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 for preventing or treating cancer comprising the derivative as an active ingredient, the quinazoline-benzimidazole derivative of the present invention being activated in cancer cells, the compound can inhibit tubulin polymerization and tyrosine kinase of a human epidermal growth factor receptor 2 (HER2), can cause cancer cell cycle arrest to induce cell apoptosis and hinder cell growth signals transmitted through the HER2 receptor when being administered to a subject, can be used as a novel double-targeting anti-cancer drug, can prevent or treat cancers, and can be used as a novel anti-cancer drug. Preferably, HER2 positive breast cancer or triple negative breast cancer is prevented or treated.

Description

TECHNICAL FIELD

[0001] The present application relates to quinazoline-benzimidazole derivatives or pharmaceutically acceptable salts thereof, and uses thereof, and the like. BACKGROUND

[0002] Triple-negative breast cancer (TNBC, ER-, PR-, HER2-) patients account for 10% to 15% of all breast cancer patients, and the patients cannot benefit from hormone therapy or HER2 targeted therapy due to the lack of hormone receptors (ER (estrogen receptor), PR (progesterone receptor)) and HER2 protein. At present, the standard treatment of triple-negative breast cancer completely relies on conventional cytotoxic anticancer drugs (Taxene or Anthracycline). Since there is no clear targeted therapeutic drug, the treatment strategy for other subtypes is very single compared with breast cancer. More seriously, most patients will relapse within 2 to 3 years after surgery or chemotherapy, and are also prone to induce lung, liver, brain, bone and other organ metastases, thereby reducing the survival rate of patients.

[0003] The 5-year overall survival of patients diagnosed as stage-III is less than 55%, and the 5-year overall survival of patients with advanced-stage is less than 30%, which is at a very low level. The disease is a very serious disease that most patients will die within a few years.

[0004] Microtubules are the main components of the cytoskeleton, which are composed of heteromultimers of tubulin based on α and β subunits. Microtubules can achieve many cell functions, such as intracellular transport, polarity maintenance, intracellular signal transduction, cell migration and proliferation, etc. During the mitotic phase, microtubules form spindle fibers to separate chromosomes arranged in the cell center to two poles. If the spindle fibers cannot normally function, cell division will be inhibited to induce apoptosis, so it is an attractive target for anticancer drugs.

[0005] Drugs targeting microtubules can be broadly categorized into those that stabilize microtubules and those that disrupt their stability. First, microtubule stabilizers include taxane, paclitaxel (Taxol), and decetaxel, which prevent microtubule depolymerization and promote polymerization. Most microtubule stabilizers bind to taxane binding sites or overlapping sites of β-tubulin. Second, microtubule destabilizers include colchicine and vinca alkaloids, which bind to colchicine binding sites or vinca binding sites. Compared to drugs that affect microtubule polymerization, drugs targeting the microtubules themselves can be effective at lower drug concentrations, ultimately inhibiting cell mitosis. Therefore, there is a need to develop potential microtubule polymerization inhibitors as anticancer drugs.

[0006] As is well known, human epidermal growth factor receptor 2 (HER2) is overexpressed in various cancers, especially breast and gastric cancer, and is one of the important factors promoting cancer cell growth, survival, differentiation, and metastasis. HER2 activity activates intracellular signaling pathways through the receptor's tyrosine kinase domain. These pathways mainly include the RAS / RAF / MEK / ERK and PI3K / AKT / mTOR pathways. These signaling pathways can induce rapid proliferation of cancer cells and induce inhibition of apoptosis, thereby promoting tumor formation and progression.

[0007] Existing research has developed various compounds that block this signaling pathway by inhibiting the tyrosine kinase activity of HER2, effectively suppressing the growth and metastasis of cancer cells. Representative HER2 tyrosine kinase inhibitors include lapatinib and neratinib, which bind to the ATP-binding site of the HER2 receptor, thereby inhibiting signal transduction by blocking the autophosphorylation of tyrosine residues. These drugs have been used as effective treatments, particularly for patients with HER2-positive breast cancer. However, due to the emergence of drug resistance and side effects, the need to develop novel HER2 tyrosine kinase inhibitors remains a focus of attention. Summary of the Invention

[0008] Technical problems to be solved

[0009] The purpose of this invention is to provide a quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof.

[0010] Furthermore, another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of cancer, comprising the quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof as an active ingredient.

[0011] Another object of the present invention is to provide a method for preparing the quinazoline-benzimidazole derivative or its pharmaceutically acceptable salt.

[0012] However, the technical problem to be solved by the present invention is not limited to the above-mentioned problems, and other problems not mentioned will be clearly understood by those skilled in the art through the following description.

[0013] Technical methods for solving problems

[0014] To address the above problems, the present invention provides a quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof represented by the following [Formula 1]: [Formula 1] ; In Equation 1, R 1 It is -(CH2) n -A, A is C3-C, whether substituted or unsubstituted. 20 cycloalkyl or C3-C 20 Any of the heterocyclic alkyl groups, If the cycloalkyl or heterocycloalkyl group is substituted, it is by any one or more substitutions selected from the group consisting of C1-C6 alkyl, hydroxyl, amino, methoxy, and halogen groups. R 2 Choose C3-C, whether substituted or unsubstituted. 20 Aryl, substituted or unsubstituted C3-C 20 heteroaryl, substituted or unsubstituted C3-C 20 cycloalkyl, substituted or unsubstituted C3-C 20 Any one of the group consisting of heterocyclic alkyl groups and methyl carbamates. If the aryl, cycloalkyl, or heterocycloalkyl group is substituted, it is substituted by one or more of the group consisting of C1-C6 alkyl, halogen, hydroxyl, and alkoxy groups, where n is an integer from 0 to 6.

[0015] According to one aspect, In the above [Equation 1], A is C3-C, whether substituted or unsubstituted. 20 Heterocyclic alkyl groups If the heterocyclic alkyl group is substituted, it is substituted by one or more of the methyl or hydroxyl groups. R2 It is selected from any one of the group consisting of substituted or unsubstituted phenyl, substituted or unsubstituted thiophene, substituted or unsubstituted pyridyl, substituted or unsubstituted cyclopentyl, substituted or unsubstituted cyclohexyl and -NH-(C=O)-OCH3. If the phenyl, thiophene, pyridyl, cyclopentyl, and cyclohexyl groups are substituted, it is by any one or more substitutions selected from the group consisting of methyl, methoxy, halogen, and hydroxyl groups, where n is 2.

[0016] According to one aspect, In the above [Equation 1], A's C3-C 20 The heterocyclic alkyl group is either morpholine or piperidine.

[0017] According to one aspect, The quinazoline-benzimidazole derivative represented by [Formula 1] is selected from any one of the following groups of compounds: [Equation 1-1] ; [Equation 1-2] ; [Equation 1-3] ; [Equations 1-4] ; [Equations 1-5] ; [Equations 1-6] ; [Equations 1-7] ; [Equations 1-8] ; [Equations 1-9] ; [Equation 1-10] ; [Equation 1-11] ; [Equation 1-12] ; [Equation 1-13] ; [Equation 1-14] ; [Equation 1-15] ; [Equation 1-16] ; [Equation 1-17] ; [Equation 1-18] ; [Equation 1-19] ; [Equation 1-20] ; [Equation 1-21] ; [Equation 1-22] ; [Equation 1-23] ; [Equation 1-24] ; [Equation 1-25] ; [Equation 1-26] ; [Equation 1-27] ; [Equation 1-28] ; [Equation 1-29] ; [Equation 1-30] ; [Equation 1-31] .

[0018] According to one aspect, the quinazoline-benzimidazole derivative has both the effect of inhibiting tubulin polymerization and the effect of inhibiting human epidermal growth factor receptor 2 (HER2) tyrosine kinase.

[0019] According to one aspect, the pharmaceutically acceptable salt of the quinazoline-benzimidazole derivative is selected from any one or more of the group consisting of hydrochloride, bromate, sulfate, phosphate, nitrate, citrate, acetate, lactate, tartrate, maleate, gluconate, succinate, formate, trifluoroacetate, oxalate, fumarate, glutarate, adipate, methanesulfonate, benzenesulfonate, p-toluenesulfonate, camphorsulfonate, sodium salt, potassium salt, lithium salt, calcium salt, and magnesium salt.

[0020] According to another embodiment of the present invention, a pharmaceutical composition for the prevention or treatment of cancer is provided, comprising the quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof as an active ingredient.

[0021] According to one aspect, the pharmaceutical composition can induce apoptosis by achieving cell cycle arrest in cancer cells.

[0022] According to one aspect, the pharmaceutical composition can inhibit the activity of any one or more of HER2, p95HER2, HER3, EGFR, AKT, and MEK.

[0023] According to one aspect, the cancer is selected from one or more of the group consisting of skin cancer, breast cancer, uterine cancer, esophageal cancer, gastric cancer, brain tumor, colon cancer, rectal cancer, colorectal cancer, lung cancer, ovarian cancer, cervical cancer, endometrial cancer, vulvar cancer, kidney cancer, hematologic malignancies, pancreatic cancer, prostate cancer, testicular cancer, laryngeal cancer, head and neck cancer, thyroid cancer, liver cancer, bladder cancer, osteosarcoma, lymphoma, hematologic malignancies, thymic cancer, urethral cancer, and bronchial cancer, preferably HER2-positive breast cancer or triple-negative breast cancer.

[0024] According to another embodiment of the present invention, a method for preparing a quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof represented by the following [Formula 1] is provided, characterized by preparation by the following steps: 1) By making R 1 The steps for preparing the compound represented by the following [Formula 4] by reacting the compound represented by -NH2 with 5-bromoanthranilic acid and triethyl orthoformate; 2) The step of preparing the compound represented by [Formula 2] by reacting the compound represented by [Formula 4] with either 2-nitro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaneborane-2-yl)aniline or 4-amino-3-nitrophenylboronic acid pinacol ester; 3) The steps for preparing the compound represented by [Formula 3] from the compound represented by [Formula 2]; and 4) By reacting the compound represented by [Equation 3] with... The compound represented by R or derived from R 2 The steps for preparing the compound represented by [Formula 1] by reacting any of the compounds represented by -CHO; [Formula 1] ; [Equation 2] ; [Formula 3] ; [Formula 4] ; In equations 1 to 4, R 1 It is -(CH2) n -A, A is C3-C, whether substituted or unsubstituted. 20 cycloalkyl or C3-C 20 Any of the heterocyclic alkyl groups, If the cycloalkyl or heterocycloalkyl group is substituted, it is by any one or more substitutions selected from the group consisting of C1-C6 alkyl, hydroxyl, amino, methoxy, and halogen groups. R 2 Choose C3-C, whether substituted or unsubstituted. 20 Aryl, substituted or unsubstituted C3-C 20 heteroaryl, substituted or unsubstituted C3-C 20 cycloalkyl, substituted or unsubstituted C3-C 20 Any one of the group consisting of heterocyclic alkyl groups and methyl carbamates. If the aryl, cycloalkyl, or heterocycloalkyl group is substituted, it is substituted by one or more of the group consisting of C1-C6 alkyl, halogen, hydroxyl, and alkoxy groups, where n is an integer from 0 to 6.

[0025] In addition, the present invention provides a composition for the prevention or treatment of cancer comprising the quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof as an active ingredient.

[0026] In addition, the present invention provides a method for preventing or treating cancer, comprising the step of administering the quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof to a subject.

[0027] In addition, the present invention provides the use of the quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the prevention or treatment of cancer.

[0028] In addition, the present invention provides a cancer diagnosis method comprising the step of administering the quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof to a subject.

[0029] In addition, the present invention provides the use of the quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof in the preparation of a medicament for diagnosing cancer.

[0030] According to one embodiment of the present invention, the pharmaceutical composition can induce cell cycle arrest in cancer cells, thereby inducing apoptosis, or it can block cell growth signals transmitted through the HER2 receptor by inhibiting the tyrosine kinase of human epidermal growth factor receptor 2 (HER2). Preferably, both of the above mechanisms can be achieved.

[0031] According to another embodiment of the present invention, the cancer is selected from one or more of the group consisting of skin cancer, breast cancer, uterine cancer, esophageal cancer, gastric cancer, brain tumor, colon cancer, rectal cancer, colorectal cancer, lung cancer, ovarian cancer, cervical cancer, endometrial cancer, vulvar cancer, kidney cancer, hematologic malignancies, 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, preferably breast cancer, more preferably HER2-positive breast cancer or triple-negative breast cancer.

[0032] Invention Effects

[0033] This invention relates to a quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof, and compositions containing said derivative as an active ingredient for the prevention or treatment of cancer. The quinazoline-benzimidazole derivative of this invention is activated in cancer cells and can inhibit human epidermal growth factor receptor 2 (HER2) tyrosine kinase while inhibiting tubulin polymerization. Therefore, upon administration to a subject, it can block the cancer cell cycle and induce apoptosis, and block cell growth signals transmitted through the HER2 receptor, thereby providing an anti-cancer effect while inhibiting cell proliferation and survival. Therefore, the compounds of this invention and compositions containing them, as novel dual-target anti-cancer compositions, can be used for the prevention or treatment of cancer, preferably for the prevention or treatment of HER2-positive or triple-negative breast cancer. Attached Figure Description

[0034] Figure 1The results of the MTS assay were shown after treating triple-negative breast cancer cell line MDA-MB-231 and HER2-positive breast cancer cell line JIMT-1 with different concentrations (0 μM, 1 μM and 5 μM) of quinazoline-benzimidazole derivatives (1-1 to 1-10).

[0035] Figure 2 The results of the MTS assay were shown after treating triple-negative breast cancer cell line MDA-MB-231 and HER2-positive breast cancer cell line JIMT-1 with different concentrations (0 μM, 1 μM and 5 μM) of quinazoline-benzimidazole derivatives (1-11 to 1-20).

[0036] Figure 3 The results of the MTS assay were shown after treating triple-negative breast cancer cell line MDA-MB-231 and HER2-positive breast cancer cell line JIMT-1 with different concentrations (0 μM, 1 μM and 5 μM) of quinazoline-benzimidazole derivatives (1-21 to 1-31).

[0037] Figure 4 The cell viability and apoptosis morphology of HER2-positive breast cancer cell lines sensitive and resistant to trastuzumab are shown.

[0038] Figure 5 The results show that microtubule degradation and apoptosis were confirmed in SKBR3 and JIMT-1 cell lines.

[0039] Figure 6 The results show the confirmation of cell cycle arrest and the expression of mitotic markers in the G2 / M phase.

[0040] Figure 7 Western blot analysis results showing the expression and activity of HER2, HER family members and downstream signaling pathway factors.

[0041] Figure 8 The effects of quinazoline-benzimidazole derivative compounds 1-16 and 1-19 on lung cancer, colorectal cancer, ovarian cancer, prostate cancer, liver cancer, and hematologic malignancies were demonstrated.

[0042] The best form of implementing an invention

[0043] The inventors have demonstrated that benzimidazole derivatives can achieve cell cycle arrest by inhibiting tubulin synthesis, thus exhibiting an anticancer effect by inhibiting cancer cell division. They have also confirmed that quinazoline derivatives can exert anticancer effects by inhibiting human epidermal growth factor receptor 2 (HER2) tyrosine kinase. Furthermore, the inventors have demonstrated that combining benzimidazole and quinazoline in a single compound molecule can simultaneously activate both mechanisms of action. Therefore, they have invented a novel dual-target anticancer drug, a quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof.

[0044] Based on the above results, the present invention provides a quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof represented by the following [Formula 1]: [Formula 1] ; In Equation 1, R 1 It is -(CH2) n -A, A is C3-C, whether substituted or unsubstituted. 20 cycloalkyl or C3-C 20 Any of the heterocyclic alkyl groups, If the cycloalkyl or heterocycloalkyl group is substituted, it is by any one or more substitutions selected from the group consisting of C1-C6 alkyl, hydroxyl, amino, methoxy, and halogen groups. R 2 Choose C3-C, whether substituted or unsubstituted. 20 Aryl, substituted or unsubstituted C3-C 20 heteroaryl, substituted or unsubstituted C3-C 20 cycloalkyl, substituted or unsubstituted C3-C 20 Any one of the group consisting of heterocyclic alkyl groups and methyl carbamates. If the aryl, cycloalkyl, or heterocycloalkyl group is substituted, it is substituted by one or more of the group consisting of C1-C6 alkyl, halogen, hydroxyl, and alkoxy groups, where n is an integer from 0 to 6.

[0045] In this invention, the term "substitution" refers to a reaction in which one atom or group of atoms in a compound molecule is replaced by another atom or group of atoms.

[0046] In this invention, the term "chain-like" refers to a molecule with a chain-like structure. A chain-like structure is a chemical structure in which carbon atoms are connected in a chain-like manner, including straight chains and branched chains.

[0047] In this invention, the term "cyclic" refers to a ring structure formed by connecting the two ends of an organic compound skeleton.

[0048] In this invention, the term "straight-chain or branched alkyl" refers to a monovalent straight-chain or branched hydrocarbon residue having 1 to 6 carbon atoms and consisting only of carbon and hydrogen atoms.

[0049] In this invention, the term "cycloalkyl" refers to cyclic saturated hydrocarbon residues.

[0050] Examples of the alkyl group include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, 2-butyl, 3-butyl, pentyl, n-hexyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.

[0051] In this invention, the term "heterocyclic alkyl" generally refers to a saturated or unsaturated (but not aromatic) cyclic hydrocarbon, which may optionally be unsubstituted, monosubstituted or polysubstituted, and has at least one heteroatom selected from N, O or S in its structure.

[0052] In this invention, the term "aryl" refers to an unsaturated aromatic ring compound having 3 to 12 carbon atoms, which has a monocyclic ring (e.g., phenyl) or multiple fused rings (e.g., naphthyl). Examples of such aryl groups include, but are not limited to, phenyl, naphthyl, etc.

[0053] In this invention, the term "heteroaryl" refers to a monocyclic or fused ring in which at least one of the atoms constituting the ring is a heteroatom of N, O, or S. Examples of such heteroaryl groups include, but are not limited to, pyridinyl, pyrimidinyl, pyrazinyl, oxazolyl, furanyl, etc.

[0054] In this invention, the term "alkoxy" refers to an alkyl group (-OR) bonded to oxygen. Examples of such alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, etc.

[0055] In this invention, the "halogen group" can be fluorine (F), chlorine (Cl), bromine (Br) or iodine (I), etc.

[0056] In a preferred embodiment of the invention, the compound represented by Formula 1 is preferably one or more selected from the group consisting of the following compounds: [Equation 1-1] ; [Equation 1-2] ; [Equation 1-3] ; [Equations 1-4] ; [Equations 1-5] ; [Equations 1-6] ; [Equations 1-7] ; [Equations 1-8] ; [Equations 1-9] ; [Equation 1-10] ; [Equation 1-11] ; [Equation 1-12] ; [Equation 1-13] ; [Equation 1-14] ; [Equation 1-15] ; [Equation 1-16] ; [Equation 1-17] ; [Equation 1-18] ; [Equation 1-19] ; [Equation 1-20] ; [Equation 1-21] ; [Equation 1-22] ; [Equation 1-23] ; [Equation 1-24] ; [Equation 1-25] ; [Equation 1-26] ; [Equation 1-27] ; [Equation 1-28] ; [Equation 1-29] ; [Equation 1-30] ; [Equation 1-31] .

[0057] The quinazoline-benzimidazole derivative or its pharmaceutically acceptable salt described in this invention has specific activity in cancer cells, inhibiting microtubule polymerization and inducing apoptosis. In addition, it can inhibit human epidermal growth factor receptor 2 (HER2) tyrosine kinase, thereby blocking cell growth signals transmitted through the HER2 receptor and inhibiting the activity of HER2, p95HER2, HER3, EGFR, AKT and MEK, etc. Therefore, it can be included as an active ingredient in pharmaceutical compositions for the prevention or treatment of cancer.

[0058] In one embodiment of the present invention, the pharmaceutical composition can induce apoptosis by achieving cell cycle arrest in cancer cells, or can block cell growth signals transmitted through the HER2 receptor. Preferably, both of these mechanisms of action can be achieved.

[0059] In this invention, the term "pharmaceutically acceptable salt" refers to a dosage form of the compound that will not cause severe irritation to the organism administering the compound and will not impair the compound's biological activity and physical properties. Pharmaceutically acceptable salts can be prepared by reacting the compounds of this invention with inorganic acids such as hydrochloric acid, bromic acid, sulfuric acid, nitric acid, and phosphoric acid; sulfonic acids such as methanesulfonic acid, ethanesulfonic acid, and p-toluenesulfonic acid; and organic carboxylic acids such as tartaric acid, formic acid, citric acid, acetic acid, trichloroacetic acid, trifluoroacetic acid, decanoic acid, isobutyric acid, malonic acid, succinic acid, phthalic acid, gluconic acid, benzoic acid, lactic acid, fumaric acid, maleic acid, and salicylic acid. Furthermore, the compounds of this invention can be prepared by reacting them with a base to form salts of alkali metals such as ammonium, sodium, and potassium salts; salts of alkaline earth metals such as calcium and magnesium salts; salts of organic bases such as dicyclohexylamine, N-methyl-D-glucosamine, and tris(hydroxymethyl)methylamine; and salts of amino acids such as arginine and lysine.

[0060] Furthermore, the quinazoline-benzimidazole derivative or its pharmaceutically acceptable salts include not only pharmaceutically acceptable salts, but also all salts, hydrates, and solvates that can be prepared by conventional methods.

[0061] Furthermore, the present invention can provide a method for the prevention, treatment and / or diagnosis of cancer, comprising the step of administering the quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof to a subject.

[0062] In this invention, the term "prevention" refers to all actions that inhibit or delay the occurrence, spread, or recurrence of cancer by applying the composition of this invention; "treatment" refers to all actions that improve or benefit cancer symptoms by applying the composition of this invention.

[0063] In this invention, the term "pharmaceutical composition" is intended for the prevention or treatment of diseases and can be formulated into various forms for use according to conventional methods. For example, it can be formulated into oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, and syrups, or into external preparations, suppositories, and sterile injections.

[0064] In this invention, "comprising as an active ingredient" means that the content of the ingredient is sufficient or necessary to achieve the desired biological effect. In practical applications, the content of the active ingredient is determined according to the purpose of treating the disease and should not cause other toxicities. For example, it can be varied based on various factors such as the disease or condition being treated, the type of composition being administered, the size of the subject, or the severity of the disease or condition. Those skilled in the art to which this invention pertains can determine the effective content of a single composition empirically without requiring extensive experimentation.

[0065] Furthermore, in addition to the active ingredients of the above-mentioned dosage forms, the pharmaceutical compositions of the present invention may also contain one or more pharmaceutically acceptable carriers.

[0066] The pharmaceutically acceptable carrier may be saline, sterile water, Ringer's solution, buffered saline, glucose solution, maltodextrin solution, glycerol, ethanol, and at least one mixture of these components, and may further include, as needed, other conventional additives such as antioxidants, buffers, and antibacterial agents. Furthermore, diluents, dispersants, surfactants, binders, and lubricants may be added to formulate aqueous solutions, suspensions, emulsions, or other injectable preparations, pills, capsules, granules, or tablets. Additionally, formulation may preferably be carried out using appropriate methods in the art or methods disclosed in Remington's Pharmaceutical Science (Mack Publishing Company, Easton PA) to suit different diseases or ingredients.

[0067] The compositions of the present invention can be administered orally or parenterally in pharmaceutically effective amounts according to desired methods. The term "pharmaceutically effective amount" in the present invention refers to a content that can adequately treat a disease without causing side effects within a reasonable benefit / risk ratio of applicable drug treatment. The level of the effective amount can be determined based on many factors as well as other factors known in the medical field. Factors to be considered may include the patient's health status, disease severity, drug activity, drug sensitivity, method of administration, time of administration, route of administration and excretion rate, treatment duration, and drugs taken in combination or concurrently.

[0068] Furthermore, the pharmaceutical compositions of the present invention can be administered to subjects to prevent, treat, and / or diagnose cancers, including skin cancer, breast cancer, uterine cancer, esophageal cancer, gastric cancer, brain tumors, colon cancer, rectal cancer, colorectal cancer, lung cancer, ovarian cancer, cervical cancer, endometrial cancer, vulvar cancer, kidney cancer, hematologic malignancies, pancreatic cancer, prostate cancer, testicular cancer, laryngeal cancer, head and neck cancer, thyroid cancer, liver cancer, bladder cancer, osteosarcoma, lymphoma, hematologic malignancies, thymic cancer, urethral cancer, or bronchial cancer, preferably cancers with acidity higher than normal cells and whose cytotoxicity can be inhibited by microtubule polymerization inhibitors. Non-limiting examples include breast cancer, preferably HER2-positive breast cancer or triple-negative breast cancer.

[0069] In this invention, the term "subject" can be any mammal, such as livestock or humans, that requires prevention, treatment, and / or diagnosis of cancer, without particular limitation, but is preferably human.

[0070] The pharmaceutical compositions of the present invention can be formulated into various dosage forms for administration to subjects. A representative parenteral dosage form is an injectable dosage form, preferably an isotonic aqueous solution or suspension. Injectable dosage forms can be prepared using suitable dispersants or wetting agents and suspending agents according to techniques known in the art. For example, the components can be dissolved in physiological saline or buffer solution to prepare an injection. Furthermore, dosage forms for oral administration include ingestible tablets, sublingual tablets, lozenges, capsules, elixirs, suspensions, syrups, and rice paper capsules, etc. These dosage forms, in addition to the active ingredient, may include diluents (e.g., lactose, glucose, sucrose, mannitol, sorbitol, cellulose, and / or glycine) and lubricants (e.g., silica, talc, stearic acid and its magnesium or calcium salts, and / or polyethylene glycol). The tablets may contain binders such as magnesium aluminum silicate, starch paste, gelatin, astragalus gum, methylcellulose, sodium carboxymethyl cellulose, and / or polyvinylpyrrolidone, and may further include disintegrants, absorbents, colorants, flavoring agents, and / or sweeteners such as starch, agar, alginate, or their sodium salts. The formulation can be prepared by conventional mixing, granulation, or coating methods.

[0071] Furthermore, the pharmaceutical composition of the present invention may also include preservatives, hydrating agents, emulsification promoters, salts or buffers for controlling osmotic pressure, and other substances that can be used for treatment, and may be formulated according to conventional methods.

[0072] The pharmaceutical compositions of the present invention can be administered via various routes, including oral, transdermal, subcutaneous, intravenous, or intramuscular administration. The appropriate dosage of the active ingredient can be determined based on factors such as the route of administration, the patient's age, sex, weight, and disease severity. Furthermore, the compositions of the present invention can be used in combination with known compounds that can enhance the desired effect.

[0073] The pharmaceutical compositions according to the present invention can be administered to humans or animals via oral administration or parenteral administration, such as intravenous, subcutaneous, intranasal, or intraperitoneal administration. Oral administration also includes sublingual administration. Parenteral administration includes injection methods such as subcutaneous injection, intramuscular injection, and intravenous injection, as well as infusion methods.

[0074] In the pharmaceutical compositions of the present invention, the total effective amount of the quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof can be administered to the patient via a single dose or via a fractionated treatment protocol with multiple doses over a prolonged period. The content of the active ingredient in the pharmaceutical compositions of the present invention can be varied according to the severity of the disease, but generally, based on an adult, the effective dose per administration can be from 100 μg to 3000 mg, administered multiple times a day. However, the concentration of the quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof depends on various factors, such as the patient's age, weight, health status, sex, disease severity, diet, and excretion rate, and these factors can be taken into account when determining the patient's dosage.

[0075] Furthermore, as long as the pharmaceutical composition of the present invention achieves the effects of the present invention, there are no particular limitations on its dosage form, route of administration, or method of administration. In addition to the quinazoline-benzimidazole derivative or its pharmaceutically acceptable salt as the active ingredient, the pharmaceutical composition of the present invention may further include known anticancer agents and may be used in combination with other known treatment methods.

[0076] The terminology used in the embodiments is for illustrative purposes only and is not intended to limit the scope. Unless the context clearly indicates that they have different meanings, the singular form encompasses the plural form. In this specification, terms such as "comprising" or "having" are used to express the presence of the features, numbers, steps, operations, constituent elements, accessories, or combinations thereof described in the specification, and do not exclude the possibility of the presence or additional addition of one or more other features, numbers, steps, operations, constituent elements, accessories, or combinations thereof.

[0077] Unless otherwise defined, all terms used herein, including technical or scientific terms, shall have their ordinary meanings as understood by one of ordinary skill in the art. Terms commonly used, such as those defined in dictionaries, shall be understood as having their meanings in the relevant technical context and shall not be interpreted as idealized or overly formalized meanings unless explicitly defined in this specification.

[0078] The exemplary embodiments will now be described in detail with reference to the accompanying drawings. However, various modifications can be made to the exemplary embodiments. This disclosure is not limited to the exemplary embodiments herein. The exemplary embodiments should be understood to include all modifications, equivalents, and alternatives thereof made within the concept and scope of this disclosure.

[0079] Furthermore, the embodiments disclosed in this specification are described in detail with reference to the accompanying drawings. Identical or similar components are assigned the same reference numerals, regardless of the reference numerals, and repetitive descriptions are omitted. Additionally, when describing the embodiments disclosed in this specification, detailed descriptions of relevant prior art will be omitted if it is determined that such detailed descriptions may obscure the main idea of ​​the embodiments disclosed in this specification.

[0080] [Example]

[0081] Example 1. Preparation of quinazoline-benzimidazole derivatives

[0082] All chemical reagents are commercially available products. 1 The H NMR spectra were recorded on a Bruker Avance III 400 MHz instrument with TMS as an internal standard.

[0083] 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 formula 1-1)

[0084] [Step 1] Synthesis of 6-Bromo-3-(2-morpholinoethyl)quinazolin-4(3H)-one

[0085] 5-Bromoanthranilic acid (2.16 g, 9.3 mmol) was added to 30 mL of anhydrous ethanol and dissolved under a nitrogen atmosphere. Triethylorthoformate (1.39 g, 13 mmol) and 2-morpholinoethylamine (1.69 g, 13 mmol) were then added. Iodine (25 mg, 0.1 mmol) was added as a catalyst. The mixture was then heated under reflux for 6 hours. After the reaction was confirmed by TLC, the mixture was cooled to room temperature. Ethanol was removed by vacuum distillation, and the solution was then dissolved in 60 mL of ethyl acetate. The solution was washed with 1N-NaOH aqueous solution and pure water, dried over anhydrous Na₂SO₄, and filtered. The filtrate was then distilled under vacuum to give a bright yellow solid (2.21 g).

[0086] Yield: 69.7%

[0087] 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)

[0088] [Step 2] Synthesis of 6-(4-Amino-3-nitrophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (Formula 2-1)

[0089] 2-Nitro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (1 g, 3.8 mmol), 6-bromo-3-(2-morpholinoethyl)quinazolin-4(3H)-one (1.28 g, 3.8 mmol), and K₂CO₃ (1.56 g, 11.4 mmol) were dissolved in 25 mL of dioxane under a nitrogen atmosphere. The mixture was then heated to 80 °C, followed by the addition of Pd(dppf)Cl₂ (0.14 g, 0.19 mmol) and reflux for 6 hours. The reaction was confirmed by TLC until completion, at which point the mixture was distilled under reduced pressure. The solution was then purified by column chromatography to obtain a yellow solid (0.58 g).

[0090] Yield: 38.9%

[0091] 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)

[0092] [Step 3] Synthesis of 6-(3,4-Diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (Formula 3-1)

[0093] 6-(4-amino-3-nitrophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.58 g, 1.4 mmol) was dissolved in 20 mL of methanol, and 0.35 g of 10%-Pd / C was added with stirring at room temperature. Then, hydrazine hydrate (0.75 g, 14 mmol) was added dropwise, and the mixture was heated to reflux for 3 hours. After confirming the reaction by TLC, the mixture was cooled to room temperature. The solution was then filtered through celite and distilled under reduced pressure. The filtrate was then dissolved again in 20 mL of methylene chloride and dried over anhydrous MgSO4. The solution was filtered and distilled under reduced pressure to give a yellow solid (0.39 g).

[0094] Yield 73.5%, 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)

[0095] 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)

[0096] 6-(3,4-diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.2 g, 0.5 mmol) and 1,3-bis(methoxycarbonyl)-2-methyl-2-thiopseudourea (0.29 g, 14 mmol) were added to 15 mL of 5% acetic acid in ethanol solution (5%-AcOH in Ethanol), and the mixture was heated under reflux for 3 hours. After precipitation, the mixture was cooled and filtered to obtain a yellow solid (0.17 g).

[0097] Yield 73%

[0098] 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)

[0099] 1-2. Synthesis of 3-(2-Morpholinoethyl)-6-(2-(p-tolyl)-1H-benzo[d]imidazol-6-yl)quinazolin-4(3H)-one (Formula 1-2)

[0100] 6-(3,4-diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.15 g, 0.4 mmol) and p-methylbenzaldehyde (50 mg, 0.4 mmol) 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, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. The resulting product was purified by column chromatography to give a pale yellow solid (0.1 g).

[0101] Yield: 52.53%

[0102] 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)

[0103] 1-3. Synthesis of 6-(2-(4-fluorophenyl)-1H-benzo[d]imidazol-6-yl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (Formula 1-3)

[0104] 6-(3,4-Diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.15 g, 0.4 mmol) and p-fluorobenzaldehyde (50 mg, 0.4 mmol) 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, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. The resulting product was purified by column chromatography to obtain a pale yellow solid.

[0105] (0.07 g)

[0106] Yield 36.8%

[0107] 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)

[0108] Synthesis of 1-4,6-(2-(3-fluorophenyl)-1H-benzo[d]imidazol-6-yl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (Formula 1-4)

[0109] 6-(3,4-diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.15 g, 0.4 mmol) and m-fluorobenzaldehyde (50 mg, 0.4 mmol) 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, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. The resulting product was purified by column chromatography to obtain a pale yellow solid.

[0110] (0.048 g)

[0111] Yield 22.85%

[0112] 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)

[0113] 1-5. Synthesis of 6-(2-(4-Chlorophenyl)-1H-benzo[d]imidazol-6-yl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (Formula 1-5)

[0114] 6-(3,4-diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.15 g, 0.4 mmol) and p-chlorobenzaldehyde (58 mg, 0.4 mmol) 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, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. The resulting product was purified by column chromatography to give a pale yellow solid (0.075 g).

[0115] Yield 37.68%, 1H 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)

[0116] 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)

[0117] 6-(3,4-diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.15 g, 0.4 mmol) and 2,4-dihydroxybenzaldehyde (57 mg, 0.4 mmol) 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, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. The resulting product was purified by column chromatography to give a pale yellow solid (0.032 g).

[0118] Yield 16%

[0119] 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)

[0120] 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)

[0121] 6-(3,4-diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.15 g, 0.4 mmol) and 3-fluoro-6-hydroxybenzaldehyde (57 mg, 0.4 mmol) 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, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. The resulting product was purified by column chromatography to give a pale yellow solid (0.091 g).

[0122] Yield: 45.73%

[0123] 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)

[0124] 1-8. Synthesis of 6-(2-cyclohexyl-1H-benzo[d]imidazol-6-yl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (Formula 1-8)

[0125] 6-(3,4-diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.15 g, 0.4 mmol) and cyclohexylbenzaldehyde (46 mg, 0.4 mmol) 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, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. The resulting product was purified by column chromatography to give a pale yellow solid (0.030 g).

[0126] Yield 16%

[0127] 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)

[0128] 1-9. Synthesis of 6-(2-cyclopentyl-1H-benzo[d]imidazol-6-yl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (Formula 1-9)

[0129] 6-(3,4-diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.15 g, 0.4 mmol) and cyclopentylbenzaldehyde (40 mg, 0.4 mmol) 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, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. The resulting product was purified by column chromatography to give a pale yellow solid (0.024 g).

[0130] Yield 13.3%

[0131] 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=4.4Hz, 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)

[0132] 1-10. Synthesis of 6-(2-(4-methoxyphenyl)-1H-benzo[d]imidazoyl-3-yl-(2-morpholinoethyl)quinazolin-4(3H)-one (Formula 1-10)

[0133] 6-(3,4-diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.13 g, 0.35 mmol) and 2,4-difluorobenzaldehyde (48 mg, 0.35 mmol) 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, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. The resulting product was purified by column chromatography to give a pale yellow solid (0.070 g).

[0134] Yield: 41.17%

[0135] 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), 1-11. Synthesis of 6-(2-((3-Chloro-4-fluorophenyl)-1H-benzo[d]imidazoyl-3-(2-morpholinoethyl)quinazolin-4(3H)-one (Formula 1-11) 6-(3,4-diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.15 g, 0.4 mmol) and 3-chloro-4-fluorobenzaldehyde (65 mg, 0.4 mmol) 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, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. The resulting product was purified by column chromatography to give a pale yellow solid (0.116 g). Yield: 56.31% 1H 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)

[0136] 1-12. Synthesis of 6-(2-((2,4-Difluorophenyl)-1H-benzo[d]imidazoyl-3-yl-(2-morpholinoethyl)quinazolin-4(3H)-one (Formula 1-12)

[0137] 6-(3,4-diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.15 g, 0.4 mmol) and 2,4-difluorobenzaldehyde (58 mg, 0.4 mmol) 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, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. The resulting product was purified by column chromatography to give a pale yellow solid (0.057 g).

[0138] Yield 28.5%

[0139] 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)

[0140] 1-13. Synthesis of (6-(4-oxo-3-(2-(piperidin-1-yl)ethyl)-3,4-dihydroquinazolin-6-yl)-1H-benzo[d]imidazol-2-yl)carbamate (representative compound formula 1-13)

[0141] [Step 1] Synthesis of 6-bromo-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one

[0142] 30 mL of anhydrous ethanol was added to 2-amino-5-bromobenzoic acid (2.16 g, 9.4 mmol), followed by the sequential addition of 1-(2-aminoethyl)piperidine (1.56 g, 12.2 mmol), triethyl orthoformate (1.8 g, 12.2 mmol), and iodine (23 mg, 0.09 mmol) under a nitrogen atmosphere. The mixture was then heated to reflux for 6 hours. After confirming the reaction by TLC, the mixture was cooled to room temperature. All ethanol was removed by vacuum distillation, followed by the addition of 60 mL of ethyl acetate. The mixture was washed three times with 30 mL of 1N-NaOH and three times with 30 mL of brine. The organic layer was dried over anhydrous Na₂SO₄, filtered, and distilled under vacuum to obtain a brown liquid (2.43 g).

[0143] Yield: 77.14%

[0144] 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)

[0145] [Step 2] Synthesis of 6-(4-amino-3-nitrophenyl)-3-(2-(piperidin-1yl)ethyl)quinazolin-4(3H)-one (Formula 2-2)

[0146] 6-Bromo-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (1.0 g, 3.78 mmol), 4-Amino-3-nitrophenylboronic acid pinacol ester (1.27 g, 3.78 mmol), and potassium carbonate (1.56 g, 11.3 mmol) were dissolved in 25 mL of dioxane aqueous solution (20 mL dioxane + 5 mL pure water). The solution was heated to 100 °C, and then Pd(dppf)Cl2 (0.14 g, 0.18 mmol) catalyst was added. The mixture was heated under nitrogen atmosphere and refluxed for 6 hours. After confirmation of the reaction by TLC, the solution was filtered through diatomaceous earth to remove solid impurities. After adding 30 mL of ethyl acetate, the mixture was slurried for 1 hour and filtered to obtain a brown solid.

[0147] (0.68 g)

[0148] Yield 46%

[0149] 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)

[0150] [Step 3] Synthesis of 6-(3,4-Diaminophenyl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (Formula 3-2)

[0151] 6-(4-amino-3-nitrophenyl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.62 g, 1.58 mmol) was dissolved in 20 mL of methanol, and 0.37 g of 10%-Pd / C was added with stirring. Then, hydrazine hydrate (0.79 g, 15.8 mmol) was added, and the mixture was heated under reflux for 4 hours. After confirming the reaction by TLC, the mixture was filtered through diatomaceous earth. The organic layer was dried over anhydrous magnesium sulfate, filtered, and distilled under reduced pressure to give a bright yellow liquid.

[0152] (0.45 g)

[0153] Yield: 78.9%

[0154] 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)

[0155] [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)

[0156] 6-(3,4-diaminophenyl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.2 g, 0.5 mmol) and 1,3-bis(methoxycarbonyl)-2-methyl-2-thiopseudoureide (0.29 g, 14 mmol) were added to 15 mL of 5% acetic acid in ethanol solution (5%-AcOH in Ethanol) and heated under reflux for 3 hours. After precipitation, the solution was cooled and filtered to obtain a yellow solid (0.177 g).

[0157] Yield 72%

[0158] 1H 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=4.76Hz, 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), 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) 6-(3,4-diaminophenyl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.2 g, 0.55 mmol) and 4-fluorobenzaldehyde (68 mg, 0.55 mmol) 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, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. The resulting product was purified by column chromatography to give a pale yellow solid (0.080 g). Yield 42.1% 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), 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) 6-(3,4-diaminophenyl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.2 g, 0.55 mmol) and 4-chlorobenzaldehyde (77 mg, 0.55 mmol) 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, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. The resulting product was purified by column chromatography to give a pale yellow solid (0.103 g). Yield: 38.72% 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=4.76Hz, 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)

[0159] 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)

[0160] 6-(3,4-Diaminophenyl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.2 g, 0.55 mmol) and 3-chloro-4-fluorobenzaldehyde (87 mg, 0.55 mmol) were dissolved in 10 mL of DMF, and sodium metabisulfite (Na₂S₂O₅, 114 mg) was added. The mixture was stirred at 95 °C for 3 hours. After confirming the reaction by TLC, the mixture was cooled, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO₄, filtered, and concentrated under reduced pressure. Subsequently, the solution was purified by silica gel column chromatography using dichloromethane and methanol (5:1, v / v), dichloromethane and methanol (3:1, v / v), and ethyl acetate and methanol (5:1, v / v) as mixed eluents, yielding a pale yellow solid (0.048 g).

[0161] Yield: 17.39%

[0162] 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=4.76Hz, 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)

[0163] 1-17. Synthesis of 6-(2-(2,4-difluorophenyl)-1H-benzo[d]imidazol-6-yl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (Formula 1-17)

[0164] 6-(3,4-diaminophenyl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.2 g, 0.55 mmol) and 2,4-fluorobenzaldehyde (78 mg, 0.55 mmol) 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, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. The resulting product was purified by column chromatography to give a pale yellow solid (0.083 g).

[0165] Yield: 31.08%

[0166] 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=4.76Hz, 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)

[0167] 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)

[0168] 6-(3,4-diaminophenyl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.2 g, 0.55 mmol) and 4-methoxybenzaldehyde (75 mg, 0.55 mmol) 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, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. The resulting product was purified by column chromatography to give a pale yellow solid (0.084 g).

[0169] Yield: 31.93%

[0170] 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)

[0171] 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)

[0172] 6-(3,4-diaminophenyl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.15 g, 0.41 mmol) and 4-methylbenzaldehyde (49 mg, 0.41 mmol) 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, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. The resulting product was purified by column chromatography to give a pale yellow solid (0.080 g).

[0173] Yield 42.1%

[0174] 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)

[0175] 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)

[0176] 6-(3,4-diaminophenyl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.20 g, 0.55 mmol) and 3-thiophenecarboxaldehyde (62 mg, 0.55 mmol) 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, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. The resulting product was purified by column chromatography to give a pale yellow solid (0.10 g).

[0177] Yield 40.0%

[0178] 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)

[0179] 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)

[0180] 6-(3,4-diaminophenyl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.2 g, 0.55 mmol) and 4-pyridinecarboxaldehyde (59 mg, 0.55 mmol) 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, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. The resulting product was purified by column chromatography to give a pale yellow solid (0.03 g).

[0181] Yield 12.14%

[0182] 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)

[0183] 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)

[0184] 6-(3,4-diaminophenyl)-3-(2-(piperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.2 g, 0.55 mmol) and 5-chloro-2-pyridinecarboxaldehyde (78 mg, 0.55 mmol) 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, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. The resulting product was purified by column chromatography to give a pale yellow solid (0.148 g).

[0185] Yield: 55.63%

[0186] 1H 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)

[0187] 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)

[0188] 6-(3,4-diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.20 g, 0.55 mmol) and 3-thiophenecarboxaldehyde (61 mg, 0.55 mmol) 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, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. The resulting product was purified by column chromatography to give a pale yellow solid (0.15 g).

[0189] Yield 60.0%

[0190] 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)

[0191] 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)

[0192] 6-(3,4-diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.2 g, 0.55 mmol) and 4-pyridinecarboxaldehyde (58 mg, 0.55 mmol) 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, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. The resulting product was purified by column chromatography to give a pale yellow solid (0.02 g).

[0193] Yield 8.1%

[0194] 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)

[0195] 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)

[0196] 6-(3,4-Diaminophenyl)-3-(2-morpholinoethyl)quinazolin-4(3H)-one (0.22 g, 0.55 mmol) and 5-chloro-2-pyridinecarboxaldehyde (77 mg, 0.55 mmol) 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, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. The resulting product was purified by column chromatography to obtain a pale yellow solid.

[0197] (0.052 g)

[0198] Yield 19.54%

[0199] 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)

[0200] 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 formula 1-26)

[0201] [Step 1] Synthesis of 6-bromo-3-(2-(4-methylpiperidin-1-yl)ethyl)quinazolin-4(3H)-one

[0202] Under a nitrogen atmosphere, 2-amino-5-bromobenzoic acid (1.26 g, 5.5 mmol) and 2-(4-methyl-1-piperidinyl)ethanamine (1.0 g, 7.0 mmol) were added to 30 mL of anhydrous ethanol. Triethyl orthoformate (1.05 g, 7.0 mmol) and iodine (14 mg, 0.01 eq) were added, and the mixture was heated to reflux for 6 hours. After confirming the reaction by TLC, the mixture was slowly cooled. All solvent was then removed by vacuum distillation. The mixture was then diluted with 60 mL of ethyl acetate and washed with 1N-NaOH solution and brine. After drying with anhydrous Na₂SO₄, the mixture was filtered under reduced pressure. The filtrate was then distilled under reduced pressure to remove all residues, yielding a brown liquid.

[0203] (1.69 g)

[0204] Yield: 88.48%

[0205] 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=6Hz, 2H, -CH2), 2.83(m, 2H, -CH2), 4.06(t, J=6Hz, 2H, -CH2), 7.61~8.32(m, 4H,Aryl-H)

[0206] [Step 2] Synthesis of 6-(4-amino-3-nitrophenyl)-3-(2-(4-methylpiperidin-1-yl)ethyl)quinazolin-4(3H)-one (Formula 2-3)

[0207] After adding 2-nitro-4-(4,4,5,5-tetramethyl-1,3,2-diaxopentan-2-yl)aniline (1.27 g, 4.8 mmol), 6-bromo-3-(2-(4-methylpiperidin-1-yl)ethyl)quinazolin-4(3H)-one (1.68 g, 4.8 mmol) and K2CO3 (1.99 g, 14.4 mmol), 25 ml of wet dioxane was added and stirred under a nitrogen atmosphere.

[0208] Then, [1,1'-Bis(diphenylphosphino)ferrocene]palladium(II) dichloride (0.175 g, 0.05 eq, 0.175 g) was added, and the mixture was heated under reflux for 5 hours. After confirming the reaction by TLC, the mixture was slowly cooled. After cooling, the mixture was filtered through diatomaceous earth and the filtrate was distilled under reduced pressure. Ethyl acetate was then added, stirred, and filtered to give a brown solid.

[0209] (0.71 g)

[0210] Yield: 36.4%

[0211] 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=6Hz, 2H, -CH2), 2.87(m, 2H, -CH2), 4.09(t, J=6Hz, 2H, -CH2), 7.16~8.31(m, 7H,Aryl-H)

[0212] [Step 3] Synthesis of 6-(3,4-Diaminophenyl)-3-(2-(4-methylpiperidin-1-yl)ethyl)quinazolin-4(3H)-one (Formula 3-3)

[0213] 6-(4-amino-3-nitrophenyl)-3-(2-(4-methylpiperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.71 g, 1.74 mmol) was dissolved in 28 mL of methanol, and 0.35 g of 10%-Pd / C was added with stirring. Then, hydrazine hydrate (0.84 g, 17.4 mmol) was added, and the mixture was heated to reflux for 7 hours. After confirming the reaction by TLC, the mixture was cooled to room temperature. The solution was filtered through celite and distilled under reduced pressure. After dissolving in 20 mL of methylene chloride, the solution was dried over anhydrous magnesium sulfate. The solution was filtered and distilled under reduced pressure to give a brown liquid.

[0214] (0.27 g)

[0215] Yield 41.5%

[0216] 1 H NMR (400 MHz. DMSO-d6): δ 0.85(d, 3H, -CH3), 1.05(m, 2H, -CH2),1.29(m, 1H, C H -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)

[0217] [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)

[0218] 6-(3,4-diaminophenyl)-3-(2-(4-methylpiperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.135 g, 0.35 mmol) and 3-chloro-4-fluorobenzaldehyde (56 mg, 0.35 mmol) were dissolved in 10 mL of DMF, and sodium metabisulfite (Na₂S₂O₅, 85 mg) was added. The mixture was stirred at 95 °C for 3 hours. After confirming the reaction by TLC, the mixture was cooled, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO₄, filtered, and concentrated under reduced pressure. The resulting product was purified by column chromatography to obtain a pale yellow solid.

[0219] (55 mg)

[0220] Yield 30.5%

[0221] 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=6Hz, 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)

[0222] 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)

[0223] 6-(3,4-diaminophenyl)-3-(2-(4-methylpiperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.135 g, 0.35 mmol) and 4-chlorobenzaldehyde (50 mg, 0.35 mmol) were dissolved in 10 mL of DMF, and sodium metabisulfite (Na₂S₂O₅, 85 mg) was added. The mixture was stirred at 95 °C for 3 hours. After confirming the reaction by TLC, the mixture was cooled, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO₄, filtered, and concentrated under reduced pressure. The resulting product was purified by column chromatography to give a pale yellow solid (64 mg).

[0224] Yield 35.9%

[0225] 1 H NMR (400 MHz. DMSO-d6): δ 0.87(d, 3H, -CH3), 1.10(m, 2H, CH2),1.35(m, 1H, -C H -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)

[0226] 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 formula 1-28)

[0227] [Step 1] Synthesis of 6-bromo-3-(2-(4-hydroxypiperidin-1-yl)ethyl)quinazolin-4(3H)-one

[0228] Under a nitrogen atmosphere, 2-amino-5-bromobenzoic acid (1.26 g, 5.5 mmol) and 1-(2-aminoethyl)-4-piperidinol (1.0 g, 7.0 mmol) were added to 30 mL of anhydrous ethanol. Triethyl orthoformate (1.05 g, 7.0 mmol) and iodine (14 mg, 0.01 eq) were added, and the mixture was heated to reflux for 6 hours. After confirming the reaction by TLC, the mixture was slowly cooled. All solvents were then removed by vacuum distillation. The mixture was then diluted with 60 mL of ethyl acetate and washed with 1N-NaOH solution and brine. After drying with anhydrous Na₂SO₄, the mixture was filtered under reduced pressure. The filtrate was then distilled under reduced pressure to remove all residues, yielding a pale yellow solid.

[0229] (1.33 g)

[0230] Yield: 69.2%

[0231] 1H 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)

[0232] [Step 2] Synthesis of 6-(4-amino-3-nitrophenyl)-3-(2-(4-hydroxypiperidin-1-yl)ethyl)quinazolin-4(3H)-one (Formula 2-4)

[0233] Add 6-bromo-3-(2-(4-hydroxypiperidin-1-yl)ethyl)quinazolin-4(3H)-one (1.33 g, 3.78 mmol), 2-nitro-4-(4,4,5,5-tetramethyl-1,3,2-diaxopentenylborane-2-yl)aniline (1.0 g, 3.78 mmol), K2CO3 (1.56 g, 11.3 mmol), Pd(dppf)Cl2 (0.13 g, 0.18 mmol), and 25 ml of wet dioxane, and stir under a nitrogen atmosphere.

[0234] [1,1'-bis(diphenylphosphine)ferrocene]palladium(II) dichloride (0.13 g, 0.05 eq) was then added, and the mixture was heated under reflux for 5 hours. After confirming the reaction by TLC, the mixture was slowly cooled. After cooling, the mixture was filtered through diatomaceous earth and the filtrate was distilled under reduced pressure. Ethyl acetate was then added, stirred, and filtered to give a reddish-brown solid.

[0235] (1.2 g)

[0236] Yield: 77.4%

[0237] 1H 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)

[0238] [Step 3] Synthesis of 6-(3,4-Diaminophenyl)-3-(2-(4-hydroxypiperidin-1-yl)ethyl)quinazolin-4(3H)-one (Formula 3-4)

[0239] 1.55 g (3.7 mmol) of 6-(4-amino-3-nitrophenyl)-3-(2-(4-hydroxypiperidin-1-yl)ethyl)quinazolin-4(3H)-one was dissolved in 80 mL of methanol, and 0.77 g of 10%-Pd / C was added with stirring. Then, hydrazine hydrate (1.89 g, 37.8 mmol) was added, and the mixture was heated to reflux for 7 hours. After confirming the reaction by TLC, the mixture was cooled to room temperature. The solution was filtered through celite and then distilled under reduced pressure. After dissolving in 20 mL of methylene chloride, the solution was dried over anhydrous magnesium sulfate. The solution was filtered and distilled under reduced pressure to give a brown solid.

[0240] (1.16 g)

[0241] Yield 80.5%

[0242] 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)

[0243] [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)

[0244] 6-(3,4-diaminophenyl)-3-(2-(4-hydroxypiperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.15 g, 0.39 mmol) and 3-chloro-4-fluorobenzaldehyde (63 mg, 0.39 mmol) were dissolved in 10 mL of DMF, and sodium metabisulfite (Na₂S₂O₅, 100 mg) was added. The mixture was stirred at 95 °C for 3 hours. After confirming the reaction by TLC, the mixture was cooled, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO₄, filtered, and concentrated under reduced pressure. The resulting solid was purified by column chromatography to obtain a yellow solid.

[0245] (33 mg)

[0246] Yield 16.1%

[0247] 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, -C H -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)

[0248] 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)

[0249] 6-(3,4-diaminophenyl)-3-(2-(4-hydroxypiperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.15 g, 0.39 mmol) and 4-chlorobenzaldehyde (56 mg, 0.39 mmol) were dissolved in 10 mL of DMF, and sodium metabisulfite (Na₂S₂O₅, 100 mg) was added. The mixture was stirred at 95 °C for 3 hours. After confirming the reaction by TLC, the mixture was cooled, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO₄, filtered, and concentrated under reduced pressure. The resulting product was purified by column chromatography to give a pale yellow solid (0.48 mg).

[0250] Yield 24.3%

[0251] 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)

[0252] 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)

[0253] 6-(3,4-diaminophenyl)-3-(2-(4-hydroxypiperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.15 g, 0.39 mmol) and 4-methylbenzaldehyde (47 mg, 0.39 mmol) were dissolved in 10 mL of DMF, and sodium metabisulfite (Na₂S₂O₅, 100 mg) was added. The mixture was stirred at 95 °C for 3 hours. After confirming the reaction by TLC, the mixture was cooled, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO₄, filtered, and concentrated under reduced pressure. The resulting product was purified by column chromatography to give a pale yellow solid (20 mg).

[0254] Yield 10.5%

[0255] 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)

[0256] 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)

[0257] 6-(3,4-diaminophenyl)-3-(2-(4-hydroxypiperidin-1-yl)ethyl)quinazolin-4(3H)-one (0.15 g, 0.39 mmol) and 4-methoxybenzaldehyde (54 mg, 0.39 mmol) 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, and then 30 mL of ethyl acetate was added, followed by washing three times with 10 mL of pure water. The solution was then dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. The resulting product was purified by column chromatography to obtain a pale yellow solid.

[0258] (21 mg)

[0259] Yield 10.7%

[0260] 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)

[0261] Example 2. Measurement of anticancer activity of quinazoline-benzimidazole derivatives and confirmation of mechanism of action

[0262] 2.1 Cell viability screening of quinazoline-benzimidazole derivatives

[0263] This experiment used MDA-MB-231, a human triple-negative breast cancer (TNBC) cell line (cell seeding rate: 0.8 (M231) x 10⁻⁶). 4 Cells / well (confluence ≥ 25%) and JIMT-1 as a HER2-positive breast cancer (HER2+BC) cell line (cell seeding density: 0.8 (JIMT) x 10⁻⁶). 4 Cells / well (confluence ≥ 25%).

[0264] The cell lines were cultured at 5% CO2 and 37°C in Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum (FBS), streptomycin-penicillin (100 U / mL), and fungizone (0.625 μg / mL).

[0265] The human breast cancer cell lines MDA-MB-231 and JIMT-1 were treated with different concentrations (0, 1, and 5 μM) of quinazoline-benzimidazole derivatives of formulas 1-1 to 1-31 for 72 hours, and cell viability was then determined using the MTS assay. The MTS assay method was as follows: cells were seeded in 96-well plates and adhered for 24 hours, then treated with the quinazoline-benzimidazole derivatives for 72 hours, and then developed with MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazole) for 4 hours. The absorbance was then measured at 490 nm using a Spectramax Plus 384 microplate reader.

[0266] The measurement results are as follows Figures 1 to 3 As shown in the results, it can be confirmed that the inhibitory effects of the quinazoline-benzimidazole derivatives of formulas 1-1 to 1-31 on the cell survival of human breast cancer cell lines MDA-MB-231 and JIMT-1 are mostly concentration-dependent.

[0267] 2.2 Confirmation of the apoptosis-inducing effect of quinazoline-benzimidazole derivatives

[0268] The following experiments were conducted using compounds of formulas 1-15, 1-16, 1-19, and 1-22, which exhibited excellent activity in Examples 2-1. Trastuzumab-sensitive HER2-positive breast cancer cell lines SKBR3 and trastuzumab-resistant JIMT-1 were treated with the above compounds at different concentrations (0, 0.1, 0.5, 1, 2, and 5 μM) for 72 hours. A concentration-dependent inhibition of cell viability (MTS assay) and the IC50 values ​​for inhibiting cell viability were observed, as shown in the following figures. Figure 4 As shown in A. Additionally, 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, cell morphological changes were observed using an optical microscope, and the results are shown in Figure A. Figure 4 As shown in B. Treatment with quinazoline-benzimidazole derivatives resulted in breast cancer cell lines exhibiting typical apoptotic morphology.

[0269] Next, after treating SKBR3 and JIMT-1 cell lines with compounds of formula 1-16 and 1-19 at a concentration of 5 μM for 48 hours, the degree of apoptosis was determined by flow cytometry using Annexin V / PI staining. The results are as follows: Figure 5As shown in Figure A, the quinazoline-benzimidazole derivative effectively induced both early and late apoptosis in both cell lines. After the same treatment as above, Western blotting confirmed the expression of apoptosis-related proteins and the degradation of tubulin, as shown in Figure A. Figure 5 As shown in B. This confirms that the compounds of the present invention can achieve apoptosis by inhibiting tubulin polymerization.

[0270] 2.3 Confirmation of the mechanism of action of quinazoline-benzimidazole derivatives

[0271] As described above, the compounds in the quinazoline-benzimidazole derivatives of the present invention are designed such that their quinazoline moiety can strongly bind to the tyrosine kinase of human epidermal growth factor receptor 2 (HER2), while the benzimidazole moiety binds to the tubulin polymers to inhibit the polymerization reaction.

[0272] To verify this, the following experiments were performed. Trastuzumab-resistant HER2-positive breast cancer cells (JIMT-1) were treated with compounds of formulas 1-16 and 1-19 at a concentration of 5 μM for 48 hours. After staining with propidium iodide (PI), cell cycle analysis based on DNA content was performed by flow cytometry. The results are as follows: Figure 6 As shown in A, it can be confirmed that treatment with quinazoline-benzimidazole derivatives can induce G2 / M phase cell cycle arrest in JIMT-1 cells.

[0273] In addition, after treating JIMT-1 cell lines with 5 μM compounds 1-16 and 1-19 for 24 hours, the expression of phosphorylated histone H3, a marker of mitosis, was confirmed by immunocytochemistry. Figure 6 As shown in B, it can be confirmed that treatment with the quinazoline-benzimidazole derivative increases the expression of aberrantly phosphorylated histone H3 (phospho-Histone H3). Therefore, it can be confirmed that the quinazoline-benzimidazole derivative of the present invention can induce cell cycle arrest by inhibiting tubulin polymerization.

[0274] Then, after treating JIMT-1 cell lines with compounds 1-16 and 1-19 at a concentration of 5 μM for 24, 48, and 72 hours, the protein expression and phosphorylation levels of HER2 and HER family members (HER3, EGFR), as well as downstream signaling pathways AKT and MEK, were determined by Western blotting. The results are as follows: Figure 7 As shown, treatment with the quinazoline-benzimidazole derivative of the present invention reduced the expression and phosphorylation levels of major proteins in the time-dependent HER2 signaling pathway, particularly effectively inhibiting the activities of HER2, p95HER2, HER3, EGFR, AKT, and MEK, which are the main causes of trastuzumab resistance. Therefore, this confirms that the quinazoline-benzimidazole derivative of the present invention has the inhibitory effect on HER2 tyrosine kinases as described above, meeting the design objectives.

[0275] Example 3. Confirmation of anticancer activity of quinazoline-benzimidazole derivatives against different cancers

[0276] Different concentrations (0, 0.1, 0.5, 1, 5, 10 μM) of quinazoline-benzimidazole derivative compounds 1-16 and 1-19 were used to treat lung cancer cell line H1299, colorectal cancer cell line HCT116, ovarian cancer cell line SKOV3, prostate cancer cell line Du145, liver cancer cell line HepG2, and hematologic malignancy cell line HL-60 for 72 hours. Subsequently, a concentration-dependent inhibitory effect on cell viability (MTS assay) and the IC50 value for inhibiting cell viability were observed. The results are as follows: Figure 8 As shown in the figure, other cancer cell lines treated with the quinazoline-benzimidazole derivative of the present invention all showed effective cell viability inhibition.

[0277] The embodiments have been described above with reference to the accompanying drawings. Those skilled in the art should understand that various modifications and equivalent embodiments can be implemented based on these drawings. Suitable results can also be obtained if the described techniques are performed in a different order, and / or if the constituent elements of the described systems, architectures, devices, or circuits are combined in different ways, and / or replaced or substituted by other components or their equivalents.

[0278] Therefore, other embodiments, examples, and equivalents of the claims should be interpreted as being included within the scope of protection of the claims.

Claims

1. A quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof represented by the following [Formula 1]: [Formula 1] ； In Equation 1, R 1 It is -(CH2) n -A, A is C3-C, whether substituted or unsubstituted. 20 cycloalkyl or C3-C 20 Any of the heterocyclic alkyl groups, If the cycloalkyl or heterocycloalkyl group is substituted, it is by any one or more substitutions selected from the group consisting of C1-C6 alkyl, hydroxyl, amino, methoxy, and halogen groups. R 2 Choose C3-C, whether substituted or unsubstituted. 20 Aryl, substituted or unsubstituted C3-C 20 heteroaryl, substituted or unsubstituted C3-C 20 cycloalkyl, substituted or unsubstituted C3-C 20 Any one of the group consisting of heterocyclic alkyl groups and methyl carbamates. If the aryl, cycloalkyl, or heterocycloalkyl group is substituted, it is by any one or more substitutions selected from the group consisting of C1-C6 alkyl, halogen, hydroxyl, and alkoxy groups. n is an integer from 0 to 6.

2. The quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that, In the above [Equation 1], A is C3-C, whether substituted or unsubstituted. 20 Heterocyclic alkyl groups If the heterocyclic alkyl group is substituted, it is substituted by one or more of the methyl or hydroxyl groups. R 2 It is selected from any one of the group consisting of substituted or unsubstituted phenyl, substituted or unsubstituted thiophene, substituted or unsubstituted pyridyl, substituted or unsubstituted cyclopentyl, substituted or unsubstituted cyclohexyl and -NH-(C=O)-OCH3. If the phenyl, thienyl, pyridyl, cyclopentyl, and cyclohexyl groups are substituted, it is by any one or more substitutions selected from the group consisting of methyl, methoxy, halogen, and hydroxyl groups. n is 2.

3. The quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof according to claim 2, characterized in that, In the above [Equation 1], A's C3-C 20 Heterocyclic alkyl groups are either morpholine or piperidine.

4. The quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that, The quinazoline-benzimidazole derivative represented by [Formula 1] is selected from any one of the following groups of compounds: [Equation 1-1] ； [Equation 1-2] ； [Equation 1-3] ； [Equations 1-4] ； [Equations 1-5] ； [Equations 1-6] ； [Equations 1-7] ； [Equations 1-8] ； [Equations 1-9] ； [Equation 1-10] ； [Equation 1-11] ； [Equation 1-12] ； [Equation 1-13] ； [Equation 1-14] ； [Equation 1-15] ； [Equation 1-16] ； [Equation 1-17] ； [Equation 1-18] ； [Equation 1-19] ； [Equation 1-20] ； [Equation 1-21] ； [Equation 1-22] ； [Equation 1-23] ； [Equation 1-24] ； [Equation 1-25] ； [Equation 1-26] ； [Equation 1-27] ； [Equation 1-28] ； [Equation 1-29] ； [Equation 1-30] ； [Equation 1-31] 。 5. The quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that, The quinazoline-benzimidazole derivative represented by [Formula 1] is selected from any one of the following groups of compounds: [Equations 1-9] ； [Equation 1-10] ； [Equation 1-11] ； [Equation 1-13] ； [Equation 1-15] ； [Equation 1-16] ； [Equation 1-17] ； [Equation 1-20] ； [Equation 1-22] 。 6. The quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that, The quinazoline-benzimidazole derivative has the effect of inhibiting microtubule polymerization and inhibiting human epidermal growth factor receptor 2 (HER2) tyrosine kinase.

7. The quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that, The pharmaceutically acceptable salt of the quinazoline-benzimidazole derivative is selected from any one or more of the group consisting of hydrochloride, bromate, sulfate, phosphate, nitrate, citrate, acetate, lactate, tartrate, maleate, gluconate, succinate, formate, trifluoroacetate, oxalate, fumarate, glutarate, adipate, methanesulfonate, benzenesulfonate, p-toluenesulfonate, camphorsulfonate, sodium salt, potassium salt, lithium salt, calcium salt, and magnesium salt.

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

9. The pharmaceutical composition for preventing or treating cancer according to claim 8, characterized in that, The pharmaceutical composition induces apoptosis by causing cell cycle arrest in cancer cells.

10. The pharmaceutical composition for preventing or treating cancer according to claim 8, characterized in that, The pharmaceutical composition inhibits the activity of any one or more of HER2, p95HER2, HER3, EGFR, AKT, and MEK.

11. The pharmaceutical composition for preventing or treating cancer according to claim 8, characterized in that, The cancer is selected from any one or more of the following groups: skin cancer, breast cancer, uterine cancer, esophageal cancer, stomach cancer, brain tumor, colon cancer, rectal cancer, colorectal cancer, lung cancer, ovarian cancer, cervical cancer, endometrial cancer, vulvar cancer, kidney cancer, hematologic malignancies, pancreatic cancer, prostate cancer, testicular cancer, laryngeal cancer, head and neck cancer, thyroid cancer, liver cancer, bladder cancer, osteosarcoma, lymphoma, hematologic malignancies, thymic cancer, urethral cancer, and bronchial cancer.

12. The pharmaceutical composition for preventing or treating cancer according to claim 8, characterized in that, The cancer in question is either HER2-positive breast cancer or triple-negative breast cancer.

13. A method for preparing a quinazoline-benzimidazole derivative or a pharmaceutically acceptable salt thereof represented by the following [Formula 1], wherein the derivative is prepared by the following steps: 1) By making R 1 The steps for preparing the compound represented by the following [Formula 4] by reacting the compound represented by -NH2 with 5-bromo-o-aminobenzoic acid and triethyl orthoformate; 2) The step of preparing the compound represented by [Formula 2] by reacting the compound represented by [Formula 4] with either 2-nitro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaneborane-2-yl)aniline or 4-amino-3-nitrophenylboronic acid pinacol ester; 3) The steps for preparing the compound represented by [Formula 3] from the compound represented by [Formula 2]; and 4) By reacting the compound represented by [Equation 3] with... The compound represented by R or by R 2 The steps for preparing the compound represented by [Formula 1] by reacting any of the compounds represented by -CHO; [Formula 1] ； [Equation 2] ； [Formula 3] ； [Formula 4] ； In equations 1 to 4, R 1 It is -(CH2) n -A, A is C3-C, whether substituted or unsubstituted. 20 cycloalkyl or C3-C 20 Any of the heterocyclic alkyl groups, If the cycloalkyl or heterocycloalkyl group is substituted, it is by any one or more substitutions selected from the group consisting of C1-C6 alkyl, hydroxyl, amino, methoxy, and halogen groups. R 2 Choose C3-C, whether substituted or unsubstituted. 20 Aryl, substituted or unsubstituted C3-C 20 heteroaryl, substituted or unsubstituted C3-C 20 cycloalkyl, substituted or unsubstituted C3-C 20 Any one of the group consisting of heterocyclic alkyl groups and methyl carbamates. If the aryl, cycloalkyl, or heterocycloalkyl group is substituted, it is substituted by one or more of the group consisting of C1-C6 alkyl, halogen, hydroxyl, and alkoxy groups, where n is an integer from 0 to 6.

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