TWI664179B - Substituted 5-(3,5-dimethylisoxazol-4-yl)indoline-2-ones - Google Patents

Abstract

The present invention discloses a substituted 5- (3,5-dimethyliso Azole-4-yl) indol-2one compounds, including at least one 4-substituted 5- (3,5-dimethyliso Azol-4-yl) indoline-2 ketone pharmaceutical composition, its preparation method, and its use for inhibiting the BET family of bromo domains and treating diseases caused thereby, such as specific cancers.

Description

Substituted 5- (3,5-dimethylisoazol-4-yl) indol-2-one

The present invention is directed to methods and compounds for inhibiting BRD4 and treating diseases associated with undesired BRD4 activity.

One of the most frequently occurring post-translational modifications in proteins is ε-N-acetylation of lysine residues (Choudhary et al., 2009), and it is widely related to cell communication and disease biology. In the macromolecular complexes involved in chromatin remodeling, DNA damage repair, and cell cycle regulation, a large amount of acetylated lysine is present in histones (Choudhary et al., 2009). Targeted chromatin-modifying enzymes that control the degree of cell acetylation, called epigenetic "writers" (Histone Acetyltransferases, HATs) and "eraser" (Histone deacetylases, HDACS) have been developed There has been large-scale research in the field of drug development, but "readers" (bromodomains) that regulate recognition of acetylation sites have not been widely reported until recently (Nicodeme et al. 2010, Filippakopoulos et al. 2010). The Bromo domain (BRDs) family is a class of evolutionarily conserved modules of protein interactions that can specifically recognize acetylated lysine-containing protein motifs. The extra-terminal (BET) bromo domain subfamily includes BRD2, BRD3, BRD4, and BRDT, two amine-terminal bromo domains with high sequence homology, and a carboxy-terminal domain Common structural features. Recent studies have confirmed the use of the BET Bromo domain as a target for the treatment of multiple cancers (Filippakopoulos 2010, Delmore 2011, Zuber 2011), atherosclerosis (Chung 2011, Mirguet 2012), inflammation (Nicodeme 2010), and HIV infection (Banerjee 2012).

MYC transcription factors are important regulators of various cellular functions, and have long been proven to be a series of attractive human cancer therapeutic targets. However, strategies for regulating the functions of Myc oncoproteins have not yet been discovered. Recently, two selective inhibitors of the BET family members (with little activity on Bromo domains outside the BET family), the JQ1 and I-BET151 lines have been shown to strongly downregulate MYC protein and MYC target gene transcription. JQ1 and IBET-151 strongly inhibit the tumor growth of multiple myeloma in vivo and in vitro in a MYC-dependent manner, a variety of leukemia and lymphoma cell lines, and samples from patients with primary leukemia (Delmore 2011, Mertz, 2011, Zuber 2011, Herrmann 2010 , Dawson 2011). BET Bromo domain inhibitors can also be used to treat other MYC-dependent cancers, such as neuroblastoma with MYCN amplification and other solid tumors with over-represented c-MYC. In addition, JQ1 has also been shown to be incurable The human squamous cell carcinoma subtype is known to have antiproliferative effects in NUT midline cancer (NMC). NMC is generally defined as the tandem N-terminal Bromo domain of BRD4 or BRD3 caused by t (15,19) chromosome rearrangement and forms an in-frame chimera with NUT (testicular nucleoprotein) protein. JQ1 treatment resulted in the final differentiation, cell cycle arrest, and apoptosis of NMC cell lines, and resulted in significantly slower tumor growth from patient xenograft (Filippakopoulos 2010). In summary, BET bromo domain inhibitors can be used to treat a variety of human cancers.

IBET, a BET bromo domain inhibitor, has been reported to inhibit several important pro-inflammatory cytokines and chemokines by replacing the BET protein on the inflammatory gene promoter. The IBET line showed anti-inflammatory effects and prevented endotoxin-induced animal death in the sepsis model of mice (Nicodeme 2010). These studies indicate that the BET bromo domain system can be used in immunomodulatory drugs.

The up-regulated system of lipoprotein A1 (ApoA1) is associated with the prevention of atherosclerosis progression and also has an anti-inflammatory effect (Nicholls 2012). Studies have found that BET bromo domain inhibitors can increase the performance of ApoA1. BET inhibition is a promising therapy for the development of new therapies for atherosclerosis.

The persistence of latent HIV-1 is a major challenge for the complete eradication of infection. JQ1 has been reported to be able to reactivate HIV in a pattern of latent T-cell infection and latent monocyte infection (Banerjee 2012, Li 2003). Combination therapy with BET Bromo domain inhibitors for reactive latent HIV-1 viruses can provide an opportunity to cure HIV-1 infection. Other BET bromo domain inhibitors are known, see for example WO2012151512, WO2012143436, WO2012075383, WO2011161031, WO2011143669, WO2011143657, WO2011054848, WO2011054846, WO2011054845, WO2011054844, WO2011054843, WO2011054841, WO2011054553, WO2009084693 and WO2006032470. Specific 3,5-dimethyliso The azoles have been identified as BRD4 inhibitors (David S. Hewings 2011).

Related Literature

Filippakopoulos, Panagis, Jun Qi, Sarah Picaud, Yao Shen, William B. Smith, Oleg Fedorov, Elizabeth M. Morse et al. "Selective inhibition of BET bromodomains." Nature 468, no. 7327 (2010): 1067-1073

Delmore, Jake E., Ghayas C. Issa, Madeleine E. Lemieux, Peter B. Rahl, Junwei Shi, Hannah M. Jacobs, Efstathios Kastritis et al. "BET bromodomain inhibition as a therapeutic strategy to target c-Myc." Cell 146, no. 6 (2011): 904-917.

Zuber, Johannes, Junwei Shi, Eric Wang, Amy R. Rappaport, Harald Herrmann, Edward A. Sison, Daniel Magoon et al. "RNAi screen identifies Brd4 as a therapeutic target in acute myeloid leukaemia." Nature 478, no. 7370 ( 2011): 524-528.

Chung, Chun-wa, Hervé Coste, Julia H. White, Olivier Mirguet, Jonathan Wilde, Romain L. Gosmini, Chris Delves et al. "Discovery and characterization of small molecule inhibitors of the BET family bromodomains." Journal of medicinal chemistry 54 , no. 11 (2011): 3827-3838.

Dawson, Mark A., Rab K. Prinjha, Antje Dittmann, George Giotopoulos, Marcus Bantscheff, Wai-In Chan, Samuel C. Robson et al. "Inhibition of BET recruitment to chromatin as an effective treatment for MLL-fusion leukaemia." Nature 478, no. 7370 (2011): 529-533. Nicodeme, Edwige, Kate L. Jeffrey, Uwe Schaefer, Soren Beinke, Scott Dewell, Chun-wa Chung, Rohit Chandwani et al. "Suppression of inflammation by a synthetic histone mimic. " Nature 468, no. 7327 (2010): 1119-1123.

Hewings, David S., Minghua Wang, Martin Philpott, Oleg Fedorov, Sagar Uttarkar, Panagis Filippakopoulos, Sarah Picaud et al. "3, 5-Dimethylisoxazoles act as acetyl-lysine-mimetic bromodomain ligands." Journal of medicinal chemistry 54, no 19 (2011): 6761-6770.

Choudhary, C., Kumar, C., Gnad, F., Nielsen, ML, Rehman, M., Walther, TC, Olsen, JV, and Mann, M. (2009). Lysine acetylation targets protein complexes and co-regulates major cellular functions. Science 325, 834-840.

Herrmann H, Blatt K, Shi J, Gleixner KV, Cerny-Reiterer S, Müllauer L, Vakoc CR, Sperr WR, Horny HP, Bradner JE, Zuber J, Valent P Small-molecule inhibition of BRD4 as a new potent approach to eliminate leukemic stem- and progenitor cells in acute myeloid leukemia AML Oncotarget. 2012 Nov 27. [Epub ahead of print]

Mertz, Jennifer A., Andrew R. Conery, Barbara M. Bryant, Peter Sandy, Srividya Balasubramanian, Deanna A. Mele, Louise Bergeron, and Robert J. Sims III. "Targeting MYC dependence in cancer by inhibiting BET bromodomains." Proceedings of the National Academy of Sciences 108, no. 40 (2011): 16669-16674.

Banerjee C, Archin N, Michaels D, Belkina AC, Denis GV, Bradner J, Sebastiani P, Margolis DM, Montano M. BET bromodomain inhibition as a novel strategy for reactivation of HIV-1. J Leukoc Biol. 2012 Dec; 92 ( 6): 1147-54.

Li, Zichong, Jia Guo, Yuntao Wu, and Qiang Zhou. "The BET bromodomain inhibitor JQ1 activates HIV latency through antagonizing Brd4 inhibition of Tat-transactivation." Nucleic Acids Research 41, no. 1 (2013): 277-287.

Chung, Chun-wa, Hervé Coste, Julia H. White, Olivier Mirguet, Jonathan Wilde, Romain L. Gosmini, Chris Delves et al. "Discovery and characterization of small molecule inhibitors of the BET family bromodomains." Journal of medicinal chemistry 54 , no. 11 (2011): 3827-3838.

Mirguet, Olivier, Yann Lamotte, Frédéric Donche, Jérôme Toum, Françoise Gellibert, Anne Bouillot, Romain Gosmini et al. "From ApoA1 upregulation to BET family bromodomain inhibition: Discovery of I-BET151." Bioorganic & medicinal chemistry letters (2012).

Nicholls, Stephen J., Allan Gordon, Jan Johannson, Christie M. Ballantyne, Philip J. Barter, H. Bryan Brewer, John JP Kastelein, Norman C. Wong, Marilyn RN Borgman, and Steven E. Nissen. "ApoA-I Induction as a Potential Cardioprotective Strategy: Rationale for the SUSTAIN and ASSURE Studies. "Cardiovascular drugs and therapy (2012): 1-7.

David S. Hewings, Minghua Wang, Martin Philpott, Oleg Fedorov, Sagar Uttarkar, Panagis Filippakopoulos, Sarah Picaud, Chaitanya Vuppusetty, Brian Marsden, Stefan Knapp, Stuart J. Conway and Tom D. Heightman. 3,5 Dimethylisoxazoles Act As Acetyl- lysine-mimetic Bromodomain Ligands. J. Med. Chem. 2011, 54, 6761-6770.

The present invention provides methods and compositions for inhibiting BRD ₄ and treating diseases associated with undesired BRD ₄ activity.

In a specific example, the present invention provides a BRD ₄ inhibitor or compound of the formula:

Its stereoisomers, and their pharmaceutically acceptable salts, of which:

R ¹ is hydrogen, halide, heteroatom functional group or hydrocarbon group, and the hydrocarbon group is selected from C1-C8 alkyl group, C2-C8 alkenyl group, C2-C8 alkynyl group and C6-C14 aryl group. The alkynyl and alkynyl systems are selectively cyclized, and each hydrocarbyl system is selectively substituted, and optionally contains 1 to 3 heteroatoms.

R ² is a heteroatom functional group or a hydrocarbon group. The hydrocarbon group is selected from C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, and C6-C14 aryl. Among them, each alkyl, alkenyl, and alkynyl group is selective Upon cyclization, each hydrocarbyl system is selectively substituted and optionally contains 1 to 3 heteroatoms.

R ³ is halogen, lower alkyl, hydroxy, lower alkoxy, or lower fluorenyl, and n is 0, 1, 2, or 3.

Each combination of the present invention has been enumerated in detail, including all the combinations of the specific examples listed.

In the specific examples exemplified: R ¹ may be a halide, -OR ⁴ or -NR ⁵ R ⁶ , or a hydrocarbon group selected from a C1-C8 alkyl group, a C2-C8 alkenyl group, a C2-C8 alkynyl group, and C6-C14 aryl, in which each alkyl, alkenyl, alkynyl group is selectively cyclized, each hydrocarbon group is selectively substituted, and optionally contains 1 to 3 heteroatoms, of which R ⁴ , R ⁵ and Each of R ⁶ is H or a hydrocarbon group. The hydrocarbon group is selected from C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, and C6-C14 aryl. Among them, each alkyl, alkenyl and alkynyl are selected. Cyclization, and each hydrocarbon group is optionally substituted, and optionally contains 1 to 3 heteroatoms, in which R ⁵ and R ⁶ atoms connected together, each form a selectively substituted cyclic hydrocarbon ring; or R ¹ is -OR ⁴ or -NR ⁵ R ⁶ , or a hydrocarbon group, and the hydrocarbon group may be selected from C3-C8 cycloalkyl, C5-C8 cycloalkenyl, and C6-C14 aryl, wherein each hydrocarbon group is optionally substituted, and Optionally contains 1 to 3 heteroatoms, wherein each of R ⁴ , R ⁵ and R ⁶ is H or a hydrocarbon group, the hydrocarbon group is selected from C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, and C6- C14 aryl, among which alkyl, alkenyl and alkynyl are selective Cyclization, and various hydrocarbon group is selectively substituted, and selectively contains 1 to 3 heteroatoms, wherein R ⁵ and R ⁶ atoms connected thereto together form the respective selectively substituted cycloalkyl ring; or R ¹ is -OR ⁴ or -NR ⁵ R ⁶ , or a C6-C14 aryl group, wherein the aryl group is optionally substituted and optionally contains 1 to 3 heteroatoms, and R ⁴ , R ⁵ and R ⁶ are each H or a hydrocarbyl group, the hydrocarbyl group is selected from C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, and C6-C14 aryl, wherein each alkyl, alkenyl and alkynyl are selectively cyclized, and Each hydrocarbyl group is optionally substituted, and optionally contains 1 to 3 heteroatoms, wherein R ⁵ or R ⁶ is an atom to which it is attached together, each forming a selectively substituted cyclic hydrocarbyl ring.

In the specific examples exemplified: R ² is -OR ⁴ or -NR ⁵ R ⁶ , or a hydrocarbon group, and the hydrocarbon group is selected from C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, and C6-C14 aromatic Group, wherein each alkyl group, alkenyl group and alkynyl group are selectively cyclized, and each hydrocarbon group is selectively substituted, and optionally contains 1 to 3 heteroatoms, wherein R ⁴ , R ⁵ and R ⁶ are each Is H or a hydrocarbyl group, the hydrocarbyl group is selected from a C1-C8 alkyl group, a C2-C8 alkenyl group, a C2-C8 alkynyl group, and a C6-C14 aryl group, wherein each alkyl group, alkenyl group, and alkynyl group are selectively cyclized, Each hydrocarbyl group is optionally substituted and optionally contains 1 to 3 heteroatoms, in which R ⁵ and R ⁶ are atoms bonded together to form a selectively substituted cyclic hydrocarbyl ring; or R ² is -NR ⁵ R ⁶ , Or a hydrocarbon group, the hydrocarbon group is selected from the group consisting of C3-C8 cycloalkyl, C5-C8 cycloalkenyl, and C6-C14 aryl, wherein each hydrocarbon group is optionally substituted and optionally contains 1 to 3 heteroatoms, where R ⁴ , R ⁵ and R ⁶ are each H or a hydrocarbon group, and the hydrocarbon group is selected from C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, and C6-C14 aryl, wherein each alkyl, alkenyl and Alkynyl is selectively cyclized, each hydrocarbon is selectively substituted And optionally contains 1 to 3 heteroatoms, in which R ⁵ and R ⁶ are atoms bonded together to form a selectively substituted cycloalkyl ring; or R ² is -NR ⁵ R ⁶ , or C6-C14 aryl Wherein, the aryl group is optionally substituted and optionally contains 1 to 3 heteroatoms, and R ⁴ , R ⁵ and R ⁶ are each H or a hydrocarbon group, and the hydrocarbon group is selected from C1-C8 alkyl and C2-C8 Alkenyl, C2-C8 alkynyl, and C6-C14 aryl, wherein each alkyl, alkenyl, and alkynyl are selectively cyclized, and each hydrocarbyl is selectively substituted, and optionally contains 1 to 3 A heteroatom, in which R ⁵ and R ⁶ are connected together, to form a selectively substituted cyclic hydrocarbyl ring.

In the specific examples exemplified, the present invention provides the compounds of Tables 1 and 2 or the examples herein, and pharmaceutically acceptable salts thereof.

Another aspect of the present invention provides a substituted 5- (3,5-dimethyliso Azole-4-yl) indoline-2-one, a BRD ₄ -inhibitor, and in the specific example exemplified, the inhibitor includes an indoline with 1 to 2 substituents at the C3 position Compound. These substituents may be a halide, a heteroatom functional group, or a hydrocarbon group, respectively. The hydrocarbon group is selected from C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, and C6-C14 aryl, wherein each alkyl group Alkenyl and alkynyl are selectively cyclized, and each hydrocarbyl is selectively substituted, and optionally contains 1 to 3 heteroatoms.

The present invention also provides the title compound with BRD ₄ -inhibitory activity, which has an IC50 value of 10 μM or less in the BRD ₄ time-resolved fluorescence resonance energy transfer (TR-FRET) enzyme analysis measurement, and the analysis is based on the Bromo structure the mixture field, the compounds and tetra acetylated histone peptide used (E. coli) expression and purification from E. coli having N- terminal His identified recombinant human BRD4 (1-477) bromodomain.

The invention also provides a pharmaceutical composition comprising a therapeutically effective amount of the title compound in unit dosage form, and one or more pharmaceutically acceptable carriers.

The invention also provides a therapeutically effective amount of the title compound and a different agent having anticancer therapeutic activity.

The invention also provides a method of treating a disease associated with undesired BRD ₄ activity, the method comprising administering to a human in need thereof an effective dose of the title compound, its N-oxide, or prodrug, wherein the disease is specifically cancer.

The invention also provides pharmaceutical compositions, including the title compound in unit dosage and edible form, and methods for inducing autophagy, including administering an effective amount of the title compound or composition to a person in need thereof.

The present invention also provides the use of the title compound as a medicament, as well as the use of a medicament for the manufacture of a medicament for treating a disease associated with an undesired BDR ₄ activity.

The present invention discloses novel compounds that inhibit BET regions such as BRD4.

Unless the context indicates otherwise, the words, phrases, and symbols used below are generally intended to have the following meanings. The following abbreviations and terms have meanings throughout the text.

The term "alkyl" refers to a hydrocarbyl group selected from saturated straight and branched chain hydrocarbyl groups including 1 to 18, or 1 to 12, or 1 to 6 carbon atoms. Examples of alkyl include methyl, ethyl, 1-propyl or n-propyl ("n-Pr"), 2-propyl or isopropyl ("i-Pr"), 1-butyl or n- Butyl ("n-Bu"), 2-methyl-1-propyl or isobutyl ("i-Bu"), 1-methylpropyl or second butyl ("s-Bu"), and 1,1-dimethylethyl or third butyl ("t-Bu"). Examples of other alkyl groups include 1-pentyl, N-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1 -Butyl, 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl 3-pentyl, 2-methyl-3-pentyl, 2,3-dimethyl-2-butyl and 3,3-dimethyl-2-butyl groups.

Lower alkyl means 1-8, preferably 1-6, more preferably 1-4 carbon atoms; lower alkenyl or alkynyl means 2-8, 2-6 or 2-4 carbon atoms.

The term "alkenyl" refers to a hydrocarbon group selected from straight and branched chain hydrocarbon groups including at least one C = C double bond and 2-18, or 2-12, or 2-6 carbon atoms. Examples of alkenyl may be selected from ethyl or vinyl, prop-1-enyl, prop-2-enyl, 2-methylprop-1-enyl, but-1-enyl, but-2 -Alkenyl, but-3-enyl, but-1,3-dienyl, 2-methylbut-1,3-dienyl, hex-1-enyl, hex-2-enyl, hex 3-alkenyl, hex-4-enyl and hex-1,3-dienyl groups.

The term "alkynyl" refers to a hydrocarbon group selected from straight and branched chain hydrocarbon groups including at least one C≡C triple bond and 2-18, or 2-12, or 2-6 carbon atoms. Examples of alkynyl include ethynyl, 1-propynyl, 2-propynyl (propargyl), 1-butynyl, 2-butynyl, and 3-butynyl groups.

The term "cycloalkyl" refers to a cyclic hydrocarbon group selected from saturated and partially unsaturated cyclic hydrocarbon groups including monocyclic and polycyclic (e.g., bicyclic and tricyclic) groups. For example, the cycloalkyl group may include 3-12, or 3-8, or 3-6 carbon atoms. For further example, the cycloalkyl group may be a monocyclic group of 3-12, or 3-8, or 3-6 carbon atoms. Examples of monocyclic cycloalkane groups include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopentane-1-enyl, 1-cyclopentane-2-enyl, 1-cyclopentane-3- Alkenyl, cyclohexyl, 1-cyclohexyl-1-enyl, 1-cyclohexyl-2-enyl, 1-cyclohexyl-3-enyl, cyclohexadienyl, cycloheptyl, cyclooctyl, Cyclononyl, cyclodecyl, cycloundecyl, and cyclododecyl. Examples of bicyclic cycloalkyl include 7-12 A bicyclic group or a bridged bicyclic group composed of an arrangement of two ring atoms, the bicyclic ring is selected from [4,4], [4,5], [5,5], [5,6], and [6,6] ring systems, The bridged bicyclic ring is selected from bicyclic [2.2.1] heptane, bicyclic [2.2.2] octane, and bicyclic [3.2.2] nonane. The ring may be saturated or have at least one double bond (ie, partially unsaturated), but is not fully conjugated, and is not aromatic as defined herein.

The term "aromatic" refers to a group selected from: 5 and 6-membered carbocyclic aromatic rings, for example, phenyl; bicyclic systems such as 7-12 membered bicyclic systems, where at least one ring is a carbocyclic and aromatic ring, Such a bicyclic ring system is selected from, for example, naphthalene, indene and 1,2,3,4-tetrahydroquinoline; and a tricyclic ring system such as a 10-15 member tricyclic ring system, wherein at least one ring is a carbocyclic ring and an aromatic ring, Ruo.

For example, the aryl group is an aryl group selected from the group consisting of a 5- and 6-membered carbocyclic aromatic ring fused to a 5-7-membered cycloalkyl or heterocyclic ring, and the 5-7-membered cycloalkyl or heterocyclic ring Containing at least one heteroatom selected from N, O, and S, if the carbocyclic ring is fused with a heterocyclic ring, the point of attachment may be on the carboaromatic ring, and if the carboaromatic ring is fused with a cycloalkyl group, The point of attachment may be on a carbocyclic or cycloalkyl. A divalent group formed from a substituted phenyl derivative and having a free valence in a ring atom is referred to as a substituted phenylene radical. The monovalent polycyclic hydrocarbon group ends with a "-group". The monovalent polycyclic hydrocarbon group is removed from a carbon atom with a free valence by a hydrogen atom to obtain a divalent group. The name is to add "extended" to the name of the corresponding monovalent group. For example, a naphthyl group having two attachment points is referred to as a dihydronaphthyl group. However, aryl is not included or overlaps with heteroaryl in any case, and will be defined separately below. Therefore, if one or more carbocyclic rings are fused with a heterocyclic aromatic ring, the resulting ring system is a heteroaryl group as defined in the present specification, rather than an aryl group.

The term "halogen" or "halogen" refers to F, Cl, Br or I.

The term "heteroalkyl" refers to an alkyl group that includes at least one heteroatom.

The term "heteroaryl" is selected from: 5- to 7-membered aromatic monocyclic rings containing 1, 2, 3, or 4 heteroatoms selected from N, O, and S, and the remaining ring atoms being carbon; 8 members The 12-membered bicyclic ring includes 1, 2, 3, or 4 heteroatoms selected from N, O, and S. The remaining ring atoms are carbon, and at least one of the rings is aromatic, and at least one of the aromatic rings has One heteroatom; 11 to 14-membered tricyclic rings include 1, 2, 3, or 4 heteroatoms selected from N, O, and S, the remaining ring atoms are carbon and at least one of the rings is aromatic, and aromatic There is at least one heteroatom in the ring.

For example, heteroaryl includes 5 to 7 membered heterocyclic aromatic rings fused to 5 to 7 membered cycloalkyl rings. For such fused rings, only one of the rings in the bicyclic heteroaryl ring system contains at least one heteroatom, and the point of attachment may be a heteroaryl ring or a cycloalkyl ring.

When the total number of S and O atoms on a heteroaryl group exceeds 1, these heteroatoms will not be adjacent. In some specific examples, the total number of S and O on the heteroaryl group does not exceed two. In some specific examples, the total number of S and O on the heteroaromatic ring does not exceed one.

Examples of heteroaryl include, but are not limited to (coded from the preferentially designated attachment position 1) pyridyl (eg 2-pyridyl, 3-pyridyl, 4-pyridyl), Phosphono , 2,4-pyrimidinyl, 3,5-pyrimidinyl, 2,4-imidazolyl, imidazopyridyl, iso Azolyl, Oxazolyl, thiazolyl, isothiazolyl, thiadiazole, tetrazolyl, thienyl, tri , Benzothienyl, furanyl, benzofuranyl, benzimidazolyl, indolyl, isoindolyl, dihydroindolyl, phthaloyl Base Base Base, pyrrolyl, triazolyl, quinolinyl, isoquinolinyl, pyrazolyl, pyrrolopyridyl (such as 1H-pyrrolo [2,3-b] pyridin-5-yl), pyrazolopyridine (Such as 1H-pyrazolo [3,4-b] pyridin-5-yl), benzo Azolyl (such as benzo [d] Azole-6-yl), pyridinyl, purinyl, 1-oxa-2,3-diazolyl, 1-oxa-2,4-diazolyl, 1-oxa-2,5-di Oxazolyl, 1-oxa-3,4-diazolyl, 1-thio-2,3-diazolyl, 1-thio-2,4-diazolyl, 1-thio-2,5 -Diazolyl, 1-thio-3,4-diazolyl, furazyl, benzofurazyl, benzothienyl, benzothiazyl, benzo Oxazolyl Phenyl, naphthyridinyl, furanopyridyl, benzothiazolyl (such as benzo [d] thiazol-6-yl), indolyl (such as 1H-indazol-5-yl), and 5,6,7 , 8-tetrahydroisoquinoline.

The term "heterocyclic" or "heterocyclic" or "heterocyclyl" refers to monocyclic, bicyclic, tricyclic rings selected from 4- to 12-membered, which are saturated and partially unsaturated rings, including In addition to 1, 2, 3 or 4 heteroatoms selected from oxygen, sulfur and nitrogen, at least one carbon atom. "Heterocycle" also refers to a 5- to 7-membered heterocyclic ring consisting of at least one heteroatom selected from N, O, and S, and a 5-, 6-, and / or 7-membered cycloalkyl ring, carbon A ring aromatic ring or a heteroaromatic ring is fused, when the heterocyclic ring is fused with a carbocyclic aromatic ring or a heteroaromatic ring, the connection point is on the heterocyclic ring, and when the heterocyclic ring is fused with a cycloalkyl group The point may be on a cycloalkyl or heterocyclic ring.

"Heterocycle" also refers to an aliphatic spiro ring that includes at least one heteroatom selected from N, O, and S, and the point of attachment is on the heterocycle. The above rings may be saturated or contain at least one double bond (ie, partially unsaturated). The above heterocycle may be substituted with pendant oxygen, and the point of attachment may be a carbon or heteroatom on the heterocycle. Heterocyclic is not a heteroaryl group as defined herein.

Examples of heterocycles include, but are not limited to (coded from the preferred attachment position 1) 1-pyrrolidinyl, 2-pyrrolidinyl, 2,4-imidazolidinyl, 2,3-pyrazolidyl, 1 -Piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-piperidinyl, 2,5-piperidinyl Methyl, piperanyl, 2- Porphyrinyl, 3- Porphyrinyl, oxiranyl, azetidinyl, epithioethane, azetidinyl, oxetanyl, thiacyclobutyl, 1,2-dithiacyclobutyl , 1,3-dithiocyclobutyl, dihydropyridyl, tetrahydropyridyl, thio Oxoline Base Group, homopiperidinyl, azacycloheptyl, oxecanyl, thietyl, 1,4-oxetane, 1,4-dioxetane , 1,4-oxetane, 1,4-azathioheptyl, 1,4-dithioheptyl, 1,4-azathioheptyl and 1,4-diazacycloheptane, 1,4-dithiaalkyl, 1,4-azathiocyclohexyl, oxazepine, diazepine, thiazepine, dihydrothiophene , Dihydropiperanyl, dihydrofuranyl, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropiperanyl, tetrahydrothyranyl, 1-pyrrolidinyl, 2-pyrrolinyl, 3-pyrrolinyl , Dihydroindolyl, 2H-piperanyl, 4H-piperanyl, 1,4-dioxetanyl, 1,3-dioxocyclyl, pyrazolinyl, pyrazolinyl , Dithiaalkyl, dithiacyclopentyl, pyrazolidinyl imidazolinyl, pyrimidinone, 1,1-dioxo-thio Phenyl, 3-azabicyclo [3.1.0] hexyl, 3-azabicyclo [4.1.0] heptyl, and azabicyclo [2.2.2] hexyl. Substituted heterocyclic groups also include ring systems substituted with one or more pendant oxygen groups, such as N-oxidized piperidinyl, N-oxidized Phenyl, 1-oxo-1-thioxo Phenyl and 1,1-dioxo-1-thio Porphyrinyl.

The substituent is selected from: halogen, -R ', -OR', = O, = NR ', = N-OR', -NR'R ", -SR ', halogen, -SiR'R"R'",- OC (O) R ', -C (O) R', -CO ₂ R ', -CONR'R ", -OC (O) NR'R", -NR "C (O) R', -NR ' -C (O) NR "R '", -NR'-SO ₂ NR'",-NR" CO ₂ R ', -NH-C (NH ₂ ) = NH, -NR'C (NH ₂ ) = NH , -NH-C (NH ₂ ) = NR ', -S (O) R', -SO ₂ R ', -SO ₂ NR'R ", -NR" SO ₂ R, -CN and -NO ₂ ,- N ₃ , -CH (Ph) ₂ , perfluoro (C1-C4) alkoxy and perfluoro (C1-C4) alkyl, the number of substituents is from 0 to 3, especially the number of substituents is 0, 1 or 2 groups. R ', R "and R'" are each independently selected from hydrogen, unsubstituted (C1-C8) alkyl and heteroalkyl, unsubstituted aryl, aryl substituted with 1 to 3 halogens, unsubstituted Substituted alkyl, alkoxy or thioalkoxy, or aryl- (C1-C4) alkyl. When R 'and R "are attached to the same nitrogen atom, they can form a 5-, 6-, or 7-membered ring together with the nitrogen atom. Therefore, -NR'R" includes 1-pyrrolidinyl and 4- Phenolinyl, "alkyl" includes, for example, trihaloalkyl (such as -CF ₃ and -CH ₂ CF ₃ ), and when the aryl is 1,2,3,4-tetrahydronaphthalene, it may be substituted or unsubstituted Substituted by substituted (C3-C7) spirocycloalkyl. (C3-C7) spirocycloalkyl may be substituted in the manner defined in this patent as "cycloalkyl".

Preferred substituents are selected from: halogen, -R ', -OR', = O, -NR'R ", -SR ', -SiR'R"R'", -OC (O) R ', -C (O) R ', -CO ₂ R', -CONR'R ", -OC (O) NR'R", -NR "C (O) R ', -NR" CO ₂ R', -NR'- SO ₂ NR "R '", -S (O) R', -SO ₂ R ', -SO ₂ NR'R ", -NR" SO ₂ R, -CN and -NO ₂ , perfluorinated (C1-C4 ) Alkoxy and perfluoro (C1-C4) alkyl, where R 'and R "are as defined above.

The term "fused ring" refers to a polycyclic ring system, for example, a bicyclic or tricyclic ring system in which two rings share only two ring atoms and one bond. Examples of the fused ring may include a fused bicyclic alkyl ring such as a bicyclic ring composed of 7 to 12 ring atoms, the bicyclic ring selected from [4,4], [ 4,5], [5,5], [5,6], and [6,6] ring systems; fused bicyclic aromatic rings, such as the 7 to 12-membered bicyclic aromatic ring system described above, fused tricyclic aromatic rings Rings, such as the 10 to 15-membered tricyclic aromatic ring system described above; fused bicyclic heteroaromatic rings, such as the 8- to 12-membered bicyclic heteroaromatic ring system described above, fused tricyclic heteroaromatic rings , Such as 11- to 14-membered tricyclic heteroaromatic ring systems as described above; and fused bicyclic or tricyclic heteroaromatic rings as described above.

The compounds may contain an asymmetric center and may therefore exist as enantiomers. When compounds have two or more asymmetric centers, they can additionally exist as diastereomers. Enantiomers and diastereomers belong to a broad class of stereoisomers. All these possible stereoisomers include substantially pure resolved enantiomers, racemic mixtures, and diastereomeric mixtures. All stereoisomers of the compounds and / or pharmaceutically acceptable salts thereof are included. Unless otherwise specifically mentioned, one isomer mentioned applies to any reasonable isomer. Whenever the isomeric component is not specified, all possible isomers are included.

The term "substantially pure" means that the target stereoisomer contains other stereoisomers with a weight of not more than 35%, such as not more than 30%, further such as not more than 25%, or even not more than 20%. In some specific examples, the term "substantially pure" means that the target stereoisomer contains other stereoisomers with a weight of not more than 10%, such as not more than 5%, such as not more than 1%.

When the compound in question contains an olefinic double bond, these double bonds include E and Z geometric isomers unless otherwise specified in detail.

Some of the compounds mentioned may have different points of attachment of hydrogen atoms and are called tautomers. For example, compounds including carbonyl -CH ₂ C (O) - group of the compound (keto forms) may undergo tautomerism to form hydroxyl -CH = C (OH) - groups (enol forms). Both keto and enol forms that can be implemented alone and mixtures thereof are also included.

It is advantageous to separate the reaction products from each other and / or from the starting materials. The isolation and / or purification of the desired product in each step or series of steps (hereinafter, isolation is used) is to achieve the desired homogeneity by techniques commonly used in the art. Typically these separations involve heterogeneous extraction, crystallization from a solvent or solvent mixture, distillation, sublimation, or chromatography. Chromatography can involve any number of methods including, for example: reversed phase and normal phase; molecular sieves, ion exchange, high, medium and low pressure liquid chromatography and equipment; small scale analytical; simulated moving bed (" SMB ") and preparative thin-layer or thick-layer chromatography, and small-scale thin-layer and flash chromatography techniques. Those skilled in the art may use these techniques to achieve the desired resolution.

Diastereomeric mixtures can be separated into their respective diastereomers using techniques that are well known in the art, using their physical and chemical differences, such as by chromatography and / or fractional crystallization. Enantiomers can be converted to diastereomeric mixtures by reacting a mixture of enantiomers with a suitable optically active compound (e.g., a chiral auxiliary such as a chiral alcohol or Mosher acid chloride) , Separating the mixture of diastereomers, and Each diastereomer is converted (e.g., hydrolyzed) to the corresponding pure enantiomer. Enantiomers can also be separated by a chiral HPLC column.

Single stereoisomers (e.g., substantially pure enantiomers) can be obtained by resolving racemic mixtures, such as the use of optically active resolving agents to form diastereomers (Eliel, E., and Wilen, S. Stereochemistry of Organic Compounds. New York: John Wiley & Sons, Inc., 1994; Lochmuller, CH, et al. "Chromatographic resolution of enantiomers: Selective review." J. Chromatogr., 113 (3) (1975) ): Pp.283-302). The racemic mixture of the chiral compound of the present invention can be separated by any suitable method, including: (1) forming an ionic, diastereomeric salt with the chiral compound, and then by fractional crystallization or other Method separation, (2) formation of diastereomeric compounds with chiral derivatization reagents, separation of the formed diastereomers and conversion to pure stereoisomers, (3) separation directly under chiral conditions Pure or rich stereoisomers. See: Wainer, Irving W., Ed. Drug Stereochemistry: Analytical Methods and Pharmacology. New York: Marcel Dekker, Inc., 1993.

"Pharmaceutically acceptable salt" includes, but is not limited to, inorganic acid salts selected from, for example, hydrochloride, phosphate, hydrogen phosphate, hydrobromide, sulfate, sulfite, and nitrate; also includes organic Salt selected from, for example, malate, maleate, fumarate, tartrate, succinate, citrate, lactate, methanesulfonate, p-toluenesulfonate, 2-hydroxyethylsulfonate , Benzoates, salicylates, stearates, alkanoates such as acetates, and salts of HOOC- (CH ₂ ) _n -COOH, where n is selected from 0 to 4. Similarly, examples of cations acceptable in medicine include, but are not limited to, sodium, potassium, calcium, aluminum, lithium, and ammonium salts.

Alternatively, if a compound described herein gets its acid addition salt, its free base can be obtained by basifying its salt solution. Conversely, if the compound is a free base, an addition salt (e.g., a pharmaceutically acceptable addition salt) can be prepared by dissolving the free base in a suitable organic solvent and treating the solution with an acid. Conventional methods for the preparation of acid addition salts of active compounds are consistent. Those skilled in the art will recognize various synthetic methods that can be used to prepare non-toxic pharmaceutically acceptable addition salts without undue experimentation.

"Treating", "treat", "treatment" or "mitigation" refers to the administration of at least one compound and / or at least one stereoisomer thereof, and / or at least one medicine Acceptable salts are given to individuals identified as in need thereof, for example, said individuals have cancer.

"Effective amount" means an amount of at least one compound and / or at least one stereoisomer thereof, and / or at least one pharmaceutically acceptable salt in the present invention, which amount is effective to "treat" a disease in an individual Or disease, and to a certain extent, biological, or medical, responses to tissues, systems, animals, or humans, such as one or more symptoms or the disease being treated, are sufficient to prevent it Development, or lightening to some extent. The therapeutically effective amount of the mammal to be treated will vary depending on the compound, the disease and its severity, age, weight, etc.

The term "at least one substituent" as used herein includes, for example, from 1 to 4, such as from 1 to 3, and further, such as from 1 to 2 substituents. For example, "at least one substituent R ¹⁶ " described herein includes from 1 to 4, such as from 1 to 3, and further, such as from 1 to 2 substituents selected from the list of R ¹⁶ described herein.

The present invention provides methods and compositions for inhibiting BRD ₄ and treating diseases caused by poor BRD ₄ activity (BRD ₄ related diseases).

The invention provides a compound of the formula:

Its stereoisomers, and its pharmaceutically acceptable salts.

Each combination of the present invention has been enumerated in detail, including all the combinations of the specific examples listed.

R ¹ is hydrogen, halogen, heteroatom functional group or hydrocarbon group, and the hydrocarbon group is selected from C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, and C6-C14 aryl, among which alkyl, alkenyl The alkynyl system is selectively cyclized, each hydrocarbon group is optionally substituted, and optionally contains 1 to 3 heteroatoms.

In a specific embodiment, R ¹ is a halogen, in particular F, Cl, Br or I.

In a specific embodiment, R ¹ is a heteroatom functional group, the functional group is connected via a heteroatom such as oxygen, nitrogen, phosphorus, or sulfur, and includes a hydroxyl group, an alkoxy group, an aryloxy group, an amine group, and an azo group. , Cyano, thiocyano, peroxy, imine, aldimine, isonitrile, isonitrile, nitrate, nitrile, nitrite, nitro, nitroso, phosphate, phosphine, sulfur Compounds, sulfofluorenyl, sulfo or mercapto, especially R ¹ is -OR ⁴ or -NR ⁵ R ⁶ , wherein R ⁴ , R ⁵ and R ⁶ are each H or a hydrocarbyl group; the hydrocarbyl group is selected from a C1-C8 alkyl group , C2-C8 alkenyl, C2-C8 alkynyl, and C6-C14 aryl; wherein each alkyl, alkenyl, and alkynyl are selectively cyclized, and each hydrocarbyl is selectively substituted, and selected The atom containing 1 to 3 heteroatoms, in which R ⁵ and R ⁶ are attached together, forms a selectively substituted cyclic hydrocarbon. Examples of this ring include optionally substituted 3, 4, 5, 6, 7, and 8-membered rings containing 1, 2, or 3 N heteroatoms and 0, 1, 2, or 3 non-N heteroatoms, including imidazole (Imidazole), pyrazoline (pyrazole), Oxazoline Azole), thiazoline (thiazole), piperidinyl, pyrrolidinyl, piperidine , Morpholine Morpholine, triazole, Diazole, thiadiazole, dithiazole, etc.

In a specific embodiment, R ¹ is a C1-C14 hydrocarbon group, particularly a C1-C8 alkyl group, a C2-C8 alkenyl group, a C2-C8 alkynyl group, or a C6-C14 aryl group, wherein each alkyl group, alkenyl group, The alkynyl group is selectively cyclized, and each hydrocarbyl group is selectively substituted, and optionally contains 1 to 3 heteroatoms. In a specific embodiment, R ¹ is a C3-C8 cycloalkyl group, a C5-C8 cycloalkenyl group, or a C6-C14 aryl group, wherein each hydrocarbon group is optionally substituted and optionally contains 1-3 heteroatoms.

Example R ¹ moiety comprises hydroxyl, amine, phenyl, 3-amino-piperidinyl, 3-hydroxy piperidinyl, hydroxymethyl -, isopropyl - methylamino and the like.

R ² is a heteroatom functional group or a hydrocarbon group, as described for R ¹ .

Exemplary R ² moiety contains cyclohexyl, phenyl, thiophene, phenyl-, hydroxymethyl-methylamino, cyclohexyl-2-one, methyl-, hydroxymethylmethylamino, 2-hydroxymethyl- , 4-hydroxypyrrolidine, 2-carboxy-, 4-hydroxypyrrolidine, hydroxymethyl-, 5-imidazolemethyl-methylamine, 2-hydroxypropylamine, and the like.

R ³ is halogen, optionally substituted, especially halides, especially lower alkyl, hydroxyl, lower alkoxy, or low molecular fluorenyl substituted with F, Cl or Br; examples include F, Cl, Br, CH ₃ , CF ₃ , OCH ₃ and OCOCH ₃ . In specific examples, n is equal to 0, 1, 2 or 3.

The invention provides all compounds, stereoisomers and pharmaceutically acceptable salts in the tables and examples.

In another aspect, the invention provides a substituted 5- (3,5-dimethyliso Azole-4-yl) indoline-2-one BRD ₄ inhibitor, and in the specific examples listed, the inhibitor includes indoline with 1 to 2 substituents at the C3 position, where the The iso-substituents are heteroatom, heteroatom functional group or hydrocarbon group, respectively. The hydrocarbon group is selected from C1-C8 alkyl group, C2-C8 alkenyl group, C2-C8 alkynyl group, and C6-C14 aryl group, among which alkyl group, olefin group Group, alkynyl group is selectively cyclized, each hydrocarbyl group is optionally substituted, and optionally contains 1 to 3 heteroatoms. In a specific embodiment, the substituent is selected from R ¹ , R ² and R ^{3 in} Formula I.

The compound of the present invention, its stereoisomers, and its pharmaceutically acceptable salts can be used alone or in combination with at least one other therapeutic agent. In some specific examples, the present compound, its stereoisomers, and its pharmaceutically acceptable salts can be used in combination with at least one additional therapeutic agent. At least one additional therapeutic agent may be selected from, for example, anti-proliferative, anti-cancer, and chemotherapeutic agents. The compounds disclosed herein and / or a pharmaceutically acceptable salt may be administered in a single dosage form or separate dosage forms with at least one other therapeutic agent. When the separate dosage forms are administered, at least one other therapeutic agent may be administered at the same time, or the compound and / or the pharmaceutically acceptable salt disclosed in the present case may be administered at the same time.

A "chemotherapeutic agent" is a compound used to treat cancer regardless of the mechanism of action. Chemotherapy agents can be compounds used in "targeted therapy" and conventional chemotherapy. Suitable chemotherapeutic agents may be selected from, for example, agents that cause apoptosis; polynucleotides (e.g., ribozymes); polypeptides (e.g., enzymes); drugs; biological mimics; alkaloids; alkylating agents; antitumor Antibiotics; antimetabolites; hormones; platinum compounds; monoclonal antibodies linked to anticancer drugs, toxins, and / or radionuclides; biological response modifiers (interferons, such as IFN-a and interleukins, such as IL -2); immunotherapy agents; hematopoietic growth factors; agents that cause tumor cell differentiation (such as all-trans-vitamin A acid); gene therapy agents; anti-femoral therapy agents and nucleosides; tumor vaccines; and angiogenesis inhibitors .

Examples of chemotherapeutic drugs include Erlotinib (TARCEVA®, Genentech / OSI Pharm.); Bortezomib (VELCADE®, Millennium Pharm.); Fulvestrant (FASLODEX®, AstraZeneca); Shu Nitinib (Sunitinib, SUTENT®, Pfizer); Letrozole (FEMARA®, Novartis); Imatinib mesylate (GLEEVEC®, Novartis); PTK787 / ZK 222584 (Novartis); Austria Oxaliplatin, Eloxatin®, Sanofi; 5-FU (5-fluorouracil); Leucovorin; Rapamycin (Sirolimus, RAPAMUNE®, Wyeth); La Patinib (Lapatinib, TYKERB®, GSK572016, GlaxoSmithKline); Lonafarnib (SCH 66336); Sorafenib (NEXAVAR®, Bayer); Irinotecan (CAMPTOSAR®, Pfizer) and gefitinib (Gefitinib, IRESSA®, AstraZeneca); AG1478, AG1571 (SU 5271, Sugen); alkylating agents such as thiotepa and CYTOXAN® cyclophosphamide; alkylsulfonic acid Salts, such as busulfan, improsulfan, and piperbutane (piposulfan); aziridines, such as benzodopa, carboquone, meturedopa, and uredopa; ethyleneimine and methylmelamine (methylamelamines) such as hexamethylmelamine, triethyl melamine, triethyl ethylphosphonium amine, triethyl ethyl phosphorothioate, and trimethyl melamine; lactones (such as bulatacin) And bulatacinone); camptothecin (such as a synthetic analog of topotecan); bryostatin; callystatin; CC-1065 and its adolesine (adozelesin), carzelesin, synthetic analogs of bizelesin; cryptophycins (eg, candida 1 and candida 8); dolastatin; Duocarmycin and its synthetic analogs, such as KW-2189 and CB1-TM1; eleutherobin; pancratistatin; sarcodictyin; spongistatin ); Nitrogen mustard, such as chlorambucil, chlomaphazine, chlorophosphamide, estrogen mustard, ifosfamide famide), mechlorethamine, methanothionine hydrochloride, melphalan, novembichin, benzyl cholesterol, phenesterine, prednimustine, trisalin Trofosfamide, uracil nitrogen mustard; nitrosourea such as carmustine, chlorozotocin, fotemustine, lomustine, nepal Mustin (nimustine) and ranimnustine; antibiotics such as enediyne antibiotics (such as calicheamicin, such as calicheamicin γ1I and calicheamicin ωI1 (Angew Chem .Intl.Ed.Engl. (1994) 33: 183-186)); dynemicin, such as dynemicin A; bisphosphonates, such as clodronate; (esperamicin); and neocarzinostatin chromophore, and related chromophore chromophores, aclacinomysins, actinomycin, anqu Autramycin, azaserine, bleomycin, cactinomycin, carabici n), caminomycin, carzinophilin, chromomycinis, actinomycin D (dactinomycin), daunorubicin; detorubicin, 6 -Diazo-5-oxo-L-n-leucine, ADRIAMYCIN® (doxorubicin), Porphyrin-doxorubicin, cyano Porphyrin-doxorubicin, 2-pyrrolidinyl-doxorubicin and deoxorubicin, epirubicin, esorubicin, idarubicin, hemp Marcellomycin, mitomycin such as mitomycin C, mycophenolic acid, nogalamycin, olivomycins, peplomycin, pofomycin (porfiromycin), puromycin, quelamycin, rodorubicin, streptozotocin, streptozotocin, tubercidin, urbenes (ubenimex), netinostatin (zinostatin), zorubicin; antimetabolites such as methamphetamine and 5-fluorouracil (5-FU); folic acid analogs such as diopterin, amine Methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as Cyclocytidine (ancitabine), azacitidine, 6-azauridine, carmofur, cytarabine, dideoxy Glycosides, deoxyfluridine, enocitabine, fluorouridine; androgens such as calusterone, drostmostanolone propionate, epitiostanol , Mepitiostane, galactone; anti-adrenalin such as aminoglutethimide, mitotane, trilostane; folic acid supplements such as frolinic acid; vinegar and glucose Aceglatone; aldothionine; aminol evulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene ); Edatraxate; Defofamine; Demecolcine; Diaziquone; Elephithine; Elliptinium acetate; Epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids, such as maytansine ) And ansamitocins; mitoguazone; mitoxanthin mitoxantrone; mopidanmol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone Podophyllinic acid; 2-ethylhydrazine; procarbazine; PSK® polysaccharide complex (JHS Natural Products, Eugene, Oreg.); Razoxane; radixomycin ( rhizoxin); sizofuran; spirogermanium; tenuazonic acid; triaziquone; 2,2'2 "-trichlorotriethylamine; Trichothecenes (such as T-2 toxin, verracurin A, roridin A, and anguidine); urthan; vindesine (vindesine); dacarbazine; mannitol nitrogen mustard; dibromomannitol (mitobronitol); dibromo dulcitol (mitolactol); pipobroman; gacytosine; ("Ara-C");cyclophosphamide;thiotepa; taxanes, for example, TAXOL® (paclitaxel; Bristol-Myers Squibb O ncology, Princeton, NJ), ABRAXANE® (Cremophor-free), albumin engineered paclitaxel nanoparticle formulations (American Pharmaceutical Partners, Schaumberg, Ill.), and TAXOTERE® (European paclitaxel ( doxetaxel); Rhone-Poulenc Rorer, Antony, France); chloranmbucil; GEMZAR® (gemcitabine); 6-thioguanine; mercaptopurine; methotrexate; platinum analogs, Such as cisplatin, carboplatin; vinblastine; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; NAVELBINE® (vinca Vinorelbine); mitantrone; teniposide; edatrexate; daunomycin; aminopterin; capecitabine XELODA®); ibandronate; CPT-11; topoisomerase inhibitor RFS 2000; difluoromethyl ornithine (DMFO); retinoids such as retinoic acid; and any of the above Pharmaceutically acceptable salts, acids and derivatives.

"Chemotherapeutic agents" may also be selected from, for example: (i) antihormonal agents capable of regulating or inhibiting the action of hormones on tumors, such as antiestrogens and selective estrogen receptor modulators (SERMs), including, for example, it Tamoxifen (including NOLVADEX®; tamoxifen citrate), raloxifene, droloxifene, 4-hydroxytamoxifen, trioxifene , Raloxifene hydrochloride (keoxifene), LY117018, onapristone and FARESTON® (toremifene citrate (toremifine citrate)); (ii) aromatase inhibitors capable of inhibiting aromatase, which can regulate the production of estrogen in the adrenal glands, such as 4 (5) -imidazole, aminoglutethimide, MEGASE® Progesterone acetate), AROMASIN® (exemestane; Pfizer), Formestanie, fadrozole, RIVISOR® (vorozole), FEMARA® (letrozole) (letrozole); Novartis) and ARIMIDEX® (anastrozole; AstraZeneca); (iii) antiandrogens such as flutamide, nilitamide, bicalutamide, Leuprolide and goserelin; and troxacitabine (1,3-dioxetane nucleoside cytosine analog); (iv) protein kinase inhibitors; (iv) v) Lipid kinase inhibitors; (vi) Anti-strand oligonucleotides, which can inhibit the expression of genes in the signal transduction pathway, these genes cause the appreciation of some mutant cells, such as PKC-α, Ralf and H-Ras; (vii ) Ribozymes, such as VEGF performance inhibitors (such as ANGIOZYME®) and HER2 performance inhibitors; (viii) vaccines, such as gene therapy vaccines, such as ALLOVECTIN® , LEUVECTIN®, and VAXID®; PROLEUKIN® rIL-2; topoisomerase I inhibitors such as LURTOTECAN®; ABARELIX® rmRH; (ix) anti-angiogenic agents such as bevacizumab (AVASTIN®, Genentech); and (x) any of the aforementioned pharmaceutically acceptable salts, acids, and the like.

The "chemotherapeutic agent" may also be selected from antibodies with curative effects such as alemtuzumab (Campath), bevacizumab (AVASTIN®, Genentech); cetuzimab (ERBITUX®, Imlone) ); Panitumumab (VECTIBIX®, Amgen), rituximab (RITUXAN®, Genentech / Biogen Idec), pertuzumab (OMNITARG®, 2C4, Genentech), trastuzumab Beadizumab (tratuzumab, HERCEPTIN®, Genentech), tositumomab (toxitumomab, becoxacin) (Bexxar), Corixia), and an antibody drug conjugate, gemtuzumab ozogamicin (MYLOTARG®, Wyeth).

Potentially effective humanized monoclonal antibodies can be used as chemotherapeutic agents to combine the compounds of this case and their isomers, as well as pharmaceutically acceptable salts, such as selected from: alenzumab, apolizumab , Aselizumab, atlizumab, bapineuzumab, bevacizumab, bivatuzumab mertansine, mocantozumab ( cantuzumab mertansine), cedelizumab, certolizumab pegol, cidfusituzumab, cidtuzumab, daclizumab, Eculizumab, efalizumab, epratuzumab, erlizumab, felvizumab, fangtuuzumab (fontolizumab), gemtuzumab, ommtuzumab ozogamicin, inotuzumab ozogamicin, ipilimumab, labetuzumab, lin Toltuzumab, matuzumab, mepolizumab ), Motavizumab, motovizumab, natalizumab, nimotuzumab, nolovizumab, Numavizumab, ocrelizumab, omalizumab, palivizumab, pascolizumab, pefcilizumab (pecfusituzumab), pectuzumab, pertuzumab, pexelizumab, ralivizumab, ranibizumab, ruilizumab Resilivizumab, resilizumab, resyvizumab, rovelizumab, ruplizumab, sirolimumab (sibrotuzumab), siplizumab, soltuzumab, tacatuzumab tetraxetan, tadocizumab, talizumab , Tefibazumab, tocilizumab, toralizumab, trastuzumab, tucotuzumab celmoleukin, toctozumab Tucusituzumab, umavizumab, urtoxazumab, and visilizumab.

Also provided is a composition comprising the present compound, an isomer thereof, and a pharmaceutically acceptable salt, and at least one pharmaceutically acceptable carrier.

The composition contains the present compound and its isomers, and a pharmaceutically acceptable salt thereof, and can be administered by various known methods, such as oral, topical, rectal, parenteral, inhalation spray, Or via implantable reservoirs, although the most suitable route of administration under certain assumed conditions depends on the particular host, the natural conditions and the severity of the disease when the active ingredient is administered. The term "parenteral administration" includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intraarterial, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques. The compositions disclosed herein may conveniently be in unit dosage form and prepared by any method known in the art.

The compounds of the present invention and its isomers, and their pharmaceutically acceptable salts, can be administered orally in solid dosage forms, such as capsules, dragees, tablets, dragees, granules and powders, or in liquid dosage form, such as tincture Agents, syrups, emulsions, dispersions and suspensions. The compound and its isomers, and pharmaceutically acceptable salts thereof, can also be administered parenterally in sterile liquid dosage forms, such as dispersions, suspensions or solutions. The compound and its isomers and their pharmaceutically acceptable salts can also be administered in other dosage forms, such as ointments, creams, drops, topical transdermal patches or powders, and ophthalmic solutions. Or hanging Liquids such as eye drops, ophthalmic medication, inhalation or nasal administration in the form of a spray or powder composition, or rectal or vaginal administration in the form of creams, ointments, sprays, suppositories.

Gelatin capsules contain the present compound and / or at least one pharmaceutically acceptable salt and a powdery carrier such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid and the like can also be used. Similar diluents can be used to make compressed lozenges. Both tablets and capsules can be made into sustained release products for continuous administration over a period of time. Compressed tablets can be coated with sugar or film to mask the unpleasant taste while protecting the tablets from the atmosphere, or enteric coatings can be selectively dissolved in the gastrointestinal tract.

The liquid dosage form for oral administration further includes at least one selected from the group consisting of colorants and flavoring agents for improving patient acceptance.

Generally, water, suitable oils, saline solutions, dextrose in water (glucose), and related sugar solutions and glycols such as propylene glycol or polyethylene glycol can be suitable carriers for parenteral administration solutions. The parenteral solution contains a water-soluble salt of at least one compound described herein, at least one suitable stabilizer, and if necessary at least one buffer. Some antioxidants such as sodium bisulfite, sodium sulfite, or ascorbic acid, alone or in combination, can be suitable stabilizers. Citric acid and its salts and sodium ethylenediamine tetraacetate may also be suitable stabilizers. In addition, the parenteral administration solution may further include at least one preservative selected from, for example, benzalconium chloride, methyl- and propylparaben and chlorobutanol.

Pharmaceutically acceptable carriers can be selected from carriers that are compatible with the active ingredients in the composition (in some specific cases, they can stabilize the active ingredients) and that are not harmful to the individual being treated. Solubilizers, such as cyclodextrin (which can form specific, more soluble complexes with the disclosed at least one compound and / or at least one pharmaceutically acceptable salt), are used as pharmaceutical excipients for transport Active ingredient. Examples of other carriers include colloidal silica, magnesium stearate, cellulose, sodium lauryl sulfate, and pigments such as D & C Yellow No. 10. Suitable pharmaceutically acceptable carriers are described in Remington's Pharmaceutical Sciences , A. Osol, and can be used as standard text in the art.

The compound and its isomers, and their pharmaceutically acceptable salts can be further used to test the efficacy of BRD ₄ related diseases in vivo. For example, the present compound and / or at least one pharmaceutically acceptable salt thereof can be administered to an animal (such as a mouse model) suffering from a disease associated with BRD ₄ to evaluate its therapeutic effect. A positive result in one or more of these tests can effectively enhance the scientific knowledge base and thus effectively show the utility of these compounds and / or salts. Based on these conclusions, an appropriate dose range and route of administration for animals such as humans can be determined.

For inhaled administration, the compound and its isomers, as well as their pharmaceutically acceptable salts, can be conveniently delivered from a compressor or sprayer in the form of an aerosol spray. The compound and its isomers, as well as the pharmaceutically acceptable salts thereof, can also be formulated into an inhaled powder composition for transmission with the aid of an inhalation dry powder inhaler device. A metered-dose inhalation nebulizer (MDI) is an exemplary inhalation delivery system that can combine the compounds disclosed herein and their isomers, and their pharmaceutically acceptable salts in at least one suitable propellant such as selected from fluorocarbons and hydrocarbons The compounds are formulated into suspensions or solutions.

For ophthalmic administration, ophthalmic preparations can be prepared by mixing a suitable amount of a solution or suspension of the compound and its isomers with a pharmaceutically acceptable salt in a suitable ophthalmic carrier, such that The compound and its isomer, and at least one pharmaceutically acceptable salt thereof, are kept in contact with the surface of the eyeball for a sufficient time to allow the compound to penetrate into the cornea and internal regions of the eye.

The compounds and their isomers disclosed herein, and their pharmaceutically acceptable salts, useful pharmaceutical dosage forms for administration include, but are not limited to, hard and soft gelatin capsules, dragees, parenteral injections, and oral suspensions.

The dosage is related to many factors, such as age, the health and weight of the subject, the extent of the disease, the type of treatment being performed, if any, the frequency of treatment and the desired effect. In general, the daily dosage of the active ingredient can be varied, such as from 0.1 to 2000 mg per day. For example, 10-500 mg once or more a day may achieve the desired effect.

In some specific examples, a large number of unit capsules can be prepared by filling two standard hard gelatin capsules each, for example, 100 mg of the compound and its isomer disclosed herein in powder form, and a pharmaceutically acceptable salt thereof. , 150 mg lactose, 50 mg cellulose, and 6 mg magnesium stearate.

In some specific examples, the compound, its isomer, and its pharmaceutically acceptable salt and digestible oil, such as a mixture of soybean oil, cottonseed oil, or olive oil, can actively replace the way of pumping gelatin Prepare or inject to form soft gelatin capsules containing 100 mg of active ingredient. These capsules are washed and dried.

In some specific examples, a large number of lozenges can be prepared by conventional procedures, and each dosage unit contains, for example, 100 mg selected from the compound and its isomers, and a pharmaceutically acceptable salt thereof, 0.2 mg of colloidal Silica, 5 mg magnesium stearate, 275 mg microcrystalline cellulose, 11 mg starch and 98.8 mg lactose. A suitable coating can be used to improve palatability or delay absorption.

In some specific examples, the parenteral composition suitable for injection administration can be stirred at 1.5% by weight of the disclosed compound and / or at least one enantiomer, diastereomers The structure, or a pharmaceutically acceptable salt thereof, is prepared in 10% by volume propylene glycol. This solution was added with water for injection to the desired volume and sterilized.

In some embodiments, an aqueous suspension for oral administration can be prepared. For example, each 5 ml of aqueous suspension contains 100 mg of finely divided compounds, their stereoisomers, and their pharmaceutically acceptable salts, 100 mg of sodium carboxymethyl cellulose, 5 mg of sodium benzoate, 1.0 g Sorbitol solution, USP and 0.025 ml vanillin.

When the present compound, its isomer, and its pharmaceutically acceptable salt are administered stepwise or together with at least one other therapeutic agent, the same dosage form can be used. When drugs are administered in physical combinations, the choice of dosage form and route of administration depends on the compatibility of the combination drugs. The term simultaneous administration can therefore be understood as meaning that at least two agents are administered simultaneously or sequentially, or are administered in a fixed dose of a composition of at least two active ingredients.

The compounds disclosed in this case, their isomers, and their pharmaceutically acceptable salts can be administered as a single active ingredient or with at least one second active ingredient, such as those selected from other known agents for treating patients with BRD ₄ related diseases The active ingredient is administered in combination.

It can be understood that the embodiments and specific examples described herein are only for the purpose of enumeration. Those skilled in the art can make various modifications or changes accordingly, and these modifications and changes are included in the spirit and scope of the present application. Within the scope of the attached patent application. The entire contents of all publications, patents, and patent applications cited herein are incorporated herein by reference for all purposes.

General formula for compound preparation

The compounds of the present invention, their stereoisomers and pharmaceutically acceptable salts can be prepared from (a) commercially available starting materials; (b) known starting materials prepared according to procedures described in the literature (c) according to the scheme and Experimental procedures describe new intermediates. In synthesizing the compound of the present invention, in order to improve the yield of the compound required for synthesis, the order of the synthetic steps may be changed. Turn into Compounds and / or pharmaceutically acceptable salts can be synthesized from commercially available materials and materials prepared by the present invention. The following reaction schemes describe some methods for preparing the disclosed compounds.

In this reaction formula, a commercially available 5-bromoindole-2,3-dione 1 is converted into a ketal intermediate 2 and then subjected to a Suzuki coupling reaction with boric acid 3. The resulting ketal-protected intermediate 4 is Deprotection gives 5- (3,5-dimethyliso Azole-4-yl) indolin-2,3-dione. Under standard conditions known in the art, this key intermediate 5 and an organometallic reagent (such as a Grignard reagent) undergo an addition reaction to form a compound of formula Ia .

Scheme II describes some methods for preparing the compounds ( Ib ) and ( Ic ) disclosed herein .

In this reaction formula, under standard conditions known in the art, the compound Ia is treated with thallium chloride to give a compound of formula Ib , and then the chloro group is substituted with an amine or a hydroxyl group to give a compound of formula Ic .

Scheme III describes some methods for the preparation of compounds ( Id ) and ( Ie ) disclosed herein .

In this reaction formula, under standard conditions known in the art, compounds Ia of the hydroxyl group with an acidic condition triethyl silane-lower (e.g., trifluoroacetic acid) is reduced to a compound of formula Id, a compound of Formula Id by alkylation A compound of formula Ie is obtained.

Scheme IV describes some methods for preparing the disclosed compounds ( If ).

In this reaction scheme, under standard conditions known in the art, (R) -2-methylpropane-2-sulfimidine is introduced by 5- (3,5-dimethyliso The azole-4-yl) indol-2,3-dione produces compounds of formula 6 under Lewis acid conditions and is then protected by amine groups. Under known standard conditions, the addition reaction of key intermediate 7 and an organometallic reagent (such as a format reagent) gives a compound of formula 8 , which is deprotected under acidic conditions known in the art to give a compound of formula If .

Scheme V describes some methods for preparing the disclosed compound ( Ig) .

Reaction Formula V

In this reaction formula, under standard amidine coupling conditions known in the art, the resulting amine and acid coupling of formula 8 yields a compound of formula Ig .

Examples

The purpose of the following examples is purely exemplary and should not be limited in any way. Although every effort has been made to ensure the accuracy of the information (eg, volume, temperature, etc.), some experimental errors and deviations are inevitable. Unless stated otherwise, temperatures are in Celsius. Reagents were purchased from commercial suppliers such as Sigma-Aldrich, Alfa Aesar, or TCI, and were used directly without further purification unless otherwise specified.

Unless otherwise stated, the following reactions are performed in anhydrous solvents, under the positive pressure of nitrogen or argon or using a drying tube; the reaction bottle is equipped with a rubber septum, and the matrix and reagents are added through a syringe; glassware is dried and / or heated to dry .

Unless otherwise stated, the following reactions are performed in anhydrous solvents, under the positive pressure of nitrogen or argon or using a drying tube; the reaction bottle is equipped with a rubber septum, and the matrix and reagents are added through a syringe; glassware is dried and / or heated to dry .

Unless otherwise noted, column chromatography was performed on a Biotage system with a silica gel column (manufacturer: Dyax), or using a silica silica SepPak cartridge (Waters), or a pre-packed silica gel column on a Teledyne Isco Combiflash purification system.

¹ H NMR spectra were recorded by a Varian device operating at 400 MHz. The obtained ¹ H-NMR spectrum was obtained using CDCl ₃ , CD ₂ Cl ₂ , CD ₃ OD, D ₂ O, d ₆ -DMSO, d ₆ -acetone or (CD ₃ ) ₂ CO as a solvent, and tetramethylsilane ( 0.00ppm) or based on residual solvent (CDCl ₃ : 7.25 ppm; CD ₃ OD: 3.31 ppm; D ₂ O: 4.79 ppm; d ₆ -DMSO: 2.50 ppm; d ₆ -acetone: 2.05; (CD ₃ ) ₂ CO: 2.05). When indicating peak shape diversity, the following abbreviations indicate different peak shapes: s (single peak), d (double peak), t (triple peak), q (quadruplex), qn (quintet), and sx (six Heavy peak), m (multiple peak), br (broad peak), dd (two double peaks), dt (two triplet peaks). Coupling constants are in Hertz (Hz). Compound names were generated with the exception of reagents by ChemDraw version 12.0.

abbreviation:

Example 1: Synthesis of compounds 1.1 to 1.34

Compound 1.1

Step 1: 5'-Bromospiro [[1,3] dioxolane-2,3'-dihydroindole] -2'-one

5-Bromodihydroindole-2,3-dione (226g, 1.0mol, 1eq.), Ethylene glycol (186g, 3.0mol, 3eq.) And p-TSA (30g, 0.16mol, 10mol%) were dissolved. In toluene (1500 ml), reflux using a Dean-Stark apparatus overnight. After all 5-bromoindoline-2,3-dione was reacted, EtOAc (1500 mL) and water (1000 mL) were added to remove excess diol. The aqueous phase was extracted twice with EtOAc (500 mL), and the organic phases were combined. Dry over sodium sulfate, filter, and concentrate under reduced pressure to give a yellow solid, then wash with ether to the desired compound as a pale yellow solid (236 g, 87%). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ _H 10.56 (s, 1H), 7.31-7.33 (m, 2H), 6.93 (d, J = 8.0 Hz, 1H), 4.27-4.35 (m, 4H) , 2.35 (s, 3H), 2.18 (s, 3H). MS (ESI) m / e [M + 1] ⁺ 270,272.

Step 2: 5 '-(3,5-dimethyliso Azole-4-yl) spiro [[1,3] dioxolane-2,3'-dihydroindole] -2'-one

3,5-dimethyliso Azole-4-boronic acid (159g, 1.13mol), dichloro [1,1'-bis (di-third-butylphosphino)] ferrocene] palladium (II) (23.6g, 29mmol), 5'-bromo Spiro [[1,3] dioxolane-2,3'-dihydroindole] -2'-one (236g, 0.87mol) and sodium carbonate (184g, 1.7mol) are dissolved in dioxane / water The mixed solution (1200 mL / 300 mL) was heated to 110 ° C for 12 hours. The mixture was filtered through a celite pad and concentrated under reduced pressure. The crude product was dissolved in dichloromethane (2L), then n-hexane (2L) was added, and filtered again. The organic phase was concentrated under reduced pressure to obtain the target product (150 g, 60%). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ _H 10.56 (s, 1H), 7.31-7.33 (m, 2H), 6.93 (d, J = 8.0 Hz, 1H), 4.27-4.35 (m, 4H) , 2.35 (s, 3H), 2.18 (s, 3H). MS (ESI) m / e [M + 1] ⁺ 287.

Step 3: 5- (3,5-Dimethyliso Azole-4-yl) indolin-2,3-dione

5 '-(3,5-dimethyliso Azole-4-yl) spiro [[1,3] dioxolane-2,3'-dihydroindole] -2'-one (150g, 0.52mol), acetic acid (10ml) and concentrated hydrochloric acid solution ( (300 ml) was heated to 90 ° C. After 1 hour, the mixture was poured into ice water, filtered and washed with water, ethanol and ethyl acetate to collect the desired compound 5- (3,5-dimethyliso Azole-4-yl) indoline-2,3-dione (115 g, 92%). ¹ H NMR (DMSO- d ₆ ) δ _H 11.14 (s, 1H), 7.59 (d, J = 8.0Hz, 1H), 7.51 (s, 1H), 7.00 (d, J = 8.0Hz, 1H), 2.37 (s, 3H), 2.20 (s, 3H) .MS (ESI) m / e [M + 1] ⁺ 243.

Step 4: 5- (3,5-Dimethyliso Azole-4-yl) -3-hydroxy-3-phenylindol-2-dione

5- (3,5-dimethyliso Azole-4-yl) indolin-2,3-dione (24.2 g, 0.1 mol) was dissolved in anhydrous THF (400 mL) and cooled to 0 ° C, and then a 2.0 M solution of PhMgBr in THF ( 110mL, 220.0mmol), the ice bath was removed, the reaction was stirred under nitrogen for 30 minutes, and the complete consumption of raw materials was detected by TLC. The reaction mixture was quenched by the addition of a saturated aqueous NH ₄ Cl solution (100 mL), and extracted with ethyl acetate (3 × 100 ml). The organic phases were combined, washed with saturated aqueous sodium bicarbonate (100 mL) and brine (100 mL), dried over anhydrous magnesium sulfate, filtered, concentrated under reduced pressure, and degassed under high vacuum to obtain the crude alcohol (30 g, 94%). ¹ H NMR (400MHz, DMSO- d ₆ ): δ 10.50 (s, 1H), 7.22-7.32 (m, 6H), 7.09 (d, J = 1.2Hz, 1H), 6.99 (d, J = 8.0Hz, 1H), 6.71 (brs, 1H), 2.32 (s, 3H), 2.15 (s, 3H). MS (ESI) m / e [M + 1] ⁺ 321.

Compounds 1.1a and 1.1b

Each racemic 1.1a and 1.1b enantiomeric system was separated by preparative HPLC with Chiralpak AD using 25% 2-propanol / hexane as the eluent. The enantiomeric excess was determined by HPLC of Chiralpak AD, using 25% 2-propanol / hexane as the eluent, and the flow rate was 1.0 mL / min. The retention time of the first enantiomer after elution was 5.8 minutes The retention time of the other enantiomer after elution was 7.4 minutes. The title compound has exactly the same spectral properties as 1.

The following compounds 1.2 to 1.29 were synthesized using similar procedures described in compound 1.1 starting from the corresponding format reagents.

Compounds 1.30a and 1.30b

5- (3,5-dimethyliso Azol-4-yl) -3-hydroxy-3- (2-oxocyclohexyl) indol-2-one

At room temperature, compound 5- (3,5-dimethyliso Azole-4-yl) indolin-2,3-dione (242 mg, 1.0 mmol) was dissolved in methanol (20 ml), cyclohexanone (196 mg, 2.0 mmol) and dimethylamine (0.4 mmol) were added, The reaction mixture was stirred for 4 h. Thereafter, the mixture was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain two isomers (the first 50 mg, 14.7%, the second 30 mg, 8.8%, and the mixture 100 mg, 29.3%). Fast-acting isomer (50mg, 14.7%): ¹ H NMR (400MHz, DMSO- d ₆ ): δ 10.25 (s, 1H), 7.29 (d, J = 1.6Hz, 1H), 7.17 (dd, J = 1.6, 8.0Hz, 1H), 6.84 (d, J = 8.0Hz, 1H), 5.83 (s, 1H), 3.32-3.41 (m, 1H), 2.30-2.40 (m, 5H), 2.19 (s, 3H), 1.98-2.06 (m, 2H), 1.80-1.83 (m, 2H), 1.44-1.64 (m, 2H), MS (ESI) m / e [M + 1] ⁺ 341.0; slow reaction isomerization (30mg, 8.8%): ¹ H NMR (400MHz, DMSO- d ₆ ): δ 10.31 (s, 1H), 7.16-7.20 (m, 2H), 6.88 (d, J = 8.0Hz, 1H), 5.92 (s, 1H), 3.08-3.12 (m, 1H), 2.59-2.62 (m, 1H), 2.36 (s, 3H), 2.29-2.35 (m, 1H), 2.19 (s, 3H), 1.77-2.00 (m, 4H), 1.62-1.68 (m, 1H), 1.42-1.50 (m, 1H), MS (ESI) m / e [M + 1] ⁺ 341.

3-chloro-5- (3,5-dimethyliso Azole-4-yl) -3-phenylindol-2-one

5- (3,5-dimethylisopropyl) at 0 ℃ Azol-4-yl) -3-hydroxy-3-phenylindol-2-one (30 g, 0.094 mol) in THF (300 ml) was added with pyridine (40 ml), and then chlorinated Asiatic (20 ml). The reaction mixture was stirred at 0 ° C for 0.5 h, and quenched by adding saturated ammonium chloride solution (50 mL). The organic phase was extracted with saturated ammonium chloride (2 × 50 ml). The combined aqueous phases were extracted with dichloromethane (200 mL). The combined organic phases were dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to give the target compound (25g, 79 %). ¹ H NMR (400MHz, DMSO-d ₆ ): δ 11.11 (s, 1H), 7.37-7.53 (m, 6H), 7.08 (d, J = 8.0Hz, 1H), 2.30 (s, 3H), 2.20 ( s, 3H).

5- (3,5-dimethyliso Azole-4-yl) -3- (2-hydroxyethoxy) -3-phenylindol-2-one

30 mg (0.088 mmol) of 3-chloro-5- (3,5-dimethyliso Azol-4-yl) -3-phenylindol-2-one and 139 mg (0.176 mmol) of pyridine were dissolved in 5 ml of THF, and 54 mg (0.88 mmol) of ethylene-1,2-diol was added. The reaction solution was stirred at room temperature for 5 hours. Next, the solution was diluted with water, and the aqueous phase was extracted with 2 × 25 mL of ethyl acetate. The combined organic phases were washed with aqueous sodium bicarbonate solution and water, dried and concentrated under reduced pressure. The resulting residue was purified by Pre-TLC (eluent: hexane Hexane / ethyl acetate = 1/1) to give the product (5 mg, 15%) as a white solid. ¹ H NMR (400MHz, DMSO-d ₆ ): δ 10.77 (s, 1H), 7.32-7.35 (m, 6H), 7.20 (s, 1H), 7.04 (d, J = 8.0Hz, 1H), 4.65 ( t, J = 5.6Hz, 1H), 3.23-3.52 (m, 4H), 2.35 (s, 3H), 2.18 (s, 3H). MS (ESI) m / e [M + 1] ⁺ 365.

3-cyclohexyl-5- (3,5-dimethyliso Azole-4-yl) -3- (2-hydroxyethoxy) indol-2-one

Compound 3-cyclohexyl-5- (3,5-dimethyliso Zol-4-yl) -3-hydroxyindolin-2-one was synthesized according to the procedure of Compound 1.32 . ¹ H NMR (400MHz, DMSO-d6): δ 10.59 (s, 1H), 7.27 (d, J = 8.0Hz, 1H), 7.21 (s, 1H), 6.92 (d, J = 8.0Hz, 1H), 4.49 (t, J = 5.6Hz, 1H), 3.37-3.43 (m, 2H), 2.93-3.13 (m, 2H), 2.40 (s, 3H), 2.22 (s, 3H), 1.82-1.90 (m, 2H), 1.56-1.70 (m, 4H), 0.94-1, 15 (m, 4H), 0.74-0.78 (m, 1H). MS (ESI) m / e [M + 1] ⁺ 365.

5- (3,5-dimethyliso Azol-4-yl) -2-oxo-3-phenylindol-3-yl acetate

Compound 3-chloro-5- (3,5-dimethyliso Azol-4-yl) -3-phenylindol-2-one (50 mg, 0.15 mmol) in a solution of dioxane (2 mL) was added with sodium acetate (90 mg, 1.1 mmol), and the reaction mixture was stirred at 85 ° C. Stir at ℃ for 6h, quench with saturated ammonium chloride solution (10mL), extract the organic phase with dichloromethane (2 × 20mL), combine the organic phases to dry over anhydrous sodium sulfate and filter. The filtrate is concentrated under reduced pressure to produce a crude product. It was purified by Pre-TLC (eluent: hexane / ethyl acetate = 1/1) to obtain the product (20 mg, 37%) as a white solid. ¹ H NMR (400MHz, DMSO- d ₆ ): δ 10.78 (s, 1H), 7.33-7.38 (m, 6H), 7.25 (s, 1H), 7.01 (d, J = 8.0Hz, 1H), 2.35 ( s, 3H), 2.18 (s, 3H), 2.15 (s, 3H). MS (ESI) m / e [M + 1] ⁺ 363.

Example 2: Synthesis of compounds 2.1-2.58

5- (3,5-dimethyliso Azole-4-yl) -3- (2-hydroxyethylamino) -3-phenylindol-2-one

34 mg (0.1 mmol) of 3-chloro-5- (3,5-dimethyliso Zol-4-yl) -3-phenylindolin-2-one and 86 mg (10 mmol) of 2-aminoethanol were dissolved in THF (5 mL), and the reaction solution was stirred at room temperature for 16 hours. Then, the solution was diluted with water, and the aqueous phase was extracted with dichloromethane (2 × 10 mL). The combined organic phases were washed with an aqueous sodium hydrogen carbonate solution and water, dried and concentrated under reduced pressure. The obtained residue was subjected to silica gel chromatography (eluent : EtOAc / MeOH = 50/1) purification to give the product (20 mg, 55%) as a white solid. ¹ H NMR (400MHz, DMSO-d ₆ ): δ 10.67 (s, 1H), 7.44 (d, J = 8.0Hz, 1H), 7.18-7.34 (m, 5H), 7.18 (s, 1H), 6.99 ( d, J = 8.0Hz, 1H), 4.51 (t, J = 5.6Hz, 1H), 3.40-3.42 (m, 2H), 2.96-2.99 (m, 1H), 2.40-2.42 (m, 1H), 2.35 (s, 3H), 2.27-2.29 (m, 1H), 2.17 (s, 3H) .MS (ESI) m / e [M + 1] ⁺ 364.

Compound 2.2

Step 1: 5-Bromo-3-hydroxy-3-phenylindol-2-one

At 0 ° C, 5-bromoindoline-2,3-dione (94 g, 416 mmol, 1.0 eq) in THF (1 L), and phenyl magnesium chloride in THF (2.0 M, 500 mL, 1.0 mol) was added dropwise. , 2.4 eq) solution, and the reaction mixture was stirred at room temperature for 2 hours. The mixture was then quenched with saturated aqueous ammonium chloride solution, washed with brine, dried over sodium sulfate, and the solvent was removed to give the crude product. The crude product was then stirred in PE / EA for 1 hour, and the solid was collected by filtration and dried in air to obtain 77 g (61% yield) of the product. ¹ H NMR (400MHz, DMSO- d ₆ ): δ _H 10.57 (s, 1H), 7.42-7.45 (dd, 1H, J = 8, 4, 2.0 Hz), 7.26-7.36 (m, 5H), 7.20- 7.21 (d, 1H, J = 2.0Hz), 6.87-6.89 (d, 1H, J = 8.0Hz), 6.78 (s, 1H).

Step 2: 5- (3,5-Dimethyliso Azole-4-yl) -3-hydroxy-3-phenylindol-2-one

Under nitrogen, remove 3,5-dimethylisopropyl Azole-4-ylboronic acid (28g, 200mmol, 1.5eq), dichloro [1,1'-bis (di-third-butylphosphino) ferrocene] palladium (II) (4.5g, 6.1mmol, 0.05 eq), 5-bromo-3-hydroxy-3-phenylindol-2-one (40 g, 132 mmol, 1.0 eq) and sodium carbonate (45 g, 425 mmol, 3.2 eq) in dioxane / water ( 500 mL / 120 mL) was heated under reflux for 5 hours. After cooling, 500 ml of EA was added, and the mixture was filtered through a pad of celite, and the filtrate was washed with brine (500 mL × 2), dried over sodium sulfate, concentrated, and purified by silica gel to obtain 27.7 g of the product (yield 65.6%). ¹ H NMR (400MHz, DMSO- d ₆ ): δ _H 10.51 (s, 1H), 7.25-7.34 (m, 6H), 7.10 (m, 1H), 6.99-7.01 (d, 1H, J = 8.0Hz) , 6.72 (s, 1H), 2.33 (s, 3H), 2.16 (s, 3H) .MS (ESI) m / e [M + 1] ⁺ 321.

Step 3: 3-Amino-5- (3,5-dimethyliso Azole-4-yl) -3-phenylindol-2-one

Under nitrogen, remove 5- (3,5-dimethyliso A solution of azole-4-yl) -3-hydroxy-3-phenylindolin-2-one (21g, 65.6mmol, 1.0eq) in THF (300ml), drip at -15 ° C ~ -20 ° C Add pyridine (30 ml) and thallium chloride (15 ml), stir the mixture at -15 ° C to -20 ° C for 20 minutes, quench with brine (50 ml), and wash with brine (300 ml x 2). The organic phase was concentrated to give 17.5 g of crude product. The crude product was dissolved in NH ₃ / MeOH (7M, 200 mL) and stirred at room temperature for 2 hours. The solvent was then removed under reduced pressure to obtain the crude product. The crude product was dissolved in EA (200 mL), washed with water (100 mL), then brine (100 mL), dried over sodium sulfate, and concentrated to give 15 g of crude product, which was obtained in silica gel. Purification gave 8.5 g of the product (yield 40%) as a white solid. ¹ H NMR (DMSO- d ₆ ) δ _H 10.53 (s, 1H), 7.39-7.41 (d, 2H, J = 8.0Hz), 7.29-7.33 (t, 2H, J = 8.0Hz), 7.21-7.26 ( m, 2H), 7.14 (s, 1H), 6.98-7.00 (d, 1H, J = 8.0Hz), 2.75 (s, 2H), 2.33 (s, 3H), 2.16 (s, 3H)

Compounds 2.2a and 2.2b

Each racemic 2.2a and 2.2b enantiomeric system was separated using preparative HPLC on CHIRALPAK ASH with CO ₂ /MeOH/DEA=70/30/0.1 as the eluent. The enantiomeric transcendence value was measured using CHIRALPAK ASH upper limb HPLC with CO ₂ /MeOH/DEA=60/40/0.1 (v / v / v) as the eluent, and the flow rate was measured at 2.0 mL / min. The retention time of the first enantiomer was 4.84 min, (DMSO- d ₆ ) δ _H 10.53 (s, 1H), 7.20-7.41 (m, 6H), 7.13 (d, J = 1.6Hz, 1H) , 6.99 (d, J = 8.0Hz, 1H), 2.71 (br s, 2H), 2.32 (s, 3H), 2.15 (s, 3H); the retention time of the other enantiomer is 6.91min, ( DMSO- d ₆ ) δ _H 10.53 (s, 1H), 7.20-7.41 (m, 6H), 7.13 (d, J = 2.0Hz, 1H), 6.99 (d, J = 8.0Hz, 1H), 2.71 (br s, 2H), 2.33 (s, 3H), 2.16 (s, 3H) .MS (ESI) m / e [M + 1] ⁺ 320.

Following similar procedures as described in compound 2.1 , the following compounds 2.3 to 2.37 were synthesized starting from the corresponding reagents.

Step 1: (S, Z) -N- (5- (3,5-dimethyliso Azole-4-yl) -2-oxoindol-3-yl) -2-methylpropane-2-sulfonamide

5- (3,5-dimethyliso (Zol-4-yl) dihydroindole-2,3-dione (2.42 g, 10 mmol) in THF (50 mL), (S) -2-methylpropane-2-sulfinamide ( 1.33 g, 11 mmol) and tetraethyl titanate (9.12 g, 40 mmol). The mixture was heated to reflux for 15 h, then cooled to room temperature, saturated sodium bicarbonate (50 mL) was added, filtered, washed with ethyl acetate (50 mL), and the filtrate was extracted with ethyl acetate (2 x 50 mL), dried over sodium sulfate, and filtered. , And concentrated under reduced pressure, and the mixture was purified by column chromatography to obtain a red solid (2.6 g, 74%). ¹ H NMR (400MHz, DMSO-d ₆ ): δ _H 10.80 (s, 1H), 7.57 (d, J = 8.0Hz, 2H), 7.18-7.32 (m, 5H), 6.93 (d, J = 8.0Hz , 1H), 4.66-4.68 (m, 1H), 4.15-4.29 (m, 2H), 2.85-2.89 (m, 5H), 2.37 (s, 3H), 2.18 (s, 3H), 1.91-1.98 (m , 1H), 1.63-1.68 (m, 1H). MS (ESI) m / e [M + 1] ⁺ 346.

Step 2: (S, Z) -Third-butyl 3- (Third-butylsulfonimide) -5- (3,5-dimethyliso Azole-4-yl) -2-oxoindolin-1-formate

(S, Z) -N- (5- (3,5-dimethylisopropyl Azole-4-yl) -2-oxoindolin-3-yl) -2-methylpropane-2-sulfonamide (0.50 g, 1.43 mmol), (Boc) ₂ O (0.38 g, 1.74 mmol) and DMAP (0.017 g, 0.14 mmol) in anhydrous THF (20 mL), and stirred from 0 ° C to room temperature for 2.5h. After cooling to 0 ° C, the reaction mixture was quenched by the addition of saturated sodium bicarbonate (10 mL). The mixture was extracted with dichloromethane (2 x 20 mL). The organic phases were combined, dried, filtered, and the filtrate was concentrated under reduced pressure. Further purification was used directly in the next step.

Step 3: (S) -Third-butyl-3-cyclohexyl-3- (1,1-dimethylethylsulfonamide) -5- (3,5-dimethyl-iso Azole-4-yl) -2-oxoindolin-1-formate

(S, Z) -Third-butyl3- (Third-butylsulfonimide) -5- (3,5-dimethyliso Azol-4-yl) -2-oxoindolin-1-formate (0.4 mmol) was added to 6 mL of anhydrous THF, and a cyclohexyl magnesium chloride solution (0.9 mmol) was added. The reaction mixture was stirred at 0 ° C for 1 h. Stir for 12h. The reaction was quenched with saturated aqueous ammonium chloride (10 mL), and the organic phase was extracted with dichloromethane (2 x 10 mL), dried over sodium sulfate, and evaporated under reduced pressure. The crude adduct was purified by flash chromatography to obtain the desired isomer as the main product. MS (ESI) m / e [M + 1-100] ⁺ 430.

Step 4: 3-Amino-3-cyclohexyl-5- (3,5-dimethyliso Azole-4-yl) indolin-2-one

(S) -Third-butyl-3-cyclohexyl-3- (1,1-dimethylethylsulfonamido) -5- (3,5-dimethyl-iso Azol-4-yl) -2-oxoindolin-1-formate (53 mg, 0.1 mmol) in a solution of dioxane (2 mL), and a saturated solution of dioxane in hydrochloric acid (1.0 mmol) and the reaction mixture was stirred for 15 min. The reaction was quenched with 6 mL of sodium bicarbonate aqueous solution, the organic phase was extracted with dichloromethane, and evaporated under reduced pressure to obtain pure (S) -3-amino-3-cyclohexyl-5- (3,5-dimethyliso Zol-4-yl) indolin-2-one (21 mg, 65%) was a solid. ¹ H NMR (400MHz, DMSO-d ₆ ): δ _H 10.32 (s, 1H), 7.17-7.19 (m, 2H), 6.88 (d, J = 8.8Hz, 1H), 2.39 (s, 3H), 2.21 (s, 3H), 1.90-1.93 (m, 1H), 1.53-1.63 (m, 5H), 0.95-1.11 (m, 4H), 0.68-0.72 (m, 1H) .MS (ESI) m / e [ M + 1] ⁺ 326.

3-amino-3-cyclohexyl-5- (3,5-dimethyliso Azole-4-yl) indolin-2-one

Compound 2.37b was synthesized using the same procedure as compound 2.37a . ¹ H NMR (400MHz, DMSO-d ₆ ): δ _H 10.35 (s, 1H), 7.18-7.20 (m, 2H), 6.89 (d, J = 8.4Hz, 1H), 2.39 (s, 3H), 2.21 (s, 3H), 1.90-1.93 (m, 1H), 1.53-1.68 (m, 5H), 0.95-1.11 (m, 4H), 0.68-0.72 (m, 1H) .MS (ESI) m / e ( M + 1] ⁺ 326.

Step 1: Third butyl-4- (5- (3,5-dimethylisopropyl) Azol-4-yl) -2-oxo-3-phenylindol-3-yl) piperazine -1-formate

676 mg (2.0 mmol) of 3-chloro-5- (3,5-dimethylisopropyl) at room temperature Azole-4-yl) -3-phenylindolin-2-one and 774 mg (6.0 mmol) of diisopropylethylamine were dissolved in 20 ml of tetrahydrofuran, and 774 mg (4.0 mmol) of tert-butyl Kippi After the 1-formate, the reaction solution was stirred for 0.5 hours. Then, the solution was diluted with water, and the aqueous phase was extracted with dichloromethane (2 × 30 mL). The organic phases were combined, washed with an aqueous solution of sodium bicarbonate and water, dried and concentrated under reduced pressure. CH ₂ Cl ₂ / MeOH = 20/1) was purified to give the product (940 mg, 96%) as a white solid. ¹ H NMR (400MHz, DMSO- d ₆ ): δ _H 10.75 (s, 1H), 7.51 (d, J = 8.0Hz, 1H), 7.25-7.38 (m, 6H), 6.98 (d, J = 8.0Hz , 1H), 3.27-3.29 (m, 4H), 2.47-2.38 (m, 4H), 2.36 (s, 3H), 2.18 (s, 3H), 1.35 (s, 9H) .MS (ESI) m / e [M + 1] ⁺ 489.

Step 2: 5- (3,5-Dimethyliso Azol-4-yl) -3-phenyl-3- (piperazine -1-yl) indol-2-one

At 0 ° C, the third butyl-4- (5- (3,5-dimethylisopropyl) Azol-4-yl) -2-oxo-3-phenylindol-3-yl) piperazine To a solution of 1-formate (0.9 g, 1.8 mmol) in dichloromethane (5 mL), TFA (5 mL) was added. After the dropwise addition, the reaction was stirred at room temperature for 2.0 h. The solvent was evaporated under reduced pressure to obtain a crude product. The crude product was dissolved in water (20 mL), neutralized with sodium bicarbonate to pH = 7-8, and the mixture was extracted with ethyl acetate (2 × 30 mL). The organic phases were combined to The sodium sulfate was dried, filtered, and concentrated to obtain the crude product, and the mixture was purified by chromatography (eluent: CH ₂ Cl ₂ / MeOH / NH ₃ H ₂ O = 20/1 / 0.05) to obtain the product (500 mg, 71.6%) As a white solid. ¹ H NMR (400MHz, DMSO-d6): δ _H 10.75 (s, 1H), 7.49 (d, J = 8.0Hz, 2H), 7.23-7.36 (m, 5H), 6.97 (d, J = 8.0Hz, 1H), 2.65-2.66 (m, 4H), 2.31-2.47 (m, 7H), 2.19 (s, 3H). MS (ESI) m / e [M + 1] ⁺ 389.

The following compounds 2.39 to 2.46 were synthesized starting from the corresponding reagents using similar procedures as described for compound 2.38 .

2- (4- (5- (3,5-dimethyliso Azol-4-yl) -2-oxo-3-phenylindol-3-yl) piperazine 1-yl) ethyl acetate

Remove 5- (3,5-dimethylisopropyl) at room temperature Azol-4-yl) -3-phenyl-3- (piperazine To a solution of 1-yl) indolin-2-one (194 mg, 0.5 mmol) in DMF (3.0 mL), ethyl bromoacetate (417 mg, 2.5 mmol) and potassium carbonate (208 mg, 1.5 mmol) were added, The mixture was stirred for 0.5 h, and then saturated ammonium chloride (10 mL) was added. The mixture was extracted with dichloromethane (2 × 10 mL). The organic phases were combined, dried over sodium sulfate, and concentrated under reduced pressure to obtain a crude product. The mixture was subjected to column chromatography. (using _{CH 2 Cl 2 / MeOH (50} /1, v / v) elution) to give a white solid (100mg, 42%). ¹ H NMR (400MHz, DMSO- d ₆ ): δ _H 7.52 (d, J = 8.0Hz, 2H), 7.29-7.37 (m, 5H), 7.17 (d, J = 8.0Hz, 1H), 4.65 (s , 2H), 4.16 (q, J = 6.8Hz, 2H), 2.50-2.52 (m, 4H), 2.36 (s, 3H), 2.18 (s, 3H), 1.17 (t, J = 6.8Hz, 3H) .MS (ESI) m / e [M + 1] ⁺ 475.

5- (3,5-dimethyliso Azol-4-yl) -3- (4- (2-hydroxyethyl) piper (1--1-yl) -3-phenylindol-2-one

At 0 ° C, 2- (4- (5- (3,5-dimethylisopropyl Azol-4-yl) -2-oxo-3-phenylindol-3-yl) piperazine To a solution of 1-yl) ethyl acetate (100 mg, 0.21 mmol) in THF (5 mL) was added lithium tetrahydroaluminum (38 mg, 1.05 mmol), and the reaction mixture was stirred at room temperature for 2.0 h. After cooling to 0 ° C, the reaction was quenched by adding a saturated aqueous ammonium chloride solution (50 mL), the mixture was diluted with water and extracted with ethyl acetate (50 mL), the organic phase was washed with brine, dried over anhydrous magnesium sulfate, filtered and evaporated under reduced pressure. A crude product was produced, which was purified by flash column chromatography to give the title compound as a white solid (30 mg, 33%). ¹ H NMR (400MHz, DMSO- d ₆ ): δH 10.69 (s, 1H), 7.22-7.51 (m, 7H), 6.96 (d, J = 8.0Hz, 1H), 4.31 (s, 1H), 3.40- 3.45 (m, 2H), 2.13-2.40 (m, 13H), 2.02 (s, 3H). MS (ESI) m / e [M + 1] ⁺ 433.

5- (3,5-dimethyliso Azol-4-yl) -3- (4- (3-hydroxypropyl) piper (1--1-yl) -3-phenylindol-2-one

Remove 5- (3,5-dimethylisopropyl) at room temperature Azol-4-yl) -3-phenyl-3- (piperazine -1-yl) indol-2-one (77 mg, 0.2 mmol), 3-bromopropyl-1-ol (278 mg, 2.0 mmol) and potassium carbonate (55 mg, 0.4 mmol) in DMF (3 mL) The mixture was stirred for 0.5 h, then diluted with ethyl acetate (20 mL) and extracted with water (2 x 20 mL) and brine (50 mL). The aqueous layer was washed again with ethyl acetate (20 mL). The organic layers were combined and dried (magnesium sulfate ), Filtered and concentrated, the residue was purified by Pre-TLC (using CH ₂ Cl ₂ / MeOH (10: 1) as the eluent) to obtain the product (20 mg, 22%) as a white solid. ¹ H NMR (400MHz, DMSO- d ₆ ): δH 10.69 (s, 1H), 7.23-7.50 (m, 7H), 6.96 (d, J = 8.0Hz, 1H), 3.36-3.39 (m, 2H), 2.29-2.32 (m, 13 H), 2.18 (s, 3H), 1.50-1.52 (m, 2H) .MS (ESI) m / e [M + 1] ⁺ 447.

Add 3-chloro-5- (3,5-dimethyliso Azole-4-yl) -3-phenylindolin-2-one 676 mg (2.0 mmol) and (2S, 4R) -methyl-4-hydroxypyrrolidin-2-carboxylic acid methyl ester 544 mg (3.0 mmol ) Was dissolved in dichloromethane (10 mL), and the reaction solution was stirred at room temperature for 0.5 h. Then, the solution was diluted with water, and the aqueous phase was extracted with dichloromethane (2 × 10 mL). The combined organic phases were washed with an aqueous sodium hydrogen carbonate solution and water, dried and concentrated under reduced pressure. The obtained residue was chromatographed (eluent: CH ₂ Cl ₂ / MeOH = 50/1) was purified to obtain a white solid with fast reaction isomer product (150 mg, 16%) and slow reaction isomer product (140 mg, 15%) in normal phase column chromatography. . Fast reaction isomers ( 2.50a ): ¹ H NMR (400 MHz, DMSO- d ₆ ): δ _H 10.85 (s, 1H), 7.50 (d, J = 8.0 Hz, 2H), 7.20-7.36 (m, 4H), 7.16 (s, 1H), 6.94 (d, J = 8.0Hz, 1H), 4.80 (d, J = 5.2Hz, 1H), 4.21-4.25 (m, 1H), 3.50-3.54 (m, 1H ), 3.14 (s, 3H), 3.05-3.09 (m, 1H), 2.87-2.91 (m, 1H), 2.36 (s, 3H), 2.16 (s, 3H), 1.85-1.93 (m, 2H), MS (ESI) m / e [M + 1] ⁺ 448; Slow reaction isomer ( 2.50b ): ¹ H NMR (400MHz, DMSO- d ₆ ): δ _H 10.82 (s, 1H), 7.50 (d , J = 8.0Hz, 2H), 7.17-7.30 (m, 5H), 6.93 (d, J = 8.0Hz, 1H), 4.91 (d, J = 4.8Hz, 1H), 4.26-4.31 (m, 1H) , 4.15-4.18 (m, 1H), 3.34 (s, 3H), 3.10-3.34 (m, 1H), 2.31-2.35 (m, 4H), 2.16 (s, 3H), 1.86-1.90 (m, 2H) , MS (ESI) m / e [M + 1] ⁺ 448.

Compounds 2.51a and 2.51b

Using the same procedure described for compound 2.48 , compounds 2.51a and 2.51b were synthesized from compounds 2.50a and 2.50b . Fast reaction isomer ( 2.51a ): ¹ H NMR (400MHz, DMSO- d ₆ ): δ H 10.68 (s, 1H), 7.23-7.58 (m, 6H), 6.95 (d, J = 7.6Hz, 1H), 4.70 (d, J = 4.8Hz, 1H), 4.13-4.16 (m, 2H), 2.89-3.00 (m, 4H), 2.38 (s, 3H), 2.29-2.33 (m, 1H), 2.21 (s, 3H), 1.87-1.92 (m, 1H), 1.55-1.59 (m, 1H) .MS (ESI) m / e [M + 1] ⁺ 420; Slow reaction isomer ( 2.51b ): ¹ H NMR (400MHz, DMSO-d ₆ ): δ _H 10.80 (s, 1H), 7.57 (d, J = 8.0Hz, 2H), 7.18-7.32 (m, 5H), 6.93 (d, J = 8.0Hz , 1H), 4.66-4.68 (m, 1H), 4.15-4.29 (m, 2H), 2.85-2.89 (m, 5H), 2.37 (s, 3H), 2.18 (s, 3H), 1.91-1.98 (m , 1H), 1.63-1.68 (m, 1H). MS (ESI) m / e [M + 1] ⁺ 420.

The following compounds 2.52 to 2.58 were synthesized starting from the corresponding reagents using similar procedures as described for compounds 2.50a and 2.51a .

Step 1: 5-Bromo-3- (4-fluorophenyl) -3-hydroxyindoline-2-one

In a solution of 5-bromodihydroindole-2,3-dione (120g, 530mmol, 1.0eq) in THF (1L), (4-fluorophenyl) magnesium bromide was added dropwise at 0 ° C to 15 ° C. The mixture was stirred at THF (0.8M, 1.6 L, 1280 mmol, 2.4 eq) at room temperature overnight. The mixture was then cooled to 0 ° C, quenched with water (20 mL), and concentrated to give the crude product, which was dissolved in ethyl acetate (2L), washed with 1N aqueous hydrochloric acid solution (2L), brine (1L x 2), and sulfuric acid The crude product was obtained after drying with sodium and concentrating the organic phase. The crude product was then stirred in PE, and the resulting yellow solid was filtered. After drying in air, 136 g (yield 80%) of the product was obtained without further purification. Used for the next step. ¹ H NMR (400MHz, DMSO- d ₆ ): δ _H 10.58 (s, 1H), 7.44-7.46 (dd, 1H, J ₁ = 8.4Hz, 2.0Hz), 7.28-7.32 (m, 2H), 7.22- 7.25 (m, 1H), 7.14-7.18 (t, 2H, J = 8.8Hz), 6.87-6.89 (d, 1H, J = 8.4Hz), 6.84 (s, 1H) .MS (ESI) (M + 1 -18] ⁺ 304,306.

Step 2: 5- (3,5-Dimethyliso Azole-4-yl) -3- (4-fluorophenyl) -3-hydroxyindol-2-one

3,5-dimethyliso Azole-4-ylboronic acid (120 g, 851 mmol, 2.0 eq), dichloro [1,1'-bis (di-third-butylphosphino) ferrocene] palladium (II) (10 g, 13.7 mmol, 0.03 eq ), A mixture of 5-bromo-3- (4-fluorophenyl) -3-hydroxyindolin-2-one (136 g, 422 mmol, 1.0 eq) and sodium carbonate (100 g, 943 mmol, 2.2 eq) was dissolved Dioxane / water (800 mL / 200 mL) was heated under reflux in a nitrogen environment for 5 hours. After cooling to room temperature, 1 liter of EA was added, and the mixture was filtered through a pad of celite, the filtrate was washed with brine (1 L × 2), dried over sodium sulfate, concentrated, and purified by silica gel to obtain 70 g of a yellow solid (yield 49 %). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ _H 10.56 (s, 1H), 7.31-7.33 (m, 2H), 6.93 (d, J = 8.0 Hz, 1H), 4.27-4.35 (m, 4H ), 2.35 (s, 3H), 2.18 (s, 3H) .MS (ESI) m / e [M + 1] ⁺ 339.

Step 3: 3-Amino-5- (3,5-dimethyliso Azole-4-yl) -3- (4-fluorophenyl) indol-2-one

Under nitrogen, remove 5- (3,5-dimethyliso Azole-4-yl) -3- (4-fluorophenyl) -3-hydroxyindolin-2-one (52g, 154mmol, 1.0eq) in a solution of THF (500mL) at -15 ° C ~ Pyridine (40 mL, 496 mmol, 3.2 eq) and thallium chloride (16 mL, 219 mmol, 1.4 eq) were slowly added at -20 ° C. The mixture was stirred at -15 ° C to -20 ° C for 20 minutes, quenched with brine, and then brine (300 mL). × 2) Washing, NH ₃ / H ₂ O (100 ml) was added to the tetrahydrofuran layer, and the mixture was stirred at room temperature for 2 hours, washed with brine, dried, concentrated, and purified with silica gel to obtain a crude product, which was subjected to isopropanol / Toluene recrystallized to give 14 g (27% yield) of the product as a white solid. ¹ H NMR (DMSO-d ₆ ) δ _H 10.57 (s, 1H), 7.40-7.44 (m, 2H), 7.22-7.24 (dd, 1H, J = 8.0Hz, 1.6Hz), 7.11-7.16 (m, 3H), 6.98-7.00 (d, 1H, J = 8.0 Hz), 2.75 (s, 2H), 2.33 (s, 3H), 2.16 (s, 3H).

Each racemic 2.59a and 2.59b enantiomeric system was separated using CHIRALPAK OJ-H preparative HPLC with carbon dioxide / (ethanol 80 acetonitrile 20) = 72/28 as the eluent. The enantiomeric excess was determined by HPLC of CHIRALPAK OJ-H. Carbon dioxide / (methanol 70 acetonitrile 30) = 70/30 was used as the eluent, and the flow rate was 2.0 mL / min. The retention time of the first enantiomeric elution was 3.55 min, ¹ H NMR (400MHZ, DMSO- D ₆ ): δ _H 10.57 (s, 1H), 7.11-7.44 (m, 6H), 6.98 (d, J = 7.6HZ, 1H), 2.76 (s, 2H), 2.34 (s, 3H), 2.17 (s, 3H) .MS (ESI) M / E [M + 1] ⁺ 338; the other enantiomer was eluted Retention time is 7.07min, ¹ H NMR (400MHZ, DMSO- D ₆ ): δ _H 10.55 (s, 1H), 7.11-7.44 (m, 6H), 6.98 (d, J = 8.0HZ, 1H), 2.76 (s, 2H), 2.33 (s, 3H), 2.16 (s, 3H) .MS (ESI) M / E [M + 1] ⁺ 338.

Example 3: Synthesis of compounds 3.1 to 3.10

5- (3,5-dimethyliso Azole-4-yl) -3-phenylindol-2-one

5- (3,5-dimethyliso Triazole-4-yl) -3- (2-hydroxyethylamino) -3-phenylindolin-2-one (32 g, 0.1 mmol) in dichloromethane (500 mL) was added with trifluoroacetic acid (20 g) and triethylsilane (20 g). The brown solution was stirred at ambient temperature for 3h, and concentrated to dryness under reduced pressure. The residue was diluted with dichloromethane (500mL), washed with saturated ammonium chloride solution (200mL), brine (3 × 400mL), dried over anhydrous sodium sulfate, and After filtration, the filtrate was concentrated to dryness under reduced pressure, and the residue was crystallized from diethyl ether to obtain the target compound. ¹ H NMR (400MHz, DMSO- d ₆ ): δ _H 10.63 (s, 1H), 7.18-7.36 (m, 6H), 6.99-7.04 (m, 2H), 4.82 (s, 1H), 2.32 (s, 3H), 2.15 (s, 3H). MS (ESI) m / e [M + 1] ⁺ 305.

5- (3,5-dimethyliso Azole-4-yl) -3- (3-hydroxypropyl) -3-phenylindol-2-one

In a covered pressure tube, 5- (3,5-dimethylisopropyl Azole-4-yl) -3-phenylindolin-2-one (1.5 g, 5 mmol), 3-bromoprop-1-ol (1.3 g, 10.0 mmol), potassium iodide (0.16 g, 1 mmol) and A mixture of potassium carbonate (1.3 g, 10.0 mmol) in THF (50 mL) was heated at 60 ° C for 5 hours. The mixture was then cooled to room temperature, diluted with ethyl acetate (20 mL), and extracted with water (2 x 20 mL) and brine (20 mL). The aqueous layer was washed again with ethyl acetate (20 mL), the organic layers were combined, dried (MgSO ₄ ), filtered and concentrated. The residue was subjected to flash chromatography (dichloromethane, then 5% ethyl acetate in dichloromethane as the solvent). ) Purification gave the product (0.74 g, 41%). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ _H 10.65 (s, 1H), 7.23-7.36 (m, 7H), 7.00 (d, J = 8.0 Hz, 1H), 4.39 (m, 1H), 3.31 -3.33 (m, 2H), 2.37 (s, 3H), 2.18-2.28 (m, 2H), 2.16 (s, 3H), 1.21-1.27 (m, 1H), 1.02-1.06 (m, 1H) .MS (ESI) m / e [M + 1] ⁺ 363.

2- (5- (3,5-dimethyliso Azol-4-yl) -2-oxo-3-phenylindol-3-yl) ethyl acetate

In a covered pressure tube, 5- (3,5-dimethylisopropyl Azol-4-yl) -3-phenylindol-2-one (260 mg, 0.85 mmol), ethyl bromoacetate (172 mg, 1.0 mmol), potassium iodide (171 mg, 1.0 mmol) and potassium carbonate (260 mg, A mixture of 1.88 mmol) in acetone (10 mL) was heated at 60 ° C for 15 hours. The mixture was then cooled to room temperature and diluted with ethyl acetate (30 mL) and extracted with water (2 x 30 mL) and brine (20 mL). The aqueous layer was washed again with ethyl acetate (30 mL), the organic phases were combined, dried (MgSO ₄ ), filtered and concentrated. The residue was subjected to flash chromatography (dichloromethane, then 5% ethyl acetate in dichloromethane as a solvent). ) Purified to give the product (200 mg, 59.9%). ¹ H NMR (400 MHz, DMSO- d ₆ ): δ _H 10.61 (s, 1H), 7.21-7.39 (m, 7H), 6.97 (d, J = 8.0 Hz, 1H), 3.82-3.85 (m, 2H) , 3.41 (s, 3H), 2.36 (s, 3H), 2.19 (s, 3H), 0.90 (t, J = 6.8Hz, 1H) .MS (ESI) m / e [M + 1] ⁺ 391.

2- (5- (3,5-dimethyliso Azol-4-yl) -2-oxo-3-phenylindol-3-yl) acetic acid

Add 2- (5- (3,5-dimethyliso Ethyl-4-yl) -2-lanthoxy-3-phenylindol-3-yl) ethyl acetate (35 mg, 0.089 mmol) was dissolved in THF / MeOH (2 mL / 2 mL), and LiOH was added. H ₂ O (40 mg) in H ₂ O (2 mL) and the reaction was stirred at room temperature until the starting material indicated by TLC was completely consumed. Water was added to the reaction, acidified to pH <1 with 3% hydrochloric acid, and extracted with ethyl acetate (3 × 5 mL). The combined extracts were washed with brine, dried over MgSO ₄ and concentrated under reduced pressure. The title compound was purified by Pre-TLC Purification gave the title compound (20 mg, 62%) as a white solid. ¹ H NMR (400MHz, DMSO- d ₆ ) δ _H 12.17 (s, 1H), 10.56 (s, 1H), 7.41 (s, 1H), 7.35-7.29 (m, 4H), 7.22-7.24 (m, 2H ), 6.96 (d, J = 8.0Hz, 1H), 3.29-3.30 (m, 2H), 2.38 (s, 3H), 2.21 (s, 3H) .MS (ESI) m / e [M + 1] ⁺ 363.

5- (3,5-dimethyliso Azole-4-yl) -3- (2-hydroxyethyl) -3-phenylindol-2-one

Methyl 2- (5- (3,5-dimethyliso Azol-4-yl) -2-oxo-3-phenylindol-3-yl) acetate (50 mg, 0.13 mmol) in ethanol (20 mL) was added to sodium borohydride (49 mg, 1.3 mmol) and the reaction mixture was stirred to reflux for 3.0 h. After cooling to 0 ° C, the reaction was quenched by the addition of a saturated aqueous ammonium chloride solution (2 mL), the mixture was diluted with water (10 mL), and extracted with ethyl acetate (15 mL). The organic layer was washed with brine, dried over anhydrous MgSO ₄ , and filtered. The crude product was evaporated under reduced pressure and purified by flash column chromatography to give the title compound as a white solid (10 mg, 22%). ¹ H NMR (400MHz, DMSO- d ₆ ): δ _H 10.59 (s, 1H), 7.22-7.35 (m, 7H), 6.98 (d, J = 8.0 Hz, 1H), 4.48-4.51 (m, 1H) , 3.15-3.17 (m, 2H), 2.44-2.48 (m, 2H), 2.32 (s, 3H), 2.20 (s, 3H). MS (ESI) m / e [M + 1] ⁺ 349.

Compound 3.6

5- (3,5-dimethyliso Azol-4-yl) -3- (3- (Phenylpropyl) -3-phenylindol-2-one

Compound 5- (3,5-dimethyliso Azol-4-yl) -3- (3-hydroxypropyl) -3-phenylindol-2-one (0.72 g, 2.0 mmol), DIPEA (1.05 mL, 6.0 mmol), MsCl (250 mg, A mixture of 2.2 mmol) in CH ₂ Cl ₂ (20 mL) was stirred at 0 ° C. and then gradually raised to room temperature overnight. The content was diluted with dichloromethane, washed with water (2 x 10 mL), and concentrated. The crude product was purified by flash chromatography to give compound 3- (5- (3,5-dimethyliso Azol-4-yl) -2-oxo-3-phenylindol-3-yl) propyl methanesulfonate (575 mg, 65.3%). ¹ H NMR (400MHz, DMSO- d ₆ ): δ _H 10.73 (s, 1H), 7.24-7.38 (m, 7H), 7.02 (d, J = 8.0Hz, 1H), 4.13-4.16 (m, 2H) , 3.12 (s, 3H), 2.37 (s, 3H), 2.30-2.34 (m, 2H), 2.20 (s, 3H), 1.30-1.48 (m, 2H). 3- (5- (3,5-dimethylisopropyl) at room temperature Azole-4-yl) -2-oxo-3-phenylindol-3-yl) propyl propyl mesylate (440 mg, 1.0 mmol) in a stirred solution of anhydrous tetrahydrofuran (10 mL), in that order Add triethylamine (280 μl, 2.0 mmol) and Phorphyrin (174mg, 2.0mmol), the reaction mixture was heated at 80 ° C for 15h, then water (10mL) was added, and the aqueous layer was extracted with dichloromethane (3 × 10mL). The combined organic phases were dried over sodium sulfate and filtered. After concentration under pressure, the residue was purified by silica gel chromatography (eluent: CH ₂ Cl ₂ / MeOH: 95/5) to obtain the compound (385 mg, 89.3%) as a white solid. ¹ H NMR (400MHz, CDCl ₃ ): δ _H 7.88 (s, 1H), 7.28-7.39 (m, 5H), 7.14-7.16 (m, 1H), 7.08 (s, 1H), 7.02 (d, J = 8.0Hz, 1H), 3.76 (m, 4H), 2.45-2.55 (m, 5H), 2.37-2.39 (m, 4H), 2.22-2.29 (m, 5H), 1.54-1.55 (m, 2H) .MS (ESI) m / e [M + 1] ⁺ 432.0.

The following compounds 3.7 to 3.10 were synthesized starting from the corresponding reagents using similar procedures as described for compound 3.6 .

Example 4: Synthesis of compounds 4.1 to 4.11

5- (3,5-dimethyliso Azol-4-yl) -3-hydroxy-3- (thien-3-yl) indol-2-one and 5- (3,5-dimethyliso Azole-4-yl) -3-hydroxy-3- (2-oxopropyl) indol-2-one

In a direct-fired dry Schlenk bottle filled with nitrogen, 145 mg (0.56 mmol, 3 mol%) of Rh (acac) (C ₂ H ₄ ) ₂ and 405 mg (1.31 mol, 7 mol%) of triphenylphosphine were dissolved in 200 mL. After stirring at room temperature for 5 min, 4.54 g (18.75 mmol) of the substrate 5- (3,5-dimethylisopropyl) was added. Azole-4-yl) indol-2,3-dione and 4.8 g (37.5 mmol) of thiophene-3-boronic acid. The resulting mixture was stirred to reflux temperature. After 40 h, the reaction mixture was cooled to room temperature and reduced in temperature. The solvent was evaporated under pressure. Purification by column chromatography using the elution conditions reported by TLC gave two products as white solids whose structures were determined by NMR and LC / MS. Among them, the fast-reacting compound is 5- (3,5-dimethyliso Azol-4-yl) -3-hydroxy-3- (2-oxopropyl) -dihydroindole-2-one: ¹ H NMR (400 MHz, DMSO- d ₆ ): δ _H 10.33 (s, 1H ), 7.26 (d, J = 1.6 Hz, 1H), 7.18 (dd, J = 1.6, 8.0 Hz, 1H), 6.80 (d, J = 8.0 Hz, 1H), 6.12 (s, 1H), 3.35 (d , J = 16.4Hz, 1H), 3.01 (d, J = 16.4Hz, 1H), 2.27 (s, 3H), 2.19 (s, 3H), 2.02 (s, 3H) .MS (ESI) m / e ( M + 1] ⁺ 301; the slower reacting compound is 5- (3,5-dimethyliso Azole-4-yl) -3-hydroxy-3- (thiophen-3-yl) indol-2-one: ¹ H NMR (400 MHz, DMSO- d ₆ ): δ _H 10.47 (s, 1H), 7.47 (dd, J = 4.8, 3.2Hz, 1H), 7.22-7.26 (m, 3H), 7.13 (d, J = 4.8Hz, 1H), 6.96 (d, J = 8.0Hz, 1H), 6.65 (s , 1H), 2.36 (s, 3H), 2.19 (s, 3H). MS (ESI) m / e [M + 1] ⁺ 327.

Examples 4.1a and 4.1b

Compound 4.1 was separated by chiral prep-HPLC to obtain two enantiomeric stereoisomers (compound 4.1a , a fast-reacting isomer; and compound 4.1b , a slow-reaction isomer). Chiral separation conditions are as follows:

Example 4.3

Step 1: 3-Chloro-5- (3,5-dimethyliso Azole-4-yl) -3- (thien-3-yl) indol-2-one

5- (3,5-dimethylisopropyl) at 0 ℃ Azol-4-yl) -3-hydroxy-3- (thien-3-yl) indolin-2-one (500 mg, 1.53 mmol) in a solution of dichloromethane (75.0 mL), and pyridine (1.21 g, 15.3 mmol), then dichloromethane (728 mg, 6.12 mmol) was added, and the reaction mixture was stirred at 0 ° C. for 2 h. The mixture was evaporated under reduced pressure to obtain a crude product, which was used in the next step without further purification.

Step 2: 5- (3,5-Dimethyliso Azole-4-yl) -3- (2-hydroxyethylamino) -3- (thien-3-yl) indol-2-one

3-chloro-5- (3,5-dimethyliso 110 mg (0.33 mmol) of diazol-4-yl) -3- (thien-3-yl) dihydroindol-2-one and 220 mg (1.65 mmol) of DIPEA were dissolved in 25 ml of dichloromethane, and 2-aminoethanol was added After 55 mg (0.65 mmol), the reaction solution was stirred at room temperature for 16 h. Then, the solution was diluted with water, and the aqueous phase was extracted with dichloromethane (2 × 20 mL). The organic phases were combined, washed with an aqueous sodium hydrogen carbonate solution and water, dried and concentrated under reduced pressure. The obtained residue was subjected to chromatography (eluent : CH ₂ Cl ₂ / MeOH = 20/1) was purified to give the product (85 mg, 69.8%) as a white solid. ¹ H NMR (400MHz, DMSO- d ₆ ): δH 7.48-7.50m, 1H), 7.20-7.31 (m, 3H), 6.97 (d, J = 8.0Hz, 1H), 4.47 (t, J = 5.6Hz , 1H), 3.20-3.38 (m, 4H), 3.17 (d, J = 5.3Hz, 2H), 2.37 (s, 3H), 2.20 (s, 3H) .MS (ESI) m / e (M + 1 ] ⁺ 370.

The following compounds 4.4 to 4.10b were synthesized starting from the corresponding reagents using similar procedures as described for compound 4.3 .

Example 4.11

5- (3,5-dimethyliso Azole-4-yl) -3- (thien-3-yl) indol-2-one

5- (3,5-dimethyliso Triazole-4-yl) -3-hydroxy-3- (thien-3-yl) indol-2-one (50 mg, 0.153 mmol) in a solution of trifluoroacetic acid (10 mL), and triethylsilane was added (5 mL). The brown solution was stirred at ambient temperature for 3 h and concentrated to dryness under reduced pressure. The residue was diluted with dichloromethane (20 mL), washed with saturated ammonium chloride solution (10 mL) and brine (3 x 20 mL), dried over anhydrous sodium sulfate and After filtration, the filtrate was concentrated to dryness under reduced pressure, and the residue was purified by column chromatography to obtain the target compound (37 mg, 78.0%). 1H NMR (400MHz, DMSO-d6): δ _H 10.62 (s, 1H), 7.53 (m, 1H), 7.31-7.32 (m, 1H), 7.22 (d, J = 8.0Hz, 1H), 7.16 (s , 1H), 7.02-7.03 (m, 1H), 6.97 (d, J = 8.0Hz, 1H), 4.90 (s, 1H), 2.35 (s, 3H), 2.18 (s, 3H) .MS (ESI) m / e [M + 1] ⁺ 311.

Example 5.1: Synthesis of compound 5.1

Step 1: N- (4-Bromo-2-methoxyphenyl) -2- (hydroxyimino) acetamidamine

To a suspension of 2,2,2-trichloro-1-ethoxyethanol (6.96 g, 36 mmol) and sodium sulfate (38.3 g, 270 mmol) in water (45 mL) and 2N hydrochloric acid (30 mL) was added hydroxylamine hydrochloride (6.67 g, 96 mmol) in water (15 mL), and the mixture was stirred at 60 ° C. for 20 min. 4-Bromo-2-methoxyaniline (6.06 g, 30 mmol) was added to 2N hydrochloric acid (30 mL), and the mixture was heated at 90 ° C. for 2 h. The mixture was cooled to room temperature, filtered, washed with water, and dried in air to give the target compound (5.55 g, crude product, yield 68%) as a solid, which was used in the next step without further purification. MS (ESI) m / e [M + 1] ⁺ 273,275.

Step 2: 5-Bromo-7-methoxyindoline-2,3-dione

At 65 ° C, concentrated sulfuric acid (30ml) was added in several portions and N- (4-bromo-2-methoxyphenyl) -2- (hydroxyimino) acetamidamine (5.55g, 20mmol) was added. Heated at ℃ for 1.5h. The mixture was cooled to room temperature, poured into ice and stirred for 10 min, filtered, washed with water, and dried in air to give 5-bromo-7-methoxy-2,3-dione (3.6 g, crude product, yield 70%) of solid, this material was used in the next step without further purification. ¹ H NMR (400 MHz, DMSO- d ₆ ): δ _H 7.93 (s, 1H), 7.38 (s, 1H), 3.94 (s, 3H). MS (ESI) m / e [M + 1] ⁺ 256,258.

Step 3: 5'-Bromo-7'-methoxyspiro [[1,3] dioxolane-2,3'-dihydroindole] -2'-one

A mixture of 5-bromo-7-methoxyindol-2,3-dione (3.6 g, 14 mmol), p-toluenesulfonic acid (1.14 g, 6 mmol) and toluene (100 mL) was stirred, and Dean-Stark was used. The device was refluxed for 0.2 hours. Ethylene glycol (9.3 g, 150 mmol, 10.7 eq.) Was added and refluxed for another 2 hours using a Dean-Stark apparatus. Mixture was cooled and concentrated to 20mL, added ethyl acetate (100 mL), water (20mL) and brine (50mL × 2), dried over sodium sulfate, filtered, and concentrated under reduced pressure, by column (CH ₂ Cl ₂ / EtOAc = 8: 1) Purification gave the desired product as a light brown solid (0.21 g, crude product, 5%). ¹ H NMR (400MHz, DMSO- d ₆ ): δ _H 10.66 (s, 1H), 7.48 (s, 1H), 7.26-7.27 (m, 1H), 7.13-7.14 (m, 1H), 4.25-4.31 ( m, 4H), 3.86 (s, 3H). MS (ESI) m / e [M + 1] ⁺ 300/302.

Step 4: 5 '-(3,5-dimethylisopropyl Azole-4-yl) -7'-methoxyspiro [[1,3] dioxolane-2,3'-dihydroindole] -2'-one

3,5-dimethyliso Azole-4-boronic acid (298 mg, 2.1 mmol), dichloro [1,1'-bis (di-third-butylphosphino) ferrocene] palladium (II) (29 mg, 0.14 mmol), 5'-bromo- 7'-methoxyspiro [[1,3] dioxolane-2,3'-dihydroindole] -2'-one (210 mg, 0.7 mmol) and sodium carbonate (233 mg, 2.1 mmol) A mixture of oxane (9 mL) and water (2 mL) was heated at 102 ° C for 16 h. To the mixture were added ethyl acetate (50 mL) and water (20 mL). The organic layer was separated and washed with water (20 mL) and brine (10 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. Purification by column chromatography (using 33% EtOAc in dichloromethane as the eluent) gave the desired product as a light brown solid (57 mg, crude product, 26%). ¹ H NMR (400MHz, DMSO- d ₆ ): δ _H 10.62 (s, 1H), 7.02-7.03 (m, 1H), 6.92-6.93 (m, 1H), 4.26-4.34 (m, 4H), 3.86 ( s, 3H), 2.38 (s, 3H), 2.10 (s, 3H) .MS (ESI) m / e [M + 1] ⁺ 317.

Step 5: 5- (3,5-Dimethyliso Azole-4-yl) -7-methoxyindoline-2,3-dione

Compound 5 '-(3,5-dimethyliso Azol-4-yl) -7'-methoxyspiro [[1,3] dioxolane-2,3'-dihydroindole] -2'-one (57mg, 0.18mmol), acetic acid (1mL solution) and hydrochloric acid (4mL) was heated to the 90 ℃, after 1.5h, the mixture was cooled, ethyl acetate (20 mL), washed with brine (15mL × 3), dried over Na ₂ SO _4, and concentrated under reduced pressure to afford the desired Compound 5- (3,5-dimethyliso Azol-4-yl) -7-methoxyindoline-2,3-dione as a dark red solid (50 mg, crude product, 102%). This material was used in the next step without further purification. MS (ESI) m / e [M + 1] ⁺ 273.

Step 6: 5- (3,5-Dimethyliso Azole-4-yl) -3-hydroxy-7-methoxy-3-phenylindol-2-one

5- (3,5-dimethyliso A suspension of azole-4-yl) -7-methoxyindoline-2,3-dione (45 mg, 0.165 mmol) and anhydrous tetrahydrofuran (1.5 mL) was added dropwise to phenylmagnesium bromide at A solution of 2-methyltetrahydrofuran (2.9M / L, 0.3mL, 0.87mmol), and the reaction mixture was stirred at room temperature for 3h. The reaction mixture was diluted with ethyl acetate (30 mL), washed with saturated aqueous ammonium chloride solution (10 mL), brine (3 × 10 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure, and pre-TLC (CH ₂ Cl ₂ : EtOAc = 3: 2) purification gave the desired product as a light brown solid (28 mg, 48%). ¹ H NMR (400MHz, DMSO- d ₆ ): δ _H 10.56 (s, 1H), 7.31-7.32 (m, 5H), 6.95-6.96 (m, 1H), 6.69-6.71 (m, 2H), 3.89 ( s, 3H), 2.35 (s, 3H), 2.18 (s, 3H). MS (ESI) m / e [M + 1] ⁺ 351.

Example 6: Synthesis of compound 6.0

2- (dimethylamino) - N - (5- (3,5- dimethyl iso Azol-4-yl) -2-oxo-3-phenylindol-3-yl) acetamidamine

A mixture of 2- (dimethylamino) acetic acid hydrochloride (87 mg, 0.62 mmol), HATU (118 mg, 0.31 mmol) and triethylamine (94 mg, 0.93 mmol) in dichloromethane (10 mL) at room temperature Stir for 0.5 hours, then add 3-amino-5- (3,5-dimethyliso Zol-4-yl) -3-phenylindol-2-one (100 mg, 0.31 mmol), and the mixture was stirred for another 12 h. Water (10 mL) was added, and the mixture was extracted with dichloromethane (2 × 20 mL). The organic phases were combined, washed with brine (20 mL), dried over Na ₂ SO ₄ and concentrated under reduced pressure. The mixture was purified by column chromatography to obtain the product. (50 mg, 40%) was a solid. ¹ H NMR (400MHz, DMSO- d ₆ ): δ _H 10.70 (s, 1H), 9.74 (s, 1H), 7.26-7.40 (m, 7H), 6.99 (d, J = 8.0Hz, 1H), 4.07 (d, J = 16.0Hz, 1H), 3.93 (d, J = 16.0Hz, 1H), 2.78 (s, 3H), 2.71 (s, 3H), 2.38 (s, 3H), 2.20 (s, 3H) .MS (ESI) m / e [M + 1] ⁺ 405.

BRD2, BRD3, BRD4 and BRDT biochemical ICs ₅₀ Determination

The effects of the compounds in this case on BRD2, BRD3, BRD4 and BRDT were all determined by the time-resolved fluorescence resonance energy transfer (TR-FRET) method. Recombinant human BRD2 (1-473), BRD3 (1-435), BRD4 (1-477), and BRDT (1-397) with His tag at the N-terminus were expressed and purified by E. coli . Based assay containing 25mM HEPES pH 7.5,100mM NaCl, buffer, 0.1% BSA, 0.05% CHAPS and detection reagents bromodomain binding protein, 0-10 μ M with four compounds acetylated histone peptide (SGRG _AC- KGG _AC- KGLG _AC- KGGA _AC- KRHGSGSK-Biotin). When the protein, compound, and peptide have reached the binding equilibrium, a detection reagent containing streptavidin-labeled Tb cave compound and XL665-labeled anti-6xHis antibody is added. After further incubation for 1 hour, the TR-FRET signal was recorded with a BMG PHERAstar FS instrument. The four-parameter nonlinear regression equation of Graphpad Prism software is used: Y = Bottom + (Top-Bottom) / (1 + 10 ^ ((LogIC ₅₀ -X) * HillSlope)) to fit% INH, and then obtain the IC ₅₀ . Y is the percentage inhibition rate of the compound at the concentration X, X is the logarithm of the compound concentration, Bottom is the lowest value of the curve, Top is the highest value of the curve, and HillSlope is the hill slope factor.

Compounds 1.1 to 6.0 inhibited BRD2 / BRD3 / BRD4 / BRDT with IC ₅₀ values ranging from 0.1 nM to 10 μM.

Claims (10)

A compound of formula I, Or a pharmaceutically acceptable salt thereof, wherein: R ¹ is hydrogen, a halide, -OR ⁴ or -NR ⁵ R ⁶ , or a hydrocarbon group, and the hydrocarbon group is selected from C1-C8 alkyl, C2-C8 alkenyl, C2-C8 Alkynyl, C3-C8 cycloalkyl, C5-C8 cycloalkenyl, and C6-C14 aryl, in which each alkyl, alkenyl, and alkynyl group is selectively cyclized, and each hydrocarbon group is selectively substituted And optionally contains 1 to 3 heteroatoms, wherein R ⁴ , R ⁵ and R ^{6 are} each independently H or a hydrocarbon group, and the hydrocarbon group is selected from C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, And C6-C14 aryl groups, in which each alkyl, alkenyl and alkynyl group is selectively cyclized, and each hydrocarbon group is selectively substituted, and optionally contains 1 to 3 heteroatoms, of which R ⁵ and R ⁶ an atom attached to it to form a selectively substituted cyclic hydrocarbyl ring; R ² is a heteroatom functional group or a hydrocarbon group, the hydrocarbon group is selected from C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, and C6-C14 aryl, in which each alkyl, alkenyl and alkynyl are selectively cyclized, and each hydrocarbon group is selectively substituted and optionally contains 1 to 3 heteroatoms; R ³ is halogen, lower Alkyl, hydroxyl, lower alkoxy; wherein n is 2 or 3 substituents selected from halogen, -R ', - OR', = O, -NR'R ", - CO 2 R ' and -NR" CO ₂ R', wherein R ' And R "are each independently hydrogen, unsubstituted (C ₁ -C ₈ ) alkyl, or unsubstituted aryl. For example, the compound in the first scope of the patent application, wherein: R ¹ is -OR ⁴ or -NR ⁵ R ⁶ , or a C6-C14 aryl group, wherein the aryl group is optionally substituted and optionally contains 1 to 3 heterocyclic groups. Atom, and each of R ⁴ , R ^5, and R ^{6 is} independently H or a hydrocarbon group, the hydrocarbon group is selected from C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, and C6-C14 aryl, wherein each Alkyl, alkenyl, and alkynyl systems are selectively cyclized, and each hydrocarbon group is selectively substituted and optionally contains 1 to 3 heteroatoms, of which R ⁵ and R ⁶ are atoms connected to them, each A selectively substituted cyclic hydrocarbyl ring can be formed. For example, the compound in the scope of patent application, wherein R ¹ is -NR ⁵ R ⁶ , and R ⁵ and R ^{6 are} each independently H or a hydrocarbon group, and the hydrocarbon group is selected from C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, and C6-C14 aryl, wherein each alkyl, alkenyl, and alkynyl system is selectively cyclized, and each hydrocarbyl system is selectively substituted, and optionally contains 1 to 3 heteroatoms . For example, the compound of claim 1 in which R ¹ is -NH ₂ . For example, the compound of claim 1 in which R ² is -OR ⁴ or -NR ⁵ R ⁶ , or a hydrocarbon group, and the hydrocarbon group is selected from C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, and C6-C14 aryl, in which each alkyl, alkenyl and alkynyl system is selectively cyclized, and each hydrocarbon group is selectively substituted and optionally contains 1 to 3 heteroatoms, among which R ⁴ , R ⁵ and R ^{6 are} each independently H or a hydrocarbon group, and the hydrocarbon group is selected from C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, and C6-C14 aryl, wherein each alkyl, alkenyl and alkynyl The radical is selectively cyclized, and each hydrocarbon group is selectively substituted, and optionally contains 1 to 3 heteroatoms. Among them, the atoms to which R ⁵ and R ⁶ are connected together can each form a selectively substituted Cycloalkyl ring. For example, the compound in the first scope of the patent application, wherein: R ² is -NR ⁵ R ⁶ or a hydrocarbon group, and the hydrocarbon group is selected from C3-C8 cycloalkyl, C5-C8 cycloalkenyl, and C6-C14 aryl, wherein each hydrocarbon group Is optionally substituted, and optionally contains 1 to 3 heteroatoms, wherein R ⁴ , R ⁵ and R ^{6 are} each independently H or a hydrocarbon group, the hydrocarbon group is selected from C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, and C6-C14 aryl, wherein each alkyl, alkenyl, and alkynyl system is selectively cyclized, and each hydrocarbon group is selectively substituted, and optionally contains 1 to 3 heteroatoms Among them, the atoms to which R ⁵ and R ⁶ are connected together may each form a selectively substituted cyclic hydrocarbon ring. For example, the compound of the scope of patent application, wherein: R ² is -NR ⁵ R ⁶ or C6-C14 aryl, wherein aryl is optionally substituted, and optionally contains 1 to 3 heteroatoms, and R ⁴ , R ⁵ and R ^{6 are} each independently H or a hydrocarbon group, and the hydrocarbon group is selected from C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, and C6-C14 aryl, wherein each alkyl, olefin And alkynyl are selectively cyclized, and each hydrocarbyl is selectively substituted, and optionally contains 1 to 3 heteroatoms, wherein the atoms to which R ⁵ and R ⁶ are connected together can each form a selectivity A substituted cyclic hydrocarbyl ring. For example, the compound in the scope of patent application, or the pharmaceutically acceptable salt thereof, is selected from: A medicinal composition comprising a therapeutically effective amount of a compound according to any one of claims 1 to 8 in a unit dosage form or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers . A combination comprising a therapeutically effective amount of a compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof and a different agent having anticancer therapeutic activity.