5-hydroxy-1,4-naphthalenedione for use in cancer treatment

Abstract

The present invention discloses compounds for inhibiting uncontrolled cell proliferation, particularly cancer stem cells. In particular, the present invention relates to compounds of formula (I) to formula (IV) for the treatment of cancer.

Description

Technical Field

[0001] The present invention relates to compounds for inhibiting uncontrolled cell proliferation, particularly for inhibiting cancer cells.

Background Art

[0002] Some relatively new anticancer agents act directly on abnormal proteins in cancer cells, which is called targeted therapy. Most chemotherapeutic drugs can be classified into alkylating agents, antimetabolites, anthracyclines, plant alkaloids, topoisomerase inhibitors, and other antitumor agents. Molecular targeted therapy is available as an expensive cancer therapy, but most of the world's population relies on standard chemotherapy.

[0003] Standard anticancer regimens target most of the dividing cancer cells and do not target cancer stem cells (CSC) that are in a quiescent state or have a slow division rate. Although it has been some time since CSCs were identified, scientists around the world are still aiming to discover CSC-targeted drugs, and unfortunately, to date, nothing specifically targeting CSCs is commercially available.

[0004] Therefore, it is important to develop CSC-specific therapeutic agents that effectively inhibit CSCs and act alone or in combination with standard therapies to provide effective treatment options for cancer patients.

Summary of the Invention

[0005] The present invention relates to compounds of formula I for treating various medical conditions, particularly for inhibiting uncontrolled cell proliferation or uncontrolled cell growth. The compounds are particularly effective against cancer cells. The compounds are also effective against cancer stem cells. The structure of formula I is as follows,

[0006]

Chemical Formula

Chemical formula

[0007] One aspect of the present invention includes a compound of formula II represented by the following structure. [ka]

[0008] One embodiment includes a compound of formula III represented by the following structure. [ka]

[0009] One aspect of the present invention includes a compound of formula IV represented by the following structure. [ka]

[0010] One aspect of the present invention relates to a pharmaceutical composition comprising the above-mentioned compound, at least one pharmaceutically acceptable additive, and optionally at least one active ingredient.

[0011] One aspect of the present invention relates to compounds of formulas I to IV for use in treating or inhibiting uncontrolled cell growth, such as cancer, including use targeting cancer cells such as cancer stem cells.

[0012] Another aspect of the present invention discloses a method for treating or inhibiting uncontrolled cell growth. This method comprises administering to a patient an effective amount of a compound of formulas I to IV or an effective amount of a pharmaceutical composition of any of formulas I to IV or the compounds described above. [Brief explanation of the drawing]

[0013] [Figure 1] The sphere analysis of the MDAMB231 cell line in the presence of the compound of Equation 1 and cisplatin is shown. [Figure 2] The sphere analysis of the PC3 cell line in the presence of the compound of Equation 1 and cisplatin is shown. [Figure 3] Sphere analysis of the MDAMB231 cell line in the presence of the compound of formula 2 and cisplatin is shown. [Figure 4] Sphere analysis of the PC3 cell line in the presence of the compound of Equation 2 and cisplatin is shown. [Figure 5] Sphere analysis of the MDAMB231 cell line in the presence of the compound of formula 7 and cisplatin is shown. [Figure 6] Sphere analysis of the PC3 cell line in the presence of the compound of Equation 7 and cisplatin is shown. [Figure 7] The sphere analysis of the MDAMB231 cell line in the presence of the compound of formula 37 and cisplatin is shown. [Figure 8]Sphere analysis of the PC3 cell line in the presence of the compound of formula 37 and cisplatin is shown. [Figure 9] Sphere analysis of the MDAMB231 cell line in the presence of the compound of formula 40 and cisplatin is shown. [Figure 10] Sphere analysis of the PC3 cell line in the presence of the compound of formula 40 and cisplatin is shown. [Figure 11] The sphere analysis of the MDAMB231 cell line in the presence of the compound of formula 41 and cisplatin is shown. [Figure 12] Sphere analysis of the PC3 cell line in the presence of the compound of formula 41 and cisplatin is shown. [Figure 13] The sphere analysis of the MDAMB231 cell line in the presence of the compound of formula 43 and cisplatin is shown. [Figure 14] The sphere analysis of the PC3 cell line in the presence of the compound of formula 43 and cisplatin is shown. [Figure 15] Sphere analysis of the MDAMB231 cell line in the presence of the compound of formula 46 and cisplatin is shown. [Figure 16] Sphere analysis of the PC3 cell line in the presence of the compound of formula 46 and cisplatin is shown. [Figure 17] Sphere analysis of the MDAMB231 cell line in the presence of the compound of formula 47 and cisplatin is shown. [Figure 18] Sphere analysis of the PC3 cell line in the presence of the compound of formula 47 and cisplatin is shown. [Figure 19] The sphere analysis of the MDAMB231 cell line in the presence of the compound of formula 52 and cisplatin is shown. [Figure 20] The sphere analysis of the PC3 cell line in the presence of the compound of formula 52 and cisplatin is shown. [Figure 21] The sphere analysis of the MDAMB231 cell line in the presence of the compound of formula 67 and cisplatin is shown. [Figure 22] The sphere analysis of the PC3 cell line in the presence of the compound of formula 67 and cisplatin is shown. [Figure 23] The sphere analysis of the MDAMB231 cell line in the presence of the compound of formula 68 and cisplatin is shown. [Figure 24] Sphere analysis of the PC3 cell line in the presence of the compound of formula 68 and cisplatin is shown. [Figure 25] The sphere analysis of the MDAMB231 cell line in the presence of the compound of formula 69 and cisplatin is shown. [Figure 26] Sphere analysis of the PC3 cell line in the presence of the compound of formula 69 and cisplatin is shown. [Figure 27] Sphere analysis of the MDAMB231 cell line in the presence of the compound of formula 70 and cisplatin is shown. [Figure 28] The sphere analysis of the PC3 cell line in the presence of the compound of formula 70 and cisplatin is shown. [Figure 29] Sphere analysis of the MDAMB231 cell line in the presence of the compound of formula 71 and cisplatin is shown. [Figure 30] The sphere analysis of the PC3 cell line in the presence of the compound of formula 71 and cisplatin is shown. [Figure 31] The sphere analysis of the MDAMB231 cell line in the presence of the compound of formula 72 and cisplatin is shown. [Figure 32] The sphere analysis of the PC3 cell line in the presence of the compound of formula 72 and cisplatin is shown. [Figure 33] The sphere analysis of the MDAMB231 cell line in the presence of the compound of formula 73 and cisplatin is shown. [Figure 34] Sphere analysis of the PC3 cell line in the presence of the compound of formula 73 and cisplatin is shown. [Figure 35] The sphere analysis of the MDAMB231 cell line in the presence of the compound of formula 74 and cisplatin is shown. [Figure 36] Sphere analysis of the PC3 cell line in the presence of compound 74 and cisplatin is shown. [Figure 37] The sphere analysis of the MDAMB231 cell line in the presence of the compound of formula 75 and cisplatin is shown. [Figure 38] The sphere analysis of the PC3 cell line in the presence of the compound of formula 75 and cisplatin is shown. [Figure 39] Sphere analysis of the MDAMB231 cell line in the presence of the compound of formula 76 and cisplatin is shown. [Figure 40] Sphere analysis of the PC3 cell line in the presence of the compound of formula 76 and cisplatin is shown. [Figure 41] The sphere analysis of the MDAMB231 cell line in the presence of the compound of formula 77 and cisplatin is shown. [Figure 42] The sphere analysis of the PC3 cell line in the presence of the compound of formula 77 and cisplatin is shown. [Figure 43] The sphere analysis of the MDAMB231 cell line in the presence of the compound of formula 78 and cisplatin is shown. [Figure 44] The sphere analysis of the PC3 cell line in the presence of the compound of formula 78 and cisplatin is shown. [Figure 45] The sphere analysis of the MDAMB231 cell line in the presence of the compound of formula 79 and cisplatin is shown. [Figure 46] Sphere analysis of the PC3 cell line in the presence of the compound of formula 79 and cisplatin is shown. [Figure 47] The sphere analysis of the MDAMB231 cell line in the presence of the compound of formula 80 and cisplatin is shown. [Figure 48] The sphere analysis of the PC3 cell line in the presence of the compound of formula 80 and cisplatin is shown. [Figure 49] The sphere analysis of the MDAMB231 cell line in the presence of the compound of formula 81 and cisplatin is shown. [Figure 50] The sphere analysis of the PC3 cell line in the presence of the compound of formula 81 and cisplatin is shown. [Figure 51] The activity of compounds Equations 2, 40, 41, 43, 52, 67, 68, 71, 72, and 73, as well as cisplatin, against the breast cancer MDAMB231 cell line in a soft agar assay is shown. [Figure 52] This figure shows the activity of compounds Equations 2, 40, 41, 43, 52, 67, 68, 71, 72, and 73, as well as cisplatin, against prostate cancer PC3 cell lines in a soft agar assay. [Figure 53] The activity of the compounds of formulas 1, 2, 40, 41, 43, 52, 67, 68, 69, 70, 71, 72, and 73 against lymphocytes is shown. [Figure 54] The wound-healing effects of compounds of formulas 2, 52, 40, and 43, as well as cisplatin, against breast cancer and prostate cancer are demonstrated. [Figure 55] The inhibitory effects of compounds with formulas 2, 52, and 40, as well as cisplatin, on aldehyde dehydrogenase (ALDH), a cancer stem cell (CSC) marker, are demonstrated. [Modes for carrying out the invention]

[0014] This invention relates to a compound of formula I for the treatment of various medical conditions, particularly for inhibiting uncontrolled cell growth or proliferation, or unregulated cell growth. The compound is particularly effective against cancer stem cells. The structure of the compound of formula I is as follows:

[0015] [ka] During the ceremony, n is between 1 and 10. Q is O, S, -NY' (where Y' is selected from -H and alkyl), R1, R2, R3, and R4 are each independently -H, alkoxy, alkyl, substituted or unsubstituted aromatic group, substituted or unsubstituted aromatic group having a condensed ring formed by a heterocycloalkyl group, -NH2, -NO2, -NHCOCH3, -CN, -O-, halogen, -OCF3, heterocycloalkyl group, -O-(CH2) n - Selected from heterocycloalkyl groups, R is a substituted or unsubstituted heterocycloalkyl, a substituted or unsubstituted cycloalkyl, -NR 10 R 11 , -NR 10 R 11 • HCl or acid salt, -OR 10 R 11 ,-CONR 10 R 11 , -NR 10 R 11 CONR 10 R 11 , -NR 10 R 11 SOONR 10 R 11 Selected from -COOH, (In the above formula, R 10 and R 11 Each of these is independently selected from -H, alkyl, substituted or unsubstituted aryl, heteroaryl, alkylamine, substituted arylamine, substituted or unsubstituted cycloalkyl, -CH2-CH2-O-alkyl, or R 10 and R 11 They combine to form substituted or unsubstituted cycloalkyl or heterocycloalkyl groups, or R 10 and R 11 They combine to form a substituted or unsubstituted cycloalkyl or heterocycloalkyl ring with -N included in the ring, or R 10 It is expressed by the following formula: [ka] In the above formula, R 13 The group is selected from -OH, -NH2, -NHCOCH3, alkyl, acetyl, C3-C8 acyl group, and X (selected from F, Cl, Br). R 14 The group is selected from alkoxy, -OMe, -OH, NH2, -NHCOCH3, alkyl, acetyl, C3-C8 acyl group, and X (selected from F, Cl, Br). R 15The group is selected from alkoxy, -OMe, -OH, -H, Br, NH2, alkyl, acetyl, C3-C8 acyl group, and X (selected from F, Cl, Br). R 16 (Selected from -H, -CH2OH, -OH, alkyl, and alkoxy) and R6 is selected from the R group, -H, and the following formula as defined above. [ka] (R5 is located at any position, exists as one or more groups, and is selected from -CH2-O-CH2, -COOH, alkyl, alkoxy, NHCOCH3, -H, -OR, -NR, X (selected from F, Cl, Br), or R5 forms a condensed ring having an -O-CH2-O- group.)

[0016] One embodiment of the present invention discloses a compound represented by the following formula II.

[0017] [ka]

[0018] In one embodiment of the present invention, the compound of formula III is expressed as follows: [ka]

[0019] In one embodiment of the present invention, the compound of formula IV is expressed as follows: [ka]

[0020] In one embodiment of the present invention, the compound comprises the following: R1, R2, and R3 are each independently selected from -H. R4 is a selected -H, alkoxy, alkyl, substituted or unsubstituted aromatic group, -NH2, -NO2, -NHCOCH3, -CN, -O-, halogen, -OCF3, [ka] And, R6 is -H, [ka] Selected from, Q is selected from -O and -NH. R is -COOH, [ka] Selected from, n is between 1 and 6. * This represents a connection point.

[0021] In one embodiment of the present invention, the compound comprises the following: -Q-(CH2) n - The R group is not present. R1, R2, and R3 are each independently selected from -H, and R6 is -H. R4 is [ka] Selected from, * represents a connection point.

[0022] The compounds included in formulas I to IV are as follows:

[0023] [ka]

[0024] [ka]

[0025] [ka]

[0026] [ka]

[0027] The present invention also encompasses pharmaceutical compositions comprising a compound of formulas I to IV or any of the above compounds, at least one pharmaceutically acceptable additive, and optionally at least one active ingredient.

[0028] The active ingredients are not limited to those listed below, but are selected from any combination of the following ingredients. Imatinib, nilotinib, gefitinib, sunitinib, carfilzomib, salinosporamide A, retinoic acid, cisplatin, carboplatin, oxaliplatin, mechloretamine, cyclophosphamide, chlorambutyl, ifosfamide, azathioprine, mercaptopurine, doxifluridine, fluorouracil, gemcitabine, methotrexate, thioguanine, vincristine, vinblastine, vinorelbine, vin Decine, podophyllotoxin, etoposide, teniposide, tafluposide, paclitaxel, docetaxel, irinotecan, topotecan, amsacrin, actinomycin, doxorubicin, daunorubicin, barurubicin, idarubicin, epirubicin, plicamycin, mitomycin, mitoxantrone, melphalan, busulfan, capecitabine, pemetrexed, eposilon, 13-cis-retinoic acid, 2-CdA , 2-chlorodeoxyadenosine, 5-azacitidine, 5-fluorouracil, 5-FU, 6-mercaptopurine, 6-MP, 6-TG, 6-thioguanine, Abraxane, Accutane (registered trademark), Actinomycin-D, Adriamycin (registered trademark), Adolsil (registered trademark), Afinitor (registered trademark), Agrylin (registered trademark), Ala-Cort (registered trademark), Aldesleukin, Alemtuzumab, Alimta, Alitretinoin, Alkaban-AQ (registered trademark), Alkeran (registered trademark), Trans retinoic acid, Alpha interferon, Altretamine, Ametopterin, Amiphostin, Aminoglutethimide, Anagrelide, Arimidex (registered trademark), Aromasin (registered trademark), Alanone (registered trademark), Arsenic trioxide, Arzerra (trademark), Asparaginase, ATRA, Avastin (registered trademark), Azacitidine, BCG, BCNU, bendamustine, bevacizumab, bexarotene, BEXXAR®, bicalutamide, BiCNU, Blenoxane®, bleomycin, bortezomib, busulfan, busulfex®, C225, leucovorin calcium, Campath®, Camptosar®, camptothecin-11, capecitabine, Carac®, carboplatin, carmustine, carmustine wafer, Casodex®, CC-5013, CCI-779, CCNU, CDDP, CeeNU, Serubidin®, Cetuximab, Chlorambucil, Citrobolum Factor, Cladribine, Cortisone, Cosmegen®, CPT-11, Sitadren®, Cytosar-U®, Cytoxan®, Dacarbazine, Dacogen, Dactinomycin, Darbepoetin alfa, Dasatinib, Daunomycin, Daunorubicin Hydrochloride, Daunorubicin Liposome, Daunoxosome®, Decadron, Decitabine, Delta-Cortef®, Deltason®, Denileukin Difutitex, DepoCyt®, Dexamethasone, Dexamethasone Acetate, Dexamethasone Sodium Phosphate, Dexasone, Dexrazoxane, DHAD, DIC, Diodex, Docetaxel, Doxil®, Doxorubicin, Doxorubicin Liposome, Droxia®, DTIC, DTIC-Dome®, Duralone®, FDEX®, Eliger D (trademark), Ellence (trademark), Eloxatin (trademark), Elspar (registered trademark), Emcyt (registered trademark), Epirubicin, Epoetin alfa, Elbitax, Erlotinib, Erwinia L-asparaginase, Estramustine, Ethyol, Etopophos (registered trademark), Etoposide, Etoposide phosphate, Eulexin (registered trademark), Everolimus, Evista (registered trademark), Exemestane, Fareston®, Faslodex®, Femara®, Filgrastim, Floroxuridine, Fludara®, Fludarabine, Fluoroplex®, Fluorouracil, Fluorouracil (cream), Fluoxymesterone, Flutamide, Folic Acid, FUDR®, Fulvestrant, G-CSF, Gefitinib, Gemcitabine, Gemtuzumab, Ozogamicin, Gemzar Gleevec (trademark), Gliadel (registered trademark) wafer, GM-CSF, Goserelin, Granulocyte colony-stimulating factor, Granulocyte-macrophage colony-stimulating factor, Halotestin (registered trademark), Herceptin (registered trademark), Hexadrol, Hexalen (registered trademark), Hexamethylmelamine, HMM, Hycamtin (registered trademark), Hydrea (registered trademark), Hydrocort Acetate (registered trademark), Hydrocortisone, Hydrocortisone sodium phosphate, Hydrocortisone sodium succinate, Hydrocortisone phosphate, Hydroxyurea, Ibritumomab, Ibritumomab, Tiuxetan, Idarubicin (registered trademark), Idarubicin Ifex(registered trademark), IFN-Alpha, Ifosfamide, IL-11, IL-2, Imatinib Mesylate, Imidazole Carboxamide, Interferon Alpha, Interferon Alpha-2b (PEG Conjugate), Interleukin-2, Interleukin-11, Intron A(registered trademark) (Interferon Alpha-2b), Iressa(registered trademark), Irinotecan, Isotretinoin, Ixabepyrone, Ixempra(trademark), Kidrolase(registered trademark), Lanacort(registered trademark), Lapatinib, L-Asparaginase, LCR, Lenalidomide, Letrozole, Leucovorin, Leukeran, Leukine(trademark), Leuprolide, Leulocristine, Leustatin(trademark), Liposome Ara-C, Liquid Pred(registered trademark), Lomustine, L-PAM, L-Sarcolicin, Lupron(registered trademark), Lupron Depot (registered trademark), Matulane®, Maxidex, Mechloretamine, Mechloretamine Hydrochloride, Medralone®, Medrol®, Megace®, Megestrol, Megestrol Acetate, Melphalan, Mercaptopurine, Mesna, Mesnex®, Methotrexate, Methotrexate Sodium, Methylprednisolone, Meticorten®, Mitomycin, Mitomycin-C, Mitozantrone, M-Prednisol®, MTC MTX, Mustargen (registered trademark), Mustine, Mutamycin (registered trademark), Myleran (registered trademark), Mylocel (trademark), Mylotarg (registered trademark), Navelbine (registered trademark), Nelarabine, Neosar (registered trademark), Neulasta (trademark), NeμMega (registered trademark), Newpogen (registered trademark), Nexavar (registered trademark), Nilandron (registered trademark), Nilotinib, Niltamide, Nipent (registered trademark), Nitrogen Mustard, Novaldex (registered trademark) (Trademark), Novantrone (Registered Trademark), Nplate, Octreotide, Octreotide Acetate, Ofatumumab, Oncospar (Registered Trademark), Oncovin (Registered Trademark), Ontak (Registered Trademark), Onxal (Trademark), Oprelbequin, Orapred (Registered Trademark), Orasone (Registered Trademark), Oxaliplatin, Palicaxel, Protein-bound Palicaxel, Pamidronate, Panitumumab, Panretin (Registered Trademark), Paraplatin (Registered Trademark), Pazopanib, Pediapred (Registered Trademark) (Registered Trademark), PEG Interferon, Pegaspargase, Pegfilgrastim, PEG-INTRON (Trademark), PEG-L-Asparaginase, Pemetrexed, Pentostatin, Phenylalanine Mustard, Platinol (Registered Trademark), Platinol-AQ (Registered Trademark), Prednisolone, Prednisone, Prelone (Registered Trademark), Procarbazine, PROCRIT (Registered Trademark), Proleukin (Registered Trademark), Prolifeprospan 20 with Carmustine Implant, Purinethol (Registered Trademark), Raloxifene, Revlimid®, Rheumatrex®, Rituxan®, Rituximab, Loferon-A® (Interferon Alpha 2a), Romiplostim, Rubex®, Rubidomycin Hydrochloride, Sandostatin®, Sansostatin LAR (registered trademark), Sarglamostim, Solu-Cortef (registered trademark), Solu-Medrol (registered trademark), Sorafenib, SPRYCEL (trademark), STI-571, Streptozocin, SU11248, Sunitinib, Sutent (registered trademark), Tamoxifen, Tarceva (registered trademark), Targretin (registered trademark), Tasigna (registered trademark), Taxol (registered trademark), Taxotere (registered trademark), Temodar (registered trademark), Temozolomide, Temsirolimus, Teniposide, TESPA, Thalidomide, Salomide (registered trademark), TheraCys (registered trademark), Thioguanine, Thioguanine Tabloid (registered trademark) ), Thiophosphoamide, Thioplex(registered trademark), Thiotepa, TICE(registered trademark), Toposar(registered trademark), Topotecan, Toremifene, Torisel(registered trademark), Tositumomab, Trastuzumab, Treanda(registered trademark), Tretinoin, Trexall(trademark), Trisenox(registered trademark), TSPA, TYKERB(registered trademark), VCR, Vectibix(trademark), Velban(registered trademark), Velcade(registered trademark), VePesid(registered trademark), Vesanoid(registered trademark), Viadur(trademark), Vidaza(registered trademark), Vinblastine, Vinblastine sulfate, Vincasar Pfs(registered trademark), vincristine, vinorelbine, vinorelbine tartrate, VLB, VM-26, vorinostat, votrient, VP-16, VμMon(registered trademark), Xeloda(registered trademark), Zanosar(registered trademark), Zevalin(trademark), Zinecard(registered trademark), Zoladex(registered trademark), Zoledronic acid, Zolinza, Zometa(registered trademark).

[0029] Pharmaceutically acceptable additives include carriers, adjuvants, vehicles, or mixtures thereof.

[0030] The compounds of the present invention are used to treat or inhibit uncontrolled cell growth, such as in cancer. These compounds effectively target cancer cells, including cancer stem cells.

[0031] The present invention also relates to a method for treating or inhibiting uncontrolled cell growth, such as cancer. The compound has been found to target cancer cells, including cancer stem cells. The method involves administering an effective amount of one or more compounds of formulas I to IV to a patient.

[0032] The present invention also relates to a method for treating or inhibiting uncontrolled cell growth, such as cancer, by administering to a patient an effective amount of a pharmaceutical composition containing one or more compounds of formulas I to IV, or any of the above compounds.

[0033] The compounds of the present invention may also be provided in conjunction with standard therapies available for the treatment of cancer.

[0034] The compounds of the present invention are used for the treatment or inhibition of at least one of the following cancers: breast cancer, prostate cancer, brain cancer, blood cancer, bone marrow cancer, liver cancer, pancreatic cancer, skin cancer, kidney cancer, colon cancer, ovarian cancer, lung cancer, testicular cancer, penile cancer, thyroid cancer, parathyroid cancer, pituitary cancer, thymic cancer, retinal cancer, uveal cancer, conjunctival cancer, spleen cancer, head cancer, neck cancer, tracheal cancer, gallbladder cancer, rectal cancer, salivary gland cancer, adrenal gland cancer, throat cancer, esophageal cancer, lymph node cancer, sweat gland cancer, sebaceous gland cancer, muscle cancer, heart cancer, and gastric cancer. In particular, the compounds are used for the treatment of breast cancer and prostate cancer.

[0035] The compound was found to have lower activity against normal cells (lymphocytes) compared to its activity against cancer cells.

[0036] The compound was found to have wound-healing effects in breast cancer and prostate cancer.

[0037] The compound was found to inhibit aldehyde dehydrogenase (ALDH), a cancer stem cell (CSC) marker.

[0038] In one embodiment, the compound can be used to treat malaria and dengue fever.

[0039] The synthesis method for the compound is described below. [Examples]

[0040] The embodiments shown below are intended to clarify, rather than limit, the present invention.

[0041] Scheme 1: [ka] Reagents and conditions: a. Acetic anhydride, pyridine, room temperature, 12 hours; b. NBS, AcOH, H2O, 65°C, 2 hours; c. 5N H2SO4, retarder, 90°C, 2 hours.

[0042] Synthesis of compound 2 (1,5-diacetate naphthalene): In a clean, dry three-necked round-bottom flask (RB), 1,5-dihydroxynaphthalene (20 g, 0.1249 mol) was added to pyridine (100 mL), and the reaction mixture was stirred at room temperature (RT) for 15 minutes. The temperature was then reduced to 0°C. Weighed acetic anhydride (57.28 gm, 0.5620 mol) was added dropwise to the RM at 0°C, and the reaction mixture was stirred for 12 hours, monitored using TLC. The reaction mixture was slowly poured into ice-cold water (1000 mL) and stirred. The reaction mixture was stirred using an overhead stirrer for 45 minutes. The reaction mixture was filtered, and the precipitate was dissolved in MDC (1000 mL). The organic layer was washed with copper sulfate solution (250 mL x 5 times) and brine solution (200 mL x 3 times). The reaction mixture was concentrated under reduced pressure. The resulting crude compound was purified by simple filtration column chromatography (hexane:ethyl acetate, 40:60). Pure compound = 24 gm. Yield = 79%.

[0043] 1H NMR(CDCl3,400MHz):δ=7.77(dd,J=8.5Hz,2H),7.49(t,J=8.0Hz,2H),7.28(d,J=7.5Hz,2H),2.44(s,6H)

[0044] Synthesis of compound 3 (2-bromo-1,4-dihydro-1,4-dioxonaphthalene-5-ylacetate): In a clean, dry three-necked RB, NBS (58.07 gm, 0.3277 mol) was added in water (500 mL) and acetic acid (500 mL), and the reaction mixture was incubated at 45°C for 15 minutes. Compound 1 (20 gm, 0.0819 mol) was dissolved in acetic acid (500 mL) and heated at 45°C. The solution of Compound 1 was added dropwise to the NBS reaction mixture at 45°C over 30 minutes. The reaction mixture was stirred at 45°C for 40 minutes. The temperature was raised to 65°C and stirred for 1 hour. The reaction mixture was monitored by TLC. After completion, the reaction mixture was cooled to room temperature, poured into water (1500 mL), and extracted with MDC (250 mL x 6 times). The organic layer was washed with saturated sodium bicarbonate and brine aqueous solution (200 mL x 3 times). The combined organic layers were vacuum-dried and concentrated under reduced pressure. Crude compound = 33 gm.

[0045] 1 H NMR(CDCl3,400MHz):δ=8.15(dd,J=1.2,8.0Hz,1H),7.77(t,J=8.0Hz,1H),7.42(dd,J=1.2,8.0Hz,1H),7.38(s,1H),2.44(s,3H)

[0046] Synthesis of compound 4 (2-bromo-5-hydroxynaphthalene-1,4-dione): Compound 3, dissolved in retarder (715 mL) at 45°C for 15 minutes, was added to a clean, dry three-necked RB and stirred. Then, 5N sulfuric acid (396 mL) was slowly added. The reaction mixture was refluxed at 90°C for 2 hours. The reaction was monitored by TLC. The reaction mixture was evaporated to dryness on a rotary evaporator. The reaction mixture was poured into 1000 mL of water and extracted by MDC (250 mL x 6 times). The organic layer was washed with brine solution (200 mL x 3 times) and dried on sodium sulfate. The organic layer was concentrated under reduced pressure. The resulting crude compound was purified by simple filtration column chromatography (hexane:ethyl acetate, 80:20). Pure compound yielded 9.9 gm. Yield 35%.

[0047] 1 H NMR(CDCl3,400MHz):δ=11.81(s,1H),7.73(d,J=8.2Hz,1H),7.67(t,J=8.2Hz,1H),7.32(d,J=8.2Hz,1H),7.20(s,1H)

[0048] Scheme 2: [ka] Reagents and conditions: a. Substituted phenylboronic acid, Pd(PPh3)4, Na2CO3, THF, water, room temperature, 12 hours; b. 4-(2-chloroethyl)morpholine hydrochloride, K2CO3, DMF, 100°C, 3 hours

[0049] Synthesis of compound 5a (5-hydroxy-2-(4-methoxyphenyl)naphthalene-1,4-dione): Compound 4 (1.0 g, 39.2 mmol) and 4-methoxyphenylboronic acid (0.72 g, 47.4 mmol) were dissolved in THF (108 mL) and water (12 mL), to which Na2CO3 (0.82 g, 78.4 mmol) was added to the reaction mixture. Pd(PPh3)4 (0.226 g, 1.97 mmol) was added under a nitrogen atmosphere, and the mixture was stirred at room temperature for 30 minutes. The reaction mixture was stirred at room temperature for 16 hours and monitored using TLC. After completion, the reaction mixture was cooled to room temperature, poured into water (200 mL), and extracted with ethyl acetate (100 mL x 3 times). The organic layer was washed with water (100 mL). The combined organic layer was dried on anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude compound was purified by column chromatography (hexane:ethyl acetate, 90:10). Pure compound = 0.45 g / m. Yield = 45%.

[0050] 1 H NMR(CDCl3,400MHz):δ=12.07(s,1H),7.71(d,J=8.2Hz,1H),7.69(d,J=8.2Hz,1H),7.6 6(d,J=8.0Hz,1H),7.59(d,J=2.4Hz,1H),7.29(d,J=1.2Hz,1H),7.0(m,3H),3.87(s,3H)

[0051] Synthesis of compound 5b (2-(4-fluorophenyl)-5-hydroxynaphthalene-1,4-dione): Compound 4 (1.0 g, 39.2 mmol) and 4-fluorophenylboronic acid (0.66 g, 47.4 mmol) were dissolved in THF (108 mL) and water (12 mL), to which Na2CO3 (0.82 g, 78.4 mmol) was added to the reaction mixture. Pd(PPh3)4 (0.226 g, 1.97 mmol) was added under a nitrogen atmosphere, and the mixture was stirred at room temperature for 30 minutes. The reaction mixture was stirred at room temperature for 16 hours and monitored using TLC. After completion, the reaction mixture was cooled to room temperature, poured into water (200 mL), and extracted with ethyl acetate (100 mL x 3 times). The organic layer was washed with water (100 mL). The combined organic layer was dried on anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude compound was purified by column chromatography (hexane:ethyl acetate, 90:10). Pure compound = 0.40 g / m. Yield = 41%.

[0052] 1 H NMR(CDCl3,400MHz):δ=11.99(s,1H),7.72(d,J=6.0Hz,1H),7.68(d,J=6.0Hz,1H),7.60(m,2H),7.31(d,J=6.8Hz,1H),7.19(m,2H),7.02(s,1H)

[0053] Synthesis of compound 5c(2-(benzo[d][1,3]dioxol-6-yl)-5-hydroxynaphthalene-1,4-dione): Compound 4 (1.0 g, 39.2 mmol) and 3,4(methylenedioxy)phenylboronic acid (0.65 g, 39.2 mmol) were dissolved in THF (90 mL) and water (10 mL), to which Na2CO3 (0.83 g, 78.4 mmol) was added to the reaction mixture. Pd(PPh3)4 (0.226 g, 1.96 mmol) was added under a nitrogen atmosphere, and the mixture was stirred at room temperature for 30 minutes. The reaction mixture was stirred at room temperature for 16 hours and monitored using TLC. After completion, the reaction mixture was cooled to room temperature, poured into water (200 mL), and extracted with ethyl acetate (100 mL x 3 times). The organic layer was washed with water (100 mL). The combined organic layer was dried on anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude compound was purified by column chromatography (hexane:ethyl acetate, 90:10). Pure compound = 0.72 g / m. Yield = 64%.

[0054] 1 H NMR(CDCl3,400MHz):δ=12.03(s,1H),7.71(d,J=8.2Hz,2H),7.69(d,J=8.2Hz,1H), 7.30(d,J=8.2Hz,1H),7.14(m,2H),6.98(s,1H),6.91(d,J=8.0Hz,1H),6.04(s,2H)

[0055] Synthesis of the compound (5-(2-morpholinoethoxy)-2-(4-fluorophenyl)naphthalene-1,4-dione) of formula 7: Compound 5b (0.25 gm, 9.36 mmol) and DMF (20 mL) were added to a two-necked RBF (100 mL). K2CO3 (0.26 g, 18.7 mmol) and KI (0.015 gm, 0.93 mmol) were added to the reaction mixture and stirred at room temperature for 15 minutes. 4-(2-chloroethyl)morpholine hydrochloride (0.209 gm, 11.23 mmol) was added to the reaction mixture. The reaction mixture was heated at 100 °C for 4 hours. The reaction was monitored by TLC. After completion, the reaction mixture was cooled to room temperature, poured into water (100 mL), and extracted with ethyl acetate (100 mL x 3 times). The organic layer was washed with water (100 mL x 3). The combined organic layer was dried on anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude compound was purified by column chromatography (ethyl acetate:methanol, 95:5). Pure compound = 206 mg. Yield = 58%.

[0056] 1 H NMR(CDCl3,400MHz):δ=7.84(d,J=6.0Hz,1H),7.71(d,J=6.0Hz,1H),7.58(m,2H),7.33(d,J=6.8Hz, 1H),7.17(m,2H),6.93(s,1H),4.31(t,J=4.4Hz,2H),3.75(m,4H),2.98(t,J=4.8Hz,2H),2.72(m,4H)

[0057] Synthesis of the compound (5-(2-morpholinoethoxy)-2-(benzo[d][1,3]dioxol-6-yl)naphthalene-1,4-dione) of formula 1: Compound 5c (0.2 gm, 8.40 mmol) and DMF (20 mL) were added to a two-necked RBF (100 mL). K2CO3 (0.23 g, 16.8 mmol) and KI (0.013 gm, 0.84 mmol) were added to the reaction mixture and stirred at room temperature for 15 minutes. 4-(2-chloroethyl)morpholine hydrochloride (0.187 gm, 10.08 mmol) was added to the reaction mixture. The reaction mixture was heated at 100 °C for 4 hours. The reaction was monitored by TLC. After completion, the reaction mixture was cooled to room temperature, poured into water (100 mL), and extracted with ethyl acetate (100 mL x 3 times). The organic layer was washed with water (100 mL x 3). The combined organic layer was dried on anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude compound was purified by column chromatography (ethyl acetate:methanol, 95:5). Pure compound = 40 mg. Yield = 16%.

[0058] 1 H NMR(CDCl3,400MHz):δ=7.83(d,J=8.0Hz,1H),7.69(d,J=8.2Hz,1H),7.31(d,J=8.2Hz,1H),7.11(m, 2H),6.90(m,2H),6.02(s,2H),4.32(t,J=4.4Hz,2H),3.78(m,4H),3.04(t,J=4.8Hz,2H),2.82(m,4H)

[0059] Scheme 3: [ka] Reagents and conditions: a. K2CO3, DMF, room temperature, 4 hours; b. 4-(2-chloroethyl)morpholine hydrochloride, K2CO3, DMF, 100°C, 4 hours

[0060] Synthesis of compound 7a (2-(4-methoxyphenoxy)-5-hydroxynaphthalene-1,4-dione) 4-methoxyphenol (0.972 gm, 78.4 mmol) and DMF (50 mL) were added to a two-necked RBF (250 mL). K2CO3 (1.08 g, 78.4 mmol) was added to the reaction mixture and stirred at room temperature for 15 minutes. Compound 4 (2.0 gm, 78.4 mmol) was added to the reaction mixture. The reaction mixture was stirred at room temperature for 3 hours. The reaction was monitored by TLC. After completion, the reaction mixture was poured into water (200 mL) and extracted with ethyl acetate (100 mL x 3 times). The organic layer was washed with water (100 mL x 3). The combined organic layer was dried on anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude compound was purified by column chromatography (petroleum ether:ethyl acetate, 95:5). Pure compound = 0.538 gm. Yield = 24%.

[0061] 1 H NMR(CDCl3,400MHz):δ=12.07(s,1H),7.86(d,J=8.0Hz,1H),7.80(d,J=0.8Hz,1H ),7.65(d,J=0.8Hz,1H),7.20(d,J=9.2Hz,2H),7.07(d,J=9.2Hz,2H),5.60(s,1H)

[0062] Synthesis of compound 7b (2-(4-fluorophenoxy)-5-hydroxynaphthalene-1,4-dione): 4-fluorophenol (0.443 gm, 39.5 mmol) and DMF (50 mL) were added to a two-necked RBF (100 mL). K2CO3 (0.54 g, 39.5 mmol) was added to the reaction mixture and stirred at room temperature for 15 minutes. Compound 4 (1.0 gm, 39.5 mmol) was added to the reaction mixture. The reaction mixture was stirred at room temperature for 3 hours. The reaction was monitored by TLC. After completion, the reaction mixture was poured into water (200 mL) and extracted with ethyl acetate (100 mL x 3 times). The organic layer was washed with water (100 mL x 3). The combined organic layer was dried on anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude compound was purified by column chromatography (petroleum ether:ethyl acetate, 95:5). Pure compound = 1.05 gm. Yield = 78%.

[0063] 1 H NMR(CDCl3,400MHz):δ=12.07(s,1H),7.74(d,J=1.2Hz,1H),7.62(m,1H),7.31(dd,J=1.2Hz & 7.6Hz,1H),7.17(m,4H),5.87(s,1H)

[0064] Synthesis of compound 7c(2-(benzo[d][1,3]dioxol-5-yloxy)-5-hydroxynaphthalene-1,4-dione): Sesamol (1.08 gm, 78.4 mmol) and DMF (50 mL) were added to a two-necked RBF (100 mL). K2CO3 (0.54 g, 78.4 mmol) was added to the reaction mixture and stirred at room temperature for 15 minutes. Compound 4 (2.0 gm, 78.4 mmol) was added to the reaction mixture. The reaction mixture was stirred at room temperature for 4 hours. The reaction was monitored by TLC. After completion, the reaction mixture was poured into water (200 mL) and extracted with ethyl acetate (100 mL x 3 times). The organic layer was washed with water (100 mL x 3). The combined organic layer was dried on anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude compound was purified by column chromatography (petroleum ether:ethyl acetate, 95:5). Pure compound = 0.676 gm. Yield = 25%.

[0065] 1 H NMR(CDCl3,400MHz):δ=12.11(s,1H),7.74(d,J=0.8Hz,1H),7.72(dd,J=1.2Hz,8.4Hz,1H),7.30(d, J=0.8Hz,1H),6.85(d,J=8Hz,1H),6.62(dd,J=2.4Hz,7.6Hz,1H),6.58(d,J=2.4Hz,1H),6.05(s,2H)

[0066] Synthesis of the compound (5-(2-morpholinoethoxy)-2-(4-methoxyphenoxy)naphthalene-1,4-dione) of formula 43 Compound 7a (0.32 gm, 10.94 mmol) and DMF (20 mL) were added to a two-necked RBF (100 mL). K2CO3 (0.30 g, 21.9 mmol) was added to the reaction mixture and stirred at room temperature for 15 minutes. 4-(2-chloroethyl)morpholine hydrochloride (0.41 gm, 21.9 mmol) was added to the reaction mixture. The reaction mixture was heated at 100 °C for 6 hours. The reaction was monitored by TLC. After completion, the reaction mixture was cooled to room temperature, poured into water (100 mL), and extracted with ethyl acetate (100 mL x 3 times). The organic layer was washed with water (100 mL x 3). The combined organic layer was dried on anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude compound was purified by column chromatography (ethyl acetate:methanol, 95:5). Pure compound = 69 mg. Yield = 21%.

[0067] 1 H NMR (CDCl3,400MHz): δ=7.88(d,J=6.8Hz,1H),7.68(d,J=8.4Hz,1H),7.34(d,J=8.4Hz,1H),7.17(m, 4H),5.82(s,1H),4.27(t,J=5.6Hz,2H),3.83(s,3H),3.72(m,4H),2.92(t,J=5.6Hz,2H),2.68(m,4H)

[0068] Synthesis of the compound (5-(2-morpholinoethoxy)-2-(4-fluorophenoxy)naphthalene-1,4-dione) of formula 75 Compound 7b (0.32 gm, 10.94 mmol) and DMF (20 mL) were added to a two-necked RBF (100 mL). K2CO3 (0.30 g, 21.9 mmol) was added to the reaction mixture and stirred at room temperature for 15 minutes. 4-(2-chloroethyl)morpholine hydrochloride (0.41 gm, 21.9 mmol) was added to the reaction mixture. The reaction mixture was heated at 100 °C for 6 hours. The reaction was monitored by TLC. After completion, the reaction mixture was cooled to room temperature, poured into water (100 mL), and extracted with ethyl acetate (100 mL x 3 times). The organic layer was washed with water (100 mL x 3). The combined organic layer was dried on anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude compound was purified by column chromatography (ethyl acetate:methanol, 95:5). Pure compound = 69 mg. Yield = 21%.

[0069] 1 H NMR(CDCl3,400MHz):δ=7.78(d,J=1.2Hz,1H),7.62(m,1H),7.31(dd,J=1.2Hz & 7.6Hz,1H),7.17(m,4H),5.82(s,1H),4.27(t,J=5.6Hz,2H),3.72(m,4H),2.92(t,J=5.6Hz,2H),2.68(m,4H)

[0070] Synthesis of compound formula 46 (5-(2-morpholinoethoxy)-2-(benzo[d][1,3]dioxol-5-yloxy)naphthalene-1,4-dione): Compound 7c (0.5 gm, 17.66 mmol) and DMF (20 mL) were added to a two-necked RBF (100 mL). K2CO3 (0.49 g, 35.3 mmol) was added to the reaction mixture and stirred at room temperature for 15 minutes. 4-(2-chloroethyl)morpholine hydrochloride (0.395 gm, 21.2 mmol) was added to the reaction mixture. The reaction mixture was heated at 100 °C for 2 hours. The reaction was monitored by TLC. After completion, the reaction mixture was cooled to room temperature, poured into water (100 mL), and extracted with ethyl acetate (100 mL x 3 times). The organic layer was washed with water (100 mL x 3). The combined organic layer was dried on anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude compound was purified by column chromatography (ethyl acetate:methanol, 95:5). Pure compound = 294 mg. Yield = 42%.

[0071] 1 H NMR (CDCl3,400MHz): δ=7.74(d,J=0.8Hz,1H),7.72(dd,J=1.2Hz,8.4Hz,1H),7.30(d,J=0.8Hz,1H),6.85(d,J=8Hz,1H),6.62(dd,J=2. 4Hz,7.6Hz,1H),6.58(d,J=2.4Hz,1H),6.05(s,2H),5.91(s,1H),4.27(t,J=5.6Hz,2H),3.72(m,4H),2.92(t,J=5.6Hz,2H),2.68(m,4H)

[0072] Scheme 4: [ka] Reagents and conditions: a. K2CO3, DMF, room temperature, 4 hours; b. 4-(2-chloroethyl)morpholine hydrochloride, K2CO3, DMF, 100°C, 4 hours

[0073] Synthesis of compound 9a (2-(4-methoxyphenylamino)-5-hydroxynaphthalene-1,4-dione): 4-methoxyaniline (0.145 gm, 11.7 mmol) and DMF (20 mL) were added to a two-necked RBF (100 mL). K2CO3 (0.217 g, 15.6 mmol) was added to the reaction mixture and stirred at room temperature for 15 minutes. Compound 4 (0.2 gm, 7.84 mmol) was added to the reaction mixture. The reaction mixture was stirred at room temperature for 3 hours. The reaction was monitored by TLC. After completion, the reaction mixture was poured into water (100 mL) and extracted with ethyl acetate (100 mL x 3 times). The organic layer was washed with water (100 mL x 3). The combined organic layer was dried on anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude compound was purified by column chromatography (petroleum ether:ethyl acetate, 70:30). Pure compound = 0.078 gm. Yield = 34%.

[0074] 1 H NMR(CDCl3,400MHz):δ=12.93(s,1H),7.67(d,J=0.8Hz,1H),7.65(m,1H),7.59(d, J=8Hz,1H),7.28(d,J=8.8Hz,2H),6.97(d,J=8.8Hz,2H),6.11(s,1H),3.84(s,3H)

[0075] Synthesis of compound 9b (2-(3,4-dimethoxyphenylamino)-5-hydroxynaphthalene-1,4-dione): 3,4-Dimethoxyaniline (0.72 gm, 47 mmol) and DMF (50 mL) were added to a two-necked RBF (250 mL). K2CO3 (1.08 g, 78.2 mmol) was added to the reaction mixture and stirred at room temperature for 15 minutes. Compound 4 (1.0 gm, 39.2 mmol) was added to the reaction mixture. The reaction mixture was stirred at room temperature for 12 hours. The reaction was monitored by TLC. After completion, the reaction mixture was poured into water (200 mL) and extracted with ethyl acetate (100 mL x 3 times). The organic layer was washed with water (100 mL x 3). The combined organic layer was dried on anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude compound was purified by column chromatography (petroleum ether:ethyl acetate, 80:20). Pure compound = 0.53 gm. Yield = 41%.

[0076] 1 H NMR(CDCl3,400MHz):δ=12.92(s,1H),7.67(d,J=0.8Hz,1H),7.65(m,1H),7.59(d,J=8Hz,1H) ,7.28(d,J=0.8Hz,1H),6.91(d,J=8.8Hz,1H),6.76(d,J=2.4Hz,1H),6.11(s,1H),3.91(s,6H)

[0077] Synthesis of compound formula 37 (5-(2-morpholinoethoxy)-2-(4-methoxyphenylamino)naphthalene-1,4-dione): Compound 9a (0.3 gm, 11.76 mmol) and DMF (30 mL) were added to a two-necked RBF (100 mL). K2CO3 (0.324 g, 23.5 mmol) was added to the reaction mixture and stirred at room temperature for 15 minutes. 4-(2-chloroethyl)morpholine hydrochloride (0.437 gm, 23.5 mmol) was added to the reaction mixture. The reaction mixture was heated at 100 °C for 4 hours. The reaction was monitored by TLC. After completion, the reaction mixture was cooled to room temperature, poured into water (100 mL), and extracted with ethyl acetate (100 mL x 3 times). The organic layer was washed with water (100 mL x 3). The combined organic layer was dried on anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude compound was purified by column chromatography (ethyl acetate:methanol, 95:5). Pure compound = 98 mg. Yield = 21%.

[0078] 1 H NMR(CDCl3,400MHz):δ=12.74(s,1H),7.45(m,2H),7.21(m,3H),7.05(m,2H),5.74 (s,1H),4.06(t,J=6.8Hz,2H),3.91(s,3H),3.64(m,4H),2.69(m,2H),2.49(m,4H)

[0079] Synthesis of compound (5-(2-morpholinoethoxy)-2-(3,4-dimethoxyphenylamino)naphthalene-1,4-dione) of formula 40: Compound 9b (0.3 gm, 9.14 mmol) and DMF (30 mL) were added to a two-necked RBF (100 mL). K2CO3 (0.251 g, 18.3 mmol) was added to the reaction mixture and stirred at room temperature for 15 minutes. 4-(2-chloroethyl)morpholine hydrochloride (0.34 gm, 18.2 mmol) was added to the reaction mixture. The reaction mixture was heated at 100 °C for 4 hours. The reaction was monitored by TLC. After completion, the reaction mixture was cooled to room temperature, poured into water (100 mL), and extracted with ethyl acetate (100 mL x 3 times). The organic layer was washed with water (100 mL x 3). The combined organic layer was dried on anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude compound was purified by column chromatography (ethyl acetate:methanol, 95:5). Pure compound = 35 mg. Yield = 8%.

[0080] 1 H NMR(CDCl3,400MHz):δ=12.74(s,1H),7.48(m,2H),7.21(d,J=2Hz,1H),6.89(d,J=8.4Hz,1H),6.73(m ,2H),5.52(s,1H),4.06(d,J=6.8Hz,2H),3.91(s,6H),3.62(m,4H),2.68(d,J=7.2Hz,2H),2.48(m,4H)

[0081] Scheme 5: [ka] Reagents and conditions: a. Dibromobutane, TBAB, NaOH, H2O, 60°C, 4 hours; b. Morpholine, K2CO3, DMF, room temperature, 12 hours

[0082] Synthesis of compound 11c(5-(4-bromobutoxy)-2-(benzo[d][1,3]dioxol-6-yl)naphthalene-1,4-dione): Dibromobutane (2.2 gm, 103.5 mmol) was added dropwise to a solution of compound 5c (0.35 gm, 10.35 mmol), NaOH (0.082 gm, 20.7 mmol), TBAB (33 mg, 1.35 mmol), and water (30 mL). The reaction mixture was stirred at 60°C for 4 hours. The reaction was monitored by TLC. After the reaction was complete, the reactants were poured into ice-cold water and extracted with ethyl acetate (100 mL x 3 times). The organic layer was washed with water (100 mL). The combined organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude compound was purified by column chromatography (hexane:ethyl acetate, 80:20). Pure compound = 96 mg. Yield = 19%.

[0083] Synthesis of compound 5-(4-morpholinbutoxy)-2-(benzo[d][1,3]dioxol-6-yl)naphthalene-1,4-dione) of formula 2 Morpholine (0.159 gm, 18.2 mmol) was dissolved in DMF (20 mL) and K2CO3 (0.505 gm, 36.4 mmol) at room temperature. Compound 11c (0.087 gm, 18.0 mmol) was added to the reaction mixture at room temperature. The reaction mixture was stirred at room temperature for 16 hours. The reaction was monitored by TLC. After the reaction was complete, the reactants were poured into ice-cold water and extracted with ethyl acetate (100 mL x 3 times). The organic layer was washed with water (100 mL x 3 times). The combined organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude compound was purified by column chromatography (ethyl acetate:methanol, 95:5). Pure compound = 45 mg. Yield = 51%.

[0084] 1 H NMR(CDCl3,400MHz):δ=7.83(d,J=8.0Hz,1H),7.69(d,J=8.2Hz,1H),7.31(d,J=8.2Hz,1H),7.11(m,2H),6.90(m, 2H),6.02(s,2H),4.32(t,J=4.4Hz,2H),3.78(m,4H),3.04(t,J=4.8Hz,2H),2.82(m,4H),1.94(m,2H),1.82(m,2H)

[0085] Scheme 6: [ka] Reagents and conditions: a. Dibromobutane, TBAB, NaOH, H2O, 60°C, 4 hours; b. Morpholine, K2CO3, DMF, room temperature, 12 hours

[0086] Synthesis of compound 13a (5-(4-bromobutoxy)-2-(4-methoxyphenoxy)naphthalene-1,4-dione) Dibromobutane (14.53 gm, 67.5 mmol) was added dropwise to a solution of compound 7a (2.0 gm, 6.75 mmol), NaOH (0.54 gm, 13.5 mmol), TBAB (218 mg, 0.67 mmol), and water (50 mL). The reaction mixture was stirred at 60°C for 3 hours. The reaction was monitored by TLC. After the reaction was complete, the reactants were poured into ice-cold water and extracted with ethyl acetate (100 mL x 4 times). The organic layer was washed with water (100 mL). The combined organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude compound was purified by column chromatography (hexane:ethyl acetate, 80:20). Pure compound = 1.79 gm. Yield = 61%.

[0087] Synthesis of compound 13c(5-(4-bromobutoxy)-2-(benzo[d][1,3]dioxol-5-yloxy)naphthalene-1,4-dione) Dibromobutane (0.5 gm, 16.1 mmol) was added dropwise to a solution of compound 7c (0.35 gm, 10.35 mmol), NaOH (0.129 gm, 32.2 mmol), TBAB (52 mg, 1.61 mmol), and water (20 mL). The reaction mixture was stirred at 60°C for 3 hours. The reaction was monitored by TLC. After the reaction was complete, the reactants were poured into ice-cold water and extracted with ethyl acetate (100 mL x 3 times). The organic layer was washed with water (100 mL). The combined organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude compound was purified by column chromatography (hexane:ethyl acetate, 80:20). Pure compound = 0.4 gm. Yield = 55%.

[0088] Synthesis of the compound (2-(4-methoxyphenoxy)-5-(4-morpholinobtoxy)naphthalene-1,4-dione) of formula 44 Morpholine (0.121 gm, 1.16 mmol) was dissolved in DMF (20 mL) and K2CO3 (0.320 gm, 2.32 mmol) at room temperature. Compound 13a (0.5 gm, 1.16 mmol) was added to the reaction mixture at room temperature. The reaction mixture was stirred at room temperature for 16 hours. The reaction was monitored by TLC. After the reaction was complete, the reactants were poured into ice-cold water and extracted with ethyl acetate (100 mL x 3 times). The organic layer was washed with water (100 mL x 3 times). The combined organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude compound was purified by column chromatography (ethyl acetate:methanol, 95:5). Pure compound = 235 mg. Yield = 48%.

[0089] Synthesis of the compound (5-(4-morpholinbutoxy)-2-(benzo[d][1,3]dioxol-5-yloxy)naphthalene-1,4-dione) of formula 47 Morpholine (0.082 gm, 9.43 mmol) was dissolved in DMF (20 mL) and K2CO3 (0.217 gm, 15.7 mmol) at room temperature. Compound 13c (0.35 gm, 7.86 mmol) was added to the reaction mixture at room temperature. The reaction mixture was stirred at room temperature for 16 hours. The reaction was monitored by TLC. After the reaction was complete, the reactants were poured into ice-cold water and extracted with ethyl acetate (100 mL x 3 times). The organic layer was washed with water (100 mL x 3 times). The combined organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude compound was purified by column chromatography (ethyl acetate:methanol, 95:5). Pure compound = 25 mg. Yield = 7%.

[0090] 1H NMR(CDCl3,400MHz):δ=7.85(d,J=0.8Hz,1H),7.66(dd,J=1.2Hz,8.4Hz,1H),7.32(d,J=0.8Hz,1H),6.83(d,J=8Hz,1H),6.61 (dd,J=2.4Hz,7.6Hz,2H),6.03(s,2H),5.88(s,1H),4.16(t,J=5.6Hz,2H),3.71(m,4H),2.45(m,6H),1.91(m,2H),1.85(m,2H)

[0091] Scheme 7: [ka] Reagents and conditions: a. CuCl, ACN, O2, room temperature, 10 hours; b. Bromine, AcOH, room temperature, 30 minutes

[0092] Synthesis of 5-hydroxy-1,4-naphthoquinone (15): In a 450 mL autoclave reactor, acetonitrile (30 mL) and CuCl (0.78 g, 39.4 mmol) were added gradually at room temperature. A solution of 1,5-dihydroxynaphthalene (1 g, 31.3 mmol) in acetonitrile (200 mL) was added to the reaction mixture at room temperature. 3 kg / cm³ 2 The oxygen pressure was applied to the reaction mixture. The oxygen atmosphere was maintained while vigorously stirring. The reaction mixture was stirred at room temperature for 10 hours. The solution was concentrated under vacuum, and the crude product was purified by column chromatography (hexane:Â=80:20). Pure compound = 0.45 g / m. Yield = 37%. Melting point 157°C; 1 H NMR(300MHz,CDCl3):d=11.91(s,1H),7.69-7.60(m,2H),7.29(dd,J=7.3,2.5Hz,1H),6.96ppm(s,2H)

[0093] Synthesis of 3-bromo-5-hydroxy-(1,4)naphthoquinone (16): 5-Hydroxy-1,4-naphthoquinone (15) (1 g, 57.1 mmol) was suspended in 15 mL of glacial acetic acid. Bromine (1.00 equivalent, 0.3 mL, 57.1 mmol) was added to the reaction mixture at room temperature under light protection. The reaction mixture was stirred under light protection for 20 minutes, then poured onto ice (100 g m). The mixture was stirred vigorously for 30 minutes, and the precipitate was vacuum filtered as an orange solid and washed with a small amount of ice water. The mixture was immediately transferred to a mouthful of RBF and ethanol (8 mL) was added thereto. The reaction mixture was stirred under reflux for 10 minutes using a preheated oil bath. The crude product obtained from the reaction solution as a red solid was vacuum filtered and washed with 5 mL of cold ethanol. The crude product was purified by column chromatography (hexane:HCl, 80:20). Pure compound = 0.7 g m. Yield = 47%.

[0094] Melting point: 168°C; 1 H NMR(300MHz,CDCl3):d=11.73(s,1H),7.68(t,J=7.4Hz,1H),7.64(dd,J=7.4,2.0Hz,1H),7.50(s,1H),7.31ppm(dd,J=7.5,2.0Hz,1H)

[0095] Scheme 8: [ka] Reagents and conditions: a. K2CO3, DMF, room temperature, 4 hours; b. 4-(2-chloroethyl)morpholine hydrochloride, K2CO3, DMF, 100°C, 4 hours

[0096] Synthesis of compound 17a (2-(4-methoxyphenoxy)-8-hydroxynaphthalene-1,4-dione) 4-methoxyphenol (0.972 gm, 78.4 mmol) and DMF (50 mL) were added to a two-necked RBF (250 mL). K2CO3 (1.08 g, 78.4 mmol) was added to the reaction mixture and stirred at room temperature for 15 minutes. Compound 16 (2.0 gm, 78.4 mmol) was added to the reaction mixture. The reaction mixture was stirred at room temperature for 3 hours. The reaction was monitored by TLC. After completion, the reaction mixture was poured into water (200 mL) and extracted with ethyl acetate (100 mL x 3 times). The organic layer was washed with water (100 mL x 3). The combined organic layer was dried on anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude compound was purified by column chromatography (petroleum ether:ethyl acetate, 95:5). Pure compound = 1.0 gm. Yield = 43%.

[0097] Synthesis of compound 17b (2-(4-fluorophenoxy)-8-hydroxynaphthalene-1,4-dione) 4-fluorophenol (0.879 gm, 78.4 mmol) and DMF (50 mL) were added to a two-necked RBF (100 mL). K2CO3 (1.08 g, 78.4 mmol) was added to the reaction mixture and stirred at room temperature for 15 minutes. Compound 16 (2.0 gm, 78.4 mmol) was added to the reaction mixture. The reaction mixture was stirred at room temperature for 3 hours. The reaction was monitored by TLC. After completion, the reaction mixture was poured into water (200 mL) and extracted with ethyl acetate (100 mL x 3 times). The organic layer was washed with water (100 mL x 3). The combined organic layer was dried on anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude compound was purified by column chromatography (petroleum ether:ethyl acetate, 95:5). Pure compound = 1.13 gm. Yield = 51%.

[0098] Synthesis of compound 17c (2-(benzo[d][1,3]dioxol-6-yloxy)-8-hydroxynaphthalene-1,4-dione) Sesamol (1.08 gm, 78.4 mmol) and DMF (50 mL) were added to a two-necked RBF (100 mL). K2CO3 (0.54 g, 78.4 mmol) was added to the reaction mixture and stirred at room temperature for 15 minutes. Compound 4 (2.0 gm, 78.4 mmol) was added to the reaction mixture. The reaction mixture was stirred at room temperature for 4 hours. The reaction was monitored by TLC. After completion, the reaction mixture was poured into water (200 mL) and extracted with ethyl acetate (100 mL x 3 times). The organic layer was washed with water (100 mL x 3). The combined organic layer was dried on anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude compound was purified by column chromatography (petroleum ether:ethyl acetate, 95:5). Pure compound = 0.676 gm. Yield = 25%.

[0099] Synthesis of the compound (8-(2-morpholinoethoxy)-2-(4-fluorophenoxy)naphthalene-1,4-dione) of formula 80 Compound 17b (0.5 gm, 17.66 mmol) and DMF (20 mL) were added to a two-necked RBF (100 mL). K2CO3 (0.488 g, 35.33 mmol) was added to the reaction mixture and stirred at room temperature for 15 minutes. 4-(2-chloroethyl)morpholine hydrochloride (0.394 gm, 21.2 mmol) was added to the reaction mixture. The reaction mixture was heated at 100 °C for 3 hours. The reaction was monitored by TLC. After completion, the reaction mixture was cooled to room temperature, poured into water (100 mL), and extracted with ethyl acetate (100 mL x 3 times). The organic layer was washed with water (100 mL x 3). The combined organic layer was dried on anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude compound was purified by column chromatography (ethyl acetate:methanol, 95:5). Pure compound = 35 mg. Yield = 33%.

[0100] 1H NMR(CDCl3,400MHz):δ=7.71(d,J=1.2Hz,1H),7.68(m,1H),7.31(dd,J=1.2Hz & 7.6Hz,1H),7.17(m,4H),5.84(s,1H),4.31(t,J=5.6Hz,2H),3.76(m,4H),2.98(t,J=5.6Hz,2H),2.72(m,4H)

[0101] Synthesis of the compound (8-(2-morpholinoethoxy)-2-(benzo[d][1,3]dioxol-6-yloxy)naphthalene-1,4-dione) of formula 79 Compound 7c (0.5 gm, 17.66 mmol) and DMF (20 mL) were added to a two-necked RBF (100 mL). K2CO3 (0.49 g, 35.3 mmol) was added to the reaction mixture and stirred at room temperature for 15 minutes. 4-(2-chloroethyl)morpholine hydrochloride (0.395 gm, 21.2 mmol) was added to the reaction mixture. The reaction mixture was heated at 100 °C for 2 hours. The reaction was monitored by TLC. After completion, the reaction mixture was cooled to room temperature, poured into water (100 mL), and extracted with ethyl acetate (100 mL x 3 times). The organic layer was washed with water (100 mL x 3). The combined organic layer was dried on anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude compound was purified by column chromatography (ethyl acetate:methanol, 95:5). Pure compound = 294 mg. Yield = 42%.

[0102] 1 H NMR (CDCl3,400MHz): δ=7.74(d,J=0.8Hz,1H),7.68(dd,J=1.2Hz,8.4Hz,1H),7.29(d,J=0.8Hz,1H),6.83(d,J=8Hz,1H),6.60(dd,J=2. 4Hz,7.6Hz,1H),6.56(d,J=2.4Hz,1H),6.03(s,2H),5.91(s,1H),4.30(t,J=5.6Hz,2H),3.76(m,4H),2.98(t,J=5.6Hz,2H),2.72(m,4H)

[0103] Test data The following tests were conducted to determine the efficacy and non-toxicity of the compound.

[0104] Cancer cell assay 1. In vitro antiproliferative assay (MTT assay) The MTT assay is a simple and highly sensitive assay for measuring the metabolic reduction activity of cells. The increase in this activity over time is interpreted as a parameter of cell growth. If drug treatment reduces this increase, the effect is growth inhibition, cell toxicity, or both. The compounds of the present invention and standard cytotoxic agents (e.g., cisplatin) were tested at different concentrations (1, 0.1, 0.01, 0.001 mM) using breast and prostate cancer cell lines. All cell lines were cultured in a 37°C incubator under a 5% CO2 environment. The compounds were dissolved in DMSO at a concentration of 0.1 M (stock solution). Cells were seeded in 96-well plates with appropriate seeding efficiency.

[0105] The following seeding efficiencies were standardized against the MTT assay.

[0106] [Table 1]

[0107] In the MTT procedure, cells were seeded into a 96-well plate according to the specified seeding efficiency (Table 1). The plate was then incubated at 37°C for 24 hours in a 5% CO2 atmosphere. Next, the appropriate concentration of the drug was added to the plate, and a further incubation was performed for 48 hours (37°C in a 5% CO2 atmosphere). The assay plate was then centrifuged twice at 3000 rpm for 3 minutes, followed by discarding the supernatant. Next, 100 μL of MTT solution (0.5 mg / mL) was added to each well of the plate, and it was incubated for a further 4 hours (37°C in a 5% CO2 atmosphere). After 4 hours of incubation, the plate was centrifuged twice, and the supernatant was carefully aspirated and removed. Next, 200 μL of DMSO was added to each well and dissolved. The MTT crystals were thoroughly mixed by shaking the plate. Subsequently, a logarithmic XY graph of viability was plotted against the logarithmic drug concentration. Next, the IC50 (the drug concentration that inhibits 50% of the cell population) was calculated using regression analysis.

[0108] Results of MTT assays of compounds against breast cancer (MDAMB231 cell line) and prostate cancer (PC3 cell line) [Table 2]

[0109] The table above shows that the compound exhibits significantly higher efficacy against breast cancer and prostate cancer cell lines in the MTT assay compared to the standard therapeutic agent, cisplatin.

[0110] Figures 1 to 50 show the activity of compounds with formulas 1, 2, 7, 37, 40, 41, 43, 46, 47, 52, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, and 81 against breast cancer cell lines and prostate cancer cell lines, respectively, compared to cisplatin. These compounds were found to exhibit higher anticancer activity compared to cisplatin.

[0111] 2. Soft agar assay The soft agar colony formation assay is a scaffold-independent growth assay in soft agar and is one of the most stringent assays for detecting malignant transformation of cells. For this assay, malignant cells are cultured in soft agar medium for 1-2 weeks with a suitable control. After this incubation period, the formed colonies can be morphologically analyzed by either cell staining or quantification of the number of colonies formed. The assay results are comparable to those obtained after injecting tumorigenic cells into nude mice and are considered the "optimal criterion" for testing the oncogenicity of cells (one of the key features of cancer stem cells (CSCs)) in vitro.

[0112] In short, for the soft agar assay, a mixture of 50 μL of 2× medium (appropriately used depending on the cell line) and 50 μL of 1.2% Bacto Agar was placed in each well of a 96-well microtiter assay plate. 10 μL of cells (cells with a specific seeding efficiency pre-standardized for each cell line) were mixed in a vial with 20 μL of 2× medium, 30 μL of 0.8% Bacto Agar, and 1.6 μL of drug (drug at an appropriate concentration) and transferred to the solidified pre-layers of the assay plate. The cells were then grown at 37°C and 5% CO2 for one week to form colonies. Three days after setting up the experiment, 50 μL of appropriate 2× medium was intermittently supplied. Subsequently, 16 μL of Alamar Blue (1.5 mg / mL) was added to all wells, and the resulting colonies were quantified. The plate was incubated at 37°C for 24 hours. Absorbance was then measured at 630 nm. Next, the logarithmic XY graph of survival rates was plotted against the logarithmic scale of drug concentrations. Then, the IC50 (the drug concentration that inhibits 50% of the cell population) was calculated using regression analysis.

[0113] The following seeding efficiencies were standardized against the soft agar assay.

[0114] [Table 3]

[0115] Results of soft agar assays of compounds against breast cancer (MDAMB231 cell line) and prostate cancer (PC3 cell line) [Table 4]

[0116] The table above shows that the compound exhibits significantly higher efficacy against breast cancer and prostate cancer cell lines in soft agar assays compared to the standard therapeutic agent, cisplatin.

[0117] Figure 51 shows the activity of compounds with formulas 2, 40, 41, 43, 52, 67, 68, 71, 72, and 73 against breast cancer cell lines, compared to cisplatin. These compounds were found to exhibit higher anticancer activity compared to cisplatin.

[0118] Figure 52 shows the activity of compounds with formulas 2, 40, 41, 43, 52, 67, 68, 71, 72, and 73 against prostate cancer cell lines, compared to cisplatin. These compounds were found to exhibit higher anticancer activity compared to cisplatin.

[0119] 3. Stem cell assay In vitro sphere formation assay: The sphere assay measures the ability of cancer stem cells (CSCs) to form spheres in a specially designed serum-free medium. Using this assay, the killing efficiency of the test compound was measured compared to that of the standard chemotherapy drug cisplatin.

[0120] Materials and Reagents: 50×B27 Supplement (Life Technologies, Invitrogen, Catalog No.: 17502-044), Fibroblast Growth Factor (FGF) (Sigma-Aldrich, Catalog No.: F029125), Epidermal Growth Factor (EGF) (Sigma-Aldrich, Catalog No.: E9644), Insulin (Sigma, Catalog No.: 19278), Dulbecco's Modified Eagle Medium / F12 (HiMedia, Catalog No.: AL139-6), Dulbecco's Phosphate-Buffered Saline (HiMedia, Catalog No.: TL1006), Trypan Blue (TC193), Prostatic Epithelial Cell Medium (LONZA, Catalog No.: CC-3166), MEGM (LONZA, Catalog No.: CC-3051), Heparin (Sigma, Catalog No.: H3393), Penstrep (HiMedia, Catalog No.: A002)

[0121] Preparation of mammothsphere medium (100 mL): 1 g of methylcellulose was placed in a magnetic stirrer and autoclaved. While stirring with the magnetic stirrer, 100 mL of simple medium (MEBM) was added and dissolved. After complete dissolution, 80 μL of FGF, 40 μL of EGF, 1 mL of Penstrep, and 400 μL of heparin were added.

[0122] Preparation of Prostosphere medium (100 mL): 1 g of methylcellulose was placed in a magnetic stirrer and autoclaved. While stirring with the magnetic stirrer, 100 mL of simple medium (basic medium for prostatic epithelial cells) was added and dissolved. After complete dissolution, 40 μL of insulin, 2 mL of B27, 80 μL of EGF, and 1 mL of Penstrep were added.

[0123] Procedure - Cells were trypsinized and passed through cell strainers (100 μl and 40 μl, respectively) to obtain single-cell suspensions. Cells were diluted to a concentration of 2000 cells / 100 μL and suspended in either mammosphere (for breast cancer cell lines) or prostosphere (for prostate cancer cell lines). 100 μL of this suspension was added to each well of a 96-well suspension plate and incubated at 37°C, 5% CO2 for 24 hours. An appropriate concentration of drug (2 μL) was added to each well containing 100 μL of stem cell culture medium. The plate was incubated at 37°C, 5% CO2 for 72 hours. After incubation, 2.5 μL of the respective concentration of drug and 50 μL of stem cell culture medium were added to each well, and the plate was incubated for a further 72 hours at 37°C, 5% CO2. After incubation, 3 μL of each drug concentration was added again with 50 μL of stem cell culture medium, and the plates were re-incubated at 37°C and 5% CO2 for 72 hours. The number of primary spheres formed at each concentration was counted. The spheres were converted to sphere viability compared to untreated cells (Growth Control with DMSO, GCD). The sphere viability comparison graph was plotted against drug concentration and compared to cisplatin, the standard therapeutic agent.

[0124] Results of in vitro sphere formation assays of compounds against breast cancer (MDAMB231 cell line) and prostate cancer (PC3 cell line) at a seeding efficiency of 2000 cells / well (n=6+SD). [Table 5-1]

[0125] [Table 5-2]

[0126] [Table 5-3]

[0127] The above results indicate that the above compound is more effective than cisplatin in inhibiting the MDAMB231 sphere.

[0128] Figures 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, and 49 relate to the compounds of formulas 1, 2, 7, 37, 40, 41, 43, 46, 47, 52, and 67-81, respectively, in the MDAMB231 cell line.

[0129] Figures 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, and 49 show the sphere viability obtained by converting the number of spheres formed and compared with a growth control (GCD) using DMSO. The GCD is considered to have a 100% viability rate. The sphere count results at each drug concentration shown in Table 6 were converted to sphere viability for graphical display. Figures and Table 6 show that the sphere viability of MDAMB231 is reduced compared to cisplatin in the presence of compounds of formulas 1, 2, 7, 37, 40, 41, 43, 46, 47, 52, and 67-81.

[0130] [Table 6-1]

[0131] [Table 6-2]

[0132] [Table 7-1]

[0133] [Table 7-2]

[0134] [Table 7-3]

[0135] The above results indicate that the compounds of formulas 1, 2, 7, 37, 40, 41, 43, 46, 47, 52, and 67-81 are more effective than cisplatin in inhibiting the PC3 sphere.

[0136] Figures 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, and 50 relate to compounds of formulas 1, 2, 7, 37, 40, 41, 43, 46, 47, 52, and 67-81, respectively.

[0137] Figures 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, and 50 show sphere viability obtained by converting the number of spheres formed and compared with a growth control (GCD) using DMSO. GCD is considered to be 100% viability. The sphere count results at each drug concentration shown in Table 8 were converted to sphere viability for graphical display. Figures and Table 8 show that the sphere viability of PC3 is reduced compared to cisplatin in the presence of compounds of formulas 2, 7, 37, 40, 41, 43, 46, 47, 52, and 67-81.

[0138] [Table 8]

[0139] [Table 8-2]

[0140] 4. Activity against lymphocytes Lymphocyte assay Human lymphocytes were isolated from peripheral blood. A pure lymphocyte population was obtained by fractionation centrifugation in which diluted defibrinated blood was layered onto a solution of sodium diatrizoate and polysucrose (HiSep LSM 1077) and centrifuged at low speed for 30 minutes.

[0141] Procedure: Separation of lymphocytes from fresh defibrinated blood was performed by the procedure described below. 1. Diluted fresh defibrinated blood was layered little by little onto (HiSeP LSM1077) and centrifuged at low speed for 30 minutes. 2. The lymphocyte layer (buffy coat) was carefully taken out into a new collection tube. 3. The buffy coat was further washed with Dulbecco's phosphate buffered saline (D.P.B.S) of dilution buffer. 4. The supernatant was discarded and the pellet was resuspended in D.P.B.S. 5. The viability of the cells was confirmed with a hemocytometer. 6. Cells with purity and viability exceeding 95% were collected for experiments. 7. The purified lymphocytes were diluted to a concentration of 700,000 cells / mL with sterile D.P.B.S, and the MTT method was carried out strictly as described above.

[0142]

Table 9

[0143] The above table and Figure 5-3 show that the activity of the compounds is high against cancer cells compared to normal cells, indicating the safety of these compounds.

[0144] 5. Wound healing effect Wound Healing Assay (WHA): The Wound Healing Assay (WHA) evaluates the ability of cancer stem cells to heal wounds formed in a confluent monolayer. Using this assay, the ability of the test drug to inhibit the wound healing ability of cancer stem cells was measured compared to that of standard chemotherapy drugs such as cisplatin.

[0145] Procedure: 0.35 × 10 6 Cells were seeded into each well of a 6-well tissue culture plate. The plates were incubated at 37°C and 5% CO2 for 48 hours. After observing that the cells were fully confluent and washing twice with DPBS, the center of each well was scratched laterally using a sterile 100 μl tip. The width of the scratch was measured at 0 hours immediately after scratching. Each compound at its IC10 concentration was added to each well. The plates were incubated at 37°C and 5% CO2, and the width of the scratch was measured at various time intervals, such as 6 hours, 24 hours, and 48 hours. The average of three distances was taken for each time point using IS camera Measure. The average width of the scratch in micrometers was plotted against the time interval after treatment. Anti-CSC capacity was identified by calculating the inhibition rate of each compound after 48 hours compared to cisplatin.

[0146] [Table 10]

[0147] The table and Figure 54 above show that anticancer compounds inhibit wound healing in cancer cells, thereby preventing cancer spread compared to the standard treatment drug cisplatin.

[0148] 6. Inhibitory effect of compounds on aldehyde dehydrogenase (ALDH), a cancer marker. Aldehyde dehydrogenase (ALDH) assay: Aldehyde dehydrogenases (ALDHs) are a family of enzymes that catalyze the metabolism of exogenous and endogenous aldehydes and prevent the accumulation of aldehydes and their metabolites, which can be reactive and toxic. In addition to their role in aldehyde metabolism, ALDH enzymes also play important roles in other cellular processes such as cell proliferation, differentiation, and survival.

[0149] ALDH also functions as a marker for specific stem cell populations, including hematopoietic stem cells and certain cancer stem cells.

[0150] ALDH concentration was determined using the (Kinesis Dx) ELISA kit according to the following protocol. 1. Standard (50 μl) / sample (40 μl) was added to each well (except for the blank). 2. Next, biotin-labeled antibody (10 μl) was added to each sample well (except for the blank). 3. Next, horseradish peroxidase (HRP) labeled solution (50 μl) was added to each well (except the blank). 4. The plates were incubated in an incubator at 37°C for 1 hour. 5. Wash the plate (four times with washing buffer), and lightly tap the plate on absorbent paper to aspirate any remaining buffer. Since any residue may interfere with the reading process, all liquid should be cleared from the bottom of the microtiter well. 6. Add TMB substrate A (50 μl) to each well, including the blank well, followed by TMB substrate B (50 μl). 7. Incubate in a dark place at 37°C for 10 minutes. 8. Add stop solution (50 μl). The wells should change from blue to yellow. Read the absorbance at 9.450 nm. 10. An XY graph of concentration versus optical density was plotted. The ALDH concentration was calculated by substituting the optical density values ​​into the regression analysis equation.

[0151] [Table 11]

[0152] The table and Figure 55 above show that the compound inhibits aldehyde dehydrogenase (ALDH), a cancer stem cell (CSC) marker, compared to the standard therapeutic agent cisplatin.

Claims

1. A compound represented by any one of the following structural formulas. 【Chemistry 1】 【Chemistry 2】 【Transformation 3】 【Chemistry 4】

2. A pharmaceutical composition comprising the compound described in Claim 1 and at least one pharmaceutically acceptable additive, in the presence or absence of one or more active ingredients.

3. The compound according to claim 1, for use in the treatment of uncontrolled cell growth or cancer.

4. The compound according to claim 1, for use in combination with at least one standard treatment for cancer to treat uncontrolled cell growth or cancer.

5. A compound for treating or inhibiting uncontrolled cell growth or cancer by administering an effective amount of the compound to a patient, wherein the cancer is breast cancer, prostate cancer, brain cancer, hematological cancer, bone marrow cancer, liver cancer, pancreatic cancer, skin cancer, kidney cancer, colon cancer, ovarian cancer, lung cancer, testicular cancer, penile cancer, thyroid cancer, parathyroid cancer, pituitary cancer, thymic cancer, retinal cancer, uveal cancer, conjunctival cancer, spleen cancer, head cancer, neck cancer, tracheal cancer, gallbladder cancer, rectal cancer, salivary gland cancer, adrenal gland cancer, throat cancer, esophageal cancer, lymph node cancer, sweat gland cancer, sebaceous gland cancer, muscle cancer, heart cancer, and stomach cancer, according to claim 1.

6. A pharmaceutical composition for treating or inhibiting uncontrolled cell growth or cancer by administering an effective amount of the pharmaceutical composition to a patient, wherein the cancer is breast cancer, prostate cancer, brain cancer, hematological cancer, bone marrow cancer, liver cancer, pancreatic cancer, skin cancer, kidney cancer, colon cancer, ovarian cancer, lung cancer, testicular cancer, penile cancer, thyroid cancer, parathyroid cancer, pituitary cancer, thymic cancer, retinal cancer, uveal cancer, conjunctival cancer, spleen cancer, head cancer, neck cancer, tracheal cancer, gallbladder cancer, rectal cancer, salivary gland cancer, adrenal gland cancer, throat cancer, esophageal cancer, lymph node cancer, sweat gland cancer, sebaceous gland cancer, muscle cancer, heart cancer, and stomach cancer, according to claim 2.

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