WO2025037870A1 - Benzofuran compound and pharmaceutical composition comprising same for preventing or treating cancer diseases and cancer metastasis - Google Patents

Abstract

The present invention relates to a novel benzofuran compound and a pharmaceutical composition comprising same for preventing or treating cancer. The compound according to the present invention or a pharmaceutically acceptable salt thereof binds to hnRNPA2B1 to inhibit the translation of HIF-1α, thus exhibiting an excellent prophylactic and therapeutic effect on cancer diseases of various carcinomas. In particular, the present invention exhibits an effect of having excellent anticancer activity through the effect of inhibiting cancer metastasis as well as the effect of inhibiting cell death.

Description

Benzofuran compound and pharmaceutical composition containing the same for preventing or treating cancer disease and cancer metastasis

The present invention relates to a novel benzofuran compound and a pharmaceutical composition containing the same for preventing or treating cancer and cancer metastasis.

Hypoxia-inducible factor-1 (HIF-1) is a transcription factor involved in cellular adaptation and regulates various biological processes such as cell proliferation, angiogenesis, cellular energy metabolism, apoptosis resistance, and metastasis. HIF-1 is a heterodimeric protein composed of HIF-1α and HIF-1β subunits. HIF-1β is a protein that is normally present in cells called aryl hydrocarbon nuclear translocator (ARNT), and HIF-1α is a protein that is expressed and activated by intracellular O ₂ concentration. More specifically, HIF-1α is degraded under normal oxygen concentration, but stably accumulates in cells under hypoxic conditions and activates the expression of various genes that help cells adapt to hypoxia. Therefore, HIF-1α plays an important role in cell adaptation under hypoxic conditions.

Hypoxia in cancer, especially in solid cancer, is a common phenomenon, and solid cancer cells adapt to these hypoxic conditions through various genetic changes, which makes cancer cells more malignant and resistant to anticancer drugs. HIF-1 is known as the most important molecule that regulates the adaptation of cancer cells to these hypoxic conditions, and in particular, the amount of HIF-1α protein and the prognosis of cancer patients are known to be closely correlated. In addition, it is known that active activity of HIF-1 protein promotes tumor growth and interferes with the effectiveness of chemotherapy or radiotherapy, worsening the condition of patients.

Meanwhile, Ras-associated factor-1 (RAF-1) belongs to the RAF protein kinase family, also known as C-Raf. It is an important part of the human RAS/RAF/MEK/ERK signaling pathway and is closely related to the regulation of cell proliferation and differentiation.

High expression of RAF-1 is positively correlated with the occurrence, development, and clinical prognosis of various cancers, and RAF-1 is recognized as a cancer metastasis drug target.

Accordingly, the inventors of the present invention prepared a novel benzofuran compound and confirmed that the compound can prevent or treat cancer and cancer metastasis by effectively inhibiting the accumulation of HIF-1α and the expression of RAF-1, thereby completing the present invention.

The purpose of the present invention is to provide a novel benzofuran compound or a pharmaceutically acceptable salt thereof.

The purpose of the present invention is to provide a novel benzofuran compound and a pharmaceutical composition containing the same for preventing or treating cancer.

In order to avoid confusion due to overlapping content, overlapping content will be omitted in the following description. In other words, the content of the invention is not limited to the content below, and the content of the invention should be interpreted according to the content of the entire invention.

The term "halogen" in the present invention means a substituent selected from fluorine (F), chloro (Cl), bromo (Br) and iodo (I). Preferably, the halogen may be fluorine.

In the present invention, “C _x -C _y ” means having x or more and y or less carbon atoms.

In the present invention, the term "substituted" refers to a moiety having a substituent replacing a hydrogen on one or more carbon atoms of the main chain. "Substituted" or "substituted with ~" is defined to include the implicit condition that such substitution is dependent on the permissibility of the substituted atom and the substituent, and that the substitution leads to a stable compound, for example, a compound that does not undergo spontaneous transformation by rearrangement, cyclization, elimination, etc.

In the present invention, "single bond" means that two connected radicals are directly connected. For example, if L represents a single bond in A-L-Z, this structure is essentially A-Z.

The term "C ₁ -C ₆ alkyl" as used herein means a straight or branched C ₁ -C ₆ saturated hydrocarbon group, such as, for example, methyl, ethyl, propyl, isopropyl, butyl, pentyl, hexyl, and the like. Preferred alkyl groups contain about 1, 2, 3, 4, 5 or 6 carbon atoms in the chain. A side chain means that one or more lower alkyl groups, for example, methyl, ethyl or propyl, are attached to a linear alkyl chain. "Lower alkyl" means a group having from about 1 to about 6 carbon atoms in the chain, which may be straight or branched. "Alkyl" may be unsubstituted or optionally substituted by one or more substituents, which may be the same or different. Each substituent can be halogen, alkyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, cyano, hydroxy, alkoxy, alkylthio, amino, carboxy, etc. Each of these substituents can follow any of the definitions specified for each substituent mentioned herein.

The term "C ₆ -C ₁₂ aryl" as used herein refers to an aromatic hydrocarbon containing 6 to 12 carbon atoms. For example, it can refer to a ring system such as monocyclic (e.g., phenyl); bicyclic (e.g., indenyl, naphthalenyl, tetrahydronaphthyl, tetrahydroindenyl). Preferably, aryl can be a phenyl group having a chemical formula of C ₆ H ₅ and having 6 carbon atoms arranged in a cyclic ring structure. The phenyl group is very stable and is a type of aromatic hydrocarbon found in many organic compounds. In addition, the above aryl may be substituted or unsubstituted, and when substituted, the hydrogen at the ortho, meta, or para position of the phenyl ring may be substituted with a halogen atom, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, -OC ₁ -C ₆ alkyl, -OC ₁ -C ₆ haloalkyl, oxo(=O), -OH, -CN, -N-(C ₁ -C ₆ alkyl) ₂ or -NH ₂ .

The term "C ₃ -C ₁₂ heteroaryl" as used herein refers to an optionally substituted aromatic ring containing 3 to 12 carbon atoms, wherein at least one of the ring carbon atoms is replaced by a heteroatom selected from oxygen (O), nitrogen (N) and sulfur (S), or an aromatic ring (e.g., a bicyclic or tricyclic ring system) fused to one or more rings such as a heteroaryl ring, an aryl ring, a heterocyclic ring, or a carbocyclic ring, each of which may have an optional substituent.

In the present invention, "C ₃ -C ₈ heterocycloalkyl" includes a saturated monocyclic or polycyclic heterocycle containing 1 to 4 heteroatoms independently selected from nitrogen (N), oxygen (O) and sulfur (S), or a ring structure in which two or more rings share one or more pairs of carbon atoms (e.g., a fused ring, a spiro ring, a bridged ring, etc.). In addition, the heterocycloalkyl may be substituted or unsubstituted, and if substituted, may be substituted with a halogen atom, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, -OC ₁ -C ₆ alkyl, -OC ₁ -C ₆ haloalkyl, oxo (=O), -OH, -CN, -N-(C ₁ -C ₆ alkyl) ₂ or -NH ₂ . When nitrogen is present in the heterocycloalkyl ring, it may exist in an oxidized state (i.e., N ⁺ -O-) as long as the properties of the adjacent atoms and groups permit. Examples include piperidinyl N-oxide and morpholinyl-N-oxide. Additionally, when sulfur is present in the heterocycloalkyl ring, it may exist in an oxidized state (i.e., S ⁺ -O- or -SO ₂ -) as long as the properties of the adjacent atoms and groups permit. Examples include thiomorpholine S-oxide and thiomorpholine S,S-dioxide. Additionally, one ring of the polycyclic heterocycloalkyl group can be aromatic (e.g., aryl or heteroaryl) if the polycyclic heterocycloalkyl group is attached to the parent structure through a non-aromatic carbon or nitrogen atom.

In the present invention, “alkoxy” is an alkyl group (a chain of carbon and hydrogen) bonded to oxygen, and means a straight or branched chain alkoxy of C ₁ -C ₆ such as methoxy, ethoxy, etc.

In the present invention, “amino” means a form in which hydrogen is bonded to a nitrogen atom, and this hydrogen atom may be substituted with a C ₁ -C ₆ straight chain or branched chain alkyl, aryl, etc.

In the present invention, “carboxy” means a substituent represented by -COOH or -CO ₂ H.

In the present invention, “haloalkyl” means an alkyl group substituted with one or more halogens (e.g., fluoride, chloride, bromide, or iodide).

In the present invention, “halobenzyl” means a benzyl group substituted with one or more halogens (e.g., fluoride, chloride, bromide, or iodide).

In addition, terms and abbreviations used in this specification have their original meanings unless otherwise defined.

A compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof

In one aspect of the present invention, a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof is provided.

[Chemical Formula 1]

In the above chemical formula 1,

R ₁ is a hydrogen atom, -OH, or C ₁ -C ₆ alkyl,

는

이고,

Is

And,

,

, -NHR₇ 또는 산소, 질소 및 황으로부터 선택되는 헤테로원자를 포함하는 것이고,R ₆ is independently a hydrogen atom, a halogen atom, -OH, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy, -R ₂ OH,

,

, -NHR ₇ or a heteroatom selected from oxygen, nitrogen and sulfur,

R ₂ is C ₁ -C ₆ alkyl,

R ₃ is a hydrogen atom or C ₁ -C ₆ alkyl,

R ₄ and R ₅ are the same or different, and are each independently a hydrogen atom, a substituted or unsubstituted C ₁ -C ₆ alkyl, a substituted or unsubstituted C ₃ -C ₈ heterocycloalkyl, or R ₄ and R ₅ are connected to each other together with one or more carbon atoms or nitrogen atoms to form a substituted or unsubstituted C ₃ -C ₈ heterocycloalkyl, wherein the substitution is substitution with at least one or more C ₁ -C ₃ alkyl, C ₆ -C ₁₂ aryl, -C(=O)O-(C ₁ -C ₆ alkyl), or halobenzyl,

R ₇ is a hydrogen atom or C ₁ -C ₆ alkyl,

n is an integer from 0 to 5, and when n is 2 or greater, R ₆ are equal or different.

According to one embodiment of the present invention, in the chemical formula 1,

는

이고,

Is

And,

,

, -NHR₇ 또는 산소, 질소 및 황으로부터 선택되는 헤테로원자를 포함하는 것이고,R6a, R6b, R6c and R6e are the same or different and each independently represents a hydrogen atom, a halogen atom, -OH, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy, -R ₂ OH,

,

, -NHR ₇ or a heteroatom selected from oxygen, nitrogen and sulfur,

,

, -NHR₇ 또는 산소, 질소 및 황으로부터 선택되는 헤테로원자를 포함하는 것일 수 있다.R6d is hydrogen atom, halogen atom, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy, -R ₂ OH,

,

, -NHR ₇ or may contain a heteroatom selected from oxygen, nitrogen and sulfur.

According to one embodiment of the present invention, when R ₁ in the chemical formula 1 is methyl, R6b and R6d are not OH.

In addition, according to one embodiment of the present invention, the compound represented by the chemical formula 1 may be a compound represented by the following chemical formula 2.

[Chemical formula 2]

In the above chemical formula 2,

R ₁ is a hydrogen atom, -OH, or C ₁ -C ₆ alkyl,

R ₄ and R ₅ are the same or different, and are each independently a hydrogen atom, a substituted or unsubstituted C ₁ -C ₆ alkyl, a substituted or unsubstituted C ₃ -C ₈ heterocycloalkyl, or R ₄ and R ₅ are connected to each other together with one or more carbon atoms or nitrogen atoms to form a substituted or unsubstituted C ₃ -C ₈ heterocycloalkyl, wherein the substitution is substitution with at least one or more C ₁ -C ₃ alkyl, C ₆ -C ₁₂ aryl, -C(=O)O-(C ₁ -C ₆ alkyl), or halobenzyl,

,

, -NHR₇ 또는 산소, 질소 및 황으로부터 선택되는 헤테로원자를 포함하는 것이고,R ₆ is hydrogen atom, halogen atom, -OH, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy, -R ₂ OH,

,

, -NHR ₇ or a heteroatom selected from oxygen, nitrogen and sulfur,

R ₂ is C ₁ -C ₆ alkyl,

R ₇ is a hydrogen atom or C ₁ -C ₆ alkyl,

n is an integer from 0 to 4, and when n is 2 or greater, R ₆ are identical or different.

According to one embodiment of the present invention, in the chemical formula 1, R ₁ is hydrogen, OH, methyl, ethyl, or isobutyl,

는

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

, 또는

일 수 있다.

Is

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

, or

It could be.

는

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

, 또는

일 수 있다.More specifically,

Is

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

, or

It could be.

는

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

, 또는

일 수 있다.According to another embodiment of the present invention, R ₁ is methyl,

Is

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

, or

It could be.

는

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

, 또는

일 수 있다.More specifically,

Is

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

, or

It could be.

According to a specific example of the present invention, the compound of the chemical formula 1 may be any one selected from the group consisting of compounds of the following Table 1:

[Table 1]

That is, according to a specific example of the present invention, the compound of the chemical formula 1 may be any one selected from the group consisting of the following compounds:

5-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)benzene-1,3-diol;

4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)phenol;

4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)benzene-1,2-diol;

3-methoxy-5-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)phenol;

Methyl 4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)benzoate;

4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)benzoic acid;

[4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)phenyl](4-methylpiperazin-1-yl)methanone;

2-Fluoro-4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)phenol;

N,N-Dimethyl-4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)benzamide;

2-(4-fluorophenyl)-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran;

2-(3,5-difluorophenyl)-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran;

[4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)phenyl](piperidin-1-yl)methanone;

4-(7-Methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)-N-(1-methylpiperidin-4-yl)benzamide;

N-Benzyl-4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)benzamide;

tert-Butyl 4-[4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)benzoyl]piperazine-1-carboxylate;

[4-(4-fluorobenzyl)piperazin-1-yl][4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)phenyl]methanone;

[4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)phenyl](piperazin-1-yl)methanone;

3-[7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)-N-(1-methylpiperidin-4-yl)benzamide;

2-Methyl-4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)-N-(1-methylpiperidin-4-yl)benzamide; and

3-Methyl-4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)-N-(1-methylpiperidin-4-yl)benzamide.

The compound represented by chemical formula 1 of the present invention or a pharmaceutically acceptable salt thereof may target hnRNPA2B1.

In the present invention, "hnRNPA2B1" refers to a heterogeneous nuclear ribonucleoprotein A2B1, which is localized in the nucleus and cytoplasm and is a member of the RNA binding protein group associated with early pre-mRNA. The expression of hnRNPA2B1 is increased in cancer patients and is closely related to apoptosis. hnRNPA2B1 is a putative proto-oncogene in glioblastoma and regulates several tumor suppressors and oncogenes. Under hypoxic conditions, the expression of HIF-1α protein is efficiently promoted through the interaction of hnRNPA2B1.

Figure 1 is a schematic diagram showing the role of MO-2097 (compound 1a of Example 1) in inhibiting HIF-1α mRNA translation in cancer tissues mediated by hnRNPA2B1.

According to FIG. 1, the compound represented by the chemical formula 1 of the present invention or a pharmaceutically acceptable salt thereof can exhibit excellent therapeutic efficacy against cancer by binding to hnRNPA2B1 and inhibiting the translation of HIF-1α.

The compound represented by the chemical formula 1 of the present invention or a pharmaceutically acceptable salt thereof can bind to hnRNPA2B1, inhibit the activity of RAF-1, and prevent and treat cancer metastasis.

In the present invention, pharmaceutically acceptable salts mean salts commonly used in the pharmaceutical industry, and include, for example, inorganic ion salts manufactured with calcium, potassium, sodium, and magnesium; inorganic acid salts manufactured with hydrochloric acid, nitric acid, phosphoric acid, hydrobromic acid, iodic acid, perchloric acid, and sulfuric acid; organic acid salts manufactured with acetic acid, trifluoroacetic acid, citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, and hydroiodic acid; sulfonic acid salts manufactured with methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, and naphthalenesulfonic acid; Amino acid salts manufactured from glycine, arginine, lysine, etc.; and amine salts manufactured from trimethylamine, triethylamine, ammonia, pyridine, picoline, etc.; However, the types of salts meant in the present invention are not limited by these listed salts.

In addition, the compound represented by the chemical formula 1 of the present invention includes not only pharmaceutically acceptable salts, but also all salts, hydrates, and solvates that can be prepared by conventional methods.

In another embodiment of the present invention, a pharmaceutical composition comprising a compound represented by formula 1 as defined in any embodiment described herein or a pharmaceutically acceptable salt thereof is provided.

[Chemical Formula 1]

In the above chemical formula 1,

R ₁ is a hydrogen atom, -OH, or C ₁ -C ₆ alkyl,

,

, -NHR₇ 또는 산소, 질소 및 황으로부터 선택되는 헤테로원자를 포함하는 것이고,R ₆ is independently a hydrogen atom, a halogen atom, -OH, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy, -R ₂ OH,

,

, -NHR ₇ or a heteroatom selected from oxygen, nitrogen and sulfur,

R ₂ is C ₁ -C ₆ alkyl,

R ₃ is a hydrogen atom or C ₁ -C ₆ alkyl,

R ₄ and R ₅ are the same or different, and are each independently a hydrogen atom, a substituted or unsubstituted C ₁ -C ₆ alkyl, a substituted or unsubstituted C ₃ -C ₈ heterocycloalkyl, or R ₄ and R ₅ are connected to each other together with one or more carbon atoms or nitrogen atoms to form a substituted or unsubstituted C ₃ -C ₈ heterocycloalkyl, wherein the substitution is substitution with at least one or more C ₁ -C ₃ alkyl, C ₆ -C ₁₂ aryl, -C(=O)O-(C ₁ -C ₆ alkyl), or halobenzyl,

R ₇ is a hydrogen atom or C ₁ -C ₆ alkyl,

n is an integer from 0 to 5, and when n is 2 or greater, R ₆ are equal or different.

In another embodiment of the present invention, the present invention provides a pharmaceutical composition for preventing or treating cancer, comprising a compound represented by the following chemical formula 3 or a pharmaceutically acceptable salt thereof.

[Chemical Formula 3]

In another embodiment of the present invention, a pharmaceutical composition for preventing or treating cancer is provided, comprising a compound represented by formula 1 as defined in any embodiment described in the present invention or a pharmaceutically acceptable salt thereof.

In the present invention, “prevention” means any act of inhibiting or delaying the onset of cancer formation or cancer metastasis by administering a composition, and “treatment” means any act of improving or beneficially changing the symptoms of a disease by administering a composition.

In the present invention, the cancer may be any one selected from the group consisting of colon cancer, breast cancer, brain cancer, neurological cancer, lung cancer, small cell lung cancer, stomach cancer, liver cancer, blood cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, cutaneous or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, anal cancer, colon cancer, breast cancer, fallopian tube carcinoma, endometrial carcinoma, cervical cancer, vaginal cancer, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, renal cell carcinoma, renal pelvic carcinoma, CNS tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma, and pituitary adenoma.

Preferably, the cancer may be any one selected from the group consisting of colon cancer, uterine cancer, endometrial cancer and cervical cancer.

The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier, and may be formulated in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods.

The pharmaceutically acceptable carriers include, but are not limited to, those commonly used in the art, such as lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil.

In addition, the pharmaceutical composition of the present invention may include, but is not limited to, diluents or excipients such as fillers, bulking agents, binders, wetting agents, disintegrants, surfactants, and other pharmaceutically acceptable additives.

When the pharmaceutical composition of the present invention is formulated as an oral solid preparation, it includes tablets, pills, powders, granules, capsules, etc., and such solid preparations may include at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc., and include, but are not limited to, lubricants, such as magnesium stearate and talc.

When the pharmaceutical composition of the present invention is formulated as an oral liquid, it includes a suspension, an oral solution, an emulsion, a syrup, etc., and includes, but is not limited to, a diluent such as water or liquid paraffin, a wetting agent, a sweetener, an air freshener, a preservative, etc.

When the pharmaceutical composition of the present invention is formulated for parenteral use, it includes a sterile aqueous solution, a non-aqueous solvent, a suspension, an emulsion, a lyophilized preparation, and a suppository. Non-aqueous solvents and suspensions include, but are not limited to, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases include, but are not limited to, witepsol, macrogol, Tween 61, cacao butter, laurin butter, and glycerogelatin.

Other carrier materials and administration methods known in the pharmaceutical art may also be used. The pharmaceutical compositions of the present invention may be prepared by any well-known pharmaceutical technique, such as effective formulation and administration procedures.

The composition may be administered in single or multiple doses in a pharmaceutically effective amount. The term "pharmaceutically effective amount" of the present invention means an amount sufficient to prevent or treat a disease at a reasonable benefit/risk ratio applicable to medical prevention or treatment, and the effective dosage level may be determined according to factors including the severity of the disease, the activity of the drug, the patient's age, weight, health, sex, the patient's sensitivity to the drug, the time of administration of the composition of the present invention used, the route of administration and the excretion rate, the treatment period, drugs combined with or co-used with the composition of the present invention used, and other factors well known in the medical field. For example, the compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof may be administered at 0.0001 to 100 mg/kg per day, and the administration may be administered once a day or in several divided doses.

The pharmaceutical composition of the present invention can be administered to mammals such as rats, mice, livestock, and humans by various routes, for example, but not limited to, oral administration, intrathecal, intra-auricular, intraperitoneal or intravenous, intramuscular, subcutaneous, intrauterine epidural, sublingual, or intracerebrovascular injection.

The pharmaceutical composition of the present invention may contain 0.01 to 95 wt%, preferably 1 to 80 wt%, of the compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof based on the total weight of the composition.

The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. In addition, the pharmaceutical composition of the present invention may be administered singly or in multiple doses. It is important to administer an amount that can achieve the maximum effect with the minimum amount without side effects by taking all of the above factors into consideration, and this can be easily determined by those skilled in the art.

The term "subject" of the present invention includes an animal or human whose symptoms can be improved by administration of the pharmaceutical composition according to the present invention. By administering the therapeutic composition according to the present invention to a subject, cancer can be effectively prevented and treated.

The term "administration" of the present invention means introducing a given substance into a human or animal by any appropriate method, and the route of administration of the therapeutic composition according to the present invention may be oral or parenteral administration through any common route as long as it can reach the target tissue. In addition, the therapeutic composition according to the present invention may be administered by any device through which the active ingredient can move to the target cell.

The present invention also includes the following embodiments:

A compound of formula 1 as defined in any embodiment described herein for use as a medicament, or a pharmaceutically acceptable salt thereof;

A compound of formula 1 as defined in any embodiment described herein, or a pharmaceutically acceptable salt thereof, for use in the prevention or treatment of cancer discussed in the present invention;

A compound of formula 1 as defined in any embodiment described herein, or a pharmaceutically acceptable salt thereof, for use in the prevention or treatment of cancer metastasis discussed in the present invention;

A method for treating cancer, comprising administering to a subject in need thereof a therapeutically effective amount of a compound represented by formula 1 or a pharmaceutically acceptable salt thereof, as defined in any embodiment described herein;

A method for inhibiting cancer metastasis, comprising administering to a subject in need thereof a therapeutically effective amount of a compound represented by formula 1 or a pharmaceutically acceptable salt thereof as defined in any embodiment described herein;

Use of a compound represented by formula 1 or a pharmaceutically acceptable salt thereof as defined in any embodiment described in the present invention for the manufacture of a medicament for the prevention or treatment of cancer; or

Use of a compound represented by formula 1 or a pharmaceutically acceptable salt thereof as defined in any embodiment described in the present invention for the manufacture of a medicament for preventing or treating cancer metastasis.

Any of the examples or pharmaceutically acceptable salts thereof may be claimed individually or together as a group in any combination with any number of each and every embodiment described herein.

The term "subject" of the present invention means any animal that has developed or may develop cancer, and typically may be an animal that can exhibit a beneficial effect by treatment with the compound represented by Chemical Formula 1 of the present invention, or a pharmaceutically acceptable salt thereof, but includes, without limitation, any subject that has symptoms of cancer or is likely to have such symptoms. As described above, by administering the pharmaceutical composition of the present invention to a subject, the cancer can be effectively prevented or treated. The pharmaceutical composition of the present invention can be administered as an individual therapeutic agent, or can be administered in combination with existing cancer therapeutic agents, and can be administered sequentially or simultaneously with the existing therapeutic agents.

In another embodiment of the present invention, a food composition for preventing or improving cancer is provided, comprising a compound represented by chemical formula 1 as defined in any embodiment described in the present invention or a pharmaceutically acceptable salt thereof.

In the present invention, “improvement” means any act of improving cancer by administering a composition according to the present invention.

The food composition of the present invention may be, for example, any one selected from among noodles, gum, dairy products, ice cream, meat, grains, caffeinated beverages, general beverages, chocolate, bread, snacks, confectionery, candy, pizza, jelly, alcoholic beverages, alcohol, vitamin complexes, and other health supplements, but is not limited thereto.

When the food composition of the present invention is used as a food additive, it can be added as is or used together with other foods or food ingredients, and can be used appropriately according to a conventional method.

The food composition of the present invention includes a health functional food.

The above “health functional food” refers to food manufactured and processed using raw materials or ingredients with functionality useful to the human body according to Act No. 6727 on Health Functional Foods, and “functionality” refers to consumption for the purpose of obtaining a useful effect for health purposes, such as regulating nutrients for the structure and function of the human body or physiological effects.

The food composition of the present invention may include additional ingredients. For example, it may include biotin, folate, pantothenic acid, vitamins A, C, D, E, B1, B2, B6, B12, niacin, etc. In addition, it may include minerals such as chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), zinc (Zn), iron (Fe), calcium (Ca), etc. In addition, it may include amino acids such as cysteine, valine, lysine, and tryptophan. In addition, food additives such as preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), coloring agents (tar color, etc.), coloring agents (sodium nitrite, sodium nitrate, etc.), bleaching agents (sodium sulfite), bactericides (bleaching powder and high-purity bleaching powder, sodium hypochlorite, etc.), leavening agents (alum, D-potassium hydrogentartrate, etc.), reinforcing agents, emulsifiers, thickeners (glucose), film-forming agents, antioxidants (butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), etc.), seasonings (MSG, monosodium glutamate, etc.), sweeteners (dulcin, cyclamate, saccharin, sodium, etc.), flavorings (vanillin, lactones, etc.), gum bases, antifoaming agents, solvents, and improvers can be added. The above additives can be selected depending on the type of food and used in an appropriate amount.

The compound represented by chemical formula 1 of the present invention or a pharmaceutically acceptable salt thereof binds to hnRNPA2B1 and inhibits the translation of HIF-1α, thereby exhibiting excellent effects in the prevention or treatment of cancer for various types of cancer. In particular, it has excellent anticancer effects by inhibiting cancer metastasis together with inhibition of cell death.

Therefore, the pharmaceutical composition for preventing and treating cancer comprising the compound represented by the chemical formula 1 of the present invention can be usefully used as a cancer treatment agent in terms of lowering cancer cell survival rate and inhibiting cancer metastasis.

Figure 1 is a schematic diagram showing the role of MO-2097 (compound 1a of Example 1) in inhibiting HIF-1α mRNA translation in cancer tissues mediated by hnRNPA2B1.

Figure 2 confirms the effect of MO-2097 on HIF-1α protein expression.

Figure 3 shows the results of evaluating cell death by FASC analysis of MO-2097.

Figure 4 shows the results of the Live cell/Apoptosis cells staining experiment of MO-2097.

Figure 5 shows the results of measuring the binding affinity of hnRNPA2B1 for MO-2097 and Moracin G, respectively.

Figure 6 shows the results of a biotin pull-down assay using HeLa CCL2 lysate to investigate the interaction between MO-2097 and hnRNPA2B1 protein under biochemical conditions.

Figure 7 shows the decrease in nuclear HIF-1α protein in HeLa CCL2 cells treated with the hnRNPA2B1-binding compound MO-2097 in HeLa CCL2 cells exposed to hypoxic conditions.

Figure 8 shows the decrease in nuclear HIF-1α protein in HeLa CCL2 cells treated with the hnRNPA2B1-binding compound MO-2097 in HCT116 cells exposed to hypoxic conditions.

Figure 9 shows the results of transcription/translation analysis to confirm the inhibitory effect of MO-2097 on HIF-1α protein expression.

Figure 10 shows the results of observing the expression of HIF-1α target genes, including HK, MRP1, SLC1A5, IL6, and VEGF, in HeLa CCL2 cells treated with MO-2097 under hypoxic conditions.

Figure 11 shows the results of confirming the anticancer activity of MO-2097 in a 3D spheroid model of HCT116 cells using a Zeiss microscope.

Figure 12 shows the average values of the spheroid areas confirmed in Figure 10 organized by date.

Figure 13 shows the results of treating HCT116 spheroids with DMSO or 25 and 50 μM MO-2097, culturing the spheroids for 2 more days, fixing them in 4% paraformaldehyde, and then staining the samples with anti-cleaved caspase 3 antibody (green) and Tubulin antibody (red) after cryosectioning.

Figure 14 is a mouse image showing the results of a xenograft analysis when both vehicle and MO-2097 were injected into mice with tumors.

Figure 15 is a graph showing the mouse body weight of Figure 14.

Figure 16 is a graph showing the mouse tumor volume of Figure 14.

Figure 17 is a graph showing the relative tumor weight of the mouse in Figure 14.

Figure 18 shows the results of additional tumor resections following in vivo xenograft studies and staining with anti-HIF-1α antibody (green).

Figure 19 shows the results of additional tumor resections following in vivo xenograft studies and staining with anti-cleaved caspase 3 antibody (green).

Figure 20 shows the results of microscopic observation of the growth inhibitory effect on organoids 1, 2, 3, and 4, respectively, after treatment with MO-2097.

Figure 21 is a graph showing the relative values for the growth inhibition effect on organoids 1, 2, 3, and 4 respectively after treatment with MO-2097.

Figure 22 shows the results of lysing organoids 1 and 2 in sample buffer and then blotting with the indicated antibodies.

Figure 23 shows the results of harvesting organoid 2 for qPCR and analyzing each primer set of the target gene.

Figure 24 shows the results of immunostaining with the indicated antibodies after treating organoid 1 with 25 μM MO-2097 for 24 hours, and observing with a fluorescence microscope (Carl Zeiss).

Figure 25 shows the results of the MTT assay for colon cancer cells (DLD-1) of MO-2097.

Figure 26 shows the results of the MTT assay for sorafenib on colon cancer cells (DLD-1).

Figure 27 shows the results of assaying cell migration with MO-2097 at various concentrations and at various time points.

Figure 28 shows the results of assaying cell migration with Sorafenib at various concentrations and over various time periods.

Figure 29 shows a graph showing the extent to which the space where cells fell off is filled according to the concentration of MO-2097 and Sorafenib.

Figure 30 shows the results of a migration analysis of MO-2097 using crystal violet staining.

Figure 31 shows the numerical results of the transport analysis of MO-2097 and Sorafenib at various concentrations.

Figure 32 shows the results of a mobility analysis using DAPI staining at various concentrations of MO-2097 and Sorafenib.

Figure 33 shows the results of Western blotting of MO-2097 and Sorafenib.

Figures 34 to 36 show the results of MTT analysis of compounds 1a-1t of the examples on cervical cancer cells (HeLa CCL2).

Figure 37 shows the results of assaying cell migration of compounds 1a-1t of the example.

Figure 38 summarizes the effects of compounds 1a-1t of the examples confirmed in Figures 34 to 37.

Hereinafter, the present invention will be described in more detail through examples. These examples are intended to explain the present invention more specifically, and the scope of the present invention is not limited to these examples.

¹ H-NMR and ¹³ C-NMR of compounds prepared according to one embodiment of the present invention were measured using a Varian (400 MHz) spectrometer (Varian Medical Systems, Inc., Palo Alto, CA, USA), respectively. ¹ H-NMR data are reported in terms of peak multiplicity as s for a singlet, d for a doublet, dd for a doublet of doublets, t for a triplet, q for a quartet, brs for a broad singlet, and m for a multiplet. In ¹³ C-NMR, chemical shift values are reported in δ (ppm), and coupling constants ( J ) in Hz.

The mass spectrum of the compound prepared according to one embodiment of the present invention was recorded using high-resolution mass spectrometry (HRMS, ESI-MS) obtained on a G2 QTOF mass spectrometer (Waters Corp, Milford, USA). The product was purified by column or flash chromatography (Biotage, Sweden) using silica gel 60 (230-400 mesh Kieselgel 60).

The purity of the compounds prepared according to one embodiment of the present invention was confirmed by reverse phase high pressure liquid chromatography (RP-HPLC) performed on a Waters Corp. HPLC system equipped with an ultraviolet (UV) detector set to 254 nm. The mobile phases used were (A) H ₂ O containing 0.05% trifluoroacetic acid and (B) CH ₃ CN. HPLC was performed using a YMC Hydrosphere C18 (HS-302) column (particle size 5 μm, pore size 12 nm), 4.6 mm in diameter x 150 mm in size, at a flow rate of 1.0 mL/min. The compound purity was assessed using a gradient from 25% B to 100% B in 30 min (Method A) or from 5% B to 100% B in 30 min (Method B). The purity of all compounds evaluated biologically was ≥ 95% by Method A or Method B.

<Experimental example>

Experimental example 1. Experimental process of Suzuki coupling.

Pd(dppf)Cl2·DCM (5 mol%) was added to a solution of degassed 2-bromo-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (compound 4 of the following reaction scheme, 1.0 eq), boronic ester/acid (0.8 eq), and K3PO4 (2.5 eq) in DMSO:H2O (4:1). The reaction was heated at 70 °C for 1 h. The reaction was cooled to room temperature, filtered through Celite, and partitioned between EtOAc and water. The organic layer was washed with brine, dried over sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column chromatography to give the pure product.

Experimental Example 2. Experimental Process of Ester Hydrolysis

LiOH.H2O (5.0 eq) was added to a solution of the ester (1.0 eq) in THF:EtOH:H2O (2:2:1) at 0°C. The resulting mixture was stirred for 3 h and slowly warmed to room temperature. The reaction mixture was distilled to dryness and acidified with 1.5 N HCl. The precipitated solid was filtered and dried under vacuum.

Experimental Example 3. Experimental Process of Amide Coupling

A solution of propylphosphonic anhydride (T3P) (≥50 wt % in EtOAc) (2.0 equiv) was added to a solution of carboxylic acid (1.0 equiv), amine (1.5 equiv) and TEA (3.0 equiv) in DCM, and the reaction mixture was stirred at room temperature overnight. The reaction was treated with water and extracted with DCM. The combined organic layers were washed with brine, dried over sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column chromatography to give the pure product.

Experimental Example 4. Experimental Process of Miyaura Borilization

Pd(dppf)Cl2·DCM (5 mol%) was added to a degassed solution of aryl bromide (1 equiv.), bis(pinacolato)diboron (1.2 equiv.), and KOAc (2.0 equiv.) in 1,4-dioxane (Method A) or DMF (Method B). The reaction mixture was heated to 90 °C for 5 h. The reaction was cooled to room temperature, filtered through Celite, and concentrated in vacuo. The residue was purified by silica gel column chromatography to give the pure product (Method A) or the reaction was cooled to room temperature, treated with water, and extracted with EtOAc. The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column chromatography to give the pure product (Method A).

<Example>

Example 1. Synthesis of 2-bromo-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran

(1) Preparation of 2-bromo-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (4 in the reaction scheme) from 5-allyl-2-bromobenzofuran-6-ol (2 in the reaction scheme)

[Reaction Formula 1]

First, 2-bromo-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (4 in the reaction scheme) was prepared from 5-allyl-2-bromobenzofuran-6-ol (2 in the reaction scheme) with reference to the above reaction scheme 1. Specifically, 5-allyl-2-bromobenzofuran-6-ol (2 in the reaction scheme) was mixed with allyl chloride, NaI, Cs ₂ CO ₃ and acetone and alkylation reaction was performed at 60° C. for 2 hours to obtain 5-allyl-2-bromo-6-[(2-methylallyl)oxy]benzofuran (3 in the reaction scheme) with a yield of about 80% (step a).

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.23 (s, 1H), 6.94 (s, 1H), 6.59 (s, 1H), 6.05 - 5.98 (m, 1H), 5.13 (s, 1H, merged), 5.07 (dd, J = 1.6 Hz, 16.8 Hz, 1H), 5.05 (dd, J = 1.6 Hz, 11.6 Hz, 1H), 5.00(s, 1H, merged), 4.44 (s, 2H), 3.46 (d, J = 6.4 Hz, 2H), 1.55(s, 2H); ¹³ C NMR (100 MHz, CDCl ₃ ) δ 155.22, 154.36, 140.64, 137.10, 125.57, 125.53, 121.41, 120.07, 115.51, 112.59, 107.88, 94.84, 72.01, 34.63, 19.48; HRMS (ESI): [M+H] ⁺ C ₁₅ H ₁₆ BrO ₂ calcd 307.0334, found 307.0340.

Thereafter, the above 5-allyl-2-bromo-6-[(2-methylallyl)oxy]benzofuran (3 in the reaction scheme) was subjected to a ring-closing metathesis (RCM) reaction using Grubb's ^2nd generation catalyst at 60°C for 2 hours to produce 2-bromo-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (4 in the reaction scheme) in a yield of about 88% (step d).

^1H NMR (400 MHz, CDCl ₃ ) δ 7.18 (s 1H),7.17 (s, 1H), 6.61(s, 1H), 5.39 (t, J = 4.8 Hz, 1H), (m, 1H), 4.45 (s, 2H), 3.44 (d, J = 4.8 Hz, 2H), 1.52(s, 3H); ¹³ C NMR (100 MHz, CDCl ₃ ) δ 155.95, 154.80, 133.45, 133.37, 127.39, 124.74, 120.78, 118.17, 107.98, 104.21, 75.11, 30.71, 19.86; HRMS (ESI): [M+H] ⁺ C ₁₃ H ₁₂ BrO ₂ calcd 279.0021, found 279.0013.

(2) Preparation of 2-bromo-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (4 in the reaction scheme) from 2,4-dihydroxy-5-(3-methylbut-1-en-1-yl)benzaldehyde (5 in the reaction scheme)

The above 2-bromo-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (4 in the reaction scheme) can also be produced by a reaction as in the reaction scheme 2 below in addition to the above method (1).

[Reaction Formula 2]

Specifically, 2,4-dihydroxy-5-(3-methylbut-1-en-1-yl)benzaldehyde (5 in the reaction scheme), a known aldehyde that can be easily obtained from 2,4-dihydroxybenzaldehyde, was prepared. Thereafter, the above 2,4-dihydroxy-5-(3-methylbut-1-en-1-yl)benzaldehyde (5 in the reaction scheme) was mixed with 2-methylprop-2-en-1-ol, PPh3, DIAD and toluene: _CH3CN (2:1), and alkylation was performed at 0°C to room temperature for 2 hours to obtain 2-hydroxy-4-[(2-methylallyl)oxy]-5-(3-methylbut-2-en-1-yl)benzaldehyde (6 in the reaction scheme) in a yield of about 63% (step b).

^1H NMR (400 MHz, CDCl ₃ ) δ 11.42 (s, 1H), 9.68 (s, 1H), 7.22 (s, 1H), 6.37 (s, 1H), 5.28 (t, J = 6.4 Hz, 1H), 5.26 (s, 1H), 5.10 (s, 1H), 4.28 (s, 2H), 3.28 (d, J = 7.2 Hz, 2H), 1.83 (s, 3H), 1.76 (s, 3H), 1.69 (s, 3H); ¹³ C NMR (100 MHz, CDCl ₃ ) δ 194.45, 163.61, 163.04, 139.69, 133.37, 133.31, 122.91, 121.69, 114.40, 113.18, 99.53, 71.96, 27.55, 25.76, 19.30, 17.74; HRMS (ESI): [M+H] ⁺ C ₁₆ H ₂₁ O ₃ calcd 261.1491, found 261.1498.

Finally, 2-hydroxy-4-[(2-methylallyl)oxy]-5-(3-methylbut-2-en-1-yl)benzaldehyde (Scheme 6) was subjected to one-pot cyclisation followed by metathesis at 60°C for 2 h using Grubb's 2 ^nd generation catalyst to give 2-bromo-6-((2-methylallyl)oxy)-5-(4-methylbut-2-en-1-yl)benzofuran (Scheme 7) (step d).

^1H NMR (400 MHz, CDCl ₃ ) δ 7.21 (s, 1H), 6.92 (s, 1H), 6.58 (s, 1H), 5.35 (t, J = 6.4 Hz, 1H), 5.12 (s, 1H), 5.00 (s, 1H), 4.45 (s, 2H), 3.40 (d, J = 7.2 Hz, 2H), 1.84 (s, 3H), 1.74 (s, 3H), 1.70 (s, 3H); ¹³ C NMR (100 MHz, CDCl ₃ ) δ 155.03, 154.46, 140.72, 132.58, 127.01, 125.34, 122.70, 121.35, 119.54, 112.61, 107.88, 94.76, 72.03, 28.75, 25.80, 19.42, 17.76.

Thereafter, 2-bromo-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (4) was prepared from 2-bromo-6-((2-methylallyl)oxy)-5-(4-methylbut-2-en-1-yl)benzofuran (7 in the reaction scheme) (step d). At this time, 2-bromo-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (4 in the reaction scheme) was prepared from 2-bromo-6-((2-methylallyl)oxy)-5-(4-methylbut-2-en-1-yl)benzofuran (7 in the reaction scheme) in a yield of about 84%, and the overall yield was 31%.

Example 2. Synthesis of MO-2097 and derivatives (1a-1t)

Through the Suzuki reaction of the above Experimental Example 1, MO-2097 and derivatives (1a-1t) were synthesized from 2-bromo-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (4 in the reaction scheme) as shown in the following reaction scheme 3.

[Reaction Formula 3]

^a Reagents and conditions: (a) Boronic acid ester/acid (8a-8t), K ₃ PO ₄ , Pd(dppf)Cl ₂ ·DCM, DMSO:H ₂ O (4:1), 70°C, 1 hour, 15-75%

(1) Synthesis of 5-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)benzene-1,3-diol (MO-2097) (1a in the reaction scheme)

Compound 1a of the reaction scheme was prepared from 2-bromo-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (4 of the reaction scheme) and 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzene-1,3-diol (8a of the reaction scheme) according to the Suzuki coupling procedure of Experimental Example 1. The residue was purified by silica gel column chromatography (0-35% EtOAc in hexane) to give 1a as a white solid (0.40 g, 70%). mp: 248-250 °C; ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 9.48 (brs, 2H), 7.35 (s, 1H), 7.31 (s, 1H), 7.14 (s, 1H), 6.72 (d, J = 2.0 Hz, 2H), 6.24 (t, J = 2.0 Hz, 1H), 5.63 (t, J = 2.8 Hz, 1H), 4.43 (s, 2H), 3.40 (d, J = 2.8 Hz, 2H), 1.49 (s, 3H); ¹³ C NMR (100 MHz, DMSO- d ₆ ) δ 159.28, 156.25, 156.20, 153.59, 133.92, 133.03, 131.77, 125.17, 120.89, 119.60, 104.57, 103.48, 103.02, 101.83, 74.81, 30.41, 19.95; HRMS (ESI): [M+H] ⁺ C ₁₉ H ₁₇ O ₄ calcd 309.1127, found 309.1132; purity > 99.9% (as determined by RP-HPLC, method A, t _R =15.91 min).

(2) Synthesis of 4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)phenol (1b in the reaction scheme)

Compound 1b of the reaction scheme was prepared from 2-bromo-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (4 of the reaction scheme) and 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (8b of the reaction scheme) according to the Suzuki coupling procedure of Experimental Example 1. The residue was purified by silica gel column chromatography (0-15% EtOAc in hexane) to give 1b as a white solid (20.0 mg, 55%). mp: 171.5-173.5 °C; ^1H NMR (400 MHz, Methanol- d ₄ ) δ 7.88 (d, J = 8.8 Hz, 2H), 7.22 (s, 1H), 7.16 (s, 1H), 6.87-6.83 (m, 3H), 5.65 (t, J = 2.8 Hz, 1H), 4.45 (s, 2H), 3.42 (d, J = 2.8 Hz, 2H), 1.53 (s, 3H); ¹³ C NMR (100 MHz, Methanol- d ₄ ) δ 157.20, 156.09, 155.03, 153.08, 132.70, 131.87, 125.18, 125.06, 121.54, 119.98, 117.71, 114.65, 102.66, 97.82, 74.08, 29.59, 17.96; HRMS (ESI): [M+H] ⁺ C ₁₉ H ₁₇ O ₃ calcd 293.1178, found 293.1173.

(3) Synthesis of 4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)benzene-1,2-diol (1c in the reaction scheme)

Compound 1c of the reaction scheme was prepared from 2-bromo-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (4 of the reaction scheme) and (3,4-dihydroxyphenyl)boronic acid (8c of the reaction scheme) according to the Suzuki coupling procedure of Experimental Example 1. The residue was purified by silica gel column chromatography (0-2% MeOH in DCM) to give 1c as a white solid (29.0 mg, 43%). mp: 193-195 °C; ^1H NMR (400 MHz, Methanol _-d4 ) δ 7.25 (d, J = 2.0 Hz, 1H), 7.21 (s, 1H), 7.19 (dd, J = 8.4, 2.0 Hz, 1H), 7.15 (s, 1H), 6.82 (d, J = 8.4 Hz, 1H) ), 6.80 (s, 1H), 5.65 (t, J = 2.8 Hz, 1H), 4.44 (s, 2H), 3.43 (d, J = 2.8 Hz, 2H), 1.52 (s, 3H); ¹³ C NMR (101 MHz, Methanol-d ₄ ) δ 158.20, 157.03, 155.08, 147.30, 146.66, 134.73, 133.87, 127.07, 123.99, 121.99, 119.67, 117.88, 116.65, 112.80, 104.63, 99.90, 76.10, 31.59, 19.96; HRMS (ESI): [M+H] ⁺ C ₁₉ H ₁₇ O ₄ calcd 309.1127, found 309.1157; purity > 95.62% (as determined by RP-HPLC, method A, t _R =17.17 min).

(4) Synthesis of 3-methoxy-5-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)phenol (1d in the reaction scheme)

Compound 1d of the reaction scheme was prepared from 2-bromo-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (4 of the reaction scheme) and 3-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (8d of the reaction scheme) according to the Suzuki coupling procedure of Experimental Example 1. The residue was purified by silica gel column chromatography (0-30% EtOAc in hexane) to give 1d as a white solid (20.0 mg, 28%). mp: 172.5-174.5 °C; ¹ H NMR (400 MHz, Methanol- d ₄ ) δ 7.26 (s, 1H), 7.19 (s, 1H), 7.00 (s, 1H), 6.89 (s, 1H), 6.87 (s, 1H), 6.36 (s, 1H), 5.65 (t, J = 2.8 Hz, 1H), 4.45 (s, 2H), 3.81 (s, 3H), 3.43 (d, J = 2.8 Hz, 2H), 1.52 (s, 3H); ¹³ C NMR (100 MHz, Methanol- d ₄ ) δ 160.73, 158.09, 155.69, 155.53, 153.31, 132.74, 132.22, 131.55, 124.63, 119.93, 118.14, 103.23, 102.84, 100.63, 100.58, 100.41, 74.12, 53.78, 29.57, 17.95; HRMS (ESI): [M+H] ⁺ C ₂₀ H ₁₉ O ₄ calcd 323.1283, found 323.1273; purity > 98.68% (as determined by RP-HPLC, method A, t _R = 20.08 min).

(5) Synthesis of methyl 4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)benzoate (1e in the reaction scheme)

Compound 1e of the reaction scheme was prepared from 2-bromo-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (4 of the reaction scheme) and (4-(methoxycarbonyl)phenyl)boronic acid (8e of the reaction scheme) according to the Suzuki coupling procedure of Experimental Example 1. The residue was purified by silica gel column chromatography (0-15% EtOAc in hexane) to give 1e as a white solid (110.0 mg, 76%). mp: 192-194 °C; ^1H NMR (400 MHz, DMSO _-d6 ) δ 8.06 (d, J = 8.4 Hz, 2H), 8.01 (d, J = 8.4 Hz, 2H), 7.56 (s, 1H), 7.45 (s, 1H), 7.37 (s, 1H), 5.64 (t, J = 2.8 Hz, 1H), 4.45 (s, 2H), 3.88 (s, 3H), 3.43 (d, J = 2.8 Hz, 2H), 1.50 (s, 3H); ¹³ C NMR (100 MHz, DMSO- d ₆ ) δ 165.65, 156.50, 153.97, 153.69, 133.88, 133.38, 133.00, 129.80, 128.83, 124.42, 124.28, 120.27, 119.61, 104.26, 104.23, 74.30, 52.11, 29.81, 19.39; HRMS (ESI): [M+H] ⁺ C ₂₁ H ₁₉ O ₄ calcd 335.1283, found 335.1281; purity > 95.03% (as determined by RP-HPLC, method A, t _R = 25.98 min).

(6) Synthesis of 4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)benzoic acid (1f in the reaction scheme)

Compound 1f of the reaction scheme was prepared from methyl 4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)benzoate (1e of the reaction scheme) according to the general procedure for hydrolysis in Experimental Example 2 to give 1f as a white solid (60.0 mg, 78%). ^1H NMR (400 MHz, DMSO _-d6 ) δ 7.94 (d, J = 8.0 Hz, 2H), 7.76 (d, J = 8.0 Hz, 2H), 7.38 (s, 1H), 7.33 (s, 1H), 7.30 (s, 1H), 5.64 (t, J = 2.8 Hz, 1H), 4.44 (s, 2H), 3.42 (d, J = 2.8 Hz, 2H), 1.50 (s, 3H); ¹³ C NMR (100 MHz, DMSO- d ₆ ) δ 168.31, 155.86, 153.37, 140.95, 133.49, 132.64, 129.65, 124.84, 123.22, 120.45, 119.20, 104.16, 101.64, 74.37, 29.97, 19.51; HRMS (ESI): [M+H] ⁺ C ₂₀ H ₁₇ O ₄ calcd 321.1127, found 321.1159.

(7) Synthesis of [4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)phenyl](4-methylpiperazin-1-yl)methanone (1g in the reaction scheme)

Compound 1 g of the reaction scheme was prepared from 4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)benzoic acid (scheme 1f) and 1-methylpiperazine according to the general amide coupling procedure of Experimental Example 3. The residue was purified by silica gel column chromatography (0-10% MeOH in DCM) to give 1 g as a white solid (20.0 mg, 56%). mp: 112-115 °C; ^1H NMR (400 MHz, CDCl ₃ ) δ 7.85 (d, J = 8.4 Hz, 2H ), 7.47 (d, J = 8.4 Hz, 2H ), 7.27 (s, 1H), 7.26 (merged) (s, 1H), 6.99 (s, 1H), 5.67 (t, J = 2.8 Hz, 1H), 4.48 (s, 2H), 3.81 (brs, 2H), 3.48 (s, 4H), 2.50 (brs, 2H), 2.39 (brs, 2H), 2.33 (s, 3H), 1.53 (s, 3H); ¹³ C NMR (100 MHz, CDCl ₃ ) 169.87, 156.55, 155.12, 154.28, 135.19, 133.45, 133.13, 131.87, 127.74, 125.08, 124.55, 120.94, 119.16, 104.44, 102.19, 75.24, 55.25, 54.68, 47.72, 46.01, 42.18, 30.82, 19.88; HRMS (ESI): [M+H] ⁺ C ₂₅ H ₂₇ N ₂ O ₃ calcd 403.2022, found 403.2002; purity > 95.26% (as determined by RP-HPLC, method B, t _R = 19.66 min).

(8) Synthesis of 2-fluoro-4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)phenol (1h in the reaction scheme)

Compound 1h of the reaction scheme was prepared from 2-bromo-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (4 of the reaction scheme) and 2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (8h of the reaction scheme) according to the Suzuki coupling procedure in Experimental Example 3. The residue was purified by silica gel column chromatography (0-15% EtOAc in hexane) to give 1h as a white solid (25.0 mg, 30%). ^1H NMR (400 MHz, CDCl ₃ ) δ 7.55-7.48 (m, 2H), 7.26 (s, 1H), 7.23 (s, 1H), 7.04 (t, J = 8.0 Hz, 1H), 6.79 (s, 1H), 5.66 (brs, 1H), 5.43 (t, J = 2.8 Hz, 1H), 4.48 (s, 2H), 3.46 ( d, J = 2.8 Hz, 2H), 1.53 (s, 3H);HRMS (ESI): [M+H] ⁺ C ₁₉ H ₁₆ O ₃ F calcd 311.1083, found 311.1076.

(9) Synthesis of N,N-dimethyl-4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)benzamide (1i in the reaction scheme)

Compound 1i of the reaction scheme was prepared from 2-bromo-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (4 of the reaction scheme) and 4-(dimethylcarbamoyl)phenylboronic acid (8i of the reaction scheme) according to the Suzuki coupling procedure of Experimental Example 3. The residue was purified by silica gel column chromatography (0-40% EtOAc in hexane) to give 1i as a white solid (23.0 mg, 35%). mp: 162.5-165.5 °C; ^1H NMR (400 MHz, CDCl ₃ ) δ 7.85 (d, J = 8.4 Hz, 2H), 7.49 (d, J = 8.4 Hz, 2H), 7.27 (s, 1H), 7.25 (s, 1H), 6.99 (s, 1H), 5.67 (t, J = 2.8) Hz, 1H), 4.49 (s, 2H), 3.48 (d, J = 2.8 Hz, 2H), 3.13 (s, 3H), 3.03 (s, 3H), 1.64 (s, 3H; ¹³ C NMR (101 MHz, CDCl ₃ ) δ 171.11, 156.48, 155.22, 154.24, 135.77, 133.44, 133.07, 131.65, 127.72, 125.08, 124.42, 120.93, 119.12, 104.41, 102.06, 75.21, 39.57, 35.41, 30.80, 19.87; HRMS (ESI): [M+H] ⁺ C ₂₂ H ₂₂ NO ₃ calcd 348.1600, found 348.1595; Purity> 98.16% (as determined by RP-HPLC, method A, t _R = 19.42 min).

(10) Synthesis of 2-(4-fluorophenyl)-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (1j in the reaction scheme)

Compound 1j of the reaction scheme was prepared from 2-bromo-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (4 of the reaction scheme) and (4-fluorophenyl)boronic acid (8j of the reaction scheme) according to the Suzuki coupling procedure of Experimental Example 3. 1j was obtained as a white solid (25.0 mg, 40%), after being purified by silica gel column chromatography (0-1% EtOAc in hexane). mp: 165.5-165.5 °C; ^1H NMR (400 MHz, CDCl ₃ ) δ 7.81-7.77 (m, 2H), 7.26 (s, 1H), 7.24 (s, 1H), 7.12 (t, J = 8.8 Hz, 1H), 6.85 (s, 1H), 5.66 (t, J = 2.8 Hz, 1H), 4.48 (s, 2H), 3.47 (d, J = 2.8 Hz, 2H), 1.55 (s, 3H); ¹³ C NMR (125 MHz, CDCl ₃ ) δ 156.16, 155.29, 154.06, 133.48, 132.95, 126.91, 126.44, 125.26, 120.98, 118.96, 115.95, 115.77, 104.36, 100.74, 77.30, 77.04, 76.79, 75.23, 30.96, 30.82, 19.91; HRMS (ESI): [M+H] ⁺ C ₁₉ H ₁₆ FO ₂ calcd 295.1134, found 295.1134; Purity> 96.27% (as determined by RP-HPLC, method A, t _R = 26.26 min).

(11) Synthesis of 2-(3,5-difluorophenyl)-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (1k in the reaction scheme)

Compound 1k of the reaction scheme was prepared from 2-bromo-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (4 of the reaction scheme) and (3,5-difluorophenyl)boronic acid (8k of the reaction scheme) according to the Suzuki coupling procedure of Experimental Example 3. The residue was purified by silica gel column chromatography (0-2% EtOAc in hexane) to give 1k as a white solid (40.0 mg, 60%). mp: 125.5-128.5 °C; ^1H NMR (400 MHz, CDCl ₃ ) δ 7.35-7.29 (m, 2H), 7.28 (s, 1H), 7.25 (s, 1H), 6.95 (d, J = 4.4 Hz, 1H), 6.75 (tt, J = 8.8, 2.4 Hz, 1H), 5.66 (t, J = 2.8 Hz, 1H), 4.48 (s, 2H), 3.47 (d, J = 2.8 Hz, 2H), 1.54 (s, 3H); ¹³ C NMR (125 MHz, CDCl ₃ ) δ 164.37, 164.27, 162.40, 162.30, 156.88, 154.23, 153.68, 133.45, 133.39, 124.74, 120.85, 119.35, 107.51, 107.45, 107.34, 107.29, 104.51, 103.57, 103.37, 103.17, 103.06, 75.22, 30.76, 19.86; HRMS (ESI): [M+H] ⁺ C ₁₉ H ₁₂ F ₂ O ₂ calcd 313.1040, found 313.1047; Purity> 95% (as determined by RP-HPLC, method A, t _R = 27.01 min).

(12) Synthesis of [4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)phenyl](piperidin-1-yl)methanone (1l in the scheme)

Compound 1l of the reaction scheme was prepared from 2-bromo-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (4 in the reaction scheme) and piperidin-1-yl(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanone (8l in the reaction scheme) according to the Suzuki coupling procedure in Experimental Example 3. The product was purified by silica gel column chromatography (0-35% EtOAc in hexane) to give 1l as a white solid (40.0 mg, 37%). mp: 168-169 °C; ^1H NMR (400 MHz, CDCl ₃ ) δ 7.84 (d, J = 8.0 Hz, 2H ), 7.46 (d, J = 8.0 Hz, 2H ), 7.27 (s, 1H), 7.01 (s, 1H), 6.98 (s, 1H), 5.67 (t, J = 5.0) Hz, 1H), 4.49 (s, 2H), 3.73 (brs, 2H), 3.48 (d, J = 5.0 Hz, 2H), 3.39 (brs, 2H), 1.69 (brs, 4H), 1.53 (brs, 5H); ¹³ C NMR (125 MHz, CDCl ₃ ) 169.87, 156.55, 155.12, 154.28, 135.19, 133.45, 133.13, 131.87, 127.74, 125.08, 124.55, 120.94, 119.16, 104.44, 102.19, 75.24, 55.25, 54.68, 47.72, 46.01, 42.18, 30.82, 19.88; HRMS (ESI): [M+H] ⁺ C ₂₅ H ₂₆ NO ₃ calcd 388.1913, found 388.1900; Purity> 97.82% (as determined by RP-HPLC, method A, t _R = 23.47 min).

(13) Synthesis of 4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)-N-(1-methylpiperidin-4-yl)benzamide (1m in the reaction scheme)

Compound 1m of the reaction scheme was prepared from 2-bromo-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (4 in the reaction scheme) and N-(1-methylpiperidin-4-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (8m in the reaction scheme) according to the Suzuki coupling procedure in Experimental Example 3. The residue was purified by silica gel column chromatography (0–20% MeOH in DCM) to give 1m as a brown solid (37.0 mg, 32%). mp: 233–235 °C; ^1H NMR (400 MHz, CDCl ₃ ) δ 7.87 (d, J = 8.4 Hz, 2H ), 7.82 (d, J = 8.4 Hz, 2H ), 7.28 (s, 1H), 7.26 (s, 1H), 7.03 (s, 1H), 6.09 (d, J = 7.6) Hz, 1H), 5.66 (t, J = 7.6 Hz, 1H), 4.49 (s, 2H), 4.05 (brs, 1H), 3.48 (d, J = 5.0 Hz, 2H), 2.96 (d, J = 10.0 Hz, 2H), 2.38 (s, 3H), 2.28 (t, J = 11.2 Hz, 2H ), 2.10 (t, J = 12.0 Hz, 2H ), 1.76 - 1.67 (m, 2H), 1.54 (s, 3H); ¹³ C NMR (125 MHz, CDCl ₃ ) 166.31, 156.70, 154.91, 154.37, 133.75, 133.46, 133.40, 133.21, 127.44, 125.03, 124.55, 120.92, 119.24, 104.47, 102.77, 75.24, 54.45, 46.36, 45.83, 31.89, 30.81, 19.88; HRMS (ESI): [M+H] ⁺ C ₂₆ H ₂₉ N ₂ O ₃ calcd 417.2178, found 417.2194; Purity> 98.71% (as determined by RP-HPLC, method B (XXZ Test), t _R = 21.90 min).

(14) Synthesis of N-benzyl-4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)benzamide (1n in the reaction scheme)

Compound 1n of the reaction scheme was prepared from 2-bromo-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (4 in the reaction scheme) and N-benzyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (8n in the reaction scheme) according to the Suzuki coupling procedure in Experimental Example 3. The residue was purified by silica gel column chromatography (0-30% EtOAc in hexane) to give 1n as a white solid (55.0 mg, 37%). mp: 215-217 °C; ^1H NMR (400 MHz, CDCl ₃ ) δ 7.89 - 7.83 (m, 4H), 7.39 - 7.31 (m, 5H), 7.27 (s, 1H), 7.26 (s, 1H), 7.03 (s, 1H), 6.45 (t, J = 5.6 Hz, 1H), 5.65 (t, J = 5.6 Hz, 1H), 4.67 (d, J = 5.6 Hz, 2H), 4.49 (s, 2H), 3.48 (d, J = 5.0 Hz, 2H), 1.54 (s, 3H); ¹³ C NMR (125 MHz, CDCl ₃ ) δ 166.71, 156.70, 154.90, 154.37, 138.11, 133.53, 133.46, 133.21, 128.84, 127.99, 127.70, 127.52, 125.03, 124.57, 120.92, 119.25, 104.47, 102.80, 75.24, 44.22, 30.81, 19.88; HRMS (ESI): [M+H] ⁺ C ₂₇ H ₂₃ NO ₃ calcd 410.1756, found 410.1736; Purity≥99.9% (as determined by RP-HPLC, method A, t _R = 23.14 min).

(15) Synthesis of tert-butyl 4-[4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)benzoyl]piperazine-1-carboxylate (1o in the reaction scheme)

Compound 1o of the reaction scheme was prepared from 2-bromo-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (4 of the reaction scheme) and tert-butyl 4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoyl]piperazine-1-carboxylate (8o of the reaction scheme) according to the Suzuki coupling procedure of Experimental Example 3. The residue was purified by silica gel column chromatography (0 to 25% EtOAc in hexane) to give 1o (80.0 mg, 46%) as a white solid. mp: 158.5-160.5 °C; ^1H NMR (400 MHz, CDCl ₃ ) δ 7.86 (d, J = 8.4 Hz, 2H ), 7.48 (d, J = 8.4 Hz, 2H ), 7.28 (s, 1H), 7.26 (s, 1H), 7.00 (s, 1H), 5.67 (t, J = 7.6) Hz, 1H), 4.49 (s, 2H), 3.74 (brs, 2H), 3.47 (brs, 8H), 1.54 (s, 3H), 1.47 (s, 9H); ¹³ C NMR (125 MHz, CDCl ₃ ) δ 170.16, 156.61, 155.01, 154.57, 154.31, 134.82, 133.47, 133.19, 132.09, 127.77, 125.05, 124.64, 120.93, 119.19, 104.47, 102.33, 80.40, , 75.24, 30.94, 30.81, 28.37, 19.89; HRMS (ESI): [M+H] ⁺ C ₂₉ H ₃₂ N ₂ O ₅ calcd 489.2389, found 489.2379; Purity≥98.14% (as determined by RP-HPLC, method A, t _R 24.10 min).

(16) Synthesis of [4-(4-fluorobenzyl)piperazin-1-yl][4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)phenyl]methanone (1p in the scheme)

Compound 1p of the reaction scheme was prepared from 2-bromo-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (4 of the reaction scheme) and [4-(4-fluorobenzyl)piperazin-1-yl][4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanone (8p of the reaction scheme) according to the Suzuki coupling procedure in Experimental Example 3. The residue was purified by silica gel column chromatography (0-30% EtOAc in hexane) to give 1p as a white solid (90.0 mg, 46%). mp: 168.5-170.5 °C; ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.84 (d, J = 8.4 Hz, 2H), 7.47 (d, J = 8.4 Hz, 2H), 7.31 - 7.26 (m, 4H), 7.05 - 6.97 (m, 3H), 5.67 (t, J = 5.2 Hz, 1H), 4.49 (s, 2H), 3.80 (brs, 2H), 3.51 (s, 2H), 3.48 (d, J = 5.2 Hz, 2H), 2.59 (brs, 2H), 2.39 (brs, 2H)1.59 (s, 3H).; ¹³ C NMR (125 MHz, CDCl ₃ ) δ 169.82, 163.10, 161.11, 156.55, 155.13, 154.27, 135.25, 133.46, 133.14, 131.82, 130.58, 130.52, 127.74, 125.07, 124.53, 120.94, 119.15, 115.26, 115.09, 104.45, 102.17, 75.24, 62.08, 53.23, 52.70, 48.04, 42.19, 30.81, 19.88; HRMS (ESI): [M+H] ⁺ C ₂₉ H ₃₂ N ₂ O ₅ calcd 489.2389, found 489.2379; Purity≥99.9% (as determined by RP-HPLC, method A, t _R = 11.41 min).

(17) Synthesis of [4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)phenyl](piperazin-1-yl)methanone (1q in the reaction scheme)

To a solution of tert-butyl 4-[4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)benzoyl]piperazine-1-carboxylate (compound 1o in the scheme, 1.0 equiv) in DCM at 0 °C was added TFA (10.0 equiv) dropwise. The reaction mixture was slowly warmed to room temperature with stirring for 3 h. The reaction mass was distilled to dryness, basified with saturated bicarbonate solution, and partitioned between EtOAc and water. The organic layer was washed with brine, dried over sodium sulfate, and concentrated in vacuo. The residue was purified by silica gel column chromatography (0-25% MeOH in DCM) to give 1q as a white solid (10.0 mg, 26%). mp: 126.5-129.5 °C; ^1H NMR (400 MHz, CDCl ₃ ) δ 7.85 (d, J = 8.4 Hz, 2H ), 7.47 (d, J = 8.4 Hz, 2H ), 7.28 (s, 1H), 7.26 (s, 1H), 6.99 (s, 1H), 5.67 (t, J = 7.6) Hz, 1H), 4.49 (s, 2H), 3.78 (brs, 2H), 3.47 (s, 2H), 2.97 (brs, 2H), 2.85 (brs, 2H), 1.75 (brs, 2H), 1.54 (s, 3H); ¹³ C NMR (125 MHz, CDCl ₃ ) δ 169.97, 156.55, 155.14, 154.29, 135.27, 133.47, 133.14, 131.82, 127.72, 125.08, 124.58, 120.95, 119.16, 104.45, 102.18, 75.24, 48.91, 46.40, 45.92, 43.36, 30.81, 19.88; HRMS (ESI): [M+H] ⁺ C ₂₄ H ₂₅ N ₂ O ₃ calcd 389.1865, found 389.1861; Purity≥97.02% (as determined by RP-HPLC, method B (XXZ Test), t _R = 20.71 min).

(18) Synthesis of 3-[7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)-N-(1-methylpiperidin-4-yl)benzamide (1r in the reaction scheme)

Compound 1r of the reaction scheme was prepared from 2-bromo-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (4 in the reaction scheme) and N-(1-methylpiperidin-4-yl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (8r in the reaction scheme) according to the Suzuki coupling procedure in Experimental Example 3. The residue was purified by silica gel column chromatography (0–20% MeOH in DCM) to give 1r (70.0 mg, 47%) as a white solid. mp: 225.5–230.5 °C; ^1H NMR (400 MHz, CDCl ₃ ) δ 8.18 (s, 1H), 7.93 (d, J = 8.0 Hz, 1H), 7.68 (d, J = 8.0 Hz, 1H), 7.49 (t, J = 8.0 Hz, 1H), 7.28 (s, 1H), 7.26 (s, 1H), 7.02 (s, 1H), 6.08 (d, J = 7.6 Hz, 1H), 5.67 (t, J = 7.6 Hz, 1H), 4.49 (s, 2H), 4.05 (brs, 1H), 3.48 (d, J = 5.0 Hz, 2H), 2.96 (d, J = 10.0 Hz, 2H ), 2.34 (s, 3H), 2.48 (t, J = 11.2 Hz, 2H), 2.11 (d, J = 12.0 Hz, 2H), 1.76 - 1.67 (m, 2H), 1.54 (s, 3H); ¹³ C NMR (125 MHz, CDCl ₃ ) δ 166.59, 156.52, 155.03, 154.21, 135.37, 133.47, 133.14, 131.08, 129.08, 127.35, 126.38, 125.09, 123.09, 120.95, 119.20, 104.44, 102.10, 54.55, 53.43, 46.61, 46.06, 32.17, 30.81, 19.88.; HRMS (ESI): [M+H] ⁺ C ₂₆ H ₂₉ N ₂ O ₃ calcd 417.2178, found 417.2178; Purity≥97.97% (as determined by RP-HPLC, method B (XXZ Test), t _R = 17.52 min).

(19) Synthesis of 2-methyl-4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)-N-(1-methylpiperidin-4-yl)benzamide (1s in the reaction scheme)

The 1t compound of the reaction scheme was prepared from 2-bromo-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (4 in the reaction scheme) and 2-methyl-N-(1-methylpiperidin-4-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (8s in the reaction scheme) according to the Suzuki coupling procedure in Experimental Example 3. Afterwards, the product was purified by silica gel column chromatography (0-20% MeOH in DCM) to obtain 1s as a white solid (30.0 mg, 20%). mp: 195.5-197.5 °C; ^1H NMR (500 MHz, CDCl ₃ ) δ 7.68 (d, J = 2.4 Hz, 1H), 7.64 (dd, J = 8.0, 2.4 Hz, 1H ), 7.40 (d, J = 8.0 Hz, 1H ), 7.28 (s, 1H), 7.26 (s, 1H), 6.97 (s, 1H), 5.71 (d, J = 7.6 Hz, 1H), 5.67 (t, J = 7.6 Hz, 1H), 4.48 (s, 2H), 4.02 (brs, 1H), 3.48 (d, J = 5.0 Hz, 2H), 2.87 (d, J = 10.0 Hz, 2H ), 2.51 (s, 3H), 2.34 (s, 3H), 2.21 (t, J = 11.2 Hz, 2H), 2.08 (d, J = 12.0 Hz, 2H), 1.66 - 1.58 (m, 2H), 1.54 (s, 3H); ¹³ C NMR (125 MHz, CDCl ₃ ) δ 169.05, 156.48, 155.18, 154.22, 136.65, 135.98, 133.45, 133.09, 131.87, 127.25, 126.95, 125.10, 121.93, 120.95, 119.11, 104.42, 102.13, 77.58, 75.25, 54.46, 46.33, 46.06, 32.14, 30.80, 19.95, 19.89; HRMS (ESI): [M+H] ⁺ C ₂₇ H ₃₁ N ₂ O ₃ calcd 431.2335, found 431.2339; Purity≥97.22% (as determined by RP-HPLC, method B (XXZ Test), t _R = 20.71 min).

(20) Synthesis of 3-methyl-4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)-N-(1-methylpiperidin-4-yl)benzamide (1t in the reaction scheme)

Compound 1t of the reaction scheme was prepared from 2-bromo-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran (4 of the reaction scheme) and 3-methyl-N-(1-methylpiperidin-4-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (8t of the reaction scheme) according to the Suzuki coupling procedure of Experimental Example 3. The residue was purified by silica gel column chromatography (0–20% MeOH in DCM) to give 1t as a white solid (35.0 mg, 22%). mp: 185.5–190.5 °C; ^1H NMR (500 MHz, CDCl ₃ ) δ 7.91 (d, J = 8.0 Hz, 1H ), 7.69 (d, J = 2.4 Hz, 1H), 7.64 (dd, J = 8.0, 2.4 Hz, 1H ), 7.30 (s, 1H), 7.28 (s, 1H), 6.90 (s, 1H), 5.10 (d, J = 7.6 Hz, 1H), 5.67 (t, J = 7.6 Hz, 1H), 4.49 (s, 2H), 4.05 (brs, 1H), 3.49 (d, J = 5.0 Hz, 2H), 2.94 (d, J = 10.0 Hz, 2H ), 2.62 (s, 3H), 2.38 (s, 3H), 2.27 (t, J = 11.2 Hz, 2H), 2.08 (d, J = 12.0 Hz, 2H), 1.75 - 1.70 (m, 2H), 1.53 (s, 3H); ¹³ C NMR (125 MHz, CDCl ₃ ) δ 166.49, 156.64, 154.57, 153.72, 135.64, 133.62, 133.45, 133.08, 132.78, 130.03, 127.72, 125.06, 124.36, 120.96, 119.21, 106.27, 104.34, 77.59, 75.26, 54.50, 46.33, 45.88, 31.93, 30.80, 22.24, 19.89; HRMS (ESI): [M+H] ⁺ C ₂₇ H ₃₁ N ₂ O ₃ calcd 431.2335, found 431.2332; Purity≥99.43% (as determined by RP-HPLC, method B (XXZ Test), t _R = 21.37 min).

Example 3. Synthesis of boronic acid ester (8l-8t)

Among the boronic acid esters/acids (8a-8t) required for the preparation of compounds 1a-1t of the above Example 3, boronic acid esters (8l-t) that are not commercially available were prepared as shown in Scheme 4. Compounds 8l-8t were obtained through two steps of initial amide coupling from the corresponding bromo acids followed by Miyaura borylation.

[Reaction Formula 4]

^a Reagents and conditions: (a) amine, T ₃ P, TEA, DCM, room temperature, 16 h, 97-65%; (b) bis(pinacolato)diboron, KOAc, Pd(dppf)Cl ₂ ·DCM, 1,4-dioxane or MF, 100 °C 16 h, 83-43%.

(1) Synthesis of piperidin-1-yl(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanone (8l in the scheme)

The above compound was prepared from (4-bromophenyl)(piperidin-1-yl)methanone (10l in the scheme) according to the Miyaura borylation procedure method A of Experimental Example 4. It was then purified by silica gel column chromatography (0-35% EtOAc in n-hexane) to obtain 8l as a white solid (0.40 g, 75%). ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.83 (d, J = 8.0 Hz, 2H), 7.37 (d, J = 8.0 Hz, 2H), 3.70 (brs, 2H), 3.29 (brs, 2H), 1.66 (brs, 4H), 1.48 (brs, 2H), 1.35 (s, 12H); HRMS (ESI): [M+H] ⁺ C ₁₈ H ₂₇ NO ₃ B calcd 316.2084, found 316.2071.

(2) Synthesis of N-(1-methylpiperidin-4-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (8m in the reaction scheme)

The above compound was prepared from 4-bromo-N-(1-methylpiperidin-4-yl)benzamide (10m in the scheme) according to the Miyaura borylation procedure A of Experimental Example 4. It was then purified by silica gel column chromatography (0-20% methanol in DCM) to give 8m as a white solid (0.16 g, 47%). ^1H NMR (400 MHz, CDCl ₃ ) δ 7.86 (d, J = 8.0 Hz, 2H), 7.73 (d, J = 8.0 Hz, 2H), 6.00 (d, J = 7.2 Hz, 1H), 4.10 - 3.99 (m, 1H), 2.85 (d, J = 11.6 Hz, 2H), 2.31 (s, 3H), 2.16 (t, J = 11.2 Hz, 2H), 2.04 (d, J = 11.6 Hz, 2H), 1.61 - 1.56 (m, 2H), 1.36 (s, 12H); HRMS (ESI): [M+H] ⁺ C ₁₉ H ₃₀ N ₂ O ₃ B calcd 345.2349, found 345.2377.

(3) Synthesis of N-benzyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (8n in the reaction scheme)

The above compound was prepared from N-benzyl-4-bromobenzamide (10n in the scheme) according to the Miyaura borylation procedure Method A in Experimental Example 4. It was then purified by silica gel column chromatography (0-20% EtOAc in n-hexane) to give 8n as a brown solid (0.30 g, 83%). ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.86 (d, J = 8.0 Hz, 2H), 7.77 (d, J = 8.0 Hz, 2H), 7.37 - 7.26 (m, 5H), 6.45 (t, J = 6.4 Hz, 1H), 4.65 (d, J = 6.4 Hz, 2H ), 1.35 (s, 9H); HRMS (ESI): [M+H] ⁺ C ₂₀ H ₂₅ NO ₃ B calcd 338.1927, found 338.1912.

(4) Turt - Synthesis of butyl 4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoyl]piperazine-1-carboxylate (8o in the scheme)

The above compound was prepared from (4-bromophenyl)(piperidin-1-yl)methanone (10o in the scheme) according to the Miyaura borylation procedure Method A of Experimental Example 4. It was then purified by silica gel column chromatography (0-30% EtOAc in n-hexane) to give 8o as a white solid (0.14 g, 41%). ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.85 (d, J = 8.4 Hz, 2H), 7.38 (d, J = 8.4 Hz, 2H), 3.74 (brs, 2H), 3.51 (brs, 2H), 3.35 (brs, 4H), 1.46 (s, 9H), 1.35 (s, 12H); HRMS (ESI): [M+H] ⁺ C ₂₂ H ₃₄ N ₂ O ₅ B calcd 417.2561, found 417.2560.

(5) Synthesis of [4-(4-fluorobenzyl)piperazin-1-yl][4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanone (8p in the scheme)

The above compound was prepared from (4-bromophenyl)[4-(4-fluorobenzyl)piperazin-1-yl]methanone (10p in the scheme) according to the Miyaura borylation procedure method A of Experimental Example 4. It was then purified by silica gel column chromatography (0-60% EtOAc in n-hexane) to give 8p as a white solid (0.2 g, 60%). ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.83 (d, J = 8.0 Hz, 2H), 7.37 (d, J = 8.0 Hz, 2H), 7.27 (d, J = 8.0 Hz, 2H), 7.00 (t, J = 8.4 Hz, 2H), 3.78 (brs, 2H), 3.48 (s, 2H), 3.37 (brs, 2H), 2.51 (s, 2H), 2.33 (s, 2H) 1.35 (s, 12H); HRMS (ESI): [M+H] ⁺ C ₂₄ H ₃₁ BFN ₂ O ₃ calcd 425.2412, found 425.2411.

(6) Synthesis of N-(1-methylpiperidin-4-yl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (8r in the reaction scheme)

The above compound was prepared from 3-bromo-N-(1-methylpiperidin-4-yl)benzamide (10r in the scheme) according to the Miyaura borylation procedure method B of Experimental Example 4. It was then purified by silica gel column chromatography (0-20% methanol in DCM) to give 8r as a brown solid (0.25 g, 43%). ^1H NMR (400 MHz, CDCl ₃ ) δ 8.05 (s, 1H), 7.95 (d, J = 8.0 Hz, 1H), 7.91 (d, J = 8.0 Hz, 1H), 7.44 (t, J = 8.0 Hz, 1H), 6.19 (d, J = 7.2 Hz, 1H), 4.10 - 3.99 (m, 1H), 3.00 (d, J = 11.6 Hz, 2H), 2.37 (s, 3H), 2.27 (t, J = 11.2 Hz, 2H), 2.05 (d, J = 11.6 Hz, 2H), 1.77 - 1.70 (m, 2H), 1.36 (s, 12H); HRMS (ESI): [M+H] ⁺ C ₁₉ H ₃₀ N ₂ O ₃ B calcd 345.2349, found 345.2340.

(7) Synthesis of 2-methyl-N-(1-methylpiperidin-4-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (8s in the reaction scheme)

The above compound was prepared from 4-bromo-2-methyl-N-(1-methylpiperidin-4-yl)benzamide (10s in the scheme) according to the Miyaura borylation procedure B of Experimental Example 4. It was then purified by silica gel column chromatography (0-20% methanol in DCM) to give 8s as a brown solid (0.35 g, 45%). ^1H NMR (400 MHz, CDCl ₃ ) δ 7.66 (s, 1H), 7.34 (d, J = 8.0 Hz, 1H), 7.21 (d, J = 8.0 Hz, 1H), 5.65 (d, J = 7.5 Hz, 1H), 4.05 - 3.95 (m, 1H), 2.85 (d, J = 11.5 Hz, 2H), 2.41 (s, 3H), 2.34 (s, 3H), 2.22 (t, J = 11.5 Hz, 2H), 2.05 (d, J = 11.5 Hz, 2H), 1.65 - 1.55 (m, 2H) 1.35 (s, 12H); HRMS (ESI): [M+H] ⁺ C ₂₀ H ₃₂ N ₂ O ₃ B calcd 359.2506, found 359.2508.

(8) Synthesis of methyl-N-(1-methylpiperidin-4-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide (8t in the reaction scheme)

The above compound was prepared from 4-bromo-3-methyl-N-(1-methylpiperidin-4-yl)benzamide (10t in the scheme) according to the Miyaura borylation procedure, method B of Experimental Example 4. It was then purified by silica gel column chromatography (0-20% methanol in DCM) to give 8t as a brown solid (0.30 g, 45%). ^1H NMR (400 MHz, CDCl ₃ ) δ 7.81 (d, J = 8.0 Hz, 1H), 7.55 (s, 1H), 7.51 (d, J = 8.0 Hz, 1H), 6.37 (d, J = 7.6 Hz, 1H), 4.27 - 4.00 (m, 1H), 3.47 (d, J = 11.4 Hz, 2H), 2.77 (t, J = 11.4 Hz, 2H), 2.71 (s, 3H), 2.58 (s, 3H), 2.30 (t, J = 11.4 Hz, 2H), 2.08 (d, J = 11.5 Hz, 2H), 1.36 (2, 12H); HRMS (ESI): [M+H] ⁺ C ₂₀ H ₃₂ N ₂ O ₃ B calcd 359.2506, found 359.2504

(9) Synthesis of (4-bromophenyl)(piperidin-1-yl)methanone (10l in the scheme)

The above compound was prepared from 4-bromobenzoic acid (scheme 10a) and piperidine according to the general procedure for amide coupling in Experimental Example 4. It was then purified by silica gel column chromatography (0-35% EtOAc in n-hexane) to give 10 l as a white solid (0.65 g, 97%). ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.54 (d, J = 8.0 Hz, 2H), 7.27 (d, J = 8.0 Hz, 2H), 3.69 (brs, 2H), 3.32 (brs, 2H), 1.68 (brs, 4H), 1.51 (brs, 2H); HRMS (ESI): [M+H] ⁺ C ₁₂ H ₁₅ NOBr calcd 268.0337, found 268.0318.

(10) Synthesis of 4-bromo-N-(1-methylpiperidin-4-yl)benzamide (10m in the reaction scheme)

The above compound was prepared from 4-bromobenzoic acid (10a in Scheme 1) and 1-methylpiperidin-4-amine according to the general procedure for amide coupling in Experimental Example 4. It was then purified by silica gel column chromatography (0-20% methanol in DCM) to give 10m as a white solid (0.55 g, 75%). ^1H NMR (400 MHz, CDCl ₃ ) δ 7.63 (d, J = 8.1 Hz, 2H), 7.57 (d, J = 8.4 Hz, 2H), 6.02 (d, J = 7.6 Hz, 1H) ), 4.05 - 3.95 (m, 1H), 2.88 (d, J = 9.6 Hz, 2H), 2.33 (s, 3H), 2.19 (t, J = 11.2 Hz, 2H), 2.04 (d, J = 11.2 Hz, 2H), 1.64 (q, J = 9.6 Hz, 2H); HRMS (ESI): [M+H] ⁺ C ₁₃ H ₁₈ N ₂ OBr calcd 297.0603, found 297.0588.

(11) Synthesis of N-benzyl-4-bromobenzamide (10n in the reaction scheme)

The above compound was prepared from 4-bromobenzoic acid (10a in Scheme 4) and benzylamine according to the general procedure for amide coupling in Experimental Example 4. It was then purified by silica gel column chromatography (0-30% EtOAc in n-hexane) to give 10n as a white solid (0.65 g, 90%). ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.66 (d, J = 8.4 Hz, 2H), 7.56 (d, J = 8.4 Hz, 2H), 7.39 - 7.26 (m, 5H), 6.38 (t, J = 6.4 Hz, 1H), 4.63 (d, J = 6.4 Hz, 2H ), 1.35 (s, 9H); HRMS (ESI): [M+H] ⁺ C ₁₄ H ₁₃ NOBr calcd 290.0181, found 290.0179.

(12) Turt -Synthesis of butyl 4-(4-bromobenzoyl)piperazine-1-carboxylate (10o in the reaction scheme)

The compound was prepared from 4-bromobenzoic acid (scheme 10a) and tert-butyl piperazine-1-carboxylate according to the general procedure for amide coupling in Experimental Example 4. It was then purified by silica gel column chromatography (0-35% EtOAc in n-hexane) to give 10o as a white solid (0.86 g, 94%). ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.56 (d, J = 8.4 Hz, 2H), 7.28 (d, J = 8.4 Hz, 2H), 3.72 (brs, 2H), 3.50 (brs, 2H), 3.40 (brs, 4H), 1.47 (s, 9H); HRMS (ESI): [M+H] ⁺ C ₁₆ H ₂₂ N ₂ O ₃ Br calcd 369.0814, found 369.0784.

(13) Synthesis of (4-bromophenyl)[4-(4-fluorobenzyl)piperazin-1-yl]methanone (10p of the reaction scheme)

The compound was prepared from 4-bromobenzoic acid (scheme 10a) and 1-(4-fluorobenzyl)piperazine according to the general procedure for amide coupling in Experimental Example 4. It was then purified by silica gel column chromatography (0-2% methanol in DCM) to give 10p as a white solid (0.75 g, 80%). ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.53 (d, J = 8.4 Hz, 2H), 7.29 - 7.25 (m, 4H), 7.00 (t, J = 8.4 Hz, 2H), 3.76 (brs, 2H), 3.49 (s, 2H), 3.40 (brs, 2H), 2.51 (s, 2H), 2.36 (s, 2H); HRMS (ESI): [M+H] ⁺ C ₁₈ H ₁₉ BrFN ₂ O calcd 377.0665, found 377.0662.

(14) Synthesis of 3-bromo-N-(1-methylpiperidin-4-yl)benzamide (10r in the reaction scheme)

The above compound was prepared from 3-bromobenzoic acid (9r in the scheme) and 1-methylpiperidin-4-amine according to the general procedure for amide coupling in Experimental Example 4. It was then purified by silica gel column chromatography (0-20% methanol in DCM) to give 10r as a white solid (0.60 g, 81%). ^1H NMR (500 MHz, CDCl ₃ ) δ 7.88 (t, J = 2.0 Hz, 1H), 7.65 (dt, J = 8.0, 2.0 Hz, 1H), 7.62 (d, J = 8.0, 2.0 Hz, 1H), 7.31 (t, J = 8.0 Hz, 1H), 5.92 (d, J = 7.5 Hz, 1H), 3.99 - 3.95 (m, 1H), 2.82 (d, J = 11.5 Hz, 2H), 2.30 (s, 3H), 2.15 (t, J = 11.5 Hz, 2H), 2.05 (d, J = 11.5 Hz, 2H ) 11.5 Hz, 2H ), 1.65 - 1.55 (m, 2H); HRMS (ESI): [M+H] ⁺ C ₁₃ H ₁₈ N ₂ OBr calcd 297.0603, found 297.0596.

(15) Synthesis of 4-bromo-2-methyl-N-(1-methylpiperidin-4-yl)benzamide (10s in the scheme)

The compound was prepared from 4-bromo-2-methylbenzoic acid (9s in the scheme) and 1-methylpiperidin-4-amine according to the general procedure for amide coupling in Experimental Example 4. It was then purified by silica gel column chromatography (0-20% methanol in DCM) to give 10s as a white solid (0.50 g, 69%). ^1H NMR (500 MHz, CDCl ₃ ) δ 7.38 (s, 1H), 7.34 (d, J = 8.0 Hz, 1H), 7.21 (d, J = 8.0 Hz, 1H), 5.60 (d, J = 7.5 Hz, 1H), 3.99 - 3.95 (m, 1H), 2.82 (d, J = 11.5 Hz, 2H), 2.41 (s, 3H), 2.30 (s, 3H), 2.15 (t, J = 11.5 Hz, 2H), 2.05 (d, J = 11.5 Hz, 2H), 1.65 - 1.55 (m, 2H); HRMS (ESI): [M+H] ⁺ C ₁₄ H ₂₀ N ₂ OBr calcd 311.0750, found 311.0742

(16) Synthesis of 4-bromo-3-methyl-N-(1-methylpiperidin-4-yl)benzamide (10t in the reaction scheme)

The above compound was prepared from 4-bromo-3-methylbenzoic acid (9t in the scheme) and 1-methylpiperidin-4-amine according to the general procedure for amide coupling in Experimental Example 4. It was then purified by silica gel column chromatography (0-20% methanol in DCM) to give 10t as a white solid (0.48 g, 65%). ^1H NMR (500 MHz, CDCl ₃ ) δ 7.63 (d, J = 2.0 Hz, 1H), 7.58 (d, J = 8.0 Hz, 1H), 7.38 (dd, J = 8.0, 2.0 Hz, 1H), 5.93 (d, J = 7.5 Hz, 1H), 4.00 - 3.95 (m, 1H), 2.85 (d, J = 11.5 Hz, 2H), 2.44 (s, 3H), 2.31 (s, 3H), 2.19 (t, J = 11.5 Hz, 2H), 2.05 (d, J = 11.5 Hz, 2H), 1.65 - 1.55 (m, 2H); HRMS (ESI): [M+H] ⁺ C ₁₄ H ₂₀ N ₂ OBr calcd 311.0750, found 311.0757

<Example of an exam>

Test Example 1. Confirmation of HIF-1α protein inhibition and cell death effect

To determine the effect of compound 1a (MO-2097) of the present invention on HIF-1α protein expression, HeLa CCL2 cells were treated with the compound for 24 hours under 1% oxygen conditions and HIF-1α protein levels were measured.

Specifically, HeLa CCL2 cells were seeded in 6-well plates and treated with DMSO control or compound 1a of the present invention (MO-2097) at a concentration of 25 μM for 24 h in hypoxia. HIF-1α protein levels were measured by Western blotting using anti-HIF-1α antibody, whereas hnRNPA2B1 protein levels were measured using anti-hnRNPA2B1 antibody, and the results are shown in Fig. 2 . In Fig. 2 , red stars indicate hnRNPA2B1 protein bands, and black stars are from previous anti-β actin blotting signals.

According to FIG. 2, it can be confirmed that the expression of HIF-1α protein of compound 1a (MO-2097) of the example was particularly reduced.

To determine the effect of compound 1a (MO-2097) of Example 1 on cell growth inhibition, cell death was evaluated by fluorescence-activated cell sorting analysis (FASC) and flow cytometric analysis, and the results are shown in Fig. 3.

Specifically, the FASC analysis was performed as follows. HeLa CCL2 cells were cultured in 12-well culture plates for 18 h. The cells were treated with 25 μM MO-2097 for 24 h under 1% oxygen conditions. The cells were then harvested and stained with propidium iodide (PI) or anti-Annexine V-APC (eBioscience TM Annexin V Apoptosis Detection Kit APC, Invitrogen, Carlsbad, CA, USA) for 30 min to determine the percentage of cells with phosphatidylserine externalization.

Flow cytometry was performed using a flow cytometer (CytoFLEX, Beckman Coulter, Miami, FL, USA). For APC or PI detection, the laser excitation and emission bandpass wavelengths were 650/660 nm or 660/780 nm, respectively. Results were reported as the median of the cell fluorescence intensity distribution obtained by analyzing 10,000 cells in the gate and analyzed using Summit Software (version 6.0, Beckman Coulter, Miami, FL, USA). Internalization scores and mean fluorescence intensity values were calculated for at least 5,000 cells/sample.

The same sample used in the FASC analysis of Fig. 3 above was stained with the Live/Dead TM Viability/Cytotoxicity kit (Life Technologies Corporation) for 30 minutes. The images were then acquired using a Zeiss Axiovert 100M microscope (Carl Zeiss MicroImaging), which are shown in Fig. 4. In Fig. 4, green, red, and blue signals represent live cells, dead cells, and DNA, respectively (Scale bar: 20 μm).

According to FIG. 4, increased apoptosis was observed in compound 1a (MO-2097) of the example. This is consistent with the significant decrease in HIF-1α expression in FIG. 1.

Test Example 2. Confirmation of hnRNPA2B1 binding and HIF-1α reduction effect in hypoxic cells

The binding affinity of hnRNPA2B1 to MO-2097 and Moracin G was measured by ITC analysis. Specifically, His-tagged hnRNPA2B1 protein was purified by Ni ²⁺ bead affinity chromatography, and 50 μM hnRNPA2B1 protein was reacted with 250 nM of each compound at room temperature for 2 h, followed by isothermal titration calorimetry (ITC) analysis.

The above ITC analysis was performed using a MicroCal Auto-iTC200 (Malvern Panalytical, Malvern, UK) at the Korea Basic Science Institute (Ochang, Korea). The binding affinity was measured using 40 μL of 250 μM MO-2097/Moracin G and 200 μL of 50 μM purified hnRNPA2B1 protein in 50 mM Na ₂ HPO ₄ buffer (pH 8.0). For each titration experiment, 2 μL of ligand was injected into the target protein for 4 s at 150 s intervals at 25 °C. In total, 19 injections were performed for each experiment, and the data were analyzed using MicroCal Origin software (Malvern Panalytical).

As a result, the Ka values for the binding affinity of hnRNPA2B1 to MO-2097 and moracin G were 4.13 Х 10 ^-5 /M and 9.32 Х 10 ^-4 /M, respectively. That is, it can be confirmed that MO-2097 has approximately 5 times higher affinity for hnRNPA2B1 than moracin G (Fig. 5).

In addition, to investigate the interaction of MO-2097 and hnRNPA2B1 protein under biochemical conditions, HeLa CCL2 lysate was subjected to biotin pulldown assay. The biotin pulldown assay was performed as follows. HeLa CCL2 cell lysate (1 mg/ml) was incubated with biotin alone or 500 μM biotinylated MO-2097 (Biotin-MO-2097) for 1.5 h. For the competitive control, 1.5 mM MO-2097 was used. Then, 35 μM streptavidin beads were added to each sample and incubated for an additional 1 h. After washing six times, the samples were harvested and Western blotting was used to evaluate the affinity, and the results are shown in Fig. 6. For reference, hnRNPI, hnRNPF/H, and Tubulin antibodies were used as negative binding controls.

At this time, the biotinylated MO-2097 (Biotin-MO-2097) was prepared with reference to the following reaction scheme 5. Specifically, the compound represented by 1a of the reaction scheme (MO-2097) was mixed with ethyl 2-chloroacetate, K ₂ CO ₃ and acetone and alkylated at 60° C. for 16 hours to prepare a compound represented by 11 of the reaction scheme in a yield of about 82% (step a). Thereafter, the compound represented by 11 of the reaction scheme was mixed with LiOH H ₂ O and THF:EtOH:H ₂ O (2:2:1) and hydrolyzed at room temperature for 2 hours to obtain a compound represented by 12 of the reaction scheme in a yield of about 82% (step b). The compound represented by scheme 12 in the above reaction scheme was reacted with a PEG-amine linker under standard amide coupling conditions under one-pot deprotection using TFA to give an amide compound represented by scheme 13 (step c), which was then coupled with (+)-biotin N-hydroxysuccinimide ester in a basic medium to give a compound represented by scheme 14 (Biotin-MO-2097) (step d).

[Reaction Formula 5]

According to Figure 6, Biotin-MO-209 and Biotin-MO-2097 bound well to hnRNPA2B21, but did not bind to other RNA binding proteins, hnRNPI and hnRNPF/H, or the housekeeping protein, Tubulin.

Additionally, the results of seeding HeLa CCL2 cells on poly-L-lysine-coated cover glasses and treating them with DMSO control or 25 and 50 μM MO-2097 for 24 h under hypoxic conditions are shown in Fig. 7. The results of using HCT116 cells instead of HeLa CCL2 cells can be seen in Fig. 8.

At this time, 20% O ₂ conditioned cells were used as a normoxia control, and the samples were stained with anti-HIF-1α antibody (green signal). In Figures 7 and 8, the blue signal represents DNA (Scale bar: 20 μm).

According to Figures 7 and 8, when HeLa CCL2 and HCT116 cells are exposed to hypoxic conditions, HIF-1α protein accumulates, and when treated with the hnRNPA2B1-binding compound MO-2097, it can be confirmed that the protein level of HIF-1α protein decreases.

Test Example 3. Confirmation of the effect of suppressing the expression of HIF-1α protein and HIF-1α target genes

To confirm the inhibitory effect of MO-2097 on HIF-1α protein expression, transcription/translation analysis was performed. Specifically, in vitro transcription/translation HIF-1α analysis was performed in the presence or absence of hnRNPA1B1 protein using 50–100 μM MO-2097, and Western blotting was performed with anti-HIF-1α and anti-hnRNPA2B1 antibodies. Numbers indicate protein sizes in kDa.

As a result, it was confirmed that HIF-1a protein was expressed more effectively by hnRNPA2B1, but was dose-dependently inhibited by MO-2097 treatment (Fig. 9).

Additionally, HeLa CCL2 and HCT116 cells were cultured under hypoxic conditions with DMSO control or 25 and 50 μM MO-2097. 20% _O2- conditioned cells were used as normoxic control. After 24 h, RNA was extracted and cDNA was synthesized. HIF-1α, HK1, MRP1, SLCA5, IL-6, and VEGF mRNA levels were measured by qPCR.

As a result, HK, MRP1, SLC1A5, IL6, and VEGF were confirmed to be among the target genes regulated by HIF-1α inhibition with MO-2097 in HeLa CCL2 and HCT116 cells (Fig. 10).

Since the above genes are related to cancer proliferation, compound 1a of the example (MO-2097) can act as an anticancer compound.

Test Example 4. Confirmation of anticancer effect in 3D spheroid model

The anticancer effect of MO-2097 was verified in a similar microscopic cancer tissue environment under 3D culture conditions. Specifically, HCT116 cells were seeded in low attachment 96-well round bottom plates and treated with DMSO control or 25 and 50 μM MO-2097. The spheroid images in Fig. 11 were acquired using a Zeiss microscope (Scale bar: 100 μm). The measured spheroid area data in Fig. 12 are expressed as mean ± SD, p-value<0.0001 based on the measurement on the last day.

According to Figures 11 and 12, 25 μM MO-2097 treatment reduced the spheroid size and inhibited growth, and 50 μM MO-2097 treatment caused spheroid death, indicating the anticancer activity of MO-2097.

The background and anticancer effects of MO-2097 treated samples were observed. For this purpose, HCT116 spheroids were cultured for 8 days and treated with DMSO or 25 and 50 μM MO-2097. The spheroids were then cultured for 2 more days and fixed in 4% paraformaldehyde. After cryosectioning, the samples were stained with anti-cleaved caspase 3 antibody (green) and Tubulin antibody (red), and the results are shown in Fig. 13. In Fig. 13, the blue signal represents DNA (Scale bar: 100 μm).

As in the cell experiments (Figs. 2, 3, and 4), the possible cause of the anticancer effect of the MO-2097 treated sample was investigated to be cell death. As a result of examining the spheroid sample, it was confirmed that cancer cells were killed by MO-2097 treatment even though the central part of the spheroid was in a hypoxic state (Fig. 13).

Test Example 5. Confirmation of anticancer effect in a mouse in vivo xenograft model

MO-2097 25 mg/kg and 50 mg/kg were injected intraperitoneally into three mice seven times, but no obvious toxicity was observed. Therefore, in vivo experiments were performed at the highest dose (50 mg/kg). When the average tumor size reached 50 mm ³ and was palpable, MO-2097 was injected intraperitoneally into the mice together with vehicle every other day for 15 days.

Figure 14 shows mouse images showing the results of xenograft analysis, and mouse body weight, tumor volume, and relative tumor weight are represented graphically, respectively (Figures 15, 16, and 17) (Vehicle group, n=6; MO-2097 treated group, n=6) (Figure 16 ** p<0.01).

According to Figures 14 and 15, MO-2097 progressively reduced the visceral tumor volume without significant body weight loss, indicating that MO-2097 was well tolerated in mice at the tested doses.

In addition, the tumor size of the vehicle treatment group was 637.04 ± 115.49 mm ³ , and the tumor size of the MO-2097 treatment group was 326.7 ± 51.5 mm ³ , confirming that MO-2097 significantly suppressed the tumor volume by about 49% (Figs. 16 and 17).

Following in vivo xenograft studies, additional tumors were resected and immunohistochemical staining was performed. Figure 18 shows the tumor tissue stained with anti-HIF-1α antibody (green), and Figure 19 shows the tumor tissue stained with anti-cleaved caspase 3 antibody (green). The blue signal in Figures 18 and 19 represents DNA (Scale bar: 200 μm).

According to Figure 18, it can be confirmed that HIF-1α expression was significantly reduced in MO-2097 treated samples compared to vehicle treated samples.

Additionally, MO-2097-treated tumors showed increased expression of cleaved caspase 3, an apoptosis marker, suggesting that MO-2097 is a potent inducer of tumor cell apoptosis (Fig. 19).

Overall, these results confirm that compound 1a (MO-2097) of the present invention plays a role in inhibiting HIF-1α protein expression and tumor cell apoptosis in vivo.

Test Example 6. Confirmation of anticancer activity in organoids derived from colon cancer patients

Anticancer activity varies as clinical trials progress due to patient differences. Therefore, it was determined whether compound 1a (MO-2097) of the present invention could be used to observe patient mutations. An experiment using human colon cancer organoids was designed to determine the effect of MO-2097 in human cancer patients. Organoids were developed from patient samples obtained from Chungbuk National University Hospital, and all samples were prepared according to IRB guidelines (No. P01-202007-31-002), resulting in a total of four colon cancer organoids. Each organoid was treated with various concentrations of MO-2097, and organoid growth rate was measured by MTT after 7 or 10 days (only organoid 1 was cultured for 7 days).

Figure 20 shows the results of microscopic observation of the growth inhibition effect on organoids 1, 2, 3, and 4, respectively, after treatment with MO-2097, and Figure 21 shows a graph showing the relative values of the growth inhibition effect on organoids 1, 2, 3, and 4, respectively, after treatment with MO-2097.

According to Figures 20 and 21, it can be confirmed that MO-2097 at a concentration of 25 μM in organoid 1 inhibited organoid growth by 75%, and MO-2097 at a concentration of 12.5 μM in other organoids inhibited organoid growth by 80%.

Additionally, we observed changes in hnRNPA2B1 and HIF-1α proteins in MO-2097-treated samples, as in previous experiments indicating that MO-2097 inhibits HIF-1α via hnRNPA2B1.

Figure 22 shows the results of blotting with the indicated antibodies after dissolving organoids 1 and 2 in sample buffer. According to Figure 22, among the two samples, hnRNPA2B1 expression was found to be higher in organoid 2, and HIF-1α expression in organoid 2 reacted at a lower concentration.

Figure 23 shows the results of harvesting organoid 2 for qPCR and analyzing each primer set of the target gene. According to Figure 23, it can be confirmed that the target gene expression level of HIF-1α responded to MO-2097 in a dose-dependent manner. Although it was not an artificial hypoxic condition, expression and inhibition of HIF-1α were observed under these experimental conditions.

To determine whether the anticancer effect of MO-2097, which is apoptosis, can be observed, organoid 1 was treated with 25 μM MO-2097 for 24 hours, and then immunostained with the indicated antibodies (Tubulin, anti-cleaved caspase 3, DAPI, and Merge), and observed under a fluorescence microscope (Carl Zeiss), as shown in Fig. 24.

As a result, it can be confirmed that the apoptosis marker staining using organoid samples (Fig. 24), cell experiment results (Fig. 4), and in vivo xenograft experiment results (Figs. 17 and 18) all showed the same results.

Test Example 7. Confirmation of MO-2097’s inhibition of cancer metastasis

In order to confirm the anti-metastatic effect of compound 1a (MO-2097) of the present invention on colon cancer cells DLD-1, various analyses were conducted, and when Sorafenib, which has the same anti-metastatic effect, was used, it was also analyzed and compared.

(1) MTT Assay

The results of MTT analysis of compound 1a (MO-2097) of the present invention and Sorafenib at various concentrations are shown in FIGS. 25 and 26, respectively.

According to FIGS. 25 and 26, the suitable concentration of the 1a compound (MO-2097) of the present invention is 30 μM (cell viability of about 91%), and the concentration of Sorafenib, which shows a similar effect, is 2.5 μM (cell viability of about 92%). Therefore, the comparison will be focused on 30 μM of the 1a compound (MO-2097) of the present invention and 2.5 μM of Sorafenib.

(2) Cell migration assay

The cell migration of the compound 1a (MO-2097) of the present invention and Sorafenib at various concentrations and at various time points was tested, and the results are shown in FIGS. 27 and 28, respectively. The degree to which the space where cells fell off was filled according to each concentration was quantified and shown in a graph (FIG. 29) and Tables 2 and 3 below.

MO-2097 medium Standard deviation Control 1.253531 1.761524 10μM 12.01685 2.041439 20μM 27.86044 2.680239 30μM 44.73384 3.333348

Sorafenib medium Standard deviation Control 5.635802 1.45611 1μM 14.54832 7.429388 2.5μM 36.36038 4.598074 5μM 70.58846 6.548142

According to Figures 27 to 29 and Tables 2 and 3, it can be confirmed that treatment with the compound according to the present invention inhibits metastasis of cancer cells in a concentration-dependent manner.

(3) Migration assay

The results of mobility analysis using crystal violet staining of the 1a compound (MO-2097) of the present invention are shown in FIG. 30, the results of mobility analysis using various concentrations of the 1a compound (MO-2097) of the present invention and Sorafenib are numerically shown in FIG. 31, and the results of mobility analysis using DAPI staining of the 1a compound (MO-2097) of the present invention and Sorafenib are shown in FIG. 32.

According to Figures 30 to 32, it was confirmed that treatment with the compound according to the present invention inhibits metastasis of cancer cells.

(4) Western blot analysis

Figure 33 shows the results of Western blotting of compound 1a (MO-2097) of the present invention and Sorafenib.

According to Figure 33, it was confirmed that treatment with the compound according to the present invention significantly inhibited the expression level of RAF-1.

(5) Conclusion

As a result of the analysis of the above (1) to (3), it was confirmed that the compound 1a (MO-2097) of the present invention has an anti-metastatic effect on colon cancer cells DLD-1, and it was confirmed through the analysis of (4) that the compound (MO-2097) inhibits RAF-1.

Therefore, compound 1a (MO-2097) of the present invention has an inhibitory effect on RAF-1, and also shows an excellent inhibitory effect on cancer cell metastasis, thereby showing excellent effects not only on simple inhibition of cancer cell growth but also on inhibition of cancer metastasis, and thus shows high potential for use as a cancer treatment agent.

Test Example 8. Confirmation of inhibition of cancer metastasis by compounds 1a-1t of the example

Similarly, the following experiments were performed to confirm the anti-metastatic effect of compounds 1a-1t of the present invention.

(1) MTT Assay

First, an MTT assay was performed to confirm the anticancer activity of compounds 1a-1t of the present invention against HeLa CCL2 cells.

Specifically, HeLa CCL2 cells were seeded at 5000 cells/well (100 μl per well) in a 96-well flat bottom plate, and then treated with 20 μl of each of the 1a-1t compounds of the example the next day. Considering that the total volume after compound treatment was 120 μl, the treatment was performed at a concentration 6 times higher than the final concentration. MTT solution was treated 48 hours after treating the example compounds, and the results were measured 2 hours later.

The results of the MTT assay for HeLa CCL2 cells of compounds 1a-1t of the present invention are shown in Figures 34 to 36.

According to FIGS. 34 to 36, among the compounds of the present invention, compounds of Examples 1a-1e, 1g, 1m, 1n, 1p-1t have a cell killing effect on HeLa CCL2 cells, and in particular, compounds of Examples 1a, 1c, 1d, 1m, 1n, 1p-1t have an IC ₅₀ of 30 μM or less, confirming that they have relatively strong anticancer activity.

(2) Cell migration assay

In addition, the cell migration effect of compounds 1a-1t of the present invention on HeLa CCL2 cells was confirmed.

Specifically, HeLa CCL2 cells were seeded in a 24-well plate at 1.5 x 10 ⁴ cells/well (500 μl per well), and the cells in the center of the well were removed the next day using a Scratcher (SPL product_CAT # 201925). After that, the cells were washed twice with DMEM culture medium and suctioned, and then treated with 500 μl of the culture medium containing each of the 1a-1t compounds of the examples. Thereafter, images after 0 hour, 24 hours, and 48 hours were taken under a microscope, and after 48 hours, the culture medium was suctioned and the images were obtained by staining for 20 minutes using Wright stain solution (CAT #45253-250ML-F) (Geimsa staining).

The extent to which the space where cells fell off was filled after each treatment with compounds 1a-1t of the present invention was confirmed through images, and this is shown in Figure 37.

According to FIG. 37, among the exemplary compounds of the present invention, compounds of examples 1a-1e, 1g, 1l, 1m, 1q-1t have an effect of inhibiting the metastasis of cancer cells, and in particular, it can be confirmed that compounds of examples 1a, 1c, 1d, 1m, 1q and 1t exhibit a strong metastasis inhibition effect.

(3) Conclusion

In addition, the anticancer activity and anti-metastatic activity of the compounds 1a-1t of the examples confirmed through the analyses of (1) to (2) above are summarized and shown in Figure 38.

That is, the exemplary compounds according to the present invention exhibit excellent inhibitory effects on cancer cell metastasis, thereby demonstrating excellent effects not only in inhibiting simple cancer cell growth but also in inhibiting cancer metastasis, and thus have high potential for use as cancer treatment agents.

This specification has omitted detailed descriptions of matters that can be sufficiently recognized and inferred by those skilled in the art of the present invention, and various modifications are possible within the scope that does not change the technical idea or essential configuration of the present invention other than the specific examples described in this specification. Accordingly, the present invention can be implemented in a manner other than that specifically described and exemplified in this specification, and this is a matter that can be understood by those skilled in the art of the present invention.

Claims (20)

A compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof: [Chemical Formula 1]

In the above chemical formula 1, R ₁ is a hydrogen atom, -OH, or C ₁ -C ₆ alkyl,

Is

And, R ₆ is independently a hydrogen atom, a halogen atom, -OH, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy, -R ₂ OH,

,

, -NHR ₇ or containing a heteroatom selected from oxygen, nitrogen and sulfur, R ₂ is C ₁ -C ₆ alkyl, R ₃ is a hydrogen atom or C ₁ -C ₆ alkyl, R ₄ and R ₅ are the same or different, and are each independently a hydrogen atom, a substituted or unsubstituted C ₁ -C ₆ alkyl, a substituted or unsubstituted C ₃ -C ₈ heterocycloalkyl, or R ₄ and R ₅ are connected to each other together with one or more carbon atoms or nitrogen atoms to form a substituted or unsubstituted C ₃ -C ₈ heterocycloalkyl, wherein the substitution is substitution with at least one or more C ₁ -C ₃ alkyl, C ₆ -C ₁₂ aryl, -C(=O)O-(C ₁ -C ₆ alkyl), or halobenzyl, R ₇ is a hydrogen atom or C ₁ -C ₆ alkyl, n is an integer from 0 to 5, and when n is 2 or greater, R ₆ are identical or different. In claim 1, the compound represented by the chemical formula 1 is a compound represented by the following chemical formula 2, a compound represented by the chemical formula 1 or a pharmaceutically acceptable salt thereof: [Chemical formula 2]

In the above chemical formula 2, R ₁ is a hydrogen atom, -OH, or C ₁ -C ₆ alkyl, R ₄ and R ₅ are the same or different, and are each independently a hydrogen atom, a substituted or unsubstituted C ₁ -C ₆ alkyl, a substituted or unsubstituted C ₃ -C ₈ heterocycloalkyl, or R ₄ and R ₅ are connected to each other together with one or more carbon atoms or nitrogen atoms to form a substituted or unsubstituted C ₃ -C ₈ heterocycloalkyl, wherein the substitution is substitution with at least one or more C ₁ -C ₃ alkyl, C ₆ -C ₁₂ aryl, -C(=O)O-(C ₁ -C ₆ alkyl), or halobenzyl, R ₆ is independently a hydrogen atom, a halogen atom, -OH, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy, -R ₂ OH,

,

, -NHR ₇ or containing a heteroatom selected from oxygen, nitrogen and sulfur, R ₂ is C ₁ -C ₆ alkyl, R ₇ is a hydrogen atom or C ₁ -C ₆ alkyl, n is an integer from 0 to 4, and when n is 2 or greater, R ₆ are identical or different. In the first paragraph, R ₁ is hydrogen, OH, methyl, ethyl, or isobutyl,

Is

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

, or

A compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof. In the first paragraph, R ₁ is methyl,

Is

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

,

, or

A compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof. In the first paragraph, A compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof, which binds to hnRNPA2B1 and inhibits the translation of HIF-1α, is a compound represented by the chemical formula 1 or a pharmaceutically acceptable salt thereof. In the first paragraph, A compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof, which binds to hnRNPA2B1 and inhibits the activity of RAF-1. In the first paragraph, The compound represented by the above chemical formula 1 is any one selected from the group consisting of the following chemical formulas, a compound represented by the chemical formula 1 or a pharmaceutically acceptable salt thereof: 5-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)benzene-1,3-diol; 4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)phenol; 4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)benzene-1,2-diol; 3-methoxy-5-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)phenol; Methyl 4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)benzoate; 4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)benzoic acid; [4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)phenyl](4-methylpiperazin-1-yl)methanone; 2-Fluoro-4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)phenol; N,N-Dimethyl-4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)benzamide; 2-(4-fluorophenyl)-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran; 2-(3,5-difluorophenyl)-7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran; [4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)phenyl](piperidin-1-yl)methanone; 4-(7-Methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)-N-(1-methylpiperidin-4-yl)benzamide; N-Benzyl-4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)benzamide; tert-Butyl 4-[4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)benzoyl]piperazine-1-carboxylate; [4-(4-fluorobenzyl)piperazin-1-yl][4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)phenyl]methanone; [4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)phenyl](piperazin-1-yl)methanone; 3-[7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)-N-(1-methylpiperidin-4-yl)benzamide; 2-Methyl-4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)-N-(1-methylpiperidin-4-yl)benzamide; and 3-Methyl-4-(7-methyl-5,8-dihydrooxepino[3,2-f]benzofuran-2-yl)-N-(1-methylpiperidin-4-yl)benzamide. A pharmaceutical composition for preventing or treating cancer, comprising a compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof: [Chemical Formula 1]

In the above chemical formula 1, R ₁ is a hydrogen atom, -OH, or C ₁ -C ₆ alkyl,

Is

And, R ₆ is independently a hydrogen atom, a halogen atom, -OH, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy, -R ₂ OH,

,

, -NHR ₇ or containing a heteroatom selected from oxygen, nitrogen and sulfur, R ₂ is C ₁ -C ₆ alkyl, R ₃ is a hydrogen atom or C ₁ -C ₆ alkyl, R ₄ and R ₅ are the same or different, and are each independently a hydrogen atom, a substituted or unsubstituted C ₁ -C ₆ alkyl, a substituted or unsubstituted C ₃ -C ₈ heterocycloalkyl, or R ₄ and R ₅ are connected to each other together with one or more carbon atoms or nitrogen atoms to form a substituted or unsubstituted C ₃ -C ₈ heterocycloalkyl, wherein the substitution is substitution with at least one or more C ₁ -C ₃ alkyl, C ₆ -C ₁₂ aryl, -C(=O)O-(C ₁ -C ₆ alkyl), or halobenzyl, R ₇ is a hydrogen atom or C ₁ -C ₆ alkyl, n is an integer from 0 to 5, and when n is 2 or greater, R ₆ are identical or different. In Article 8, A pharmaceutical composition, wherein the cancer is any one selected from the group consisting of colon cancer, breast cancer, brain cancer, neurological cancer, lung cancer, small cell lung cancer, stomach cancer, liver cancer, blood cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, cutaneous or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, anal cancer, colon cancer, breast cancer, fallopian tube carcinoma, endometrial carcinoma, cervical cancer, vaginal cancer, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, renal cell carcinoma, renal pelvic carcinoma, CNS tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma, and pituitary adenoma. In Article 9, A pharmaceutical composition, wherein the cancer is any one selected from the group consisting of colon cancer, uterine cancer, endometrial carcinoma and cervical cancer A pharmaceutical composition for preventing or treating cancer, comprising a compound represented by the following chemical formula 3 or a pharmaceutically acceptable salt thereof. [Chemical Formula 3]

In Article 11, A pharmaceutical composition, wherein the cancer is any one selected from the group consisting of colon cancer, breast cancer, brain cancer, neurological cancer, lung cancer, small cell lung cancer, stomach cancer, liver cancer, blood cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, cutaneous or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, anal cancer, colon cancer, breast cancer, fallopian tube carcinoma, endometrial carcinoma, cervical cancer, vaginal cancer, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, renal cell carcinoma, renal pelvic carcinoma, CNS tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma, and pituitary adenoma. In Article 12, A pharmaceutical composition, wherein the cancer is any one selected from the group consisting of colon cancer, uterine cancer, endometrial cancer, and cervical cancer. In Article 11, A pharmaceutical composition, wherein the composition inhibits cancer metastasis. A pharmaceutical composition comprising a compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier thereof. A compound according to any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, for use in the prevention, treatment or inhibition of metastasis of cancer. A method for treating cancer, comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof. A method for inhibiting cancer metastasis, comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof. Use of a compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the prevention or treatment of cancer. Use of a compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for preventing or treating cancer metastasis.

Images