Tricyclic phthalazinone PARP inhibitors and methods of use

Abstract

The present disclosure relates to a compound # imgabs0 # according to Formula I or a pharmaceutically acceptable salt and / or solvate thereof, as well as compositions comprising such compounds and uses thereof, where X1 is H, F or Cl; x2 is NH or N-R3; one of R1 and R2 is H, halo, alkyl, alkenyl, alkynyl, cycloalkyl, or heterocyclic group, and the remaining one of R1 and R2 is aryl, heteroaryl, or non-aromatic heterocyclic group; and R3 is an alkyl group, a cycloalkyl group, an alkylene group, or a non-aromatic heterocyclic group. In addition, the present disclosure demonstrates that the compounds of the present disclosure penetrate the central nervous system, thereby allowing the treatment of central nervous system cancers.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of and priority to Provisional Patent Application No. 63 / 350,366, filed on June 8, 2022, which is incorporated herein by reference in its entirety for any and all purposes. Technical Field

[0003] The present technology relates to compounds, compositions and methods related to the treatment of cancer, especially central nervous system cancer and their uses. Summary of the invention

[0004] In one aspect, the present technology provides a compound according to Formula I

[0005]

[0006] or a pharmaceutically acceptable salt and / or solvate thereof, wherein X 1 is H, F or Cl; X 2 Is NH or NR 3 ; R 1 and R 2 One of is H, halo, alkyl, alkenyl, alkynyl, cycloalkyl or heterocyclyl, and R 1 and R 2 The other one of R is an aryl group, a heteroaryl group or a non-aromatic heterocyclic group; 3 It is an alkyl group, a cycloalkyl group, an alkylene group and a non-aromatic heterocyclic group.

[0007] In one aspect, a composition is provided comprising a compound of any embodiment disclosed herein, a pharmaceutically acceptable carrier or one or more excipients, fillers or agents (hereinafter collectively referred to as a "pharmaceutically acceptable carrier", unless otherwise stated and / or specified).

[0008] In a related aspect, provided is a medicament for treating cancer in a subject, comprising a compound of any of the embodiments disclosed herein and optionally a pharmaceutically acceptable carrier.

[0009] In a related aspect, a pharmaceutical composition is provided, comprising (i) an effective amount of a compound of any embodiment disclosed herein, wherein the effective amount of the compound is effective for treating cancer; and (ii) a pharmaceutically acceptable carrier.

[0010] In a related aspect, a pharmaceutical composition is provided comprising (i) an effective amount of a compound of any embodiment disclosed herein, wherein the compound is present in an amount effective to treat cancer when combined with a second cancer therapy; and (ii) a pharmaceutically acceptable carrier.

[0011] In other related aspects, the present technology provides methods including compounds of any aspect or embodiment disclosed herein and / or compositions of any embodiment disclosed herein and / or drugs of any embodiment disclosed herein. Such methods include methods of treating subjects suffering from cancer, wherein the method includes administering to the subject an effective amount of a compound of any embodiment disclosed herein and an effective amount of a second cancer therapy. DETAILED DESCRIPTION

[0012] As defined below, the following terms are used throughout.

[0013] As used herein and in the appended claims, unless otherwise indicated herein or clearly contradicted by context, singular articles such as "a / an" and "said" and similar references in the context of describing elements (especially in the context of the following claims) should be understood to cover both the singular and the plural. Unless otherwise indicated herein, the description of the range of values ​​herein is intended only to serve as a shorthand method for individually referring to each individual value within the range, and each individual value is incorporated into this specification as if individually described herein. Unless otherwise indicated herein or clearly contradicted by context, all methods described herein can be performed in any suitable order. Unless otherwise indicated, the use of any and all examples or exemplary language (e.g., "such as") provided herein is intended only to better illustrate the embodiments, and does not constitute a limitation on the scope of the claims. The language in this specification should not be interpreted as indicating that any unclaimed element is essential.

[0014] As used herein, "about" will be understood by one of ordinary skill in the art and will vary to some extent depending on the context in which it is used. If there is a use of the term that is unclear to one of ordinary skill in the art, "about" will mean up to plus or minus 10% of the particular term, taking into account the context in which the term is used, e.g., "about 10% by weight" will be understood to mean "9% to 11% by weight". It should be understood that when "about" precedes a term, the term should be interpreted to disclose "about" the term as well as the term not modified by "about", e.g., "about 10% by weight" discloses "9% to 11% by weight", as well as discloses "10% by weight".

[0015] The phrase "and / or" used in this disclosure should be understood to mean any one of the listed members individually or in combination of any two or more thereof, for example, "A, B and / or C" will mean "A, B, C, A and B, A and C, B and C, or a combination of A, B and C."

[0016] Generally speaking, reference to an element such as hydrogen or H is meant to include all isotopes of that element. For example, if an R group is defined to include hydrogen or H, it also includes deuterium and tritium. Thus, reference to an element such as tritium, C 14 , P 32 and S 35 The compounds of the present technology are within the scope of the present technology. Based on the disclosure herein, the procedures for inserting such tags into the compounds of the present technology will be obvious to those skilled in the art.

[0017] Typically, "substituted" refers to an organic group (e.g., an alkyl group) as defined below, wherein one or more bonds to the hydrogen atoms contained therein are replaced by bonds to non-hydrogen or non-carbon atoms. Substituted groups also include groups in which one or more bonds bound to carbon atoms or hydrogen atoms are replaced by one or more bonds (including double bonds or triple bonds) bound to heteroatoms. Therefore, unless otherwise specified, substituted groups are substituted by one or more substituents. In some embodiments, substituted groups are substituted by 1, 2, 3, 4, 5 or 6 substituents. Examples of substituents include: halogen (i.e., F, Cl, Br and I); hydroxyl; alkoxy, alkenyloxy, aryloxy, aralkyloxy, heterocyclyl, heterocyclylalkyl, heterocyclyloxy and heterocyclylalkoxy groups; carbonyl (oxo); carboxylate; ester; polyurethane; oxime; hydroxylamine; alkoxyamine; aralkyloxyamine; thiol; sulfide; sulfoxide; sulfone; sulfonyl; pentafluorosulfanyl (i.e., SF 5 ), sulfonamides; amines; N-oxides; hydrazines; hydrazides; hydrazones; azides; amides; ureas; amidines; guanidines; enamines; imides; isocyanates; isothiocyanates; cyanates; thiocyanates; imines; nitro groups; and nitriles (i.e., CN).

[0018] Substituted ring groups, such as substituted cycloalkyl, aryl, heterocyclyl and heteroaryl, also include rings and ring systems in which bonds to hydrogen atoms are replaced by bonds to carbon atoms. Thus, substituted cycloalkyl, aryl, heterocyclyl and heteroaryl may also be substituted with substituted or unsubstituted alkyl, alkenyl and alkynyl groups as defined below.

[0019] Alkyl groups include straight and branched alkyl groups with 1 to 12 carbon atoms and typically 1 to 10 carbons or in some embodiments 1 to 8, 1 to 6 or 1 to 4 carbon atoms. The alkyl group can be substituted or unsubstituted. Examples of straight chain alkyl groups include groups such as methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl and n-octyl. Examples of branched alkyl groups include but are not limited to isopropyl, isobutyl, sec-butyl, tert-butyl, neopentyl, isopentyl and 2,2-dimethylpropyl groups. Representative substituted alkyl groups can be substituted once or multiple times by substituents such as substituents listed above, and include but are not limited to haloalkyl (e.g., trifluoromethyl), hydroxyalkyl, sulfanyl, aminoalkyl, alkylaminoalkyl, dialkylaminoalkyl, alkoxyalkyl, carboxyalkyl, etc.

[0020] Cycloalkyl groups include monocyclic alkyl, bicyclic alkyl or tricyclic alkyl groups having 3 to 12 carbon atoms (or in some embodiments, 3 to 10, 3 to 8 or 3 to 4, 5 or 6 carbon atoms) in the ring. Cycloalkyl groups can be substituted or unsubstituted. Exemplary monocyclic cycloalkyl groups include but are not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl. In some embodiments, the cycloalkyl group has 3 to 8 ring members, while in other embodiments, the number of ring carbon atoms is in the range of 3 to 5, 3 to 6 or 3 to 7. Bicyclic ring systems and tricyclic ring systems include both bridged cycloalkyl groups and condensed rings, such as but not limited to bicyclic [2.1.1] hexane, adamantyl, decahydronaphthyl, etc. Substituted cycloalkyl groups can be substituted once or multiple times by non-hydrogen and non-carbon groups as defined above. However, substituted cycloalkyl groups also include rings substituted by straight or branched alkyl as defined above. Representative substituted cycloalkyl groups may be monosubstituted or substituted more than once, such as, but not limited to, 2,2-, 2,3-, 2,4-, 2,5-, or 2,6-disubstituted cyclohexyl groups, which may be substituted with substituents such as those listed above.

[0021] Cycloalkylalkyl group is an alkyl group as defined above, wherein the hydrogen bond or carbon bond of the alkyl group is replaced by a key to the cycloalkyl group such as defined above.Cycloalkylalkyl can be substituted or unsubstituted.In certain embodiments, cycloalkylalkyl has 4 to 16 carbon atoms, 4 to 12 carbon atoms and typically 4 to 10 carbon atoms.Substituted cycloalkylalkyl can be substituted at both the alkyl, cycloalkyl or alkyl and cycloalkyl parts of the group.Representative substituted cycloalkylalkyl group can be monosubstituted or substituted more than once, such as but not limited to being monosubstituted, disubstituted or trisubstituted by substituents such as substituents listed above.

[0022] Alkenyl groups include straight and branched alkyl groups as defined above, except that there is at least one double bond between two carbon atoms. Alkenyl groups can be substituted or unsubstituted. Alkenyl groups have 2 to 12 carbon atoms and typically 2 to 10 carbons or in some embodiments 2 to 8, 2 to 6, or 2 to 4 carbon atoms. In some embodiments, alkenyl groups have one, two, or three carbon-carbon double bonds. Examples include, but are not limited to, vinyl, allyl, -CH=CH(CH 3 ), -CH=C(CH 3 ) 2 、-C(CH 3 )=CH 2 、-C(CH 3 )=CH(CH 3 )、-C(CH 2 CH 3 )=CH 2 etc. Representative substituted alkenyl groups can be mono-substituted or substituted more than once, such as, but not limited to, mono-, di-, or tri-substituted with substituents such as those listed above.

[0023] Cycloalkenyl groups include cycloalkyl as defined above with at least one double bond between two carbon atoms.Cycloalkenyl groups can be substituted or unsubstituted.In certain embodiments, cycloalkenyl groups can have one, two or three double bonds, but do not include aromatic compounds.Cycloalkenyl groups have 4 to 14 carbon atoms, or in certain embodiments, 5 to 14 carbon atoms, 5 to 10 carbon atoms or even 5,6,7 or 8 carbon atoms.The example of cycloalkenyl groups includes cyclohexenyl, cyclopentenyl, cyclohexadienyl, cyclobutadienyl and cyclopentadienyl.

[0024] Cycloalkenylalkyl group is an alkyl group as defined above, wherein the hydrogen bond or carbon bond of the alkyl group is replaced by the key to the cycloalkenyl group as defined above.Cycloalkenylalkyl group can be substituted or unsubstituted.Substituted cycloalkenylalkyl group can be substituted at the alkyl part of the group, at the cycloalkenyl part or at both the alkyl part and the cycloalkenyl part.Representational substituted cycloalkenylalkyl group can be substituted once or many times with a substituent such as the substituent listed above.

[0025] Alkynyl groups include straight and branched alkyl groups as defined above, except that there is at least one triple bond between two carbon atoms. Alkynyl groups can be substituted or unsubstituted. Alkynyl groups have 2 to 12 carbon atoms and typically 2 to 10 carbons or in some embodiments 2 to 8, 2 to 6 or 2 to 4 carbon atoms. In some embodiments, alkynyl groups have one, two or three carbon-carbon triple bonds. Examples include, but are not limited to -C≡CH, -C≡CCH3 、-CH 2 C≡CCH 3 and -C≡CCH 2 CH(CH 2 CH 3 ) 2 etc. Representative substituted alkynyl groups can be mono-substituted or substituted more than once, such as, but not limited to, mono-, di-, or tri-substituted with substituents such as those listed above.

[0026] Aryl groups are cyclic aromatic hydrocarbons that do not contain heteroatoms. Aryl groups herein include monocyclic, bicyclic and tricyclic ring systems. Aryl groups can be substituted or unsubstituted. Therefore, aryl groups include but are not limited to phenyl, azulenyl, heptenyl, biphenyl, fluorenyl, phenanthrenyl, anthracenyl, indenyl, indanyl, pentalenyl and naphthyl groups. In some embodiments, the aryl group contains 6 to 14 carbons in the ring portion of the group, and in other embodiments contains 6 to 12 or even 6 to 10 carbon atoms. In some embodiments, the aryl group is phenyl or naphthyl. The phrase "aryl group" includes groups containing fused rings, such as fused aromatic-aliphatic ring systems (e.g., indanyl, tetrahydronaphthyl, etc.). Representative substituted aryl groups can be monosubstituted (e.g., tolyl) or substituted more than once. For example, monosubstituted aryl groups include, but are not limited to, 2-, 3-, 4-, 5-, or 6-substituted phenyl or naphthyl, which may be substituted with substituents such as those listed above.

[0027] Aralkyl group is an alkyl group as defined above, wherein the hydrogen bond or carbon bond of the alkyl group is replaced by a key to an aryl group as defined above. Aralkyl group can be substituted or unsubstituted. In certain embodiments, the aralkyl group contains 7 to 16 carbon atoms, 7 to 14 carbon atoms or 7 to 10 carbon atoms. Substituted aralkyl group can be substituted at both the alkyl, aryl or alkyl and aryl parts of the group. Representative aralkyl groups include but are not limited to benzyl and phenethyl groups and fused (cycloalkylaryl) alkyl groups such as 4-indanylethyl. Representative substituted aralkyl groups can be substituted once or multiple times with substituents (such as the substituents listed above).

[0028] Heterocyclyl groups include aromatic (also referred to as heteroaryl) and non-aromatic ring compounds containing 3 or more ring members, one or more of which are heteroatoms, such as, but not limited to, N, O, and S. Heterocyclyl groups may be substituted or unsubstituted. In some embodiments, heterocyclyl groups contain 1, 2, 3, or 4 heteroatoms. In some embodiments, heterocyclyl groups include monocyclic, bicyclic, and tricyclic rings having 3 to 16 ring members, while other such groups have 3 to 6, 3 to 10, 3 to 12, or 3 to 14 ring members. Heterocyclyl groups include aromatic, partially unsaturated, and saturated ring systems, such as, for example, imidazolyl, imidazolinyl, and imidazolidinyl groups. The phrase "heterocyclyl group" includes fused ring species, including those containing fused aromatic and non-aromatic groups, such as, for example, benzotriazolyl, 2,3-dihydrobenzo[1,4]dioxinyl, and benzo[1,3]dioxolyl. The phrase also includes bridged polycyclic ring systems containing heteroatoms, such as, but not limited to, quinuclidine. The phrase includes heterocyclyl groups having other groups (such as alkyl, oxo or halo groups) bonded to one of the ring members, referred to as "substituted heterocyclyl groups". Heterocyclyl groups include, but are not limited to, aziridinyl, azetidinyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, thiazolidinyl, tetrahydrothienyl, oxolanyl, dioxolyl, furanyl, thienyl, pyrrolyl, pyrrolinyl, imidazolyl, imidazolinyl, pyrazolyl, pyrazolinyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, thiazolinyl, isothiazolyl, thiadiazolyl, oxadiazolyl, piperidinyl, piperazinyl , morpholinyl, thiomorpholinyl, tetrahydropyranyl, tetrahydrothiopyranyl, oxathiolane, dioxanyl, dithianyl, pyranyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, triazinyl, dihydropyridyl, dihydrodithiopyranyl, dihydrodisulfinyl, homopiperazinyl, quinuclidine, indolyl, indolinyl, isoindolyl, azaindolyl (pyrrolopyridinyl), indazolyl, indolizinyl, benzotriazolyl, benzimidazolyl , benzofuranyl, benzothienyl, benzothiazolyl, benzoxadiazolyl, benzoxazininyl, benzodithienyl, benzoxathienyl, benzothiazinyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, benzo[1,3]dioxolyl, pyrazolopyridinyl, imidazopyridinyl (azabenzimidazolyl), triazolopyridinyl, isoxazolopyridinyl, purinyl, xanthinyl, adeninyl, guaninyl, quinoline quinolinyl, isoquinolinyl, quinolizinyl, quinoxalinyl, quinazolinyl, cinnolinyl, phthalazinyl, naphthyridinyl, pteridinyl, thiaphthyl, dihydrobenzothiazinyl, dihydrobenzofuranyl, dihydroindolyl, dihydrobenzodioxinyl, tetrahydroindolyl, tetrahydroindazolyl, tetrahydrobenzimidazolyl, tetrahydrobenzotriazolyl, tetrahydropyrrolopyridinyl, tetrahydropyrazolopyridinyl, tetrahydroimidazopyridinyl, tetrahydrotriazolopyridinyl and tetrahydroquinolinyl.Representative substituted heterocyclyl groups can be monosubstituted or substituted more than once, such as, but not limited to, pyridinyl or morpholinyl groups substituted at the 2-, 3-, 4-, 5-, or 6-position or disubstituted with various substituents such as those listed above.

[0029] Heteroaryl groups are aromatic ring compounds containing 5 or more ring members, one or more of which are heteroatoms such as, but not limited to, N, O, and S. Heteroaryl groups may be substituted or unsubstituted. Heteroaryl groups include, but are not limited to, groups such as pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, thienyl, benzothienyl, furanyl, benzofuranyl, indolyl, azaindolyl (pyrrolopyridinyl), indazolyl, benzimidazolyl, imidazopyridinyl (azabenzimidazolyl), pyrazolopyridinyl, triazolopyridinyl, benzotriazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, imidazopyridinyl, isoxazolopyridinyl, thianaphthyl, purinyl, xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, quinoxalinyl, and quinazolinyl. Heteroaryl groups include fused ring compounds in which all rings are aromatic, such as indolyl groups, and include fused ring compounds in which only one of the rings is aromatic, such as 2,3-dihydroindolinyl groups. Representative substituted heteroaryl groups can be substituted one or more times with various substituents such as those listed above.

[0030] Heterocyclylalkyl group is an alkyl group as defined above, wherein the hydrogen bond or carbon bond of the alkyl group is replaced by a bond to a heterocyclyl group as defined above. Heterocyclylalkyl group can be substituted or unsubstituted. Substituted heterocyclylalkyl group can be substituted at the alkyl portion of the group, at the heterocyclyl portion, or at both the alkyl portion and the heterocyclyl portion. Representative heterocyclylalkyl groups include but are not limited to morpholine-4-yl-ethyl, furan-2-yl-methyl, imidazole-4-yl-methyl, pyridine-3-yl-methyl, oxacyclopentane-2-yl-ethyl and indol-2-yl-propyl. Representative substituted heterocyclylalkyl groups can be substituted once or multiple times with substituents (such as the substituents listed above).

[0031] Heteroaralkyl groups are alkyl groups as defined above, wherein the hydrogen bonds or carbon bonds of the alkyl groups are replaced by the keys to heteroaryl groups as defined above.Heteroaralkyl groups can be substituted or unsubstituted.Substituted cycloalkenylalkyl groups can be substituted at the alkyl portion of the group, at the heteroaryl portion, or at both the alkyl portion and the heteroaryl portion.Representative substituted heteroaralkyl groups can be substituted once or many times with substituents (such as the substituents listed above).

[0032] Groups described herein that have two or more points of attachment (i.e., divalent, trivalent, or multivalent) within the compounds of the present technology are named by using the suffix "ene." For example, a divalent alkyl group is an alkylene group, a divalent aryl group is an arylene group, a divalent heteroaryl group is a divalent heteroarylene group, etc. Substituted groups that have a single point of attachment to the compounds of the present technology are not referred to using the "ene" designation. Thus, for example, chloroethyl is not referred to herein as vinyl chloride.

[0033] Alkoxy groups are hydroxyl groups (-OH) in which the bond to the hydrogen atom is replaced by the bond to the carbon atom of the substituted or unsubstituted alkyl group as defined above. Alkoxy groups can be substituted or unsubstituted. Examples of straight chain alkoxy groups include but are not limited to methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy, etc. Examples of branched chain alkoxy groups include but are not limited to isopropoxy, sec-butoxy, tert-butoxy, isopentyloxy, isohexyloxy, etc. Examples of cycloalkoxy groups include but are not limited to cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, etc. Representative substituted alkoxy groups can be substituted once or multiple times by substituents such as those listed above.

[0034] As used herein, the terms "alkanoyl" and "alkanoyloxy" may refer to -C(O)-alkyl and -OC(O)-alkyl groups, respectively, each containing 2 to 5 carbon atoms. Similarly, "aroyl" and "aroyloxy" refer to -C(O)-aryl and -OC(O)-aryl groups.

[0035] The terms "aryloxy" and "arylalkoxy" refer to a substituted or unsubstituted aryl group bonded to an oxygen atom and a substituted or unsubstituted aralkyl group bonded to an oxygen atom at the alkyl group, respectively. Examples include, but are not limited to, phenoxy, naphthoxy, and benzyloxy. Representative substituted aryloxy and arylalkoxy groups may be substituted one or more times with substituents such as those listed above.

[0036] The term "carboxylate" as used herein refers to a -COOH group.

[0037] As used herein, the term "ester" refers to -COOR 70 and –C(O)OG groups. 70R is a substituted or unsubstituted alkyl, cycloalkyl, alkenyl, alkynyl, aryl, aralkyl, heterocyclylalkyl or heterocyclyl group as defined herein. G is a carboxylate protecting group. Carboxylate protecting groups are well known to those of ordinary skill in the art. An extensive list of protecting groups for carboxylate groups is found in Protective Groups in Organic Synthesis, Greene, TW; Wuts, PGM, John Wiley & Sons, New York, NY, (3rd edition, 1999), and the procedures set forth in the document can be used to add or remove protecting groups, and the document is hereby incorporated by reference in its entirety and for any and all purposes, as if fully set forth herein.

[0038] The term "amide" (or "amido") includes C-amide and N-amide groups, i.e., -C(O)NR 71 R 72 Group and –NR 71 C(O)R 72 Group. R 71 and R 72 is independently hydrogen, or a substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heterocyclylalkyl or heterocyclyl group as defined herein. Thus, amido groups include but are not limited to carbamoyl (-C(O)NH 2 ) and carboxamide groups (-NHC(O)H). In some embodiments, the amide is -NR 71 C(O)-(C 1-5 In some embodiments, the amide is -NHC(O)-alkyl and the group is referred to as a "carbonylamino" group, and in other embodiments, the amide is -NHC(O)-alkyl and the group is referred to as a "alkanoylamino" group.

[0039] As used herein, the term "nitrile" or "cyano" refers to a -CN group.

[0040] The carbamate groups include N-carbamate groups and O-carbamate groups, namely -NR 73 C(O)OR 74 Group and -OC(O)NR 73 R 74 Group. R 73 and R 74 R is independently a substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heterocyclylalkyl or heterocyclyl group as defined herein. 73 It can also be H.

[0041] As used herein, the term "amine" (or "amino") refers to a -NR 75 R76 Group, where R 75 and R 76 is independently hydrogen, or a substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heterocyclylalkyl, or heterocyclyl group as defined herein. In some embodiments, the amine is an alkylamino, dialkylamino, arylamino, or alkylarylamino. In other embodiments, the amine is NH 2 , methylamino, dimethylamino, ethylamino, diethylamino, propylamino, isopropylamino, phenylamino or benzylamino.

[0042] The term "sulfonamide" includes S- and N-sulfonamide groups, i.e., -SO 2 NR 78 R 79 Group and –NR 78 SO 2 R 79 Group. R 78 and R 79 are independently hydrogen, or a substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heterocyclylalkyl or heterocyclyl group as defined herein. Thus, sulfonamido groups include, but are not limited to, sulfamoyl (-SO 2 NH 2 In some embodiments herein, the sulfonamide group is -NHSO 2 -alkyl and is known as an "alkylsulfonylamino" group.

[0043] The term "thiol" refers to a -SH group, while "sulfide" includes -SR 80 The group, “sulfoxide” includes –S(O)R 81 The group "sulfone" includes -SO 2 R 82 group, and "sulfonyl" includes –SO 2 OR 83 . R 80 , R 81 , R 82 and R 83 Each is independently a substituted or unsubstituted alkyl, cycloalkyl, alkenyl, alkynyl, arylarylalkyl, heterocyclyl or heterocyclylalkyl group as defined herein. In some embodiments, the sulfide is an alkylthio group, -S-alkyl.

[0044] The term "urea" refers to -NR 84 -C(O)-NR 85 R 86 Group. R 84 , R 85 and R 86Groups are independently hydrogen, or a substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heterocyclyl or heterocyclylalkyl group as defined herein.

[0045] The term "amidine" refers to -C(NR 87 )NR 88 R 89 and –NR 87 C(NR 88 )R 89 , where R 87 , R 88 and R 89 Each is independently hydrogen, or a substituted or unsubstituted alkyl, cycloalkyl, alkenyl, alkynyl, arylaralkyl, heterocyclyl or heterocyclylalkyl group as defined herein.

[0046] The term "guanidine" refers to -NR 90 C(NR 91 )NR 92 R 93 , where R 90 , R 91 , R 92 and R 93 Each is independently hydrogen, or a substituted or unsubstituted alkyl, cycloalkyl, alkenyl, alkynyl, arylaralkyl, heterocyclyl or heterocyclylalkyl group as defined herein.

[0047] The term "enamine" refers to a -C(R 94 )=C(R 95 )NR 96 R 97 and –NR 94 C(R 95 )=C(R 96 )R 97 , where R 94 , R 95 , R 96 and R 97 Each is independently hydrogen, a substituted or unsubstituted alkyl, cycloalkyl, alkenyl, alkynyl, arylaralkyl, heterocyclyl or heterocyclylalkyl group as defined herein.

[0048] As used herein, the term "halogen" or "halo" refers to bromine, chlorine, fluorine or iodine. In some embodiments, the halogen is fluorine. In other embodiments, the halogen is chlorine or bromine.

[0049] As used herein, the term "hydroxyl" may refer to -OH or its ionized form -OH. – A "hydroxyalkyl" group is an alkyl group substituted with a hydroxy group, such as HO-CH 2 -.

[0050] The term "imide" refers to -C(O)NR 98 C(O)R 99 , where R 98 and R 99 Each is independently hydrogen, or a substituted or unsubstituted alkyl, cycloalkyl, alkenyl, alkynyl, arylaralkyl, heterocyclyl or heterocyclylalkyl group as defined herein.

[0051] The term "imine" refers to –CR 100 (NR 101 ) and –N(CR 100 R 101 ) group, wherein R 100 and R 101 are each independently hydrogen or a substituted or unsubstituted alkyl, cycloalkyl, alkenyl, alkynyl, arylarylalkyl, heterocyclyl or heterocyclylalkyl group as defined herein, provided that R 100 and R 101 Both are not hydrogen at the same time.

[0052] As used herein, the term "nitro" refers to -NO 2 Group.

[0053] As used herein, the term "trifluoromethyl" refers to -CF 3 .

[0054] As used herein, the term "trifluoromethoxy" refers to -OCF 3 .

[0055] The term "azido" refers to -N 3 .

[0056] The term "trialkylammonium" refers to -N(alkyl) 3 The trialkylammonium group is positively charged and therefore typically has an associated anion, such as a halide anion.

[0057] The term "isocyano" refers to -NC.

[0058] The term "isothiocyanato" refers to -NCS.

[0059] The term "pentafluorosulfanyl" refers to -SF 5 .

[0060] As will be understood by those skilled in the art, for any and all purposes, particularly in providing written description, all scopes disclosed herein also encompass any and all possible sub-ranges and combinations of sub-ranges thereof.Any listed scope can be easily considered as fully describing and enabling the same scope to be decomposed into at least equal half, one-third, one-quarter, one-fifth, one-tenth, etc. As a non-limiting example, each scope discussed herein can be easily decomposed into lower third, middle third, and upper third, etc. As will be understood by those skilled in the art, all languages ​​such as "at most", "at least", "greater than", "less than", etc. include the listed numbers and refer to the scope of the sub-ranges discussed above subsequently. Finally, as will be understood by those skilled in the art, scope includes each individual member.Therefore, for example, a group with 1 to 3 atoms refers to a group with 1, 2 or 3 atoms.Similarly, a group with 1-5 atoms refers to a group with 1, 2, 3, 4 or 5 atoms, etc.

[0061] As understood by one of ordinary skill in the art, "molecular weight" (also known as "relative molar mass") is a dimensionless quantity but is converted to molar mass by multiplying by 1 gram / mole or by 1 Da, e.g., a compound with a weight average molecular weight of 5,000 has a weight average molar mass of 5,000 g / mol and a weight average molar mass of 5,000 Da.

[0062] Pharmaceutically acceptable salts of the compounds described herein are within the scope of the present technology and include acid addition salts or base addition salts that retain the desired pharmacological activity and are not biologically undesirable (e.g., the salts do not have undue toxicity, allergenicity or irritation, and are bioavailable). When the compounds of the present technology have a basic group (such as, for example, an amino group), pharmaceutically acceptable salts can be formed by inorganic acids (such as hydrochloric acid, hydroboric acid, nitric acid, sulfuric acid, and phosphoric acid), organic acids (such as alginic acid, formic acid, acetic acid, benzoic acid, gluconic acid, fumaric acid, oxalic acid, tartaric acid, lactic acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, benzenesulfonic acid, naphthalenesulfonic acid, and p-toluenesulfonic acid) or acidic amino acids (such as aspartic acid and glutamic acid). When the compounds of the present technology have an acidic group (such as, for example, a carboxylic acid group), it can react with metals such as alkali metals and alkaline earth metals (e.g., Na + , Li + , K + , Ca 2+ Mg 2+ 、Zn 2+), ammonia or organic amines (e.g., dicyclohexylamine, trimethylamine, triethylamine, pyridine, picoline, ethanolamine, diethanolamine, triethanolamine) or basic amino acids (e.g., arginine, lysine and ornithine) to form salts. Such salts can be prepared in situ during the isolation and purification of the compound, or by reacting the purified compound in free base or free acid form with a suitable acid or base, respectively, and isolating the salt thus formed.

[0063] Those skilled in the art will understand that the compounds of the present technology can exhibit tautomerism, conformational isomerism, geometric isomerism and / or stereoisomerism. Since the chemical formula drawings in the specification and claims can only represent one possible tautomerism, conformational isomerism, stereochemical or geometric isomeric form, it should be understood that the present technology covers any tautomerism, conformational isomerism, stereochemical and / or geometric isomeric form of the compound having one or more of the utilities described herein, as well as mixtures of these various forms.

[0064] "Tautomers" refer to isomeric forms of compounds that are in equilibrium with each other. The presence and concentration of isomeric forms will depend on the environment in which the compound exists and may differ depending on, for example, whether the compound is solid or in an organic solution or aqueous solution. For example, in aqueous solution, quinolones may exhibit the following isomeric forms (which are also referred to as tautomers of each other):

[0065]

[0066] As another example, guanidine can exhibit the following isomeric forms (also referred to as tautomers of one another) in protic organic solutions:

[0067]

[0068] As representative compounds are limited by structural formulae, it is understood that all formulae of compounds described herein represent all tautomeric forms of the compounds and are within the scope of the present technology.

[0069] Unless a specific stereochemistry is explicitly indicated, stereoisomers (also referred to as optical isomers) of a compound include all chiral, diastereomeric, and racemic forms of the structure. Thus, the compounds used in the present technology include enriched or resolved optical isomers at any or all asymmetric atoms, as apparent from the description. Both racemic and diastereomeric mixtures, as well as individual optical isomers, can be separated or synthesized to be substantially free of their enantiomeric or diastereomeric partners, and these stereoisomers are within the scope of the present technology.

[0070] The compounds of the present technology may exist as solvates (especially hydrates). Hydrates may be formed during the manufacture of the compound or a composition comprising the compound, or hydrates may be formed over time due to the hygroscopic properties of the compound. The compounds of the present technology may also exist as organic solvates (including DMF, ether and alcohol solvates, etc.). The identification and preparation of any particular solvate is within the skill of a person of ordinary skill in synthetic organic chemistry or medicinal chemistry.

[0071] Throughout this disclosure, various publications, patents, and published patent specifications are referenced by identifying citations. Arabic numerals referring to the references are also included in this disclosure, and their complete bibliographic details are provided after the Examples section. The disclosures of these publications, patents, and published patent specifications are hereby incorporated by reference into this disclosure to more fully describe the present technology.

[0072] This technology

[0073] Poly ADP ribose polymerase 1 (PARP1) is a member of a family of proteins involved in numerous cellular processes such as DNA repair, genome stability, and programmed cell death. PARP1 is an ADP-ribosyltransferase that uses NAD+ as a substrate to modify proteins, including itself, through PARylation and detects single-strand DNA breaks, thereby playing an important role in homologous recombination and DNA repair by signaling the enzymatic processes involved in repairing single-strand DNA breaks.

[0074] Several PARP1 inhibitors have been approved for the treatment of breast and ovarian cancers deficient in other DNA repair mechanisms, and additional PARP inhibitors are currently in clinical trials for the treatment of ovarian cancer, pancreatic and biliary malignancies, glioblastoma, lung cancer, and prostate cancer. BRCA1 / 2-deficient cancers are extremely sensitive to PARP1 inhibition, with PARP inhibition being lethal for cells with loss-of-function mutations in BRCA1 and BRCA2. Reduced homologous recombination (HR) capacity through other mechanisms also sensitizes cells to PARP inhibitors. Cells lacking DNA repair defects are typically 1000-fold less susceptible to PARP inhibitors than cells with such defects. PARP inhibitors reduce PARylation and trap PARP1 on DNA, leading to replication fork collapse and cell death.

[0075] However, current PARP1 inhibitors such as talazoparib, olaparib, rucaparib and niraparib do not have good central nervous system (CNS) penetration, and veliparib is ineffective for most tumors. See, for example, Gupta, Shiv K. et al. "PARP inhibitors for sensitization of alkylation chemotherapy inglioblastoma: impact of blood-brain barrier and molecular heterogeneity" Frontiers in oncology 8 (2019): 670; Kizilbash, SH et al. "Restricted delivery of talazoparib across the blood-brain barrier limits the sensitizing effects of PARP inhibition on Temozolomide therapy in glioblastoma" Mol. Cancer Ther, 16 (2017): 2735-2746.

[0076] Therefore, there is a need for PARP inhibitors with improved central nervous system penetration for the treatment of brain cancers such as glioblastoma, neuroblastoma, and medulloblastoma.

[0077] The present technology responds to these needs and provides additional advantages. According to internal proprietary prediction models, it is expected that the compounds of the present technology show desirable blood-brain barrier (BBB) ​​permeability. These proprietary prediction models are further verified by in vitro assays, which have been fully confirmed to predict the BBB permeability of living subjects, and are further verified by in vivo experiments. Therefore, the present technology provides compounds with advantageously improved central nervous system penetration, and compositions and methods particularly suitable for treating central nervous system cancer.

[0078] Therefore, in one aspect, the present technology provides a compound according to formula I

[0079]

[0080] or a pharmaceutically acceptable salt and / or solvate thereof, wherein X 1 is H, F or Cl; X 2 Is NH or NR 3 ; R 1 and R 2 One of is H, halo, alkyl, alkenyl, alkynyl, cycloalkyl or heterocyclyl, and R1 and R 2 The other one of R is an aryl group, a heteroaryl group or a non-aromatic heterocyclic group; 3 It is an alkyl, cycloalkyl, alkylene and non-aromatic heterocyclic group. For ease of reference, the compounds included in any aspect or embodiment herein may be referred to as "a compound of the present technology / compounds of the present technology" etc. anywhere in this disclosure. Similarly, for ease of reference, the compositions, drugs and pharmaceutical compositions of the present technology may be collectively referred to as "compositions", "compositions of the present technology" etc. in this article.

[0081] In any embodiment herein, the compound of formula I may be a compound of formula IA

[0082]

[0083] or a pharmaceutically acceptable salt and / or solvate thereof.

[0084] In any embodiment herein, the compound of formula I may be a compound of formula IB

[0085]

[0086] or a pharmaceutically acceptable salt and / or solvate thereof.

[0087] In any of the embodiments herein, it is possible that R 1 is aryl, heteroaryl or non-aromatic heterocyclic group; and R 2 is H, halo, alkyl, alkenyl, alkynyl, cycloalkyl or non-aromatic heterocyclyl. In any embodiment herein, it is possible that R 1 is aryl, heteroaryl or non-aromatic heterocyclic group; and R 2 is H, alkyl, cycloalkyl or non-aromatic heterocyclyl. In any embodiment herein, it is possible that X 1 is F. In any embodiment herein, it is possible that X 2 It's NH.

[0088] In one aspect, a composition is provided, comprising a compound of any embodiment disclosed herein, a pharmaceutically acceptable carrier or one or more excipients, fillers or agents (hereinafter collectively referred to as "pharmaceutically acceptable carriers", unless otherwise stated and / or specified). In a related aspect, a drug for treating cancer in a subject is provided, comprising a compound of any embodiment disclosed herein and optionally a pharmaceutically acceptable carrier. The drug of any embodiment herein may include an effective amount of a compound for treating cancer when combined with a second cancer therapy (such as radiotherapy, a monoclonal antibody and / or a chemotherapeutic agent). In any embodiment herein, the cancer may be breast cancer, ovarian cancer, pancreatic cancer, biliary tract cancer, lung cancer, prostate cancer and / or CNS cancer (such as brain cancer). CNS cancer may be glioblastoma, neuroblastoma and / or medulloblastoma. In a related aspect, a pharmaceutical composition is provided, comprising (i) an effective amount of a compound of any embodiment disclosed herein, wherein the effective amount of the compound is effective for treating cancer; and (ii) a pharmaceutically acceptable carrier. In any embodiment herein, the cancer may be breast cancer, ovarian cancer, pancreatic cancer, biliary tract cancer, lung cancer, prostate cancer and / or CNS cancer (such as brain cancer). In any embodiment herein, cancer can be breast cancer, ovarian cancer, pancreatic cancer, biliary tract cancer, lung cancer, prostate cancer and / or CNS cancer (such as brain cancer). In related aspects, a pharmaceutical composition is provided, which comprises (i) an effective amount of a compound of any embodiment disclosed herein, wherein the compound is present in an amount effective for treating cancer when combined with a second cancer therapy (such as radiotherapy, a monoclonal antibody and / or a chemotherapeutic agent); and (ii) a pharmaceutically acceptable carrier. In any embodiment herein, cancer can be breast cancer, ovarian cancer, pancreatic cancer, biliary tract cancer, lung cancer, prostate cancer and / or CNS cancer (such as brain cancer). In other related aspects, the present technology provides a method for including a compound of any aspect or embodiment disclosed herein and / or a composition of any embodiment disclosed herein and / or a drug of any embodiment disclosed herein.

[0089] "Effective amount" refers to the amount of the compound or composition required to produce the desired effect. An example of an effective amount includes an amount or dosage that produces an acceptable level of toxicity and bioavailability for treatment (drug) purposes, including but not limited to reducing tumor mass. In any aspect or embodiment of the composition, pharmaceutical composition and method disclosed herein including the compound of the present technology (collectively referred to herein as "any embodiment of the present invention", "any embodiment disclosed herein", etc.), the effective amount can be an amount that can be effective for treating cancer (such as breast cancer, ovarian cancer, pancreatic cancer, biliary tract cancer, lung cancer, prostate cancer and / or CNS cancer), treating tumors and / or shrinking tumors. For example, the effective amount of any embodiment of the present invention including the compound of the present technology can be about 0.01 μg to about 200 mg of the compound (such as about 0.1 μg to about 50 mg of the compound or about 10 μg to about 20 mg of the compound). The method and use according to the present technology may include an effective amount of the compound of any embodiment disclosed herein. In any aspect or embodiment disclosed herein, the effective amount can be determined relative to the subject. As used herein, "subject" or "patient" is a mammal, such as a cat, dog, rodent or primate. Typically, the subject is a human, and preferably a human suffering from or suspected of suffering from pain.The terms "subject" and "patient" are used interchangeably.

[0090] Therefore, the present technology provides pharmaceutical compositions and drugs, which include compounds of any embodiment disclosed herein (or compositions of any embodiment disclosed herein) and a pharmaceutically acceptable carrier. The composition can be used in the methods and treatments described herein. The pharmaceutical composition can be packaged in unit dosage form. The unit dosage form can effectively treat cancer (such as breast cancer, ovarian cancer, pancreatic cancer, biliary tract cancer, lung cancer, prostate cancer and / or CNS cancer). When administered to a subject in need thereof, the unit dosage form can be effective for treating tumors by reducing tumor volume. In general, the unit dose of the compound containing the present technology will vary according to patient considerations. These considerations include, for example, age, regimen, condition, sex, extent of disease, contraindications, concomitant therapy, and the like. Exemplary unit doses based on these considerations can also be adjusted or modified by a physician skilled in the art. For example, the unit dose for a patient containing a compound of the present technology can be in the range of 1×10 -4 g / kg to 1 g / kg, preferably 1×10 –3The dosage of the compound of the present technology may also vary between 0.01 mg / kg and 100 mg / kg, or preferably between 0.1 mg / kg and 10 mg / kg. Suitable unit dosage forms include, but are not limited to, parenteral solutions, oral solutions, powders, tablets, pills, soft capsules, capsules, lozenges, suppositories, patches, nasal sprays, injections, implantable sustained-release preparations, mucoadhesive films, local protective lacquers, lipid complexes, liquids, etc.

[0091] Pharmaceutical compositions and drugs can be prepared by mixing one or more compounds and / or compositions of the present technology with pharmaceutically acceptable carriers, excipients, adhesives, diluents, etc. Such compositions can be in the form of, for example, granules, powders, tablets, capsules, syrups, suppositories, injections, emulsions, elixirs, suspensions, or solutions. The compositions of the present invention can be formulated for a variety of routes of administration, such as oral, parenteral, topical, rectal, nasal, vaginal administration, or administration via an implantable reservoir. Parenteral or systemic administration includes, but is not limited to, subcutaneous, intravenous, intraperitoneal, and intramuscular injections. The following dosage forms are given by way of example and should not be construed as limiting the existing present technology.

[0092] For oral, buccal and sublingual administration, powders, suspensions, granules, tablets, pills, capsules, soft capsules and caplets are acceptable as solid dosage forms. These can be (for example) prepared by mixing one or more existing compounds of the present technology or their pharmaceutically acceptable salts or tautomers with at least one additive (such as starch or other additives). Suitable additives are sucrose, lactose, cellulose sugar, mannitol, maltitol, dextran, starch, agar, alginate, chitin, chitosan, pectin, tragacanth, gum arabic, gelatin, collagen, casein, albumin, synthetic or semi-synthetic polymers or glycerides. Optionally, oral dosage forms can contain other ingredients to help use, such as inactive diluents or lubricants (such as magnesium stearate) or preservatives (such as parabens or sorbic acid) or antioxidants (such as ascorbic acid, tocopherol or cysteine), disintegrants, adhesives, thickeners, buffers, sweeteners, flavoring agents or aromatics. Tablets and pills can be further processed with suitable coating materials known in the art.

[0093] Liquid dosage forms for oral administration can be in the form of pharmaceutically acceptable emulsions, syrups, elixirs, suspensions and solutions, which may contain inert diluents (such as water). Pharmaceutical preparations and medicaments can be prepared as liquid suspensions or solutions using sterile liquids such as, but not limited to, oils, water, alcohols and combinations of these. For oral or parenteral administration, pharmaceutically suitable surfactants, suspending agents, emulsifiers may be added.

[0094] As noted above, the suspension may include oil. Such oils include, but are not limited to, peanut oil, sesame oil, cottonseed oil, corn oil, and olive oil. Suspension formulations may also contain esters of fatty acids, such as ethyl oleate, isopropyl myristate, fatty acid glycerides, and acetylated fatty acid glycerides. Suspension formulations may include alcohols, such as, but not limited to, ethanol, isopropanol, hexadecanol, glycerol, and propylene glycol. Ethers may also be used in suspension formulations, such as, but not limited to, poly(ethylene glycol), petroleum hydrocarbons such as mineral oil and petrolatum; and water.

[0095] Injectable dosage forms generally include aqueous suspensions or oily suspensions, which can be prepared using suitable dispersants or wetting agents and suspending agents. Injectable forms can be in solution phase or in suspension form (which is prepared with a solvent or diluent). Acceptable solvents or vehicles include sterile water, Ringer's solution or isotonic saline solution. Alternatively, sterile oil can be used as a solvent or suspending agent. Typically, oils or fatty acids are non-volatile, including natural or synthetic oils, fatty acids, monoglycerides, diglycerides or triglycerides.

[0096] For injection, the pharmaceutical preparation and / or drug can be a powder suitable for reconstitution with an appropriate solution as described above. Examples of these include, but are not limited to, freeze-dried, rotary-dried or spray-dried powders, amorphous powders, granules, precipitates or particles. For injection, the preparation can optionally contain a stabilizer, a pH regulator, a surfactant, a bioavailability regulator and combinations of these.

[0097] The compounds of the present technology can be administered to the lungs by inhalation through the nose or mouth. Suitable pharmaceutical formulations for inhalation include solutions, sprays, dry powders or aerosols, which contain any appropriate solvents and optional other compounds, such as but not limited to stabilizers, antimicrobial agents, antioxidants, pH regulators, surfactants, bioavailability regulators and combinations of these. Carriers and stabilizers vary with the requirements of specific compounds, but generally include nonionic surfactants (Tween, Pluronic or polyethylene glycol), harmless proteins such as serum albumin, dehydrated sorbitan esters, oleic acid, lecithin, amino acids (such as glycine), buffers, salts, sugars and / or sugar alcohols. Aqueous and non-aqueous (e.g., in fluorocarbon propellants) aerosols are commonly used to deliver compounds of the present technology by inhalation.

[0098] The dosage forms for topical (including buccal and sublingual) or transdermal administration of the compounds of the present technology include powders, sprays, ointments, pastes, creams, lotions, gels, solutions and patches. The active component can be mixed with a pharmaceutically acceptable carrier or excipient and any preservative or buffer that may be needed under aseptic conditions. Powders and sprays can be prepared, for example, with excipients such as lactose, talcum, silicic acid, aluminum hydroxide, calcium silicate and polyamide powders or mixtures of these substances. Ointments, pastes, creams and gels can also contain excipients such as animal and vegetable fats, oils, waxes, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicones, bentonite, silicic acid, talcum and zinc oxide or mixtures thereof. Absorption enhancers can also be used to increase the flux of the compounds of the present technology through the skin. The rate of such flux can be controlled by providing a rate-controlling membrane (for example, as part of a transdermal patch) or by dispersing the compound in a polymer matrix or gel.

[0099] In addition to those representative dosage forms described above, suitable pharmaceutically acceptable excipients and carriers are generally known to those skilled in the art and are therefore included in the prior art. Such excipients and carriers are described in, for example, "Remingtons Pharmaceutical Sciences" Mack Pub. Co., New Jersey (1991), which is incorporated herein by reference.

[0100] The formulations of the present technology can be designed to be short-acting, fast-releasing, long-acting, and sustained-releasing, as described below. Therefore, the pharmaceutical formulations can also be formulated for controlled or sustained release.

[0101] The compositions of the present invention may also comprise, for example, micelles or liposomes, or some other encapsulation forms, or may be administered in a sustained release form to provide extended storage and / or delivery effects. Thus, pharmaceutical preparations and drugs may be compressed into pills or cylinders and implanted intramuscularly or subcutaneously as reservoir injections or as implants (such as stents). Such implants may be made of known inert materials, such as silicone and biodegradable polymers.

[0102] Specific dosage can be adjusted according to the disease state, age, weight, general health, sex and diet of the subject, dosage interval, administration route, excretion rate and drug combination. Any of the above dosage forms containing an effective amount is completely within the scope of routine experiments, and therefore is completely within the scope of the existing present technology.

[0103] Those skilled in the art can easily determine the effective amount by simply administering the compound of the present technology to the patient in an increasing amount until (for example) the tumor mass of the subject is reduced. The compound of the present technology can be administered to the patient at a dosage level ranging from about 0.1 mg to about 1,000 mg per day. For a normal adult weighing about 70 kg, a dosage ranging from about 0.01 mg to about 100 mg per kg of body weight per day is sufficient. However, the specific dosage used may vary or may be adjusted as appropriate by a person of ordinary skill in the art. For example, the dosage may depend on many factors, including the patient's needs, the severity of tumor-related B-cell malignancies (e.g., non-Hodgkin lymphoma or chronic lymphocytic leukemia), and the pharmacological activity of the compound used. The determination of the optimal dosage for a particular patient is well known to those skilled in the art.

[0104] Various assays and model systems can be readily employed to determine the therapeutic efficacy of treatments according to the present technology. The effectiveness of the compositions (and determination of effective amounts) and methods of the present technology can also be demonstrated by the following: reduction in tumor mass and / or slowing tumor growth and / or affecting the increase in the therapeutic responsiveness of cancer to a second cancer therapy (such as radiotherapy, monoclonal antibodies and / or chemotherapeutic agents).

[0105] For each specified condition described herein, the test subject will exhibit a 10%, 20%, 30%, 50% or greater reduction, up to a 75% to 90% or 95% or greater reduction, in one or more symptoms caused by or associated with the subject's condition, compared to a placebo-treated control subject or other appropriate control subject.

[0106] The compounds of the present technology can also be applied to patients together with other conventional therapeutic agents that can be used to treat tumors or vaccinations. Administration can include oral administration, parenteral administration or nasal administration. In any of these embodiments, administration can include intratumoral injection, subcutaneous injection, intravenous injection, intraperitoneal injection or intramuscular injection. In any of these embodiments, administration can include oral administration. The method of the present technology can also include conventional therapeutic agents and one or more compounds of the present technology in an amount that can potentially or synergistically effectively treat cancer (e.g., breast cancer, ovarian cancer, pancreatic cancer, biliary tract cancer, lung cancer, prostate cancer and / or CNS cancer) sequentially or in combination.

[0107] In one aspect, the compounds of the present technology are administered to a patient in an amount or dosage suitable for therapeutic use. In general, the unit dose comprising the compounds of the present technology will vary according to the patient's considerations. These considerations include, for example, age, regimen, condition, gender, extent of disease, contraindications, concomitant therapy, and the like. Exemplary unit doses based on these considerations can also be adjusted or modified by those skilled in the art. For example, a unit dose for a patient comprising a compound of the present technology can be in the range of 1×10 - 4 g / kg to 1 g / kg, preferably 1×10 -3 The dosage of the compounds of the present technology may also vary between 0.01 mg / kg and 100 mg / kg, or preferably between 0.1 mg / kg and 10 mg / kg.

[0108] The compounds of the present technology can also be modified, for example, by covalently linking an organic moiety or conjugate to improve pharmacokinetic properties, toxicity or bioavailability (e.g., increase in vivo half-life). The conjugate can be a linear or branched hydrophilic polymer group, a fatty acid group or a fatty acid ester group. The polymer group can include a molecular weight that can be adjusted by a person of ordinary skill in the art to improve, for example, pharmacokinetic properties, toxicity or bioavailability. Exemplary conjugates can include polyalkylene glycols (e.g., polyethylene glycol (PEG), polypropylene glycol (PPG)), carbohydrate polymers, amino acid polymers or polyvinyl pyrrolidone and fatty acid or fatty acid ester groups, each of which can independently contain about eight to about seventy carbon atoms. The conjugates used with the compounds of the present technology can also be used as connectors to, for example, any suitable substituent or group, radiolabel (marker or label), halogen, protein, enzyme, polypeptide, other therapeutic agent (e.g., medicine or drug), nucleoside, dye, oligonucleotide, lipid, phospholipid and / or liposome. In one aspect, the conjugate can include polyethyleneamine (PEI), polyglycine, a hybrid of PEI and polyglycine, polyethylene glycol (PEG) or methoxypolyethylene glycol (mPEG). The conjugate can also connect the compound of the present technology to (for example) a label (fluorescent label or luminescent label) or a marker (radionuclide, radioisotope and / or isotope) to form a probe of the present technology. The conjugate used with the compound of the present technology can improve the half-life in vivo in one aspect. Other exemplary conjugates used with the compound of the present technology and their applications and related technologies include those summarized in U.S. Patent No. 5,672,662, which is hereby incorporated by reference herein.

[0109] In another aspect, the present technology provides a method for identifying a target of interest, comprising contacting the target of interest with a detectable or imaging effective amount of a labeled compound of the present technology. A detectable or imaging effective amount is the amount of the labeled compound of the present technology necessary to be detected by the selected detection method. For example, a detectable amount can be an amount of administration sufficient to detect the binding of the labeled compound to the target of interest. Suitable labels are known to those skilled in the art and may include, for example, radioisotopes, radionuclides, isotopes, fluorescent groups, biotin (bound to streptavidin complexes), and chemiluminescent groups. After the labeled compound is bound to the target of interest, the target can be separated, purified, and further characterized (such as by determining the amino acid sequence).

[0110] The terms "association" and / or "binding" may refer to, for example, a chemical or physical interaction between a compound of the present technology and a target of interest. Examples of associations or interactions include covalent bonds, ionic bonds, hydrophilic-hydrophilic interactions, hydrophobic-hydrophobic interactions, and complexes. Association may also generally refer to "binding" or "affinity," as each can be used to describe a variety of chemical or physical interactions. Measuring binding or affinity is also routine for those skilled in the art. For example, a compound of the present technology may bind to or interact with a target of interest or a precursor, portion, fragment, and peptide thereof, and / or their deposits.

[0111] As previously noted in the present disclosure, in one aspect, a method of treating a subject suffering from cancer is provided, wherein the method comprises administering to the subject an effective amount of a compound of any embodiment disclosed herein or administering an effective amount of a composition of any embodiment disclosed herein, and optionally an effective amount of a second cancer therapy. In any embodiment herein, the cancer can be breast cancer, ovarian cancer, pancreatic cancer, biliary tract cancer, lung cancer, prostate cancer, and / or CNS cancer (such as brain cancer).

[0112] In any embodiment herein, administration may further include administration of radiation therapy, monoclonal antibodies and / or chemotherapeutic agents (such as alkylating agents; nitrosoureas; antimetabolites; anthracyclines; topoisomerase II inhibitors; mitotic inhibitors; or antiestrogens; progestins; aromatase inhibitors; antiandrogens; LHRH agonists; corticosteroids; DNA alkylating agents; taxanes; vinca alkaloids; microtubule toxins, or any two or more thereof. In any embodiment herein, administration may also include administration of chemotherapeutic agents such as busulfan, cisplatin, carboplatin, oxaliplatin, octahedral platinum (IV) compounds, chlorambucil, cyclophosphamide, ifosfamide, dacarbazine (DTIC), dichloromethyl diethylamide (nitrogen mustard), melphalan, temozolomide, carmustine (BCNU), lomustine (CCNU), 5-fluorouracil, capecitabine, 6 -Mercaptopurine, methotrexate, gemcitabine, cytarabine (ara-C), fludarabine, pemetrexed, daunorubicin, doxorubicin (Adriamycin), epirubicin, idarubicin, mitoxantrone, topotecan, irinotecan, etoposide (VP-16), teniposide, paclitaxel, docetaxel, vinblastine, vincristine, vinorelbine, prednisone, dexamethasone, L-asparaginase, actinomycin, thalidomide , tretinoin, imatinib (Gleevec), gefitinib (Iressa), erlotinib (Tarceva), rituximab (MabThera), bevacizumab (Avastin), ipilimumab, nivolumab (Opdivo), pembrolizumab (Keytruda), tamoxifen, fulvestrant, anastrozole, exemestane, letrozole, megestrol acetate, bicalutamide, flutamide, leuprolide, goserelin, or a combination of any two or more thereof.

[0113] In any embodiment herein, administration may include oral, rectal, nasal, vaginal, parenteral, transdermal, intravenous, intramuscular or inhalation administration. In any embodiment herein, administration may include topical administration of a compound to a site where the subject includes cancer or topical administration of a composition to a site where the subject includes cancer.

[0114] The examples herein are provided to demonstrate the advantages of the present technology and to further assist those of ordinary skill in the art in preparing or using the compounds and compositions of the present technology. Examples herein are also presented to more fully illustrate the preferred aspects of the present technology. Examples should never be interpreted as limiting the scope of the present technology as defined by the appended claims. Examples may include or incorporate any variant, aspect or embodiment of the present technology described above. The variations, aspects or embodiments described above may also further include or incorporate any or all other variations, aspects or embodiments of the present technology.

[0115] Examples

[0116] Unless otherwise stated, all solvents and reagents were used as received from commercial suppliers. 1 H spectra were obtained on a Bruker AM or Varian 400 spectrometer (operating at 400 MHz, respectively) in CDCl 3 The chemical shifts (δ) are reported in parts per million (ppm) and the coupling constants (J) are reported in Hertz (Hz). Spin multiplicities are reported as s = singlet, d = doublet, t = triplet, q = quartet, dd = doublet of doublet, ddd = doublet of doublet of doublet, dt = doublet of triplet, td = triplet of doublet, and m = multiplet. LCMS analysis was performed on a chromatograph with photodiode array UV detection and a TOF mass spectrometer. The mass spectrometer utilizes a multimode source with simultaneous ESI+ / APCI+ acquisition; a reference mass solution; and a supplemental solvent introduced into the LC flow prior to the source to assist ionization. Although single stereoisomers were isolated and obtained in high purity in the following examples, the final unambiguous attribution of absolute stereochemistry may not have been determined for some groups of stereoisomers.

[0117] Comparative Synthesis of Compounds 0092A and 0092B:

[0118]

[0119] Step 1: To a solution of 6-fluoro-4-nitro-3H-2-benzofuran-1-one (400 mg, 2.03 mmol) in THF (20 mL) was added TEA (575 mg, 5.68 mmol) and Ac 2 O (1284 mg, 12.58 mmol) and 2-fluoro-4-formylbenzonitrile (605 mg, 4.06 mmol). The solution was stirred at 80 ° C for 3 h. The reaction solution was cooled to room temperature. The mixture was filtered. The filter cake was concentrated in vacuo to obtain a yellow solid crude product (290 mg, 41%), which was used directly without further purification. MS (ESI): C 16 H 6 F 2 N 2 O 4 Calculated mass, 328.0, observed m / z, 351.0 [M+Na]+

[0120] Step 2: To a solution of 2-fluoro-4-{[(1Z)-5-fluoro-7-nitro-3-oxo-2-benzofuran-1-ylidene]methyl}benzonitrile (300 mg, 0.91 mmol) in MeOH (10 mL) was added HCl (4N in MeOH, 10 mL). The solution was stirred at 25 °C for 12 h. The solvent was removed in vacuo. The crude product (300 mg, 87%) was used directly in the next step without further purification. MS (ESI): C 17 H 10 F 2 N 2 O 5 Calculated mass, 360.0, observed m / z 361.0 [M+H]+

[0121] Step 3: To a solution of methyl 2-[2-(4-cyano-3-fluorophenyl)acetyl]-5-fluoro-3-nitrobenzoate (200 mg, 0.56 mmol) and 2,4,6-trifluorobenzaldehyde (166 mg, 1.04 mmol) in THF (30 mL) and MeOH (5 mL) was added TiCl 3 (20% HCl solution, 2548 mg, 3.33 mmol). The solution was stirred at 40 °C for 12 h. The mixture was washed with H 2 O (50 mL), extracted with EA (50 mL * 3), the combined organic layer was washed with brine (50 mL), and then purified by Na 2 SO 4 Dry and concentrate in vacuo. The crude product was purified by preparative TLC (PE:EA=3:1) to give the desired product (230 mg, 84%) as a white solid. MS (ESI): C 24 H 13 F 5 N 2 O 3 Calculated mass, 472.0, observed m / z 473.0 [M+H]+

[0122] Step 4: To a solution of methyl 3-(4-cyano-3-fluorophenyl)-7-fluoro-4-oxo-2-(2,4,6-trifluorophenyl)-2,3-dihydro-1H-quinoline-5-carboxylate (100 mg, 0.21 mmol) in MeOH (10 mL) was added N 2 H 4 .H 2 O (37 mg, 0.74 mmol). The solution was stirred at 25 °C for 2 h. 2 O (50 mL), extracted with EA (50 mL * 3), and the combined organic layer was washed with brine (50 mL). 2 SO4 Dry and concentrate in vacuo. The crude product is purified by flash (CH 3 CN:H 2 O=1:3) to obtain the desired product (50 mg, 51%) as a white solid. MS (ESI): C 23 H 11 F 5 N 4 Calculated mass of O, 454.0, observed m / z 455.0 [M+H]+

[0123] Step 5: 2-Fluoro-4-(5-fluoro-3-oxo-8-(2,4,6-trifluorophenyl)-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]naphthyridin-9-yl)benzonitrile was purified by preparative SFC (apparatus: SFC 80; column: Daicel CHIRALCELOD, 250 mm×30 mm ID, 10 μm; mobile phase: CO 2 / MeOH[0.2% NH 3 (7M MeOH solution)] = 50 / 50; flow rate: 70 g / min; wavelength: UV 214 nm; temperature: 35°C) to give white solid 2-fluoro-4-((8S,9R)-5-fluoro-3-oxo-8-(2,4,6-trifluorophenyl)-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-9-yl)benzonitrile (13.1 mg, 51%) and white solid 2-fluoro-4-((8R,9S)-5-fluoro-3-oxo-8-(2,4,6-trifluorophenyl)-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-9-yl)benzonitrile (12.6 mg, 51%). MS (ESI): C 23 H 11 F 5 N 4 The mass calculated value of O is 454.0, and the experimental value of m / z is 455.0 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 12.33 (s, 1H), 7.89 (s, 1H), 7.81 (t, J = 8.0 Hz, 1H), 7.60 (d, J = 12.0 Hz, 1H), 7.31 (dd, J = 8.0 Hz, 1.2 Hz, 1H), 7.18 (t, J = 8.0 Hz, 2H), 7.09 (dd, J = 9.2 Hz, 1.6 Hz, 1H), 6.85 (dd, J = 10.8 Hz, 2.4 Hz, 1H), 5.28 (d, J = 12.0 Hz, 1H), 4.71 (d, J = 12.0 Hz, 1H).

[0124] Synthesis of 0110A and 0110B

[0125]

[0126] Step 1: In N 2 To a solution of 6-fluoro-4-nitro-3H-2-benzofuran-1-one (5 g, 25.4 mmol) in THF (80 mL) were added trimethylamine (5.14 g, 50.8 mmol), Ac 2 O (15 mL) and 3,4-difluorobenzaldehyde (7.22 g, 50.80 mmol). The reaction mixture was stirred at 25 ° C for 5 min, and then warmed to 80 ° C and stirred for 3 h. The reaction mixture was quenched with water (100 mL) and extracted with EtOAc (100 mL×3). The combined organic layer was washed with brine (100 mL) and purified by Na 2 SO 4 Drying and concentration gave a residue. The residue was stirred with EtOAc (30 mL), filtered and the filter cake was collected and concentrated to give (Z)-3-(2,4-difluorobenzylidene)-6-fluoro-4-nitroisobenzofuran-1(3H)-one (2.2 g, 24%) as a yellow solid.

[0127] Step 2: (Z)-3-(2,4-difluorobenzylidene)-6-fluoro-4-nitroisobenzofuran-1(3H)-one (1.0 g, 3.00 mmol) was added to a 4M HCl solution in MeOH (30 mL). The reaction mixture was stirred at 70 °C for 48 h. The reaction mixture was concentrated to give methyl 2-(2-(2,4-difluorophenyl)acetyl)-5-fluoro-3-nitrobenzoate (1.1 g, 90%) as a yellow solid. MS (ESI): C 16 H 10 F 3 NO 5 Calculated mass, 353.05, m / z observed, 354.1 [M+H] + .

[0128] Step 3: In N 2 To a solution of methyl 2-(2-(2,4-difluorophenyl)acetyl)-5-fluoro-3-nitrobenzoate (270 mg, 0.76 mmol) and 1-methylpiperidine-4-carbaldehyde (194 mg, 1.50 mmol) in THF (18 mL) and MeOH (3 mL) was added TiCl at 0 °C. 3 HCl solution (5.25 g, 6.82 mmol). The reaction mixture was stirred at 40 °C for 16 h. The reaction mixture was quenched with water (40 mL) and extracted with EtOAc (60 mL×3). The combined organic layers were washed with brine (60 mL) and purified by Na 2 SO4 Drying and concentration gave a residue. The residue was purified by preparative TLC (MeOH / DCM=1 / 10, rf=0.4), and the fractions with the MS signal of the desired product were collected and concentrated to give 3-(2,4-difluorophenyl)-7-fluoro-2-(1-methylpiperidin-4-yl)-4-oxo-1,2,3,4-tetrahydroquinoline-5-carboxylic acid methyl ester (60 mg, 18%) as a yellow solid. MS (ESI): C 23 H 23 F 3 N 2 O 3 Calculated mass, 432.17, observed m / z 433.1 [M+H] + .

[0129] Step 4: A solution of methyl 3-(2,4-difluorophenyl)-7-fluoro-2-(1-methylpiperidin-4-yl)-4-oxo-1,2,3,4-tetrahydroquinoline-5-carboxylate (60 mg, 0.18 mmol) in MeOH (8 mL) was stirred at 25 °C for 20 min, and N 2 H 4 .H 2 O (436 mg, 6.96 mmol). The mixture was stirred at 25 ° C for 16 h. The reaction mixture was concentrated to obtain a residue. The residue was purified by preparative HPLC (chromatographic column: Xbridge 5u-C18 150×19 mm, 5 μm mobile phase: ACN--H 2 O (0.1% formic acid), B (acetonitrile); flow rate: 20 mL / min, wavelength: 214 / 254 nm) to obtain 9-(2,4-difluorophenyl)-5-fluoro-8-(1-methylpiperidin-4-yl)-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one (40 mg, 50%) as a yellow solid. MS (ESI): C 22 H 21 N 3 O 4 Calculated mass, 414.17, observed m / z 415.1 [M+H] + .

[0130] Step 5: 9-(2,4-difluorophenyl)-5-fluoro-8-(1-methylpiperidin-4-yl)-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one (40 mg, 0.09 mmol) was purified by preparative SFC (Daicel CHIRALPAK IB 30×250 mm, 10 μm mobile phase A / B: CO 2 / MeOH (0.1% EDA)) = 60 / 40; flow rate: 1.5 mL / min, column temperature: 34 degrees) to obtain (8R, 9R)-9-(2,4-difluorophenyl)-5-fluoro-8-(1-methylpiperidin-4-yl)-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one (6.7 mg, 17%) as a white solid and (8R, 9S)-9-(2,4-difluorophenyl)-5-fluoro-8-(1-methylpiperidin-4-yl)-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one (8.2 mg, 20%) as a white solid.

[0131] 0110A: MS (ESI): mass calculated for C22H21F3N4O, 414.17, m / z observed 415.1 [M+H] + . 1 HNMR (400MHz, MeOD) δ = 7.15 (dd, J = 10.0Hz, 4.0Hz, 1H), 7.03-6.98 (m, 1H), 6.85-6.78 (m, 2H), 6.75-6.69 (m, 1H), 4.55 (d, J = 3 .6Hz,1H),3.37-3.31(m,1H),2.99-2.90(m,2H),2.33(s,3H),2.14-2.03(m,2H),1.81(d,J=12.0Hz,1H),1.63-1.39(m,4H).

[0132] 0110B: MS (ESI): mass calculated for C22H21F3N4O, 414.17, m / z observed 415.1 [M+H] + . 1 HNMR (400MHz, MeOD) δ = 7.14 (dd, J = 10.0Hz, 4.0, 1H), 7.02-6.97 (m, 1H), 6.85- 6.78(m,2H),6.75-6.69(m,1H),4.55(d,J=3.6Hz,1H),3.36-3.35(m,1H),2.99 -2.90(m,2H),2.27(s,3H),2.06-1.95(m,2H),1.78(d,J=12.0Hz,1H),1.53-1.41(m,4H).

[0133] Synthesis of 0111-6, 0111, 0111B, 0111C and 0111D:

[0134]

[0135] Step 1: To a mixture of methyl 2-[2-(2,4-difluorophenyl)acetyl]-5-fluoro-3-nitrobenzoate (1.5 g, 4.20 mmol) and tert-butyl (3-formylpyrrolidin-1-yl)carboxylate (1.7 g, 8.40 mmol) in MeOH (4 mL) and THF (24 mL) was added TiCl 3 (15.0 g, 25.20 mmol). The reaction mixture was stirred at 40 °C for 16 h. 2 O (50 mL) and extracted with EA (2×50 ml). The organic layer was washed with brine (50 mL) and washed with Na 2 SO 4 Dry, filter and concentrate the filtrate to give a residue which is purified by reverse phase column (30% A in B; A: CH 3 CN, B: 0.1% TFA aqueous solution) to give 3-(2,4-difluorophenyl)-7-fluoro-4-oxo-2-(pyrrolidin-3-yl)-2,3-dihydro-1H-quinoline-5-carboxylic acid methyl ester (0.7 g, 38%) as a yellow oil. MS (ESI): C 21 H 19 F 3 N 2 O 3 Calculated mass, 404.13, observed m / z 405.0 [M+H] + .

[0136] Step 2: To a solution of methyl 3-(2,4-difluorophenyl)-7-fluoro-4-oxo-2-(pyrrolidin-3-yl)-2,3-dihydro-1H-quinoline-5-carboxylate (950 mg, 2.35 mmol) in ACN (10 mL) were added 1-bromo-2-methoxyethane (327 mg, 2.35 mmol) and K 2 CO 3 (974 mg, 7.05 mmol). The reaction mixture was stirred at 80 °C for 16 h. After cooling to room temperature, the reaction mixture was washed with H 2 O (15 mL) and extracted with EA (2×20 mL). The organic layer was washed with brine (20 mL) and washed with anhydrous Na 2 SO 4 Dry and concentrate in vacuo. The residue was purified by flash chromatography on a silica gel column (PE:EA=3:1) to give methyl 3-(2,4-difluorophenyl)-7-fluoro-2-[1-(2-methoxyethyl)pyrrolidin-3-yl]-4-oxo-2,3-dihydro-1H-quinoline-5-carboxylate (600 mg, 50%) as a yellow oil. MS (ESI): C 24 H 25 F 3N 2 O 4 Calculated mass, 462.18, observed m / z 463.1 [M+H] + .

[0137] Step 3: To a solution of methyl 3-(2,4-difluorophenyl)-7-fluoro-2-[1-(2-methoxyethyl)pyrrolidin-3-yl]-4-oxo-2,3-dihydro-1H-quinoline-5-carboxylate (600 mg, 1.30 mmol) in MeOH (10 mL) was added N 2 H 4 H 2 O (650 mg, 12.97 mmol). The reaction mixture was stirred at 25 °C for 16 h. 2 O (15 mL) and extracted with EA (2×20 mL). The organic layer was washed with brine (20 mL) and washed with anhydrous Na 2 SO 4 The residue was purified by preparative HPLC (instrument: Waters Prep-HPLC, column: Xbridge preparative c18 5 μm OBD 19×150 mm / WELCH Xtimate C18 21.2×250 mm 10 μm, A water (0.1% formic acid), B acetonitrile, 10-20% B in 8 min, at 100% B for 2 min, return to 5% B in 0.5 min, stop at 13 min; flow rate: 20 ml / min, wavelength: 214 / 254 nm) to obtain 12-(2,4-difluorophenyl)-7-fluoro-11-[1-(2-methoxyethyl)pyrrolidin-3-yl]-2,3,10-triazatricyclo[7.3.1.0^{5,13}]trideca-1,5(13),6,8-tetraen-4-one ("0111-6"; 300 mg, 50%) as a yellow solid. MS (ESI): C 23 H 23 F 3 N 4 O 2 Calculated mass, 444.18, observed m / z 445.1 [M+H] + . 1 H NMR (400MHz, CD 3OD)δ8.36(s,1H),7.24-7.15(m,1H),7.08-6.96(m,1H),6.91-6.62(m,3H),4.47-4.27(m,1H),3.71-3.5 7(m,4H),3.45-3.39(m,3H),3.39-3.33(m,3H),3.29-3.23(m,1H),2.75-2.59(m,1H),2.39-1.96(m,3H).

[0138] Step 4: 0111-6 (300 mg, 0.68 mmol) was separated by preparative SFC (device: SFC 80 chromatographic column: Daicel CHIRALPAK OX_3, 3×150 mm, 3 μm, mobile phase: A / B: CO 2 / MeOH (0.1% DEA) = 65 / 35; flow rate: 70 g / min, wavelength: UV 214 nm, temperature: 35°C) and (device: SFC 80 chromatographic column: Daicel CHIRALPAK IE_3, 3.0×150 mm, 3 μm, mobile phase: A / B: CO 2 / MeOH (0.1% DEA) = 65 / 35; flow rate: 70 g / min, wavelength: UV214 nm, temperature: 35°C) to obtain a yellow solid 0111 (27.2 mg, 9%), a white solid 0111B (20.7 mg, 7%), a white solid 0111C (48 mg, 16%) and a yellow solid 0111D (54.9 mg, 18%).

[0139] 0111A: 1 H NMR (400 MHz, CDCl 3 )δ9.86(s,1H),7.32-7.27(m,1H),7.00-6.82(m,3H),6.75-6.69(m,1H),6.34(s,1H),4.22(d,J=8.0Hz,1H),3.60-3.48(m,3H),3.37(s,3H ), 3.04 (d, J = 8.0Hz, 2H), 2.82-2.72 (m, 1H), 2.70-2.60 (m, 1H), 2.43-2.30 (m, 2H), 2.29-2.20 (m, 1H), 2.11-2.01 (m, 1H), 1.82-1.70 (m, 1H).

[0140] 0111B: 1 H NMR (400 MHz, CDCl 3)δ 9.74 (s, 1H), 7.31 - 7.27 (m, 1H), 7.01 - 6.92 (m, 1H), 6.91 - 6.83 (m, 2H), 6.77 - 6.69 (m, 1H), 6.37 (s, 1H), 4.21 (d, J = 9.1 Hz, 1H), 3.61 - 3.49 (m, 3H), 3.37 (s, 3H), 3.14 - 3.00 (m, 2H), 2.84 - 2.73 (m, 1H), 2.73 - 2.63 (m, 1H), 2.42 - 2.31 (m, 2H), 2.30 - 2.20 (m, 1H), 2.12 - 2.02 (m, 1H), 1.83 - 1.74 (m, 1H).

[0141] 0111C: 1 H NMR (400 MHz, CDCl 3 )δ 9.65 (s, 1H), 7.26 - 7.22 (m, 2H), 7.16 - 7.08 (m, 1H), 6.97 - 6.84 (m, 2H), 6.73 - 6.63 (m, 1H), 4.13 (d, J = 12.0 Hz, 1H), 3.77 (d, J = 12.0 Hz, 1H), 3.60 - 3.52 (m, 2H), 3.43 (s, 3H), 3.27 - 3.14 (m, 1H), 3.04 - 2.90 (m, 1H), 2.84 - 2.74 (m, 1H), 2.69 - 2.56 (m, 1H), 2.44 - 2.30 (m, 1H), 2.28 - 2.17 (m, 1H), 2.10 - 2.02 (m, 1H), 1.92 - 1.75 (m, 2H).

[0142] 0111D: 1 H NMR (400 MHz, CDCl 3 )δ 9.70 (s, 1H), 7.26 - 7.21 (m, 2H), 7.17 - 7.07 (m, 1H), 7.00 - 6.77 (m, 2H), 6.65 (d, J = 12.0 Hz, 1H), 4.13 (d, J = 12.0 Hz, 1H), 3.77 (d, J = 12.0 Hz, 1H), 3.60 - 3.49 (m, 2H), 3.44 (s, 3H), 3.26 - 3.11 (m, 1H), 3.02 - 2.88 (m, 1H), 2.85 - 2.69 (m, 1H), 2.69 - 2.53 (m, 1H), 2.45 - 2.27 (m, 1H), 2.25 - 1.98 (m, 3H), 1.90 - 1.77 (m, 1H).

[0143] Synthesis of 0114A and 0114B

[0144]

[0145] Step 1: In N 2 To a solution of methyl 2-(2-(2,4-difluorophenyl)acetyl)-5-fluoro-3-nitrobenzoate (2 g, 5.67 mmol) in THF / MeOH=6 / 1 (14 mL) was added paraformaldehyde (1.5 g, 17.01 mmol) and TiCl 3 HCl solution (14 mL) was added, and the reaction mixture was stirred at 40°C for 6 h. The reaction mixture was quenched with water (50 mL) and extracted with EtOAc (50 mL×3), and the combined organic layers were washed with brine (80 mL) and purified by Na 2 SO 4 The residue was dried and concentrated to give a residue. The residue was purified by column chromatography on silica gel (25 g) eluting with petroleum ether (45%) containing EtOAc, and the fractions with the MS signal of the desired product were collected and concentrated to give methyl 3-(2,4-difluorophenyl)-7-fluoro-4-oxo-1,2,3,4-tetrahydroquinoline-5-carboxylate (150 mg, 7.9%) as a yellow oil. MS (ESI): C 17 H 12 F 3 NO 3 Calculated mass, 335.08, observed m / z 336.1 [M+H] + .

[0146] Step 2: In N 2 To a solution of methyl 3-(2,4-difluorophenyl)-7-fluoro-4-oxo-2,3-dihydro-1H-quinoline-5-carboxylate (150 mg, 0.45 mmol) in MeOH (8 mL) was added N 2 H 4 .H 2 O (8 mL), and the reaction mixture was stirred at 25 ° C for 16 h. The reaction mixture was directly concentrated to obtain a residue. The residue was purified by preparative HPLC (chromatographic column: Xbridge 5u-C18 150×19 mm, 5 μm mobile phase A: ACN--H 2 O (0.1% formic acid), B (acetonitrile); flow rate: 20 mL / min, wavelength: 214 / 254 nm) to obtain 9-(2,4-difluorophenyl)-5-fluoro-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one (60 mg, 42%) as a white solid. MS (ESI): C 16 H 10 F 3 N 3 Calculated mass of O, 317.08, observed m / z 318.1 [M+H]+ .

[0147] Step 3: 9-(2,4-difluorophenyl)-5-fluoro-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one (60 mg, 0.19 mmol) was separated by preparative SFC (Daicel CHIRALPAK AD_3 3×150 mm, 3 μm mobile phase A / B: CO 2 / MeOH (0.1% EDA)) = 65 / 35; flow rate: 2.0 mL / min, column temperature: 37 degrees) to obtain white solid (R)-9-(2,4-difluorophenyl)-5-fluoro-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one (24.0 mg, 40%) and white solid (S)-9-(2,4-difluorophenyl)-5-fluoro-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one (25.6 mg, 43%). MS (ESI): C 16 H 10 F 3 N 3 Calculated mass of O, 317.08, observed m / z 318.1 [M+H] + .

[0148] 0114A: 1 H NMR (400MHz, CD 3 OD)δ7.18(dd,J=9.2,2.4Hz,1H),7.10-6.94(m,2H),6.94-6.75(m,2H),4.52-4.47(m,1H),3.71-3.57(m,2H).

[0149] 0114B: 1 H NMR (400MHz, CD 3 OD)δ7.18(dd,J=9.2,2.4Hz,1H),7.10-6.95(m,2H),6.93-6.78(m,2H),4.53-4.47(m,1H),3.71-3.56(m,2H).

[0150] Synthesis of 0115A, 0115B, 0115C and 0115D:

[0151]

[0152] Step 1: To a solution of methyl 2-[2-(2,4-difluorophenyl)acetyl]-5-fluoro-3-nitrobenzoate (950 mg, 2.68 mmol) in THF (15 mL) and MeOH (3 mL) was added TiCl under ice bath. 3 (12.7 g, 16.13 mmol). The reaction mixture was stirred at 40 °C for 2 h. Then oxacyclopentane-3-carboxaldehyde (808 mg, 8.06 mmol) was added under ice bath. The mixture was stirred at 25 °C for 14 h. The reaction mixture was washed with H 2 O (50 mL) and extracted with EA (2×50 ml). The organic layer was washed with brine (50 mL) and purified by Na 2 SO 4 The residue was dried, filtered, and concentrated to give a residue, which was purified by silica gel column chromatography using PE:EA (3:1) as eluent to give yellow solid product compound 3 (200 mg) and yellow solid compound 3a (200 mg). MS (ESI): C 21 H 18 F 3 NO 4 Calculated mass, 405.12, observed m / z 406.1 [M+H] + .

[0153] Step 2: To a solution of methyl (2R,3R)-3-(2,4-difluorophenyl)-7-fluoro-4-oxo-2-(oxolan-3-yl)-2,3-dihydro-1H-quinoline-5-carboxylate (200 mg, 0.49 mmol) in MeOH (5 mL) was added NH 2 NH 2 .H 2 O (1.56 g, 24.67 mmol). The reaction mixture was stirred at 25 °C for 3 h. 2 O (15 mL) and extracted with EA (2×20 mL). The organic layer was washed with brine (20 mL) and washed with anhydrous Na 2 SO 4 The residue was purified by silica gel column chromatography using PE / EA (2:1) as eluent to give the product (110 mg). The product was then separated by preparative SFC (device: SFC80, column: Daicel CHIRALCEL OJ, 250 mm × 30 mm ID, 10 μm, mobile phase: CO 2 / MeOH[0.2% NH 3(7 M MeOH solution)] = 80 / 20; flow rate: 70 g / min, wavelength: UV 214 nm, temperature: 35°C), thereby obtaining 25.3 mg of a white solid product (0115) and 24.0 mg of a white solid product (0115B).

[0154] 0115A:MS(ESI):C 20 H 16 F 3 N 3 O 2 Calculated mass, 387.12, observed m / z 388.1 [M+H] + . 1 HNMR(400MHz,MeOD)δ7.16(dd,J=8.8,2.0Hz,1H),7.02-6.97(m,1H),6.90–6.72(m,3H),4.27(d,J=4.0 Hz,1H),3.98–3.83(m,2H),3.75-3.62(m,2H),3.56-3.50(m,1H),2.50-2.39(m,1H),2.05-1.82(m,2H).

[0155] 0115B:MS(ESI):C 20 H 16 F 3 N 3 O 2 Calculated mass, 387.12, observed m / z 388.1 [M+H] + . 1 HNMR(400MHz,MeOD)δ7.16(dd,J=8.8,2.0Hz,1H),7.02-6.96(m,1H),6.90–6.72(m,3H),4.27(d,J=4.0 Hz,1H),3.96–3.84(m,2H),3.75-3.62(m,2H),3.56-3.50(m,1H),2.50-2.39(m,1H),2.05-1.82(m,2H).

[0156] Step 3: To a solution of methyl (2S,3S)-3-(2,4-difluorophenyl)-7-fluoro-4-oxo-2-(oxolan-3-yl)-2,3-dihydro-1H-quinoline-5-carboxylate (200 mg, 0.49 mmol) in MeOH (5 mL) was added NH 2 NH 2 .H 2 O (1.56 g, 24.67 mmol). The reaction mixture was stirred at 25 °C for 3 h. 2O (15 mL) and extracted with EA (2×20 mL). The organic layer was washed with brine (20 mL) and washed with anhydrous Na 2 SO 4 The residue was purified by silica gel column chromatography using PE / EA (2:1) as eluent to give the product (120 mg). The product was then separated by preparative SFC (device: SFC80, column: Daicel CHIRALCEL IC, 250 mm × 30 mm ID, 10 μm, mobile phase: CO 2 / MeOH [0.2% NH 3 (7 M MeOH solution)] = 65 / 35; flow rate: 70 g / min, wavelength: UV 214 nm, temperature: 35°C), thereby producing 35.4 mg (0115C) of a white solid product and 30.6 mg (0115D) of a white solid product.

[0157] 0115C:MS(ESI):C 20 H 16 F 3 N 3 O 2 Calculated mass, 387.12, observed m / z 388.1 [M+H] + . 1 HNMR(400MHz,MeOD)δ7.16(dd,J=9.2,2.4Hz,1H),7.05–6.95(m,1H),6.85-6.75(m,2H),6.70-6.62(m,1H),4.45(d,J=3.2 Hz,1H),3.99-3.91(m,1H),3.80-3.66(m,3H),3.50-3.44(m,1H),2.54-2.40(m,1H),2.20-2.09(m,1H),1.90-1.78(m,1H).

[0158] 0115D:MS(ESI):C 20 H 16 F 3 N 3 O 2 Calculated mass, 387.12, observed m / z 388.1 [M+H] + . 1HNMR(400MHz,MeOD)δ7.16(dd,J=9.2,2.4Hz,1H),7.05–6.96(m,1H),6.85-6.75(m,2H),6.70-6.62(m,1H),4.45(d,J=3.2 Hz,1H),3.99-3.92(m,1H),3.80-3.66(m,3H),3.50-3.44(m,1H),2.54-2.40(m,1H),2.20-2.09(m,1H),1.90-1.78(m,1H).

[0159] Synthesis of 0116A and 0116B

[0160]

[0161] Step 1: In N 2 To a solution of 6-fluoro-4-nitro-3H-2-benzofuran-1-one (1 g, 5.10 mmol) in THF (20 mL) were added trimethylamine (1.44 g, 14.20 mmol), Ac 2 O (3.23 g, 31.60 mmol) and 2-chlorobenzaldehyde (1.43 g, 10.20 mmol). The reaction mixture was stirred at 25 ° C for 5 min, then warmed to 80 ° C and stirred for 3 h. The reaction mixture was quenched with water (50 mL) and extracted with EtOAc (80 mL × 3), and the combined organic layer was washed with brine (100 mL) and purified by Na 2 SO 4 Drying and concentration gave a residue. The residue was stirred with EtOAc (30 mL), filtered and the filter cake was collected and concentrated to give (Z)-3-(2-chlorobenzylidene)-6-fluoro-4-nitroisobenzofuran-1(3H)-one (1.1 g, 68%) as a yellow solid. MS (ESI): C 15 H 7 ClFNO 4 Calculated mass for m / z, 319.00, observed massless.

[0162] Step 2: (3Z)-3-[(2-chlorophenyl)methylene]-6-fluoro-4-nitro-2-benzofuran-1-one (1.1 g, 3.40 mmol) was added to 4M HCl in MeOH (30 mL). The reaction mixture was stirred at 70 °C for 48 h. The reaction mixture was concentrated to give methyl 2-(2-(2-chlorophenyl)acetyl)-5-fluoro-3-nitrobenzoate (920 mg, 76%) as a yellow solid. MS (ESI): C 16 H 11 ClFNO 5Calculated mass, 351.03, observed m / z 352.0 [M+H] + .

[0163] Step 3: In N 2 To a solution of methyl 2-[2-(2-chlorophenyl)acetyl]-5-fluoro-3-nitrobenzoate (400 mg, 1.14 mmol) and 1-methylpiperidine-4-carbaldehyde (289 mg, 2.27 mmol) in THF (18 mL) and MeOH (3 mL) was added TiCl at 0 °C. 3 HCl solution (5.25 g, 6.82 mmol). The reaction mixture was stirred at 40 °C for 16 h. The reaction mixture was quenched with water (20 mL) and extracted with EtOAc (30 mL×3). The combined organic layers were washed with brine (30 mL) and purified by Na 2 SO 4 Drying and concentration gave a residue. The residue was purified by preparative TLC (MeOH / DCM=1 / 10, rf=0.4), and the fractions with the MS signal of the desired product were collected and concentrated to give 3-(2-chlorophenyl)-7-fluoro-2-(1-methylpiperidin-4-yl)-4-oxo-1,2,3,4-tetrahydroquinoline-5-carboxylic acid methyl ester (120 mg, 24%) as a yellow oil. MS (ESI): C 23 H 24 CIF 2 O 3 Calculated mass, 430.15, observed m / z 431.1 [M+H] + .

[0164] Step 4: A solution of methyl 3-(2-chlorophenyl)-7-fluoro-2-(1-methylpiperidin-4-yl)-4-oxo-2,3-dihydro-1H-quinoline-5-carboxylate (100 mg, 0.23 mmol) in MeOH (8 mL) was stirred at 25 °C for 20 min, and N 2 H 4 .H 2 O (436 mg, 6.96 mmol) and stirred at 25 ° C for 16 h. The reaction mixture was concentrated to give a residue. The residue was purified by preparative HPLC (chromatographic column: Xbridge 5u-C18 150×19 mm, 5 μm mobile phase: ACN--H 2O (0.1% formic acid), B (acetonitrile); flow rate: 20 mL / min, wavelength: 214 / 254 nm) to obtain 9-(2-chlorophenyl)-5-fluoro-8-(1-methylpiperidin-4-yl)-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one (50 mg, 52%) as a white solid. MS (ESI): C 22 H 22 CIF 4 Calculated mass of O, 412.15, observed m / z 413.1 [M+Na] + .

[0165] Step 5: 9-(2-chlorophenyl)-5-fluoro-8-(1-methylpiperidin-4-yl)-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one (50 mg, 0.12 mmol) was separated by preparative SFC (Daicel CHIRALPAK IB 30×250 mm, 10 μm mobile phase A / B: CO 2 / MeOH (0.1% EDA)) = 60 / 40; flow rate: 1.5 mL / min, column temperature: 34 degrees) to obtain (8R, 9R)-9-(2-chlorophenyl)-5-fluoro-8-(1-methylpiperidin-4-yl)-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one (15.0 mg, 30%) as a white solid and (8S, 9S)-9-(2-chlorophenyl)-5-fluoro-8-(1-methylpiperidin-4-yl)-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one (8.5 mg, 17%) as a white solid.

[0166] 0116A:MS(ESI):C 22 H 22 CIF 4 Calculated mass of O, 412.15, observed m / z 413.1 [M+Na] + . 1HNMR (400MHz, DMSO-d6) δ12.44(s,1H),7.50(dd,J=8.0,1.2Hz,1H),7.40(d,J=2.4Hz,1H),7.26( td,J=7.6,1.6Hz,1H),7.18(td,J=7.6,1.2Hz,1H),7.01(dd,J=9.2,2.4Hz,1H),6.90(dd,J=11.2, 2.4Hz,1H),6.66(dd,J=7.6,1.2Hz,1H),4.56(d,J=3.6Hz,1H),3.36–3.30(m,1H),2.95-2.80(m,2 H),2.20(s,3H),1.96(t,J=10.4Hz,1H),1.91–1.75(m,2H),1.72–1.50(m,2H),1.45-1.33(m,2H).

[0167] 0116B:MS(ESI):C 22 H 22 CIF 4 Calculated mass of O, 412.15, observed m / z 413.1 [M+Na] + . HNMR(400MHz,DMSO-d6)δ12.42(s,1H),7.49(dd,J=8.0,1.2Hz,1H),7.36(d,J=2.4Hz,1H) ,7.26(td,J=7.6,1.6Hz,1H),7.23–7.11(m,1H),7.00(dd,J=8.8,2.4Hz,1H),6.95-6.87(m ,1H),6.70-6.66(m,1H),4.55(d,J=3.6Hz,1H),3.34–3.32(m,1H),2.84-2.68(m,2H),2.0 8(s,3H),1.72(t,J=10.4Hz,2H),1.67-1.58(m,2H),1.57–1.46(m,1H),1.40–1.26(m,2H).

[0168] Synthesis of 0117-8, 0117A, 0117B, 0117C and 0117D:

[0169]

[0170] Step 1: To a mixture of methyl 2-[2-(2-chlorophenyl)acetyl]-5-fluoro-3-nitrobenzoate (1.0 g, 2.80 mmol) and tert-butyl (3-formylpyrrolidin-1-yl)carboxylate (1.1 g, 5.60 mmol) in THF (24 mL) and MeOH (4 mL) was added TiCl 3 (10.0 g, 84.00 mmol). The reaction mixture was treated with H 2 O (50 mL) and extracted with EA (2×50 ml). The organic layer was washed with brine (50 mL) and washed with Na 2 SO 4 Dry, filter and concentrate the filtrate to give a residue which is purified by reverse phase column (30% A in B; A: CH 3 CN, B: 0.1% TFA aqueous solution) to give 3-(2-chlorophenyl)-7-fluoro-4-oxo-2-(pyrrolidin-3-yl)-2,3-dihydro-1H-quinoline-5-carboxylic acid methyl ester (600 mg, 52%) as a yellow oil. MS (ESI): C 21 H 20 CIF 2 O 3 Calculated mass, 402.11, found m / z 403.0 [M+H] + .

[0171] Step 2: To a solution of methyl 3-(2-chlorophenyl)-7-fluoro-4-oxo-2-(pyrrolidin-3-yl)-2,3-dihydro-1H-quinoline-5-carboxylate (600 mg, 1.49 mmol) in ACN (10 mL) were added 1-bromo-2-methoxyethane (207 mg, 1.49 mmol) and K 2 CO 3 (618 mg, 4.47 mmol). The reaction mixture was stirred at 80 °C for 16 h. After cooling to room temperature, the reaction mixture was washed with H 2 O (15 mL) and extracted with EA (2×20 mL). The organic layer was washed with brine (20 mL) and washed with anhydrous Na 2 SO 4 Dry and concentrate in vacuo. The residue was purified by flash chromatography (EA) on a silica gel column to give methyl 3-(2-chlorophenyl)-7-fluoro-2-[1-(2-methoxyethyl)pyrrolidin-3-yl]-4-oxo-2,3-dihydro-1H-quinoline-5-carboxylate (400 mg, 55%) as a yellow oil. MS (ESI): C 24 H 26 CIF 2 O 4Calculated mass, 460.16, m / z observed, 461.1 [M+H] + .

[0172] Step 3: To a solution of methyl 3-(2-chlorophenyl)-7-fluoro-2-[1-(2-methoxyethyl)pyrrolidin-3-yl]-4-oxo-2,3-dihydro-1H-quinoline-5-carboxylate (400 mg, 0.87 mmol) in MeOH (5 mL) was added N 2 H 4 H 2 O (217 mg, 4.34 mmol). The reaction mixture was stirred at 25 °C for 16 h. 2 O (15 mL) and extracted with EA (2×20 mL). The organic layer was washed with brine (20 mL) and washed with anhydrous Na 2 SO 4 The residue was dried and concentrated in vacuo. The residue was purified by preparative HPLC (instrument: Waters Prep-HPLC, column: Xbridge preparative c18 5 μm OBD 19×150 mm, A water (0.1% formic acid), B acetonitrile, 10-20% B in 8 min, maintained at 100% B for 2 min, restored to 5% B with 0.5 min, stopped at 13 min; flow rate: 20 ml / min, wavelength: 214 / 254 nm) to obtain 12-(2-chlorophenyl)-7-fluoro-11-[1-(2-methoxyethyl)pyrrolidin-3-yl]-2,3,10-triazatricyclo[7.3.1.0^{5,13}]tridec-1,5(13),6,8-tetraen-4-one ("0117-8"; 260 mg, 65%) as a yellow solid. MS (ESI): C 23 H 24 CIF 4 O 2 Calculated mass, 442.16, observed m / z 443.1 [M+H] + . 1 H NMR (400 MHz, CDCl 3 )δ9.91(d,J=80.0Hz,1H),8.56(s,0H),7.48-7.40(m,1H),7.32-7.27(m,1H),7.25-7.08(m,2H),6.92-6.79(m,1H),6.73-6. 62(m,1H),4.57-4.43(m,1H),3.88-3.53(m,3H),3.43-3.35(m,3H),3.29-2.97(m,2H),2.96-2.49(m,3H),2.45-1.85(m,4H).

[0173] Step 4: 0117-8 (300 mg, 0.68 mmol) was separated by preparative SFC (device: SFC 80 chromatographic column: Daicel CHIRALPAK OX_3, 3×150 mm, 3 μm, mobile phase: A / B: CO 2 / MeOH (0.1% DEA) = 65 / 35; flow rate: 70 g / min, wavelength: UV 214 nm, temperature: 35 ° C) and (device: SFC 80 chromatographic column: Daicel CHIRALPAK IE_3, 3.0 × 150 mm, 3 μm, mobile phase: A / B: CO 2 / MeOH (0.1% DEA) = 50 / 50; flow rate: 70 g / min, wavelength: UV214 nm, temperature: 35°C) to obtain 0117 (13.6 mg, 18%) as a white solid, 0117B (16.5 mg, 22%) as a white solid, 0117C (26.0 mg, 35%) as a white solid and 0117D (28 mg, 37%) as a yellow solid. MS (ESI): C 23 H 24 CIF 4 O 2 Calculated mass, 442.16, observed m / z 443.1 [M+H] + .

[0174] 0117A: 1 H NMR (400MHz, CD 3 OD)δ7.45-7.36(m,1H),7.22-7.02(m,3H),6.82-6.71(m,1H),6.52(d,J=7.6Hz,1H),4.57(d,J=2.4Hz ,1H),3.55-3.42(m,3H),3.33-3.28(m,3H),2.95-2.61(m,6H),2.54-2.38(m,1H),2.19-1.91(m,2H).

[0175] 0117B: 1 H NMR (400MHz, CD 3OD)δ7.50-7.40(m,1H),7.29-7.17(m,2H),7.14-7.03(m,1H),6.91-6.78(m,1H),6.58-6.48(m,1H),4.64-4.59(m,1H ),3.68-3.54(m,3H),3.42-3.31(m,4H),3.29-3.06(m,5H),2.73-2.59(m,1H),2.35-2.23(m,1H),2.22-2.09(m,1H).

[0176] 0117C: 1 H NMR (400MHz, CD 3 OD)δ7.49-7.37(m,1H),7.26-7.06(m,3H),6.85-6.75(m,1H),6.70-7.58(m,1H),4.51-4.47(m ,1H),3.60-3.48(m,3H),3.34(s,3H),2.99-2.58(m,6H),2.49-2.28(m,1H),1.95-1.76(m,2H).

[0177] 0117D: 1 H NMR (400 MHz, CDCl 3 )δ9.76(s,1H),7.54-7.40(m,1H),7.32-7.28(m,2H),7.26(d,J=2.4Hz,1H),7.13-7 .01(m,1H),6.96-6.90(m,1H),4.48(d,J=10.8Hz,1H),3.91(d,J=10.4Hz,1H),3.69 -3.63(m,2H),3.60-3.49(m,2H),3.43-3.35(m,3H),3.06-3.00(m,1H),2.96-2.90( m,1H),2.84-2.76(m,1H),2.64-2.54(m,1H),2.40-2.24(m,2H),2.12-2.00(m,1H).

[0178] Synthesis of 0120A and 0120B

[0179]

[0180] Step 1: In N 2To a solution of methyl 2-[2-(2-chlorophenyl)acetyl]-5-fluoro-3-nitrobenzoate (1 g, 2.80 mmol) and paraformaldehyde (1.01 g, 11.20 mmol) in THF (6 mL) and MeOH (1 mL) cooled to 0°C was added TiCl 3 HCl solution (7 mL). The reaction mixture was stirred at 40 °C for 2 h. The reaction mixture was quenched with water (20 mL) and extracted with EtOAc (30 mL×3). The combined organic layers were washed with brine (30 mL) and purified by Na 2 SO 4 The residue was dried and concentrated to give a residue. The residue was purified by column chromatography on silica gel (12 g) eluting with petroleum ether (19%) containing EtOAc, and the fractions with the MS signal of the desired product were collected and concentrated to give methyl 3-(2-chlorophenyl)-7-fluoro-4-oxo-1,2,3,4-tetrahydroquinoline-5-carboxylate (120 mg, 13%) as a yellow oil. MS (ESI): C 17 H 13 ClFNO 3 Calculated mass, 333.06, m / z observed, 334.2, 336.2 [M+H] + .

[0181] Step 2: 3-(2-chlorophenyl)-7-fluoro-4-oxo-2,3-dihydro-1H-quinoline-5-carboxylic acid methyl ester (140 mg, 0.42 mmol) and N 2 H 4 .H 2 A mixture of 2-nitropropene / MeOH=1 / 1 (8 mL) was stirred at 25°C for 16 h. The reaction mixture was concentrated to give a residue. The residue was purified by preparative HPLC (chromatographic column: Xbridge 5u-C18 150×19 mm, 5 μm mobile phase A: ACN--H 2 O (0.1% formic acid), B (acetonitrile); flow rate: 20 mL / min, wavelength: 214 / 254 nm) to obtain 9-(2-difluorophenyl)-5-fluoro-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one (50 mg, 38%) as a white solid. MS (ESI): C 16 H 11 CIF 3 Calculated mass of O, 315.06, observed m / z 316.0, 318.0 [M+H] + .

[0182] Step 3: 9-(2-chlorophenyl)-5-fluoro-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one (50 mg, 0.16 mmol) was separated by preparative SFC (Daicel CHIRALPAK IB 30×250 mm, 10 μm mobile phase A / B: CO 2 / MeOH (0.1% EDA)) = 50 / 50; flow rate: 1.5 mL / min, column temperature: 37 degrees) to obtain (R)-9-(2-chlorophenyl)-5-fluoro-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazine-3-one (14.6 mg, 29%) as a white solid and (S)-9-(2-chlorophenyl)-5-fluoro-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazine-3-one (13.2 mg, 26%) as a white solid.

[0183] 0120A:MS(ESI):C 16 H 11 CIF 3 Calculated mass of O, 315.06, observed m / z 316.0, 318.0 [M+H] + . 1 H NMR (400 MHz, CDCl 3 )δ9.92(s,1H),7.48-7.37(m,2H),7.25-7.13(m,2H),6.88(dd,J=7.6,1.6H z, 1H), 6.70 (dd, J = 10.4, 2.4Hz, 1H), 4.81-4.71 (m, 2H), 3.76-3.63 (m, 2H).

[0184] 0120B:MS(ESI):C 16 H 11 CIF 3 Calculated mass of O, 315.06, observed m / z 316.0, 318.0 [M+H] + . 1 H NMR (400 MHz, CDCl 3 )δ10.08(s,1H),7.49-7.34(m,2H),7.25-7.11(m,2H),6.87(dd,J=7.6,1.6 Hz, 1H), 6.70 (dd, J = 10.4, 2.4Hz, 1H), 4.84-4.75 (m, 2H), 3.86-3.51 (m, 2H).

[0185] Synthesis of 0121-4, 0121A, 0121B, 0121C and 0121D:

[0186]

[0187] Step 1: To a solution of methyl 2-[2-(2-chlorophenyl)acetyl]-5-fluoro-3-nitrobenzoate (950 mg, 2.70 mmol) in THF (10 mL) and MeOH (2 mL) was added TiCl under ice bath. 3 (12.8 g, 16.20 mmol). The reaction mixture was stirred at 40 °C for 2 h. Then oxacyclopentane-3-carboxaldehyde (811 mg, 8.10 mmol) was added under ice bath. The mixture was stirred at 25 °C for 14 h. The reaction mixture was washed with H 2 O (50 mL) and extracted with EA (2×50 ml). The organic layer was washed with brine (50 mL) and purified by Na 2 SO 4 Dry, filter, and concentrate the filtrate to obtain a residue, which is chromatographed on a silica gel column using PE:EA (3:1) to obtain a yellow solid product (250 mg). MS (ESI): C 21 H 19 ClFNO 4 Calculated mass, 403.10, observed m / z 404.1 [M+H] + .

[0188] Step 2: To a solution of methyl 3-(2-chlorophenyl)-7-fluoro-4-oxo-2-(oxolan-3-yl)-2,3-dihydro-1H-quinoline-5-carboxylate (250 mg, 0.6191 mmol) in MeOH (5 mL) was added NH 2 NH 2 .H 2 O (1.56 g, 24.76 mmol). The reaction mixture was stirred at 25 °C for 3 h. 2 O (15 mL) and extracted with EA (2×20 mL). The organic layer was washed with brine (20 mL) and washed with anhydrous Na 2 SO 4Drying, vacuum concentration. Purify the residue by silica gel column chromatography with PE / EA (2:1) elution to obtain the product (170 mg). Then take 20 mg of the product and purify it by preparative HPLC (instrument: Waters MS triggered preparative LC with QDA detector, column: Xbridge5u C18 150×19 mm, A water (0.1% formic acid), B acetonitrile, 25-55% B in 8 min, keep at 100% B for 2 min, restore to 25% B with 0.5 min, stop at 13 min; flow rate: 20 mL / min, wavelength: 214 / 254 nm), and obtain the target compound ("0121-4"; 2.5 mg, 12%) as a white solid.

[0189] 0121-4:MS(ESI):C 20 H 17 CIF 3 O 2 Calculated mass, 385.10, m / z observed, 386.0 [M+H] + . 1 HNMR(400MHz,MeOD)δ7.48-7.42(m,1H),7.26-7.05(m,3H),6.90-6.75(m,1H),6.64-6.50(m,1H),4.70-4. 45(m,1H),4.01-3.81(m,2H),3.78-3.64(m,2H),3.57-3.46(m,1H),2.58-2.43(m,1H),2.25-1.83(m,2H).

[0190] Step 3: The product was separated by preparative SFC (apparatus: SFC 80, column: Daicel CHIRALCEL IC, 250 mm 30 mm ID, 10 μm, mobile phase: CO 2 / MeOH [0.2% NH 3 (7M MeOH solution)] = 60 / 40; flow rate: 70 g / min, wavelength: UV 214 nm, temperature: 35°C), which produced 35.8 mg of a white solid product (0121), 31.5 mg of a white solid product (0121B), 19.9 mg of a white solid product (0121C), and 26.3 mg of a white solid product (0121D).

[0191] 0121A:MS(ESI):C 20 H 17 CIF 3 O 2 Calculated mass, 385.10, m / z observed, 386.0 [M+H]+ . 1 HNMR(400MHz,MeOD)δ7.37-7.33(m,1H),7.15-6.98(m,3H),6.78-6.71(m,1H),6.54-6.48(m,1H),4.38(d,J=3.2Hz,1 H),3.87-3.72(m,3H),3.61-3.53(m,1H),3.46-3.40(m,1H),2.46-2.33(m,1H),1.93-1.83(m,1H),1.82-1.70(m,1H).

[0192] 0121B:MS(ESI):C 20 H 17 CIF 3 O 2 Calculated mass, 385.10, m / z observed, 386.0 [M+H] + . 1 HNMR(400MHz,MeOD)δ7.38-7.33(m,1H),7.15-6.98(m,3H),6.78-6.71(m,1H),6.54-6.48(m,1H),4.38(d,J=3.2Hz,1 H),3.87-3.72(m,3H),3.61-3.53(m,1H),3.46-3.40(m,1H),2.46-2.33(m,1H),1.94-1.83(m,1H),1.82-1.71(m,1H).

[0193] 0121C:MS(ESI):C 20 H 17 CIF 3 O 2 Calculated mass, 385.10, m / z observed, 386.0 [M+H] + . 1 HNMR(400MHz,MeOD)δ7.42-7.30(m,1H),7.16-6.94(m,3H),6.74-6.66(m,1H),6.49-6.39(m,1H),4.57(d,J=2.0Hz,1 H),3.92-3.82(m,1H),3.71-3.54(m,3H),3.46-3.36(m,1H),2.47-2.32(m,1H),2.15-2.01(m,1H),2.00-1.86(m,1H).

[0194] 0121D:MS(ESI):C 20 H 17 CIF3 O 2 Calculated mass, 385.10, m / z observed, 386.0 [M+H] + . 1 HNMR(400MHz,MeOD)δ7.39-7.33(m,1H),7.16-6.95(m,3H),6.75-6.65(m,1H),6.49-6.39(m,1H),4.57(d,J=2.0Hz,1 H),3.92-3.82(m,1H),3.71-3.54(m,3H),3.46-3.36(m,1H),2.47-2.32(m,1H),2.15-2.01(m,1H),2.00-1.86(m,1H).

[0195] Synthesis of 0171:

[0196]

[0197] To a solution of methyl 3-(2-chlorophenyl)-7-fluoro-4-oxo-2-(pyrrolidin-3-yl)-2,3-dihydro-1H-quinoline-5-carboxylate (20 mg, 0.05 mmol) in MeOH (1 mL) was added N 2 H 4 H 2 O (25mg, 0.50mmol). The reaction mixture was stirred at 25°C for 16 hours. The mixture was concentrated and the residue was purified by preparative HPLC (instrument: Waters Prep-HPLC, column: Xbridge preparative c18 5μm OBD 19×150mm, A water (0.1% formic acid), B acetonitrile, 10-20% B in 8min, kept at 100% B for 2min, restored to 5% B with 0.5min, stopped at 13min; flow rate: 20ml / min, wavelength: 214 / 254nm, 12-(2-chlorophenyl)-7-fluoro-11-(pyrrolidin-3-yl)-2,3,10-triazatricyclo[7.3.1.0^{5,13}]13-1,5(13),6,8-tetraene-4-one (3mg, 15%) was obtained as a yellow solid. MS (ESI): C 20 H 18 CIF 4 Calculated mass of O, 384.12, found m / z 385.0 [M+H] + . 1 H NMR (400MHz, CD 3OD)δ7.55-7.40(m,1H),7.32-7.16(m,2H),7.16-7.11(m,1H),6.93-6.80(m,1H),6.60-6.52(m,1H ),4.68-4.50(m,1H),3.70-3.33(m,3H),3.29-3.08(m,2H),2.74-2.55(m,1H),2.43-1.77(m,2H).

[0198] Synthesis of 0172A and 0172B

[0199]

[0200] Step 1: In N 2 To a solution of methyl 2-[2-(2-chlorophenyl)acetyl]-5-fluoro-3-nitrobenzoate (0.2 g, 0.6 mmol) and acetaldehyde (0.11 g, 2.40 mmol) in THF (4 mL) and MeOH (0.6 mL) was added TiCl at 0 °C. 3 HCl solution (8 mL). The reaction mixture was stirred at 40 °C for 1 h. The reaction mixture was quenched with water (20 mL) and extracted with EtOAc (30 mL×3). The combined organic layers were washed with brine (30 mL) and purified by Na 2 SO 4 The residue was dried and concentrated to give a residue. The residue was purified by column chromatography on silica gel (12 g), eluted with a petroleum ether solution (28%) containing EtOAc, and the fractions with the MS signal of the desired product were collected and concentrated to give 3-(2-chlorophenyl)-7-fluoro-2-methyl-4-oxo-1,2,3,4-tetrahydroquinoline-5-carboxylic acid methyl ester (90 mg, 95%) as a colorless oil. MS (ESI): C 18 H 15 ClFNO 3 Calculated mass, 347.07, m / z observed, 348.2, 350.2 [M+H] + .

[0201] Step 2: A solution of methyl 3-(2-chlorophenyl)-7-fluoro-2-methyl-4-oxo-2,3-dihydro-1H-quinoline-5-carboxylate (100 mg, 0.29 mmol) in MeOH (8 mL) was stirred at 25° C. for 20 min. N 2 H 4 .H 2O (540 mg, 8.63 mmol) and the mixture was stirred at 25 ° C for 16 min. The reaction mixture was directly concentrated to give a residue. The residue was purified by preparative HPLC (chromatographic column: Xbridge 5u-C18 150×19 mm, 5 μm mobile phase A: ACN--H 2 O (0.1% formic acid), B (acetonitrile); flow rate: 20 mL / min, wavelength: 214 / 254 nm) to obtain 9-(2-difluorophenyl)-5-fluoro-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one (50 mg, 38%) as a white solid. MS (ESI): C 17 H 13 CIF 3 Mass calculated for O, 329.07, m / z observed, 330.1, 332.1 [M+H] + .

[0202] Step 3: 9-(2-chlorophenyl)-5-fluoro-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one (90 mg, 0.27 mmol) was separated by preparative SFC (Daicel CHIRALPAK IB 30×250 mm, 10 μm mobile phase A / B: CO 2 / MeOH(0.1%EDA))=65 / 35; flow rate: 2.0mL / min, column temperature: 37 degrees) to obtain (8R,9R)-9-(2-chlorophenyl)-5-fluoro-8-methyl-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one (27.4mg, 30%) as a white solid and (8S,9S)-9-(2-chlorophenyl)-5-fluoro-8-methyl-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one (60.4mg, 67%) as a white solid.

[0203] 0172A and 0172B: MS (ESI): C 17 H 13 CIF 3 Mass calculated for O, 329.07, m / z observed, 330.0, 332.0 [M+H] + . 1 H NMR (400 MHz, CDCl 3)δ9.74(s,1H),7.47-7.45(m,1H),7.38 -7.35(m,1H),7.26-7.22(m,2H),7.01-6.99(m,1H),6.70-6.67(m,1H),4.47-4.45(m,1H),3.92-3.86(m,1H),1.26(d,J=6.4,3H). 1 H NMR (400 MHz, CDCl 3 )δ9.74(s,1H),7.47-7.45(m,1H),7.37-7.33(m,1H),7.26-7.22(m,2H),7.01-6.99( m,1H),6.70-6.67(m,1H),4.47-4.45(m,1H),3.92-3.86(m,1H),1.26(d,J=6.4,3H).

[0204] Synthesis of 0173:

[0205]

[0206] To a solution of methyl 3-(2,4-difluorophenyl)-7-fluoro-4-oxo-2-(pyrrolidin-3-yl)-2,3-dihydro-1H-quinoline-5-carboxylate (40 mg, 0.10 mmol) in MeOH (1 mL) was added N 2 H 4. H 2 O (50mg, 0.99mmol). The reaction mixture was stirred at 25 ° C for 16 hours. The mixture was concentrated and the residue was purified by preparative HPLC (instrument: Waters Prep-HPLC, column: Xbridge preparative c18 5μm OBD 19×150mm, A water (0.1% formic acid), B acetonitrile, 10-20% B in 8min, kept at 100% B for 2min, restored to 5% B with 0.5min, stopped at 13min; flow rate: 20ml / min, wavelength: 214 / 254nm, 12-(2,4-difluorophenyl)-7-fluoro-11-(pyrrolidin-3-yl)-2,3,10-triazatricyclo[7.3.1.0^{5,13}]13-1,5(13),6,8-tetraene-4-one (3mg, 8%) was obtained as a white solid. MS (ESI): C 20 H 17 F 3 N 4 Calculated mass of O, 386.14, found m / z 387.0 [M+H] + . 1 H NMR (400MHz, CD3 OD)δ7.10(d,J=9.2Hz,1H),6.93(s,1H),6.76(s,2H),6.67-6.53(m,1H),4.25(d,J=3 .2Hz,1H),3.52(s,3H),3.05(d,J=10.4Hz,2H),2.59-2.42(m,1H),2.29-1.73(m,2H).

[0207] Synthesis of 0174A and 0174B

[0208]

[0209] Step 1: In N 2 To a solution of methyl 2-[2-(2,4-difluorophenyl)acetyl]-5-fluoro-3-nitrobenzoate (400 mg, 1.13 mmol) in THF / MeOH=5 / 1 (6 mL) was added acetaldehyde (200 mg, 4.53 mmol) and TiCl at 0°C. 3 HCl solution (4 mL). The reaction mixture was stirred at 40 °C for 16 h. The reaction mixture was quenched with water (20 mL) and extracted with EtOAc (30 mL×3). The combined organic layers were washed with brine (30 mL) and purified by Na 2 SO 4 The residue was dried and concentrated to give a residue. The residue was purified by column chromatography on silica gel (12 g) eluting with petroleum ether (29%) containing EtOAc, and the fractions with the MS signal of the desired product were collected and concentrated to give methyl 3-(2,4-difluorophenyl)-7-fluoro-2-methyl-4-oxo-1,2,3,4-tetrahydroquinoline-5-carboxylate (90 mg, 23%) as a yellow oil. MS (ESI): C 18 H 14 F 3 NO 3 Calculated mass, 349.09, observed m / z 350.1 [M+H] + .

[0210] Step 2: To a mixture of methyl 3-(2,4-difluorophenyl)-7-fluoro-2-methyl-4-oxo-2,3-dihydro-1H-quinoline-5-carboxylate (90 mg, 0.26 mmol) in MeOH (4 mL) was added N 2 H 4. H 2 O (4 mL). The reaction mixture was stirred at 25 °C for 3 h. The reaction mixture was concentrated to obtain a residue. The residue was purified by preparative HPLC (chromatographic column: Xbridge5u-C18 150×19 mm, 5 μm mobile phase A: ACN--H2 O (0.1% formic acid), B (acetonitrile); flow rate: 20 mL / min, wavelength: 214 / 254 nm, to obtain 9-(2,4-difluorophenyl)-5-fluoro-8-methyl-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one (45 mg, 53%) as a white solid. MS (ESI): C 17 H 12 F 3 N 3 Mass calculated for O: 331.19, m / z, found: 332.1 [M+H] + .

[0211] Step 3: 9-(2,4-difluorophenyl)-5-fluoro-8-methyl-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one (45 mg, 0.14 mmol) was purified by preparative SFC (Daicel CHIRALPAK AD_3 3×150 mm, 3 μm mobile phase A / B: CO 2 / MeOH (0.1% EDA)) = 75 / 25; flow rate: 2.0 mL / min, column temperature: 37 degrees) to obtain white solid (8R, 9R)-9-(2,4-difluorophenyl)-5-fluoro-8-methyl-2,7,8,9-tetrahydro-3H-pyrido [4,3,2-de] phthalazin-3-one (0174A, 7.9 mg, 18%) and white solid (8S, 9S)-9-(2,4-difluorophenyl)-5-fluoro-8-methyl-2,7,8,9-tetrahydro-3H-pyrido [4,3,2-de] phthalazin-3-one (0174B, 7.8 mg, 17%). MS (ESI): C 17 H 12 F 3 N 3 Mass calculated for O: 331.19, m / z, found: 332.1 [M+H] + .

[0212] 0174A: 1 H NMR (400MHz, CDOD) δ7.24-7.09(m,2H),7.02-6.89(m,2H),6.84-6.79(m,1H),4.09-4.05(m,1H),3.81-3.71(m,1H),1.20(d,J=6.4Hz,3H).

[0213] 0174B: 1H NMR (400MHz, CDOD) δ7.24-7.09(m,2H),7.01-6.91(m,2H),6.83-6.78(m,1H),4.09-4.04(m,1H),3.81-3.71(m,1H),1.19(d,J=6.4Hz,3H).

[0214] Synthesis of 0186A and 0186B:

[0215]

[0216] Step 1: To a solution of 6-fluoro-4-nitro-3H-2-benzofuran-1-one (3.0 g, 15.2 mmol) in THF (30 mL) were added 2-fluoro-4-formylbenzonitrile (2.7 g, 18.2 mmol), TEA (9.2 g, 91.2 mmol), Ac 2 O (23.3 g, 228.0 mmol). The reaction mixture was stirred at 80 °C for 3 h. 2 O (100 mL) to quench the reaction, resulting in a solid precipitate. The mixture was filtered and the solid was washed with EtOH (15 mL×2) and EA (15 mL×2) to obtain a yellow solid product (3.0 g, 54%). MS (ESI): C 16 H 6 F 2 N 2 O 4 Calculated mass 328.03, observed m / z 329.0 [M+H] + .

[0217] Step 2: To a solution of 2-fluoro-4-{[(1Z)-5-fluoro-7-nitro-3-oxo-2-benzofuran-1-ylidene]methyl}benzonitrile (2.5 g, 7.6 mmol) in DCM (50 mL) and THF (50 mL) was added HCl / MeOH (4 M, 19 mL, 76.0 mmol). The reaction mixture was stirred at 70 °C for 16 h. The reaction mixture was concentrated under reduced pressure to give the product as a yellow solid (2.5 g, 82%). MS (ESI): C 17 H 10 F 2 N 2 O 5 Calculated mass 360.06, observed m / z 361.0 [M+H] + .

[0218] Step 3: To a solution of methyl 2-[2-(4-cyano-3-fluorophenyl)acetyl]-5-fluoro-3-nitrobenzoate (950 mg, 2.64 mmol) in THF (20 mL) and MeOH (4 mL) were added HCHO (720 mg, 7.91 mmol), TiCl 3 (12.5 g, 15.82 mmol). The reaction mixture was stirred at 40 °C for 3 h. 2 O (50 mL) and extracted with EA (50 ml x 2). The organic layer was washed with brine (50 mL) and purified by Na 2 SO 4 The filtrate was concentrated to give a residue, which was purified by silica gel column chromatography eluting with PE:EA (4:1) to give the product as a yellow solid (300 mg, 32%). MS (ESI): C 18 H 12 F 2 N 2 O 3 Calculated mass 342.08, observed m / z 343.0 [M+H] + .

[0219] Step 4: To a solution of 3-(4-cyano-3-fluorophenyl)-7-fluoro-4-oxo-2,3-dihydro-1H-quinoline-5-carboxylic acid methyl ester (300 mg, 0.88 mmol) in MeOH (5 mL) was added 85% hydrazine hydroxide solution (1.1 g, 17.53 mmol). The mixture was stirred at 20 ° C for 1 h. The reaction mixture was concentrated under reduced pressure to give a residue, which was purified by flash column with PE: EA (4: 1) to give the product (150 mg, 52%). The product was separated by device: SFC 80, column: Daicel CHIRALCELIE, 250 mm, 30 mm, 10 μm, mobile phase: CO 2 / MeOH[0.2% NH 3 (7 M MeOH solution)] = 80 / 20; flow rate: 2.0 g / min, wavelength: UV 214 nm, temperature: 35°C) to obtain 45.3 mg (0186) of the product as a white solid and 22.2 mg (0186B) as a white solid.

[0220] 0186A:MS(ESI):C 17 H 10 F 2 N 4 Calculated mass of O, 324.08, found m / z 325.0 [M+H] + . 1 H NMR (400 MHz, DMSO-d6 )δ12.36(s,1H),7.90-7.83(m,1H),7.49(dd,J=10.8,1.2Hz,1H),7.41(s,1H),7.26(dd,J=8.0,1.2Hz,1 H), 7.02 (dd, J = 9.2, 2.4Hz, 1H), 6.84 (dd, J = 11.2, 2.4Hz, 1H), 4.37 (t, J = 6.4Hz, 1H), 3.68-3.60 (m, 2H).

[0221] 0186B:MS(ESI):C 17 H 10 F 2 N 4 Calculated mass of O, 324.08, found m / z 325.0 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 )δ12.36(s,1H),7.91-7.83(m,1H),7.49(dd,J=10.8,1.2Hz,1H),7.41(s,1H),7.26(dd,J=8.0,1.2Hz,1 H), 7.02 (dd, J=9.2, 2.4Hz, 1H), 6.84 (dd, J=11.2, 2.4Hz, 1H), 4.36 (d, J=6.0Hz, 1H), 3.67-3.59 (m, 2H).

[0222] Synthesis of 0187A and 0187B:

[0223]

[0224] Step 1: To a solution of 6-fluoro-4-nitro-3H-2-benzofuran-1-one (5.0 g, 25.4 mmol) in THF (50 mL) were added 3-fluoro-4-formylbenzonitrile (4.6 g, 30.4 mmol), TEA (15.4 g, 152.3 mmol) and Ac 2 O (38.9 g, 381.0 mmol). The reaction mixture was stirred at 80 °C for 3 h. 2 O (200 mL) was added to quench the reaction mixture, and a solid precipitated. The solid was then filtered and washed with EtOH (25 mL × 2) and EA (25 mL × 2). The solid was dried under vacuum to obtain a yellow solid product (6.0 g, 69%). MS (ESI): C 16 H 6 F 2 N 2 O 4Calculated mass 328.03, observed m / z 329.0 [M+H] + .

[0225] Step 2: To a solution of 3-fluoro-4-{[(1Z)-5-fluoro-7-nitro-3-oxo-2-benzofuran-1-ylidene]methyl}benzonitrile (6.0 g, 18.3 mmol) in DCM (20 mL) and THF (50 mL) was added HCl / MeOH (4 M, 91.5 mL, 366.0 mmol). The reaction mixture was stirred at 70 °C for 16 h. The reaction mixture was concentrated under reduced vacuum to give the product as a yellow solid (4.5 g, 61%).

[0226] Step 3: To a solution of methyl 2-[2-(4-cyano-2-fluorophenyl)acetyl]-5-fluoro-3-nitrobenzoate (1000 mg, 2.78 mmol) in THF (20 mL) and MeOH (4 mL) were added HCHO (250 mg, 8.33 mmol), TiCl 3 (13.2 g, 16.65 mmol). The reaction mixture was stirred at 40 °C for 3 h. 2 O (20 mL) and extracted with EA (20 mL×2). The organic layer was washed with brine (20 mL) and purified by Na 2 SO 4 The filtrate was concentrated to give a residue, which was purified by silica gel column chromatography eluting with PE:EA (4:1) to give the product as a yellow solid (300 mg, 31%).

[0227] Step 4: To a solution of methyl 3-(4-cyano-2-fluorophenyl)-7-fluoro-4-oxo-2,3-dihydro-1H-quinoline-5-carboxylate (230 mg, 0.672 mmol) in MeOH (5 mL) was added 85% hydrazine hydroxide solution (791.0 mg, 13.44 mmol). The mixture was stirred at 20 ° C for 1 h. The reaction mixture was concentrated under reduced pressure to give a residue, which was purified by flash column with PE: EA (4: 1) to give the product (200 mg, 91%). The product was separated by device: SFC 80, column: Daicel CHIRALCEL OD, 250 mm, 30 mm, 10 μm, mobile phase: CO 2 / MeOH[0.2% NH 3 (7 M MeOH solution)] = 80 / 20; flow rate: 2.0 g / min, wavelength: UV 214 nm, temperature: 35°C) to obtain 75.2 mg (0187) and 87.3 mg (0187B) of a white solid product.

[0228] 0187A:MS(ESI):C 17 H 10 F 2 N 4 Calculated mass of O, 324.08, found m / z 325.0 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ12.35(s,1H),7.90(dd,J=10.4,1.6Hz,1H),7.65(dd,J=8.0,1.6Hz,1H),7.44(s,1H),7.38(t,J =7.6Hz, 1H), 7.03 (dd, J = 9.2, 2.4Hz, 1H), 6.86 (dd, J = 11.6, 2.4Hz, 1H), 4.51 (dd, J = 8.8, 4.8Hz, 1H), 3.71-3.53 (m, 2H).

[0229] 0187B:MS(ESI):C 17 H 10 F 2 N 4 Calculated mass of O, 324.08, found m / z 325.0 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ12.35(s,1H),7.89(dd,J=10.4,1.2Hz,1H),7.65(dd,J=8.0,1.2Hz,1H),7.44(s,1H),7.38(t,J=7.6Hz,1 H), 7.03 (dd, J = 9.2, 2.4Hz, 1H), 6.86 (dd, J = 11.2, 2.4Hz, 1H), 4.51 (dd, J = 8.8, 4.8Hz, 1H), 3.69-3.63 (m, 1H), 3.61-3.53 (m, 1H).

[0230] Synthesis of 0188-5, 0188A and 0188B

[0231]

[0232] Step 1: In N 2To a solution of methyl 2-[2-(4-chloro-3-fluorophenyl)acetyl]-5-fluoro-3-nitrobenzoate (2 g, 5.4 mmol) and paraformaldehyde (490 mg, 16.2 mmol) in THF (20 mL) and MeOH (4 mL) was added a solution of TiCl3 in HCl (24 mL). The reaction mixture was stirred at 40 ° C for 3 h. The reaction mixture was quenched with water (20 mL) and extracted with EtOAc (30 mL×3). The combined organic layer was washed with brine (30 mL) and purified by Na 2 SO 4 The residue was dried and concentrated to give a residue. The residue was purified by column chromatography on silica gel (25 g) eluting with petroleum ether (25%) containing EtOAc, and the fractions with the MS signal of the desired product were collected and concentrated to give methyl 3-(4-chloro-3-fluorophenyl)-7-fluoro-4-oxo-1,2,3,4-tetrahydroquinoline-5-carboxylate (420 mg, 22%) as a yellow oil. MS (ESI): C 17 H 12 CIF 2 NO 3 Calculated mass, 351.05, m / z observed, 352.0 [M+H] + .

[0233] Step 2: In N 2 To a solution of methyl 3-(4-chloro-3-fluorophenyl)-7-fluoro-4-oxo-2,3-dihydro-1H-quinoline-5-carboxylate (420 mg, 1.19 mmol) in MeOH (15 mL) was added N 2 H 4 .H 2 O (598 mg, 11.94 mmol). The reaction mixture was stirred at 25 ° C for 16 h. The reaction mixture was directly concentrated to obtain a residue. The residue was purified by preparative HPLC (chromatographic column: Xbridge 5u-C18 150×19 mm, 5 μm mobile phase A: ACN--H 2 O, B (acetonitrile; flow rate: 20 mL / min, wavelength: 214 / 254 nm) to obtain 9-(4-chloro-3-fluorophenyl)-5-fluoro-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one ("0188-5"; 200 mg, 50%) as a yellow solid. MS (ESI): C 16 H 10 CIF 2 N 3 Calculated mass of O, 333.05, found m / z 334.0 [M+H] + . 1H NMR(400MHz,MeOD)δ7.37(t,J=8.0Hz,1H),7.37(td,J=8.0,4.0Hz,1H),7.03-6 .98(m,1H),6.80(dd,J=10.4,2.4Hz,1H),4.30–4.15(m,1H),3.72-3.60(m,2H).

[0234] Step 3: 9-(4-chloro-3-fluorophenyl)-5-fluoro-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one ("0188-5"; 300 mg, 0.72 mmol) was separated by preparative SFC (Daicel CHIRALPAK AD_3 3×150 mm, 3 μm mobile phase A / B: CO 2 / MeOH (0.1% EDA) = 70 / 30; flow rate: 2.0 mL / min, column temperature: 37 degrees) to obtain (R)-9-(4-chloro-3-fluorophenyl)-5-fluoro-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one (0188A, 87.8 mg, 29%) as a white solid and (S)-9-(4-chloro-3-fluorophenyl)-5-fluoro-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one (0188B, 8778 mg, 26%) as a white solid.

[0235] 0188A:MS(ESI):C 16 H 10 CIF 2 N 3 Calculated mass of O, 333.05, found m / z 334.0 [M+H] + . 1 HNMR (400MHz, MeOD) δ7.37(t,J=8.0Hz,1H),7.18-7.09(m,2H),7.03-6.97(m,1H),6.83-6.74(m,1H),4.28-4.18(m,1H),3.73-3.58(m,2H).

[0236] 0188B:MS(ESI):C 16 H 10 CIF 2 N 3 Calculated mass of O, 333.05, found m / z 334.0 [M+H] + . 1HNMR (400MHz, MeOD) δ7.37(t,J=8.0Hz,1H),7.20-7.06(m,2H),7.03-6.97(m,1H),6.92-6.78(m,1H),4.30-4.16(m,1H),3.72-3.59(m,2H).

[0237] Synthesis of 0189A and 0189B:

[0238]

[0239] Step 1: To a solution of 6-fluoro-4-nitro-3H-2-benzofuran-1-one (10.0 g, 50.7 mmol) in THF (100 mL) were added 4-chloro-2-fluorobenzaldehyde (9.7 g, 60.8 mmol), TEA (30.8 g, 30.4 mmol), Ac 2 O (62.1 g, 60.8 mmol). The reaction mixture was stirred at 80 °C for 3 h. 2 O (500 mL) to quench the reaction mixture. After filtration, the solid was collected and washed with EtOH (2×40 mL) and EA (2×40 mL). The solid was dried in vacuo to give the product as a yellow solid (9.0 g). MS (ESI): C 15 H 6 CIF 2 NO 4 Calculated mass, 337.00, found m / z 337.9 [M+H] + .

[0240] Step 2: To a solution of (3Z)-3-[(4-chloro-2-fluorophenyl)methylene]-6-fluoro-4-nitro-2-benzofuran-1-one (9.0 g, 26.7 mmol) in THF (150 mL) was added HCl / MeOH (67 mL, 268.0 mmol). The reaction mixture was stirred at 70 °C for 16 h. The reaction mixture was concentrated and then diluted with EA (150 mL), H2O (200 mL), and extracted with EA (2×150 mL). The organic layer was washed with brine (150 mL) and purified by anhydrous Na 2 SO 4 The residue was purified by silica gel column chromatography using PE / EA (8:1) as eluent to give the product as a yellow solid (6.0 g). MS (ESI): C 16 H 10 CIF 2 NO 5 Calculated mass, 369.02, observed m / z 392.0 [M+Na] + .

[0241] Step 3: To a solution of methyl 2-[2-(4-chloro-2-fluorophenyl)acetyl]-5-fluoro-3-nitrobenzoate (370 mg, 1.00 mmol) in THF (10 mL) and MeOH (2 mL) were added aqueous formaldehyde (300 mg, 3.00 mmol), TiCl 3 (4.7 g, 6.0 mmol). The reaction mixture was stirred at 40 °C for 3 h. 2 O (50 mL) and extracted with EA (2×50 mL). The organic layer was washed with brine (50 mL) and washed with anhydrous Na 2 SO 4 The residue was purified by silica gel column chromatography using PE / EA (5:1) as eluent to obtain a yellow solid product (180 mg). MS (ESI): C 17 H 12 CIF 2 NO 3 Calculated mass, 351.05, m / z observed, 352.0 [M+H] + .

[0242] Step 4: To a solution of methyl 3-(4-chloro-2-fluorophenyl)-7-fluoro-4-oxo-2,3-dihydro-1H-quinoline-5-carboxylate (180 mg, 0.51 mmol) in MeOH (5 mL) was added NH 2 NH 2 .H 2 O (647 mg, 10.24 mmol). The reaction mixture was stirred at 25 °C for 3 h. 2 O (20 mL) and extracted with EA (2×20 mL). The organic layer was washed with brine (30 mL) and washed with anhydrous Na 2 SO 4 The residue was purified by silica gel column chromatography using PE / EA (1:1) as eluent to give a crude product (90 mg). The crude product was separated by means of the following apparatus: SFC 150, column: Daicel CHIRALCEL OJ, 250 mm, 30 mm ID, 10 μm, mobile phase: CO 2 / MeOH[0.2% NH 3 (7 M MeOH solution)] = 80 / 20; flow rate: 2.0 g / min, wavelength: UV 214 nm, temperature: 35°C), thereby producing 17.6 mg of a white solid product (0189) and 25.0 mg of a white solid product (0189B).

[0243] 0189A:MS(ESI):C 16 H 10 CIF 2 N 3 Calculated mass of O, 333.05, found m / z 334.0 [M+H] + . 1 HNMR (400MHz, MeOD) δ 7.32-7.06 (m, 3H), 7.02-6.92 (1H), 6.82 (dd, J = 10.8, 2.4Hz, 1H), 4.54-4.44 (m, 1H), 3.72-3.54 (m, 2H).

[0244] 0189B:MS(ESI):C 16 H 10 CIF 2 N 3 Calculated mass of O, 333.05, found m / z 334.0 [M+H] + . 1 HNMR (400MHz, MeOD) δ7.27-7.07(m,3H),7.03-6.95(m,1H),6.82(dd,J=10.8,2.4Hz,1H),4.54-4.43(m,1H),3.71-3.54(m,2H).

[0245] Synthesis of 0193A and 0193B:

[0246]

[0247] Step 1: To a solution of 6-fluoro-4-nitro-3H-2-benzofuran-1-one (3.0 g, 15.2 mmol) in THF (30 mL) were added 2-fluoro-4-formylbenzonitrile (2.7 g, 18.2 mmol), TEA (9.2 g, 91.2 mmol) and Ac 2 O (23.3 g, 228.0 mmol). The reaction mixture was stirred at 80 °C for 3 h. 2 O (100 mL) to quench the reaction and produce a solid precipitate. The mixture was filtered and the solid was washed with EtOH (15 mL×2) and EA (15 mL×2) to obtain a yellow solid product (3.0 g, 54%). MS (ESI): C 16 H 6 F 2 N 2 O 4 Calculated mass 328.03, observed m / z 329.0 [M+H] + .

[0248] Step 2: To a solution of 2-fluoro-4-{[(1Z)-5-fluoro-7-nitro-3-oxo-2-benzofuran-1-ylidene]methyl}benzonitrile (2.5 g, 7.6 mmol) in DCM (50 mL) and THF (50 mL) was added HCl / MeOH (4 M, 19 mL, 76.0 mmol). The reaction mixture was stirred at 70 °C for 16 h. The reaction mixture was concentrated under reduced pressure to give the product as a yellow solid (2.5 g, 82%). MS (ESI): C 17 H 10 F 2 N 2 O 5 Calculated mass 360.06, observed m / z 361.0 [M+H] + .

[0249] Step 3: To a solution of methyl 2-[2-(4-cyano-3-fluorophenyl)acetyl]-5-fluoro-3-nitrobenzoate (800 mg, 2.22 mmol) in THF (20 mL) and MeOH (4 mL) was added CH 3 CHO (294 mg, 6.66 mmol) and TiCl 3 (10.6 g, 13.32 mmol). The reaction mixture was stirred at 40 °C for 3 h. 2 O (50 mL) and extracted with EA (50 ml x 2). The organic layer was washed with brine (50 mL) and purified by Na 2 SO 4 The filtrate was concentrated to give a residue, which was purified by silica gel column chromatography eluting with PE:EA (4:1) to give the product as a yellow solid (320 mg, 38%). MS (ESI): C 19 H 14 F 2 N 2 O 3 Calculated mass 356.10, observed m / z 357.0 [M+H] + .

[0250] Step 4: To a solution of 3-(4-cyano-3-fluorophenyl)-7-fluoro-2-methyl-4-oxo-2,3-dihydro-1H-quinoline-5-carboxylic acid methyl ester (320 mg, 0.90 mmol) in MeOH (10 mL) was added 85% hydrazine hydroxide solution (1.1 g, 17.96 mmol). The mixture was stirred at 20 ° C for 1 h. The reaction mixture was concentrated under reduced pressure to give a residue, which was purified by flash column with PE: EA (4: 1) to give the product (150 mg, 58%). Then separated by device: SFC 80, column: Daicel CHIRALCEL IE, 250 mm, 30 mm, 10 μm, mobile phase: CO 2 / MeOH [0.2% NH 3 (7 M MeOH solution)] = 80 / 20; flow rate: 2.0 g / min, wavelength: UV 214 nm, temperature: 35°C) to obtain 52.5 mg (0193) and 56.6 mg (0193B) of a white solid product.

[0251] 0193A:MS(ESI):C 18 H 12 F 2 N 4 Calculated mass of O, 338.10, found m / z 339.0 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ12.29(s,1H),7.90(t,J=7.2Hz,1H),7.54(d,J=10.8Hz,1H),7.42(s,1H),7.34(d,J=8.0Hz,1H),7 .00(dd,J=9.2,2.4Hz,1H),6.83(dd,J=11.2,2.4Hz,1H),4.03(d,J=10.0Hz,1H),3.86-3.77(m,1H),1.07(d,J=6.0Hz,3H).

[0252] 0193B:MS(ESI):C 18 H 12 F 2 N 4 Calculated mass of O, 338.10, found m / z 339.0 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ12.29(s,1H),7.90(t,J=7.6Hz,1H),7.54(d,J=10.4Hz,1H),7.42(s,1H),7.34(d,J=8.4Hz,1H),7 .00(dd,J=9.2,2.4Hz,1H),6.83(dd,J=11.2,2.4Hz,1H),4.03(d,J=9.6Hz,1H),3.86-3.77(m,1H),1.07(d,J=6.0Hz,3H).

[0253] Synthesis of 0194A and 0194B:

[0254]

[0255] Step 1: To a solution of 6-fluoro-4-nitro-3H-2-benzofuran-1-one (5.0 g, 25.4 mmol) in THF (50 mL) were added 3-fluoro-4-formylbenzonitrile (4.6 g, 30.4 mmol), TEA (15.4 g, 152.3 mmol) and Ac 2 O (38.9 g, 381.0 mmol). The reaction mixture was stirred at 80 °C for 3 h. 2 O (200 mL) to quench the reaction and produce a solid precipitate. The mixture was filtered and the solid was washed with EtOH (15 mL×2) and EA (15 mL×2) to obtain a yellow solid product (6.0 g, 69%). MS (ESI): C 16 H 6 F 2 N 2 O 4 Calculated mass 328.03, observed m / z 329.0 [M+H] + .

[0256] Step 2: To a solution of 3-fluoro-4-{[(1Z)-5-fluoro-7-nitro-3-oxo-2-benzofuran-1-ylidene]methyl}benzonitrile (6.0 g, 18.3 mmol) in DCM (20 mL) and THF (50 mL) was added HCl / MeOH (91.5 mL, 366.0 mmol). The reaction mixture was stirred at 70 °C for 16 h. The reaction mixture was concentrated under reduced vacuum to give the product as a yellow solid (4.5 g, 61%).

[0257] Step 3: To a solution of methyl 2-[2-(4-cyano-2-fluorophenyl)acetyl]-5-fluoro-3-nitrobenzoate (1000 mg, 2.78 mmol) in THF (20 mL) and MeOH (4 mL) was added CH3 CHO (367 mg, 8.33 mmol), TiCl 3 (13.2 g, 16.65 mmol). The reaction mixture was stirred at 40 °C for 3 h. 2 O (20 mL) and extracted with EA (20 mL×2). The organic layer was washed with brine (20 mL) and purified by Na 2 SO 4 The filtrate was concentrated to give a residue, which was purified by silica gel column chromatography eluting with PE:EA (5:1) to give the product as a yellow solid (600 mg, 60%).

[0258] Step 4: To a solution of 3-(4-cyano-2-fluorophenyl)-7-fluoro-2-methyl-4-oxo-2,3-dihydro-1H-quinoline-5-carboxylic acid methyl ester (400 mg, 1.23 mmol) in MeOH (5 mL) was added 85% hydrazine hydroxide solution (199 mg, 3.37 mmol). The mixture was stirred at 20 ° C for 1 h. The reaction mixture was concentrated under reduced pressure to give a residue, which was purified by flash column with PE: EA (4: 1) to give the product (200 mg, 91%). The product was separated by device: SFC 80, column: Daicel CHIRALCEL OD, 250 mm, 30 mm, 10 μm, mobile phase: CO 2 / MeOH[0.2% NH 3 (7 M MeOH solution)] = 80 / 20; flow rate: 2.0 g / min, wavelength: UV 214 nm, temperature: 35°C) to obtain 42.0 mg (0194A) and 38.9 mg (0194B) of a white solid product.

[0259] 0194A:MS(ESI):C 18 H 12 F 2 N 4 Calculated mass of O, 338.10, found m / z 339.0 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ12.29(s,1H),7.89(dd,J=10.0,1.2Hz,1H),7.71(dd,J=8.0,1.2Hz,1H),7.57(t,J=7.6Hz,1H),7.45(s, 1H), 7.02 (dd, J = 9.2, 2.4Hz, 1H), 6.84 (dd, J = 11.2, 2.4Hz, 1H), 4.17 (d, J = 10.8Hz, 1H), 3.87-3.77 (m, 1H), 1.10 (d, J = 6.0Hz, 3H).

[0260] 0194B:MS(ESI):C 18 H 12 F 2 N 4 Calculated mass of O, 338.10, found m / z 339.0 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ12.29(s,1H),7.89(dd,J=10.4,1.2Hz,1H),7.71(dd,J=8.0,1.2Hz,1H),7.57(t,J=7.6Hz,1H),7.45(s, 1H), 7.02 (dd, J = 9.2, 2.4Hz, 1H), 6.84 (dd, J = 11.2, 2.4Hz, 1H), 4.17 (d, J = 10.8Hz, 1H), 3.86-3.78 (m, 1H), 1.10 (d, J = 6.4Hz, 3H).

[0261] Synthesis of 0195-5, 0195A, and 0195B

[0262]

[0263] Step 1: In N 2 To a solution of methyl 2-(2-(4-chloro-3-fluorophenyl)acetyl)-5-fluoro-3-nitrobenzoate (1 g, 2.7 mmol) and acetaldehyde (0.59 g, 13.5 mmol) in THF (12 mL) and MeOH (2 mL) was added TiCl 3 HCl solution (14 mL). The reaction mixture was stirred at 40 °C for 3 h. The reaction mixture was quenched with water (20 mL) and extracted with EtOAc (30 mL×3). The combined organic layers were washed with brine (30 mL) and purified by Na 2 SO 4The residue was dried and concentrated to give a residue. The residue was purified by column chromatography on silica gel (25 g) eluting with petroleum ether (25%) containing EtOAc, and the fractions with the MS signal of the desired product were collected and concentrated to give 3-(4-chloro-3-fluorophenyl)-7-fluoro-2-methyl-4-oxo-1,2,3,4-tetrahydroquinoline-5-carboxylic acid methyl ester (300 mg, 30%) as a white solid. MS (ESI): C 18 H 14 CIF 2 NO 3 Calculated mass, 365.06, observed m / z 366.1 [M+H] + .

[0264] Step 2: In N 2 To a solution of methyl 3-(4-chloro-3-fluorophenyl)-7-fluoro-2-methyl-4-oxo-2,3-dihydro-1H-quinoline-5-carboxylate (300 mg, 0.82 mmol) in MeOH (15 mL) was added N 2 H 4 .H 2 O (411 mg, 8.2 mmol). The reaction mixture was stirred at 25 °C for 3 h. The reaction mixture was concentrated to obtain a residue. The residue was purified by preparative HPLC (chromatographic column: Xbridge 5u-C18 150×19 mm, 5 μm mobile phase A: ACN--H 2 O, B (acetonitrile; flow rate: 20 mL / min, wavelength: 214 / 254 nm) to obtain 9-(2-chlorophenyl)-5-fluoro-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one ("0195-5"; 200 mg, 72%) as a white solid. MS (ESI): C 17 H 12 CIF 2 N 3 Calculated mass of O, 347.06, observed m / z 348.1 [M+H] + . 1 H NMR(400MHz,MeOD)δ7.43(t,J=8.0Hz,1H),7.19-7.10(m,2H),7.08-7.02(m,1H),6.82 (dd,J=11.2,2.4Hz,1H),3.89-3.85(m,1H),3.78-3.72(m,1H),1.21(d,J=6.4Hz,3H).

[0265] Step 3: 9-(4-chloro-3-fluorophenyl)-5-fluoro-8-methyl-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one (“0195-5”; 250 mg, 0.72 mmol) was separated by preparative SFC (Daicel CHIRALPAK AD_33×150 mm, 3 μm mobile phase A / B: CO 2 / MeOH (0.1% EDA)) = 70 / 30; flow rate: 2.0 mL / min, column temperature: 37 degrees) to obtain off-white solid (8R, 9R)-9-(4-chloro-3-fluorophenyl)-5-fluoro-8-methyl-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one (0195, 100.7 mg, 40%) and off-white solid (8S, 9S)-9-(4-chloro-3-fluorophenyl)-5-fluoro-8-methyl-2,7,8,9-tetrahydro-3H-pyrido[4,3,2-de]phthalazin-3-one (0195B, 85.7 mg, 34%).

[0266] 0195A:MS(ESI):C 17 H 12 CIF 2 N 3 Calculated mass of O, 347.06, observed m / z 348.1 [M+H] + . 1 HNMR(400MHz,MeOD)δ7.41(t,J=8.0Hz,1H),7.16-7.07(m,2H),7.03(dd,J=8.4,1.6Hz,1H) ,6.79(dd,J=11.2,2.4Hz,1H),3.87-3.83(m,1H),3.79-3.70(m,1H),1.19(d,J=6.4Hz,3H).

[0267] 0195B:MS(ESI):C 17 H 12 CIF 2 N 3 Calculated mass of O, 347.06, observed m / z 348.1 [M+H] + . 1 HNMR(400MHz,MeOD)δ7.41(t,J=8.0Hz,1H),7.21-7.07(m,2H),7.03(dd,J=8.4,1.6Hz,1H) ,6.79(dd,J=11.2,2.4Hz,1H),3.88-3.82(m,1H),3.80-3.70(m,1H),1.19(d,J=6.4Hz,3H).

[0268] Synthesis of 0196A and 0196B:

[0269]

[0270] Step 1: To a solution of 6-fluoro-4-nitro-3H-2-benzofuran-1-one (10.0 g, 50.7 mmol) in THF (100 mL) were added 4-chloro-2-fluorobenzaldehyde (9.7 g, 60.8 mmol), TEA (30.8 g, 30.4 mmol), Ac 2 O (62.1 g, 60.8 mmol). The reaction mixture was stirred at 80 °C for 3 h. 2 O (500 mL) to quench the reaction mixture. The solid was collected by filtration and washed with EtOH (2×40 mL), EA (2×40 mL). The solid was dried in vacuo to give the product as a yellow solid (9.0 g). MS (ESI): C 15 H 6 CIF 2 NO 4 Calculated mass, 337.00, found m / z 337.9 [M+H] + .

[0271] Step 2: To a solution of (3Z)-3-[(4-chloro-2-fluorophenyl)methylene]-6-fluoro-4-nitro-2-benzofuran-1-one (9.0 g, 26.7 mmol) in THF (150 mL) was added HCl / MeOH (67 mL, 268.0 mmol). The reaction mixture was stirred at 70 °C for 16 h. The reaction mixture was concentrated and then diluted with EA (150 mL), H2O (200 mL), and extracted with EA (2×150 mL). The organic layer was washed with brine (150 mL) and purified by anhydrous Na 2 SO 4 The residue was purified by silica gel column chromatography using PE / EA (8:1) as eluent to give the product as a yellow solid (6.0 g). MS (ESI): C 16 H 10 CIF 2 NO 5 Calculated mass, 369.02, observed m / z 392.0 [M+Na] + .

[0272] Step 3: To a solution of methyl 2-[2-(4-chloro-2-fluorophenyl)acetyl]-5-fluoro-3-nitrobenzoate (500 mg, 1.35 mmol) in THF (10 mL) and MeOH (2 mL) were added acetaldehyde (72 mg, 1.62 mmol), TiCl3 (6.4 g, 8.11 mmol). The reaction mixture was stirred at 40 °C for 3 h. 2 O (50 mL) and extracted with EA (2×50 mL). The organic layer was washed with brine (50 mL) and washed with anhydrous Na 2 SO 4 The residue was purified by silica gel column chromatography using PE / EA (5:1) as eluent to obtain a yellow solid product (300 mg). MS (ESI): C 18 H 14 CIF 2 NO 3 Calculated mass, 365.06, found m / z 366.0 [M+H] + .

[0273] Step 4: To a solution of methyl 3-(4-chloro-2-fluorophenyl)-7-fluoro-4-oxo-2,3-dihydro-1H-quinoline-5-carboxylate (300 mg, 0.85 mmol) in MeOH (5 mL) was added NH 2 NH 2 .H 2 O (1.1 g, 17.06 mmol). The reaction mixture was stirred at 25 °C for 3 h. 2 O (20 mL) and extracted with EA (2×30 mL). The organic layer was washed with brine (30 mL) and washed with anhydrous Na 2 SO 4 The residue was purified by silica gel column chromatography using PE / EA (1:1) as eluent to give a crude product (150 mg). The crude product was separated by the following apparatus: SFC 150, column: Daicel CHIRALCEL OJ, 250 mm 30 mm ID, 10 μm, mobile phase: CO 2 / MeOH [0.2% NH 3 (7 M MeOH solution)] = 70 / 30; flow rate: 2.0 g / min, wavelength: UV 214 nm, temperature: 35°C), thereby producing 45.9 mg of a white solid product (0196) and 49.5 mg of a white solid product (0196B).

[0274] 0196A:MS(ESI):C 17 H 12 CIF 2 N 3 Calculated mass of O, 347.06, observed m / z 348.0 [M+H] + . 1HNMR (400MHz, MeOD) δ7.32–7.10(m,4H),6.83(dd,J=10.8,2.4Hz,1H),4.10(d,J=9.6Hz,1H),3.85–3.73(m,1H),1.22(d,J=6.4Hz,3H).

[0275] 0196B:MS(ESI):C 17 H 12 CIF 2 N 3 Calculated mass of O, 347.06, observed m / z 348.0 [M+H] + . 1 HNMR (400MHz, MeOD) δ7.30–7.02(m,4H),6.81(dd,J=10.8,2.4Hz,1H),4.08(d,J=9.6Hz,1H),3.86–3.71(m,1H),1.20(d,J=6.4Hz,3H).

[0276] Synthesis of 0206A and 0206B:

[0277]

[0278] Step 1: To a mixture of 6-fluoro-4-nitro-3H-2-benzofuran-1-one (10.0 g, 50.73 mmol) and 2-chlorobenzaldehyde (14.3 g, 101.46 mmol) in THF (500 mL) were added TEA (15.4 g, 152.19 mmol) and Ac 2 O (31.1 g, 304.38 mmol). The mixture was stirred at 80°C for 16 hours. The reaction mixture was filtered. The filter cake was washed with water (200 mL×2) and dried under reduced pressure to obtain (3Z)-3-[(2-chlorophenyl)methylene]-6-fluoro-4-nitro-2-benzofuran-1-one (10.0 g, 56%) as a yellow solid. MS (ESI): C 15 H 7 ClFNO 4 Calculated mass, 319.0, found m / z 319.9 [M+H] + .

[0279] Step 2: A mixture of [methylene]-6-fluoro-4-nitro-2-benzofuran-1-one (10.0 g, 31.28 mmol) in HCl / EA (500 mL, 4 m in EA) was stirred at 75 ° C for 16 hours. The reaction mixture was filtered. The filter cake was washed with MeOH (100 mL×2) and dried under reduced pressure to give methyl 2-[2-(2-chlorophenyl)acetyl]-5-fluoro-3-nitrobenzoate (10.0 g, 82%) as a yellow solid. MS (ESI): C 16 H 11 ClFNO 5 Calculated mass, 351.0, observed m / z 352.0 [M+H] + .

[0280] Step 3: To a solution of methyl 2-[2-(2-chlorophenyl)acetyl]-5-fluoro-3-nitrobenzoate (2.0 g, 5.69 mmol) in MeOH (10 mL) and THF (50 mL) was added TiCl 3 (20.3 g, 34.12 mmol) and propionaldehyde (991 mg, 17.06 mmol). The reaction mixture was stirred at 40 ° C for 6 hours. The mixture was diluted with water (50 mL) and extracted with EA (100 mL×3). The organic layer was concentrated and the residue was purified by flash chromatography on a silica gel column (PE: EA = 3: 1) to give 3-(2-chlorophenyl)-2-ethyl-7-fluoro-4-oxo-2,3-dihydro-1H-quinoline-5-carboxylic acid methyl ester (1.0 g, 44%) as a yellow oil. MS (ESI): C 19 H 17 ClFNO 3 Calculated mass, 361.1, observed m / z 362.1 [M+H] + .

[0281] Step 4: To a mixture of methyl 3-(2-chlorophenyl)-2-ethyl-7-fluoro-4-oxo-2,3-dihydro-1H-quinoline-5-carboxylate (750 mg, 2.07 mmol) in MeOH (20 mL) was added N 2 H 4 .H 2 O (199 mg, 6.22 mmol). The reaction mixture was stirred at 25°C for 16 hours. The mixture was concentrated under reduced pressure to give a residue, which was purified by reverse phase column (40% A in B; A: CH 3CN, B: 0.1% FA aqueous solution) to obtain 12-(2-chlorophenyl)-11-ethyl-7-fluoro-2,3,10-triazatricyclo[7.3.1.0^{5,13}]trideca-1,5(13),6,8-tetraen-4-one (300 mg, 40%) as a yellow solid. MS (ESI): C 18 H 15 CIF 3 Calculated mass of O, 343.1, observed m / z 344.1 [M+H] + .

[0282] Step 5: 12-(2-chlorophenyl)-11-ethyl-7-fluoro-2,3,10-triazatricyclo[7.3.1.0^{5,13}]trideca-1,5(13),6,8-tetraen-4-one (300 mg, 0.87 mmol) was separated by preparative SFC (apparatus: SFC 150 column: Daicel CHIRALCEL IE, 250 mm×30 mm ID mobile phase: CO 2 / MeOH[0.2% NH 3 (7M MeOH solution)] = 65 / 35 flow rate: 80 g / min, wavelength: UV 214 nm, temperature: 35°C) to obtain white solid (11S, 12S)-12-(2-chlorophenyl)-11-ethyl-7-fluoro-2,3,10-triazatricyclo[7.3.1.0^{5,13}]tridec-1,5(13),6,8-tetraen-4-one (57.4 mg, 19%) and white solid (11R, 12R)-12-(2-chlorophenyl)-11-ethyl-7-fluoro-2,3,10-triazatricyclo[7.3.1.0^{5,13}]tridec-1,5(13),6,8-tetraen-4-one (32.4 mg, 11%).

[0283] 0206A:MS(ESI):C 18 H 15 CIF 3 Calculated mass of O, 343.1, observed m / z 344.1 [M+H] + . 1 HNMR(400MHz,DMSO)δ12.34(s,1H),7.52-7.48(m,1H),7.41-7.20(m,3H),7.10-6.85(m, 3H), 4.38 (d, J = 7.2Hz, 1H), 3.71-3.60 (m, 1H), 1.63-1.35 (m, 2H), 0.95 (t, J = 7.2Hz, 3H).

[0284] 0206B:MS(ESI):C18 H 15 CIF 3 Calculated mass of O, 343.1, observed m / z 344.1 [M+H] + . 1 HNMR(400MHz,DMSO)δ12.34(s,1H),7.53-7.47(m,1H),7.35-7.21(m,3H),7.04-6.86(m, 3H), 4.38 (d, J = 7.2Hz, 1H), 3.70-3.61 (m, 1H), 1.64-1.35 (m, 2H), 0.95 (t, J = 7.2Hz, 3H).

[0285] Synthesis of 0207A and 0207B:

[0286]

[0287] Step 1: To a solution of methyl 2-[2-(2-chlorophenyl)acetyl]-5-fluoro-3-nitrobenzoate (2.0 g, 5.69 mmol) in MeOH (10 mL) and THF (50 mL) was added TiCl 3 (20.3 g, 34.12 mmol) and cyclopropanecarboxaldehyde (797 mg, 11.37 mmol). The reaction mixture was stirred at 40 ° C for 6 hours. The mixture was diluted with water (50 mL) and extracted with EA (100 mL×3). The organic layer was concentrated. The residue was flash chromatographed on a silica gel column (PE: EA = 3: 1) to give 3-(2-chlorophenyl)-2-cyclopropyl-7-fluoro-4-oxo-2,3-dihydro-1H-quinoline-5-carboxylic acid methyl ester (1.0 g, 42%) as a yellow oil. MS (ESI): C 20 H 17 ClFNO 3 Calculated mass, 373.1, found m / z 374.0 [M+H] + .

[0288] Step 2: To a mixture of methyl 3-(2-chlorophenyl)-2-cyclopropyl-7-fluoro-4-oxo-2,3-dihydro-1H-quinoline-5-carboxylate (750 mg, 2.01 mmol) in MeOH (20 mL) was added N 2 H 4 .H 2 O (193 mg, 6.02 mmol). The reaction mixture was stirred at 25°C for 16 hours. The mixture was concentrated under reduced pressure to give a residue, which was purified by reverse phase column (40% A in B; A: CH 3CN, B: 0.1% FA aqueous solution) to obtain 12-(2-chlorophenyl)-11-cyclopropyl-7-fluoro-2,3,10-triazatricyclo[7.3.1.0^{5,13}]trideca-1,5(13),6,8-tetraen-4-one (300 mg, 29%) as a yellow solid. MS (ESI): C 20 H 17 ClFNO 3 Calculated mass, 373.1, found m / z 374.0 [M+H] + .

[0289] Step 3: 12-(2-chlorophenyl)-11-cyclopropyl-7-fluoro-2,3,10-triazatricyclo[7.3.1.0^{5,13}]trideca-1,5(13),6,8-tetraen-4-one (300 mg, 0.84 mmol) was separated by preparative SFC (apparatus: SFC 150 column: Daicel CHIRALCEL IE, 250 mm×30 mm ID mobile phase: CO 2 / MeOH[0.2% NH 3 (7M MeOH solution)] = 65 / 35 flow rate: 80 g / min, wavelength: UV 214 nm, temperature: 35°C) to obtain white solid (11S, 12S)-12-(2-chlorophenyl)-11-cyclopropyl-7-fluoro-2,3,10-triazatricyclo[7.3.1.0^{5,13}]trideca-1,5(13),6,8-tetraen-4-one (0207, 10.9 mg, 4%) and white solid (11R, 12R)-12-(2-chlorophenyl)-11-cyclopropyl-7-fluoro-2,3,10-triazatricyclo[7.3.1.0^{5,13}]trideca-1,5(13),6,8-tetraen-4-one (0207B, 41.4 mg, 14%).

[0290] 0207A:MS(ESI):C 18 H 15 CIF 3 Calculated mass of O, 343.1, observed m / z 344.1 [M+H] + . 1 HNMR(400MHz,DMSO)δ12.36(s,1H),7.50-7.45(m,1H),7.42-7.22(m,3H),7.06-6.88(m,3H),4.5 5(d,J=6.8Hz,1H),2.91-2.83(m,1H),1.10-0.94(m,1H),0.55-0.22(m,3H),0.06--0.10(m,1H).

[0291] 0207B:MS(ESI):C 18 H 15 CIF 3 Calculated mass of O, 343.1, observed m / z 344.1 [M+H] + . 1 HNMR(400MHz,DMSO)δ12.37(s,1H),7.52-7.45(m,1H),7.36(s,1H),7.31-7.16(m,2H),7.09-6.78(m,3 H), 4.55 (d, J = 6.8Hz, 1H), 2.95-2.82 (m, 1H), 1.11-0.90 (m, 1H), 0.53-0.26 (m, 3H), 0.04--0.07 (m, 1H).

[0292] Synthesis of 0210A and 0210B:

[0293]

[0294] Step 1: To a solution of methyl 2-[2-(2-chlorophenyl)acetyl]-5-fluoro-3-nitrobenzoate (1.0 g, 2.84 mmol) in MeOH (4 mL) and THF (20 mL) was added oxane-4-carbaldehyde (649 mg, 5.69 mmol) and TiCl 3 (10.1 g, 17.06 mmol). The reaction mixture was stirred at 40 °C for 16 hours. The reaction mixture was concentrated under reduced pressure. The mixture was diluted with water (30 mL) and extracted with EA (50 mL×3). The organic layer was concentrated and the residue was purified by flash chromatography on a silica gel column (PE:EA=3:1) to give 3-(2-chlorophenyl)-7-fluoro-2-(oxacyclohexane-4-yl)-4-oxo-2,3-dihydro-1H-quinoline-5-carboxylic acid methyl ester (800 mg, 61%) as a yellow solid. MS (ESI): C 22 H 21 ClFNO 4 Calculated mass, 417.1, observed m / z 418.1 [M+H] + .

[0295] Step 2: To a mixture of methyl 3-(2-chlorophenyl)-7-fluoro-2-(oxan-4-yl)-4-oxo-2,3-dihydro-1H-quinoline-5-carboxylate (750 mg, 1.79 mmol) in MeOH (10 mL) was added N 2 H 4 .H 2O (173 mg, 5.38 mmol). The reaction mixture was stirred at 60°C for 16 hours. The mixture was concentrated under reduced pressure to give a residue, which was purified by reverse phase column (40% A in B; A: CH 3 CN, B: 0.1% FA aqueous solution) to obtain 12-(2-chlorophenyl)-7-fluoro-11-(oxan-4-yl)-2,3,10-triazatricyclo[7.3.1.0^{5,13}]trideca-1,5(13),6,8-tetraen-4-one (300 mg, 40%) as a yellow solid. MS (ESI): C 21 H 19 CIF 3 O 2 Calculated mass, 399.1, observed m / z 400.1 [M+H] + .

[0296] Step 3: 12-(2-chlorophenyl)-7-fluoro-11-(oxan-4-yl)-2,3,10-triazatricyclo[7.3.1.0^{5,13}]tridec-1,5(13),6,8-tetraen-4-one (140 mg, 0.35 mmol) was separated by preparative SFC (instrument: SFC 150 column: Daicel CHIRALPAK IA 4.6×250 mm, 5 μm. Mobile phase: CO 2 / MeOH [0.2% NH 3 (7M MeOH solution)] = 60 / 40 Flow rate: 80 g / min, Wavelength: UV 214nm, temperature: 35°C) to give (11S, 12S)-12-(2-chlorophenyl)-7-fluoro-11-(oxan-4-yl)-2,3,10-triazatricyclo[7.3.1.0^{5,13}]tridec-1,5(13),6,8-tetraen-4-one (0210, 17.5mg, 13%) and (11R, 12R)-12-(2-chlorophenyl)-7-fluoro-11-(oxan-4-yl)-2,3,10-triazatricyclo[7.3.1.0^{5,13}]tridec-1,5(13),6,8-tetraen-4-one (0210B, 7mg, 5%) as a white solid.

[0297] 0210A:MS(ESI):C 21 H 19 CIF 3 O 2 Calculated mass, 399.1, observed m / z 400.1 [M+H] + . 1 HNMR (400 MHz, DMSO-d 6)δ12.44(s,1H),7.51(d,J=8.0Hz,1H),7.38(d,J=2.8Hz,1H),7.31-7.23(m,1H),7.22-7.15(m,1H),7.01(dd,J=9.2,2.4Hz,1H),6.90(dd, J=11.2,2.4Hz,1H),6.70-6.63(m,1H),4.57(d,J=3.2Hz,1H),3.93-3.76(m,2H),3.25-3.05(m,2H),1.76-1.47(m,4H),1.39-1.25(m,1H).

[0298] 0210B:MS(ESI):C 21 H 19 CIF 3 O 2 Calculated mass, 399.1, observed m / z 400.1 [M+H] + . 1 HNMR (400 MHz, DMSO-d 6 )δ12.45(s,1H),7.51(d,J=8.0Hz,1H),7.40(d,J=2.8Hz,1H),7.30-7.23(m,1H),7.22-7.15(m,1H),7.01(dd,J=9.2,2.4Hz,1H),6.90(dd, J=11.2,2.4Hz,1H),6.69-6.63(m,1H),4.57(d,J=3.2Hz,1H),3.92-3.78(m,2H),3.26-3.06(m,2H),1.75-1.47(m,4H),1.44-1.17(m,1H).

[0299] Additional Exemplary Compounds of the Present Technology

[0300] Additional compounds of the present technology were synthesized and isolated according to similar methods and procedures as described above, wherein such additional compounds include (but are not limited to):

[0301]

[0302]

[0303]

[0304]

[0305]

[0306]

[0307]

[0308]

[0309] PARP mass spectrometry protocol

[0310] Materials and Reagents: PARP1 enzyme was purchased from BPS Bioscience (Cat. No. 80501). Tris-HCl (pH 8.0) was purchased from Corning (Cat. No. 46-031-CM). Magnesium chloride was purchased from Thermo Fisher Scientific (formerly HoneywellFluka, Cat. No. 63020-1L). All other assay components, activated DNA (Cat. No. D4522), core histones (Cat. No. SRP6590), β-nicotinamide adenine dinucleotide (β-NAD – Cat. No. N8285), 3ABA PARP1 small molecule inhibitor (from PARP1 enzyme activity assay kit, Cat. No. 17-10149), sodium chloride (NaCl, Cat. No. S6546-1L), Triton X-100 (Cat. No. 93443), dithiothreitol (DTT, Cat. No. 43816-250ML) were purchased from Millipore Sigma.

[0311] Assay Buffer: The assay buffer contained the following reagents: 50 mM Tris-HCl pH 8.0, 50 mM NaCl, 10 mM MgCl2, 0.01% Triton X-100 and 1 mM DTT.

[0312] program:

[0313] The PARP1 enzyme assay was performed in a 384-well plate in a total volume of 20 μL in assay buffer. For concentration response curves, compounds were serially diluted 3-fold (from 2 mM top concentration to 0.1013 mM) to generate a 10-point curve and 125 nL was transferred to the assay plate using an Echo sonic dispenser, with a final concentration range of 26.6 μM to 1.35 μM in 10 μL of reaction. Five microliters of a 10 nM PARP1 enzyme and 100 nM core histone (2x) mixture were added to the assay plate and pre-incubated for 30 min at room temperature (RT). The reaction was initiated by adding 5 μL of a 10 μM β-NAD plus 0.02 mg / mL activated DNA (2X) mixture. The reaction was incubated for 60 min at RT. The final concentrations of PARP1 enzyme and substrate were 5 nM and 5 μM, respectively. Positive control (high signal) wells and negative control (low signal) wells were replaced with 125 nL DMSO instead of compound. After incubation, the reaction was terminated by adding 10 μL of 10 μM 3ABA PARP1 inhibitor, incubated for 5 min at RT, and placed in a -80 °C freezer for shipment to the Valo Health site in Branford, CT, where mass spectrometry will be used to directly measure nicotinamide (NAM) levels.

[0314] The percentage inhibition of enzyme activity was calculated according to the following equation:

[0315]

[0316] The S in the equation 样品 , S 高 and S 低 The values ​​refer to the NAM concentrations detected in the assay wells, high control wells, and low control wells. To determine the inhibitor IC 50 Values ​​were used and the % inhibition of CRC (concentration response curves) were fit to a standard one-site four-parameter logistic equation.

[0317] Representative results for exemplary compounds of the present technology

[0318] Table 1 provides representative initial results for exemplary compounds of the present technology.

[0319] Table 1.

[0320]

[0321]

[0322]

[0323]

[0324] PAR-based immunofluorescence assay protocol Materials:

[0325]

[0326]

[0327] equipment:

[0328] Device Name supplier Multidrop Combi ThermoFisher Echo 555 Labcyte / Beckman Coulter Life Sciences Cellometer Auto T4 Brightfield Cell Counter Nexcelom Bioscience Bluewasher Blue Cat Bio PlateLoc Microplate Heat Sealer Agilent CX7 Thermofisher

[0329] Cell culture details: HCT116 is a human colon epithelial cell line that grows adherently in tissue culture flasks. Grow cells in T175 size flasks. Split twice a week at 70-80% confluence, split ratio 1:5-1:10, following standard adherent cell subculture protocols. Keep media refrigerated until the day of cell-based activities and need to be warmed to at least RT before use. Cell culture medium: McCoy 5A with 10% FBS, 2mM L-glutamine, 20mM HEPES. Plating medium: McCoy 5A with 10% FBS, 2mM L-glutamine, 20mM HEPES, 1% antibiotic antimycotic solution.

[0330] program:

[0331] Day 1: Cell culture and cell plating: The cell culture and cell plating method includes the following steps: warming the culture medium before cell culture; removing the cell flask from the incubator and trypsinizing the cells in the T175 flask using 0.25% trypsin; returning the flask to the incubator for about 5 minutes to allow the cells to fall from the bottom of the flask; adding 10 μL of culture medium to rinse the bottom of the flask, and adding the cells and culture medium to a 50 mL conical tube; spinning the cells at 1000 rpm for 5 minutes, aspirating the culture medium, and resuspending the pellet with 10 mL of fresh culture medium; using Ce llometer cell counter to calculate viable cell count; using the viable cell count and the number of plates required, calculate the amount of medium to resuspend the cell pellet, which needs to be added to the fresh plating medium to be dispensed into the 384W plate at a density of 5 μL / well at 6000 cells / well; plate the cells in the 384W plate at 5 μL of medium per well at 6000 cells / well for the entire plate; gently shake the plate (100 rpm) for 20 min after plating; and incubate the plate at 37°C, 5% CO before processing. 2 Incubate overnight.

[0332] Day 2: Compound and DNA damage treatment, fixation and primary staining:The method includes the following steps: 384W cell seeding plates are processed using a Labcyte Echo 555 to achieve a top dose of 10 μM for each compound. 40 nL of the 10 mM top dose compound plate is stamped into 40 μL of cells in the seeding plate. The Labcyte compound source plate is based on DMSO and compounds are diluted in ten three-fold doses starting at 10 mM. The transfer volume using the Echo is 40 nL; the cells are incubated at 37°C, 5% CO 2 Incubate for 5 hours at 37°C and 5% CO; add the DNA damaging agent MMS to columns 1-23 of each cell-seeded 384W plate by delivering 40 nl of 19.2% MMS in DMSO using Echo in DMSO delivery mode, where the MMS stock solution concentration is 99% and must be diluted 1:5 in DMSO before delivery to the cell plate; incubate the cell plate at 37°C, 5% CO 2 Incubate for 30 minutes at 4 °C; evacuate the medium using BlueWasher and add 75 μl cold methanol, with the Bluewasher MagBeadSpeed ​​setting for all evacuations: rotate the plate clockwise at 800 RPM (35g) for 5 seconds; keep the dispensing lines and methanol cold throughout the fixation process, and use the intermittent setting to divide the dispensing into multiple injections to minimize cell disruption; incubate the fixed plate on ice for 20 minutes; wash the entire plate 1x with an equal volume of cold DPBS and evacuate with Bluewasher at MagBeadSpeed; add 20 μl of DPBS 0.1% Triton X-100 to the entire plate and incubate at room temperature for 15 minutes, and evacuate with Bluewasher at MagBeadSpeed; add 20 μl of Roche blocking reagent to the entire plate and incubate at room temperature for 60 minutes, and evacuate with Bluewasher at MagBeadSpeed; and add 20 μl of Roche blocking reagent PAR antibody (1:4,000), seal the plate and incubate overnight at 4°C.

[0333] Day 3: Secondary Antibody Staining and Imaging:The method includes the following steps: evacuate using Bluewasher and wash the plate (3X) with 25-30μl DPBS 0.05% Tween 20; add 20μl anti-mouse AF488 secondary antibody (1:1,600) plus Hoechst (1:10,000) and incubate at room temperature for 60 min; evacuate using MagBeadSpeed ​​and wash 3X to 4X with 25-30μl DPBS 0.05% Tween 20; add 30μl DPBS and seal the plate; and image using the protocol "PAR_HCT116_MeOH_10X_2ChR" using CX7-round (nuclei) medium average intensity channel 1 (360) and channel 2 (488). Key acquisition settings: Expose the 488 channel to 70% to 80% and 1 to 2 fields.

[0334] Data Analysis: Raw data files are exported from the CX7 software and paired with barcoded compound plates to track compound IDs, dose responses, and Echo transmission records. This allows for the import and QC analysis of dose response curves, as well as IC50 value determination. Images of each plate and well on both imaging channels can also be exported. The percentage inhibition of PAR-based activity is calculated according to the following equation:

[0335]

[0336] The S in the equation 样品 , S 高 and S 低 The values ​​refer to the PARylation levels detected in the assay wells, high control wells, and low control wells, respectively. To determine the inhibitor IC 50 Values ​​were used and the % inhibition of CRC (concentration response curves) were fit to a standard one-site four-parameter logistic equation.

[0337] Determination principle: The cellular levels of PARylation of PARP1 / 2 substrate proteins (PARP1 and histones) were measured by using a DNA damage immunofluorescence assay with anti-PAR antibodies based on https: / / f1000research.com / articles / 5-736 / v2. PARylation levels were reduced when PARP inhibitors were co-treated with the DNA damaging agent MMS.

[0338] Cell-based proliferation assay protocols: DLD1 parental vs. BRCA2 null 5-day CTG, CTF, CyQuant or one-pot (One Pot) Live-Dead HCS Assay

[0339] Reagents and consumables:

[0340]

[0341]

[0342] equipment:

[0343] Device Name supplier Multidrop Combi ThermoFisher Echo 555 Labcyte / Beckman Coulter Life Sciences Cellometer Auto T4 Brightfield Cell Counter Nexcelom Bioscience Bluewasher Blue Cat Bio PlateLoc Microplate Heat Sealer Agilent CX7 ThermoFisher PHERAstar FS BMG Labtech EnVision PerkinElmer

[0344] program

[0345] Day 1: Cell culture and cell plating: The method includes the following steps: count cells (using a Nexcelom cell counter and record the cell count); spin cells at 1000 rpm for 5 min and resuspend the pellet with fresh medium; and plate cells in a 1536-well plate at a specific cell / well density based on density optimization testing (or 100 cells / well for DLD1 parental (historically); 125 cells / well for DLD1 BRCA2+ / - and - / - null; μL of cells / well in columns 1 to 47 for 1536w plates); gently shake the plate (100 rpm) at room temperature for 30 min before transferring to an incubator (37°C, 5% CO 2 ) and the plates (37°C, 5% CO 2 ) and incubate for 24h.

[0346] Day 2: Compound Treatment: For 1536w, the highest concentration of source plate compound was compound stamped at 25nl 10 mM, resulting in a highest dose concentration of 50 μM for cell based proliferation assays. The positive control SAHA was stamped at 10 μM final concentration.

[0347] Day 3-7: Plate incubation: After adding the compounds, the plates were incubated at 37°C, 5% CO 2 Incubate at 4 °C for a total of 120 h / 5 days.

[0348] Day 3-7: Readout reagent additions and plate readings: After incubation, remove the plate from the incubator and add the designated readout reagent to the cell plate. See below for details based on the specific readout.

[0349] CTG reading: Remove CTG 2.0 reagent from refrigerator and return to room temperature before use; add 4 μL / well CTG to all wells of each cell plate; for 1536w plates, incubate at 37°C, 5% CO 2 Incubate the plate for 30 minutes at 4 °C; and read the plate on a designated plate reader using a luminescence-specific protocol; BMG or Envision have a CTG-specific protocol.

[0350] ●CyQUANT Reading: Remove the Cyquant Direct Cell Proliferation Assay Kit from the refrigerator and return to room temperature before use. The reagent may need to be heated in a dry bath to thaw; calculate the total volume required based on the amount dispensed to ensure that enough assay reagent is required. Prepare the assay reagent in combination with the following assay recipe: PBS:

[0351] 11.7mL; Direct Nucleic Acid Stain: 48 μL; and Direct Background Suppressor:

[0352] 240 μL; 2 μl / well Cyquant reagent mixture was added to all wells; the plate was incubated at 37°C, 5% CO 2 Incubate for 60 minutes at 4 °C; read the plate on a designated plate reader BMG or EnVisions using the CyQuant specific protocol with the specific 1536w protocol (CyQUANTDirect—508 / 527 nm; CyQUANTDirect Red—622 / 645 nm).

[0353] CTF reading: Before use, remove the CTF reagent from the refrigerator and return to room temperature. Prepare 1X reagent and vortex to dissolve (reagent is stable for 24 hours at RT or 7 days at 4°C). Add 4 μl / well CTF to all wells and incubate at 37°C, 5% CO 2 Incubate the plate for 180 minutes (3 h) at 4 °C; and read the plate on a designated plate reader using a fluorescence specific protocol (BMG or Envisions have a CTF specific protocol (Ex 380-400; Em 505)).

[0354] ● One-pot Live-Dead HCS Reading: After incubation, remove the plate from the incubator; for a 1536-well plate, prepare 4 mL

[0355] 1X PBS and add 8 drops of propidium iodide reagent + 4 μL Hoechst 333242; mix, and then dispense at 1 μL / well at medium speed using a Multidrop Combi; For 384-well plates: prepare 6 mL of 1X PBS and add 12 drops of propidium iodide reagent + 6 μL Hoechst 333242, mix, and then dispense at 8 μL / well at medium speed using a Multidrop Combi; incubate at 37°C, 5% CO 2 Incubate at 4 °C for 30 min; seal the plate with an aluminum seal;

[0356] Spin at 1000 RPM for 2 minutes in a swing bucket centrifuge; load the plate onto the CX7; and acquire data.

[0357] The analyzed data were then exported as ignition files and subsequently processed in ABASE.

[0358] Data analysis of CTG, CyQUANT and CTF reads: Raw data files are exported from BMG, Envision and CX7 readers, paired with barcoded compound plates to track compound IDs, dose responses and Echo transmission records. This allows for the import and QC analysis of dose response curves, as well as IC50 value determination. Data and images for each plate and well on both imaging channels can also be exported. The percentage inhibition of growth activity is calculated according to the following equation:

[0359]

[0360] The S in the equation 样品 , S 高 and S 低 The values ​​refer to the assay signal detected in the wells, high control wells, and low control wells, respectively. To determine the inhibitor IC 50 Values ​​were used and the % inhibition of CRC (concentration response curves) were fit to a standard one-site four-parameter logistic equation.

[0361] Determination principle: Cell viability was measured using assays that detect the following live cell readouts: intracellular ATP (CTG), DNA (CyQUANT), and peptidase activity (CTF) in the presence of the cell permeable substrate Gly-Phe-AFC using luminescent or fluorometric assays following cell proliferation in the presence of PARP inhibitor compounds.

[0362] MDCK-MDR1 Permeability Assay Results for Exemplary Compounds of the Present Technology

[0363] The bidirectional assay (which contains a Mandin Darby canine kidney (MDCK) cell line transfected with the human MDR1 gene (P-glycoprotein; P-gp), which is designed to overexpress the MDR1 efflux transporter, a major efflux transporter at the blood-brain barrier (BBB)) uses an established method to measure the rate of flux of a compound across a polarized cell monolayer. As is well known in the art, data generated from the MDCK-MDR1 permeability assay can be used to predict the in vivo absorption of a drug. The bidirectional MDCK-MDR1 permeability assay can identify and quantify the level of active efflux. Compounds that pass through the cell monolayer in both the apical to basolateral direction (A→B) and the basolateral to apical direction (B→A) are screened, providing a ratio of B→A / A→B (efflux ratio; "ER"). Unlike the Caco-2 assay, the MDCK-MDR1 assay is not subject to potential efflux interference from breast cancer resistance protein (BCRP). When a compound has an ER greater than 2, it indicates that the compound may undergo active efflux. The data can be further used to predict blood-brain barrier (BBB) ​​permeability via calculations from the log(permeability-surface area product) ("logPS") data, which is well known to predict in vivo log([brain concentration] / [plasma concentration]).

[0364] Table 2 below provides initial bidirectional MDCK-MDR1 permeability results for exemplary compounds of the present technology.

[0365] Table 2.

[0366]

[0367]

[0368]

[0369] In Vivo CNS Penetration of Exemplary Compounds of the Technology

[0370] According to the internal proprietary prediction model, the compound of the present technology is predicted to show BBB passage. These prediction models were further verified by in vivo experiments. In particular, the free drug concentration in the brain and plasma after continuous intravenous infusion in Sprague Dawley rats for 24h (corrected protein binding) was compared to measure the unbound brain to plasma ratio (Kpuu) in vivo at steady state, where Kpuu>0.3 indicates good brain exposure of the compound.

[0371] Table 3 below provides Kpuu results for exemplary compounds of the present technology. As illustrated in Table 3, compounds of the present technology exhibited in vivo Kpuu values ​​that were generally acceptable for predicting clinical brain penetrance.

[0372] Table 3.

[0373] Compound Kpuu 0114B 0.921 0174B 0.591

[0374] Although certain embodiments have been illustrated and described, those skilled in the art, after reading the foregoing description, may make changes, equivalent substitutions and other types of changes to the compounds or salts thereof, pharmaceutical compositions, derivatives, prodrugs, metabolites, tautomers or racemic mixtures of the present technology as described herein. Each of the above aspects and embodiments may also have been included or incorporated therein, such as variations or aspects disclosed with respect to any or all other aspects and embodiments.

[0375] The present technology is also not limited to the specific aspects described herein, which are intended to be a single illustration of the individual aspects of the present technology. Without departing from the spirit and scope of the present technology, many modifications and changes can be made to the present technology, as will be apparent to those skilled in the art. In addition to the methods listed herein, it will be apparent to those skilled in the art from the above description that the functionally equivalent methods within the scope of the present technology. Such modifications and changes are intended to fall within the scope of the appended claims. It should be understood that the present technology is not limited to specific methods, reagents, compounds, compositions, labeled compounds or biological systems, which of course can vary. It should also be understood that the terms used herein are only for the purpose of describing specific aspects, and are not intended to be restrictive. Therefore, it is hoped that this specification is only regarded as exemplary, wherein the breadth, scope and spirit of the present technology are indicated only by the appended claims, the definitions therein and any equivalents thereof.

[0376] The embodiments described illustratively herein can be appropriately practiced in the absence of any one or more elements, one or more limitations not specifically disclosed herein. Thus, for example, the terms "include", "comprise", "contain", etc. should be read broadly without limitation. In addition, the terms and expressions adopted herein have been used as terms of the specification and are not limited, and the use of such terms and expressions is not intended to exclude any equivalents or parts of the features shown and described, but it should be recognized that various modifications can be made within the scope of the claimed technology. In addition, the phrase "essentially consisting of..." will be understood to include those elements specifically described and those other elements that do not substantially affect the basic and novel features of the claimed technology. The phrase "consisting of..." excludes any unspecified elements.

[0377] In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group. Each of the narrower classes and subgeneric groupings falling within the generic disclosure also form part of the present invention. This includes the generic description of the invention with the proviso or negative limitation that any subject matter is removed from the genus, regardless of whether the deleted material is specifically described herein.

[0378] As will be understood by those skilled in the art, for any and all purposes, particularly in providing a written description, all ranges disclosed herein also encompass any and all possible subranges and combinations of subranges thereof. Any listed range can be easily considered to fully describe and enable the same range to be decomposed into at least equal halves, thirds, quarters, fifths, tenths, etc. As a non-limiting example, each range discussed herein can be easily decomposed into a lower third, a middle third, and an upper third, etc. As will be equally understood by those skilled in the art, all languages ​​such as "at most," "at least," "greater than," "less than," etc. include the numbers listed and refer to the ranges that can subsequently be decomposed into subranges as discussed above. Finally, as will be understood by those skilled in the art, a range includes each individual member.

[0379] All publications, patent applications, issued patents, and other documents (e.g., journals, articles, and / or textbooks) cited in this specification are incorporated herein by reference as if each individual publication, patent application, issued patent, or other document was specifically and individually indicated to be incorporated by reference in its entirety. Definitions contained in text incorporated by reference are excluded when definitions in the text are inconsistent with definitions in this disclosure.

[0380] The present technology may include, but is not limited to, the features and combinations of features listed in the following lettered paragraphs, with the understanding that the following paragraphs shall not be interpreted as limiting the scope of the appended claims or mandating that all such features must be included in these claims:

[0381] A. A compound of formula I

[0382]

[0383] or a pharmaceutically acceptable salt and / or solvate thereof, wherein

[0384] X 1 is H, F or Cl;

[0385] X 2 Is NH or NR 3 ;

[0386] R1 and R 2 One of is H, halo, alkyl, alkenyl, alkynyl, cycloalkyl or heterocyclyl, and R 1 and R 2 The remaining one of is aryl, heteroaryl or non-aromatic heterocyclic group; and

[0387] R 3 It is an alkyl group, a cycloalkyl group, an alkylene group and a non-aromatic heterocyclic group.

[0388] B. A compound according to paragraph A, wherein the compound is a compound of formula IA

[0389]

[0390] or a pharmaceutically acceptable salt and / or solvate thereof.

[0391] C. A compound according to paragraph A, wherein the compound is a compound of formula (IB)

[0392]

[0393] or a pharmaceutically acceptable salt and / or solvate thereof.

[0394] D. A compound according to any one of paragraphs A to C, wherein

[0395] R 1 is aryl, heteroaryl or non-aromatic heterocyclic; and

[0396] R 2 is H, halo, alkyl, alkenyl, alkynyl, cycloalkyl or non-aromatic heterocyclyl. E. A compound according to any one of paragraphs A to D, wherein

[0397] R 1 is aryl, heteroaryl or non-aromatic heterocyclic; and

[0398] R 2 is H, alkyl, cycloalkyl or non-aromatic heterocyclic.

[0399] F. A compound according to any one of paragraphs AE, wherein X 1 It's F.

[0400] G. A compound according to any one of paragraphs AF, wherein X 2 It's NH.

[0401] H. A compound according to any one of paragraphs AG, wherein the compound is

[0402]

[0403]

[0404]

[0405]

[0406]

[0407]

[0408]

[0409]

[0410]

[0411]

[0412]

[0413]

[0414]

[0415] or a pharmaceutically acceptable salt and / or solvate of any one thereof.

[0416] I. A composition comprising: a compound according to any one of paragraphs A to H; and a pharmaceutically acceptable carrier.

[0417] J. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and an effective amount of a compound according to any one of paragraphs A to H, wherein the effective amount of the compound is effective for treating cancer.

[0418] K. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a compound according to any one of paragraphs A to H, wherein the compound is present in an amount effective to treat cancer when combined with a second cancer therapy.

[0419] L. A method of treating a subject suffering from a B-cell malignancy, the method comprising administering to the subject an effective amount of a compound according to any one of paragraphs A to H and an effective amount of a second cancer therapy.

[0420] M. A medicament for treating cancer in a subject, the medicament comprising a compound according to any one of paragraphs A to H.

[0421] N. The medicament according to paragraph M, wherein the medicament further comprises a pharmaceutically acceptable carrier.

[0422] O. The medicament according to paragraph M or paragraph N, wherein the medicament comprises an effective amount of the compound for treating the cancer when combined with a second cancer therapy.

[0423] Other embodiments are set forth in the following claims, along with the full scope of equivalents to which such claims are entitled.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt and / or solvate thereof, wherein X 1 is H, F or Cl; X 2 Is NH or NR 3 ; R 1 and R 2 One of is H, halo, alkyl, alkenyl, alkynyl, cycloalkyl or heterocyclyl, and R 1 and R 2 The remaining one of is aryl, heteroaryl or non-aromatic heterocyclic group; and R 3 It is an alkyl group, a cycloalkyl group, an alkylene group and a non-aromatic heterocyclic group.

2. The compound according to claim 1, wherein R 1 is aryl, heteroaryl or non-aromatic heterocyclic; and R 2 is H, halo, alkyl, alkenyl, alkynyl, cycloalkyl or non-aromatic heterocyclyl.

3. The compound according to claim 1, wherein R 1 is aryl, heteroaryl or non-aromatic heterocyclic; and R 2 is H, alkyl, cycloalkyl or non-aromatic heterocyclic.

4. The compound according to claim 1, wherein X 1 It's F.

5. The compound according to claim 1, wherein X 2 It's NH.

6. The compound according to claim 1, wherein the compound is a compound of formula IA or a pharmaceutically acceptable salt and / or solvate thereof.

7. The compound according to claim 6, wherein R 1 is aryl, heteroaryl or non-aromatic heterocyclic; and R 2 is H, halo, alkyl, alkenyl, alkynyl, cycloalkyl or non-aromatic heterocyclyl.

8. The compound according to claim 7, wherein R 1 is aryl, heteroaryl or non-aromatic heterocyclic; and R 2 is H, alkyl, cycloalkyl or non-aromatic heterocyclic.

9. The compound according to claim 8, wherein X 1 It's F.

10. The compound according to claim 9, wherein X 2 It's NH.

11. The compound according to claim 1, wherein the compound is a compound of formula IB or a pharmaceutically acceptable salt and / or solvate thereof.

12. The compound according to claim 11, wherein R 1 is aryl, heteroaryl or non-aromatic heterocyclic; and R 2 is H, halo, alkyl, alkenyl, alkynyl, cycloalkyl or non-aromatic heterocyclyl.

13. The compound according to claim 12, wherein R 1 is aryl, heteroaryl or non-aromatic heterocyclic; and R 2 is H, alkyl, cycloalkyl or non-aromatic heterocyclic.

14. The compound according to claim 13, wherein X 1 It's F.

15. The compound according to claim 14, wherein X 2 It's NH.

16. A composition comprising: a compound according to any one of claims 1 to 15; and a pharmaceutically acceptable carrier.

17. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and an effective amount of a compound according to any one of claims 1 to 15, wherein the effective amount of the compound is effective for treating cancer.

18. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a compound according to any one of claims 1 to 15, wherein the compound is present in an amount effective to treat cancer when combined with a second cancer therapy.

19. A method of treating a subject suffering from a B-cell malignancy, the method comprising administering to the subject an effective amount of a compound according to any one of claims 1 to 15 and an effective amount of a second cancer therapy.

20. A medicament for treating cancer in a subject, the medicament comprising a compound according to any one of claims 1 to 15. The medicament according to claim 20 , wherein the medicament further comprises a pharmaceutically acceptable carrier.

22. The medicament of claim 20, wherein the medicament comprises an effective amount of the compound for treating the cancer when combined with a second cancer therapy.

23. The medicament of claim 21, wherein the medicament comprises an effective amount of the compound for treating the cancer when combined with a second cancer therapy.

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