Novel quinazoline-benzimidazole derivative or pharmaceutically acceptable salt thereof and use thereof

Abstract

The present invention relates to a quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof and a composition comprising the derivative as an active ingredient for preventing or treating cancer. The quinazoline-benzimidazole derivative of the present invention is activated in cancer cells to inhibit tubulin polymerization and tyrosine kinase activity of the human epidermal growth factor receptor 2 (HER2). Upon administration to a subject, the derivative can arrest the cell cycle of cancer cells, induce apoptosis, and block cell growth signals transmitted via the HER2 receptor. Therefore, the derivative can be used as a dual-target anticancer drug for the prevention or treatment of cancer, particularly HER2-positive breast cancer or triple-negative breast cancer.

Description

Novel quinazoline-benzimidazole derivatives or pharmaceutically acceptable salts thereof and uses thereof

[0001] The present invention relates to a quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof and its use.

[0002] Patients with triple-negative breast cancer (TNBC; ER-, PR-, HER2-) account for 10-15% of all breast cancer patients and lack hormone receptors (ER (estrogen receptor), PR (progesterone receptor)) and the HER2 protein, making them ineligible for hormonal therapy or HER2-targeted therapies. Currently, the standard treatment for triple-negative breast cancer relies entirely on conventional cytotoxic chemotherapy (taxene or anthracycline). Because there are no established targeted therapies, treatment strategies for other subtypes are less diverse than for breast cancer. Furthermore, most patients experience recurrence within 2-3 years after surgery or chemotherapy, and metastases easily develop in other organs such as the lungs, liver, brain, and bones, which reduces the patient's survival rate.

[0003] The five-year overall survival rate for patients diagnosed with stage III (stage-III) cancer is less than 55%, and for patients with advanced metastases, the five-year survival rate is even lower, at less than 30%. Most of these patients ultimately suffer from a very serious disease that ultimately leads to death within several years.

[0004] Microtubules, a key component of the cytoskeleton, are composed of tubulin heteropolymers composed of α and β subunits. Microtubules perform various cellular functions, including intracellular transport, maintaining polarity, intracellular signaling, cell migration, and proliferation. During mitosis, microtubules form a spindle, which aligns chromosomes at the center of the cell and then separates them toward the poles. Failure of the spindle to function properly inhibits cell division, leading to apoptosis. Therefore, microtubules are attracting attention as potential targets for anticancer drugs.

[0005] Drugs that target microtubules are broadly divided into two groups: those that stabilize microtubules and those that destabilize them. First, microtubule stabilizers, such as taxanes, paclitaxel (Taxol), and decetaxel, prevent microtubule depolymerization and enhance microtubule polymerization. Most microtubule-stabilizing agents bind to the taxane-binding site or the overlapping site on β-tubulin. Second, microtubule destabilizing agents, such as colchicine and vinca alkaloids, bind to the colchicine-binding site or the vinca-binding site. These microtubule-targeting agents are effective at lower concentrations than those that affect microtubule polymerization, and both inhibit mitosis. Therefore, there is a pressing need to develop potential tubulin polymerization inhibitors as anticancer agents.

[0006] The HER2 (Human Epidermal Growth Factor Receptor 2) receptor is overexpressed in various cancers, particularly breast and gastric cancer, and is known to be a key factor in promoting cancer cell growth, survival, differentiation, and metastasis. HER2 activation activates intracellular signaling pathways via the receptor's tyrosine kinase domain, primarily involving the RAS / RAF / MEK / ERK and PI3K / AKT / mTOR pathways. These signaling pathways promote tumor formation and progression by inducing rapid cancer cell proliferation and anti-apoptosis (inhibition of cell death).

[0007] Previous research has developed various compounds that inhibit HER2 tyrosine kinase activity, thereby blocking this signaling pathway and effectively suppressing cancer cell growth and metastasis. Representative HER2 tyrosine kinase inhibitors include lapatinib and neratinib. These drugs bind to the ATP-binding site of the HER2 receptor, blocking autophosphorylation of tyrosine residues and thereby inhibiting signal transduction. These drugs are particularly effective treatments for HER2-positive breast cancer patients. However, drug resistance and side effects continue to necessitate the development of novel HER2 tyrosine kinase inhibitors.

[0008] The technical problem to be achieved by the present invention is to provide a quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof.

[0009] In addition, another object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer comprising the quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof as an active ingredient.

[0010] Another object of the present invention is to provide a method for preparing the quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof.

[0011] However, the technical problems to be solved by the present invention are not limited to the problems mentioned above, and other problems not mentioned can be clearly understood by those skilled in the art from the description below.

[0012] To solve the above problem, the present invention provides a quinazoline-benzimidazole derivative represented by the following [chemical formula 1] or a pharmaceutically acceptable salt thereof:

[0013] [Chemical Formula 1]

[0014]

[0015] In the above chemical formula 1,

[0016] R 1 Silver -(CH2) n -A and,

[0017] A is substituted or unsubstituted C3-C 20 Cycloalkyl or C3-C 20 Any one of the heterocycloalkyls,

[0018] When the above cycloalkyl or heterocycloalkyl is substituted, it is substituted with at least one selected from the group consisting of a C1-C6 alkyl group, a hydroxyl group, an amine group, a methoxy group, and a halogen group,

[0019] R 2 is substituted or unsubstituted C3-C 20 Aryl, substituted or unsubstituted C3-C 20 Heteroaryl, substituted or unsubstituted C3-C 20 Cycloalkyl, substituted or unsubstituted C3-C 20 Any one selected from the group consisting of heterocycloalkyl and methyl carbamate,

[0020] When the above aryl, cycloalkyl or heterocycloalkyl is substituted, it is substituted with at least one selected from the group consisting of a C1-C6 alkyl group, a halogen group, a hydroxy group and an alkoxy group, and n is an integer from 0 to 6.

[0021] According to one side,

[0022] In the above [chemical formula 1],

[0023] A is substituted or unsubstituted C3-C 20 is a heterocycloalkyl,

[0024] When the above heterocycloalkyl is substituted, it is substituted with at least one of a methyl group or a hydroxy group,

[0025] R 2 is any one selected from the group consisting of substituted or unsubstituted phenyl, substituted or unsubstituted thiophenyl, substituted or unsubstituted pyridinyl, substituted or unsubstituted cyclopentyl, substituted or unsubstituted cyclohexyl and -NH-(C=O)-OCH3,

[0026] When the above phenyl, thiophenyl, pyridinyl, cyclopentyl and cyclohexyl are substituted, they are substituted with at least one selected from the group consisting of methyl, methoxy, halogen and hydroxy, and n can be 2.

[0027] According to one side,

[0028] In the above [chemical formula 1],

[0029] C3-C of A 20 The heterocycloalkyl may be either morpholine or piperidine.

[0030] According to one side,

[0031] The quinazoline-benzimidazole derivative represented by the above [chemical formula 1] may be any one selected from the group consisting of the following compounds:

[0032] [Chemical Formula 1-1]

[0033]

[0034] [Chemical Formula 1-2]

[0035]

[0036] [Chemical Formula 1-3]

[0037]

[0038] [Chemical Formula 1-4]

[0039]

[0040] [Chemical Formula 1-5]

[0041]

[0042] [Chemical Formula 1-6]

[0043]

[0044] [Chemical Formula 1-7]

[0045]

[0046] [Chemical Formula 1-8]

[0047]

[0048] [Chemical Formula 1-9]

[0049]

[0050] [Chemical Formula 1-10]

[0051]

[0052] [Chemical Formula 1-11]

[0053]

[0054] [Chemical Formula 1-12]

[0055]

[0056] [Chemical Formula 1-13]

[0057]

[0058] [Chemical Formula 1-14]

[0059]

[0060] [Chemical Formula 1-15]

[0061]

[0062] [Chemical Formula 1-16]

[0063]

[0064] [Chemical Formula 1-17]

[0065]

[0066] [Chemical Formula 1-18]

[0067]

[0068] [Chemical Formula 1-19]

[0069]

[0070] [Chemical Formula 1-20]

[0071]

[0072] [Chemical Formula 1-21]

[0073]

[0074] [Chemical Formula 1-22]

[0075]

[0076] [Chemical Formula 1-23]

[0077]

[0078] [Chemical Formula 1-24]

[0079]

[0080] [Chemical Formula 1-25]

[0081]

[0082] [Chemical Formula 1-26]

[0083]

[0084] [Chemical Formula 1-27]

[0085]

[0086] [Chemical Formula 1-28]

[0087]

[0088] [Chemical Formula 1-29]

[0089]

[0090] [Chemical Formula 1-30]

[0091]

[0092] [Chemical Formula 1-31]

[0093]

[0094] According to one side, the quinazoline-benzimidazole derivative may have both an effect of inhibiting tubulin polymerization and an effect of inhibiting tyrosine kinase of human epidermal growth receptor 2 (HER2).

[0095] According to one aspect, the pharmaceutically acceptable salt of the quinazoline-benzimidazole derivative may be at least one selected from the group consisting of hydrochloride, bromate, sulfate, phosphate, nitrate, citrate, acetate, lactate, tartrate, maleate, gluconate, succinate, formate, trifluoroacetate, oxalate, fumarate, glutarate, adipate, methanesulfonate, benzenesulfonate, paratoluenesulfonate, camphorsulfonate, sodium salt, potassium salt, lithium salt, calcium salt, and magnesium salt.

[0096] According to another embodiment of the present invention, a pharmaceutical composition for preventing or treating cancer is provided, comprising the quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof as an active ingredient.

[0097] According to one aspect, the pharmaceutical composition can induce apoptosis by arresting the cancer cell cycle.

[0098] According to one aspect, the pharmaceutical composition can inhibit the activity of any one or more of HER2, p95HER2, HER3, EGFR, AKT and MEK.

[0099] According to one side, the cancer may be at least one selected from the group consisting of skin cancer, breast cancer, uterine cancer, esophageal cancer, stomach cancer, brain tumor, colon cancer, rectal cancer, large intestine cancer, lung cancer, ovarian cancer, cervical cancer, endometrial cancer, vulvar cancer, kidney cancer, blood cancer, pancreatic cancer, prostate cancer, testicular cancer, laryngeal cancer, head and neck cancer, thyroid cancer, liver cancer, bladder cancer, osteosarcoma, lymphoma, blood cancer, thymic cancer, urethral cancer, and bronchial cancer, and preferably may be HER2 positive breast cancer or triple negative breast cancer.

[0100] According to another embodiment of the present invention, a manufacturing method is provided for a quinazoline-benzimidazole derivative represented by the following [chemical formula 1] or a pharmaceutically acceptable salt thereof, characterized in that the quinazoline-benzimidazole derivative is manufactured through the following manufacturing steps:

[0101] 1) R 1 A step of preparing a compound represented by the following [chemical formula 4] by reacting a compound represented by -NH2 with 5-bromoanthranilic acid and triethyl orthoformate;

[0102] 2) A step of preparing a compound represented by [Chemical Formula 2] by reacting a compound represented by [Chemical Formula 4] with either 2-nitro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline or 4-amino-3-nitrophenylboronic acid pinacol ester;

[0103] 3) A step of preparing a compound represented by [Chemical Formula 3] from a compound represented by [Chemical Formula 2]; and

[0104] 4) A compound represented by [chemical formula 3] or R 2A step of producing a compound represented by [chemical formula 1] by reacting with any one of the compounds represented by -CHO;

[0105] [Chemical Formula 1]

[0106]

[0107] [Chemical Formula 2]

[0108]

[0109] [Chemical Formula 3]

[0110]

[0111] [Chemical Formula 4]

[0112]

[0113] In the above chemical formulas 1 to 4,

[0114] R 1 Silver -(CH2) n -A and,

[0115] A is substituted or unsubstituted C3-C 20 Cycloalkyl or C3-C 20 Any one of the heterocycloalkyls,

[0116] When the above cycloalkyl or heterocycloalkyl is substituted, it is substituted with at least one selected from the group consisting of a C1-C6 alkyl group, a hydroxyl group, an amine group, a methoxy group, and a halogen group,

[0117] R 2 is substituted or unsubstituted C3-C 20 Aryl, substituted or unsubstituted C3-C 20 Heteroaryl, substituted or unsubstituted C3-C 20 Cycloalkyl, substituted or unsubstituted C3-C 20 Any one selected from the group consisting of heterocycloalkyl and methyl carbamate,

[0118] When the above aryl, cycloalkyl or heterocycloalkyl is substituted, it is substituted with at least one selected from the group consisting of a C1-C6 alkyl group, a halogen group, a hydroxy group and an alkoxy group, and n is an integer from 0 to 6.

[0119]

[0120] In addition, the present invention provides a composition for preventing or treating cancer, comprising the quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof as an active ingredient.

[0121] In addition, the present invention provides a method for preventing or treating cancer, comprising a step of administering the quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof to a subject.

[0122] Furthermore, the present invention provides the use of the quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the prevention or treatment of cancer.

[0123] In addition, the present invention provides a method for diagnosing cancer, comprising a step of administering the quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof to a subject.

[0124] In addition, the present invention provides the use of the quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof for the manufacture of a drug for diagnosing cancer.

[0125] In one embodiment of the present invention, the pharmaceutical composition may induce apoptosis by arresting the cell cycle of cancer cells, or may block cell growth signals transmitted through the HER2 receptor by inhibiting the tyrosine kinase of human epidermal growth receptor 2 (HER2), and preferably, both mechanisms may be performed.

[0126] In another embodiment of the present invention, the cancer may be at least one selected from the group consisting of skin cancer, breast cancer, uterine cancer, esophageal cancer, stomach cancer, brain tumor, colon cancer, rectal cancer, large intestine cancer, lung cancer, ovarian cancer, cervical cancer, endometrial cancer, vulvar cancer, kidney cancer, blood cancer, pancreatic cancer, prostate cancer, testicular cancer, laryngeal cancer, head and neck cancer, thyroid cancer, liver cancer, bladder cancer, osteosarcoma, lymphoma, leukemia, thymic cancer, urethral cancer, and bronchial cancer, and preferably may be breast cancer, more preferably HER2 positive breast cancer or triple negative breast cancer.

[0127] The present invention relates to a quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof and a composition for preventing or treating cancer comprising the derivative as an active ingredient, wherein the quinazoline-benzimidazole derivative of the present invention is activated in cancer cells to inhibit tubulin polymerization and simultaneously inhibit the tyrosine kinase of human epidermal growth receptor 2 (HER2), thereby blocking the cancer cell cycle, inducing apoptosis, and blocking cell growth signals transmitted through the HER2 receptor, thereby inhibiting cell proliferation and survival and simultaneously obtaining an anticancer effect when administered to a subject. Therefore, the compound of the present invention and the composition comprising the same can be used as a novel dual-target anticancer composition for preventing or treating cancer, preferably for preventing or treating HER2-positive breast cancer or triple-negative breast cancer.

[0128] Figure 1 shows the results of evaluating the change in cell viability through MTS assay after treatment with quinazoline-benzimidazole derivative compounds (1-1 to 1-10) at different concentrations (0, 1, 5 μM) in triple-negative breast cancer cell line MDA-MB-231 and HER2-positive breast cancer cell line JIMT-1.

[0129] Figure 2 shows the results of evaluating the change in cell viability through MTS assay after treatment with quinazoline-benzimidazole derivative compounds (1-11 to 1-20) at different concentrations (0, 1, 5 μM) in triple-negative breast cancer cell line MDA-MB-231 and HER2-positive breast cancer cell line JIMT-1.

[0130] Figure 3 shows the results of evaluating the change in cell viability through MTS assay after treatment with quinazoline-benzimidazole derivative compounds (1-21 to 1-31) at different concentrations (0, 1, 5 μM) in triple-negative breast cancer cell line MDA-MB-231 and HER2-positive breast cancer cell line JIMT-1.

[0131] Figure 4 shows the cell viability and apoptosis morphology of trastuzumab-sensitive and -resistant HER2-positive breast cancer cell lines.

[0132] Figure 5 confirms that tubulin is degraded and apoptosis is induced in SKBR3 and JIMT-1 cell lines.

[0133] Figure 6 confirms G2 / M phase cell cycle arrest and expression of mitotic markers.

[0134] Figure 7 shows the expression and activity of HER2, HER family members, and downstream signaling factors confirmed by Western blot.

[0135] Figure 8 shows the effects of quinazoline-benzimidazole derivative compounds 1-16 and 1-19 on lung cancer, colon cancer, ovarian cancer, prostate cancer, liver cancer, and blood cancer cell lines.

[0136] The present inventors have confirmed that benzimidazole derivatives have anticancer effects by inhibiting tubulin synthesis, thereby arresting the cell cycle and thereby inhibiting cancer cell division, and that quinazoline derivatives can have anticancer effects by inhibiting the tyrosine kinase of human epidermal growth receptor 2 (HER2). In addition, the present inventors have confirmed the possibility of activating both mechanisms of action by combining benzimidazole and quinazoline in a single compound molecule, and thus, they have completed the invention and provide a quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof as a novel dual-target anticancer drug.

[0137] From the above results, the present invention provides a quinazoline-benzimidazole derivative represented by the following [chemical formula 1] or a pharmaceutically acceptable salt thereof:

[0138] [Chemical Formula 1]

[0139]

[0140] In the above chemical formula 1,

[0141] R1 is -(CH2)nA,

[0142] A is either a substituted or unsubstituted C3-C20 cycloalkyl or a C3-C20 heterocycloalkyl,

[0143] When the above cycloalkyl or heterocycloalkyl is substituted, it is substituted with at least one selected from the group consisting of a C1-C6 alkyl group, a hydroxyl group, an amine group, a methoxy group, and a halogen group,

[0144] R2 is any one selected from the group consisting of substituted or unsubstituted C3-C20 aryl, substituted or unsubstituted C3-C20 heteroaryl, substituted or unsubstituted C3-C20 cycloalkyl, substituted or unsubstituted C3-C20 heterocycloalkyl and methyl carbamate,

[0145] When the above aryl, cycloalkyl or heterocycloalkyl is substituted, it is substituted with at least one selected from the group consisting of a C1-C6 alkyl group, a halogen group, a hydroxy group and an alkoxy group, and n is an integer from 0 to 6.

[0146] In the present invention, the term “substitution” refers to a reaction in which an atom or atomic group included in a molecule of a compound is replaced with another atom or atomic group.

[0147] In the present invention, the term “chain-like” refers to a molecule having a chain-like structure, and the chain-like structure is a chemical structure in which carbon atoms are connected in a chain shape, and there are straight chain-like types and branched chain-like types.

[0148] In the present invention, the term “cyclic” refers to a structure in which both ends of the skeleton of an organic compound are connected to form a ring shape.

[0149] In the present invention, the term “straight-chain or branched alkyl group” means a monovalent linear or branched hydrocarbon residue consisting only of carbon and hydrogen atoms, having 1 to 6 carbon atoms.

[0150] In the present invention, the term “cyclic alkyl group” or “cycloalkyl group” means a cyclic saturated hydrocarbon residue.

[0151] Examples of the above alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, 2-butyl, 3-butyl, pentyl, n-hexyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

[0152] In the present invention, the term "heterocycloalkyl group" typically refers to a saturated or unsaturated (but not aromatic) cyclohydrocarbon, which may be optionally unsubstituted, monosubstituted or polysubstituted, and in the structure of which at least one is selected from a heteroatom of N, O or S.

[0153] In the present invention, the term "aryl group" means an unsaturated aromatic ring compound having 3 to 12 carbon atoms, having a single ring (e.g., phenyl) or multiple condensed rings (e.g., naphthyl). Examples of such aryl groups include, but are not limited to, phenyl, naphthyl, and the like.

[0154] In the present invention, the term “heteroaryl group” refers to a single ring or multiple condensed rings in which at least one of the atoms constituting the ring has a heteroatom of N, O, or S. Examples of such heteroaryl groups include, but are not limited to, a pyridyl group, a pyrimidinyl group, a pyrazinyl group, an oxazolyl group, a furyl group, and the like.

[0155] In the present invention, the term “alkoxy group” refers to an alkyl group (-OR) bonded to oxygen. Examples of such alkoxy groups include, but are not limited to, a methoxy group, an ethoxy group, a propoxy group, a butoxy group, and the like.

[0156] In the present invention, the “halogen group” may be fluorine (F), chloride (Cl), bromine (Br), or iodine (I).

[0157] As a preferred embodiment of the present invention, the compound represented by the above chemical formula 1 is preferably at least one selected from the group consisting of the following compounds:

[0158] [Chemical Formula 1-1]

[0159]

[0160] [Chemical Formula 1-2]

[0161]

[0162] [Chemical Formula 1-3]

[0163]

[0164] [Chemical Formula 1-4]

[0165]

[0166] [Chemical Formula 1-5]

[0167]

[0168] [Chemical Formula 1-6]

[0169]

[0170] [Chemical Formula 1-7]

[0171]

[0172] [Chemical Formula 1-8]

[0173]

[0174] [Chemical Formula 1-9]

[0175]

[0176] [Chemical Formula 1-10]

[0177]

[0178] [Chemical Formula 1-11]

[0179]

[0180] [Chemical Formula 1-12]

[0181]

[0182] [Chemical Formula 1-13]

[0183]

[0184] [Chemical Formula 1-14]

[0185]

[0186] [Chemical Formula 1-15]

[0187]

[0188] [Chemical Formula 1-16]

[0189]

[0190] [Chemical Formula 1-17]

[0191]

[0192] [Chemical Formula 1-18]

[0193]

[0194] [Chemical Formula 1-19]

[0195]

[0196] [Chemical Formula 1-20]

[0197]

[0198] [Chemical Formula 1-21]

[0199]

[0200] [Chemical Formula 1-22]

[0201]

[0202] [Chemical Formula 1-23]

[0203]

[0204] [Chemical Formula 1-24]

[0205]

[0206] [Chemical Formula 1-25]

[0207]

[0208] [Chemical Formula 1-26]

[0209]

[0210] [Chemical Formula 1-27]

[0211]

[0212] [Chemical Formula 1-28]

[0213]

[0214] [Chemical Formula 1-29]

[0215]

[0216] [Chemical Formula 1-30]

[0217]

[0218] [Chemical Formula 1-31]

[0219]

[0220] The quinazoline-benzimidazole derivative or pharmaceutically acceptable salt thereof of the present invention is specifically activated in cancer cells to inhibit tubulin polymerization and induce cell death, inhibits the tyrosine kinase of human epidermal growth receptor 2 (HER2) to block cell growth signals transmitted through the HER2 receptor, and inhibits the activity of HER2, p95HER2, HER3, EGFR, AKT, and MEK, and thus can be used as a pharmaceutical composition for preventing or treating cancer containing the derivative as an active ingredient.

[0221] In one embodiment of the present invention, the pharmaceutical composition may induce apoptosis by arresting the cell cycle of cancer cells, or may block cell growth signals transmitted through the HER2 receptor, and preferably, may perform both mechanisms.

[0222] In the present invention, the term “pharmaceutically acceptable salt” means a formulation of a compound that does not cause serious irritation to an organism to which the compound is administered and does not impair the biological activity and physical properties of the compound. The pharmaceutically acceptable salt can be obtained by reacting the compound of the present invention with an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc., a sulfonic acid such as methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, etc., an organic carboxylic acid such as tartaric acid, formic acid, citric acid, acetic acid, trichloroacetic acid, trifluoroacetic acid, capric acid, isobutanoic acid, malonic acid, succinic acid, phthalic acid, gluconic acid, benzoic acid, lactic acid, fumaric acid, maleic acid, salicylic acid, etc. In addition, the compound of the present invention can be obtained by reacting it with a base to form a salt such as an alkali metal salt such as an ammonium salt, a sodium or potassium salt, an alkaline earth metal salt such as a calcium or magnesium salt, a salt of an organic base such as dicyclohexylamine, N-methyl-D-glucamine, tris(hydroxymethyl)methylamine, and a salt of an amino acid such as arginine or lysine.

[0223] In addition, the quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof may include not only a pharmaceutically acceptable salt, but also all salts, hydrates, and solvates that can be prepared by a conventional method.

[0224]

[0225] In addition, the present invention can provide a method for preventing, treating, and / or diagnosing cancer, comprising a step of administering the quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof to a subject.

[0226] In the present invention, the term "prevention" means any act of inhibiting or delaying the occurrence, spread or recurrence of cancer by administering the composition of the present invention, and "treatment" means any act of improving or beneficially changing the symptoms of the disease by administering the composition of the present invention.

[0227] The term "pharmaceutical composition" in the present invention refers to a composition manufactured for the purpose of preventing or treating a disease, and may be formulated and used in various forms according to conventional methods. For example, it may be formulated in oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, and syrups, and may be formulated and used in the form of topical preparations, suppositories, and sterile injectable solutions.

[0228] In the present invention, "including as an active ingredient" means that the ingredient is included in an amount necessary or sufficient to realize a desired biological effect. In actual application, the amount included as an active ingredient can be determined by considering the amount for treating the target disease and not causing other toxicity, and may vary depending on various factors such as the disease or condition being treated, the form of the composition being administered, the size of the subject, or the severity of the disease or condition. A person of ordinary skill in the art to which the present invention pertains can empirically determine the effective amount of an individual composition without undue experimentation.

[0229] In addition, the pharmaceutical composition of the present invention may, depending on each formulation, additionally include one or more pharmaceutically acceptable carriers in addition to the above-described effective ingredients.

[0230] The pharmaceutically acceptable carrier may be saline solution, sterile water, Ringer's solution, buffered saline solution, dextrose solution, maltodextrin solution, glycerol, ethanol, or a mixture of one or more of these components, and may further include other conventional additives such as antioxidants, buffers, and bacteriostatic agents, if necessary. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate the composition into an injectable formulation such as an aqueous solution, suspension, or emulsion, or into a pill, capsule, granule, or tablet. Furthermore, the composition may be preferably formulated according to each disease or component using an appropriate method in the art or a method disclosed in Remington's Pharmaceutical Science (Mack Publishing Company, Easton PA).

[0231]

[0232] The composition of the present invention can be administered orally or parenterally in a pharmaceutically effective amount depending on the intended method, and the term “pharmaceutically effective amount” of the present invention means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment and not causing side effects, and the effective dosage level can be determined based on factors including the patient’s health condition, severity, activity of the drug, sensitivity to the drug, administration method, administration time, administration route and excretion rate, treatment period, drugs used in combination or simultaneously, and other factors well known in the medical field.

[0233] Accordingly, the pharmaceutical composition of the present invention can be administered to a subject to prevent, treat, and / or diagnose cancer, and the cancer may be skin cancer, breast cancer, uterine cancer, esophageal cancer, stomach cancer, brain tumor, colon cancer, rectal cancer, large intestine cancer, lung cancer, ovarian cancer, cervical cancer, endometrial cancer, vulvar cancer, kidney cancer, blood cancer, pancreatic cancer, prostate cancer, testicular cancer, laryngeal cancer, head and neck cancer, thyroid cancer, liver cancer, bladder cancer, osteosarcoma, lymphoma, blood cancer, thymic cancer, urethral cancer, or bronchial cancer, and is not limited thereto, but preferably, it may be a cancer having a characteristic of higher acidity than normal cells and cytotoxicity being inhibited by a tubulin polymerization inhibitor, and non-limiting examples thereof include breast cancer, preferably HER2 positive breast cancer or triple-negative breast cancer.

[0234] In the present invention, the term “subject” is not limited to a mammal such as a livestock or a human that requires prevention, treatment, and / or diagnosis of cancer, but may preferably be a human.

[0235] The pharmaceutical composition of the present invention can be formulated in various forms for administration to a subject, and a representative form of a parenteral administration form is an injectable form, preferably an isotonic aqueous solution or suspension. The injectable form can be prepared using a suitable dispersing or wetting agent and a suspending agent according to a technique known in the art. For example, each component can be dissolved in saline or a buffer to be formulated for injection. In addition, oral administration forms include, for example, ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, and wafers, and these formulations may contain, in addition to the active ingredient, a diluent (e.g., lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, and / or glycine) and a lubricant (e.g., silica, talc, stearic acid and its magnesium or calcium salts, and / or polyethylene glycol). The above tablets may contain a binder such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidine, and optionally may further contain a disintegrating agent such as starch, agar, alginic acid or its sodium salt, an absorbent, a coloring agent, a flavoring agent and / or a sweetening agent. The above formulations may be prepared by conventional mixing, granulating or coating methods.

[0236] In addition, the pharmaceutical composition of the present invention may further include auxiliary agents such as preservatives, wetting agents, emulsifying agents, salts for osmotic pressure control or buffers, and other therapeutically useful substances, and may be formulated according to conventional methods.

[0237] The pharmaceutical composition according to the present invention can be administered via various routes, including oral, transdermal, subcutaneous, intravenous, or intramuscular. The dosage of the active ingredient can be appropriately selected based on various factors, such as the route of administration, the patient's age, sex, weight, and severity of the condition. Furthermore, the composition of the present invention can be administered in combination with known compounds capable of enhancing the desired effect.

[0238] The pharmaceutical composition according to the present invention may be administered to humans or animals orally or parenterally, such as intravenously, subcutaneously, intranasally, or intraperitoneally. Oral administration also includes sublingual administration. Parenteral administration includes injection methods such as subcutaneous injection, intramuscular injection, and intravenous injection, as well as drip methods.

[0239] In the pharmaceutical composition of the present invention, the total effective amount of the quinazoline-benzimidazole derivative or the pharmaceutically acceptable salt thereof according to the present invention can be administered to a patient as a single dose, or can be administered by a fractionated treatment protocol in which multiple doses are administered over a long period of time. The pharmaceutical composition of the present invention may vary the content of the active ingredient depending on the severity of the disease, but can typically be administered repeatedly several times a day at an effective dose of 100 μg to 3,000 mg per administration for adults. However, the concentration of the quinazoline-benzimidazole derivative or the pharmaceutically acceptable salt thereof can be determined by taking into consideration various factors such as the administration route and the number of treatments as well as the patient's age, weight, health status, sex, severity of the disease, diet, and excretion rate.

[0240] In addition, the pharmaceutical composition according to the present invention is not particularly limited in its formulation, administration route, and administration method as long as it exhibits the effects of the present invention, and the pharmaceutical composition of the present invention may additionally include a known anticancer agent in addition to the quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof as an active ingredient, and may be used in combination with other known treatments for the treatment of these diseases.

[0241]

[0242] The terms used in the examples are for illustrative purposes only and should not be construed as limiting. Singular expressions include plural expressions unless the context clearly dictates otherwise. In this specification, terms such as "comprise" or "have" are intended to specify the presence of a feature, number, step, operation, component, part, or combination thereof described in the specification, but should be understood to not preclude the presence or addition of one or more other features, numbers, steps, operations, components, parts, or combinations thereof.

[0243] Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by a person of ordinary skill in the art to which the embodiments pertain. Terms defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning in the context of the relevant technology, and shall not be interpreted in an idealized or overly formal sense unless explicitly defined herein.

[0244]

[0245] Hereinafter, embodiments are described in detail with reference to the attached drawings. However, the embodiments may be modified in various ways, and the scope of the patent application is not limited or restricted by these embodiments. It should be understood that all modifications, equivalents, or alternatives to the embodiments are included within the scope of the patent application.

[0246] In addition, when describing with reference to the attached drawings, identical components will be assigned the same reference numerals regardless of the drawing numbers, and redundant descriptions thereof will be omitted. When describing embodiments, if a detailed description of a related known technology is judged to unnecessarily obscure the gist of the embodiment, the detailed description will be omitted.

[0247]

[0248] [Example]

[0249] Example 1. Preparation of quinazoline-benzimidazole derivatives

[0250] All chemical reagents used were commercially available. 1 H NMR spectra were recorded on a Bruker Avance III 400 MHz, with TMS used as an internal standard.

[0251]

[0252] 1-1. Synthesis of Methyl (6-(3-(2-morpholinoethyl)-4-oxo-3,4-dihydroquinazolin-6-yl)-1H-benzo[d]imidazol-2-yl)carbamate (Representative compound chemical formula 1-1)

[0253]

[0254] Step 1] Synthesis of 6-Bromo-3-(2-morpholinoethyl)quinazolin-4(3H)-one

[0255] 5-Bromoanthranilic acid (2.16 g, 9.3 mmole) was dissolved in 30 ml of anhydrous ethanol under a nitrogen atmosphere. Then, triethyl orthoformate (1.39 g, 13 mmole) and 2-morpholino ethylamine (1.69 g, 13 mmole) were added. Iodine (25 mg, 0.1 mmole) was added as a catalyst. The mixture was heated and refluxed for 6 hours. After confirming the completion of the reaction by TLC, the mixture was cooled to room temperature. The ethanol was removed by distillation under reduced pressure, and the mixture was dissolved in 60 ml of ethyl acetate. The mixture was washed with 1 N aqueous NaOH solution and purified water, dried over anhydrous Na2SO4, and filtered. The filtrate was distilled under reduced pressure to obtain a bright yellow solid (2.21 g).

[0256] Yield 69.7%,

[0257] 1 H NMR (400 MHz. CDCl3): δ 2.51(t, J=3.6 Hz, 4H, 2(N-CH2)), 2.70(t, J=4.76 Hz, 2H, N-CH2), 3.67(t, J=3.6 Hz, 4H, 2(O-CH2)), 4.08(t, J=4.76 Hz, 2H, N-CH2), 7.57~7.84(m, 2H, ArH), 8.05(s, 1H, ArH), 8.43(s, 1H, -NC-H)

[0258]

[0259] Step 2] Synthesis of 6-(4-Amino-3-nitrophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (Formula 2-1)

[0260] 2-Nitro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (1 g, 3.8 mmole), 6-bromo-3-(2-morpholinoethyl)quinazolin-4(3H)-one (1.28 g, 3.8 mmole), and K2CO3 (1.56 g, 11.4 mmole) were dissolved in 25 ml of dioxane under a nitrogen atmosphere. After heating to 80°C, Pd(dppf)Cl2 (0.14 g, 0.19 mmole) was added, and the mixture was heated and refluxed for 6 hours. The completion of the reaction was confirmed by TLC, and distillation under reduced pressure was performed. Then, a yellow solid was obtained through purification by column chromatography. (0.58 g)

[0261] Yield 38.9%,

[0262] 1 H NMR (400 MHz. CDCl3): δ 2.55(t, J=3.6 Hz, 4H, 2(N-CH2)), 2.76(t, J=4.76 Hz, 2H, N-CH2), 3.71(t, J=3.6 Hz, 4H, 2(O-CH2)), 4.14(t, J=4.76 Hz, 2H, N-CH2), 6.24(br. s,, 2H, NH2), 6.95~8.49(m, 7H, Aryl-H)

[0263]

[0264] Step 3] Synthesis of 6-(3,4-Diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (Formula 3-1)

[0265] 6-(4-Amino-3-nitrophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.58 g, 1.4 mmole) was dissolved in 20 ml of methanol, and 0.35 g of 10%-Pd / C was added. The mixture was stirred at room temperature. Hydrazine hydrate (0.75 g, 14 mmole) was added dropwise and the mixture was heated and refluxed for 3 hours. The reaction was confirmed by TLC and cooled to room temperature. After filtration through celite, the filtrate was distilled under reduced pressure. The mixture was then dissolved again in 20 ml of methylene chloride and dried over anhydrous MgSO4. After filtration and distillation under reduced pressure, a yellow solid was obtained. (0.39 g)

[0266] Yield 73.5%,

[0267] 1 H NMR (400 MHz. CDCl3): δ 2.55(t, J=3.6 Hz, 4H, 2(N-CH2)), 2.76(t, J=4.76 Hz, 2H, N-CH2), 3.71(t, J=3.6 Hz, 4H, 2(O-CH2)), 4.14(t, J=4.76 Hz, 2H, N-CH2), 6.24(br. s,, 2H, NH2), 6.81~8.47(m, 7H, Aryl-H)

[0268]

[0269] Step 4] Synthesis of Methyl (6-(3-(2-morpholinoethyl)-4-oxo-3,4-dihydroquinazolin-6-yl)-1H-benzo[d]imidazol-2-yl)carbamate (Formula 1-1)

[0270] 6-(3,4-Diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.2 g, 0.5 mmole) and 1,3-Bis(methoxycarbonyl)-2-methyl-2-thiopseudourea (0.29 g, 14 mmole) were added to 15 ml of 5%-AcOH in ethanol, and then heated under reflux for 3 hours. When a precipitate formed, it was cooled and filtered to obtain a yellow solid (0.17 g).

[0271] Yield 73%

[0272] 1 H NMR (400 MHz. DMSO-d6): δ 2.55(t, J=3.6 Hz, 4H, 2(N-CH2)), 2.76(t, J=4.76 Hz, 2H, N-CH2), 3.71(t, J=3.6 Hz, 4H, 2(O-CH2)), 3.75(s, 3H, -OCH3), 4.14 (t, J=4.76 Hz, 2H, N-CH2), 7.50~8.38 (m, 7H, Aryl-H)

[0273]

[0274] 1-2. Synthesis of 3-(2-Morpholinoethyl)-6-(2-(p-tolyl)-1H-benzo[d]imidazol-6-yl)quinazolin-4(3H)-one (Chemical Formula 1-2)

[0275] 6-(3,4-Diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.15 g, 0.4 mmole) and p-methylbenzaldehyde (50 mg, 0.4 mmole) were dissolved in 10 ml of DMF and stirred at 90°C for 5 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and 10 ml of purified water was washed three times. Then, the mixture was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After purification using column chromatography, a pale yellow solid was obtained. (0.1 g)

[0276] Yield 52.53%

[0277] 1 H NMR (400 MHz. CDCl3): δ 2.38(s, 3H, -CH3), 2.54(t, J=3.6 Hz, 4H, 2(N-CH2)), 2.76(t, J=4.76 Hz, 2H, N-CH2), 3.69(t, J=3.6 Hz, 4H, 2(O-CH2)), 4.14(t, J=4.76 Hz, 2H, N-CH2), 7.26~8.53(m, 7H, Aryl-H)

[0278]

[0279] 1-3. Synthesis of 6-(2-(4-Fluorophenyl)-1H-benzo[d]imidazol-6-yl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (Chemical Formula 1-3)

[0280] 6-(3,4-Diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.15 g, 0.4 mmole) and p-fluorobenzaldehyde (50 mg, 0.4 mmole) were dissolved in 10 ml of DMF and stirred at 90°C for 5 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and 10 ml of purified water was washed three times. Then, it was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After this, it was purified using column chromatography to obtain a pale yellow solid.

[0281] (0.07 g)

[0282] Yield 36.8%,

[0283] 1 H NMR (400 MHz. DMSO-d6): δ 2.33(t, J=3.6 Hz, 4H, 2(N-CH2)), 2.65(t, J=4.76 Hz, 2H, N-CH2), 3.55(t, J=3.6 Hz, 4H, 2(O-CH2)), 4.13(t, J=4.76 Hz, 2H, N-CH2), 7.41~8.40(m, 11H, Aryl-H), 13.08(br. s,, 1H, NH)

[0284]

[0285] 1-4. Synthesis of 6-(2-(3-Fluorophenyl)-1H-benzo[d]imidazol-6-yl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (Chemical Formula 1-4)

[0286] 6-(3,4-Diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.15 g, 0.4 mmole) and m-fluorobenzaldehyde (50 mg, 0.4 mmole) were dissolved in 10 ml of DMF and stirred at 90°C for 5 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and 10 ml of purified water was washed three times. Then, it was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After this, it was purified using column chromatography to obtain a pale yellow solid.

[0287] (0.048 g)

[0288] Yield 22.85%

[0289] 1 H NMR (400 MHz. DMSO-d6): δ 2.33(t, J=3.6 Hz, 4H, 2(N-CH2)), 2.67(t, J=4.76 Hz, 2H, N-CH2), 3.55(t, J=3.6 Hz, 4H, 2(O-CH2)), 4.13(t, J=4.76 Hz, 2H, N-CH2), 7.35~8.41(m, 11H, Aryl-H), 13.19(br. s,, 1H, NH)

[0290]

[0291] 1-5. Synthesis of 6-(2-(4-Chlorophenyl)-1H-benzo[d]imidazol-6-yl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (Formula 1-5)

[0292] 6-(3,4-Diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.15 g, 0.4 mmole) and p-chlorobenzaldehyde (58 mg, 0.4 mmole) were dissolved in 10 ml of DMF and stirred at 90°C for 5 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and 10 ml of purified water was washed three times. Then, the mixture was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After purification using column chromatography, a pale yellow solid was obtained. (0.075 g)

[0293] Yield 37.68%,

[0294] 1 H NMR (400 MHz. DMSO-d6): δ 2.46(t, J=3.6 Hz, 4H, 2(N-CH2)), 2.65(t, J=4.76 Hz, 2H, N-CH2), 3.55(t, J=3.6 Hz, 4H, 2(O-CH2)), 4.14(t, J=4.76 Hz, 2H, N-CH2), 7.62~8.40(m, 11H, Aryl-H), 13.14(br. s, 1H, NH)

[0295]

[0296] 1-6. Synthesis of 6-(2-(2,4-Dihydroxyphenyl)-1H-benzo[d]imidazol-6-yl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (Formula 1-6)

[0297] 6-(3,4-Diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.15 g, 0.4 mmole) and 2,4-dihydroxybenzaldehyde (57 mg, 0.4 mmole) were dissolved in 10 ml of DMF and stirred at 90°C for 5 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and 10 ml of purified water was washed three times. Then, the mixture was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After purification using column chromatography, a pale yellow solid was obtained. (0.032 g)

[0298] Yield 16%

[0299] 1 H NMR (400 MHz. DMSO-d6): δ 2.33(t, J=3.6 Hz, 4H, 2(N-CH2)), 2.67(t, J=4.76 Hz, 2H, N-CH2), 3.54(t, J=3.6 Hz, 4H, 2(O-CH2)), 6.41(s, 1H, -OH), 6.46(d, 1H, -OH), 4.13(t, J=4.76 Hz, 2H, N-CH2), 7.62~8.40(m, 11H, Aryl-H), 13.14(br, s, 1H, NH)

[0300]

[0301] 1-7. Synthesis of 6-(2-(5-Fluoro-2-hydroxyphenyl)-1H-benzo[d]imidazol-6-yl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (kdnc-2316)

[0302] 6-(3,4-Diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.15 g, 0.4 mmole) and 3-fluoro-6-hydroxybenzaldehyde (57 mg, 0.4 mmole) were dissolved in 10 ml of DMF and stirred at 90°C for 5 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and 10 ml of purified water was washed three times. Then, it was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After this, it was purified using column chromatography to obtain a pale yellow solid. (0.091 g)

[0303] Yield 45.73%

[0304] 1 H NMR (400 MHz. DMSO-d6): δ 2.46(t, J=3.6 Hz, 4H, 2(N-CH2)), 2.63(t, J=4.76 Hz, 2H, N-CH2), 3.54(t, J=3.6 Hz, 4H, 2(O-CH2)), 4.13(t, J=4.76 Hz, 2H, N-CH2), 7.06~8.42(m, 11H, Aryl-H), 13.07(br. s,, 1H, NH)

[0305]

[0306] 1-8. Synthesis of 6-(2-Cyclohexyl-1H-benzo[d]imidazol-6-yl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (Formula 1-8)

[0307] 6-(3,4-Diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.15 g, 0.4 mmole) and cyclohexylbenzaldehyde (46 mg, 0.4 mmole) were dissolved in 10 ml of DMF and stirred at 90°C for 5 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and 10 ml of purified water was washed three times. Then, it was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After this, it was purified using column chromatography to obtain a pale yellow solid. (0.030 g)

[0308] Yield 16%,

[0309] 1 H NMR (400 MHz. DMSO-d6): δ 1.23(m, 2H, J=11 Hz, Hexyl-H), 1.33(m, 2H, J=11 Hz, Hexyl-H), 1.58(m, 2H, J=11 Hz, Hexyl-H), 1.71(m, 2H, J=11 Hz, Hexyl-H), 1.82(m, 2H, J=11 Hz, Hexyl-H) 2.46(t, J=3.6 Hz, 4H, 2(N-CH2)), 2.63(t, J=4.76 Hz, 2H, N-CH2), 2.87(m, 1H, J=11 Hz, Hexyl-H), 3.53(t, J=3.6 Hz, 4H, 2(O-CH2)), 4.12(t, J=4.76 Hz, 2H, N-CH2), 7.49~8.35(m, 7H, Aryl-H), 12.26(br, s, 1H, NH)

[0310]

[0311] 1-9. Synthesis of 6-(2-Cyclopentyl-1H-benzo[d]imidazol-6-yl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (Formula 1-9)

[0312] 6-(3,4-Diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.15 g, 0.4 mmole) and cyclopentylbenzaldehyde (40 mg, 0.4 mmole) were dissolved in 10 ml of DMF and stirred at 90°C for 5 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and 10 ml of purified water was washed three times. Then, the mixture was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After purification using column chromatography, a pale yellow solid was obtained. (0.024 g)

[0313] Yield 13.3%

[0314] 1 H NMR (400 MHz. DMSO-d6): δ 1.65(m, 2H, J=4.4 Hz, pentyl-H), 1.78(m, 2H, J=4.4 Hz, pentyl-H), 1.91(m, 2H, J=4.4 Hz, pentyl-H), 2.09(m, 2H, J=4.4) Hz, pentyl-H), 2.46(t, J=3.6 Hz, 4H, 2(N-CH2)), 2.63(t, J=4.76 Hz, 2H, N-CH2), 3.32(m, 1H, J=4.4 Hz, pentyl-H), 3.53(t, J=3.6 Hz, 4H, 2(O-CH2)), 4.12(t, J=4.76 Hz, 2H, N-CH2), 7.51~8.36(m, 7H, Aryl-H), 12.28(br, s, 1H, NH)

[0315]

[0316] 1-10. Synthesis of 6-(2-(4-Methoxyphenyl)-1H-benzo[d]imidazoyl3-(2-morpholinoethyl)quinazolin-4(3H)-one (Formula 1-10)

[0317] 6-(3,4-Diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.13 g, 0.35 mmole) and 2,4-difluorobenzaldehyde (48 mg, 0.35 mmole) were dissolved in 10 ml of DMF and stirred at 90°C for 5 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and 10 ml of purified water was washed three times. Then, the mixture was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After purification using column chromatography, a pale yellow solid was obtained. (0.070 g)

[0318] Yield 41.17%

[0319] 1 H NMR (400 MHz. CDCl3): δ 2.46(t, J=3.6 Hz, 4H, 2(N-CH2)), 2.64(t, J=4.76 Hz, 2H, N-CH2), 3.54(t, J=3.6 Hz, 4H, 2(O-CH2)), 3.85(s, 3H, -OCH3) 4.13(t, J=4.76 Hz, 2H, N-CH2), 7.13~8.39(m, 11H, Aryl-H), 12.90(br. s,, 1H, NH),

[0320]

[0321] 1-11. Synthesis of 6-(2-((3-Chloro-4-fluorophenyl)-1H-benzo[d]imidazoyl3-(2-morpholinoethyl)quinazolin-4(3H)-one (Formula 1-11)

[0322] 6-(3,4-Diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.15 g, 0.4 mmole) and 3-chloro-4-fluorobenzaldehyde (65 mg, 0.4 mmole) were dissolved in 10 ml of DMF and stirred at 90°C for 5 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and 10 ml of purified water was washed three times. Then, it was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After this, it was purified using column chromatography to obtain a pale yellow solid. (0.116 g)

[0323] Yield 56.31%

[0324] 1 H NMR (400 MHz. CDCl3): δ 2.46(t, J=3.6 Hz, 4H, 2(N-CH2)), 2.65(t, J=4.76 Hz, 2H, N-CH2), 3.54(t, J=3.6 Hz, 4H, 2(O-CH2)), 4.13(t, J=4.76 Hz, 2H, N-CH2), 7.63~8.41(m, 10H, Aryl-H), 13.18(br, s, 1H, NH)

[0325]

[0326] 1-12. Synthesis of 6-(2-((2,4-Difluorophenyl)-1H-benzo[d]imidazoyl3-(2-morpholinoethyl)quinazolin-4(3H)-one (Formula 1-12)

[0327] 6-(3,4-Diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.15 g, 0.4 mmole) and 2,4-difluorobenzaldehyde (58 mg, 0.4 mmole) were dissolved in 10 ml of DMF and stirred at 90°C for 5 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and 10 ml of purified water was washed three times. Then, the mixture was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After purification using column chromatography, a pale yellow solid was obtained. (0.057 g)

[0328] Yield 28.5%

[0329] 1 H NMR (400 MHz. CDCl3): δ 2.45(t, J=3.6 Hz, 4H, 2(N-CH2)), 2.65(t, J=4.76 Hz, 2H, N-CH2), 3.55(t, J=3.6 Hz, 4H, 2(O-CH2)), 4.13(t, J=4.76 Hz, 2H, N-CH2), 7.32~8.40(m, 10H, Aryl-H), 12.8(br s, 1H, NH)

[0330]

[0331] 1-13. Synthesis of Methyl (6-(4-oxo-3-(2-(piperidin-1-yl)ethyl)-3,4-dihydroquinazolin-6-yl)-1H-benzo[d]imidazol-2-yl)carbamate (Representative compound chemical formula 1-13)

[0332]

[0333] Step 1] Synthesis of 6-Bromo-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one

[0334] Add 30 ml of anhydrous ethanol to 2-Amino-5-bromobenzoic acid (2.16 g, 9.4 mmole), and then add 1-(2-Aminoethyl)piperidine (1.56 g, 12.2 mmole), triethyl orthoformate (1.8 g, 12.2 mmole), and iodine (23 mg, 0.09 mmole) in that order under nitrogen, and heat and reflux for 6 hours. After confirming the reaction by TLC, cool to room temperature. After distilling all the ethanol under reduced pressure, add 60 ml of ethyl acetate, wash three times with 30 ml of 1N-NaOH, and wash three times with 30 ml of Brine. Dry the organic layer with anhydrous Na2SO4, filter, and distill under reduced pressure to obtain a brown liquid (2.43 g).

[0335] Yield 77.14%,

[0336] 1 H NMR (400 MHz. DMSO-d6): δ 1.32(m, 6H, 3(-CH2)), 2.35(m, 4H, 2(-CH2)), 2.52(t, J=6 Hz, 2H, N-CH2), 4.13(t, J=6 Hz, 2H, N-CH2), 7.57~8.30(m, 4H, Aryl-H)

[0337]

[0338] Step 2] Synthesis of 6-(4-Amino-3-nitrophenyl)-3-(2-(piperidin-1yl)ethyl)quinazolin-4(3H)-one (Formula 2-2)

[0339] 6-Bromo-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (1.0 g, 3.78 mmole), 4-Amino-3-nitrophenylboronic acid pinacol ester (1.27 g, 3.78 mmole), and potassium carbonate (1.56 g, 11.3 mmole) were dissolved in 25 ml of dioxane aqueous solution (dioxane 20 ml + purified water 5 ml), heated to 100 ℃, Pd(dppf)Cl2 (0.14 g, 0.18 mmole) catalyst was added, and heated and refluxed for 6 hours under a nitrogen atmosphere. After confirming the reaction by TLC, solid impurities were removed by celite filtration. After adding an additional 30 ml of ethyl acetate, slurry was performed for 1 hour and filtered to obtain a brown solid.

[0340] (0.68 g)

[0341] Yield 46%

[0342] 1 H NMR (400 MHz. DMSO-d6): δ 1.37(m, 6H, 3(-CH2)), 2.39(m, 4H, 2(-CH2)), 2.56(t, J=6 Hz, 2H, N-CH2), 4.09(t, J=6 Hz, 2H, N-CH2), 7.15~8.31(m, 7H, Aryl-H)

[0343]

[0344] Step 3] Synthesis of 6-(3,4-Diaminophenyl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (Formula 3-2)

[0345] Dissolve 6-(4-Amino-3-nitrophenyl)-3-(2-(piperidin-1yl)ethyl)quinazolin-4(3H)-one (0.62 g, 1.58 mmole) in 20 ml of methanol, add 0.37 g of 10%-Pd / C, and stir. Then, add hydrazine hydrate (0.79 g, 15.8 mmole) and heat under reflux for 4 hours. After confirming the reaction by TLC, filter through celite. Dry the organic layer over anhydrous magnesium sulfate, filter, and distill under reduced pressure to obtain a bright yellow liquid.

[0346] (0.45 g)

[0347] Yield 78.9%

[0348] 1 H NMR (400 MHz. DMSO-d6): δ 1.37(m, 6H, 3(-CH2)), 2.39(m, 4H, 2(-CH2)), 2.57(t, J=6 Hz, 2H, N-CH2), 4.07(t, J=6 Hz, 2H, N-CH2), 4.65(s, 2H, NH2), 4.74(s, 2H, NH2), 6.59~8.94(m, 7H, Aryl-H)

[0349]

[0350] Step 4] Synthesis of Methyl 6-(3,4-dihydro-4-oxo-3-(2-piperidin-1-yl)ethyl)quinazoli-6-yl)-1H-benzo[d]imidazol-2-yl carbamate (Formula 1-13)

[0351] 6-(3,4-Diaminophenyl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.2 g, 0.5 mmole) and 1,3-Bis(methoxycarbonyl)-2-methyl-2-thiopseudourea (0.29 g, 14 mmole) were added to 15 ml of 5%-AcOH in ethanol, and then heated under reflux for 3 hours. When a precipitate formed, it was cooled and filtered to obtain a yellow solid (0.177 g).

[0352] Yield 72%

[0353] 1 H NMR (400 MHz. CDCl3): δ 1.37(br. s,, 2H, cycloalkyl-CH2) 1.45(br. s,, 4H, cycloalkyl-CH2) 2.40(br. s,, 4H, cycloalkyl N-CH2), 2.67(t, J=4.76 Hz, 2H, N-CH2), 3.80(s, 3H, OCH3), 4.10(t, J=4.76 Hz, 2H, N-CH2), 7.49~8.33(m, 7H, Aryl-H), 11.68(br. s,, 1H, NH),

[0354]

[0355] 1-14. Synthesis of 6-(2-(4-Fluorophenyl)-1H-benzo[d]imidazol-6-yl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (Formula 1-14)

[0356] 6-(3,4-Diaminophenyl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.2 g, 0.55 mmole) and 4-fluorobenzaldehyde (68 mg, 0.55 mmole) were dissolved in 10 ml of DMF and stirred at 90°C for 5 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and 10 ml of purified water was washed three times. Then, it was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After this, it was purified using column chromatography to obtain a pale yellow solid. (0.080 g)

[0357] Yield 42.1%

[0358] 1 H NMR (400 MHz. CDCl3): δ 1.37(br. s, 2H, cycloalkyl-CH2) 1.46(br. s,, 4H, cycloalkyl-CH2) 2.41(br, s. 4H, cycloalkyl N-CH2), 2.68(t, J=4.76 Hz, 2H, N-CH2), 4.11(t, J=4.76 Hz, 2H, N-CH2), 7.42~8.40(m, 11H, Aryl-H), 13.10(br. s, 1H, NH),

[0359]

[0360] 1-15. Synthesis of 6-(2-(4-Chlorophenyl)-1H-benzo[d]imidazol-6-yl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (Formula 1-15)

[0361] 6-(3,4-Diaminophenyl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.2 g, 0.55 mmole) and 4-chlorobenzaldehyde (77 mg, 0.55 mmole) were dissolved in 10 ml of DMF and stirred at 90°C for 5 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and 10 ml of purified water was washed three times. Then, it was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After this, it was purified using column chromatography to obtain a pale yellow solid. (0.103 g)

[0362] Yield 38.72%

[0363] 1 H NMR (400 MHz. CDCl3): δ 1.37(br. s,, 2H, cycloalkyl-CH2) 1.45(br. s,, 4H, cycloalkyl-CH2) 2.41(br. s,, 4H, cycloalkyl N-CH2), 2.58(t, J=4.76 Hz, 2H, N-CH2), 4.11(t, J=4.76 Hz, 2H, N-CH2), 7.62~8.40(m, 11H, Aryl-H), 13.23(br. s,, 1H, NH)

[0364]

[0365] 1-16. Synthesis of 6-(2-(3-Chloro-4-chlorophenyl)-1H-benzo[d]imidazol-6-yl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (Formula 1-16)

[0366] 6-(3,4-Diaminophenyl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.2 g, 0.55 mmole) and 3-chloro-4-fluorobenzaldehyde (87 mg, 0.55 mmole) were dissolved in 10 ml of DMF, sodium metabiosulfite (Na2S2O5, 114 mg) was added, and the mixture was stirred at 95°C for 3 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and 10 ml of purified water was washed three times. Then, the mixture was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. Afterwards, the product was purified using silica gel column chromatography by sequentially injecting mixed solutions of methylene chloride and methanol (5:1, v / v), methylene chloride and methanol (3:1, v / v), and ethyl acetate and methanol (5:1, v / v), respectively, and a pale yellow solid was obtained. (0.048 g)

[0367] Yield 17.39%

[0368] 1 H NMR (400 MHz. CDCl3): δ 1.38(br. s,, 2H, cycloalkyl-CH2) 1.46(br. s,, 4H, cycloalkyl-CH2) 2.41(br. s,, 4H, cycloalkyl N-CH2), 2.68(t, J=4.76 Hz, 2H, N-CH2), 4.11(t, J=4.76 Hz, 2H, N-CH2), 7.64~8.40(m, 10H, Aryl-H), 13.19(br. s,, 2H, NH)

[0369]

[0370] 1-17. Synthesis of 6-(2-(2,4-Difliorophenyl)-1H-benzo[d]imidazol-6-yl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (Formula 1-17)

[0371] 6-(3,4-Diaminophenyl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.2 g, 0.55 mmole) and 2,4-fluorobenzaldehyde (78 mg, 0.55 mmole) were dissolved in 10 ml of DMF and stirred at 90°C for 5 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and 10 ml of purified water was washed three times. Then, it was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After this, it was purified using column chromatography to obtain a pale yellow solid. (0.083 g)

[0372] Yield 31.08%

[0373] 1 H NMR (400 MHz. CDCl3): δ 1.36(br. s,, 2H, cycloalkyl-CH2) 1.45(br. s,, 4H, cycloalkyl-CH2) 2.40(br. s,, 4H, cycloalkyl N-CH2), 2.58(t, J=4.76 Hz, 2H, N-CH2), 4.11(t, J=4.76 Hz, 2H, N-CH2), 7.31~8.40(m, 10H, Aryl-H), 12.74(br. s,, 2H, NH)

[0374]

[0375] 1-18. Synthesis of 6-(2-(4-Methoxyphenyl)-1H-benzo[d]imidazol-6-yl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (Formula 1-18)

[0376] 6-(3,4-Diaminophenyl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.2 g, 0.55 mmole) and 4-methoxybenzaldehyde (75 mg, 0.55 mmole) were dissolved in 10 ml of DMF and stirred at 90°C for 5 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and 10 ml of purified water was washed three times. Then, it was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After this, it was purified using column chromatography to obtain a pale yellow solid. (0.084 g)

[0377] Yield 31.93%

[0378] 1 H NMR (400 MHz. CDCl3): δ 1.36 (br. s,, 2H, cycloalkyl-CH2) 1.45 (br. s,, 4H, cycloalkyl-CH2) 2.40(br. s,, 4H, cycloalkyl N-CH2), 2.58(t, J=4.76 Hz, 2H, N-CH2), 3.85(s, 3H, OCH3), 4.12(t, J=4.76 Hz, 2H, N-CH2), 7.12~8.39(m, 11H, Aryl-H), 12.91(br. s,, 1H, NH)

[0379]

[0380] 1-19. Synthesis of 3-(2-Piperidin-1-yl)ethyl)-6-(2-p-toyl-1H-benzo[d]imidazol-6-yl)quinazolin-4(3H)-one (Formula 1-19)

[0381] 6-(3,4-Diaminophenyl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.15 g, 0.41 mmole) and 4-methylbenzaldehyde (49 mg, 0.41 mmole) were dissolved in 10 ml of DMF and stirred at 90°C for 5 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and 10 ml of purified water was washed three times. Then, it was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After this, it was purified using column chromatography to obtain a pale yellow solid. (0.080 g)

[0382] Yield 42.1%

[0383] 1 H NMR (400 MHz. CDCl3): δ 1.38 (br. s,, 2H, cycloalkyl-CH2) 1.45 (br. s,, 4H, cycloalkyl-CH2) 2.40(br. s,, 4H, cycloalkyl N-CH2), 2.41(s, 3H, -CH3), 4.10(t, J=4.76 Hz, 2H, N-CH2), 7.38~8.39(m, 11H, Aryl-H), 12.98(br. s,, 1H, NH)

[0384]

[0385] 1-20. Synthesis of 3-(2-(Piperidin-1-yl)ethyl)-6-(2-(thiophen-3-yl)-1H-benzo[d]imidazol-6-yl)quinazolin-4(3H)-one (Formula 1-20)

[0386] 6-(3,4-Diaminophenyl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.20 g, 0.55 mmole) and 3-Thiophenecarboxaldehyde (62 mg, 0.55 mmole) were dissolved in 10 ml of DMF and stirred at 90°C for 5 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and 10 ml of purified water was washed three times. Then, it was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After this, it was purified using column chromatography to obtain a pale yellow solid. (0.10 g)

[0387] Yield 40.0%

[0388] 1 H NMR (400 MHz. DMSO-d6): δ 1.38 (m, 6H, 3(-CH2)), 2.33(m, 4H, 2(-CH2)), 2.67(t, J=6 Hz, 2H, N-CH2), 4.13(t, J=6 Hz, 2H, N-CH2), 7.57~8.40(m, 10H, Aryl-H), 12.95(br. s,, 1H, NH)

[0389]

[0390] 1-21. Synthesis of 3-(2-(Piperidin-1-yl)ethyl)-6-(2-(pyridin-4-yl)-1H-benzo[d]imidazol-6-yl)quinazolin-4(3H)-one (Formula 1-21)

[0391] 6-(3,4-Diaminophenyl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.2 g, 0.55 mmole) and 4-Pyridinecarboxaldehyde (59 mg, 0.55 mmole) were dissolved in 10 ml of DMF and stirred at 90°C for 5 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and 10 ml of purified water was washed three times. Then, it was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After this, it was purified using column chromatography to obtain a pale yellow solid. (0.03 g)

[0392] Yield 12.14%

[0393] 1 H NMR (400 MHz. DMSO-d6): δ 1.38 (m, 6H, 3(-CH2)), 2.33(m, 4H, 2(-CH2)), 2.67(t, J=6 Hz, 2H, N-CH2), 4.13(t, J=6 Hz, 2H, N-CH2), 7.67~8.80(m, 11H, Aryl-H), 13.43(br. s,, 1H, NH)

[0394]

[0395] 1-22. Synthesis of 6-(2-(5-Chloropyridin-2-yl)-1H-benzo[d]imidazol-6-yl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (Formula 1-22)

[0396] 6-(3,4-Diaminophenyl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.2 g, 0.55 mmole) and 5-chloro-2-pyridinecarboxaldehyde (78 mg, 0.55 mmole) were dissolved in 10 ml of DMF and stirred at 90°C for 5 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and 10 ml of purified water was washed three times. Then, it was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After this, it was purified using column chromatography to obtain a pale yellow solid.

[0397] (0.148 g)

[0398] Yield 55.63%

[0399] 1 H NMR (400MHz. DMSO-d6): δ 1.38(m, 6H, 3(-CH2)), 2.33(m, 4H, 2(-CH2)), 2.60(t, J=6 Hz, 2H, N-CH2), 4.11(t, J=6 Hz, 2H, N-CH2), 7.66~8.83(m, 10H, Aryl-H), 13.35(br. s,, 1H, NH)

[0400]

[0401] 1-23. Synthesis of 3-(2-Morpholinoethyl)-6-(2-(thiophen-3-yl)-1H-benzo[d]imidazol-6-yl)quinazolin-4(3H)-one (Formula 1-23)

[0402] 6-(3,4-Diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.20 g, 0.55 mmole) and 3-Thiophenecarboxaldehyde (61 mg, 0.55 mmole) were dissolved in 10 ml of DMF and stirred at 90°C for 5 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and 10 ml of purified water was washed three times. Then, it was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After this, it was purified using column chromatography to obtain a pale yellow solid. (0.15 g)

[0403] Yield 60.0%

[0404] 1 H NMR (400 MHz. DMSO-d6): δ 2.46(t, J=3.6 Hz, 4H, 2(N-CH2)), 2.65(t, J=4.46 Hz, 2H, N-CH2), 3.54(t, J=4.4 Hz, 4H, 2(O-CH2)), 4.13(t, J=6) Hz, 2H, N-CH2), 7.57~8.39(m, 10H, Aryl-H), 12.95(br. s,, 1H, NH)

[0405]

[0406] 1-24. Synthesis of 3-(2-Morpholinoethyl)-6-(2-(pyridin-4-yl)-1H-benzo[d]imidazol-6-yl)quinazolin-4(3H)-one (Formula 1-24)

[0407] 6-(3,4-Diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.2 g, 0.55 mmole) and 4-Pyridinecarboxaldehyde (58 mg, 0.55 mmole) were dissolved in 10 ml of DMF and stirred at 90°C for 5 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and 10 ml of purified water was washed three times. Then, the mixture was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After purification using column chromatography, a pale yellow solid was obtained. (0.02 g)

[0408] Yield 8.1%

[0409] 1 H NMR (400 MHz. DMSO-d6): δ 2.46(t, J=3.6 Hz, 4H, 2(N-CH2)), 2.65(t, J=4.46 Hz, 2H, N-CH2), 3.54(t, J=4.4 Hz, 4H, 2(O-CH2)), 4.13(t, J=6) Hz, 2H, N-CH2), 7.67~8.8(m, 11H, Aryl-H), 13.43(br. s,, 1H, NH)

[0410]

[0411] 1-25. Synthesis of 6-(2-(5-Chloropyridin-2-yl)-1H-benzo[d]imidazol-6-yl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (Formula 1-25)

[0412] 6-(3,4-Diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.22 g, 0.55 mmole) and 5-chloro-2-pyridinecarboxaldehyde (77 mg, 0.55 mmole) were dissolved in 10 ml of DMF and stirred at 90°C for 5 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and 10 ml of purified water was washed three times. Then, it was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After this, it was purified using column chromatography to obtain a pale yellow solid.

[0413] (0.052 g)

[0414] Yield 19.54%

[0415] 1 H NMR (400 MHz. DMSO-d6): δ 2.46(t, J=3.6 Hz, 4H, 2(N-CH2)), 2.65(t, J=4.46 Hz, 2H, N-CH2), 3.54(t, J=4.4 Hz, 4H, 2(O-CH2)), 4.13(t, J=6) Hz, 2H, N-CH2), 7.63~8.83(m, 10H, Aryl-H), 13.35(br. s,, 1H, NH)

[0416]

[0417] 1-26. Synthesis of 6-(2-(3-chloro-4-fluorophenyl)-1H-benzo[d]imidazol-6-yl)-3-(2-(4-methylpiperidin-1-yl)ethyl)quinazolin-4(3H)-one (Representative compound chemical formula 1-26)

[0418]

[0419] Step 1] Synthesis of 6-Bromo-3-(2-(4-methylpiperidin-1-yl)ethyl)quinazolin-4(3H)-one

[0420] 2-Amino-5-bromobenzoic acid (1.26 g, 5.5 mmole) and 2-(4-methyl-1-piperidinyl)ethanamine (1.0 g, 7.0 mmole) are added to 30 ml of absolute ethanol under a nitrogen atmosphere. Triethyl orthoformate (1.05 g, 7.0 mmole) and iodine (14 mg, 0.01 eq) are added and heated under reflux for 6 hours. After confirming the reaction by TLC, cool slowly. Then, remove all solvent by distillation under reduced pressure. Then, dilute in 60 ml of ethyl acetate and wash with 1N-NaOH solution and Brine. Add anhydrous Na2SO4, dry, and filter under reduced pressure. The filtrate is distilled under reduced pressure to remove all of it, obtaining a brown liquid.

[0421] (1.69 g)

[0422] Yield 88.48%

[0423] 1 H NMR (400 MHz. DMSO-d6): δ 0.85 (d, J=6.4 Hz, 3H, -CH3), 1.01 (m, 2H, -CH2), 1.27 (m, 1H, -CH), 1.54 (m, 2H, -CH2), 1.93 (m, 2H, -CH2), 2.55(t, J=6 Hz, 2H, -CH2), 2.83(m, 2H, -CH2), 4.06(t, J=6 Hz, 2H, -CH2), 7.61~8.32(m, 4H, Aryl-H)

[0424]

[0425] Step 2] Synthesis of 6-(4-Amino-3-nitrophenyl)-3-(2-(4-methylpiperidin-1-yl)ethyl)quinazolin-4(3H)-one (Formula 2-3)

[0426] Add 2-Nitro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (1.27 g, 4.8 mmole), 6-bromo-3-(2-(4-methylpiperidin-1-yl)ethyl)quinazolin-4(3H)-one (1.68 g, 4.8 mmole) and K2CO3 (1.99 g, 14.4 mmole), add 25 ml of wet Dioxane and stir under a nitrogen atmosphere.

[0427] After that, [1,1'-Bis(diphenylphosphino)ferrocene]palladium(II) dichloride (0.175 g, 0.05 eq) was added and heated under reflux for 5 hours. After that, the reaction was confirmed by TLC and slowly cooled. After cooling, the filtrate was filtered through celite and distilled under reduced pressure. Then, ethyl acetate was added, stirred, and filtered to obtain a brown solid.

[0428] (0.71 g)

[0429] Yield 36.4%

[0430] 1 H NMR (400 MHz. DMSO-d6): δ 0.85 (d, J=6.4 Hz, 3H, -CH3), 1.07 (m, 2H, -CH2), 1.30 (m, 1H, -CH), 1.56 (m, 2H, -CH2), 1.96 (m, 2H, -CH2), 2.58(t, J=6 Hz, 2H, -CH2), 2.87(m, 2H, -CH2), 4.09(t, J=6 Hz, 2H, -CH2), 7.16~8.31(m, 7H, Aryl-H)

[0431]

[0432] Step 3] Synthesis of 6-(3,4-Diaminophenyl)-3-(2-(4-methylpiperidin-1-yl)ethyl)quinazolin-4(3H)-one (Formula 3-3)

[0433] Dissolve 6-(4-Amino-3-nitrophenyl)-3-(2-(4-methylpiperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.71 g, 1.74 mmole) in 28 ml of methanol, add 0.35 g of 10%-Pd / C, and stir. Then, add hydrazine hydrate (0.84 g, 17.4 mmole) and heat to reflux for 7 hours. After confirming the reaction by TLC, cool to room temperature. Filter through celite and distill under reduced pressure. Dissolve in 20 ml of methylene chloride and dry using anhydrous magnesium sulfate. Filtration and distillation under reduced pressure yield a brown liquid.

[0434] (0.27 g)

[0435] Yield 41.5%

[0436] 1 H NMR (400 MHz. DMSO-d6): δ 0.85(d, 3H, -CH3), 1.05(m, 2H, -CH2), 1.29(m, 1H, CH-CH3), 1.55(m, 2H, -CH2), 1.95(m, 2H, -CH2), 2.58(t, J=6) Hz, 2H, -CH2), 2.86(m, 2H, -CH2), 4.07(t, J=6 Hz, 2H, -CH2), 4.64(s, 2H, -NH2), 4.73(s, 2H, -NH2), 6.60~8.21(m, 7H, Aryl-H)

[0437]

[0438] Step 4] Synthesis of 6-(2-(3-Chloro-4-fluorophenyl)-1H-benzo[d]imidazol-6-yl)-3-(2-(4-methylpiperidin-1-yl)ethyl)quinazolin-4(3H)-one (Formula 1-26)

[0439] 6-(3,4-Diaminophenyl)-3-(2-(4-methylpiperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.135 g, 0.35 mmole) and 3-chloro-4-fluorobenzaldehyde (56 mg, 0.35 mmole) were dissolved in 10 ml of DMF, sodium metabiosulfite (Na2S2O5, 85 mg) was added, and the mixture was stirred at 95 degrees for 3 hours. After confirming the reaction by TLC, it was cooled, 30 ml of ethyl acetate was added, and then 10 ml of purified water was washed three times. Then, it was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After this, it was purified using column chromatography to obtain a pale yellow solid.

[0440] (55 mg)

[0441] Yield 30.5%

[0442] 1 H NMR (400 MHz. DMSO-d6): δ 0.87(d, 3H, -CH3), 1.05(m, 2H, CH2), 1.40(m, 1H, -CH-CH3), 1.99(m, 2H, -CH2), 2.33(m, 4H, 2(-CH2)), 2.67(t, J=6 Hz, 2H, N-CH2), 2.86(m, 2H, CH2), 4.11(t, J=6 Hz, 2H, N-CH2), 7.64~8.41(m, 10H, Aryl-H), 13.19(br. s,, 1H, NH)

[0443]

[0444] 1-27. Synthesis of 6-(2-(4-Chlorophenyl)-1H-benzo[d]imidazol-6-yl)-3-(2-(4-methylpiperidin-1-yl)ethyl)quinazolin-4(3H)-one (Formula 1-27)

[0445] 6-(3,4-Diaminophenyl)-3-(2-(4-methylpiperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.135 g, 0.35 mmole) and 4-chlorobenzaldehyde (50 mg, 0.35 mmole) were dissolved in 10 ml of DMF, sodium metabiosulfite (Na2S2O5, 85 mg) was added, and the mixture was stirred at 95°C for 3 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and 10 ml of purified water was washed three times. Then, it was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After this, it was purified using column chromatography to obtain a pale yellow solid. (64 mg)

[0446] Yield 35.9%

[0447] 1 H NMR (400 MHz. DMSO-d6): δ 0.87(d, 3H, -CH3), 1.10(m, 2H, CH2), 1.35(m, 1H, -CH-CH3), 1.56(m, 2H, CH2), 1.99(m, 2H, -CH2), 2.33(m, 4H, 2(-CH2)), 2.67(t, J=6 Hz, 2H, N-CH2), 2.86(m, 2H, CH2), 4.10(t, J=6 Hz, 2H, N-CH2), 7.64~8.41(m, 11H, Aryl-H), 13.16(br. s,, 1H, NH)

[0448]

[0449] 1-28. Synthesis of 6-(2-(3-chloro-4-fluorophenyl)-1H-benzo[d]imidazol-6-yl)-3-(2-(4-hydroxypiperidin-1-yl)ethyl)quinazolin-4(3H)-one (Representative compound chemical formula 1-28)

[0450]

[0451] Step 1] Synthesis of 6-Bromo-3-(2-(4-hydroxypiperidin-1-yl)ethyl)quinazolin-4(3H)-one

[0452] 2-Amino-5-bromobenzoic acid (1.26 g, 5.5 mmole) and 1-(2-aminoethyl)-4-piperidinol (1.0 g, 7.0 mmole) are added to 30 ml of absolute ethanol under a nitrogen atmosphere. Triethyl orthoformate (1.05 g, 7.0 mmole) and iodine (14 mg, 0.01 eq) are added and heated under reflux for 6 hours. After confirming the reaction by TLC, cool slowly. Then, remove all solvent by distillation under reduced pressure. Then, dilute in 60 ml of ethyl acetate and wash with 1N-NaOH solution and brine. Add anhydrous Na2SO4, dry, and filter under reduced pressure. The filtrate is distilled under reduced pressure to remove all of it, obtaining a pale yellow solid.

[0453] (1.33 g)

[0454] Yield 69.2%

[0455] 1 H NMR (400 MHz. DMSO-d6): δ 1.28 (m, 2H, -CH2), 1.69 (m, 2H, -CH2), 2.08 (m, 2H, -CH2), 2.55 (t, J=6 Hz, 2H, -CH2), 2.71 (m, 2H, -CH2), 3.42(m, 1H, CH-OH), 4.06(t, J=6 Hz, 2H, -CH2), 4.52(d, J=4.4 Hz, 1H, -OH), 7.63~8.33(m, 4H, Aryl-H)

[0456]

[0457] Step 2] Synthesis of 6-(4-Amino-3-nitrophenyl)-3-(2-(4-hydroxypiperidin-1-yl)ethyl)quinazolin-4(3H)-one (Formula 2-4)

[0458] Add 6-Bromo-3-(2-(4-hydroxypiperidin-1-yl)ethyl)quinazolin-4(3H)-one (1.33 g, 3.78 mmole) and 2-Nitro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (1.0 g, 3.78 mmole), K2CO3 (1.56 g, 11.3 mmole), Pd(dppf)Cl2 (0.13 g, 0.18 mmole) wet Dioxane 25 ml and stir under a nitrogen atmosphere.

[0459] After that, [1,1'-Bis(diphenylphosphino)ferrocene]palladium(II) dichloride (0.13 g, 0.05 eq) was added and heated under reflux for 5 hours. After that, the reaction was confirmed by TLC and cooled slowly. After cooling, the filtrate was filtered through celite and distilled under reduced pressure. Then, ethyl acetate was added, stirred, and filtered to obtain a reddish brown solid.

[0460] (1.2 g)

[0461] Yield 77.4%

[0462] 1 H NMR (400 MHz. DMSO-d6): δ 1.31(m, 2H, (-CH2)), 1.65(m, 2H, (-CH2)), 2.09(m, 2H, (-CH2)), 2.58(t, J=6 Hz, 2H, N-CH2), 2.74(m, 2H, (-CH2)), 4.08(t, J=6 Hz, 2H, N-CH2), 4.53(br. s,, 1H, -OH), 7.15~8.31(m, 7H, Aryl-H)

[0463]

[0464] Step 3] Synthesis of 6-(3,4-Diaminophenyl)-3-(2-(4-hydroxypiperidin-1-yl)ethyl)quinazolin-4(3H)-one (Formula 3-4)

[0465] Dissolve 6-(4-Amino-3-nitrophenyl)-3-(2-(4-hydroxypiperidin-1-yl)ethyl)quinazolin-4(3H)-one (1.55 g, 3.7 mmole) in 80 ml of methanol, add 0.77 g of 10%-Pd / C, and stir. Then, add hydrazine hydrate (1.89 g, 37.8 mmole) and heat to reflux for 7 hours. After confirming the reaction by TLC, cool to room temperature. Filter through celite and distill under reduced pressure. Dissolve in 20 ml of methylene chloride and dry using anhydrous magnesium sulfate. Filtration and distillation under reduced pressure yield a brown solid.

[0466] (1.16 g)

[0467] Yield 80.5%

[0468] 1 H NMR (400 MHz. DMSO-d6): δ 1.32(m, 2H, (-CH2)), 1.69(m, 2H, (-CH2)), 2.13(m, 2H, (-CH2)), 2.61(t, J=6 Hz, 2H, N-CH2), 2.78(m, 2H, (-CH2)), 4.08(t, J=6 Hz, 2H, N-CH2), 4.59(m, 4H, 2(NH2)), 4.72(br. s,, 1H, -OH), 6.60~8.23(m, 7H, Aryl-H)

[0469]

[0470] Step 4] Synthesis of 6-(2-(3-Chloro-4-fluorophenyl)-1H-benzo[d]imidazol-6-yl)-3-(2-(4-hydroxypiperidin-1-yl)ethyl)quinazolin-4(3H)-one (Formula 1-28)

[0471] 6-(3,4-Diaminophenyl)-3-(2-(4-hydroxypiperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.15 g, 0.39 mmole) and 3-chloro-4-fluorobenzaldehyde (63 mg, 0.39 mmole) were dissolved in 10 ml of DMF, sodium metabiosulfite (Na2S2O5, 100 mg) was added, and the mixture was stirred at 95 degrees for 3 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and then 10 ml of purified water was washed three times. Then, it was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After this, the product was purified using column chromatography to obtain a yellow solid.

[0472] (33 mg)

[0473] Yield 16.1%

[0474] 1 H NMR (400 MHz. DMSO-d6): δ 1.31(m, 2H, (-CH2)), 1.69(m, 2H, (-CH2)), 2.11(m, 2H, (-CH2)), 2.61(t, J=6 Hz, 2H, N-CH2), 2.77(m, 2H, (-CH2)), 3.41(m, 1H, -CH-OH), 4.10(t, J=6 Hz, 2H, N-CH2), 4.55(d, 1H, -OH), 7.64~8.42(m, 10H, Aryl-H), 13.20(br. s,, 1H, NH)

[0475]

[0476] 1-29. Synthesis of 6-(2-(4-Chlorophenyl)-1H-benzo[d]imidazol-6-yl)-3-(2-(4-hydroxypiperidin-1-yl)ethyl)quinazolin-4(3H)-one (Formula 1-29)

[0477] 6-(3,4-Diaminophenyl)-3-(2-(4-hydroxypiperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.15 g, 0.39 mmole) and 4-chlorobenzaldehyde (56 mg, 0.39 mmole) were dissolved in 10 ml of DMF, sodium metabiosulfite (Na2S2O5, 100 mg) was added, and the mixture was stirred at 95°C for 3 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and 10 ml of purified water was washed three times. Then, it was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After this, it was purified using column chromatography to obtain a pale yellow solid. (0.48 mg)

[0478] Yield 24.3%

[0479] 1 H NMR (400 MHz. DMSO-d6): δ 1.31(m, 2H, (-CH2)), 1.69(m, 2H, (-CH2)), 2.10(m, 2H, (-CH2)), 2.60(t, J=6 Hz, 2H, N-CH2), 2.77(m, 2H, (-CH2)), 4.09(t, J=6 Hz, 2H, N-CH2), 4.57(s, 1H, -OH), 7.61~8.39(m, 11H, Aryl-H), 13.24(br. s,, 1H, NH)

[0480]

[0481] 1-30. Synthesis of 3-(2-(4-Hydroxypiperidin-1-yl)ethyl)-6-(2-p-tolyl-1H-benzo[d]imidazol-6-yl)quinazolin-4(3H)-one (Formula 1-30)

[0482] 6-(3,4-Diaminophenyl)-3-(2-(4-hydroxypiperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.15 g, 0.39 mmole) and 4-methylbenzaldehyde (47 mg, 0.39 mmole) were dissolved in 10 ml of DMF, sodium metabiosulfite (Na2S2O5, 100 mg) was added, and the mixture was stirred at 95°C for 3 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and 10 ml of purified water was washed three times. Then, it was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After this, the product was purified using column chromatography to obtain a pale yellow solid. (20 mg)

[0483] Yield 10.5%

[0484] 1 H NMR (400 MHz. DMSO-d6): δ 1.31(m, 2H, (-CH2)), 1.69(m, 2H, (-CH2)), 2.10(m, 2H, (-CH2)), 2.40(s, 3H, -CH3), 2.59(t, J=6 Hz, 2H, N-CH2), 2.76(m, 2H, (-CH2)), 4.10(t, J=6 Hz, 2H, N-CH2), 4.55(s, 1H, -OH), 7.37~8.39(m, 11H, Aryl-H), 13.02(br. s,, 1H, NH)

[0485]

[0486] 1-31. Synthesis of 3-(2-(4-Hydroxypiperidin-1-yl)ethyl)-6-(2-(4-methoxyphenyl)-1H-benzo[d]imidazol-6-yl)quinazolin-4(3H)-one (Formula 1-31)

[0487] 6-(3,4-Diaminophenyl)-3-(2-(4-hydroxypiperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.15 g, 0.39 mmole) and 4-methoxybenzaldehyde (54 mg, 0.39 mmole) were dissolved in 10 ml of DMF and stirred at 90°C for 5 hours. After confirming the reaction by TLC, the mixture was cooled, 30 ml of ethyl acetate was added, and 10 ml of purified water was washed three times. Then, it was dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. After this, it was purified using column chromatography to obtain a pale yellow solid.

[0488] (21 mg)

[0489] Yield 10.7%

[0490] 1 H NMR (400 MHz. DMSO-d6): δ 1.34(m, 2H, (-CH2)), 1.69(m, 2H, (-CH2)), 2.10(m, 2H, (-CH2)), 2.59(t, J=6 Hz, 2H, N-CH2), 2.76(m, 2H, (-CH2)), 3.85(s, 3H, -OCH3), 4.10(t, J=6 Hz, 2H, N-CH2), 4.56(s, 1H, -OH), 7.12~8.39(m, 11H, Aryl-H), 12.96(br. s,, 1H, NH)

[0491]

[0492] Example 2. Measurement of anticancer activity and confirmation of mechanism of action of quinazoline-benzimidazole derivatives.

[0493] 2.1 Cell viability screening of quinazoline-benzimidazole derivatives

[0494] MDA-MB-231, a human triple-negative breast cancer (TNBC) cell line (Cell seeding numbers: 0.8(M231) x 10 4cells / wells (confluency 25%) and HER2 positive breast cancer (HER2+ BC) cell line JIMT-1 (Cell seeding numbers: 0.8(JIMT) x 10 4 cells / wells (confluency 25%)) was used in the experiment.

[0495] The above cell lines were cultured in Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum (FBS), streptomycin-penicillin (100 U / mL), and Fungizone (0.625 ㎍ / mL) at 5% CO2 and 37°C.

[0496] The human breast cancer cell lines MDA-MB-231 and JIMT-1 were treated with quinazoline-benzimidazole derivatives of Chemical Formulas 1-1 to 1-31 at various concentrations of 0, 1, and 5 μM, respectively, for 72 hours, and then cell viability was measured using the MTS assay technique. In the MTS assay, cells were allowed to attach to a 96-well plate for 24 hours, treated with the quinazoline-benzimidazole derivatives for 72 hours, and then color was developed with MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) for 4 hours, and then the absorbance was measured at 490 nm using a Spectramax Plus384 microplate analyzer.

[0497] The measurement results are shown in Figures 1 to 3. As a result, it was confirmed that most of the quinazoline-benzimidazole derivatives of the chemical formulas 1-1 to 1-31 inhibited cell viability in a concentration-dependent manner in the human breast cancer cell lines MDA-MB-231 and JIMT-1.

[0498]

[0499] 2.2 Confirmation of the apoptotic effect of quinazoline-benzimidazole derivatives

[0500] The compounds of chemical formulae 1-15, 1-16, 1-19, and 1-22, which showed excellent activity in Example 2-1, were tested as follows. Trastuzumab-sensitive HER2-positive breast cancer cell lines SKBR3 and trastuzumab-resistant JIMT-1 were treated with the compounds at various concentrations (0, 0.1, 0.5, 1, 2, 5 μM) for 72 hours, and the concentration-dependent inhibition of cell viability (MTS assay) and the IC50 value according to the inhibition of cell viability were confirmed, which are shown in Fig. 4A. In addition, after treating SKBR3 and JIMT-1 cell lines with compounds 1-15, 1-16, 1-19, and 1-22 at a concentration of 5 μM for 48 hours, morphological changes in cells were observed using an optical microscope, and this is shown in Figure 4B. It was confirmed that breast cancer cell lines exhibited typical apoptotic morphology by treatment with quinazoline-benzimidazole derivatives.

[0501] Next, SKBR3 and JIMT-1 cell lines were treated with compounds of chemical formulas 1-16 and 1-19 at a concentration of 5 μM for 48 hours, and the degree of cell death was measured by Annexin V / PI staining using flow cytometry, which is shown in Fig. 5A. It was confirmed that the quinazoline-benzimidazole derivatives effectively induced early and late apoptosis in both cell lines. After the same treatment as above, the expression of apoptosis-related proteins and the degradation of tubulin protein were confirmed by Western blotting, which is shown in Fig. 5B. This confirmed that the compounds of the present invention can exhibit apoptotic effects by inhibiting tubulin polymerization.

[0502]

[0503] 2.3 Confirmation of the mechanism of action of quinazoline-benzimidazole derivatives

[0504] The quinazoline-benzimidazole derivative of the present invention is characterized in that, as described above, the quinazoline portion of the compound is designed to provide strong binding to the tyrosine kinase of human epidermal growth receptor 2 (HER2), and at the same time, the benzimidazole portion binds between tubulin polymers to have the ability to inhibit polymerization reaction.

[0505] To confirm this, the following experiment was conducted. After treating trastuzumab-resistant HER2-positive breast cancer cells JIMT-1 with compounds of chemical formulas 1-16 and 1-19 at a concentration of 5 μM for 48 hours, the cells were stained with propidium iodide (PI) and analyzed for cell cycle according to DNA content using flow cytometry, which is shown in Fig. 6A. G2 / M phase cell cycle arrest by treatment with quinazoline-benzimidazole derivatives was confirmed in JIMT-1.

[0506] In addition, after treating JIMT-1 cell lines with compounds 1-16 and 1-19 at a concentration of 5 μM for 24 hours, the expression of phospho-Histone H3, a marker of mitosis, was confirmed through immunocytochemistry, and this is shown in Figure 6B. It was confirmed that the expression of abnormal phospho-Histone H3 protein increased by treatment with quinazoline-benzimidazole derivatives, and thus it was confirmed that the quinazoline-benzimidazole derivatives of the present invention can induce cell cycle arrest through the ability to inhibit tubulin polymerization.

[0507] And, after treatment of JIMT-1 cell line with compounds of chemical formula 1-16 and 1-19 at a concentration of 5 μM for 24, 48 and 72 hours, protein expression and phosphorylation of HER2 and HER family members (HER3, EGFR), AKT and MEK of downstream signaling pathways were confirmed through Western blot technique and are shown in Fig. 7. It was confirmed that the expression and phosphorylation of major proteins in the time-dependent HER2 signaling pathway were reduced by treatment with the quinazoline-benzimidazole derivatives of the present invention, and in particular, the activity of HER2, p95HER2, HER3, EGFR, AKT and MEK, which are the main causes of trastuzumab resistance, were effectively inhibited. Thus, it was confirmed that the quinazoline-benzimidazole derivatives of the present invention have an inhibitory effect on HER2 tyrosine kinase as described above, as per the design goal.

[0508]

[0509] Example 3. Confirmation of anticancer activity of quinazoline-benzimidazole derivatives by cancer type.

[0510] After treating lung cancer cell line H1299, colon cancer cell line HCT116, ovarian cancer cell line SKOV3, prostate cancer cell line Du145, liver cancer cell line HepG2, and blood cancer cell line HL-60 with quinazoline-benzimidazole derivative compounds 1-16 and 1-19 at various concentrations (0, 0.1, 0.5, 1, 5, 10 μM) for 72 hours, concentration-dependent inhibition of cell viability (MTS assay) and IC50 values ​​according to inhibition of cell viability were confirmed, which are shown in Fig. 8. As shown in the figure, it was confirmed that cell viability was effectively inhibited in all other cancer cell lines according to treatment with the quinazoline-benzimidazole derivative of the present invention.

[0511]

[0512] Although the embodiments described above have been described with limited drawings, those skilled in the art will appreciate that various technical modifications and variations can be applied based on the above. For example, appropriate results can still be achieved even if the described techniques are performed in a different order than described, and / or components of the described systems, structures, devices, circuits, etc. are combined or combined in a different manner than described, or are replaced or substituted with other components or equivalents.

[0513] Therefore, other implementations, other embodiments, and equivalents to the claims also fall within the scope of the claims described below.

Claims

1. A quinazoline-benzimidazole derivative represented by the following [chemical formula 1] or a pharmaceutically acceptable salt thereof: [Chemical Formula 1] In the above chemical formula 1, R 1 Silver -(CH2) n -A and, A is substituted or unsubstituted C3-C 20 Cycloalkyl or C3-C 20 Any one of the heterocycloalkyls, When the above cycloalkyl or heterocycloalkyl is substituted, it is substituted with at least one selected from the group consisting of a C1-C6 alkyl group, a hydroxyl group, an amine group, a methoxy group, and a halogen group, R 2 is substituted or unsubstituted C3-C 20 Aryl, substituted or unsubstituted C3-C 20 Heteroaryl, substituted or unsubstituted C3-C 20 Cycloalkyl, substituted or unsubstituted C3-C 20 Any one selected from the group consisting of heterocycloalkyl and methyl carbamate, When the above aryl, cycloalkyl or heterocycloalkyl is substituted, it is substituted with at least one selected from the group consisting of a C1-C6 alkyl group, a halogen group, a hydroxyl group and an alkoxy group, n is an integer from 0 to 6.

2. In paragraph 1, In the above [chemical formula 1], A is substituted or unsubstituted C3-C 20 is a heterocycloalkyl, When the above heterocycloalkyl is substituted, it is substituted with at least one of a methyl group or a hydroxy group, R 2 is any one selected from the group consisting of substituted or unsubstituted phenyl, substituted or unsubstituted thiophenyl, substituted or unsubstituted pyridinyl, substituted or unsubstituted cyclopentyl, substituted or unsubstituted cyclohexyl and -NH-(C=O)-OCH3, In the case where the above phenyl, thiophenyl, pyridinyl, cyclopentyl and cyclohexyl are substituted, it is substituted with at least one selected from the group consisting of methyl, methoxy, halogen and hydroxy, n is 2, a quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof.

3. In paragraph 2, In the above [chemical formula 1], C3-C of A 20 The heterocycloalkyl is either morpholine or piperidine, A quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof.

4. In paragraph 1, A quinazoline-benzimidazole derivative represented by the above [chemical formula 1] is characterized in that it is one selected from the group consisting of the following compounds, or a pharmaceutically acceptable salt thereof: [Chemical Formula 1-1] [Chemical Formula 1-2] [Chemical Formula 1-3] [Chemical Formula 1-4] [Chemical Formula 1-5] [Chemical Formula 1-6] [Chemical Formula 1-7] [Chemical Formula 1-8] [Chemical Formula 1-9] [Chemical Formula 1-10] [Chemical Formula 1-11] [Chemical Formula 1-12] [Chemical Formula 1-13] [Chemical Formula 1-14] [Chemical Formula 1-15] [Chemical Formula 1-16] [Chemical Formula 1-17] [Chemical Formula 1-18] [Chemical Formula 1-19] [Chemical Formula 1-20] [Chemical Formula 1-21] [Chemical Formula 1-22] [Chemical Formula 1-23] [Chemical Formula 1-24] [Chemical Formula 1-25] [Chemical Formula 1-26] [Chemical Formula 1-27] [Chemical Formula 1-28] [Chemical Formula 1-29] [Chemical Formula 1-30] [Chemical Formula 1-31] 5. In paragraph 1, A quinazoline-benzimidazole derivative represented by the above [chemical formula 1] is characterized in that it is one selected from the group consisting of the following compounds, or a pharmaceutically acceptable salt thereof: [Chemical Formula 1-9] [Chemical Formula 1-10] [Chemical Formula 1-11] [Chemical Formula 1-13] [Chemical Formula 1-15] [Chemical Formula 1-16] [Chemical Formula 1-17] [Chemical Formula 1-20] [Chemical Formula 1-22] 6. In paragraph 1, A quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof, characterized in that the above quinazoline-benzimidazole derivative has both an effect of inhibiting tubulin polymerization and an effect of inhibiting tyrosine kinase of human epidermal growth receptor 2 (HER2).

7. In paragraph 1, A quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof, characterized in that the pharmaceutically acceptable salt of the quinazoline-benzimidazole derivative is at least one selected from the group consisting of hydrochloride, bromate, sulfate, phosphate, nitrate, citrate, acetate, lactate, tartrate, maleate, gluconate, succinate, formate, trifluoroacetate, oxalate, fumarate, glutarate, adipate, methanesulfonate, benzenesulfonate, paratoluenesulfonate, camphorsulfonate, sodium salt, potassium salt, lithium salt, calcium salt, and magnesium salt.

8. A pharmaceutical composition for preventing or treating cancer, comprising a quinazoline-benzimidazole derivative according to any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, as an active ingredient.

9. In paragraph 8, A pharmaceutical composition for preventing or treating cancer, characterized in that the pharmaceutical composition induces apoptosis by arresting the cancer cell cycle.

10. In paragraph 8, The above pharmaceutical composition is a pharmaceutical composition for preventing or treating cancer, which inhibits the activity of any one or more of HER2, p95HER2, HER3, EGFR, AKT, and MEK.

11. In paragraph 8, A pharmaceutical composition for preventing or treating cancer, characterized in that the cancer is at least one selected from the group consisting of skin cancer, breast cancer, uterine cancer, esophageal cancer, stomach cancer, brain tumor, colon cancer, rectal cancer, large intestine cancer, lung cancer, ovarian cancer, cervical cancer, endometrial cancer, vulvar cancer, kidney cancer, blood cancer, pancreatic cancer, prostate cancer, testicular cancer, laryngeal cancer, head and neck cancer, thyroid cancer, liver cancer, bladder cancer, osteosarcoma, lymphoma, blood cancer, thymic cancer, urethral cancer, and bronchial cancer.

12. In paragraph 8, A pharmaceutical composition for preventing or treating cancer, characterized in that the cancer is HER2 positive breast cancer or triple negative breast cancer.

13. A method for producing a quinazoline-benzimidazole derivative represented by the following [chemical formula 1] or a pharmaceutically acceptable salt thereof, characterized in that it is produced through the following production steps: 1) R 1 A step of preparing a compound represented by the following [chemical formula 4] by reacting a compound represented by -NH2 with 5-bromoanthranilic acid and triethyl orthoformate; 2) A step of preparing a compound represented by [Chemical Formula 2] by reacting a compound represented by [Chemical Formula 4] with either 2-nitro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline or 4-amino-3-nitrophenylboronic acid pinacol ester; 3) A step of preparing a compound represented by [Chemical Formula 3] from a compound represented by [Chemical Formula 2]; and 4) A compound represented by [chemical formula 3] or R 2 A step of producing a compound represented by [chemical formula 1] by reacting with any one of the compounds represented by -CHO; [Chemical Formula 1] [Chemical Formula 2] [Chemical Formula 3] [Chemical Formula 4] In the above chemical formulas 1 to 4, R 1 Silver -(CH2) n -A and, A is substituted or unsubstituted C3-C 20 Cycloalkyl or C3-C 20 Any one of the heterocycloalkyls, When the above cycloalkyl or heterocycloalkyl is substituted, it is substituted with at least one selected from the group consisting of a C1-C6 alkyl group, a hydroxyl group, an amine group, a methoxy group, and a halogen group, R 2 is substituted or unsubstituted C3-C 20 Aryl, substituted or unsubstituted C3-C 20 Heteroaryl, substituted or unsubstituted C3-C 20 Cycloalkyl, substituted or unsubstituted C3-C 20 Any one selected from the group consisting of heterocycloalkyl and methyl carbamate, When the above aryl, cycloalkyl or heterocycloalkyl is substituted, it is substituted with at least one selected from the group consisting of a C1-C6 alkyl group, a halogen group, a hydroxy group and an alkoxy group, and n is an integer from 0 to 6.

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