Compound, pharmaceutical composition, kit and application thereof

Abstract

The invention relates to a compound, a pharmaceutical composition, a kit and application thereof. The present invention relates to a compound contained within formula VII, IX, XI or XIII, R1 being selected from the group consisting of hydrogen or optionally substituted alkyl or halogen; r2 is selected from the group consisting of an alkyl group, an optionally substituted C4-C8 cycloalkyl group, an optionally substituted C4-C8 heteroalkyl group, an optionally substituted C4-C8 heterocycloalkyl group, an optionally substituted C5-C6 aryl group, and an optionally substituted C4-C6 heteroaryl group; r3a, R3b, R3c or R3d are selected from hydrogen, halogen, methyl, methoxyl, trifluoromethyl, hydroxyl or cyano; r4 is selected from alkyl or heteroalkyl; a comprises a cycloalkyl group, a heterocycloalkyl group, an aryl group or a heteroaryl group; x is selected from hydrogen, halogen, amino, heterocyclic alkyl or hydroxyl; y is an O atom, an S atom or an N atom; m is an integer selected from 0 to 9. The invention also relates to a method for preparing the compound and a use thereof. # imgabs0 #

Description

[0001] This application is a divisional application of the invention patent application with application number 202080026341.X, filed on March 6, 2020, and entitled “Substituted Bicyclic and Tetracyclic Quinones and Related Methods of Use”. Statement Regarding Federally Sponsored Research or Development

[0002] This invention was made with government support under Grant CA188252 from the National Institutes of Health. The government has certain rights in this invention. Technical Field

[0003] The present invention belongs to the field of medicinal chemistry. In particular, the present invention relates to small molecule compounds having bicyclic and tetracyclic quinone scaffolds, which have antiproliferative activity by, for example, inducing reactive oxygen species. The present invention further relates to methods for preparing these compounds and methods for using these compounds to treat cancer (e.g., pancreatic cancer, leukemia, non-small cell lung cancer, colon cancer, CNS cancer, melanoma, ovarian cancer, breast cancer, kidney cancer and prostate cancer). Background Art

[0004] Cancer is the second leading cause of death in Europe and North America. Surgery, radiation, chemotherapy and immunotherapy are the main methods for treating various cancers. Despite significant progress over the past decade, the cure rate for most cancers remains low. Therefore, novel therapies are urgently needed to improve the survival rate of patients with these devastating diseases. Cancer cells accumulate a large number of mutations during their evolutionary process, changing multiple signal transduction pathways and networks. Therefore, compared with single-agent drugs, a combination of multiple drugs or single compounds with multiple targets tends to show excellent activity. Among the various types of small molecule drugs tested, quinone-containing compounds show great prospects because they significantly increase the oxygen consumption rate of treated cells. Quinones can inhibit various pathways due to their redox, metal chelation, and in some cases, the reactivity of nucleophiles by Michael addition. More than two dozen drugs containing quinone moieties have been approved by the FDA or are undergoing clinical studies, not only in oncology but also for other diseases. For example, doxorubicin and its dozens of analogs mitoxantrone and mitomycin C are some of the most commonly used FDA-approved chemotherapeutic agents for a variety of cancers containing quinone groups.

[0005] There is a need for improved small molecule compounds having bicyclic and tetracyclic quinone scaffolds for the treatment of cancer.

[0006] The present invention addresses this need. Summary of the invention

[0007] Altered redox homeostasis in cancer cells provides new opportunities for tumor intervention. Reactive oxygen species (ROS), which are natural byproducts of mitochondrial respiration, play important roles as second messengers in cell signaling. In tumor cells, antioxidant enzymes are often active as a result of elevated intrinsic ROS levels. Altered redox homeostasis in tumors makes them more susceptible to induced oxidative stress, which overwhelms their adaptive antioxidant capacity and triggers ROS-mediated cell death.

[0008] Experiments conducted during the development process synthesized a variety of new small molecules with bicyclic and tetracyclic quinone scaffolds that have improved metabolic stability, solubility, and pharmacokinetic properties. Indeed, as described in Example III, various compounds of the present invention were shown to inhibit the growth of various cancer cell lines (see Tables 2, 3, 4, and 5; Figure 1 , 2 , 3, 4 and 5).

[0009] Thus, the present invention provides a new class of small molecules having bicyclic and tetracyclic quinone structures with improved metabolic stability, solubility and pharmacokinetic properties. The present invention further provides the use of such small molecules as therapeutic agents for the treatment of cancer.

[0010] Thus, the present invention contemplates that exposing an animal (e.g., a human) having cancer (e.g., and / or a cancer-related disorder) to a therapeutically effective amount of one or more agents having bicyclic and tetracyclic quinone structures will completely inhibit the growth of cancer cells or supporting cells and / or render such cells as a population more susceptible to the cell death-inducing activity of the cancer therapeutic drug or radiation therapy.

[0011] The present invention anticipates that the compounds of the present invention (e.g., compounds having bicyclic and tetracyclic quinone structures) will fill an unmet need for the treatment of a variety of cancer types when administered as a monotherapy to induce cell growth inhibition, apoptosis and / or cell cycle arrest in cancer cells, or when administered in a timed relationship with one or more additional agents (such as other cell death inducing or cell cycle disrupting cancer therapeutic drugs) or radiation therapy (combination therapy) to render a greater proportion of cancer cells or supporting cells susceptible to apoptotic programming compared to a corresponding proportion of cells in an animal treated with the cancer therapeutic drug alone or radiation therapy alone.

[0012] In certain embodiments of the invention, animals treated with a therapeutically effective amount of a compound of the invention in combination with an anticancer agent course of treatment produce greater tumor response and clinical benefit in such animals than animals treated with the compound or anticancer drug / radiation alone. Since the dosages for all approved anticancer drugs and radiation therapy are known, the present invention contemplates various combinations of them with the compounds of the invention.

[0013] Certain compounds of the invention having an indole scaffold may exist as stereoisomers including optical isomers. The invention includes all stereoisomers, both as pure individual stereoisomer preparations and preparations enriched for each stereoisomer, as well as racemic mixtures of such stereoisomers and both individual diastereomers and enantiomers that can be separated according to methods well known to those skilled in the art.

[0014] In a specific embodiment, compounds having bicyclic and tetracyclic quinone structures (or analogs) contained within Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV, XVI and XVII are provided:

[0015]

[0016]

[0017] This includes pharmaceutically acceptable salts, solvates and / or prodrugs thereof.

[0018] In some embodiments, R 1 Selected from the group consisting of hydrogen, halogen, alkyl, heteroalkyl, optionally substituted C6-C 14 aryl and optionally substituted 5- to 14-membered heteroaryl.

[0019] In some embodiments, R 1 Selected from the group consisting of: H,

[0020] In some embodiments, R 2 Selected from the group consisting of alkyl, optionally substituted C4-C8 cycloalkyl, optionally substituted C4-C8 heteroalkyl, optionally substituted C4-C8 heterocycloalkyl, optionally substituted C5-C6 aryl and optionally substituted C4-C6 heteroaryl.

[0021] In some embodiments, R 2 Optionally selected from the group consisting of:

[0022]

[0023] In some embodiments, R 3a , R 3b , R 3c or R 3d Selected from the group consisting of hydrogen, halogen, methyl, methoxy, trifluoromethyl, hydroxyl or cyano.

[0024] In some embodiments, R 3a, R 3b , R 3c or R 3d is optionally mono- or poly-substituted and selected from the group consisting of:

[0025]

[0026] In some embodiments, R 4 Selected from the group consisting of alkyl or heteroalkyl.

[0027] In some embodiments, R 4 Optionally selected from the group consisting of:

[0028]

[0029] In some embodiments, A includes cycloalkyl, heterocycloalkyl, aryl, or heteroaryl.

[0030] In some embodiments, X is selected from the group consisting of hydrogen, halogen, amino, heterocycloalkyl, or hydroxy.

[0031] In some embodiments, X is optionally selected from the group consisting of:

[0032]

[0033] In some embodiments, n is an integer selected from 0 to 5.

[0034] In some embodiments, m is an integer selected from 0 to 9.

[0035] In some embodiments, Y is independently selected from O, S, and N atoms.

[0036] In some embodiments, W is independently selected from C atoms, N atoms.

[0037] In some embodiments, for Formulas I-XVII, the compounds shown in Table 1 are contemplated.

[0038] Table 1. Structures of representative compounds expected for Formulas I-XVII.

[0039]

[0040]

[0041]

[0042]

[0043]

[0044]

[0045]

[0046]

[0047]

[0048]

[0049]

[0050] In some embodiments, any of the compounds described herein is a 2,2,2-trifluoroacetate (TFA) salt.

[0051] The invention further provides methods for preparing any of the compounds of the invention by following at least some of the techniques listed in the Experimental Section.

[0052] The present invention also provides the use of compounds for inducing cell cycle arrest and / or apoptosis in cancer cells. The present invention also relates to the use of compounds for sensitizing cells to one or more additional agents (such as inducers of apoptosis and / or cell cycle arrest) and for chemoprotecting normal cells by inducing cell cycle arrest prior to treatment with chemotherapeutic agents.

[0053] The compounds of the present invention can be used to treat, improve or prevent diseases, such as those diseases that respond to induction of apoptotic cell death, for example, diseases characterized by dysregulation of apoptosis, including hyperproliferative diseases, such as cancer. In some embodiments, the compound can be used to treat, improve or prevent cancers characterized by resistance to cancer therapy (for example, those cancer cells with chemical resistance, radioactivity resistance, hormone resistance, etc.). In some embodiments, cancer is multiple myeloma, acute myeloid leukemia, melanoma, breast cancer, head or neck cancer, colon cancer, lung cancer, ovarian cancer, prostate cancer and / or pancreatic cancer.

[0054] The present invention also provides pharmaceutical compositions comprising a compound of the present invention in a pharmaceutically acceptable carrier.

[0055] The invention also provides a kit comprising a compound of the invention and instructions for administering the compound to an animal. The kit may optionally contain other therapeutic agents, such as anti-cancer agents or apoptosis modulators. BRIEF DESCRIPTION OF THE DRAWINGS

[0056] Figure 1 : Growth inhibition of compound 1 in NCI60 cell line (% control).

[0057] Figure 2 : Growth inhibition of compound 13 in NCI60 cell line (% control).

[0058] Figure 3 : Growth inhibition of compound 49 in NCI60 cell line (% control).

[0059] Figure 4 : Growth inhibition of compound 50 in NCI60 cell line (% control).

[0060] Figure 5 : Cytotoxicity of compound 13 in NCI60 cell line. DETAILED DESCRIPTION

[0061] definition

[0062] As used herein, the term "anti-cancer agent" refers to any therapeutic agent (e.g., a chemotherapeutic compound and / or a molecular therapeutic compound), antisense therapy, radiation therapy, or surgical intervention used to treat a hyperproliferative disease, such as cancer (e.g., in a mammal, e.g., in a human).

[0063] The term "prodrug" as used herein refers to a pharmacologically inactive derivative of the parent "drug" molecule that needs to be biotransformed (e.g., spontaneously or enzymatically) in the target physiological system to release the active drug or convert the prodrug (e.g., enzymatically, physiologically, mechanically, electromagnetically) to the active drug. Prodrugs are intended to overcome problems associated with stability, water solubility, toxicity, lack of specificity or limited bioavailability. Exemplary prodrugs include the active drug molecule itself and a chemical masking group (e.g., a group that reversibly inhibits the activity of the drug). Some prodrugs are modifications or derivatives of compounds with groups that are cleavable under metabolic conditions. Prodrugs can be readily prepared from the parent compound using methods known in the art, such as those described in: A Textbook of Drug Design and Development, Krogsgaard-Larsen and H. Bundgaard (eds.), Gordon & Breach, 1991, especially Chapter 5: "Design and Applications of Prodrugs"; Design of Prodrugs, H. Bundgaard (ed.), Elsevier, 1985; Prodrugs: Topical and Ocular Drug Delivery, KB Sloan (ed.), Marcel Dekker, 1998; Methods in Enzymology, K. Widder et al. (eds.), Vol. 42, Academic Press, 1985, especially pp. 309-396; Burger's Medicinal Chemistry and Drug Discovery, 5th Ed., M. Wolff (ed.), John Wiley & Sons, 1995, especially Vol. 1 and pp. 172-178 and pp. 949-982; Pro-Drugs as Novel Delivery Systems, T. Higuchi and V. Stella (eds.), Am. Chem. Soc., 1975; and Bioreversible Carriers in Drug Design, EB Roche (ed.), Elsevier, 1987.

[0064] When exemplary prodrugs undergo solvolysis or enzymatic degradation or other biochemical transformations (e.g., phosphorylation, hydrogenation, dehydrogenation, glycosylation) under physiological conditions, they become pharmaceutically active in vivo or in vitro. Prodrugs generally provide advantages of water solubility, tissue compatibility, or delayed release in mammalian organisms. (See, e.g., Bundgard, Design of Prodrugs, pp. 7-9, 21-24, Elsevier, Amsterdam (1985); and Silverman, The Organic Chemistry of Drug Design and Drug Action, pp. 352-401, Academic Press, San Diego, CA (1992)). Common prodrugs include acid derivatives, such as esters prepared by reacting the parent acid with a suitable alcohol (e.g., a lower alkanol) or esters prepared by reacting the parent alcohol with a suitable carboxylic acid (e.g., an amino acid), amides prepared by reacting the parent acid compound with an amine, basic groups that are reacted to form acylated base derivatives (e.g., lower alkyl amides), or phosphorus-containing derivatives, such as phosphates, phosphonates, and phosphoramidates, including cyclic phosphates, phosphonates, and phosphoramidates (see, e.g., U.S. Patent Application Publication No. US 2007 / 0249564A1; incorporated herein by reference in its entirety).

[0065] As used herein, the term "pharmaceutically acceptable salt" refers to any salt of the compound of the present invention that is physiologically tolerated in a target animal (e.g., a mammal) (e.g., obtained by reacting with an acid or base). The salt of the compound of the present invention may be derived from an inorganic or organic acid and base. Examples of acids include, but are not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, perchloric acid, fumaric acid, maleic acid, phosphoric acid, glycolic acid, lactic acid, salicylic acid, succinic acid, toluene-p-sulfonic acid, tartaric acid, acetic acid, citric acid, methanesulfonic acid, ethanesulfonic acid, formic acid, benzoic acid, malonic acid, sulfonic acid, naphthalene-2-sulfonic acid, benzenesulfonic acid, etc. In obtaining the compounds of the present invention and their pharmaceutically acceptable acid addition salts, other acids, such as oxalic acid, although they are not pharmaceutically acceptable themselves, can be used as intermediates in the preparation of salts.

[0066] Examples of bases include, but are not limited to, alkali metal (eg, sodium) hydroxides, alkaline earth metal (eg, magnesium) hydroxides, ammonia, and compounds of formula NW4 + The compound wherein W is C 1-4 Alkyl, etc.

[0067] Examples of salts include, but are not limited to, acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, flucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, chloride, bromide, iodide, 2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, palmoate, pectinate, persulfate, phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, toluenesulfonate, undecanoate, and the like. Other examples of salts include pyrophosphate, ... + NH4 + and NW4 + (W is C 1-4 For therapeutic use, salts of the compounds of the present invention are considered to be pharmaceutically acceptable. However, it is also found that salts of non-pharmaceutically acceptable acids and bases can be used, for example, in the preparation or purification of pharmaceutically acceptable compounds.

[0068] As used herein, the term "solvate" refers to a physical association of a compound of the invention with one or more solvent molecules, whether organic or inorganic. This physical association typically includes hydrogen bonding. In some cases, the solvate is capable of separation, for example, when one or more solvate molecules are incorporated into the crystal lattice of a crystalline solid. "Solvate" encompasses both solution phase and separable solvates. Exemplary solvates include hydrates, ethanolates, and methanolates.

[0069] As used herein, the term "therapeutically effective amount" refers to an amount of a therapeutic agent sufficient to cause an improvement in one or more symptoms of a disorder, or to prevent the progression of a disorder, or to cause regression of a disorder. For example, with respect to the treatment of cancer, in one embodiment, a therapeutically effective amount will refer to an amount of a therapeutic agent that reduces tumor growth rate, reduces tumor mass, reduces the number of metastases, increases the time to tumor progression, or increases survival time by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 100%.

[0070] As used herein, the terms "sensitize" and "sensitizing" refer to making an animal or a cell in an animal body more sensitive or responsive to the biological effects of a second agent (e.g., aspects of promoting or blocking cell functions, including but not limited to cell division, cell growth, proliferation, invasion, angiogenesis, necrosis or apoptosis) by administering a first agent (e.g., a benzoic acid compound of the present invention). The sensitization of a target cell by a first agent can be measured as the difference in the expected biological effects observed when a second agent is administered with and without the first agent (e.g., aspects of promoting or blocking cell functions, including but not limited to cell growth, proliferation, invasion, angiogenesis or apoptosis). The response of the sensitized cells can be increased by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 100%, at least about 150%, at least about 200%, at least about 250%, at least 300%, at least about 350%, at least about 400%, at least about 450%, or at least about 500% over the response in the absence of the first agent.

[0071] As used herein, the term "dysregulation of apoptosis" refers to any aberration in the ability (e.g., propensity) of a cell to undergo cell death via apoptosis. Dysregulation of apoptosis is associated with or induced by a variety of conditions, non-limiting examples of which include autoimmune disorders (e.g., systemic lupus erythematosus, rheumatoid arthritis, graft-versus-host disease, myasthenia gravis, or Sjögren's syndrome ( syndrome), chronic inflammatory disorders (e.g., psoriasis, asthma, or Crohn's disease), hyperproliferative disorders (e.g., tumors, B-cell lymphomas, or T-cell lymphomas), viral infections (e.g., herpes, papilloma, or HIV), and other disorders such as osteoarthritis and atherosclerosis.

[0072] As used herein, the term "hyperproliferative disease" refers to any condition in which a localized proliferating cell population in an animal is not subject to the usual restrictions of normal growth. Examples of hyperproliferative disorders include tumors, neoplasms, lymphomas, etc. If a neoplasm does not undergo invasion or metastasis, it is called benign and if it undergoes one of the two, it is called malignant. "Metastatic" cells mean that the cells can invade and destroy adjacent body structures. Hyperplasia is a form of cell proliferation involving an increase in the number of cells in a tissue or organ without significant changes in structure or function. Metaplasia is a form of controlled cell growth in which one type of fully differentiated cell replaces another type of differentiated cell.

[0073] The pathological growth of activated lymphoid cells usually leads to autoimmune diseases or chronic inflammatory diseases. As used herein, the term "autoimmune disease" refers to any disease in which an organism produces antibodies or immune cells that recognize its own molecules, cells or tissues. Non-limiting examples of autoimmune diseases include autoimmune hemolytic anemia, autoimmune hepatitis, Berger's disease or IgA nephropathy, celiac sprue, chronic fatigue syndrome, Crohn's disease, dermatomyositis, fibromyalgia, graft-versus-host disease, Graves' disease, Hashimoto's thyroiditis, idiopathic thrombocytopenic purpura, lichen planus, multiple sclerosis, myasthenia gravis, psoriasis, rheumatic fever, rheumatoid arthritis, scleroderma, Sjögren's syndrome, systemic lupus erythematosus, type 1 diabetes, ulcerative colitis, vitiligo, etc.

[0074] As used herein, the term "neoplastic disease" refers to any abnormal growth of benign (non-cancerous) or malignant (cancerous) cells.

[0075] As used herein, the term "normal cell" refers to a cell that is not undergoing abnormal growth or division. Normal cells are noncancerous and are not part of any hyperproliferative disease or disorder.

[0076] As used herein, the term "anti-neoplastic agent" refers to any compound that slows the proliferation, growth, or spread of a targeted (eg, malignant) neoplasm.

[0077] As used herein, the terms "prevent," "preventing," and "prevention" refer to a reduction in the presence of pathological cells (e.g., hyperproliferative or neoplastic cells) in an animal. Prevention can be complete, e.g., the complete absence of pathological cells in a subject. Prevention can also be partial, such that the presence of pathological cells in a subject is less than would have occurred in a subject without the present invention.

[0078] The term "pharmaceutically acceptable carrier" or "pharmaceutically acceptable vehicle" includes any standard pharmaceutical carrier, solvent, surfactant or vehicle. Suitable pharmaceutically acceptable carriers include aqueous carriers and non-aqueous carriers. Standard pharmaceutical carriers and their formulations are described in Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, PA, 19th ed. 1995.

[0079] Experiments conducted during the development process synthesized a variety of new small molecules with bicyclic and tetracyclic quinone scaffolds that have improved metabolic stability, solubility, and pharmacokinetic properties. Indeed, as described in Example III, various compounds of the present invention were shown to inhibit the growth of various cancer cell lines (see Tables 2, 3, 4, and 5; Figure 1 , 2 , 3, 4 and 5).

[0080] Thus, the present invention provides a new class of small molecules having bicyclic and tetracyclic quinone structures with improved metabolic stability, solubility and pharmacokinetic properties. The present invention further provides the use of such small molecules as therapeutic agents for the treatment of cancer.

[0081] In a specific embodiment, compounds having bicyclic and tetracyclic quinone structures (or analogs) contained within Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV, XVI and XVII are provided:

[0082]

[0083]

[0084] This includes pharmaceutically acceptable salts, solvates and / or prodrugs thereof.

[0085] In some embodiments, R 1 Selected from the group consisting of hydrogen, halogen, alkyl, heteroalkyl, optionally substituted C6-C 14 aryl and optionally substituted 5- to 14-membered heteroaryl.

[0086] In some embodiments, R 1 Selected from the group consisting of: H,

[0087] In some embodiments, R 2 Selected from the group consisting of alkyl, optionally substituted C4-C8 cycloalkyl, optionally substituted C4-C8 heteroalkyl, optionally substituted C4-C8 heterocycloalkyl, optionally substituted C5-C6 aryl and optionally substituted C4-C6 heteroaryl.

[0088] In some embodiments, R 2 Optionally selected from the group consisting of:

[0089]

[0090] In some embodiments, R 3a , R 3b, R 3c or R 3d Selected from the group consisting of hydrogen, halogen, methyl, methoxy, trifluoromethyl, hydroxyl or cyano.

[0091] In some embodiments, R 3a , R 3b , R 3c or R 3d is optionally mono- or poly-substituted and selected from the group consisting of:

[0092]

[0093] In some embodiments, R 4 Selected from the group consisting of alkyl or heteroalkyl.

[0094] In some embodiments, R 4 Optionally selected from the group consisting of:

[0095]

[0096] In some embodiments, A includes cycloalkyl, heterocycloalkyl, aryl, or heteroaryl.

[0097] In some embodiments, X is selected from the group consisting of hydrogen, halogen, amino, heterocycloalkyl, or hydroxy.

[0098] In some embodiments, X is optionally selected from the group consisting of:

[0099]

[0100] In some embodiments, n is an integer selected from 0 to 5.

[0101] In some embodiments, m is an integer selected from 0 to 9.

[0102] In some embodiments, Y is independently selected from O, S, and N atoms.

[0103] In some embodiments, W is independently selected from C atoms, N atoms.

[0104] In some embodiments, for Formulas I-XVII, the compounds shown in Table 1 are contemplated.

[0105] In some embodiments, for Formula I, the compounds shown in Table 1 are contemplated.

[0106] An important aspect of the present invention is that the compounds of the present invention induce cell cycle arrest and / or apoptosis alone or in response to additional apoptosis inducing signals and also strengthen the induction of cell cycle arrest and / or apoptosis. Therefore, it is expected that these compounds sensitize cells to the induction of cell cycle arrest and / or apoptosis, including cells resistant to such induction stimuli. The compounds of the present invention (e.g., indole (or analog) compounds) can be used to induce apoptosis in any disease that can be treated, improved or prevented by inducing apoptosis. In one embodiment, a modulator (e.g., an inhibitor) can be used to induce apoptosis in cells by inducing reactive oxygen species in one or more cells of interest.

[0107] In some embodiments, the compositions and methods of the present invention are used to treat sick cells, tissues, organs or pathological conditions and / or disease states in animals (e.g., mammalian patients, including but not limited to humans and veterinary animals). In this regard, the methods and compositions of the present invention are used to treat or prevent various diseases and pathological changes. A non-limiting exemplary list of such diseases and conditions includes, but is not limited to, pancreatic cancer, breast cancer, prostate cancer, lymphoma, skin cancer, colon cancer, melanoma, malignant melanoma, ovarian cancer, brain cancer, primary brain carcinoma, head and neck cancer, glioma, glioblastoma, liver cancer, bladder cancer, non-small cell lung cancer, head or neck carcinoma, breast carcinoma, ovarian carcinoma, lung carcinoma, small-cell lung carcinoma, Wilms' tumor, cervical carcinoma, testicular carcinoma, bladder carcinoma, pancreatic carcinoma, stomach cancer, carcinoma, colon carcinoma, prostatic carcinoma, genitourinary carcinoma, thyroid carcinoma, esophageal carcinoma, myeloma, multiple myeloma, adrenal carcinoma, renal cell carcinoma, endometrial carcinoma, adrenal cortex carcinoma, malignant pancreatic insulinoma, malignant carcinoid carcinomacarcinoma), choriocarcinoma, mycosis fungoides, malignant hypercalcemia, cervical hyperplasia, leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, chronic granulocytic leukemia, acute myeloid leukemia, hairy cell leukemia, neuroblastoma, rhabdomyosarcoma, Kaposi's sarcoma, polycythemia vera, essential thrombocytosis, Hodgkin's disease, non-Hodgkin's lymphoma, soft tissue sarcoma, osteogenic sarcoma, essential macroglobulinemia and retinoblastoma, T and B cell-mediated autoimmune diseases; inflammatory diseases; infections; hyperproliferative diseases; AIDS; degenerative diseases, vascular diseases, etc. In some embodiments, the cancer cells treated are metastatic. In other embodiments, the cancer cells treated are resistant to anti-cancer agents.

[0108] Some embodiments of the present invention provide methods for administering an effective amount of a compound of the present invention and at least one additional therapeutic agent (including but not limited to chemotherapeutic anti-neoplastic agents, apoptosis modulators, antimicrobial agents, antiviral agents, antifungal agents and anti-inflammatory agents) and / or therapeutic techniques (e.g., surgical intervention and / or radiation therapy). In specific embodiments, one or more additional therapeutic agents are anti-cancer agents.

[0109] Many suitable anti-cancer agents are contemplated for use in the methods of the present invention. In fact, the present invention contemplates, but is not limited to, the administration of a variety of anti-cancer agents, such as: agents that induce apoptosis; polynucleotides (e.g., antisense, ribozymes, siRNA); polypeptides (e.g., enzymes and antibodies); biomimetic substances; alkaloids; alkylating agents; antitumor antibiotics; antimetabolites; hormones; platinum compounds; monoclonal or polyclonal antibodies (e.g., antibodies conjugated to anticancer drugs, toxins, defensins), toxins; radionuclides; biological response modifiers (e.g., interferons (e.g., IFN-α) and interleukins (e.g., IL-2)); adoptive immunotherapy agents; hematopoietic growth factors; agents that induce tumor cell differentiation (e.g., all-trans retinoic acid); gene therapy agents (e.g., antisense therapeutic agents and nucleotides); tumor vaccines; angiogenesis inhibitors; proteasome inhibitors: NF-КB modulators; anti-CDK compounds; HDAC inhibitors; etc. Many other examples of chemotherapeutic compounds and anti-cancer therapies suitable for co-administration with the disclosed compounds are known to those skilled in the art.

[0110] In certain embodiments, the anticancer agent includes an agent that induces or stimulates apoptosis. Agents that induce apoptosis include, but are not limited to, radiation (e.g., X-rays, gamma rays, UV); tumor necrosis factor (TNF)-related factors (e.g., TNF family receptor proteins, TNF family ligands, TRAIL, antibodies to TRAIL-R1 or TRAIL-R2); kinase inhibitors (e.g., epidermal growth factor receptor (EGFR) kinase inhibitors, vascular growth factor receptor (VGFR) kinase inhibitors, fibroblast growth factor receptor (FGFR) kinase inhibitors, platelet-derived growth factor receptor (PDGFR) kinase inhibitors); agents, and Bcr-Abl kinase inhibitors (such as GLEEVEC); antisense molecules; antibodies (e.g., HERCEPTIN, RITUXAN, ZEVALIN, and AVASTIN); antiestrogens (e.g., raloxifene and tamoxifen); antiandrogens (e.g., flutamide, bicalutamide, finasteride, aminoglutethamide, ketoconazole, and corticosteroids); cyclooxygenase 2 (COX-2) inhibitors (e.g., celecoxib, meloxicam, NS-398, and nonsteroidal antidepressants). nonsteroidal anti-inflammatory drugs (NSAIDs); anti-inflammatory drugs (for example, butazolidin, DECADRON, DELTASONE, dexamethasone, dexamethasone intensol, DEXONE, HEXADROL, hydroxychloroquine, METICORTEN, ORADEXON, ORASONE, hydroxybutazone, PEDIAPRED, phenylbutazone, PLAQUENIL, prednisolone, prednisone, PRELONE, and TAND EARIL); and cancer chemotherapy drugs (e.g., irinotecan (CAMPTOSAR), CPT-11, fludarabine (FLUDARA), dacarbazine (DTIC), dexamethasone, mitoxantrone, MYLOTARG, VP-16, cisplatin, carboplatin, oxaliplatin, 5-FU, doxorubicin, gemcitabine, bortezomib, gefitinib, bevacizumab, TAXOTERE or TAXOL); cell signaling molecules; ceramides and cytokines; staurosporine, etc.

[0111] In yet other embodiments, the compositions and methods of the invention provide a compound of the invention and at least one anti-hyperproliferative or anti-neoplastic agent selected from alkylating agents, antimetabolites, and natural products (e.g., herbs and other plant and / or animal derived compounds).

[0112] Alkylating agents suitable for use in the compositions and methods of the invention include, but are not limited to: 1) nitrogen mustards (e.g., mechlorethamine, cyclophosphamide, ifosfamide, melphalan (L-sabolesine); and chlorambucil); 2) ethyleneimines and methylmelamines (e.g., hexamethylmelamine and thiotepa); 3) alkyl sulfonates (e.g., busulfan); 4) nitrosoureas (e.g., carmustine (BCNU); lomustine (CCNU); semustine (methyl-CCNU); and streptozotocin (streptozotocin); and 5) triazenes (e.g., dacarbazine (DTIC; dimethyltriazenylimidazolecarboxamide).

[0113] In some embodiments, antimetabolites suitable for use in the compositions and methods of the present invention include, but are not limited to: 1) folic acid analogs (e.g., methotrexate (methotrexate)); 2) pyrimidine analogs (e.g., fluorouracil (5-fluorouracil; 5-FU), floxuridine (fluorodeoxyuridine; FudR) and cytarabine (cytosine arabinoside)); and 3) purine analogs (e.g., mercaptopurine (6-mercaptopurine; 6-MP), thioguanine (6-thioguanine; TG) and pentostatin (2'-deoxycoformycin)).

[0114] In yet other embodiments, chemotherapeutic agents suitable for use in the compositions and methods of the present invention include, but are not limited to: 1) vinca alkaloids (e.g., vinblastine (VLB), vincristine); 2) epipodophyllotoxins (e.g., etoposide and teniposide); 3) antibiotics (e.g., dactinomycin (actinomycin D), daunorubicin (daunorubicin; erythromycin), doxorubicin, bleomycin, plicamycin (mithramycin) and mitomycin (mitomycin C)); 4) enzymes (e.g., L-asparaginase); 5) biological response modifiers (e.g., interferon-α); 6) platinum coordination complexes (e.g., cisplatin (cis-DDP) and carboplatin); 7) anthracenediones (e.g., mitoxantrone); 8) substituted ureas (e.g., hydroxyurea); 9) methylhydrazine derivatives (e.g., procarbazine (N-methylhydrazine; MIH)); 10) adrenocortical suppressants (e.g., mitotane (o,p'-DDD) and aminoglutethimide); 11) corticosteroids (e.g., prednisone); 12) progestins (e.g., hydroxyprogesterone caproate, medroxyprogesterone acetate, and megestrol acetate); 13) estrogens (e.g., diethylstilbestrol and ethinyl estradiol); 14) antiestrogens (e.g., tamoxifen); 15) androgens (e.g., testosterone propionate and fluoxymesterone); 16) antiandrogens (e.g., flutamide); and 17) gonadotropin-releasing hormone analogs (e.g., leuprolide).

[0115] Any oncolytic agent routinely used in the context of cancer treatment is used in the compositions and methods of the present invention. For example, the U.S. Food and Drug Administration (US Food and Drug Administration) maintains a formulary of oncolytic agents approved for use in the United States. International counterparts to the U.S. Food and Drug Administration (USFDA) maintain similar formularies. Table 6 provides a list of exemplary anti-neoplastic agents approved for use in the United States. Those skilled in the art will appreciate that the "product label" required on all U.S.-approved chemotherapeutic drugs describes the approved indications, dosage information, toxicity data, etc. for the exemplary agents.

[0116] Table 6

[0117]

[0118]

[0119]

[0120]

[0121]

[0122]

[0123]

[0124]

[0125]

[0126]

[0127]

[0128]

[0129] Anticancer agents further include compounds that have been identified to have anticancer activity. Examples include, but are not limited to, 3-AP, 12-O-tetradecanoylphorbol-13-acetate, 17AAG, 852A, ABI-007, ABR-217620, ABT-751, ADI-PEG20, AE-941, AG-013736, AGRO100, Aladenosine, AMG 706, Antibody G250, Antineoplastons, AP23573, Apaquinone, APC8015, Atimod, ATN-161, Atrasenten, Azacitidine, BB-10901, BCX-1777, Bevacizumab, BG00001, Bicalutamide, BMS247550, Bortezomib, Bryostatin-1, Buserelin, Calcitriol, CCI-779, CDB-2914, Cefixime, Cetuximab, CG0070, Cilengitide, Clofarabine, Combretastatin A4 Phosphate, CP-675,206, CP-724,714, CpG 7909, curcumin, decitabine, DENSPM, doxorcalciferol, E7070, E7389, ecteinascidin 743, efacile, eflornithine, EKB-569, enzastaurin, erlotinib, exisulin, fenretinide, flavopiridol, fludarabine, flutamide, fotemustine, FR901228, G17DT, galiximab, gefitinib, genistein, glufosfamide, GTI-2040, histrelin, HKI-272, homoharringtonine, HSPPC-96, hu14.18-interleukin-2 fusion protein, HuMax-CD4, iloprost, imiquimod, infliximab, interleukin-12, IPI-504, irovin, ixabepilone, lapatinib, lenalidomide, letutinib, leuprolide, LMB-9 immunotoxin, lonafarnib, luniliximab, mafosfamide, MB07133, MDX-010, MLN2704, monoclonal antibody 3F8, monoclonal antibody J591, motexa fen, MS-275, MVA-MUC1-IL2, nilutamide, nitrocamptothecin, nolatriptide dihydrate, tamoxifen (nolvadex), NS-9, O6-benzylguanine, olimusen sodium, ONYX-015, ogavumab, OSI-774, panitumumab, paraplatin, PD-0325901, pemetrexed, PHY906, pioglitazone, pirfenidone, pixanol, PS-341, PSC 833, PXD101, pyrazoloacridine, R115777, RAD001, ranpirnase, rebeccamycin analogs, rhuAngiostatin protein, rhuMab 2C4, rosiglitazone, rubitecan, S-1, S-8184, satraplatin, SB-, 15992, SGN-0010, SGN-40, sorafenib, SR31747A, ST1571, SU011248, suberoylanilide hydroxamic acid, suramin, taporostat, talampanel, talizoloft, temsirolimus, TGFa-PE38 immunotoxin, thalidomide, thymalfasin, tipifarnib, tirapazamine, TLK286, trabectedin, trimetrexate glucuronide, TroVax, UCN-1, valproic acid, vinflunine, VNP40101M, voroxizumab, vorinrestat, VX-680, ZD1839, ZD6474, zileuton, and zosuquidar trihydrochloride. .

[0130] For a more detailed description of anti-cancer agents and other therapeutic agents, those skilled in the art are referred to any number of instruction manuals, including but not limited to the Physician's Desk Reference and Goodman and Gilman's "Pharmaceutical Basis of Therapeutics" tenth edition, Eds. Hardman et al., 2002.

[0131] The present invention provides methods for administering the compounds of the present invention with radiotherapy. The present invention is not limited by the type, amount, or delivery and administration system used to deliver a therapeutic dose of radiation to an animal. For example, an animal can receive photon radiotherapy, particle beam radiotherapy, other types of radiotherapy, and combinations thereof. In some embodiments, radiation is delivered to an animal using a linear accelerator. In yet other embodiments, radiation is delivered using a gamma knife.

[0132] The radiation source can be outside or inside the animal. External radiation therapy is the most common, and involves using, for example, a linear accelerator to guide a beam of high-energy radiation through the skin to the tumor site. Although the radiation beam is positioned at the tumor site, it is almost impossible to avoid exposing normal, healthy tissue. However, animals usually tolerate external radiation well. Internal radiation therapy involves implanting a radiation emission source, such as beads, wires, pills, capsules, particles, etc., at or near the tumor site in the body, including the use of a delivery system that specifically targets cancer cells (for example, using particles attached to a cancer cell binding ligand). Such implants can be removed after treatment, or left in the body without activity. The types of internal radiation therapy include, but are not limited to, brachytherapy, interstitial irradiation, intracavitary irradiation, radioimmunotherapy, etc.

[0133] Animals may optionally receive a radiosensitizer (e.g., metronidazole, misonidazole, intra-arterial BudR, intravenous iodideoxyuridine (IudR), nitroimidazoles, 5-substituted-4-nitroimidazoles, 2H-isoindolediones, [[(2-bromoethyl)-amino]methyl]-nitro-1H-imidazole-1-ethanol, nitroaniline derivatives, DNA-affinic hypoxia selective cytotoxins, Cytotoxins), halogenated DNA ligands, 1,2,4-benzotriazine oxides, 2-nitroimidazole derivatives, fluorine-containing nitroazole derivatives, benzamides, nicotinamides, acridine-intercalators, 5-thiotetrazole derivatives, 3-nitro-1,2,4-triazoles, 4,5-dinitroimidazole derivatives, hydroxylated texaphrins, cisplatin, mitomycin, tiripazamine, nitrosoureas, mercaptopurines, methotrexate, fluorouracil, bleomycin, vincristine, carboplatin, epirubicin, doxorubicin, cyclophosphamide, vindesine, etoposide, paclitaxel, heat (hyperthermia), etc.), radioprotectants (e.g., cysteamine, aminoalkylthiophosphate dihydrogen salts, amifostine (WR 2721), IL-1, IL-6, etc.). Radiosensitizers enhance the killing of tumor cells. Radioprotectants protect healthy tissue from the harmful effects of radiation.

[0134] Any type of radiation can be administered to animals, as long as the animal tolerates the radiation dose without unacceptable negative side effects. Suitable types of radiotherapy include, for example, ionizing (electromagnetic) radiotherapy (e.g., X-rays or gamma rays) or particle beam radiotherapy (e.g., high line energy radiation). Ionizing radiation is defined as radiation including particles or photons, which have enough energy to produce ionization, i.e., the gain or loss of electrons (as described in, for example, US5,770,581, which is incorporated herein by reference in its entirety). The effect of radiation can be controlled at least in part by a clinician. In one embodiment, for maximum target cell exposure and reduced toxicity, the radiation dose is divided into several parts.

[0135] In one embodiment, the total dose of radiation administered to an animal is from about .01 gray (Gy) to about 100 Gy. In another embodiment, about 10 Gy to about 65 Gy (e.g., about 15 Gy, 20 Gy, 25 Gy, 30 Gy, 35 Gy, 40 Gy, 45 Gy, 50 Gy, 55 Gy, or 60 Gy) is administered during treatment. Although in some embodiments a full dose of radiation can be administered over the course of a day, the total dose is ideally divided into several portions and administered over several days. Ideally, radiation therapy is administered over the course of at least about 3 days, for example, at least 5, 7, 10, 14, 17, 21, 25, 28, 32, 35, 38, 42, 46, 52, or 56 days (about 1-8 weeks). Therefore, the daily dose of radiation will include about 1-5Gy (e.g., about 1Gy, 1.5Gy, 1.8Gy, 2Gy, 2.5Gy, 2.8Gy, 3Gy, 3.2Gy, 3.5Gy, 3.8Gy, 4Gy, 4.2Gy or 4.5Gy) or 1-2Gy (e.g., 1.5-2Gy). The daily dose of radiation should be sufficient to induce the destruction of target cells. If extended for a period of time, in one embodiment, radiation is not administered every day, thereby allowing the animal to rest and achieve the effect of treatment. For example, for weekly treatment, radiation is ideally administered for 5 consecutive days, and radiation is not administered on 2 days, thereby allowing 2 days of rest per week. However, radiation can be administered 1 day / week, 2 days / week, 3 days / week, 4 days / week, 5 days / week, 6 days / week or all 7 days / week, depending on the responsiveness of the animal and any potential side effects. Radiotherapy can start at any time during the treatment period. In one embodiment, radiation is initiated in week 1 or week 2 and is administered for the remainder of the treatment period. For example, radiation is administered in weeks 1-6 or weeks 2-6 of a treatment period that includes 6 weeks for treating, for example, solid tumors. Alternatively, radiation is administered in weeks 1-5 or weeks 2-5 of a treatment period that includes 5 weeks. However, these exemplary radiation therapy dosing schedules are not intended to limit the present invention.

[0136] Antimicrobial therapeutic agents can also be used as therapeutic agents in the present invention. Any agent that can kill, inhibit or otherwise reduce the function of a microbial organism can be used, as well as any agent that is expected to have such activity. Antimicrobial agents include, but are not limited to, natural and synthetic antibiotics, antibodies, inhibitory proteins (e.g., defensins), antisense nucleic acids, membrane disruptors, etc., used alone or in combination. In fact, any type of antibiotic can be used, including but not limited to antibacterial agents, antiviral agents, antifungal agents, etc.

[0137] In some embodiments of the invention, the compounds of the invention and one or more therapeutic or anti-cancer agents are administered to animals under one or more of the following conditions: at different cycles, different durations, different concentrations, different routes of administration, etc. In some embodiments, the compound is administered before the therapeutic or anti-cancer agent, for example, 0.5, 1, 2, 3, 4, 5, 10, 12 or 18 hours, 1, 2, 3, 4, 5 or 6 days, or 1, 2, 3 or 4 weeks before the administration of the therapeutic or anti-cancer agent. In some embodiments, the compound is administered after the therapeutic or anti-cancer agent, for example, 0.5, 1, 2, 3, 4, 5, 10, 12 or 18 hours, 1, 2, 3, 4, 5 or 6 days, or 1, 2, 3 or 4 weeks after the administration of the anti-cancer agent. In some embodiments, the compound and the therapeutic or anti-cancer agent are administered simultaneously but in different schedules, for example, the compound is administered daily, while the therapeutic or anti-cancer agent is administered once a week, once every two weeks, once every three weeks, or once every four weeks. In other embodiments, the compound is administered once a week, while the therapeutic or anti-cancer agent is administered daily, weekly, once every two weeks, once every three weeks, or once every four weeks.

[0138] Compositions within the scope of the present invention include all compositions containing the compounds of the present invention in an amount that effectively achieves its intended purpose. Although individual needs vary, it is within the technical scope of the art to determine the optimal range of the effective amount of each component. Typically, these compounds or their pharmaceutically acceptable salts of equivalent amounts can be orally administered to mammals, such as humans, at a dose of 0.0025 to 50 mg / kg of the body weight of a mammal treated for an illness that responds to apoptosis induction every day. In one embodiment, oral administration of about 0.01 to about 25 mg / kg is to treat, improve or prevent such an illness. For intramuscular injection, the dosage is generally about half of the oral dosage. For example, a suitable intramuscular dose will be about 0.0025 to about 25 mg / kg, or about 0.01 to about 5 mg / kg.

[0139] The unit oral dose may include about 0.01 to about 1000 mg, for example, about 0.1 to about 100 mg of the compound. The unit dose may be administered once or more per day as one or more tablets or capsules, each tablet or capsule containing about 0.1 to about 10 mg, conveniently about 0.25 to 50 mg of the compound or its solvate.

[0140] In topical formulations, the compound may be present in a concentration of about 0.01 to 100 mg / gram of carrier. In one embodiment, the compound is present in a concentration of about 0.07-1.0 mg / ml, such as about 0.1-0.5 mg / ml, and in one embodiment, is present in a concentration of about 0.4 mg / ml.

[0141] In addition to administering the compounds as raw chemicals, the compounds of the invention may also be administered as part of a pharmaceutical preparation containing suitable pharmaceutically acceptable carriers including excipients and adjuvants that facilitate processing of the compounds into preparations that can be used pharmaceutically. Preparations, particularly those that can be administered orally or topically and can be used in one type of administration, such as tablets, dragees, sustained-release lozenges and capsules, mouthrinses and mouthwashes, gels, liquid suspensions, hair rinses, hair gels, shampoos, and also formulations that can be administered rectally, such as suppositories, and suitable solutions for intravenous infusion, injection, topical or oral administration, contain from about 0.01% to 99%, in one embodiment from about 0.25% to 75% of one or more active compounds and excipients.

[0142] The pharmaceutical compositions of the present invention can be administered to any patient who can experience the beneficial effects of the compounds of the present invention. Among such patients, the most important are mammals, such as humans, although the present invention is not intended to be so limited. Other patients include veterinary animals (cattle, sheep, pigs, horses, dogs, cats, etc.).

[0143] Compounds and pharmaceutical compositions thereof can be administered in any manner that achieves their intended purpose. For example, administration can be by parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, buccal, intrathecal, intracranial, intranasal or topical routes. Alternatively or simultaneously, administration can be by oral route. The dosage administered will depend on the age, health and weight of the recipient, the type of concurrent treatment (if any), the frequency of treatment and the nature of the desired effect.

[0144] The pharmaceutical preparations according to the invention are manufactured in a manner known per se, for example, by means of conventional mixing, granulating, dragee-making, dissolving or lyophilizing processes. Thus, pharmaceutical preparations for oral use can be obtained by combining the active compounds with solid excipients, optionally grinding the resulting mixture after adding suitable auxiliaries, if desired or necessary, and processing the mixture of granules to obtain tablets or dragee cores.

[0145] Suitable excipients are fillers in particular, such as, sugar, for example lactose or sucrose, mannitol or sorbitol, cellulose preparations and / or calcium phosphate, for example tricalcium phosphate or calcium hydrogen phosphate, and binding agents, such as starch slurry, use for example, corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth, methylcellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose and / or polyvinyl pyrrolidone. If desired, disintegrants can be added, such as above-mentioned starch, and also carboxymethyl starch, cross-linked polyvinyl pyrrolidone, agar or alginic acid or its salt, such as sodium alginate. Auxiliary agents, especially flow regulators and lubricants, for example silicon dioxide, talcum, stearic acid or its salt, such as magnesium stearate or calcium stearate, and / or Polyethylene Glycol. Dragee cores are provided with suitable coatings, and if desired, they resist gastric juice. For this purpose, concentrated sugar solution can be used, which can optionally comprise gum arabic, talcum, polyvinyl pyrrolidone, polyethylene glycol and / or titanium dioxide, lacquer solution and suitable organic solvent or solvent mixture. In order to produce the coating against gastric juice, a solution of suitable cellulose preparations such as acetylcellulose phthalate or hydroxypropyl methylcellulose phthalate is used. Dye or pigment can be added to tablet or dragee coating, for example, for identification or in order to characterize the combination of active compound dosage.

[0146] Other pharmaceutical preparations that can be used orally include push-fit capsules made of gelatin, and soft sealed capsules made of gelatin and plasticizers such as glycerol or sorbitol. The push-fit capsules can include active compounds in the form of particles, which can be mixed with fillers (such as lactose), binders (such as starch) and / or lubricants (such as talc or magnesium stearate) and optionally stabilizers. In soft capsules, in one embodiment, the active compound is dissolved or suspended in a suitable liquid, such as a fatty oil or liquid paraffin. In addition, stabilizers can be added.

[0147] Possible pharmaceutical preparations that can be used rectally include, for example, suppositories, which are composed of a combination of one or more active compounds and a suppository base. Suitable suppository bases are, for example, natural or synthetic triglycerides or paraffin hydrocarbons. In addition, gelatin rectal capsules composed of a combination of active compounds and a base may also be used. Possible base materials include, for example, liquid triglycerides, polyethylene glycols or paraffin hydrocarbons.

[0148] Suitable formulations for parenteral administration include aqueous solutions of the active compound in water-soluble form, such as water-soluble salts and alkaline solutions. In addition, a suspension of the active compound as a suitable oily injection suspension can be administered. Suitable lipophilic solvents or vehicles include fatty oils, such as sesame oil or synthetic fatty acid esters, such as ethyl oleate or triglycerides or polyethylene glycol-400. Aqueous injection suspensions can include materials that increase the viscosity of the suspension, including, for example, sodium carboxymethylcellulose, sorbitol and / or dextran. Optionally, the suspension can also include a stabilizer.

[0149] In one embodiment, the topical composition of the present invention is formulated into oils, creams, lotions, ointments, etc. by selecting appropriate carriers. Suitable carriers include vegetable oils or mineral oils, white petrolatum (white soft paraffin), branched chain fats or oils, animal fats and high molecular weight alcohols (greater than C). 12 ). The carrier may be one in which the active ingredient is soluble. If desired, emulsifiers, stabilizers, wetting agents and antioxidants as well as agents that impart color or fragrance may also be included. In addition, transdermal permeation enhancers may be used in these topical preparations. Examples of such enhancers may be found in U.S. Pat. Nos. 3,989,816 and 4,444,762; each of which is incorporated herein by reference in its entirety.

[0150] Ointments can be prepared by mixing a solution of the active ingredient in a vegetable oil (such as almond oil) with warm soft paraffin and allowing the mixture to cool. A typical example of such an ointment is an ointment comprising about 30% almond oil and about 70% white soft paraffin by weight. Lotions can be conveniently prepared by dissolving the active ingredient in a suitable high molecular weight alcohol such as propylene glycol or polyethylene glycol.

[0151] Those skilled in the art will readily appreciate that the foregoing only represents a detailed description of certain preferred embodiments of the present invention. Various modifications and variations of the above compositions and methods can be readily accomplished using the expertise available in the art, and they are all within the scope of the present invention.

[0152] Example

[0153] The following examples are illustrative of the compounds, compositions and methods of the present invention, but are not limiting. Other suitable modifications and adaptations of the various conditions and parameters normally encountered in clinical therapy and obvious to those skilled in the art are within the spirit and scope of the present invention.

[0154] Example I.

[0155] This example describes various synthetic methods used to obtain compounds of the present invention.

[0156] Methods for preparing bicyclic quinone derivatives of Formula I are shown in Scheme 1 and Scheme 2.

[0157] Solution 1 a Synthesis of compound Ia.

[0158]

[0159] a Reagents and conditions: (a) NaClO3, concentrated HCl, room temperature -50°C, 2h; (b) aromatic amine, CeCl3·7H2O, EtOH, 60-90°C, 2-6h;

[0160] Where R 1 , R 2 , R 3a , R 3b , R 3c and R 3d Same as defined above.

[0161] The detailed synthetic method in Scheme 1 is described below.

[0162] The compound of formula Ia is prepared as follows (Scheme 1): 8-Hydroxyquinoline derivative 1-1 is chlorinated with sodium chlorate in the presence of concentrated hydrochloric acid under simple magnetic stirring to obtain 6,7-dichloro-5,8-quinolinedione 1-2. The compound of formula I is prepared by condensation reaction of 6,7-dichloro-5,8-quinolinedione 1-2 with aromatic amine. The solvent used in the reaction is C1-C3 lower alcohol, such as methanol, ethanol and isopropanol. In the presence of Lewis acid, such as cerium (III) chloride heptahydrate (CeCl7·7H2O), nickel (II) chloride hexahydrate (NiCl2·6H2O) and iron (III) chloride hexahydrate FeCl3·6H2O, an additive is used in this regioselective condensation of the appropriate aminobenzene to the 6 position of 6,7-dichloroquinoline-5,8-dione 1-2. The reaction is carried out at reflux temperature for 2-6 hours.

[0163] Solution 2 a Synthesis of compound Ib.

[0164]

[0165] a Reagents and conditions: (c) KOH, MeOH, H2O, 80-100°C, 2-5 hours.

[0166] Where R 1 and R 2 Same as defined above.

[0167] The detailed synthetic method in Scheme 2 is described below.

[0168] The compound of formula Ib is prepared by reacting compound I-a' dissolved in methanol and water with potassium hydroxide at 80°C for 2 hours (Scheme 2).

[0169] The method for preparing the bicyclic quinone derivatives of Formula II is shown in Scheme 3. The detailed synthetic method is described below.

[0170] Solution 3 a Synthesis of compound II.

[0171]

[0172] a Reagents and conditions: (a) Meldrum's acid, methyl orthoformate, reflux, 7h; (b) HNO3 / SiO2, CH2Cl2, room temperature; (c) PhOPh, 250℃, 15min; (d) POBr3, DMF, 0℃-room temperature; (e) substituted boronic acid, Pd(PPh3)4, K2CO3, dioxane, H2O, reflux, 6h; (f) Fe, AcOH, MeOH, H2O, reflux, 30min; (g) acyl chloride, TEA, THF, 0℃-room temperature, overnight; (h) (NH4)2Ce(NO3)6, CH3CN, H2O, 0℃-room temperature, 2h; (i) amine, dry chloroform, room temperature, 0.5-8h; (j) amine, CHCl3, reflux, 4h.

[0173] Where R 1 , R 4 and X are the same as defined above.

[0174] The detailed synthetic method in Scheme 3 is described below.

[0175] The preparation of compounds of formula II-a and formula II-b is depicted as follows: The synthesis of Ic is depicted in Scheme 3. Nitroquinolone 1-7 is synthesized by a well-established method (Heterocycl. Commun. 2000, 6, 539-544; Synth. Commun. 1985, 15, 125-133). A solution of McLaughlin's acid is heated at reflux in methyl orthoformate for 2 h, followed by the addition of aromatic amine 1-4. The mixture is heated at reflux for another 5 h. Subsequent regioselective nitration of compound 1-5 with nitric acid supported on silica gel in dichloromethane affords 1-6. Cyclization of 1-6 to the corresponding nitroquinolone 1-7 is accomplished by boiling in diphenyl ether at 250° C. for 15 min. Treatment of quinolone 1-7 with POBr3 in DMF affords 4-bromoquinoline 1-8. Using Pd(PPh3)4 as a catalyst, intermediate 1-8 is reacted with the corresponding substituted boronic acid in dioxane by Heck coupling to produce 1-9. Subsequent reduction of the nitro group with activated iron gives the aniline product 1-10, which is then reacted with the appropriate acid chloride in the presence of trimethylamine to give 1-11. Final oxidation of 1-11 with cerium ammonium nitrate in acetonitrile and H2O gives the desired compound 1-12. Compounds of formula II-a are prepared by reacting compound 1-12 with an appropriate amine in anhydrous chloroform at 0°C to room temperature for 3-8 hours. Chloroform as a solvent can be replaced by dichloromethane, N,N-dimethylformamide or methanol. Compounds of formula II-b are prepared by reacting compound 1-12 with an appropriate amine in anhydrous chloroform and heating for 3-8 hours. Chloroform as a solvent can be replaced by dichloromethane, N,N-dimethylformamide or methanol.

[0176] A method for preparing tetracyclic quinone derivatives of formula III is shown in Scheme 4. The detailed synthetic method is described below.

[0177] Solution 4 a Synthesis of compound III.

[0178]

[0179] a Reagents and conditions: (a) NaClO3, concentrated HCl, room temperature-50°C, 2h; (b) aromatic amine, CeCl3·7H2O, EtOH, 60-80°C, 2-5h; (c) NaN3, H2O, DMF, 80–95°C, 2-8h.

[0180] Where R 1 , R 2 , R 3a , R 3b , R 3c and R 3d Same as defined above.

[0181] The detailed syntheses of the compounds in Scheme 4 are described below.

[0182] The compound of formula III is prepared as follows: 8-hydroxyquinoline derivative 2-1 is chlorinated with sodium chlorate in the presence of concentrated hydrochloric acid under simple magnetic stirring to obtain 6,7-dichloro-5,8-quinolinedione 2-2. In the presence of Lewis acid, such as cerium (III) chloride heptahydrate (CeCl7·7H2O), nickel (II) chloride hexahydrate (NiCl2·6H2O) or iron (III) chloride hexahydrate FeCl3·6H2O, 6,7-dichloro-5,8-quinolinedione I is condensed with aromatic amine. The solvent used in the reaction is C1-C3 lower alcohol, such as methanol, ethanol and isopropanol, at 60-90°C for 2-6 hours. After the reaction is completed, the solvent is removed under reduced pressure. Then sodium azide, DMF and water droplets are added. The mixture is stirred at 65-90°C for 2-6 hours to obtain the compound of formula III.

[0183] General experimental methods

[0184] General Methods. Reagents and anhydrous solvents were used without further purification and were purchased from commercial sources. The progress of the reaction was monitored by UV absorbance using thin layer chromatography (TLC) on aluminum-lined pre-coated silica plates from Silicycle (SiliaPlate, 200 μm thickness, F254). Purification using flash chromatography was performed using Silicycle silica gel (SiliaFlash F60, 40-63 μm, 230-400 mesh, PN R10030B), and small percentages of compounds were purified using a Biotage Isolera chromatography system equipped with 10 and 25 g Ultra-SNAP Cartridge columns (25 μM spherical silica). 1H NMR spectra were obtained using a Varian (300 or 400 MHz) instrument. Spectral data were reported using the following abbreviations: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, dd = doublet of doublets, and coupling constants were reported in Hz and then integrated. A Shimadzu LCMS20-20 system was used to generate HPLC traces, obtain mass spectral data, and assess purity. The system was equipped with a PDAUV detector and a Kinetex 2.6 μm, XB-C18 75 mm x 4.6 mm column, which was used at ambient temperature. The HPLC gradient method utilized: 1% to 90% MeCN in H2O with 0.01% formic acid over 20 min at a flow rate of 0.50 mL / min. The purity of the final compound was assessed at 254 nm (≥95%) using the described column and method. Reverse phase preparative purification was performed on a Shimadzu LC-20 modular HPLC system. The system utilized a PDA detector and a Kinetex 5μm XB-C18 150 mm x 21.2 mm column. The purification method used a 27 min gradient, 10% to 90% MeCN in H2O with 0.02% trifluoroacetic acid.

[0185] General Scheme A: Chloride Oxidation. Sodium chlorate (5.3 g, 50 mmol) was added to a stirred solution of 8-hydroxyquinoline derivative (10 mmol) in concentrated HCl (100 mL) at 50°C over a period of 1 h. The reaction mixture was then allowed to stir for 2 h, and then diluted to 200 mL with distilled water. The yellow precipitate formed was removed by filtration and discarded. The filtrate was extracted with CH2Cl2 (3×50 mL), and the organic phases were combined, washed with water and brine, dried (Na2SO4), and concentrated in vacuo. The solid was recrystallized in MeOH to give pure bright yellow crystalline 6,7-dichloro-5,8-quinolinediones 1-2 and 2-2.

[0186] General Scheme B: Regioselective condensation. A solution of 6,7-dichloro-5,8-quinolinedione 1-2 or 2-2 (1.0 equiv), cerium (III) chloride heptahydrate (CeCl3.7H2O, 1.1 equiv) and appropriate aromatic amine (1.0 equiv) in ethanol was stirred at 60-90° C. for 2-6 h. After completion of the reaction by TLC, it was cooled, filtered and recrystallized from ethanol to give the desired compound Ia as a dark purple powder.

[0187] General Scheme C: Hydrolysis reaction. 7-Chloro-6-((2,6-difluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)quinoline-5,8-dione 1-a' (0.66 mmol) was added to a stirred solution of KOH (0.37 g, 6.6 mmol) in MeOH:H2O (3:1) and heated to 80°C. After 2 h, the dark red solution was cooled to room temperature. 10 mL of ethyl acetate was added, washed with water (10 mL) and saturated aqueous NaCl solution (10 mL). The organic layer was dried over sodium sulfate, and then the solvent was distilled off under reduced pressure. It was purified by silica gel column chromatography to obtain the desired compound 28 (1-b).

[0188] General Scheme D: Bromination reaction.POBr3 was added dropwise to a suspension of nitroquinolone 1-7 (84 mg, 0.034 mmol) in DMF at 0°C. The suspension became clear and then turbid during this process. After 1 h, the reaction mixture was diluted with 28% ammonia solution and extracted with ethyl acetate. The combined extracts were washed with brine, dried over anhydrous Na2SO4 and removed in vacuo. The residue was purified on flash column chromatography using ethyl acetate and hexane as eluents to obtain compound 4-bromo-5,8-dimethoxy-6-nitroquinoline 1-8 as a yellow solid.

[0189] General Scheme E: Coupling reaction. Compound 1-8 (10 mg, 0.068 mmol), boronic acid (0.068 mmol) and K2CO3 (11 mg, 0.081 mmol) were introduced into a 25 mL round-bottom flask degassed with nitrogen. A mixture of 1,4-dioxane and water (5 mL, 4:1) was added, and then stirred for 5 min, followed by the addition of Pd(PPh3)4 (4 mg, 0.003 mmol). The mixture was refluxed under a nitrogen atmosphere for 6 h. The organic phase was evaporated and the resulting oil was purified by column chromatography to obtain the coupling product 1-9.

[0190] General Scheme F: Reductive acylation. Compound 1-9 (0.56 mmol) and iron powder (158 mg, 2.82 mmol) were added to a reflux solution of methanol (8 mL), glacial acetic acid (1 mL) and water (1 mL) in sequence. The mixture was refluxed for another 30 min under vigorous stirring. The reaction solution was filtered through diatomaceous earth. The organic phase of the filtrate was removed in vacuo and a saturated K2CO3 aqueous solution (20 mL) was added to the residue. The resulting mixture was extracted with ethyl acetate, and the organic phase was washed with H2O, dried over Na2SO4, and concentrated to obtain the crude amino product amine 1-10, which was used directly in the next reaction step. All solid products obtained were dissolved in anhydrous THF solution, followed by trimethylamine (571 mg, 5.64 mmol). The mixture was cooled to 0°C and acyl chloride (2.82 mmol) was added dropwise. After stirring for 1 h at 0°C, the temperature was raised to room temperature and stirring was continued overnight. After removing the solvent in vacuo, the residue was purified by column chromatography to give amide 1-11 (152 mg, 66%).

[0191] General Scheme G: Oxidation reaction. To a stirred solution of 1-11 (0.028 mmol) in a mixture of CH3CN HO (1 mL, 11) was added dropwise a solution of cerium ammonium nitrate (31 mg, 0.056 mmol) in CH3CN: HO (1 mL, 2: 1) at 0°C. The reaction mixture was stirred at room temperature for 30 min, then diluted with ice water slurry (5 mL) and taken up by dichloromethane. The organic layers were combined, dried, concentrated and purified by column chromatography to give the oxidation product 1-12.

[0192] General Scheme H: Substitution reaction. To a solution of 1-12 (0.05 mmol) dissolved in dry chloroform (2 mL) was added amine (2.0 mmol) and the resulting red-brown solution was stirred at 0°C to room temperature until the starting material disappeared completely (TLC analysis). The resulting solution was evaporated under reduced pressure and the resulting crude material was purified by HPLC to afford pure product 1-c.

[0193] General Scheme I: Disubstitution reaction. To a solution of 1-12 (0.05 mmol) dissolved in dry chloroform (2 mL) was added amine (2.0 mmol, 200.3 mg) and the resulting reddish brown solution was stirred under reflux until the starting material completely disappeared (TLC analysis). The resulting solution was evaporated under reduced pressure and the resulting crude material was purified by HPLC to obtain the pure product (1-d).

[0194] General Scheme J: A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium (III) chloride heptahydrate (CeCl3 · 7H2O, 0.11 eq.) and an appropriate aromatic amine (0.1 eq.) in ethanol (2 mL) was stirred at 60-90°C for 2-6 h. Next, most of the ethanol was removed under vacuum to obtain a crude product, 6-arylamino-7-chloro-5,8-quinolinedione I. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were added to the above reaction system. The mixture solution was stirred at 65-90°C for 2-6 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to obtain compound II.

[0195] General Protocol N: Cytotoxicity.

[0196] MTT assay

[0197] The assessment of cytotoxicity is based on the reduction of MTT dye by living cells, producing a purple formazan product, which can be measured spectrophotometrically at 540 nm. 180 microliters of cancer cells were seeded into 96-well plates at 3,500-4,000 cells / well and incubated overnight at 37°C before designated treatment. After 72 h, 20 μl of MTT solution (3 mg / ml) was added and incubated again for 3 h. After removing the culture medium and dissolving the formazan crystals in 150 μL of DMSO, the absorbance was measured at 570 nm. The percentage of cell growth inhibition is expressed as 1-[(AB) / (CB)]x100% (A, B and C are the absorbance values ​​from experimental, blank and control cells, respectively).

[0198] Representative compounds were tested in the National Cancer Institute, Developmental Therapeutics Program 60 cell lines (NCI60).

[0199] Example II.

[0200] The compounds of formula I, formula II and formula III prepared by the above procedures 1 The HNMR data, LC / MS data and purity are summarized below:

[0201] Example 1: 7-Chloro-6-((2,6-difluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)quinoline-5,8-dione (1). General Protocol B was followed. A solution of 2,6-difluoro-4-(4-methylpiperazin-1-yl)aniline (22.7 mg, 0.1 mmol), 6,7-dichloroquinoline-5,8-dione (22.7 mg, 0.1 mmol) and cerium (III) chloride heptahydrate (41 mg, 0.11 mmol) in ethanol (2 mL) was stirred at 65 °C for 3 h. 7-Chloro-6-((2,6-difluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)quinoline-5,8-dione was recovered as a dark purple solid (28.8 mg, 69% yield). 1 H NMR (300 MHz, methanol-d4) δ9.01 (s, 1H), 8.55 (d, J = 7.7 Hz, 1H), 7.89 (s, 1H), 6.75 (d, J = 10.8 Hz, 2H), 3.97 (s, 2H), 3.64 (s, 2H), 3.35 (d, J = 11.5 Hz, 4H), 3.00 (s, 3H). LCMS (ESI) 419.00 [M+H] + . HPLC purity at 254 nm, 100%.

[0202] Example 2: 7-Chloro-6-((4-(4-(methylsulfonyl)piperazin-1-yl)phenyl)amino)quinoline-5,8-dione (2). General Protocol B was followed. A solution of 4-(4-(methylsulfonyl)piperazin-1-yl)aniline (25.5 mg, 0.1 mmol), 6,7-dichloroquinoline-5,8-dione (22.7 mg, 0.1 mmol) and cerium(III) chloride heptahydrate (41 mg, 0.11 mmol) in ethanol (2 mL) was stirred at 65° C. for 3 h. 7-Chloro-6-((4-(4-(methylsulfonyl)piperazin-1-yl)phenyl)amino)quinoline-5,8-dione was recovered as a dark purple solid (31.2 mg, 70% yield). 1 H NMR (300 MHz, methanol-d4) δ 8.91 (dd, J = 4.8, 1.7 Hz, 1H), 8.47 (dd, J = 7.9, 1.7 Hz, 1H), 7.76 (dd, J = 7.9, 4.8 Hz, 1H), 7.09 (d, J = 9.0 Hz, 2H), 6.98 (d, J = 9.0 Hz, 2H), 3.39-3.32 (m, 8H), 2.87 (s, 3H). LCMS (ESI) 447.00 [M+H] + HPLC purity at 254 nm, 95.6%.

[0203] Example 3: 7-Chloro-6-((4-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)phenyl)amino)quinoline-5,8-dione (3). General Protocol B was followed. A solution of 4-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)aniline (25.9 mg, 0.1 mmol), 6,7-dichloroquinoline-5,8-dione (22.7 mg, 0.1 mmol) and cerium (III) chloride heptahydrate (41 mg, 0.11 mmol) in ethanol (2 mL) was stirred at 65 °C for 3 h. 7-Chloro-6-((4-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)phenyl)amino)quinoline-5,8-dione was recovered as a dark purple solid (18.5 mg, 41% yield). 1 H NMR (300MHz, methanol-d4) δ8.96(d,J=5.1Hz,1H),8.53(d,J=6.4Hz,1H),7.87–7.77(m,1H),7.59–7. 47(m,2H),7.42(dd,J=8.6,2.3Hz,1H),3.62(d,J=9.9Hz,2H),3.28–3.20(m,6H),3.01(s,3H). LCMS(ESI)451.10[M+H] + HPLC purity at 254 nm, 98.3%.

[0204] Example 4: 5-((7-Chloro-5,8-dioxyylidene-5,8-dihydroquinolin-6-yl)amino)-2-(4-methylpiperazin-1-yl)benzonitrile (4). General Protocol B was followed. A solution of 5-amino-2-(4-methylpiperazin-1-yl)benzonitrile (21.6 mg, 0.1 mmol), 6,7-dichloroquinoline-5,8-dione (22.7 mg, 0.1 mmol) and cerium (III) chloride heptahydrate (41 mg, 0.11 mmol) in ethanol (2 mL) was stirred at 65 °C for 3 h. 5-((7-Chloro-5,8-dioxyylidene-5,8-dihydroquinolin-6-yl)amino)-2-(4-methylpiperazin-1-yl)benzonitrile was recovered as a dark purple solid (22.0 mg, 54% yield). 1 HNMR (300MHz, methanol-d4) δ8.97(d,J=4.4Hz,1H),8.52(d,J=7.8Hz,1H),7.82(dd,J=7.8,4.7Hz,1H),7.53(d,J=2.3Hz,1H),7.44(dd ,J=8.7,2.3Hz,1H),7.27(d,J=8.8Hz,1H),3.72(t,J=12.5Hz,4H),3.41(t,J=11.5Hz,2H),3.24(d,J=11.0Hz,2H),3.04(s,3H). LCMS(ESI)408.10[M+H] + HPLC purity at 254 nm, 99.2%.

[0205] Example 5: 7-Chloro-6-((4-(4-methylpiperazin-1-yl)phenyl)amino)quinoline-5,8-dione (5). General Protocol B was followed. A solution of 4-(4-methylpiperazin-1-yl)aniline (19.1 mg, 0.1 mmol), 6,7-dichloroquinoline-5,8-dione (22.7 mg, 0.1 mmol) and cerium (III) chloride heptahydrate (41 mg, 0.11 mmol) in ethanol (2 mL) was stirred at 65 °C for 3 h. 7-Chloro-6-((4-(4-methylpiperazin-1-yl)phenyl)amino)quinoline-5,8-dione was recovered as a dark purple solid (27.1 mg, 71% yield). 1H NMR (400 MHz, methanol-d4) δ 8.92 (s, 1H), 8.51 (dd, J = 7.9, 1.1 Hz, 1H), 7.79 (dd, J = 7.8, 4.8 Hz, 1H), 7.21-7.12 (m, 2H), 7.09-6.99 (m, 2H), 3.90 (d, J = 13.1 Hz, 2H), 3.70-3.60 (m, 2H), 3.36-3.26 (m, 2H), 3.09 (t, J = 12.1 Hz, 2H), 3.01 (s, 3H). LCMS (ESI) 383.1 [M+H] + . HPLC purity at 254 nm, 98.8%.

[0206] Example 6: 7-Chloro-6-((4-(piperidin-1-yl)phenyl)amino)quinoline-5,8-dione (6). General Protocol B was followed. A solution of 4-(piperidin-1-yl)aniline (17.6 mg, 0.1 mmol), 6,7-dichloroquinoline-5,8-dione (22.7 mg, 0.1 mmol) and cerium (III) chloride heptahydrate (41 mg, 0.11 mmol) in ethanol (2 mL) was stirred at 65 °C for 3 h. 7-Chloro-6-((4-(piperidin-1-yl)phenyl)amino)quinoline-5,8-dione was recovered as a dark purple solid (22.4 mg, 61% yield). 1 H NMR (300MHz, methanol-d4) δ8.97(d,J=4.4Hz,1H),8.53(d,J=7.6Hz,1H),7.83(dd,J=7.2,4.6Hz,1H),7.6 3(d,J=8.1Hz,2H),7.34(d,J=8.6Hz,2H),3.71–3.61(m,4H),2.13–1.99(m,4H),1.89–1.72(m,2H). LCMS(ESI)368.00[M+H] + HPLC purity at 254 nm, 98.9%.

[0207] Example 7: 6-((4-(1H-imidazol-1-yl)phenyl)amino)-7-chloroquinoline-5,8-dione (7). General Protocol B was followed. A solution of 4-(1H-imidazol-1-yl)aniline (15.9 mg, 0.1 mmol), 6,7-dichloroquinoline-5,8-dione (22.7 mg, 0.1 mmol) and cerium (III) chloride heptahydrate (41 mg, 0.11 mmol) in ethanol (2 mL) was stirred at 65 °C for 3 h. 6-((4-(1H-imidazol-1-yl)phenyl)amino)-7-chloroquinoline-5,8-dione was recovered as a dark purple solid (19.2 mg, 55% yield). 1H NMR (300MHz, methanol-d4) δ9.43(s,1H),8.98(dd,J=4.8,1.6Hz,1H),8.55(dd,J=7.9,1.7Hz,1H),8.09(t,J=1 .7Hz,1H),7.84(dd,J=7.9,4.8Hz,1H),7.79–7.76(m,1H),7.73(d,J=8.9Hz,2H),7.41(d,J=8.9Hz,2H). LCMS(ESI)351.00[M+H] + HPLC purity at 254 nm, 96.3%.

[0208] Example 8: 7-Chloro-6-((4-(pyridin-4-yl)phenyl)amino)quinoline-5,8-dione (8). General Protocol B was followed. A solution of 4-(pyridin-4-yl)aniline (17.0 mg, 0.1 mmol), 6,7-dichloroquinoline-5,8-dione (22.7 mg, 0.1 mmol) and cerium (III) chloride heptahydrate (41 mg, 0.11 mmol) in ethanol (2 mL) was stirred at 65 °C for 3 h. 7-Chloro-6-((4-(pyridin-4-yl)phenyl)amino)quinoline-5,8-dione was recovered as a dark purple solid (17.7 mg, 49% yield). 1 H NMR (300MHz, methanol-d4) δ8.99(dd,J=4.8,1.6Hz,1H),8.82(d,J=7.0Hz,2H),8.56(dd,J=7.9,1.7Hz, 1H), 8.40 (d, J = 7.0Hz, 2H), 8.07–8.00 (m, 2H), 7.85 (dd, J = 7.9, 4.8Hz, 1H), 7.37 (d, J = 8.8Hz, 2H). LCMS(ESI)361.90[M+H] + HPLC purity at 254 nm, 96.0%.

[0209] Example 9: 7-Chloro-6-((4-(thiazol-2-yl)phenyl)amino)quinoline-5,8-dione (9). General Protocol B was followed. A solution of 4-(thiazol-2-yl)aniline (17.6 mg, 0.1 mmol), 6,7-dichloroquinoline-5,8-dione (22.7 mg, 0.1 mmol) and cerium (III) chloride heptahydrate (41 mg, 0.11 mmol) in ethanol (2 mL) was stirred at 65 °C for 3 h. 7-Chloro-6-((4-(thiazol-2-yl)phenyl)amino)quinoline-5,8-dione was recovered as a dark purple solid (18.0 mg, 49% yield). 1H NMR (300MHz, methanol-d4) δ8.98(d,J=4.8Hz,1H),8.55(d,J=8.7Hz,1H),7.96(d,J=8.4Hz,2H),7.8 9 (d, J = 3.7 Hz, 1H), 7.84 (dd, J = 8.5, 4.2 Hz, 1H), 7.63 (d, J = 2.9 Hz, 1H), 7.27 (d, J = 8.6 Hz, 2H). LCMS(ESI)368.00[M+H] + HPLC purity at 254 nm, 97.9%.

[0210] Example 10: 6-((4-(4-acetylpiperazin-1-yl)phenyl)amino)-7-chloroquinoline-5,8-dione (10). General Protocol B was followed. A solution of 1-(4-(4-aminophenyl)piperazin-1-yl)ethan-1-one (21.9 mg, 0.1 mmol), 6,7-dichloroquinoline-5,8-dione (22.7 mg, 0.1 mmol) and cerium (III) chloride heptahydrate (41 mg, 0.11 mmol) in ethanol (2 mL) was stirred at 65 °C for 3 h. 6-((4-(4-acetylpiperazin-1-yl)phenyl)amino)-7-chloroquinoline-5,8-dione was recovered as a dark purple solid (25.4 mg, 62% yield). 1 H NMR (300MHz, methanol-d4) δ8.95(s,1H),8.52(d,J=7.6Hz,1H),7.85–7.76(m,1H),7.15(d,J =8.9Hz,2H),7.08(d,J=8.9Hz,2H),3.84–3.71(m,4H),3.29–3.23(m,4H),2.18(s,3H). LCMS(ESI)411.10[M+H] + HPLC purity at 254 nm, 99.4%.

[0211] Example 11: 7-Chloro-6-((3-fluoro-4-(piperazin-1-yl)phenyl)amino)quinoline-5,8-dione (11). General Protocol B was followed. A solution of tert-butyl 4-(4-amino-2-fluorophenyl)piperazine-1-carboxylate (29.5 mg, 0.1 mmol), 6,7-dichloroquinoline-5,8-dione (22.7 mg, 0.1 mmol) and cerium (III) chloride heptahydrate (41 mg, 0.11 mmol) in ethanol (2 mL) was stirred at 65 °C for 3 h. 7-Chloro-6-((3-fluoro-4-(piperazin-1-yl)phenyl)amino)quinoline-5,8-dione was recovered as a dark purple solid (23.5 mg, 61% yield). 1H NMR (300MHz, methanol-d4) δ8.96(d,J=4.8Hz,1H),8.52(dd,J=7.9,1.5Hz,1H),7.81(dd,J=7.8,4 .7Hz,1H),7.10(t,J=9.1Hz,1H),7.05–6.94(m,2H),3.46–3.40(m,4H),3.38–3.34(m,4H). LCMS(ESI)378.00[M+H] + HPLC purity at 254 nm, 99.5%.

[0212] Example 12: 7-Chloro-6-((4-(4-methoxyphenyl)piperazin-1-yl)phenyl)amino)quinoline-5,8-dione (12). General Protocol B was followed. A solution of 4-(4-(4-methoxyphenyl)piperazin-1-yl)aniline (28.3 mg, 0.1 mmol), 6,7-dichloroquinoline-5,8-dione (22.7 mg, 0.1 mmol) and cerium (III) chloride heptahydrate (41 mg, 0.11 mmol) in ethanol (2 mL) was stirred at 65 °C for 3 h. 7-Chloro-6-((4-(4-(4-methoxyphenyl)piperazin-1-yl)phenyl)amino)quinoline-5,8-dione was recovered as a dark purple solid (28.4 mg, 60% yield). 1 H NMR (300MHz, methanol-d4) δ8.96(d,J=5.5Hz,1H),8.52(d,J=7.4Hz,1H),7.85–7.77(m,1H),7.55( d,J=8.6Hz,2H),7.14(t,J=10.1Hz,6H),3.87(s,3H),3.80–3.74(m,4H),3.67–3.60(m,4H). LCMS(ESI)475.10[M+H] + HPLC purity at 254 nm, 98.9%.

[0213] Example 13: 7-Chloro-2-methyl-6-((4-(4-methylpiperazin-1-yl)phenyl)amino)quinoline-5,8-dione (13). General Protocol B was followed. A solution of 4-(4-methylpiperazin-1-yl)aniline (19.1 mg, 0.1 mmol), 6,7-dichloro-2-methylquinoline-5,8-dione (24.1 mg, 0.1 mmol) and cerium (III) chloride heptahydrate (41 mg, 0.11 mmol) in ethanol (2 mL) was stirred at 65 °C for 3 h. 7-Chloro-2-methyl-6-((4-(4-methylpiperazin-1-yl)phenyl)amino)quinoline-5,8-dione was recovered as a dark purple solid (23.4 mg, 59% yield). 1H NMR (300 MHz, methanol-d4) δ 8.37 (d, J = 8.0 Hz, 1H), 7.65 (d, J = 8.0 Hz, 1H), 7.17-7.12 (m, 2H), 7.04 (d, J = 9.1 Hz, 2H), 4.97 (s, 2H), 3.88 (s, 2H), 3.59 (d, J = 20.2 Hz, 2H), 3.08 (s, 2H), 3.00 (s, 3H), 2.72 (s, 3H). LCMS (ESI) 397.00 [M+H] + HPLC purity at 254 nm, 99.3%.

[0214] Example 14: 7-Chloro-6-((2,6-difluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)-2-methylquinoline-5,8-dione (14). General Protocol B was followed. A solution of 2,6-difluoro-4-(4-methylpiperazin-1-yl)aniline (22.7 mg, 0.1 mmol), 6,7-dichloro-2-methylquinoline-5,8-dione (24.1 mg, 0.1 mmol) and cerium (III) chloride heptahydrate (41 mg, 0.11 mmol) in ethanol (2 mL) was stirred at 65 °C for 3 h. 7-Chloro-6-((2,6-difluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)-2-methylquinoline-5,8-dione was recovered as a dark purple solid (30.2 mg, 70% yield). 1 HNMR (300 MHz, methanol-d4) δ8.36 (d, J = 8.1 Hz, 1H), 7.66 (d, J = 8.2 Hz, 1H), 6.76 (d, J = 10.2 Hz, 2H), 3.96 (s, 2H), 3.60 (s, 2H), 3.18 (dd, J = 35.3, 14.7 Hz, 4H), 3.00 (s, 3H), 2.72 (s, 3H). LCMS (ESI) 433.20 [M+H] + HPLC purity at 254 nm, 99.4%.

[0215] Example 15: 7-Chloro-2-methyl-6-((4-(4-(methylsulfonyl)piperazin-1-yl)phenyl)amino)quinoline-5,8-dione (15). General Protocol B was followed. A solution of 4-(4-(methylsulfonyl)piperazin-1-yl)aniline (25.5 mg, 0.1 mmol), 6,7-dichloro-2-methylquinoline-5,8-dione (24.1 mg, 0.1 mmol) and cerium (III) chloride heptahydrate (41 mg, 0.11 mmol) in ethanol (2 mL) was stirred at 65 °C for 3 h. 7-Chloro-2-methyl-6-((4-(4-(methylsulfonyl)piperazin-1-yl)phenyl)amino)quinoline-5,8-dione was recovered as a dark purple solid (35.4 mg, 77% yield). 1 H NMR (300 MHz, methanol-d4) δ 8.37 (d, J = 7.5 Hz, 1H), 7.65 (d, J = 8.7 Hz, 1H), 7.11 (d, J = 8.4 Hz, 2H), 7.02 (d, J = 8.8 Hz, 2H), 3.40 (d, J = 8.8 Hz, 8H), 2.91 (s, 3H), 2.72 (s, 3H). LCMS (ESI) 461.10 [M+H] + . HPLC purity at 254 nm, 98.8%.

[0216] Example 16: 5-(7-Chloro-2-methyl-5,8-dioxyylidene-5,8-dihydroquinolin-6-yl)-2-(4-methylpiperazin-1-yl)benzonitrile (16). General Protocol B was followed. A solution of 5-amino-2-(4-methylpiperazin-1-yl)benzonitrile (21.6 mg, 0.1 mmol), 6,7-dichloro-2-methylquinoline-5,8-dione (21.6 mg, 0.1 mmol) and cerium (III) chloride heptahydrate (41 mg, 0.11 mmol) in ethanol (2 mL) was stirred at 65 °C for 3 h. 5-((7-Chloro-2-methyl-5,8-dioxyylidene-5,8-dihydroquinolin-6-yl)amino)-2-(4-methylpiperazin-1-yl)benzonitrile was recovered as a dark purple solid (28.6 mg, 68% yield). 1 H NMR (300MHz, methanol-d4) δ8.35(d,J=7.7Hz,1H),7.65(d,J=7.6Hz,1H),7.50(s,1H),7.41(d,J=8.5Hz ,1H),7.24(d,J=8.8Hz,1H),3.70(t,J=14.1Hz,6H),3.45–3.36(m,2H),3.01(s,3H),2.71(s,3H). LCMS(ESI)422.10[M+H] + HPLC purity at 254 nm, 99.2%.

[0217] Example 17: 7-Chloro-2-methyl-6-((4-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)phenyl)amino)quinoline-5,8-dione (17). General Procedure B was followed. A solution of 4-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)aniline (25.9 mg, 0.1 mmol), 6,7-dichloro-2-methylquinoline-5,8-dione (24.1 mg, 0.1 mmol) and cerium (III) chloride heptahydrate (41 mg, 0.11 mmol) in ethanol (2 mL) was stirred at 65 °C for 3 h. 7-Chloro-2-methyl-6-((4-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)phenyl)amino)quinoline-5,8-dione was recovered as a dark purple solid (31.0 mg, 67% yield). 1 H NMR (300MHz, methanol-d4) δ8.38(d,J=8.3Hz,1H),7.68(d,J=8.3Hz,1H),7.56(d,J=8.9Hz,1H),7.51 (s,1H),7.41(d,J=6.8Hz,1H),3.62(d,J=10.7Hz,2H),3.25(s,6H),3.01(s,3H),2.73(s,3H). LCMS(ESI)456.20[M+H] + HPLC purity at 254 nm, 97.9%.

[0218] Example 18: 7-Chloro-6-[2-methoxy-4-(4-methylpiperazin-1-yl)phenyl]-2-methylquinoline-5,8-dione (18). General Protocol B was followed. A solution of 2-methoxy-4-(4-methylpiperazin-1-yl)aniline (22.1 mg, 0.1 mmol), 6,7-dichloro-2-methylquinoline-5,8-dione (24.1 mg, 0.1 mmol) and cerium (III) chloride heptahydrate (41 mg, 0.11 mmol) in ethanol (2 mL) was stirred at 65 °C for 3 h. 7-Chloro-6-((2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)-2-methylquinoline-5,8-dione was recovered as a dark purple solid (32.0 mg, 75% yield). 1H NMR (300 MHz, methanol-d4) δ 8.31 (d, J = 7.8 Hz, 1H), 7.61 (d, J = 8.0 Hz, 1H), 7.08 (d, J = 8.0 Hz, 1H), 6.65 (s, 1H), 6.59 (d, J = 9.2 Hz, 1H), 3.90 (s, 2H), 3.79 (s, 3H), 3.62 (s, 2H), 3.28-3.21 (m, 2H), 3.12 (d, J = 12.4 Hz, 2H), 2.99 (s, 3H), 2.70 (s, 3H). LCMS (ESI) 427.10 [M+H] + . HPLC purity at 254 nm, 98.8%.

[0219] Example 19: 7-Chloro-6-[3,5-difluoro-4-(4-methylpiperazin-1-yl)phenyl]quinoline-5,8-dione (19). General Protocol B was followed. A solution of 3,5-difluoro-4-(4-methylpiperazin-1-yl)aniline (22.7 mg, 0.1 mmol), 6,7-dichloroquinoline-5,8-dione (22.7 mg, 0.1 mmol) and cerium (III) chloride heptahydrate (41 mg, 0.11 mmol) in ethanol (2 mL) was stirred at 65 °C for 3 h. 7-Chloro-6-((3,5-difluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)quinoline-5,8-dione was recovered as a dark purple solid (33.0 mg, 79% yield). 1 H NMR (400MHz, DMSO-d6) δ9.78(brs,1H),9.39(s,1H),9.00(dd,J=4.7,1.7Hz,1H),8.39(dd,J=7.9 ,1.7Hz,1H),7.81(dd,J=7.9,4.7Hz,1H),6.88(d,J=11.0Hz,2H),3.37–3.10(m,8H),2.87(s,3H). LCMS(ESI)419.00[M+H] + HPLC purity at 254 nm, 98.2%.

[0220] Example 22: 7-chloro-6-[4-(ethylamino)-2-(trifluoromethyl)phenyl]quinoline-5,8-dione (22). According to general procedure B, N 1A solution of 7-ethyl-3-(trifluoromethyl)benzene-1,4-diamine (20.4 mg, 0.1 mmol), 6,7-dichloroquinoline-5,8-dione (22.7 mg, 0.1 mmol) and cerium(III) chloride heptahydrate (41 mg, 0.11 mmol) in ethanol (2 mL) was stirred at 65° C. for 3 h. 7-Chloro-6-((4-(ethylamino)-2-(trifluoromethyl)phenyl)amino)quinoline-5,8-dione was recovered as a dark purple solid (17.8 mg, 45% yield). 1 H NMR (400 MHz, methanol-d4) δ 8.93 (s, 1H), 8.45 (dd, J = 7.9, 1.6 Hz, 1H), 7.81 (dd, J = 7.9, 4.6 Hz, 1H), 7.16-7.07 (m, 2H), 6.89 (dt, J = 8.7, 0.7 Hz, 1H), 4.05-3.94 (m, 2H), 1.34-1.29 (m, 3H). LCMS (ESI) 396.00 [M+H] + HPLC purity at 254 nm, 97.4%.

[0221] Example 28: 7-Chloro-6-[2-fluoro-6-hydroxy-4-(4-methylpiperazin-1-yl)phenyl]quinoline-5,8-dione (28). General Scheme C was followed. 7-Chloro-6-((2,6-difluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)quinoline-5,8-dione 1 (0.66 mmol) was added to a stirred solution of KOH (0.37 g, 6.6 mmol) in MeOH:H2O (3:1) and heated to 80°C. After 2 h, the deep red solution was cooled to room temperature. 10 mL of ethyl acetate was added, washed with water (10 mL) and saturated aqueous NaCl solution (10 mL). The organic layer was dried over sodium sulfate, and the solvent was then distilled off under reduced pressure. It was purified by silica gel column chromatography to give the desired compound 28 (71%). 1 H NMR (400 MHz, methanol-d4) δ 8.73 (s, 1H), 8.46 (t, J = 8.2 Hz, 1H), 7.73 (s, 1H), 6.77-6.51 (m, 2H), 3.86 (s, 2H), 3.62 (s, 2H), 3.37-3.32 (m, 2H), 3.07 (s, 2H), 3.00 (s, 3H). LCMS (ESI) 417.00 [M+H] + . HPLC purity at 254 nm, 100%.

[0222] Example 31: (E)-4-chloro-N-(4-(4-fluorophenylvinyl)-5,8-dioxyylidene-7-(pyrrolidin-1-yl)-5,8-dihydroquinolin-6-yl)butanamide (31). General protocol H was followed. To a solution of 1-12 (0.05 mmol, 20 mg) dissolved in dry chloroform (2 mL) was added pyrrolidine (2.0 mmol, 142 mg) and the resulting red-brown solution was stirred at room temperature until the starting material disappeared completely (0.5 h, TLC analysis). The resulting solution was evaporated under reduced pressure and the resulting crude material was purified by HPLC to give the pure product (E)-4-chloro-N-(4-(4-fluorophenylvinyl)-5,8-dioxyylidene-7-(pyrrolidin-1-yl)-5,8-dihydroquinolin-6-yl)butanamide as a dark red solid (13.8 mg, 0.03 mmol, 59%), (HPLC purity at 254 nm, 98.2%). 1 H NMR (300MHz, CDCl3-d) δ8.78(d,J=4.8Hz,1H),8.45(d,J=16.4Hz,1H),7.75(d,J=8.6Hz,1H),7.64–7.59(m,2H ),7.49(s,1H),7.22–7.09(m,3H),3.77–3.58(m,6H),2.74–2.66(m,2H),2.25–2.14(m,2H),2.01–1.91(m,4H). LCMS(ESI)468.14[M+H] + .

[0223] Example 32: (E)-4-chloro-N-(4-(4-fluorophenylvinyl)-5,8-dioxy-7-(piperidin-1-yl)-5,8-dihydroquinolin-6-yl)butanamide (32). General protocol H was followed. To a solution of 1-12 (0.05 mmol, 20 mg) dissolved in dry chloroform (2 mL) was added piperidine (2.0 mmol, 170 mg) and the resulting red-brown solution was stirred at room temperature until the starting material completely disappeared (1 h, TLC analysis). The resulting solution was evaporated under reduced pressure and the resulting crude material was purified by HPLC to give the pure product (E)-4-chloro-N-(4-(4-fluorophenylvinyl)-5,8-dioxy-7-(piperidin-1-yl)-5,8-dihydroquinolin-6-yl)butanamide as a dark red solid (18.0 mg, 0.04 mmol, 75%), (HPLC purity at 254 nm, 97.0%). 1HNMR(300MHz, CDCl3-d)δ8.88(d,J=4.8Hz,1H),8.39(d,J=17.0Hz,1H),7.80(d,J=8.6Hz,1H),7.66–7.59(m,3H),7.49(s,1H) ,7.22–7.10(m,3H),3.69(t,J=6.2Hz,3H),3.48–3.40(m,4H),2.72(t,J=6.2Hz,2H),2.21(t,J=6.0Hz,3H),1.80–1.66(m,6H). LCMS(ESI)482.16[M+H] + .

[0224] Example 33: (E)-4-chloro-N-(4-(4-fluorophenylvinyl)-7-morpholine-5,8-dioxyylidene-5,8-dihydroquinolin-6-yl)butanamide (33). General protocol H was followed. To a solution of 1-12 (0.05 mmol, 20 mg) dissolved in dry chloroform (4 mL) was added morpholine (2.0 mmol, 170 mg) and the resulting red-brown solution was stirred at 50 °C until the starting material completely disappeared (8 h, TLC analysis). The resulting solution was evaporated under reduced pressure and the resulting crude material was purified by HPLC to give the pure product (E)-4-chloro-N-(4-(4-fluorophenylvinyl)-5,8-dioxyylidene-7-(piperidin-1-yl)-5,8-dihydroquinolin-6-yl)butanamide as a dark red solid (9.4 mg, 0.02 mmol, 39%), (HPLC purity at 254 nm, 99.5%). 1 H NMR (400MHz, DMSO) δ9.43(s,1H),8.86(d,J=5.1Hz,1H),8.27(d,J=16.5Hz,1H),8.02(d,J=5.2Hz,1H),7.69(dd,J=8.8,5.6Hz,2H),7.50(d ,J=16.3Hz,1H),7.31(t,J=8.9Hz,2H),3.98(d,J=13.5Hz,2H),3.75–3.60(m,4H),3.22–3.03(m,4H),2.71–2.64(m,1H),2.38–2.30(m,1H). 1.99–1.92(m,2H). LCMS(ESI)484.14[M+H] + .

[0225] Example 34: (E)-4-chloro-N-(4-(4-fluorophenylvinyl)-7-(4-methylpiperazin-1-yl)-5,8-dioxyylidene-5,8-dihydroquinolin-6-yl)butanamide (34). General protocol H was followed. To a solution of 1-12 (0.05 mmol, 20 mg) dissolved in dry chloroform (2 mL) was added 1-methylpiperazine (2.0 mmol, 200.3 mg) and the resulting red-brown solution was stirred at room temperature until the starting material disappeared completely (4 h, TLC analysis). The resulting solution was evaporated under reduced pressure and the resulting crude material was purified by HPLC to give the pure product (E)-4-chloro-N-(4-(4-fluorophenylvinyl)-7-(4-methylpiperazin-1-yl)-5,8-dioxyylidene-5,8-dihydroquinolin-6-yl)butanamide as a dark red solid (12.9 mg, 0.026 mmol, 52%), (HPLC purity at 254 nm, 98.8%). 1 H NMR (300MHz, CDCl3-d) δ8.89(d,J=5.5Hz,1H),8.31(d,J=16.0Hz,1H),7.81(d,J=5.2Hz,1H),7.76(s,1H),7.69–7.58(m,2H),7.28 –7.12(m,3H),3.71(t,J=5.9Hz,4H),3.17–3.05(m,2H),2.89(s,3H),2.75(t,J=7.0Hz,2H),2.62–2.45(m,4H),2.27–2.16(m,2H). LCMS(ESI)497.17[M+H] + .

[0226] Example 35: (E)-4-chloro-N-(4-(4-fluorophenylvinyl)-5,8-dioxyylidene-7-(piperazin-1-yl)-5,8-dihydroquinolin-6-yl)butanamide (35). General protocol H was followed. To a solution of 1-12 (0.05 mmol, 20 mg) dissolved in dry chloroform (2 mL) was added 1-methylpiperazine (2.0 mmol, 200.3 mg) and the resulting red-brown solution was stirred at room temperature until the starting material disappeared completely (2 h, TLC analysis). The resulting solution was evaporated under reduced pressure and the resulting crude material was purified by HPLC to give the pure product (E)-4-chloro-N-(4-(4-fluorophenylvinyl)-5,8-dioxyylidene-7-(piperazin-1-yl)-5,8-dihydroquinolin-6-yl)butanamide as a dark red solid (18.3 mg, 0.038 mmol, 76%), (HPLC purity at 254 nm, 100%). 1H NMR (400MHz, DMSO) δ9.53(s,1H),8.88(d,J=5.2Hz,1H),8.23(d,J=16.4Hz,1H),8.04(d,J=5.2Hz,1H),7.69(dd,J=8.5,5.7Hz,2H),7.52 (d,J=16.3Hz,1H),7.32(t,J=8.8Hz,2H),3.74(t,J=6.6Hz,2H),3.47(d,J=5.1Hz,4H),3.23(s,4H),2.61(d,J=7.4Hz,2H),2.05(s,2H). LCMS(ESI)483.15[M+H] + .

[0227] Example 36: (E)-N-(4-(4-fluorophenylvinyl)-7-(4-methylpiperazin-1-yl)-5,8-dioxyylidene-5,8-dihydroquinolin-6-yl)-4-(4-methylpiperazin-1-yl)butanamide (36). General Protocol I was followed. To a solution of 1-12 (0.05 mmol, 20 mg) dissolved in dry chloroform (2 mL) was added 1-methylpiperazine (2.0 mmol, 200.3 mg) and the resulting red-brown solution was stirred at reflux overnight until the starting material disappeared completely (TLC analysis). The resulting solution was evaporated under reduced pressure and the resulting crude material was purified by HPLC to give the pure product (E)-N-(4-(4-fluorophenylvinyl)-7-(4-methylpiperazin-1-yl)-5,8-dioxyylidene-5,8-dihydroquinolin-6-yl)-4-(4-methylpiperazin-1-yl)butanamide as a dark red solid (17.6 mg, 63%), (HPLC purity at 254 nm, 100%). 1 H NMR (400 MHz, acetone) δ8.86 (d, J = 5.1 Hz, 1H), 8.36 (d, J = 16.4 Hz, 1H), 8.00 (d, J = 5.1 Hz, 1H), 7.74 (dd, J = 8.7, 5.5 Hz, 2H), 7.48 (d, J = 16.3 Hz, 1H), 7.25 (t, J = 8.8 Hz, 2H), 3.76 (s, 6H), 3.57–3.41 (m, 10H), 3.14 (dd, J = 10.0, 5.4 Hz, 2H), 2.98 (s, 3H), 2.85 (s, 3H), 2.77 (t, J = 6.9 Hz, 2H), 2.24–2.20 (m, 1H), 1.93–1.89 (m, 1H). or 1HNMR(400MHz,MeOD)δ8.82(d,J=5.2Hz,1H),8.37(d,J=16.4Hz,1H),8.06(d,J=5 .2Hz,1H),7.72(dd,J=8.2,5.6Hz,2H),7.47(d,J=16.4Hz,1H),7.20(t,J=8.6Hz, 2H),3.83(s,2H),3.53(d,J=30.2Hz,6H),3.15(s,6H),2.99(s,3H),2.95–2.88( m,2H),2.88–2.78(m,2H),2.82(s,3H),2.70(t,J=6.8Hz,2H),2.09–1.99(m,2H).

[0228] Example 37: (E)-N-(4-(4-fluorophenylvinyl)-5,8-dimethoxyquinolin-6-yl)pentanamide (37). General Scheme F was followed. Compound 1-9 (200 mg, 0.59 mmol) and iron powder (332 mg, 5.9 mmol) were added to a refluxing solution of methanol (8 mL), glacial acetic acid (1 mL) and water (1 mL) in sequence. The mixture was refluxed for another 30 min under vigorous stirring. The reaction solution was filtered through diatomaceous earth. The organic phase of the filtrate was removed in vacuo and a saturated aqueous K2CO3 solution (20 mL) was added to the residue. The resulting mixture was extracted with ethyl acetate, and the organic phase was washed with H2O, dried over Na2SO4, and concentrated to give the crude amino product 1-10, which was pure enough to proceed to the next reaction step. All the solid product obtained was dissolved in anhydrous dichloromethane solution, followed by trimethylamine (571 mg, 4.76 mmol). The mixture was cooled to 0°C and valeryl chloride (419 mg, 2.97 mmol) was added dropwise. After stirring at 0°C for 1 h, the temperature was raised to room temperature and stirring was continued overnight. After removing the solvent in vacuo, the residue was purified by column chromatography to give (E)-N-(4-(4-fluorophenylvinyl)-5,8-dimethoxyquinolin-6-yl)valeramide (180.5 mg, 75%) as a yellow solid.

[0229] 1 H NMR (300MHz, CDCl3) δ9.07(s,1H),8.54(s,1H),8.29(d,J=16.0Hz,2H),7.74(s,1H),7.67(dd,J=8.6,5. 4Hz,2H),7.32(d,J=16.2Hz,1H),7.19(t,J=8.5Hz,2H),4.09(s,3H),3.66(s,3H),2.57(t,J=7.5Hz,2H),

[0230] 1.86–1.73(m,2H), 1.49(dd,J=15.1,7.5Hz,2H), 1.01(t,J=7.3Hz,3H).

[0231] Example 38: (E)-N-(4-(4-fluorophenylvinyl)-5,8-dioxy-5,8-dihydroquinolin-6-yl)pentanamide (38). General Protocol G was followed. A solution of 37 (13 mg, 0.032 mmol) in (7:3) acetonitrile:water (1 mL) was cooled at 0°C in an ice bath, and a solution of ceric ammonium nitrate (2.7 eq, 52 mg, 0.09 mmol) in (9:1) acetonitrile:water (1 mL) was added dropwise. The reaction mixture was stirred for 15 min, then poured into ice / water and extracted with CHCl (5 times). The organic layer was washed with water (5 times), dried over anhydrous Na2SO4 and concentrated to dryness, and the crude material was purified by HPLC to give the pure product (E)-N-(4-(4-fluorophenylvinyl)-5,8-dioxy-5,8-dihydroquinolin-6-yl)pentanamide as a dark red solid (9.4 mg, 78%), (HPLC purity at 254 nm, 100%). 1 H NMR (300MHz, CDCl3) δ8.97(d,J=5.1Hz,1H),8.41(s,1H),8.27(d,J=16.1Hz,1H),8.05(s,1H),7.81(d,J=5.1Hz,1H),7. 69–7.60(m,2H),7.16(t,J=8.6Hz,2H),2.55–2.47(m,2H),1.79–1.70(m,2H),1.49–1.39(m,2H),0.98(t,J=7.3Hz,3H).

[0232] Example 39: (E)-N-(4-(4-fluorophenylvinyl)-5,8-dioxy-7-(pyrrolidin-1-yl)-5,8-dihydroquinolin-6-yl)pentanamide (39). General protocol H was followed. To a solution of 38 (0.05 mmol, 17 mg) dissolved in dry chloroform (2 mL) was added pyrrolidine (2.0 mmol, 142 mg) and the resulting red-brown solution was stirred at room temperature until the starting material completely disappeared (0.5 h, TLC analysis). The resulting solution was evaporated under reduced pressure and the resulting crude material was purified by HPLC to give the pure product (E)-N-(4-(4-fluorophenylvinyl)-5,8-dioxy-7-(pyrrolidin-1-yl)-5,8-dihydroquinolin-6-yl)pentanamide as a dark red solid (14.8 mg, 66%), (HPLC purity at 254 nm, 97.9%). 1H NMR (300MHz, CDCl3) δ8.76(d,J=5.2Hz,1H),8.44(d,J=16.4Hz,1H),7.72(s,1H),7.65(d,J=1.5Hz,1H),7.48(dd,J=3.0,1.5Hz,2H),7.20–7.06(m,3H) ,3.64(d,J=2.6Hz,4H),2.48(dd,J=13.7,5.9Hz,2H),1.99–1.86(m,4H),1. 72(dt,J=15.3,7.7Hz,2H), 1.42(dd,J=14.8,7.4Hz,2H), 1.03–0.91(m,3H).

[0233] Example 40: (E)-N-(4-(4-fluorophenylvinyl)-7-morpholino-5,8-dioxy-5,8-dihydroquinolin-6-yl)pentanamide (40). General protocol H was followed. To a solution of 38 (0.05 mmol, 17 mg) dissolved in dry chloroform (4 mL) was added morpholine (2.0 mmol, 170 mg) and the resulting red-brown solution was stirred at room temperature until the starting material completely disappeared (8 h, TLC analysis). The resulting solution was evaporated under reduced pressure and the resulting crude material was purified by HPLC to give the pure product (E)-N-(4-(4-fluorophenylvinyl)-7-morpholino-5,8-dioxy-5,8-dihydroquinolin-6-yl)pentanamide as a dark red solid (12.3 mg, 53%), (HPLC purity at 254 nm, 99%). 1 H NMR (300MHz, CDCl3) δ8.86(d,J=5.2Hz,1H),8.35(d,J=16.2Hz,1H),7.77(d,J=5.1Hz,1H),7.67–7.57(m,3H),7.21(d,J=16.2Hz,1H),7.13(t, J=8.6Hz,2H),3.91–3.81(m,4H),3.54–3.44(m,4H),2.56–2.45(m,2H),1.81–1.66(m,2H),1.44(dd,J=15.0,7.4Hz,2H),0.98(t,J=7.3Hz,3H).

[0234] Example 41: (E)-N-(4-(4-fluorophenylvinyl)-7-(4-methylpiperazin-1-yl)-5,8-dioxyylidene-5,8-dihydroquinolin-6-yl)pentanamide (41). General protocol H was followed. To a solution of 38 (0.05 mmol, 17 mg) dissolved in dry chloroform (2 mL) was added 1-methylpiperazine (2.0 mmol, 200.3 mg) and the resulting red-brown solution was stirred at room temperature until the starting material completely disappeared (4 h, TLC analysis). The resulting solution was evaporated under reduced pressure and the resulting crude material was purified by HPLC to give the pure product (E)-N-(4-(4-fluorophenylvinyl)-7-(4-methylpiperazin-1-yl)-5,8-dioxyylidene-5,8-dihydroquinolin-6-yl)pentanamide as a dark red solid (17.6 mg, 74%), (HPLC purity at 254 nm, 97.6%). 1 H NMR (300MHz, CDCl3) δ8.83(d,J=5.2Hz,1H),8.32(d,J=16.4Hz,1H),7.73(d,J=5.3Hz,1H),7.66–7.56(m,3H),7.23–7.08(m,3H),3.60 –3.51(m,4H),2.84(s,4H),2.52(s,3H),2.47(d,J=8.0Hz,2H),1.70(d,J=7.6Hz,2H),1.48–1.35(m,2H),0.95(dd,J=13.9,6.7Hz,3H).

[0235] Example 42: 9-(4-Methylpiperazin-1-yl)pyrido[2,3-b]phenazine-5,12-dione (42). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-(4-methylpiperazin-1-yl)aniline (19.1 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-(4-methylpiperazin-1-yl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (9.7 mg, 27% yield). 1H NMR (300MHz, methanol-d4) δ9.13 (d, J = 5.0 Hz, 1H), 8.85 (d, J = 8.4 Hz, 1H), 8.24 (d, J = 9. 5Hz,1H),8.11–7.94(m,2H),7.58(s,1H),3.91(s,4H),3.52(s,4H),3.02(s,3H). LCMS(ESI)360.00[M+H] + HPLC purity at 254 nm, 99.2%.

[0236] Example 43: 7-Methoxy-9-(4-methylpiperazin-1-yl)pyrido[2,3-b]phenazine-5,12-dione (43). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 2-methoxy-4-(4-methylpiperazin-1-yl)aniline (22.1 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 7-methoxy-9-(4-methylpiperazin-1-yl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (19.1 mg, 49% yield). 1 H NMR (300 MHz, methanol-d4) δ9.13 (s, 1H), 8.82 (d, J = 6.9 Hz, 1H), 8.03 (d, J = 7.6 Hz, 1H), 7.12 (s, 1H), 6.93 (s, 1H), 4.06 (s, 3H), 3.59 (s, 8H), 3.09 (s, 3H). LCMS (ESI) 390.10 [M+H] + HPLC purity at 254 nm, 99.3%.

[0237] Example 44: 2-Methyl-9-(4-methylpiperazin-1-yl)-5,12-dioxy-5,12-dihydropyrido[2,3-b]phenazine-10-carbonitrile (44). General Scheme J was followed. A solution of 6,7-dichloro-2-methylquinoline-5,8-dione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 5-amino-2-(4-methylpiperazin-1-yl)benzonitrile (21.6 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 2-methyl-9-(4-methylpiperazin-1-yl)-5,12-dioxy-5,12-dihydropyrido[2,3-b]phenazine-10-carbonitrile as a dark purple solid (19.9 mg, 50% yield). 1 HNMR (400MHz, methanol-d4) δ8.72 (d, J = 8.1 Hz, 1H), 8.36 (d, J = 9.5 Hz, 1H), 8.00 (d, J = 9.5 Hz, 1H), 7.89 (d, J = 8.2 Hz, 1H), 3.82-3.46 (m, 6H), 3.37 (s, 2H), 3.09 (s, 3H), 2.84 (s, 3H). LCMS (ESI) 399.05 [M+H] + . HPLC purity at 254 nm, 100%.

[0238] Example 45: 9-(4-Methylpiperazin-1-yl)-5,12-dioxy-5,12-dihydropyrido[2,3-b]phenazine-10-carbonitrile (45). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 5-amino-2-(4-methylpiperazin-1-yl)benzonitrile (21.6 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-(4-methylpiperazin-1-yl)-5,12-dioxy-5,12-dihydropyrido[2,3-b]phenazine-10-carbonitrile as a dark purple solid (11.1 mg, 29% yield). 1 H NMR(400MHz, DMSO-d6)δ9.17(dd,J=4.6,1.7Hz,1H),9.07(s,1H),8.70(dd,J=7.9,1.7 Hz, 1H), 8.06 (s, 1H), 8.00 (dd, J = 7.9, 4.6Hz, 1H), 4.00 (s, 6H), 2.90 (d, J = 17.4Hz, 5H). LCMS(ESI)385.25[M+H] + HPLC purity at 254 nm, 98.5%.

[0239] Example 46: 9-(4-Methylpiperazin-1-yl)-5,12-dioxy-5,12-dihydropyrido[2,3-b]phenazine-8-carbonitrile (46). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 5-amino-2-(4-methylpiperazin-1-yl)benzonitrile (21.6 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-(4-methylpiperazin-1-yl)-5,12-dioxy-5,12-dihydropyrido[2,3-b]phenazine-8-carbonitrile as a dark purple solid (8.4 mg, 22% yield). 1 HNMR(400MHz,DMSO-d6)δ9.17(dd,J=4.6,1.7Hz,1H),9.07(s,1H),8.70(dd,J=7.9,1.7 Hz, 1H), 8.06 (s, 1H), 8.00 (dd, J = 7.9, 4.6Hz, 1H), 4.00 (s, 6H), 2.90 (d, J = 17.4Hz, 5H). LCMS(ESI)384.95[M+H] + HPLC purity at 254 nm, 98.1%.

[0240] Example 47: 9-(4-(Methylsulfonyl)piperazin-1-yl)-5,12-dioxy-5,12-dihydropyrido[2,3-b]phenazine-10-carbonitrile (47). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 5-amino-2-(4-(methylsulfonyl)piperazin-1-yl)benzonitrile (28.0 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-(4-(methylsulfonyl)piperazin-1-yl)-5,12-dioxy-5,12-dihydropyrido[2,3-b]phenazine-10-carbonitrile as a dark purple solid (22.0 mg, 49% yield). 1 HNMR(400MHz,DMSO-d6)δ9.16(dd,J=4.6,1.6Hz,1H),8.67(dd,J=7.9,1.6Hz,1H),8.42(d,J=9.8H z, 1H), 8.03 (d, J = 9.5Hz, 1H), 7.98 (dd, J = 7.8, 4.6Hz, 1H), 3.98 (s, 4H), 3.42 (s, 4H), 3.00 (s, 3H). LCMS(ESI)448.90[M+H] + HPLC purity at 254 nm, 96.0%.

[0241] Example 48: 9-(4-Ethylpiperazin-1-yl)pyrido[2,3-b]phenazine-5,12-dione (48). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-(4-ethylpiperazin-1-yl)aniline (20.5 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-(4-ethylpiperazin-1-yl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (11.6 mg, 31% yield). 1 H NMR (400MHz, DMSO-d6) δ9.14(dd,J=4.6,1.7Hz,1H),8.66(dd,J=7.9,1.7Hz,1H),8.26(d,J=9.5Hz,1H),8.08(d,J=2.7Hz,1H ), 7.96 (dd, J = 7.9, 4.6Hz, 1H), 7.67 (d, J = 2.7Hz, 1H), 4.44 (s, 2H), 3.66 (s, 4H), 3.24 (q, J = 7.1Hz, 4H), 1.29 (t, J = 7.3Hz, 3H). LCMS(ESI)374.05[M+H] + . HPLC purity at 254 nm, 100%.

[0242] Example 49: 9-(4-Cyclopropylpiperazin-1-yl)pyrido[2,3-b]phenazine-5,12-dione (49). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-(4-cyclopropylpiperazin-1-yl)aniline (21.7 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-(4-cyclopropylpiperazin-1-yl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (23.5 mg, 61% yield). 1 HNMR (400MHz, methanol-d4) δ9.13(dd,J=4.6,1.6Hz,1H),8.85(dd,J=8.0,1.7Hz,1H),8.24(d,J=9.5Hz,1H),8.03(ddd ,J=12.6,8.8,3.7Hz,2H),7.58(d,J=2.7Hz,1H),3.94(s,4H),3.69(s,4H),3.04–2.90(m,1H),1.16–1.02(m,4H). LCMS(ESI)386.05[M+H] + . HPLC purity at 254 nm, 100%.

[0243] Example 50: 9-(4-Methylpiperazin-1-yl)-10-(trifluoromethyl)pyrido[2,3-b]phenazine-5,12-dione (50). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)aniline (25.9 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-(4-methylpiperazin-1-yl)-10-(trifluoromethyl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (14.5 mg, 34% yield). 1 H NMR (400MHz, methanol-d4) δ9.15 (dd, J=4.6, 1.5Hz, 1H), 8.85 (dd, J=8.0, 1.7Hz, 1H), 8.39 (d, J=9. 5Hz, 1H), 8.10 (d, J = 9.6Hz, 1H), 8.02 (dd, J = 8.0, 4.7Hz, 1H), 4.01–3.40 (m, 8H), 3.08 (s, 3H). LCMS(ESI)428.10[M+H] + HPLC purity at 254 nm, 99.1%.

[0244] Example 51: 9-(4-Methylpiperazin-1-yl)-8-(trifluoromethyl)pyrido[2,3-b]phenazine-5,12-dione (51). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)aniline (25.9 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 2-methyl-9-(4-methylpiperazin-1-yl)-8-(trifluoromethyl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (7.1 mg, 16% yield). 1 H NMR (400MHz, methanol-d4) δ9.18(dd,J=4.6,1.6Hz,1H),8.89(dd,J=8.0,1.7Hz,1H),8.80(s,1H),8.40(s,1H ), 8.05 (dd, J = 8.0, 4.7Hz, 1H), 3.380-3.71 (m, 2H), 3.65-3.57 (m, 2H), 3.50-3.39 (m, 4H), 3.08 (s, 3H). LCMS(ESI)428.10[M+H] + . HPLC purity at 254 nm, 99.8%.

[0245] Example 52: 2-Methyl-9-(4-methylpiperazin-1-yl)-10-(trifluoromethyl)pyrido[2,3-b]phenazine-5,12-dione (52). General Scheme J was followed. A solution of 6,7-dichloro-2-methylquinoline-5,8-dione (0.1 eq., 24 mg), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)aniline (25.9 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 2-methyl-9-(4-methylpiperazin-1-yl)-10-(trifluoromethyl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (11.0 mg, 25% yield). 1 H NMR (400 MHz, methanol-d4) δ 8.69 (d, J = 8.1 Hz, 1H), 8.39 (d, J = 9.5 Hz, 1H), 8.10 (d, J = 9.5 Hz, 1H), 7.87 (d, J = 8.1 Hz, 1H), 3.93 (s, 2H), 3.71 (s, 4H), 3.53-3.38 (m, 2H), 3.08 (s, 3H), 2.83 (s, 3H). LCMS (ESI) 442.10 [M+H] + HPLC purity at 254 nm, 95.8%.

[0246] Example 53: 2-Methyl-9-(4-methylpiperazin-1-yl)-8-(trifluoromethyl)pyrido[2,3-b]phenazine-5,12-dione (53). General Scheme J was followed. A solution of 6,7-dichloro-2-methylquinoline-5,8-dione (0.1 eq., 24 mg), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)aniline (25.9 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 2-methyl-9-(4-methylpiperazin-1-yl)-8-(trifluoromethyl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (7.1 mg, 16% yield). 1 H NMR (400 MHz, methanol-d4) δ 8.81 (s, 1H), 8.75 (d, J = 8.1 Hz, 1H), 8.40 (s, 1H), 7.91 (d, J = 8.2 Hz, 1H), 3.72 (s, 2H), 3.61 (s, 2H), 3.42 (d, J = 8.4 Hz, 4H), 3.08 (s, 3H), 2.85 (s, 3H). LCMS (ESI) 441.95 [M+H] + HPLC purity at 254 nm, 98.7%.

[0247] Example 54: 9-(4-Cyclopropylpiperazin-1-yl)-2-methylpyrido[2,3-b]phenazine-5,12-dione (54). General Scheme J was followed. A solution of 6,7-dichloro-2-methylquinoline-5,8-dione (0.1 eq., 24 mg), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-(4-cyclopropylpiperazin-1-yl)aniline (21.7 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-(4-cyclopropylpiperazin-1-yl)-2-methylpyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (21.9 mg, 55% yield). 1 H NMR (400MHz, methanol-d4) δ8.70(d,J=8.1Hz,1H),8.23(d,J=9.5Hz,1H),8.04(dd,J=9.6,2.8Hz,1H),7.87(d,J=8.2H z, 1H), 7.58 (d, J = 2.7Hz, 1H), 3.92 (s, 4H), 3.64 (s, 4H), 2.90 (s, 1H), 2.83 (s, 3H), 1.05 (dd, J = 11.1, 6.1Hz, 4H). LCMS(ESI)400.15[M+H] + HPLC purity at 254 nm, 99.4%.

[0248] Example 55: 10-Fluoro-9-(4-methylpiperazin-1-yl)pyrido[2,3-b]phenazine-5,12-dione (55). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 3-fluoro-4-(4-methylpiperazin-1-yl)aniline (20.9 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 10-fluoro-9-(4-methylpiperazin-1-yl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (7.9 mg, 21% yield). 1 H NMR (400 MHz, methanol-d4) δ 9.15 (dd, J = 4.8, 1.7 Hz, 1H), 8.87 (dd, J = 8.0, 1.7 Hz, 1H), 8.18 (dd, J = 9.5, 1.4 Hz, 1H), 8.10-7.96 (m, 2H), 4.15 (s, 2H), 3.81-3.44 (m, 6H), 3.07 (s, 3H). LCMS (ESI) 378.10 [M+H] + . HPLC purity at 254 nm, 100%.

[0249] Example 56: 10-Chloro-9-(4-methylpiperazin-1-yl)pyrido[2,3-b]phenazine-5,12-dione (56). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 3-chloro-4-(4-methylpiperazin-1-yl)aniline (22.5 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 10-chloro-9-(4-methylpiperazin-1-yl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (13.0 mg, 33% yield). 1H NMR (499MHz, methanol-d4) δ9.14(d,J=3.3Hz,1H),8.85(dd,J=7.9,1.6Hz,1H),8.30(d,J=9.3Hz,1H),8.0 4(d,J=9.3Hz,1H),8.01(dd,J=8.0,4.7Hz,1H),4.03(s,2H),3.72(s,2H),3.49(s,4H),3.06(s,3H). 19 F NMR (470 MHz, methanol-d4) δ-76.99. LCMS (ESI) 393.90 [M+H] + HPLC purity at 254 nm, 99.0%.

[0250] Example 57: 9-(4-(Methylsulfonyl)piperazin-1-yl)pyrido[2,3-b]phenazine-5,12-dione (57). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-(4-(methylsulfonyl)piperazin-1-yl)aniline (25.5 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-(4-(methylsulfonyl)piperazin-1-yl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (17.3 mg, 41% yield). 1 H NMR (300MHz, chloroform-d) δ9.24(dd,J=4.6,1.7Hz,1H),8.84(dd,J=8.0,1.8Hz,1H),8.34(d,J=9.6Hz,1H),7.87(dd,J=8.0 ,4.6Hz,1H),7.79(dd,J=9.6,2.9Hz,1H),7.63(d,J=3.0Hz,1H),3.85–3.68(m,4H),3.56–3.39(m,4H),2.89(s,3H). LCMS(ESI)424.10[M+H] + HPLC purity at 254 nm, 99.2%.

[0251] Example 58: 2-Methyl-9-(4-(methylsulfonyl)piperazin-1-yl)pyrido[2,3-b]phenazine-5,12-dione (58). General Scheme J was followed. A solution of 6,7-dichloro-2-methylquinoline-5,8-dione (0.1 eq., 24 mg), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-(4-(methylsulfonyl)piperazin-1-yl)aniline (25.5 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 2-methyl-9-(4-(methylsulfonyl)piperazin-1-yl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (20.1 mg, 46% yield). 1 HNMR (300MHz, chloroform-d) δ8.69 (d, J = 7.7 Hz, 1H), 8.32 (d, J = 9.5 Hz, 1H), 7.78 (s, 1H), 7.72-7.62 (m, 2H), 3.72 (s, 4H), 3.49 (s, 4H), 2.88 (s, 3H), 2.86 (s, 3H). LCMS (ESI) 438.05 [M+H] + . HPLC purity at 254 nm, 100%.

[0252] Example 59: 10-Chloro-2-methyl-9-(4-methylpiperazin-1-yl)pyrido[2,3-b]phenazine-5,12-dione (59). General Scheme J was followed. A solution of 6,7-dichloro-2-methylquinoline-5,8-dione (0.1 eq., 24 mg), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 3-chloro-4-(4-methylpiperazin-1-yl)aniline (22.5 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 10-chloro-2-methyl-9-(4-methylpiperazin-1-yl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (15.9 mg, 39% yield). 1H NMR (300MHz, DMSO-d6) δ8.57(d,J=8.0Hz,1H),8.35(d,J=9.2Hz,1H),8.09(d,J=9.4 Hz,1H),7.85(d,J=8.1Hz,1H),3.92(s,4H),3.37(s,4H),2.95(s,3H),2.76(s,3H). LCMS(ESI)408.05[M+H] + HPLC purity at 254 nm, 95.5%.

[0253] Example 60: 9-((4-ethylpiperazin-1-yl)methyl)-2-methylpyrido[2,3-b]phenazine-5,12-dione (60). General Scheme J was followed. A solution of 6,7-dichloro-2-methylquinoline-5,8-dione (0.1 eq., 24 mg), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-((4-ethylpiperazin-1-yl)methyl)aniline (21.9 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-((4-ethylpiperazin-1-yl)methyl)-2-methylpyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (9.6 mg, 24% yield). 1 HNMR (400MHz, methanol-d4) δ8.74 (d, J = 8.1 Hz, 1H), 8.42 (d, J = 8.6 Hz, 2H), 8.19 (dd, J = 8.9, 1.7 Hz, 1H), 7.90 (d, J = 8.1 Hz, 1H), 4.02 (s, 2H), 3.72-3.46 (m, 2H), 3.30-3.13 (m, 6H), 2.84 (s, 3H), 2.74-2.55 (m, 2H), 1.39 (t, J = 7.3 Hz, 3H). LCMS (ESI) 402.20 [M+H] + HPLC purity at 254 nm, 99.5%.

[0254] Example 61: 9-(4-Methylpiperazine-1-carbonyl)-8-(trifluoromethyl)pyrido[2,3-b]phenazine-5,12-dione (61). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and (4-amino-2-(trifluoromethyl)phenyl)(4-ethylpiperazin-1-yl)methanone (30.1 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-(4-methylpiperazine-1-carbonyl)-8-(trifluoromethyl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (10.0 mg, 22% yield). 1 HNMR (300MHz, methanol-d4) δ9.19 (dd, J = 4.6, 1.7 Hz, 1H), 8.99-8.85 (m, 2H), 8.64 (s, 1H), 8.06 (dd, J = 8.0, 4.7 Hz, 1H), 3.67 (d, J = 74.0 Hz, 8H), 3.03 (s, 3H). LCMS (ESI) 456.05 [M+H] + . HPLC purity at 254 nm, 98.8%.

[0255] Example 62: 9-Morpholino-10-(trifluoromethyl)pyrido[2,3-b]phenazine-5,12-dione (62). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium (III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-morpholino-3-(trifluoromethyl)aniline (24.6 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-morpholino-10-(trifluoromethyl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (12 mg, 29% yield). 1HNMR (300MHz, chloroform-d) δ9.25 (d, J=4.6Hz, 1H), 8.83 (dd, J=8.0, 1.7Hz, 1H), 8.42 (d,J=9.6Hz,1H),7.95–7.79(m,2H),4.01–3.89(m,4H),3.58(d,J=5.1Hz,4H). LCMS(ESI)415.00[M+H] + . HPLC purity at 254 nm, 99.7%.

[0256] Example 63: 9-Morpholino-8-(trifluoromethyl)pyrido[2,3-b]phenazine-5,12-dione (63). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium (III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-morpholino-3-(trifluoromethyl)aniline (24.6 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-morpholino-8-(trifluoromethyl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (7.9 mg, 19% yield). 1 HNMR (300 MHz, chloroform-d) δ9.30 (d, J = 2.7 Hz, 1H), 8.93–8.83 (m, 2H), 8.19 (s, 1H), 7.93 (dd, J = 8.0, 4.6 Hz, 1H), 4.05–3.90 (m, 4H), 3.34–3.20 (m, 4H). LCMS (ESI) 414.95 [M+H] + HPLC purity at 254 nm, 98.7%.

[0257] Example 64: 9-(4-Methylpiperidin-1-yl)pyrido[2,3-b]phenazine-5,12-dione (64). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-(4-methylpiperidin-1-yl)aniline (19.0 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-(4-methylpiperidin-1-yl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (19.3 mg, 54% yield). 1 H NMR (400MHz, methanol-d4) δ9.10(s,1H),8.82(d,J=7.7Hz,1H),8.13(d,J=9.6Hz,1H),8.00(d,J=9.3Hz,2H),7.43(d,J=2.7 Hz,1H),4.29(d,J=13.2Hz,2H),3.27–3.11(m,2H),1.90(d,J=13.6Hz,2H),1.45–1.27(m,3H),1.04(d,J=6.4Hz,3H). LCMS(ESI)359.00[M+H] + . HPLC purity at 254 nm, 100%.

[0258] Example 65: 9-(4-Methylpiperazin-1-yl)-7-(trifluoromethyl)pyrido[2,3-b]phenazine-5,12-dione (65). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-(4-methylpiperazin-1-yl)-2-(trifluoromethyl)aniline (25.8 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-(4-methylpiperazin-1-yl)-7-(trifluoromethyl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (20.9 mg, 49% yield). 1 H NMR (400 MHz, methanol-d4) δ 9.13 (dd, J = 4.6, 1.7 Hz, 1H), 8.90–8.77 (m, 1H), 8.34 (s, 1H), 8.02 (dd, J = 7.9, 4.6 Hz, 1H), 7.70 (s, 1H), 3.61 (s, 8H), 3.12–3.01 (m, 2H). LCMS (ESI) 428.00 [M+H] + . HPLC purity at 254 nm, 99.8%.

[0259] Example 66: 9-Morpholino-7-(trifluoromethyl)pyrido[2,3-b]phenazine-5,12-dione (66). General Protocol J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium (III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-morpholino-2-(trifluoromethyl)aniline (24.6 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-morpholino-7-(trifluoromethyl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (16.6 mg, 40% yield). 1H NMR (400MHz, methanol-d4) δ9.12(s,1H),8.84(d,J=7.9Hz,1H),8.31(d,J=2.4Hz,1H),8.00(dd,J=8.1,4.1Hz,1H),7.66(d,J=2. 8Hz, 1H), 4.01–3.91 (m, 2H), 3.88 (dt, J = 10.6, 4.7Hz, 2H), 3.74 (t, J = 5.7Hz, 1H), 3.66 (t, J = 4.9Hz, 2H), 3.45–3.38 (m, 1H). LCMS(ESI)415.05[M+H] + . HPLC purity at 254 nm, 100%.

[0260] Example 68: 9-(4-(tert-butyl)piperazin-1-yl)pyrido[2,3-b]phenazine-5,12-dione (68). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-(4-(tert-butyl)piperazin-1-yl)aniline (23.3 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-(4-(tert-butyl)piperazin-1-yl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (12 mg, 30% yield). 1 H NMR (400MHz, methanol-d4) δ9.14(dd,J=4.7,1.7Hz,1H),8.85(dd,J=8.0,1.7Hz,1H),8.24(d,J=9.5Hz,1H),8.10–7.9 7(m,2H),7.57(d,J=2.8Hz,1H),4.50(d,J=21.8Hz,2H),3.81(d,J=32.5Hz,2H),3.61–3.40(m,4H),1.55(s,9H). LCMS(ESI)402.20[M+H] + HPLC purity at 254 nm, 98.4%.

[0261] Example 69: 9-(4-(4-methoxyphenyl)piperazin-1-yl)pyrido[2,3-b]phenazine-5,12-dione (69). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-(4-(4-methoxyphenyl)piperazin-1-yl)aniline (28.3 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-(4-(4-methoxyphenyl)piperazin-1-yl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (22.0 mg, 49% yield). 1 H NMR (400MHz, chloroform-d) δ9.20(dd,J=4.6,1.7Hz,1H),8.81(dd,J=7.9,1.7Hz,1H),8.29(d,J=9.6Hz,1H),7.92–7.78(m,2H),7 .61(d,J=2.9Hz,1H),7.03–6.92(m,2H),6.92–6.82(m,2H),3.79(s,3H),3.78–3.73(m,4H),3.29(dd,J=6.4,3.8Hz,4H). LCMS(ESI)451.90[M+H] + HPLC purity at 254 nm, 99.3%.

[0262] Example 70: 9-((3aS,5S,7aS)-octahydro-7aH-2,5-inden-7a-yl)pyrido[2,3-b]phenazine-5,12-dione (70). General Protocol J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.), and 4-((3aS,5S,7aS)-octahydro-7aH-2,5-inden-7a-yl)aniline (22.7 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80 °C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL), and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred for 2 h at 90° C. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-((3aS,5S,7aS)-octahydro-7aH-2,5-oxodimethanoinden-7a-yl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (24.1 mg, 61% yield). 1 H NMR (400MHz, chloroform-d) δ9.26 (s, 1H), 8.86 (d, J = 7.9Hz, 1H), 8.53–8.37 (m, 2H), 8.20 (dd, J = 9.0, 2.2Hz, 1H) ,7.89(dd,J=7.8,4.3Hz,1H),2.21(s,3H),2.15(s,2H),2.07(d,J=2.9Hz,4H),1.85(q,J=12.6Hz,6H). LCMS(ESI)396.25[M+H] + HPLC purity at 254 nm, 95.8%.

[0263] Example 71: 9-(Thiazol-2-yl)pyrido[2,3-b]phenazine-5,12-dione (71). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-(thiazol-2-yl)aniline (17.6 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-(thiazol-2-yl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (22.7 mg, 66% yield). 1 H NMR (400MHz, chloroform-d) δ9.26 (dd, J=4.6, 1.8Hz, 1H), 9.01 (d, J=2.0Hz, 1H), 8.83 (ddd, J=15.7, 8.5, 1.9Hz ,2H),8.55(d,J=8.9Hz,1H),8.04(d,J=3.2Hz,1H),7.88(dd,J=8.0,4.5Hz,1H),7.56(d,J=3.2Hz,1H). LCMS(ESI)344.95[M+H] + . HPLC purity at 254 nm, 100%.

[0264] Example 72: 5,12-Dioxy-N,N-dipropyl-5,12-dihydropyrido[2,3-b]phenazine-9-sulfonamide (72). Follow General Scheme J. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium (III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-amino-N,N-dipropylbenzenesulfonamide (25.6 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 5,12-dioxy-N,N-dipropyl-5,12-dihydropyrido[2,3-b]phenazine-9-sulfonamide as a dark purple solid (21.6 mg, 51% yield). 1H NMR (400MHz, chloroform-d) δ9.30 (dd, J=4.6, 1.8Hz, 1H), 9.01 (dd, J=2.0, 0.6Hz, 1H), 8.88 (dd, J=8.0, 1.7Hz, 1H), 8.63 (dd, J=9.0, 0.6 Hz,1H),8.38(dd,J=8.9,2.0Hz,1H),7.92(dd,J=8.0,4.6Hz,1H),3.31–3.17(m,4H),1.70–1.53(m,4H),0.90(t,J=7.4Hz,6H). LCMS(ESI)424.95[M+H] + HPLC purity at 254 nm, 96.8%.

[0265] Example 73: 9-Cyclopropylpyrido[2,3-b]phenazine-5,12-dione (73). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-cyclopropylaniline (13.3 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and then the residue was purified by column chromatography to recover 9-cyclopropylpyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (21.0 mg, 70% yield). 1 H NMR (400MHz, chloroform-d) δ9.25(dd,J=4.6,1.8Hz,1H),8.85(dd,J=8.0,1.7Hz,1H),8.38(d,J=8.9Hz,1H),8.13(d,J=2.0Hz,1H), 7.87(dd,J=8.0,4.5Hz,1H),7.79(dd,J=8.9,2.0Hz,1H),2.22(tt,J=8.2,5.0Hz,1H),1.37–1.24(m,2H),1.12–0.94(m,2H). LCMS(ESI)302.00[M+H] + HPLC purity at 254 nm, 95.9%.

[0266] Example 74: 9-cyclohexylpyrido[2,3-b]phenazine-5,12-dione (74). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium (III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-cyclohexylaniline (17.5 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-cyclohexylpyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (23.3 mg, 68% yield). 1 H NMR (400MHz, chloroform-d) δ9.25 (dd, J=4.6, 1.8Hz, 1H), 8.85 (dd, J=7.9, 1.7Hz, 1H), 8. 41(d,J=8.8Hz,1H),8.34(dt,J=2.1,0.7Hz,1H),7.96(dd,J=9.0,1.9Hz,1H),7. 87(dd,J=7.9,4.5Hz,1H),2.96–2.75(m,1H),2.07(d,J=12.0Hz,2H),1.95(d,J= 12.5Hz, 2H), 1.84 (dd, J=12.9, 3.1Hz, 1H), 1.63–1.43 (m, 4H), 1.40–1.27 (m, 1H). LCMS (ESI) 344.05 [M+H] + . HPLC purity at 254 nm, 100%.

[0267] Example 75: 9-((2-(Diethylamino)ethyl)amino)pyrido[2,3-b]phenazine-5,12-dione (75). According to General Procedure J, 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and N 1A solution of -(2-(diethylamino)ethyl)benzene-1,4-diamine (20.7 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80 °C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90 °C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-((2-(diethylamino)ethyl)amino)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (7.1 mg, 19% yield). 1 H NMR (400MHz, methanol-d4) δ9.18–9.08(m,1H),8.84(dd,J=8.0,1.7Hz,1H),8.14(d,J=9.3Hz,1H),8.01(dd,J=8.0,4.7Hz,1H),7.61(dd ,J=9.3,2.6Hz,1H),7.24(d,J=2.6Hz,1H),3.86(t,J=6.6Hz,4H),3.55(t,J=6.6Hz,2H),3.45–3.37(m,4H),1.41(t,J=7.3Hz,6H). LCMS(ESI)376.10[M+H] + HPLC purity at 254 nm, 97.5%.

[0268] Example 76: 2-(4-Methylpiperazin-1-yl)benzo[b]phenazine-6,11-dione (76). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-(4-methylpiperazin-1-yl)aniline (19.1 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 2-(4-methylpiperazin-1-yl)benzo[b]phenazine-6,11-dione as a dark purple solid (26.9 mg, 75% yield). 1H NMR (400 MHz, methanol-d4) δ8.44 (ddd, J = 5.5, 4.2, 2.9 Hz, 2H), 8.21 (d, J = 9.5 Hz, 1H), 8.08-7.91 (m, 3H), 7.53 (d, J = 2.9 Hz, 1H), 4.65-4.21 (m, 2H), 3.92-3.37 (m, 4H), 2.68 (s, 3H). LCMS (ESI) 359.15 [M+H] + . HPLC purity at 254 nm, 98.8%.

[0269] Example 77: 2-(4-(tert-butyl)piperazin-1-yl)benzo[b]phenazine-6,11-dione (77). General Protocol J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-(4-(tert-butyl)piperazin-1-yl)aniline (23.3 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 2-(4-(tert-butyl)piperazin-1-yl)benzo[b]phenazine-6,11-dione as a dark purple solid (28.8 mg, 72% yield). 1 H NMR (400 MHz, chloroform-d) δ8.49 (t, J = 7.4 Hz, 2H), 8.36 (d, J = 9.5 Hz, 1H), 7.99-7.86 (m, 2H), 7.70 (d, J = 9.5 Hz, 1H), 7.63 (s, 1H), 4.15 (s, 2H), 3.83 (s, 4H), 3.05 (s, 2H), 1.52 (s, 9H). LCMS (ESI) 401.05 [M+H] + . HPLC purity at 254 nm, 99.6%.

[0270] Example 78: 9-(1-Methylpiperidin-4-yl)pyrido[2,3-b]phenazine-5,12-dione (78). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-(1-methylpiperidin-4-yl)aniline (19.1 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-(1-methylpiperidin-4-yl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (16.5 mg, 46% yield). 1 H NMR (400MHz, DMSO-d6) δ9.17(dd,J=4.6,1.7Hz,1H),8.70(dd,J=7.9,1.7Hz,1H),8.42(d,J=8.8Hz,1H),8.23(d,J=2.0Hz,1H),8.08(dd,J=8. 8,2.0Hz,1H),7.99(dd,J=7.9,4.6Hz,1H),3.63(d,J=12.0Hz,2H),3.58–3.39(m,3H),2.88(s,3H),2.26(d,J=13.8Hz,2H),2.18–2.03(m,2H). LCMS(ESI)359.15[M+H] + HPLC purity at 254 nm, 98.5%.

[0271] Example 79: 9-(4-Methyl-1,4-diazepan-1-yl)pyrido[2,3-b]phenazine-5,12-dione (79). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-(4-methyl-1,4-diazepan-1-yl)aniline (20.5 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-(4-methyl-1,4-diazepan-1-yl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (10.8 mg, 29% yield). 1 H NMR (400MHz, methanol-d4) δ9.13(dd,J=4.7,1.7Hz,1H),8.83(dd,J=8.0,1.7Hz,1H),8.14(d,J=9.6Hz,1H),8.01( dd,J=8.0,4.7Hz,1H),7.89(dd,J=9.6,2.9Hz,1H),7.34(d,J=2.8Hz,1H),4.31–3.41(m,10H),3.06(s,3H). LCMS(ESI)374.15[M+H] + HPLC purity at 254 nm, 97.4%.

[0272] Example 80: 9-(Pyrrolidin-1-yl)-10-(trifluoromethyl)pyrido[2,3-b]phenazine-5,12-dione (80). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-(pyrrolidin-1-yl)-3-(trifluoromethyl)aniline (23.0 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-(pyrrolidin-1-yl)-10-(trifluoromethyl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (7.2 mg, 18% yield). 1 H NMR (400 MHz, methanol-d4) δ 9.11 (d, J = 4.5 Hz, 1H), 8.81 (dd, J = 7.9, 1.7 Hz, 1H), 8.12 (d, J = 9.8 Hz, 1H), 8.03-7.91 (m, 2H), 3.76 (s, 4H), 2.14-2.03 (m, 4H). LCMS (ESI) 399.00 [M+H] + HPLC purity at 254 nm, 99.2%.

[0273] Example 81: 9-(Pyrrolidin-1-yl)-8-(trifluoromethyl)pyrido[2,3-b]phenazine-5,12-dione (81). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 4-(pyrrolidin-1-yl)-3-(trifluoromethyl)aniline (23.0 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-(pyrrolidin-1-yl)-8-(trifluoromethyl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (6.0 mg, 15% yield). 1H NMR (400MHz, methanol-d4) δ9.12 (dd, J=4.7, 1.7Hz, 1H), 8.84 (dd, J=8.0, 1.7Hz, 1H), 8.61 (s, 1H), 8.01 (dd, J = 8.0, 4.7Hz, 1H), 7.57 (s, 1H), 3.70 (t, J = 6.3Hz, 4H), 2.23–2.08 (m, 4H). LCMS(ESI)399.00[M+H] + HPLC purity at 254 nm, 98.4%.

[0274] Example 82: 10-Fluoro-9-(4-methylpiperidin-1-yl)pyrido[2,3-b]phenazine-5,12-dione (82). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 3-fluoro-4-(4-methylpiperidin-1-yl)aniline (20.8 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 10-fluoro-9-(4-methylpiperidin-1-yl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (13.1 mg, 35% yield). 1 H NMR (400MHz, methanol-d4) δ9.12(dd,J=4.6,1.7Hz,1H),8.85(dd,J=8.0,1.7Hz,1H),8.12–8.06(m,1H),8.04–7.93(m,2H),4.09 (d,J=12.8Hz,2H),3.24(t,J=12.6Hz,2H),1.88(d,J=13.1Hz,2H),1.75(s,1H),1.52–1.41(m,2H),1.06(d,J=6.5Hz,3H). LCMS(ESI)377.15[M+H] + HPLC purity at 254 nm, 98.5%.

[0275] Example 83: 8-Fluoro-9-(4-methylpiperidin-1-yl)pyrido[2,3-b]phenazine-5,12-dione (83). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 3-fluoro-4-(4-methylpiperidin-1-yl)aniline (20.8 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 8-fluoro-9-(4-methylpiperidin-1-yl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (0.98 mg, 26% yield). 1 H NMR (400MHz, methanol-d4) δ9.12(dd,J=4.7,1.7Hz,1H),8.82(dd,J=7.9,1.7Hz,1H),8.00(dd,J=7.9,4.6Hz,1H),7.88(s,1H),7.57(s,1H),3. 86(d,J=12.3Hz,2H),3.03–2.80(m,2H),1.86(d,J=12.9Hz,2H),1.75–1.61(m,1H),1.49(qd,J=12.3,3.8Hz,2H),1.07(d,J=6.5Hz,3H). LCMS(ESI)377.15[M+H] + . HPLC purity at 254 nm, 98.8%.

[0276] Example 84: 9-(4-Acetylpiperazin-1-yl)-10-(trifluoromethyl)pyrido[2,3-b]phenazine-5,12-dione (84). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 1-(4-(4-amino-2-(trifluoromethyl)phenyl)piperazin-1-yl)ethan-1-one (28.7 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-(4-acetylpiperazin-1-yl)-10-(trifluoromethyl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (22.3 mg, 49% yield). 1 H NMR (400MHz, chloroform-d) δ9.28 (dd, J=4.6, 1.8Hz, 1H), 8.85 (dd, J=8.0, 1.8Hz, 1H), 8.46 (d, J=9.6Hz, 1 H),7.93–7.82(m,2H),3.92(s,2H),3.79–3.73(m,2H),3.56(dd,J=11.6,6.9Hz,2H),2.23(s,3H). LCMS(ESI)456.10[M+H] + HPLC purity at 254 nm, 99.0%.

[0277] Example 85: 9-(4-Acetylpiperazin-1-yl)-8-(trifluoromethyl)pyrido[2,3-b]phenazine-5,12-dione (85). General Scheme J was followed. A solution of 6,7-dichloro-5,8-quinolinedione (0.1 eq.), cerium(III) chloride heptahydrate (CeCl3·7H2O, 0.11 eq.) and 1-(4-(4-amino-2-(trifluoromethyl)phenyl)piperazin-1-yl)ethan-1-one (28.7 mg, 0.1 mmol) in ethanol (2 mL) was stirred at 60-80°C for 2 h. Next, most of the ethanol was removed under vacuum. DMF (2 mL), H2O (10 μL) and sodium azide (13 mg, 0.2 mmol) were then added to the above reaction system. The mixture solution was stirred at 90°C for 2 h. The reaction mixture was cooled, the filtered precipitate was extracted with dichloromethane and concentrated, and the residue was then purified by column chromatography to recover 9-(4-acetylpiperazin-1-yl)-8-(trifluoromethyl)pyrido[2,3-b]phenazine-5,12-dione as a dark purple solid (21.0 mg, 46% yield). 1 HNMR (400MHz, methanol-d4) δ9.16 (dd, J=4.7, 1.7Hz, 1H), 8.89 (dd, J=8.0, 1.7 Hz, 1H), 8.78 (s, 1H), 8.29 (s, 1H), 8.04 (dd, J=8.0, 4.7 Hz,1H),3.86-3.79(m,4H),3.31-3.22(m,4H),2.22(s,3H). LCMS(ESI)456.10[M+H] + . HPLC purity at 254 nm, 99.8%.

[0278] Example III.

[0279] Representative compounds were tested in the National Cancer Institute's 60 cancer cell panel (NCI 60). Growth inhibition is reported in Tables 2, 3, 4, 5, and Figure 1 , Figure 2 , Figure 3 , Figure 4 , Figure 5 Shown in.

[0280] Table 2. Growth inhibition by Compound 1 in NCI60 cell line (% control).

[0281]

[0282]

[0283]

[0284] Table 3. Growth inhibition of compound 13 in NCI60 cell line (% control).

[0285]

[0286]

[0287]

[0288] Table 4. Growth inhibition of compound 49 in NCI60 cell line (% control).

[0289]

[0290]

[0291]

[0292] Table 5. Growth inhibition of Compound 50 in NCI60 cell line (% control).

[0293]

[0294]

[0295]

[0296] The present invention has now been fully described, and it will be appreciated by those skilled in the art that the present invention can be carried out in a wide and equivalent range of conditions, formulations, and other parameters without affecting the scope of the present invention or any embodiment thereof. All patents, patent applications, and publications cited herein are fully incorporated herein by reference in their entirety.

[0297] Incorporation by reference

[0298] The entire disclosure of each patent document and scientific article mentioned herein is incorporated by reference for all purposes.

[0299] Equivalent

[0300] Without departing from the spirit or essential features of the present invention, the present invention may be embodied in other specific forms. Therefore, the foregoing embodiments are considered to be illustrative in all respects, rather than limiting the invention described herein. Therefore, the scope of the present invention is indicated by the appended claims rather than by the foregoing description, and all changes falling within the equivalent meaning and scope of the claims are intended to be included therein.

Claims

1. A compound contained in formula VII, IX, XI or XIII: Including pharmaceutically acceptable salts, solvates and / or prodrugs thereof; wherein: R 1 is selected from the group consisting of: hydrogen or optionally substituted alkyl or halogen; R 2 Selected from the group consisting of alkyl, optionally substituted C4-C8 cycloalkyl, optionally substituted C4-C8 heteroalkyl, optionally substituted C4-C8 heterocycloalkyl, optionally substituted C5-C6 aryl, and optionally substituted C4-C6 heteroaryl; R 3a , R 3b , R 3c or R 3d selected from the group consisting of hydrogen, halogen, methyl, methoxy, trifluoromethyl, hydroxyl or cyano; R 4 selected from the group consisting of alkyl or heteroalkyl; A includes cycloalkyl, heterocycloalkyl, aryl or heteroaryl; X is selected from the group consisting of hydrogen, halogen, amino, heterocycloalkyl or hydroxy; Y is an O atom, a S atom or a N atom; m is an integer selected from 0-9.

2. The compound according to claim 1, wherein R 1 Selected from the group consisting of hydrogen, halogen, alkyl, heteroalkyl, optionally substituted C6-C 14 aryl and optionally substituted 5- to 14-membered heteroaryl.

3. The compound according to claim 1, wherein R 2 is optionally substituted and selected from the group consisting of:

4. The compound according to claim 1, wherein R 3a , R 3b , R 3c or R 3d is optionally mono- or poly-substituted and selected from the group consisting of:

5. The compound according to claim 1, wherein R 4 is optionally substituted and selected from the group consisting of:

6. The compound according to claim 1, wherein X is optionally substituted and is selected from the group consisting of:

7. A pharmaceutical composition comprising the compound according to any one of claims 1 to 5.

8. Use of the pharmaceutical composition according to claim 7 in the preparation of a medicament for treating, ameliorating or preventing leukemia, non-small cell lung cancer, colon cancer, CNS cancer, melanoma, ovarian cancer, gastric cancer, prostate cancer or breast cancer in a patient.

9. The use according to claim 8, wherein The patient is a human patient.

10. The use according to claim 8, further comprising administering one or more anti-cancer agents to the patient.

11. The use according to claim 10, wherein The anti-cancer agent is a chemotherapeutic agent.

12. The use according to claim 10, wherein The anti-cancer agent is radiation therapy.

13. A kit comprising a compound according to any one of claims 1-6 and instructions for administering the compound to a patient having leukemia, non-small cell lung cancer, colon cancer, CNS cancer, melanoma, ovarian cancer, gastric cancer, prostate cancer or breast cancer.

14. The kit of claim 13, further comprising one or more anti-cancer agents.

15. The kit according to claim 14, wherein The compounds are administered together with one or more anti-cancer agents.

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