KR20250132936A - The new small molecular compounds having IRP2 inhibition activity - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating cancer, comprising a benzenesulfonamide-substituted heterobicyclic derivative, which is an IRP2 disruptor, as an active ingredient, and can provide a benzenesulfonamide-substituted heterobicyclic derivative compound that kills solid cancer cells, especially colon cancer cells, by interfering with IRP2 binding to IRE and inducing reprogramming of iron metabolism in cancer cells.

Description

The new small molecular compounds having IRP2 inhibition activity

The present invention relates to a benzenesulfonamide substituted heterobicyclic derivative which is an IRP2 disruptor, a method for preparing the same, and a pharmaceutical use thereof.

The present invention relates to a pharmaceutical composition for preventing or treating cancer of a benzenesulfonamide-substituted heterobicyclic derivative, which is an IRP2 disruptor.

The present invention relates to a pharmaceutical composition for preventing or treating colon cancer of a benzenesulfonamide-substituted heterobicyclic derivative, which is an IRP2 disruptor.

Iron is a biologically essential component for DNA synthesis, mitochondrial respiration, and cell proliferation. Iron homeostasis is involved in redox activity, mitochondrial function, cell growth, and apoptosis (Miyazawa et al., 2019; Zhang et al., 2020). Iron metabolism dysregulation is common in cancer cells, particularly because cancer cells require a relatively higher iron requirement to sustain cell growth compared to normal cells.

In all mammals, iron regulatory proteins 1 and 2 (IRP1 and 2) control cellular iron homeostasis through binding to iron-responsive elements (IREs) in the 5' or 3' untranslated regions (UTRs) of selected mRNAs (Jiao et al., 2019; Wang et al., 2020). Typically, IRP binding to IREs in the 5' UTR of mRNAs represses the translation of proteins including ferritin H (FTH) and ferroportin (FPN) (Li et al., 2019; Martelli et al., 2015), whereas IREs in the 3' UTR of mRNAs activate protein stability, including transferrin receptor 1 (TfR1) and divalent metal ion transporter 1 (DMT1) (Bellelli et al., 2016; Wallander et al., 2008).

IRP1 and IRP2 share a high degree of nucleotide sequence homology, are differentially expressed depending on cellular iron availability, and are crucial for iron metabolism reprogramming (Miyazawa et al., 2019). Of note, IRP1 is a ubiquitously expressed protein with a 4Fe-4S cluster that disallows IRP1 binding to IREs, whereas IRP2 is a selectively expressed protein lacking the iron-sulfur cluster and reported to be the dominant IRE-binding protein (Moroishi et al., 2011). In cellular iron overload, IRP1 activates 4Fe-4S binding, whereas IRP2 is degraded by F-box and Leucine Rich Repeat Protein 5 (FBXL5), an E3 ubiquitin ligase (Muto et al., 2017; Zumbrennen-Bullough et al., 2014).

Colorectal cancer is the second most lethal cancer worldwide, following lung cancer. Compared to other cancer types, colorectal cancer has a lower response rate to standard therapies and exhibits resistance to targeted therapies. For example, epidermal growth factor receptor (EGFR) inhibitors are effective in patients with KRAS wild-type tumors (Missiaglia et al., 2014). Furthermore, while promising, newly developed immunotherapies ultimately affect less than 5% of all metastatic colorectal cancer patients with high microsatellite instability (MSI-H), which limits treatment development (Andre et al., 2020). Therefore, despite the increasing incidence of colorectal cancer, progress in the development of anticancer drugs for colorectal cancer remains stagnant, with a 5-year survival rate of only 15% for patients with stage IV colorectal cancer (Siegel et al., 2019). Despite previous studies supporting the importance of iron overload and its direct consequences in colorectal cancer, no treatment exists that reprograms iron metabolism in cancer cells, other than iron chelators. Therefore, the inventors of the present invention hypothesized that by utilizing the differences in iron metabolism between cancer and normal cells, IRP2 can be targeted to specifically inhibit cancer cell proliferation, leading to the development of an effective cancer treatment.

In this study, we identified a novel small molecule inhibitor that blocks IRP2 binding to IRE through comprehensive quantum analysis, pharmacophore modeling, and molecular docking. We found that genetic or pharmacological inhibition of IRP2 disrupts cellular iron homeostasis and inhibits colon cancer cell growth in various model systems, including patient-derived cell lines, organoids, and tumor xenografts. Furthermore, we elucidated the mechanism underlying the therapeutic effects of Compound 1, the first IRP2 inhibitor, highlighting the relevance of targeting IRP2 in colorectal cancer.

Andre, T. et al., (2020). Pembrolizumab in Microsatellite-Instability-High Advanced Colorectal Cancer. N Engl J Med 383, 2207-2218. Bellelli, R. et al., (2016). NCOA4 Deficiency Impairs Systemic Iron Homeostasis. Cell Rep 14, 411-421. Jiao, Q. et al., (2019). Oxidative Stress Regulated Iron Regulatory Protein IRP2 Through FBXL5-Mediated Ubiquitination-Proteasome Way in SH-SY5Y Cells. Front Neurosci 13, 20. Li, H. et al., (2019). Iron regulatory protein 2 modulates the switch from aerobic glycolysis to oxidative phosphorylation in mouse embryonic fibroblasts. Proc Natl Acad Sci U S A 116, 9871-9876. Martelli, A. et al., (2015). Iron regulatory protein 1 sustains mitochondrial iron loading and function in frataxin deficiency. Cell Metab 21, 311-323. Missiaglia, E. et al., (2014). Distal and proximal colon cancers differ in terms of molecular, pathological, and clinical features. Ann Oncol 25, 1995-2001. Miyazawa, M. et al., (2019). Perturbation of Iron Metabolism by Cisplatin through Inhibition of Iron Regulatory Protein 2. Cell Chem Biol 26, 85-97 e84. Moroishi, T. et al., (2011). The FBXL5-IRP2 axis is integral to control of iron metabolism in vivo. Cell Metab 14, 339-351. Muto, Y. et al., (2017). Essential role of FBXL5-mediated cellular iron homeostasis in maintenance of hematopoietic stem cells. Nat Commun 8, 16114. Selezneva, A.I. et al., (2013). Nucleotide-specific recognition of iron-responsive elements by iron regulatory protein 1. J Mol Biol 425, 3301-3310. Siegel, R.L. et al., (2019). Cancer statistics, 2019. CA: a cancer journal for clinicians 69, 7-34. Wallander, M.L. et al., (2008). Iron-independent phosphorylation of iron regulatory protein 2 regulates ferritin during the cell cycle. J Biol Chem 283, 23589-23598. Wang, H. et al., (2020). FBXL5 Regulates IRP2 Stability in Iron Homeostasis via an Oxygen-Responsive [2Fe2S] Cluster. Mol Cell. Zhang, J. et al., (2020). Mdm2 is a target and mediator of IRP2 in cell growth control. FASEB J 34, 2301-2311. Zumbrenn-Bullough, K.B. et al., (2014). Abnormal brain iron metabolism in Irp2 deficient mice is associated with mild neurological and behavioral impairments. PLoS One 9, e98072.

The purpose of the present invention is to provide a pharmaceutical composition for preventing or treating colon cancer, which comprises as an active ingredient a benzenesulfonamide-substituted heterobicyclic derivative that kills colon cancer cells by inducing reprogramming of iron metabolism in cancer cells by interfering with IRP2 binding to IRE.

The present invention relates to a pharmaceutical composition for preventing or treating colon cancer, comprising a compound represented by the following chemical formula 1, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

[Chemical Formula 1]

In the above chemical formula 1,

R ₁ is substituted with one or more substituents selected from the group consisting of hydrogen, halogen, hydroxy, amino, amide, cyano, halo C ₁ -C ₆ alkyl, NO ₂ , C _{1 -C 6} alkyl _, C ₁ -C ₁₀ alkoxy, acetyl, acetamido, C ₁ -C ₆ alkylamino, di(C ₁ -C ₆ ) alkylamino, C ₁ -C ₆ alkylcarbonylamino, C ₅ -C ₁₀ aryl, C ₅ -C ₁₀ heteroaryl, C 4 -C 10 cycloalkyl and C ₄ -C ₁₀ heterocycloalkyl;

R ₂ is substituted with one or more substituents selected from the group consisting of hydrogen, C ₁ -C ₆ alkyl, halo C ₁ -C ₆ alkyl, C ₁ -C ₁₀ alkoxy, acetyl, C _{1 -C 6} alkylamino, di(C ₁ -C _{6 )} alkylamino, di(C 1 -C ₆₎ alkylamino (C ₁ -C 6) alkyl, phenyl, and SEM(=2-(trimethylsilyl)ethoxymethyl);

R ₃ is independently hydrogen, halogen or C ₁ -C ₁₀ alkoxy;

R ₄ is substituted with one or more substituents selected from the group consisting of substituted or unsubstituted (C ₅ -C ₁₀ )aryl-amino, substituted or unsubstituted (C ₅ -C ₁₀ )heteroaryl-amino, substituted or unsubstituted C ₅ -C ₁₀ heteroaryl, and substituted or unsubstituted C ₄ -C ₁₀ heterocycloalkyl,

The above substituted (C ₅ -C ₁₀ ) aryl-amino, (C ₅ -C ₁₀ ) heteroaryl-amino, C ₅ -C ₁₀ heteroaryl and C ₄ -C ₁₀ heterocycloalkyl are substituted with one or more substituents independently selected from the group consisting of hydrogen, halogen, hydroxy, amide, halo C ₁ -C ₆ alkyl, cyano, NO ₂ , C ₁ -C ₆ alkyl, acetyl, carbamoyl, substituted or unsubstituted C ₁ -C ₁₀ alkoxy, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, and substituted or unsubstituted C ₄ -C ₁₀ heterocycloalkyl,

The above substituted C ₁ -C ₁₀ alkoxy, C ₃ -C ₁₀ cycloalkyl or C ₄ -C ₁₀ heterocycloalkyl is substituted with one or more substituents selected from the group consisting of hydrogen, halogen, hydroxy, amide, halo C ₁ -C ₆ alkyl, cyano, NO ₂ , C ₁ -C ₆ alkyl, and C ₁ -C ₁₀ alkoxy;

A is CR ₁ , CH or N;

is a single bond or double bond;

n is an integer from 0 to 3;

The present invention relates to a pharmaceutical composition for preventing or treating colon cancer, comprising a compound represented by the above chemical formula 1, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

In the above chemical formula 1,

R ₁ is hydrogen, halogen, hydroxy, amino, amide, CN, halo C ₁ -C ₆ alkyl, NO ₂ , C ₁ -C ₆ alkyl, C ₁ -C ₁₀ alkoxy, acetyl, acetamido, C ₁ -C ₆ alkylamino, di(C ₁ -C ₆ ) Substituted with one or more substituents selected from the group consisting of alkylamino, C ₁ -C ₆ alkylcarbonylamino, phenyl, thiophenyl, and morpholinyl;

R ₂ is substituted with one or more substituents selected from the group consisting of hydrogen, halogen, hydroxy, amide, CN, halo C ₁ -C ₆ alkyl, NO ₂ , C _{1 -C 6} alkyl, C ₁ -C ₁₀ alkoxy, acetyl, C ₁ -C ₆ alkylamino, di(C ₁ -C _{6 )} alkylamino, di(C ₁ -C _{6) alkylamino(C 1 -C 6)} alkyl, phenyl, and SEM(=2-(trimethylsilyl)ethoxymethyl);

R ₃ is independently hydrogen, halogen or C ₁ -C ₁₀ alkoxy;

R ₄ is substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted and substituted or unsubstituted is substituted with one or more substituents selected from the group consisting of,

The above substituted , , , , and is substituted with one or more substituents independently selected from the group consisting of hydrogen, halogen, hydroxy, amide, halo C ₁ -C ₆ alkyl, CN, NO ₂ , C ₁ -C ₆ alkyl, acetyl, carbamoyl, substituted or unsubstituted C ₁ -C ₁₀ alkoxy, substituted or unsubstituted C ₄ -C ₁₀ cycloalkyl, and substituted or unsubstituted C ₄ -C ₁₀ heterocycloalkyl,

The above substituted C ₁ -C ₁₀ alkoxy, C ₃ -C ₁₀ cycloalkyl or C ₄ -C ₁₀ heterocycloalkyl is substituted with one or more substituents selected from the group consisting of hydrogen, halogen, hydroxy, amide, halo C ₁ -C ₆ alkyl, CN, NO ₂ , C ₁ -C ₆ alkyl, and C ₁ -C ₁₀ alkoxy;

A is CR ₁ , CH or N;

is a single bond or double bond;

n is an integer from 0 to 3;

The present invention relates to a pharmaceutical composition for preventing or treating colon cancer, comprising a compound represented by the following chemical formula 2, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

[Chemical Formula 2]

In the above chemical formula 2,

R ₁ is substituted with one or more substituents selected from the _group consisting of hydrogen, halogen, hydroxy, amino, amide, CN, halo C ₁ -C ₆ alkyl, NO ₂ , C _{1 -C 6} alkyl, C ₁ -C ₁₀ alkoxy, acetyl, acetamido, C 1 -C 6 alkylamino, di(C ₁ -C ₆₎ alkylamino, C ₁ -C ₆ alkylcarbonylamino, phenyl, thiophenyl, and morpholinyl;

R ₂ is substituted with one or more substituents selected from the group consisting of hydrogen, halogen, hydroxy, amide, CN, halo C ₁ -C ₆ alkyl, NO ₂ , C _{1 -C 6} alkyl, C ₁ -C ₁₀ alkoxy, acetyl, C ₁ -C ₆ alkylamino, di(C ₁ -C _{6 )} alkylamino, di(C ₁ -C _{6) alkylamino (C 1 -C 6)} alkyl, phenyl, and SEM(=2-(trimethylsilyl)ethoxymethyl);

R ₃ is independently hydrogen, halogen or C ₁ -C ₁₀ alkoxy;

R ₄ is substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted and substituted or unsubstituted is substituted with one or more substituents selected from the group consisting of,

The above substituted , , , , and is substituted with one or more substituents independently selected from the group consisting of hydrogen, halogen, hydroxy, amide, halo C ₁ -C ₆ alkyl, CN, NO ₂ , C ₁ -C ₆ alkyl, acetyl, carbamoyl, substituted or unsubstituted C ₁ -C ₁₀ alkoxy, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted C ₄ -C ₁₀ heterocycloalkyl,

The above substituted C ₁ -C ₁₀ alkoxy, C ₃ -C ₁₀ cycloalkyl or C ₄ -C ₁₀ heterocycloalkyl is substituted with one or more substituents selected from the group consisting of hydrogen, halogen, hydroxy, amide, halo C1-C6 alkyl, CN, NO ₂ , C ₁ -C ₆ alkyl, C ₁ -C ₁₀ alkoxy;

n is an integer from 0 to 3;

The present invention relates to a pharmaceutical composition for preventing or treating colon cancer, comprising a compound represented by the above chemical formula 2, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

In the above chemical formula 2,

R ₁ is substituted with one or more substituents selected from the group consisting of hydrogen, halo C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl, acetyl, acetamido, phenyl, thiophenyl, and morpholinyl;

R ₂ is substituted with one or more substituents selected from the group consisting of hydrogen, C ₁ -C ₆ alkyl and acetyl;

R ₃ is independently hydrogen, halogen or C ₁ -C ₆ alkoxy;

R ₄ is substituted or unsubstituted and substituted or unsubstituted is substituted with one or more substituents selected from the group consisting of,

The above substituted and In which at least one hydrogen in the substituent is independently substituted with at least one substituent selected from the group consisting of hydrogen, halogen, halo C ₁ -C ₆ alkyl, CN and C ₁ -C ₆ alkyl,

n is an integer from 0 to 3;

The present invention relates to a pharmaceutical composition for preventing or treating colon cancer, comprising a compound represented by the following chemical formula 3, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

[Chemical Formula 3]

In the above chemical formula 3,

R ₁ is hydrogen, halogen, hydroxy, amino, amide, CN, halo C ₁ -C ₆ alkyl, NO ₂ , C ₁ -C ₆ alkyl, C ₁ -C ₁₀ alkoxy, acetyl, acetamido, C ₁ -C ₆ alkylamino, di(C ₁ -C ₆ ) Substituted with one or more substituents selected from the group consisting of alkylamino, C ₁ -C ₆ alkylcarbonylamino, phenyl, thiophenyl, and morpholinyl;

R ₂ is substituted with one or more substituents selected from the group consisting of hydrogen, halogen, hydroxy, amide, CN, halo C ₁ -C ₆ alkyl, NO ₂ , C _{1 -C 6} alkyl, C ₁ -C ₁₀ alkoxy, acetyl, C ₁ -C ₆ alkylamino, di(C ₁ -C _{6 )} alkylamino, di(C ₁ -C _{6) alkylamino(C 1 -C 6)} alkyl, phenyl, and SEM(=2-(trimethylsilyl)ethoxymethyl);

R ₃ is independently hydrogen, halogen or C ₁ -C ₁₀ alkoxy;

R ₄ is substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted and substituted or unsubstituted is substituted with one or more substituents selected from the group consisting of,

The above substituted , , , , and is substituted with one or more substituents independently selected from the group consisting of hydrogen, halogen, hydroxy, amide, halo C ₁ -C ₆ alkyl, CN, NO ₂ , C ₁ -C ₆ alkyl, acetyl, carbamoyl, substituted or unsubstituted C ₁ -C ₁₀ alkoxy, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted C ₄ -C ₁₀ heterocycloalkyl,

The above substituted C ₁ -C ₁₀ alkoxy, C ₃ -C ₁₀ cycloalkyl or C ₄ -C ₁₀ heterocycloalkyl is substituted with one or more substituents selected from the group consisting of hydrogen, halogen, hydroxy, amide, halo C ₁ -C ₆ alkyl, CN, NO ₂ , C ₁ -C ₆ alkyl, C ₁ -C ₁₀ alkoxy;

n is an integer from 0 to 3;

The present invention relates to a pharmaceutical composition for preventing or treating colon cancer, comprising a compound represented by the above chemical formula 3, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

In the above chemical formula 3,

R ₁ is substituted with one or more substituents selected from the group consisting of hydrogen, amino, amide, halo C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl, acetyl, acetamido, phenyl, thiophenyl, and morpholinyl;

R ₂ is substituted with one or more substituents selected from the group consisting of hydrogen, halo C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl, and acetyl;

R ₃ is independently hydrogen, halogen or C ₁ -C ₆ alkoxy;

R ₄ is substituted or unsubstituted and substituted or unsubstituted is substituted with one or more substituents selected from the group consisting of,

The above substituted and wherein at least one hydrogen in the substituent is independently substituted with at least one substituent selected from the group consisting of hydrogen, halogen, halo C ₁ -C ₆ alkyl, CN, C ₁ -C ₆ alkyl and acetyl;

n is an integer from 0 to 3;

The present invention relates to a pharmaceutical composition for preventing or treating colon cancer, comprising a compound represented by the following chemical formula 4, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

[Chemical Formula 4]

In the above chemical formula 4,

R ₁ is hydrogen, halogen, hydroxy, amino, amide, CN, halo C ₁ -C ₆ alkyl, NO ₂ , C ₁ -C ₆ alkyl, C ₁ -C ₁₀ alkoxy, acetyl, acetamido, C ₁ -C ₆ alkylamino, di(C ₁ -C ₆ ) Substituted with one or more substituents selected from the group consisting of alkylamino, C ₁ -C ₆ alkylcarbonylamino, phenyl, thiophenyl, and morpholinyl;

R ₂ is substituted with one or more substituents selected from the group consisting of hydrogen, halogen, hydroxy, amide, CN, halo C ₁ -C ₆ alkyl, NO ₂ , C _{1 -C 6} alkyl, C ₁ -C ₁₀ alkoxy, acetyl, C ₁ -C ₆ alkylamino, di(C ₁ -C ₆ ) alkylamino, di(C ₁ -C _{6 ) alkylamino (C 1 -C 6)} alkyl, phenyl, and SEM(=2-(trimethylsilyl)ethoxymethyl);

R ₃ is independently hydrogen, halogen or C ₁ -C ₁₀ alkoxy;

R ₄ is substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted and substituted or unsubstituted is substituted with one or more substituents selected from the group consisting of,

The above substituted , , , , and is substituted with one or more substituents independently selected from the group consisting of hydrogen, halogen, hydroxy, amide, halo C ₁ -C ₆ alkyl, CN, NO ₂ , C ₁ -C ₆ alkyl, acetyl, carbamoyl, substituted or unsubstituted C ₁ -C ₁₀ alkoxy, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted C ₄ -C ₁₀ heterocycloalkyl,

The above substituted C ₁ -C ₁₀ alkoxy, C ₃ -C ₁₀ cycloalkyl or C ₄ -C ₁₀ heterocycloalkyl is substituted with one or more substituents selected from the group consisting of hydrogen, halogen, hydroxy, amide, halo C ₁ -C ₆ alkyl, CN, NO ₂ , C ₁ -C ₆ alkyl, C ₁ -C ₁₀ alkoxy;

n is an integer from 0 to 3;

The present invention relates to a pharmaceutical composition for preventing or treating colon cancer, comprising a compound represented by the above chemical formula 4, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

In the above chemical formula 4,

R ₁ is substituted with one or more substituents selected from the group consisting of hydrogen, amino, amide, halo C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl, acetyl, acetamido, phenyl, thiophenyl, and morpholinyl;

R ₂ is substituted with one or more substituents selected from the group consisting of hydrogen, halo C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl, and acetyl;

R ₃ is independently hydrogen, halogen or C ₁ -C ₆ alkoxy;

R ₄ is substituted or unsubstituted and substituted or unsubstituted is substituted with one or more substituents selected from the group consisting of,

The above substituted and wherein at least one hydrogen in the substituent is independently substituted with at least one substituent selected from the group consisting of hydrogen, halogen, halo C ₁ -C ₆ alkyl, CN, C ₁ -C ₆ alkyl and acetyl;

n is an integer from 0 to 3;

The present invention relates to a pharmaceutical composition for preventing or treating colon cancer, comprising a compound represented by the following chemical formula 5, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

[Chemical Formula 5]

In the above chemical formula 5,

R ₁ is substituted with one or more substituents selected from the _group consisting of hydrogen, halogen, hydroxy, amino, amide, CN, halo C ₁ -C ₆ alkyl, NO ₂ , C _{1 -C 6} alkyl, C ₁ -C ₁₀ alkoxy, acetyl, acetamido, C 1 -C 6 alkylamino, di(C ₁ -C ₆₎ alkylamino, C ₁ -C ₆ alkylcarbonylamino, phenyl, thiophenyl, and morpholinyl;

R ₂ is substituted with one or more substituents selected from the group consisting of hydrogen, halogen, hydroxy, amide, CN, halo C ₁ -C ₆ alkyl, NO ₂ , C _{1 -C 6} alkyl, C ₁ -C ₁₀ alkoxy, acetyl, C ₁ -C ₆ alkylamino, di(C ₁ -C _{6 )} alkylamino, di(C ₁ -C _{6) alkylamino (C 1 -C 6)} alkyl, phenyl, and SEM(=2-(trimethylsilyl)ethoxymethyl);

R ₃ is independently hydrogen, halogen or C ₁ -C ₆ alkoxy;

R ₄ is substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted and substituted or unsubstituted is substituted with one or more substituents selected from the group consisting of,

The above substituted , , , , and is substituted with one or more substituents independently selected from the group consisting of hydrogen, halogen, hydroxy, amide, halo C ₁ -C ₆ alkyl, CN, NO ₂ , C ₁ -C ₆ alkyl, acetyl, carbamoyl, substituted or unsubstituted C ₁ -C ₁₀ alkoxy, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted C ₄ -C ₁₀ heterocycloalkyl,

The above substituted C ₁ -C ₁₀ alkoxy, C ₃ -C ₁₀ cycloalkyl or C ₄ -C ₁₀ heterocycloalkyl is substituted with one or more substituents selected from the group consisting of hydrogen, halogen, hydroxy, amide, halo C ₁ -C ₆ alkyl, CN, NO ₂ , C ₁ -C ₆ alkyl, C ₁ -C ₁₀ alkoxy;

n is an integer from 0 to 3;

The present invention relates to a pharmaceutical composition for preventing or treating colon cancer, comprising a compound represented by the above chemical formula 5, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

In the above chemical formula 5,

R ₁ is substituted with one or more substituents selected from the group consisting of hydrogen, amino, amide, halo C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl, acetyl, acetamido, phenyl, thiophenyl, and morpholinyl;

R ₂ is substituted with one or more substituents selected from the group consisting of hydrogen, halo C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl, and acetyl;

R ₃ is independently hydrogen, halogen or C ₁ -C ₆ alkoxy;

R ₄ is substituted or unsubstituted and substituted or unsubstituted is substituted with one or more substituents selected from the group consisting of,

The above substituted and wherein at least one hydrogen in the substituent is independently substituted with at least one substituent selected from the group consisting of hydrogen, halogen, halo C ₁ -C ₆ alkyl, CN, C ₁ -C ₆ alkyl and acetyl;

n is an integer from 0 to 3;

To give a more specific example of the compound of the present invention,

4-Methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide (Compound 1);

4-Methoxy-N-phenyl-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 2);

4-Methoxy-N-(3-methoxyphenyl)-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 3);

3-((4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)phenyl)sulfonamido)benzamide (compound 4);

4-Methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(3-(trifluoromethyl)phenyl)benzenesulfonamide (Compound 5);

N-(3-Cyanophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 6);

N-(3-Fluorophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 7);

N-(4-Fluorophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 8);

N-(2-Fluorophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 9);

N-(3-chlorophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 10);

N-(4-chlorophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 11);

N-(2-chlorophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 12);

N-(2-Cyclopropylphenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 13);

N-(4-hydroxyphenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 14);

N-(3-(2-hydroxyethoxy)phenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 15);

N-(4-(2-hydroxyethoxy)phenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 16);

N-(2-(2-hydroxyethoxy)phenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 17);

4-Methoxy-N-(4-(4-methylpiperazin-1-yl)phenyl)-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 18);

4-Methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(pyridin-3-yl)benzenesulfonamide (Compound 19);

N-(3-fluorophenyl)-3-(4-oxo-1,4-dihydroquinolin-6-yl)benzenesulfonamide (Compound 20);

5-(5-((1H-Pyrazol-1-yl)sulfonyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridine (Compound 21);

5-(2-methoxy-5-((4-methylpiperazin-1-yl)sulfonyl)phenyl)-1H-pyrazolo[3,4-b]pyridine (Compound 22);

4-((4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)phenyl)sulfonyl)morpholine (Compound 23);

N-(3,5-difluorophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 24);

N-(3-chloro-5-fluorophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 25);

4-Methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzamide (Compound 26);

N-(3-Fluorophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzamide (Compound 27);

N-Phenyl-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 28);

3-(1H-Pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide (Compound 29);

N-(3-Fluorophenyl)-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 30);

N-(3-chlorophenyl)-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 31);

3-(1H-Pyrazolo[3,4-b]pyridin-5-yl)-N-(o-tolyl)benzenesulfonamide (Compound 32);

4-Methoxy-3-(3-methyl-1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide (Compound 33);

4-Methoxy-3-(1-phenyl-1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide (Compound 34);

N-(5-(2-methoxy-5-(N-(m-tolyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 35);

N-(5-(4-fluoro-3-(N-(m-tolyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 36);

N-(5-(2-Fluoro-5-(N-(m-tolyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 37);

N-(5-(3-fluoro-5-(N-(m-tolyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 38);

N-(5-(2,4-difluoro-5-(N-(m-tolyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 39);

N-(5-(2-Fluoro-3-(N-(m-tolyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 40);

N-(5-(2-methoxy-5-(N-(p-tolyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 41);

N-(5-(2-methoxy-5-(N-(o-tolyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 42);

N-(5-(2-methoxy-5-(N-phenylsulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 43);

N-(5-(5-(N-(3-fluorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 44);

N-(5-(4-fluoro-3-(N-(3-fluorophenyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 45);

N-(5-(2-Fluoro-5-(N-(3-fluorophenyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 46);

N-(5-(3-fluoro-5-(3-fluorophenyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 47);

N-(5-(2,4-difluoro-5-(N-(3-fluorophenyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 48);

N-(5-(2-Fluoro-3-(N-(3-fluorophenyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 49);

N-(5-(5-(N-(4-fluorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 50);

N-(5-(5-(N-(2-fluorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 51);

N-(5-(5-(N-(3-chlorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 52);

N-(5-(5-(N-(4-chlorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 53);

N-(5-(5-(N-(2-chlorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 54);

N-(5-(5-(N-(3-hydroxyphenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 55);

N-(5-(5-(N-(4-hydroxyphenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 56);

N-(5-(2-methoxy-5-(N-(pyridin-3-yl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 57);

N-(5-(2-methoxy-5-(N-(pyridin-2-yl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 58);

N-(5-(2-methoxy-5-(morpholinosulfonyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 59);

N-(5-(5-(N-(3,5-difluorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 60);

N-(5-(5-(N-(3-chloro-5-fluorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 61);

N-(3-Fluorophenyl)-4-methoxy-3-(3-phenyl-1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 62);

N-(3-Fluorophenyl)-4-methoxy-3-(3-(thiophen-3-yl)-1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 63);

4-Methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide (Compound 64);

4-Methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(p-tolyl)benzenesulfonamide (Compound 65);

4-Methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(o-tolyl)benzenesulfonamide (Compound 66);

4-Methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)-N-phenylbenzenesulfonamide (Compound 67);

N-(3-Fluorophenyl)-4-methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 68);

N-(2-Fluorophenyl)-4-methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 69);

N-(2-Chlorophenyl)-4-methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 70);

N-(3-Hydroxyphenyl)-4-methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 71);

4-((4-methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)phenyl)sulfonyl)morpholine (Compound 72);

4-Methoxy-3-(1-phenyl-1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide (Compound 73);

3-(1-(2-(Dimethylamino)ethyl)-1H-pyrazolo[3,4-b]pyridin-5-yl)-4-methoxy-N-(m-tolyl)benzenesulfonamide (Compound 74);

4-Methoxy-3-(1-methyl-1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide (Compound 75);

3-(1-Acetyl-1H-pyrazolo[3,4-b]pyridin-5-yl)-4-methoxy-N-(m-tolyl)benzenesulfonamide (Compound 76);

N-((3-(1-Acetyl-1H-pyrazolo[3,4-b]pyridin-5-yl)-4-methoxyphenyl)sulfonyl)-N-(m-tolyl)acetamide (Compound 77);

4-((3-(1H-indazol-5-yl)-4-methoxyphenyl)sulfonyl)morpholine (Compound 78);

3-(1H-indazol-5-yl)-4-methoxy-N-(m-tolyl)benzenesulfonamide (Compound 79);

N-(3-Fluorophenyl)-3-(1H-indazol-5-yl)-4-methoxybenzenesulfonamide (Compound 80);

2,4-Difluoro-N-(3-fluorophenyl)-5-(1H-indazol-5-yl)benzenesulfonamide (Compound 81);

N-(5-(5-(N-(3-fluorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-indazol-3-yl)acetamide (Compound 82);

N-(5-(2,4-difluoro-5-(N-(3-fluorophenyl)sulfamoyl)phenyl)-1H-indazol-3-yl)acetamide (Compound 83);

3-(1-Acetyl-3-amino-1H-indazol-5-yl)-N-(3-fluorophenyl)-4-methoxybenzenesulfonamide (Compound 84);

5-(1-Acetyl-3-amino-1H-indazol-5-yl)-2,4-difluoro-N-(3-fluorophenyl)benzenesulfonamide (Compound 85);

4-((4-methoxy-3-(1H-pyrrolo[2,3-b]pyridin-5-yl)phenyl)sulfonyl)morpholine (Compound 86);

4-Methoxy-3-(1H-pyrrolo[2,3-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide (Compound 87);

N-(3-Fluorophenyl)-4-methoxy-3-(1H-pyrrolo[2,3-b]pyridin-5-yl)benzenesulfonamide (Compound 88);

4-Methoxy-3-(3-morpholino-1H-pyrrolo[2,3-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide (Compound 90);

4-Methoxy-3-(3-morpholino-1H-pyrrolo[2,3-b]pyridin-5-yl)-N-(o-tolyl)benzenesulfonamide (Compound 91);

4-Methoxy-3-(3-morpholino-1H-pyrrolo[2,3-b]pyridin-5-yl)-N-phenylbenzenesulfonamide (Compound 92);

N-(2-Fluorophenyl)-4-methoxy-3-(3-morpholino-1H-pyrrolo[2,3-b]pyridin-5-yl)benzenesulfonamide (Compound 93);

N-(2-Chlorophenyl)-4-methoxy-3-(3-morpholino-1H-pyrrolo[2,3-b]pyridin-5-yl)benzenesulfonamide (Compound 94);

N-(5-(2-methoxy-5-(N-phenylsulfamoyl)phenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)acetamide (Compound 95);

N-(5-(2,4-difluoro-5-(N-(3-fluorophenyl)sulfamoyl)phenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)acetamide (Compound 96);

3-(1H-Indol-5-yl)-4-methoxy-N-(m-tolyl)benzenesulfonamide (Compound 97);

N-(3-Fluorophenyl)-3-(1H-indol-5-yl)-4-methoxybenzenesulfonamide (Compound 98);

4-Methoxy-N-(m-tolyl)-3-(3-((2-(trimethylsilyl)ethoxy)methyl)-3H-imidazo[4,5-b]pyridin-6-yl)benzenesulfonamide (Compound 99);

3-(3H-Imidazo[4,5-b]pyridin-6-yl)-4-methoxy-N-(m-tolyl)benzenesulfonamide (Compound 100);

N-(3-Fluorophenyl)-4-methoxy-3-(3-((2-(trimethylsilyl)ethoxy)methyl)-3H-imidazo[4,5-b]pyridin-6-yl)benzenesulfonamide (Compound 101);

N-(3-Fluorophenyl)-3-(3H-imidazo[4,5-b]pyridin-6-yl)-4-methoxybenzenesulfonamide (Compound 102);

4-Methoxy-3-(4-oxo-1,4-dihydroquinolin-6-yl)-N-(m-tolyl)benzenesulfonamide (Compound 103);

N-(3-fluorophenyl)-4-methoxy-3-(4-oxo-1,4-dihydroquinolin-6-yl)benzenesulfonamide (Compound 104); and

3-(4-oxo-1,4-dihydroquinolin-6-yl)-N-(m-tolyl)benzenesulfonamide (Compound 105); is selected from the group consisting of:

The present invention relates to a pharmaceutical composition used for treating, preventing, inhibiting, or eliminating an IRP2-dependent disease or disorder, comprising a compound represented by the above chemical formula 1, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

The present invention relates to a pharmaceutical composition for the prevention or treatment of solid cancer, which is an IKZF2-dependent disease, comprising a compound represented by the above chemical formula 1, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

The solid cancer may be selected from the group consisting of non-small cell lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, carcinoid, gastrointestinal stromal tumor (GIST), prostate cancer, breast cancer, lymphoma, leukemia, melanoma, bladder carcinoma, colon cancer, cutaneous melanoma, hepatocellular carcinoma, endometrial cancer, ovarian cancer, cervical cancer, lung cancer, renal cancer, glioblastoma multiforme, glioma, thyroid cancer, parathyroid cancer, nasopharyngeal cancer, tongue cancer, pancreatic cancer, esophageal cancer, cholangiocarcinoma, gastric cancer, soft tissue sarcoma, rhabdomyosarcoma (RMS), synovial sarcoma, osteosarcoma, hepatic carcinoma, and Ewing sarcoma, and is preferably colon cancer.

The compound represented by the chemical formula 1 of the present invention, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof can enhance the cancer treatment effect by combination therapy with an anticancer agent for the treatment of solid cancer, and the anticancer agent that can be used in the combination therapy can be selected from taxane-based anticancer agents, antitumor alkylating agents, antitumor antimetabolites, antitumor antibiotics, plant-derived antitumor agents, antitumor platinum complexes, antitumor camptothecin derivatives, antitumor kinase inhibitors, antitumor antibodies, hormonal antitumor agents, antitumor viral agents, and angiogenesis inhibitors.

The above taxane anticancer agent is paclitaxel, docetaxel, cabazitaxel, larotaxel, BMS-184476, BMS-188797, BMS275183, milataxel, ortaxel, TL-310, docosahexaenoic acid-paclitaxel (DHA-paclitaxel), nab-paclitaxel, EndoTAG+paclitaxel, XRP9881, polymer-micelle paclitaxel or RPR-109881A, the above antitumor alkylating agent is nitrogen mustard N-oxide, cyclophosphamide, ifosfamide, melphalan, busulan, mitobronitol, carboquone, thiotepa, ranimustine, nimustine, temozolomide or carmustine, and the above antitumor antimetabolite is methotrexate, 6-mercaptopurine riboside, Mercaptopurine, 5-fluorouracil, tegafur, doxifluridine, camofur, cytarabine, cytarabine ocphosphate, enocitabine, S-1, gemcitabine, fludarabine or pemetrexed disodium, and the antitumor antibiotic is actinomycin D, doxorubicin, daunorubicin, neocasinostatin, bleomycin, peplomycin, mitomycin C, aclarubicin, pirarubicin, epirubicin, zinostatin stimalamer, idarubicin, sirolimus or valrubicin, and the plant-derived antitumor agent is vincristine, vinblastine, vindesine, etoposide, sobuzoxane, docetaxol, paclitaxel or vinorelbine, and the antitumor platinum complex is cisplatin, carboplatin, nedaplatin or Oxaliplatin, the antitumor camptothecin derivative is irinotecan, topotecan or camptothecin, the antitumor kinase inhibitor is gefitinib, imatinib or erlotinib, the antitumor antibody is cetuximab, bevacizumab, rituximab, bevacizumab, alemtuzumab or trastuzumab, the hormonal antitumor agent is goseserelin, leuprolide or tamoxifen, the antitumor viral agent is Imlygic, and the angiogenesis inhibitor is avastin, bevacizumab, ramucirumab, aflibercept, cetuximab, panitumumab, regorafenib, sunitinib, sorafenib, pazopanib, vandetanib, axitinib, cediranib, It may be, but is not particularly limited to, vatalanib, motesanib, lucatinib, intedanib, semaxanib, afatinib, lenvatinib, cabozantinib, or a combination thereof.

Additionally, the following terms in the present invention have the following meanings unless otherwise specified. Any undefined term has the meaning understood in the art.

The above term “halogen” refers to fluorine (F), chlorine (Cl), bromine (Br), and iodine (I).

The term “alkyl” above refers to a straight or branched chain hydrocarbon group with a single bond. Examples include methyl, ethyl, propyl, n-butyl, isobutyl, tert-butyl, and 1-methylpropyl.

The term “amino” above means -NH ₂ .

The term “alkoxy” above refers to an oxygen group bonded to a straight or branched chain saturated hydrocarbon group with a single bond. Examples include methoxy, ethoxy, propoxy, n-butoxy, tert-butoxy, and 1-methylpropoxy.

The term “cycloalkyl” above refers to a saturated hydrocarbon group with a single bond in a ring shape. Examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

The term “aryl” as used herein means an aromatic substituent having at least one ring having a shared pi electron system, including but not limited to phenyl, benzyl, and the like.

The above term “heterocycloalkyl” refers to a saturated hydrocarbon group with a single bond in a ring shape containing one or more heteroatoms such as N, O, or S, and includes, but is not limited to, aziridinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, tetrahydropyranyl, etc., depending on the number and type of heteroatoms contained in the ring and the number of carbon atoms.

The above term “heteroaryl” refers to an aromatic ring compound containing one or more heteroatoms such as N, O, or S, and includes, but is not limited to, pyrrolyl, furanyl, pyridinyl, pyrimidinyl, pyranyl, pyrazolyl, thiophenyl (or thienyl), etc., depending on the number and type of heteroatoms contained in the ring and the number of carbon atoms.

The term “haloalkyl” above means an alkyl group substituted with a halo group, where halo and alkyl are as disclosed above.

The term “alkylamino” above means an amino group substituted with an alkyl group, wherein alkyl and amino are as disclosed above.

The term “dialkylamino” above means an amino group substituted with two alkyl groups, wherein alkyl and amino are as disclosed above.

The above term “dialkylaminoalkyl” means an alkyl group substituted with a dialkylamino group, where dialkylamino and alkyl are as disclosed above.

The above term “arylamino” means an amino group substituted with an aryl group, wherein aryl and amino are as disclosed above.

In the present invention, the colon cancer may collectively refer to rectal cancer, colon cancer, and anal cancer, but is not particularly limited thereto.

In the present invention, the pharmaceutically acceptable salt refers to a salt or complex of chemical formula 1 having desirable biological activity. Examples of such salts include, but are not limited to, acid addition salts formed with inorganic acids (e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, etc.), and acid addition salts formed with acetic acid, oxalic acid, tartaric acid, succinic acid, malic acid, fumaric acid, maleic acid, ascorbic acid, benzoic acid, tannic acid, pamoic acid, alginic acid, polyglutamic acid, naphthalene sulfonic acid, naphthalene disulfonic acid, and poly-galacturonic acid. Salts formed with the same organic acid are included. The compound may also be administered as a pharmaceutically acceptable quaternary salt known to those skilled in the art, particularly chloride, bromide, iodide, -O-alkyl, toluenesulfonate, methylsulfonate, sulfonate, phosphate, or carboxylate (e.g., benzoate, succinate, acetate, glycolate, maleate, malate, fumarate, citrate, tartrate, ascorbate, cinnamoate, mandeloate, and diphenylacetate). The compound of formula 1 of the present invention may include not only pharmaceutically acceptable salts, but also all salts, hydrates, solvates, and prodrugs that can be prepared by conventional methods.

The acid addition salt according to the present invention can be prepared by a conventional method, for example, by dissolving the derivative of chemical formula 1 in an organic solvent such as methanol, ethanol, acetone, dichloromethane, acetonitrile, etc., adding an organic acid or inorganic acid, filtering and drying the resulting precipitate, or by distilling the solvent and an excess acid under reduced pressure, drying, and crystallizing in the presence of an organic solvent.

Additionally, pharmaceutically acceptable metal salts can be prepared using bases. Alkali metal or alkaline earth metal salts can be obtained, for example, by dissolving a compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering out the undissolved compound salt, and evaporating and drying the filtrate. Among the metal salts, sodium, potassium, or calcium salts are pharmaceutically suitable. Furthermore, the corresponding salts can be obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (e.g., silver nitrate).

Furthermore, the compounds of the present invention may contain one or more asymmetric carbon atoms and may exist in racemic and optically active forms. All such compounds and diastereomers are included within the scope of the present invention.

The pharmaceutical composition according to the present invention can be formulated into a suitable form together with a pharmaceutically acceptable carrier commonly used. "Pharmaceutically acceptable" refers to a composition that is physiologically tolerable and does not typically cause allergic reactions or similar reactions such as gastrointestinal disorders or dizziness when administered to humans. In addition, the composition can be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, and sterile injectable solutions, each according to a conventional method.

Carriers, excipients and diluents that may be included in the above composition include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl parahydroxybenzoate, propyl parahydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, the composition is prepared using diluents or excipients such as commonly used fillers, stabilizers, binders, disintegrants and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations are prepared by mixing the compound of the present invention with at least one excipient, such as starch, microcrystalline cellulose, sucrose or lactose, low-substituted hydroxypropyl cellulose, hypromellose, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, oral solutions, emulsions, syrups, etc., and in addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, flavoring agents, preservatives, etc. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspending agents may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases may include witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerol, gelatin, etc. In order to formulate a dosage form for parenteral administration, the benzenesulfonamide-substituted heterobicyclic derivative compound of the above chemical formula 1 or a pharmaceutically acceptable salt thereof is sterilized and/or mixed in water with auxiliary agents such as preservatives, stabilizers, wetting agents or emulsifying promoters, salts for osmotic pressure control and/or buffers, and other therapeutically useful substances to prepare a solution or suspension, which may be prepared in an ampoule or vial unit dosage form.

The above pharmaceutical composition provides a pharmaceutical composition comprising a benzenesulfonamide substituted heterobicyclic derivative compound of the above chemical formula 1 and an excipient. The compound may be added in an amount of preferably 0.001 wt% to 50 wt%, more preferably 0.001 wt% to 40 wt%, and most preferably 0.001 wt% to 30 wt% based on the total weight of the entire composition.

The pharmaceutical composition comprising the compound of chemical formula 1 disclosed in the present invention as an active ingredient can be administered to mammals such as mice, livestock, and humans by various routes. All modes of administration are conceivable, and for example, it can be administered orally, rectally, or by intravenous, intramuscular, subcutaneous, intrauterine, or intracerebrovascular injection. The dosage will vary depending on the age, sex, and body weight of the subject to be treated, the specific disease or pathological condition to be treated, the severity of the disease or pathological condition, the time of administration, the route of administration, the absorption, distribution, and excretion rates of the drug, the types of other drugs used, and the judgment of the prescriber. Determination of the dosage based on these factors is within the level of those skilled in the art, and the dosage generally ranges from 0.01 mg/kg/day to approximately 2000 mg/kg/day. A more preferred dosage is 1 mg/kg/day to 500 mg/kg/day. The dosage can be administered once a day or divided into several doses. The above dosage does not limit the scope of the present invention in any way.

The present invention relates to a pharmaceutical composition for preventing or treating colon cancer, comprising a benzenesulfonamide-substituted heterobicyclic derivative, which is an IRP2 disruptor, as an active ingredient, and can provide a benzenesulfonamide-substituted heterobicyclic derivative compound that kills colon cancer cells by inducing reprogramming of iron metabolism in cancer cells by interfering with IRP2 binding to IRE.

Figure 1 shows the results showing that IRP2 is an important factor regulating cell growth and has therapeutic relevance in colon cancer.
Figure 2 shows the results of identifying small molecule inhibitors with strong selectivity for IRP2.
Figure 3 shows the results showing changes in iron metabolism-related factors through cancer cell toxicity and IRP2 inhibition induced by compound 1 and compound 44.
Figure 4 shows the results of IRP2 ubiquitination and sequential LIP, ROS reduction, and mitochondrial OCR reduction by IRP2 inhibitor.
Figure 5 shows the results of gene profiling and GSEA analysis showing a decrease in mitochondrial OCR genes and an increase in autophagy genes by compound 1.
Figure 6 shows the results of inducing autophagic cancer cell death through activation of the AMPK-ULK1-Beclin1-LC3B pathway by compound 1 and compound 44.
Figure 7 shows the results of inhibiting tumor growth in organoid and in vivo animal models of compound 1 and compound 44.
Figure 8 shows the results of examining iron metabolism-related factors by changing the amount of intracellular iron using FAC and DFO.
Figure 9 shows the results of confirming the mRNA and protein expression levels of IRP1 and IRP2 in normal and cancerous tissues of a patient.
Figure 10 shows the results of compound hits from the pharmacophore-based screening of the Database (DB).
Figure 11 is a synthetic route scheme for compound 1 (A) and compound 44 (B).
Figure 12 shows the effects on transcriptional activity and cell cycle after treatment with compound 1 and compound 44.
Figure 13 shows the results showing mitochondrial dysfunction and autophagy-dependent cancer cell death induced by compound 1.

Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Rather, the content presented herein is intended to be thorough and complete, and to fully convey the spirit of the present invention to those skilled in the art.

<Example 1. Synthesis of benzenesulfonamide-substituted heterobicyclic derivative compounds and confirmation of physicochemical properties>

Compounds 1 to 105 of the present invention synthesized various benzenesulfonamide-substituted heterobicyclic derivatives, and their physicochemical properties are as follows.

Compound 1. 4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide

1-1. Synthesis of 3-bromo-4-methoxybenzenesulfonyl chloride (compound 1-1)

2-Bromoanisole (12.4 mL, 100.0 mmol) and DCM (200 mL) were added and stirred at 0 °C, then Chlorosulfonic acid (20 mL, 300.0 mmol) (3.0 eq.) was slowly added and stirred at room temperature for 12 h. After completion of the reaction, the reactant was slowly added to ice water, extracted with DCM, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure to obtain compound 1-1 in a yield of 99% (28.3 g).

1-2. Synthesis of 3-bromo-4-methoxy-N-(m-tolyl)benzenesulfonamide (Compound 1-2)

Compound 1-1 (3-bromo-4-methoxybenzenesulfonyl chloride; 14.4 g, 50.0 mmol), THF (150 mL), and pyridine (4.0 mL, 50.0 mmol)(1.0 eq.) were added and stirred at 0 °C, then m-toluidine (6.4 mL, 60.0 mmol)(1.5 eq.) was added and stirred at room temperature for 12 h. After completion of the reaction, the mixture was extracted with _H2O and EtOAc, and the organic layer was dried over _MgSO4 and concentrated under reduced pressure. After that, compound 1-2 was obtained in a yield of 90% (16 g) by separation by silica gel column chromatography.

1-3. Synthesis of 4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzene sulfonamide (Compound 1)

5-Bromo-1H-pyrazolo[3,4-b]pyridine (990 mg, 5 mmol), bis(pinacolato)diboron (1.52 g, 6 mmol)(1.2 eq.), [1,1′-Bis(diphenylphosphino)ferrocene]dichloropalladium(II) dichloromethane complex (408 mg, 0.5 mmol)(10 mol%), potassium acetate (1.47 g, 15 mmol)(3.0 eq.), and 1,4-dioxane (10 mL) were added and reacted using a microwave reactor at 120 °C for 1 hour. After cooling to room temperature, compound 1-2 (3-bromo-4-methoxy-N-(m-tolyl)benzenesulfonamide; 1.78 g, 5 mmol)(1.0 eq.) and sodium carbonate aqueous solution (2 M, 5 mL)(2.0 eq.) were added and reacted at 120 ℃ for 1 hour. After completion of the reaction, the solvent was removed by celite filtration, extracted with water and EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. After that, compound 1 was obtained in a yield of 25% (495 mg) by separation by silica gel column chromatography.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.8 (s, 1H), 10.1 (s, 1H), 8.51 (d, J = 2.1 Hz, 1H), 8.22 (d, J = 2.1 Hz, 1H), 8.21 (d, J = 1.1 Hz, 1H), 7.77 (dd, J = 8.7, 2.4 Hz, 1H), 7.71 (d, J = 2.4 Hz, 1H), 7.28 (d, J = 8.8 Hz, 1H), 7.13 (t, J = 7.7 Hz, 1H), 7.00 - 6.90 (m, 2H), 6.91 - 6.82 (m, 1H), 3.84 (s, 3H), 2.21 (s, 3H)

Compound 2. 4-Methoxy-N-phenyl-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide

Compound 2 (yield 45%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.76 (brs, 1H), 10.12 (brs, 1H), 8.51 (d, J = 2.1 Hz, 1H), 8.25 - 8.17 (m, 2H), 7.79 - 7.68 (m, 2H), 7.32 - 7.21 (m, 3H), 7.16 - 7.10 (m, 2H), 7.04 (t, J = 7.2 Hz, 1H), 3.83 (s, 3H).

Compound 3. 4-Methoxy-N-(3-methoxyphenyl)-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide

Compound 3 (yield 52%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.77 (brs, 1H), 10.16 (brs, 1H), 8.52 (d, J = 2.0 Hz, 1H), 8.26 - 8.18 (m, 2H), 7.78 (dd, J = 8.7, 2.4 Hz, 1H), 7.73 (d, J = 2.4 Hz, 1H), 7.29 (d, J = 8.8 Hz, 1H), 7.15 (t, J = 8.4 Hz, 1H), 6.75 - 6.69 (m, 2H), 6.66 - 6.59 (m, 1H), 3.84 (s, 3H), 3.66 (s, 3H).

Compound 4. 3-((4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)phenyl)sulfonamido)benzamide

Compound 4 (yield 20%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.76 (brs, 1H), 10.28 (brs, 1H), 8.55 (d, J = 2.1 Hz, 1H), 8.21 (d, J = 2.1 Hz, 2H), 7.96 (brs, 1H), 7.77 (dd, J = 8.6, 2.5 Hz, 1H), 7.72 (d, J = 2.4 Hz, 1H), 7.68 (d, J = 2.1 Hz, 1H), 7.54 (d, J = 7.3 Hz, 1H), 7.41 (brs, 1H), 7.36 - 7.24 (m, 3H), 3.83 (s, 3H).

Compound 5. 4-Methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(3-(trifluoromethyl)phenyl)benzenesulfonamide

Compound 5 (yield 63%) was synthesized using the same method as the synthesis method for compound 1 above.

¹ H NMR (300 MHz, DMSO-d ₆ ) δ 13.77 (brs, 1H), 10.60 (brs, 1H), 8.51 (s, 1H), 8.26 - 8.16 (m, 2H), 7.85 - 7.70 (m, 2H), 7.55 - 7.37 (m, 4H), 7.31 (d, J = 8.9 Hz, 1H), 3.84 (s, 3H).

Compound 6. N-(3-cyanophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide

Compound 6 (yield 34%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.77 (brs, 1H), 10.65 (brs, 1H), 8.55 (d, J = 2.1 Hz, 1H), 8.26 (d, J = 2.1 Hz, 1H), 8.20 (d, J = 1.4 Hz, 1H), 7.85 - 7.75 (m, 2H), 7.56 - 7.42 (m, 4H), 7.30 (d, J = 8.8 Hz, 1H), 3.84 (s, 3H).

Compound 7. N-(3-fluorophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide

Compound 7 (yield 58%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.77 (brs, 1H), 10.47 (brs, 1H), 8.54 (d, J = 2.1 Hz, 1H), 8.24 (dd, J = 15.3, 1.7 Hz, 2H), 7.90 - 7.69 (m, 2H), 7.39 - 7.20 (m, 2H), 7.06 - 6.80 (m, 3H), 3.85 (s, 3H).

Compound 8. N-(4-fluorophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide

Compound 8 (yield 34%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.76 (brs, 1H), 10.09 (brs, 1H), 8.53 (d, J = 2.1 Hz, 1H), 8.25 (d, J = 2.2 Hz, 1H), 8.20 (d, J = 1.3 Hz, 1H), 7.70 (d, J = 7.4 Hz, 2H), 7.31 - 7.24 (m, 1H), 7.16 - 7.07 (m, 4H), 3.84 (s, 3H).

Compound 9. N-(2-fluorophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide

Compound 9 (yield 47%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.74 (brs, 1H), 10.05 (brs, 1H), 8.53 (d, J = 2.1 Hz, 1H), 8.24 (d, J = 2.1 Hz, 1H), 8.20 (d, J = 1.2 Hz, 1H), 7.77 - 7.67 (m, 2H), 7.34 - 7.24 (m, 2H), 7.20 - 7.06 (m, 3H), 3.85 (s, 3H).

Compound 10. N-(3-chlorophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide

Compound 10 (yield 68%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.77 (brs, 1H), 10.45 (brs, 1H), 8.54 (s, 1H), 8.28 - 8.17 (m, 2H), 7.85 - 7.69 (m, 2H), 7.29 (t, J = 9.3 Hz, 2H), 7.20 - 7.04 (m, 3H), 3.84 (s, 3H).

Compound 11. N-(4-chlorophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide

Compound 11 (yield 34%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.76 (brs, 1H), 10.09 (brs, 1H), 8.53 (d, J = 2.1 Hz, 1H), 8.25 (d, J = 2.2 Hz, 1H), 8.20 (d, J = 1.3 Hz, 1H), 7.70 (d, J = 7.4 Hz, 2H), 7.31 - 7.24 (m, 1H), 7.16 - 7.07 (m, 4H), 3.84 (s, 3H).

Compound 12. N-(2-chlorophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide

Compound 12 (yield 47%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.74 (brs, 1H), 10.05 (brs, 1H), 8.53 (d, J = 2.1 Hz, 1H), 8.24 (d, J = 2.1 Hz, 1H), 8.20 (d, J = 1.2 Hz, 1H), 7.77 - 7.67 (m, 2H), 7.34 - 7.24 (m, 2H), 7.20 - 7.06 (m, 3H), 3.85 (s, 3H).

Compound 13. N-(2-cyclopropylphenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide

13-1. Synthesis of 2-cyclopropylaniline (compound 13-1)

2-Bromoaniline (1.72 g, 10.0 mmol), cyclopropylboronic acid (1.12 g, 13.0 mmol)(1.3 eq.), palladium(II) acetate (220 mg, 1.0 mmol)(0.1 eq.), triphenylphosphine (260 mg, 1.0 mmol)(0.1 eq.), potassium carbonate (4.84 g, 35.0 mmol)(3.5 eq.), and toluene/H ₂ O (50/5 mL) were added and stirred at 110°C for 24 h. After completion of the reaction, the solvent was removed by celite filtration, extracted with water and EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. Compound 13-1 was obtained in a yield of 30% (405 mg) through separation by silica gel column chromatography.

13-2. Synthesis of N-(2-cyclopropylphenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 13)

Compound 13 (yield 3%) was synthesized using the same method as synthetic methods 1-2 and 1-3 of the above compound 1.

^1H NMR (400 MHz, Acetone-d ₆ ) δ 12.81 (s, 1H), 8.55 (d, J = 2.1 Hz, 1H), 8.26 (s, 1H), 8.20 (d, J = 2.1 Hz, 1H), 8.16 (s, 1H), 7.79 (dd, J = 8.7, 2.4 Hz, 1H), 7.69 (d, J = 2.4 Hz, 1H), 7.43 (dd, J = 7.6, 1.8 Hz, 1H), 7.29 (d, J = 8.7 Hz, 1H), 7.22 - 7.12 (m, 2H), 6.95 (dd, J = 7.4, 1.9 Hz, 1H), 3.95 (s, 3H), 1.97 - 1.89 (m, 2H), 0.85 - 0.78 (m, 2H), 0.48 - 0.41 (m, 2H).

Compound 14. N-(4-hydroxyphenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide

Compound 14 (yield 10%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.76 (s, 1H), 9.59 (s, 1H), 9.35 (s, 1H), 8.54 (d, J = 2.0 Hz, 1H), 8.20 (d, J = 2.2 Hz, 2H), 7.66 (dd, J = 8.7, 2.4 Hz, 1H), 7.60 (d, J = 2.4 Hz, 1H), 7.27 (d, J = 8.7 Hz, 1H), 6.94 6.86 (m, 2H), 6.65 (d, J = 8.7 Hz, 2H), 3.85 (s, 3H).

Compound 15. N-(3-(2-hydroxyethoxy)phenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide

15-1. Synthesis of 2-(3-aminophenoxy)ethan-1-ol (Compound 15-1)

m-Nitrophenol (1.39 g, 10.0 mmol), sodium hydroxide (8 g, 200 mmol)(20.0 eq.), 2-chloroethanol (13.4 mL, 200 mmol)(20.0 eq.) and water (50 mL) were added and stirred at 80 ℃ for 3 h. After completion of the reaction, the mixture was extracted with water and EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. To this was added MeOH (20 mL) and Palladium on carbon (360 mg, 20 wt%), a hydrogen balloon was connected, and the mixture was stirred at room temperature for 4 h. After completion of the reaction, the filtrate was filtered through celite, and the filtrate was concentrated under reduced pressure to obtain compound 15-1 in a yield of 68% (1.05 g, 2 steps).

15-2. Synthesis of N-(3-(2-hydroxyethoxy)phenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 15)

Compound 15 (yield 3%) was synthesized using the same method as synthetic methods 1-2 and 1-3 of the above compound 1.

^1H NMR (300 MHz, Acetone-d ₆ ) δ 12.83 (s, 1H), 8.98 - 8.93 (m, 1H), 8.60 (s, 1H), 8.26 (d, J = 1.7 Hz, 1H), 8.19 (s, 1H), 7.86 (dd, J = 8.7, 2.4 Hz, 1H), 7.81 (d, J = 2.4 Hz, 1H), 7.28 (d, J = 8.7 Hz, 1H), 7.19 (t, J = 8.2 Hz, 1H), 6.93 (t, J = 2.2 Hz, 1H), 6.85 (ddd, J = 8.0, 2.1, 0.9 Hz, 1H), 6.70 (ddd, J = 8.3, 2.5, 0.9 Hz, 1H), 3.93 (d, J = 0.9 Hz, 3H), 3.85 (q, J = 5.0 Hz, 2H), 2.88 (d, J = 10.1 Hz, 2H).

Compound 16. N-(4-(2-hydroxyethoxy)phenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide

After the same process as in Synthesis Method 15-1 of the above compound 15, compound 16 (yield 5%) was synthesized using the same method as in Synthesis Methods 1-2 and 1-3 of the above compound 1.

^1H NMR (300 MHz, Acetone-d ₆ ) δ 12.84 (s, 1H), 8.61 (s, 1H), 8.55 (d, J = 2.1 Hz, 1H), 8.21 (d, J = 2.1 Hz, 1H), 8.18 (s, 1H), 7.75 (dd, J = 8.7, 2.4 Hz, 1H), 7.65 (d, J = 2.4 Hz, 1H), 7.26 (d, J = 8.7 Hz, 1H), 7.20 - 7.11 (m, 2H), 6.93 - 6.86 (m, 2H), 4.11 - 4.05 (m, 2H), 3.93 (s, 3H), 3.90 - 3.82 (m, 2H).

Compound 17. N-(2-(2-hydroxyethoxy)phenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide

After the same process as in Synthesis Method 15-1 of the above compound 15, compound 17 (yield 1%) was synthesized using the same method as in Synthesis Methods 1-2 and 1-3 of the above compound 1.

^1H NMR (300 MHz, Acetone-d ₆ ) δ 12.79 (s, 1H), 8.56 (d, J = 2.0 Hz, 1H), 8.35 (s, 1H), 8.22 (d, J = 2.0 Hz, 1H), 8.17 (s, 1H), 7.79 (dd, J = 8.7, 2.4 Hz, 1H), 7.70 (d, J = 2.4 Hz, 1H), 7.63 - 7.57 (m, 1H), 7.25 (d, J = 8.7 Hz, 1H), 7.18 - 7.09 (m, 1H), 6.98 (td, J = 7.8, 1.1 Hz, 2H), 3.92 (s, 3H), 3.91 - 3.87 (m, 2H), 3.70 (q, J = 5.2 Hz, 2H).

Compound 18. 4-Methoxy-N-(4-(4-methylpiperazin-1-yl)phenyl)-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide

Compound 18 (yield 12%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.77 (brs, 1H), 9.64 (brs, 1H), 8.50 (d, J = 2.0 Hz, 1H), 8.23 - 8.15 (m, 2H), 7.68 (dd, J = 8.7, 2.4 Hz, 1H), 7.57 (d, J = 2.3 Hz, 1H), 7.27 (d, J = 8.9 Hz, 1H), 6.93 (d, J = 9.1 Hz, 2H), 6.82 (d, J = 9.2 Hz, 2H), 3.84 (s, 3H), 3.03 (m, 4H), 2.39 (m, 4H), 2.19 (s, 3H).

Compound 19. 4-Methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(pyridin-3-yl)benzenesulfonamide

Compound 19 (yield 38%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.77 (brs, 1H), 10.43 (brs, 1H), 8.54 (d, J = 2.1 Hz, 1H), 8.31 (d, J = 2.0 Hz, 1H), 8.27 (dd, J = 4.9, 1.7 Hz, 2H), 8.21 (d, J = 1.4 Hz, 1H), 7.80 - 7.72 (m, 2H), 7.56 (ddd, J = 8.3, 2.8, 1.6 Hz, 1H), 7.36 - 7.26 (m, 2H), 3.84 (s, 3H).

Compound 20. N-(3-fluorophenyl)-3-(4-oxo-1,4-dihydroquinolin-6-yl)benzenesulfonamide

Compound 20 (yield 17%) was synthesized using the same method as the step B synthesis of compound 26 below.

^1H NMR (400 MHz, DMSO _-d6 ) δ 11.93 (s, 1H), 10.56 (s, 1H), 8.36 (d, J = 2.3 Hz, 1H), 8.13 (d, J = 2.0 Hz, 1H), 8.00-7.91 (m, 3H), 7.75 (dd, J = 7.8, 1.7 Hz, 1H), 7.71-7.62 (m, 2H), 7.29-7.21 (m, 1H), 6.98-6.89 (m, 2H), 6.81 (td, J = 8.6, 2.6 Hz, 1H), 6.10 (d, J = 7.4 Hz, 1H).

Compound 21. 5-(5-((1H-pyrazol-1-yl)sulfonyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridine

Compound 21 (yield 12%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.77 (brs, 1H), 8.59 (d, J = 2.1 Hz, 1H), 8.50 (d, J = 2.3 Hz, 1H), 8.32 (d, J = 2.1 Hz, 1H), 8.20 (s, 1H), 8.03 (dd, J = 8.8, 2.5 Hz, 1H), 7.92 (d, J = 2.5 Hz, 1H), 7.89 (d, J = 1.7 Hz, 1H), 7.41 (d, J = 9.0 Hz, 1H), 6.59 (dd, J = 2.8, 1.6 Hz, 1H), 3.89 (s, 3H).

Compound 22. 5-(2-methoxy-5-((4-methylpiperazin-1-yl)sulfonyl)phenyl)-1H-pyrazolo[3,4-b]pyridine

Compound 22 (yield 23%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.75 (brs, 1H), 8.64 (d, J = 2.1 Hz, 1H), 8.36 (d, J = 2.1 Hz, 1H), 8.20 (s, 1H), 7.77 (dd, J = 8.7, 2.4 Hz, 1H), 7.65 (d, J = 2.4 Hz, 1H), 7.40 (d, J = 8.8 Hz, 1H), 3.90 (s, 3H), 2.92 (m, 4H), 2.38 (m, 4H), 2.14 (s, 3H).

Compound 23. 4-((4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)phenyl)sulfonyl)morpholine

Compound 23 (yield 21%) was synthesized using the same method as the synthesis of compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.76 (brs, 1H), 8.66 (d, J = 2.1 Hz, 1H), 8.37 (d, J = 2.1 Hz, 1H), 8.20 (s, 1H), 7.78 (dd, J = 8.7, 2.4 Hz, 1H), 7.66 (d, J = 2.4 Hz, 1H), 7.42 (d, J = 8.8 Hz, 1H), 3.91 (s, 3H), 3.70 - 3.59 (m, 4H), 2.90 (m, 4H).

Compound 24. N-(3,5-difluorophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide

5-Bromo-1H-pyrazolo[3,4-b]pyridine (198 mg, 1.0 mmol), Bis(pinacolato)diboron (305 mg, 1.2 mmol), [1,1′-Bis(diphenylphosphino)ferrocene]dichloropalladium(II) dichloromethane complex (82 mg, 0.1 mmol)(10 mol%), potassium acetate (294 mg, 3.0 mmol), and 1,4-Dioxane (5 mL) were added, degassing was performed with N ₂ for 10 min, and stirring was performed at 100 °C for 12 h. After the reaction was completed, the solvent was removed by celite filtration, and 3-bromo-N-(3,5-difluorophenyl)-4-methoxybenzenesulfonamide (378 mg, 1.0 mmol), Bis(diphenylphosphino)ferrocene]dichloropalladium (73 mg, 0.1 mmol), sodium carbonate aqueous solution (2.0 M, 1 mL), and 1,4-Dioxane (5 mL) were added and stirred at 100 ℃ for 12 hours. After the reaction was completed, the solvent was removed by celite filtration, and the mixture was extracted with water and EtOAc. The organic layer was dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain compound 24 in a yield of 48% (200 mg).

^1H NMR (300 MHz, DMSO _-d6 ) δ 13.77 (s, 1H), 10.77 (s, 1H), 8.56 (d, J = 2.1 Hz, 1H), 8.28 (d, J = 2.1 Hz, 1H), 8.21 (d, J = 1.3 Hz, 1H), 7.86 (dd, J = 8.7, 2.5 Hz, 1H), 7.80 (d, J = 2.4 Hz, 1H), 7.33 (d, J = 8.8 Hz, 1H), 6.94-6.86 (m, 1H), 6.85-6.75 (m, 2H), 3.86 (s, 3H).

Compound 25. N-(3-chloro-5-fluorophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide

Compound 25 (yield 42%) was synthesized using the same method as the synthesis of compound 24 above.

^1H NMR (300 MHz, DMSO _-d6 ) δ 13.77 (s, 1H), 10.76 (s, 1H), 8.56 (d, J = 2.1 Hz, 1H), 8.28 (d, J = 2.1 Hz, 1H), 8.21 (d, J = 1.3 Hz, 1H), 7.85 (dd, J = 8.7, 2.5 Hz, 1H), 7.79 (d, J = 2.4 Hz, 1H), 7.34 (d, J = 8.8 Hz, 1H), 7.12-7.08 (m, 1H), 7.01-6.99 (m, 1H), 6.97-6.92 (m, 1H), 3.86 (s, 3H).

Compound 26. 4-Methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzamide

Step A) Preparation of 3-bromo-4-methoxy-N-(m-tolyl)benzamide

3-Bromo-4-methoxybenzoic acid (500 mg, 2.16 mmol), m-toluidine (280 μL, 2.59 mmol), HATU (1.23 g, 3.25 mmol), triethylamine (280 μL, 4.33 mmol), and DMF (10 mL) were stirred at 30 °C for 16 h. After the reaction was completed, the mixture was extracted with water and DCM, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. The compound was then purified by silica gel column chromatography to obtain the compound in a yield of 75% (520 mg).

Step B) Preparation of 4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzamide (compound 26)

5-Bromo-1H-pyrazolo[3,4-b]pyridine (99 mg, 0.5 mmol), Bis(pinacolato)diboron (152 mg, 0.6 mmol), [1,1′-Bis(diphenylphosphino)ferrocene]dichloropalladium(II) dichloromethane complex (41 mg, 0.05 mmol), potassium acetate (147 mg, 1.5 mmol), and 1,4-Dioxane (5 mL) were added, degassed with _N2 for 10 min, and stirred at 105 ℃ for 12 h. After cooling to room temperature, 3-bromo-4-methoxy-N-(m-tolyl)benzamide (160 mg, 0.5 mmol) and aqueous sodium carbonate solution (2.0 M, 0.5 mL) were added, and stirred at 105 ℃ for 12 h. After the reaction was completed, the solvent was removed by celite filtration, extracted with water and EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. After separation by silica gel column chromatography, compound 26 was obtained with a yield of 52% (94 mg).

^1H NMR (300 MHz, DMSO _-d6 ) δ 13.73 (s, 1H), 10.07 (s, 1H), 8.71 (d, J = 2.1 Hz, 1H), 8.38 (d, J = 2.1 Hz, 1H), 8.21 (d, J = 1.3 Hz, 1H), 8.10-8.00 (m, 2H), 7.62-7.52 (m, 2H), 7.30 (d, J = 8.5 Hz, 1H), 7.22 (t, J = 7.7 Hz, 1H), 6.91 (d, J = 7.5 Hz, 1H), 3.89 (s, 3H), 2.31 (s, 3H).

Compound 27. N-(3-fluorophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzamide

Compound 27 (yield 40%) was synthesized using the same method as the step B synthesis of compound 26 above.

^1H NMR (300 MHz, DMSO _-d6 ) δ 13.73 (s, 1H), 10.32 (s, 1H), 8.70 (d, J = 2.0 Hz, 1H), 8.38 (d, J = 2.0 Hz, 1H), 8.21 (s, 1H), 8.05 (d, J = 7.5 Hz, 2H), 7.76 (dt, J = 12.0, 2.2 Hz, 1H), 7.56 (d, J = 8.2 Hz, 1H), 7.44-7.27 (m, 2H), 6.92 (dt, J = 9.6, 4.7 Hz, 1H), 3.89 (s, 3H).

Compound 28. N-phenyl-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide

Compound 28 (yield 44%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.82 (brs, 1H), 10.31 (brs, 1H), 8.77 (d, J = 2.2 Hz, 1H), 8.47 (d, J = 2.3 Hz, 1H), 8.25 (s, 1Hz), 8.07 (t, J = 1.9 Hz, 1H), 8.00 (dt, J = 7.6, 1.6 Hz, 1H), 7.84 - 7.57 (m, 3H), 7.25 (dd, J = 8.5, 7.2 Hz, 2H), 7.17 - 7.09 (m, 2H), 7.04 (t, J = 7.3 Hz, 1H).

Compound 29. 3-(1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide

Compound 29 (yield 41%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.82 (s, 1H), 10.23 (s, 1H), 8.76 (d, J = 2.2 Hz, 1H), 8.46 (d, J = 2.2 Hz, 1H), 8.24 (d, J = 1.0 Hz, 1H), 8.06 (t, J = 1.8 Hz, 1H), 8.00 (dt, J = 7.6, 1.5 Hz, 1H), 7.76 (dt, J = 7.9, 1.5 Hz, 1H), 7.67 (t, J = 7.7 Hz, 1H), 7.17 - 7.06 (m, 1H), 6.95 (d, J = 7.8 Hz, 2H), 6.86 (d, J = 7.5 Hz, 1H), 2.20 (s, 3H).

Compound 30. N-(3-fluorophenyl)-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide

Compound 30 (yield 38%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.81 (s, 1H), 10.60 (s, 1H), 8.79 (d, J = 2.2 Hz, 1H), 8.49 (d, J = 2.2 Hz, 1H), 8.24 (d, J = 1.3 Hz, 1H), 8.12 (d, J = 1.9 Hz, 1H), 8.03 (dt, J = 7.9, 1.4 Hz, 1H), 7.79 (dt, J = 7.9, 1.4 Hz, 1H), 7.69 (t, J = 7.8 Hz, 1H), 7.29 (td, J = 8.4, 6.7 Hz, 1H), 7.02 - 6.90 (m, 2H), 6.86 (td, J = 8.4, 2.6 Hz, 1H).

Compound 31. N-(3-chlorophenyl)-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide

Compound 31 (yield 46%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.81 (s, 1H), 10.59 (s, 1H), 8.79 (d, J = 2.2 Hz, 1H), 8.49 (d, J = 2.2 Hz, 1H), 8.24 (d, J = 1.3 Hz, 1H), 8.10 (t, J = 1.8 Hz, 1H), 8.03 (dt, J = 7.7, 1.5 Hz, 1H), 7.78 (dt, J = 7.9, 1.4 Hz, 1H), 7.70 (t, J = 7.7 Hz, 1H), 7.28 (t, J = 8.0 Hz, 1H), 7.17 (t, J = 2.0 Hz, 1H), 7.11 (dddd, J = 8.9, 7.9, 2.1, 1.0 Hz, 2H).

Compound 32. 3-(1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(o-tolyl)benzenesulfonamide

Compound 32 (yield 12%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.80 (s, 1H), 9.61 (s, 1H), 8.73 (d, J = 2.2 Hz, 1H), 8.42 (d, J = 2.2 Hz, 1H), 8.26 - 8.22 (m, 1H), 8.04 (dt, J = 5.5, 2.2 Hz, 1H), 7.93 - 7.90 (m, 1H), 7.70 - 7.65 (m, 2H), 7.13 (tt, J = 9.3, 3.7 Hz, 3H), 7.03 - 6.98 (m, 1H), 2.01 (s, 3H).

Compound 33. 4-Methoxy-3-(3-methyl-1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide

Compound 33 (yield 58%) was synthesized using the same method as the synthesis of compound 24 above.

^1H NMR (300 MHz, DMSO _-d6 ) δ 13.31 (s, 1H), 10.04 (s, 1H), 8.48 (d, J = 2.1 Hz, 1H), 8.11 (d, J = 2.1 Hz, 1H), 7.77 (dd, J = 8.7, 2.4 Hz, 1H), 7.68 (d, J = 2.4 Hz, 1H), 7.28 (d, J = 8.8 Hz, 1H), 7.13 (t, J = 7.7 Hz, 1H), 6.98-6.91 (m, 2H), 6.87 (d, J = 7.5 Hz, 1H), 3.84 (s, 3H), 2.52 (s, 3H), 2.22 (s, 3H).

Compound 34. 4-Methoxy-3-(1-phenyl-1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide

Compound 33 (123 mg, 0.3 mmol), phenylboronic acid (55 mg, 0.45 mmol), Cu(OAc) ₂ (16 mg, 0.09 mmol), pyridine (70 μL, 0.9 mmol), and DMF (3 mL) were added and stirred at 90 °C for 12 h. After completion of the reaction, compound 34 was obtained in 58% yield by separation using silica gel column chromatography.

^1H NMR (300 MHz, Chloroform- d ) δ 8.65 (d, J = 2.1 Hz, 1H), 8.25-8.22 (m, 2H), 7.94 (d, J = 2.1 Hz, 1H), 7.81 (dd, J = 8.7, 2.4 Hz, 1H), 7.73 (d, J = 2.4 Hz, 1H), 7.70 (s, 1H), 7.50 (t, J = 7.9 Hz, 2H), 7.30-7.26 (m, 1H), 7.13 (t, J = 7.7 Hz, 1H), 7.01-6.99 (m, 2H), 6.94 (t, J = 6.8 Hz, 2H), 3.85 (s, 3H), 2.65 (s, 3H), 2.28 (s, 3H).

Compound 35. N-(5-(2-methoxy-5-(N-(m-tolyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide

35-1. Synthesis of 5-bromo-1H-pyrazolo[3,4-b]pyridin-3-amine (Compound 35-1)

5-Bromo-2-chloropyridine-3-carbonitrile (10 g, 46.0 mmol), EtOH (150 mL), and hydrazine hydrate (5.6 mL, 115 mmol)(2.5 eq.) were added and stirred at 85°C for 12 h. After completion of the reaction, the solvent was removed, water was added, and the resulting solid was filtered to obtain compound 35-1 in a yield of 95% (9.3 g).

35-2. Synthesis of N-(5-bromo-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 35-2)

Compound 35-1 (5-bromo-1H-pyrazolo[3,4-b]pyridin-3-amine; 1.2 g, 5.63 mmol) and pyridine (10 mL) were stirred at 0 °C, and acetyl chloride (440 μL, 6.19 mmol) (1.1 eq.) was slowly added. The mixture was stirred at room temperature for 12 h. After completion of the reaction, the mixture was extracted with water and EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain compound 35-2 in a yield of 99% (1.43 g).

35-3. Synthesis of N-(5-(2-methoxy-5-(N-(m-tolyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 35)

Compound 35 (yield 32%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.32 (s, 1H), 10.75 (s, 1H), 10.11 (s, 1H), 8.50 (d, J = 2.1 Hz, 1H), 8.35 (d, J = 2.2 Hz, 1H), 7.76 (dd, J = 8.7, 2.4 Hz, 1H), 7.68 (d, J = 2.4 Hz, 1H), 7.28 (d, J = 8.8 Hz, 1H), 7.12 (t, J = 7.7 Hz, 1H), 6.94 (d, J = 8.8 Hz, 2H), 6.85 (d, J = 7.5 Hz, 1H), 3.83 (s, 3H), 2.21 (s, 3H), 2.11 (d, J = 13.5 Hz, 3H).

Compound 36. N-(5-(4-fluoro-3-(N-(m-tolyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide

Compound 36 (yield 6%) was synthesized using the same method as the step B synthesis method for compound 26 above.

^1H NMR (300 MHz, DMSO _-d6 ) δ 13.36 (s, 1H), 10.77 (s, 1H), 10.64 (s, 1H), 8.73 (d, J = 2.2 Hz, 1H), 8.55 (d, J = 2.3 Hz, 1H), 8.01 (d, J = 5.9 Hz, 2H), 7.55 (t, J = 9.4 Hz, 1H), 7.12 (t, J = 7.7 Hz, 1H), 7.01-6.92 (m, 2H), 6.84 (d, J = 7.6 Hz, 1H), 2.20 (s, 3H), 2.14 (s, 3H).

Compound 37. N-(5-(2-fluoro-5-(N-(m-tolyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide

Compound 37 (yield 1%) was synthesized using the same method as the step B synthesis of compound 26 above.

^1H NMR (300 MHz, DMSO _-d6 ) δ 13.43 (s, 1H), 10.81 (s, 1H), 10.26 (s, 1H), 8.57 (s, 1H), 8.52 (s, 1H), 7.92 (d, J = 6.8 Hz, 1H), 7.84-7.74 (m, 1H), 7.56 (t, J = 9.4 Hz, 1H), 7.14 (t, J = 7.7 Hz, 1H), 6.99-6.86 (m, 3H), 2.22 (s, 3H), 2.14 (s, 3H).

Compound 38. N-(5-(3-fluoro-5-(N-(m-tolyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide

Compound 38 (yield 3%) was synthesized using the same method as step B of the above compound 26.

^1H NMR (300 MHz, DMSO _-d6 ) δ 13.41 (s, 1H), 10.79 (s, 1H), 10.37 (s, 1H), 8.77 (d, J = 2.3 Hz, 1H), 8.61 (d, J = 2.3 Hz, 1H), 7.91 (d, J = 11.1 Hz, 1H), 7.86 (s, 1H), 7.52 (dd, J = 8.0, 2.3 Hz, 1H), 7.14 (t, J = 7.7 Hz, 1H), 6.99 - 6.92 (m, 2H), 6.88 (d, J = 7.6 Hz, 1H), 2.21 (s, 3H), 2.15 (s, 3H).

Compound 39. N-(5-(2,4-difluoro-5-(N-(m-tolyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide

Compound 39 (yield 17%) was synthesized using the same method as the step B synthesis of compound 26 above.

^1H NMR (300 MHz, DMSO _-d6 ) δ 13.42 (s, 1H), 10.80 (s, 1H), 10.67 (s, 1H), 8.58 (d, J = 2.1 Hz, 1H), 8.50 (s, 1H), 7.96 (t, J = 8.1 Hz, 1H), 7.71 (t, J = 10.2 Hz, 1H), 7.14 (t, J = 7.7 Hz, 1H), 7.01-6.92 (m, 2H), 6.87 (d, J = 7.5 Hz, 1H), 2.22 (s, 3H), 2.13 (s, 3H).

Compound 40. N-(5-(2-fluoro-3-(N-(m-tolyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide

Compound 40 (yield 11%) was synthesized using the same method as the step B synthesis of compound 26 above.

^1H NMR (300 MHz, DMSO- d ₆ ) δ 13.43 (s, 1H), 10.79 (s, 1H), 10.58 (s, 1H), 8.57 (d, J = 1.8 Hz, 1H), 8.52 (s, 1H), 7.90 - 7.79 (m, 2H), 7.46 (t, J = 7.8 Hz, 1H), 7.11 (dd, J = 9.0, 7.4 Hz, 1H), 6.97 - 6.90 (m, 2H), 6.85 (d, J = 7.5 Hz, 1H), 2.19 (s, 3H), 2.12 (s, 3H).

Compound 41. N-(5-(2-methoxy-5-(N-(p-tolyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide

Compound 41 (yield 13%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 10.74 (s, 1H), 8.48-8.45 (m, 1H), 8.37-8.35 (m, 1H), 7.74-7.69 (m, 2H), 7.64 (d, J = 2.4 Hz, 1H), 7.25 (d, J = 8.8 Hz, 1H), 7.08-6.96 (m, 4H), 3.82 (s, 3H), 2.13 (s, 3H), 2.08 (s, 3H).

Compound 42. N-(5-(2-methoxy-5-(N-(o-tolyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide

Compound 42 (yield 12%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 10.72 (s, 1H), 9.47 (s, 1H), 8.48 (d, J = 2.1 Hz, 1H), 8.33 (d, J = 2.1 Hz, 1H), 7.67 (dd, J = 8.7, 2.4 Hz, 1H), 7.53 (d, J = 2.4 Hz, 1H), 7.30 (d, J = 8.8 Hz, 1H), 7.20 - 7.07 (m, 4H), 7.04 - 6.99 (m, 1H), 3.87 (s, 3H), 2.14 (s, 3H), 2.06 (s, 3H).

Compound 43. N-(5-(2-methoxy-5-(N-phenylsulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide

Compound 43 (yield 21%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 10.73 (s, 1H), 10.16 (s, 1H), 8.51 (d, J = 2.1 Hz, 1H), 8.36 (d, J = 2.1 Hz, 1H), 7.76 (dd, J = 8.7, 2.4 Hz, 1H), 7.68 (d, J = 2.4 Hz, 1H), 7.27 (t, J = 8.5 Hz, 3H), 7.16 (d, J = 1.4 Hz, 1H), 7.13 (s, 1H), 7.05 (t, J = 7.3 Hz, 1H), 3.84 (s, 3H), 2.15 (s, 3H).

Compound 44. N-(5-(5-(N-(3-fluorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide

Compound 44 (yield 38%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.32 (s, 1H), 10.74 (s, 1H), 10.49 (s, 1H), 8.52 (d, J = 2.1 Hz, 1H), 8.36 (d, J = 2.1 Hz, 1H), 7.80 (dd, J = 8.7, 2.4 Hz, 1H), 7.72 (d, J = 2.4 Hz, 1H), 7.29 (td, J = 8.4, 6.3 Hz, 2H), 7.01 - 6.90 (m, 2H), 6.86 (ddd, J = 10.5, 8.3, 2.5 Hz, 1H), 3.84 (s, 3H), 2.13 (s, 3H).

Compound 45. N-(5-(4-fluoro-3-(N-(3-fluorophenyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide

Compound 45 (yield 9%) was synthesized using the same method as the step B synthesis of compound 26 above.

^1H NMR (300 MHz, DMSO _-d6 ) δ 13.36 (s, 1H), 11.01 (s, 1H), 10.76 (s, 1H), 8.76 (d, J = 2.2 Hz, 1H), 8.57 (d, J = 2.3 Hz, 1H), 8.13-7.97 (m, 2H), 7.62-7.52 (m, 1H), 7.30 (q, J = 7.8 Hz, 1H), 7.05-6.93 (m, 2H), 6.91-6.83 (m, 1H), 2.14 (s, 3H).

Compound 46. N-(5-(2-fluoro-5-(N-(3-fluorophenyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide

Compound 46 (yield 8%) was synthesized using the same method as the step B synthesis method for compound 26 above.

^1H NMR (300 MHz, DMSO _-d6 ) δ 13.43 (s, 1H), 10.81 (s, 1H), 10.65 (s, 1H), 8.61 (t, J = 2.1 Hz, 1H), 8.54 (s, 1H), 7.97 (dd, J = 7.2, 2.4 Hz, 1H), 7.84 (ddd, J = 7.3, 4.4, 2.4 Hz, 1H), 7.58 (dd, J = 10.2, 8.7 Hz, 1H), 7.31 (td, J = 8.3, 6.7 Hz, 1H), 7.05-6.83 (m, 3H), 2.14 (s, 3H).

Compound 47. N-(5-(3-fluoro-5-(3-fluorophenyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide

Compound 47 (yield 18%) was synthesized using the same method as the step B synthesis of compound 26 above.

^1H NMR (300 MHz, DMSO _- d6 ) δ 13.42 (s, 1H), 10.83-10.68 (m, 2H), 8.79 (d, J = 2.3 Hz, 1H), 8.62 (d, J = 2.3 Hz, 1H), 7.98-7.88 (m, 2H), 7.57 (dt, J = 7.9, 2.0 Hz, 1H), 7.31 (td, J = 8.4, 6.7 Hz, 1H), 7.03-6.86 (m, 3H), 2.16 (s, 3H).

Compound 48. N-(5-(2,4-difluoro-5-(N-(3-fluorophenyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide

Compound 48 (yield 20%) was synthesized using the same method as the step B synthesis of compound 26 above.

^1H NMR (300 MHz, DMSO _-d6 ) δ 13.42 (s, 1H), 11.04 (s, 1H), 10.79 (s, 1H), 8.61 (d, J = 2.0 Hz, 1H), 8.52 (s, 1H), 8.02 (t, J = 8.1 Hz, 1H), 7.74 (t, J = 10.2 Hz, 1H), 7.32 (q, J = 7.7 Hz, 1H), 7.05-6.84 (m, 3H), 2.13 (s, 3H).

Compound 49. N-(5-(2-fluoro-3-(N-(3-fluorophenyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide

Compound 49 (yield 23%) was synthesized using the same method as the step B synthesis of compound 26 above.

^1H NMR (300 MHz, DMSO _-d6 ) δ 13.41 (s, 1H), 11.00 (s, 1H), 10.79 (s, 1H), 8.59 (s, 1H), 8.55 (s, 1H), 7.96-7.86 (m, 2H), 7.50 (t, J = 7.8 Hz, 1H), 7.29 (q, J = 8.0 Hz, 1H), 7.01-6.82 (m, 3H), 2.12 (s, 3H).

Compound 50. N-(5-(5-(N-(4-fluorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide

Compound 50 (yield 8%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (400 MHz, Methanol-d ₄ ) δ 8.51 (s, 1H), 8.41 (s, 1H), 7.73 (dd, J = 8.7, 2.3 Hz, 1H), 7.66 (d, J = 2.3 Hz, 1H), 7.20 (d, J = 8.8 Hz, 1H), 7.16 - 7.10 (m, 2H), 7.00 (t, J = 8.7 Hz, 2H), 3.89 (s, 3H), 2.25 (s, 3H).

Compound 51. N-(5-(5-(N-(2-fluorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide

Compound 51 (yield 15%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.31 (s, 1H), 10.73 (s, 1H), 10.08 (s, 1H), 8.51 (d, J = 2.0 Hz, 1H), 8.36 (s, 1H), 7.72 (dd, J = 8.7, 2.4 Hz, 1H), 7.64 (d, J = 2.4 Hz, 1H), 7.34 - 7.27 (m, 2H), 7.17 (t, J = 7.9 Hz, 3H), 3.86 (s, 3H), 2.14 (s, 3H).

Compound 52. N-(5-(5-(N-(3-chlorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide

Compound 52 (yield 41%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 10.72 (s, 1H), 10.47 (s, 1H), 8.52 (d, J = 2.1 Hz, 1H), 8.37 (d, J = 2.1 Hz, 1H), 7.79 (dd, J = 8.7, 2.4 Hz, 1H), 7.71 (d, J = 2.4 Hz, 1H), 7.29 (t, J = 8.1 Hz, 2H), 7.19 - 7.05 (m, 3H), 3.84 (s, 3H), 2.12 (d, J = 5.9 Hz, 3H).

Compound 53. N-(5-(5-(N-(4-chlorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide

Compound 53 (yield 25%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 10.74 (s, 1H), 10.34 (s, 1H), 8.52 (d, J = 2.1 Hz, 1H), 8.38 (d, J = 2.2 Hz, 1H), 7.77 - 7.67 (m, 2H), 7.37 - 7.24 (m, 3H), 7.19 - 7.11 (m, 2H), 3.83 (s, 3H), 2.14 (s, 3H).

Compound 54. N-(5-(5-(N-(2-chlorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide

Compound 54 (yield 35%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 10.71 (s, 1H), 9.90 (s, 1H), 8.51 (d, J = 2.1 Hz, 1H), 8.36 (d, J = 2.2 Hz, 1H), 7.72 (dd, J = 8.7, 2.4 Hz, 1H), 7.63 (d, J = 2.4 Hz, 1H), 7.43 (dd, J = 7.8, 1.1 Hz, 1H), 7.34 - 7.26 (m, 3H), 7.20 (ddd, J = 7.9, 5.2, 3.8 Hz, 1H), 3.86 (s, 3H), 2.13 (s, 3H).

Compound 55. N-(5-(5-(N-(3-hydroxyphenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide

Compound 55 (yield 15%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.31 (s, 1H), 10.73 (s, 1H), 10.05 (s, 1H), 9.49 - 9.36 (m, 1H), 8.50 (d, J = 9.3 Hz, 1H), 8.39 (d, J = 2.1 Hz, 1H), 7.79 - 7.64 (m, 2H), 7.35 - 7.21 (m, 1H), 7.07 - 6.97 (m, 1H), 6.67 - 6.49 (m, 2H), 6.41 (d, J = 8.4 Hz, 1H), 3.90 - 3.80 (m, 3H), 2.18 - 2.09 (m, 3H).

Compound 56. N-(5-(5-(N-(4-hydroxyphenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide

Compound 56 (yield 7%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.31 (s, 1H), 10.74 (s, 1H), 9.64 (s, 1H), 9.32 (s, 1H), 8.51 (d, J = 2.1 Hz, 1H), 8.39 (d, J = 2.2 Hz, 1H), 7.66 - 7.60 (m, 2H), 7.25 (d, J = 9.4 Hz, 1H), 6.90 (d, J = 8.8 Hz, 2H), 6.64 (d, J = 8.7 Hz, 2H), 3.84 (s, 3H), 2.14 (s, 3H).

Compound 57. N-(5-(2-methoxy-5-(N-(pyridin-3-yl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide

Compound 57 (yield 14%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (400 MHz, DMSO-d ₆ ) δ 10.73 (s, 1H), 10.44 (s, 1H), 8.53 (d, J = 1.9 Hz, 1H), 8.38 (s, 1H), 8.29 (d, J = 19.3 Hz, 2H), 7.77 (dd, J = 8.8, 2.4 Hz, 1H), 7.71 (s, 1H), 7.56 (d, J = 7.2 Hz, 1H), 7.30 (d, J = 8.7 Hz, 2H), 3.86 (s, 3H), 2.15 (s, 3H).

Compound 58. N-(5-(2-methoxy-5-(N-(pyridin-2-yl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide

Compound 58 (yield 15%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (400 MHz, DMSO-d ₆ ) δ 10.73 (s, 1H), 8.58 (d, J = 1.8 Hz, 1H), 8.41 (s, 1H), 8.07 (s, 1H), 7.90 (d, J = 9.2 Hz, 1H), 7.84 (s, 1H), 7.70 (s, 1H), 7.29 (d, J = 8.7 Hz, 1H), 7.14 (s, 1H), 6.88 (s, 1H), 3.85 (s, 3H), 2.14 (d, J = 9.0 Hz, 3H).

Compound 59. N-(5-(2-methoxy-5-(morpholinosulfonyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide

Compound 59 (yield 64%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, Chloroform-d) δ 12.68 (s, 1H), 9.64 (s, 1H), 8.76 (d, J = 2.0 Hz, 1H), 8.67 (d, J = 2.1 Hz, 1H), 7.77 (dq, J = 4.3, 2.4 Hz, 2H), 7.11 (d, J = 9.3 Hz, 1H), 3.92 (s, 3H), 3.76 (dd, J = 5.7, 3.7 Hz, 4H), 3.12 - 2.98 (m, 4H), 2.32 (s, 3H).

Compound 60. N-(5-(5-(N-(3,5-difluorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide

Compound 60 (yield 31%) was synthesized using the same method as the synthesis of compound 24 above.

^1H NMR (400 MHz, DMSO _-d6 ) δ 13.31 (s, 1H), 10.79 (s, 1H), 10.73 (s, 1H), 8.54 (d, J = 2.2 Hz, 1H), 8.37 (d, J = 2.1 Hz, 1H), 7.85 (dd, J = 8.8, 2.5 Hz, 1H), 7.75 (d, J = 2.5 Hz, 1H), 7.32 (d, J = 8.9 Hz, 1H), 6.94-6.84 (m, 1H), 6.84-6.76 (m, 2H), 3.85 (s, 3H), 2.12 (s, 3H).

Compound 61. N-(5-(5-(N-(3-chloro-5-fluorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide

Compound 61 (yield 36%) was synthesized using the same method as the synthesis of compound 24 above.

^1H NMR (400 MHz, DMSO _-d6 ) δ 13.31 (s, 1H), 10.79 (s, 1H), 10.73 (s, 1H), 8.53 (d, J = 2.2 Hz, 1H), 8.37 (d, J = 2.2 Hz, 1H), 7.84 (dd, J = 8.7, 2.5 Hz, 1H), 7.74 (d, J = 2.5 Hz, 1H), 7.33 (d, J = 8.8 Hz, 1H), 7.09 (d, J = 8.6 Hz, 1H), 7.02-6.90 (m, 2H), 3.85 (s, 3H), 2.12 (s, 3H).

Compound 62. N-(3-fluorophenyl)-4-methoxy-3-(3-phenyl-1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide

62-1. Synthesis of 5-bromo-3-iodo-1H-pyrazolo[3,4-b]pyridine (Compound 62-1)

3-Bromo-1H-pyrazolo[3,4-b]pyridine (9.9 g, 50.0 mmol) and DMF (50 mL) were added at 0 ℃, potassium hydroxide (11.2 g, 200.0 mmol)(4.0 eq.) was added, and the mixture was stirred for 10 minutes. Iodine (14.0 g, 55.0 mmol)(1.2 eq.) was added, and the mixture was stirred at room temperature for 20 hours. After completion of the reaction, sat. sodium thiosulfate and water were added, and the resulting solid was filtered to obtain compound 62-1 in a yield of 100% (16.2 g).

62-2. Synthesis of 5-bromo-3-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridine (Compound 62-2)

Compound 62-1 (5-bromo-3-iodo-1H-pyrazolo[3,4-b]pyridine; 16.2 g, 50.0 mmol) and DMF (200 mL) were added and stirred at 0 °C. Then, sodium hydride (4.0 mg, 100.0 mmol) (2.0 eq.) and 2-(Trimethylsilyl)ethoxymethyl chloride (15 mL, 85.0 mmol) (1.7 eq.) were added and stirred at room temperature for 18 h. After completion of the reaction, the mixture was quenched with water, extracted with EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. Compound 62-2 was obtained in a yield of 60% (13.6 g) through separation by silica gel column chromatography.

62-3. Synthesis of 5-bromo-3-phenyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridine (Compound 62-3)

Compound 62-2 (5-bromo-3-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridine; 1.36 g, 3.0 mmol), phenylboronic acid (402 mg, 3.3 mmol)(1.1 eq.), [1,1′-Bis(diphenylphosphino)ferrocene]dichloropalladium(II) dichloromethane complex (122 mg, 0.15 mmol)(0.05 eq.), potassium carbonate aqueous solution (2 M, 4.5 mL)(3.0 eq.), and CH ₃ CN (10 mL) were added and stirred at 85 ℃ for 6 h. After completion of the reaction, the mixture was filtered using a celite pad, extracted with water and EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. Afterwards, the compound 62-3 was separated by silica gel column chromatography, and a yield of 60% (728 mg) was obtained.

62-4. Synthesis of 5-bromo-3-phenyl-1H-pyrazolo[3,4-b]pyridine (Compound 62-4)

Compound 62-3 (5-bromo-3-phenyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-b]pyridine; 728 mg, 1.8 mmol) and THF (10 mL) were stirred, and tetrabutylammonium fluoride (1 M in THF, 18 mL) (20.0 eq.) was added. The mixture was refluxed at 85°C for 24 h. After completion of the reaction, the solvent was removed, extracted with water and EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. Compound 62-4 was obtained in a yield of 83% (410 mg) by separation using silica gel column chromatography.

62-5. Synthesis of N-(3-fluorophenyl)-4-methoxy-3-(3-phenyl-1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 62)

Compound 62 (yield 40%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.94 (s, 1H), 10.46 (s, 1H), 8.60 (d, J = 2.0 Hz, 1H), 8.47 (d, J = 2.0 Hz, 1H), 8.08 - 7.99 (m, 2H), 7.84 (dd, J = 6.6, 2.5 Hz, 2H), 7.61 - 7.51 (m, 2H), 7.51 - 7.39 (m, 1H), 7.37 - 7.22 (m, 2H), 7.03 - 6.91 (m, 2H), 6.91 - 6.79 (m, 1H), 3.85 (s, 3H).

Compound 63. N-(3-fluorophenyl)-4-methoxy-3-(3-(thiophen-3-yl)-1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide

Compound 62 was synthesized using the same method as in Synthetic Methods 62-1 to 62-4, and then compound 63 (yield 12%) was synthesized using the same method as in Synthetic Method 1.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 13.80 (s, 1H), 10.46 (s, 1H), 8.61 - 8.52 (m, 2H), 8.22 (dd, J = 2.7, 1.5 Hz, 1H), 7.83 (h, J = 2.5 Hz, 2H), 7.79 - 7.68 (m, 2H), 7.37 - 7.22 (m, 2H), 7.02 - 6.92 (m, 2H), 6.90 - 6.82 (m, 1H), 3.85 (s, 3H).

Compound 64. 4-Methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide

64-1. Synthesis of 4-(5-bromo-1H-pyrazolo[3,4-b]pyridin-3-yl)morpholine (compound 44-1)

5-Bromo-1H-pyrazolo[3,4-b]pyridin-3-amine (9.0 g, 42.25 mmol), 2-bromoethyl ether (9.0 mL, 71.83 mmol)(1.7 eq.), and N,N-diisopropylethylamine (15 mL, 86.61 mmol)(2.05 eq.) were added to DMF (42 mL) and stirred at 110 °C for 2 h. After completion of the reaction, the mixture was extracted with water and EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain compound 64-1 in a yield of 35% (4.19 g).

64-2. Synthesis of 4-methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide (Compound 64)

Compound 64 (yield 54%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, Chloroform-d) δ 8.43 (d, J = 1.9 Hz, 1H), 8.05 (d, J = 1.9 Hz, 1H), 7.88 (s, 1H), 7.83 - 7.72 (m, 2H), 7.10 (t, J = 7.7 Hz, 1H), 6.93 (dd, J = 14.7, 8.3 Hz, 4H), 3.93 - 3.84 (m, 4H), 3.82 (s, 3H), 3.47 - 3.37 (m, 4H), 2.25 (s, 3H).

Compound 65. 4-Methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(p-tolyl)benzenesulfonamide

Compound 65 (yield 20%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, Chloroform-d) δ 8.49 (d, J = 1.8 Hz, 1H), 8.11 (d, J = 1.9 Hz, 1H), 7.80 (d, J = 2.3 Hz, 1H), 7.76 (dd, J = 8.6, 2.4 Hz, 1H), 7.70 (s, 1H), 7.12-7.06 (m, 4H), 6.98 (d, J = 8.6 Hz, 1H), 3.99 - 3.89 (m, 4H), 3.86 (s, 3H), 3.55 - 3.41 (m, 4H), 2.29 (s, 3H).

Compound 66. 4-Methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(o-tolyl)benzenesulfonamide

Compound 66 (yield 12%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, Chloroform-d) δ 8.53 (d, J = 4.0 Hz, 1H), 8.11 (d, J = 8.0 Hz, 1H), 7.77 (dd, J = 8.7, 2.3 Hz, 1H), 7.36 - 7.30 (m, 1H), 7.20 - 7.13 (m, 4H), 7.10 (s, 1H), 7.03 - 6.98 (m, 1H), 3.98 - 3.90 (m, 4H), 3.88 (s, 3H), 3.53 - 3.43 (m, 4H), 2.11 (s, 3H).

Compound 67. 4-Methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)-N-phenylbenzenesulfonamide

Compound 67 (yield 24%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, Chloroform-d) δ 8.49 (s, 1H), 8.09 (s, 1H), 7.86 - 7.74 (m, 2H), 7.69 (s, 1H), 7.26 (s, 1H), 7.22 - 7.11 (m, 3H), 7.00 (d, J = 8.5 Hz, 1H), 3.94 (q, J = 5.0 Hz, 4H), 3.87 (s, 3H), 3.56 - 3.42 (m, 4H).

Compound 68. N-(3-fluorophenyl)-4-methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide

Compound 68 (yield 13%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 12.73 (brs, 1H), 10.44 (brs, 1H), 8.42 (s, 1H), 8.24 (d, J = 2.5 Hz, 1H), 7.85 - 7.71 (m, 2H), 7.34 - 7.22 (m, 2H), 6.98 - 6.80 (m, 3H), 3.89 - 3.73 (m, 7H), 3.37 (m, 4H).

Compound 69. N-(2-fluorophenyl)-4-methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide

Compound 69 (yield 21%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 12.73 (s, 1H), 10.06 (s, 1H), 8.43 (d, J = 2.0 Hz, 1H), 8.21 (s, 1H), 7.73 (dd, J = 8.8, 2.4 Hz, 1H), 7.66 (d, J = 2.3 Hz, 1H), 7.36 - 7.22 (m, 2H), 7.22 - 7.01 (m, 3H), 3.84 (s, 3H), 3.83 - 3.74 (m, 4H), 3.48 - 3.38 (m, 4H).

Compound 70. N-(2-chlorophenyl)-4-methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide

Compound 70 (yield 54%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, Chloroform-d) δ 10.64 (s, 1H), 8.47 (d, J = 1.9 Hz, 1H), 8.02 (d, J = 1.9 Hz, 1H), 7.80 (dd, J = 8.7, 2.5 Hz, 1H), 7.74 - 7.67 (m, 2H), 7.31 - 7.27 (m, 1H), 7.12 - 7.05 (m, 2H), 6.99 (d, J = 8.8 Hz, 1H), 3.97 - 3.90 (m, 4H), 3.86 (s, 3H), 3.47 (dd, J = 5.9, 3.7 Hz, 4H).

Compound 71. N-(3-hydroxyphenyl)-4-methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide

Compound 71 (yield 23%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 12.73 (s, 1H), 10.03 (s, 1H), 9.45 (s, 1H), 8.43 (d, J = 2.0 Hz, 1H), 8.23 (d, J = 2.0 Hz, 1H), 7.77 (dd, J = 8.7, 2.4 Hz, 1H), 7.70 (d, J = 2.4 Hz, 1H), 7.28 (d, J = 8.8 Hz, 1H), 7.00 (t, J = 8.1 Hz, 1H), 6.63 (t, J = 2.2 Hz, 1H), 6.58 - 6.53 (m, 1H), 6.44 - 6.38 (m, 1H), 3.83 (s, 3H), 3.80 (t, J = 4.8 Hz, 4H), 3.42 (s, 4H).

Compound 72. 4-((4-methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)phenyl)sulfonyl)morpholine

Compound 72 (yield 64%) was synthesized using the same method as the synthesis method for compound 1 above.

^1H NMR (300 MHz, Chloroform-d) δ 12.68 (s, 1H), 9.64 (s, 1H), 8.76 (d, J = 2.0 Hz, 1H), 8.67 (d, J = 2.1 Hz, 1H), 7.77 (dq, J = 4.3, 2.4 Hz, 2H), 7.11 (d, J = 9.3 Hz, 1H), 3.92 (s, 3H), 3.76 (dd, J = 5.7, 3.7 Hz, 4H), 3.12 - 2.98 (m, 4H), 2.32 (s, 3H).

Compound 73. 4-Methoxy-3-(1-phenyl-1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide

4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide (99 mg, 0.25 mmol), phenylboronic acid (46 mg, 0.375 mmol)(1.5 eq.), copper(II) acetate (14 mg, 0.075 mmol)(0.3 eq.), pyridine (60 μL, 0.75 mmol)(3.0 eq.), and DMF (3 mL) were added and stirred at 90 ℃ for 12 h. After completion of the reaction, the mixture was filtered using a celite pad, extracted with water and EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain compound 73 in a yield of 61% (72 mg).

^1H NMR (300 MHz, Chloroform-d) δ 8.72 (d, J = 2.1 Hz, 1H), 8.29 - 8.20 (m, 3H), 8.14 (d, J = 2.0 Hz, 1H), 7.82 (dd, J = 8.6, 2.4 Hz, 1H), 7.78 (d, J = 2.3 Hz, 1H), 7.61 - 7.50 (m, 2H), 7.39 - 7.31 (m, 1H), 7.15 (t, J = 7.7 Hz, 1H), 7.02 (d, J = 8.7 Hz, 1H), 6.99 - 6.90 (m, 3H), 3.87 (s, 3H), 2.29 (s, 3H).

Compound 74. 3-(1-(2-(dimethylamino)ethyl)-1H-pyrazolo[3,4-b]pyridin-5-yl)-4-methoxy-N-(m-tolyl)benzenesulfonamide

4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide (99 mg, 0.25 mmol), 2-chloro-NN-dimethylethylamine-hydrochloride (54 mg, 0.375 mmol)(1.5 eq.), Cs ₂ CO ₃ (244 mg, 0.75 mmol)(3.0 eq.) and DMF (3 mL) were added and stirred at 65 ℃ for 24 h. After completion of the reaction, the mixture was filtered using a celite pad, extracted with water and EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain compound 74 in a yield of 30% (35 mg).

¹ H NMR 300 MHz, Chloroform-d)) δ 8.63 (d, J = 1.9 Hz, 1H), 8.13 (d, J = 2.1 Hz, 2H), 7.68 (dd, J = 8.7, 2.2 Hz, 1H), 7.54 (d, J = 2.2 Hz, 1H), 7.22 (t, J = 7.7 Hz, 1H), 7.14 (d, J = 7.5 Hz, 1H), 7.05 (d, J = 8.7 Hz, 1H), 7.00 (s, 1H), 6.87 (d, J = 8.0 Hz, 1H), 3.92 (s, 3H), 3.75 - 3.63 (m, 2H), 2.52 - 2.39 (m, 2H), 2.32 (s, 3H), 2.23 (s, 6H).

Compound 75. 4-Methoxy-3-(1-methyl-1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide

75-1. Synthesis of 5-bromo-1-methyl-1H-pyrazolo[3,4-b]pyridine (Compound 75-1)

3-Bromo-1H-pyrazolo[3,4-b]pyridine (408 mg, 2.0 mmol), potassium carbonate (553 mg, 4.0 mmol)(2.0 eq.) and DMF (10 mL) were stirred, then iodomethane (150 μL, 2.4 mmol)(1.2 eq.) was added and stirred at room temperature for 12 h. After completion of the reaction, the mixture was extracted with water and EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain compound 75-1 in a yield of 49% (206 mg).

75-2. Synthesis of 5-(2-methoxyphenyl)-1-methyl-1H-pyrazolo[3,4-b]pyridine (Compound 75-2)

Compound 75-1 (5-bromo-1-methyl-1H-pyrazolo[3,4-b]pyridine; 205 mg, 0.96 mmol), 2-methoxyphenylboronic acid (166 mg, 1.06 mmol)(1.1 eq.), [1,1′-Bis(diphenylphosphino)ferrocene]dichloropalladium(II) dichloromethane complex (42 mg, 0.05 mmol)(0.05 eq.), potassium carbonate (2.0 M in H ₂ O, 3 mL), and CH ₃ CN (3 mL) were added and refluxed for 4 h. After completion of the reaction, the solvent was removed by celite filtration, extracted with water and EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. Compound 75-2 was obtained in a yield of 79% (181 mg) through separation by silica gel column chromatography.

75-3. Synthesis of 4-methoxy-3-(1-methyl-1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonyl chloride (Compound 75-3)

Chlorosulfonic acid (1 mL) was slowly added to compound 75-2 (5-(2-methoxyphenyl)-1-methyl-1H-pyrazolo[3,4-b]pyridine; 181 mg, 0.76 mmol) at 0 °C and stirred for 2 hours. After completion of the reaction, ice water was slowly added, extracted with EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. Compound 75-3 was obtained in a yield of 86% (221 mg) by separation using silica gel column chromatography.

75-4. 4-methoxy-3-(1-methyl-1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)

Synthesis of benzenesulfonamide (compound 75)

Compound 75-3 (4-methoxy-3-(1-methyl-1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonyl chloride; 220 mg, 0.65 mmol), m-toluidine (110 μL, 0.98 mmol)(1.5 eq.), and pyridine (55 μL, 0.65 mmol)(1.0 eq.) were added to THF (2 mL), and the mixture was stirred at room temperature for 12 h. After completion of the reaction, the mixture was extracted with water and EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain compound 75 in a yield of 99% (262 mg).

¹ H NMR 300 MHz, Chloroform-d) δ 8.56 (d, J = 2.0 Hz, 1H), 8.01 - 7.96 (m, 2H), 7.81 (dd, J = 8.7, 2.4 Hz, 1H), 7.74 (d, J = 2.4 Hz, 1H), 7.37 (s, 1H), 7.17 - 7.08 (m, 1H), 7.00 - 6.90 (m, 4H), 4.16 (s, 3H), 3.83 (s, 3H), 2.27 (s, 3H).

Compound 76. 3-(1-Acetyl-1H-pyrazolo[3,4-b]pyridin-5-yl)-4-methoxy-N-(m-tolyl)benzenesulfonamide

76-1. Synthesis of 1-(5-bromo-1H-pyrazolo[3,4-b]pyridin-1-yl)ethan-1-one (Compound 76-1)

3-bromo-1H-pyrazolo[3,4-b]pyridine (1.98 g, 10.0 mmol), DMAP (611 mg, 5.0 mmol)(0.5 eq.), THF (300 mL), Et ₃ N (4.5 mL, 30.0 mmol)(3.0 eq.) were added and stirred at 0 ℃, then acetic anhydride (1.9 mL, 20.0 mmol)(2.0 eq.) was added and stirred at 80 ℃ for 12 h. After the reaction was completed, the mixture was extracted with water and DCM, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. After that, it was filtered with MeOH to obtain compound 76-1 with a yield of 88% (2.13 g).

76-2. Synthesis of 3-(1-acetyl-1H-pyrazolo[3,4-b]pyridin-5-yl)-4-methoxy-N-(m-tolyl)benzenesulfonamide (Compound 76)

3-Bromo-4-methoxy-N-(m-tolyl)benzenesulfonamide (712 mg, 2.0 mmol), Bis(pinacolato)diboron (609 mg, 2.4 mmol)(1.2 eq.), [1,1′-Bis(diphenylphosphino)ferrocene]dichloropalladium(II) dichloromethane complex (163 mg, 0.2 mmol)(10 mol%), potassium acetate (589 mg, 6.0 mmol)(3.0 eq.), and 1,4-Dioxane (10 mL) were added, degassing was performed under N ₂ for 10 min, and the mixture was stirred at 100 °C for 12 h. After the reaction was completed, the solvent was removed by celite filtration, and then compound 76-1 (1-(5-bromo-1H-pyrazolo[3,4-b]pyridin-1-yl)ethan-1-one; 480 mg, 2.0 mmol)(1.0 eq.), Bis(diphenylphosphino)ferrocene]dichloropalladium (146 mg, 0.2 mmol)(10 mol%), potassium carbonate aqueous solution (2.0 M, 2 mL)(2.0 eq.), and 1,4-Dioxane (5 mL) were added and stirred at room temperature for 12 hours. After the reaction was completed, the solvent was removed by celite filtration, and the mixture was extracted with water and EtOAc. The organic layer was dried over MgSO ₄ and concentrated under reduced pressure. The residue was separated by silica gel column chromatography to obtain compound 76 in a yield of 1% (10 mg).

^1H NMR (300 MHz, Chloroform-d) δ 8.78 (d, J = 1.8 Hz, 1H), 8.16 (s, 1H), 8.06 (d, J = 2.0 Hz, 1H), 7.84 (dd, J = 8.6, 2.2 Hz, 1H), 7.77 (d, J = 2.2 Hz, 1H), 7.33 (s, 1H), 7.12 (t, J = 7.7 Hz, 1H), 6.97 (dd, J = 16.1, 8.5 Hz, 3H), 6.91 (s, 1H), 3.84 (s, 3H), 2.88 (s, 3H), 2.26 (s, 3H).

Compound 77. N-((3-(1-Acetyl-1H-pyrazolo[3,4-b]pyridin-5-yl)-4-methoxyphenyl)sulfonyl)-N-(m-tolyl)acetamide

3-Bromo-4-methoxy-N-(m-tolyl)benzenesulfonamide (99 mg, 0.25 mmol), THF (3 mL), and triethylamine (100 μL, 0.75 mmol) were stirred at 0 °C, and acetic anhydride (120 μL, 1.25 mmol) was slowly added. The mixture was stirred at 80 °C for 12 h. After completion of the reaction, the mixture was extracted with water and EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. Compound 77 was obtained in a yield of 76% (83 g) through separation by silica gel column chromatography.

^1H NMR (300 MHz, Chloroform-d) δ 8.95 (d, J = 2.0 Hz, 1H), 8.27 - 8.19 (m, 2H), 8.11 (dd, J = 8.7, 2.4 Hz, 1H), 8.06 (d, J = 2.4 Hz, 1H), 7.32 (dd, J = 16.6, 7.6 Hz, 2H), 7.15 (d, J = 8.8 Hz, 2H), 7.04 (d, J = 7.7 Hz, 1H), 3.93 (s, 3H), 2.91 (s, 3H), 2.40 (s, 3H), 1.87 (s, 3H).

Compound 78. 4-((3-(1H-indazol-5-yl)-4-methoxyphenyl)sulfonyl)morpholine

4-((3-bromo-4-methoxyphenyl)sulfonyl)morpholine (200 mg, 0.6 mmol), bis(pinacolato)diboron (183 mg, 0.72 mmol), [1,1′-Bis(diphenylphosphino)ferrocene]dichloropalladium(II) dichloromethane complex (49 mg, 0.06 mmol)(10 mol%), potassium acetate (177 mg, 1.8 mmol), and 1,4-dioxane (2 mL) were added and reacted using a microwave reactor at 120 ℃ for 1 h. After cooling to room temperature, 5-bromo-1H-indazole (118 mg, 0.6 mmol) and sodium carbonate aqueous solution (2 M, 0.5 mL) were added and reacted using a microwave reactor at 120 ℃ for 1 h. After the reaction was completed, the solvent was removed by celite filtration, extracted with water and EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. After separation by silica gel column chromatography, compound 78 was obtained with a yield of 52% (117 mg).

¹ H NMR (300 MHz, Chloroform-d) δ 10.36 (s, 1H), 8.14 (s, 1H), 7.89 (t, J = 1.2 Hz, 1H), 7.78 - 7.72 (m, 2H), 7.56 (d, J = 1.2 Hz, 2H), 7.14 - 7.08 (m, 1H), 3.92 (s, 3H), 3.79 - 3.73 (m, 4H), 3.07 - 3.00 (m, 4H).

Compound 79. 3-(1H-indazol-5-yl)-4-methoxy-N-(m-tolyl)benzenesulfonamide

Compound 79 (yield 59%) was synthesized using the same method as the step B synthesis method of compound 26 above.

^1H NMR (300 MHz, DMSO _-d6 ) δ 13.14 (s, 1H), 10.05 (s, 1H), 8.12 (t, J = 1.2 Hz, 1H), 7.75-7.70 (m, 2H), 7.65 (d, J = 2.4 Hz, 1H), 7.58 (dt, J = 8.7, 1.0 Hz, 1H), 7.38 (dd, J = 8.7, 1.6 Hz, 1H), 7.24 (d, J = 8.8 Hz, 1H), 7.13 (t, J = 7.7 Hz, 1H), 7.00-6.90 (m, 2H), 6.90-6.85 (m, 1H), 3.82 (s, 3H), 2.21 (s, 3H).

Compound 80. N-(3-fluorophenyl)-3-(1H-indazol-5-yl)-4-methoxybenzenesulfonamide

Compound 80 (yield 33%) was synthesized using the same method as the step B synthesis of compound 26 above.

^1H NMR (300 MHz, DMSO _-d6 ) δ 13.11 (s, 1H), 10.39 (s, 1H), 8.10 (t, J = 1.2 Hz, 1H), 7.77-7.72 (m, 2H), 7.70 (d, J = 2.4 Hz, 1H), 7.57 (dt, J = 8.6, 1.0 Hz, 1H), 7.39 (dd, J = 8.7, 1.6 Hz, 1H), 7.32-7.21 (m, 2H), 6.99-6.91 (m, 2H), 6.88-6.80 (m, 1H), 3.83 (s, 3H).

Compound 81. 2,4-Difluoro-N-(3-fluorophenyl)-5-(1H-indazol-5-yl)benzenesulfonamide

Compound 81 (yield 48%) was synthesized using the same method as the step B synthesis of compound 26 above.

^1H NMR (300 MHz, DMSO _-d6 ) δ 13.25 (s, 1H), 11.02 (s, 1H), 8.17 (d, J = 1.5 Hz, 1H), 8.03-7.90 (m, 2H), 7.73-7.63 (m, 2H), 7.47 (dt, J) = 8.8, 1.8 Hz, 1H), 7.32 (q, J = 7.7 Hz, 1H), 7.04-6.84 (m, 3H).

Compound 82. N-(5-(5-(N-(3-fluorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-indazol-3-yl)acetamide

Step A) Preparation of N-(5-bromo-1H-indazol-3-yl)acetamide

5-Bromo-1H-indazole-3-amine (440 mg, 2.07 mmol) and pyridine (10 mL) were stirred at 0 °C, then acetyl chloride (160 μL, 2.28 mmol) was slowly added and stirred at room temperature for 12 h. After completion of the reaction, the mixture was extracted with water and EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain N-(5-bromo-1H-indazol-3-yl)acetamide in a yield of 58% (308 mg).

Step B) Preparation of N-(5-(5-(N-(3-fluorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-indazol-3-yl)acetamide (Compound 82)

Compound 82 (yield 3%) was synthesized using the same method as the step B synthesis of compound 26 above.

^1H NMR (300 MHz, DMSO -d6 ) δ 12.72 (s, 1H), 10.47 ( _s , 1H), 10.41 (s, 1H), 7.79-7.73 (m, 2H), 7.66 (d, J = 2.4 Hz, 1H), 7.47 (d, J = 8.7) Hz, 1H), 7.38 (d, J = 8.7 Hz, 1H), 7.34-7.23 (m, 2H), 6.99-6.81 (m, 3H), 3.81 (s, 3H), 2.11 (s, 3H).

Compound 83. N-(5-(2,4-difluoro-5-(N-(3-fluorophenyl)sulfamoyl)phenyl)-1H-indazol-3-yl)acetamide

Compound 83 (yield 18%) was synthesized using the same method as the step B synthesis of compound 26 above.

^1H NMR (400 MHz, DMSO _-d6 ) δ 12.83 (s, 1H), 11.02 (s, 1H), 10.48 (s, 1H), 7.96-7.88 (m, 2H), 7.67 (t, J = 10.2 Hz, 1H), 7.55 (d, J = 8.7 Hz, 1H), 7.45 (d, J = 8.8 Hz, 1H), 7.34-7.27 (m, 1H), 7.01-6.92 (m, 2H), 6.88 (td, J = 8.5, 2.5 Hz, 1H), 2.12 (s, 3H).

Compound 84. 3-(1-Acetyl-3-amino-1H-indazol-5-yl)-N-(3-fluorophenyl)-4-methoxybenzenesulfonamide

Step A) Preparation of 1-(3-amino-5-bromo-1H-indazol-1-yl)ethan-1-one

5-Bromo-1H-indazole-3-amine (1.06 g, 5.0 mmol), triethylamine (1.2 mL, 15 mmol), THF (5 mL) were stirred at 0 ℃, and acetyl chloride (430 μL, 6.0 mmol) was slowly added, followed by stirring at room temperature for 12 h. After completion of the reaction, the mixture was extracted with water and EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain 1-(3-amino-5-bromo-1H-indazol-1-yl)ethan-1-one in a yield of 69% (880 mg).

Step B) Preparation of 3-(1-acetyl-3-amino-1H-indazol-5-yl)-N-(3-fluorophenyl)-4-methoxybenzenesulfonamide (Compound 84)

Compound 84 (yield 7%) was synthesized using the same method as the step B synthesis of compound 26 above.

^1H NMR (300 MHz, DMSO _- d6 ) δ 10.49 (s, 1H), 8.23 (d, J = 8.6 Hz, 1H), 8.04-7.98 (m, 1H), 7.79 (dd, J = 8.7, 2.5 Hz, 1H), 7.73 (d, J = 2.4) Hz, 1H), 7.57 (dd, J = 8.6, 1.7 Hz, 1H), 7.33-7.24 (m, 2H), 6.99-6.80 (m, 3H), 6.53 (s, 2H), 3.84 (s, 3H), 2.53 (s, 3H).

Compound 85. 5-(1-Acetyl-3-amino-1H-indazol-5-yl)-2,4-difluoro-N-(3-fluorophenyl)benzenesulfonamide

Compound 85 (yield 8%) was synthesized using the same method as the step B synthesis of compound 26 above.

^1H NMR (400 MHz, DMSO _-d6 ) δ 11.04 (s, 1H), 8.30 (d, J = 8.6 Hz, 1H), 8.13 (s, 1H), 8.02 (t, J = 8.1 Hz, 1H), 7.76-7.68 (m, 2H), 7.35-7.26 (m, 1H), 7.01-6.92 (m, 2H), 6.92-6.85 (m, 1H), 6.58 (s, 2H), 2.54 (s, 3H).

Compound 86. 4-((4-methoxy-3-(1H-pyrrolo[2,3-b]pyridin-5-yl)phenyl)sulfonyl)morpholine

4-((3-bromo-4-methoxyphenyl)sulfonyl)morpholine (200 mg, 0.6 mmol), bis(pinacolato)diboron (183 mg, 0.72 mmol), [1,1′-Bis(diphenylphosphino)ferrocene]dichloropalladium(II) dichloromethane complex (49 mg, 0.06 mmol), potassium acetate (177 mg, 1.8 mmol), and 1,4-dioxane (2 mL) were added and reacted using a microwave reactor at 120 ℃ for 1 h. After cooling to room temperature, 5-bromo-1H-pyrrolo[2,3-b]pyridine (118 mg, 0.6 mmol) and sodium carbonate aqueous solution (2 M, 0.5 mL) were added and reacted at 120 ℃ for 1 h. After the reaction was completed, the solvent was removed by celite filtration, extracted with water and EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. After separation by silica gel column chromatography, compound 86 was obtained with a yield of 67% (151 mg).

^1H NMR (400 MHz, DMSO-d ₆ ) δ 11.75 (s, 1H), 8.33 (d, J = 2.1 Hz, 1H), 8.09 (d, J = 2.1 Hz, 1H), 7.74 (dd, J = 8.7, 2.4 Hz, 1H), 7.61 (d, J = 8.7, 2.4 Hz, 1H) 2.4 Hz, 1H), 7.52 (dd, J = 3.4, 2.5 Hz, 1H), 7.39 (d, J = 8.7 Hz, 1H), 6.50 (dd, J = 3.4, 1.8 Hz, 1H), 3.90 (s, 3H), 3.64 (t, J = 4.6 Hz, 4H), 2.94 - 2.85 (m, 4H).

Compound 87. 4-Methoxy-3-(1H-pyrrolo[2,3-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide

87-1. Synthesis of 5-bromo-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine (Compound 87-1)

5-Bromo-1H-pyrrolo[2,3-b]pyridine (788 mg, 4.0 mmol) and THF (15 mL) were added, and sodium hydride (480 mg, 12.0 mmol) and benzyltriethylammonium chloride (18 mg, 0.08 mmol) were added at 0 °C, and the mixture was stirred for 30 min. Benzenesulfonyl chloride (615 μL, 4.8 mmol) was added, and the mixture was stirred at room temperature for 2 h. After the reaction was completed, the mixture was quenched with water, extracted with EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure to obtain compound 87-1 in a yield of 99% (1.35 g).

87-2. Synthesis of 5-(2-methoxyphenyl)-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine (Compound 87-2)

Compound 87-1 (1.35 g, 4.0 mmol), 2-methoxyphenylboronic acid (669 mg, 4.4 mmol), Tetrakis(triphenylphosphine)palladium (69 mg, 0.06 mmol), potassium carbonate (3.54 g, 25.6 mmol), and toluene/EtOH (30/10 mL) were added and stirred at 110 °C for 5 h. After completion of the reaction, the solvent was removed by celite filtration, extracted with water and EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. Compound 87-2 was obtained in a yield of 82% (1.2 g) through separation by silica gel column chromatography.

87-3. Synthesis of 5-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridine (Compound 87-3)

Compound 87-2 (1.09 mg, 3.0 mmol) and sodium tert-butoxide (576 mg, 6.0 mmol) were added to dioxane (10 mL) and stirred at 80 °C for 4 h. After completion of the reaction, the mixture was extracted with water and EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. Afterwards, the mixture was filtered with EtOAc and Hex to obtain compound 87-3 in a yield of 80% (535 mg).

87-4. Synthesis of (5-(2-methoxyphenyl)-1H-pyrrolo[2,3-b]pyridin-1-yl)(phenyl)methanone (Compound 87-4)

Compound 87-3 (527 mg, 2.35 mmol), triethylamine (3.2 mL, 23.5 mmol), 4-Dimethylaminopyridine (288 mg, 2.35 mmol), and THF (8 mL) were stirred, then benzoyl chloride (410 μL, 3.53 mmol) was added, and the mixture was stirred at room temperature for 12 h. After completion of the reaction, the mixture was extracted with water and EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. It was then filtered with EtOAc to obtain compound 87-4 in a yield of 49% (381 mg).

87-5. Synthesis of 4-methoxy-3-(1H-pyrrolo[2,3-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide (Compound 87)

Compound 87-4 (100 mg, 0.3 mmol) and DCM (1 mL) were stirred at 0 °C, and chlorosulfonic acid (30 μL, 0.45 mmol) was slowly added thereto and stirred for 1 h. After completion of the reaction, ice water was slowly added thereto, extraction was performed with EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. To this was added m-toluidine (50 μL, 0.45 mmol), pyridine (25 μL, 0.3 mmol), and THF (3 mL), and the mixture was stirred at room temperature for 12 h. After completion of the reaction, the mixture was extracted with water and EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. Compound 87 was obtained in a yield of 24% (28 mg, 2 steps) through separation by silica gel column chromatography.

^1H NMR 300 MHz, Chloroform-d)) δ 8.44 (s, 1H), 8.30 (s, 1H), 7.89 (s, 1H), 7.83 (d, J = 8.1 Hz, 1H), 7.51 (s, 1H), 7.14 (d, J = 7.8 Hz, 1H), 7.00 (dd, J = 12.9, 8.3 Hz, 4H), 6.69 (s, 1H), 3.88 (s, 3H), 2.30 (s, 3H).

Compound 88. N-(3-fluorophenyl)-4-methoxy-3-(1H-pyrrolo[2,3-b]pyridin-5-yl)benzenesulfonamide

Compound 88 (yield 67%) was synthesized using the same method as the step B synthesis of compound 26 above.

^1H NMR (300 MHz, DMSO _-d6 ) δ 11.75 (s, 1H), 10.43 (s, 1H), 8.21 (d, J = 2.1 Hz, 1H), 7.97 (d, J = 2.1 Hz, 1H), 7.77 (dd, J = 8.7, 2.5 Hz, 1H), 7.71 (d, J = 2.4 Hz, 1H), 7.58-7.44 (m, 1H), 7.35-7.23 (m, 2H), 7.00-6.92 (m, 2H), 6.92-6.82 (m, 1H), 6.50 (dd, J = 3.5, 1.8 Hz, 1H), 3.83 (s, 3H).

Compound 89. 2,4-Difluoro-N-(3-fluorophenyl)-5-(1H-pyrrolo[2,3-b]pyridin-5-yl)benzenesulfonamide

Compound 89 (yield 43%) was synthesized using the same method as the step B synthesis of compound 26 above.

^1H NMR (300 MHz, DMSO _-d6 ) δ 11.89 (s, 1H), 11.03 (s, 1H), 8.32 (s, 1H), 8.13 (s, 1H), 8.01 (t, J = 8.1 Hz, 1H), 7.72 (t, J = 10.3 Hz, 1H), 7.58 (t, J = 3.0 Hz, 1H), 7.33 (q, J = 7.7 Hz, 1H), 7.09-6.82 (m, 3H), 6.62-6.49 (m, 1H).

Compound 90. 4-Methoxy-3-(3-morpholino-1H-pyrrolo[2,3-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide

90-1. Synthesis of 5-bromo-3-nitro-1H-pyrrolo[2,3-b]pyridine (Compound 90-1)

Nitric acid (72.5 mL) was slowly added to 5-bromo-1H-pyrrolo[2,3-b]pyridine (10 g, 50.75 mmol) at 0 °C and stirred for 1 hour. After completion of the reaction, ice water was slowly added and the resulting solid was filtered to obtain compound 90-1 with a yield of 82% (10.1 g).

90-2. Synthesis of 5-bromo-1H-pyrrolo[2,3-b]pyridin-3-amine (Compound 90-2)

Compound 90-1 (5-bromo-3-nitro-1H-pyrrolo[2,3-b]pyridine; 10 g, 41.4 mmol), acetic acid (165 mL), Tin(II) chloride (23.5 g, 124.2 mmol), and hydrochloric acid (21 mL) were added and stirred at room temperature for 12 h. After the reaction was completed, ice water was slowly added, the pH was adjusted to 9, and the mixture was extracted with DCM. The organic layer was dried over MgSO ₄ and concentrated under reduced pressure to obtain compound 90-2 in a yield of 46% (4.1 g).

90-3. Synthesis of 4-(5-bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)morpholine (compound 90-3)

Compound 90-2 (5-bromo-1H-pyrrolo[2,3-b]pyridin-3-amine; 400 mg, 1.89 mmol), 2-bromoethyl ether (400 μL, 3.21 mmol), and N,N-diisopropylethylamine (670 μL, 3.87 mmol) were added to DMF (2 mL) and stirred at 110 °C for 2 h. After completion of the reaction, the mixture was extracted with water and EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. The residue was then separated by silica gel column chromatography to obtain compound 90-3 in a yield of 50% (266 mg).

90-4. Synthesis of 4-methoxy-3-(3-morpholino-1H-pyrrolo[2,3-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide (Compound 90)

3-Bromo-4-methoxy-N-(m-tolyl)benzenesulfonamide (50 mg, 0.14 mmol), bis(pinacolato)diboron (43 mg, 0.17 mmol), [1,1′-Bis(diphenylphosphino)ferrocene]dichloropalladium(II) (10 mg, 0.014 mmol), potassium acetate (41 mg, 0.42 mmol), and 1,4-dioxane (2 mL) were added and reacted using a microwave reactor at 120 ℃ for 1 h. After cooling to room temperature, compound 90-3 (4-(5-bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)morpholine; 39 mg, 0.14 mmol) and sodium carbonate aqueous solution (2 M, 0.15 mL) were added and reacted at 120 ℃ for 1 h. After the reaction was completed, the solvent was removed by celite filtration, extracted with water and EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. After separation by silica gel column chromatography, compound 90 was obtained with a yield of 11% (8 mg).

^1H NMR (300 MHz, DMSO-d ₆ ) δ 11.29 - 11.22 (m, 1H), 10.08 (s, 1H), 8.17 (d, J = 2.0 Hz, 1H), 7.93 (d, J = 2.1 Hz, 1H), 7.74 (dd, J = 8.6, 2.4 Hz, 1H), 7.68 (d, J = 2.4 Hz, 1H), 7.25 (d, J = 8.8 Hz, 1H), 7.13 (t, J = 8.0 Hz, 1H), 7.01 (d, J = 2.5 Hz, 1H), 6.95 (d, J = 6.9 Hz, 2H), 6.86 (d, J = 7.5 Hz, 1H), 3.82 (s, 3H), 3.78 (d, J = 4.8 Hz, 4H), 2.99 (t, J = 4.6 Hz, 4H), 2.21 (s, 3H).

Compound 91. 4-Methoxy-3-(3-morpholino-1H-pyrrolo[2,3-b]pyridin-5-yl)-N-(o-tolyl)benzenesulfonamide

Compound 91 (yield 37%) was synthesized using the same method as the synthesis of compound 90.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 11.25 (s, 1H), 9.43 (s, 1H), 8.16 (d, J = 2.1 Hz, 1H), 7.88 (d, J = 2.1 Hz, 1H), 7.66 (dd, J = 8.7, 2.4 Hz, 1H), 7.51 (d, J = 2.4 Hz, 1H), 7.27 (d, J = 8.7 Hz, 1H), 7.14 (dt, J = 9.5, 3.7 Hz, 3H), 7.02 (dd, J = 10.7, 3.2 Hz, 2H), 3.85 (s, 3H), 3.80 (t, J = 4.6 Hz, 4H), 2.98 (t, J = 4.6 Hz, 4H), 2.05 (s, 3H).

Compound 92. 4-Methoxy-3-(3-morpholino-1H-pyrrolo[2,3-b]pyridin-5-yl)-N-phenylbenzenesulfonamide

Compound 92 was synthesized using the same method as the synthesis of compound 90.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 8.49 (s, 1H), 8.09 (s, 1H), 7.86-7.74 (m, 2H), 7.69 (s, 1H), 7.26 (s, 1H), 7.22-7.11 (m, 3H), 7.00 (d, J = 8.5 Hz, 1H), 3.94 (q, J = 5.0 Hz, 4H), 3.87 (s, 3H), 3.56-3.42 (m, 4H).

Compound 93. N-(2-fluorophenyl)-4-methoxy-3-(3-morpholino-1H-pyrrolo[2,3-b]pyridin-5-yl)benzenesulfonamide

Compound 93 (yield 36%) was synthesized using the same method as the synthesis of compound 90.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 11.26 (s, 1H), 10.09 (s, 1H), 8.17 (d, J = 2.0 Hz, 1H), 7.93 (d, J = 2.0 Hz, 1H), 7.69 (dd, J = 8.6, 2.4 Hz, 1H), 7.63 (d, J = 2.4 Hz, 1H), 7.25 (d, J = 8.8 Hz, 1H), 7.16 - 7.09 (m, 4H), 7.00 (d, J = 2.5 Hz, 1H), 3.83 (s, 3H), 3.78 (d, J = 5.1 Hz, 4H), 2.99 (t, J = 4.7 Hz, 4H).

Compound 94. N-(2-chlorophenyl)-4-methoxy-3-(3-morpholino-1H-pyrrolo[2,3-b]pyridin-5-yl)benzenesulfonamide

Compound 94 (yield 36%) was synthesized using the same method as the synthesis of compound 90.

^1H NMR (300 MHz, DMSO-d ₆ ) δ 11.25 (s, 1H), 9.88 (s, 1H), 8.19 (d, J = 2.0 Hz, 1H), 7.91 (d, J = 2.1 Hz, 1H), 7.71 (dd, J = 8.7, 2.4 Hz, 1H), 7.63 (d, J = 2.4 Hz, 1H), 7.46 - 7.40 (m, 1H), 7.36 - 7.16 (m, 4H), 7.00 (d, J = 2.5 Hz, 1H), 3.85 (s, 3H), 3.79 (d, J = 4.8 Hz, 4H), 2.98 (t, J = 4.7 Hz, 4H).

Compound 95. N-(5-(2-methoxy-5-(N-phenylsulfamoyl)phenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)acetamide

95-1. Synthesis of N-(5-bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)acetamide (Compound 95-1)

5-Bromo-1H-pyrrolo[2,3-b]pyridin-3-amine (509 mg, 2.4 mmol) and THF (10 mL) were stirred at 0 °C, then trimethylamine (1 mL, 7.2 mmol) and acetic anhydride (450 μL, 4.8 mmol) were slowly added, and the mixture was stirred at room temperature for 5 h. After completion of the reaction, the mixture was extracted with water and EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain compound 95-1 in a yield of 60% (365 mg).

95-2. N-(5-(2-methoxy-5-(N-phenylsulfamoyl)phenyl)-1H-pyrrolo[2,3-b]

Synthesis of pyridin-3-yl)acetamide (compound 95)

3-Bromo-4-methoxy-N-phenylbenzenesulfonamide (89 mg, 0.26 mmol), bis(pinacolato)diboron (80 mg, 0.31 mmol), [1,1′-Bis(diphenylphosphino)ferrocene]dichloropalladium(II) (19 mg, 0.026 mmol), potassium acetate (76 mg, 0.78 mmol), and 1,4-dioxane (3 mL) were added and reacted at 120 ℃ for 1 h using a microwave reactor. After cooling to room temperature, compound 95-1 (N-(5-bromo-1H-pyrrolo[2,3-b]pyridin-3-yl)acetamide; 74 mg, 0.26 mmol) and sodium carbonate aqueous solution (2 M, 0.5 mL) were added and reacted at 120 ℃ for 1 h. After the reaction was completed, the solvent was removed by celite filtration, extracted with water and EtOAc, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. After separation by silica gel column chromatography, compound 95 was obtained with a yield of 11% (13 mg).

^1H NMR (300 MHz, Methanol-d ₄ ) δ 8.23 (d, J = 2.0 Hz, 1H), 8.18 (d, J = 2.1 Hz, 1H), 7.81 - 7.75 (m, 2H), 7.70 (d, J = 2.5 Hz, 1H), 7.31 - 7.06 (m, 7H), 3.90 (s, 3H), 2.22 (s, 3H).

Compound 96. N-(5-(2,4-difluoro-5-(N-(3-fluorophenyl)sulfamoyl)phenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)acetamide

Step A) Synthesis of 5-bromo-3-nitro-1H-pyrrolo[2,3-b]pyridine (Compound 96-1)

Nitric acid (6 mL) was slowly added to 5-bromo-1H-pyrrolo[2,3-b]pyridine (2.0 g, 10.15 mmol) at 0 ℃ and stirred for 1 hour. After completion of the reaction, the reactant was slowly added to ice water and the resulting solid was filtered to obtain compound 96-1 with a yield of 84% (2.06 g).

Step B) Synthesis of 5-bromo-1H-pyrrolo[2,3-b]pyridin-3-amine (compound 96-2)

5-Bromo-3-nitro-1H-pyrrolo[2,3-b]pyridine (750 mg, 3.1 mmol), Tin(II) chloride (2.94 g, 15.5 mmol), conc.HCl (62 mL), and ethanol (180 mL) were stirred at 100 °C for 2 h. After the reaction was completed, water was added, pH was adjusted to 10 with 1 N NaOH, and the mixture was extracted with DCM. The organic layer was dried over MgSO ₄ and concentrated under reduced pressure. The next reaction was carried out without purification.

Step C) Synthesis of N-(5-(2,4-difluoro-5-(N-(3-fluorophenyl)sulfamoyl)phenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)acetamide (Compound 96)

Compound 96-3 was synthesized using the same method as in Step A of the above compound 82. Then, compound 96 (yield 7%) was synthesized using the same method as in Step B of the above compound 26.

^1H NMR (400 MHz, DMSO _- d6 ) δ 11.58 (d, J = 2.6 Hz, 1H), 11.03 (s, 1H), 10.08 (s, 1H), 8.37 (s, 1H), 8.35 (d, J = 2.0 Hz, 1H), 8.02 (t, J = 8.1 Hz, 1H), 7.83 (d, J = 2.5 Hz, 1H), 7.73 (t, J = 10.2 Hz, 1H), 7.31 (td, J = 8.2, 6.7 Hz, 1H), 7.03 - 6.93 (m, 2H), 6.89 (td, J = 8.5, 2.6 Hz, 1H), 2.10 (s, 3H).

Compound 97. 3-(1H-Indol-5-yl)-4-methoxy-N-(m-tolyl)benzenesulfonamide

Compound 97 (yield 36%) was synthesized using the same method as the step B synthesis of compound 26 above.

^1H NMR (300 MHz, DMSO _-d6 ) δ 11.17 (s, 1H), 10.04 (s, 1H), 7.68 (dd, J = 8.6, 2.5 Hz, 1H), 7.63 (d, J = 2.4 Hz, 1H), 7.53-7.50 (m, 1H), 7.45-7.35 (m, 2H), 7.20 (d, J = 8.7 Hz, 1H), 7.17-7.08 (m, 2H), 6.98-6.90 (m, 2H), 6.89-6.83 (m, 1H), 6.46 (ddd, J = 3.0, 1.9, 0.9 Hz, 1H), 3.81 (s, 3H), 2.22 (s, 3H).

Compound 98. N-(3-fluorophenyl)-3-(1H-indol-5-yl)-4-methoxybenzenesulfonamide

Compound 98 (yield 37%) was synthesized using the same method as the step B synthesis of compound 26 above.

^1H NMR (300 MHz, DMSO _-d6 ) δ 11.18 (s, 1H), 10.42 (s, 1H), 7.72 (dd, J = 8.6, 2.5 Hz, 1H), 7.67 (d, J = 2.5 Hz, 1H), 7.55 (d, J = 1.6 Hz, 1H), 7.43 (dd, J = 8.4, 0.8 Hz, 1H), 7.38 (t, J = 2.7 Hz, 1H), 7.34-7.27 (m, 1H), 7.23 (d, J = 8.7 Hz, 1H), 7.13 (dd, J = 8.4, 1.7 Hz, 1H), 6.99-6.91 (m, 2H), 6.87 (tdd, J = 8.5, 2.5, 1.0 Hz, 1H), 6.47 (ddd, J = 3.0, 1.9, 0.9 Hz, 1H), 3.81 (s, 3H).

Compound 99. 4-Methoxy-N-(m-tolyl)-3-(3-((2-(trimethylsilyl)ethoxy)methyl)-3H-imidazo[4,5-b]pyridin-6-yl)benzenesulfonamide

Step A) Synthesis of 6-bromo-3-((2-(trimethylsilyl)ethoxy)methyl)-3H-imidazo[4,5-b]pyridine (Compound 99-1)

6-Bromo-1H-imidazo[4,5-b]pyridine (693 mg, 3.5 mmol) and DMF (35 mL) were stirred at 0 °C, then NaH (235 mg, 7.0 mmol) was added and stirred for 10 minutes. 2-(trimethylsilyl)ethoxymethyl chloride (990 μL, 5.95 mmol) was added and stirred at room temperature for 6 hours. After completion of the reaction, the mixture was quenched with water, extracted with DCM, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. The compound was then separated by silica gel column chromatography in a yield of 52% (597 mg).

Step B) Synthesis of 4-methoxy-N-(m-tolyl)-3-(3-((2-(trimethylsilyl)ethoxy)methyl)-3H-imidazo[4,5-b]pyridin-6-yl)benzenesulfonamide (Compound 99)

Compound 99 (yield 36%) was synthesized using the same method as the step B synthesis of compound 26 above.

^1H NMR (300 MHz, DMSO _-d6 ) δ 10.06 (s, 1H), 8.67 (s, 1H), 8.37 (d, J = 1.9 Hz, 1H), 8.10 (d, J = 2.0 Hz, 1H), 7.77 (dd, J = 8.7, 2.4 Hz, 1H), 7.70 (d, J = 2.4 Hz, 1H), 7.28 (d, J = 8.7 Hz, 1H), 7.13 (t, J = 7.6 Hz, 1H), 6.99 - 6.92 (m, 2H), 6.86 (d, J = 7.5 Hz, 1H), 5.68 (s, 2H), 3.84 (s, 3H), 3.69-3.56 (m, 2H), 2.21 (s, 3H), 0.86 (t, J = 8.0 Hz, 2H), -0.07 (s, 9H).

Compound 100. 3-(3H-imidazo[4,5-b]pyridin-6-yl)-4-methoxy-N-(m-tolyl)benzenesulfonamide

4-methoxy-N-(m-tolyl)-3-(3-((2-(trimethylsilyl)ethoxy)methyl)-3H-imidazo[4,5-b]pyridin-6-yl)benzenesulfonamide (100 mg, 0.19 mmol) and THF (2 mL) were mixed with TBAF (500 mg, 1.91 mmol) and refluxed for 24 h. After completion of the reaction, the mixture was extracted with water and DCM, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain compound 100 in a yield of 42% (32 mg).

^1H NMR (300 MHz, DMSO _-d6 ) δ 13.06 (s, 1H), 10.06 (s, 1H), 8.50 (s, 1H), 8.33 (d, J = 2.0 Hz, 1H), 8.02 (d, J = 2.0 Hz, 1H), 7.76 (dd, J = 8.7, 2.5 Hz, 1H), 7.71 (d, J = 2.4 Hz, 1H), 7.27 (d, J = 8.7 Hz, 1H), 7.13 (t, J = 7.7 Hz, 1H), 6.98 - 6.91 (m, 2H), 6.86 (d, J = 7.6 Hz, 1H), 3.84 (s, 3H), 2.22 (s, 3H).

Compound 101. N-(3-fluorophenyl)-4-methoxy-3-(3-((2-(trimethylsilyl)ethoxy)methyl)-3H-imidazo[4,5-b]pyridin-6-yl)benzenesulfonamide

Compound 101 (yield 27%) was synthesized using the same method as the synthesis of compound 99.

^1H NMR (300 MHz, DMSO _-d6 ) δ 10.44 (s, 1H), 8.67 (s, 1H), 8.39 (d, J = 1.9 Hz, 1H), 8.14 (d, J = 2.0 Hz, 1H), 7.81 (dd, J = 8.7, 2.4 Hz, 1H), 7.75 (d, J = 2.4 Hz, 1H), 7.35-7.25 (m, 2H), 7.02-6.92 (m, 2H), 6.92-6.83 (m, 1H), 5.68 (s, 2H), 3.84 (s, 3H), 3.62 (t, J = 8.1 Hz, 2H), 0.86 (t, J = 8.0 Hz, 2H), -0.08 (s, 9H).

Compound 102. N-(3-fluorophenyl)-3-(3H-imidazo[4,5-b]pyridin-6-yl)-4-methoxybenzenesulfonamide

Compound 102 (yield 13%) was synthesized using the same method as the synthesis of compound 100.

^1H NMR (300 MHz, DMSO _-d6 ) δ 13.22 (s, 1H), 12.79 (s, 1H), 10.45 (s, 1H), 8.50 (s, 1H), 8.35 (s, 1H), 8.05 (s, 1H), 7.80 (dd, J = 8.5, 2.5 Hz, 1H), 7.75 (d, J = 2.5 Hz, 1H), 7.30 (q, J = 7.5 Hz, 2H), 7.03-6.92 (m, 2H), 6.87 (t, J = 8.6 Hz, 1H), 3.84 (s, 3H).

Compound 103. 4-Methoxy-3-(4-oxo-1,4-dihydroquinolin-6-yl)-N-(m-tolyl)benzenesulfonamide

Compound 103 (yield 11%) was synthesized using the same method as the step B synthesis method of compound 26 above.

^1H NMR (300 MHz, DMSO- d ₆ ) δ 11.85 (d, J = 5.7 Hz, 1H), 10.12 (s, 1H), 8.11 (d, J = 2.1 Hz, 1H), 7.93 (dd, J = 7.4, 5.8 Hz, 1H), 7.75 - 7.69 (m, 3H), 7.58 (d, J = 8.6 Hz, 1H), 7.25 (d, J = 9.4 Hz, 1H), 7.12 (t, J = 7.7 Hz, 1H), 6.93 (d, J = 9.3 Hz, 2H), 6.85 (d, J = 7.5 Hz, 1H), 6.07 (d, J = 7.3 Hz, 1H), 3.83 (s, 3H), 2.21 (s, 3H).

Compound 104. N-(3-fluorophenyl)-4-methoxy-3-(4-oxo-1,4-dihydroquinolin-6-yl)benzenesulfonamide

Compound 104 (yield 10%) was synthesized using the same method as the step B synthesis of compound 26 above.

^1H NMR (300 MHz, DMSO _-d6 ) δ 11.86 (d, J = 5.7 Hz, 1H), 10.50 (s, 1H), 8.12 (d, J = 2.1 Hz, 1H), 7.93 (dd, J = 7.4, 5.7 Hz, 1H), 7.79-7.72 (m, 3H), 7.59 (d, J = 8.7 Hz, 1H), 7.34-7.24 (m, 2H), 6.99-6.81 (m, 3H), 6.07 (d, J = 7.4 Hz, 1H), 3.84 (s, 3H).

Compound 105. 3-(4-oxo-1,4-dihydroquinolin-6-yl)-N-(m-tolyl)benzenesulfonamide

Compound 105 (yield 20%) was synthesized using the same method as the step B synthesis of compound 26 above.

^1H NMR (400 MHz, DMSO- d ₆ ) δ 11.91 (s, 1H), 10.31 (s, 1H), 8.38 - 8.32 (m, 1H), 8.11 (s, 1H), 8.00-7.91 (m, 3H), 7.72 (d, J = 7.8 Hz, 1H), 7.69-7.62 (m, 2H), 7.10 (t, J = 7.7 Hz, 1H), 6.98-6.87 (m, 2H), 6.84 (d, J = 7.6 Hz, 1H), 6.10 (d, J = 7.5 Hz, 1H), 2.19 (s, 3H).

Experimental Example 1. Confirmation of the Influence of IRP2 Protein on Cancer Cell Proliferation and Iron Homeostasis

Cancer cells are known to require relatively higher iron content than normal cells for cell division, proliferation, and metastasis (Heath et al., 2013). To determine the effect of iron on cancer cell growth, we initially supplemented intracellular iron by treating cells with ferric ammonium citrate (FAC, Fe ³⁺ , (NH ₄ ) ₅ [Fe(C ₆ H ₄ O ₇ ) ₂ ]) (Plath et al., 2015). FAC treatment increased the growth of colon cancer cell lines (DLD-1, HCT116, HCT15, LOVO, SW480, SW620) in a time-dependent manner, whereas treatment with deferoxamine (DFO), which induces iron depletion, significantly inhibited cell growth (Figure 8A). Therefore, we confirmed that the amount of intracellular iron is an important factor that can determine the proliferation rate of cancer cells. Next, we investigated whether FAC or DFO treatment induces changes in iron metabolism-related factors. Previous publications have highlighted the importance of iron regulatory proteins (IRPs) in maintaining iron homeostasis in cancer cells (Khiroya et al., 2017). Therefore, we examined the expression of IRP proteins by FAC and DFO treatment. FAC decreased IRP2 expression, whereas DFO treatment increased IRP2 expression and significantly altered the expression of its downstream proteins, transferrin receptor 1 (TfR1) and ferritin H (FTH). IRP1 expression was unaffected (Fig. 8B).

Because IRP2, rather than IRP1, is specifically implicated in iron metabolism dysregulation in colorectal cancer, we investigated the effect of IRP2 on cancer cell growth. Using small interfering RNA (siRNA) and clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 to specifically target IRP2 for deletion, we observed a significant inhibition of viability of IRP2-deleted cells compared to control cells (Figures 1A and 1B). Furthermore, we evaluated the tumorigenicity of IRP2 knockout (KO) SW480 cells in vivo. We demonstrated that tumorigenicity of IRP2 KO cells was significantly delayed compared to that of WT cells (Figures 1C, 1D, and 1E). Overall, these results indicate that IRP2 is crucial for determining cell proliferation through the regulation of iron homeostasis.

Additionally, we compared the patterns of the top 100 genes with positive or negative correlations with IRP2 expression using the Cancer Genome Atlas (TCGA) database and analyzed them as a heatmap (Fig. 1F). Gene ontology analysis confirmed a positive correlation between IRP2 expression and genes related to ubiquitination-related signaling pathways and cell differentiation (Fig. 1G). Additionally, using a Kaplan-Meier curve to compare survival outcomes according to IRP2 expression, we confirmed that patients with IRP2 overexpression had a poor survival prognosis (Fig. 1H), emphasizing the importance of IRP2 as a therapeutic target for colon cancer.

The present inventors also compared IRP1/2 mRNA and protein expression in paired normal and tumor tissues derived from colorectal cancer patients. Interestingly, IRP2 mRNA and protein expression was overexpressed by more than 1.5-fold in 5 out of 10 colon patient tissues, whereas IRP1 expression did not differ significantly between normal and tumor tissues (Figures 9A and 9B). Therefore, we confirmed that IRP2, rather than IRP1, is a more important factor in colorectal cancer cell growth.

<Experimental Example 2. Identification of a Novel Small Molecule that Interferes with IRP2 Binding to IRE>

We aimed to identify novel small molecules that interfere with IRP2 binding to IRE (Fig. 2A). Homology modeling was performed to construct the three-dimensional structure of the IRP2-IRE complex based on the X-ray crystal structure of the IRP1/IRE complex (PDB ID: 3SNP) and the cryo-EM structure of IRP2 (PDB ID: 6VCD) (Selezneva et al., 2013; Wang et al., 2020). Furthermore, because the sequence identity between IRP1 and IRP2 is greater than 60%, mutational data from IRP1 could be used to identify key target residues of IRP2 (Selezneva et al., 2013). The present inventors found that S446 of IRP2 interacts with N7 of adenine (A15) in IRE through a hydrogen bond, R454 interacts with N7 of guanine (G16) in IRE through a hydrogen bond, and hydrophobic interactions are formed between A15 and G16 of IRE and I333, S444, V445, S446, P451, R454, and N610 of IRP2 (Fig. 2B).

To construct the pharmacophore model, we considered not only residues that directly hydrogen-bond with IRE but also residues that constitute the hydrophobic pocket where the IRE terminal loop binds (Fig. 2C). In the generated pharmacophore model, D334, S446, R454, and N610 were selected as key residues, and overall, the final structure-based pharmacophore model consists of seven pharmacophore functions: one hydrogen bond donor (HBD), three hydrogen bond acceptors (HBAs), one hydrophobic (Hy), and two cyclic aromatic (RA) functions. The three HBA functions were derived from the hydroxyl group of S446, the quinidine group of Arg454, and the backbone of N610, respectively. Two RA functions were added considering the purine base characteristics of A15 and G16 of IRE (Figs. 2B and 2C). One hydrophobic function reflected the hydrophobic terminal loop binding pocket generated by D334 and one HBD function (Fig. 2C).

Based on the pharmacophore model generated by the inventors, a virtual screening was performed using an in-house database (DB) containing 8 million compounds. A primary screening resulted in 5,782 compounds, which were then filtered for a fitness score of 1.0 or higher and further visualized for selection. Finally, 32 compounds were screened as virtual hits based on the pharmacophore model (Figure 10A), and among these compounds, Hit02 exhibited potent cytotoxicity in colon cancer cell lines (Figure 10B). Subsequently, chemical modifications were performed based on the X-ray crystal structure of Hit02. Compounds 1 and 44 exhibited excellent antitumor activity and improved solubility, and binding to IRP2 was demonstrated by surface plasmon resonance (SPR) analysis using K _D (Figures 2D, 2E, and 11). Furthermore, RNA-protein pull-down assays were performed to evaluate the binding ability of IRP2/IRE. As expected, compounds 1 and 44 altered the IRP/IRE system, which was also confirmed by similar results in RNA-immunoprecipitation, whereby IRP2 binding to FTH mRNA was reduced (Figures 2F and 2G).

Molecular docking studies were performed using Glide XP docking to predict the binding modes of compounds 1 and 44. Compound 1 appears to be stabilized by hydrogen bonding interactions with Asp334, Ser444, Arg454, and Thr513, π-cation interactions with Arg454, and hydrophobic interactions with Ile333, Leu337, Ile441, Cys512, and Leu626 ( Figure 2H ). Compound 44 exhibits hydrogen bonding interactions with Ser444, Arg454, and Asn514, π-cation interactions with Arg454, and hydrophobic interactions with Ile333 and Cys523 ( Figure 2I ). Therefore, compounds 1 and 44 have selectivity for IRP2 binding to IRE and are suitable class 1 inhibitors for reducing the IRP2/IRE system in cells.

Experimental Example 3. Cancer cell toxicity through iron metabolism reprogramming induced by compounds 1 and 44.

The present inventors assessed the antitumor effects of compounds 1 and 44 by measuring cell viability in colon cancer cells. These compounds exhibited significant cytotoxicity in an IRP2-dependent manner and did not affect proliferation in cells lacking IRP2 (Fig. 3A). In contrast, they exhibited slight cytotoxicity at high doses in various normal cell lines, including CCD-18Co, VERO, HFL-1, L929, NIH 3T3, and CHO-K1 (Fig. 3A).

Since spheroids, a three-dimensional (3D) culture model, can more accurately reflect patient tumor tissues than 2D monolayer cell cultures (Song et al., 2018), the cancer cell growth inhibitory effects of compounds 1 and 44 were evaluated in 3D spheroid models of SW480 and LOVO cell lines. These compounds exhibited antiproliferative activity as evidenced by an increase in ethidium homodimer-1 (EthD-1) intensity, which was indicative of a decrease in spheroid area and an increase in the number of dead cells ( Figure 3B ). In addition, we performed cell cycle analysis to quantify the percentage of cells in different phases. Compounds 1 and 44 exhibited a significant accumulation of cells in the G2/M phase along with a decrease in the G0/G1 phase ( Figure 12A ).

We investigated whether compounds 1 and 44 induced changes in iron metabolism by targeting IRP2 binding to IRE. As expected, these compounds decreased the expression of TfR1, which is responsible for iron uptake in cells, and increased the translational activation of FTH, a key component of iron storage, by reducing IRP2 (Fig. 3C), but did not affect the transcriptional activity of IRP2 (Fig. 12B).

Next, we observed the stability of IRP2 and its downstream proteins when treated with cycloheximide (CHX), a protein translation inhibitor. When treated simultaneously with CHX and compound 1, compound 1 did not affect the translational activity of proteins such as IRP1, IRP2, and TfR1, but rather exhibited the structure of the FTH protein within 24 hours (Figure 12C). These results demonstrated that compound 1 sensitively affected FTH translational activity and further modulated the translation of downstream proteins by inhibiting IRP2 protein expression. Furthermore, immunostaining revealed that compounds 1 and compound 44 reduced cytoskeletal formation and IRP2 protein expression compared to vehicle treatment (Figure 3D). Collectively, these results suggest that compounds 1 and compound 44 exert cytotoxic effects through reprogramming iron metabolism.

<Experimental Example 4. Confirmation of the effects of compounds 1 and 44 on reducing intracellular iron deficiency and ROS through ubiquitin-dependent degradation of IRP2>

To elucidate the cause of the decrease in IRP2 protein expression after treatment with Compound 1, ubiquitination of the IRP2 protein was analyzed. As shown in Figure 4A, Compound 1 induced ubiquitination of the IRP2 protein similar to treatment with MG-132, a proteasome inhibitor used as a positive control. Therefore, Compound 1 was confirmed to reduce IRP2 protein through a ubiquitin-dependent mechanism.

Since the reduction of TfR1 induced by IRP2 inhibition affects intracellular iron uptake, we hypothesized that intracellular free iron (LIP) would be reduced through the degradation of IRP2 protein by compounds 1 and 44. The fluorescence of calcein acetoxymethyl (Calcein)-ester (AM), a LIP measurement probe, decreases after chelation of iron, and the amount of iron can be estimated by the degree of fluorescence decrease. The intensity of calcein-AM was increased by treatment with compounds 1 and 44 and with di-2-puridylketone-4-cyclohexyl-4-methyl-3-thiosemicarbazone (DPC), an iron chelator that forms a complex bond with iron to reduce free iron (Figs. 4B and 13D). Since a link between iron and reactive oxygen species (ROS) has been reported (Nakamura et al., 2019), intracellular ROS was measured by staining with 2',7'-Dicholoroflurescin diacetate (DCFDA), a cellular ROS marker. Compounds 1 and 44 substantially reduced ROS production by scavenging free iron from cells ( Figure 4C ). Collectively, these results indicate that ubiquitin-dependent degradation of IRP2 induced by compounds 1 and 44 disrupts the maintenance of iron homeostasis after LIP and ROS reduction.

Iron is utilized in various mitochondrial functions such as ATP production, iron-sulfur cluster biogenesis, and respiration (Bauckman et al., 2015), and intracellular iron deficiency has been reported to impair mitochondrial activity due to the deficiency of iron-sulfur complexes (Cloonan et al., 2016; Li et al., 2019). Therefore, we predicted that IRP2 inhibitors would induce iron deficiency through changes in iron metabolism and thus affect mitochondrial function. Compound 1 inhibited mitochondrial oxidative phosphorylation (OXPHOS), reducing oxygen consumption rate (OCR), basal respiration, and ATP production (Figures 4D and 13A), whereas it activated glycolysis, showing an increase in basal and compensatory glycolysis (Figures 4D and 13B).

<Experimental Example 5. Gene Expression Profiling of Colon Cancer Cells Treated with IRP2 Inhibitors>

To investigate the overall effects of IRP2 inhibitors in colon cancer cells, we additionally performed gene set enrichment analysis (GSEA) on RNA-sequencing data. Consistent with previous results showing that compound 1 exerts antiproliferative effects through cell cycle arrest by significantly suppressing genes involved in E2F targets, the G2M checkpoint, and mitosis (Figure 5A). Compound 1 upregulated the expression of genes involved in hypoxia, epithelial-mesenchymal transition, angiogenesis, and glycolysis, while suppressing the expression of genes involved in mitochondrial oxidative phosphorylation (OXPHOS) (Figures 5A and 5B). Inhibition of IRP2 binding to IRE in the 5'-UTR of mRNA upregulated the translation of hypoxia-inducible factor (HIF), which transcriptionally activates the expression of target genes involved in glucose metabolism, angiogenesis, invasion, and metastasis (Hong S-S et al., 2004). Furthermore, since HIF-mediated hypoxia upregulation is associated with metabolic changes, compound 1 exhibits metabolic reprogramming from OXPHOS to glycolysis (Figures 4D, 5A, and 5B). The enhancement of the HIF pathway by compound 1 suggests a compensatory mechanism for cancer cell survival by IRP2 inhibition. In particular, compound 1 increased pathways related to the unfolded protein response and autophagy, while decreasing mTORC1 signaling. Mitochondrial dysfunction leads to mitophagy, which is the selective degradation of mitochondria by autophagy (Youn DH et al., 2021), and stimulates AMP-activated protein kinase (AMPK), which activates Unc-51-like autophagy-activating kinase-1 (ULK1) and Beclin-1, which acts as an autophagy promoter (You L et al., 2015). This suggests that disruption of iron metabolism targeting IRP2 leads to activation of autophagy via mitochondrial dysfunction.

<Experimental Example 6. Confirmation of the promotion of autophagy-induced cancer cell death through the AMPK-ULK1-Beclin1-LC3B pathway induced by compounds 1 and 44>

After confirming the activation of autophagy target genes by compound 1, we next examined whether autophagic death was indeed induced by compound 1 and compound 44. Genetic deletion of IRP2 in SW480 cells resulted in the formation of cellular multivesicles and autophagosomes, as observed using transmission electron microscopy (TEM) (Fig. 6A), and pharmacological inhibition of IRP2 by compound 1 and compound 44 showed similar results (Fig. 6B). Furthermore, the expression of MPK, ULK1, Beclin-1, and LC3B proteins was significantly increased in IRP2-deleted SW480 cells compared to WT cells, whereas the addition of IRP2 suppressed the expression of these proteins (Fig. 6C). Pharmacological inhibition of IRP2 by compound 1 and compound 44 increased AMPK phosphorylation and sequentially increased the expression of ULK1 and Beclin-1 over time (Fig. 6D). Finally, we re-examined the expression of LC3B, an important marker of autophagic death, using immunostaining, and found that the expression of LC3B was significantly increased by compounds 1 and 44, as shown in Figure 6E.

Next, to assess whether inhibition of IRP2 is dependent on autophagy, we confirmed the expression of B-cell lymphoma 2 (Bcl2), cleaved caspase-3 (regulator of the apoptosis pathway), and glutathione peroxidase-4 (GPX4) (regulator of iron-dependent apoptosis) in IRP2-deficient SW480 cells. The expression of these proteins was not altered by genetic deletion of IRP2 in SW480 cells, indicating that inhibition of IRP2 induces autophagy-dependent cell death, not through the apoptosis pathway and iron-dependent cell death (Fig. 13C). This suggests that the novel IRP2 inhibitor exhibits antitumor effects in colon cancer through an autophagy-dependent cell death mechanism that is dependent on activation of the AMPK-ULK1-Beclin1-LC3B pathway.

<Experimental Example 7. Confirmation of the various sensitivities of colon cancer organoids to compounds 1 and 44 and their in vivo tumor growth inhibition>

Patient-derived organoids have recently been explored as platforms for applications ranging from drug discovery to cancer therapy (Kim et al., 2019). Therefore, we treated nine established colorectal cancer organoids with compound 1 and monitored morphological changes and cell viability using CellTier-Glo. Organoids treated with compound 1 exhibited complete destruction of organoid structure as well as cell death, with differential sensitivity ranging from 0.5 to 40 μM IC (Figure _7A ). For further study, organoids were divided into the most and least sensitive to compound 1, and IRP2 protein expression was significantly reduced in the most sensitive organoid (Figures 7B and 7C), confirming a decrease in IRP2 protein expression after compound 1 treatment in colorectal cancer organoids. Notably, the expression level of IRP2 correlated with organoid sensitivity to compound 1, while no correlation was observed for IRP1 (Figure 7D).

Finally, the in vivo antitumor activity of compounds 1 and 44 was evaluated using an established xenograft mouse model injected subcutaneously into the SW480 cell line. Intraperitoneal administration of compounds 1 and 44 (100 mpk) significantly reduced tumor volume (Figures 7E and 7F) and effectively reduced tumor weight (Figure 7G). To further validate the intracellular results, IRP2 expression was examined in tumors. Compound 1 and 44 treatment groups showed a decrease in IRP2 expression compared to the vehicle treatment group (Figure 7H). Collectively, these results suggest that novel small-molecule inhibitors targeting IRP2 exhibit effective tumor suppression responses in vivo.

<Experimental Example 8. Anticancer Activity>

The anticancer activity of the compound of the present invention was confirmed by measuring the % or IC ₅₀ value using CCK.

8-1. Cell culture

Human colon cancer cell lines were purchased from the Korea Cell Line Bank. Cell lines were maintained in appropriate complete growth media, primarily Dulbecco's Modified Eagle Medium (DMEM) and Roswell Park Memorial Institute-1640 (RPMI-1640) supplemented with 10% fetal bovine serum (FBS), 100 U/ml penicillin, and 100 μg/ml streptomycin. Cell lines were cultured at 37°C in a 5% _CO2 incubator.

8-2. Cell survival analysis

Cell viability was measured using the Cell Counting Kit-8 (CCK-8). An appropriate number of cells (2 × 10 ⁴ ) were seeded in 96-well plates and cultured for 24 h. The cells were then treated with various concentrations of compounds 1 and 44 for 48 h. After adding CCK-8 solution and incubating for 3 h at 37°C, the absorbance was measured at 450 nm using a microplate reader, and the experiment was performed in triplicate.

compound CCK % HCT116
at 1μM SW480 IC ₅₀
(μM) LOVO IC ₅₀
(μM) Compound 1 36 1 5 Compound 2 7 2 Compound 3 49 18 Compound 5 28 8 Compound 6 29 2 Compound 7 2 0.5 Compound 8 1 30 Compound 9 0.1 19 Compound 10 2 0.8 Compound 11 1 11 Compound 12 1 5 Compound 14 61 　 　 Compound 18 63 N/A Compound 19 59 56 Compound 20 15 31 Compound 21 3 11 Compound 23 68 N/A Compound 24 0.4 4 Compound 25 1 4 Compound 26 62 72 Compound 27 >100 >100 Compound 28 1 11 Compound 29 2 7 Compound 30 0.5 5 Compound 31 0.5 3 Compound 33 2 6 Compound 34 N/A N/A Compound 35 31 23 Compound 36 4 3 Compound 37 2 3 Compound 38 7 21 Compound 39 0.03 0.4 Compound 40 15 10 Compound 43 32 　 　 Compound 44 0.2 4 Compound 45 7 3 Compound 46 2 3 Compound 47 2 11 Compound 48 0.002 1 Compound 49 14 19 Compound 57 85 Compound 58 90 Compound 59 85 Compound 60 2 16 Compound 61 2 17 Compound 62 0.8 5 Compound 63 0.4 2 Compound 64 32 47 Compound 67 30 Compound 68 0.5 12 Compound 71 65 Compound 72 99 Compound 74 N/A 60 Compound 77 8 16 Compound 78 94 Compound 79 12 16 Compound 80 4 2 Compound 81 0.1 0.2 Compound 82 18 14 Compound 83 5 4 Compound 84 16 14 Compound 85 2 1 Compound 86 79 Compound 87 4 6 Compound 88 0.6 0.1 Compound 89 0.9 1 Compound 95 20 　 　 Compound 97 7 8 Compound 98 13 8 Compound 100 19 29 Compound 101 >100 13 Compound 102 5 2 Compound 103 32 82 Compound 104 33 61 Compound 105 7 16

Looking at the above Table 1, it was confirmed that compounds 1 to 105 of the present invention have a cell killing effect on cancer cells. In particular, in chemical formulas 2 to 5, which are benzenesulfonamide-substituted heterobicyclic derivatives, R ₁ is substituted with hydrogen, haloalkyl, alkyl, acetyl, acetamido, phenyl, thiophenyl or morpholinyl; R ₂ is substituted with hydrogen, alkyl or acetyl; R ₃ is substituted with hydrogen, halogen or C ₁ -C ₆ alkoxy; and R ₄ is or The compound substituted with showed excellent anticancer activity.

<Example 1. Preparation of a powder>

1.2g of the compound of the present invention and 1g of lactose were mixed and filled into a sealed bag to prepare a powder.

<Example 2. Preparation of tablets>

100 mg of the compound of the present invention 1, 100 mg of microcrystalline cellulose, 60 mg of lactose monohydrate, 20 mg of low-substituted hydroxypropyl cellulose, and 2 mg of magnesium stearate were mixed and then compressed into tablets according to a conventional tablet manufacturing method.

<Example 3. Preparation of capsules>

100 mg of the compound of the present invention 1, 100 mg of microcrystalline cellulose, 60 mg of lactose monohydrate, 20 mg of low-substituted hydroxypropyl cellulose, and 2 mg of magnesium stearate were mixed, and then the above ingredients were mixed according to a conventional capsule manufacturing method and filled into a gelatin capsule to manufacture a capsule.

<Example 4. Preparation of the pill>

90 mg of the compound of the present invention 1, 5 mg of glutinous rice starch, 5 mg of purified water, and a small amount of dextrin, maltodextrin, corn starch, and microcrystalline cellulose (MCC) as additives to inhibit hygroscopicity were mixed, and then 100 mg of pills were made according to a conventional method.

<Example 5. Preparation of injection>

After mixing 10 mg of the compound of the present invention, an appropriate amount of sterile distilled water for injection, and an appropriate amount of a pH regulator, the above-mentioned ingredient content per 1 ampoule (2 ml) was prepared according to a conventional method for preparing injections.

Claims (14)

A pharmaceutical composition for preventing or treating colon cancer, comprising a compound represented by the following chemical formula 1, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof as an active ingredient:
[Chemical Formula 1]

In the above chemical formula 1,
R ₁ is substituted with one or more substituents selected from the group consisting of hydrogen, halogen, hydroxy, amino, amide, cyano, halo C ₁ -C ₆ alkyl, NO ₂ , C _{1 -C 6} alkyl _, C ₁ -C ₁₀ alkoxy, acetyl, acetamido, C ₁ -C ₆ alkylamino, di(C ₁ -C ₆ ) alkylamino, C ₁ -C ₆ alkylcarbonylamino, C ₅ -C ₁₀ aryl, C ₅ -C ₁₀ heteroaryl, C 4 -C 10 cycloalkyl and C ₄ -C ₁₀ heterocycloalkyl;
R ₂ is hydrogen, C ₁ -C ₆ alkyl, halo C ₁ -C ₆ alkyl, C ₁ -C ₁₀ alkoxy, acetyl, C ₁ -C ₆ alkylamino, di(C ₁ -C ₆ )alkylamino, di(C ₁ -C ₆₎ alkylamino (C ₁ -C ₆ ) Substituted with one or more substituents selected from the group consisting of alkyl, phenyl, and SEM(=2-(trimethylsilyl)ethoxymethyl);
R ₃ is independently hydrogen, halogen or C ₁ -C ₁₀ alkoxy;
R ₄ is substituted with one or more substituents selected from the group consisting of substituted or unsubstituted (C ₅ -C ₁₀ )aryl-amino, substituted or unsubstituted (C ₅ -C ₁₀ )heteroaryl-amino, substituted or unsubstituted C ₅ -C ₁₀ heteroaryl, and substituted or unsubstituted C ₄ -C ₁₀ heterocycloalkyl,
The above substituted (C ₅ -C ₁₀ )aryl-amino, (C ₅ -C ₁₀ )heteroaryl-amino, C ₅ -C ₁₀ heteroaryl and C ₄ -C ₁₀ heterocycloalkyl are substituted with one or more substituents independently selected from the group consisting of hydrogen, halogen, hydroxy, amide, halo C ₁ -C ₆ alkyl, cyano, NO ₂ , C ₁ -C ₆ alkyl, acetyl, carbamoyl, substituted or unsubstituted C ₁ -C ₁₀ alkoxy, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, and substituted or unsubstituted C ₄ -C ₁₀ heterocycloalkyl,
The above substituted C ₁ -C ₁₀ alkoxy, C ₃ -C ₁₀ cycloalkyl or C ₄ -C ₁₀ heterocycloalkyl is substituted with one or more substituents selected from the group consisting of hydrogen, halogen, hydroxy, amide, halo C ₁ -C ₆ alkyl, cyano, NO ₂ , C ₁ -C ₆ alkyl, and C ₁ -C ₁₀ alkoxy;
A is CR ₁ , CH or N;
is a single bond or double bond;
n is an integer from 0 to 3; In the first paragraph,
In the above chemical formula 1,
R ₁ is hydrogen, halogen, hydroxy, amino, amide, CN, halo C ₁ -C ₆ alkyl, NO ₂ , C ₁ -C ₆ alkyl, C ₁ -C ₁₀ alkoxy, acetyl, acetamido, C ₁ -C ₆ alkylamino, di(C ₁ -C ₆ ) Substituted with one or more substituents selected from the group consisting of alkylamino, C ₁ -C ₆ alkylcarbonylamino, phenyl, thiophenyl, and morpholinyl;
R ₂ is substituted with one or more substituents selected from the group consisting of hydrogen, halogen, hydroxy, amide, CN, halo C ₁ -C ₆ alkyl, NO ₂ , C _{1 -C 6} alkyl, C ₁ -C ₁₀ alkoxy, acetyl, C ₁ -C ₆ alkylamino, di(C ₁ -C _{6 )} alkylamino, di(C ₁ -C _{6) alkylamino(C 1 -C 6)} alkyl, phenyl, and SEM(=2-(trimethylsilyl)ethoxymethyl);
R ₃ is independently hydrogen, halogen or C ₁ -C ₁₀ alkoxy;
R ₄ is substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted and substituted or unsubstituted is substituted with one or more substituents selected from the group consisting of,
The above substituted , , , , and is substituted with one or more substituents independently selected from the group consisting of hydrogen, halogen, hydroxy, amide, halo C ₁ -C ₆ alkyl, CN, NO ₂ , C ₁ -C ₆ alkyl, acetyl, carbamoyl, substituted or unsubstituted C ₁ -C ₁₀ alkoxy, substituted or unsubstituted C ₄ -C ₁₀ cycloalkyl, and substituted or unsubstituted C ₄ -C ₁₀ heterocycloalkyl,
The above substituted C ₁ -C ₁₀ alkoxy, C ₃ -C ₁₀ cycloalkyl or C ₄ -C ₁₀ heterocycloalkyl is substituted with one or more substituents selected from the group consisting of hydrogen, halogen, hydroxy, amide, halo C ₁ -C ₆ alkyl, CN, NO ₂ , C ₁ -C ₆ alkyl, and C ₁ -C ₁₀ alkoxy;
A is CR ₁ , CH or N;
is a single bond or double bond;
n is an integer from 0 to 3;
A pharmaceutical composition for preventing or treating colon cancer, characterized by: A pharmaceutical composition for preventing or treating colon cancer, comprising a compound represented by the following chemical formula 2, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof as an active ingredient:
[Chemical Formula 2]

In the above chemical formula 2,
R ₁ is substituted with one or more substituents selected from the _group consisting of hydrogen, halogen, hydroxy, amino, amide, CN, halo C ₁ -C ₆ alkyl, NO ₂ , C _{1 -C 6} alkyl, C ₁ -C ₁₀ alkoxy, acetyl, acetamido, C 1 -C 6 alkylamino, di(C ₁ -C ₆₎ alkylamino, C ₁ -C ₆ alkylcarbonylamino, phenyl, thiophenyl, and morpholinyl;
R ₂ is substituted with one or more substituents selected from the group consisting of hydrogen, halogen, hydroxy, amide, CN, halo C ₁ -C ₆ alkyl, NO ₂ , C _{1 -C 6} alkyl, C ₁ -C ₁₀ alkoxy, acetyl, C ₁ -C ₆ alkylamino, di(C ₁ -C _{6 )} alkylamino, di(C ₁ -C _{6) alkylamino (C 1 -C 6)} alkyl, phenyl, and SEM(=2-(trimethylsilyl)ethoxymethyl);
R ₃ is independently hydrogen, halogen or C ₁ -C ₁₀ alkoxy;
R ₄ is substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted and substituted or unsubstituted is substituted with one or more substituents selected from the group consisting of,
The above substituted , , , , and is substituted with one or more substituents independently selected from the group consisting of hydrogen, halogen, hydroxy, amide, halo C ₁ -C ₆ alkyl, CN, NO ₂ , C ₁ -C ₆ alkyl, acetyl, carbamoyl, substituted or unsubstituted C ₁ -C ₁₀ alkoxy, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted C ₄ -C ₁₀ heterocycloalkyl,
The above substituted C ₁ -C ₁₀ alkoxy, C ₃ -C ₁₀ cycloalkyl or C ₄ -C ₁₀ heterocycloalkyl is substituted with one or more substituents selected from the group consisting of hydrogen, halogen, hydroxy, amide, halo C1-C6 alkyl, CN, NO ₂ , C ₁ -C ₆ alkyl, C ₁ -C ₁₀ alkoxy;
n is an integer from 0 to 3; In the third paragraph,
In the above chemical formula 2,
R ₁ is substituted with one or more substituents selected from the group consisting of hydrogen, halo C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl, acetyl, acetamido, phenyl, thiophenyl, and morpholinyl;
R ₂ is substituted with one or more substituents selected from the group consisting of hydrogen, C ₁ -C ₆ alkyl and acetyl;
R ₃ is independently hydrogen, halogen or C ₁ -C ₆ alkoxy;
R ₄ is substituted or unsubstituted and substituted or unsubstituted is substituted with one or more substituents selected from the group consisting of,
The above substituted and In which at least one hydrogen in the substituent is independently substituted with at least one substituent selected from the group consisting of hydrogen, halogen, halo C ₁ -C ₆ alkyl, CN and C ₁ -C ₆ alkyl,
n is an integer from 0 to 3;
A pharmaceutical composition for preventing or treating colon cancer, characterized by: A pharmaceutical composition for preventing or treating colon cancer, comprising a compound represented by the following chemical formula 3, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof as an active ingredient:
[Chemical Formula 3]

In the above chemical formula 3,
R ₁ is hydrogen, halogen, hydroxy, amino, amide, CN, halo C ₁ -C ₆ alkyl, NO ₂ , C ₁ -C ₆ alkyl, C ₁ -C ₁₀ alkoxy, acetyl, acetamido, C ₁ -C ₆ alkylamino, di(C ₁ -C ₆ ) Substituted with one or more substituents selected from the group consisting of alkylamino, C ₁ -C ₆ alkylcarbonylamino, phenyl, thiophenyl, and morpholinyl;
R ₂ is substituted with one or more substituents selected from the group consisting of hydrogen, halogen, hydroxy, amide, CN, halo C ₁ -C ₆ alkyl, NO ₂ , C _{1 -C 6} alkyl, C ₁ -C ₁₀ alkoxy, acetyl, C ₁ -C ₆ alkylamino, di(C ₁ -C _{6 )} alkylamino, di(C ₁ -C _{6) alkylamino(C 1 -C 6)} alkyl, phenyl, and SEM(=2-(trimethylsilyl)ethoxymethyl);
R ₃ is independently hydrogen, halogen or C ₁ -C ₁₀ alkoxy;
R ₄ is substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted and substituted or unsubstituted is substituted with one or more substituents selected from the group consisting of,
The above substituted , , , , and is substituted with one or more substituents independently selected from the group consisting of hydrogen, halogen, hydroxy, amide, halo C ₁ -C ₆ alkyl, CN, NO ₂ , C ₁ -C ₆ alkyl, acetyl, carbamoyl, substituted or unsubstituted C ₁ -C ₁₀ alkoxy, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted C ₄ -C ₁₀ heterocycloalkyl,
The above substituted C ₁ -C ₁₀ alkoxy, C ₃ -C ₁₀ cycloalkyl or C ₄ -C ₁₀ heterocycloalkyl is substituted with one or more substituents selected from the group consisting of hydrogen, halogen, hydroxy, amide, halo C ₁ -C ₆ alkyl, CN, NO ₂ , C ₁ -C ₆ alkyl, C ₁ -C ₁₀ alkoxy;
n is an integer from 0 to 3; In paragraph 5,
In the above chemical formula 3,
R ₁ is substituted with one or more substituents selected from the group consisting of hydrogen, amino, amide, halo C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl, acetyl, acetamido, phenyl, thiophenyl, and morpholinyl;
R ₂ is substituted with one or more substituents selected from the group consisting of hydrogen, halo C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl, and acetyl;
R ₃ is independently hydrogen, halogen or C ₁ -C ₆ alkoxy;
R ₄ is substituted or unsubstituted and substituted or unsubstituted is substituted with one or more substituents selected from the group consisting of,
The above substituted and At least one hydrogen in the substituent is independently substituted with at least one substituent selected from the group consisting of hydrogen, halogen, halo C ₁ -C ₆ alkyl, CN, C ₁ -C ₆ alkyl and acetyl;
n is an integer from 0 to 3;
A pharmaceutical composition for preventing or treating colon cancer, characterized by: A pharmaceutical composition for preventing or treating colon cancer, comprising a compound represented by the following chemical formula 4, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof as an active ingredient:
[Chemical Formula 4]

In the above chemical formula 4,
R ₁ is hydrogen, halogen, hydroxy, amino, amide, CN, halo C ₁ -C ₆ alkyl, NO ₂ , C ₁ -C ₆ alkyl, C ₁ -C ₁₀ alkoxy, acetyl, acetamido, C ₁ -C ₆ alkylamino, di(C ₁ -C ₆ ) Substituted with one or more substituents selected from the group consisting of alkylamino, C ₁ -C ₆ alkylcarbonylamino, phenyl, thiophenyl, and morpholinyl;
R ₂ is substituted with one or more substituents selected from the group consisting of hydrogen, halogen, hydroxy, amide, CN, halo C ₁ -C ₆ alkyl, NO ₂ , C _{1 -C 6} alkyl, C ₁ -C ₁₀ alkoxy, acetyl, C ₁ -C ₆ alkylamino, di(C ₁ -C ₆ ) alkylamino, di(C ₁ -C _{6 ) alkylamino (C 1 -C 6)} alkyl, phenyl, and SEM(=2-(trimethylsilyl)ethoxymethyl);
R ₃ is independently hydrogen, halogen or C ₁ -C ₁₀ alkoxy;
R ₄ is substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted and substituted or unsubstituted is substituted with one or more substituents selected from the group consisting of,
The above substituted , , , , and is substituted with one or more substituents independently selected from the group consisting of hydrogen, halogen, hydroxy, amide, halo C ₁ -C ₆ alkyl, CN, NO ₂ , C ₁ -C ₆ alkyl, acetyl, carbamoyl, substituted or unsubstituted C ₁ -C ₁₀ alkoxy, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted C ₄ -C ₁₀ heterocycloalkyl,
The above substituted C ₁ -C ₁₀ alkoxy, C ₃ -C ₁₀ cycloalkyl or C ₄ -C ₁₀ heterocycloalkyl is substituted with one or more substituents selected from the group consisting of hydrogen, halogen, hydroxy, amide, halo C ₁ -C ₆ alkyl, CN, NO ₂ , C ₁ -C ₆ alkyl, C ₁ -C ₁₀ alkoxy;
n is an integer from 0 to 3; In paragraph 7,
In the above chemical formula 4,
R ₁ is substituted with one or more substituents selected from the group consisting of hydrogen, amino, amide, halo C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl, acetyl, acetamido, phenyl, thiophenyl, and morpholinyl;
R ₂ is substituted with one or more substituents selected from the group consisting of hydrogen, halo C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl, and acetyl;
R ₃ is independently hydrogen, halogen or C ₁ -C ₆ alkoxy;
R ₄ is substituted or unsubstituted and substituted or unsubstituted is substituted with one or more substituents selected from the group consisting of,
The above substituted and wherein at least one hydrogen in the substituent is independently substituted with at least one substituent selected from the group consisting of hydrogen, halogen, halo C ₁ -C ₆ alkyl, CN, C ₁ -C ₆ alkyl and acetyl;
n is an integer from 0 to 3;
A pharmaceutical composition for preventing or treating colon cancer, characterized by: A pharmaceutical composition for preventing or treating colon cancer, comprising a compound represented by the following chemical formula 5, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof as an active ingredient:
[Chemical Formula 5]

In the above chemical formula 5,
R ₁ is substituted with one or more substituents selected from the _group consisting of hydrogen, halogen, hydroxy, amino, amide, CN, halo C ₁ -C ₆ alkyl, NO ₂ , C _{1 -C 6} alkyl, C ₁ -C ₁₀ alkoxy, acetyl, acetamido, C 1 -C 6 alkylamino, di(C ₁ -C ₆₎ alkylamino, C ₁ -C ₆ alkylcarbonylamino, phenyl, thiophenyl, and morpholinyl;
R ₂ is substituted with one or more substituents selected from the group consisting of hydrogen, halogen, hydroxy, amide, CN, halo C ₁ -C ₆ alkyl, NO ₂ , C _{1 -C 6} alkyl, C ₁ -C ₁₀ alkoxy, acetyl, C ₁ -C ₆ alkylamino, di(C ₁ -C _{6 )} alkylamino, di(C ₁ -C _{6) alkylamino (C 1 -C 6)} alkyl, phenyl, and SEM(=2-(trimethylsilyl)ethoxymethyl);
R ₃ is independently hydrogen, halogen or C ₁ -C ₆ alkoxy;
R ₄ is substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted , substituted or unsubstituted and substituted or unsubstituted is substituted with one or more substituents selected from the group consisting of,
The above substituted , , , , and is substituted with one or more substituents independently selected from the group consisting of hydrogen, halogen, hydroxy, amide, halo C ₁ -C ₆ alkyl, CN, NO ₂ , C ₁ -C ₆ alkyl, acetyl, carbamoyl, substituted or unsubstituted C ₁ -C ₁₀ alkoxy, substituted or unsubstituted C ₃ -C ₁₀ cycloalkyl, substituted or unsubstituted C ₄ -C ₁₀ heterocycloalkyl,
The above substituted C ₁ -C ₁₀ alkoxy, C ₃ -C ₁₀ cycloalkyl or C ₄ -C ₁₀ heterocycloalkyl is substituted with one or more substituents selected from the group consisting of hydrogen, halogen, hydroxy, amide, halo C ₁ -C ₆ alkyl, CN, NO ₂ , C ₁ -C ₆ alkyl, C ₁ -C ₁₀ alkoxy;
n is an integer from 0 to 3; In paragraph 9,
In the above chemical formula 5,
R ₁ is substituted with one or more substituents selected from the group consisting of hydrogen, amino, amide, halo C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl, acetyl, acetamido, phenyl, thiophenyl, and morpholinyl;
R ₂ is substituted with one or more substituents selected from the group consisting of hydrogen, halo C ₁ -C ₆ alkyl, C ₁ -C ₆ alkyl, and acetyl;
R ₃ is independently hydrogen, halogen or C ₁ -C ₆ alkoxy;
R ₄ is substituted or unsubstituted and substituted or unsubstituted is substituted with one or more substituents selected from the group consisting of,
The above substituted and wherein at least one hydrogen in the substituent is independently substituted with at least one substituent selected from the group consisting of hydrogen, halogen, halo C ₁ -C ₆ alkyl, CN, C ₁ -C ₆ alkyl and acetyl;
n is an integer from 0 to 3;
A pharmaceutical composition for preventing or treating colon cancer, characterized by: A pharmaceutical composition for preventing or treating colon cancer, comprising as an active ingredient a compound selected from the group consisting of the following compounds, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof:
4-Methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide (Compound 1);
4-Methoxy-N-phenyl-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 2);
4-Methoxy-N-(3-methoxyphenyl)-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 3);
3-((4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)phenyl)sulfonamido)benzamide (compound 4);
4-Methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(3-(trifluoromethyl)phenyl)benzenesulfonamide (Compound 5);
N-(3-Cyanophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 6);
N-(3-Fluorophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 7);
N-(4-Fluorophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 8);
N-(2-Fluorophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 9);
N-(3-chlorophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 10);
N-(4-chlorophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 11);
N-(2-chlorophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 12);
N-(2-Cyclopropylphenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 13);
N-(4-hydroxyphenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 14);
N-(3-(2-hydroxyethoxy)phenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 15);
N-(4-(2-hydroxyethoxy)phenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 16);
N-(2-(2-hydroxyethoxy)phenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 17);
4-Methoxy-N-(4-(4-methylpiperazin-1-yl)phenyl)-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 18);
4-Methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(pyridin-3-yl)benzenesulfonamide (Compound 19);
N-(3-fluorophenyl)-3-(4-oxo-1,4-dihydroquinolin-6-yl)benzenesulfonamide (Compound 20);
5-(5-((1H-Pyrazol-1-yl)sulfonyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridine (Compound 21);
5-(2-methoxy-5-((4-methylpiperazin-1-yl)sulfonyl)phenyl)-1H-pyrazolo[3,4-b]pyridine (Compound 22);
4-((4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)phenyl)sulfonyl)morpholine (Compound 23);
N-(3,5-difluorophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 24);
N-(3-chloro-5-fluorophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 25);
4-Methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzamide (Compound 26);
N-(3-Fluorophenyl)-4-methoxy-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzamide (Compound 27);
N-Phenyl-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 28);
3-(1H-Pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide (Compound 29);
N-(3-Fluorophenyl)-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 30);
N-(3-chlorophenyl)-3-(1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 31);
3-(1H-Pyrazolo[3,4-b]pyridin-5-yl)-N-(o-tolyl)benzenesulfonamide (Compound 32);
4-Methoxy-3-(3-methyl-1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide (Compound 33);
4-Methoxy-3-(1-phenyl-1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide (Compound 34);
N-(5-(2-methoxy-5-(N-(m-tolyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 35);
N-(5-(4-fluoro-3-(N-(m-tolyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 36);
N-(5-(2-Fluoro-5-(N-(m-tolyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 37);
N-(5-(3-fluoro-5-(N-(m-tolyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 38);
N-(5-(2,4-difluoro-5-(N-(m-tolyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 39);
N-(5-(2-Fluoro-3-(N-(m-tolyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 40);
N-(5-(2-methoxy-5-(N-(p-tolyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 41);
N-(5-(2-methoxy-5-(N-(o-tolyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 42);
N-(5-(2-methoxy-5-(N-phenylsulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 43);
N-(5-(5-(N-(3-fluorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 44);
N-(5-(4-fluoro-3-(N-(3-fluorophenyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 45);
N-(5-(2-Fluoro-5-(N-(3-fluorophenyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 46);
N-(5-(3-fluoro-5-(3-fluorophenyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 47);
N-(5-(2,4-difluoro-5-(N-(3-fluorophenyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 48);
N-(5-(2-Fluoro-3-(N-(3-fluorophenyl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 49);
N-(5-(5-(N-(4-fluorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 50);
N-(5-(5-(N-(2-fluorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 51);
N-(5-(5-(N-(3-chlorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 52);
N-(5-(5-(N-(4-chlorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 53);
N-(5-(5-(N-(2-chlorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 54);
N-(5-(5-(N-(3-hydroxyphenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 55);
N-(5-(5-(N-(4-hydroxyphenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 56);
N-(5-(2-methoxy-5-(N-(pyridin-3-yl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 57);
N-(5-(2-methoxy-5-(N-(pyridin-2-yl)sulfamoyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 58);
N-(5-(2-methoxy-5-(morpholinosulfonyl)phenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 59);
N-(5-(5-(N-(3,5-difluorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 60);
N-(5-(5-(N-(3-chloro-5-fluorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-yl)acetamide (Compound 61);
N-(3-Fluorophenyl)-4-methoxy-3-(3-phenyl-1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 62);
N-(3-Fluorophenyl)-4-methoxy-3-(3-(thiophen-3-yl)-1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 63);
4-Methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide (Compound 64);
4-Methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(p-tolyl)benzenesulfonamide (Compound 65);
4-Methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(o-tolyl)benzenesulfonamide (Compound 66);
4-Methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)-N-phenylbenzenesulfonamide (Compound 67);
N-(3-Fluorophenyl)-4-methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 68);
N-(2-Fluorophenyl)-4-methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 69);
N-(2-Chlorophenyl)-4-methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 70);
N-(3-Hydroxyphenyl)-4-methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)benzenesulfonamide (Compound 71);
4-((4-methoxy-3-(3-morpholino-1H-pyrazolo[3,4-b]pyridin-5-yl)phenyl)sulfonyl)morpholine (Compound 72);
4-Methoxy-3-(1-phenyl-1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide (Compound 73);
3-(1-(2-(Dimethylamino)ethyl)-1H-pyrazolo[3,4-b]pyridin-5-yl)-4-methoxy-N-(m-tolyl)benzenesulfonamide (Compound 74);
4-Methoxy-3-(1-methyl-1H-pyrazolo[3,4-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide (Compound 75);
3-(1-Acetyl-1H-pyrazolo[3,4-b]pyridin-5-yl)-4-methoxy-N-(m-tolyl)benzenesulfonamide (Compound 76);
N-((3-(1-Acetyl-1H-pyrazolo[3,4-b]pyridin-5-yl)-4-methoxyphenyl)sulfonyl)-N-(m-tolyl)acetamide (Compound 77);
4-((3-(1H-indazol-5-yl)-4-methoxyphenyl)sulfonyl)morpholine (Compound 78);
3-(1H-indazol-5-yl)-4-methoxy-N-(m-tolyl)benzenesulfonamide (Compound 79);
N-(3-fluorophenyl)-3-(1H-indazol-5-yl)-4-methoxybenzenesulfonamide (Compound 80);
2,4-Difluoro-N-(3-fluorophenyl)-5-(1H-indazol-5-yl)benzenesulfonamide (Compound 81);
N-(5-(5-(N-(3-fluorophenyl)sulfamoyl)-2-methoxyphenyl)-1H-indazol-3-yl)acetamide (Compound 82);
N-(5-(2,4-difluoro-5-(N-(3-fluorophenyl)sulfamoyl)phenyl)-1H-indazol-3-yl)acetamide (Compound 83);
3-(1-Acetyl-3-amino-1H-indazol-5-yl)-N-(3-fluorophenyl)-4-methoxybenzenesulfonamide (Compound 84);
5-(1-Acetyl-3-amino-1H-indazol-5-yl)-2,4-difluoro-N-(3-fluorophenyl)benzenesulfonamide (Compound 85);
4-((4-methoxy-3-(1H-pyrrolo[2,3-b]pyridin-5-yl)phenyl)sulfonyl)morpholine (Compound 86);
4-Methoxy-3-(1H-pyrrolo[2,3-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide (Compound 87);
N-(3-Fluorophenyl)-4-methoxy-3-(1H-pyrrolo[2,3-b]pyridin-5-yl)benzenesulfonamide (Compound 88);
4-Methoxy-3-(3-morpholino-1H-pyrrolo[2,3-b]pyridin-5-yl)-N-(m-tolyl)benzenesulfonamide (Compound 90);
4-Methoxy-3-(3-morpholino-1H-pyrrolo[2,3-b]pyridin-5-yl)-N-(o-tolyl)benzenesulfonamide (Compound 91);
4-Methoxy-3-(3-morpholino-1H-pyrrolo[2,3-b]pyridin-5-yl)-N-phenylbenzenesulfonamide (Compound 92);
N-(2-Fluorophenyl)-4-methoxy-3-(3-morpholino-1H-pyrrolo[2,3-b]pyridin-5-yl)benzenesulfonamide (Compound 93);
N-(2-Chlorophenyl)-4-methoxy-3-(3-morpholino-1H-pyrrolo[2,3-b]pyridin-5-yl)benzenesulfonamide (Compound 94);
N-(5-(2-methoxy-5-(N-phenylsulfamoyl)phenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)acetamide (Compound 95);
N-(5-(2,4-difluoro-5-(N-(3-fluorophenyl)sulfamoyl)phenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl)acetamide (Compound 96);
3-(1H-Indol-5-yl)-4-methoxy-N-(m-tolyl)benzenesulfonamide (Compound 97);
N-(3-Fluorophenyl)-3-(1H-indol-5-yl)-4-methoxybenzenesulfonamide (Compound 98);
4-Methoxy-N-(m-tolyl)-3-(3-((2-(trimethylsilyl)ethoxy)methyl)-3H-imidazo[4,5-b]pyridin-6-yl)benzenesulfonamide (Compound 99);
3-(3H-Imidazo[4,5-b]pyridin-6-yl)-4-methoxy-N-(m-tolyl)benzenesulfonamide (Compound 100);
N-(3-Fluorophenyl)-4-methoxy-3-(3-((2-(trimethylsilyl)ethoxy)methyl)-3H-imidazo[4,5-b]pyridin-6-yl)benzenesulfonamide (Compound 101);
N-(3-Fluorophenyl)-3-(3H-imidazo[4,5-b]pyridin-6-yl)-4-methoxybenzenesulfonamide (Compound 102);
4-Methoxy-3-(4-oxo-1,4-dihydroquinolin-6-yl)-N-(m-tolyl)benzenesulfonamide (Compound 103);
N-(3-fluorophenyl)-4-methoxy-3-(4-oxo-1,4-dihydroquinolin-6-yl)benzenesulfonamide (Compound 104); and
3-(4-Oxo-1,4-dihydroquinolin-6-yl)-N-(m-tolyl)benzenesulfonamide (Compound 105). In any one of claims 1 to 11,
A pharmaceutical composition for preventing or treating colon cancer, characterized in that the colon cancer includes rectal cancer, colon cancer, and anal cancer. In Article 12,
The pharmaceutical composition for preventing or treating colon cancer is characterized in that it blocks IRP2 from binding to IRE. In any one of claims 1 to 11,
A pharmaceutical composition for preventing or treating colon cancer, characterized in that it additionally comprises a pharmaceutically acceptable carrier, diluent or excipient.