Biphenyl derivatives, pharmaceutical compositions and their use

Abstract

This application relates to biphenyl derivatives, wherein the biphenyl derivatives have the structure represented by formula I, or are pharmaceutically acceptable salts, esters, amides, solvates, active metabolites, polymorphs, isotope-labeled, isomers, or prodrugs of the structure represented by formula I. Here, ring A is a methyl- or amino-substituted, or unsubstituted, five-membered ring, and the ring atoms of ring A contain one or two nitrogen atoms, R 1 It is a hydrogen bond donor or acceptor, and its structure contains one or more of the following: an amino group, an imino group, a hydroxyl group, and an ether bond. JPEG2026528760000075.jpg6154

Description

[Technical Field]

[0001] This application relates to the pharmaceutical technology field, and more particularly to biphenyl derivatives, pharmaceutical compositions, and their use. [Background technology]

[0002] In 2006, Shinya Yamanaka's research team proposed a so-called "cocktail" method consisting of four transcription factors: Oct4, Sox2, KlF4, and c-Myc, and succeeded in reprogramming terminally differentiated skin fibroblasts into pluripotent stem cells.

[0003] Such stem cells are called induced pluripotent stem cells (Takahashi K, et al., Cell, 2006, 126(4) pp.663-676; Takahashi K and Yamanaka S, Cell, 2007, 131(5) pp.861-872). These stem cells have differentiation potential similar to embryonic stem cells, forming the three most basic germ layers in human development: ectoderm, mesoderm, and endoderm, and ultimately forming a variety of adult cells. The proposal of this method overcomes the ethical constraints associated with the use of human embryonic stem cells in the medical field and greatly expands the potential of stem cell technology to be applied to clinical medicine.

[0004] On the one hand, many of the currently widely used reprogramming methods use viruses or other types of vectors to overexpress reprogramming transcription factors represented by Oct4 (Takahashi K, et al., Cell, 2006, 126(4):663 - 676; Takahashi K and Yamanaka S, Cell, 2007, 131(5):861 - 872; Yu J, et al., Science. 2007;318:1917 - 1920). However, such methods have potential clinical risks in the clinical application of induced pluripotent stem cells (iPSCs), such as the risk of tumor formation caused by the use of viral vectors. In addition, the complex GMP manufacturing process dependent on vectors leads to the complication of the clinical regulation of induced pluripotent stem cells, and there is also the problem that the manufacturing cost increases due to the use of vectors. Therefore, if selective gene regulation can be achieved using chemical substances and the expression state of reprogramming transcription factors can be changed, reprogramming that does not require the ectopic expression of "reprogramming genes" becomes possible.

[0005] The Wnt / β - catenin signaling pathway is an evolutionarily conserved signaling cascade that is extremely important in embryonic development, cell activity, and tissue regeneration, and its dysregulation is also associated with tumor formation (H. Clevers and R. Nusse, Cell, 2012, 149:1192 - 1205). Therefore, the regulation of the upstream and downstream of the Wnt signaling pathway has attracted attention in multiple research fields.

[0006] Currently, several core protein inhibitors in the Wnt signaling pathway are being developed and applied. GSK3 is a serine / threonine kinase and an important inhibitor in the Wnt pathway. CHIR99021 is a widely used GSK3 inhibitor and has been reported to have a remarkable effect in maintaining pluripotency in mammalian embryos (Meek et al., STEM CELLS, 2007, 31, 10, pp. 2104-2115). Furthermore, it has been shown that by using CHIR99021 in combination with compounds such as valproic acid, fibroblasts can be converted into pluripotent stem cells through pure chemical reprogramming (without using genetic factors) (Guan et al., Nature, 2022 May; 605, 790, : 325-331). Recent studies have shown that the use of CHIR99021 enhances cellular pluripotency by causing GSK3-downstream β-catenin to form a complex with Oct4 in a TCF-independent manner (Fernando F et al., Development, 2013:140, 1171-1183).

[0007] The CMGC kinase family is named after the initial letters of its subfamily names, which include cell cycle-dependent kinases (CDKs), mitogen-activated protein kinases (MAPKs), glycogen synthase kinases (GSKs), and CDC-like kinases (CLKs). CMGC kinases are highly conserved in living organisms (Chowdhury I et al., Cancers (Basel), 2023 Jul 28;15(15):3838). For example, CLK has the ability to phosphorylate serine, threonine, and tyrosine residues (Manning G et al., Science, 2002;298:1912-1934). CLK has a very typical structure, consisting of an N-lobe and a C-lobe, which are linked by a "hinge" region of the protein backbone. The structure distributed between the N- and C-regions is a catalytic domain consisting of a β-chain and an α-helix (Bullock A et al., Structure, 2009;17:352-362; Keri G et al., Curr. Signal Transduct. Ther. 2006;1:67-95). CLK is involved in regulating the Wnt signaling pathway by adjusting the alternative splicing of major Wnt-related genes (Tam BY et al., Cancer Lett. 2019:S0304383519304732). Representative examples of CLK kinases include CLK3 and CLK4. Another subfamily of CMGCs, mitogen-activated protein kinases (MAPKs), are serine-threonine protein kinases that are activated by various extracellular stimuli such as cytokines, neurotransmitters, hormones, cell stress, and cell adhesion. A representative example of this group is JNK1 kinase (c-Jun N-terminal kinase 1).

[0008] The STE kinase family is involved in the regulation of the MAP kinase and CMGC kinase families and plays a central role in various extracellular and intracellular signal transduction. For example, hPAK1, a member of the STE20 kinase family, controls the JNK MAP kinase pathway via GTPase (Brown J.L. et al., Curr Biol., 1996 May 1;6(5):598-605). TNIK (Traf2 and Nck-interacting kinase), a member of the STE kinase family, is also one of the regulators of the β-catenin-TCF4 transcription complex. TNIK promotes the activation of downstream target genes of the Wnt signal transduction pathway by directly interacting with β-catenin and TCF4 and plays an important role in cytoskeleton formation and development processes (Fu et al., JBC, 1999, (274):30729-30737; Mahmoudi et al., EMBO, 2009, (28):3329-3340). The TNIK inhibitor NCB-0846 has been reported to negatively regulate the TGF-β / SMAD signal transduction pathway by reducing the expression of TGFBR1 and inhibit the epithelial-mesenchymal transition process in tumors (Sugano et al., Br J Cancer, 2021, (124):228-236).

[0009] In addition to the core proteins of the Wnt pathway, it has now been shown that several kinases located upstream and downstream of the Wnt pathway are important regulators of Wnt signaling. For example, tyrosine kinases are enzymes that catalyze the transfer of phosphate groups from ATP to tyrosine residues of proteins in cells, and play a role in regulating the "on" and "off" states of intracellular signaling pathways. Receptor tyrosine kinases (RTKs) are a type of tyrosine kinase that is activated when a ligand binds to its extracellular domain (Hanks SK et al., Science, 1988 Jul 1;241(4861):42-52). PDGFRs are one of the core members of the receptor tyrosine kinase (RTK) family and include two subtypes: PDGFRα and PDGFRβ (Guerit et al., Cellular and Molecular Life Sciences, 2021, 78:3867-3881). PDGFRα can promote the early development of multiple cell lineages by inhibiting the Wnt9a / β-catenin signaling pathway (Bartoletti et al., Developmental Biology, 2020, (1):36-46; Sun et al., Cell Stem Cell, 2020(26):707-721). SU5402 is an inhibitor that acts on all of the FGF, VEGF, and PDGF signaling pathways, and regulates Wnt signaling by acting on platelet-derived growth factor receptors (PDGFRs).

[0010] Notably, while both the upstream and downstream pathways of the Wnt pathway mentioned above have some influence on the Wnt signaling pathway, none of the GSK3 inhibitor CHIR99021, the PDGF signaling pathway inhibitor SU5402, and the TNIK inhibitor NCB-0846 can control specific physiological changes. However, none of them can simultaneously inhibit multiple Wnt pathway upstream kinases. Furthermore, they cannot increase the expression of OCT4, a reprogramming core gene, during the somatic cell reprogramming process, thereby influencing other reprogramming genes and increasing the probability of somatic cell reprogramming occurring. [Overview of the Initiative] [Problems that the invention aims to solve]

[0011] Based on the above, there is a need to provide biphenyl derivatives and pharmaceutical compositions containing them. These derivatives are designed based on multiple structurally similar Wnt upstream kinases, can control at least one Wnt upstream pathway, and can increase the expression of the reprogramming core genes Oct4, Lin28A, and c-Myc even when present alone. Therefore, they can be well applied to cell reprogramming. [Means for solving the problem]

[0012] A first aspect of this application provides a biphenyl derivative, which has the structure represented by formula I, or is a pharmaceutically acceptable salt, ester, amide, solvate, active metabolite, polymorph, isotope-labeled, isomer, or prodrug of the structure represented by formula I. [ka] (In the formula, ring A is a methyl group or amino group substituted or unsubstituted five-membered ring, and the ring atoms of ring A contain one or two nitrogen atoms.) R 1(A hydrogen bond is a hydrogen bond donor or acceptor, and its structure contains one or more of the following: an amino group, an imino group, a hydroxyl group, and an ether bond.)

[0013] A second aspect of this application provides a pharmaceutical composition comprising the aforementioned biphenyl derivative and at least one pharmaceutically acceptable carrier.

[0014] A third aspect of this application provides the application of the aforementioned biphenyl derivative or pharmaceutical composition in cell reprogramming.

[0015] A fourth aspect of this application provides a method for cell reprogramming, The process includes the step of treating cells by contacting them with the aforementioned biphenyl derivative or pharmaceutical composition.

[0016] A fifth aspect of this application is the application of the aforementioned biphenyl derivative or pharmaceutical composition in kinase inhibition, wherein the kinase comprises one or more of the TK kinase family, the STE kinase family and the CMGC kinase family.

[0017] A sixth aspect of this application provides a method for kinase inhibition, The process includes the step of contacting the kinase with the aforementioned biphenyl derivative or pharmaceutical composition, The kinase comprises one or more of the TK kinase family, the STE kinase family, and the CMGC kinase family. [Brief explanation of the drawing]

[0018] [Figure 1]This shows a comparative analysis of the three-dimensional structures of PDGFR, TNIK, CLK4, and JNK1 proteins. Figure 1A is a heatmap display of structural similarity (RMSD) obtained from a bimodal comparison of protein structures, and the numerical values ​​in the figure represent the RMSD of the bimodal structural comparison. Figure 1B, Figure 1C, Figure 1D, and Figure 1E are PDGFR (PDB ID: 5GRN) (Liang L et al., Biochem Biophys Res Commun.2016 Sep 2;477(4):667-672), TNIK (PDB ID: 5AX9) (Masuda M et al., Nat Commun, 2016 Aug), respectively. 26;7:12586), CLK4 (PDB ID:6FYV) (Kallen J et al.,ChemMedChem.2018 Sep 19;13(18):1997-2007) and JNK1 (PDB ID:3ELJ) (Chamberlain SD et al.,Bioorg Med Chem Lett.2009 Jan This is a cartoon model of the three-dimensional structure of 15;19(2):360-4), with the important sites of the protein's active site (Figure 1B) shown as stick models. [Figure 2]Molecular dynamics simulations demonstrate the binding ability and chemical bonding details between the biphenyl derivative I-1 and TNIK. Figure A shows the change in RMSD of the TNIK-I-1 complex over simulation time during the simulation process. The results indicate that TNIK binds to I-1 and that the complex structure formed by the binding is in a stable binding state. Figure B is an energy decomposition diagram of the interaction between each amino acid site of TNIK and I-1. The x-axis represents the amino acid sequence of TNIK, and the y-axis represents the energy value that each amino acid site contributes to ligand binding. The names of amino acid sites with an energy contribution to ligand binding of less than -0.5 kJ / mol are shown. Figure C shows the complex structure of TNIK-I-1 after the molecular dynamics simulation has stabilized. The protein is displayed as a cartoon model, the color of each site indicates the energy contribution to ligand binding, the small molecule and the main chain or side chain of the residues interacting with it are displayed as stick models, hydrogen bonding interactions are shown by long dashed lines, and hydrophobic interactions are shown by short dashed lines. These results clearly demonstrate the chemical basis of the active catalytic center of the kinase target to which TNIK and I-1 bind, and the interaction between them, indicating that the basis for chemical bond formation is consistent. [Figure 3]Molecular dynamics simulations demonstrate the binding ability and chemical bonding details between the biphenyl derivative I-4 and TNIK. Figure A shows the change in RMSD of the TNIK-I-1 complex over simulation time during the simulation process. The results indicate that TNIK binds to I-4 and that the complex structure formed by the binding is in a stable binding state. Figure B is an energy decomposition diagram of the interaction between each amino acid site of TNIK and I-4. The x-axis represents the amino acid sequence of TNIK, and the y-axis represents the energy value that each amino acid site contributes to ligand binding. The names of amino acid sites with an energy contribution to ligand binding of less than -0.5 kJ / mol are shown. Figure C shows the complex structure of TNIK-I-4 after the molecular dynamics simulation has stabilized. The protein is displayed as a cartoon model, the color of each site indicates the energy value that contributes to ligand binding, the small molecule and the main chain or side chain of the residues interacting with it are displayed as stick models, hydrogen bonding interactions are shown by long dashed lines, and hydrophobic interactions are shown by short dashed lines. These results clearly demonstrate the chemical basis of the active catalytic center of the kinase target to which TNIK and I-4 bind, and the interaction between them, indicating that the basis for chemical bond formation is consistent. [Figure 4]Molecular dynamics simulations demonstrate the binding ability and chemical bonding details between the biphenyl derivative I-12 and TNIK. Figure A shows the change in RMSD of the TNIK-I-12 complex over simulation time during the simulation process. The results indicate that TNIK binds to I-12 and that the complex structure formed by the binding is in a stable binding state. Figure B is an energy decomposition diagram of the interaction between each amino acid site of TNIK and I-12. The x-axis represents the amino acid sequence of TNIK, and the y-axis represents the energy value that each amino acid site contributes to ligand binding. The names of amino acid sites with an energy contribution to ligand binding of less than -0.5 kJ / mol are shown. Figure C shows the complex structure of TNIK-I-12 after the molecular dynamics simulation has stabilized. The protein is displayed as a cartoon model, the color of each site indicates the energy value that contributes to ligand binding, the small molecule and the main chain or side chain of the residues interacting with it are displayed as stick models, hydrogen bonding interactions are shown by long dashed lines, and hydrophobic interactions are shown by short dashed lines. These results clearly demonstrate the chemical basis of the active catalytic center of the kinase target to which TNIK and I-12 bind, and the interaction between them, indicating that the basis for chemical bond formation is consistent. [Figure 5]Molecular dynamics simulations demonstrate the binding ability and chemical bonding details between the biphenyl derivative I-1 and PDGFRα. Figure A shows the change in RMSD of the PDGFRα-I-1 complex over simulation time during the simulation process. The results indicate that PDGFRα binds to I-1 and that the complex structure formed by the binding is in a stable binding state. Figure B is an energy decomposition diagram of the interaction between each amino acid site of PDGFRα and I-1. The x-axis represents the amino acid sequence of PDGFRα, and the y-axis represents the energy value that each amino acid site contributes to ligand binding. The names of amino acid sites with an energy contribution to ligand binding of less than -0.5 kJ / mol are shown. Figure C shows the complex structure of PDGFRα-I-1 after the molecular dynamics simulation has stabilized. The protein is displayed as a cartoon model, the color of each site indicates the energy value that contributes to ligand binding, the small molecule and the main chain or side chain of the residues interacting with it are displayed as stick models, hydrogen bonding interactions are shown by long dashed lines, and hydrophobic interactions are shown by short dashed lines. These results clearly demonstrate the chemical basis of the active catalytic center of the kinase target to which PDGFRα and I-1 bind, and the interaction between them, indicating that the basis for chemical bond formation is consistent. [Figure 6]Molecular dynamics simulations demonstrate the binding ability and chemical bonding details between the biphenyl derivative I-4 and PDGFRα. Figure A shows the change in RMSD of the PDGFRα-I-1 complex over simulation time during the simulation process. The results indicate that PDGFRα binds to I-1 and that the complex structure formed by the binding is in a stable binding state. Figure B is an energy decomposition diagram of the interaction between each amino acid site of PDGFRα and I-4. The x-axis represents the amino acid sequence of PDGFRα, and the y-axis represents the energy value that each amino acid site contributes to ligand binding. The names of amino acid sites with an energy contribution to ligand binding of less than -0.5 kJ / mol are shown. Figure C shows the complex structure of PDGFRα-I-4 after the molecular dynamics simulation has stabilized. The protein is displayed as a cartoon model, the color of each site indicates the energy contribution to ligand binding, the small molecule and the main chain or side chain of the residues interacting with it are displayed as stick models, hydrogen bonding interactions are shown by long dashed lines, and hydrophobic interactions are shown by short dashed lines. These results clearly demonstrate the chemical basis of the active catalytic center of the kinase target to which PDGFRα and I-4 bind, and the interaction between them, indicating that the basis for chemical bond formation is consistent. [Figure 7]Molecular dynamics simulations demonstrate the binding ability and chemical bonding details between the biphenyl derivative I-12 and PDGFRα. Figure A shows the change in RMSD of the PDGFRα-I-12 complex over simulation time during the simulation process. The results indicate that PDGFRα binds to I-12, and that the complex structure formed by this binding is in a stable binding state. Figure B is an energy decomposition diagram of the interaction between each amino acid site of PDGFRα and I-12. The x-axis represents the amino acid sequence of PDGFRα, and the y-axis represents the energy value that each amino acid site contributes to ligand binding. The names of amino acid sites with an energy contribution to ligand binding of less than -0.5 kJ / mol are shown. C shows the complex structure of PDGFRα and I-12 after molecular dynamics simulations have stabilized. The protein is shown as a cartoon model, with the color of each site indicating the energy contribution to ligand binding. Small molecules and the main chains or side chains of the residues interacting with them are shown as stick models, hydrogen bonding interactions are indicated by long dashed lines, and hydrophobic interactions are indicated by short dashed lines. This result clearly shows the chemical basis of the active catalytic center of the kinase target to which PDGFRα and I-12 bind and their interactions, i.e., the basis for chemical bond formation is consistent. [Figure 8]Molecular dynamics simulations demonstrate the binding ability and chemical bonding details between the biphenyl derivative I-1 and CLK4. Figure A shows the change in RMSD of the CLK4-I-1 complex over simulation time during the simulation process. The results indicate that CLK4 binds to I-1 and that the complex structure formed by the binding is in a stable binding state. Figure B is an energy decomposition diagram of the interaction between each amino acid site of CLK4 and I-1. The x-axis represents the amino acid sequence of CLK4, and the y-axis represents the energy value that each amino acid site contributes to ligand binding. The names of amino acid sites with an energy contribution to ligand binding of less than -0.5 kJ / mol are shown. Figure C shows the complex structure of CLK4-I-1 after the molecular dynamics simulation has stabilized. The protein is displayed as a cartoon model, the color of each site indicates the energy contribution to ligand binding, the small molecule and the main chain or side chain of the residues interacting with it are displayed as stick models, hydrogen bonding interactions are shown by long dashed lines, and hydrophobic interactions are shown by short dashed lines. These results clearly demonstrate the chemical basis of the active catalytic center of the kinase target to which CLK4 and I-1 bind, and the interaction between them, indicating that the basis for chemical bond formation is consistent. [Figure 9]Molecular dynamics simulations demonstrate the binding ability and chemical bonding details between the biphenyl derivative I-4 and CLK4. Figure A shows the change in RMSD of the CLK4-I-1 complex over simulation time during the simulation process. The results indicate that CLK4 binds to I-4 and that the complex structure formed by the binding is in a stable binding state. Figure B is an energy decomposition diagram of the interaction between each amino acid site of CLK4 and I-4. The x-axis represents the amino acid sequence of CLK4, and the y-axis represents the energy value that each amino acid site contributes to ligand binding. The names of amino acid sites with an energy contribution to ligand binding of less than -0.5 kJ / mol are shown. Figure C shows the complex structure of CLK4-I-4 after the molecular dynamics simulation has stabilized. The protein is displayed as a cartoon model, the color of each site indicates the energy value that contributes to ligand binding, the small molecule and the main chain or side chain of the residues interacting with it are displayed as stick models, hydrogen bonding interactions are shown by long dashed lines, and hydrophobic interactions are shown by short dashed lines. These results clearly demonstrate the chemical basis of the active catalytic center of the kinase target to which CLK4 and I-4 bind, and the interaction between them, indicating that the basis for chemical bond formation is consistent. [Figure 10]Molecular dynamics simulations demonstrate the binding ability and chemical bonding details between the biphenyl derivative I-12 and CLK4. Figure A shows the change in RMSD of the CLK4-I-1 complex over simulation time during the simulation process. The results indicate that CLK4 binds to I-12 and that the complex structure formed by the binding is in a stable binding state. Figure B is an energy decomposition diagram of the interaction between each amino acid site of CLK4 and I-12. The x-axis represents the amino acid sequence of CLK4, and the y-axis represents the energy value that each amino acid site contributes to ligand binding. The names of amino acid sites with an energy contribution to ligand binding of less than -0.5 kJ / mol are shown. Figure C shows the complex structure of CLK4-I-12 after the molecular dynamics simulation has stabilized. The protein is displayed as a cartoon model, the color of each site indicates the energy value that contributes to ligand binding, the small molecule and the main chain or side chain of the residues interacting with it are displayed as stick models, hydrogen bonding interactions are shown by long dashed lines, and hydrophobic interactions are shown by short dashed lines. These results clearly demonstrate the chemical basis of the active catalytic center of the kinase target to which CLK4 and I-12 bind, and the interaction between them, indicating that the basis for chemical bond formation is consistent. [Figure 11]Molecular dynamics simulations demonstrate the binding ability and chemical bonding details between biphenyl derivative I-1 and JNK1. Figure A shows the change in RMSD of the JNK1-I-1 complex over simulation time during the simulation process. The results indicate that JNK1 binds to I-1 and that the complex structure formed by the binding is in a stable binding state. Figure B is an energy decomposition diagram of the interaction between each amino acid site of JNK1 and I-1. The x-axis represents the amino acid sequence of JNK1, and the y-axis represents the energy value that each amino acid site contributes to ligand binding. The names of amino acid sites with an energy contribution to ligand binding of less than -0.5 kJ / mol are shown. Figure C shows the complex structure of JNK1-I-1 after the molecular dynamics simulation has stabilized. The protein is displayed as a cartoon model, the color of each site indicates the energy value that contributes to ligand binding, the small molecule and the main chain or side chain of the residues interacting with it are displayed as stick models, hydrogen bonding interactions are shown by long dashed lines, and hydrophobic interactions are shown by short dashed lines. These results clearly demonstrate the chemical basis of the active catalytic center of the kinase target to which JNK1 and I-1 bind, and the interaction between them, indicating that the basis for chemical bond formation is consistent. [Figure 12]Molecular dynamics simulations demonstrate the binding ability and chemical bonding details between the biphenyl derivative I-4 and JNK1. Figure A shows the change in RMSD of the JNK1-I-4 complex over simulation time during the simulation process. The results indicate that JNK1 binds to I-4 and that the complex structure formed by the binding is in a stable binding state. Figure B is an energy decomposition diagram of the interaction between each amino acid site of JNK1 and I-4. The x-axis represents the amino acid sequence of JNK1, and the y-axis represents the energy value that each amino acid site contributes to ligand binding. The names of amino acid sites with an energy contribution to ligand binding of less than -0.5 kJ / mol are shown. Figure C shows the complex structure of JNK1-I-4 after the molecular dynamics simulation has stabilized. The protein is displayed as a cartoon model, the color of each site indicates the energy contribution to ligand binding, the small molecule and the main chain or side chain of the residues interacting with it are displayed as stick models, hydrogen bonding interactions are shown by long dashed lines, and hydrophobic interactions are shown by short dashed lines. These results clearly demonstrate the chemical basis of the active catalytic center of the kinase target to which JNK1 and I-4 bind, and the interaction between them, indicating that the basis for chemical bond formation is consistent. [Figure 13] This shows a visualization analysis of the activity assay results of the biphenyl derivative I-1 across a kinase panel range. Here, each node represents a single kinase, and both the size and color intensity of the nodes indicate the inhibitory effect of I-1 on each kinase. The results demonstrate that I-1 has actual binding activity against each target kinase. The phylogenetic tree of the kinase panel was referenced from Cell Signaling Technology (www.cellsignal.com). [Figure 14] Based on kinase activity screening analysis, the kinase targets on which biphenyl derivatives act are shown. The effects of biphenyl derivative I-1 on the enzymatic activity of 330 kinases are shown (the x-axis displays one kinase name for every 10 kinases). [Figure 15]The IC50 results for biphenyl derivatives against target kinases are shown, and the data indicate that biphenyl derivatives not only exhibit binding ability to target kinases but also possess significant antagonistic activity against them. [Figure 16] This study demonstrates that biphenyl derivatives significantly enhance the expression of the reprogramming core genes OCT4, c-Myc, and Lin28A. Compared to a blank control (CK) without any added compounds, biphenyl derivatives, even when present alone, can simultaneously enhance the expression of this group of reprogramming core genes. [Modes for carrying out the invention]

[0019] To facilitate understanding of this application, a more comprehensive description of this application will be provided below with reference to the relevant drawings. While the drawings illustrate preferred embodiments of this application, it can be implemented in a variety of forms and is not limited to the embodiments described herein. Rather, the purpose of presenting these embodiments is to provide a more complete and comprehensive understanding of the disclosures of this application.

[0020] Unless otherwise specified, all technical and scientific terms used herein have meanings that are generally understood by those skilled in the art. The terms used herein are for illustrative purposes of specific embodiments and are not intended to limit this application. The terms “and / or” used herein encompass any one or more of the listed items, or any combination thereof.

[0021] In this application, the disclosed technical features include confidential technical solutions consisting of the listed features, as well as open technical solutions that include the listed features.

[0022] In this application, unless otherwise specified, numerical ranges are understood as continuous ranges, including the minimum and maximum values ​​of the range, and all values ​​between the minimum and maximum values. Furthermore, if a range refers to an integer, it includes all integers between the minimum and maximum values ​​of that range. In addition, for features or characteristics to which multiple ranges are assigned, these ranges can be combined. In other words, unless otherwise specified, all ranges disclosed herein are understood to include any and all subranges contained therein.

[0023] In this application, unless otherwise specified, percentage content refers to mass percentage for solid-liquid mixtures and solid-phase-solid mixtures, and volume percentage for liquid-phase-liquid mixtures.

[0024] In this application, unless otherwise specified, percentage concentrations refer to the final concentration. The final concentration, as used herein, means the proportion of the component in the system after its addition.

[0025] Unless otherwise specified, the temperature parameters in this application may be constant temperature processing or processing within a specific temperature range. Constant temperature processing, as used herein, means that temperature fluctuations are permitted within a precision range controlled by the equipment.

[0026] (Explanation of terms) In this specification, "ring atom" refers to an atom involved in the formation of the A ring skeleton and does not include substituents on the A ring. For example, if the A ring has the structure represented by A-1-1, the ring atoms are the carbon atoms of the two bonding sites, the two nitrogen atoms, and the carbon atom that forms the carbon-nitrogen double bond, and the carbon atom in the substituted methyl group on the nitrogen atom does not belong to the ring atoms.

[0027] A "hydrogen bond donor" refers to a group containing a hydrogen atom bonded to an atom with high electronegativity and a small radius, such as oxygen, nitrogen, or fluorine, and includes groups such as amino groups and hydroxyl groups. A "hydrogen bond acceptor" refers to a group containing an atom with high electronegativity and a small radius, such as oxygen, nitrogen, or fluorine. In this application, R 1 It may be a hydrogen bond donor or acceptor, and inhibits kinase by forming hydrogen bonds with the protein structure.

[0028] In this specification, “prodrug” refers to a compound that, upon administration to a living organism, produces a drug, i.e., an active ingredient, through spontaneous chemical reactions, enzymatic catalyzed chemical reactions, photolysis, and / or metabolic chemical reactions. Therefore, a prodrug is a covalently modified analog or potential form of a therapeutically active compound. Suitable examples include, but are not limited to, carboxylic acid esters, carbonate esters, phosphate esters, nitrate esters, sulfate esters, sulfone esters, sulfinate esters, amino compounds, carbamates, azo compounds, phosphoramides, glucosides, ethers, and acetals of compounds.

[0029] "Medically acceptable" means a ligand, material, composition, and / or dosage form that is appropriate for administration to a patient within the bounds of reasonable medical judgment and that has a reasonable benefit-to-risk ratio.

[0030] "Medically acceptable carrier" refers to a pharmaceutically acceptable material, composition, or medium, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. As used herein, the term "medically acceptable carrier" includes buffers suitable for drug administration, sterile water for injection, solvents, dispersions, coatings, antimicrobial and antifungal agents, isotonic agents, and absorption retarders. Each carrier must be "medically acceptable" in the sense of compatibility with other components in the formulation and non-harmful to the patient. Appropriate examples include: (1) sugars such as lactose, glucose, and sucrose; (2) starches such as corn starch, potato starch, and substituted or unsubstituted β-cyclodextrins; (3) cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, and acetylcellulose; (4) powdered tragacanth gum; (5) malt; (6) gelatin; (7) talc; (8) excipients such as cocoa butter and suppository waxes; (9) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil. (10) Glycols such as propylene glycol, (11) Polyols such as glycerol, sorbitol, mannitol, and polyethylene glycol, (12) Esters such as ethyl oleate and ethyl laurate, (13) Agar, (14) Buffers such as magnesium hydroxide and aluminum hydroxide, (15) Alginic acid, (16) Pyrogen-free water, (17) Isotonic saline, (18) Ringer's solution, (19) Ethanol, (20) Phosphate buffer, (21) Other non-toxic, suitable substances used in pharmaceutical formulations.

[0031] A "pharmaceutically acceptable salt" refers to a salt formed by any compound having the given structure with an acid or base suitable for use as a pharmaceutical. pharmaceutically acceptable salts include both inorganic and organic salts. Class I salts are salts formed by the compounds of this application with acids. Acids suitable for salt formation include, but are not limited to, inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, and phosphoric acid; organic acids such as formic acid, acetic acid, trifluoroacetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, benzoic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, and naphthalenesulfonic acid; and amino acids such as proline, phenylalanine, aspartic acid, and glutamic acid. Another Class I salts are salts formed by the compounds of this application with bases. Suitable bases for salt formation include, but are not limited to, alkali metal salts (e.g., sodium or potassium salts), alkaline earth metal salts (e.g., magnesium or calcium salts), ammonium salts (e.g., lower alkanolammonium salts and other pharmaceutically acceptable amine salts), such as methylamine salts, ethylamine salts, propylamine salts, dimethylamine salts, trimethylamine salts, diethylamine salts, triethylamine salts, tert-butylamine salts, ethylenediamine salts, hydroxyethylamine salts, dihydroxyethylamine salts, trihydroxyethylamine salts, and amine salts formed from morpholine, piperazine, and lysine, respectively.

[0032] "Medically acceptable esters and amides" refers to esters or amides suitable for pharmaceutical use, formed by any of the compounds having the indicated structure in combination with other compounds. Medicinally acceptable esters include organic esters and inorganic esters.

[0033] Similarly, if the compound represented by formula I has a hydroxyl group, a pharmaceutically acceptable ester can be obtained by condensing the compound represented by formula I with a carboxylic acid, acyl chloride, acid anhydride, etc., according to a standard method.

[0034] The aforementioned esters include, for example, C1-C6 alkyl esters such as methyl esters, ethyl esters, propyl esters, isopropyl esters, butyl esters, sec-butyl esters, tert-butyl esters, pentyl esters, and hexyl esters; C3-C6 cycloalkyl esters such as cyclopentyl esters and cyclohexyl esters; and C6-C alkyl esters such as phenyl esters and naphthyl esters. 10 Aryl esters; such as benzyl esters, phenethyl esters, α-methylbenzyl esters, 3-phenylpropyl esters, 4-phenylbutyl esters, 6-phenylhexyl esters, diphenylmethyl esters, triphenylmethyl esters, etc. C6-C 10 Examples include aryl C1-C6 alkyl esters. Alternatively, they may be esters that can be hydrolyzed in vivo, such as (5-methyl-2-oxo-1,3-dioxacyclopenten-4-yl)methyl ester, (pivaloyloxy)methyl ester, benzofuranone ester, [(isopropoxycarbonyl)oxy]methyl ester, [(cyclohexyloxycarbonyl)oxy]methyl ester, and 1-[(cyclohexyloxycarbonyl)oxy]ethyl ester.

[0035] The amide includes, for example, mono-C1-C6 alkyl amides or mono-C3-C6 cycloalkyl amides such as amide (-CONH2), N-formamide, N-acetamide, N-propionamide, N-isopropionamide, N-butylamide, N-sec-butylamide, N-tert-butylamide, N-pentanamide, N-hexanamide, N-cyclopropionamide, N-cyclopentanamide, and N-cyclohexanamide. Or amides such as N,N-diformamide, N,N-diacetamide, N,N-dipropionamide, N,N-diisopropionamide, N-methyl-N-acetamide, N-methyl-N-propionamide, N-methyl-N-butylamide, N-ethyl-N-propionamide, N-ethyl-N-butylamide, N-butyl-N-cyclopentanamide, N-ethyl-N-cyclopropionamide, N,N-dicyclohexanamide are di-C1-C6 alkyl amides, N-C1-C6 alkyl-N-C3-C6 cycloalkyl amides, or di-C3-C6 cycloalkyl amides.

[0036] "Solvate" refers to a complex formed by the compound represented by the general formula (I) coordinating with solvent molecules in a specific ratio. "Hydrate" refers to a complex formed by the compound of the present application coordinating with water.

[0037] "Active metabolite" refers to a derivative having the activity of the compound, which is generated during the metabolism of the compound.

[0038] "Crystal polymorph" refers to the compound of the present application existing in different crystal lattice forms.

[0039] "Isotope labeling" refers to the compound of the present application labeled with isotopes. For example, as isotopes in the compound of the present application, various isotopes of elements such as H, C, N, O, P, F, S, for example, 2 H, 3 H, 13 C, 14 C, 15 N, 18 O, 17 O, 31 P, 32P, 35 S, 18 F, 36 This includes S, etc.

[0040] An "isomer" refers to an isomer resulting from a different spatial arrangement of atoms within a molecule. The compounds in this application contain structures such as chiral or asymmetric centers and double bonds, and therefore may include various isomeric forms such as optical isomers, geometric isomers, tautomers, and atropisomers. These isomers, their single isomers, and racemates are all included within the scope of this application. For example, optical isomers, specifically the optically active (R)-, (S)- isomers and D, L isomers, can be prepared by chiral separation, chiral synthesis, chiral reagents, or other conventional techniques. For instance, they can be converted to diastereomers by reacting with a suitable optically active substance (e.g., a chiral alcohol or Mosher's chloride), and then separated to convert them back to the corresponding single isomers (e.g., by hydrolysis). Furthermore, separation can also be performed by chromatography.

[0041] "Pharmaceutical compositions" can be prepared using methods well known in the pharmaceutical field and can be administered or used via various routes depending on whether local or systemic treatment is required and the area to be treated.

[0042] (Method of administration) There are no particular restrictions on the dosage form or method of use of the compound or pharmaceutical composition thereof of this application.

[0043] Typical administration methods include, but are not limited to, oral, intratumor, rectal, parenteral (intravenous, intramuscular, or subcutaneous) injection, and local administration.

[0044] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one common inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with any of the following components: (a) fillers or bulking agents such as starch, lactose, sucrose, glucose, mannitol, or silicic acid; (b) binders such as hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, or gum arabic; (c) humectants such as glycerin; (d) disintegrants such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, or sodium carbonate; (e) absorption retarders such as paraffin wax; (f) absorption enhancers such as quaternary ammonium compounds; (g) wetting agents such as cetyl alcohol or glyceryl monostearate; (h) adsorbents such as kaolin; or (i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In the form of capsules, tablets, and pills, buffering agents may be included in the dosage form. Solid dosage forms such as tablets, sugar-coated tablets, capsules, pills, and granules can be prepared using coating agents and shells, such as enteric coatings or other materials well known in the art. These may contain opacifying agents, and the release of the active compound or compound in such compositions may be delayed in a portion of the gastrointestinal tract. Examples of usable embedding components include polymers and waxes. If necessary, the active compound may also be prepared in microcapsule form using one or more of the above excipients.

[0045] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the active compound, the liquid dosage form may include inert diluents commonly used in the art, such as water or other solvents, solubilizers, emulsifiers, specifically, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, sesame oil, or mixtures thereof. In addition to these inert diluents, the composition may also include auxiliary agents such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents, and fragrances. For example, a suspension may include suspending agents, specifically, for example, ethoxylated isooctadecyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitan esters, microcrystalline cellulose, aluminum methoxide, and agar, or mixtures thereof.

[0046] Parenteral injection compositions may include physiologically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions, or emulsions, as well as sterile powders for redissolution in sterile injection solutions or dispersions. Suitable aqueous or non-aqueous carriers, diluents, solvents, or excipients include water, ethanol, polyols, and suitable mixtures thereof.

[0047] Topical dosage forms include ointments, powders, patches, sprays, and inhalants. These are prepared by mixing the active ingredient with a pharmaceutically acceptable carrier under sterile conditions, and any preservatives, buffers, or propellants as needed.

[0048] In this specification, “pharmaceutical” includes any drug, compound, composition, or mixture that produces physiological and / or pharmacological effects in or outside the body, and which often produce beneficial effects. The range of physiological and / or pharmacological effects that “pharmaceutical” produces in the body is not particularly limited and may include systemic effects or local effects only. The activity of “pharmaceutical” is not particularly limited and may include active substances that interact with other substances or inactive substances that do not interact with other substances.

[0049] A first aspect of this application provides biphenyl derivatives having the structure represented by formula I, or pharmaceutically acceptable salts, esters, amides, solvates, active metabolites, polymorphs, isotope-labeled, isomers, or prodrugs of the structure represented by formula I. [ka] (In the formula, ring A is a methyl group or amino group substituted or unsubstituted five-membered ring, and the ring atoms of ring A contain one or two nitrogen atoms.) R 1 (A hydrogen bond is a hydrogen bond donor or acceptor, and its structure contains one or more of the following: an amino group, an imino group, a hydroxyl group, and an ether bond.)

[0050] The biphenyl derivatives of this application, through structural design, involve condensing a five-membered ring containing one or two nitrogen atoms at the 2nd and 3rd positions of one benzene ring, and attaching a substituent capable of functioning as a hydrogen bond donor or acceptor at the 3' position of the other benzene ring. This allows for good binding to protein targets of kinases such as PDGFRα (belonging to the TK kinase family, more specifically the RTK kinase family), TNIK (belonging to the STE kinase family), CLK4 (belonging to the CMGC kinase family, more specifically the CLK kinase family), and JNK1 (belonging to the CMGC kinase family, more specifically the MAPK kinase family). As a result, the activity of these kinases can be simultaneously inhibited, thereby controlling the Wnt pathway from upstream. Furthermore, the expression of reprogramming core genes such as OCT4, Lin28A, and c-Myc can be significantly increased, effectively promoting cellular reprogramming. This demonstrates that a reprogramming effect can be achieved by simultaneously inhibiting multiple upstream kinases in the Wnt pathway.

[0051] In some embodiments, R 1 The following structure is selected: [ka] (The asterisk (*) indicates a connecting point.)

[0052] In some embodiments, R 1 The following structure is selected: [ka] (The asterisk (*) indicates a connecting point.)

[0053] Preferably, R 1 This is selected from R-1-1, R-1-2, R-2, or R-3-1.

[0054] In some embodiments, ring A is selected from one of the following structures. [ka] Here, X is CR 3 R 4 Or NR 5 And R 2 ~R 5 Each of these is independently selected from -H, -CH3, or -NH2. (The asterisk (*) indicates a connecting point.)

[0055] In some embodiments, ring A is selected from one of the following structures. [ka] (The asterisk (*) indicates a connecting point.)

[0056] Preferably, ring A is selected from one of the following structures. [ka] (The asterisk (*) indicates a connecting point.)

[0057] In some embodiments, the biphenyl derivative has a structure represented by any of formulas I-1 to I-18, or is a pharmaceutically acceptable salt, ester, amide, solvate, active metabolite, polymorph, isotope-labeled, isomer, or prodrug of any of the structures represented by formulas I-1 to I-18. [ka]

[0058] The inventors of this application selected 18 compounds represented by formulas I-1 to I-18 from among more than 1900 designed compounds, taking into account calculation results based on target proteins, kinase inhibitory effect, and difficulty of synthesis. Preferably, the compounds represented by formulas I-1 to I-4 and I-12 show better effects and are easy to synthesize.

[0059] A second aspect of this application provides a pharmaceutical composition comprising a biphenyl derivative according to one or more of the above embodiments and at least one pharmaceutically acceptable carrier.

[0060] A third aspect of this application provides applications of the biphenyl derivative or the pharmaceutical composition according to one or more embodiments in cell reprogramming.

[0061] In some embodiments, when a biphenyl derivative is used for cell reprogramming, the concentration of the biphenyl derivative is 1 μM to 50 μM. Optionally, the concentration of the biphenyl derivative may be, for example, 2 μM, 4 μM, 6 μM, 8 μM, 10 μM, 12 μM, 14 μM, 16 μM, 18 μM, 20 μM, 22 μM, 24 μM, 26 μM, 28 μM, 30 μM, 32 μM, 34 μM, 36 μM, 38 μM, 40 μM, 42 μM, 44 μM, 46 μM, or 48 μM.

[0062] A fourth aspect of this application provides a cell reprogramming method, which includes the following steps: The process includes the step of treating cells by contacting them with the biphenyl derivative or the pharmaceutical composition according to one or more embodiments.

[0063] In some embodiments, the concentration of the biphenyl derivative during the contact treatment is 1 μM to 50 μM. Optionally, the concentration of the biphenyl derivative during the contact treatment may be, for example, 2 μM, 4 μM, 6 μM, 8 μM, 10 μM, 12 μM, 14 μM, 16 μM, 18 μM, 20 μM, 22 μM, 24 μM, 26 μM, 28 μM, 30 μM, 32 μM, 34 μM, 36 μM, 38 μM, 40 μM, 42 μM, 44 μM, 46 μM, or 48 μM.

[0064] A fifth aspect of this application provides an application of the biphenyl derivative or pharmaceutical composition in kinase inhibition, wherein the kinase comprises one or more of the TK kinase family, the STE kinase family and the CMGC kinase family.

[0065] In some embodiments, the kinase comprises one or more of PDGFRα, TNIK, CLK4, and JNK1.

[0066] A sixth aspect of this application provides a method for kinase inhibition, the method comprising the following steps: The process includes the step of contacting the kinase with the biphenyl derivative or the pharmaceutical composition according to one or more embodiments, Kinases include one or more species from the TK kinase family, STE kinase family, and CMGC kinase family.

[0067] In some embodiments, the kinase comprises one or more of PDGFRα, TNIK, CLK4, and JNK1. Compounds I-1 to I-18 are synthesized via the following routes: (1) Synthesis of I-1 [ka] (S)-4-(3-(1-methyl-1H-indazole-4-yl)phenyl)pyrrolidine-2-one [ka]

[0068] To a solution of Pd(PPh3)4 (1.05 g, 0.91 mmol) in ethylene glycol dimethyl ether, a solution of (S)-4-(3-bromophenyl)pyrrolidine-2-one (4.51 g, 18.77 mmol) in ethylene glycol dimethyl ether (60 mL) was added and the mixture was stirred under a nitrogen atmosphere for 15 minutes. Next, a solution of 1-methyl-1H-indazole-4-boronic acid (5.00 g, 28.40 mmol) in ethanol (15 mL) was added to the mixture from the previous step and the mixture was stirred for 10 minutes. A 2 M aqueous solution of Na2CO3 (80 mL) was added to the resulting mixture, and it was dried under reflux for 20 hours. After the mixture cooled, the organic layer was separated, washed with brine, dried over sodium sulfate, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:benzene = 5:1) to obtain (S)-4-(3-(1-methyl-1H-indazole-4-yl)phenyl)pyrrolidine-2-one (5.25 g, 96%).

[0069] Liquid chromatography-mass spectrometry revealed m / z = 292.1 [M+H] + That is the case.

[0070] (2) Synthesis of I-2 [ka] (R)-4-(3-(1-methyl-1H-indazole-4-yl)phenyl)pyrrolidine-2-one [ka] To a solution of Pd(PPh3)4 (1.05 g, 0.91 mmol) in ethylene glycol dimethyl ether, a solution of (R)-4-(3-bromophenyl)pyrrolidine-2-one (4.51 g, 18.77 mmol) in ethylene glycol dimethyl ether (60 mL) was added and the mixture was stirred under a nitrogen atmosphere for 15 minutes. To the mixture from the previous step, a solution of 1-methyl-1H-indazole-4-boronic acid (5.00 g, 28.40 mmol) in ethanol (15 mL) was added and the mixture was stirred for 10 minutes. To the resulting mixture, a 2 M aqueous solution of Na2CO3 (80 mL) was added, and the mixture was dried under reflux for 20 hours. After the mixture cooled, the organic layer was separated, washed with brine, dried over sodium sulfate, and then concentrated under reduced pressure.

[0071] The residue was purified by silica gel column chromatography (hexane:benzene = 5:1) to obtain (R)-4-(3-(1-methyl-1H-indazole-4-yl)phenyl)pyrrolidine-2-one (5.25 g, 96%).

[0072] Liquid chromatography-mass spectrometry revealed m / z = 292.1 [M+H] + That is the case.

[0073] (3) Synthesis of I-3 [ka] (3-amino-1H-isoindole-7-yl)boronic acid [ka] Under a nitrogen atmosphere, 1.2 mL of anhydrous tetrahydrofuran solution of alkyllithium-tetramethylethylenediamine 1:1 complex (2.8 M hexane solution, 3.36 mmol) was added within 2.5 minutes to 20 mL of anhydrous tetrahydrofuran solution of 1H-isoindole-3-amine (445.5 mg, 3.37 mmol), and the mixture was stirred at -78°C for 30 minutes. 403.1 mg (3.88 mmol) of trimethyl borate was added to the mixture, and the mixture was stirred at -78°C for 5 minutes, after which the low-temperature constant-temperature water bath was stopped. After the mixture was allowed to return to room temperature, the tetrahydrofuran was removed under vacuum, a 5% aqueous HCl solution was added, and the mixture was washed with diethyl ether. The organic layer was separated, dried over magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate:hexane = 3:7) to obtain (3-amino-1H-isoindole-7-yl)boronic acid (474.5 mg, 80%).

[0074] Liquid chromatography-mass spectrometry revealed m / z = 177.1 [M+H] + That is the case.

[0075] (S)-4-(3-(3-amino-1H-isoindole-7-yl)phenyl)pyrrolidine-2-one [ka] To a solution of Pd(PPh3)4 (1.05 g, 0.91 mmol) in ethylene glycol dimethyl ether, a solution of (R)-4-(3-bromophenyl)pyrrolidine-2-one (4.51 g, 18.77 mmol) in ethylene glycol dimethyl ether (60 mL) was added and the mixture was stirred under a nitrogen atmosphere for 15 minutes. To the mixture from the previous step, a solution of (3-amino-1H-isoindole-7-yl)boronic acid (5.00 g, 28.40 mmol) in ethanol (15 mL) was added and the mixture was stirred for 10 minutes. To the resulting mixture, a 2 M aqueous solution of Na2CO3 (80 mL) was added, and the mixture was dried under reflux for 20 hours. After the mixture cooled, the organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:benzene = 5:1) to obtain (R)-4-(3-(3-amino-1H-isoindole-7-yl)phenyl)pyrrolidine-2-one (5.25 g, 96%).

[0076] Liquid chromatography-mass spectrometry revealed m / z = 292.1 [M+H] + That is the case.

[0077] (4) Synthesis of I-4 [ka] 3′-Bromo-[1,1′-biphenyl]-3-amine [ka] To a solution of Pd(PPh3)4 (1.05 g, 0.91 mmol) in ethylene glycol dimethyl ether, a solution of 3-iodoaniline (4.10 g, 18.77 mmol) in ethylene glycol dimethyl ether was added and the mixture was stirred under a nitrogen atmosphere for 15 minutes. To the mixture from the previous step, a solution of 3-bromophenylboronic acid (5.70 g, 28.40 mmol) in ethanol (15 mL) was added and the mixture was stirred for 10 minutes. To the resulting mixture, a 2 M aqueous solution of Na2CO3 (80 mL) was added, and the mixture was refluxed and dried for 20 hours. After cooling the mixture, the organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:benzene = 5:1) to obtain 3′-bromo-[1,1′-biphenyl]-3-amine (3.12 g, 67%).

[0078] Liquid chromatography-mass spectrometry revealed m / z = 248.0 [M+H] + That is the case.

[0079] 3′-(1-methyl-1H-indazole-4-yl)-[1,1′-biphenyl]-3-amine [ka] To a solution of Pd(PPh3)4 (1.05 g, 0.91 mmol) in ethylene glycol dimethyl ether, a solution of 3′-bromo-[1,1′-biphenyl]-3-amine (4.66 g, 18.77 mmol) in ethylene glycol dimethyl ether (60 mL) was added and the mixture was stirred under a nitrogen atmosphere for 15 minutes. To the mixture from the previous step, a solution of (1-methyl-1H-indazole-4-yl)boronic acid (5.00 g, 28.40 mmol) in ethanol (15 mL) was added and the mixture was stirred for 10 minutes. To the resulting mixture, a 2 M aqueous solution of Na2CO3 (80 mL) was added, and the mixture was dried under reflux for 20 hours. After the mixture cooled, the organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:benzene = 5:1) to obtain 3′-(1-methyl-1H-indazole-4-yl)-[1,1′-biphenyl]-3-amine (5.39 g, 96%).

[0080] Liquid chromatography-mass spectrometry revealed that the m / z ratio is 300.1[M+H]+.

[0081] (5) Synthesis of I-5 [ka] 3-(1-methyl-1H-indazole-4-yl)aniline [ka] To a solution of Pd(PPh3)4 (1.05 g, 0.91 mmol) in ethylene glycol dimethyl ether, a solution of 4-bromo-1-methyl-1H-indazole (3.96 g, 18.77 mmol) in ethylene glycol dimethyl ether (60 mL) was added and the mixture was stirred under a nitrogen atmosphere for 15 minutes. To the mixture from the previous step, a solution of 3-aminophenylboronic acid (3.89 g, 28.40 mmol) in ethanol (15 mL) was added and the mixture was stirred for 10 minutes. To the resulting mixture, a 2 M aqueous solution of Na2CO3 (80 mL) was added, and the mixture was refluxed and dried for 20 hours. After the mixture cooled, the organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:benzene = 5:1) to obtain 3-(1-methyl-1H-indazole-4-yl)aniline (4.02 g, 96%).

[0082] Liquid chromatography-mass spectrometry revealed m / z = 224.1 [M+H] + That is the case.

[0083] (S)-2-methyl-1-((3-(1-methyl-1H-indazole-4-yl)phenyl)amino)-2-butanol [ka] (R)-2-ethyl-2-methylethylene oxide (86.1 mg, 1.0 mmol), 3-(1-methyl-1H-indazole-4-yl)aniline (335.0 mg, 1.5 mmol), and reagent-grade DMF (6.7 mL) were added to a 20 mL pressure tube. The mixture was sealed at 60°C and stirred for 12 hours. After the reaction was cooled to room temperature, 6.7 mL of deionized water was added, the mixture was sealed at 60°C, and stirred for 12 hours. The solvent was removed using a rotary evaporator (22.5 mbar, 35°C), and the residue was purified by silica gel column chromatography (ethyl acetate:hexane = 1:2) to obtain (S)-2-methyl-1-((3-(1-methyl-1H-indazole-4-yl)phenyl)amino)-2-butanol (281.6 mg, 91%).

[0084] Liquid chromatography-mass spectrometry revealed m / z = 310.2 [M+H] + That is the case.

[0085] (6) Synthesis of I-6 [ka] N-(3-(1-methyl-1H-indazole-4-yl)phenyl)isobutylamide [ka] To a solution of Pd(PPh3)4 (1.05 g, 0.91 mmol) in ethylene glycol dimethyl ether, a solution of 4-bromo-1-methyl-1H-indazole (3.96 g, 18.77 mmol) in ethylene glycol dimethyl ether (60 mL) was added and the mixture was stirred under a nitrogen atmosphere for 15 minutes. To the mixture from the previous step, a solution of 3-isobutylamide phenylboronic acid (5.88 g, 28.40 mmol) in ethanol (15 mL) was added and the mixture was stirred for 10 minutes. To the resulting mixture, a 2 M aqueous solution of Na2CO3 (80 mL) was added, and the mixture was dried under reflux for 20 hours. After cooling the mixture, the organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:benzene = 5:1) to obtain N-(3-(1-methyl-1H-indazole-4-yl)phenyl)isobutylamide (5.29 g, 96%).

[0086] Liquid chromatography-mass spectrometry revealed m / z = 294.2 [M+H] + That is the case.

[0087] (7) Synthesis of I-7 [ka] (R)-2-methyl-4-(3-(1-methyl-1H-indazole-4-yl)phenyl)morpholine [ka] La[N(SiMe3)2]3 (124 mg, 0.20 mmol, stored and weighed in a fume hood) was mixed with CH2Cl2 (1.25 mL), 4-(3-fluorophenyl)-1-methyl-1H-indazole (200 μL, 0.20 mmol, in a 1.0 M CH2Cl2 solution), and (R)-2-methylmorpholine (37 μL, 0.22 mmol). The mixture was stirred at room temperature for 1 minute, and the residue was purified by silica gel column chromatography (CH2Cl2:MeOH = 95:5) to obtain (R)-2-methyl-4-(3-(1-methyl-1H-indazole-4-yl)phenyl)morpholine (57.2 mg, 93%).

[0088] Liquid chromatography-mass spectrometry revealed m / z = 308.2 [M+H] + That is the case.

[0089] (8) Synthesis of I-8 [ka] 2-(3-bromophenyl)oxazole-5-carboxylate ethyl ester [ka] To a solution of Pd(PPh3)4 (1.05 g, 0.91 mmol) in ethylene glycol dimethyl ether, a solution of 2-chlorooxazole-5-carboxylate ethyl ester (3.30 g, 18.77 mmol) (60 mL) was added and the mixture was stirred under a nitrogen atmosphere for 15 minutes. To the mixture from the previous step, a solution of 3-bromophenylboronic acid (5.70 g, 28.40 mmol) in ethanol (15 mL) was added and the mixture was stirred for 10 minutes. To the resulting mixture, a 2 M Na2CO3 aqueous solution (80 mL) was added, and the mixture was refluxed and dried for 20 hours. After cooling the mixture, the organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:benzene = 5:1) to obtain 2-(3-bromophenyl)oxazole-5-carboxylate ethyl ester (2.72 g, 49%).

[0090] Liquid chromatography-mass spectrometry revealed m / z = 296.0 [M+H] + That is the case.

[0091] (2-(3-bromophenyl)oxazol-5-yl)methanol [ka] To a methanol solution (100 mL, 0°C) of ethyl 2-(3-bromophenyl)oxazole-5-carboxylate (11.4 g, 38.6 mmol), NaBH4 (4.4 g, 115.8 mmol, 3 eq) was added in several portions. The mixture was stirred at room temperature for 3 hours, and after adding water (100 mL) to stop the reaction, the product was extracted with ethyl acetate (3 × 40 mL). The organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure to obtain (2-(3-bromophenyl)oxazole-5-yl)methanol (9.3 g, 95%).

[0092] Liquid chromatography-mass spectrometry revealed m / z = 254.0 [M+H] + That is the case.

[0093] (2-(3-(1-methyl-1H-indazole-4-yl)phenyl)oxazol-5-yl)methanol [ka] To a solution of Pd(PPh3)4 (1.05 g, 0.91 mmol) in ethylene glycol dimethyl ether, a solution of (2-(3-bromophenyl)oxazole-5-yl)methanol (4.77 g, 18.77 mmol) in ethylene glycol dimethyl ether (60 mL) was added and the mixture was stirred under a nitrogen atmosphere for 15 minutes. To the mixture from the previous step, a solution of (1-methyl-1H-indazole-4-yl)boronic acid (5.00 g, 28.40 mmol) in ethanol (15 mL) was added and the mixture was stirred for 10 minutes. To the resulting mixture, a 2 M aqueous solution of Na2CO3 (80 mL) was added, and the mixture was dried under reflux for 20 hours. After the mixture cooled, the organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:benzene = 5:1) to obtain (2-(3-(1-methyl-1H-indazole-4-yl)phenyl)oxazol-5-yl)methanol (5.50 g, 96%).

[0094] Liquid chromatography-mass spectrometry revealed m / z = 306.1 [M+H] + That is the case.

[0095] (9) Synthesis of I-9 [ka] (3-(1-methyl-1H-indazole-4-yl)phenyl)methanol [ka] To a solution of Pd(PPh3)4 (1.05 g, 0.91 mmol) in ethylene glycol dimethyl ether, a solution of 4-bromo-1-methyl-1H-indazole (3.96 g, 18.77 mmol) in ethylene glycol dimethyl ether (60 mL) was added and the mixture was stirred under a nitrogen atmosphere for 15 minutes. To the mixture from the previous step, a solution of 3-hydroxymethylphenylboronic acid (4.32 g, 28.40 mmol) in ethanol (15 mL) was added and the mixture was stirred for 10 minutes. To the resulting mixture, a 2 M aqueous solution of Na2CO3 (80 mL) was added, and the mixture was refluxed and dried for 20 hours. After cooling the mixture, the organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:benzene = 5:1) to obtain (3-(1-methyl-1H-indazole-4-yl)phenyl)methanol (4.29 g, 96%).

[0096] Liquid chromatography-mass spectrometry revealed m / z = 239.1 [M+H] + That is the case.

[0097] 2-Methyl-1-((3-(1-methyl-1H-indazole-4-yl)phenyl)oxy)isopropanol [ka] 2,2-Epoxybutane (15.0 mL, 168 mmol) and (3-(1-methyl-1H-indazole-4-yl)phenyl)methanol (11.0 g, 46 mmol) were dissolved in toluene (40 mL), to which 50% aq NaOH (12 mL) was added and the mixture was stirred at 100 °C for 30 hours. Water and ethyl acetate were added to the reaction mixture, the organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to obtain 2-methyl-1-((3-(1-methyl-1H-indazole-4-yl)phenyl)oxy)isopropanol (14.2 g, 99%).

[0098] Liquid chromatography-mass spectrometry revealed m / z = 311.2 [M+H]+ That is the case.

[0099] (10) Synthesis of I-10 [ka] 4-(3-((1H-pyrazole-4-yl)oxy)phenyl)-1-methyl-1H-indazole [ka] 4-hydroxypyrazole (849.4 mg, 10.1 mmol) was mixed with sodium hydride (60%, 256.8 mg, 10.7 mmol). After 15 minutes, 4-(3-fluorophenyl)-1-methyl-1H-indazole (2.3 g, 10.2 mmol) was added, and the mixture was stirred at room temperature for 2 hours. The product was extracted with ethyl acetate, and the organic layer was separated and dried. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 1:1) to obtain 4-(3-((1H-pyrazole-4-yl)oxy)phenyl)-1-methyl-1H-indazole (1.47 g, 50%).

[0100] Liquid chromatography-mass spectrometry revealed m / z = 291.1 [M+H] + That is the case.

[0101] (11) Synthesis of I-11 [ka] (5-(3-(1-methyl-1H-indazole-4-yl)phenyl)oxazole-4-yl)methanol [ka] To a solution of Pd(PPh3)4 (1.05 g, 0.91 mmol) in ethylene glycol dimethyl ether, a solution of (5-(3-chlorophenyl)oxazole-4-yl)methanol (3.93 g, 18.77 mmol) in ethylene glycol dimethyl ether (60 mL) was added and the mixture was stirred under a nitrogen atmosphere for 15 minutes. To the mixture from the previous step, a solution of (1-methyl-1H-indazole-4-yl)boronic acid (5.00 g, 28.40 mmol) in ethanol (15 mL) was added and the mixture was stirred for 10 minutes. To the resulting mixture, a 2 M aqueous solution of Na2CO3 (80 mL) was added, and the mixture was dried under reflux for 20 hours. After the mixture cooled, the organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:benzene = 5:1) to obtain (5-(3-(1-methyl-1H-indazole-4-yl)phenyl)oxazol-4-yl)methanol (5.50 g, 96%).

[0102] Liquid chromatography-mass spectrometry revealed m / z = 306.1 [M+H] + That is the case.

[0103] (12) Synthesis of I-12 [ka] 2-(3-chlorophenyl)-4-hydroxypyridine [ka] While stirring, Aliquat-336 (10% of substrate weight) was added in a single batch to a mixture of 2-(3-chlorophenyl)-4-methoxypyridine (20 mmol) and 47% HBr (4.5 mmol). The mixture was heated to 105±5°C, and the reaction was continuously monitored by TLC. After the reaction was complete, the mixture was cooled to room temperature, and water (25 mL) was added to stop the reaction. The product was extracted with ethyl acetate (3 × 30 mL), the organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:ethyl acetate = 2:1) to obtain 2-(3-chlorophenyl)-4-hydroxypyridine (3.94 g, 96%).

[0104] Liquid chromatography-mass spectrometry revealed m / z = 206.0 [M+H] + That is the case.

[0105] 2-(3-(1-methyl-1H-indazole-4-yl)phenyl)-4-hydroxypyridine [ka]

[0106] To a solution of Pd(PPh3)4 (1.05 g, 0.91 mmol) in ethylene glycol dimethyl ether, a solution of 2-(3-chlorophenyl)-4-hydroxypyridine (3.86 g, 18.77 mmol) in ethylene glycol dimethyl ether (60 mL) was added and the mixture was stirred under a nitrogen atmosphere for 15 minutes. To the mixture from the previous step, a solution of (1-methyl-1H-indazole-4-yl)boronic acid (5.00 g, 28.40 mmol) in ethanol (15 mL) was added and the mixture was stirred for 10 minutes. To the resulting mixture, a 2 M aqueous solution of Na2CO3 (80 mL) was added, and the mixture was dried under reflux for 20 hours. After cooling the mixture, the organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:benzene = 5:1) to obtain 2-(3-(1-methyl-1H-indazole-4-yl)phenyl)-4-hydroxypyridine (5.43 g, 96%).

[0107] Liquid chromatography-mass spectrometry revealed m / z = 302.1 [M+H] + That is the case.

[0108] (13) Synthesis of I-13 [ka] (3-(1-methyl-1H-indazole-4-yl)phenyl)methanol [ka] To a solution of Pd(PPh3)4 (1.05 g, 0.91 mmol) in ethylene glycol dimethyl ether, a solution of 4-bromo-1-methyl-1H-indazole (3.96 g, 18.77 mmol) in ethylene glycol dimethyl ether (60 mL) was added and the mixture was stirred under a nitrogen atmosphere for 15 minutes. To the mixture from the previous step, a solution of 3-hydroxymethylphenylboronic acid (4.32 g, 28.40 mmol) in ethanol (15 mL) was added and the mixture was stirred for 10 minutes. To the resulting mixture, a 2 M aqueous solution of Na2CO3 (80 mL) was added, and the mixture was refluxed and dried for 20 hours. After cooling the mixture, the organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:benzene = 5:1) to obtain (3-(1-methyl-1H-indazole-4-yl)phenyl)methanol (4.29 g, 96%).

[0109] Liquid chromatography-mass spectrometry revealed m / z = 239.1 [M+H] + That is the case.

[0110] 1-Methyl-4-(3-((4-methyl-1H-pyrazole-1-yl)methyl)phenyl)-1H-indazole [ka] In a 100 mL round-bottom flask, 2.38 g (10 mmol) of (3-(1-methyl-1H-indazole-4-yl)phenyl)methanol, 1.64 g (20 mmol) of 4-methyl-1H-pyrazole, and 20.26 g (1 mmol) of Ni(ClO4) were added. Finally, 20 mL of anhydrous 1,2-dichloroethane was added, and the mixture was stirred at 85°C for 6 hours. After the mixture was cooled to room temperature, it was poured into an ice water mixture, and the product was extracted with dichloromethane (3 × 50 mL). The organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:benzene = 5:1) to obtain 1-methyl-4-(3-((4-methyl-1H-pyrazole-1-yl)methyl)phenyl)-1H-indazole (2.78 g, 92%).

[0111] Liquid chromatography-mass spectrometry revealed m / z = 303.1 [M+H] + That is the case.

[0112] (14) Synthesis of I-14 [ka] 2-Methyl-N-(3-(1-methyl-1H-indazole-4-yl)phenyl)isopropylamine [ka] To a solution of Pd(PPh3)4 (1.05 g, 0.91 mmol) in ethylene glycol dimethyl ether, a solution of N-(3-bromobenzyl)-2-methylisopropylamine (4.55 g, 18.77 mmol) in ethylene glycol dimethyl ether (60 mL) was added and the mixture was stirred under a nitrogen atmosphere for 15 minutes. To the mixture from the previous step, a solution of (1-methyl-1H-indazole-4-yl)boronic acid (5.00 g, 28.40 mmol) in ethanol (15 mL) was added and the mixture was stirred for 10 minutes. To the resulting mixture, a 2 M aqueous solution of Na2CO3 (80 mL) was added, and the mixture was dried under reflux for 20 hours. After the mixture cooled, the organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:benzene = 5:1) to obtain 2-methyl-N-(3-(1-methyl-1H-indazole-4-yl)phenyl)isopropylamine (5.29 g, 96%).

[0113] Liquid chromatography-mass spectrometry revealed m / z = 294.2 [M+H] + That is the case.

[0114] (15) Synthesis of I-15 [ka] 1-(chloromethyl)-3-iodobenzene [ka] Under a nitrogen atmosphere, (3-iodophenyl)methanol (41.2 g), toluene (300 mL), and pyridine (0.5 mL) were mixed and stirred at 45°C for 1 hour. Thionyl dichloride (14.0 mL) was added to the mixture at a temperature of 45-55°C and refluxed for 2 hours. The mixture was cooled to 25°C, water (300 mL) and toluene (300 mL) were added and mixed, the organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (n-heptane:toluene = 1:1) to obtain 1-(chloromethyl)-3-iodobenzene (42.79 g, 94%).

[0115] Liquid chromatography-mass spectrometry revealed m / z = 252.9 [M+H] + That is the case.

[0116] 3-(3-iodophenyl)-1-methylazetidine-3-ol [ka] To a 190 mL diethyl ether solution of magnesium powder (3.2 g, 131.66 mmol), a 32 mL diethyl ether solution of 1-(chloromethyl)-3-iodobenzene (31.97 g, 126.62 mmol) was added and the mixture was stirred at 45°C for 4 hours. After the reaction mixture was cooled to room temperature, a 32 mL diethyl ether solution of 1-methylazetidine-3-one (16.81 g, 197.49 mmol) at 0°C was added and the mixture was stirred overnight at room temperature. The mixture was placed on ice and dilute hydrochloric acid (60 mL) and NaHSO4 solution (1.0 M, 50 mL) were added. The product was extracted with ethyl acetate (3 × 100 mL), the organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 1:1) to obtain 3-(3-iodophenyl)-1-methylazetidine-3-ol (36.46 g, 95%).

[0117] Liquid chromatography-mass spectrometry revealed m / z = 304.0 [M+H] + That is the case.

[0118] 1-Methyl-3-(3-(1-methyl-1H-indazole-4-yl)phenyl)azetidine-3-ol [ka] To a solution of Pd(PPh3)4 (1.05 g, 0.91 mmol) in ethylene glycol dimethyl ether, a solution of 3-(3-iodophenyl)-1-methylazetidine-3-ol (5.69 g, 18.77 mmol) in ethylene glycol dimethyl ether (60 mL) was added and the mixture was stirred under a nitrogen atmosphere for 15 minutes. To the mixture from the previous step, a solution of (1-methyl-1H-indazole-4-yl)boronic acid (5.00 g, 28.40 mmol) in ethanol (15 mL) was added and the mixture was stirred for 10 minutes. To the resulting mixture, a 2 M aqueous solution of Na2CO3 (80 mL) was added, and the mixture was refluxed and dried for 20 hours. After the mixture cooled, the organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:benzene = 5:1) to obtain 1-methyl-3-(3-(1-methyl-1H-indazole-4-yl)phenyl)azetidine-3-ol (5.54 g, 96%).

[0119] Liquid chromatography-mass spectrometry revealed m / z = 308.2 [M+H] + That is the case.

[0120] (16) Synthesis of I-16 [ka] 3-(3-chlorophenyl)-N-ethylpropionamide [ka] Under a nitrogen atmosphere, a solution of methyllithium in diethyl ether (1.6 M, 0.8 mL, 0.5 mmol) was added to a mixture of copper iodide and anhydrous tetrahydrofuran (12 mL) and mixed, then stirred in an ice bath for 15 minutes. The ice bath was changed to -78°C, and anhydrous hexamethyltriamine phosphate (1.7 mL, 10 mmol) was added with stirring, followed by a solution of diisobutylaluminum hydride in cyclohexane (1 M, 8.0 mL, 8 mmol), and the mixture was stirred for 70 minutes while maintaining the temperature. (E)-3-(3-chlorophenyl)-N-ethylacrylamide (1.0 mmol) was added, and the mixture was stirred for 70 minutes while maintaining the temperature. Methanol (5 mL) was added, and after the mixture had returned to room temperature naturally, saturated potassium sodium citrate (20 mL) and diethyl ether (30 mL) were added, and the mixture was stirred overnight at room temperature. The organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 1:1) to obtain 3-(3-chlorophenyl)-N-ethylpropionamide (203.2 mg, 96%).

[0121] Liquid chromatography-mass spectrometry revealed m / z = 212.1 [M+H] + That is the case.

[0122] N-ethyl-3-(3-(1-methyl-1H-indazole-4-yl)phenyl)propionamide [ka] To a solution of Pd(PPh3)4 (1.05 g, 0.91 mmol) in ethylene glycol dimethyl ether, a solution of 3-(3-chlorophenyl)-N-ethylpropionamide (3.97 g, 18.77 mmol) in ethylene glycol dimethyl ether (60 mL) was added and the mixture was stirred under a nitrogen atmosphere for 15 minutes. To the mixture from the previous step, a solution of (1-methyl-1H-indazole-4-yl)boronic acid (5.00 g, 28.40 mmol) in ethanol (15 mL) was added and the mixture was stirred for 10 minutes. To the resulting mixture, a 2 M aqueous solution of Na2CO3 (80 mL) was added, and the mixture was dried under reflux for 20 hours. After the mixture cooled, the organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:benzene = 5:1) to obtain N-ethyl-3-(3-(1-methyl-1H-indazole-4-yl)phenyl)propionamide (5.54 g, 96%).

[0123] Liquid chromatography-mass spectrometry revealed m / z = 308.2 [M+H] + That is the case.

[0124] (17) Synthesis of I-17 [ka] 4-(3-chlorophenyl)-1-methylindazole [ka] To a solution of Pd(PPh3)4 (1.05 g, 0.91 mmol) in ethylene glycol dimethyl ether, a solution of 4-bromo-1-methyl-1H-indazole (3.96 g, 18.77 mmol) in ethylene glycol dimethyl ether (60 mL) was added and the mixture was stirred under a nitrogen atmosphere for 15 minutes. To the mixture from the previous step, a solution of (3-chlorophenyl)boronic acid (4.44 g, 28.40 mmol) in ethanol (15 mL) was added and the mixture was stirred for 10 minutes. To the resulting mixture, a 2 M aqueous solution of Na2CO3 (80 mL) was added, and the mixture was refluxed and dried for 20 hours. After cooling the mixture, the organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:benzene = 5:1) to obtain 4-(3-chlorophenyl)-1-methylindazole (4.37 g, 96%).

[0125] Liquid chromatography-mass spectrometry revealed m / z = 243.1 [M+H] + That is the case.

[0126] 4-(3-(1-methylindazole-4-yl)phenyl)pyridine-3-amine [ka] To a solution of Pd(PPh3)4 (1.05 g, 0.91 mmol) in ethylene glycol dimethyl ether, a solution of 4-(3-chlorophenyl)-1-methylindazole (4.37 g, 18.02 mmol) in ethylene glycol dimethyl ether (60 mL) was added and the mixture was stirred under a nitrogen atmosphere for 15 minutes. To the mixture from the previous step, a solution of (3-aminopyridine-4-yl)boronic acid (3.92 g, 28.40 mmol) in ethanol (15 mL) was added and the mixture was stirred for 10 minutes. To the resulting mixture, a 2 M aqueous solution of Na2CO3 (80 mL) was added, and the mixture was refluxed and dried for 20 hours. After the mixture cooled, the organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:benzene = 5:1) to obtain 4-(3-(1-methylindazole-4-yl)phenyl)pyridine-3-amine (5.19 g, 96%).

[0127] Liquid chromatography-mass spectrometry revealed m / z = 301.1 [M+H] + That is the case.

[0128] (18) Synthesis of I-18 [ka] 3-(3-chlorophenyl)pyrazine-2-ol [ka] While stirring, Aliquat-336 (10% of substrate weight) was added in a single batch to a mixture of 2-(3-chlorophenyl)-3-methoxypyrazine (20 mmol) and 47% HBr (4.5 mmol). The mixture was heated to 105±5°C, and the reaction was continuously monitored by TLC. After the reaction was complete, the mixture was cooled to room temperature, and water (25 mL) was added to stop the reaction. The product was extracted with ethyl acetate (3 × 30 mL), the organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:ethyl acetate = 2:1) to obtain 3-(3-chlorophenyl)pyrazine-2-ol (3.96 g, 96%).

[0129] Liquid chromatography-mass spectrometry revealed m / z = 207.0 [M+H] + That is the case.

[0130] 3-(3-(1-methylindazole-4-yl)phenyl)pyrazine-2-phenol [ka] To a solution of Pd(PPh3)4 (1.05 g, 0.91 mmol) in ethylene glycol dimethyl ether, a solution of 3-(3-chlorophenyl)pyrazine-2-ol (3.87 g, 18.77 mmol) in ethylene glycol dimethyl ether (60 mL) was added and the mixture was stirred under a nitrogen atmosphere for 15 minutes. To the mixture from the previous step, a solution of (1-methyl-1H-indazole-4-yl)boronic acid (5.00 g, 28.40 mmol) in ethanol (15 mL) was added and the mixture was stirred for 10 minutes. To the resulting mixture, a 2 M aqueous solution of Na2CO3 (80 mL) was added, and the mixture was refluxed and dried for 20 hours. After cooling the mixture, the organic layer was separated, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane:benzene = 5:1) to obtain 3-(3-(1-methylindazole-4-yl)phenyl)pyrazine-2-phenol (5.44 g, 96%). Liquid chromatography-mass spectrometry revealed m / z = 303.1 [M+H] + That is the case.

[0131] (Example 1) Virtual screening of target kinase and biphenyl derivative molecules Virtual screening is a computational technique used in the drug discovery process to search a library of small molecule compounds using computational methods and obtain chemical small molecules that can bind to drug targets. Compared to conventional high-throughput screening techniques, virtual screening has the advantages of being highly efficient and economical. Therefore, in this application, the action of compounds was predicted using virtual screening. The specific theory and methods were referred to in "Docking Screens for Novel Ligands Conferring New Biology" (Irwin JJ and Shoichet BK, J Med Chem. 2016 May 12;59(9):4103-20).

[0132] Comparative protein structure analysis revealed that PDGFR, TNIK, CLK4, and CLK3 have similar three-dimensional structures (RMSD ≤ 8.4), with the structural similarity between PDGFR and CLK3 (RMSD = 8.4) being lower than that between other proteins (RMSD ≤ 2.9) (Figure 1A). In the protein structures, the positions of important sites in the active sites of each kinase were highly conserved (Figure 1B). Based on these analysis results, it is considered that there is a structural basis for the simultaneous inhibition of PDGFR, TNIK, and CLK by the biphenyl derivative of this application.

[0133] Using AutoDock Vina software, biphenyl derivative molecules I-1 to I-18 were molecularly docked with multiple target proteins, and the binding energy and ligand efficiency were calculated for each biphenyl derivative molecule and target protein docking result. The specific docking results are shown in Table 1. Columns 2 to 5 of Table 1 show the binding free energy (BFS) of the compounds in this application with different target proteins, calculated by the docking software. A larger absolute value indicates stronger binding ability between the small molecule ligand and the target protein. When the binding energy levels of the compounds targeted in this application were predicted, all of the compounds in this application significantly exceeded the threshold of 3 set based on the target characteristics. This result indicates that all of the biphenyl derivative molecules in this application have the ability to bind to their targets.

[0134] Table 1. Energy analysis of the binding process between the compound of this application and the target protein. [Table 1]

[0135] Furthermore, molecular dynamics simulations were performed on the biphenyl derivative molecules in this application. Molecular dynamics simulation is a computational method that simulates and predicts the behavior of molecular systems by numerical integration based on the principles of Newtonian mechanics and intermolecular interaction force fields. Molecular dynamics simulations provide detailed information on the structure, dynamics, and function of proteins, and contribute to elucidating protein conformational changes, binding site characteristics, and interactions with small molecules. By simulating intermolecular interactions, it is possible to screen potential drug molecules, predict the mode of interaction with proteins, and evaluate binding ability. Therefore, in this application, the chemical basis for the binding of biphenyl derivatives was verified by molecular dynamics simulation. The principle and method were referred to in "Molecular Dynamics Simulations of Protein-Drug Complexes: A Computational Protocol for Investigating the Interactions of Small-Molecule Therapeutics with Biological Targets and Biosensors" (Hadden JA and Perilla JR, Methods Mol Biol. 2018;1762:245-270).

[0136] The stability of each complex system was investigated using Amber molecular dynamics simulation software. In the molecular dynamics simulation experiments, ff14SB and Gaff force fields were applied to the receptor protein and ligand small molecule, respectively. Input files were created using the Antechamber and Leap programs of the Amber tool, and then a 100 ns simulation calculation was performed. To simulate physiological salt concentration and neutralize the entire system, 0.15 M Na+ and Cl- ions were introduced into the simulation system, and a 15 Å side TIP3P water box was used. Bond lengths including all hydrogen bonds were constrained using the SHAKE algorithm, and the particle mesh Ewald method was used to calculate long-range electrostatic interactions. The system underwent 10,000 steps of minimization followed by 10,000 steps of equilibration. The system temperature was gradually equilibrated from 200 K to 300 K via 5,000 steps. After equilibration, the prepared complex system was subjected to molecular dynamics simulation at a constant temperature of 300 K and under 1 atmosphere.

[0137] Molecular dynamics simulations showed that during a 100 ns simulation, the complex structures of each small molecule and protein target gradually reached a stable state (Figures 2-10, Series A). Table 1 shows the binding energies of each small molecule and protein target calculated using the MMGPSA program of the Amber tool. Analysis of the energy contribution of amino acid sites in each protein target to ligand small molecule binding showed that I-1, I-4, and I-12 have similar binding sites in the same target (Figures 2-10, Series B). On the other hand, even with the same amino acid site, differences were observed in the energy contribution to binding of different small molecules. For example, the energy contributions of I-1, I-4, and I-12 to the binding of LYS43 in TNIK were -0.39 kJ / mol, -2.36 kJ / mol, and -0.58 kJ / mol, respectively, which may be one of the factors contributing to the differences in TNIK's sensitivity to these three molecules. Analysis of the stabilized complex structure using molecular dynamics simulations revealed that each small molecule is stably bound to the active catalytic center of the kinase target, and that the amino acid sites involved in ligand binding (with a binding energy contribution of less than -0.5 kJ / mol) interact directly with the small molecules primarily through hydrogen bonding and hydrophobicity (Figures 2-10, C series).

[0138] As shown in Figures 2 to 4, Series A shows the changes in RMSD of the TNIK-I-1, I-4, and I-12 complexes over simulation time during the simulation process, indicating that TNIK can bind to any of I-1, I-4, and I-12, and that the formed complex structure is in a stable binding state. Series B shows the energy decomposition diagrams of the interaction between each amino acid site of TNIK and I-1, I-4, and I-12, with the x-axis representing the amino acid sequence of TNIK and the y-axis representing the energy contribution of each amino acid site to ligand binding. The names of amino acid sites with a ligand binding energy contribution of less than -0.5 kJ / mol are displayed. These results indicate that the binding sites of TNIK-I-1, I-4, and I-12 are similar. Furthermore, Series C shows the stabilized complex structures of TNIK-I-1, I-4, and I-12 as determined by molecular dynamics simulations. Proteins are shown as cartoon models, with the color of each site indicating the energy contribution to ligand binding. Small molecules and the main chains or side chains of the residues interacting with them are shown as stick models, with hydrogen bonding interactions indicated by long dashed lines and hydrophobic interactions by short dashed lines. These results clearly demonstrate the chemical basis of the active catalytic centers of the kinase targets to which TNIK binds with I-1, I-4, and I-12, and their interactions, indicating a consistent basis for chemical bond formation.

[0139] As shown in Figures 5 to 7, Series A shows the changes in RMSD of the PDGFRα and I-1, I-4, and I-12 complexes over simulation time during the simulation process, indicating that PDGFRα can bind to any of I-1, I-4, and I-12, and that the formed complex structure is in a stable binding state. Series B shows the energy decomposition diagrams of the interaction between each amino acid site of PDGFRα and I-1, I-4, and I-12, with the x-axis representing the amino acid sequence of PDGFRα and the y-axis representing the energy contribution of each amino acid site to ligand binding. The names of amino acid sites with an energy contribution of less than -0.5 kJ / mol to ligand binding are displayed. These results indicate that the binding sites of PDGFRα and I-1, I-4, and I-12 are similar. Furthermore, Series C shows the complex structures of PDGFRα and I-1, I-4, and I-12 stabilized by molecular dynamics simulations. Proteins are shown as cartoon models, with the color of each site indicating the energy contribution to ligand binding. Small molecules and the main chains or side chains of residues interacting with them are shown as stick models, with hydrogen bonding interactions indicated by long dashed lines and hydrophobic interactions by short dashed lines. These results clearly demonstrate the chemical basis of the active catalytic centers of kinase targets to which PDGFRα binds with I-1, I-4, and I-12, and their interactions, indicating a consistent basis for chemical bond formation.

[0140] As shown in Figures 8 to 10, Series A shows the changes in RMSD of the CLK4 and I-1, I-4, and I-12 complexes over simulation time during the simulation process, indicating that CLK4 can bind to any of I-1, I-4, and I-12, and that the formed complex structure is in a stable binding state. Series B shows the energy decomposition diagrams of the interaction between each amino acid site of CLK4 and I-1, I-4, and I-12, with the x-axis representing the amino acid sequence of CLK4 and the y-axis representing the energy contribution of each amino acid site to ligand binding. The names of amino acid sites with an energy contribution of less than -0.5 kJ / mol to ligand binding are displayed. These results indicate that the binding sites of CLK4 and I-1, I-4, and I-12 are similar. Furthermore, Series C shows the complex structures of CLK4 and I-1, I-4, and I-12 stabilized by molecular dynamics simulations. Proteins are shown as cartoon models, with the color of each site indicating the energy contribution to ligand binding. Small molecules and the main chains or side chains of residues interacting with them are shown as stick models, with hydrogen bonding interactions indicated by long dashed lines and hydrophobic interactions by short dashed lines. These results clearly demonstrate the chemical basis of the active catalytic centers of kinase targets to which CLK4 binds with I-1, I-4, and I-12, and their interactions, indicating a consistent basis for chemical bond formation.

[0141] As shown in Figures 11 and 12, Series A shows the changes in RMSD of the JNK1 and I-1 and I-4 complexes over simulation time during the simulation process, indicating that JNK1 can bind to either I-1 or I-4, and that the formed complex structure is in a stable binding state. Series B shows the energy decomposition diagrams of the interaction between each amino acid site of JNK1 and I-1 and I-4, with the x-axis representing the amino acid sequence of JNK1 and the y-axis representing the energy contribution of each amino acid site to ligand binding. The names of amino acid sites with an energy contribution of less than -0.5 kJ / mol to ligand binding are displayed. These results indicate that the binding sites of JNK1 and I-1 and I-4 are similar. Furthermore, Series C shows the complex structures formed by JNK1, I-1, and I-4 stabilized by molecular dynamics simulations. Proteins are shown as cartoon models, with the color of each site indicating the energy contribution to ligand binding. Small molecules and the main chains or side chains of residues interacting with them are shown as stick models, with hydrogen bonding interactions indicated by long dashed lines and hydrophobic interactions by short dashed lines. These results clearly demonstrate the chemical basis of the active catalytic center of the kinase target to which JNK1 and I-1 and I-4 bind, and their interactions, indicating a consistent basis for chemical bond formation.

[0142] Based on the above, it has been shown that the biphenyl derivatives of this application have a structure capable of binding to a series of kinase targets and possess a chemical basis for stable binding.

[0143] (Example 2) Biochemical screening of biphenyl derivatives and target kinases Based on the results of Example 1, in vitro kinase profile screening was performed on the biphenyl derivatives of this application to further confirm the actual function of the biphenyl derivatives and target kinases. Screening for activity inhibition effects was performed on 330 kinases. In the kinase activity test, first, the kinase (15-50 nM, 2.5 μL, prepared in test buffer) and biphenyl derivative I-1 (10 mM, 25 nL, prepared in DMSO) were mixed and incubated at 25°C for 10 minutes. Subsequently, a mixture of kinase peptide substrate (0.2 mg / mL, supplier: GenScript) and ATP (10-60 μM, supplier: Promega, product number: V915B) (total 2.5 μL, prepared in test buffer) was added to the mixture and reacted at 25°C for 60-120 minutes. Finally, the reaction product was quantitatively analyzed using HTRF or ADP-Glo ​​method. The test buffer consisted of 50 mM HEPES (supplier: Thermo Fisher, product number: 15630080), 10 mM MgCl2 (supplier: Sigma, product number: M1028), 0.01% Brij35 (supplier: Millipore, product number: 1018940100), 1 mM EGTA (supplier: Sigma, product number: E3889), and 2 mM DTT (supplier: MCE, product number: HY-15917). During the kinase activity tests, the consumption of each substrate was less than 10%. Each kinase activity test was performed with two technical replicates. As a result, biphenyl derivative I-1 (Figures 13-14) showed significant inhibitory activity against multiple target kinases. This result further supports the calculations and predictions in Example 1.

[0144] (Example 3) IC between a biphenyl derivative and a target kinase 50 screening To further evaluate the binding ability of biphenyl derivatives to target kinases, IC 50 The inhibitory effect of biphenyl derivatives on target kinase activity was evaluated by measurement. IC in this experiment 50IC refers to the concentration of a drug or inhibitor required to suppress the activity of a given kinase enzyme by half, and in pharmacy, it is an indicator of the antagonistic ability of an antagonist in in vitro experiments. 50 The specific operating procedures were carried out by referring to “Assessing the Inhibitory Potential of Kinase Inhibitors In Vitro: Major Pitfalls and Suggestions for Improving Comparability of Data Using CK1 Inhibitors as an Example” (Roth A et al., Molecules. 2021 Aug 12;26(16):4898).

[0145] First, a mixture of 2×ATP and substrate, and a mixture of 2×TNIK and MgCl2 were prepared using test buffer. Next, 40 nL of small molecules and 2 μL of the 2×TNIK and MgCl2 mixture were transferred to a 384-well plate and incubated at room temperature for 10 minutes after mixing. Subsequently, 2 μL of the 2×ATP and substrate mixture was transferred to the 384-well plate and reacted at room temperature for 60 minutes. 4 μL of ADP-Glo ​​reagent (supplier: Promega, product number: V9103) was added to the reaction system and incubated at room temperature for 40 minutes. Finally, 8 μL of kinase detection reagent (supplier: Promega, product number: V9103) was added to the reaction system and incubated at room temperature for 40 minutes, after which the fluorescence signal was read using a microplate reader. The concentration gradients for biphenyl derivatives I-1, I-4, and I-12 were set to 50,000 μM, 12,500 μM, 3,125 μM, 781.3 μM, 195.3 μM, 48.83 μM, 12.21 μM, 3.05 μM, 0.76 μM, and 0.19 μM.

[0146] The inhibition rate (%) was calculated using the formula: Inhibition rate % = 100% - (Small molecule reading - Positive control reading) / (Negative control reading - Positive control reading) * 100%. The logarithm values ​​of the inhibition rate (%) and inhibitor concentration were substituted into a nonlinear regression equation to obtain the IC of the small molecule. 50 :Y=Bottom+(Top-Bottom) / (1+10^((LogIC50 The formula -X*hillslope) was calculated, where X is the logarithm of the inhibitor concentration and Y is the inhibition rate (%). As shown in Figure 15, the biphenyl derivatives I-1, I-4, and I-12 all showed binding ability to the target kinase and exhibited significant antagonistic activity against the target kinase.

[0147] Combining the results of Examples 2 and 3, the above results indicate that both biphenyl derivatives can bind to the target kinase.

[0148] (Example 4) Biphenyl derivatives can induce the simultaneous expression of Oct4, Lin28A, and c-Myc reprogramming core genes. CHIR99021 is a widely used WNT pathway regulatory compound and has a very significant effect on maintaining pluripotency in mammalian embryos (Meek et al., STEMCELLS. 2007, 31, 10, pp. 2104-2115), and is therefore considered a potential reprogramming inducer. To further investigate whether biphenyl derivatives can initiate cell reprogramming, CHIR99021 was used as a control in this application.

[0149] Human mesenchymal cells were cultured in T25 flasks, seeded at 4 × 10⁵ cells, and cultured in serum-free Dulbecco's modified Eagle medium (DMEM-F12 medium) with 20 μM of the biphenyl derivative added to each medium. The culture conditions were 37°C and 5% carbon dioxide. On day 3, total RNA was extracted using the RNeasy Mini or Micro Kit (QIAGEN), and cDNA was synthesized from 1 mg of RNA using the SuperScript III First-Strand Synthesis System (Invitrogen). Quantitative PCR labeling and reaction were performed using SYBR Premix Ex Taq (TaKaRa) and Thermal Cycler Dice Real Time System (TaKaRa), with beta-Actin used as the internal standard. All data were analyzed using the delta-Ct method. Repeated studies were performed in three groups for each test, and analysis of variance was performed. Primer sequences for identifying genes encoding different cell markers are shown in Table 2. As shown in Figure 16, compared to a blank control group (CK) that did not use biphenyl derivative small molecules and a control group to which CHIR99021 was added alone, only the biphenyl derivative was able to simultaneously induce and express Oct4, Lin28A, and c-Myc. Therefore, the use of biphenyl derivatives alone can significantly increase the expression of multiple reprogramming core genes. Furthermore, the simultaneous expression of the reprogramming core gene group, Oct4, Lin28A, and c-Myc, is a necessary condition for various somatic cell reprogramming processes.

[0150] Table 2. Primer sequences for QPCR of compound effector genes. [Table 2]

[0151] The technical features of the above-described embodiments can be arbitrarily combined. For the sake of brevity of description, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should all be regarded as being within the scope described in this specification.

[0152] The above-described embodiments merely represent some embodiments of this application. Although the description is relatively specific and detailed, it should by no means be understood as limiting the scope of the patent for the invention. It should be noted that those skilled in the art can make some modifications and improvements without departing from the concept of this application, and all of them belong to the protection scope of this application. Therefore, the scope of protection sought by the patent of this application should be determined based on the appended claims, and the specification and drawings are used to interpret the content of the claims.

[0153] (Appendix 1) A biphenyl derivative having a structure represented by Formula I, or a pharmaceutically acceptable salt, ester, amide, solvate, active metabolite, polymorph, isotope-labeled, isomer, or prodrug of the structure represented by Formula I. [Chemical formula] (In the formula, Ring A is a five-membered ring substituted or unsubstituted with a methyl group or an amino group, and one or two nitrogen atoms are included in the ring atoms of Ring A, and R 1 is a hydrogen bond donor or acceptor, and contains one or more of an amino group, an imino group, a hydroxy group, and an ether bond in its structure.)

[0154] (Appendix 2) The above R 1 is selected from any one of the following structures, and the biphenyl derivative according to Appendix 1 is characterized in that. [Chemical formula] ("*" indicates the connection site.)

[0155] (Supplementary Note 3) The above R 1 is selected from any one of the following structures, and the biphenyl derivative according to Supplementary Note 2 is characterized thereby. [Chemical formula] (“*” indicates the connecting site.)

[0156] (Supplementary Note 4) The A ring is selected from any one of the following structures, and the biphenyl derivative according to Supplementary Note 1 is characterized thereby. [Chemical formula] (X is CR 3 R 4 or NR 5 and R 2 ~R 5 are each independently selected from -H, -CH3 or -NH2, “*” indicates the connecting site.)

[0157] (Supplementary Note 5) The A ring is selected from any one of the following structures, and the biphenyl derivative according to Supplementary Note 3 is characterized thereby. [Chemical formula] (“*” indicates the connecting site.)

[0158] (Supplementary Note 6) The A ring is selected from any one of the following structures, and the biphenyl derivative according to Supplementary Note 3 is characterized thereby. [Chemical formula] (“*” indicates the connecting site.)

[0159] (Supplementary Note 7) It has a structure represented by any of Formulae I-1 to I-18, or is a pharmaceutically acceptable salt, ester, amide, solvate, active metabolite, polymorph, isotope-labeled form, isomer or prodrug of a structure represented by any of Formulae I-1 to I-18, and is a biphenyl derivative according to Appendix 1. [Chemical formula]

[0160] (Appendix 8) A pharmaceutical composition comprising a biphenyl derivative according to any one of Appendices 1 to 7 and at least one pharmaceutically acceptable carrier.

[0161] (Appendix 9) Use of a biphenyl derivative according to any one of Appendices 1 to 7 or a pharmaceutical composition according to Appendix 8 in cell reprogramming.

[0162] (Appendix 10) A method for cell reprogramming, comprising the step of contacting and treating cells with a biphenyl derivative according to any one of Appendices 1 to 7 or a pharmaceutical composition according to Appendix 8.

[0163] (Appendix 11) The method according to Appendix 10, wherein the concentration of the biphenyl derivative in the contact treatment is 1 μM to 50 μM.

[0164] (Appendix 12) Use of a biphenyl derivative according to any one of Appendices 1 to 7 or a pharmaceutical composition according to Appendix 8 in kinase inhibition, wherein the kinase comprises one or more of the TK kinase family, the STE kinase family and the CMGC kinase family.

[0165] (Appendix 13) The use according to Appendix 12, wherein the kinase comprises one or more of PDGFRα, TNIK, CLK4 and JNK1.

[0166] (Note 14) The process includes the step of contacting the kinase with any one of the biphenyl derivatives described in Appendix 1 to 7 or the pharmaceutical composition described in Appendix 8, A kinase inhibition method wherein the kinase comprises one or more of the TK kinase family, the STE kinase family, and the CMGC kinase family.

[0167] (Note 15) The method according to Appendix 14, characterized in that the kinase comprises one or more of PDGFRα, TNIK, CLK4, and JNK1.

Claims

1. A biphenyl derivative having the structure represented by formula I, or a pharmaceutically acceptable salt, ester, amide, solvate, active metabolite, polymorph, isotope-labeled, isomer, or prodrug of the structure represented by formula I. 【Chemistry 1】 (In the formula, ring A is a methyl group or amino group substituted or unsubstituted five-membered ring, and the ring atoms of ring A contain one or two nitrogen atoms, R １ (A hydrogen bond is a donor or acceptor, and its structure contains one or more of the following: an amino group, an imino group, a hydroxyl group, and an ether bond.)

2. The aforementioned R １ The biphenyl derivative according to claim 1, characterized in that it is selected from any one of the following structures. 【Chemistry 2】 (The asterisk "*" indicates a connecting point.)

3. The aforementioned R １ The biphenyl derivative according to claim 2, characterized in that it is selected from any one of the following structures. 【Transformation 3】 (The asterisk "*" indicates a connecting point.)

4. The biphenyl derivative according to claim 1, characterized in that the A ring is selected from any one of the following structures. 【Chemistry 4】 (X is CR ３ R ４ or NR ５ and R ２ ~ R ５ are each independently selected from -H, -CH ３ or -NH ２ and (* indicates a connecting point.)

5. The biphenyl derivative according to claim 3, characterized in that the A ring is selected from any one of the following structures. 【Transformation 5】 (The asterisk "*" indicates a connecting point.)

6. The biphenyl derivative according to claim 3, characterized in that the A ring is selected from any one of the following structures. 【Transformation 6】 (The asterisk "*" indicates a connecting point.)

7. The biphenyl derivative according to claim 1, characterized in that it has a structure represented by any of formulas I-1 to I-18, or is a pharmaceutically acceptable salt, ester, amide, solvate, active metabolite, polymorph, isotope-labeled, isomer, or prodrug of a structure represented by any of formulas I-1 to I-18. 【Transformation 7】

8. A pharmaceutical composition comprising a biphenyl derivative according to any one of claims 1 to 7 and at least one pharmaceutically acceptable carrier.

9. Use of the biphenyl derivative according to any one of claims 1 to 7 or the pharmaceutical composition according to claim 8 in cell reprogramming.

10. A cell reprogramming method comprising the step of treating cells by contacting them with a biphenyl derivative according to any one of claims 1 to 7 or a pharmaceutical composition according to claim 8.

11. The method according to claim 10, characterized in that the concentration of the biphenyl derivative in the contact treatment is 1 μM to 50 μM.

12. The use of a biphenyl derivative according to any one of claims 1 to 7 or a pharmaceutical composition according to claim 8 in kinase inhibition, The use of the kinase includes one or more of the TK kinase family, STE kinase family, and CMGC kinase family.

13. The use according to claim 12, characterized in that the kinase comprises one or more of PDGFRα, TNIK, CLK4, and JNK1.

14. The process includes the step of contacting the kinase with the biphenyl derivative described in any one of claims 1 to 7 or the pharmaceutical composition described in claim 8, A kinase inhibition method wherein the kinase comprises one or more of the TK kinase family, the STE kinase family, and the CMGC kinase family.

15. The method according to claim 14, characterized in that the kinase comprises one or more of PDGFRα, TNIK, CLK4, and JNK1.

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