Inhibitors of MYC and their use

Abstract

A substituted heterocyclic compound and a proteolysis-targeting chimera molecule (PROTAC) are disclosed. The substituted heterocycles disclosed herein are shown to be useful for inhibiting c-MYC. The disclosed PROTACs have been shown to induce the degradation of c-MYC protein. The substituted heterocyclic compounds and proteolysis-targeting chimera molecules (PROTACs) disclosed herein can be utilized as therapeutic agents for treating cancer and cell proliferative disorders.

Description

Technical Field

[0001] (Cross - Reference to Related Patent Applications) This application claims the benefit of priority of U.S. Patent Application No. 63 / 369,892, filed on July 29, 2022, the content of which is incorporated herein by reference in its entirety.

[0002] Description of Research or Development Sponsored by the Federal Government This invention was made with government support under grant numbers CA180995 and CA257258 awarded by the National Institutes of Health. The government has certain rights in this invention.

Background Art

[0003] The field of the present invention relates to substituted heterocycles as c - MYC targeting agents and proteolysis - targeting chimeric molecules (PROTACs) that induce the degradation of c - MYC protein. In particular, the field of the present invention relates to substituted pyrazoles, imidazoles or triazoles as c - MYC targeting agents, and PROTACs that target c - MYC for degradation, which can be used for the treatment of cell proliferation diseases and disorders such as cancer.

[0004] The c-MYC oncogene is deregulated and plays a causal role in most human cancers, and c-MYC inhibition significantly affects tumor growth or survival in multiple models. MYC is the most common oncogene involved in human cancers and is overexpressed in up to half of all cancers. Therefore, the development of c-MYC inhibitors is one of the most attractive potential anti-cancer strategies. Unfortunately, c-MYC is currently considered "undruggable" because it is difficult to target transcription factors with small molecules. Compounds that selectively target c-MYC-driven cell proliferation and interfere with c-MYC binding to DNA can be found, for example, in U.S. Patent Application Publication No. 2017 / 0253581, published on September 7, 2017, U.S. Patent Application Publication No. 2019 / 0062281, published on February 28, 2019, U.S. Patent Application Publication No. 2020 / 0390894, published on December 17, 2020, and U.S. Patent Application Publication No. 2020 / 0392116, published on December 17, 2020, the contents of which are hereby incorporated by reference in their entirety.

[0005] Proteolysis-targeting chimeric molecules (PROTACs) are an emerging technology that can be used to target previously "undruggable" targets such as transcription factors and non-enzyme proteins. (See, for example, An et al., "Small-molecule PROTACs: An emerging and promising approach for the development of targeted therapy drugs," EBioMedicine. 2018 Oct;36:553-562; and Gu et al., "PROTACs: An Emerging Targeting Technique for Protein Degradation in Drug Discovery," Bioessays. 2018 Apr;40(4):e1700247, the contents of which are hereby incorporated by reference in their entirety). PROTACs are chimeric molecules that can be characterized as "heterobifunctional" in that they contain a ligand for recruiting an E3 ubiquitin ligase, a linker, and another ligand that binds to the protein to be targeted for degradation. When designed in this way, PROTACs "hijack" the E3 ubiquitin ligase to the protein targeted for proteolysis via ubiquitination, even when the target protein is not a physiological substrate for degradation via the ubiquitin-proteasome system.

[0006] Here, the inventors disclose an approach that selectively targets c-MYC-driven cell proliferation and interferes with the binding of c-MYC to DNA. PROTACs that induce the degradation of the c-MYC protein are also disclosed. SUMMARY OF THE INVENTION

[0007] Substituted heterocycles that can be used as c-MYC targeting agents are disclosed. The substituted heterocycles can include substituted pyrazoles, substituted imidazoles, and substituted triazoles. The disclosed heterocycles can be used in pharmaceutical compositions and methods for treating cell proliferative disorders such as cancer.

[0008] The disclosed substituted complex rings may include substituted pyrazoles, imidazoles, and triazoles having the formula I, [Chemical formula] wherein, W is CR 8 or N, Y is CH or N, Q is CR 2 or N, Z is C(Alk 2 ) q (X) p (Alk 1 ) n R 1 or N, R 1 is hydrogen, halo, alkyl, aryl, benzyl, heteroaryl, cycloalkyl, alkoxy or cycloheteroalkyl, R 1 is optionally substituted at one or more positions with one or more of alkyl, alkoxy, cycloalkyl, haloalkyl, haloalkoxy, aryl, hydroxyl, halo, cyano, amide, carboxyl, -(CH2)-NH-R 9 , -(O(CH2)2) m -R 15 , or -OR 11 , Alk 1 is straight-chain or branched-chain alkenylenyl, or cycloalkenylenyl, n is 0, 1 or 2, p is 0 or 1, Alk 2 is straight-chain or branched-chain alkenylenyl, q is 0 or 1, r is 0 or 1, m is an integer selected from 1 to 20, X is O or NR 13 , R 2 is hydrogen or halo, R 3is alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyl, -O-C(O)-alkyl, or halo, R 4 is hydrogen, halo, amino, alkyl or haloalkyl, or R 4 is aryl or benzyl optionally substituted at one or more ring positions with one or more alkyl, alkoxy, haloalkyl, haloalkoxy, aryl, hydroxyl, halo, cyano, amide, carboxyl, aryloxy, and alkylaryloxy, or R 4 is alkyl optionally substituted with halosubstituted aryloxy, or R 4 is R 1 together with R forms a cycloheteroalkyl fused to ring A, and the cycloheteroalkyl fused to ring A is optionally substituted with aryl or alkylaryl substituted with halogen, R 5 is hydrogen, halo, alkyl, aryl, alkylaryl, heteroaryl, cycloalkyl or cycloheteroalkyl, and optionally, R 5 is substituted at one or more positions with one or more alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyl, halo, cyano, carboxamide, carboxyl, aryloxy and heteroaryloxy, R 6 is hydrogen, halo, alkyl, aryl, alkylaryl, heteroaryl, cycloalkyl or cycloheteroalkyl, and optionally, R 6 is substituted at one or more positions with one or more alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyl, halo, cyano, carboxamide, carboxyl, aryloxy and heteroaryloxy, R 7 is alkyl, R 8 is hydrogen, cyano, amino, alkyl, alkoxy, haloalkyl, haloalkoxy, aryl, benzyl, hydroxyl, halo, amide, and carboxyl, R 9 is one or more -P(O)(OR10 )2 is alkyl optionally substituted with R 10 is hydrogen or alkyl, R 11 is alkyl optionally substituted with one or more -P(O)(OR 12 )2, R 12 is hydrogen or alkyl, R 13 is hydrogen, alkyl or -C(O)R 14 , R 14 is aryl optionally substituted at one or more ring positions with one or more alkyl, alkoxy, haloalkyl, haloalkoxy, aryl, hydroxyl, halo, cyano, amide and carboxyl, R 15 is OR 16 , -OS(O)2-R 16 , or NR 17 R 18 , R 16 is alkyl or aryl, and R 16 is optionally substituted at one or more positions with one or more alkyl, alkoxy, haloalkyl, haloalkoxy, aryl, hydroxyl, halo, cyano, amide and carboxyl, R 17 and R 18 are independently hydrogen or alkyl.

[0009] The disclosed compounds may exhibit one or more biological activities. The disclosed compounds may inhibit the binding of the Myc / Max complex to DNA (e.g., in a DNA gel shift assay). The disclosed compounds may not cause significant DNA damage (e.g., in an rH2AX staining assay, at concentrations of about 0.001 μM, 0.005 μM, 0.01 μM, 0.1 μM, 1.0 μM, 10 μM, 100 μM or greater). The disclosed compounds may inhibit the growth of cells expressing c-MYC (preferably at concentrations of less than about 100 μM, 50 μM, 10 μM, 1 μM, 0.1 μM, 0.05 μM, 0.01 μM, 0.005 μM, 0.001 μM or less). The disclosed compounds may not inhibit the growth of cells not expressing c-MYC (preferably at concentrations greater than about 0.001 μM, 0.005 μM, 0.01 μM, 0.5 μM, 0.1 μM, 1.0 μM, 10 μM, and 100 μM).

[0010] Also disclosed are proteolysis targeting chimeric molecules (PROTACs) that induce the degradation of the c-MYC protein. The disclosed PROTACs include a portion that binds to c-MYC covalently attached to a portion that binds to a ubiquitin ligase. The disclosed PROTACs may typically include a first targeting moiety that binds to c-MYC(M c-MYC ) and may be derived from a substituted heterocycle that binds to c-MYC, such as a substituted pyrazole. The first targeting moiety may be covalently attached, via a bond or a linker (L), to a second targeting moiety that binds to a ubiquitin ligase such as an E3 ubiquitin ligase (M E3 ). Accordingly, the disclosed PROTACS may be described as having Formula II or Formula M c-MYC -L-M E3 or M E3 -L-M c-MYC .

[0011] The disclosed PROTACs target the E3 ubiquitin ligase moiety to c-MYC, which is then ubiquitinated and targeted for degradation. The disclosed PROTACs may be used in the treatment of diseases and disorders associated with c-MYC, such as cell proliferation diseases and disorders including cancer.

[0012] The disclosed PROTAC typically includes a first targeting moiety that binds to c-MYC(M c-MYC ), which is derived from a substituted heterocyclic ring that binds to c-MYC. Suitable substituted heterocyclic rings that bind to c-MYC may include, but are not limited to, substituted pyrazoles.

[0013] The c-MYC targeting moiety of the disclosed PROTAC(M c-MYC ) is typically linked via a linker or linker (L) to a second targeting moiety that binds to an E3 ubiquitin ligase(M E3 ).

[0014] Suitable linkers for the disclosed PROTAC may include, but are not limited to, linkers that include a polyethylene glycol moiety.

[0015] The E3 ubiquitin ligase targeting moiety of the disclosed PROTAC(M E3 ) typically binds and / or targets the PROTAC to an E3 ubiquitin ligase. Suitable E3 ubiquitin ligases may include, but are not limited to, the Von Hippel-Lindau (VHL) E3 ubiquitin ligase, the cereblon (CRBN) E3 ubiquitin ligase, the inhibitor of apoptosis protein (IAP) E3 ubiquitin ligase, and the murine double minute 2 homolog (MDM2) E3 ubiquitin ligase.

[0016] The E3 ubiquitin ligase targeting moiety of the disclosed PROTAC(M E3 ) typically is derived from a compound that binds to the E3 ubiquitin ligase, for example, as a ligand of the E3 ubiquitin ligase. Suitable ligands may include, but are not limited to, thalidomide, pomalidomide, lenalidomide, VHL ligand 1 (VHL-1), VHL ligand 2 (VHL-2), VH032, VL-269, LCL161, hydroxyproline-based ligands, and ligands derived from HIF-1α-derived (R)-hydroxyproline including radicalized forms.

[0017] The disclosed PROTACs may exhibit one or more biological activities. The disclosed PROTACs may inhibit the growth of cells expressing c-MYC (preferably at a concentration of less than about 100 μM, 50 μM, 10 μM, 1 μM, 0.1 μM, 0.05 μM, 0.01 μM, 0.005 μM, 0.001 μM or less). The disclosed PROTACs cannot inhibit the growth of cells that do not express c-MYC (preferably at concentrations greater than about 0.001 μM, 0.005 μM, 0.01 μM, 0.5 μM, 0.1 μM, 1.0 μM, 10 μM, and 100 μM).

[0018] In some embodiments, the PROTAC has the formula: M c-MYC -L-M E3 (wherein M c-MYC is a moiety that binds to c-MYC, and L is a bond or linker that covalently attaches M MYC and M E3 and M E3 is a moiety that binds to an E3 ubiquitin ligase). In some embodiments, the PROTAC has the formula II: [Chemical formula] and has wherein R 1 is hydrogen, cyano, amino, alkyl, alkoxy, haloalkyl, haloalkoxy, aryl, benzyl, hydroxyl, halo, amide, and carboxyl, R 2 is alkyl, R 3 is alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyl, -O-C(O)-alkyl, or halo, R 4 is hydrogen, halo, alkyl, aryl, alkylaryl, heteroaryl, cycloalkyl or cycloheteroalkyl, and optionally, R 4is substituted at one or more positions with one or more of alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyl, halo, cyano, carboxamide, carboxy, aryloxy and heteroaryloxy, X is O or NR 5 wherein, R 5 is hydrogen, alkyl or -C(O)R 6 wherein, R 6 is aryl optionally substituted at one or more ring positions with one or more of alkyl, alkoxy, haloalkyl, haloalkoxy, aryl, hydroxyl, halo, cyano, amide and carboxyl, Y is alkenylenyl, arylenylenyl, benzylenylenyl, heteroarylenylenyl, cycloalkenylenyl or cycloheteroalkenylenyl, L is a bond, or -(O(CH2)2) a -, -(O(CH2)2) a -NH-C(O)-Alk 3 -, -(O(CH2)2) a -C(O)-NH-((CH2)2-O) b -Alk 3 -, -(O(CH2)2) a -C(O)-NH-Alk 3 -C(O)-, -(O(CH2)2) a -C(O)-NH-((CH2)2-O) b -, -(O(CH2)2) a -C(O)-NH-((CH2)2-O) b -Alk 3 -C(O)-, -(O(CH2)2) a -C(O)-NH-Alk 3 -NH-C(O)-CH2-, and -(O(CH2)2) a -C(O)-NH-Alk 3 is a linker selected from the group consisting of a is an integer selected from 1 to 20, b is an integer selected from 1 to 20, Alk 3is a linear or branched alkenylenyl, M E3 is [Chemical formula] selected from the group consisting of R 7 is alkyl optionally substituted with one or more amino or -S(O)2R 8 wherein R 8 is alkyl or aryl, and R 8 is optionally substituted at one or more positions with one or more alkyl, alkoxy, haloalkyl, haloalkoxy, aryl, hydroxyl, halo, cyano, amide, and carboxyl, R 9 R 10 and R 11 are independently hydrogen or alkyl.

[0019] Also disclosed are pharmaceutical compositions comprising the disclosed compounds and / or the disclosed PROTACs and a suitable pharmaceutical carrier, excipient, or diluent. The disclosed pharmaceutical compositions may comprise an effective amount of a compound and / or a PROTAC for inhibiting the growth of cancer cells when administered to a subject in need thereof.

[0020] Also disclosed is a method of treating cell proliferation diseases and disorders such as cancer. The method may comprise administering the disclosed compound, and / or the disclosed PROTAC, or a pharmaceutical composition comprising the disclosed compound to a subject in need thereof, such as a subject having cancer. The disclosed compound and / or the disclosed compound and / or a PROTAC or a pharmaceutical composition comprising the disclosed compound and / or a PROTAC may be administered optionally in combination with additional therapeutic agents for treating cell proliferation diseases and disorders. Cell proliferation diseases and disorders treatable by the disclosed method may include, but are not limited to, cancers selected from the group consisting of multiple myeloma, leukemia, non-small cell lung cancer, colon cancer, central nervous system cancer, melanoma, ovarian cancer, renal cancer, prostate cancer, and breast cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0021]

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[0022]

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[0023]

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[0024]

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[0025]

Figure 12

Mode for Carrying Out the Invention

[0026] The present invention is described herein using several definitions set forth below and throughout this application.

[0027] Unless otherwise specified or indicated by the context, the terms "a," "an," and "the" mean "one or more." For example, "compound" should be interpreted to mean "one or more compounds." As used herein, "about," "approximately," "substantially," and "significantly" are understood by those skilled in the art and vary to some extent depending on the context in which they are used. Where the use of a term is not clear to those skilled in the art from the context in which these terms are used, "about" and "approximately" mean plus or minus 10% or less of the particular term, and "substantially" and "significantly" mean plus or minus more than 10% of the particular term.

[0028] As used herein, the terms "include" and "including" have the same meaning as the terms "comprise" and "comprising", and these latter terms are "open" transitional terms that do not limit the scope of the claims to only the recited elements following these transitional terms. The term "consisting of" is included within the term "comprising", but should be construed as a "closed" transitional term that limits the scope of the claims to only the recited elements following this transitional term. The term "consisting essentially of" is included within the term "comprising", but should be construed as a "partially closed" transitional term that allows for additional elements following this transitional term, but only to the extent that those additional elements do not substantially affect the basic and novel characteristics of the scope of the claims.

[0029] As used herein, "subject" can be interchangeable with "patient" or "individual" and means an animal, which can be a human or non-human animal in need of treatment.

[0030] Examples of "subjects in need of treatment" can include subjects having a disease, disorder or condition that responds to treatment with a substituted heterocycle such as a substituted pyrazole, substituted imidazole and substituted triazole of the present disclosure, or a proteolytic targeting chimeric molecule (PROTAC) that targets c-MYC for degradation of c-MYC. For example, "subjects in need of treatment" can include subjects having a cell proliferative disease, disorder or condition such as cancer (e.g., cancers such as multiple myeloma, leukemia, non-small cell lung cancer, colon cancer, central nervous system cancer, melanoma, ovarian cancer, renal cancer, prostate cancer, and breast cancer).

[0031] As used herein, the term "effective amount" shall mean a dosage of a drug that provides a particular pharmacological response for which such treatment is required in a significant number of subjects to whom the drug is administered. The effective amount of a drug administered to a particular subject in a particular instance is not always effective for the treatment of the conditions / diseases described herein, even if such dosage is considered by one of ordinary skill in the art to be a therapeutically effective amount.

[0032] Chemical substance New chemical substances and uses for chemical substances are disclosed herein. Chemical substances can be described using terms known in the art and can be further considered below.

[0033] As used herein, a wavy line

Chem.

[0034] As used herein, the term "alkyl" as contemplated herein refers to herein C1-C 12 -alkyl, C1-C 10 -alkyl, and C1-C6-alkyl, including all straight-chain or branched-chain alkyl radicals of isomers thereof, such as straight-chain or branched-chain groups of 1 to 12, 1 to 10, or 1 to 6 carbon atoms. The number of carbon atoms in an alkyl group can be specified using the Cx-Cy nomenclature, where x and y are integers specifying the number of carbon atoms.

[0035] The term "alkylenyl" refers to a diradical of a straight-chain or branched-chain alkyl group (i.e., a diradical of a straight-chain or branched-chain C1-C6 alkyl group). Exemplary alkylenyl groups include, but are not limited to, -CH2-, -CH2CH2-, -CH2CH2CH2-, -CH(CH3)CH2-, -CH2CH(CH3)CH2-, -CH(CH2CH3)CH2-, etc.

[0036] The term "halo" or "halogen" refers to a radical of -F, -Cl, -Br or -I.

[0037] The term "haloalkyl" refers to an alkyl group substituted with at least one halogen. For example, -CH2F, -CHF2, -CF3, -CH2CF3, -CF2CF3, etc.

[0038] The terms "alkoxy" or "alkoxyl" are recognized in the art and refer to the alkyl group as defined above to which an oxygen radical is attached. Representative alkoxy groups include methoxy, ethoxy, tert-butoxy, etc.

[0039] The term "haloalkoxy" refers to an alkoxy group as defined above substituted with at least one halogen.

[0040] The term "cycloalkyl" as used herein refers to, for example, a monovalent saturated cyclic, bicyclic, or bridged cyclic (e.g., adamantyl) hydrocarbon group of 3 to 12, 3 to 8, 4 to 8, or 4 to 6 carbons, called "C 4~8 -cycloalkyl". The number of carbon atoms in the cycloalkyl group can be specified using the Cx-Cy nomenclature, where x and y are integers specifying the number of carbon atoms. Unless otherwise specified, the cycloalkyl group is optionally substituted at one or more ring positions with, for example, alkanoyl, alkoxy, alkyl, haloalkyl, alkenyl, alkynyl, amide or carboxamide (or amidecarboxyl), amidino, amino, aryl, arylalkyl, azide, carbamate, carbonate, carboxy, cyano, cycloalkyl, ester, ether, formyl, halo, haloalkyl, heteroaryl, heterocyclyl, hydroxyl, imino, ketone, nitro, phosphate, phosphonato, phosphinato, sulfate, sulfide, sulfonamide, sulfonyl or thiocarbonyl. In certain embodiments, the cycloalkyl group is unsubstituted, i.e., non-substituted.

[0041] The term "cycloheteroalkyl" refers to a monovalent saturated cyclic, bicyclic or bridged cyclic hydrocarbon group of 3 to 12, 3 to 8, 4 to 8 or 4 to 6 carbons in which at least one carbon of the cycloalkane is replaced by a heteroatom such as N, O and / or S.

[0042] As used herein, the term "cycloheteroalkenyl" refers to the diradical of a cycloheteroalkyl group as defined above.

[0043] The term "cycloalkenyl" refers to the diradical of a cycloalkyl group as defined above. Exemplary cycloalkenyl groups include,

Chemical formula

[0044] The term "aryl" is recognized in the art and refers to, for example, a carbocyclic aromatic group containing 5 to 12, 6 to 10, or 5 to 8 carbons. The number of carbon atoms in an aryl group can be specified using the Cx-Cy nomenclature, where x and y are integers specifying the number of carbon atoms. Representative aryl groups include phenyl, naphthyl, anthracenyl, and the like. The term "aryl" includes polycyclic ring systems "condensed rings" having two or more carbocyclic rings in which two or more carbons are common to two adjacent rings, with at least one of the rings being aromatic, and for example, one or more of the other ring(s) can be cycloalkyl, cycloalkenyl, cycloalkynyl, and / or aryl. Unless otherwise specified, the aromatic ring can be substituted at one or more ring positions with, for example, halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxyl, amino, nitro, sulfhydryl, imino, amide or carboxamide (or amide carboxyl), carboxylic acid, -C(O)alkyl, -CO2alkyl, carbonyl, carboxyl, alkylthio, sulfonyl, sulfonamide, sulfonamide, ketone, aldehyde, ester, heterocyclyl, aryl or heteroaryl moieties, -CF3, -CN, etc. In certain embodiments, the aromatic ring is substituted at one or more ring positions with halogen, alkyl, hydroxyl, or alkoxyl. In certain other embodiments, the aromatic ring is unsubstituted, i.e., non-substituted. In certain embodiments, the aryl group has a 6- to 10-membered ring structure.

[0045] As used herein, the term "arylenyl" refers to the diradical of an aryl group as defined above.

[0046] The term "alkylaryl" refers to a monoradical having an alkenyl group attached to an aryl group.

[0047] The term "aryloxy" is recognized in the art and refers to an aryl group as defined above having an oxygen radical attached thereto. Representative aryloxy groups include phenoxy and the like.

[0048] The term "alkylaryloxy" refers to a monoradical having an alkenyl group attached to an aryloxy group.

[0049] As used herein, the term "heteroaryl" refers to monocyclic heteroaryl and bicyclic heteroaryl. Monocyclic heteroaryl is a 5-membered or 6-membered ring. The 5-membered ring contains two double bonds. The 5-membered ring contains at least one heteroatom which is O, S and / or nitrogen; or may contain one, two, three or four nitrogen atoms, and optionally one oxygen atom or one sulfur atom. The 6-membered ring contains three double bonds and one, two, three or four nitrogen, oxygen and / or sulfur atoms. Representative examples of monocyclic heteroaryl include, but are not limited to, furanyl, imidazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, 1,3-oxazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, pyrazolyl, pyrrolyl, tetrazolyl, thiadiazolyl, 1,3-thiazolyl, thienyl, triazolyl and triazinyl. Bicyclic heteroaryl consists of monocyclic heteroaryl fused to phenyl, or monocyclic heteroaryl fused to monocyclic cycloalkyl, or monocyclic heteroaryl fused to monocyclic cycloalkenyl, or monocyclic heteroaryl fused to monocyclic heteroaryl, or monocyclic heteroaryl fused to monocyclic heterocycle. Representative examples of bicyclic heteroaryl include, but are not limited to, benzofuranyl, benzothienyl, benzoxazolyl, benzimidazolyl, benzoxadiazolyl, phthalazinyl, 2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl, 6,7-dihydro-pyrazolo[1,5-a]pyrazin-5(4H)-yl, 6,7-dihydro-1,3-benzothiazolyl, imidazo[1,2-a]pyridinyl, indazolyl, indolyl, isoindolyl, isoquinolinyl, naphthyridinyl, pyridoimidazolyl, quinolinyl, 2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl, thiazolo[5,4-b]pyridin-2-yl, thiazolo[5,4-d]pyrimidin-2-yl and 5,6,7,8-tetrahydroquinolin-5-yl. Monocyclic and bicyclic heteroaryl containing exemplary rings are optionally substituted unless otherwise specified.The monocyclic and bicyclic heteroaryls are connected to the parent molecular moiety via any replaceable carbon atom or any replaceable nitrogen atom contained within the ring system. The nitrogen atoms in the heteroaryl ring may be optionally oxidized and may be optionally quaternized.

[0050] The term "heteraryloxy" refers to a heteroaryl group as defined above having an oxygen radical attached thereto.

[0051] As used herein, the term "heterarylenyl" refers to a diradical of a heteroaryl group as defined above.

[0052] The terms "heterocyclyl" and "heterocyclic group" are recognized in the art and refer to a saturated, partially unsaturated, or aromatic 3- to 10-membered ring structure, or a 3- to 7-membered ring, the ring structure of which contains 1 to 4 heteroatoms such as nitrogen, oxygen, and sulfur. The number of ring atoms of a heterocyclyl group can be specified using the "x- to y-membered" nomenclature, where x and y are integers specifying the number of ring atoms. For example, a 3- to 7-membered heterocyclyl group refers to a saturated or partially unsaturated 3- to 7-membered ring structure containing 1 to 4 heteroatoms such as nitrogen, oxygen, and sulfur. The specification indicates that the heterocyclic ring contains a total of 3 to 7 ring atoms, including any heteroatom occupying a ring atom position.

[0053] The terms "amine" and "amino" are recognized in the art and refer to both unsubstituted and substituted amines (e.g., mono-substituted amines or di-substituted amines), the substituents of which can include, for example, alkyl, cycloalkyl, heterocyclyl, alkenyl, and aryl.

[0054] "Ether" is two hydrocarbons covalently linked by oxygen. Thus, the alkyl substituent that makes an alkyl into an ether is an alkoxyl such that it can be represented by one of -O-alkyl, -O-alkenyl, -O-alkynyl, etc. or is similar to such an alkoxyl.

[0055] As used herein, the term "carbonyl" refers to the radical -C(O)-.

[0056] The term "oxo" refers to a divalent oxygen atom

Chem.

[0057] As used herein, the term "carboxamide" refers to the radical -C(O)NRR', where R and R' may be the same or different. R and R' can be, for example, independently hydrogen, alkyl, aryl, arylalkyl, cycloalkyl, formyl, haloalkyl, heteroaryl or heterocyclyl.

[0058] As used herein, the term "carboxy" or "carboxyl" refers to the radical -COOH or its corresponding salt, such as -COONa and the like.

[0059] 1 As used herein, the term "amide" or "amid" or "amidyl" refers to a radical in the form of -R 2 C(O)N(R 1 )-, -R 2 C(O)N(R 3 )R 2 -, -C(O)NR 3 R 1 、R 2 and R 3 are, for example, each independently hydrogen, alkyl, alkoxy, alkenyl, alkynyl, amide, amino, aryl, arylalkyl, carbamate, cycloalkyl, ester, ether, formyl, halogen, haloalkyl, heteroaryl, heterocyclyl, hydrogen, hydroxyl, ketone, or nitro.

[0060] ​As used herein, the term "alkenyl" refers to an unsaturated straight-chain or branched hydrocarbon having at least one carbon-carbon double bond, such as C2-C 12 -alkenyl, C2-C 10 -alkenyl, and straight-chain or branched groups of 2 to 12, 2 to 10, or 2 to 6 carbon atoms referred to herein as C2-C6-alkenyl.

[0061] As used herein, the term "alkynyl" refers to an unsaturated straight-chain or branched hydrocarbon having at least one carbon-carbon triple bond, such as C2-C 12 -alkynyl, C2-C 10 -alkynyl, and straight-chain or branched groups of 2 to 12, 2 to 10, or 2 to 6 carbon atoms referred to herein as C2-C6-alkynyl.

[0062] As used herein, the term "benzyl" refers to the group -C6H4-CH2- (i.e., [Chemical formula] ).

[0063] As used herein, the term "benzylyl" refers to the diradical of the benzyl group defined above.

[0064] The term "cyano" refers to the substituent "-CN".

[0065] The term "hydroxyl" refers to the substituent "-OH".

[0066] The compounds and molecules (e.g., PROTACs) of the present disclosure may contain one or more chiral centers and / or double bonds and, accordingly, may exist as stereoisomers, such as geometric isomers, enantiomers or diastereomers. As used herein, the term “stereoisomers” consists of all geometric isomers, enantiomers or diastereomers. These compounds and molecules may be designated by the symbols “R” or “S” or “+” or “-” depending on the arrangement of substituents around the stereogenic carbon atoms and / or the observed optical rotation. The present invention encompasses the various stereoisomers of these compounds and molecules and mixtures thereof. Stereoisomers include enantiomers and diastereomers. A mixture of enantiomers or diastereomers may be designated as (±) in nomenclature, but one of ordinary skill in the art will recognize that the structure may imply a chiral center. It is understood that a chemical structure, e.g., a graphical representation of a general chemical structure, includes all stereoisomeric forms of the designated compounds and molecules unless otherwise indicated. Compositions that are enantiopure, consisting essentially of, or consisting of, are also contemplated herein, and such compositions may contain, consist essentially of, or consist of at least about 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% of a single enantiomer of a given compound (e.g., at least about 99% of the R enantiomer of a given compound).

[0067] The formulas of the compounds and molecules disclosed herein should be interpreted to include all possible stereoisomers, enantiomers or epimers of the compounds and molecules unless the formula indicates a specific stereoisomer, enantiomer or epimer. The formulas of the compounds and molecules disclosed herein should be interpreted to include salts, esters, amides, or solvates of the compounds and molecules.

[0068] Substituted Heterocycles as MYC Inhibitors A substituted heterocyclic ring is disclosed herein. The disclosed heterocyclic ring has been shown to inhibit the biological activity of c-MYC. The disclosed substituted heterocyclic ring may include a substituted pyrazole, a substituted imidazole, and a substituted triazole.

[0069] In some embodiments, the disclosed substituted heterocyclic ring has the formula I:

Chemical formula

[0070] In some embodiments, the disclosed substituted heterocycle can have the structure of Formula I, wherein R 1 is hydrogen, alkyl, aryl, benzyl, and R 1 is one or more alkyl, alkoxy, aryl, halo, -(CH2)-NH-R 9 , or -OR11 is optionally substituted at one or more positions with R 2 is hydrogen or chloro, R 3 is hydroxyl or -OC(O)Me, R 4 is hydrogen or halo, R 5 is hydrogen or halo, R 6 is hydrogen, aryl, benzyl, and optionally, R 6 is substituted at one or more positions with haloalkyl, halo, and cyano, R 8 is cyano, amino, haloalkyl or amide, R 14 is aryl optionally substituted at one or more ring positions with one or more haloalkyl or halo, The remaining substituents of the compound of formula I are as defined herein.

[0071] In some embodiments, the compound is not 4'-chloro-6-((4-chlorobenzyl)oxy)-3-(1-methyl-3-(trifluoromethyl)-1H-pyrazol-5-yl)-3'-(trifluoromethyl)-[1,1'-biphenyl]-2-ol, or a compound disclosed in U.S. Patent Application Publication No. 2017 / 0253581, published on September 7, 2017, U.S. Patent Application Publication No. 2019 / 0062281, published on February 28, 2019, U.S. Patent Application Publication No. 2020 / 0390894, published on December 17, 2020, or U.S. Patent Application Publication No. 2020 / 0392116, published on December 17, 2020.

[0072] In some embodiments, the disclosed substituted heterocycle can have formula I(a), and the definitions of the substituents are the same as those of formula I:

Chemical formula

[0073] In some embodiments, the disclosed substituted heterocycle may include a substituted pyrazole where Y is N and W is CCF3.

[0074] In some embodiments, the disclosed substituted heterocycle may have R as hydroxyl. 3

[0075] In some embodiments, the disclosed substituted heterocycle may have r = 0 and R may be phenyl substituted with chloro and trifluoromethyl. 6

[0076] In some embodiments, the disclosed substituted heterocycle may have R as hydrogen. 2

[0077] In some embodiments, the disclosed substituted heterocycle

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0078] ​​​Proteolysis-targeting chimeric molecule (PROTAC) that induces the degradation of c-MYC protein Also disclosed herein are proteolysis-targeting chimeric molecules (PROTACs) that induce the degradation of c-MYC protein. In some embodiments, the disclosed molecules have the formula: M c-MYC -L-M E3 or alternatively M E3 -L-M c-MYC and may be described as having, wherein M c-MYC is a moiety that binds to c-MYC, L is a bond or linker that covalently attaches M MYC and M E3 and M E3 is a moiety that binds to an E3 ubiquitin ligase. Compounds that bind to c-MYC are disclosed in the prior art and may include, but are not limited to, the compounds disclosed in U.S. Patent Application Publication No. 2019 / 0062281, published on February 28, 2019, the contents of which are incorporated herein by reference in their entirety.

[0079] In some embodiments of the disclosed PROTACs, the PROTAC has the formula: M c-MYC -L-M E3 (wherein M c-MYC is a moiety that binds to c-MYC, L is a bond or linker that covalently attaches M MYC and M E3 and M E3 is a moiety that binds to an E3 ubiquitin ligase).

[0080] In some embodiments of the disclosed PROTACs, the PROTAC has Formula II:

Chemical formula

Chemical formula

[0081] In some embodiments, the PROTAC is of formula II(a):

Chemical formula

[0082] In some embodiments of the disclosed PROTACs, the PROTAC has the structure of formula II(a), where R 3 is hydroxyl and R 4 is phenyl substituted with trifluoromethyl and chloro.

[0083] In some embodiments of the disclosed PROTACs, the PROTAC has the structure of formula II(a) where X is -N-C(O)R 6 and R 6 is phenyl substituted with trifluoromethyl and chloro.

[0084] In some embodiments of the disclosed PROTACs, the PROTAC has a structure of Formula II(a) where X is O and Y is propylenyl or benzylenyl.

[0085] In some embodiments of the disclosed PROTACs, the PROTAC

Chemical formula

Chemical formula

[0086] In some embodiments of the disclosed PROTACs, the PROTAC is selected from the following:

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0087] The formulas of the compounds disclosed herein should be interpreted to include all possible stereoisomers, enantiomers, or epimers of the compounds, unless the formula indicates a specific stereoisomer, enantiomer, or epimer. The formulas of the compounds disclosed herein should be interpreted to include salts, esters, amides, or solvates of the compounds.

[0088] The disclosed compounds may exhibit one or more biological activities. The disclosed compounds may inhibit the binding of the Myc / Max complex to DNA (e.g., in a DNA gel shift assay). In some embodiments, the disclosed compounds inhibit the binding of the Myc / Max complex to DNA by at least 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98% or 99% at a concentration of about 100 μM, 50 μM, 10 μM, 1 μM, 0.1 μM, 0.05 μM, 0.01 μM, 0.005 μM, 0.001 μM or less. The disclosed compounds may not cause significant DNA damage (e.g., in an rH2AX staining assay, at a concentration of about 0.001 μM, 0.005 μM, 0.01 μM, 0.1 μM, 1.0 μM, 10 μM, 100 μM or greater). The disclosed compounds may inhibit the growth of cells expressing c-Myc (preferably by at least 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98% or 99% at a concentration of about 100 μM, 50 μM, 10 μM, 1 μM, 0.1 μM, 0.05 μM, 0.01 μM, 0.005 μM, 0.001 μM or less). The disclosed compounds may not inhibit the growth of cells not expressing c-Myc (preferably 50%, 40%, 30%, 20%, 10%, 5%, 4%, 3%, 2% or less at concentrations of about 0.001 μM, 0.005 μM, 0.01 μM, 0.5 μM, 0.1 μM, 1.0 μM, 10 μM and greater than 100 μM). Concentration ranges, e.g., concentration ranges bounded by endpoint concentrations selected from 0.001 μM, 0.005 μM, 0.01 μM, 0.5 μM, 0.1 μM, 1.0 μM, 10 μM and 100 μM are also contemplated herein.

[0089] The disclosed compounds may be effective in inhibiting the cell proliferation of cancer cells, including cancer cells that express c-MYC and whose proliferation is inhibited by inhibiting the biological activity of c-MYC. The disclosed compounds include multiple myeloma cells such as MM.1S cells; leukemia cells such as CCRF-CEM, HL-60 (TB), MOLT-4, RPMI-8226, and SR; non-small lung cancer cells such as A549 / ATCC, EKVX, HOP-62, HOP-92, NCI-H226, NCI-H23, NCI-H322M, NCI-H460, and NCI-H522; colon cancer cells such as COLO 205, HCC-2998, HCT-116, HCT-15, HT29, KM12, and SW-620; CNS: SF-268, SF-295, SF-539, SNB-19, SNB-75, and U251; melanoma cancer cells such as LOX IMVI, MALME-3M, M14, MDA-MB-435, SK-MEL-2, SK-MEL-28, SK-MEL-5, UACC-257, and UACC-62; ovarian cancer cells such as IGR-OV1, OVCAR-3, OVCAR-4, OVCAR-5, OVCAR-8, NCI / ADR-RES, and SK-OV-3; renal cancer cells such as 786-0, A498, ACHN, CAKI-1, RXF 393, SN12C, TK-10, and UO-31; prostate cancer cells such as DU-145 and PC-3; and breast cancer cells such as MCF7, MDA-MB-231 / ATCC, MDA-MB-468, HS 578T, BT-549, and T-47D, and may be effective in inhibiting the cell proliferation of one or more types of cancer cells.

[0090] The cell proliferation and its inhibition by the compounds of the present disclosure can be evaluated by cell viability methods disclosed in the art, including colorimetric assays that utilize dyes such as MTT, XTT, and MTS to evaluate cell viability. Preferably, the disclosed compounds have an IC 50 of about 10 μM, 5 μM, 1 μM, 0.5 μM, 0.01 μM, 0.005 μM, 0.001 μM, or less in the selected assay.

[0091] The disclosed compounds can be formulated as anti-cancer therapeutics, including those for hematological malignancies, breast, lung, pancreatic, and prostate cancers. The disclosed compounds can also be formulated as anti-inflammatory therapeutics.

[0092] The compounds utilized in the methods disclosed herein can be formulated as pharmaceutical compositions comprising (a) a therapeutically effective amount of one or more of the compounds disclosed herein and (b) one or more pharmaceutically acceptable carriers, excipients, or diluents. The pharmaceutical composition can contain the compound in the range of about 0.1 to 2000 mg (preferably about 0.5 to 500 mg, more preferably about 1 to 100 mg). The pharmaceutical composition can be administered to provide the compound at a daily dose of about 0.1 to 100 mg / kg body weight (preferably about 0.5 to 20 mg / kg body weight, more preferably about 0.1 to 10 mg / kg body weight). In some embodiments, after the pharmaceutical composition is administered to a subject (e.g., about 1, 2, 3, 4, 5, or 6 hours after administration), the concentration of the compound at the site of action can be within a concentration range bounded by endpoints selected from 0.001 μM, 0.005 μM, 0.01 μM, 0.5 μM, 0.1 μM, 1.0 μM, 10 μM, and 100 μM (e.g., 0.1 μM to 1.0 μM).

[0093] The disclosed compounds and pharmaceutical compositions containing the disclosed compounds can be administered in methods of treating a subject in need thereof. For example, in a method of treatment, the subject in need thereof can include a subject having a cell proliferative disease, disorder, or condition such as cancer (e.g., cancers such as multiple myeloma, leukemia, non-small cell lung cancer, colon cancer, central nervous system cancer, melanoma, ovarian cancer, renal cancer, prostate cancer, and breast cancer).

[0094] In some embodiments of the disclosed treatment methods, a subject may be administered a low dose of a compound, such as 1.25 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg, 12.5 mg, 15 mg, 17.5 mg, 20 mg, 22.5 mg, 25 mg, 27.5 mg, 30 mg, 32.5 mg, 35 mg, 37.5 mg, 40 mg, 42.5 mg, 45 mg, 47.5 mg, 50 mg, 52.5 mg, 55 mg, 57.5 mg, 60 mg, 62.5 mg, 65 mg, 67.5 mg, 70 mg, 72.5 mg, 75 mg, 77.5 mg, 80 mg, 82.5 mg, 85 mg, 87.5 mg, 90 mg, 100 mg, 200 mg, 500 mg, 1000 mg or 2000 mg, once a day, twice a day, three times a day, four times a day, once a week, twice a week or three times a week, to treat a disease or disorder in the subject. In some embodiments, a subject may be administered a high dose of a compound, such as 1.25 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg, 12.5 mg, 15 mg, 17.5 mg, 20 mg, 22.5 mg, 25 mg, 27.5 mg, 30 mg, 32.5 mg, 35 mg, 37.5 mg, 40 mg, 42.5 mg, 45 mg, 47.5 mg, 50 mg, 52.5 mg, 55 mg, 57.5 mg, 60 mg, 62.5 mg, 65 mg, 67.5 mg, 70 mg, 72.5 mg, 75 mg, 77.5 mg, 80 mg, 82.5 mg, 85 mg, 87.5 mg, 90 mg, 100 mg, 200 mg, 500 mg, 1000 mg or 2000 mg, once a day, twice a day, three times a day, four times a day, once a week, twice a week or three times a week, to treat the subject's disease or disorder. The minimum and / or maximum dose of the compound may include a dose within a dose range having any of these disclosed doses (e.g., 2.5 mg to 200 mg) as an endpoint.

[0095] In some embodiments, the minimum dosage level of the compound to achieve treatment in the disclosed treatment methods can be at least about 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1200, 1400, 1600, 1800, 1900, 2000, 3000, 4000, 5000, 6000, 7000, 8000, 9000, 10000, 15000, or 20000 ng / kg body weight of the subject. In some embodiments, the maximum dosage level of the compound to achieve treatment in the disclosed treatment methods cannot exceed about 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1200, 1400, 1600, 1800, 1900, 2000, 3000, 4000, 5000, 6000, 7000, 8000, 9000, 10000, 15000, or 20000 ng / kg body weight of the subject. The minimum and / or maximum dosage levels of the compound to achieve treatment in the disclosed treatment methods can include dosage levels that fall within a range having any of these disclosed dosage levels as endpoints (e.g., 500 - 2000 ng per kg body weight of the subject).

[0096] The compounds utilized in the methods disclosed herein can be formulated as a pharmaceutical composition in a solid dosage form, although any pharmaceutically acceptable dosage form can be utilized. Exemplary solid dosage forms include, but are not limited to, tablets, capsules, sachets, lozenges, powders, pills, or granules, and the solid dosage form can be, for example, an immediate release dosage form, a controlled release dosage form, a lyophilized dosage form, a delayed release dosage form, a sustained release dosage form, a pulsatile release dosage form, a mixed immediate release and controlled release dosage form, or a combination thereof.

[0097] The compounds utilized in the methods disclosed herein can be formulated as pharmaceutical compositions containing a carrier. For example, the carrier can be selected from the group consisting of proteins, carbohydrates, sugars, talc, magnesium stearate, cellulose, calcium carbonate, and starch gelatin paste.

[0098] The compounds utilized in the methods disclosed herein can be formulated as pharmaceutical compositions containing one or more binders, fillers, lubricants, suspending agents, sweeteners, flavoring agents, preservatives, buffers, wetting agents, disintegrants, and foaming agents. Examples of fillers can include lactose monohydrate, lactose anhydrous, and various starches. Examples of binders are various celluloses and cross-linked polyvinylpyrrolidone, microcrystalline cellulose such as Avicel® PH101 and Avicel® PH102, microcrystalline cellulose, and silicified microcrystalline cellulose (ProSolv SMCC™). Suitable lubricants containing agents that act on the flowability of the compressed powder can include colloidal silicon dioxide such as Aerosil® 200, talc, stearic acid, magnesium stearate, calcium stearate, and silica gel. Examples of sweeteners can include any natural or artificial sweeteners such as sucrose, xylitol, sodium saccharin, cyclamate, aspartame, and acesulfame. Examples of flavoring agents are Magnasweet® (a trademark of MAFCO), bubble gum flavor, fruit flavor, etc. Examples of preservatives can include potassium sorbate, methyl paraben, propyl paraben, benzoic acid and its salts, other esters of p-hydroxybenzoic acid such as butyl paraben, alcohols such as ethyl or benzyl alcohol, phenolic compounds such as phenol, or quaternary compounds such as benzalkonium chloride.

[0099] Suitable diluents may include pharmaceutically acceptable inert fillers such as microcrystalline cellulose, lactose, dibasic calcium phosphate, saccharides, and mixtures of any of the foregoing. Examples of diluents include microcrystalline cellulose such as Avicel® PH101 and Avicel® PH102; lactose such as lactose monohydrate, lactose anhydrous, Pharmatose® DCL21; dibasic calcium phosphate such as Emcompress®; mannitol; starch; sorbitol, sucrose; and glucose.

[0100] Suitable disintegrants may include lightly cross-linked polyvinylpyrrolidone, corn starch, potato starch, corn starch, and modified starch, croscarmellose sodium, crospovidone, sodium starch glycolate, and mixtures thereof.

[0101] Examples of foaming agents are foaming couples such as organic acids and carbonates or bicarbonates. Suitable organic acids include, for example, citric acid, tartaric acid, malic acid, fumaric acid, adipic acid, succinic acid and alginic acid and anhydrides and salts. Suitable carbonates and bicarbonates include, for example, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, magnesium carbonate, sodium glycine carbonate, L-lysine carbonate and arginine carbonate. Alternatively, only the sodium bicarbonate component of the foaming pair may be present.

[0102] The compounds utilized in the methods disclosed herein can be formulated as pharmaceutical compositions for delivery via any suitable route. For example, the pharmaceutical compositions can be administered via oral, intravenous, intramuscular, subcutaneous, topical and pulmonary routes. Examples of pharmaceutical compositions for oral administration include capsules, syrups, concentrates, powders and granules.

[0103] The compounds utilized in the methods disclosed herein can be administered in conventional dosage forms prepared by combining the active ingredient with a standard pharmaceutical carrier or diluent according to conventional procedures well known in the art. These procedures can include mixing, granulating, and compressing or dissolving the ingredients as appropriate for the desired preparation.

[0104] The pharmaceutical composition containing the compound may be adapted for administration by any suitable route, such as oral (including buccal or sublingual), rectal, nasal, topical (including buccal, sublingual or transdermal), vaginal or parenteral (including subcutaneous, intramuscular, intravenous or intradermal) routes. Such formulations can be prepared by any method known in the art of pharmacy, for example by associating the active ingredient with a carrier(s) or excipient(s).

[0105] Pharmaceutical compositions adapted for oral administration may be provided as individual units, such as capsules or tablets; powders or granules; solutions or suspensions in aqueous or non-aqueous liquids; edible forms or whips; or oil-in-water or water-in-oil liquid emulsions.

[0106] Pharmaceutical compositions adapted for transdermal administration may be provided as individual patches intended to remain in intimate contact with the recipient's epidermis for an extended period of time. For example, the active ingredient can be delivered from the patch by iontophoresis.

[0107] Pharmaceutical compositions adapted for topical administration may be formulated as ointments, creams, suspensions, lotions, powders, solutions, pastes, gels, impregnated dressings, sprays, aerosols or oils and may contain suitable conventional additives such as preservatives, solvents to assist drug penetration and skin softening agents in ointments and creams.

[0108] For application to the eye or other external tissues, such as the mouth and skin, the pharmaceutical composition is preferably applied as a topical ointment or cream. When formulated as an ointment, the compound can be used with either a paraffinic ointment base or a water-miscible ointment base. Alternatively, the compound may be formulated into a cream containing an oil-in-water cream base or a water-in-oil base. Pharmaceutical compositions suitable for topical administration to the eye include eye drops in which the active ingredient is dissolved or suspended in a suitable carrier, particularly an aqueous solvent.

[0109] Pharmaceutical compositions suitable for nasal administration in which the carrier is solid include coarse powders having a particle size (e.g., in the range of 20 to 500 microns) for administration in a manner in which snuff is taken (i.e., by rapid inhalation through the nasal passages from a container of powder held close to the nose). When the carrier is liquid, suitable formulations for administration as a nasal spray or nasal drops include aqueous solutions or oily solutions of the active ingredient.

[0110] Pharmaceutical compositions suitable for parenteral administration include aqueous and non-aqueous isotonic sterile injection solutions which may contain antioxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient, and aqueous and non-aqueous sterile suspensions which may contain suspending and thickening agents. The formulations can be presented in unit-dose or multi-dose containers, for example, sealed ampoules and vials, and can be stored in a freeze-dried (lyophilized) state that requires only the addition of a sterile liquid carrier, for example, water, immediately prior to use for injection. Immediate injection solutions and suspensions can be prepared from sterile powders, granules, and tablets.

[0111] Tablets and capsules for oral administration can be in unit dosage presentation form and can contain conventional excipients such as binders, for example, syrup, acacia, gelatin, sorbitol, tragacanth or polyvinylpyrrolidone; fillers, for example, lactose, sugar, corn starch, calcium phosphate, sorbitol or glycine; tablet lubricants, for example, magnesium stearate, talc, polyethylene glycol or silica; disintegrants, for example, potato starch; or acceptable wetting agents such as sodium lauryl sulfate. Tablets can be coated according to methods well known in ordinary pharmaceutical practice. Oral liquid preparations can be in the form of, for example, aqueous or oily suspensions, solutions, emulsions, syrups or elixirs, or can be provided as dry products for reconstitution with water or other suitable vehicles before use. Such liquid preparations can contain suspending agents, for example, sorbitol, methylcellulose, glucose syrup, gelatin, hydroxyethylcellulose, carboxymethylcellulose, aluminum stearate gel or hydrogenated edible fats, emulsifying agents, for example, lecithin, sorbitan monooleate or acacia; non-aqueous vehicles (which can include edible oils), for example, almond oil, glycerin, propylene glycol or oily esters such as ethyl alcohol; preservatives, for example, methyl or propyl p-hydroxybenzoate or sorbic acid, and optionally conventional additives such as conventional flavoring or coloring agents.

[0112] The disclosed compounds or pharmaceutical compositions containing the disclosed compounds can be administered in a method of treatment. For example, the disclosed compounds or pharmaceutical compositions containing the disclosed compounds can be administered in a method of treating cell proliferative diseases and disorders. Cell proliferative diseases and disorders treatable by the disclosed methods can include, but are not limited to, cancers selected from the group consisting of multiple myeloma, leukemia, non-small cell lung cancer, colon cancer, cancers of the central nervous system, melanoma, ovarian cancer, renal cancer, prostate cancer, and breast cancer.

[0113] Optionally, the disclosed compounds or pharmaceutical compositions comprising the disclosed compounds can be administered, optionally in combination with additional therapeutic agents, for treating cell proliferative diseases and disorders. In some embodiments of the disclosed methods, one or more additional therapeutic agents are administered with the disclosed compounds or pharmaceutical compositions comprising the disclosed compounds, and the additional therapeutic agents are administered before, simultaneously with, or after administration of the disclosed compounds or pharmaceutical compositions comprising the disclosed compounds. In some embodiments, the disclosed pharmaceutical compositions comprise the disclosed compounds and are formulated to further comprise one or more additional therapeutic agents, such as one or more additional therapeutic agents for treating cell proliferative diseases and disorders.

[0114] In some embodiments, additional therapeutic agents can include, but are not limited to, therapeutics for treating leukemia and lymphoma, such as acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), chronic myeloid leukemia (CML), and non-Hodgkin lymphoma.

[0115] In some embodiments, additional therapeutic agents can include, but are not limited to, antimetabolite antineoplastic agents that inhibit DNA synthesis. Suitable antimetabolite antineoplastic agents that inhibit DNA synthesis can include, but are not limited to, nucleoside and / or nucleotide derivatives. Suitable nucleoside and / or nucleotide derivatives can include, but are not limited to, cytosine arabinoside (ara-C) (also known as cytarabine).

Example

[0116] The following examples are illustrative and are not intended to limit the scope of the claimed subject matter.

[0117] Example 1 Synthesis of A4BC1R1 (NUCC-0226301), A4BC1R3 (NUCC-0226276), A4BC1R4 (NUCC-0226302), A4BC1R5 (NUCC-0226277) and A4BC1_2Ts (NUCC-0226263) Synthesis scheme

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

[0118] LCMS method: Method 1: 5_95AB_6 min - 220 - 254 - ELSD LC / MS (Gradient: 5% B for 0.40 min, 5 - 95% B from 0.40 to 3.00 min, hold at 95% B for 1.00 min, then 95 - 5% B in 0.01 min; flow rate: 1.0 mL / min. Mobile phase A: 0.037% trifluoroacetic acid in water; mobile phase B: 0.018% trifluoroacetic acid in acetonitrile. Column used for chromatography: Kinetex C18 50×2.1 mm column (5 μm particles). Detection methods: diode array (DAD) and evaporative light scattering (ELSD) detection and positive electrospray ionization. MS range: 100 - 1000.)

[0119] Method 2: 5_95CD_6 min - 220 - 254 - ELSD LC / MS (Gradient: 5% B for 0.40 min, 5 - 95% B from 0.40 to 3.40 min, hold at 95% B for 0.45 min, then 95 - 5% B in 0.01 min; flow rate: 0.8 mL / min. Mobile phase A: H2O + 10 mM NH4HCO3; mobile phase B: acetonitrile. Column used for chromatography: Xbridge Shield RP18 2.1×50 mm column (5 μm particles). Detection methods: diode array (DAD) and evaporative light scattering (ELSD) detection and positive electrospray ionization. MS range: 100 - 1000.)

[0120] Method 3: 5 - 95AB_2 min For LC / MS, the column used in chromatography was Kinetex 5μm EVO C18 100A 2.1×30mm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in HPLC-grade acetonitrile. The gradient was 5 - 95% B in 1.50 minutes, 5% B for 0.01 minute, 5 - 95% B (0.01 - 0.70 minute), 95% B for 0.46 minute, 95 - 5% B (1.61 - 1.50 minute), and held at 5% B for 0.11 minute. The flow rate was 1.5 mL / min.

[0121] Method 4: 5_95CD_6 minutes_MS1500 - 220 - 254 - ELSD For LC / MS, the gradient was 5% B in 0.40 minute, 5 - 95% B from 0.40 to 3.40 minutes, held at 95% B for 0.45 minute, then 95 - 5% B in 0.01 minute, and the flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3, and mobile phase B was acetonitrile. The column used in chromatography was an Xbridge C18 2.1×50mm column (5μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1500.

[0122] Method 5: 5_95AB_6 minutes_MS1500 - 220 - 254 - ELSD LC / MS (Gradient: 5% B from 0.40 min, 5 - 95% B from 0.40 - 3.00 min, hold at 95% B for 1.00 min, then 95 - 5% B in 0.01 min, flow rate was 1.0 mL / min. Mobile phase A was 0.04% trifluoroacetic acid in water and mobile phase B was 0.02% trifluoroacetic acid in acetonitrile. The column used for chromatography was Luna C18 50×2.0 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1500.)

[0123] Experiment for the largest scale execution: General procedure for the preparation of Cpd 2 - ET37412 - 62

[0124] [Chemical formula] To a solution of DHP (2.3 g, 27.29 mmol, 1.7 equiv) in DCM (20 mL), Cpd 1 (2 g, 16.06 mmol, 1 equiv) and TsOH (276 mg, 1.61 mmol, 0.1 equiv) were added at once under N2 at 25 °C. The mixture was stirred at 25 °C for 12 h. TLC indicated the completion of the reaction. The reaction mixture was concentrated to give a residue, which was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100 - 200 mesh silica gel, petroleum ether / ethyl acetate = 1 / 0, 0 / 1) to give Cpd 2 (2 g, yield 59.69%) as a colorless oil.

[0125] 1 1H NMR (ET37412 - 62 - 1, 400 MHz, chloroform - d) δ 1.47 - 1.66 (m, 6H), 1.70 - 1.77 (m, 1H), 1.80 - 1.90 (m, 1H), 3.47 - 3.55 (m, 1H), 3.58 - 3.67 (m, 3H), 3.69 - 3.74 (m, 2H), 3.76 - 3.81 (m, 2H), 3.84 - 3.92 (m, 2H), 4.65 (t, J = 3.64 Hz, 1H)

[0126] General procedure for the preparation of Cpd 3-ET37412 [Chemical formula] To a solution of 4-hydroxybenzaldehyde (900 mg, 7.37 mmol, 1 equiv) in DMA (10 mL), Cpd 2 (1.85 g, 8.84 mmol, 1.2 equiv) and K2CO3 (2.04 g, 14.74 mmol, 2 equiv) were added at once under N2 at 25 °C. The mixture was stirred at 90 °C for 12 h. LCMS indicated that the reaction was complete. The reaction mixture was poured into ice water (10 mL). The aqueous phase was extracted with ethyl acetate (10 mL × 2). The combined organic phases were washed with brine (10 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give Cpd 3 (1.5 g, yield 69.15%) as a colorless oil.

[0127] LCMS (ESI+): RT = 0.824 min, m / z 211.0 (M-DHP) +

[0128] 5 - 95 AB_2 min LC / MS (The column used for chromatography was Kinetex 5μm EVO C18 100A 2.1×30 mm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in HPLC grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B for 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B for 0.46 min, 95 - 5% B (1.61 - 1.50 min), and held at 5% B for 0.11 min. The flow rate was 1.5 mL / min.

[0129] General procedure for the preparation of BC1_DHP-ET37412-71

[0130] [Chemical formula] To a solution of Cpd 3 (1.4 g, 23.78 mmol, 1 equiv) in MeOH (14 mL), NaBH4 (269 mg, 7.13 mmol, 1.5 equiv) was added at once at 25 °C under N2. The mixture was stirred at 25 °C for 12 h. The mixture was poured into ice water (10 mL). The aqueous phase was extracted with ethyl acetate (10 mL × 2). The combined organic phases were washed with brine (10 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100 - 200 mesh silica gel, petroleum ether / ethyl acetate = 1 / 0, 0 / 1) to give BC1_DHP (1 g, yield 71%) as a colorless oil.

[0131] 1 H NMR (ET37412 - 71 - 1 400 MHz, chloroform - d) δ 1.48 - 1.65 (m, 5H), 1.70 - 1.78 (m, 1H), 1.79 - 1.91 (m, 1H), 3.48 - 3.57 (m, 1H), 3.62 - 3.69 (m, 1H), 3.77 (t, J = 4.83 Hz, 2H), 3.84 - 3.95 (m, 4H), 4.08 - 4.24 (m, 2H), 4.59 - 4.71 (m, 3H), 6.93 (d, J = 8.56 Hz, 2H), 7.19 - 7.37 (m, 3H)

[0132] General procedure for the preparation of Cpd 7 - ET43259 - 3

Chemical formula

[0133] 11H NMR (ET37412-92-1, 400 MHz, chloroform-d) δ 3.63 - 3.71 (m, 1H), 3.63 - 3.71 (m, 1H), 3.78 (br d, J = 3.79 Hz, 2H), 3.88 - 3.97 (m, 2H), 4.19 - 4.26 (m, 2H), 7.03 (br d, J = 8.44 Hz, 2H), 7.70 - 7.96 (m, 2H), 9.77 - 9.99 (m, 1H)

[0134] General procedure for the preparation of Cpd 8 - ET37412-103 [Chemical formula] To a solution of Cpd 7 (1 g, 4.76 mmol, 1 equiv) in DCM (10 mL), TEA (1.93 g, 19.03 mmol, 4 equiv) and TosCl (1.36 g, 7.14 mmol, 1.5 equiv) were added at once under N2 at 25 °C. The mixture was stirred at 25 °C for 12 h. The residue was poured into ice water (10 mL). The aqueous phase was extracted with DCM (10 mL). The organic phase was washed with brine (5 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give Cpd 8 (1 g, yield 58%) as a colorless oil.

[0135] LCMS (ESI+): RT = 0.931 min, m / z 365.0 (M + 1) +

[0136] 5 - 95 AB_2 min LC / MS (The column used for chromatography was Kinetex 5μm EVO C18 100A 2.1×30 mm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in HPLC grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B for 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B for 0.46 min, 95 - 5% B (1.61 - 1.50 min), and held at 5% B for 0.11 min. The flow rate was 1.5 mL / min.)

[0137] General procedure for the preparation of Cpd 9-ET37412-105

[0138]

Chem.

[0139] 1 H NMR (ET37412-105-1, 400 MHz, chloroform-d) δ 2.43 (s, 3H), 2.66 (dd, J = 6.72, 2.57 Hz, 1H), 3.70 - 3.81 (m, 4H), 4.00 - 4.23 (m, 5H), 4.63 (s, 2H), 6.85 - 6.92 (m, 2H), 7.28 - 7.41 (m, 5H), 7.80 (d, J = 8.31 Hz, 2H)

[0140] General procedure for the preparation of BC1_Pht ET37412-115

Chem.

[0141] 1 H NMR (ET37412-115-1, 400 MHz, chloroform-d) δ 3.79 - 3.87 (m, 5H), 3.89 - 3.95 (m, 2H), 4.03 - 4.09 (m, 2H), 4.59 (s, 2H), 6.77 - 6.83 (m, 2H), 7.22 (br d, J = 7.06 Hz, 2H), 7.64 - 7.74 (m, 2H), 7.78 - 7.87 (m, 2H)

[0142] General procedure for the preparation of A4BC1_2-DHP-ET37412-96

Chemical formula

[0143] LCMS (ESI+): RT = 0.976 min, m / z 779.3 (M + 23) +

[0144] 5 - 95 AB_2 min LC / MS (The column used for chromatography was Kinetex 5μm EVO C18 100A 2.1×30 mm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in HPLC grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B for 0.46 min, 95 - 5% B (1.61 - 1.50 min), and held at 5% B for 0.11 min. The flow rate was 1.5 mL / min.)

[0145] General procedure for the preparation of A4BC1_2 - ET37412 - 106

[0146]

Chemical formula

[0147] General procedure for the preparation of A4BC1_2Ts (NUCC - 0226263) - ET37412 - 104

[0148]

Chemical formula

[0149] LCMS (ESI+): RT = 0.969 min, m / z 827.2 (M+1) +

[0150] 5 - 95AB_2 min LC / MS (The column used for chromatography was Kinetex 5μm EVO C18 100A 2.1×30 mm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in HPLC grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B for 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B for 0.46 min, 95 - 5% B (1.61 - 1.50 min), and held at 5% B for 0.11 min. The flow rate was 1.5 mL / min.

[0151] General procedure for the preparation of A4BC1_2Pht - ET37412 - 117

[0152]

Chemical formula

[0153] General procedure for the preparation of A4BC1_NH2-ET37412-128

[0154] [Chemical formula] A solution of A4BC1_2-Pht (400 mg, 498.68 μmol, 1 equiv) in EtOH (5 mL) was added with NH2NH2·H2O (312.05 mg, 4.99 mmol, 302.96 μL, purity 80%, 10 equiv) under N2 at 25 °C. The mixture was stirred at 25 °C for 1 h. LCMS indicated that the reaction was complete. The mixture was concentrated to obtain a residue. The residue was purified by Prep-HPLC (column: Waters Xbridge BEH C18 100×30 mm×10 μm; mobile phase: [water (10 mM NH4HCO3)-ACN]; B%: 50% - 75%, 8 min) to obtain A4BC1_NH2 (200 mg, 317.47 μmol, yield 63.66%) as a white solid.

[0155] 11H NMR (ET37412-128-1, 400 MHz, chloroform-d) δ 2.96 (br s, 2H), 3.55 - 3.67 (m, 3H), 3.77 - 3.86 (m, 6H), 4.07 - 4.16 (m, 2H), 5.02 (s, 2H), 6.51 - 6.59 (m, 1H), 6.57 (s, 1H), 6.76 (br d, J = 8.68 Hz, 1H), 6.88 (br d, J = 8.44 Hz, 2H), 7.15 (br d, J = 8.44 Hz, 2H), 7.20 (br d, J = 8.56 Hz, 1H), 7.49 - 7.54 (m, 1H), 7.56 - 7.61 (m, 1H), 7.78 (s, 1H)

[0156] General procedure for the preparation of A4BC1R1(NUCC-0226301)-ET37412-130

Chemical formula

[0157] 11H NMR (ET37412-130-11, 400 MHz, chloroform-d) δ 2.08 - 2.15 (m, 1H), 2.66 - 2.98 (m, 3H), 3.51 (t, J = 5.32 Hz, 2H), 3.80 (t, J = 5.38 Hz, 2H), 3.84 (s, 3H), 3.86 - 3.89 (m, 2H), 4.11 - 4.16 (m, 2H), 4.91 (dd, J = 12.04, 5.32 Hz, 1H), 5.02 (s, 2H), 5.16 - 5.27 (m, 1H), 5.16 - 5.27 (m, 1H), 6.57 (s, 1H), 6.78 (d, J = 8.56 Hz, 1H), 6.85 - 6.90 (m, 2H), 6.93 (d, J = 8.56 Hz, 1H), 7.09 (d, J = 6.97 Hz, 1H), 7.15 (d, J = 8.68 Hz, 2H), 7.21 (d, J = 8.56 Hz, 1H), 7.47 (dd, J = 8.50, 7.15 Hz, 1H), 7.51 - 7.55 (m, 1H), 7.58 - 7.62 (m, 1H), 7.78 (d, J = 1.83 Hz, 1H), 8.01 (s, 1H) LCMS (ESI+): RT = 3.602 min, m / z 886.2 (M+1) +

[0158] 5_95 CD_6 min_MS1500 - 220 - 254 - ELSD: LC / MS (Gradient was 5% B from 0.40 min, 5 - 95% B from 0.40 - 3.40 min, held at 95% B for 0.45 min, then 95 - 5% B in 0.01 min, and the flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3 and mobile phase B was acetonitrile. The column used for chromatography was an Xbridge C18 2.1×50 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1500.)

[0159] General procedure for the preparation of A4BC1R3 (NUCC - 0226276) - ET37412 - 113

[0160]

Chemical Structure

[0161] 1 H NMR (ET37412-113-1, 400 MHz, methanol-d4) δ 0.99 (s, 9H), 1.95 - 2.27 (m, 2H), 2.45 (s, 3H), 2.55 - 2.82 (m, 2H), 3.39 (br s, 2H), 3.58 - 3.69 (m, 2H), 3.72 - 3.91 (m, 5H), 4.12 (br d, J = 4.16 Hz, 1H), 4.29 - 4.46 (m, 2H), 4.50 - 4.66 (m, 4H), 5.01 (br s, 1H), 6.58 (s, 1H), 6.80 - 6.94 (m, 3H), 7.13 - 7.19 (m, 2H), 7.23 (d, J = 8.56 Hz, 1H), 7.34 - 7.49 (m, 3H), 7.59 (q, J = 8.15 Hz, 2H), 7.74 (s, 1H), 8.85 (s, 1H) LCMS (ESI+): RT = 2.662 min, m / z 1043.3 (M+1) +

[0162] 5_95AB_6 min_MS1500-220-254-ELSD: LC / MS (Gradient: 5% B from 0.40 min, 5 - 95% B from 0.40 to 3.00 min, hold at 95% B for 1.00 min, then 95 - 5% B in 0.01 min, flow rate was 1.0 mL / min. Mobile phase A was 0.04% trifluoroacetic acid in water and mobile phase B was 0.02% trifluoroacetic acid in acetonitrile. The column used for chromatography was Luna C18 50×2.0 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1500.)

[0163] General procedure for the preparation of A4BC1R4 (NUCC - 0226302) - ET37412 - 131

[0164]

Chemical formula

[0165] 11H NMR (ET37412-131-y1, 400 MHz, chloroform-d) δ 1.94 - 1.99 (m, 1H), 2.40 - 2.55 (m, 1H), 2.60 - 2.72 (m, 2H), 3.49 - 3.58 (m, 2H), 3.61 - 3.69 (m, 2H), 3.76 - 3.80 (m, 5H), 4.02 - 4.12 (m, 2H), 4.54 (s, 2H), 4.77 (dd, J = 12.59, 5.38 Hz, 1H), 4.91 (s, 2H), 5.23 (br s, 1H), 6.49 (s, 1H), 6.69 (d, J = 8.68 Hz, 1H), 6.75 - 6.81 (m, 2H), 7.05 (dd, J = 12.41, 8.62 Hz, 3H), 7.14 (d, J = 8.56 Hz, 1H), 7.42 - 7.48 (m, 2H), 7.50 - 7.54 (m, 1H), 7.57 (br t, J = 5.26 Hz, 1H), 7.63 (dd, J = 8.31, 7.46 Hz, 1H), 7.70 (d, J = 1.83 Hz, 1H), 7.88 (s, 1H) LCMS (ESI+): RT = 3.346 min, m / z 944.1 (M+1) +

[0166] 5_95 CD_6 min_MS1500 - 220 - 254 - ELSD: LC / MS (Gradient was 5% B for 0.40 min, 5 - 95% B from 0.40 - 3.40 min, held at 95% B for 0.45 min, then 95 - 5% B in 0.01 min, and the flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3 and mobile phase B was acetonitrile. The column used for chromatography was an Xbridge C18 2.1×50 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1500.)

[0167] General procedure for the preparation of A4BC1R5 (NUCC - 0226277) - ET37412 - 112

[0168]

Chemical Structure

[0169] 1 H NMR (ET37412-112-1, 400 MHz, methanol-d4) δ 1.01 (s, 9H), 1.20 - 1.41 (m, 4H), 1.98 - 2.24 (m, 2H), 2.46 (s, 3H), 3.72 - 3.86 (m, 5H), 3.95 (br d, J = 12.35 Hz, 4H), 4.14 (br s, 2H), 4.25 (br s, 2H), 4.34 - 4.44 (m, 1H), 4.46 (br s, 1H), 4.56 - 4.66 (m, 2H), 4.73 (s, 1H), 5.01 (s, 2H), 6.58 (s, 1H), 6.85 (br d, J = 8.16 Hz, 3H), 6.96 - 7.09 (m, 2H), 7.15 (br d, J = 7.94 Hz, 2H), 7.23 (br d, J = 8.60 Hz, 1H), 7.45 (d, J = 7.72 Hz, 1H), 7.53 - 7.64 (m, 2H), 7.74 (s, 1H), 8.80 (s, 1H) LCMS (ESI+): RT = 3.067 min, m / z 1145.3 (M + 1) +

[0170] 5_95AB_6 min_MS1500-220-254-ELSD: LC / MS (Gradient: 5% B from 0.40 min, 5 - 95% B from 0.40 - 3.00 min, hold at 95% B for 1.00 min, then 95 - 5% B in 0.01 min; flow rate: 1.0 mL / min. Mobile phase A: 0.04% trifluoroacetic acid in water; mobile phase B: 0.02% trifluoroacetic acid in acetonitrile. Column used for chromatography: Luna C18 50×2.0 mm column (5 μm particles). Detection methods: diode array (DAD) and evaporative light scattering (ELSD) detection. MS mode: positive electrospray ionization. MS range: 100 - 1500.)

[0171] Synthesis of NUCC - 0226258, NUCC - 0226259, NUCC - 0226260, NUCC - 0226261

Chem.

Chem.

Chem.

[0172] Reactant LCMS: LCMS method: LCMS (ESI+): m / z = 231.1 (M+H)+, RT: 0.806 min

[0173] 5_95AB_2 min - 220 - 254 - ELSD: LC / MS (Gradient was 5% B in 0.40 min, 5 - 95% B from 0.40 - 3.00 min, held at 95% B for 1.00 min, then 95 - 5% B in 0.01 min, and the flow rate was 1.0 mL / min. Mobile phase A was 0.037% trifluoroacetic acid in water and mobile phase B was 0.018% trifluoroacetic acid in acetonitrile. The column used for chromatography was a Kinetex C18 50×2.1 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection and positive electrospray ionization. The MS range was 100 - 1000.)

[0174] General procedure for the preparation of compound 3 - ET32240 - 1059 [Chemical formula] A solution of compound 2 (180 g, 782.13 mmol, 1 equivalent), iodine (794.05 g, 3.13 mol, 630.20 mL, 4 equivalents), and pyridine (247.47 g, 3.13 mol, 252.52 mL, 4 equivalents) in chloroform (1 L) was stirred at 25 °C for 8 h. LCMS indicated that the reaction was complete. The mixture was poured into water (500 mL) and triturated with petroleum ether:ethyl acetate (10:1, 800 mL) to obtain compound 3 (210 g, 589.83 mmol, 75.41% yield) as a yellowish solid.

[0175] Reactant LCMS: LCMS method: LCMS (ESI+): m / z = 356.9 (M+H)+, RT: 0.890 min

[0176] 5_95AB_2 minutes - 220 - 254 - ELSD: LC / MS (Gradient: 5%B for 0.40 minutes, 5 - 95%B from 0.40 to 3.00 minutes, hold at 95%B for 1.00 minute, then 95 - 5%B in 0.01 minute, flow rate was 1.0 mL / min. Mobile phase A was 0.037% trifluoroacetic acid in water, and mobile phase B was 0.018% trifluoroacetic acid in acetonitrile. The column used for chromatography was a Kinetex C18 50×2.1 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection and positive electrospray ionization. The MS range was 100 - 1000.) 1 H NMR (400 MHz, methanol - d4) δ ppm 6.79 (s, 1H) 7.02 (d, J = 8.82 Hz, 1H) 7.95 - 7.99 (m, 1H)

[0177] General procedure for the preparation of compound 4 - ET32240 - 1078 [Chemical formula] To a solution of Cpd 3 (130 g, 365.13 mmol, 1 equivalent) in acetone (1 L), bromomethylbenzene (74.94 g, 438.16 mmol, 52.04 mL, 1.2 equivalents) and bromomethylbenzene (74.94 g, 438.16 mmol, 52.04 mL, 1.2 equivalents) were added potassium carbonate (100.93 g, 730.26 mmol, 2 equivalents). The mixture was stirred at 80 °C for 8 hours. TLC indicated that the reaction was complete. The mixture was poured into water (500 mL) and extracted with ethyl acetate (3 × 500 mL). The organic layer was dried over Na2SO4 and concentrated to obtain a crude product, which was purified by chromatography on silica, eluting with petroleum ether:ethyl acetate = 10:1 - 5:1, to give the desired product Cpd 4 (120 g, 268.96 mmol, yield 73.66%) as a yellowish solid. 11H NMR (400 MHz, chloroform-d) δ ppm 5.27 (s, 2H) 6.63 (s, 1H) 6.96 (d, J = 8.93 Hz, 1H) 7.10 - 7.39 (m, 8H) 7.40 - 7.46 (m, 2H) 8.08 (d, J = 8.93 Hz, 1H)

[0178] General procedure for the preparation of compound 5-ET32240-1083

Chemical formula

[0179] General procedure for the preparation of compound 6-ET32240-1131

Chemical formula

[0180] LCMS (ESI+): m / z = 527.2 (M+H)+, RT: 1.203 min

[0181] 5_95AB_2 min - 220 - 254 - ELSD: LC / MS (gradient was 5% B at 0.40 min, 5 - 95% B from 0.40 - 3.00 min, hold 95% B for 1.00 min, then 95 - 5% B in 0.01 min, flow rate was 1.0 mL / min. Mobile phase A was 0.037% trifluoroacetic acid in water and mobile phase B was 0.018% trifluoroacetic acid in acetonitrile. The column used for chromatography was a Kinetex C18 50×2.1 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection and positive electrospray ionization. The MS range was 100 - 1000.)

[0182] General procedure for the preparation of compound 7 - ET32240 - 1147

Chemical Structure

[0183] General procedure for the preparation of Compound Core A6 (NUCC-0226261)-ET32240-1158

Chemical Structure

[0184] Synthesis of Target D (NUCC - 0226219) and Target E (NUCC - 0226223)

Chemical Structure

Chemical Structure

Chemical Structure

[0185] Chemical Synthesis Experiment for the Largest - Scale Execution: General Procedure for the Preparation of Target D (NUCC - 0226219) - ET37412 - 4

Chemical Structure

[0186] 1 H NMR (ET37412-4-1, 400 MHz, DMSO-d6) δ 0.78 - 0.87 (m, 2H), 1.02 (br s, 4H), 1.36 (d, J = 12.28 Hz, 19H), 1.45 (s, 12H), 2.77 - 2.91 (m, 2H), 3.07 (br s, 2H), 3.24 (br s, 2H), 3.78 (s, 4H), 6.56 - 6.71 (m, 1H), 6.61 - 6.68 (m, 1H), 6.61 - 6.68 (m, 1H), 6.66 (s, 1H), 6.76 (br d, J = 8.23 Hz, 1H), 7.07 (br s, 1H), 7.64 (br s, 1H), 7.68 - 7.79 (m, 2H), 7.90 (s, 1H), 8.26 (br t, J = 5.60 Hz, 1H), 11.48 (br s, 1H) LCMS (ESI+): RT = 3.051 min, m / z 978.3 (M + 1) +

[0187] 5_95AB_6 min - 220 - 254 - ELSD LC / MS (Gradient: 5% B from 0.40 min, 5 - 95% B from 0.40 - 3.00 min, hold at 95% B for 1.00 min, then 95 - 5% B in 0.01 min, flow rate was 1.0 mL / min. Mobile phase A was 0.037% trifluoroacetic acid in water and mobile phase B was 0.018% trifluoroacetic acid in acetonitrile. The column used for chromatography was a Kinetex C18 50×2.1 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection and positive electrospray ionization. The MS range was 100 - 1000.)

[0188] General procedure for the preparation of Cpd 6 (NUCC - 0226260) - ET37412 -

[0189]

Chemical formula

[0190] 1H NMR (ET37412 - 34 - 1,400 MHz, chloroform - d) δ1.49(s,8H),1.56 - 1.65(m,4H),1.77 - 2.01(m,2H),3.27 - 3.53(m,5H),3.57 - 3.73(m,2H),3.96 - 4.22(m,3H)

[0191] General Procedure for the Preparation of Cpd 7-ET37412-71 [Chemical Formula] To a mixture of Core A6 (830 mg, 1.73 mmol, 1 equiv) and Cpd 6 (1.14 g, 2.60 mmol, purity 80%, 1.5 equiv) in DMF (5 mL), K2CO3 (359.40 mg, 2.60 mmol, 1.5 equiv) was added at once at 25 °C. The reaction mixture was stirred at 25 °C for 12 h. LCMS indicated the completion of the reaction. The reaction mixture was poured into water (10 mL) and extracted with ethyl acetate (20 mL). The organic layer was concentrated to give a residue, which was purified by silica gel chromatography eluting with petroleum ether / ethyl acetate = 1:0 to 0:1 to afford Cpd 7 (400 mg, yield 31%) as a colorless oil.

[0192] LCMS (ESI+): RT = 0.948 min, m / z 691.3 (M+1) +

[0193] 5 - 95 AB_2 min LC / MS (The column used for chromatography was Kinetex 5μm EVO C18 100A 2.1×30 mm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water and mobile phase B was 0.02% trifluoroacetic acid in HPLC grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B for 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B for 0.46 min, 95 - 5% B (1.61 - 1.50 min), and held at 5% B for 0.11 min. The flow rate was 1.5 mL / min.

[0194] General Procedure for the Preparation of Cpd 8-ET37412-48

[0195] [Chemical Formula] To a mixture of Cpd 7 (400 mg, 1 equiv) in dioxane (0.5 mL) was added HCl / dioxane (1 mL). The reaction mixture was stirred at 25 °C for 12 h. LCMS indicated that the reaction was complete. The reaction mixture was concentrated to give a residue, which was used in the next step without purification to afford Cpd 8 (330 mg, 91% yield, HCl) as a yellow oil.

[0196] LCMS (ESI+): RT = 0.773 min, m / z 647.7 (M+1) +

[0197] 5 - 95 AB_2 min: LC / MS (The column used for chromatography was Kinetex 5μm EVO C18 100A 2.1×30 mm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water and mobile phase B was 0.02% trifluoroacetic acid in HPLC grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B for 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B for 0.46 min, 95 - 5% B (1.61 - 1.50 min), and held at 5% B for 0.11 min. The flow rate was 1.5 mL / min.

[0198] General procedure for the preparation of target E (NUCC - 0226223) - ET37412 - 52

[0199]

Chemical Structure

[0200] 1 H NMR (ET37412-52-P1B, 400 MHz, methanol-d4) δ 1.17 - 1.28 (m, 2H), 1.31 - 1.39 (m, 2H), 1.45 (d, J = 5.95 Hz, 6H), 1.61 - 1.71 (m, 2H), 2.97 (br s, 2H), 3.02 - 3.12 (m, 1H), 3.18 - 3.27 (m, 1H), 3.41 - 3.53 (m, 1H), 3.56 - 3.68 (m, 1H), 3.79 (s, 3H), 3.88 (s, 2H), 3.91 (s, 3H), 3.96 - 4.02 (m, 2H), 4.91 (dd, J = 12.24, 6.06 Hz, 1H), 5.98 (d, J = 11.03 Hz, 1H), 6.24 (d, J = 11.03 Hz, 1H), 6.58 (s, 1H), 6.74 - 6.84 (m, 3H), 6.99 (d, J = 8.38 Hz, 2H), 7.17 (dd, J = 16.21, 8.49 Hz, 4H), 7.26 (dd, J = 17.42, 8.60 Hz, 3H), 7.56 - 7.61 (m, 1H), 7.63 - 7.70 (m, 2H), 7.75 (d, J = 1.76 Hz, 1H) LCMS (ESI+): RT = 2.966 min, m / z 1085.0 (M+1) +

[0201] 5_95AB_6 min_MS1500-220-254-ELSD: LC / MS (Gradient: 5% B from 0.40 min, 5 - 95% B from 0.40 to 3.00 min, hold at 95% B for 1.00 min, then 95 - 5% B in 0.01 min, flow rate was 1.0 mL / min. Mobile phase A was 0.04% trifluoroacetic acid in water and mobile phase B was 0.02% trifluoroacetic acid in acetonitrile. The column used for chromatography was Luna C18 50×2.0 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1500.)

[0202] Synthesis of R5 (NUCC-0226278)

Chemical Structure

Chemical Structure

Chemical Structure

[0203] General procedure for the preparation of 2-(aminomethyl)-5-(4-methylthiazol-5-yl)phenol (D3) - ET39710-106

Chemical formula

[0204] (2S,4R)-tert-Butyl 4-hydroxy-2-((2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidine-1-carboxylate (D4) - General procedure for the preparation of ET39710-107

Chem.

[0205] General procedure for the preparation of (2S,4R)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide hydrochloride (D5)-ET39710-112

Chemical formula

[0206] General procedure for the preparation of tert-butyl ((S)-1-((2S,4R)-4-hydroxy-2-((2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)carbamate (D6) - ET39710-113 To a solution of (2S,4R)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide hydrochloride (D5) (16 g, 43.26 mmol) in DMF (160 mL) were added (2S)-2-(tert-butoxycarbonylamino)-3,3-dimethyl-butanoic acid (10.01 g, 43.26 mmol), DIEA (116.77 g, 129.78 mmol) and HATU (19.74 g, 51.91 mmol) at 0 °C. The mixture was stirred at 20 °C for 1 h. LCMS indicated that the reaction was complete. The mixture was poured into ice water (100 mL) and stirred for 5 min. The mixture was extracted with EA (3 × 150 mL).

[0207] The combined organic layers were washed with brine (300 mL), dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (eluting with PE / EA = 50:1 to 0:1) to give tert-butyl ((S)-1-((2S,4R)-4-hydroxy-2-((2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)carbamate (D6) (12.1 g, yield 51.17%) as a yellow solid. 1 H NMR (400 MHz, MeOD-d4) δ = 8.85 (s, 1H), 8.74 (dd, J = 1.4, 4.3 Hz, 1H), 8.43 (dd, J = 1.2, 8.3 Hz, 1H), 7.52 (dd, J = 4.8, 8.6 Hz, 1H), 7.36 (d, J = 8.1 Hz, 1H), 6.92 - 6.87 (m, 2H), 4.60 (s, 1H), 4.50 (br s, 1H), 4.45 - 4.32 (m, 2H), 4.29 (s, 1H), 3.91 - 3.76 (m, 2H), 2.49 - 2.47 (m, 3H), 2.25 - 2.06 (m, 2H), 1.48 - 1.42 (m, 9H), 1.00 (s, 9H)

[0208] (2S,4R)-1-[(2S)-2-Amino-3,3-dimethyl-butanoyl]-4-hydroxy-N-[[2-hydroxy-4-(4-methylthiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide (D7) - General procedure for the preparation of ET39710-115 [Chemical formula] A mixture of tert-butyl ((S)-1-((2S,4R)-4-hydroxy-2-((2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)carbamate (D6) (12 g, 21.95 mmol) in HCl / dioxane (120 mL, 4 M) was stirred at 20 °C for 20 minutes. LCMS indicated that the reaction was complete. The mixture was concentrated under reduced pressure to give (2S,4R)-1-[(2S)-2-amino-3,3-dimethyl-butanoyl]-4-hydroxy-N-[[2-hydroxy-4-(4-methylthiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide (D7) (9.95 g, yield 93.84%, HCl salt) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ = 9.05 (s, 1H), 8.63 (t, J = 6.1 Hz, 1H), 8.15 (br d, J = 3.9 Hz, 3H), 7.32 (d, J = 7.8 Hz, 1H), 6.98 (d, J = 2.0 Hz, 1H), 6.81 (dd, J = 2.0, 7.8 Hz, 1H), 4.57 (t, J = 8.3 Hz, 1H), 4.37 (br s, 1H), 4.33 - 4.23 (m, 1H), 4.20 - 4.10 (m, 1H), 3.90 (br d, J = 4.9 Hz, 1H), 3.77 (br d, J = 10.8 Hz, 1H), 3.64 - 3.51 (m, 1H), 2.12 (br dd, J = 7.8, 12.7 Hz, 1H), 1.91 (s, 1H), 1.59 (s, 4H), 1.01 (s, 9H)

[0209] (2S,4R)-1-((S)-2-(1-Fluorocyclopropanecarboxamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(2-hydroxy-4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (NUCC-0226278) - General procedure for the preparation of ET39710-118

Chemical Structure

[0210] LCMS (ESI+): m / z = 533.0 (M+H) + , RT: 0.832 min 11H NMR (400 MHz, DMSO-d6) δ = 9.97 - 9.71 (m, 1H), 9.09 - 8.94 (m, 1H), 8.66 - 8.45 (m, 1H), 7.36 - 7.23 (m, 2H), 7.01 - 6.87 (m, 1H), 6.87 - 6.80 (m, 1H), 4.70 - 4.43 (m, 6H), 4.32 - 4.03 (m, 3H), 3.69 - 3.56 (m, 2H), 2.13 - 2.03 (m, 1H), 1.96 - 1.87 (m, 1H), 1.44 - 1.30 (m, 2H), 1.27 - 1.17 (m, 2H), 1.01 - 0.89 (m, 9H) Prep-HPLC method: Equipment: Shimadzu LC-8A preparative HPLC Column: Phenomenex luna C18 250×50 mm×10 μm Mobile phase: A = H2O (0.04% HCl) and B = CH3CN Gradient: 25% - 45% B in 20 minutes Flow rate: 80 mL / min Wavelength: 220 and 254 nm SFC method: Column: Chiralpak AD-3, 50×4.6 mm I.D., 3 μm Mobile phase: A = CO2 B = EtOH (0.1% IPAm, v / v) Gradient:

Table 1

[0211] Synthesis of A04B01C01D01 (NUCC-0226545)

Chem.

Chem.

[0212] Method 2: 50_100 CD_6 minutes - 220 - 254 - ELSD For LC / MS (gradient was 50% B in 0.40 minutes, 50 to 100% B from 0.40 to 3.40 minutes, held at 100% B for 0.45 minutes, then 100 to 50% B in 0.01 minute, and the flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3, and mobile phase B was acetonitrile. The column used in chromatography was an Xbridge Shield RP18 2.1×50 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection as well as positive electrospray ionization. The MS range was 100 to 1000.

[0213] Experiment for the largest-scale execution: General procedure for the preparation of compound 2-ET42365-46

[0214] [Chemical formula] To a solution of AlCl3 (5.12 g, 38.42 mmol, 2.10 mL, 1.2 equiv) in DCM (50 mL), acetyl chloride (3.77 g, 48.03 mmol, 3.43 mL, 1.5 equiv) was added dropwise at 5 - 10 °C. The reaction mixture was stirred at 5 °C for 15 minutes, then compound 1 (5 g, 32.02 mmol, 1 equiv) was added dropwise at 0 - 10 °C, and the reaction mixture was stirred at 20 °C for 6 hours. TLC (petroleum ether / ethyl acetate = 3 / 1) indicated that the starting material was consumed and two new peaks were formed. The reaction mixture was poured into 200 mL of ice water. The aqueous layer was extracted with dichloromethane (2 × 100 mL). The combined organic phases were washed with water (100 mL), dried over sodium sulfate, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography on silica gel (eluting with petroleum ether / ethyl acetate = 50 / 1 - 10 / 1) to give compound 2 (4.5 g, yield 67.37%) as a colorless oil.

[0215] 1 H NMR (ET42365 - 46 - P1A, 400 MHz, chloroform - d) δ 2.52 (d, J = 1.63 Hz, 3H), 3.82 (s, 3H), 3.84 (s, 3H), 6.22 - 6.28 (m, 2H) 19 F NMR (ET42365 - 46 - P1A, 400 MHz, CHLOROFORM - d) δ - 112.015

[0216] General procedure for the preparation of compound 3 - ET42365 - 68

[0217]

Chemical Structure

[0218] 1 H NMR (ET42365 - 68 - P1H, 400 MHz, DMSO - d6) δ 2.53 (d, J = 6.80 Hz, 3H), 6.11 (d, J = 2.19 Hz, 1H), 6.19 (dd, J = 14.03, 2.19 Hz, 1H), 11.06 (s, 1H), 13.05 (s, 1H)

[0219] General procedure for the preparation of compound 4 - ET43588 - 7

Chemical formula

[0220] LCMS (ESI+): RT: 0.773 min, m / z = 249.0 (M+H) + 。

[0221] 5 - 95 AB_2 min: LC / MS (The column used for chromatography was Chromolith® RP - 18e 25 - 2 mm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water and mobile phase B was 0.02% trifluoroacetic acid in HPLC - grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B for 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95 - 100% B (0.70 - 1.15 min), 5% B at 1.16 min and held at 5% B for 0.34 min. The flow rate was 1.5 mL / min.

[0222] General procedure for the preparation of compound 6 - ET43588 - 20

[0223]

Chem.

[0224] 1 H NMR (ET43588 - 20 - P1H, 400 MHz, MeOD) δ 6.72 - 6.80 (m, 2H)

[0225] General procedure for the preparation of compound 7 - ET43588 - 21

Chemical formula

[0226] 1HNMR(ET43588 - 20 - P1H, 400 MHz, CDCl3) δ 5.25 (s, 2H) 6.66 (s, 1H) 6.77 (d, J = 12.17 Hz, 1H) 7.39 - 7.48 (m, 4H)

[0227] General procedure for the preparation of compound 8 - ET43588 - 38

[0228] The reactions were carried out in parallel and combined for purification.

[0229]

Chemical formula

[0230] 1 H NMR(ET43588 - 38 - PH1, 400 MHz, CDCl3) δ ppm 5.15 (s, 2H) 6.63 (s, 1H) 6.89 (d, J = 12.13 Hz, 1H) 7.19 (d, J = 8.38 Hz, 2H) 7.35 (d, J = 8.50 Hz, 2H) 7.51 (dd, J = 8.38, 1.88 Hz, 1H) 7.61 (d, J = 8.25 Hz, 1H) 7.77 (d, J = 1.88 Hz, 1H)

[0231] General procedure for the preparation of A04B01C01D01-ET43588-57

[0232] [Chemical formula] To a suspension of Compound 8 (0.16 g, 290.26 μmol) in ethanol (1.6 mL), CH3NHNH2 (200.59 mg, 1.74 mmol, 229.24 μL, purity 40%) was added at 25 °C. Then the reaction mixture was stirred and heated at 80 °C for 4 hours. LCMS indicated that the reaction was complete. The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC to obtain A04B01C01D01 (17.7 mg, yield 10.5%) as a white solid.

[0233] 1 H NMR (ET43588-57-P1H1, 400 MHz, CDCl3) δ 3.85 (s, 3H) 5.04 (s, 2H) 5.27 (s, 1H) 6.53 (d, J = 11.21 Hz, 1H) 6.65 (s, 1H) 7.17 (d, J = 8.23 Hz, 2H) 7.34 (d, J = 8.34 Hz, 2H) 7.48 - 7.55 (m, 1H) 7.57 - 7.64 (m, 1H) 7.77 (s, 1H) Preparative HPLC method: Equipment: Gilson 281 semi-preparative HPLC system Mobile phase: A: 10 mM NH4HCO3 in H2O; B: CAN Column: Waters Xbridge BEH C18 100×25 mm×5 μm Flow rate: 25 mL / min Monitoring wavelength: 220 and 254 nm [Table 2] LCMS (ESI+): RT: 2.919 min, m / z = 579.1 (M + H) +

[0234] 50_100 CD_6 min - 220 - 254 - ELSD LC / MS (Gradient: 50% B in 0.40 min, 50 - 100% B from 0.40 to 3.40 min, hold at 100% B for 0.45 min, then 100 - 50% B in 0.01 min; flow rate: 0.8 mL / min. Mobile phase A: H2O + 10 mM NH4HCO3; mobile phase B: acetonitrile. Column used for chromatography: Xbridge Shield RP18 2.1×50 mm column (5 μm particles). Detection methods: diode array (DAD) and evaporative light scattering (ELSD) detection and positive electrospray ionization. MS range: 100 - 1000.)

[0235] Synthesis of NUCC - 0226530 (ET42365 - 98 - 1), NUCC - 0226528 (ET42365 - 86 - 1) and NUCC - 0226520 (ET42365 - 63 - 1)

Chemical Structure

Chemical Structure

Chemical Structure

[0236] 50_100 CD_6 min - 220 - 254 - ELSD: LC / MS (Gradient: 50% B at 0.40 min, 50 - 100% B from 0.40 to 3.40 min, hold at 100% B for 0.45 min, then 100 - 50% B in 0.01 min, flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3, mobile phase B was acetonitrile. The column used for chromatography was an Xbridge C18 2.1×50 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0237] 5_95 CD_6 min - 220 - 254 - ELSD: LC / MS (Gradient: 5% B at 0.40 min, 5 - 95% B from 0.40 to 3.40 min, hold at 95% B for 0.45 min, then 95 - 5% B in 0.01 min, flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3, mobile phase B was acetonitrile. The column used for chromatography was an Xbridge C18 2.1×50 mm column (5 μm particles). The detection method was diode array (DAD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0238] Experiment for the largest - scale execution: General procedure for the preparation of compound 2 - ET42365 - 29

[0239] [Chemical formula] A solution of compound 1 (10 g, 43.45 mmol, 1 equiv) and pyridine (13.75 g, 173.81 mmol, 14.03 mL, 4 equiv) in DCM (150 mL) was added dropwise with Tf2O (18.39 g, 65.18 mmol, 10.75 mL, 1.5 equiv) at 0 °C, and the reaction mixture was stirred at 20 °C for 12 h. LCMS indicated that the starting material was consumed and a new peak with the desired product Ms was detected. The reaction mixture was diluted with water (300 mL) and extracted with DCM (3 × 100 mL). The organic layers were separated, combined organic layers were washed with brine (150 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography on silica gel (eluting with petroleum ether / ethyl acetate = 50 / 1 to 10 / 1) to give compound 2 (12 g, yield 72.43%) as a white solid.

[0240] 1 H NMR (ET42365 - 29 - P1A, 400 MHz, chloroform - d) δ 6.79 (s, 1H), 7.42 (dd, J = 8.88, 2.25 Hz, 1H), 7.56 (d, J = 2.25 Hz, 1H), 8.34 (d, J = 8.88 Hz, 1H)

[0241] General procedure for the preparation of compound 3 - ET42365 - 43

Chemical Structure

[0242] LCMS (ESI+): RT = 0.827 min, m / z 396.2 (M + H) +

[0243] 5 - 95 AB_2 min: LC / MS (The column used for chromatography was an Agilent Poroshell SB - C18 3.0×30 mm, 2.7 μm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water and mobile phase B was 0.02% trifluoroacetic acid in HPLC - grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B (0.70 - 1.16 min), 95 - 5% B (1.16 - 1.50 min). The flow rate was 1.5 mL / min.

[0244] General procedure for the preparation of Compound 4 - ET42365 - 49

[0245]

Chemical Structure

[0246] 1 H NMR (ET42365 - 49 - P1A, 400 MHz, chloroform - d) δ 5.70 (br d, J = 5.04 Hz, 1H), 5.77 (d, J = 5.48 Hz, 1H), 6.60 - 6.66 (m, 2H), 7.29 - 7.43 (m, 10H), 7.88 (d, J = 8.99 Hz, 1H)

[0247] General procedure for the preparation of compound 5 - ET42365 - 65

Chemical formula

[0248] LCMS (ESI+): RT = 0.943 min, m / z 574.3 (M+H) +

[0249] 5 - 95 AB_2 min: LC / MS (The column used for chromatography was an Agilent Poroshell SB-C18 3.0×30 mm, 2.7 μm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water and mobile phase B was 0.02% trifluoroacetic acid in HPLC grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B (0.70 - 1.16 min), 95 - 5% B (1.16 - 1.50 min). The flow rate was 1.5 mL / min.

[0250] General procedure for the preparation of compound 6-ET42365-70

[0251]

Chemical Structure

[0252] LCMS (ESI+): RT = 0.943 min, m / z 602.3 (M+H) +

[0253] 5 - 95 AB_2 min: LC / MS (The column used for chromatography was Agilent Poroshell SB-C18 3.0×30 mm, 2.7 μm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in HPLC grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B (0.70 - 1.16 min), 95 - 5% B (1.16 - 1.50 min). The flow rate was 1.5 mL / min.

[0254] General procedure for the preparation of A01B01C04D01-ET42365-75

[0255]

Chemical Structure

[0256] LCMS (ESI+): RT = 0.800 min, m / z 436.2 (M+H) +

[0257] 5 - 95 AB_2 min: LC / MS (The column used for chromatography was an Agilent Poroshell SB-C18 3.0×30 mm, 2.7 μm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water and mobile phase B was 0.02% trifluoroacetic acid in HPLC grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B (0.70 - 1.16 min), 95 - 5% B (1.16 - 1.50 min). The flow rate was 1.5 mL / min.

[0258] General procedure for the preparation of A02B01C04D01-ET42365-88

[0259]

Chemical Structure

[0260] LCMS (ESI+): RT = 0.833 min, m / z 470.1 (M+H) +

[0261] 5 - 95 AB_2 min: LC / MS (The column used for chromatography was Agilent Poroshell SB-C18 3.0×30 mm, 2.7 μm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water and mobile phase B was 0.02% trifluoroacetic acid in HPLC grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B for 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B (0.70 - 1.16 min), 95 - 5% B (1.16 - 1.50 min). The flow rate was 1.5 mL / min.

[0262] General procedure for the preparation of A02B01C09D01 - ET42365 - 98

[0263]

Chemical Structure

[0264] 1 H NMR (ET42365-98-P1, 400 MHz, chloroform-d) δ 2.53 (s, 6H), 3.79 (s, 3H), 4.86 (s, 1H), 6.50 (s, 1H), 7.20 (s, 1H), 7.37 (dd, J = 8.19, 1.94 Hz, 1H), 7.55 - 7.61 (m, 2H) LCMS (ESI+): RT = 2.940 min, m / z 498.1 (M + H) +

[0265] 50_100CD_6 min - 220 - 254 - ELSD: LC / MS (gradient was 50% B at 0.40 min, 50 - 100% B from 0.40 - 3.40 min, hold 100% B for 0.45 min, then 100 - 50% B in 0.01 min, flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3 and mobile phase B was acetonitrile. The column used for chromatography was an Xbridge C18 2.1×50 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1000.

[0266] General procedure for the preparation of A02B01C04D01-ET42365-86

[0267]

Chem.

[0268] 1 H NMR (ET42365-86-P1B, 400 MHz, chloroform-d) δ 3.86 (s, 3H), 4.17 (br s, 2H), 4.82 (s, 1H), 6.56 (s, 1H), 7.20 (s, 1H), 7.53 - 7.57 (m, 1H), 7.73 (d, J = 8.28 Hz, 1H), 7.76 (d, J = 1.63 Hz, 1H) LCMS (ESI+): RT = 3.527 min, m / z 470.1 (M + H) +

[0269] 5_95CD_6 min - 220 - 254 - ELSD: LC / MS (gradient was 5% B at 0.40 min, 5 - 95% B from 0.40 - 3.40 min, hold at 95% B for 0.45 min, then 95 - 5% B in 0.01 min, flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3 and mobile phase B was acetonitrile. The column used for chromatography was an Xbridge C18 2.1×50 mm column (5 μm particles). The detection method was diode array (DAD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0270] General procedure for the preparation of A01B01C04D01-ET42365-63

[0271] [Chemistry] To a solution of compound 6 (100 mg, 166.12 μmol, 1 equiv) in MeOH (1 mL), concentrated aqueous HCl (504.75 mg, 4.98 mmol, purity 36%, 30 equiv) was added, and the reaction mixture was stirred at 80 °C for 8 h. LCMS indicated that the starting material was consumed and a new peak with the desired product Ms was detected. The reaction mixture was concentrated to obtain the crude product. The crude product was purified by Prep-HPLC (column: Phenomenex Gemini-NX C18 75×30 mm×3 μm; mobile phase: [water (10 mM NH4HCO3)-ACN]; B%: 45% - 65%, 8 min) to give A01B01C04D01 (21.5 mg, yield 29.61%) as a peach-colored solid.

[0272] 1 H NMR (ET42365-63-P1B, 400 MHz, chloroform-d) δ 3.76 (br s, 2H), 3.85 (s, 3H), 4.87 (s, 1H), 6.48 (d, J = 8.25 Hz, 1H), 6.55 (s, 1H), 7.05 (d, J = 8.25 Hz, 1H), 7.56 (dd, J = 8.19, 1.81 Hz, 1H), 7.69 (d, J = 8.25 Hz, 1H), 7.78 (d, J = 1.63 Hz, 1H) LCMS (ESI+): RT = 3.311 min, m / z 436.1 (M+H) +

[0273] 5_95 CD_6 min - 220 - 254 - ELSD: LC / MS (gradient was 5% B at 0.40 min, 5 - 95% B from 0.40 - 3.40 min, hold at 95% B for 0.45 min, then 95 - 5% B in 0.01 min, flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3 and mobile phase B was acetonitrile. The column used for chromatography was an Xbridge C18 2.1×50 mm column (5 μm particles). The detection method was diode array (DAD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0274] Synthesis of NUCC-0226529 (ET42365-96-1) and NUCC-0226527 (ET42365-83-1)

Chem.

Chem.

[0275] 50_100 CD_6 min - 220 - 254 - ELSD: LC / MS (The gradient was 50% B in 0.40 min, 50 - 100% B from 0.40 - 3.40 min, hold at 100% B for 0.45 min, then 100 - 50% B in 0.01 min, and the flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3, and mobile phase B was acetonitrile. The column used for chromatography was an Xbridge C18 2.1×50 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0276] 5_95CD_6 min - 220 - 254 - ELSD: LC / MS (Gradient was 5% B from 0.40 min, 5 - 95% B from 0.40 - 3.40 min, held at 95% B for 0.45 min, then 95 - 5% B in 0.01 min, and the flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3 and mobile phase B was acetonitrile. The column used for chromatography was an Xbridge C18 2.1×50 mm column (5 μm particles). The detection method was diode array (DAD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0277] Experiment for the largest scale execution: General procedure for the preparation of compound 7 - ET42365 - 79

[0278] [Chemical formula] Paraformaldehyde (249.40 mg, 8.31 mmol, 10 equiv) was added to a solution of compound 6 (500 mg, 830.61 μmol, 1 equiv) in AcOH (10 mL). After stirring at 20 °C for 1.5 h, NaBH3 (260.99 mg, 4.15 mmol, 5 equiv) was added. The mixture was stirred at 25 °C for 12 h. LCMS indicated that the starting material was consumed and a new peak with the desired product Ms was detected. The reaction mixture was concentrated to obtain the crude product, which was diluted with water (50 mL) and extracted with ethyl acetate (3×50 mL). The organic layers were separated, combined organic layers were washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain crude compound 7 (750 mg, yield 58.63%) as a yellow solid. The crude product was used directly in the next step without further purification.

[0279] LCMS (ESI+): RT = 0.969 min, m / z 616.3 (M + H) +

[0280] 5 - 95 AB_2 min: For LC / MS, the column used in chromatography was Agilent Poroshell SB - C18 3.0×30 mm, 2.7 μm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in HPLC - grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B (0.70 - 1.16 min), 95 - 5% B (1.16 - 1.50 min). The flow rate was 1.5 mL / min.

[0281] General procedure for the preparation of A01B01C03D01 - ET42365 - 89

[0282]

Chemical formula

[0283] LCMS (ESI+): RT = 0.852 min, m / z 450.2 (M + H) +

[0284] 5 - 95 AB_2 min: The column used in LC / MS was an Agilent Poroshell SB - C18 3.0×30 mm, 2.7 μm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in HPLC - grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B (0.70 - 1.16 min), 95 - 5% B (1.16 - 1.50 min). The flow rate was 1.5 mL / min.

[0285] General procedure for the preparation of A02B01C05D01 - ET42365 - 96

[0286]

Chemical formula

[0287] 1 H NMR (ET42365 - 96 - P1A, 400 MHz, chloroform - d) δ 2.39 (s, 3H), 3.86 (s, 3H), 4.89 (s, 1H), 6.55 (s, 1H), 7.22 (s, 1H), 7.55 - 7.60 (m, 1H), 7.64 - 7.69 (m, 1H), 7.78 (d, J = 1.88 Hz, 1H) LCMS (ESI+): RT = 2.520 min, m / z 484.1 (M+H) + 。

[0288] 50_100 CD_6 min - 220 - 254 - ELSD: LC / MS (Gradient is 50% B in 0.40 min, 50 - 100% B from 0.40 to 3.40 min, hold 100% B for 0.45 min, then 100 - 50% B in 0.01 min, and the flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3 and mobile phase B was acetonitrile. The column used for chromatography was an Xbridge C18 2.1×50 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0289] General procedure for the preparation of A01B01C03D01 - ET42365 - 83

[0290]

Chem.

[0291] 11H NMR (ET42365-83-P1A, 400 MHz, chloroform-d) δ 2.84 (s, 3H), 3.85 (s, 3H), 4.78 (s, 1H), 6.42 (br d, J = 8.63 Hz, 1H), 6.55 (s, 1H), 7.16 (d, J = 8.38 Hz, 1H), 7.52 (br d, J = 7.88 Hz, 1H), 7.66 - 7.76 (m, 2H) LCMS (ESI+): RT = 3.551 min, m / z 450.1 (M + H) +

[0292] 5_95 CD_6 min - 220 - 254 - ELSD: LC / MS (The gradient was 5% B for 0.40 min, 5 - 95% B from 0.40 to 3.40 min, hold 95% B for 0.45 min, then 95 - 5% B in 0.01 min, and the flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3 and mobile phase B was acetonitrile. The column used for chromatography was an Xbridge C18 2.1×50 mm column (5 μm particles). The detection method was diode array (DAD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0293] Synthesis of NUCC - 0226501 (ET42365 - 34 - 1)

Chemical Structure

Chemical Structure

[0294] 50_100 CD_6 min - 220 - 254 - ELSD: LC / MS (The gradient was 50% B in 0.40 min, 50 - 100% B from 0.40 - 3.40 min, hold 100% B for 0.45 min, then 100 - 50% B in 0.01 min, and the flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3, and mobile phase B was acetonitrile. The column used in chromatography was Xbridge C18 2.1×50 mm column (5 - μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0295] Experiment for the largest - scale execution: General procedure for the preparation of Core A3_2 - ET43365 - 14

[0296]

Chemical formula

[0297] LCMS (ESI+): RT = 0.862 min, m / z 493.2 (M+H) +

[0298] 5 - 95 AB_2 min: LC / MS (The column used for chromatography was Agilent Poroshell SB-C18 3.0 × 30 mm, 2.7 μm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in HPLC grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B (0.70 - 1.16 min), 95 - 5% B (1.16 - 1.50 min). The flow rate was 1.5 mL / min.

[0299] General procedure for the preparation of core A3_5-ET42365-22

[0300]

Chemical Structure

[0301] 1 H NMR (ET42365-22-P1A, 400 MHz, chloroform-d) δ 1.78 (s, 3H), 3.78 (s, 3H), 3.87 (s, 3H), 7.06 (d, J = 8.63 Hz, 1H), 7.37 (d, J = 8.63 Hz, 1H), 7.46 (dd, J = 8.25, 1.75 Hz, 1H), 7.56 (d, J = 8.25 Hz, 1H), 7.67 (d, J = 1.63 Hz, 1H)

[0302] General procedure for the preparation of A02B01C02D01_isomer-ET42365-34

Chemical Structure

[0303] 1 H NMR (ET42365-34-P1A, 400 MHz, chloroform-d) δ 3.82 (d, J = 4.75 Hz, 6H), 4.97 (s, 1H), 6.76 (d, J = 8.76 Hz, 1H), 7.29 (d, J = 8.63 Hz, 1H), 7.53 (dd, J = 8.13, 2.00 Hz, 1H), 7.66 (d, J = 8.25 Hz, 1H), 7.75 (d, J = 1.88 Hz, 1H) LCMS (ESI+): RT = 2.439 min, m / z 485.1 (M+H) +

[0304] 50_100CD_6 min - 220 - 254 - ELSD:LC / MS (Gradient: 50% B at 0.40 min, 50 - 100% B from 0.40 to 3.40 min, hold at 100% B for 0.45 min, then 100 - 50% B in 0.01 min, flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3, and mobile phase B was acetonitrile. The column used for chromatography was an Xbridge C18 2.1×50 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0305] Synthesis of NUCC - 0226500 (ET42365 - 41 - 1) [Chemical formula] Synthesis scheme: Preparation of A02B01C02D01 (NUCC - 0226500) [Chemical formula] Chemical synthesis 5 - 95AB_2 min:LC / MS (The column used for chromatography was an Agilent Poroshell SB - C18 3.0×30 mm, 2.7 μm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in HPLC - grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B (0.70 - 1.16 min), 95 - 5% B (1.16 - 1.50 min). The flow rate was 1.5 mL / min.)

[0306] 50_100CD_6 min - 220 - 254 - ELSD: LC / MS (Gradient: 50% B at 0.40 min, 50 - 100% B from 0.40 - 3.40 min, hold 100% B for 0.45 min, then 100 - 50% B in 0.01 min, flow rate 0.8 mL / min. Mobile phase A is H2O + 10 mM NH4HCO3, mobile phase B is acetonitrile. The column used for chromatography was an Xbridge C18 2.1×50 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0307] Experiment for the largest - scale execution: General procedure for the preparation of Core A3_4 - ET43365 - 31

[0308] [Chemical formula] To a solution of Core A3_1 (700 mg, 1.46 mmol, 1 equiv) and N - isopropylpropan - 2 - amine (295.89 mg, 2.92 mmol, 413.26 μL, 2 equiv) in CHCl3 (25 mL) was added NCS (214.76 mg, 1.61 mmol, 1.1 equiv) at 0 °C, and the reaction mixture was stirred at 25 °C for 24 h. LCMS indicated that approximately 28% of the starting material remained and 64% of the desired product was detected. The reaction mixture was concentrated under high vacuum to obtain the crude product. The crude product was purified by column chromatography on silica gel (eluting with petroleum ether / ethyl acetate = 50 / 1 - 10 / 1) to give Core A3_4 (200 mg, yield 25.32%) as a white solid.

[0309] 11H NMR (ET42365-31-P1A, 400 MHz, chloroform-d) δ 1.78 (s, 3H), 3.83 (s, 3H), 5.92 (s, 1H), 6.51 (s, 1H), 7.42 (s, 1H), 7.50 (dd, J = 8.25, 1.50 Hz, 1H), 7.61 (d, J = 8.38 Hz, 1H), 7.72 (d, J = 1.38 Hz, 1H)

[0310] General procedure for the preparation of Core A3_3-ET42365-40

Chemical Structure

[0311] LCMS (ESI+): RT = 0.892 min, m / z 527.2 (M + H) +

[0312] 5 - 95 AB_2 min: The column used in LC / MS was an Agilent Poroshell SB - C18 3.0×30 mm, 2.7 μm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in HPLC - grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B (0.70 - 1.16 min), 95 - 5% B (1.16 - 1.50 min). The flow rate was 1.5 mL / min.

[0313] General procedure for the preparation of A02B01C02D01 - ET42365 - 41

[0314]

Chemical formula

[0315] 11H NMR (ET42365-41-P1A, 400 MHz, chloroform-d) δ 3.59 (s, 3H), 3.87 (s, 3H), 5.06 (s, 1H), 6.61 (s, 1H), 7.34 (s, 1H), 7.56 - 7.62 (m, 1H), 7.65 - 7.71 (m, 1H), 7.80 (s, 1H) LCMS (ESI+): RT = 2.501 min, m / z 485.1 (M + H) +

[0316] 50_100 CD_6 min - 220 - 254 - ELSD: LC / MS (Gradient was 50% B at 0.40 min, 50 - 100% B from 0.40 to 3.40 min, held at 100% B for 0.45 min, then 100 - 50% B in 0.01 min, and the flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3 and mobile phase B was acetonitrile. The column used for chromatography was an Xbridge C18 2.1×50 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0317] Synthesis of Series 10 - D, E, F, M, K, L

Chemical Structure

[0318]

Chemical Structure

Chemical Structure

[0319] 1 H NMR: ET37412-154-HNMRP1 (400 MHz, CDCl3) δ 0.90 (d, J = 6.6 Hz, 6H), 1.50 (br d, J = 6.3 Hz, 3H), 1.85 (quind, J = 6.7, 13.4 Hz, 1H), 2.46 (d, J = 7.2 Hz, 2H), 5.08 (s, 1H), 3.81 (s, 3H), 5.32 (q, J = 6.3 Hz, 1H), 6.53 (s, 1H), 6.60 (d, J = 8.7 Hz, 1H), 7.15 - 7.05 (m, 5H), 7.60 - 7.53 (m, 1H), 7.67 - 7.62 (m, 1H), 7.83 (s, 1H) LCMS method: LCMS (ESI+): m / z = 597.3 (M + H) + , RT: 3.534 minutes

[0320] 5_95AB_6 min - 220 - 254 - ELSD: LC / MS (Gradient was 5% B from 0.40 min, 5 - 95% B from 0.40 - 3.00 min, held at 95% B for 1.00 min, then 95 - 5% B in 0.01 min, and the flow rate was 1.0 mL / min. Mobile phase A was 0.037% trifluoroacetic acid in water, and mobile phase B was 0.018% trifluoroacetic acid in acetonitrile. The column used for chromatography was a Kinetex C18 50×2.1 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection and positive electrospray ionization. The MS range was 100 - 1000.)

[0321] General procedure for the preparation of Cpd 2 - NotebookPage: ET37412 - 181

[0322] [Chemical formula] To a solution of Cpd 1 (1 g, 5.67 mmol) in tetrahydrofuran (10 mL), borane - tetrahydrofuran complex (6.81 mL, 6.81 mmol) was added at - 40 °C. To the mixture reactant, chloro - bis[(1R,2S,3R,5R) - 2,6,6 - trimethylnorpinan - 3 - yl]borane (2.18 g, 6.81 mmol) in tetrahydrofuran (5 mL) was added at - 40 °C, and the reactant was stirred at 25 °C for 2 h. The reactant was poured into water (20 mL). The organic phase was separated, and the aqueous phase was extracted 3 times with ethyl acetate (10 mL). The organic phase was washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by column chromatography on silica gel (petroleum ether / ethyl acetate = 1 / 0 - 1 / 2) to obtain Cpd 2 (0.5 g, yield 45%) as a colorless oil.

[0323] 1 H NMR (15017259 - 181 - P1 400 MHz, chloroform - d) δ 0.92 (d, J = 6.62 Hz, 6H), 1.50 (d, J = 6.62 Hz, 3H), 1.83 - 1.92 (m, 2H), 2.48 (d, J = 7.28 Hz, 2H), 4.88 (q, J = 6.39 Hz, 1H), 7.14 (d, J = 7.94 Hz, 2H), 7.29 (d, J = 7.94 Hz, 2H)

[0324] General procedure for the preparation of Cpd_2 - ET37412 - 153

Chem.

[0325] 1 H NMR (ET37412 - 153 - HNMRP1, 400 MHz, DMSO - d6) δ 0.85 (d, J = 6.63 Hz, 6H), 1.29 (d, J = 6.38 Hz, 3H), 1.73 - 1.85 (m, 1H), 2.41 (d, J = 7.13 Hz, 2H), 7.07 (d, J = 7.88 Hz, 2H), 7.23 (d, J = 7.88 Hz, 2H) General procedure for the preparation of Cpd 2 - ET37412 - 165

Chem.

[0326] General Procedure for the Preparation of Cpd_5a-ET37412-264

Chem.

[0327] 1 H NMR (ET37412-264-1, 400 MHz, CDCl3-d6) δ 0.08 - 0.14 (m, 9H), 2.19 (s, 3H), 4.23 (d, J = 1.47 Hz, 1H), 4.61 - 4.82 (m, 1H), 4.71 (d, J = 1.59 Hz, 1H), 6.97 (d, J = 8.07 Hz, 2H), 7.33 (d, J = 8.19 Hz, 2H)

[0328] General Procedure for the Preparation of Cpd 6a-NotebookPage:ET37412-267

Chem.

[0329] 11H NMR (ET37412 - 267 - 11, 400 MHz, CDCl3 - d6) δ 0.00 (s, 9H), 0.85 - 0.95 (m, 2H), 1.06 - 1.16 (m, 2H), 2.26 (s, 3H), 7.15 - 7.23 (m, 4H)

[0330] Cpd 3a - NotebookPage: General procedure for the preparation of ET37412 - 281

Chem.

[0331] 1 1H NMR (ET37412 - 281 - 2, 400 MHz, CDCl3 - d6) δ 0.95 - 1.05 (m, 2H), 1.18 - 1.25 (m, 2H), 2.34 (s, 3H), 7.11 - 7.25 (m, 4H)

[0332] General procedure for the preparation of Series 10 - K - P1 and Series 10 - K - P2 - ET37412 - 175

Chem.

[0333] 1 H NMR (ET37412-175-1, 400 MHz, MeOD) δ1.45 (d, J = 6.17 Hz, 3H), 3.76 (s, 3H), 5.43 (d, J = 6.39 Hz, 1H), 6.54 (s, 1H), 6.50 - 6.58 (m, 1H), 6.66 (d, J = 8.60 Hz, 1H), 7.10 (d, J = 8.60 Hz, 1H), 7.18 - 7.33 (m, 5H), 7.59 - 7.65 (m, 1H), 7.66 - 7.72 (m, 1H), 7.78 (s, 1H) LCMS (ESI+): m / z = 541.1 (M + H) + , RT: 3.187 minutes 5_95 AB_6 minutes - 220 - 254 - ELSD: LC / MS (gradient was 5% B at 0.40 minutes, 5 - 95% B from 0.40 to 3.00 minutes, held at 95% B for 1.00 minute, then 95 - 5% B in 0.01 minute, and the flow rate was 1.0 mL / min. Mobile phase A was 0.037% trifluoroacetic acid in water and mobile phase B was 0.018% trifluoroacetic acid in acetonitrile. The column used for chromatography was a Kinetex C18 50 × 2.1 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection and positive electrospray ionization. The MS range was 100 - 1000.)

[0334] SFC method: Column: Chiralcel OJ-3, 50×4.6mm I.D., 3μm Mobile phase: A: CO2 B: MeOH (0.05% IPAm, v / v) Gradient:

Table 3

[0335] SFC method: Column: Chiralcel OJ-3, 50×4.6mm I.D., 3μm Mobile phase: A: CO2 B: MeOH (0.05% IPAm, v / v) Gradient:

Table 4

Chem.

Chem.

Chem.

Chem.

[0336] General procedure for the preparation of Cpd 3 - ET37412 - 126

Chem.

[0337] General procedure for the preparation of Cpd_4 - ET37412 - 107

Chemical formula

[0338] LCMS method: LCMS (ESI+): m / z = 665.3 (M + H)+, RT: 0.996 min

[0339] 5 - 95 AB_2 min: The column used in LC / MS chromatography was Kinetex 5μm EVO C18 100A. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in HPLC grade acetonitrile. The gradient was 5 - 95% B in 2.20 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 1.00 min), 95 - 100% B (1.00 - 1.80 min), 5% B at 1.81 min, and held at 5% B for 0.39 min. The flow rate was 1.0 mL / min (0.01 - 1.80) 1.2 mL (1.81 - 2.20).

[0340] General procedure for the preparation of Cpd 5 - ET37412 - 140

Chem.

[0341] General procedure for the preparation of Cpd 7 - ET37412 - 141

Chem.

[0342] General procedure for the preparation of Cpd 8-ET37412-143 [Chemical formula] A solution of Cpd 7 (250 mg, 395.71 μmol) in HCl / dioxane (5 mL, 4 N) was stirred at 25 °C for 12 h. LCMS indicated the completion of the reaction. The reaction mixture was concentrated to give Cpd 8 (250 mg, 83.18% yield) as a colorless oil, which was used in the next step without purification.

[0343] LCMS (ESI+): m / z = 532.3 (M + H)+, RT: 0.637 min

[0344] 5 - 95 AB_2 min: The column used in LC / MS was Kinetex 5μm EVO C18 100A. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in HPLC grade acetonitrile. The gradient was 5 - 95% B in 2.20 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 1.00 min), 95 - 100% B (1.00 - 1.80 min), 5% B at 1.81 min, and held at 5% B for 0.39 min. The flow rate was 1.0 mL / min (0.01 - 1.80) 1.2 mL (1.81 - 2.20).

[0345] General procedure for the preparation of Series 10 - A - ET37412 - 147

Chem.

[0346] 1 H NMR: ET37412 - 147 - P1A1 (400 MHz, DMSO - d6) δ 0.88 - 0.97 (m, 9H), 1.74 (br t, J = 6.05 Hz, 2H), 1.90 (ddd, J = 12.81, 8.71, 4.52 Hz, 1H), 2.00 - 2.10 (m, 1H), 2.29 (br t, J = 6.36 Hz, 2H), 2.41 - 2.46 (m, 3H), 3.15 - 3.24 (m, 2H), 3.29 (br t, J = 6.17 Hz, 2H), 3.43 - 3.52 (m, 4H), 3.56 - 3.67 (m, 4H), 3.73 - 3.74 (m, 3H), 3.94 (br d, J = 1.47 Hz, 2H), 3.99 (br t, J = 5.99 Hz, 3H), 4.19 - 4.28 (m, 1H), 4.37 (br d, J = 6.60 Hz, 1H), 4.40 - 4.47 (m, 1H), 4.55 (d, J = 9.66 Hz, 1H), 6.71 (s, 1H), 6.76 (d, J = 8.68 Hz, 1H), 7.25 (d, J = 8.56 Hz, 1H), 7.39 (s, 4H), 7.44 (br d, J = 9.41 Hz, 1H), 7.64 (br d, J = 8.31 Hz, 1H), 7.75 (dd, J = 4.95, 3.24 Hz, 2H), 7.96 (br t, J = 5.50 Hz, 1H), 8.57 (t, J = 5.93 Hz, 1H), 8.97 (s, 1H) LCMS (ESI+): m / z = 1080.3 (M + H)+, RT: 2.723 minutes

[0347] 5_95AB_6 minutes_MS1500 - 220 - 254 - ELSD: LC / MS (Gradient was 5% B in 0.40 minutes, 5 - 95% B from 0.40 - 3.00 minutes, held at 95% B for 1.00 minute, then 95 - 5% B in 0.01 minute, and the flow rate was 1.0 mL / min. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in acetonitrile. The column used for chromatography was a Kinetex C18 50×2.1 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection and positive electrospray ionization. The MS range was 100 - 1500.)

[0348] Synthesis of Series 10 - B (NUCC - 0226304) [Chemical formula] Synthesis scheme: [Chemical formula] General procedure for the preparation of Cpd 2-ET37412-142 [Chemical formula] To a solution of Cpd 1 (215.76 mg, 868.87 mmol) in 1-methyl-2-pyrrolidinone (2 mL) was added N-ethyl-N-isopropylpropan-2-amine (187.16 mg, 1.45 mmol) and 2-(2,6-dioxo-3-piperidyl)-4-fluoro-isoindoline-1,3-dione (200 mg, 724.06 mmol). The reaction mixture was stirred at 90 °C for 12 h. TLC (petroleum ether / ethyl acetate = 1:1, Rf = 0.23) indicated the completion of the reaction. The reaction mixture was poured into water (20 mL), and the mixture was extracted with ethyl acetate (20 mL). The organic layer was concentrated to give a residue, which was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100 - 200 mesh silica gel, petroleum ether / ethyl acetate = 1 / 0, 0 / 1) to afford Cpd 2 (150 mg, yield 41%) as a colorless oil.

[0349] General procedure for the preparation of Cpd 3-ET37412-144 [Chemical formula] A solution of Cpd 2 (200 mg, 396.41 mmol) in HCl / dioxane (2 mL, 4 M) was stirred at 25 °C for 0.2 h. LCMS indicated the completion of the reaction. The reaction mixture was concentrated to afford Cpd 3 (120 mg, yield 75%) as a colorless oil, which was used in the next step without purification.

[0350] LCMS method: LCMS (ESI+): m / z = 405.1 (M + H)+, RT: 0.605 min

[0351] 5 - 95 AB_2 min: The column used in LC / MS was Kinetex 5μm EVO C18 100A. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in HPLC grade acetonitrile. The gradient was 5 - 95% B in 2.20 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 1.00 min), 95 - 100% B (1.00 - 1.80 min), 5% B at 1.81 min, and held at 5% B for 0.39 min. The flow rate was 1.0 mL / min (0.01 - 1.80) 1.2 mL (1.81 - 2.20).

[0352] General procedure for the preparation of Cpd_4 - ET37412 - 158

Chemical formula

[0353] 1 H NMR: ET37412 - 158 - g1 (400 MHz, DMSO - d6) δ 1.71 - 1.76 (m, 3H), 1.76 - 1.84 (m, 2H), 1.94 - 2.06 (m, 1H), 2.27 (t, J = 6.36 Hz, 2H), 2.53 - 2.65 (m, 3H), 2.82 - 2.94 (m, 1H), 3.16 (q, J = 5.71 Hz, 2H), 3.28 - 3.39 (m, 6H), 3.53 - 3.63 (m, 6H), 3.74 - 3.83 (m, 3H), 4.10 (br t, J = 5.99 Hz, 2H), 5.05 (dd, J = 12.90, 5.44 Hz, 1H), 6.59 (br s, 1H), 6.68 (s, 1H), 7.03 (d, J = 7.09 Hz, 1H), 7.13 (d, J = 8.56 Hz, 1H), 7.23 (d, J = 8.68 Hz, 1H), 7.54 - 7.64 (m, 3H), 7.72 (d, J = 1.47 Hz, 1H), 7.79 - 7.88 (m, 2H), 11.08 (s, 1H) General procedure for the preparation of Series 10 - B - ET37412 - 162

Chemical Structure

[0354] 1 H NMR: ET37412 - 162 - 1 (400 MHz, CDCl3 - d6) δ 1.87 - 1.96 (m, 2H), 2.08 - 2.19 (m, 1H), 2.44 - 2.57 (m, 2H), 2.68 - 2.77 (m, 2H), 2.82 - 2.91 (m, 1H), 3.41 - 3.48 (m, 6H), 3.55 - 3.67 (m, 9H), 3.68 - 3.74 (m, 3H), 3.84 (s, 3H), 4.04 (br t, J = 6.02 Hz, 2H), 4.89 (br dd, J = 12.04, 5.36 Hz, 1H), 6.57 (s, 1H), 6.69 (d, J = 8.58 Hz, 1H), 6.90 (br d, J = 8.46 Hz, 2H), 7.09 - 7.14 (m, 1H), 7.21 (d, J = 8.46 Hz, 1H), 7.47 - 7.54 (m, 2H), 7.60 - 7.64 (m, 1H), 7.75 (s, 1H), 8.63 (br s, 1H) LCMS (ESI+): m / z = 953.3 (M + H) + , RT: 2.831 minutes

[0355] 5_95AB_6 minutes_MS1500 - 220 - 254 - ELSD: LC / MS (Gradient was 5% B in 0.40 minutes, 5 - 95% B from 0.40 to 3.00 minutes, held at 95% B for 1.00 minute, then 95 - 5% B in 0.01 minute, and the flow rate was 1.0 mL / min. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in acetonitrile. The column used for chromatography was a Kinetex C18 50×2.1 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection and positive electrospray ionization. The MS range was 100 - 1500.)

[0356] Final Report of NUCC - 0226504 (ET42365 - 21 - 1), NUCC - 0226503 (ET42365 - 19 - 1), NUCC - 0226499 (ET42365 - 26 - 1), NUCC - 0226498 (ET42365 - 25 - 1), NUCC - 0226497 (ET42365 - 23 - 1) and NUCC - 0226496 (ET42365 - 20 - 1)

Chem.

Chem.

[0357]

Chem.

[0358] 50_100 CD_6 min - 220 - 254 - ELSD: LC / MS (Gradient: 50% B in 0.40 min, 50 - 100% B from 0.40 - 3.40 min, hold at 100% B for 0.45 min, then 100 - 50% B in 0.01 min, flow rate 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3, mobile phase B was acetonitrile. The column used for chromatography was an Xbridge C18 2.1×50 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0359] General procedure for the preparation of compound 4 - ET42365 - 6 [Chemical formula] To a solution of compound 3 (2.5 g, 7.02 mmol, 1 equiv) in DMF (50 mL) was added K2CO3 (1.94 g, 14.04 mmol, 2 equiv) and 1 - (bromomethyl) - 4 - chloro - benzene (1.73 g, 8.43 mmol, 1.2 equiv), and the reaction was stirred at 30 °C for 8 h. LCMS indicated that the starting material was consumed and a new peak with the desired product Ms was detected. The reaction was diluted with water (150 mL), the precipitate was filtered off and dried under high vacuum to give compound 4 (2 g, 56.3% yield) as a white solid. The product was used directly in the next step without further purification.

[0360] LCMS (ESI+): RT: 1.144 min, m / z = 480.8 (M + H)

[0361] 5 - 95AB_2 min: The column used in LC / MS was an Agilent Poroshell SB - C18 3.0×30 mm, 2.7 μm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in HPLC - grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B (0.70 - 1.16 min), 95 - 5% B (1.16 - 1.50 min). The flow rate was 1.5 mL / min.

[0362] General procedure for the preparation of compound 5 - ET42365 - 10

Chemical formula

[0363] LCMS(ESI+): RT: 0.936 min, m / z = 499.1 (M + H) +

[0364] 5 - 95 AB_2 min: The column used in LC / MS chromatography was Agilent Poroshell SB - C18 3.0×30 mm, 2.7 μm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in HPLC - grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B (0.70 - 1.16 min), 95 - 5% B (1.16 - 1.50 min). The flow rate was 1.5 mL / min.

[0365] General procedure for the preparation of A01B03C01D01 - ET42365 - 20

Chem.

[0366] 11H NMR: (ET42365 - 20 - P1X, 400 MHz, chloroform - d) δ 3.84 (s, 3H), 5.07 (s, 2H), 5.15 (s, 1H), 6.55 (s, 1H), 6.70 (d, J = 8.51 Hz, 1H), 7.19 (t, J = 8.88 Hz, 3H), 7.26 (d, J = 1.88 Hz, 1H), 7.33 (d, J = 8.38 Hz, 2H), 7.55 (d, J = 1.88 Hz, 1H), 7.59 (d, J = 8.25 Hz, 1H) LCMS(ESI+): RT: 2.889 min, m / z = 527.1, 529.0 (M + H, M + 2 + H) +

[0367] 50_100 CD_6 min - 220 - 254 - ELSD: LC / MS (The gradient was 50% B at 0.40 min, 50 - 100% B from 0.40 to 3.40 min, hold at 100% B for 0.45 min, then 100 - 50% B in 0.01 min, and the flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3 and mobile phase B was acetonitrile. The column used for chromatography was an Xbridge C18 2.1×50 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0368] Synthesis of Series 10 - C (NUCC - 0226502)

Chemical Structure

Chemical Structure

Chemical Structure

[0369] LCMS method: LCMS (ESI+): m / z = 363.1 (M+H)+, RT: 0.837 min

[0370] 5 - 95 AB_2 min: LC / MS (The column used for chromatography was Kinetex 5μm EVO C18 100A. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in HPLC grade acetonitrile. The gradient was 5 - 95% B in 2.20 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 1.00 min), 95 - 100% B (1.00 - 1.80 min), 5% B at 1.81 min, and held at 5% B for 0.39 min. The flow rate was 1.0 mL / min (0.01 - 1.80) 1.2 mL (1.81 - 2.20).

[0371] General procedure for the preparation of Cpd 3 - ET37412 - 183

Chemical formula

[0372] 1 H NMR: ET37412-183-1 (400 MHz, CDCl3) δ 5.00 (br s, 1H), 5.65 (br d, J = 3.30 Hz, 1H), 6.39 (d, J = 2.20 Hz, 1H), 6.57 (s, 1H), 6.71 (dd, J = 8.80, 2.20 Hz, 1H), 7.31 - 7.41 (m, 10H), 7.94 (d, J = 8.80 Hz, 1H) General procedure for the preparation of Cpd_4-ET37412-214

Chemical formula

[0373] LCMS (ESI+): m / z = 476.2 (M + H) + , RT: 0.935 min

[0374] General procedure for the preparation of Cpd 5 NotebookPage: ET37412-234

[0375]

Chemical formula

[0376] LCMS: m / z = 574.4 (M+H) + , room temperature: 1.211 min 5-95AB_2 min: LC / MS (the column used for chromatography was Kinetex 5 μm EVO C18 100A 2.1×30 mm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water and mobile phase B was 0.02% trifluoroacetic acid in HPLC grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B for 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B for 0.46 min, 95 - 5% B (1.61 - 1.50 min), and held at 5% B for 0.11 min. The flow rate was 1.5 mL / min.

[0377] General procedure for the preparation of Series 10-C NotebookPage: ET37412-246

[0378]

Chemical Structure

[0379] 1 H NMR (ET37412-246-P1A5, 400 MHz, DMSO-d6) δ 3.71 (s, 3H), 4.86 (d, J = 5.95 Hz, 1H), 5.67 (d, J = 5.73 Hz, 1H), 6.27 (d, J = 8.60 Hz, 1H), 6.60 (s, 1H), 6.99 (d, J = 8.38 Hz, 1H), 7.22 (td, J = 5.68, 2.54 Hz, 2H), 7.26 - 7.34 (m, 8H), 7.72 (dd, J = 8.38, 1.54 Hz, 1H), 7.81 (dd, J = 4.85, 3.31 Hz, 2H), 8.53 (s, 1H) LCMS: m / z = 602.2 (M + H) + , room temperature: 3.023 min 5_95AB_6 min - 220 - 254 - ELSD: LC / MS (The gradient was 5% B from 0.40 min, 5 - 95% B from 0.40 to 3.00 min, 95% B was held for 1.00 min, then 95 - 5% B in 0.01 min, and the flow rate was 1.0 mL / min. Mobile phase A was 0.037% trifluoroacetic acid in water, and mobile phase B was 0.018% trifluoroacetic acid in acetonitrile. The column used for chromatography was a Kinetex C18 50×2.1 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection and positive electrospray ionization. The MS range was 100 - 1000.)

[0380] Synthesis of NUCC - 0226595 (ET42365 - 226 - 1) [Chemical formula] Synthesis scheme: Preparation of A03B01D01 (NUCC - 0226595) [Chemical formula] Chemical synthesis 5 - 95AB_2 min: LC / MS (The column used for chromatography was an Agilent Poroshell SB - C18 3.0×30 mm, 2.7 μm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in HPLC - grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B (0.70 - 1.16 min), 95 - 5% B (1.16 - 1.50 min). The flow rate was 1.5 mL / min.)

[0381] 5_95AB_6 min - 220 - 254 - ELSD: LC / MS (Gradient: 5% B from 0.40 min, 5 - 95% B from 0.40 - 3.00 min, hold at 95% B for 1.00 min, then 95 - 5% B in 0.01 min, flow rate was 1.0 mL / min. Mobile phase A was 0.037% trifluoroacetic acid in water, and mobile phase B was 0.018% trifluoroacetic acid in acetonitrile. The column used for chromatography was a Kinetex C18 50×2.1 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection and positive electrospray ionization. The MS range was 100 - 1000.)

[0382] Experiment for the largest - scale execution: General procedure for the preparation of compound 2 - ET43365 - 193

[0383] [Chemical formula] To a solution of compound 1 (1 g, 5.75 mmol, 1 equiv), [4 - chloro - 3 - (trifluoromethyl)phenyl] - boronic acid (1.94 g, 8.63 mmol, 1.5 equiv) and Na2CO3 (1.22 g, 11.50 mmol, 2 equiv) in a mixed solution of toluene (30 mL), EtOH (6 mL) and H2O (1.2 mL), Pd(dppf)Cl2·CH2Cl2 (469.35 mg, 575.00 μmol, 0.1 equiv) was added under nitrogen, and the reaction mixture was stirred at 100 °C for 2 h. LCMS indicated that the starting material was consumed and a new peak with the desired product Ms was detected. The reaction mixture was filtered and the filtrate was concentrated to obtain the crude product. The crude product was diluted with water (60 mL) and extracted with ethyl acetate (3×50 mL). The organic layer was washed with aqueous K2CO3 solution (2×60 mL) and brine (60 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 2 (0.5 g, yield 28.6%) as a yellow solid. The crude product was used directly in the next step without further purification.)

[0384] 11H NMR (ET42365-193-P1A, 400 MHz, chloroform-d) δ 7.21 - 7.25 (m, 1H), 7.28 - 7.31 (m, 1H), 7.60 (d, J = 8.25 Hz, 1H), 8.08 (dd, J = 8.32, 1.69 Hz, 1H), 8.33 (d, J = 2.25 Hz, 2H)

[0385] General procedure for the preparation of compound 3 - ET42365-207 [Chemical formula] To a solution of compound 2 (400 mg, 1.46 mmol, 1 eq) in DMF (4 mL), NBS (312.21 mg, 1.75 mmol, 1.2 eq) was added portionwise at 0 °C, and the reaction mixture was stirred at 20 °C for 3 h. LCMS indicated that approximately 30% of the starting material remained, and a new peak (approx. 35%) with the desired product Ms was detected. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (3 × 15 mL). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, and concentrated to give the crude product. The crude product was purified by Prep-HPLC (column: Phenomenex Gemini-NX 80×40 mm×3 μm; mobile phase: [water (10 mM NH4HCO3) - ACN]; B%: 30% - 60%, 8 min) to afford compound 3 (120 mg, yield 23.29%) as a yellow solid.

[0386] LCMS (ESI+): RT = 0.833 min, m / z 352.0, 354.0 (M + H) +

[0387] 5 - 95 AB_2 min: The column used in LC / MS was an Agilent Poroshell SB - C18 3.0×30 mm, 2.7 μm. The detection method was a diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in HPLC - grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B (0.70 - 1.16 min), 95 - 5% B (1.16 - 1.50 min). The flow rate was 1.5 mL / min.

[0388] General procedure for the preparation of A03B01D01 - ET42365 - 226

[0389] [Chemical formula] To a solution of compound 3 (120 mg, 340.39 μmol, 1 equiv), [2 - methyl - 5 - (trifluoromethyl) - pyrazol - 3 - yl]boronic acid (85.81 mg, 442.51 μmol, 1.3 equiv) and K3PO4 (144.51 mg, 680.79 μmol, 2 equiv) in a mixed solution of H2O (1 mL) and THF (5 mL), di - tert - butyl(cyclopentyl)phosphane; dichloropalladium; iron (22.18 mg, 34.04 μmol, 0.1 equiv) was added under a nitrogen atmosphere, and the reaction mixture was stirred at 80 °C for 12 h. LCMS indicated that the starting material was consumed and a new peak with the desired product Ms was detected. The reaction mixture was filtered and the filtrate was concentrated to obtain a crude product. The crude product was purified by Prep - HPLC (column: Waters Xbridge preparative OBD C18 150×40 mm×10 μm; mobile phase: [water (10 mM NH4HCO3) - ACN]; B%: 65% - 95%, 8 min) to obtain A03B01D01 (26.8 mg, yield 18.37%) as a white solid.

[0390] 11H NMR (ET42365-257-P1A, 400 MHz, chloroform-d) δ 4.28 (s, 3H), 5.50 (br s, 1H), 6.81 (s, 1H), 7.36 (d, J = 8.38 Hz, 1H), 7.53 (d, J = 8.38 Hz, 1H), 7.65 (d, J = 8.50 Hz, 1H), 8.14 - 8.21 (m, 1H), 8.43 (s, 1H) LCMS (ESI+): RT = 3.535 min, m / z 422.1 (M + H) +

[0391] 5_95AB_6 min - 220 - 254 - ELSD: LC / MS (Gradient was 5% B in 0.40 min, 5 - 95% B from 0.40 to 3.00 min, hold 95% B for 1.00 min, then 95 - 5% B in 0.01 min, flow rate was 1.0 mL / min. Mobile phase A was 0.037% trifluoroacetic acid in water and mobile phase B was 0.018% trifluoroacetic acid in acetonitrile. The column used for chromatography was a Kinetex C18 50×2.1 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection and positive electrospray ionization. The MS range was 100 - 1000.)

[0392] Synthesis of NUCC - 0226574 (ET42365 - 172 - 1)

Chemical Structure

Chemical Structure

[0393] 50_100 AB_6 min - 220 - 254 - ELSD: LC / MS (The gradient was 50% B in 0.40 min, 50 - 100% B from 0.40 to 3.00 min, hold at 100% B for 1.00 min, then 100 - 50% B in 0.01 min, and the flow rate was 1.0 mL / min. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in acetonitrile. The column used for chromatography was a Luna C18 50×2.0 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection as well as positive electrospray ionization. The MS range was 100 - 1000.)

[0394] Experiment for the largest - scale execution: General procedure for the preparation of compound 7 - ET43365 - 132

[0395] [Chemical formula] A solution of core A3_1 (600 mg, 1.25 mmol, 1 equiv) and pyridine (396.51 mg, 5.01 mmol, 404.61 μL, 4 equiv) in DCM (20 mL) was added dropwise with Tf2O (530.37 mg, 1.88 mmol, 310.16 μL, 1.5 equiv) at 0 °C, and the reaction mixture was stirred at 20 °C for 12 h. TLC (petroleum ether / ethyl acetate = 2 / 1) indicated that the starting material was consumed and a new peak was formed. The reaction mixture was diluted with water (60 mL) and extracted with DCM (3 × 30 mL). The organic layers were separated, combined organic layers were washed with brine (60 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography on silica gel (eluting with petroleum ether / ethyl acetate = 50 / 1 to 10 / 1) to give compound 7 (600 mg, yield 74.46%) as a white solid.

[0396] 1 H NMR (ET42365-132-P1A, 400 MHz, chloroform-d) δ 1.80 (s, 3H), 3.85 (s, 3H), 6.57 (s, 1H), 7.47 (dd, J = 8.25, 1.75 Hz, 1H), 7.49 - 7.56 (m, 2H), 7.64 (d, J = 8.25 Hz, 1H), 7.69 (d, J = 1.63 Hz, 1H)

[0397] General procedure for the preparation of compound 12A-ET42365-142

Chemical Structure

[0398] 1 H NMR (ET42365 - 142 - P1A, 400 MHz, chloroform - d) δ 1.22 (s, 12H), 6.04 (d, J = 18.39 Hz, 1H), 7.16 - 7.24 (m, 3H), 7.29 - 7.36 (m, 2H)

[0399] General procedure for the preparation of Compound 12 - ET42365 - 151

Chemical Structure

[0400] LCMS (ESI+): RT = 0.971 min, m / z 599.2 (M+H) +

[0401] 5-95AB_2 min: LC / MS (The column used for chromatography was Agilent Poroshell SB-C18 3.0×30 mm, 2.7 μm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in HPLC grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B (0.70 - 1.16 min), 95 - 5% B (1.16 - 1.50 min). The flow rate was 1.5 mL / min.

[0402] General procedure for the preparation of Compound 13-ET42365-154

[0403]

Chemical Structure

[0404] LCMS (ESI+): RT = 0.969 min, m / z 557.2 (M + H) +

[0405] 5 - 95 AB_2 min: The column used for LC / MS was an Agilent Poroshell SB - C18 3.0 × 30 mm, 2.7 μm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in HPLC - grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B (0.70 - 1.16 min), 95 - 5% B (1.16 - 1.50 min). The flow rate was 1.5 mL / min.

[0406] General procedure for the preparation of compound 13 - ET42365 - 154

[0407]

Chemical Structure

[0408] 1 H NMR (ET42365-172-P1A, 400 MHz, chloroform-d) δ 2.65 - 2.79 (m, 4H), 3.86 (s, 3H), 4.82 (s, 1H), 6.61 (s, 1H), 6.83 (d, J = 8.38 Hz, 2H), 7.03 (d, J = 7.88 Hz, 1H), 7.17 - 7.21 (m, 2H), 7.22 - 7.26 (m, 2H), 7.52 (d, J = 2.00 Hz, 1H), 7.63 (d, J = 8.13 Hz, 1H) LCMS (ESI+): RT = 3.135 min, m / z 559.1 (M + H) +

[0409] 50_100AB_6 min - 220 - 254 - ELSD: LC / MS (gradient was 50% B at 0.40 min, 50 - 100% B from 0.40 - 3.00 min, hold at 100% B for 1.00 min, then 100 - 50% B in 0.01 min, flow rate was 1.0 mL / min. Mobile phase A was 0.04% trifluoroacetic acid in water and mobile phase B was 0.02% trifluoroacetic acid in acetonitrile. The column used for chromatography was a Luna C18 50×2.0 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection and positive electrospray ionization. The MS range was 100 - 1000.)

[0410] Synthesis of NUCC-0226566 (ET42365-147-1)

Chem.

Chem.

[0411] 50_100 CD_6 min - 220 - 254 - ELSD: LC / MS (The gradient was 50% B in 0.40 min, 50 - 100% B from 0.40 - 3.40 min, hold at 100% B for 0.45 min, then 100 - 50% B in 0.01 min, and the flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3, and mobile phase B was acetonitrile. The column used for chromatography was an Xbridge C18 2.1×50 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0412] Experiment for the largest scale execution: General procedure for the preparation of compound 10 - ET43365 - 145

[0413]

Chem.

[0414] LCMS (ESI+): RT = 0.959 min, m / z 587.2 (M+H) +

[0415] 5-95 AB_2 min: LC / MS (The column used for chromatography was Agilent Poroshell SB-C18 3.0×30 mm, 2.7 μm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in HPLC grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B (0.70 - 1.16 min), 95 - 5% B (1.16 - 1.50 min). The flow rate was 1.5 mL / min.

[0416] General procedure for the preparation of A01B01C08D01-ET42365-147

[0417] [Chemical formula] To a solution of compound 10 (120 mg, approximately 163.45 μmol, crude) in MeOH (5 mL), K2CO3 (45.18 mg, 326.90 μmol, 2 equivalents) was added, and the reaction mixture was stirred at 20 °C for 3 hours. LCMS indicated that the starting material was consumed and a new peak with the desired product Ms was detected. The reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (3 × 30 mL). The organic layers were separated, combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by Prep-HPLC (column: Waters Xbridge preparative OBD C18 150×40 mm×10 μm; mobile phase: [water (10 mM NH4HCO3)-ACN]; B%: 70% - 95%, 8 minutes) to obtain A01B01C08D01 (62.8 mg, yield 70.46%) as an off-white solid.

[0418] 1 H NMR (ET42365-147-P1A, 400 MHz, chloroform-d) δ 3.73 (d, J = 3.13 Hz, 2H), 3.87 (s, 3H), 4.86 (br s, 1H), 6.61 (s, 1H), 6.80 (d, J = 8.25 Hz, 2H), 6.99 (d, J = 7.88 Hz, 1H), 7.19 (d, J = 8.38 Hz, 2H), 7.25 (s, 1H), 7.30 (br d, J = 1.50 Hz, 1H), 7.41 (d, J = 1.50 Hz, 1H), 7.60 (d, J = 8.25 Hz, 1H) LCMS (ESI+): RT = 3.026 minutes, m / z 545.1 (M + H) +

[0419] 50_100CD_6 minutes - 220 - 254 - ELSD:LC / MS (Gradient: 50%B in 0.40 minutes, 50 - 100%B from 0.40 to 3.40 minutes, hold at 100%B for 0.45 minutes, then 100 - 50%B in 0.01 minute, flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3, and mobile phase B was acetonitrile. The column used for chromatography was an Xbridge C18 2.1×50 mm column (5μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0420] Synthesis of NUCC - 0226548 (ET42365 - 124 - 1)

Chemical Structure

Chemical Structure

Chemical Structure

[0421] NEG50_100CD_6 min - 220 - 254 - ELSD: LC / MS (Gradient: 50% B at 0.40 min, 50 - 100% B from 0.40 to 3.40 min, hold at 100% B for 0.45 min, then 100 - 50% B in 0.01 min, flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3 and mobile phase B was acetonitrile. The column used for chromatography was an Xbridge C18 2.1×50 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection. The MS mode was negative electrospray ionization. The MS range was 100 - 1000.)

[0422] Experiment for the largest - scale execution: General procedure for the preparation of compound 2 - ET43365 - 97

[0423] [Chemical formula] To a solution of compound 1 (5 g, 47.56 mmol, 4.76 mL, 1 equiv) in toluene (60 mL) was added isobenzofuran - 1,3 - dione (7.04 g, 47.56 mmol, 1 equiv). The mixture was stirred at 120 °C for 12 h. LCMS indicated that the starting material was consumed and a new peak with the desired product Ms was detected. The reaction mixture was concentrated to give compound 2 (11 g, yield 88.49%) as a yellow solid. The product was used directly in the next step without further purification.

[0424] 1 H NMR (ET42365 - 97 - P1A, 400 MHz, chloroform - d) δ 2.66 (br s, 1H), 3.59 - 3.63 (m, 2H), 3.67 - 3.71 (m, 2H), 3.73 - 3.78 (m, 2H), 3.89 - 3.95 (m, 2H), 7.70 - 7.76 (m, 2H), 7.83 - 7.89 (m, 2H)

[0425] General procedure for the preparation of compound 3 - ET42365 - 103 [Chemical formula] To a solution of compound 2 (3 g, 12.75 mmol, 1 equiv) and Et3N (3.23 g, 31.88 mmol, 4.44 mL, 2.5 equiv) in DCM (60 mL) was added dropwise MsCl (2.19 g, 19.13 mmol, 1.48 mL, 1.5 equiv) at 0 °C, and the reaction mixture was stirred at 20 °C for 12 h. LCMS indicated that the starting material was consumed and a new peak with the desired product Ms was detected. The reaction mixture was diluted with water (100 mL) and extracted with DCM (2 × 100 mL). The organic layers were separated, combined organic layers were washed with brine (200 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography on silica gel (petroleum ether / ethyl acetate = 20 / 1 to 2 / 1) to give compound 3 (3.7 g, yield 87.97%) as a yellow solid.

[0426] 1 H NMR (ET42365 - 103 - P1A, 400 MHz, chloroform - d) δ 3.00 (s, 3H), 3.71 - 3.82 (m, 4H), 3.88 - 3.95 (m, 2H), 4.28 - 4.36 (m, 2H), 7.70 - 7.78 (m, 2H), 7.82 - 7.91 (m, 2H)

[0427] General procedure for the preparation of compound 4 - ET43587 - 64

Chemical formula

[0428] LCMS (ESI+): RT = 0.734 min, m / z 346.0 (M+H) +

[0429] 5-95AB_2 min: LC / MS (The column used for chromatography was Kinetex EVO C18 2.1×30 mm, 5 μm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water and mobile phase B was 0.02% trifluoroacetic acid in HPLC grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B (0.70 - 1.16 min), 95 - 5% B (1.16 - 1.50 min). The flow rate was 1.5 mL / min.

[0430] General procedure for the preparation of compound BC1_Pht-ET42365-118

[0431]

Chemical Structure

[0432] LCMS (ESI+): RT = 0.656 min, m / z 324.2 (M + H) +

[0433] 5 - 95 AB_2 min: LC / MS (The column used for chromatography was Kinetex EVO C18 2.1×30 mm, 5 μm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water and mobile phase B was 0.02% trifluoroacetic acid in HPLC grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B (0.70 - 1.16 min), 95 - 5% B (1.16 - 1.50 min). The flow rate was 1.5 mL / min.

[0434] General procedure for the preparation of A4BC1_2 - Pht - ET42365 - 119

[0435]

Chemical Structure

[0436] LCMS (ESI+): RT = 0.951 min, m / z 824.3 (M+H) +

[0437] 5 - 95 AB_2 min: LC / MS (The column used for chromatography was Kinetex EVO C18 2.1×30 mm, 5 μm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in HPLC grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B (0.70 - 1.16 min), 95 - 5% B (1.16 - 1.50 min). The flow rate was 1.5 mL / min.

[0438] General procedure for the preparation of compound 7 - ET42365 - 114

[0439]

Chemical Structure

[0440] 1 H NMR (ET42365-114-P1A, 400 MHz, chloroform-d) δ 2.05 - 2.21 (m, 1H), 2.71 - 2.90 (m, 2H), 2.93 - 3.08 (m, 1H), 3.22 (s, 3H), 4.99 (br dd, J = 12.67, 5.40 Hz, 1H), 7.43 (t, J = 8.41 Hz, 1H), 7.69 - 7.74 (m, 1H), 7.74 - 7.82 (m, 1H)

[0441] General procedure for the preparation of A4BC1_NH2-ET43259-123

Chemical formula

[0442] LCMS (ESI+): RT = 0.736 min, m / z 630.3 (M+H) +

[0443] 5 - 95 AB_2 min: LC / MS (The column used for chromatography was Kinetex EVO C18 2.1×30 mm, 5 μm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in HPLC - grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B (0.70 - 1.16 min), 95 - 5% B (1.16 - 1.50 min). The flow rate was 1.5 mL / min.)

[0444] General procedure for the preparation of A4BC1R1A - ET42365 - 124

Chemical Structure

[0445]

Chemical Structure

[0446] 1 1H NMR (ET42365-124-P1A, 400 MHz, chloroform-d) δ 2.04 - 2.13 (m, 1H), 2.68 - 2.83 (m, 2H), 2.90 - 3.04 (m, 1H), 3.21 (s, 3H), 3.51 (t, J = 5.38 Hz, 2H), 3.80 (t, J = 5.44 Hz, 2H), 3.83 - 3.90 (m, 5H), 4.10 - 4.18 (m, 2H), 4.86 - 4.95 (m, 1H), 5.02 (s, 2H), 5.11 (s, 1H), 6.57 (s, 1H), 6.78 (d, J = 8.63 Hz, 1H), 6.88 (d, J = 8.76 Hz, 2H), 6.93 (d, J = 8.50 Hz, 1H), 7.09 (d, J = 7.13 Hz, 1H), 7.15 (d, J = 8.63 Hz, 2H), 7.21 (d, J = 8.51 Hz, 1H), 7.47 (dd, J = 8.51, 7.25 Hz, 1H), 7.51 - 7.55 (m, 1H), 7.57 - 7.63 (m, 1H), 7.79 (d, J = 1.75 Hz, 1H) LCMS (ESI+): RT = 2.569 min, m / z 898.2 (M - H) +

[0447] NEG50_100CD_6 min - 220 - 254 - ELSD: LC / MS (Gradient was 50% B in 0.40 min, 50 - 100% B from 0.40 - 3.40 min, hold at 100% B for 0.45 min, then 100 - 50% B in 0.01 min, and the flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3 and mobile phase B was acetonitrile. The column used for chromatography was an Xbridge C18 2.1×50 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection. The MS mode was negative electrospray ionization. The MS range was 100 - 1000.)

[0448] Synthesis of Series A5 - 3B (NUCC - 0226604) and Series A5 - 3C (NUCC - 0226603)

Chemical Structure

[0449] Method 2: 5_95CD_6 min - 220 - 254 - ELSD LC / MS (Gradient was 5% B in 0.40 min, 5 - 95% B from 0.40 to 3.40 min, held at 95% B for 0.45 min, then 95 - 5% B in 0.01 min, and the flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3, and mobile phase B was acetonitrile. The column used for chromatography was an Xbridge Shield RP18 2.1×50 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection as well as positive electrospray ionization. The MS range was 100 - 1000.)

[0450] Experiment for the largest - scale execution: General procedure for the preparation of C - 4A - ET43318 - 125

[0451] [Chem.] To a mixture of C-4B (2 g, 11.29 mmol, 1.82 mL) and HCl (12 M, 940.89 μL, 1 equiv) in EtOH (30 mL), PtO2 (256.38 mg, 1.13 mmol) was added at 20 °C. The mixture was stirred under hydrogen at a pressure of 15 psi for 12 h. TLC (ethyl acetate / MeOH = 5 / 1) indicated that the starting material was consumed and a new spot appeared. The product having the desired Ms was detected by LCMS. The mixture was filtered through celite and the filtrate was concentrated under reduced pressure to give C-4A (2.2 g, yield 71.63%) as a yellow solid, which was used directly in the next step without further purification.

[0452] 1 H NMR (400 MHz, chloroform-d) δ 1.34 (t, J = 7.07 Hz, 6H), 2.41 (dt, J = 18.51, 7.75 Hz, 2H), 3.15 - 3.38 (m, 2H), 4.03 - 4.26 (m, 4H), 7.70 - 8.72 (m, 2H)

[0453] General procedure for the preparation of compound 3-1-ET43318-157

Chemical formula

[0454] LCMS indicated that 12.9% of the starting material remained and a 45.5% peak (Rt = 0.893 min) having the desired Ms was detected. The reaction mixture was poured into water (100 mL) and extracted with ethyl acetate (30 mL × 3). The organic layers were combined, washed with brine (2 × 10 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (eluting with petroleum ether / ethyl acetate = 20 / 1 - 3 / 1) to give 3-1 (250 mg, yield 24.06%) as a white solid.

[0455] 1 1H NMR (400 MHz, chloroform-d) δ 1.79 (s, 3H), 3.82 (s, 3H), 5.22 (s, 2H), 6.51 (s, 1H), 6.79 (d, J = 8.00 Hz, 1H), 7.06 (d, J = 8.63 Hz, 1H), 7.32 - 7.36 (m, 1H), 7.38 - 7.43 (m, 3H), 7.49 (dd, J = 8.25, 1.75 Hz, 1H), 7.57 (br d, J = 8.50 Hz, 1H), 7.68 (d, J = 8.13 Hz, 1H), 7.76 (d, J = 1.75 Hz, 1H), 10.02 (s, 1H)

[0456] General procedure for the preparation of Compound 3 - 2A - ET43318 - 165

Chemical Structure

[0457] 11H NMR (400 MHz, chloroform-d) δ 1.31 (t, J = 7.07 Hz, 6H), 1.61 (br s, 4H), 1.78 (s, 3H), 2.00 (dt, J = 18.17, 7.18 Hz, 2H), 2.87 - 2.99 (m, 2H), 3.75 - 3.84 (m, 5H), 4.05 - 4.16 (m, 4H), 5.12 (s, 2H), 6.50 (s, 1H), 7.07 (d, J = 8.63 Hz, 1H), 7.20 (d, J = 8.00 Hz, 2H), 7.28 - 7.35 (m, 3H), 7.44 - 7.50 (m, 1H), 7.51 - 7.57 (m, 1H), 7.73 (d, J = 1.75 Hz, 1H)

[0458] General procedure for the preparation of compound series A5 - 3B - ET43318 - 168 [Chemical formula] To a solution of 3 - 2A (80 mg, 104.98 μmol) in MeOH (1 mL), K2CO3 (43.53 mg, 314.93 μmol) was added at 20 °C. The mixture was stirred for 12 h. LCMS indicated that the starting material was consumed and an 86% peak with the desired Ms (Rt = 0.737 min) was detected. The mixture was filtered to obtain a filtrate. The filtrate was purified by prep - HPLC to give series A5 - 3B (30.6 mg, yield 40.48%) as an off - white solid.

[0459] Method of prep - HPLC: Equipment: Gilson 281 semi - preparative HPLC system Mobile phase: A: 10 mM NH4HCO3 in H2O; B: acetonitrile Column: Waters Xbridge BEH C18 100×30 mm×10 μm Flow rate: 25 mL / min Monitoring wavelength: 220 and 254 nm [Table 5] 11H NMR (400 MHz, chloroform-d) δ 1.30 (t, J = 7.03 Hz, 6H), 1.95 (t, J = 7.15 Hz, 1H), 1.99 (t, J = 7.15 Hz, 1H), 2.89 (dt, J = 15.83, 7.18 Hz, 2H), 3.76 (s, 2H), 3.84 (s, 3H), 4.01 - 4.13 (m, 4H), 5.08 (s, 2H), 6.56 (s, 1H), 6.75 (d, J = 8.68 Hz, 1H), 7.19 (t, J = 7.83 Hz, 3H), 7.27 - 7.30 (m, 2H), 7.51 - 7.56 (m, 1H), 7.57 - 7.63 (m, 1H), 7.80 (d, J = 1.83 Hz, 1H) LCMS (ESI+): RT = 2.476 min, m / z 720.1 (M + H) + 5_95AB_6 min - 220 - 254 - ELSD

[0460] 5_95AB_6 min - 220 - 254 - ELSD: LC / MS (gradient was 5% B in 0.40 min, 5 - 95% B from 0.40 - 3.00 min, held at 95% B for 1.00 min, then 95 - 5% B in 0.01 min, and the flow rate was 1.0 mL / min. Mobile phase A was 0.037% trifluoroacetic acid in water and mobile phase B was 0.018% trifluoroacetic acid in acetonitrile. The column used for chromatography was a Kinetex C18 50×2.1 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection and positive electrospray ionization. The MS range was 100 - 1000.)

[0461] General procedure for the preparation of Series A5 - 3C (NUCC - 0226603) - ET43318 - 197

[0462]

Chemical Structure

[0463] Method of prep-HPLC: Equipment: Gilson 281 semi-preparative HPLC system Mobile phase: A: 10 mM NH4HCO3 in H2O; B: acetonitrile Column: Waters Xbridge BEH C18 100×30 mm×10 μm Flow rate: 25 mL / min Monitoring wavelength: 220 and 254 nm

Table 6

[0464] 5_95CD_6 min - 220 - 254 - ELSD 5_95CD_6 min - 220 - 254 - ELSD: LC / MS (The gradient was 5% B from 0.40 min, 5 - 95% B from 0.40 to 3.40 min, held at 95% B for 0.45 min, then 95 - 5% B in 0.01 min, and the flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3 and mobile phase B was acetonitrile. The column used for chromatography was an Xbridge Shield RP18 2.1×50 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection as well as positive electrospray ionization. The MS range was 100 - 1000.)

[0465] Synthesis of A4 - 3 (NUCC - 0226596) [Chemical Structure] Synthesis Scheme [Chemical Structure] Chemical Synthesis LCMS Method: Method 1: 5_95CD_6 min - 220 - 254 - ELSD LC / MS (The gradient was 5% B from 0.40 min, 5 - 95% B from 0.40 to 3.40 min, held at 95% B for 0.45 min, then 95 - 5% B in 0.01 min, and the flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3 and mobile phase B was acetonitrile. The column used for chromatography was an Xbridge Shield RP18 2.1×50 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection as well as positive electrospray ionization. The MS range was 100 - 1000.)

[0466] Experiment for the Largest - Scale Execution: General Procedure for the Preparation of Series A4 - 3 (NUCC - 0226596) - ET43318 - 189

[0467] [Chemical Structure] To a solution of D-7 (25 mg, 28.70 μmol) and 2,6-dimethylpyridine (61.50 mg, 0.574 mmol) in DCM (0.5 mL), TMSBr (43.93 mg, 0.287 mmol) was added dropwise at 0 °C. The mixture was warmed to 20 °C and stirred for 12 hours. LCMS indicated that the starting material was consumed and 18.3% of the product with the desired Ms was detected. The mixture was filtered to obtain a filtrate. The filtrate was purified by prep-HPLC to give Series A4-3 (3.4 mg, yield 15.36%) as a white solid.

[0468] Method for prep-HPLC: Equipment: Gilson 281 semi-preparative HPLC system Mobile phase: A: 10 mM NH4HCO3 in H2O; B: acetonitrile Column: Waters Xbridge BEH C18 100×30 mm×10 μm Flow rate: 25 mL / min Monitoring wavelength: 220 and 254 nm

Table 7

[0469] 5_95CD_6 min - 220 - 254 - ELSD 5_95CD_6 min - 220 - 254 - ELSD: LC / MS (The gradient was 5% B at 0.40 min, 5 - 95% B from 0.40 to 3.40 min, held at 95% B for 0.45 min, then 95 - 5% B in 0.01 min, and the flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3 and mobile phase B was acetonitrile. The column used for chromatography was an Xbridge Shield RP18 2.1×50 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection as well as positive electrospray ionization. The MS range was 100 - 1000.)

[0470] Synthesis of A4 - 3 (NUCC - 0226596) [Chemical formula] Synthesis scheme [Chemical formula] Chemical synthesis LCMS method: Method 1: 5_95CD_6 min - 220 - 254 - ELSD LC / MS (The gradient was 5% B at 0.40 min, 5 - 95% B from 0.40 to 3.40 min, held at 95% B for 0.45 min, then 95 - 5% B in 0.01 min, and the flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3 and mobile phase B was acetonitrile. The column used for chromatography was an Xbridge Shield RP18 2.1×50 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection as well as positive electrospray ionization. The MS range was 100 - 1000.)

[0471] Experiment for the largest - scale execution: General procedure for the preparation of series A4 - 3 - ET43318 - 196

[0472] [Chemical formula] To a solution of D-7 (60 mg, 68.88 μmol) and 2,6-dimethylpyridine (295.21 mg, 2.76 mmol) in DCM (2 mL), TMSBr (210.89 mg, 1.38 mmol) was added dropwise at 0 °C. The mixture was warmed to 20 °C and stirred for 2 hours. LCMS indicated that the starting material was consumed and 18.6% of the product with the desired Ms was detected. The mixture was filtered to obtain a filtrate. The filtrate was purified by prep-HPLC to give Series A4-3 (11.2 mg, yield 21.43%) as a white solid.

[0473] Method of prep-HPLC: Equipment: Gilson 281 semi-preparative HPLC system Mobile phase: A: 10 mM NH4HCO3 in H2O; B: acetonitrile Column: Waters Xbridge BEH C18 100×30 mm×10 μm Flow rate: 25 mL / min Monitoring wavelength: 220 and 254 nm

Table 8

[0474] 5_95CD_6 min - 220 - 254 - ELSD 5_95CD_6 min - 220 - 254 - ELSD: LC / MS (Gradient: 5% B from 0.40 min, 5 - 95% B from 0.40 - 3.40 min, hold at 95% B for 0.45 min, then 95 - 5% B in 0.01 min, flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3 and mobile phase B was acetonitrile. The column used for chromatography was an Xbridge Shield RP18 2.1×50 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection and positive electrospray ionization. The MS range was 100 - 1000.)

[0475] Synthesis of D - 7 (NUCC - 0226597)

Chemical Structure

Chemical Structure

[0476] Experiment for the Largest - Scale Execution: General Procedure for the Preparation of Compound D - 3 - ET43318 - 118

[0477] [Chemistry] To a solution of 1-[diethoxyphosphorylmethyl(ethoxy)phosphoryl]oxyethane (10 g, 34.70 mmol) in THF (100 mL), NaH (1.53 g, 38.17 mmol) was added portionwise at 0 °C under nitrogen. After stirring for 30 minutes, D-2 (9.29 g, 41.64 mmol) was added. The mixture was heated to 70 °C and stirred for 12 hours. TLC (ethyl acetate / MeOH = 5 / 1, Rf = 0.55) indicated that the starting material was consumed and a new spot appeared. The reaction mixture was poured into NH4Cl (300 mL) and extracted with ethyl acetate (3 × 100 mL). The combined organic layers were washed with brine (2 × 50 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give D-3 (12 g, crude) as a yellow oil, which was used directly in the next step without further purification.

[0478] 1 H NMR (400 MHz, chloroform-d) δ 1.32 - 1.37 (m, 12H), 1.55 - 1.61 (m, 4H), 1.73 (br dd, J = 6.50, 3.00 Hz, 1H), 1.81 - 1.92 (m, 3H), 1.96 - 2.09 (m, 2H), 2.26 - 2.48 (m, 1H), 3.52 - 3.55 (m, 2H), 3.85 - 3.90 (m, 2H), 4.15 - 4.23 (m, 8H), 4.58 - 4.61 (m, 1H)

[0479] General procedure for the preparation of compound D-4-ET43318-137 [Chemistry] To a solution of D-3 (5 g, 11.62 mmol) in MeOH (50 mL) was added 4-methylbenzenesulfonic acid (100.02 mg, 580.84 μmol) at 20 °C. The mixture was stirred for 12 h. TLC (ethyl acetate / MeOH = 5:1, Rf = 0.4) indicated that the starting material was consumed and a new spot had formed. The mixture was concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (eluting with ethyl acetate / methanol = 20 / 1 to 5 / 1) to afford D-4 (1.2 g, yield 26.85%) as a colorless oil.

[0480] 1 H NMR (400 MHz, chloroform-d) δ 1.35 (t, J = 7.00 Hz, 12H), 1.75 - 1.89 (m, 2H), 2.00 - 2.13 (m, 2H), 2.38 - 2.47 (m, 1H), 3.67 (t, J = 5.94 Hz, 2H), 4.19 (dq, J = 13.80, 7.03 Hz, 8H)

[0481] General procedure for the preparation of compound D-5-ET43318-152

Chemical formula

[0482] 1 H NMR (400 MHz, chloroform-d) δ 1.34 (td, J = 7.04, 1.44 Hz, 12H), 2.07 - 2.20 (m, 4H), 2.29 - 2.45 (m, 1H), 3.88 (s, 3H), 4.03 (t, J = 5.50 Hz, 2H), 4.15 - 4.23 (m, 8H), 6.84 - 6.94 (m, 2H), 7.93 - 8.03 (m, 2H)

[0483] General procedure for the preparation of compound Linker D3-ET43318-158

Chemical Structure

[0484] 1 H NMR (400 MHz, chloroform-d) δ 1.34 (td, J = 7.07, 1.50 Hz, 12H), 2.06 - 2.21 (m, 4H), 2.30 - 2.46 (m, 1H), 3.98 (t, J = 5.69 Hz, 2H), 4.14 - 4.24 (m, 8H), 4.62 (s, 2H), 6.84 - 6.90 (m, 2H), 7.27 - 7.31 (m, 2H)

[0485] General procedure for the preparation of compound D-6-ET43318-163

Chemical Structure

[0486] 1 H NMR (400 MHz, chloroform-d) δ 0.94 (s, 12H), 1.77 (s, 3H), 2.06 - 2.18 (m, 4H), 2.29 - 2.46 (m, 1H), 3.79 - 3.86 (m, 3H), 3.97 (br t, J = 5.44 Hz, 2H), 4.14 - 4.24 (m, 8H), 4.99 - 5.08 (m, 2H), 6.45 - 6.57 (m, 1H), 6.80 - 6.88 (m, 2H), 7.08 (d, J = 8.63 Hz, 1H), 7.12 - 7.19 (m, 2H), 7.32 (d, J = 8.63 Hz, 1H), 7.42 - 7.48 (m, 1H), 7.49 - 7.56 (m, 1H), 7.71 (d, J = 1.38 Hz, 1H)

[0487] General procedure for the preparation of compound D-7 (NUCC-0226597)-ET43318-169

Chemical Structure

[0488] Method for prep-HPLC: Equipment: Gilson 281 semi-preparative HPLC system Mobile phase: A: 10 mM NH4HCO3 in H2O; B: acetonitrile Column: Waters Xbridge BEH C18 100×30 mm×10 μm Flow rate: 25 mL / min Monitoring wavelength: 220 and 254 nm

Table 9

[0489] 5_95AB_6 min - 220 - 254 - ELSD 5_95AB_6 min - 220 - 254 - ELSD: LC / MS (The gradient was 5% B for 0.40 min, 5 - 95% B from 0.40 to 3.00 min, held at 95% B for 1.00 min, then 95 - 5% B in 0.01 min, and the flow rate was 1.0 mL / min. Mobile phase A was 0.037% trifluoroacetic acid in water, and mobile phase B was 0.018% trifluoroacetic acid in acetonitrile. The column used for chromatography was a Kinetex C18 50×2.1 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection and positive electrospray ionization. The MS range was 100 - 1000.)

[0490] Synthesis of NUCC - 0226914 (ET43587 - 416 - 1) [Chemical Structure Diagram] Chemical synthesis LCMS method: Method 1: 5_95AB_2 min LC / MS (The column used for chromatography was a Kinetex EVO C18 2.1×30 mm, 5 μm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in HPLC - grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B (0.70 - 1.16 min), 95 - 5% B (1.16 - 1.50 min). The flow rate was 1.5 mL / min.)

[0491] Method 2: 5_95AB_6 min - 220 - 254 - ELSD LC / MS (Gradient: 5% B in 0.40 min, 5 - 95% B in 2.60 min, hold at 95% B for 1.00 min, then 95 - 5% B in 0.01 min, flow rate 1.0 mL / min. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in acetonitrile. The column used for chromatography was Kinetex C18 2.1×50 mm, 5 μm. The detection methods were diode array (DAD) and evaporative light scattering detection (ELSD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0492] Experiment for the largest scale execution: General procedure for the preparation of compound 15 - ET43587 - 401

[0493] [Chemical formula] To a solution of compound 14 (200 mg, 299.59 μmol, 1 equiv) in DMF (4 mL), NH4Cl (80.13 mg, 1.50 mmol, 5 equiv) and DIEA (116.16 mg, 898.78 μmol, 156.55 μL, 3 equiv) were added HOBt (60.72 mg, 449.39 μmol, 1.5 equiv), and the mixture was stirred at 25 °C for 30 min. Then, EDCI (86.15 mg, 449.39 μmol, 1.5 equiv) was added at 0 °C. The mixture was stirred at 25 °C for 12 h. LCMS indicated that all starting materials were consumed and a new peak with the desired product MS was detected. The reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (3×10 mL). The organic layers were separated, combined organic layers were washed with brine (2×10 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain a residue, which was purified by silica gel chromatography (eluting with petroleum ether / ethyl acetate = 20 / 1 - 1 / 3) to give compound 15 (170 mg, yield 76.61%) as a white solid.

[0494] 11H NMR (ET43587-401-P1A, 400 MHz, chloroform-d) δ 0.10 (s, 9H), 0.51 - 0.59 (m, 2H), 2.85 - 2.96 (m, 2H), 3.85 (s, 3H), 4.41 (s, 2H), 5.06 (s, 2H), 5.34 (br s, 1H), 6.75 (br s, 1H), 6.85 (s, 1H), 6.91 (d, J = 8.63 Hz, 1H), 7.17 (d, J = 8.50 Hz, 2H), 7.28 - 7.35 (m, 3H), 7.53 - 7.61 (m, 2H), 7.84 (d, J = 1.63 Hz, 1H)

[0495] General procedure for the preparation of compound 16 - ET43587-406 [Chemical formula] A mixture of compound 15 (100 mg, 150.02 μmol, 1 equivalent) in TBAF (1 M, 3.00 mL, 20 equivalents) was stirred at 25 °C for 20 h. LCMS indicated that all starting materials were consumed and a new peak with the desired product MS was detected. The reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (3 × 10 mL). The organic layers were separated, combined organic layers were washed with brine (10 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue, which was purified by chromatography on silica gel (eluting with petroleum ether / ethyl acetate = 20 / 1 - 1 / 2) to give compound 16 (50 mg, yield 55.93%) as a yellow solid.

[0496] LCMS (ESI+): RT = 0.846 min, m / z 536.1 (M + H) +

[0497] 5_95 AB_2 minutes: For LC / MS, the column used in chromatography was Kinetex EVO C18 2.1×30 mm, 5 μm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in HPLC grade acetonitrile. The gradient was 5 - 95% B in 1.50 minutes, 5% B in 0.01 minute, 5 - 95% B (0.01 - 0.70 minutes), 95% B (0.70 - 1.16 minutes), 95 - 5% B (1.16 - 1.50 minutes). The flow rate was 1.5 mL / min.

[0498] General procedure for the preparation of Series 11 - 3 - 12 - ET43587 - 416

[0499]

Chemical Structure

[0500] 11H NMR (ET43587-416-P1A, 400 MHz, chloroform-d) δ 3.85 (s, 3H), 5.06 (s, 2H), 6.70 (s, 1H), 6.73 (d, J = 8.63 Hz, 1H), 7.16 (d, J = 8.38 Hz, 2H), 7.21 (d, J = 8.63 Hz, 1H), 7.29 - 7.34 (m, 2H), 7.53 (dd, J = 8.13, 2.00 Hz, 1H), 7.65 (d, J = 8.25 Hz, 1H), 7.79 (d, J = 1.88 Hz, 1H) LCMS (ESI+): RT = 3.252 min, m / z 518.0 (M + H) +

[0501] 5_95AB_6 min - 220 - 254 - ELSD: LC / MS (Gradient was 5% B in 0.40 min, 5 - 95% B in 2.60 min, hold 95% B for 1.00 min, then 95 - 5% B in 0.01 min, and the flow rate was 1.0 mL / min. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in acetonitrile. The column used for chromatography was Kinetex C18 2.1×50 mm, 5 μm. The detection methods were diode array (DAD) and evaporative light scattering detection (ELSD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0502] Synthesis of NUCC - 0226659 (ET43587 - 299 - 1) and NUCC - 0226660 (ET43587 - 309 - 1)

Chemical Structure

Chemical Structure

[0503] The inventors take Series 11 - 3 - 9 (NUCC - 0226659) as an example.

[0504] Experiment for the largest - scale execution: General procedure for the preparation of compound 6 - ET43587 - 275

[0505] The reactions were carried out in parallel and combined for purification.

[0506] [Chemical Structure] To a solution of Series 11 - 3 - core 1 (100 mg, 203.21 μmol, 1 equiv) and DIEA (78.79 mg, 609.62 μmol, 106.18 μL, 3 equiv) in DCM (1 mL) was added SEMCl (67.76 mg, 406.41 μmol, 71.93 μL, 2 equiv) at 0 °C. The reaction mixture was stirred at 25 °C for 12 h. TLC (petroleum ether / ethyl acetate = 10 / 1) showed that the starting material (R f = 0.45) was consumed and a new spot (R f = 0.5) was formed. Three additional reactions were set up as described above and all four reaction mixtures were combined. The combined reaction mixture was concentrated under reduced pressure to give a residue, which was purified by preparative TLC (petroleum ether / ethyl acetate = 10 / 1) to give compound 6 (340 mg, yield 60.49%) as a white solid.

[0507] 1 1H NMR (ET43587 - 275 - P2A, 400 MHz, chloroform - d) δ - 0.045 (s, 9H), 0.62 - 0.71 (m, 2H), 3.20 - 3.28 (m, 2H), 4.83 (s, 2H), 4.98 (s, 2H), 6.73 (d, J = 8.88 Hz, 1H), 7.12 (d, J = 8.50 Hz, 2H), 7.28 - 7.31 (m, 2H), 7.51 - 7.56 (m, 3H), 7.81 (s, 1H)

[0508] General procedure for the preparation of compound 7 - ET43587 - 288 [Chemical formula] To a solution of compound 6 (70 mg, 112.47 μmol, 1 equiv) and compound 11 - 3 - 9A (167.42 mg, 449.89 μmol, 4 equiv) in dioxane (4 mL), Pd(dppf)Cl2 (16.46 mg, 22.49 μmol, 0.2 equiv) was added under nitrogen, and the mixture was stirred at 100 °C for 12 h. TLC (petroleum ether / ethyl acetate = 1 / 1) indicated that the starting material (R f = 0.9) was consumed and a new spot (R f = 0.3) was formed. The reaction mixture was filtered and concentrated under reduced pressure to give a residue, which was purified by preparative TLC (petroleum ether / ethyl acetate = 1 / 1) to afford compound 7 (120 mg, yield 76.87%) as a white solid.

[0509] 1 1H NMR (ET43587 - 288 - P1A, 400 MHz, methanol - d4) δ - 0.1 (s, 9H), 0.49 - 0.57 (m, 2H), 2.87 - 2.96 (m, 2H), 4.04 (s, 3H), 4.38 (s, 2H), 5.14 (s, 2H), 7.16 (d, J = 8.63 Hz, 1H), 7.23 - 7.34 (m, 4H), 7.42 (d, J = 8.63 Hz, 1H), 7.64 - 7.72 (m, 2H), 7.76 - 7.81 (m, 1H), 7.86 (s, 1H)

[0510] General Procedure for the Preparation of Series 11-3-9-ET43587-299

Chem.

[0511] 1 H NMR (ET43587-299-P1A, 400 MHz, methanol-d4) δ 3.97 (s, 3H), 5.10 (s, 2H), 6.87 (d, J = 8.63 Hz, 1H), 7.19 - 7.32 (m, 5H), 7.58 - 7.80 (m, 4H)

[0512] LCMS (ESI+): RT = 3.516 min, m / z 494.1 (M + H) +

[0513] 5_95CD_6 min - 220 - 254 - ELSD: LC / MS (gradient was 5% B at 0.40 min, 5 - 95% B from 0.40 - 3.40 min, 95% B held for 0.45 min, then 95 - 5% B in 0.01 min, and the flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3 and mobile phase B was acetonitrile. The column used for chromatography was an Xbridge C18 2.1×50 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0514] Synthesis of NUCC-0226656 (ET43587-277-1) and NUCC-0226657 (ET43587-268-1)

Chemical formula

[0515] Method 2: 5_95CD_6 minutes - 220 - 254 - ELSD LC / MS (The gradient was 5%B in 0.40 minutes, 5 - 95%B from 0.40 to 3.40 minutes, held at 95%B for 0.45 minutes, then 95 - 5%B in 0.01 minute, and the flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3, and mobile phase B was acetonitrile. The column used for chromatography was an Xbridge C18 2.1×50mm column (5μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0516] Method 3: 5_95AB_6 minutes - 220 - 254 - ELSD LC / MS (Gradient: 5% B in 0.40 min, 5 - 95% B in 2.60 min, hold at 95% B for 1.00 min, then 95 - 5% B in 0.01 min; flow rate: 1.0 mL / min. Mobile phase A: 0.04% trifluoroacetic acid in water; mobile phase B: 0.02% trifluoroacetic acid in acetonitrile. Column used for chromatography: Kinetex C18 2.1×50 mm, 5 μm. Detection methods: diode array (DAD), evaporative light scattering detection (ELSD). MS mode: positive electrospray ionization. MS range: 100 - 1000.)

[0517] Experiment for the largest scale execution: General procedure for the preparation of Series 11 - 3 - Core 1 - ET42365 - 239

[0518]

Chemical formula

[0519] LCMS (ESI+): RT = 1.042 min, m / z 490.9 (M - H) -

[0520] NEG5-95CD_2 fractions: LC / MS (The column used in chromatography was Xbridge C18 2.1×50mm, 5μm. The detection method was diode array (DAD). The MS mode was negative electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 10 mM ammonium bicarbonate in water, and mobile phase B was HPLC grade acetonitrile. The gradient was 5 - 95%B in 1.50 minutes, 5%B in 0.01 minutes, 5 - 95%B (0.01 - 0.70 minutes), 95%B (0.70 - 1.16 minutes), 95 - 5%B (1.16 - 1.50 minutes), held at 5%B for 0.34 minutes, and the flow rate was 1.5 mL / min.)

[0521] General procedure for the preparation of Series 11-3-13-ET43587-277

[0522] [Chemical formula] To a solution of Series 11-3-Core 1 (150 mg, 304.81 μmol, 1 equivalent), 11-3-13A (164.89 mg, 792.50 μmol, 2.6 equivalents), and K3PO4 (194.10 mg, 914.42 μmol, 3 equivalents) in a mixed solution of THF (5 mL) and H2O (1 mL), di-tert-butyl(cyclopentyl)phosphane; dichloropalladium; iron (39.73 mg, 60.96 μmol, 0.2 equivalent) was added under nitrogen, and the reaction mixture was stirred at 80 °C for 12 hours. LCMS indicated that the starting materials were consumed and a product with 67.7% of the desired MS was detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain a residue, which was purified by prep-HPLC (column: Waters Xbridge Prep OBD C18 150×40mm×10μm; mobile phase: [water (NH4HCO3)-ACN]; B%: 70% - 95%, 8 minutes) to obtain Series 11-3-13 (11.5 mg, yield 7.28%) as a white solid.

[0523] 11H NMR (ET43587-277-P1A, 400 MHz, methanol-d4) 3.73 (s, 3H), 5.08 (s, 2H), 6.29 (d, J = 1.88 Hz, 1H), 6.83 (d, J = 8.63 Hz, 1H), 7.19 - 7.23 (m, 3H), 7.27 - 7.31 (m, 2H), 7.49 (d, J = 2.00 Hz, 1H), 7.58 - 7.65 (m, 2H), 7.77 (s, 1H) LCMS (ESI+): RT = 3.650 min, m / z 493.1 (M + H) +

[0524] 5_95 CD_6 min - 220 - 254 - ELSD: LC / MS (Gradient was 5% B in 0.40 min, 5 - 95% B from 0.40 - 3.40 min, held at 95% B for 0.45 min, then 95 - 5% B in 0.01 min, flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3 and mobile phase B was acetonitrile. The column used for chromatography was an Xbridge C18 2.1×50 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0525] General procedure for the preparation of Series 11 - 3 - 16 - ET43587 - 268

[0526]

Chemical Structure

[0527] 1 H NMR (ET43587-268-P1, 400 MHz, methanol-d4) δ 1.42 (d, J = 6.63 Hz, 6H), 4.44 (quin, J = 6.63 Hz, 1H), 5.09 (s, 2H), 6.51 (s, 1H), 6.84 (d, J = 8.50 Hz, 1H), 7.19 - 7.24 (m, 3H), 7.27 - 7.32 (m, 2H), 7.57 - 7.66 (m, 2H), 7.77 (d, J = 1.75 Hz, 1H) LCMS (ESI+): RT = 3.475 min, m / z 589.0 (M + H) +

[0528] 5_95AB_6 min - 220 - 254 - ELSD: LC / MS (Gradient: 5% B at 0.40 min, 5 - 95% B at 2.60 min, hold at 95% B for 1.00 min, then 95 - 5% B in 0.01 min, flow rate was 1.0 mL / min. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in acetonitrile. The column used for chromatography was Kinetex C18 2.1×50 mm, 5 μm. The detection methods were diode array (DAD) and evaporative light scattering detection (ELSD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0529] Synthesis of NUCC - 0226652 (ET42365 - 279 - 1) [Chemical formula] Synthesis scheme: Preparation of Series 11 - 4 - 2 (NUCC - 0226652) [Chemical formula] Chemical synthesis NEG5 - 95CD_2 min: LC / MS (The column used for chromatography was Xbridge C18 2.1×50 mm, 5 μm. The detection method was diode array (DAD). The MS mode was negative electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 10 mM ammonium bicarbonate in water, and mobile phase B was HPLC - grade acetonitrile. The gradient was 5 - 95% B at 1.50 min, 5% B at 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B (0.70 - 1.16 min), 95 - 5% B (1.16 - 1.50 min), hold at 5% B for 0.34 min, and the flow rate was 1.5 mL / min.)

[0530] 50_100AB_6 min - 220 - 254 - ELSD: LC / MS (Gradient was 50% B at 0.40 min, 50 - 100% B from 0.40 to 3.00 min, held at 100% B for 1.00 min, then 100 - 50% B in 0.01 min, flow rate was 1.0 mL / min. Mobile phase A was 0.04% trifluoroacetic acid in water and mobile phase B was 0.02% trifluoroacetic acid in acetonitrile. The column used for chromatography was a Luna C18 50×2.0 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection as well as positive electrospray ionization. The MS range was 100 - 1000.)

[0531] Experiment for the largest scale execution: General procedure for the preparation of compound 2 - ET43365 - 250

[0532] [Chemical formula] To a solution of compound 1 (0.5 g, 2.89 mmol, 335.57 μL, 1 equivalent), compound 2A (587.51 mg, 3.76 mmol, 1.3 equivalents) and K3PO4 (1.53 g, 7.23 mmol, 2.5 equivalents) in a mixed solution of THF (10 mL) and H2O (2 mL), Pd(dppf)Cl2·CH2Cl2 (177.01 mg, 216.75 μmol, 0.075 equivalent) was added under nitrogen and the reaction mixture was stirred at 80 °C for 2 h. LCMS indicated that the starting materials were consumed and a new peak with the desired product Ms was detected. The crude product was diluted with water (50 mL) and extracted with ethyl acetate (3×50 mL). The organic layer was washed with brine (50 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography on silica gel (eluting with petroleum ether / ethyl acetate = 50 / 1 - 10 / 1) to give compound 2 (500 mg, yield 80.31%) as a yellow solid.

[0533] 11H NMR (ET42365-250-P1A, 400 MHz, chloroform-d) δ 4.96 (s, 1H), 6.85 - 6.96 (m, 1H), 7.12 - 7.22 (m, 1H), 7.31 - 7.42 (m, 2H) General procedure for the preparation of compound 3 - ET42365-256

[0534] [Chemical formula] To a solution of compound 2 (500 mg, 2.44 mmol, 1 equiv) and DIPA (494.45 mg, 4.89 mmol, 690.57 μL, 2 equiv) in CHCl3 (12.5 mL) was added 1-bromopyrrolidine-2,5-dione (413.10 mg, 2.32 mmol, 0.95 equiv) at -40 °C, and the reaction mixture was stirred at -40 °C for 4 h. LCMS indicated that approximately 9.6% of the starting material remained and a new peak (61.8%) was detected. The reaction mixture was diluted with water (50 mL) and extracted with DCM (3 × 30 mL). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, and concentrated to give the crude product. The crude product was purified by column chromatography on silica gel (eluting with petroleum ether / ethyl acetate = 50 / 1 to 10 / 1) to afford compound 3 (500 mg, yield 64.96%) as a yellow solid.

[0535] 1 1H NMR (ET42365-256-P1A, 400 MHz, chloroform-d) δ 5.68 (s, 1H), 6.90 (t, J = 7.84 Hz, 1H), 7.24 (dd, J = 7.65, 1.38 Hz, 1H), 7.39 - 7.45 (m, 2H), 7.46 - 7.52 (m, 3H)

[0536] General procedure for the preparation of compound 4 - ET42365-257 [Chemical formula] To a solution of Compound 3 (500 mg, 1.76 mmol, 1 equiv) in DMF (12 mL) was added K2CO3 (487.42 mg, 3.53 mmol, 2 equiv) and Compound 3A (530.47 mg, 1.94 mmol, 1.1 equiv), and the reaction mixture was stirred at 25 °C for 12 h. LCMS indicated that the starting material was consumed and a new peak was detected. The crude product was diluted with water (50 mL) and extracted with ethyl acetate (3 × 30 mL). The organic layer was washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford Compound 4 (700 mg, yield 75.04%) as a yellow oil. The product was used directly in the next step without further purification.

[0537] 1 H NMR (ET42365-257-P1A, 400 MHz, chloroform-d) δ 4.59 (s, 2H), 7.08 - 7.15 (m, 1H), 7.25 (dd, J = 8.22, 1.82 Hz, 1H), 7.27 - 7.30 (m, 1H), 7.35 - 7.48 (m, 6H), 7.60 (dd, J = 7.97, 1.57 Hz, 1H)

[0538] General procedure for the preparation of Compound 5-ET42365-260 The reactions were carried out in parallel but combined for purification.

[0539]

Chemical Structure

[0540] 1 H NMR (ET42365 - 260 - P1A, 400 MHz, chloroform - d) δ 4.57 (s, 2H), 5.66 (s, 1H), 6.91 (d, J = 7.63 Hz, 1H), 7.02 (d, J = 8.00 Hz, 1H), 7.12 - 7.19 (m, 1H), 7.26 (br d, J = 8.13 Hz, 1H), 7.41 - 7.50 (m, 4H), 7.54 (d, J = 8.38 Hz, 2H)

[0541] General procedure for the preparation of compound 6 - ET42365 - 274

Chemical Structure

[0542] LCMS (ESI+): RT = 1.061 min, m / z 490.8 (M-H) +

[0543] NEG5-95CD_2 min: The column used for chromatography was Xbridge C18 2.1×50 mm, 5 μm. The detection method was diode array (DAD). The MS mode was negative electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 10 mM ammonium bicarbonate in water and mobile phase B was HPLC grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B for 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B (0.70 - 1.16 min), 95 - 5% B (1.16 - 1.50 min), held at 5% B for 0.34 min, and the flow rate was 1.5 mL / min.

[0544] General procedure for the preparation of Series 11-4-2-ET42365-279

[0545]

Chemical Structure

[0546] 1 H NMR (ET42365-279-P1A, 400 MHz, chloroform-d) δ 3.82 (s, 3H), 4.59 (s, 2H), 6.02 (s, 1H), 6.58 (s, 1H), 6.99 (d, J = 8.03 Hz, 1H), 7.11 (d, J = 8.16 Hz, 1H), 7.25 (s, 1H), 7.41 - 7.51 (m, 4H), 7.54 - 7.61 (m, 2H) LCMS (ESI+): RT = 2.774 min, m / z 561.0 (M + H) +

[0547] 50_100AB_6 min - 220 - 254 - ELSD: LC / MS (Gradient: 50% B at 0.40 min, 50 - 100% B from 0.40 to 3.00 min, hold at 100% B for 1.00 min, then 100 - 50% B in 0.01 min, flow rate was 1.0 mL / min. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in acetonitrile. The column used for chromatography was Luna C18 50×2.0 mm column (5 μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection and positive electrospray ionization. The MS range was 100 - 1000.)

[0548] Synthesis of NUCC - 0226606 (ET42365 - 228 - P1) and NUCC - 0226605 (ET42365 - 228 - P2)

Chem.

Chem.

[0549] Method 2: 5 - 95 CD_4.5 min: LC / MS (The column used in chromatography was Xbridge C18 2.1×50 mm, 5 μm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 10 mM ammonium bicarbonate in water, and mobile phase B was HPLC - grade acetonitrile. The gradient was 5 - 95% B at 4.30 min, 5% B at 0.01 min, 5 - 95% B (0.01 - 3.00 min), held at 95% B within 0.5 min, 95 - 5% B (3.50 - 3.51 min), and held at 5% B for 0.79 min. The flow rate was 1.0 mL / min (0.01 - 4.30 min).

[0550] Method 3: 50_100 CD_6 min - 220 - 254 - ELSD: LC / MS (The gradient was 50% B at 0.40 min, 50 - 100% B from 0.40 to 3.40 min, held at 100% B for 0.45 min, then 100 - 50% B at 0.01 min, and the flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3, and mobile phase B was acetonitrile. The column used in chromatography was an Xbridge C18 2.1×50 mm column (5 - μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0551] Method 4: 5_95 CD_6 min - 220 - 254 - ELSD: LC / MS (The gradient was 5% B at 0.40 min, 5 - 95% B from 0.40 to 3.40 min, held at 95% B for 0.45 min, then 95 - 5% B at 0.01 min, and the flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3, and mobile phase B was acetonitrile. The column used in chromatography was an Xbridge C18 2.1×50 mm column (5 - μm particles). The detection method was diode array (DAD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0552] Experiment for maximum scale execution: General procedure for the preparation of compound 13 - ET42365 - 161

[0553] [Chemical formula] To a solution of compound 11 (300 mg, 2.33 mmol, 1.2 equiv) in THF (6 mL) was added t - BuOK (1 M, 2.33 mL, 1.2 equiv) at 0 °C. The reaction mixture was stirred at 0 °C for 1 h, then compound 12 (390.56 mg, 1.94 mmol, 1 equiv) was added, and the reaction mixture was stirred at 50 °C for 12 h. LCMS indicated that all starting materials were consumed and two new peaks were detected. The reaction mixture was filtered, and the solid was collected and dried to give compound 13 (300 mg, yield 62.08%) as an off - white solid.

[0554] 1 H NMR (ET42365 - 161 - P1B, 400 MHz, DMSO - d6) δ 2.94 (q, J = 5.09 Hz, 2H), 6.82 - 6.89 (m, 2H), 7.17 - 7.24 (m, 2H)

[0555] General procedure for the preparation of compound 7 - ET42365 - 170 [Chemical formula] To a solution of core A3_1 (400 mg, 835.47 μmol, 1 equiv) and pyridine (264.34 mg, 3.34 mmol, 269.74 μL, 4 equiv) in DCM (12 mL) was added dropwise Tf2O (353.58 mg, 1.25 mmol, 206.77 μL, 1.5 equiv) at 0 °C, and the reaction mixture was stirred at 20 °C for 3 h. LCMS indicated that the starting material was consumed and a new peak with the desired product Ms was detected. The reaction mixture was diluted with water (50 mL) and extracted with DCM (3 × 25 mL). The organic layers were separated, combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the crude product. The crude product was purified by column chromatography on silica gel (eluting with petroleum ether / ethyl acetate = 50 / 1 - 10 / 1) to give compound 7 (0.4 g, yield 78.38%) as a white solid.

[0556] LCMS (ESI+): RT = 0.915 min, m / z 611.1 (M+H) +

[0557] 5 - 95 AB_2 min: LC / MS (The column used for chromatography was an Agilent Poroshell SB-C18 3.0×30 mm, 2.7 μm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water and mobile phase B was 0.02% trifluoroacetic acid in HPLC grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B (0.70 - 1.16 min), 95 - 5% B (1.16 - 1.50 min). The flow rate was 1.5 mL / min.

[0558] General procedure for the preparation of compound 8 - ET42365 - 200

[0559]

Chemical Structure

[0560] LCMS (ESI+): RT = 0.988 min, m / z 603.2 (M+H) +

[0561] 5-95AB_2 min: LC / MS (The column used for chromatography was Agilent Poroshell SB-C18 3.0×30 mm, 2.7 μm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in HPLC grade acetonitrile. The gradient was 5 - 95% B in 1.50 min, 5% B in 0.01 min, 5 - 95% B (0.01 - 0.70 min), 95% B (0.70 - 1.16 min), 95 - 5% B (1.16 - 1.50 min). The flow rate was 1.5 mL / min.

[0562] General procedure for the preparation of Compound 8-ET42365-201

[0563]

Chemical formula

[0564] LCMS (ESI+): RT = 3.108 min, m / z 561.0 (M + H) +

[0565] 5 - 95 CD_4.5 min: The column used for LC / MS was Xbridge C18 2.1 × 50 mm, 5 μm. The detection method was diode array (DAD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000. Mobile phase A was 10 mM ammonium bicarbonate in water, and mobile phase B was HPLC grade acetonitrile. The gradient was 5 - 95% B at 4.30 min, 5% B at 0.01 min, 5 - 95% B (0.01 - 3.00 min), hold at 95% B within 0.5 min, 95 - 5% B (3.50 - 3.51 min), and hold at 5% B for 0.79 min. The flow rate was 1.0 mL / min (0.01 - 4.30 min).

[0566] General procedure for the preparation of A02B01C07D01_P1 and A02B01C07D01 - ET42365 - 228

[0567]

Chemical Structure

[0568] A02B01C07D01(NUCC-0226606): 1 H NMR(ET42365-228-P1B,400MHz, chloroform-d) δ 3.84(s,3H),4.65 - 4.78(m,2H),4.93(br s,1H),6.75(d,J = 8.88Hz,2H),7.23(d,J = 8.88Hz,2H),7.33 - 7.43(m,2H),7.54(br d,J = 8.25Hz,1H),7.67(d,J = 8.13Hz,1H),7.76(d,J = 1.38Hz,1H) LCMS(ESI+):RT = 2.467 min,m / z 596.9(M + H) +

[0569] 5_95CD_6 min - 220 - 254 - ELSD: LC / MS (Gradient: 5% B at 0.40 min, 5 - 95% B from 0.40 to 3.40 min, hold at 95% B for 0.45 min, then 95 - 5% B in 0.01 min, flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3, and mobile phase B was acetonitrile. The column used for chromatography was an Xbridge C18 2.1×50 mm column (5 μm particles). The detection method was diode array (DAD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0570] A02B01C07D01(NUCC - 0226605): 1 H NMR (ET42365 - 228 - P1C, 400 MHz, chloroform - d) δ 3.90 (s, 3H), 4.71 - 4.78 (m, 1H), 4.79 - 4.86 (m, 1H), 5.02 (br s, 1H), 6.65 (s, 1H), 6.77 (d, J = 9.01 Hz, 2H), 7.18 - 7.26 (m, 2H), 7.45 (s, 1H), 7.52 (dd, J = 8.19, 1.81 Hz, 1H), 7.62 (d, J = 8.13 Hz, 1H), 7.76 (d, J = 1.63 Hz, 1H) LCMS (ESI+): RT = 2.513 min, m / z 596.9 (M + H) +

[0571] 5_95CD_6 min - 220 - 254 - ELSD: LC / MS (Gradient: 5% B at 0.40 min, 5 - 95% B from 0.40 to 3.40 min, hold at 95% B for 0.45 min, then 95 - 5% B in 0.01 min, flow rate was 0.8 mL / min. Mobile phase A was H2O + 10 mM NH4HCO3, and mobile phase B was acetonitrile. The column used for chromatography was an Xbridge C18 2.1×50 mm column (5 μm particles). The detection method was diode array (DAD) detection. The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0572] Synthesis of A02B01C07D01_P1 (NUCC-0226606) and A02B01C07D01_P2 (NUCC-0227064) [Chemical formula] [Chemical formula] Chemical synthesis LCMS method: 5_95AB_6 min - 220 - 254 - ELSD LC / MS (Gradient: 5%B in 0.40 min, 5 - 95%B from 0.40 to 3.00 min, hold 95%B for 1.00 min, then 95 - 5%B in 0.01 min, flow rate was 1.0 mL / min. Mobile phase A was 0.037% trifluoroacetic acid in water, and mobile phase B was 0.018% trifluoroacetic acid in acetonitrile. The column used for chromatography was a Kinetex C18 50×2.1 mm column (5μm particles). The detection methods were diode array (DAD) and evaporative light scattering (ELSD) detection and positive electrospray ionization. The MS range was 100 - 1000.)

[0573] Experiment for the largest - scale execution: General procedure for the preparation of compound 2 - ET52382 - 5

[0574] [Chemical formula] To a solution of compound 1 (25 g, 108.20 mmol) in MeOH (250 mL) was added sulfuric acid (9.20 g, 93.80 mmol, 5 mL) dropwise at 20 °C. The mixture was heated to 70 °C and stirred for 12 h. TLC (petroleum ether / ethyl acetate = 3 / 1, Rf = 0.25) indicated that the starting material was consumed and a new spot had formed. Two further reactions were set up as above and the three reaction mixtures were combined. The reaction mixture was concentrated to give the crude product, which was diluted with water (500 mL), adjusted to pH = 6 - 7 with K2CO3, and extracted with ethyl acetate (3 × 200 mL). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, and concentrated to give compound 2 (76 g, yield 90.76%) as a white solid, which was used directly in the next step without further purification.

[0575] 1 1H NMR (400 MHz, chloroform-d) δ 3.93 (s, 3H), 3.94 (s, 3H), 7.02 (dd, J = 8.00, 1.38 Hz, 1H), 7.26 - 7.28 (m, 1H), 7.30 - 7.36 (m, 1H)

[0576] General procedure for the preparation of compound 3-ET52382-10

Chemical formula

[0577] 1 1H NMR (400 MHz, chloroform-d) δ 3.62 (s, 3H), 3.77 (s, 3H), 7.14 (d, J = 8.00 Hz, 1H), 7.36 (dd, J = 8.19, 1.44 Hz, 1H), 7.41 - 7.47 (m, 1H), 7.48 - 7.54 (m, 2H), 7.57 (d, J = 1.50 Hz, 1H)

[0578] General procedure for the preparation of compound 4-ET52382-14

Chemical formula

[0579] 1 1H NMR (400 MHz, DMSO-d6) δ 7.12 (d, J = 7.75 Hz, 1H), 7.25 - 7.35 (m, 2H), 7.51 (br d, J = 8.25 Hz, 1H), 7.60 (s, 1H), 7.70 (d, J = 8.25 Hz, 1H), 9.88 (s, 1H), 12.70 (br s, 1H)

[0580] General procedure for the preparation of Compound 5-ET52382-16

Chemical formula

[0581] 1 1H NMR (400 MHz, DMSO-d6) δ 4.15 (d, J = 5.25 Hz, 2H), 5.00 (t, J = 5.32 Hz, 1H), 6.86 (d, J = 8.00 Hz, 1H), 7.01 (d, J = 7.50 Hz, 1H), 7.17 - 7.25 (m, 1H), 7.56 (dd, J = 8.25, 1.75 Hz, 1H), 7.68 (d, J = 1.75 Hz, 1H), 7.74 (d, J = 8.25 Hz, 1H), 9.43 (s, 1H)

[0582] General procedure for the preparation of compound 6-ET52382-20 [Chemical formula] To a solution of 2-[4-chloro-3-(trifluoromethyl)phenyl]-3-(hydroxymethyl)phenol (30 g, 99.12 mmol) and DIPA (30.09 g, 297.35 mmol, 42.02 mL) in CHCl3 (700 mL) was added 1-bromopyrrolidine-2,5-dione (17.64 g, 99.12 mmol) portionwise at 0 °C. The mixture was warmed to 20 °C and stirred for 12 h. LCMS indicated that 16.1% of the starting material remained and 55% of the product with the desired Ms (Rt = 0.854 min) was detected. One additional reaction was set up as above and the two reaction mixtures were combined. The mixture was poured into water (1000 mL) and extracted with DCM (3 × 300 mL). The organic layers were combined, washed with brine (2 × 100 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography on silica gel (eluting with petroleum ether / ethyl acetate = 50 / 1 to 10 / 1) to afford compound 6 (40 g, 50.24% yield) as a white solid.

[0583] 11H NMR (400 MHz, DMSO-d6) δ 4.08 (br d, J = 3.26 Hz, 2H), 5.11 (br t, J = 4.64 Hz, 1H), 7.03 (d, J = 8.28 Hz, 1H), 7.51 - 7.60 (m, 2H), 7.69 (d, J = 1.88 Hz, 1H), 7.78 (d, J = 8.16 Hz, 1H), 8.93 (br s, 1H)

[0584] General procedure for the preparation of compound 7-ET52382-23

Chem.

[0585] 1 1H NMR (400 MHz, DMSO-d6) δ 4.32 (br d, J = 5.50 Hz, 2H), 7.07 (d, J = 8.38 Hz, 1H), 7.57 - 7.65 (m, 2H), 7.73 (d, J = 1.38 Hz, 1H), 7.84 (d, J = 8.25 Hz, 1H), 9.19 (s, 1H)

[0586] General procedure for the preparation of compound 8-ET52382-25

Chem.

[0587] 1 H NMR (400 MHz, DMSO-d6) δ 4.69 (br s, 2H), 6.75 - 6.85 (m, 2H), 7.06 (d, J = 8.25 Hz, 1H), 7.21 - 7.30 (m, 2H), 7.55 - 7.65 (m, 2H), 7.67 - 7.76 (m, 2H), 9.16 (s, 1H)

[0588] General procedure for the preparation of compound 9-ET52382-33

Chemical Structure

[0589] 1 H NMR (400 MHz, DMSO-d6) δ 3.78 (s, 3H), 4.61 - 4.91 (m, 2H), 6.80 (s, 1H), 6.82 - 6.91 (m, 2H), 7.21 - 7.31 (m, 3H), 7.37 (d, J = 7.88 Hz, 1H), 7.65 (dd, J = 8.19, 1.69 Hz, 1H), 7.71 - 7.80 (m, 2H), 9.01 (s, 1H)

[0590] General procedure for the preparation of compound 10-ET52382-58

Chemical Structure

[0591] 1 H NMR (400 MHz, DMSO-d6) δ 1.71 (s, 3H), 3.82 (s, 3H), 4.80 - 4.99 (m, 2H), 6.78 (s, 1H), 6.84 - 6.91 (m, 2H), 7.24 - 7.32 (m, 2H), 7.61 (br d, J = 7.88 Hz, 1H), 7.72 (br s, 1H), 7.75 (s, 2H), 7.79 (d, J = 8.25 Hz, 1H)

[0592] General procedure for the preparation of compound 11 - ET52382 - 61

Chemical Structure

[0593] Compound 11: 1 H NMR (400 MHz, DMSO-d6) δ 1.65 (s, 3H), 3.73 (s, 3H), 4.72 - 4.97 (m, 2H), 6.82 (br d, J = 8.82 Hz, 2H), 7.22 (br d, J = 8.82 Hz, 2H), 7.47 - 7.70 (m, 2H), 7.70 - 7.80 (m, 3H) Compound 11 by-product: 1 H NMR (400 MHz, DMSO-d6) δ 1.72 (s, 3H), 3.80 (s, 3H), 4.94 - 5.15 (m, 2H), 7.09 (br d, J = 8.88 Hz, 1H), 7.33 (dd, J = 8.88, 2.50 Hz, 1H), 7.56 (d, J = 2.38 Hz, 1H), 7.59 - 7.77 (m, 2H), 7.78 - 7.90 (m, 3H) Separation method for prep-HPLC: Equipment: Shimadzu LC-8A preparative HPLC Column: Phenomenex luna C18 (250 × 70 mm, 15 μm) Mobile phase: A for H2O (0.09% TFA) and B for CAN Gradient: 73% - 90% B in 20 min Flow rate: 130 mL / min Wavelength: 220 and 254 nm

[0594] General procedure for the preparation of A02B01C07D01_P1-ET52382-63

Chemical formula

[0595] 1 H NMR (400 MHz, DMSO-d6) δ 3.77 (s, 3H), 4.78 (br s, 2H), 6.82 - 6.91 (m, 2H), 7.24 - 7.32 (m, 3H), 7.40 (d, J = 8.00 Hz, 1H), 7.64 - 7.70 (m, 1H), 7.72 - 7.76 (m, 1H), 7.78 (s, 1H), 9.24 (s, 1H) LCMS (ESI+): m / z 596.9 (M + H)+, RT: 3.260 min 30_90AB_6 min - 220 - 254 - ELSD: 30 - 90 AB_6 min - 220 - 254 - ELSD: LC / MS (Gradient holds 30% B at 0.40 min, 30 - 90% B at 2.60 min, 90% B for 1.00 min, then 90 - 30% B in 0.01 min, and the flow rate was 1.0 mL / min. Mobile phase A was 0.04% trifluoroacetic acid in water, and mobile phase B was 0.02% trifluoroacetic acid in acetonitrile. The column used for chromatography was Kinetex C18 2.1×50 mm, 5 μm. The detection methods were diode array (DAD) and evaporative light scattering detection (ELSD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0596] General procedure for the preparation of A02B01C07D01_P2 - ET52382 - 64

[0597]

Chem.

[0598] 11H NMR (400 MHz, DMSO-d6) δ 3.78 (s, 3H), 4.90 (br s, 2H), 7.05 (d, J = 8.88 Hz, 1H), 7.26 - 7.37 (m, 2H), 7.39 - 7.47 (m, 1H), 7.55 (d, J = 2.25 Hz, 1H), 7.64 - 7.71 (m, 1H), 7.72 - 7.82 (m, 2H), 9.27 (s, 1H) LCMS (ESI+): m / z 630.9 (M + H)+, RT: 2.670 min 50 - 100 AB_6 min - 220 - 254 - ELSD: LC / MS (Gradient holds 50% B at 0.40 min, 50 - 100% B at 2.60 min, 100% B at 1.00 min, then 100 - 50% B at 0.01 min, and the flow rate was 1.0 mL / min. Mobile phase A was 0.04% trifluoroacetic acid in water and mobile phase B was 0.02% trifluoroacetic acid in acetonitrile. The column used for chromatography was Kinetex C18 2.1×50 mm, 5 μm. The detection methods were diode array (DAD) and evaporative light scattering detection (ELSD). The MS mode was positive electrospray ionization. The MS range was 100 - 1000.)

[0599] Synthesis of NUCC - 0226605 (ET42365 - 477 - 1 and ET42365 - 496 - 1)

Chem.

Chem.

[0600] Experiment for the largest - scale execution: General procedure for the preparation of compound 2 - ET42365 - 393

[0601] The reactions were carried out in parallel but combined for purification.

[0602] [Chemical formula] To a solution of compound 1 (10 g, 43.28 mmol, 1 equiv) in MeOH (100 mL) was added H2SO4 (3.68 g, 37.52 mmol, 2 mL), and the reaction mixture was stirred at 70 °C for 12 h. The reaction mixture was cooled to room temperature, and LCMS indicated that the starting material was consumed and a new peak with the desired product Ms was detected. One more reaction was set up as above, and both reaction mixtures were combined. The combined reaction mixture was concentrated to give a residue, which was diluted with water (100 mL), adjusted to pH = 6 - 7 with aqueous K2CO3, and extracted with ethyl acetate (3×100 mL). The combined organic layers were washed with brine (200 mL), dried over Na2SO4, and concentrated to give compound 2 (21 g, 94.04% yield) as a yellow solid, which was used directly in the next step without further purification.

[0603] 11H NMR (ET42365-393-P1A, 400 MHz, chloroform-d) δ 3.94 (s, 3H), 3.95 (s, 3H), 7.03 (dd, J = 8.03, 1.51 Hz, 1H), 7.26 - 7.30 (m, 1H), 7.31 - 7.37 (m, 1H)

[0604] General procedure for the preparation of compound 3 - ET42365-403

Chemical Structure

Claims

1. A compound having formula I, wherein 【Chemical 1】 in the formula,[[]] W is CR ８ or N, and Y is CH or N; Q is CR ２ or N, and Z is C(Alk ２ ), ｑ (X) ｐ (Alk １ ), ｎ R １ or N, R １ is hydrogen, halo, alkyl, aryl, benzyl, heteroaryl, cycloalkyl, alkoxy or cycloheteroalkyl, and R １ is optionally substituted at one or more positions with one or more of alkyl, alkoxy, cycloalkyl, haloalkyl, haloalkoxy, aryl, hydroxyl, halo, cyano, amide, carboxyl, -(CH ２ )-NH-R ９ , -(O(CH ２ ) ２ ) ｍ -R １５ , or -OR １１ and is optionally substituted at one or more positions, Alk １ is a linear or branched alkenylenyl, or cycloalkenylenyl, and n is 0, 1 or 2; p is 0 or 1; Alk ２ is a linear or branched alkenyl, q is 0 or 1; r is 0 or 1; m is an integer selected from 1 to 20; X is O or NR １３ and R ２ is hydrogen or a halo, R ３ is alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyl, -O-C(O)-alkyl, or halo, and R ４ is hydrogen, halo, amino, alkyl or haloalkyl, or R ４ is aryl or benzyl optionally substituted at one or more ring positions with one or more of alkyl, alkoxy, haloalkyl, haloalkoxy, aryl, hydroxyl, halo, cyano, amide, carboxyl, aryloxy, and alkylaryloxy, or R ４ is alkyl optionally substituted with halo-substituted aryloxy, or R ４ is R １ together with R forms cycloheteroalkyl fused to ring A, and said cycloheteroalkyl fused to ring A is optionally substituted with aryl or alkylaryl optionally substituted with halogen, R ５ is hydrogen, halo, alkyl, aryl, alkylaryl, heteroaryl, cycloalkyl or cycloheteroalkyl, and optionally, R ５ is substituted at one or more positions with one or more of alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyl, halo, cyano, carboxamide, carboxy, aryloxy and heteroaryloxy, R ６ is hydrogen, halo, alkyl, aryl, alkylaryl, heteroaryl, cycloalkyl or cycloheteroalkyl, and optionally, R ６ is substituted at one or more positions with one or more of alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyl, halo, cyano, carboxamide, carboxy, aryloxy and heteroaryloxy, R ７ is alkyl and R ８ is hydrogen, cyano, amino, alkyl, alkoxy, haloalkyl, haloalkoxy, aryl, benzyl, hydroxyl, halo, amide, and carboxyl, R ９ is alkyl optionally substituted with one or more -P(O)(OR １０ ), ２ and is R １０ is hydrogen or alkyl, R １１ is alkyl optionally substituted with one or more -P(O)(OR １２ ), ２ and is R １２ is hydrogen or alkyl, R １３ is hydrogen, alkyl or -C(O)R １４ wherein, R １４ is aryl optionally substituted at one or more ring positions with one or more alkyl, alkoxy, haloalkyl, haloalkoxy, aryl, hydroxyl, halo, cyano, amide and carboxyl, R １５ is OR １６ -OS(O) ２ -R １６ or NR １７ R １８ and R １６ is alkyl or aryl, and R １６ is optionally substituted at one or more positions with one or more of alkyl, alkoxy, haloalkyl, haloalkoxy, aryl, hydroxyl, halo, cyano, amide and carboxyl, R １７ and R １８ are each, independently, hydrogen or alkyl, provided that the compound is not 4'-chloro-6-((4-chlorobenzyl)oxy)-3-(1-methyl-3-(trifluoromethyl)-1H-pyrazol-5-yl)-3'-(trifluoromethyl)-[1,1'-biphenyl]-2-ol.[[]]

2. The compound according to claim 1, having formula I(a). [Chemical 2]

3. R １ is hydrogen, alkyl, aryl, or benzyl, and R １ is optionally substituted at one or more positions with alkyl, alkoxy, aryl, halo, -(CH ２ )-NH-R ９ , or -OR １１ ; R ２ is hydrogen or chloro, R ３ is hydroxyl or -OC(O)Me, and R ４ is hydrogen or a halo, and R ５ is hydrogen or a halogen, and R ６ is hydrogen, aryl, benzyl, and optionally, R ６ is substituted at one or more positions with haloalkyl, halo, and cyano, R ８ is cyano, amino, haloalkyl or amide, and R １４ The compound according to claim 2, wherein R is aryl optionally substituted at one or more ring positions with one or more haloalkyl or halo.

4. Y is N and W is CCF ３ The compound according to claim 3, wherein

5. R ３ The compound according to claim 4, wherein R is hydroxyl.

6. r is 0 and R ６ is phenyl substituted with chloro and trifluoromethyl, the compound according to claim 5.

7. R ２ The compound according to claim 6, wherein R is hydrogen.

8. The compound according to claim 2, selected from the group consisting of [Chemical Formula 3] 【Chemical Formula 4】 【Chemical Formula 5】 【Chemical Formula 6】 【Chemical Formula 7】 【Chemical 8】 【Chemical Formula 9】 【Chemical 10】 【Chemical 11】 [[]]

9. A compound having formula II, wherein 【Chemical 12】 in the formula,[[]] R １ is hydrogen, cyano, amino, alkyl, alkoxy, haloalkyl, haloalkoxy, aryl, benzyl, hydroxyl, halo, amide, and carboxyl, R ２ is alkyl and R ３ is alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyl, -O-C(O)-alkyl, or halo, R ４ is hydrogen, halo, alkyl, aryl, alkylaryl, heteroaryl, cycloalkyl or cycloheteroalkyl, and optionally, R ４ is substituted at one or more positions with one or more of alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyl, halo, cyano, carboxamide, carboxy, aryloxy and heteroaryloxy, X is O or NR ５ and R ５ is hydrogen, alkyl or -C(O)R ６ wherein R ６ is aryl optionally substituted at one or more ring positions with one or more alkyl, alkoxy, haloalkyl, haloalkoxy, aryl, hydroxyl, halo, cyano, amide and carboxyl, Y is alkenylenyl, arylenylenyl, benzylenylenyl, heteroarylenylenyl, cycloalkenylenyl or cycloheteroalkenylenyl; L is a linker selected from the group consisting of a bond, or -O(CH ２ ), ２ ), ａ -NH-C(O)-Alk ２ ), ２ ), ａ -C(O)-NH-((CH ３ ), ２ ), ２ ), ａ -Alk ２ ), ２ -O) ｂ -Alk ３ -C(O)-NH-Alk ２ ), ２ ), ａ -C(O)-NH-Alk ３ -C(O)-, -O(CH ２ ), ２ ), ａ -C(O)-NH-((CH ２ ), ２ -O) ｂ -O(CH ２ ), ２ ), ａ -C(O)-NH-((CH ２ ), ２ -O) ｂ -Alk ３ -C(O)-, -O(CH ２ ), ２ ), ａ -C(O)-NH-Alk ３ -NH-C(O)-CH ２ - and -O(CH ２ ), ２ ), ａ -C(O)-NH-Alk ３ - and is a is an integer selected from 1 to 20; b is an integer selected from 1 to 20; Alk ３ is a linear or branched alkenyl, M Ｅ３ is 【Chemical 13】 selected from the group consisting of[[]] R １１ is a compound that is hydrogen or alkyl.

10. The compound according to claim 9, having the structure of formula II(a): 【Chemical 14】 [[]] in the formula,[[]] R ３ is hydroxyl or -O-C(O)-alkyl, R ４ is hydrogen, or phenyl substituted at one or more positions with haloalkyl or halo, X is O or -N-C(O)R ６ wherein R ６ is aryl substituted at one or more ring positions with one or more haloalkyl or halo, Y is alkenylenyl or benzylenylenyl; L is a linker selected from the group consisting of a bond, or -O(CH ２ ), ２ ), ａ -O(CH ２ ), ２ ), ａ -C(O)-NH-((CH ２ ), ２ -O) ｂ -Alk ３ -O(CH ２ ), ２ ), ａ -C(O)-NH-Alk ３ -C(O)-, -O(CH ２ ), ２ ), ａ -C(O)-NH-((CH ２ ), ２ -O) ｂ -O(CH ２ ), ２ ), ａ -C(O)-NH-((CH ２ ), ２ -O) ｂ -Alk ３ -C(O)-, -O(CH ２ ), ２ ), ａ -C(O)-NH-Alk ３ -NH-C(O)-CH ２ -, and -O(CH ２ ), ２ ), ａ -C(O)-NH-Alk ３ -, and is R ８ is alkyl or aryl, and R ８ is optionally substituted at one or more positions with one or more alkyls, the compound according to claim 9.

11. R ３ is hydroxyl, and R ４ is phenyl substituted with trifluoromethyl and chloro, the compound according to claim 10.

12. X is -N-C(O)R ６ wherein R ６ is phenyl substituted with trifluoromethyl and chloro, the compound according to claim 11.

13. The compound according to claim 11, wherein X is O and Y is propylenylenyl or benzylenylenyl.[[]]

14. The compound according to claim 9, selected from the group consisting of 【Chemical Formula 15】 【Chemical 16】 【Chemical 17】 【Chemical 18】 【Chemical Formula 19】 [[]]

15. A pharmaceutical composition comprising the molecule according to claim 1 and a suitable pharmaceutical carrier, excipient or diluent.[[]]

16. A method for treating cancer, comprising administering the composition according to claim 15 to a subject having said cancer.[[]]

17. The method according to claim 16, wherein the cancer is selected from multiple myeloma, leukemia, non-small cell lung cancer, colon cancer, cancer of the central nervous system, melanoma, ovarian cancer, renal cancer, prostate cancer, and breast cancer.[[]]

18. A pharmaceutical composition comprising the molecule according to claim 9 and a suitable pharmaceutical carrier, excipient or diluent.[[]]

19. A method for treating cancer, comprising administering the composition according to claim 18 to a subject having said cancer.[[]]

20. The method according to claim 19, wherein the cancer is selected from multiple myeloma, leukemia, non-small cell lung cancer, colon cancer, cancer of the central nervous system, melanoma, ovarian cancer, renal cancer, prostate cancer, and breast cancer.[[]]

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