Furanoquinolinediones as inhibitors of TDP2

Abstract

The present invention relates to furanoquinolinediones as inhibitors of TDP2. The present invention discloses compounds of formula I and pharmaceutically acceptable salts thereof. The variable X is disclosed herein. 1 , X 2 and R 1‑4 The compounds are useful for treating cancer and related proliferative diseases. Pharmaceutical compositions containing the compounds of formula I and methods of treatment comprising administering the compounds of formula I are also disclosed.

Description

[0001] This application is a divisional application of Chinese Patent Application No. 201680014742.7, filed on January 8, 2016, entitled “Furanoquinolinediones as inhibitors of TDP2”.

[0002] Government Support Statement

[0003] This invention was made in part with government support from the National Institutes of Health. The government has certain rights in this invention.

[0004] Citation of Related Applications

[0005] This application claims priority to U.S. Provisional Application No. 62 / 100,968, filed January 8, 2015, which is incorporated herein by reference in its entirety. Technical Field

[0006] The present application relates to furanoquinolinedione and pharmaceutically acceptable salts thereof as inhibitors of TDP2 and also to the use of the furanoquinolinedione for treating cancer and related proliferative diseases. Background Art

[0007] Topoisomerases are a family of enzymes necessary for relaxing supercoiled DNA, which is required for transcription, replication, and chromosome segregation. Topoisomerases cut supercoiled nucleic acids, which relieve torsional strain and then rejoin the cut strands. The important role of topoisomerases in DNA replication makes these enzymes a major target for cancer therapy. Topoisomerase inhibitors, such as camptothecin and topotecan for topoisomerase 1 (Top1), and etoposide and doxorubicin for topoisomerase 2 (Top2), trap the topoisomerase-DNA cleavage complex, thereby preventing replication and initiating destruction by apoptosis.

[0008] Tyrosyl-DNA phosphodiesterase (TDP) plays an important role in the repair of trapped topoisomerase-DNA cleavage complexes. TDP exists in two forms: TDP1, which is used to disassemble trapped Top1-DNA cleavage complexes, and TDP2, which is used to disassemble Top2-DNA cleavage complexes.

[0009] Since TDP can repair a wide spectrum of nucleic acid damage, inhibition of TDP is an attractive target for therapeutic treatment of cancer, possibly as a sole therapeutic agent. It has also been noted that TDP2 deficiency can enhance the antiproliferative activity of Top2 inhibitors in cells with defective cellular checkpoints, and therefore, TDP2 inhibitors can provide a synergistic effect as a combination therapy with inhibitors of Top2 (such as etoposide and doxorubicin). It can be expected that the use of inhibitors of Top1 and inhibitors of TDP1 has a similar synergistic effect. Therefore, identifying effective inhibitors of TDP1 and / or TDP2 will be important in providing new methods for treating cancer. Summary of the invention

[0010] Described herein are inhibitors of TDP2, methods of making them, compositions containing the compounds, and methods of using the compounds. In a first aspect, provided are compounds of Formula I and pharmaceutically acceptable salts of the compounds of Formula I,

[0011]

[0012] In Formula I, the following conditions are met.

[0013] X 1 Is N or CR 5 .

[0014] X 2 Is N or CR 6 .

[0015] X 1 or X 2 At least one of them is N.

[0016] R 1 It is hydroxyl, C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, C 2 -C 6 Alkenyloxy, C 2 -C 6 Alkynyloxy, or C 3 -C 7 Cycloalkoxy, where R 1 One or more methylene units in the alkyl, alkenyl, or alkynyl portion of the present invention are optionally or independently replaced by -O-, -S-, or -N(R 7 )-replaced, and R other than hydroxyl 1 At each occurrence, 0-3 independently selected from halogen, hydroxy, cyano, =N, =NOR 7 , -CO 2H, -(CO)-OC 1 -C 6 Alkyl, -C(O)NR 7 R 8 and-WP(O)YR 9 ZR 10 substituted by a substituent.

[0017] or R 1 It may be -OAB, wherein O is an oxygen atom.

[0018] A is a linker consisting of a bond, an alkylene chain of 1 to 6 carbons, or a phenylene group.

[0019] B is phenyl, or a 5- or 6-membered heterocyclic ring having 1, 2 or 3 ring atoms independently selected from N, O and S, wherein B is independently selected from halogen, hydroxy, cyano, amino, -SH, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, C 1 -C 6 Alkoxy, C 2 -C 6 Alkanoyl, C 1 -C 6 Thioalkyl, C 1 -C 6 Haloalkyl, C 1 -C 6 Haloalkoxy, -(C 0 -C 6 alkyl)cycloalkyl, -(C 0 -C 6 alkyl)CO 2 H, -(C 0 -C 6 (alkyl)-(CO)-OC 1 -C 6 Alkyl, -(C 0 -C 6 Alkyl)C(O)NR 7 R 8 、-(C 0 -C 6 Alkyl)NR 7 C(O)R 8 、-(C 1 -C 6 Alkyl) alkoxy, -(C 1 -C 6 Alkyl)OH, -(C 0 -C 6 Alkyl)NR7 R 8 、-SO 2 -C 1 -C 6 Alkyl and -(C 0 -C 6 Alkyl)-WP(O)YR 9 ZR 10 Substitute with 0-3 substituents of your choice.

[0020] Y 1 is O, NH or S.

[0021] Y 2 Is N or CR 2 .

[0022] R 2 is hydrogen, halogen, C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Haloalkyl, C 1 -C 6 Haloalkoxy, -(C 0 -C 6 alkyl)cycloalkyl, -(C 0 -C 6 Alkyl)NR 7 R 8 or phenyl, each R except hydrogen and hydroxy 2 Each occurrence is independently selected from halogen, hydroxyl, C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Haloalkyl and C 1 -C 6 The haloalkoxy group is substituted with 0 to 3 groups selected from the group consisting of:

[0023] Or, R 2 is a group -JQ, where J is an alkylene linker of 1 to 4 carbons, where any -CH 2 -groups may be optionally replaced by -C(O)O-, -C(O)NH-, -C(O)NR 11 or -C(O)- substitution.

[0024] Q is for C 1 -C 6 Alkyl, C 1 -C 6alkylamino, aryl or heteroaryl, each of which is unsubstituted or substituted independently from halogen, hydroxy, amino, cyano, C 1 -C 4 Alkyl, C 1 -C 4 Alkoxy, C 1 -C 2 Haloalkyl and C 1 -C 2 The haloalkoxy group is substituted with one or more groups selected from the group consisting of haloalkoxy and alkyl alkyl.

[0025] W, Y and Z are independently a bond or O at each occurrence.

[0026] R 3 , R 4 , R 5 and R 6 is independently selected at each occurrence from hydrogen, halogen, cyano, amino, C 1 -C 6 Alkyl, -(C 0 -C 6 alkyl)cycloalkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Haloalkyl and C 1 -C 6 Halogenated alkoxy.

[0027] R 7 , R 8 , R 9 and R 10 is independently selected at each occurrence from hydrogen, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, -(C 0 -C 6 alkyl)cycloalkyl and C 1 -C 6 haloalkyl, and any R bonded to the same nitrogen atom 7 and R 8 can be combined to form a 4- to 7-membered heterocycloalkyl group, which is supported by 0 to 2 groups selected from hydroxy, halogen, C 1 -C 4 Alkyl, C 1 -C 4 Alkoxy and C 2 -C 4 The alkanoyl group is substituted with a substituent of choice.

[0028] R 11 It is C 1 -C 6Alkyl or C 1 -C 6 Alkylamino.

[0029] Where the compound is not:

[0030]

[0031] Also disclosed is a pharmaceutical composition comprising a compound or salt of Formula I and a pharmaceutically acceptable carrier.

[0032] Also disclosed is a method of treating a disorder responsive to TDP2 inhibition comprising the step of administering a compound or a salt thereof to a patient in need thereof.

[0033] Also disclosed are methods of treating cancer (including glioma (glioblastoma, malignant glioma), acute myelogenous leukemia, acute myeloid leukemia, myelodysplastic / myeloproliferative tumors, sarcomas, chronic myelomonocytic leukemia, non-Hodgkin's lymphoma, astrocytomas, melanomas, non-small cell lung cancer, small cell lung cancer, cervical cancer, rectal cancer, ovarian cancer, cholangiocarcinomas, chondrosarcomas, or colon cancer), comprising administering to a patient in need of such treatment a therapeutically effective amount of a compound or salt of Formula I. DETAILED DESCRIPTION

[0034] the term

[0035] The compounds are described using standard terminology. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0036] The terms "a" and "an" do not denote a limitation of quantity, but rather denote the presence of at least one of the referenced item. The term "or" means "and / or." The terms "comprising," "having," "including," and "containing" are to be construed as open-ended terms (i.e., meaning "including, but not limited to").

[0037] Recitation of ranges of values ​​is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. All endpoints of ranges are included within the range and are independently combinable.

[0038] All methods described herein can be performed in an appropriate order, unless otherwise specified herein or otherwise explicitly contradicted by context. The use of any and all examples or exemplary language (e.g., "such as") is intended only to better illustrate the present invention and is not intended to limit the scope of the present invention unless otherwise specified. Any language in the specification should not be construed as indicating that any non-claimed element is essential to the practice of the present invention used herein. Unless otherwise defined, the technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art of the present disclosure.

[0039] In addition, the present disclosure covers all variations, combinations and permutations in which one or more limitations, elements, clauses and descriptive terms from one or more of the listed claims are introduced into another claim. For example, any claim that is dependent on another claim may be amended to include one or more limitations found in any other claim that is dependent on the same base claim. When elements are presented in list form, such as in Markush group form, each subgroup of the elements is also disclosed, and any element may be removed from the group.

[0040] All compounds are understood to include all possible isotopes of atoms occurring in the compound. Isotopes include those atoms having the same atomic number but different mass numbers. As a general example and without limitation, isotopes of hydrogen include tritium and deuterium, and isotopes of carbon include 11 C. 13 C and 14 C.

[0041] Formula I includes all pharmaceutically acceptable salts of Formula I.

[0042] The open-ended term "comprising" includes intermediates and the closed terms "consisting essentially of" and "consisting of.

[0043] The term "substituted" means that any one or more hydrogens on the specified atom or group are replaced by a group selected from the specified group, provided that the normal valence of the specified atom is not exceeded. When the substituent is an oxy group (i.e., =O), then 2 hydrogens on the atom are replaced. When the aromatic moiety is substituted by an oxy group, the aromatic ring is replaced by a corresponding partially unsaturated ring. For example, a pyridyl group substituted by an oxy group is a pyridone. These combinations are allowed only when the combination of substituents and / or variables produces a stable compound or a useful synthetic intermediate. A stable compound or a stable structure means that the compound is strong enough to be isolated from a reaction mixture and subsequently formulated into an effective therapeutic agent.

[0044] Suitable groups that may be present at an "optionally substituted" position include, but are not limited to, for example, halogen, cyano, hydroxy, amino, nitro, oxo, azido, alkanoyl (such as C 2 -C 6 Alkanoyl, such as acyl, etc. (-(CO)alkyl)); carboxamido; alkylcarboxamide; alkyl, alkoxy, alkylthio (including those with one or more thioether bonds), alkylsulfinyl (including those with one or more sulfinyl bonds), alkylsulfonyl (including those with one or more sulfonyl bonds), mono- and di-aminoalkyl (including groups with one or more N atoms), one or more methylene groups in all of the above optional alkyl substituents may be replaced by oxygen or -NH- and have about 1 to about 8, about 1 to about 6 or 1 to about 4 carbon atoms, cycloalkyl; phenyl; phenylalkyl, benzyl is an exemplary phenylalkyl, phenylalkoxy, benzyloxy is an exemplary phenylalkoxy. Alkylthio and alkoxy are attached to the position they replace through sulfur or oxygen atoms, respectively.

[0045] A dash ("-") that is not between two letters or symbols is used to indicate the point of attachment of a substituent.

[0046] "Alkyl" includes both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms, typically 1 to about 8 carbon atoms. As used herein, the term C 1 -C 6 Alkyl refers to an alkyl group having 1, 2, 3, 4, 5 or 6 carbon atoms. Other embodiments include alkyl groups having 1 to 8 carbon atoms, 1 to 4 carbon atoms or 1 or 2 carbon atoms, such as C 1 -C 8 Alkyl, C 1 -C 4 Alkyl and C 1 -C 2 Alkyl. In this article, C 0 -C n When alkyl is used together with other groups, for example the indicated group -C 0 -C2 Alkyl (phenyl), in this case, the phenyl group is directly attached via a single covalent bond (C 0 alkyl) or through an alkyl chain having the specified number of carbon atoms, in this case 1, 2, 3 or 4 carbon atoms. The alkyl group may also be attached through other groups such as heteroatoms, as in -OC 0 -C 4 Alkyl (C 3 -C 7 Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, 3-methylbutyl, tert-butyl, n-pentyl, and sec-pentyl.

[0047] "Alkenyl" is a branched or straight chain aliphatic hydrocarbon group having one or more carbon-carbon double bonds that may occur at any stable point along the chain having the specified number of carbon atoms. Examples of alkenyl groups include, but are not limited to, ethenyl and propenyl.

[0048] "Alkynyl" refers to a branched or straight chain aliphatic hydrocarbon group having one or more dicarbon-carbon triplet bonds which may occur at any stable point along the chain having the specified number of carbon atoms.

[0049] "Alkoxy" refers to an alkyl group with a specified number of carbon atoms as defined above that is covalently bonded to its substituted group through an oxygen bridge (-O-). The example of alkoxy includes but is not limited to methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, 2-butoxy, tert-butoxy, n-pentyloxy, 2-pentyloxy, 3-pentyloxy, isopentyloxy, neopentyloxy, n-hexyloxy, 2-hexyloxy, 3-hexyloxy and 3-methylpentyloxy. Similarly, "alkylthio" or "thioalkyl" refers to an alkyl group with a specified number of carbon atoms as defined above that is covalently bonded to its substituted group through a sulphur bridge (-S-). Similarly, "alkenyloxy", "alkynyloxy" and "cycloalkoxy" refer to alkenyl, alkynyl and cycloalkyl groups that are covalently bonded to their substituted groups through an oxygen bridge (-O-) in each case.

[0050] "Alkanoyl" refers to an alkyl group as defined above having the specified number of carbon atoms covalently bonded to its substituent through a keto group (-C(O)-). The carbon of the keto group is included in the number of carbon atoms in the alkanoyl group, i.e., C 2 Alkanoyl is -C(O))CH 3 .

[0051] An "alkylene" group is a chain of one or more methylene groups with a point of attachment at each end such that it can link two other groups, i.e., an alkylene group is -(CH 2 ) n -.

[0052] "Cycloalkyl" is a saturated hydrocarbon ring radical having a specified number of carbon atoms, typically 3 to about 7 carbon atoms. Examples of cycloalkyl radicals include cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl and bridged or caged saturated ring radicals such as norbornane or adamantane. "-(C 0 -C n Alkyl)cycloalkyl" is a single covalent bond (C 0 ) or a cycloalkyl group attached to the position where it is substituted through an alkylene linker having 1 to n carbon atoms.

[0053] "Halo" or "halogen" refers to fluoro, chloro, bromo or iodo.

[0054] "Heteroaryl" is a stable monocyclic aromatic ring having a specified number of ring atoms, containing 1 to 3 or, in some embodiments, 1 to 2 heteroatoms selected from N, O and S, the remaining ring atoms being carbon; or a stable bicyclic or tricyclic ring system containing at least one 5- to 7-membered aromatic ring containing 1 to 3 or, in some embodiments, 1 to 2 heteroatoms selected from N, O and S, the remaining ring atoms being carbon. Monocyclic heteroaryl groups typically have 5 to 7 ring atoms. In some embodiments, bicyclic heteroaryl groups are 9 to 10-membered heteroaryl groups, i.e., groups containing 9 or 10 ring atoms, in which one 5- to 7-membered aromatic ring is fused to a second aromatic or non-aromatic ring. When the total number of S and O atoms in the heteroaryl group exceeds 1, these heteroatoms are not adjacent to each other. Preferably, the total number of S and O atoms in the heteroaryl group is not greater than 2. It is particularly preferred that the total number of S and O atoms in the aromatic heterocycle is not greater than 1. Heteroaryl groups include, but are not limited to, oxazolyl, piperazinyl, pyranyl, pyrazolopyrimidinyl, pyrazolyl, pyridizinyl, pyridinyl, pyrimidinyl, pyrrolyl, quinolinyl, tetrazolyl, thiazolyl, thienylpyrazolyl, thienyl, triazolyl, benzo[d]oxazolyl, benzofuranyl, benzothiazolyl, benzothienyl, benzoxadiazolyl, dihydrobenzodioxynyl, furanyl, imidazolyl, indolyl, and isoxazolyl.

[0055] "Heterocycle" is a saturated, unsaturated or aromatic cyclic group having the specified number of ring atoms, containing 1 to about 3 heteroatoms selected from N, O and S, the remainder of the ring atoms being carbon. Examples of heterocyclic groups include piperazinyl and thiazolyl.

[0056] "Heterocycloalkyl" is a saturated cyclic group having the specified number of ring atoms, containing 1 to about 3 heteroatoms selected from N, O and S, with the remainder of the ring atoms being carbon. Examples of heterocycloalkyl include tetrahydrofuranyl and pyrrolidinyl.

[0057] "Haloalkyl" refers to branched and straight chain alkyl groups having a specified number of carbon atoms, which are substituted with one or more halogen atoms, usually up to the maximum permitted number of halogen atoms. Examples of haloalkyl groups include, but are not limited to, trifluoromethyl, difluoromethyl, 2-fluoroethyl, and pentafluoroethyl.

[0058] "Haloalkoxy" is a haloalkyl group as defined above attached through an oxygen bridge (the oxygen of the alcohol group).

[0059] "Phenylene" is a benzene ring with two distinct points of attachment so that two other groups can be attached, i.e., phenylene is -(C 6 H 4 The points of attachment can be ortho, meta or para to each other.

[0060] "Pharmaceutical composition" refers to a composition comprising at least one active agent (eg, a compound or salt of Formula I) and at least one other substance (eg, a carrier). The pharmaceutical composition meets USFDA GMP (Good Manufacturing Practice) standards for human or non-human drugs.

[0061] "Carrier" refers to a diluent, excipient or vehicle for administering an active compound. "Pharmaceutically acceptable carrier" refers to a substance such as an excipient, diluent or vehicle that can be used to prepare a pharmaceutical composition that is generally safe, non-toxic and neither biologically nor otherwise undesirable, and includes carriers that are acceptable for veterinary use as well as human pharmaceutical use. "Pharmaceutically acceptable carrier" includes one and more than one such carrier.

[0062] "Patient" refers to a human or non-human animal in need of medical treatment. Medical treatment can include treatment of an existing condition, such as a disease or disorder, or diagnostic treatment. In some embodiments, the patient is a human patient.

[0063] "Provide" means to give, administer, sell, distribute, transfer (for profit or not), manufacture, mix or assign.

[0064] "Treatment" or "treating" refers to providing an active compound to a patient in an amount sufficient to measurably reduce any symptoms of cancer, slow cancer progression, or cause cancer regression. In certain embodiments, cancer treatment can be initiated before a patient develops symptoms of the disease.

[0065] A "therapeutically effective amount" of a pharmaceutical composition refers to an amount that, when administered to a patient, is effective to provide a therapeutic benefit, such as improving symptoms, reducing cancer progression, or causing cancer regression.

[0066] A significant change is any detectable change that is statistically significant in a standard parametric test of statistical significance, such as the Student's T test, with p < 0.05.

[0067] Chemical description

[0068] The compound of formula I may contain one or more asymmetric elements, such as stereocenters, stereoaxes, etc., such as asymmetric carbon atoms, so that the compound can exist in different stereoisomeric forms. These compounds may be, for example, racemates or optically active forms. For compounds with two or more asymmetric elements, these compounds may additionally be mixtures of diastereomers. For compounds with asymmetric centers, all optical isomers and mixtures thereof in pure form are encompassed. In these cases, a single enantiomer (i.e., optically active form) may be synthesized by asymmetric synthesis, from optically pure precursors, or by splitting the racemate to obtain. The splitting of the racemate may also be accomplished, for example, by conventional methods (such as crystallization in the presence of a splitting agent) or by using, for example, the chromatography of a chiral HPLC column. Regardless of the method for obtaining them, all forms are contemplated herein.

[0069] All forms of the active agent (eg, solvates, optical isomers, enantiomeric forms, polymorphs, free compounds and salts) may be employed alone or in combination.

[0070] The term "chiral" refers to a molecule having the property of non-superimposability of a mirror image partner.

[0071] "Stereoisomers" are compounds that have identical chemical constitution, but differ with regard to the arrangement of the atoms or groups in space.

[0072] "Diastereomers" are stereoisomers with two or more chiral centers, and their molecules are not mirror images of each other. Diastereomers have different physical properties, such as melting points, boiling points, spectral properties and reactivity. Mixtures of diastereomers can be separated under high-resolution analytical procedures (e.g., electrophoresis, crystallization in the presence of a resolving agent, or chromatography using, for example, a chiral HPLC column).

[0073] "Enantiomers" refers to two stereoisomers of a compound that are non-superimposable mirror images of each other. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, which may occur in the absence of stereoselectivity or stereospecificity in a chemical reaction or process.

[0074] Stereochemical definitions and conventions used herein generally follow SP Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., Stereochemistry of Organic Compounds (1994) John Wiley & Sons, Inc., New York. Many organic compounds exist in optically active forms, i.e., they have the ability to rotate the plane of plane-polarized light. In describing optically active compounds, the prefixes D and L or R and S are used to denote the absolute configuration of the molecule about its chiral center. The prefixes d and l or (+) and (-) are used to indicate the sign of the rotation of plane-polarized light by the compound, (-) and l meaning that the compound is levorotatory. Compounds with a (+) or d prefix are dextrorotatory.

[0075] A "racemic mixture" or "racemate" is an equimolar (or 50:50) mixture of two enantiomeric species that is optically devoid of activity. A racemic mixture may occur when there is no stereoselectivity or stereospecificity in a chemical reaction or process.

[0076] "Pharmaceutically acceptable salts" include derivatives of disclosed compounds, wherein the parent compound is modified by preparing inorganic and organic, non-toxic acid or base addition salts of the parent compound. Salts of the compounds of the present invention can be synthesized by a parent compound containing an alkaline or acidic part by conventional chemical methods. Typically, these salts can be prepared by reacting these compounds in free acid form with a stoichiometric amount of a suitable base (e.g., Na, Ca, Mg or K hydroxide, carbonate, bicarbonate, etc.), or by reacting these compounds in free base form with a stoichiometric amount of a suitable acid. This reaction is typically carried out in water or in an organic solvent or in a mixture of the two. Typically, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile are used where feasible. Salts of the compounds of the present invention also include solvates of the compounds and solvates of the salts of the compounds.

[0077] Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like. Pharmaceutically acceptable salts include, for example, conventional non-toxic salts and quaternary ammonium salts of the parent compound formed from non-toxic inorganic or organic acids. For example, conventional non-toxic acid salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, aminosulfonic acid, phosphoric acid, nitric acid, and the like; and salts derived from organic acids such as acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, methanesulfonic acid, esylic acid, besylic acid, p-aminobenzenesulfonic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, isethionic acid, HOOC-(CH 2 ) n -COOH (wherein n is 0-4) and other organic acids. Additional suitable salts can be listed, for example, in G. Steffen Paulekuhn, et al., Journal of Medicinal Chemistry 2007, 50, 6665 and Handbook of Pharmaceutically Acceptable Salts: Properties, Selection and Use, P. Heinrich Stahl and Camille G. Wermuth, Editors, Wiley-VCH, 2002.

[0078] TDP2 inhibitors

[0079] Disclosed herein are molecules that inhibit TDP2.

[0080] In addition to the compounds of Formula I shown in the Summary of the Invention section, the present disclosure also includes compounds wherein the variables such as A, B, X 1 , X 2 , W, Y, Z, R 1 To R 10 Compounds carrying the following definitions. The present disclosure includes all combinations of these definitions as long as stable compound results can be obtained. The present disclosure includes the following specific embodiments of formula (I).

[0081]

[0082] The present disclosure includes compounds of Formulae IA and IB and salts thereof.

[0083]

[0084] R 1 It is hydroxyl, C 1 -C 8 Alkoxy or –O-(C 0 -C 6 alkyl)cycloalkyl, wherein R 1 One or more methylene units in the alkoxy or alkyl portion of the 7 )-substitution, and R 1 At each occurrence, 0-3 independently selected from hydroxy, halogen, cyano, -CO 2 H, -(CO)-OC 1 -C 6 Alkyl and -WP(O)YR 9 ZR 10 or R 1 Yes - OAB.

[0085] B is phenyl or a 5- or 6-membered heteroaryl group having 1, 2 or 3 nitrogen ring atoms, wherein B is optionally substituted by 0-3 groups independently selected from halogen, hydroxy, cyano, amino, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, C 2 -C 6 Alkanoyl, C 1 -C 6 Alkoxy, C 1 -C 6 Haloalkyl, C 1 -C 6 Haloalkoxy, -(C 0 -C 6 alkyl)cycloalkyl, -(C 0 -C 6 alkyl)CO 2 H,-(C 0 -C 6 (alkyl)-(CO)-OC 1 -C 6 Alkyl, -(C 1 -C 6 Alkyl) alkoxy, -(C 1 -C 6 Alkyl)OH,-SO 2 -C 1 -C 6 Alkyl and -(C 0 -C 6 Alkyl)-WP(O)YR 9 ZR10 The selected substituent is substituted.

[0086] R 2 Halogen, C 1 -C 6 Alkyl, C 1 -C 6 haloalkyl, or -(C 0 -C 6 alkyl)cycloalkyl, or phenyl, wherein each occurrence of phenyl is replaced by 0 to 3 independent residues selected from halogen, hydroxy, C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Haloalkyl and C 1 -C 6 The haloalkoxy group is substituted with a selected group.

[0087] (B)R 3 and R 4 All are hydrogen.

[0088] (C)R 2 It is C 1 -C 6 Alkyl, C 1 -C 6 Haloalkyl, -(C 0 -C 6 alkyl)cycloalkyl, or phenyl, wherein each occurrence of phenyl is replaced by 0 to 3 independently selected from halogen, hydroxy, C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Haloalkyl and C 1 -C 6 The group selected from the group consisting of haloalkoxy and (D) R 1 By at least one -WP(O)YR 9 ZR 10 Substituents are substituted; and

[0089] W is a bond, and Y and Z are both O.

[0090] (E)R 2 It is methyl or phenyl.

[0091] (F)X 1 is N and X 2 It is CH.

[0092] (G)X 1 is CH and X 2It is N.

[0093] (H)R 1 It is C 1 -C 8 Alkoxy.

[0094] R 1 is phenoxy or pyridyloxy, each of which is optionally substituted by 0-3 independently selected from halogen, hydroxy, cyano, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, C 1 -C 6 Alkoxy, C 1 -C 6 Haloalkyl, C 1 -C 6 Haloalkoxy, -(C 0 -C 6 alkyl)cycloalkyl, -(C 0 -C 6 alkyl)CO 2 H, -(C 0 -C 6 (alkyl)-(CO)-OC 1 -C 6 Alkyl, -(C 1 -C 6 Alkyl) alkoxy, -(C 1 -C 6 Alkyl)OH, -SO 2 -C 1 -C 6 Alkyl and -(C 0 -C 6 Alkyl)-WP(O)YR 9 ZR 10 The selected substituent is substituted.

[0095] (I)R 1 is an alkoxy group, where R 1 One or more methylene units in the alkyl portion of the 7 )-substitution, and R 1 1 to 3 independently selected from hydroxyl and -WP(O)YR 9 ZR 10 wherein W is a bond; and Y and Z are both O.

[0096] (J)R 1 Yes - OAB.

[0097] (K)R 1 Yes - OAB;

[0098] A is a bond or an alkylene chain of 1 to 3 carbon atoms; and

[0099] B is phenyl or pyridyl, which is optionally substituted by 0-3 groups independently selected from halogen, hydroxy, cyano, C 1 -C 4 Alkyl, C 1 -C 4 Alkoxy, C 2 -C 4 Alkanoyl, C 1 -C 2 Haloalkyl, C 1 -C 2 Haloalkoxy, -(C 0 -C 2 (alkyl)-(CO)-OC 1 -C 6 Alkyl, -(C 0 -C 2 Alkyl)C(O)NR 7 R 8 、-(C 0 -C 2 Alkyl)NR 7 C(O)R 8 、-(C 1 -C 6 Alkyl)OH and -SO 2 -C 1 -C 2 The alkyl group is substituted with a selected substituent.

[0100] (L)R 1 Yes - OAB;

[0101] A is a bond; and

[0102] B is phenyl which is substituted with a substituent selected from hydroxy, halogen and cyano.

[0103] (M)R 1 is -OAB; and

[0104] B is triazolyl, pyrazolyl, imidazolyl, thienyl, dioxolyl, morpholinyl, piperazinyl or piperidinyl; each of said B is substituted by 0-3 independently selected from halogen, hydroxyl, cyano, amino, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, C 1-C 6 Alkoxy, C 2 -C 6 Alkanoyl, C 1 -C 2 Haloalkyl, C 1 -C 2 Haloalkoxy, -(C 0 -C 6 alkyl)CO 2 H, -(C 0 -C 6 (alkyl)-(CO)-OC 1 -C 6 Alkyl, -(C 0 -C 6 Alkyl)C(O)NR 7 R 8 、-(C 0 -C 6 Alkyl)NR 7 C(O)R 8 、-(C 1 -C 6 Alkyl)OH, -(C 0 -C 6 Alkyl)NR 7 R 8 and-SO 2 -C 1 -C 6 The alkyl group is substituted by a substituent selected from the group consisting of 7 and R 8 is hydrogen or C 1 -C 4 alkyl.

[0105] (N)R 2 is hydrogen, halogen, C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Haloalkyl, C 1 -C 6 Haloalkoxy, -(C 0 -C 6 alkyl)cycloalkyl, -(C 0 -C 6 Alkyl)NR 7 R 8 or phenyl, each R except hydrogen and halogen 2 At each occurrence, 0 to 3 independently selected from halogen, hydroxyl, C 1 -C 6 Alkyl, C 1 -C 6Alkoxy, C 1 -C 6 Haloalkyl and C 1 -C 6 The haloalkoxy group is substituted with a selected group.

[0106] (O)R 2 yes

[0107]

[0108] Where n is 1, 2, 3 or 4;

[0109] J 1 O, NH, NR 11 ;

[0110] Q 1 It is C 1 -C 6 Alkyl or C 1 -C 6 Alkylamino-;

[0111] R 12 There is no or one or more substituents independently selected from hydroxy, halogen, amino or cyano; and

[0112] R 13 Absent or 1 or 2 independently selected from C 1 -C 4 Alkyl and mono- or di-C 1 -C 6 Substituents of the alkylamino group.

[0113] The present disclosure includes one embodiment which is a pharmaceutical composition comprising a compound or salt of Formula I and a pharmaceutically acceptable carrier.

[0114] Pharmaceutical preparations

[0115] The compounds disclosed herein can be administered as pure chemicals, but are preferably administered as pharmaceutical compositions. Therefore, the present disclosure provides pharmaceutical compositions comprising a compound or a pharmaceutically acceptable salt of a Tdp1 inhibitor (such as a compound of Formula I) and at least one pharmaceutically acceptable carrier. The pharmaceutical composition / combination may contain a compound or salt of Formula I as the sole active agent, but preferably contains at least one additional active agent. In certain embodiments, it is preferred that the additional active agent is a compound or salt thereof selected from camptothecin, irinotecan and topotecan. In certain embodiments, the additional active agent is etoposide, teniposide, doxorubicin, daunorubicin, mitoxantrone, amsacrine, ellipticine, aurin tricarboxylic acid and 3-hydroxy-2-[(1R)-6-isopropenyl-3-methyl-cyclohex-2-ene-1-yl]-5-pentyl-1,4-benzoquinone (HU-331). In certain embodiments, the pharmaceutical composition is a dosage form containing about 0.1 mg to about 2000 mg, about 10 mg to about 1000 mg, about 100 mg to about 800 mg, or about 200 mg to about 600 mg of a compound of Formula I and optionally about 0.1 mg to about 2000 mg, about 10 mg to about 1000 mg, about 100 mg to about 800 mg, or about 200 mg to about 600 mg of an additional active agent in a unit dosage form. The pharmaceutical composition may also include a compound of a Tdp1 inhibitor (such as a compound of Formula I) and an additional active agent in a molar ratio. For example, the pharmaceutical composition may contain a TDP2 inhibitor of Formula I and an additional active agent in a molar ratio of about 0.5:1, about 1:1, about 2:1, about 3:1, or about 1.5:1 to about 4:1.

[0116] The compounds disclosed herein can be administered orally, topically, parenterally, by inhalation or spray, sublingually, transdermally, buccally, rectally, as an ophthalmic solution or otherwise in dosage unit formulations containing conventional pharmaceutically acceptable carriers. The pharmaceutical composition can be formulated into any pharmaceutically acceptable form, for example, as an aerosol, cream, gel, pill, capsule, tablet, syrup, transdermal patch or ophthalmic solution. Some dosage forms, such as tablets and capsules, are subdivided into unit doses of appropriate size containing an appropriate amount of active ingredient, such as an effective amount to achieve the desired purpose.

[0117] The carrier includes excipients and diluents, and must be of sufficiently high purity and sufficiently low toxicity to make it suitable for administration to the patient being treated. The carrier may be inert, or it may have its own pharmaceutical benefits. The amount of carrier used with the compound is sufficient to provide the actual amount of material for administration of the compound per unit dose.

[0118] Carrier categories include, but are not limited to, binders, buffers, colorants, diluents, disintegrants, emulsifiers, flavoring agents, glidants, lubricants, preservatives, stabilizers, surfactants, tableting agents, and wetting agents. Some carriers may be listed in more than one category, for example, vegetable oils may be used as lubricants in some formulations and as diluents in other formulations. Exemplary pharmaceutically acceptable carriers include sugars, starches, cellulose, powdered tragacanth, malt, gelatin; talc and vegetable oils. Optional active agents may be included in the pharmaceutical composition that do not substantially interfere with the activity of the compounds of the invention.

[0119] The pharmaceutical composition / combination can be formulated for oral administration. These compositions can contain 0.1 to 99 weight % (wt.%) of the compound of formula I, and typically at least about 5 wt.% of the compound of formula I. Some embodiments contain about 25 wt.% to about 50 wt.% or about 5 wt.% to about 75 wt.% of the compound of formula I.

[0120] Treatment

[0121] The compound of formula I or its salt and the pharmaceutical composition comprising the compound can be used to treat cancer, including causing tumor regression in vivo. The method of treating cancer or causing tumor regression includes providing a therapeutically effective amount of the compound of formula I to a patient in need of such treatment. In one embodiment, the patient is a mammal, more specifically a human. The disclosure also provides a method for treating non-human patients such as companion animals (e.g., cats, dogs and livestock). The therapeutically effective amount of the pharmaceutical composition can be an amount sufficient to inhibit the progression of cancer or cancerous tumors; or an amount that causes regression of cancer or cancerous tumors.

[0122] A therapeutically effective amount of a compound or pharmaceutical composition described herein will also provide a sufficient concentration of a compound of Formula I when administered to a patient. A sufficient concentration is the concentration of the compound necessary to prevent or resist a condition in the patient. Such an amount can be determined experimentally, for example, by measuring the blood concentration of the compound, or theoretically by calculating bioavailability.

[0123] The method of treatment includes providing a certain dosage of the compound of formula I to the patient. The dosage level of each compound of about 0.1 mg to about 140 mg per kilogram of body weight per day can be used to treat the above-mentioned conditions (about 0.5 mg to about 7 g per patient per day). The amount of the compound that can be combined with a carrier material to produce a single dosage form will vary depending on the patient and the specific mode of administration to be treated. The dosage unit form generally contains about 1 mg to about 500 mg of each active compound. In certain embodiments, a compound of formula I of 25 mg to 500 mg or 25 mg to 200 mg is provided to the patient every day. The frequency of dosage can also vary according to the compound used and the specific disease to be treated. However, in order to treat most diseases and conditions, a dosage regimen of 4 times a day or less can be used, and in certain embodiments, a dosage regimen of 1 or 2 times a day is used.

[0124] Formula I compounds can be used to treat cancer and cause tumor regression, including cancerous tumors. In certain embodiments, the patient suffers from a cell proliferative disorder or disease. Cell proliferative disorders can be cancer, tumors (cancerous or benign), tumors (neoplasm), neovascularization or melanoma. Cancers for treatment include solid cancers and disseminated cancers. Exemplary solid cancers (tumors) that can be treated by the methods provided herein include, for example, lung cancer, prostate cancer, breast cancer, liver cancer, colon cancer, breast cancer, kidney cancer, pancreatic cancer, brain cancer, skin cancer including malignant melanoma and Kaposi's sarcoma, testicular or ovarian cancer, cancer, sarcoma and kidney cancer (renal cell). Cancers that can be treated with Formula I compounds also include bladder cancer, breast cancer, colon cancer, endometrial cancer, lung cancer, bronchial cancer, melanoma, non-Hodgkin's lymphoma, blood cancer, pancreatic cancer, prostate cancer, thyroid cancer, brain or spinal cord cancer and leukemia. Exemplary disseminated cancers include leukemias or lymphomas, including Hodgkin's disease, multiple myeloma and mantle cell lymphoma (MCL), chronic lymphocytic leukemia (CLL), T-cell leukemia, multiple myeloma and Burkitt's lymphoma. Specifically included herein are methods of treating cancer by providing a compound of Formula I to a patient, wherein the cancer is a solid tumor or a disseminated cancer.

[0125] Further included are methods of treating cancer by providing a compound of Formula I to a patient, wherein the cancer is selected from glioma (glioblastoma, malignant glioma), acute myeloid leukemia, acute myeloid leukemia, myelodysplastic / myeloproliferative neoplasms, sarcoma, chronic myelomonocytic leukemia, non-Hodgkin's lymphoma, astrocytoma, melanoma, non-small cell lung cancer, small cell lung cancer, cervical cancer, rectal cancer, ovarian cancer, cholangiocarcinomas, chondrosarcoma or colon cancer.

[0126] However, it should be understood that the specific dosage level for any particular patient will depend on a variety of factors, including the activity of the specific compound used, age, body weight, general health, sex, diet, time of administration, route of administration and rate of excretion, drug combination and the severity of the specific disease being treated.

[0127] The compounds of Formula I can be administered alone (i.e., the sole therapeutic agent of a regimen) to treat or prevent diseases and conditions, such as undesirable cell proliferation, cancer and / or tumor growth, or can be administered in combination with another active agent. One or more compounds of Formula I can be administered in coordination with a regimen of one or more other chemotherapeutic agents, such as antineoplastic drugs, such as alkylating agents (e.g., nitrogen mustard, chlorambucil, cyclophosphamide, phenylalanine mustard, or ifosfamide), antimetabolites such as folic acid antagonists (e.g., methotrexate), purine antagonists (e.g., 6-mercaptopurine), or pyrimidine antagonists (e.g., 5-fluorouracil). Other non-limiting examples of chemotherapeutic agents that can be used in conjunction with one or more compounds of Formula I include taxanes and topoisomerase inhibitors. In addition, other non-limiting examples of active therapeutic agents include biologics, such as monoclonal antibodies or IgG chimeric molecules, which achieve their therapeutic effects by specifically binding to receptors or ligands in signal transduction pathways associated with cancer (e.g., therapeutic antibodies against CD20 (e.g., rituximab) or therapeutic antibodies against VEGF (e.g., bevacizumab)).

[0128] The treatment methods provided herein can also be used to treat mammals other than humans, including for veterinary applications, such as treating horses and livestock such as bulls, sheep, cows, goats, pigs, etc., and pets (companion animals) such as dogs and cats.

[0129] For diagnostic or research applications, a variety of mammals will be suitable subjects, including rodents (e.g., mice, rats, hamsters), rabbits, primates, and pigs such as inbred pigs, etc. In addition, for in vitro applications such as in vitro diagnostic and research applications, body fluids (e.g., blood, plasma, serum, interstitial fluid, saliva, feces, and urine) and cells and tissue samples of the above subjects will be suitable for use.

[0130] In one embodiment, the present invention provides a method of treating a cancer condition in a patient identified as in need of such treatment, the method comprising providing to the patient an effective amount of a compound of Formula I. The compounds or salts of Formula I provided herein can be administered alone or in combination with one or more other active agents.

[0131] In one embodiment, the method of treating cancer may additionally comprise determining that the cancer is responsive to Tdp2 inhibition.

[0132] In one embodiment, the method of treating cancer may additionally comprise administering to a patient in need of such treatment a compound of Formula I and one or more additional compounds, wherein at least one of the additional compounds is an agent known to be an inhibitor of topoisomerase 2.

[0133] In one embodiment, the method of treating cancer may additionally comprise administering to a patient in need of such treatment a therapeutically effective amount of a compound or salt of Formula I and one or more additional compounds, wherein at least one additional compound is an active agent selected from etoposide, teniposide, doxorubicin, daunorubicin, mitoxantrone, amsacrine, ellipticine, aurintricarboxylic acid and 3-hydroxy-2-[(1R)-6-isopropenyl-3-methyl-cyclohex-2-en-1-yl]-5-pentyl-1,4-benzoquinone (HU-331).

[0134] In other embodiments, the cancer is a cancer treatable using a Top2 inhibitor, wherein the TDP2 inhibitor as a combination therapy with an inhibitor of Top2 (eg, etoposide and doxorubicin) can provide a synergistic effect.

[0135] Example

[0136] abbreviation

[0137] DCM Dichloromethane

[0138] DMF N,N-Dimethylformamide

[0139] DMSO Dimethyl sulfoxide

[0140] DTT Dithiothreitol

[0141] EDTA Ethylenediaminetetraacetic acid

[0142] ESI Electrospray ionization

[0143] HPLC High Performance Liquid Chromatography

[0144] HRMS High Resolution Mass Spectrometry

[0145] NMR Nuclear Magnetic Resonance

[0146] THF Tetrahydrofuran

[0147] WCE whole cell extract

[0148] General approach

[0149] The reagents and solvents used were of commercial anhydrous grade. If not mentioned, they were used without further purification. Column chromatography was performed on silica gel (200-300 mesh). 1H NMR spectra were recorded on a Bruker AVANCE III 400 MHz spectrometer using tetramethylsilane as an internal reference. Mass spectra were analyzed on an Agilent 6120 (quadrupole LCMS) mass spectrometer. High resolution mass spectra were analyzed on a SHIMADZU LCMS-IT-TOF mass spectrometer. The HPLC analysis method used a SHIMADZU LC-20AB liquid chromatography system with a SPD-M20A detector. The analysis method conditions included a Phenomenex C18 column (4.6x250mm, 5.0um) and using NaHCO at pH 3.0 at a flow rate of 1 mL / min. 2 PO 4 Elution was performed with a linear gradient of 25% methanol to 75% methanol in aqueous buffer. Purity was determined using UV peak area at 220 nm. HPLC Method A used HCl with 0.1% TFA. 2 O buffer, method B used pH 6 phosphate-buffered saline buffer, and method C used pH 3 phosphate-buffered saline buffer.

[0150] Example

[0151] The following schemes provide general methods for preparing compounds of formula I,

[0152]

[0153] Example 1. Synthesis of Compounds 1 and 2

[0154]

[0155] To a yellow solution of 6,7-dichloroquinoline-5,8-dione (0.46 g, 2 mmol) in MeCN (30 ml) were added ethyl acetoacetate (0.26 ml, 2.2 mmol) and K 2 CO 3 (1.10 g, 8 mmol). The resulting solution was stirred and refluxed for 6 hours. After the reaction was completed, the reaction solution was cooled to room temperature and concentrated under reduced pressure. The target product was purified by silica gel column chromatography.

[0156] Ethyl 2-methyl-4,9-dioxo-4,9-dihydrofuro[3,2-g]quinoline-3-carboxylate (1), yellow solid. 1 HNMR (400MHz, CDCl 3)δ9.04(d,J=4.8Hz,1H),8.54(d,J=8.0Hz,1H),7.67(dd,J=7.6,4.7Hz,1H),4.4 6(q,J=7.1Hz,2H),2.76(s,3H),1.46(t,J=6.7Hz,3H).HRMS(ESI)m / z:284.0574 Calculated as C 15 H 10 NO 5 284.0564.

[0157] Ethyl 2-methyl-4,9-dioxo-4,9-dihydrofuro[2,3-g]quinoline-3-carboxylate (2), yellow solid. 1 HNMR (400MHz, CDCl 3 )δ9.06(d,J=4.0Hz,1H),8.54(d,J=7.6Hz,1H),7.70(dd,J=7.6,4.7Hz,1H),4.4 4(q,J=6.7Hz,2H),2.76(s,3H),1.48(t,J=7.2Hz,3H).HRMS(ESI)m / z:284.0551 Calculated as C 15 H 10 NO 5 284.0564.

[0158] Example 2. Synthesis of Compounds 8 and 9

[0159]

[0160] According to Preparation 1, acetylacetone was used as the starting material to obtain the target products 8 and 9.

[0161] 3-Acetyl-2-methylfuro[3,2-g]quinoline-4,9-dione (8), yellow solid. 1 H NMR (400 MHz, CDCl 3 )δ9.07(dd,J=4.7,1.7Hz,1H),8.54(dd,J=7.9,1.7Hz,1H),7.73(dd,J=7.9,4.7Hz,1H),2.79(s,3H),2.70(s,3H).HRMS(ESI)m / z:256.0593[M+H] + , calculated as C 14 H 10 NO 4 256.0604.

[0162] 3-Acetyl-2-methylfuro[2,3-g]quinoline-4,9-dione (9), yellow solid. 1 H NMR (400 MHz, CDCl 3 )δ9.07(d,J=4.5Hz,1H),8.55(dd,J=7.8,1.2Hz,1H),7.73(dd,J=7.8,4.7Hz,1H),2.81(s,3H),2.70(s,3H). HRMS(ESI)m / z:256.0613[M+H] + , calculated as C 14 H 10 NO 4 256.0604.

[0163] Example 3. Synthesis of Compound 3

[0164]

[0165] According to Preparation 1, using 2,3-dichloronaphthalene-1,4-dione as a starting material, the target product 3 (ethyl 2-methyl-4,9-dioxo-4,9-dihydronaphtho[2,3-b]furan-3-carboxylate) was obtained as a yellow solid. 1 H NMR (400 MHz, CDCl 3 )δ8.23-8.16(m,2H),7.76-7.74(m,2H),4.45(q,J=7.1Hz,1H),2.72(s,3H),1.45(t,J=7.1Hz,3H).HRMS(ESI)m / z:307.0590[M+Na] + , calculated as C 16 H 12 O 5 Na 307.0577.

[0166] Example 4. Synthesis of Compounds 29 and 30

[0167]

[0168] According to Preparation 1, using ethyl 3-oxo-3-phenylpropionate as a starting material, the target products 29 and 30 were obtained.

[0169] Ethyl 4,9-dioxo-2-phenyl-4,9-dihydrofuro[3,2-g]quinoline-3-carboxylate (29), yellow solid. 1 H NMR (400 MHz, CDCl 3)δ9.08(dd,J=4.6,1.6Hz,1H),8.53(dd,J=7.9,1.6Hz,1H),8.21-7.84(m,2H),7.72(dd,J=7.9,4.7Hz ,1H),7.60-7.42(m,3H),4.51(q,J=7.1Hz,2H),1.43(t,J=7.1Hz,3H).HRMS(ESI)m / z:348.0865[M+H] + , calculated as C 20 H 14 NO 5 348.0866.

[0170] Ethyl 4,9-dioxo-2-phenyl-4,9-dihydrofuro[2,3-g]quinoline-3-carboxylate (30), yellow solid. 1 H NMR (400 MHz, CDCl 3 )δ9.06(dd,J=4.7,1.7Hz,1H),8.58(dd,J=7.9,1.7Hz,1H),8.00-7.95(m,2H),7.72(dd,J=7.9,4.7Hz ,1H),7.52-7.49(m,3H),4.53(q,J=7.1Hz,2H),1.46(t,J=7.1Hz,3H).HRMS(ESI)m / z:348.0887[M+H] + , calculated as C 20 H 14 NO 5 348.0866.

[0171] Example 5. Preparation 2: Synthesis of Compound 7

[0172]

[0173] To a solution of 2 (1.10 g, 4 mmol) in MeOH or isopropanol (300 ml) was added K 2 CO 3 The reaction solution was refluxed for 12 hours. The precipitate was filtered out and dissolved in water (500 ml). The aqueous solution was acidified to pH 3 using hydrochloric acid (2N) and extracted with DCM (100 ml x 3). Anhydrous MgSO 4 The organic solution was dried and concentrated under reduced pressure. The resulting solid was recrystallized from MeOH to afford the target compound 7 as a yellow solid.

[0174] 2-Methyl-4,9-dioxo-4,9-dihydrofuro[2,3-g]quinoline-3-carboxylic acid (7). 1H NMR (400 MHz, CDCl 3 )δ9.12(dd,J=4.7,1.7Hz,1H),8.59(dd,J=7.9,1.7Hz,1H),7.82(dd,J=7.9,4.7Hz,1H),2.93(s,3H).

[0175] Example 6. Preparation 3: Synthesis of Compound 10

[0176]

[0177] To a red solution of compound 7 (65 mg, 0.25 mmol) and triethylamine (0.07 ml, 0.5 mmol) in freshly distilled chloroform (20 ml) was added dropwise thionyl chloride (1.25 ml) at room temperature. The mixture was stirred and refluxed for 5 hours. The reaction solution was then cooled to room temperature. The solvent was evaporated under reduced pressure. The residue was collected under reduced pressure for a period of time to remove most of the remaining thionyl chloride to obtain an orange solid residue. The obtained residue was dissolved in freshly distilled chloroform (10 ml) and added dropwise to a solution of 4-(dimethylamino)pyridine (35 mg, 0.3 mmol) and ethylene glycol (0.3 mmol) in freshly distilled chloroform (30 ml). The reaction mixture was refluxed for 3 hours and cooled to room temperature. The solvent was evaporated under reduced pressure. The target product was purified by silica gel column chromatography.

[0178] 2-Hydroxyethyl-2-methyl-4,9-dioxo-4,9-dihydrofuro[2,3-g]quinoline-3-carboxylate (10), yellow solid. 1 H NMR (400 MHz, CDCl 3 )δ9.08(d,J=3.2Hz,1H),8.56(d,J=7.6Hz,1H),7.73(dd,J=7.4,4.4Hz,1H),4.49( t,J=4.4Hz,2H),4.00(t,J=4,4Hz,2H),2.80(s,3H).HRMS(ESI)m / z:300.0499[MH] - , calculated as C 15 H 10 NO 6 300.0514.

[0179] Example 7. Synthesis of Compound 11

[0180]

[0181] According to Preparation 3, using propylene glycol as a starting material, the target yellow solid 11 was obtained.1 H NMR (400 MHz, CDCl 3 )δ9.06(d,J=3.3Hz,1H),8.54(d,J=7.0Hz,1H),7.71(dd,J=7.6,4.7Hz,1H),4.55(t,J=5.9Hz, 2H), 3.91 (t, J = 5.9Hz, 2H), 2.77 (s, 3H), 2.09 (quintet, J = 5.8Hz, 2H). HRMS (ESI) m / z: 314.0653 [MH] - , calculated as C 16 H 12 NO 6 314.0670.

[0182] Example 8. Synthesis of Compound 12

[0183]

[0184] According to Preparation 3, using m-dihydroxybenzene as a starting material, the target yellow solid 12 was obtained. 1 H NMR (400MHz, DMSO) δ9.81 (s, 1H), 9.03 (d, J = 3.1Hz, 1H), 8.49 (d, J = 7.2Hz, 1H), 7.87 (dd, J = 6.6 ,4.9Hz,1H),7.31-7.26(m,1H),6.88-6.68(m,3H),2.76(s,3H).HRMS(ESI)m / z:348.0499[MH] - , calculated as C 19 H 10 NO 6 348.0514.

[0185] Example 9. Synthesis of Compound 13

[0186]

[0187] According to Preparation 3, using o-dihydroxybenzene as a starting material, the target yellow solid 13 was obtained. 1 H NMR (400MHz, DMSO) δ9.81(s,1H),9.03(d,J=3.9Hz,1H),8.49(d,J=7.7Hz,1H),7.87(dd,J=7.6,4.7Hz,1H),7.23( d,J=7.8Hz,1H),7.16(t,J=7.6Hz,1H),7.00(d,J=7.9Hz,1H),6.90(t,J=7.5Hz,1H),2.80(s,3H).HRMS(ESI)m / z:C 19H 10 NO 6 348.0514[MH] - , calculated as 348.0512.

[0188] Example 10. Synthesis of Compound 14

[0189]

[0190] According to Preparation 3, using phenol as a starting material, the target yellow solid 14 was obtained. 1 H NMR (400MHz, DMSO) δ9.03 (dd, J=4.7, 1.7Hz, 1H), 8.49 (dd, J=7.9, 1.7Hz, 1H), 7.87 (dd, J=7.9 ,4.7Hz,1H),7.62-7.46(m,2H),7.44-7.29(m,3H),2.78(s,3H).HRMS(ESI)m / z:332.0563[MH] - , calculated as C 19 H 10 NO 5 332.0564.

[0191] Example 11. Synthesis of Compound 15

[0192]

[0193] According to Preparation 3, using phenol as a starting material, the target yellow solid 15 was obtained. 1 H NMR (400 MHz, CDCl 3 )δ9.08(dd,J=4.7,1.7Hz,1H),8.55(dd,J=7.9,1.7Hz,1H),7.73(dd,J=7.9,4.7Hz,1H),4.47(d ,J=5.2Hz,2H),4.19-4.17(m,1H),3.83-3.74(m,2H),2.80(s,3H).HRMS(ESI)m / z:330.0604[MH] - , calculated as C 16 H 12 NO 7 330.0619.

[0194] Example 12. Synthesis of Compound 16

[0195]

[0196] According to Preparation 3, using aniline as starting material, the target yellow solid 16 was obtained. 1H NMR (400MHz, DMSO) δ11.10(s,1H),9.06(d,J=3.2Hz,1H),8.51(dd,J=7.8,1.5Hz,1H),7.91(dd,J=7.7,4.7H z,1H),7.77-7.75(m,2H),7.44-7.40(m,2H),7.18-7.14(m,1H),2.78(s,3H).HRMS(ESI)m / z:331.0723[MH] - , calculated as C 19 H 11 N 2 O 4 331.0724.

[0197] Example 13. Synthesis of Compound 17

[0198]

[0199] According to Preparation 3, using m-chloroaniline as a starting material, the target yellow solid 17 was obtained. 1 H NMR (400MHz, DMSO) δ11.19(s,1H),9.07(d,J=3.2Hz,1H),8.51(dd,J=7.8,1.5Hz,1H),7.96(s,1H),7.92(dd,J=7.7,4.7 Hz,1H),7.59(d,J=8.2Hz,1H),7.45(t,J=8.1Hz,1H),7.23(d,J=7.8Hz,1H),2.76(s,3H).HRMS(ESI)m / z:367.0501[M+H] + , calculated as C 19 H 12 N 2 O 4 Cl 367.0480.

[0200] Example 14. Synthesis of Compound 18

[0201]

[0202] According to Preparation 3, using o-aminophenol as a starting material, the target yellow solid 18 was obtained. 1H NMR (400MHz, DMSO) δ11.11(s,1H),10.02(s,1H),9.05(d,J=3.3Hz,1H),8.49(d,J=6.7Hz,1H),8.18(d,J=7.8Hz,1H ),7.90(dd,J=7.7,4.7Hz,1H),7.01-6.93(m,2H),6.82(t,J=7.3Hz,1H),2.83(s,3H).HRMS(ESI)m / z:347.0656[MH] - , calculated as C 19 H 11 N 2 O 5 347.0673.

[0203] Example 15. Synthesis of Compound 19

[0204]

[0205] According to Preparation 3, using p-methoxyaniline as a starting material, the target red solid 19 was obtained. 1 H NMR (400MHz, DMSO) δ11.00(s,1H),9.07(d,J=3.3Hz,1H),8.50(dd,J=7.8,1.6Hz,1H),7.91(dd,J=7.7,4.7H z,1H),7.67(d,J=9.0Hz,2H),6.99(d,J=9.0Hz,2H),3.77(s,3H),2.77(s,3H).HRMS(ESI)m / z:361.0817[MH] - , calculated as C 20 H 13 N 2 O 5 361.0830.

[0206] Example 16. Synthesis of Compound 20

[0207]

[0208] According to Preparation 3, ethylamine was used as a starting material to obtain the target yellow solid 20. 1 H NMR (400 MHz, CDCl 3)δ9.52(s,1H),9.08(d,J=3.3Hz,1H),8.56(d,J=7.4Hz,1H),7.76(dd,J=7.7,4.7Hz,1H), 3.50(q,J=6.5Hz,2H),2.93(s,3H),1.34(t,J=7.2Hz,3H).HRMS(ESI)m / z:307.0699[M+Na] + , calculated as C 15 H 12 N 2 O 4 Na307.0689.

[0209] Example 17. Synthesis of Compound 22

[0210]

[0211] According to Preparation 3, using dimethyl 2-hydroxyethylphosphonate as a starting material, the target yellow solid 22 was obtained. 1 H NMR (400 MHz, CDCl 3 )δ9.06(dd,J=4.6,1.6Hz,1H),8.54(dd,J=7.9,1.6Hz,1H),7.71(dd,J=7.9,4.7Hz,1H),4.64-4.5 8(m,2H),3.80(s,3H),3.77(s,3H),2.77(s,3H),2.54–2.39(m,2H).HRMS(ESI)m / z:392.0538[MH] - , calculated as C 17 H 15 NO 8 P 392.0541.

[0212] Example 18. Synthesis of Compound 23

[0213]

[0214] According to Preparation 3, using m-chlorophenol as the starting material, the target yellow solid 23 was obtained. 1 H NMR (400 MHz, CDCl 3 )δ9.07(dd,J=4.6,1.6Hz,1H),8.56(dd,J=7.9,1.6Hz,1H),7.72(dd,J=7.9,4.7Hz,1H),7.5 2(s,1H),7.39-7.35(m,2H),7.30-7.26(m,1H),2.83(s,1H).HRMS(ESI)m / z:368.0341[M+H] + , calculated as C19 H 11 NO 5 Cl 368.0320.

[0215] Example 19. Synthesis of Compound 24

[0216]

[0217] According to Preparation 3, using o-chlorophenol as starting material, the target yellow solid 24 was obtained. 1 H NMR (400 MHz, CDCl 3 )δ9.06(dd,J=4.6,1.6Hz,1H),8.56(dd,J=7.9,1.7Hz,1H),7.72(dd,J=7.9,4.7Hz,1H),7.53-7.49 (m,2H),7.38(td,J=7.8,1.5Hz,1H),7.29-7.25(m,1H),2.85(s,3H).HRMS(ESI)m / z:368.0335[M+H] + , calculated as C 19 H 10 NO 5 Cl 368.0320.

[0218] Example 20. Synthesis of Compound 25

[0219]

[0220] According to Preparation 3, using 2-naphthol as starting material, the target yellow solid 25 was obtained. 1 H NMR (400 MHz, CDCl 3 )δ9.07(d,J=4.6Hz,1H),8.56(d,J=7.8Hz,1H),7.94-9.86(m,4H),7.72(dd,J=7.9,4.6Hz,1H ),7.60(dd,J=8.8,2.2Hz,1H),7.55-7.41(m,2H),2.86(s,3H).HRMS(ESI)m / z:382.0706[MH] - , calculated as C 23 H 12 NO 5 382.0721.

[0221] Example 21. Synthesis of Compound 26

[0222]

[0223] According to Preparation 3, using 3-hydroxybenzonitrile as the starting material, the target yellow solid 26 was obtained.1 H NMR (400 MHz, CDCl 3 )δ9.08(dd,J=4.7,1.7Hz,1H),8.57(dd,J=7.9,1.7Hz,1H),7.82(s,1H),7.79-7.75(m,1H), 7.74(dd,J=7.9,4.7Hz,1H),7.66-7.48(m,2H),2.84(s,3H).HRMS(ESI)m / z:359.0679[M+H] + , calculated as C 20 H 11 N 2 O 5 359.0662.

[0224] Example 22. Synthesis of Compound 27

[0225]

[0226] According to Preparation 3, using p-chlorophenol as the starting material, the target yellow solid 27 was obtained. 1 H NMR (400 MHz, CDCl 3 )δ9.07(dd,J=4.6,1.6Hz,1H),8.56(dd,J=7.9,1.6Hz,1H),7.72(dd,J=7.9,4.7Hz,1H),7.41(br,s,4H),2.82(s,3H).HRMS(ESI)m / z:368.0334[M+H] + , calculated as C 19 H 11 NO 5 Cl368.0320.

[0227] Example 23. Synthesis of Compound 28

[0228]

[0229] According to Preparation 3, using 3-hydroxypyridine as a starting material, the target yellow solid 28 was obtained. 1 H NMR (400 MHz, CDCl 3 )δ9.08(dd,J=4.7,1.7Hz,1H),8.77(d,J=2.5Hz,1H),8.58-8.55(m,2H),7.87(ddd,J=8.3,2.7,1.4Hz,1 H),7.73(dd,J=7.9,4.7Hz,1H),7.43(dd,J=8.3,4.8Hz,1H),2.85(s,3H).HRMS(ESI)m / z:335.0673[M+H]+ , calculated as C 18 H 11 N 2 O 5 335.0662.

[0230] Example 24. Synthesis of Compound 31

[0231]

[0232] According to Preparation 3, using 4-bromophenol as a starting material, the target yellow solid 31 was obtained. 1 H NMR (400 MHz, CDCl 3 )δ9.07(dd,J=4.7,1.7Hz,1H),8.56(dd,J=7.9,1.7Hz,1H),7.72(dd,J=7.9,4.7Hz,1H),7.56( d,J=8.8Hz,2H),7.36(d,J=8.8Hz,2H),2.82(s,3H).HRMS(ESI)m / z:411.9820 and 413.9865[M+H] + , calculated as C 19 H 11 NO 5 BR 411.9815 and 413.9797.

[0233] Example 25. Synthesis of Compound 32

[0234]

[0235] According to Preparation Example 3, 3-ethynylphenol was used as a raw material to obtain the target yellow solid 32. 1 H NMR (400 MHz, CDCl 3 )δ9.07(dd,J=4.6,1.5Hz,1H),8.56(dd,J=7.9,1.7Hz,1H),7.72(dd,J=7.9,4.7Hz,1H),7 .60(s,1H),7.50-7.34(m,3H),3.12(s,1H),2.83(s,3H).HRMS(ESI)m / z:380.0543[M+Na] + , calculated as C 21 H 11 NO 5 Na 380.0529.

[0236] Example 26. Synthesis of Compound 35

[0237]

[0238] According to Preparation Example 3, butane-1,4-diol was used as a raw material to obtain the target yellow solid 35. 1 H NMR (400MHz, CDCl3) δ9.06 (d, J=3.5Hz, 1H), 8.54 (dd, J=7.8, 1.1Hz, 1H), 7.71 (dd, J=7.8, 4.6Hz, 1H), 4.43 (t, J=6. 4Hz,2H),3.75(t,J=6.3Hz,2H),2.76(s,3H),2.00-1.92(m,2H),1.85-1.78(m,2H).HRMS(ESI)m / z:352.0810[M+Na] + , calculated as C 17 H 15 NO 6 Na 352.0792.

[0239] Example 27. Synthesis of Compound 36

[0240]

[0241] According to Preparation 3, using 2,2'-oxydiethanol as a starting material, the target yellow solid 36 was obtained. 1 H NMR (400 MHz, CDCl 3 )δ9.05(dd,J=4.7,1.7Hz,1H),8.53(dd,J=7.9,1.7Hz,1H),7.71(dd,J=7.9,4.7Hz,1H),4.54(t,J=4.6Hz,2H),3 .93(t,J=4.8Hz,2H),3.79(t,J=4.4Hz,2H),3.70(t,J=4.4Hz,2H),2.76(s,3H).HRMS(ESI)m / z:368.0752[M+Na] + , calculated as C 17 H 15 NO 7 Na368.0741.

[0242] Example 28. Synthesis of Compound 37

[0243]

[0244] According to Preparation 3, using (R)-(2,2-dimethyl-1,3-dioxolan-4-yl)methanol as a starting material, the target yellow solid 37 was obtained. 1 H NMR (400 MHz, CDCl 3)δ9.09(dd,J=4.6,1.6Hz,1H),8.57(dd,J=7.8,1.5Hz,1H),7.75(dd,J=7.8,4 .7Hz,1H),4.48(d,J=4.7Hz,2H),4.19(m,1H),3.85-3.76(m,2H),2.81(s,3H).

[0245] Example 29. Synthesis of Compounds 38 and 45

[0246]

[0247] According to Preparation 3, using (S)-(2,2-dimethyl-1,3-dioxolan-4-yl)methanol as starting material, two yellow solids 38 and 45 were obtained.

[0248] 38: 1 H NMR (400 MHz, CDCl 3 )δ8.99(dd,J=4.7,1.7Hz,1H),8.47(dd,J=7.9,1.7Hz,1H),7.64(dd,J=7.9,4.7Hz,1H),4.54-4.46(m,1H),4.36(dd,J=11.0,5.4Hz,1H),4.30(dd,J =11.0,6.5Hz,1H),4.19(dd,J=8.7,6.3Hz,1H),3.94(dd,J=8.7,5.4Hz,1H),2.70(s,3H),1.37(s,3H),1.31(s,3H).HRMS(ESI)m / z:394.0909[M+Na] + , calculated as C 19 H 17 NO 7 Na394.0897.

[0249] 45: 1 H NMR (400 MHz, CDCl 3 )δ9.01(dd,J=4.7,1.7Hz,1H),8.48(dd,J=7.9,1.7Hz,1H),7.66(dd,J=7.9,4.7Hz,1H),4.40(d, J=5.2Hz,2H),4.14-4.08(m,1H),3.76-3.63(m,2H),2.73(s,3H).HRMS(ESI)m / z:354.0596[M+Na] + , calculated as C 16 H 13 NO 7 Na 354.0584.

[0250] Example 30. Synthesis of Compound 39

[0251]

[0252] According to Preparation 3, using benzyl alcohol as starting material, the target yellow solid 39 was obtained. 1 H NMR (400MHz, CDCl3) δ9.06 (dd, J=4.6, 1.6Hz, 1H), 8.53 (dd, J=7.9, 1.6Hz, 1H), 7.69 (dd, J=7.8, 4.7Hz, 1H), 7.5 8-7.56(m,2H),7.42-7.37(m,2H),7.35-7.31(m,1H),5.42(s,2H),2.73(s,3H).HRMS(ESI)m / z:370.0704[M+Na] + , calculated as C 20 H 13 NO 5 Na 370.0686.

[0253] Example 31. Synthesis of Compound 40

[0254]

[0255] According to Preparation 3, using 2-phenylethanol as a starting material, the target yellow solid 40 was obtained. 1 H NMR(400MHz, CDCl3) δ9.07(dd,J=4.7,1.7Hz,1H),8.54(dd,J=7.9,1.7Hz,1H),7.70(dd,J=7.9,4.7Hz,1H),7.33-7.28 (m,4H),7.25-7.20(m,1H),4.60(t,J=7.3Hz,2H),3.19(t,J=7.3Hz,2H),2.65(s,3H).HRMS(ESI)m / z:384.0864[M+Na] + , calculated as C 21 H 15 NO 5 Na 384.0842.

[0256] Example 32. Synthesis of Compound 58

[0257]

[0258] According to Preparation Example 3, pentane-1,5-diol was used as a raw material to obtain the target yellow solid 45. 1 H NMR (400 MHz, CDCl 3)δ9.05(dd,J=4.7,1.7Hz,1H),8.53(dd,J=7.9,1.7Hz,1H),7.70(dd,J=7.9,4.7Hz,1H),4.40(t,J=6.5Hz,2H ),3.71(t,J=6.1Hz,2H),2.76(s,3H),1.93-1.85(m,2H),1.73-1.55(m,4H).HRMS(ESI)m / z:366.0966[M+Na] + , calculated as C 18 H 17 NO 6 Na 366.0948.

[0259] Example 33. Synthesis of Compound 46

[0260]

[0261] According to Preparation 3, using 2-hydroxyacetic acid as starting material, the target yellow solid 46 was obtained. 1 H NMR(400MHz,MeOD)δ8.95(dd,J=4.7,1.6Hz,1H),8.58(dd,J=7.9,1.6Hz,1H),7.84 (dd,J=7.9,4.8Hz,1H),4.07(s,2H),2.78(s,3H).HRMS(ESI)m / z:338.0283[M+Na] + , calculated as C 15 H 9 NO 7 Na338.0271.

[0262] Example 34. Synthesis of Compound 47

[0263]

[0264] According to Preparation 3, using trifluoroethanol as starting material, the target yellow solid 47 was obtained. 1 H NMR (400 MHz, CDCl 3 )δ9.07(dd,J=4.7,1.7Hz,1H),8.54(dd,J=7.9,1.7Hz,1H),7.71(dd,J=7.9,4 .7Hz,1H),4.77(q,J=8.3Hz,2H),2.78(s,3H).HRMS(ESI)m / z:362.0266[M+Na] + , calculated as C 15 H 8 NO 5 F 3Na362.0247.

[0265] Example 35. Synthesis of Compound 48

[0266]

[0267] According to Preparation 3, using 3-hydroxypropionitrile as starting material, the target yellow solid 48 was obtained. 1 H NMR (400 MHz, CDCl 3 )δ9.06(dd,J=4.6,1.6Hz,1H),8.54(dd,J=7.9,1.7Hz,1H),7.72(dd,J=7.9,4.7Hz,1H),4.60(t,J=6.5Hz,2H),2.99(t,J=6.5Hz,2H),2.78(s,3H).

[0268] Example 36. Synthesis of Compound 49

[0269]

[0270] According to Preparation 3, benzo[d][1,3]dioxol-5-ol was used as the starting material to obtain the target yellow solid 49. 1 H NMR (400 MHz, CDCl 3 )δ9.06(dd,J=4.7,1.7Hz,1H),8.55(dd,J=7.9,1.7Hz,1H),7.72(dd,J=7.9,4.7Hz,1H),6.97( d,J=2.3Hz,1H),6.89(dd,J=8.4,2.3Hz,1H),6.82(d,J=8.4Hz,1H),6.01(s,2H),2.81(s,2H).

[0271] Example 37. Synthesis of Compound 43

[0272]

[0273] According to Preparation Example 3, prop-2-yn-1-ol was used as a raw material to obtain the target yellow solid 43. 1 H NMR (400 MHz, CDCl 3 )δ9.06(d,J=4.5Hz,1H),8.54(d,J=7.7Hz,1H),7.70(dd,J=7.8,4.6Hz,1H),4.50(t,J =7.0Hz,2H),2.88-2.64(m,5H),2.02(t,J=2.6Hz,1H).HRMS(ESI)m / z:332.0544[M+Na]+ , calculated as C 17 H 11 NO 5 Na 332.0529.

[0274] Example 38. Synthesis of Compound 44

[0275]

[0276] According to Preparation 3, using 4-(methylsulfonyl)phenol as a starting material, the target yellow solid 44 was obtained. 1 H NMR (400 MHz, CDCl 3 )δ9.10(dd,J=4.7,1.7Hz,1H),8.59(dd,J=7.9,1.7Hz,1H),8.11-8.03(m,2H),7.76(dd,J=7 .9,4.7Hz,1H),7.76-7.63(m,2H),3.11(s,3H),2.86(s,3H).HRMS(ESI)m / z:434.0334[M+H] + , calculated as C 20 H 13 NO 7 SNa 434.0305.

[0277] Example 39. Synthesis of Compound 50

[0278]

[0279] According to Preparation 3, using 2-hydroxybenzonitrile as a starting material, the target yellow solid 50 was obtained. 1 H NMR (400 MHz, CDCl 3 )δ9.06(dd,J=4.7,1.7Hz,1H),8.57(dd,J=7.9,1.7Hz,1H),7.92-7.61(m,4H ),7.42(td,J=7.5,1.4Hz,1H),2.87(s,3H).HRMS(ESI)m / z:381.0503[M+Na] + , calculated as C 20 H 10 N 2 O 5 Na381.0482.

[0280] Example 40. Synthesis of Compound 51

[0281]

[0282] According to Preparation 3, using 4-hydroxybenzonitrile as starting material, the target yellow solid 51 was obtained. 1 H NMR (400 MHz, CDCl 3 )δ9.08(dd,J=4.7,1.7Hz,1H),8.57(dd,J=7.9,1.7Hz,1H),7.76(d,J=8.8Hz,1H),7.74(d d,J=7.0,3.7Hz,2H),7.63(d,J=8.8Hz,2H),2.84(s,3H).HRMS(ESI)m / z:381.0496[M+Na] + , calculated as C 20 H 10 N 2 O 5 Na 381.0482.

[0283] Example 41. Synthesis of Compound 52

[0284]

[0285] According to Preparation 3, using N-acetyl-L-tyrosine methyl ester as the material, the target yellow solid 52 was obtained. 1 HNMR (400MHz, CDCl 3 )δ9.06(dd,J=4.7,1.7Hz,1H),8.56(dd,J=7.9,1.7Hz,1H),7.72(dd,J=7.9,4.7Hz,1H),7.38(d,J=8.5Hz,2H),7.17(d,J=8.5Hz,2 H),5.98(d,J=7.5Hz,1H),4.91(dt,J=7.7,5.7Hz,1H),3.75(s,3H),3.17(dd,J=5.6,3.4Hz,2H),2.82(s,3H),2.02(s,3H).ESI / MS m / z:477.1[M+H] + .

[0286] Example 42. Synthesis of Compounds 53 and 54

[0287]

[0288] According to Preparation 3, using 3-bromopropanol as starting material, the target yellow solid 53 was obtained. 1 H NMR (400 MHz, CDCl 3)δ9.06(dd,J=4.7,1.7Hz,1H),8.54(dd,J=7.9,1.7Hz,1H),7.70(dd,J=7.9,4.7Hz,1H),4.54(t,J=5.8H z,2H),3.75(t,J=5.4Hz,2H),2.78(s,3H),2.45-2.36(m,2H).HRMS(ESI)m / z:377.9996 and 380.0005[M+H] + , calculated as C 16 H 12 NO 5 BR 377.9972 and 379.9953.

[0289] A solution of 53 (20 mg, 0.05 mmol) in pyridine (5 ml) was heated at 80°C for 8 hours. The reaction solution was then cooled to room temperature and ether (5 ml) was added. The precipitate was filtered and washed with ether (10 ml) and ethyl acetate (5 ml) to obtain the target yellow solid 54. 1 H NMR(400MHz,MeOD)δ9.20(d,J=5.6Hz,2H),9.02(dd,J=4.7,1.6Hz,1H),8.64-8.60(m,2H),8.18(t,J=7.2Hz,2H),7.89(dd,J= 7.9,4.8Hz,1H),5.08(t,J=7.4Hz,2H),4.46(t,J=5.6Hz,2H),2.74(s,3H),2.64-2.48(m,2H).HRMS(ESI)m / z:377.1149[M-Br] + , calculated as C 21 H 17 N 2 O 5 377.1132.

[0290] Example 43. Synthesis of Compound 55

[0291]

[0292] A solution of 53 (20 mg, 0.05 mmol) and N,N-dimethyl-4-aminopyridine (62 mg, 0.5 mmol) in THF (10 ml) was refluxed for 8 hours. The reaction solution was then cooled to room temperature and ether (20 ml) was added. The precipitate was filtered out and washed with ether (10 ml) and ethyl acetate (5 ml) to obtain the target yellow solid 55. 1H NMR (400MHz, MeOD) δ9.01(dd,J=4.7,1.5Hz,1H),8.61(dd,J=7.9,1.5Hz,1H),8.35(d,J=7.7Hz,2H),7.89(dd,J=7.9,4.8Hz,1H),7.01(d, J=7.7Hz,2H),4.57(t,J=7.0Hz,2H),4.40(t,J=5.6Hz,2H),3.21(s,6H),2.74(s,3H),2.45-2.31(m,2H).HRMS(ESI)m / z:420.1555[M-Br] + , calculated as C 23 H 22 N 3 O 5 420.1554.

[0293] Example 44. Preparation 4: Synthesis of Compound 33

[0294]

[0295] To a solution of 3-bromopropionic acid (152 mg, 1 mmol) in MeCN (20 ml) was added NaN 3 (130 mg, 2 mmol). The solution was refluxed for 8 hours. The reaction solution was concentrated under reduced pressure to obtain a white solid. The obtained white solid was dissolved in water (5 ml), and compound 32 (60 mg, 0.17 mmol), CuSO 4 (3 mg), sodium ascorbate (2.5 mg) and DMF (5 ml). The solution was heated to 80°C for 3 hours. Then, the reaction solution was cooled to room temperature, and water (50 ml) was added. CH 2 Cl 2 The resulting suspension was extracted with 1% ethyl acetate (20 ml x 2). The combined organic solution was washed with water (10 ml x 3) and saturated brine solution (10 ml x 2). The organic solution was concentrated under reduced pressure and purified by silica gel column chromatography to obtain the target yellow solid 33. 1H NMR (400MHz, DMSO) δ12.54 (s, 1H), 9.04 (dd, J = 4.6, 1.6Hz, 1H), 8.68 (s, 1H), 8. 50(dd,J=7.9,1.6Hz,1H),7.89(dd,J=7.9,4.7Hz,1H),7.85(d,J=1.9Hz,1H),7. 82(d,J=7.8Hz,1H),7.60(t,J=7.9Hz,1H),7.35(dd,J=8.1,1.4Hz,1H),4.63(t, J=6.7Hz,2H),2.97(t,J=6.7Hz,2H),2.81(s,3H).HRMS(ESI)m / z:471.0920[MH] - , calculated as C 24 H 15 N 4 O 7 471.0946.

[0296] Example 45. Synthesis of Compound 34

[0297]

[0298] According to Preparation 4, using 3-bromopropanol as starting material, the target yellow solid 34 was obtained. 1 H NMR (400 MHz, CDCl 3 )δ9.08(dd,J=4.7,1.7Hz,1H),8.58(dd,J=7.9,1.7Hz,1H),7.93(s,1H),7.85-7.82(m,2H),7.74(dd,J=7.9,4.7Hz,1H),7.52(t,J=7.9Hz,1H),7.44-7.37(m,1H),4.61(t,J=6.7Hz,2H),3.72(t,J=5.8Hz,2H),2.85(s,3H),2.21(quintet,J=6.2Hz,2H).HRMS(ESI)m / z:457.1138[MH] - , calculated as C 24 H 17 N 4 O 6 457.1154.

[0299] Example 46. Synthesis of Compound 42

[0300]

[0301] According to Preparation 4, using compound 43 as a starting material, the target yellow solid 42 was obtained. 1H NMR (400 MHz, CDCl 3 )δ8.99(dd,J=4.8,1.4Hz,1H),8.65(dd,J=7.9,1.5Hz,1H),8.19(s,1H),7.83(dd,J=7.9,4.8Hz,1H),4.78(t,J=5.6H z,2H),4.70(t,J=5.0Hz,2H),3.24(t,J=5.0Hz,2H),2.92(t,J=5.6Hz,2H),2.79(s,2H).HRMS(ESI)m / z:423.0923[MH] - , calculated as C 20 H 15 N 4 O 7 423.0946.

[0302] Example 47. Synthesis of Compound 56

[0303]

[0304] According to the synthesis of 42, using 3-bromopropanol as a starting material, the target yellow solid 56 was obtained. 1 H NMR (400 MHz, CDCl 3 )δ9.02(d,J=4.5Hz,1H),8.57(dd,J=7.9,1.6Hz,1H),7.98(s,1H),7.75(dd,J=7.9,4.7Hz,1H),4.70(t,J=5.6Hz,2H),4.58(t, J=5.6Hz,2H),3.68(t,J=5.8Hz,2H),3.27(t,J=5.6Hz,2H),2.76(s,3H),2.15(quintet,J=6Hz,2H).HRMS(ESI)m / z:433.1142[M+Na] + , calculated as C 20 H 18 N 4 O 6 Na 433.1119.

[0305] Example 48. Synthesis of Compound 7-Na Salt

[0306]

[0307] To a solution of 7 (52 mg, 0.20 mmol) in ethanol (10 ml), a solution of NaOH (8 mg, 0.2 mmol) in ethanol (5 ml) was added dropwise. The solution was stirred at room temperature for 30 minutes. The precipitate was filtered out, washed with ethanol (1 ml x 2) and dried to give the target 7-Na salt as a yellow solid.

[0308] Example 49. Synthesis of Compound 33-Na Salt

[0309]

[0310] According to the synthesis of 7-Na salt, compound 33 was used as a raw material to obtain the target yellow solid 33-Na salt.

[0311] Example 50. Synthesis of Compound 57

[0312]

[0313] 2 (72 mg, 0.25 mmol), hydroxylamine hydrochloride (52 mg, 0.75 mmol) and K were mixed in ethanol (10 ml). 2 CO 3 The solution of (52 mg, 0.375 mmol) was stirred and refluxed for 1.5 h. The reaction solution was concentrated under reduced pressure. The obtained solid was purified by silica gel column chromatography to obtain the target yellow solid 57. 1 H NMR (400 MHz, CDCl 3 )δ8.76-8.74(m,2H),7.76(dd,J=8.0,5.0Hz,1H),4.42(q,J=7.1Hz,2H),2.64(s,3H),1.41(t,J=7.1Hz,3H).

[0314] Example 51. Additional compounds

[0315] The following compounds can be prepared by the methods shown in Examples 1 to 50. Routine variations in reagents and reaction conditions required to prepare a particular compound will be apparent to those skilled in the art.

[0316]

[0317] Example 52. Whole Cell Extract TDP1 Assay

[0318] DT40 knockout cells (1x10 7). The cell pellet was then resuspended in 100 μL CellLytic M cell lysis reagent (SIGMA-Aldrich C2978). After 15 min on ice, the lysate was centrifuged at 12,000 g for 10 min, and the supernatant was transferred to a fresh tube. Protein concentration was determined using a Nanodrop spectrophotometer (Invitrogen), and whole-cell extracts were stored at -80 °C. 5′-[ 32 The single-stranded DNA oligonucleotide of [P]-labeling, at room temperature, in the WCE buffer containing 50mM Tris HCl, pH 7.5,80mM KCl, 2mM EDTA, 1mM DTT, 40μg / mL BSA and 0.01% Tween-20, under the condition of the presence or absence of inhibitor, use 4μg / mL whole cell extract to incubate 15 minutes under 1nM.By adding 1 volume of gel loading buffer [99.5% (v / v) formamide, 5mM EDTA, 0.01% (w / v) xylene cyanol (xylene cyanol) and 0.01% (w / v) bromophenol blue] termination reaction.With 12 minutes interval, sample is carried out to 16% denaturation PAGE with multiple loading.Gel is dried and exposed to PhosphorImager screen (GE Healthcare).Use Typhoon 8600 (GE Healthcare) to scan gel image, and use ImageQuant software (GE Healthcare) to carry out optical density analysis.

[0319] Example 53. Recombinant TDP1 Assay

[0320] N14Y DNA substrate was incubated at 1 nM with 10 pM recombinant TDP1 for 15 minutes at room temperature in WCE buffer (see Example 51) in the absence or presence of inhibitors. When indicated, parallel reactions were performed in HTS assay buffer containing 1X PBS, pH 7.4, 80 mM KCl, and 0.01% Tween-20. Samples were then analyzed similarly to the WCE TDP1 assay (Example 51).

[0321] Example 54. Recombinant TDP2 Assay

[0322] The TDP2 reaction was performed as previously described (Example 52) with the following modifications. The reaction was carried out at room temperature in a flask containing 50 mM Tris-HCl, pH 7.5, 80 mM KCl, 5 mM MgCl 2, 0.1 mM EDTA, 1 mM DTT, 40 μg / mL BSA and 0.01% Tween 20 buffer, in the absence or presence of inhibitors, 25 pM recombinant human TDP2 was used to react with an 18-mer single-stranded oligonucleotide DNA substrate (α 32 P-cordycepin-3'-labeled) was incubated for 15 minutes. The reaction was terminated and processed similarly to the WCE and recombinant TDP1 reactions (see Examples 52 and 53).

[0323] Example 55. Whole Cell Extract TDP2 Assay

[0324] DT40 knockout cells (1x10 7 The cell pellets were then lysed and stored similarly to the hTDP1 extract (see above). 2 , 2mM EDTA, 1mM DTT, 40μg / mL BSA and 0.01% Tween-20 in WCE buffer, in the absence or presence of inhibitors, 4μg / mL whole cell extract was used to incubate the 18-mer single-stranded oligonucleotide at 1nM for 15 minutes. The reaction was terminated by adding 1 volume of gel loading buffer [99.5% (v / v) formamide, 5mM EDTA, 0.01% (w / v) xylene cyanol and 0.01% (w / v) bromophenol blue]. The samples were then processed and run similarly to the hTDP1 whole cell extract (Example 51).

[0325] Example 56. Additional compounds

[0326] Table 1 shows the compounds of compounds 1 to 44 and additional compounds 59-74. All compounds were prepared by the methods shown in Examples 1 to 50. The specific compounds disclosed in Table 1 were prepared using routine variations of starting materials and reaction conditions that would be apparent to one skilled in the art. 4 stars "****" are used to indicate IC 50 Compounds ≤ 1 micromolar, 3 stars "***" indicates 1 micromolar <IC 50 Compounds with ≤12 micromolar, 2 stars "**" indicates 12 micromolar <IC 50 Compounds with ≤37 μmol, one star “*” indicates 37 μmol <IC 50 ≤111 μmol of compound, and 0 indicates an IC 50>111 micromolar compounds. Use a standard Tdp2 inhibition assay, such as the assay of Example 53, to determine the IC of the compound. 50 . LC methods are given in the "General Methods" section.

[0327]

[0328]

[0329]

[0330]

[0331]

[0332]

[0333]

[0334]

[0335]

[0336]

[0337]

[0338]

Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof, in X 1 Is N or CR 5 ; X 2 Is N or CR 6 ;and X 1 or X 2 At least one of them is N; R 1 It is ethoxy; Y 1 It is O; Y 2 Is N or CR 2 ; R 2 is C1-C6 haloalkyl, -(C0-C6 alkyl)NR 7 R 8 or phenyl, each R except halogen 2 is substituted at each occurrence with 0 to 3 groups independently selected from halogen, hydroxy, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl and C1-C6 haloalkoxy; R 3 , R 4 , R 5 and R 6 is hydrogen; and Any R attached to the same nitrogen atom 7 and R 8 Together they form a 4- to 7-membered heterocycloalkyl group substituted with 0 to 2 substituents selected from the group consisting of hydroxy, halogen, C1-C4 alkyl, C1-C4 alkoxy and C2-C4 alkanoyl.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Y 2 It is N.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Y 1 is O and Y 2 It is N.

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Y 2 is N; and R 2 is C1-C6 haloalkyl or phenyl, said phenyl being substituted at each occurrence by 0 to 3 groups independently selected from halogen, hydroxy, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl and C1-C6 haloalkoxy.

5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Y 2 Yes CR 2 And R 2 It is a C1-C6 haloalkyl group.

6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is 7. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is 8. A pharmaceutical composition comprising the compound according to claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

9. Use of a compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating cancer in a patient in need of such treatment.

10. Use of a compound as claimed in any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, in combination with one or more additional compounds in the manufacture of a medicament for treating cancer in a patient in need of such treatment, wherein at least one of the additional compounds is an active agent known as an inhibitor of topoisomerase 2.

11. The use according to claim 10, wherein at least one of the additional compounds is an active agent selected from etoposide, teniposide, doxorubicin, daunorubicin, mitoxantrone, amsacrine, ellipticine, aurintricarboxylic acid and 3-hydroxy-2-[(1R)-6-isopropenyl-3-methyl-cyclohex-2-en-1-yl]-5-pentyl-1,4-benzoquinone.

12. The method of any one of claims 9 to 11, wherein the cancer is glioma, acute myeloid leukemia, acute myeloid leukemia, myelodysplastic / myeloproliferative neoplasm, sarcoma, chronic myelomonocytic leukemia, non-Hodgkin's lymphoma, astrocytoma, melanoma, non-small cell lung cancer, small cell lung cancer, cervical cancer, rectal cancer, ovarian cancer, bile duct cancer, chondrosarcoma or colon cancer.

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