H-dehydrothieno[2,3-c]quinolin-12-one, 6-(butylamino)-10-chloro-12H-dehydrogenation Thiamin[2,3-c]quinolin-12-one, 10-chloro-6-isobutyl -12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(pent-3-ylamino)-12H-dehydrothiamine[2,3- c] quinoline-12-one, 10-chloro-6-((2-(dimethylamino)ethyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12- Ketone, 10-chloro-6-((2-(diethylamino)ethyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6 -(2-ethanolamino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(3-propanolamino)-12H-dehydrothiamine [2,3-c]quinolin-12-one, 10-chloro-6-(5-pentanolamino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10 -Chloro-6-((1-hydroxybutan-2-yl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-((4) -methylpentan-2-yl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 6-((2-aminoethyl)amino)-10-chloro- 12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-((2-((2-hydroxyethyl))amino)ethyl)amino)-12H-de Hydrogenthio[2,3-c]quinolin-12-one, 10-chloro-6-((2-morpholinoethyl)amino)-12H-dehydrothiamine[2,3-c Quinoline-12-one, 10-chloro-6-((3-(dimethylamino)propyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one , 10-chloro-6-((3- (diethylamino)propyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-((3-((2-hydroxyethyl))amino)propyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro -6-((2,3-dihydro-1H-indol-2-yl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6- (cyclohexyl)-12H-dehydrothieno[2,3-c]quinolin-12-one, 6-((1-benzylpiperidin-4-yl)amino)-10-chloro-12H- Dehydrothiamine[2,3-c]quinolin-12-one, 10-chloro-6-((thiophen-2-ylmethyl)amino)-12H-dehydrothiamine[2,3- c] quinoline-12-one, 10-chloro-6-((cyclohexylmethyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 6-(benzyl Base amino)-10-chloro-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-((pyridin-2-ylmethyl)amino)-12H- Dehydrothiamine-[2,3-c]quinolin-12-one, 6-((benzo[d][1,3]dioxol-5-ylmethyl)amino)-10 -Chloro-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-((2-methoxybenzyl)amino)-12H-dehydrothiamine and [ 2,3-c]quinolin-12-one, 10-chloro-6-((3,4-dimethoxybenzyl)amino)-12H-dehydrothieno[2,3-c]quina Porphyrin-12-one, 10-chloro-6-(phenethylamino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(4- Methoxyphenethyl)amino)-1 2H-dehydrothieno[2,3-c]quinolin-12-one, 6-((4-aminophenethyl)amino)-10-chloro-12H-dehydrothiamine [2,3 -c]quinolin-12-one, 2-(10-chloro-12-oxo-12H-dehydrothieno[2,3-c]quinolin-6-yl)anthracene, 10-chloro-6 -(piperidin-1-ylamino)-12H-dehydrothieno[2,3-c]quinolin-12-one, and salts thereof.

Another object of the present invention is to provide a pharmaceutical composition comprising a therapeutically effective amount of a compound as described above, and at least one pharmaceutically acceptable carrier, diluent or excipient.

Another object of the present invention is to provide a use of a compound as described above for the manufacture of a medicament useful for inhibiting the activity of a first type of topoisomerase.

Another object of the present invention is to provide a use of a compound as described above for the manufacture of a medicament useful for inhibiting the activity of a second type of topoisomerase.

Another object of the present invention is to provide a use of a compound as described above for the manufacture of a medicament useful for the treatment of cancer.

To achieve the aforementioned object of the invention, wherein the cancer is selected from leukemia, non-small cells Lung cancer, colorectal cancer, central nervous cancer, melanoma, ovarian cancer, kidney cancer, prostate cancer and breast cancer.

Another object of the present invention is to provide a process for producing a compound according to claim 1, which comprises: (1) isatin, 2-((chlorophenyl)thio)acetic acid and sodium acetate The mixture was reacted at 150 ° C for 1 hour, and the reaction mixture was cooled, and then acetic acid was added to obtain a precipitate, which was washed with acetic acid, water and n-hexane to obtain compound 2 (3-((4-chlorophenyl)thio). 2-hydroxyquinoline-4-carboxylic acid); (2) Compound 2 (3-((4-chlorophenyl)thio)-2-hydroxyquinoline-4-carboxylic acid) is dissolved in trichloroox Phosphorus at 150 ° C for 48 hours. After cooling, the reaction was poured into water. The precipitate was collected by filtration and placed in a 10% sodium hydrogen carbonate solution and stirred for 1 hour. The precipitate was collected and rinsed with water. The crude product was recrystallized from dichloromethane. Compound 3 (6,9-dichloro-12H-dehydrothiamine [2,3-c]quinolin-12-one); (3) Compound 3 (6,9-dichloro-12H-dehydrosulfide) The amine [2,3-c]quinolin-12-one) was dissolved in DMF, concentrated hydrochloric acid was added and refluxed for six hours, then concentrated hydrochloric acid was added dropwise and refluxed for further 12 hours, the reaction was dehydrated in vacuo, then water was added and filtered. The crude product is washed with ethanol to give compound 4 (10-chloro-6- Hydroxy-12H-dehydrothieno[2,3-c]quinolin-12-one); (4) Suspended compound 3 in methanol (6,9-dichloro-12H-dehydrothiamine [2 , 3-c]quinoline-12-one) was refluxed with sodium methoxide for 16 hours. After cooling, the solvent was removed, filtered and washed with ethanol and n-hexane to give compound 5 (10-chloro-6-methoxy-12H -dehydrothieno[2,3-c]quinolin-12-one); (5) Compound 3 (6,9-dichloro-12H-dehydrothiamine [2,3-c]quinoline -12-ketone) and a suitable secondary amine and sodium carbonate dissolved in DMSO or DMF, reflux for 10 hours, after standing and cooling, the reactants were added to ice water, and the precipitate was filtered, washed with water and methanol to obtain a compound. 6-21: 10-Chloro-6-dimethylamino-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(piperazin-1-yl)- 12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(4-methylpiperazin-1-yl)-12H-dehydrothiamine [2, 3-c]quinolin-12-one, 10-chloro-6-(4-ethylpiperazin-1-yl)-12H-dehydrothieno[2,3-c]quinolin-12-one , 10-chloro-6-(4-(2-hydroxyethyl)piperazin-1-yl)-12H-dehydrothieno[2,3-c]quinolin-12-one, 6-(4-Benzylpiperazin-1-yl)-10-chloro-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(4-benzene Piperazin-1-yl)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-morpholino-12H-dehydrothiamine [2, 3-c]quinolin-12-one, 10-chloro-6-thiomorpholino-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6- (piperidin-1-yl)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(4-hydroxypiperidin-1-yl)-12H- Dehydrothiamine-[2,3-c]quinolin-12-one, 6-(4-benzylpiperidin-1-yl)-10-chloro-12H-dehydrothiamine[2,3- c] quinoline-12-one, 6-([1,4'-bipiperidinyl]-1'-yl)-10-chloro-12H-dehydrothieno[2,3-c]quinoline- 12-keto, 10-chloro-6-(4-(3-(piperidin-4-yl)propyl)piperidin-1-yl-12H-dehydrothieno[2,3-c]quinoline -12-ketone, 10-chloro-6-(pyrrolidin-1-yl)-12H-dehydrothieno[2,3-c]quinolin-12-one, and 10-chloro-6-(2 -oxopiperidin-1-yl)-12H-dehydrothieno[2,3-c]quinolin-12-one; (6) Compound 3 (6,9-dichloro-12H-dehydrogenation) Thiamine [2,3-c]quinolin-12-one) was dissolved in DMSO with an appropriate primary amine and refluxed for 8 hours. After cooling, the reaction was taken in 100 mL water.

After the precipitate was filtered off, it was washed with water and methanol to give Compound N1-N34: 10-chloro-6-methylamino-12H-dehydrothiamine[2,3-c]quinolin-12-one, 10-chloro-6-ethylamino-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-propylamino-12H-dehydrothiamine and [2 ,3-c]quinolin-12-one, 6-(butylamino)-10-chloro-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6 -isobutyl-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(pent-3-ylamino)-12H-dehydrothiamine and [2 ,3-c]quinolin-12-one, 10-chloro-6-((2-(dimethylamino)ethyl)amino)-12H-dehydrothieno[2,3-c]quinoline -12-ketone, 10-chloro-6-((2-(diethylamino)ethyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10- Chloro-6-(2-ethanolamino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(3-propanolamino)-12H-dehydrogenation Thiamin[2,3-c]quinolin-12-one, 10-chloro-6-(5-pentanolamino)-12H-dehydrothieno[2,3-c]quinolin-12- Ketone, 10-chloro-6-((1-hydroxybutan-2-yl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6- ((4-methylpentan-2-yl)amino)-12H-dehydrothieno[2,3-c]quinoline- 12-keto, 6-((2-aminoethyl)amino)-10-chloro-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(( 2-((2-Hydroxyethyl)amino)ethyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-((2-? Oleinoethyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-((3-(dimethylamino)propyl)amino) -12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-((3-(diethylamino)propyl)amino)-12H-dehydrothiamine [2,3-c]quinolin-12-one, 10-chloro-6-((3-((2-hydroxyethyl))amino)propyl)amino)-12H-dehydrothiamine [2, 3-c]quinolin-12-one, 10-chloro-6-((2,3-dihydro-1H-indol-2-yl)amino)-12H-dehydrothiamine[2,3-c Quinoline-12-one, 10-chloro-6-(cyclohexyl)-12H-dehydrothieno[2,3-c]quinolin-12-one, 6-((1-benzylpiperidine) 4-yl)amino)-10-chloro-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-((thiophen-2-ylmethyl)amino )-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-((cyclohexylmethyl)amino)-12H-dehydrothiamine [2,3 -c]quinoline-12-one, 6-(benzylamino)-10-chloro-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-( (pyridin-2-ylmethyl) Amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 6-((benzo[d][1,3]dioxol-5-ylmethyl Amino)-10-chloro-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-((2-methoxybenzyl)amino)-12H-de Hydrogenthio[2,3-c]quinolin-12-one, 10-chloro-6-((3,4-dimethoxybenzyl)amino)-12H-dehydrothiamine [2, 3-c]quinoline-12-one, 10-chloro-6-(phenethylamino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6 -((4-methoxyphenethyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 6-((4-aminophenethyl)amino)- 10-chloro-12H-dehydrothieno[2,3-c]quinolin-12-one, 2-(10-chloro-12-oxo-12H-dehydrothiamine[2,3-c Quinoline-6-yl)indole, and 10-chloro-6-(piperidin-1-ylamino)-12H-dehydrothieno[2,3- c ]quinolin-12-one.

Figure 1 is a summary of the reaction of chemical synthesis of dehydrothiamine [2,3- c ]quinolin-12-one derivatives.

Figure 2 shows the effect of compounds 5, 7, 8, 16, 19 and CPT (camptothecin) on 25, 50 μM for the unwinding of DNA acting on the first type of topoisomerase (TOP I).

Figure 3 shows the effect of compounds N2, N7, N8, N9, N14-N19, N25 and CPT at 50 μM on the unwinding of DNA acting on the first type of topoisomerase (TOP I).

Figures 4a-d show the effect of Compound 7, N7, N14, N15, N18, N19, N25 and CPT at 1-50 μM for the unwinding of DNA acting on the first type of topoisomerase (TOP I).

Figure 5 shows compounds 5, 7, 8, 16, 19 and VP-16 (etoposide, as a positive control) at 25, 50 μM for the action of the second type of topoisomerase (TOP II) DNA. The effect of unwinding.

Figure 6 shows the effect of the compounds N2, N7, N8, N9, N14-N19, N25 and VP-16 at 50 μM on the unwinding of DNA acting on the first type of topoisomerase (TOP I).

Figure 7a-d shows the effect of compound 7, N7, N8, N14, N15, N18, N19, N25 and VP-16 at 1-50 μM for DNA unwinding of the first type of topoisomerase (TOP I) .

All of the technical and scientific terms described in this specification, unless otherwise defined, are intended to be common to those of ordinary skill in the art.

All of the technical and scientific terms described in this specification, unless otherwise defined, are intended to be common to those of ordinary skill in the art.

The terms "treatment," "in therapy," and the like, are meant to delay, ameliorate, reduce, or reverse a condition that is currently afflicting the condition of the patient or any condition associated with the condition and preventing the condition or any symptoms thereof. Methods.

The term "pharmaceutically acceptable" means that the substance or composition must be compatible with the other ingredients of the formulation and not deleterious to the patient.

The composition of the present invention can be applied by techniques well known to those skilled in the art. The Lactobacillus isolate, and a pharmaceutically acceptable vehicle, are prepared in a dosage form suitable for use in the compositions of the present invention. Wherein the dosage form comprises, but is not limited to, a solution, an emulsion, a suspension, a powder, a tablet, a pill, a lozenge, a tablet. (troche), chewing gum, slurries, and other dosage forms similar or suitable for use in the present invention.

Wherein the pharmaceutically acceptable carrier may comprise one or more agents selected from the group consisting of: solvents, emulsifiers, suspending agents, decomposers, binding agents ), excipient, stabilizing agent, chelating agent, diluent, gelling agent, preservative, lubricant, surface Active agent, and other carriers similar or suitable for use in the present invention.

In the above composition, one or more dissolution aids, buffers, preservatives, coloring agents, perfumes, flavoring agents and the like which are usually used in the field of preparation may be appropriately added as needed.

The term "pharmaceutically acceptable excipient" includes, but is not limited to, polymers, resins, plasticizers, fillers, lubricants, diluents, binders, disintegrants, solvents, co-solvents, surfactants At least one of a preservative, a sweetener, a flavoring agent, a pharmaceutical grade dye or pigment, and a viscosity agent.

The term "pharmaceutical composition" is a solid or liquid composition in a form, concentration and purity suitable for administration to a patient (eg, a human or animal patient) which, after administration, induces a desired Physiological changes. Pharmaceutical compositions are typically sterile and/or non-pyrogenic.

The present invention is exemplified by the following examples, but the present invention is not limited by the following examples. The drugs and biological materials used in the present invention are commercially available and are readily available. The following are only examples of available pipelines.

All reactions were monitored by thin layer chromatography (TLC) coated with silicone 60 F ₂₅₄ . The melting points of all synthetic compounds were determined by a Büchi B-545 assay device. ¹ H NMR: Varian GEMINI-300 (300 MHz) or Agilent 400 MR DD2 (400 MHz); δ values are in ppm relative to tetramethyl decane (TMS) as internal standard (0 ppm). Multiplicities are denoted 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). High resolution electrospray ionization (HRESI): Finnigan MAT 95S (Instrument Center, National Taiwan University). X-ray single crystal diffraction: Bruker Enraf-Nonius APEX II diffractometer (Department of Chemistry, National Taiwan Normal University). A typical procedure for the preparation of thiochromenoquinolones is indicated in the typical experiment and is described below (Figure 1).

General rules for chemical synthesis

Synthesis Step General A : Synthesis of Compound 2.

Isatin (0.44g, 2.99mmole) and 2-((4-chlorophenyl)thio)acetic acid (0.70g, 3.47mmol) and sodium acetate (0.05g) were mixed and reacted in miniclave (150 °C) for 1 hour (thin- Layer chromatography, TLC monitoring). After the reaction mixture was cooled, acetic acid (10 mL) was added to obtain a gray precipitate, which was washed with acetic acid, water and n-hexane to give a bright purple compound.

Synthesis Step General B : Synthesis of Compound 3.

Compound 2 was dissolved in phosphorus oxychloride (5 mL) at 150 ° C for 48 hours. After cooling, the reaction mixture was poured into ice water at 0 ° C. The green precipitate was collected by filtration and placed in a 10% sodium hydrogen carbonate solution (50 mL) and stirred vigorously for 1 hour. The precipitate was collected and rinsed with water. The crude product was recrystallized from dichloromethane to give the title compound.

Synthesis Procedure General C : Synthesis of Compound 4.

Compound 3 (0.32 g, 0.96 mmol) was dissolved in DMF (20 mL). After six hours, concentrated hydrochloric acid (6 mL) was added dropwise and refluxed for a further 12 hr. The reactant was dehydrated in vacuo, and water (20 mL) was added. After filtration, the crude product was washed with ethanol to give Compound 4 as a yellow solid.

Synthesis Step General D : Synthesis of Compound 5.

Suspension Compound 3 (0.33 g, 1.0 mmol) in 20 mL of MeOH was refluxed with sodium methoxide (0.55 g, 10 mmol). After cooling, the solvent was removed by a rotary evaporation vacuum apparatus, filtered, and washed with ethanol and n-hexane to afford compound 5 as a white solid.

Synthesis Procedure General E : Synthesis of Compound 6-21.

Take compound 3 (0.33 g, 1.0 mmol) with an appropriate secondary amine (1.1 mmol) Sodium carbonate (5 mmol) was dissolved in DMSO or DMF (20 mL) and refluxed for 10 h (TLC). After standing to cool, the reactant was added to 100 mL of ice water. The precipitate was filtered off and washed with water and methanol to give a yellow solid.

Synthesis Procedure General F : Synthesis of Compound N1-34.

Compound 3 (0.33 g, 1.0 mmol) was taken in EtOAc (30 mL) elute After cooling, the reaction was added to 100 mL of water. The precipitate was filtered off and washed with water and methanol to give a yellow solid.

Example 1

3-((4-chlorophenyl)thio)-2-hydroxyquinoline-4-carboxylic acid (2)

3-((4-Chlorophenyl)thio)-2-hydroxyquinoline-4-carboxylic acid(TC-SCl)(2)

The pure compound was 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[MH] ^- 330.0002.

Example 2

6,9-Dichloro-12H-dehydrothiamine [2,3-c]quinolin-12-one (3)

6,9-Dichloro-12H-thiochromeno[2,3-c]quinolin-12-one(3)

A yellow solid was isolated (yield 90%). ( 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 (100MHz, 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-dehydrothieno[2,3-c]quinolin-12-one (4)

10-Chloro-6-hydroxy-12H-thiochromeno[2,3-c]quinolin-12-one(4)

A yellow solid was isolated (yield 95%). ( R _f =0.40 at EA).Mp>410 ° C. ¹ H NMR (400 MHz, DMSO- d 6): 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-dehydrothieno[2,3-c]quinolin-12-one (5)

10-Chloro-6-methoxy-12H-thiochromeno[2,3-c]quinolin-12-one(5)

A gray solid was isolated (yield 91%). ( 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 (100MHz, 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-dehydrothieno[2,3-c]quinolin-12-one (6)

10-Chloro-6-dimethylamino-12H-thiochromeno[2,3-c]quinolin-12-one(6)

A yellow solid was isolated (yield 85%). ( 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, 127.78128.47,129.07,129.37,130.59,130.67,132.23,132.46,133.61,134.36,144.84,158.32,181.52( 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-dehydrothieno[2,3-c]quinolin-12-one (7)

10-Chloro-6-(piperazin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one(7)

A yellow solid was isolated (yield 69%). ( 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 (100MHz, 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-dehydrothieno[2,3-c]quinolin-12-one (8)

10-Chloro-6-(4-methylpiperazin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one(8)

A yellow-green solid was isolated (yield 80%). ( 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-dehydrothieno[2,3-c]quinolin-12-one (9)

10-Chloro-6-(4-ethylpiperazin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one(9)

A yellow solid was isolated (yield 74%). ( 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-dehydrothieno[2,3-c]quinolin-12-one (10)

10-Chloro-6-(4-(2-hydroxyethyl)piperazin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12one(10)

A yellow-green solid was isolated (yield 60%). ( 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-dehydrothieno[2,3-c]quinolin-12-one (11)

6-(4-Benzylpiperazin-1-yl)-10-chloro-12H-thiochromeno[2,3-c]quinolin-12-one(11)

A yellow solid was isolated (yield 81%). ( 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-dehydrothieno[2,3-c]quinolin-12-one (12)

10-Chloro-6-(4-phenylpiperazin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one(12)

A yellow solid was isolated (yield 77%). ( 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 (75MHz, 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-dehydrothieno[2,3-c]quinolin-12-one (13)

10-Chloro-6-mo _r pholino-12H-thiochromeno[2,3-c]quinolin-12-one(13)

A yellow solid was isolated (yield 70%). ( 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-dehydrothieno[2,3-c]quinolin-12-one (14)

10-Chloro-6-thiomorpholino-12H-thiochromeno[2,3-c]quinolin-12-one(14)

A yellow solid was isolated (yield 86%). ( 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 (75MHz, 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-dehydrothieno[2,3-c]quinolin-12-one (15)

10-Chloro-6-(piperidin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one(15)

A yellow solid was isolated (yield 86%). ( 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-dehydrothieno[2,3-c]quinolin-12-one (16)

10-Chloro-6-(4-hydroxypiperidin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one(16)

A yellow-gray solid was isolated (yield 84%). ( 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, piperididine-CH _a ), 2.15-2.21 (2H, m, piperididine-CH _e ), 3.19 (2H, td, J = 8.4, 2.1 Hz,piperidine-NCH _a ), 3.60-3.65 (2H, m, piperididine-NCH _e ), 4.01 (1H, sext, J = 3.0 Hz, piperididine-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 = 63, 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; M+H] ⁺ 397.0757.

Example 16

6-(4-Benzylpiperidin-1-yl)-10-chloro-12H-dehydrothieno[2,3-c]quinolin-12-one (17)

6-(4-Benzylpiperidin-1-yl)-10-chloro-12H-thiochromeno[2,3-c]quinolin-12-one(17)

A yellow solid was isolated (yield 90%). ( 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, piperididine-CH ₂ ), 2.71 (2H, d, J = 5.1 Hz, _{piperidine-CH 2), 3.00 (} 2H, td, J = 9.3,1.2Hz, -NCH 2 -), 3.65 (2H, d, J = 9.3Hz, -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'-bipiperidinyl]-1'-yl)-10-chloro-12H-dehydrothieno[2,3-c]quinolin-12-one (18)

6-([1,4'-Bipiperidin]-1'-yl)-10-chloro-12H-thiochromeno[2,3-c]quinolin-12-one(18)

A yellow solid was isolated (yield 92%). ( 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, piperididine-H), 1.86-1.98 (2H, qd, J = 12.4, 2.8 Hz, piperididine-H), 2.06 (2H, d, J = 11.6 Hz, piperididine-H), 2.54 (1H) , t, J = 10.8 Hz, piperidine-H), 2.65 (3H, br, piperididine-H), 3.05 (2H, t, J = 12 Hz, piperididine-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] ⁺ 4464.1593.

Example 18

10-chloro-6-(4-(3-(piperidin-4-yl)propyl)piperidin-1-yl-12H-dehydrothieno[2,3-c]quinolin-12-one (19)

10-Chloro-6-(4-(3-(piperidin-4-yl)propyl)piperidin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one(19)

A yellow solid was isolated (yield 76%). ( 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-dehydrothieno[2,3-c]quinolin-12-one (20)

10-Chloro-6-(pyrrolidin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one(20)

A yellow solid was isolated (yield 86%). ( 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-dehydrothieno[2,3-c]quinolin-12-one (21)

10-Chloro-6-(2-oxopiperidin-1-yl)-12H-thiochromeno[2,3-c]quinolin-12-one(21)

Compound 3 (0.33 g, 1.0 mmol) and piperidin-2-one (5.55 mmol) and sodium carbonate (5 mmol) were dissolved in DMF (15 mL) and refluxed for 10 hours (TLC). After standing for 10 minutes to cool, the reaction was added to 100 mL of ice water. The precipitate was filtered off and washed with water and methanol to give a yellow solid. A yellow solid was isolated (yield 89%). 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-dehydrothieno[2,3-c]quinolin-12-one (N1)

10-Chloro-6-methylamino-12H-thiochromeno[2,3-c]quinolin-12-one(N1)

A yellow solid was isolated (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-dehydrothieno[2,3-c]quinolin-12-one (N2)

10-Chloro-6-ethylamino-12H-thiochromeno[2,3-c]quinolin-12-one(N2)

A yellow solid was isolated (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-dehydrothieno[2,3-c]quinolin-12-one (N3)

10-Chloro-6-propylamino-12H-thiochromeno[2,3-c]quinolin-12-one(N3)

A yellow solid was isolated (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-dehydrothieno[2,3-c]quinolin-12-one (N4)

6-(Butylamino)-10-chloro-12H-thiochromeno[2,3-c]quinolin-12-one(N4)

A yellow solid was isolated (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-isobutyl-12H-dehydrothieno[2,3-c]quinolin-12-one (N5)

10-Chloro-6-isobutylamino-12H-thiochromeno[2,3-c]quinolin-12-one(N5)

A yellow solid was isolated (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-(pent-3-ylamino)-12H-dehydrothieno[2,3-c]quinolin-12-one (N6)

10-Chloro-6-(pentan-3-ylamino)-12H-thiochromeno[2,3-c]quinolin-12-one(N6)

A yellow solid was isolated (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,[MH] ^- :381.0851.

Example 27

10-chloro-6-((2-(dimethylamino)ethyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one (N7)

10-Chloro-6-((2-(dimethylamino)ethyl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one(N7)

A yellow solid was isolated (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-dehydrothieno[2,3-c]quinolin-12-one (N8)

10-Chloro-6-((2-(diethylamino)ethyl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one(N8)

A yellow solid was isolated (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 (100MHz, 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-dehydrothieno[2,3-c]quinolin-12-one (N9)

10-Chloro-6-(2-ethanolamino)-12H-thiochromeno[2,3-c]quinolin-12-one(N9)

A yellow solid was isolated (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-dehydrothieno[2,3-c]quinolin-12-one (N10)

10-Chloro-6-(3-propanolamino)-12H-thiochromeno[2,3-c]quinolin-12-one(N10)

A yellow solid was isolated (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-dehydrothieno[2,3-c]quinolin-12-one (N11)

10-Chloro-6-(5-pentanolamino)-12H-thiochromeno[2,3-c]quinolin-12-one(N11)

A yellow solid was isolated (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-dehydrothieno[2,3-c]quinolin-12-one (N12)

10-Chloro-6-((1-hydroxybutan-2-yl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one(N12)

A yellow solid was isolated (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-dehydrothieno[2,3-c]quinolin-12-one (N13)

10-Chloro-6-((4-methylpentan-2-yl)amino)-12H-thiochromeno [2,3-c]quinolin-12-one(N13)

A yellow solid was isolated (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-dehydrothieno[2,3-c]quinolin-12-one (N14)

6-((2-Aminoethyl)amino)-10-chloro-12H-thiochromeno[2,3-c]quinolin-12-one(N14)

A yellow solid was isolated (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-dehydrothiamine [2,3-c]quinolin-12-one (N15)

10-Chloro-6-((2-((2-hydroxyethyl)amino)ethyl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one(N15)

A yellow solid was isolated (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-dehydrothieno[2,3-c]quinolin-12-one (N16)

10-Chloro-6-((2-morpholinoethyl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one(N16)

A yellow solid was isolated (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,[MH] ^- :424.0885.

Example 37

10-chloro-6-((3-(dimethylamino)propyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one (N17)

10-Chloro-6-((3-(dimethylamino)propyl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one(N17)

A yellow solid was isolated (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. , 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-dehydrothieno[2,3-c]quinolin-12-one (N18)

10-Chloro-6-((3-(diethylamino)propyl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one(N18)

A yellow solid was isolated (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,[MH] ^- :424.1284.

Example 39

10-chloro-6-((3-((2-hydroxyethyl))amino)propyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one (N19)

10-Chloro-6-((3-((2-hydroxyethyl)amino)propyl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one(N19)

A tan solid was isolated (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 & -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 = 8.0Hz, Ar-H), 8.46 (1H, d, J = 2.0Hz, Ar-H), 9.39 (1H, d, J = 8.4Hz, Ar-H). 13 C NMR (100MHz, 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-indol-2-yl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one (N20)

10-Chloro-6-((2,3-dihydro-1H-inden-2-yl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one(N20)

A tan solid was isolated (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-(cyclohexyl)-12H-dehydrothieno[2,3-c]quinolin-12-one (N21)

10-Chloro-6-(cyclohexylamino)-12H-thiochromeno[2,3-c]quinolin-12-one(N21)

A tan solid was isolated (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-dehydrothieno[2,3-c]quinolin-12-one (N22)

6-((1-Benzylpiperidin-4-yl)amino)-10-chloro-12H-thiochromeno[2,3-c]quinolin-12-one(N22)

A tan solid was isolated (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, piperididine-H), 2.92 (2H, d, J = 11.6 Hz, piperididine-H), 3.59 (2H, s, - CH ₂ -), 4.35 (1H, sext, J = 6.4 Hz, piperididine-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-dehydrothieno[2,3-c]quinolin-12-one (N23)

10-Chloro-6-((thiophen-2-ylmethyl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one(N23)

A tan solid was isolated (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,[MH] ^- :407.0085.

Example 44

10-chloro-6-((cyclohexylmethyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one (N24)

10-Chloro-6-((cyclohexylmethyl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one(N24)

A tan solid was isolated (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-dehydrothieno[2,3-c]quinolin-12-one (N25)

6-(Benzylamino)-10-chloro-12H-thiochromeno[2,3-c]quinolin-12-one(N25)

A tan solid was isolated (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-dehydrothieno[2,3-c]quinolin-12-one (N26)

10-Chloro-6-((pyridin-2-ylmethyl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one(N26)

A tan solid was isolated (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 & 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-dehydrothieno[2,3-c]quinoline -12-ketone (N27)

6-((Benzo[d][1,3]dioxol-5-ylmethyl)amino)-10-chloro-12H-thiochromeno[2,3-c]quinolin-12-one(N27)

A tan solid was isolated (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, [MH] ^- : 445.0440.

Example 48

10-chloro-6-((2-methoxybenzyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one (N28)

10-Chloro-6-((2-methoxybenzyl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one(N28)

A tan solid was isolated (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-dehydrothieno[2,3-c]quinolin-12-one (N29)

10-Chloro-6-((3,4-dimethoxybenzyl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one(N29)

A tan solid was isolated (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) m/z Calcd for C ₂₅ H ₁₉ N ₂ O ₃ SCl[M] ⁺ : 462.0805;found[M+H] ⁺ :463.0900,[MH] ^- :461.0754.

Example 50

10-chloro-6-(phenethylamino)-12H-dehydrothieno[2,3-c]quinolin-12-one (N30)

10-Chloro-6-(phenethylamino)-12H-thiochromeno[2,3-c]quinolin-12-one(N30)

A tan solid was isolated (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-dehydrothieno[2,3-c]quinolin-12-one (N31)

10-Chloro-6-((4-methoxyphenethyl)amino)-12H-thiochromeno[2,3-c]quinolin-12-one(N31)

A yellow solid was isolated (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-dehydrothieno[2,3-c]quinolin-12-one (N32)

6-((4-Aminophenethyl)amino)-10-chloro-12H-thiochromeno[2,3-c]quinolin-12-one(N32)

A yellow solid was isolated (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-12 H -dehydrothieno[2,3- c ]quinolin-6-yl)indole

2-(10-Chloro-12-oxo-12 H- thiochromeno[2,3- c ]quinolin-6-yl)g uanidine(N33)

A yellow solid was isolated (yield 85%). Mp: 370 ° C. ¹ H NMR (400 MHz, DMSO- d ₆ ): δ ppm. 7.40 (3H, td, J = 8.4, 1.2 Hz, Ar-H & -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 ( 100MHz, 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)-12 H -dehydrothieno[2,3- c ]quinolin-12-one (N34)

10-Chloro-6-(piperidin-1-ylamino)-12 H- thiochromeno[2,3- c ]quinolin-12-one(N34)

A yellow solid was isolated (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, 181.88.

Pharmacological activity experiment

In the pharmacological test, the chemically synthesized compound structures 2-21 and N -1 to N-34 (a total of 54 drugs) were tested for the following five parts: 1. Cell survival assay (MTT assay) Test; Second, type I and type II topoisomerase activity test; third, cell cycle arrest; fourth, apoptosis related protein expression; five, the United States Cancer Research Center (NCI) screened out 26 compounds Structure, 60 cancer cell strains were tested for the 26 compounds.

Example 55 Cell Survival Assay (MTT assay) assay

All synthesis of compounds using the MTT assay, cytotoxic compounds were analyzed PC-3 (BCRC 60122, Taiwan ) and ^{DU-145 (HTB-81 TM} , ATCC, Rockville, MD) cell lines. The above cells were cultured in RPMI-1640, 5% FBS (v/v), 100 U/mL penicillin and 50 mg/mL streptomycin. Approximately 2 x 10 ³ cells were cultured in a 96-well plate at 5% CO ₂ for 24 hours at 37 °C. To measure in vitro cytotoxicity, all compounds were dissolved in DMSO prior to the experiment and pre-mixed in the same cell culture medium for administration and adjusted for different dosing concentrations (0.15, 0.5, 1.5, 5, 15 μM). ) Perform three replicate experiments. At 72 hours after administration, 100 μL of MTT (1 mg/mL) was added to each well and cultured at 37 ° C for 4 hours. After removing the MTT solution, 100 μL of DMSO was added to each well, followed by incubation at 37 ° C for 20 minutes. The absorbance spectrum was measured at 560 nm using an ELISA reader. Results presented at least three replicates, values in Table 1 presents the IC _50.

Further, the compounds N7, N8, N14, N15, N17 and N18 contain at least one nitrogen atom in the side branch, which shows superior cytotoxic activity compared to only a single hydroxyl group, an alkyl group or an aromatic ring. Compounds 5, 7, 8, 16, 19, N2, N7, N8, N9, N14, N15, N16, N17, N18, N19 and N25 were screened for topoisomerase activity assays.

Example 56 First and second type topoisomerase activity test

Compounds 5, 7, 8, 16, 19, N2, N7, N8, N9, N14, N15, N16, N17, N18, N19 and N25 were tested at 25 μM or 50 μM for topoisomerase activity (Figure 2-4 ).

In the first type of topoisomerase activity test, compounds 7, N7, N14, N15, N17, N18 and N25 were more inhibitory than camptothecin, and further experiments were carried out at five different concentrations (Fig. 4a-d). The IC ₅₀ values of Compound 7, N7, N14, N15, N17, N18 and N25 were 10, 10, 1, 5, 25, 5 and 25 μM, respectively (measured using TopoGEN TG2005H, TG-2000H-1).

In the second type of topoisomerase activity test (Fig. 5-7), compounds 7, N7, N8, N14, N15, N18 and N19 have more inhibitory effects than camptothecin, and further five Experiments were performed at different concentrations (Fig. 7a-d). The IC ₅₀ values of Compound 7, N7, N8, N14, N15, N18 and N19 were 10, 10, 1, 10, 5, 1 and 1 μM, respectively (measured using TopoGEN TG2005H, TG-2000H-1).

Example 57 Screening by the American Cancer Research Center (NCI)

The American Cancer Research Center (NCI) screened 26 compound structures (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), a poisoning test of 60 cancer cell lines against these 26 compounds. In the first stage, 26 compounds were first subjected to a 10 μM poisoning test, and a poisoning test of 60 cancer cell lines was performed. After 48 hours, a cell survival test (Sulforhodamine B, SRB) was performed. The results are shown in Table 2 - Table 4, expressed as growth percent.

Then, five compounds with more potential for toxic killing were found out: N2, N7, N14, N19 and N25 were tested for the second phase (0.01, 0.1 , 1, 10, 100 μM) (Table 5).

The above-mentioned multiple functions are in fact the statutory invention patents that fully meet the novelty and progressiveness. If you apply in accordance with the law, you are requested to approve the application for this invention patent to encourage invention.

^a NCI 60 cell in vitro drug screening program with a concentration of 10 ^-5 M for each compound.

^b NT=No test.

^a NCI 60 cell in vitro drug screening program with a concentration of 10 ^-5 M for each compound.

^b NT=No test.

Claims (9)

a compound of formula (I), Wherein R is selected from the group consisting of: i) halo, amine, hydroxy and thio; ii) linear alkyl of NH(CH ₂ ) _n H, alkyl having a branch substituent An alkyl branch having an amino group substitution, an alkyl group having a hydroxyl group substitution, wherein 0 n 12; iii) O(CH ₂ ) _n H, N(CH ₃ ) ₂ , NH(CH ₂ ) _n NH(CH ₂ ) _n OH; wherein 0 n 10; iv) C _3-12 nitrogen-containing cycloalkyl and heterocyclic group containing from 1 to 3 heteroatoms selected from O, S and N, wherein the ortho, meta or para is more Independently selected from one of the following substituents: hydrogen, (CH ₂ ) _n alkyl, (CH ₂ ) _n hydroxy, (CH ₂ ) _n C _3-12 cycloalkyl, (CH ₂ ) _n C _{3- 12} nitrogen-containing cycloalkyl, (CH ₂ ) _n benzene ring, aldehyde group and (CH ₂ ) _n COC _3-12 nitrogen-containing cycloalkyl group; wherein n 10;v)NH(CH ₂ ) _n R ₁ ,0 n 10, R ₁ is selected from one of the following substituents: N(CH ₃ ) ₂ , C(NH ₂ ) ₂ , a linear alkyl group of NH(CH ₂ ) _n H , an alkyl group having a branched chain, An alkyl group having an amino group substitution, an alkyl group having a hydroxyl group substitution; vi) NH(CH ₂ ) _n R ₂ , 0 n 10, R ₂ is selected from one of the following substituents: a benzene ring, a C _3-12 cycloalkyl group and a heterocyclic group, containing from 1 to 3 hetero atoms selected from O, S and N, wherein the The position, the meta position or the para position may be independently selected from one of the following substituents: a methoxy group, an amine group, a benzene ring, an alkyl group having a C1-C3 branch substitution, an amine group, a nitro group, a hydroxyl group, and a C _3-12 heterocyclic ring; wherein the C _3-12 heterocyclic ring contains from 1 to 3 heteroatoms selected from O, S and N; and pharmaceutically acceptable salts, stereoisomers and mirror image isomers . The compound of claim 1, wherein the substituent group i)-vi) is selected from the group consisting of chlorine, hydroxyl, methoxy, dimethylamino, piperazin-1-yl, 4-methylpiperazine 1-yl, 4-ethylpiperazin-1-yl, 4-(2-hydroxyethyl)piperazin-1-yl, 4-benzylpiperazin-1-yl, 4-phenylpiperazine- 1-base, Morpholino, thiomorpholino, piperidin-1-yl, 4-hydroxypiperidin-1-yl, 4-benzylpiperidin-1-yl, [1,4'-bipiperidinyl]-1 '-yl, 4-(3-(piperidin-4-yl)propyl, pyrrolidin-1-yl, 2-oxopiperidin-1-yl, methylamino, ethylamino, propylamino, Butylamino, isobutyl, pent-3-ylamino, (2-(dimethylamino)ethyl)amino, (2-(diethylamino)ethyl)amino, 2-ethanolamino, 3- Propylamino, 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-indol-2-yl)amino, cyclohexyl, (1-benzylpiperidin-4- Amino, (thiophen-2-ylmethyl)amino, (cyclohexylmethyl)amino, benzylamino, (pyridin-2-ylmethyl)amino, (benzo[d][1,3] Olelen-5-ylmethyl)amino, (2-methoxybenzyl)amino, (3,4-dimethoxybenzyl)amino, phenethylamino, (4-methoxybenzene Ethyl)amino, (4 a group consisting of -aminophenethyl)amino, anthracene and piperidin-1-ylamino. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of 3-((4-chlorophenyl)thio)-2-hydroxyl Quinoline-4-carboxylic acid, 6,9-dichloro-12H-dehydrothiamine [2,3-c]quinolin-12-one, 10-chloro-6-hydroxy-12H-dehydrothiamine [2,3-c]quinolin-12-one, 10-chloro-6-methoxy-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6 -dimethylamino-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(piperazin-1-yl)-12H-dehydrothiamine and [2 ,3-c]quinolin-12-one, 10-chloro-6-(4-methylpiperazin-1-yl)-12H-dehydrothieno[2,3-c]quinolin-12- Ketone, 10-chloro-6-(4-ethylpiperazin-1-yl)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(4 -(2-hydroxyethyl)piperazin-1-yl)-12H-dehydrothieno[2,3-c]quinolinyl 12-one, 6-(4-benzylpiperazin-1-yl) -10-chloro-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(4-phenylpiperazin-1-yl)-12H-dehydrosulfur Amino[2,3-c]quinolin-12-one, 10-chloro-6-morpholino-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro -6-thiomorpholino-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-( Pyridine-1-yl)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(4-hydroxypiperidin-1-yl)-12H-dehydrogenation Thiamin[2,3-c]quinolin-12-one, 6-(4-benzylpiperidin-1-yl)-10-chloro-12H-dehydrothiamine[2,3-c] Quinoline-12-one, 6-([1,4'-bipiperidinyl]-1'-yl)-10-chloro-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6 -(4-(3-(piperidin-4-yl)propyl)piperidin-1-yl-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro- 6-(pyrrolidin-1-yl)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(2-oxopiperidin-1-yl) -12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-methylamino-12H-dehydrothieno[2,3-c]quinoline-12 -ketone, 10-chloro-6-ethylamino-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-propylamino-12H-dehydrothiamine And [2,3-c]quinolin-12-one, 6-(butylamino)-10-chloro-12H-dehydrothieno[2,3-c]quinolin-12-one, 10- Chloro-6-isobutyl-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(pent-3-ylamino)-12H-dehydrothiamine And [2,3-c]quinolin-12-one, 10-chloro-6-((2-(dimethylamino)ethyl)amino)-12H-dehydrothiamine[2,3-c Quinoline-12-one, 10-chloro-6-((2-(diethylamino)ethyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one , 10-chloro-6-(2-ethanolamino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(3-propanolamino)-12H -dehydrothiamine-[2,3-c]quinoline- 12-keto, 10-chloro-6-(5-pentanolamino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-((1-hydroxyl) Butan-2-yl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-((4-methylpentan-2-yl)amino )-12H-dehydrothieno[2,3-c]quinolin-12-one, 6-((2-aminoethyl)amino)-10-chloro-12H-dehydrothiamine [2, 3-c]quinolin-12-one, 10-chloro-6-((2-((2-hydroxyethyl))amino)ethyl)amino)-12H-dehydrothiamine[2,3-c Quinoline-12-one, 10-chloro-6-((2-morpholinoethyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10- Chloro-6-((3-(dimethylamino)propyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-((3) -(Diethylamino)propyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-((3-((2-hydroxyethyl)) Amino)propyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-((2,3-dihydro-1H-indole-2) -yl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(cyclohexyl)-12H-dehydrothiamine and [2,3- c] quinoline-12-one, 6-((1-benzylpiperidin-4-yl)amino)-10-chloro-12H-dehydrothieno[2,3-c]quinolin-12- ketone, 10-chloro-6-((thiophen-2-ylmethyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-((cyclohexyl) Methyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 6-(benzylamino)-10-chloro-12H-dehydrothiamine and [2,3 -c]quinolin-12-one, 10-chloro-6-((pyridin-2-ylmethyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 6-((Benzo[d][1,3]dioxol-5-ylmethyl)amino)-10-chloro-12H-dehydrothieno[2,3-c]quinoline -12-ketone, 10-chloro-6-((2-methoxybenzyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6- ((3,4-Dimethoxybenzyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(phenylethylamino)- 12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-((4-methoxyphenethyl)amino)-12H-dehydrothiamine and [2 ,3-c]quinolin-12-one, 6-((4-aminophenethyl)amino)-10-chloro-12H-dehydrothieno[2,3-c]quinolin-12-one , 2-(10-chloro-12-oxo-12H-dehydrothieno[2,3-c]quinolin-6-yl)indole, 10-chloro-6-(piperidin-1-ylamino) -12H-dehydrothieno[2,3-c]quinolin-12-one, and the salts thereof. A pharmaceutical composition comprising a therapeutically effective amount of a compound as claimed in claim 1 and at least one pharmaceutically acceptable carrier, diluent or excipient. A use of a compound according to any one of claims 1 to 4 for the manufacture of a medicament useful for inhibiting the activity of a first type of topoisomerase. A use of a compound according to any one of claims 1 to 4 for the manufacture of a medicament useful for inhibiting the activity of a second type of topoisomerase. A use of a compound according to any one of claims 1 to 4 for the manufacture of a medicament for the treatment of cancer. The use according to claim 7, wherein the cancer is selected from the group consisting of leukemia, non-small cell lung cancer, colorectal cancer, central nervous cancer, melanoma, ovarian cancer, kidney cancer, prostate cancer, and breast cancer. A method for producing a compound according to claim 1, which comprises: (1) mixing isatin, 2-((4-chlorophenyl)thio)acetic acid and sodium acetate, and reacting at 150 ° C for 1 hour. After the reaction mixture was cooled, acetic acid was added to obtain a precipitate, which was washed with acetic acid, water and n-hexane to obtain compound 2 (3-((4-chlorophenyl)thio)-2-hydroxyquinolin-4- carboxylic acid); (2) Take compound 2 (3-((4-chlorophenyl)thio)-2-hydroxyquinoline-4-carboxylic acid) dissolved in phosphorus oxychloride at 150 ° C for 48 hours, after cooling, the reaction is poured After entering the water, the precipitate was collected by filtration and placed in a 10% sodium hydrogen carbonate solution and stirred for 1 hour. The precipitate was collected and rinsed with water. The crude product was recrystallized from dichloromethane to give compound 3 (6,9-dichloro-12H-de Hydrogen thiamine [2,3-c]quinolin-12-one); (3) Compound 3 (6,9-dichloro-12H-dehydrothiamine [2,3-c]quinoline-12- The ketone) was dissolved in DMF, concentrated hydrochloric acid was added and refluxed for six hours, then concentrated hydrochloric acid was added dropwise and refluxed for further 12 hours. The reaction was dehydrated in vacuo and water was added. After filtration, the crude product was washed with ethanol to give compound 4 (10-chloro -6-hydroxy-12H-dehydrothieno[2,3-c]quinolin-12-one); (4) Suspension of compound 3 in methanol (6,9-dichloro-12H-dehydrogen sulfide) The amine [2,3-c]quinolin-12-one) was refluxed with sodium methoxide for 16 hours. After cooling, the solvent was removed, filtered and washed with ethanol and n-hexane to give compound 5 (10-chloro-6-methoxy -12H-dehydrothieno[2,3-c]quinolin-12-one); (5) Compound 3 (6,9-dichloro-12H-dehydrothiamine [2,3-c] Quinoline-12-one) with an appropriate secondary amine Dissolve with sodium carbonate in DMSO or DMF, reflux for 10 hours, and let stand for cooling. The reaction mixture is added to ice water, and the precipitate is filtered off, and then washed with water and methanol to obtain compound 6-21:10-chloro-6-di. Methylamino-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(piperazin-1-yl)-12H-dehydrothiamine and [2,3 -c]quinolin-12-one, 10-chloro-6-(4-methylpiperazin-1-yl)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(4-ethylpiperazin-1-yl)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(4-( 2-hydroxyethyl)piperazin-1-yl)-12H-dehydrothieno[2,3-c]quinolinyl 12-one, 6-(4-benzylpiperazin-1-yl)-10 -Chloro-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(4-phenylpiperazin-1-yl)-12H-dehydrothiamine [2,3-c]quinolin-12-one, 10-chloro-6-morpholino-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6 - thiomorpholino-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(piperidin-1-yl)-12H-dehydrothiamine [2,3-c]quinolin-12-one, 10-chloro-6-(4-hydroxypiperidin-1-yl)-12H-dehydrothieno[2,3-c]quinolin-12 -ketone, 6-(4-benzylpiperidin-1-yl)-10-chloro-12H-dehydrothiamine [2, 3-c]quinolin-12-one, 6-([1,4'-bipiperidinyl]-1'-yl)-10-chloro-12H-dehydrothieno[2,3-c]quina Porphyrin-12-one, 10-chloro-6-(4-(3-(piperidin-4-yl)propyl)piperidin-1-yl-12H-dehydrothieno[2,3-c]quinolin-12-one , 10-chloro-6-(pyrrolidin-1-yl)-12H-dehydrothieno[2,3-c]quinolin-12-one, and 10-chloro-6-(2-oxopiperine Pyridine-1-yl)-12H-dehydrothieno[2,3-c]quinolin-12-one; (6) compound 3 (6,9-dichloro-12H-dehydrothiamine [2] , 3-c]quinoline-12-one) is dissolved in DMSO with a suitable primary amine, refluxed for 8 hours, and after cooling, the reactant is added to 100 mL of water. The precipitate is filtered, washed with water and methanol to obtain compound N1. -N34: 10-Chloro-6-methylamino-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-ethylamino-12H-dehydrothiamine And [2,3-c]quinolin-12-one, 10-chloro-6-propylamino-12H-dehydrothieno[2,3-c]quinolin-12-one, 6-(butyl Benzyl)-10-chloro-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-isobutyl-12H-dehydrothiamine and [2,3 -c]quinolin-12-one, 10-chloro-6-(pent-3-ylamino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro- 6-((2-(Dimethylamino)ethyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-((2- Diethylamino)ethyl)amino)-12H- Hydrogenthio[2,3-c]quinolin-12-one, 10-chloro-6-(2-ethanolamino)-12H-dehydrothieno[2,3-c]quinolin-12- Ketone, 10-chloro-6-(3-propanolamino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(5-pentanolamino) -12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-((1-hydroxybutan-2-yl)amino)-12H-dehydrothiamine [2,3-c]quinolin-12-one, 10-chloro-6-((4-methylpentan-2-yl)amino)-12H-dehydrothieno[2,3-c]quina Porphyrin-12-one, 6-((2-aminoethyl)amino)-10-chloro-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6- ((2-((2-Hydroxyethyl)amino)ethyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-((2) -morpholinoethyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-((3-(dimethylamino)propyl) Amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-((3-(diethylamino)propyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(( 3-((2-hydroxyethyl)amino)propyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-((2,3) -dihydro-1H-indol-2-yl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-(cyclohexyl)-12H-de Hydrogenthio[2,3-c]quinolin-12-one, 6-((1-benzylpiperidin-4-yl)amino)-10-chloro-12H-dehydrothiamine [2, 3-c]quinolin-12-one, 10-chloro-6-((thiophen-2-ylmethyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one , 10-chloro-6-((cyclohexylmethyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 6-(benzylamino)-10-chloro- 12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-((pyridin-2-ylmethyl)amino)-12H-dehydrothiamine [2, 3-c]quinoline-12-one, 6-((benzo[d][1,3]dioxol-5-ylmethyl)amino)-10-chloro-12H-dehydrosulfur Amin[2,3-c]quinolin-12-one, 10-chloro-6-((2-methoxybenzyl)amino)-12H-dehydrothieno[2,3-c]quinoline -12-ketone, 10-chloro-6-((3,4-dimethoxybenzyl)amino)-12H-dehydrothieno[2,3-c]quinolin-12-one, 10- Chloro-6-(phenethylamino) -12H-dehydrothieno[2,3-c]quinolin-12-one, 10-chloro-6-((4-methoxyphenethyl)amino)-12H-dehydrothiamine and [ 2,3-c]quinolin-12-one, 6-((4-aminophenethyl)amino)-10-chloro-12H-dehydrothieno[2,3-c]quinolin-12- Ketone, 2-(10-chloro-12-oxo-12H-dehydrothieno[2,3-c]quinolin-6-yl)indole, and 10-chloro-6-(piperidin-1- Aminoamino)-12H-dehydrothieno[2,3-c]quinolin-12-one.