Thiobenzimidazole derivatives or pharmaceutically acceptable salts thereof and uses thereof

Abstract

The present invention relates to a thiobenzimidazole derivative or a pharmaceutically acceptable salt thereof, and a composition for preventing or treating cancer containing the derivative as an active ingredient. The thiobenzimidazole derivative of the present invention is activated in cancer cells to inhibit tubulin polymerization, and when administered to an individual, can block the cancer cell cycle and induce cell apoptosis to produce cytotoxicity. The thiobenzimidazole derivative can be used to prevent or treat cancer, preferably for preventing or treating triple-negative breast cancer.

Description

Technical Field

[0001] The present invention relates to thiobenzimidazole derivatives or pharmaceutically acceptable salts thereof and uses thereof. Background Art

[0002] Triple-negative breast cancer (TNBC, ER-, PR-, HER2-) patients account for 10% to 15% of all breast cancer patients. Due to the lack of hormone receptors (ER (estrogen receptor), PR (progesterone receptor)) and HER2 protein, they cannot benefit from hormone therapy or HER2 targeted therapy. The current standard treatment for triple-negative breast cancer relies entirely on general cytotoxic anticancer drugs (taxanes or anthracyclines). Due to the lack of clear targeted therapeutic drugs, the treatment strategies for other subtypes are very simple compared to breast cancer. What is more serious is that most patients will relapse within 2 to 3 years after surgery or chemotherapy, and it is also easy to induce metastasis to other organs such as the lungs, liver, brain, and bones, reducing the patient's survival rate.

[0003] The 5-year overall survival rate for stage III patients is less than 55%, and for patients with advanced-stage disease who have already developed metastatic disease (advanced-stage disease), the 5-year survival rate is less than 30%, which is a very low level. Most of these patients develop very serious disease and die within a few years.

[0004] Microtubules, a major component of the cytoskeleton, are composed of tubulin heteropolymers based on α- and β-subunits. Microtubules perform numerous cellular functions, such as intracellular transport, maintaining polarity, transmitting intracellular signals, cell migration, and proliferation. During mitosis, spindle fibers form, aligning chromosomes at the cell center and then separating toward the poles. Failure of the spindle to function properly can inhibit cell division and lead to apoptosis, thus attracting attention as a target for anticancer drugs.

[0005] Drugs targeting microtubules are generally divided into drugs that stabilize microtubules and drugs that destabilize microtubules. First, microtubule stabilizers include taxanes, paclitaxel (Taxol) and docetaxel (decetaxel), which can prevent microtubule depolymerization and promote polymerization. Most microtubule stabilizing substances bind to taxane binding sites or overlapping sites of β-tubulin. Secondly, microtubule destabilizers include colchicine and vinca alkaloids, which bind to colchicine binding sites or vinca binding sites. Compared with drugs that affect microtubule polymers, drugs that target microtubules themselves can work at lower drug concentrations and ultimately inhibit cell mitosis. Therefore, there is a need to develop potential microtubule polymerization inhibitors as anticancer agents.

[0006] Meanwhile, commercially available anthelmintics contain tubulin polymerization inhibitors, flubendazole and albendazole. These inhibitors have been shown to induce cell death through cell cycle arrest and have apoptotic effects on slow-growing cancer cells, attracting attention as cancer treatments. However, their low solubility in water makes them difficult to absorb, and excessive consumption can lead to side effects such as elevated liver enzymes, necessitating the development of derivatives with novel structures.

[0007] Therefore, the present inventors confirmed that novel thiobenzimidazole derivatives or pharmaceutically acceptable salts thereof can act as tubulin polymerization inhibitors to achieve cell cycle arrest of cancer cells, thereby inducing apoptosis, thereby completing the present invention. Summary of the Invention

[0008] Technical problems to be solved

[0009] The object of the present invention is to provide a thiobenzimidazole derivative or a pharmaceutically acceptable salt thereof.

[0010] The object of the present invention is to provide a method for preparing a thiobenzimidazole derivative or a pharmaceutically acceptable salt thereof.

[0011] Another object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer, comprising a thiobenzimidazole derivative or a pharmaceutically acceptable salt thereof as an active ingredient.

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

[0013] Technical methods to solve problems

[0014] To solve the above problems, the present invention provides a thiobenzimidazole derivative or a pharmaceutically acceptable salt thereof as shown in Chemical Formula 1.

[0015] [Chemical Formula 1]

[0016]

[0017] In the chemical formula 1,

[0018] R1 is -S-R2 or -SS-R2,

[0019] R2 is any one selected from the group consisting of substituted or unsubstituted cycloalkyl, heterocycloalkyl, aryl, heteroaryl and alkylaryl, preferably a substituted or unsubstituted C3 to C 20 Aryl, substituted or unsubstituted C3 to C 20 Heteroaryl, substituted or unsubstituted C3 to C 20 Alkylaryl.

[0020] In one embodiment of the present invention, in the chemical formula 1, the substituted aryl, heteroaryl or alkylaryl may be substituted by at least one selected from the group consisting of substituted or unsubstituted alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, halogen, hydroxyl, alkoxy and amino, preferably substituted or unsubstituted C3 to C 12 Heterocycloalkyl, substituted or unsubstituted C3 to C 12 substituted with heteroaryl and halogen.

[0021] In one embodiment of the present invention, the thiobenzimidazole derivative represented by Chemical Formula 1 is selected from any one of the group consisting of the following compounds:

[0022] [Chemical Formula 1-1]

[0023]

[0024] [Chemical formula 1-2]

[0025]

[0026] [Chemical formula 1-3]

[0027]

[0028] [Chemical formula 1-4]

[0029]

[0030] [Chemical Formula 1-5]

[0031]

[0032] [Chemical formula 1-6]

[0033]

[0034] [Chemical Formula 1-7]

[0035]

[0036] [Chemical Formula 1-8]

[0037]

[0038] [Chemical Formula 1-9]

[0039]

[0040] [Chemical formula 1-10]

[0041]

[0042] [Chemical Formula 1-11]

[0043]

[0044] [Chemical formula 1-12]

[0045]

[0046] [Chemical Formula 1-13]

[0047]

[0048] [Chemical Formula 1-14]

[0049]

[0050] [Chemical Formula 1-15]

[0051]

[0052] [Chemical Formula 1-16]

[0053]

[0054] [Chemical Formula 1-17]

[0055]

[0056] [Chemical Formula 1-18]

[0057]

[0058] [Chemical Formula 1-19]

[0059]

[0060] [Chemical formula 1-20]

[0061]

[0062] [Chemical Formula 1-21]

[0063] as well as

[0064] [Chemical formula 1-22]

[0065]

[0066] As another embodiment of the present invention, the thiobenzimidazole derivative can inhibit tubulin polymerization.

[0067] As another embodiment of the present invention, the pharmaceutically acceptable salt of the thiobenzimidazole derivative can be selected from any one 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.

[0068] In addition, the present invention provides a method for preparing the hydrochloride of thiobenzimidazole, comprising the following steps:

[0069] (1) preparing a suspension of a thiobenzimidazole derivative represented by Chemical Formula 1;

[0070] [Chemical Formula 1]

[0071]

[0072] (2) injecting hydrogen chloride (HCl) into the suspension and evaporating it under reduced pressure;

[0073] (3) adding isopropyl alcohol to the product of step (2), heating, and then cooling; and

[0074] (4) Add isopropyl ether to the product of step (3), stir, and then filter.

[0075] In addition, the present invention provides a pharmaceutical composition for preventing or treating cancer, which comprises the thiobenzimidazole derivative or a pharmaceutically acceptable salt thereof as an effective ingredient.

[0076] In addition, the present invention provides a method for preventing or treating cancer, comprising the step of administering the thiobenzimidazole derivative or a pharmaceutically acceptable salt thereof to a subject.

[0077] The present invention provides a use of the thiobenzimidazole derivative or a pharmaceutically acceptable salt thereof for preparing a drug for preventing or treating cancer.

[0078] Furthermore, the present invention provides a method for diagnosing cancer, comprising the step of administering the thiobenzimidazole derivative or a pharmaceutically acceptable salt thereof to a subject.

[0079] The present invention provides a use of the thiobenzimidazole derivative or a pharmaceutically acceptable salt thereof for preparing cancer diagnostic drugs.

[0080] In one embodiment of the present invention, the pharmaceutical composition induces cell apoptosis by arresting the cancer cell cycle.

[0081] In another embodiment of the present invention, the cancer can be selected from at least any one 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, blood cancer, pancreatic cancer, prostate cancer, testicular cancer, laryngeal cancer, head and neck cancer, thyroid cancer, liver cancer, bladder cancer, osteosarcoma, lymphoma, blood cancer, thymus cancer, urethral cancer and bronchial cancer, preferably breast cancer, more preferably triple-negative breast cancer.

[0082] Effects of the Invention

[0083] The present invention relates to a thiobenzimidazole derivative or a pharmaceutically acceptable salt thereof and a composition for preventing or treating cancer containing the derivative as an active ingredient. The thiobenzimidazole derivative of the present invention is activated in cancer cells to inhibit tubulin polymerization and, when administered to an individual, can block the cancer cell cycle and induce cell apoptosis, thereby producing cytotoxicity. The thiobenzimidazole derivative can be used to prevent or treat cancer, preferably for preventing or treating triple-negative breast cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0084] Figure 1The results show the cell survival rates of the triple-negative breast cancer (TNBC) cell line MDA-MB-231 after treatment with the thiobenzimidazole derivatives of Chemical Formulas 1-1, 1-2, 1-3, and 1-5.

[0085] Figure 2 The results show the cell survival rates of the HER2-positive breast cancer (HER2+BC) cell line JIMT-1 after treatment with the thiobenzimidazole derivatives of Chemical Formulas 1-1, 1-2, 1-3, and 1-5.

[0086] Figure 3 The results show the cell survival rates of the triple-negative breast cancer (TNBC) cell line MDA-MB-231 after treatment with the thiobenzimidazole derivatives of Chemical Formulas 1-6, 1-7, 1-8, 1-10, 1-11, 1-12, 1-13, and 1-14.

[0087] Figure 4 The results of cell survival rates of HER2-positive breast cancer cells JIMT-1 after treatment with the thiobenzimidazole derivatives of Chemical Formulas 1-6, 1-7, 1-8, 1-10, 1-11, 1-12, 1-13, and 1-14 are shown.

[0088] Figure 5 The cell survival results of the triple-negative breast cancer (TNBC) cell line MDA-MB-231 after being treated with the thiobenzimidazole derivatives of Chemical Formulas 1-15, 1-16, 1-17, 1-19, 1-20, 1-21 and 1-22.

[0089] Figure 6 The results show the cell survival rates of HER2-positive breast cancer cells JIMT-1 after treatment with the thiobenzimidazole derivatives of Chemical Formulas 1-15, 1-16, 1-17, 1-19, 1-20, 1-21, and 1-22.

[0090] Figure 7 The cell survival results of HER2-positive breast cancer cell lines SKBR3, BT474, and JIMT-1 after being treated with the thiobenzimidazole derivatives of Chemical Formulas 1-3 are shown.

[0091] Figure 8 Results showing confirmation that the thiobenzimidazole derivatives of Chemical Formulas 1-3 induce cancer cell death (Sub-G1 accumulation) and G2 / M phase cell cycle arrest.

[0092] Figure 9Results showing the early and late apoptosis of cancer cells confirmed by the thiobenzimidazole derivatives of Chemical Formulas 1-3.

[0093] Figure 10 The cell survival results of HER2-positive breast cancer cell lines BT474 and JIMT-1 after being treated with the thiobenzimidazole derivatives of Chemical Formula 1-3 and their hydrochlorides (Compound 1-23) are shown.

[0094] Figure 11 The cell survival results of triple-negative breast cancer (TNBC) cell lines MDA-MB-231, BT549, and 4T1 after being treated with the hydrochloride salt of the thiobenzimidazole derivative of Chemical Formula 1-3 (Compound 1-23) are shown.

[0095] Figure 12 Western blot results are shown for triple-negative breast cancer (TNBC) cell line MDA-MB-231 and HER2-positive breast cancer cell line JIMT-1 after treatment with the hydrochloride salt of the thiobenzimidazole derivative of Chemical Formula 1-3 (Compound 1-23).

[0096] Figure 13 The results show the cell survival rates of blood cancer cell line HL-60, colorectal cancer cell line HCT116, and non-small cell lung cancer cell lines H1299 and A549 after being treated with the hydrochloride of the thiobenzimidazole derivative of Chemical Formula 1-3 (Compound 1-23).

[0097] Figure 14 The results show the cell survival rates of ovarian cancer cell line SKOV3, prostate cancer cell line Du145, and liver cancer cell line HepG2 after being treated with the hydrochloride of the thiobenzimidazole derivative of Chemical Formula 1-3 (Compound 1-23). DETAILED DESCRIPTION

[0098] The present invention is completed by conducting in-depth research on thiobenzimidazole derivatives or pharmaceutically acceptable salts thereof and confirming the anticancer activity of the derivatives.

[0099] More specifically, it was confirmed that the thiobenzimidazole derivative or a pharmaceutically acceptable salt thereof has the following characteristics: it is activated in cancer cells to inhibit tubulin polymerization, arrest cancer cell cycle, and induce cell apoptosis, thereby exhibiting cytotoxicity.

[0100] From the above results, it can be seen that the present invention can provide a thiobenzimidazole derivative represented by the following [Chemical Formula 1] or a pharmaceutically acceptable salt thereof.

[0101] [Chemical Formula 1]

[0102]

[0103] In the chemical formula,

[0104] R1 is -S-R2 or -SS-R2,

[0105] R2 may be optionally substituted or unsubstituted C3 to C 20 Aryl, substituted or unsubstituted C3 to C 20 Heteroaryl, and substituted or unsubstituted C3 to C 20 The group consisting of alkylaryl groups.

[0106] In the present invention, the term "substitution" refers to a reaction in which one atom or atomic group in a compound molecule is replaced by another atom or atomic group.

[0107] In the present invention, the term "chain" refers to a molecule having a chain structure, which is a chemical structure in which carbon atoms are connected in a chain, and can be a straight chain or a branched chain.

[0108] In the present invention, the term "cyclic" refers to a ring structure formed by connecting two ends of an organic compound skeleton.

[0109] In the present invention, the term "chain or cyclic alkyl group" refers to a monovalent linear or branched or cyclic saturated hydrocarbon residue having 1 to 20 carbon atoms and consisting solely of carbon and hydrogen atoms. Non-limiting examples of such alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, 2-butyl, 3-butyl, pentyl, n-hexyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0110] In the present invention, the term "heterocycloalkyl" generally refers to a saturated or unsaturated (but not aromatic) cyclic hydrocarbon (cyclohydrocarbon), which may be unsubstituted, monosubstituted or polysubstituted, and has at least one heteroatom selected from N, O or S in its structure.

[0111] In the present invention, the term "aryl" refers to an unsaturated aromatic ring compound having 3 to 12 carbon atoms and having a single ring (e.g., phenyl) or multiple condensed rings (e.g., naphthyl). Non-limiting examples of such aryl groups include phenyl, naphthyl, etc.

[0112] In the present invention, the term "heteroaryl" refers to a monocyclic or condensed ring having at least one heteroatom of N, O or S among the atoms constituting the ring. Non-limiting examples of such heteroaryl groups include pyridyl, pyrimidinyl, pyrazinyl, oxazolyl, furyl and the like.

[0113] In the present invention, the term "alkoxy" refers to an alkyl group bonded to oxygen (-OR). Non-limiting examples of such alkoxy groups include methoxy, ethoxy, propoxy, butoxy, and the like.

[0114] In the present invention, "halogen" may be fluorine (F), chlorine (Cl), bromine (Br), or iodine (I).

[0115] In the present invention, more specifically, the R2 can be

[0116]

[0117] In a preferred embodiment of the present invention, the compound represented by Chemical Formula 1 is preferably at least one selected from the group consisting of the following compounds.

[0118] [Chemical Formula 1-1]

[0119]

[0120] [Chemical formula 1-2]

[0121]

[0122] [Chemical formula 1-3]

[0123]

[0124] [Chemical formula 1-4]

[0125]

[0126] [Chemical Formula 1-5]

[0127]

[0128] [Chemical formula 1-6]

[0129]

[0130] [Chemical Formula 1-7]

[0131]

[0132] [Chemical Formula 1-8]

[0133]

[0134] [Chemical Formula 1-9]

[0135]

[0136] [Chemical formula 1-10]

[0137]

[0138] [Chemical Formula 1-11]

[0139]

[0140] [Chemical formula 1-12]

[0141]

[0142] [Chemical Formula 1-13]

[0143]

[0144] [Chemical Formula 1-14]

[0145]

[0146] [Chemical Formula 1-15]

[0147]

[0148] [Chemical Formula 1-16]

[0149]

[0150] [Chemical Formula 1-17]

[0151]

[0152] [Chemical Formula 1-18]

[0153]

[0154] [Chemical Formula 1-19]

[0155]

[0156] [Chemical formula 1-20]

[0157]

[0158] [Chemical Formula 1-21]

[0159] And [Chemical Formula 1-22]

[0160]

[0161] The thiobenzimidazole derivative or its pharmaceutically acceptable salt of the present invention is specifically activated in cancer cells, can inhibit tubulin polymerization and induce cell death, and can be used in a pharmaceutical composition for preventing or treating cancer with the thiobenzimidazole derivative or its pharmaceutically acceptable salt as an active ingredient.

[0162] In the present invention, the term "pharmaceutically acceptable salt" refers to a dosage form of a compound that does not cause severe irritation to the organism to which the compound is administered and does not impair the biological activity and physical properties of the compound. Pharmaceutically acceptable salts can be prepared by reacting the compound of the present invention with an inorganic acid such as hydrochloric acid, bromic acid, sulfuric acid, nitric acid, phosphoric acid; a sulfonic acid such as methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid; an organic carbonic acid 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, salicylic acid, etc. In addition, the compound of the present invention can be reacted with a base to form an alkali metal salt such as ammonium salt, sodium salt, potassium salt; an alkaline earth metal salt such as calcium salt, magnesium salt; a salt of an organic base such as dicyclohexylamine, N-methyl-D-glucamine, tris(hydroxymethyl)methylamine; and an amino acid salt such as arginine and lysine.

[0163] In addition, the thiobenzimidazole derivative or a pharmaceutically acceptable salt thereof includes not only pharmaceutically acceptable salts but also all salts, hydrates and solvates that can be prepared by conventional methods.

[0164] Furthermore, the present invention may provide a method for preventing, treating and / or diagnosing cancer, comprising the step of administering the thiobenzimidazole derivative or a pharmaceutically acceptable salt thereof to a subject.

[0165] Herein, the term "prevention" refers to all actions that inhibit or delay the occurrence, spread or recurrence of cancer by administering the composition of the present invention; "treatment" refers to all actions that improve or benefit cancer symptoms by administering the composition of the present invention.

[0166] As used herein, the term "pharmaceutical composition" is intended for the prevention or treatment of a disease and can be formulated into various forms according to conventional methods. For example, it can be formulated into oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, and syrups, as well as external use preparations, suppositories, and sterile injections.

[0167] In the present invention, "comprising as an active ingredient" means that the amount of the ingredient is sufficient or necessary to achieve the desired biological effect. In practical applications, the amount of the active ingredient is determined based on the intended therapeutic purpose and should not cause other toxicities. For example, the amount may vary depending 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 can empirically determine the effective amount of a single composition without the need for undue experimentation.

[0168] Furthermore, the pharmaceutical composition of the present invention may contain one or more pharmaceutically acceptable carriers in addition to the active ingredients of the above-mentioned dosage forms.

[0169] The pharmaceutically acceptable carrier can be saline, sterile water, Ringer's solution, buffered saline, glucose solution, maltodextrin solution, glycerol, ethanol and at least one mixture of these components, and can further include other common additives such as antioxidants, buffers, antibacterial agents as needed. In addition, diluents, dispersants, surfactants, adhesives and lubricants can be further added to make injection preparations such as aqueous solutions, suspensions, emulsions, pills, capsules, granules or tablets. In addition, it is preferably possible to use a suitable method in the art or use the method disclosed in Lei's Pharmaceutical Science (Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA) to configure to adapt to different diseases or components.

[0170] The composition of the present invention can be administered orally or parenterally in a pharmaceutically effective amount according to the desired method. The term "pharmaceutically effective amount" of the present invention refers to an amount that can fully treat the disease and does not cause side effects within the reasonable benefit / risk ratio of applicable drug treatment. The level of the effective amount can be determined based on many factors and other factors well known in the medical field. Factors to be considered may include the patient's health status, severity, drug activity, drug sensitivity, administration method, administration time, administration route and excretion rate, treatment time, and drugs prepared or taken at the same time.

[0171] Furthermore, the pharmaceutical composition of the present invention can be administered to a subject to prevent, treat and / or diagnose cancer, including 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, blood cancer, pancreatic cancer, prostate cancer, testicular cancer, laryngeal cancer, head and neck cancer, thyroid cancer, liver cancer, bladder cancer, osteosarcoma, lymphoma, blood cancer, thymus cancer, urethra cancer, or bronchial cancer, etc., preferably a cancer with higher acidity than normal cells and whose cytotoxicity can be inhibited by microtubule polymerization inhibitors, non-limiting examples of which include breast cancer, preferably triple-negative breast cancer, etc.

[0172] In the present invention, the term "subject" may be any mammal such as livestock or humans for whom cancer prevention, treatment and / or diagnosis is required, and is not particularly limited, but is preferably a human.

[0173] The pharmaceutical composition of the present invention can be formulated into a variety of dosage forms for administration to a subject, and representative parenteral preparations are injection preparations, preferably isotonic aqueous solutions or suspensions. Injection preparations can be prepared using suitable dispersants or wetting agents and suspending agents according to techniques known in the art. For example, each component can be dissolved in physiological saline or a buffer solution to prepare an injection. In addition, dosage forms for oral administration include ingestible tablets, buccal tablets, lozenges, capsules, elixirs, suspensions, syrups, and wafers, etc., and the above-mentioned dosage forms may include diluents (e.g., lactose, glucose, sucrose, mannitol, sorbitol, cellulose, and / or glycine) and lubricants (e.g., silicon dioxide, talc, stearic acid, and its magnesium salt or calcium salt, and / or polyethylene glycol) in addition to the active ingredient. The tablets may contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth gum, methylcellulose, sodium carboxymethylcellulose, and / or polyvinyl pyrrolidone, and may further include disintegrants such as starch, agar, alginic acid or its sodium salt, absorbents, colorants, flavorings, and / or sweeteners, as appropriate. The preparations may be prepared by conventional mixing, granulation, or coating methods.

[0174] In addition, the pharmaceutical composition of the present invention may further include preservatives, hydrating agents, emulsifying agents, auxiliary agents such as salts or buffers for controlling osmotic pressure, and other substances that can be used for treatment, and can be prepared according to conventional methods.

[0175] The pharmaceutical compositions of the present invention can be administered via a variety of 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 severity of the disease. Furthermore, the compositions of the present invention can be used in conjunction with known compounds that enhance the desired effect.

[0176] The pharmaceutical composition according to the present invention can be administered to humans or animals by oral administration, or by 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 drip methods.

[0177] In the pharmaceutical composition of the present invention, the total effective amount of the thiobenzimidazole derivative or a pharmaceutically acceptable salt thereof can be administered to the patient via a single dose or multiple doses over a long period of time using a fractionated treatment protocol. The content of the active ingredient in the pharmaceutical composition of the present invention can be varied depending on the severity of the disease, but generally, based on an adult, the effective dose per administration can be 100 μg to 3,000 mg, and can be administered multiple times a day. However, the concentration of the thiobenzimidazole derivative or a pharmaceutically acceptable salt thereof depends on a variety of factors, such as the patient's age, weight, health status, sex, severity of the disease, diet, and excretion rate, and the dosage to be administered to the patient can be determined taking these factors into consideration.

[0178] In addition, as long as the pharmaceutical composition of the present invention exerts the effects of the present invention, there are no particular restrictions on its dosage form, route of administration, and method of administration. In addition to the thiobenzimidazole derivative or its pharmaceutically acceptable salt as an active ingredient, the pharmaceutical composition of the present invention may further include a known anticancer agent and can be used together with other known treatment methods.

[0179] The terms used in the examples are intended only to illustrate specific embodiments and are not intended to limit the scope. Unless otherwise specified in the context, singular expressions include the plural. In this specification, terms such as "including" or "having" are used to express the presence of features, numbers, steps, operations, constituent elements, accessories, or combinations thereof described in the specification, and do not exclude the possibility of the presence or addition of one or more other features, numbers, steps, operations, constituent elements, accessories, or combinations thereof.

[0180] Unless otherwise defined, all terms used herein, including technical or scientific terms, have the ordinary meanings understood by those skilled in the art. Commonly used terms, as defined in dictionaries, should be understood as having the meanings in the relevant technical context and, unless explicitly defined in this specification, should not be interpreted as idealizing or formalizing the meaning.

[0181] Below, the embodiments will be described in detail with reference to the accompanying drawings. It should be understood that various changes can be made to the embodiments, and the scope of the present application is not limited to the following embodiments. All changes made to the embodiments, their equivalents, and even their substitutes are within the scope of the present invention.

[0182] [Example]

[0183] Example 1. Preparation of thiobenzimidazole derivatives

[0184] All chemical reagents were commercially available products.1 H NMR spectra were recorded on a Bruker Avance III 400 MHz and a Bruker Fourier 300 MHz, with TMS used as an internal standard.

[0185] LCMS was performed using an Agilent LC / MSD 1200 series quadrupole mass spectrometer (column: ODS2000 (50×4.6 mm, 5 μm)) operating in ES (+) or (−) ionization mode, T=30° C., flow rate=1.5 mL / min; detection wavelength: 214 nm.

[0186] 1.1 Synthetic Chemical Formula 1-1

[0187] The following reaction formula 1 shows the synthesis process of the thiobenzimidazole derivative of Chemical Formula 1-1.

[0188] [Reaction formula 1]

[0189]

[0190] 1.1.1. Synthesis of 5-(4-fluorophenylthio)-2-nitrobenzeneamine

[0191] 4-Fluorothiol (1.1 g, 8.69 mmol) was dissolved in 10 mL and K2CO3 (1.24 g, 9.01 mmol) was added. 2-Nitro-5-Chloroaniline (1.5 g, 8.69 mmol) was then dissolved in 3 mL and added dropwise. The mixture was then stirred at 90°C for 3 hours. After the reaction was confirmed to be complete by TLC, it was cooled to room temperature. After pouring 50 mL of pure water, it was extracted with 100 mL of ethyl acetate. It was dried over anhydrous MgSO4 and filtered, and then distilled under reduced pressure to obtain a yellow solid, which was recrystallized from ethyl acetate / n-hexane to obtain 5-(4-fluorophenylsulfanyl)-2-nitrobenzylamine (1.87 g, yield 82%) as a yellow solid.

[0192] R f :0.37(EtOAc / n-Hexane,1:5)

[0193] 1H NMR (400MHz. CDCl3 yield: 82%, δ, ppm): 6.05 (brs, 2H, -NH2), 6.33~8.02 (m, 7H, ArH)

[0194] 1.1.2. Synthesis of 4-(4-fluorophenylthio)benzene-1,2-diamine

[0195] 5-(4-fluorophenylsulfanyl)-2-nitrobenzylamine (1.0 g, 0.378 mmol) was dissolved in 20 mL of acetic acid and then Zn (1.23 g, 18.9 mmol) was slowly added under a cooling bath. The mixture was stirred at room temperature for 10 hours and the reaction was confirmed to be complete by TLC and filtered. The filtrate was distilled under reduced pressure and extracted with 100 mL of ethyl acetate after adjusting the pH to 8 with 5M-NaOH solution. It was dried over anhydrous MgSO4, filtered, and distilled under reduced pressure to obtain a dark brown liquid, which was subjected to column chromatography to obtain 4-(4-fluorophenylsulfanyl)benzene-1,2-diamine (0.84 g, yield 95.4%) in a brown liquid state.

[0196] R f :0.06(EtOAc / n-Hexane,1:4)

[0197] 1 H NMR(400MHz.CDCl3,δ,ppm):3.41(brs,2H,-NH2),3.50(brs,2H,-NH2),6.68~7.28(m,7H,ArH)

[0198] 1.1.3. Synthesis of Methyl 5-(4-fluorophenylthio)-1H-benzo[d]imidazol-2-ylcarbamate

[0199] 4-(4-Fluorophenylsulfanyl)benzene-1,2-diamine (0.7 g, 2.98 mmol) and 1,3-bis(methoxycarbonyl)-S-methylisothiourea (1,3-Bis(methoxycarbonyl)-S-methylisothiourea, 1.6 g, 7.77 mmol) were dissolved in 10 mL of 5%-AcOH in ethanol and heated under reflux for 4 hours. After the reaction was confirmed to be complete by TLC, it was cooled to room temperature and the resulting precipitate was filtered. During filtration, the mixture was thoroughly washed with MeOH and dried in vacuo at 50 ° C to obtain methyl 5-(4-fluorophenylsulfanyl)-1H-benzo[d]imidazol-2-ylcarbamate (Chemical Formula 1-1, 1.02 g, yield 78%) as a white solid.

[0200] Melting point: 232-233℃

[0201] 1 H NMR(400MHz.CDCl3,δ,ppm):3.85(s,3H,-OCH3),6.95~7.69(m,7H,ArH),11.74(brs,2H,(-NH)2)

[0202] 1.2 Synthetic Chemical Formula 1-2

[0203] The following reaction formula 2 shows the synthesis process of the thiobenzimidazole derivative of Chemical Formula 1-2.

[0204] [Reaction formula 2]

[0205]

[0206] 1.2.1. Synthesis of tert-butyl 4-(4-((ethoxycarbonothioyl)thio)phenyl)piperazine-1-carboxylate

[0207] To a solution of tert-butyl 4-(4-aminophenyl)piperazine-1-carboxylate (4-(4-aminophenyl)piperazine-1-carboxylate, 2.77 g, 10.2 mmol) in THF / H2O (40 mL / 40 mL) was added a concentrated solution of H2SO4 (1.96 g, 20.0 mmol). NaNO2 (2.10 g, 30 mmol) dissolved in water (3.0 mL) was then slowly added at -5°C to 0°C and stirred for 2 hours. Potassium O-ethyl carbonodithioate (9.60 g, 60 mmol) was added and the mixture was stirred at room temperature for 2 hours. After completion of the reaction, the mixture was poured into water (100 mL) and extracted with EA (100 mL*3). The organic layer was dried over Na2SO4, concentrated, and purified by reverse phase column chromatography (ACN / H2O) to give 2.00 g of tert-butyl 4-(4-((ethoxythiocarbonyl)thio)phenyl)piperazine-1-carboxylate as an orange solid (yield 52%).

[0208] 1.2.2. Synthesis of tert-butyl 4-(4-((3-amino-4-nitrophenyl)thio)phenyl)piperazine-1-carboxylate

[0209] To a solution of tert-butyl 4-(4-((ethoxythiocarbonyl)thio)phenyl)piperazine-1-carboxylate (1.15 g, 3.00 mmol) in THF / MeOH / H₂O (30 mL / 50 mL / 10 mL) were added 5-chloro-2-nitroaniline (520 mg, 3.00 mmol) and NaOH (480 mg, 12 mmol), and the mixture was stirred at 65° C. for 16 hours. After completion of the reaction, the mixture was adjusted to pH 7-8 with AcOH, concentrated, and purified by reverse phase column chromatography (ACN / H₂O) to afford 600 mg of tert-butyl 4-(4-((3-amino-4-nitrophenyl)thio)phenyl)piperazine-1-carboxylate as a brown solid (yield 46%).

[0210] 1.2.3. Synthesis of tert-butyl 4-(4-((2-((methoxycarbonyl)amino)-1H-benzo[d]imidazol-6-yl)thio)phenyl)piperazine-1-carboxylate

[0211] To a solution of tert-butyl 4-(4-((3-amino-4-nitrophenyl)thio)phenyl)piperazine-1-carboxylate (500 mg, 1.16 mmol) in AcOH (30 mL) were added 1,3-bis(methoxycarbonyl)-2-methyl-2-thiopseudourea (478 mg, 2.32 mmol) and Zn powder (603 mg, 9.28 mmol), and the mixture was stirred at 75° C. for 20 hours. After the reaction was complete, the solvent was removed. The mixture was dissolved in MeOH (50 mL) and filtered. The filtrate was concentrated and purified by reverse phase column chromatography (ACN / H2O) to give 300 mg of tert-butyl 4-(4-((2-((methoxycarbonyl)amino)-1H-benzo[d]imidazol-6-yl)thio)phenyl)piperazine-1-carboxylate as a yellow solid (yield 53%).

[0212] 1.2.4. Synthesis of methyl (6-((4-(piperazin-1-yl)phenyl)thio)-1H-benzo[d]imidazol-2-yl)carbamate

[0213] A solution of tert-butyl 4-(4-((2-((methoxycarbonyl)amino)-1H-benzo[d]imidazol-6-yl)thio)phenyl)piperazine-1-carboxylate (300 mg, 0.62 mmol) in dioxane (6 mL) was added HCl / dioxane (6 mL, 6 mol / L), and the mixture was stirred at room temperature for 2 hours. After completion of the reaction, the solvent was removed and EtN was added. The mixture was concentrated and purified by perp-HPLC to give 100 mg of methyl (6-((4-(piperazin-1-yl)phenyl)thio)-1H-benzo[d]imidazol-2-yl)carbamate as an off-white solid (yield 42%).

[0214] 1 H-NMR (400MHz, DMSO-d6): δ7.33 (d, 1H, J = 8.0Hz), 7.29 (s, 1H), 7.21 (d, 1H, J = 8.8HZ), 7. 03-7.00(m,1H),6.90(d,2H,J=8.8Hz),3.74(s,3H),3.06-3.03(m,4H),2.82-2.79(m,4H)

[0215] 1.3 Synthetic Chemical Formula 1-3

[0216] The following reaction formula 3 shows the synthesis process of the thiobenzimidazole derivatives of Chemical Formula 1-3.

[0217] [Reaction formula 3]

[0218]

[0219] 1.3.1. Synthesis of (5-((4-(4-methylpiperazin-1-yl)phenyl)thio)-2-nitrophenyl)carbamate

[0220] To a solution of tert-butyl (5-((4-(4-bromophenyl)thio)-2-nitrophenyl)carbamate (9.0 g, 21.2 mmol, 1.0 eq) in tolane were added 1-methylpiperazine (4.2 g, 42.4 mmol, 2.0 eq) and Pd2(dba)3 (1.9 g, 2.12 mmol, 0.1 eq) in sequence. Xantphos (2.0 g, 4.24 mmol, 0.2 eq) and t-BuONa (4.1 g, 42.4 mmol, 2.0 eq) were added, and the mixture was stirred at 100°C for 16 hours. After completion of the reaction as confirmed by LCMS, the reactant was extracted with DCM, and the organic layer was concentrated and purified by TLC to give tert-butyl (5-((4-(4-methylpiperazin-1-yl)phenyl)thio)-2-nitrophenyl)carbamate (4 g, 43% yield) as a yellow solid.

[0221] 1.3.2. Synthesis of 5-((4-(4-methylpiperazin-1-yl)phenyl)thio)-2-nitroaniline

[0222] A solution of (5-((4-(4-methylpiperazin-1-yl)phenyl)thio)-2-nitrophenyl)carbamate (3 g, 6.75 mmol, 1.0 equiv) in HCl / dioxane (30 mL) was stirred at room temperature for 2 hours, and the reaction was confirmed to be complete by LCMS. NaHCO 3 was added to the mixture, and the mixture was extracted with DCM. After concentration of the organic layer, purification by TLC gave 5-((4-(4-methylpiperazin-1-yl)phenyl)thio)-2-nitroaniline (2.3 g, 95% yield) as a yellow solid.

[0223] 1.3.3. Preparation of 4-((4-(4-methylpiperazin-1-yl)phenyl)thio)benzene-1,2-diamine

[0224] To a solution of 5-((4-(4-methylpiperazin-1-yl)phenyl)thio)-2-nitroaniline (2.3 g, 6.68 mmol, 1.0 eq) in EtOH (50 mL) and H₂O (10 mL) were added Fe (1.8 g, 33.4 mmol, 5.0 eq) and NH₄Cl (3.7 g, 66.8 mmol, 5.0 eq). The mixture was stirred at 80° C. for 2 hours, and the reaction was confirmed to be complete by LCMS. The reactant was then extracted with DCM, and the organic layer was concentrated and purified by TLC to give 4-((4-(4-methylpiperazin-1-yl)phenyl)thio)benzene-1,2-diamine (1.9 g, 90% yield) as a brown solid.

[0225] 1 H NMR (400MHz, DMSO-d6): δ7.03(d,J=8.8Hz,2H),6.84(d,J=8.8Hz,2H),6.56(s,1H) ,6.46(s,2H),4.57-4.66(m,4H),3.06-3.08(m,4H),2.40-2.42(m,4H),2.19(s,1H)

[0226] 1.3.4. Preparation of (5-((4-(4-methylpiperazin-1-yl)phenyl)thio)-1H-benzo[d]imidazol-2-yl)carbamate

[0227] To a solution of 4-((4-(4-methylpiperazin-1-yl)phenyl)thio)benzene-1,2-diamine (800 mg, 2.54 mmol, 1.0 equiv) in CH3COOH (20 mL) was added 1,3-bis(methoxycarbonyl)-S-methylisothiourea and the mixture was stirred at 80 ° C for 2 hours, after which the reaction was confirmed to be complete by LCMS. The resulting mixture was dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated and an aliquot was purified by Prep-TLC to give methyl (5-((4-(4-methylpiperazin-1-yl)phenyl)thio)-1H-benzo[d]imidazol-2-yl)carbamate (632 mg, 63% yield) as a white solid.

[0228] 1H NMR(400MHz,DMSO-d6)δ7.28-7.32(m,2H),7.20(d,J=8.8Hz,2H),6.99-7.01(m,1H), 6.91(d,J=8.8Hz,2H),3.71(s,3H),3.11-3.14(m,4H),2.41-2.43(m,4H),2.20(s,3H)

[0229] 1.4 Synthetic Chemical Formula 1-4

[0230] The following reaction formula 4 shows the synthesis process of the thiobenzimidazole derivatives of Chemical Formula 1-4.

[0231] [Reaction formula 4]

[0232]

[0233] 1.4.1. Synthesis of methyl (5-thiocyanato-1H-benzo[d]imidazol-2-yl) carbamate

[0234] To a solution of 2-nitro-4-thiocyanatoaniline (1.95 g, 10.0 mmol) in AcOH (50 mL) was added zinc powder (5.00 g, 70.0 mmol) and 1,3-bis(methoxycarbonyl)-2-methyl-2-thioisourea (2.70 g, 13.0 mmol). The mixture was stirred at 90 ° C for 20 minutes. After the reaction was complete, the mixture was filtered. The organic layer was then concentrated in vacuo. The residue was purified by reverse phase column chromatography (CH3CN / H2O) to give 300 mg of methyl (5-thiocyanato-1H-benzo [d] imidazole-2-yl) carbamate as a yellow solid.

[0235] 1.4.2. Synthesis of methyl (5-(phenyldisulfanyl)-1H-benzo[d]imidazol-2-yl) carbamate

[0236] PhSNa (319 mg, 2.40 mmol) and KOH (101 mg, 1.80 mmol) were added to a solution of methyl (5-thiocyanato-1H-benzo [d] imidazole-2-yl) carbamate (300 mg, 1.20 mmol) in EtOH (120 mL). The mixture was stirred at room temperature for 16 hours. After the reaction was completed, EtOH was removed. The residue was purified by prep-HPLC to obtain 90 mg of the compound of chemical formula 1-4 as an off-white solid (TFA salt, yield 22%).

[0237] 1 H-NMR (400MHz, DMSO-d6): δ7.64(s,1H),7.55-7.53(m,2H),7.46-7.44(d,1H),7.42-7.74(m,2H),7.34-7.30(m,2H),3.79(s,3H)

[0238] 1.5 Synthetic Chemical Formula 1-5

[0239] The following reaction formula 5 shows the synthesis process of the thiobenzimidazole derivatives of Chemical Formula 1-5.

[0240] [Reaction formula 5]

[0241]

[0242] 1.5.1. Synthesis of 5-(benzylthio)-2-nitroaniline

[0243] To a solution of phenylmethanethiol (2.40 g, 20.0 mmol) in DMSO (150 mL) were added 5-chloro-2-nitroaniline (3.40 g, 20.0 mmol) and Cs2CO3 (13.0 g, 40.0 mmol), and the mixture was stirred at 80°C for 16 hours. After completion of the reaction, the mixture was poured into water (700 mL) and extracted with EA (400 mL*3). After drying over Na2SO4, the mixture was concentrated and purified by reverse phase column chromatography (ACN / H2O) to give 6.00 g of 5-(benzylthio)-2-nitroaniline as a red oil.

[0244] 1.5.2. Synthesis of 4-(benzylthio)benzene-1,2-diamine

[0245] To a solution of 5-(benzylthio)-2-nitroaniline (2.00 g, 7.70 mmol) in AcOH (50 mL) was added Zn (4.00 g, 61.0 mmol). The mixture was stirred at 90° C. for 3 hours. After the reaction was complete, the mixture was filtered, and the filtrate was concentrated to give 2.20 g of 4-(benzylthio)benzene-1,2-diamine as a brown oil.

[0246] 1.5.3. Preparation of methyl (5-(benzylthio)-1H-benzo[d]imidazol-2-yl) carbamate

[0247] To a solution of 4-(benzylthio)benzene-1,2-diamine (2.00 g, 8.70 mmol) in AcOH (40 mL) was added 1,3-bis(methoxycarbonyl)-2-methyl-2-thioisourea (1.50 g, 7.20 mmol). The mixture was stirred at 85 ° C for 16 hours. After the reaction was completed, AcOH was removed and extracted with EA (50 mL * 3). The organic layer was dried over Na2SO4 and concentrated. The crude product was purified by prep-HPLC to obtain 75 mg of (5-(benzylthio)-1H-benzo[d]imidazol-2-yl)carbamate (Chemical Formula 1-5, yield 11%) as a yellow solid.

[0248] 1 H-NMR (400MHz, DMSO-d6): δ11.61(s,1H),7.38(s,1H),7.32-7.30(m,1H),7.2 6-7.25(m,1H),7.23-7.19(m,4H),7.10-7.07(m,1H),4.14(s,2H),3.75(s,3H)

[0249] 1.6 Synthetic Chemical Formula 1-6

[0250] The following reaction formula 6 shows the synthesis process of the thiobenzimidazole derivatives of Chemical Formula 1-6.

[0251] [Reaction formula 6]

[0252]

[0253] 1.6.1. Synthesis of Methyl 3-((4-(piperidin-1-yl)phenyl)thio)propanoate

[0254] To a solution of 1-(4-bromophenyl)piperidine (5.0 g, 20.83 mmol, 1.0 eq) in anhydrous dioxane (50 ml) was added methyl 3-mercaptopropanoate (16.13 mL, 145.81 mmol, 7.0 eq) and Xantphos (2.4 g, 4.17 mmol, 0.2 eq). N,N-diisopropylethylamine (DIEA, 10.9 mL, 62.49 mmol, 3.0 eq) and Pd2(dba)3 (1.9 g, 2.08 mmol, 0.1 eq) were then added to the mixture, and the reaction was stirred at 110°C under nitrogen for 16 hours. After the reaction was complete, the mixture was cooled to room temperature. The reaction was filtered and the filtrate was concentrated. The residue was purified by silica gel column chromatography (HE:EtOAC=40:1) to give methyl 3-((4-(piperidin-1-yl)phenyl)thio)propanoate (5.9 g, yield 100%) as a yellow oil.

[0255] 1.6.2. Synthesis of Sodium 4-(piperidin-1-yl)benzenethiolate

[0256] To a solution of methyl 3-((4-(piperidin-1-yl)phenyl)thio)propanoate (3.6 g, 12.9 mmol, 1.0 equiv) in anhydrous THF (60 mL) at room temperature was added 20% NaOEt (6.59 g, 19.355 mmol, 1.5 equiv). The reaction was heated at room temperature for 1 hour. After completion of the reaction, the mixture was concentrated to afford a brown compound, sodium 4-(piperidin-1-yl)benzenethiolate (3.45 g, crude product).

[0257] 1.6.3. Synthesis of 2-Nitro-5-((4-(piperidin-1-yl)phenyl)thio)aniline

[0258] A solution of sodium 4-(piperidin-1-yl)benzenethiolate (3.45 g, 16.05 mmol, 1.0 equiv), 4-fluoro-2-di(tert-butoxycarbonyl)nitroaniline (5.98 g, 16.05 mmol, 1.0 equiv), and KCO (6.65 g, 48.15 mmol, 3.0 equiv) in anhydrous 1-methyl-2-pyrrolidinone (NMP, 60 mL) was filled with nitrogen and stirred at 130° C. for 16 hours. After completion of the reaction, the residue was purified by silica gel column chromatography (HE:EtOAC = 20:1) to provide 2-nitro-5-((4-(piperidin-1-yl)phenyl)thio)aniline (630 mg) as a yellow solid.

[0259] 1.6.4. Synthesis of 4-((4-(Piperidin-1-yl)phenyl)thio)benzene-1,2-diamine

[0260] To a solution of 2-nitro-5-((4-(piperidin-1-yl)phenyl)thio)aniline (1-4, 450 mg, 1.37 mmol, 1.0 eq) and Pd / C (110 mg) in MeOH (30 mL) was added MeOH / NH3 (10 mL), followed by stirring at room temperature under a stream of hydrogen (H2) at atmospheric pressure for 18 hours. The reaction mixture was then filtered and the filtrate was concentrated to afford 4-((4-(piperidin-1-yl)phenyl)thio)benzene-1,2-diamine (370 mg, 90.5% yield) as a purple solid.

[0261] 1 H NMR (400MHz, CDCl3): δppm 7.21-7.25(m,2H),6.85(d,J=6.8Hz,2H),6.69-6.74(m,2H),6.62(d,J=8.0Hz,1H),3.34-3.43(m,4H),3.14-3.16(m,4H),1.55-1.69(m,6H)

[0262] 1.6.5. Synthesis of (5-((4-(Piperidin-1-yl)phenyl)thio)-1H-benzo[d]imidazol-2-yl)carbamate

[0263] To a solution of compound 4-((4-(piperidin-1-yl)phenyl)thio)benzene-1,2-diamine (100 mg, 0.334 mmol, 1.0 equiv) in acetic acid (5 mL) was added 1,3-bis(methoxycarbonyl)-S-methylisothiourea (76 mg, 0.367 mmol, 1.1 equiv), and the mixture was stirred at 80 ° C for 2 hours. The reactant was extracted with DCM and washed with saturated NaHCO 3. The organic layer was dried over anhydrous Na 2 SO 4 and filtered. The filtrate was concentrated and the resulting mixture was purified by Prep-TLC to give methyl (5-((4-(piperidin-1-yl)phenyl)thio)-1H-benzo[d]imidazol-2-yl)carbamate (Chemical Formula 1-6, 20 mg, yield 15.7%) as a white solid.

[0264] 1 H NMR (400MHz, DMSO-d6): δ7.33(d,J=8.4Hz,1H),7.28(d,J=1.2Hz,1H),7.20(d,J=8.8Hz,2H),7.01-7 .03(m,1H),6.91(d,J=8.8Hz,2H),6.08(brs,2H),3.74(s,3H),3.11-3.19(m,4H),1.53-1.59(m,6H)

[0265] 1.7 Synthetic Chemical Formula 1-7

[0266] The following reaction formula 7 shows the synthesis process of the thiobenzimidazole derivatives of Chemical Formula 1-7.

[0267] [Reaction formula 7]

[0268]

[0269] 1.7.1. Synthesis of methyl 3-((4-morpholinophenyl)thio)propanoate

[0270] To a solution of 4-(4-bromophenyl)morpholine (7.0 g, 30.9 mmol, 1.0 eq) in anhydrous dioxane (50 ml) were added methyl 3-mercaptopropionate (18.6 g, 154.9 mmol, 5.0 eq) and Xantphos (3.6 g, 6.2 mmol, 0.2 eq). DIEA (12.2 g, 92.9 mmol, 3.0 eq) and Pd2(dba)3 (1.4 g, 1.6 mmol, 0.05 eq) were added sequentially, and the reaction was stirred at 110°C for 16 hours. After completion of the reaction, the mixture was cooled to room temperature. The reaction mixture was filtered and the filtrate was concentrated. The residue was isolated and purified by silica gel column chromatography (HE:EtOAC = 8:1) to afford methyl 3-((4-morpholinophenyl)thio)propionate (7.5 g, 91.2% yield) as a yellow oil.

[0271] 1.7.2. Synthesis of sodium 4-morpholinobenzenethiolate

[0272] To a solution of methyl 3-((4-morpholinylphenyl)thio)propanoate (7.4 g, 26.3 mmol, 1.0 eq) in anhydrous THF (100 mL) was added 20% NaOEt (10.7 g, 31.6 mmol, 1.5 eq) at room temperature. The reaction was allowed to react at room temperature for 1 hour. After completion of the reaction, the mixture was concentrated to afford sodium 4-morpholinylthiophenolate (5.2 g, 88.9% yield) as a brown oil.

[0273] 1.7.3. Synthesis of 4-morpholino-2-nitroaniline

[0274] To a solution of sodium 4-morpholinothiophenolate (5.2 g, 23.9 mmol, 1.0 equiv) in anhydrous 1-methyl-2-pyrrolidinone (60 mL) under nitrogen atmosphere were added 5-fluoro-2-nitroaniline (4.5 g, 28.7 mmol, 1.2 equiv) and KCO (9.9 g, 71.8 mmol, 3.0 equiv), followed by stirring at 130° C. for 16 hours. After completion of the reaction, the residue was purified by silica gel column chromatography (HE:EtOAC = 20:1) to afford 4-morpholino-2-nitroaniline (1.2 g, 15.2% yield) as a yellow solid.

[0275] 1.7.4. Synthesis of 4-morpholinobenzene-1,2-diamine

[0276] To a solution of 4-morpholino-2-nitroaniline (1.2 g, 3.6 mmol, 1.0 equiv) in MeOH (20 mL) was added Pd / C (300 mg), and the mixture was stirred at room temperature under hydrogen (H2) for 18 hours. The reaction was filtered to afford 4-morpholinobenzene-1,2-diamine (640 mg, 60.1% yield) as a purple solid.

[0277] 1 H NMR (400MHz, DMSO-d6): δ7.85(d,J=9.2Hz,1H),7.39-7.44(m,3H),7.06(d,J=8.8Hz ,2H),6.52(d,J=1.6Hz,1H),6.24-6.27(m,1H),3.69-3.76(m,4H),3.19-3.23(m,4H)

[0278] 1.7.5. Synthesis of methyl (5-((4-morpholinophenyl)thio)-1H-benzo[d]imidazol-2-yl) carbamate

[0279] To a solution of 4-morpholinylbenzene-1,2-diamine (100 mg, 0.332 mmol, 1.0 eq) in acetic acid (5 mL) was added 1,3-bis(methoxycarbonyl)-S-methylisothiourea (75 mg, 0.365 mmol, 1.1 eq), and the mixture was stirred at 80 ° C for 2 hours. The reaction was confirmed to be complete by LCMS. The reactant was extracted with DCM and washed with saturated bicarbonate water. The organic layer was dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated and the substance was purified by Prep-TLC to obtain the product (chemical formula 1-7, 30 mg, yield 23.6%) as a white solid.

[0280] 1 H NMR (400MHz, DMSO-d6): δppm 7.34(d,J=8.4Hz,1H),7.30(s,1H),7.23(d,J=8.8Hz,2H),7.02-7.05(m,1H ),6.93(d,J=8.8Hz,2H),3.74(s,3H),3.71-3.73(m,4H),3.10-3.12(m,4H)

[0281] 1.8 Synthetic Chemical Formula 1-8

[0282] The following reaction formula 8 shows the synthesis process of the thiobenzimidazole derivatives of Chemical Formula 1-8.

[0283] [Reaction formula 8]

[0284]

[0285] 1.8.1. Synthesis of 1-(4-bromophenyl)-4-ethylpiperazine

[0286] To a solution of 1-(4-bromophenyl)-4-ethylpiperazine (10.0 g, 41.83 mmol, 1.0 eq) in anhydrous acetonitrile (50 mL) was added ethyl iodide (4.0 mL, 49.78 mmol, 1.2 eq) and KCO (11.47 g, 82.96 mmol, 2.0 eq). After the addition was complete, the reaction was stirred at 70°C under N for 2 hours. The reaction mixture was filtered and the filtrate was concentrated. The residue was purified by silica gel column chromatography (HE:EtOAC = 5:1) to afford 1-(4-bromophenyl)-4-ethylpiperazine (9.84 g, 84.2% yield) as a white solid.

[0287] 1.8.2. Synthesis of methyl 3-((4-(4-ethylpiperazin-1-yl)phenyl)thio)propanoate

[0288] To a solution of 1-(4-bromophenyl)-4-ethylpiperazine (9.4 g, 34.9 mmol, 1.0 eq) in anhydrous dioxane (50 mL) were added methyl 3-mercaptopropionate (27.1 mL, 244.4 mmol, 7.0 eq) and Xantphos (4.04 g, 6.98 mmol, 0.2 eq). Following the addition of DIEA (18.3 mL, 104.7 mmol, 3.0 eq) and Pd(dba) (3.2 g, 3.5 mmol, 0.1 eq), the reaction was stirred at 110°C under a stream of N for 16 hours. The reaction mixture was filtered and the concentrate was concentrated. The residue was purified by silica gel column chromatography (Hexane:EtOAC = 2:1) to afford methyl 3-((4-(4-ethylpiperazin-1-yl)phenyl)thio)propanoate (9.5 g, 88.29% yield) as a yellow oil.

[0289] 1.8.3. Synthesis of sodium 4-(4-ethylpiperazin-1-yl)benzenethiolate

[0290] To a solution of 3-((4-(4-ethylpiperazin-1-yl)phenyl)thio)propanoate (2.81 g, 9.1 mmol, 1.0 eq) in anhydrous THF (60 mL) was added 20% sodium ethoxide (4.64 g, 13.65 mmol, 1.5 eq), and the reaction was then heated to room temperature over 20 minutes. After completion of the reaction, the mixture was concentrated to afford sodium 4-(4-ethylpiperazin-1-yl)benzenethiolate (2.8 g) as a brown solid.

[0291] 1.8.4. Synthesis of 5-((4-(4-ethylpiperazin-1-yl)phenyl)thio)-2-nitroaniline

[0292] Sodium 4-(4-ethylpiperazin-1-yl)benzenethiolate (2.8 g, 11.5 mmol, 1.0 equivalent) was dissolved in anhydrous N-methylpyrrolidine dimethyl ether (60 mL), followed by the addition of tert-butyl(tert-butyloxycarbonyl)(5-chloro-2-nitrophenyl)carbamate (4.29 g, 11.5 mmol, 1.0 equivalent) and K2CO3 (4.77 g, 34.5 mmol, 3.0 equivalent). The solution was stirred at 130°C under a stream of N2 for 16 hours. After completion of the reaction, the residue was purified by silica gel column chromatography (DCM:MeOH = 20:1) to give 5-((4-(4-ethylpiperazin-1-yl)phenyl)thio)-2-nitroaniline (520 mg) as an orange solid.

[0293] 1.8.5. Synthesis of 4-((4-(4-ethylpiperazin-1-yl)phenyl)thio)benzene-1,2-diamine

[0294] To a solution of 5-((4-(4-ethylpiperazin-1-yl)phenyl)thio)-2-nitroaniline (520 mg, 1.45 mmol, 1.0 equiv) in MeOH (15 mL) and MeOH / NH₃ (5 mL) was added Pd / C (150 mg), and the solution was stirred at room temperature under hydrogen (H₂) for 18 hours. The reaction mixture was filtered, and the filtrate was concentrated to afford 4-((4-(4-ethylpiperazin-1-yl)phenyl)thio)benzene-1,2-diamine (470 mg, 99.5% yield) as a purple solid.

[0295] 1 H NMR (400MHz, CDCl3): δ7.22(d,J=8.4Hz,2H),6.82(d,J=8.8Hz,2H),6.73-6.78(m,2H),6.64 (d,J=7.6Hz,1H),3.26-3.46(m,8H),2.78-2.92(m,4H),1.51-1.64(m,2H),1.29-1.30(m,3H)

[0296] 1.8.6. Synthesis of Methyl(5-((4-(4-ethylpiperazin-1-yl)phenyl)thio)-1H-benzo[d]imidazol-2-yl)carbamate

[0297] To a solution of 4-((4-(4-ethylpiperazine-1-yl)phenyl)thio)benzene-1,2-diamine (100 mg, 0.305 mmol, 1.0 equiv) in acetic acid (5 mL) was added 1,3-bis(methoxycarbonyl)-S-methylisothiourea (69 mg, 0.335 mmol, 1.1 equiv), the mixture was stirred at 80 ° C for 2 hours, and the reaction was confirmed to be complete by LCMS. The reactant was extracted with DCM and washed with saturated NaHCO3. The filtrate was concentrated and the product was purified by Prep-TLC to give methyl 5-((4-(4-ethylpiperazine-1-yl)phenyl)thio))-1H-benzo[d]imidazol-2-yl)carbamate (Chemical Formula 1-8, 44 mg, 32% yield) as a white solid.

[0298] 1H NMR (400MHz, DMSO-d6) δ7.33(d,J=8.0Hz,1H),7.29(s,1H),7.21(d,J=8.8Hz,2H),7.00-7.03(m,1H),6.92 (d,J=8.8Hz,2H),3.74(s,3H),3.13-3.15(m,4H),2.46-2.48(m,4H),2.32-2.37(m,2H),1.01-1.04(m,3H)

[0299] 1.9 Synthetic Chemical Formula 1-9

[0300] The following reaction formula 9 shows the synthesis process of the thiobenzimidazole derivatives of Chemical Formula 1-9.

[0301] [Reaction formula 9]

[0302]

[0303] 1.9.1. Synthesis of 1-(3-bromophenyl)-4-methylpiperazine

[0304] To a solution of 1,3-dibromobenzene (9.0 g, 38.5 mmol, 1.0 eq) in toluene (100 mL) were added 1-methylpiperazine (19.2 g, 192.4 mmol, 5.0 eq), DBU (17.5 g, 115.5 mmol, 3.0 eq), and Pd2(dba)3 (2.2 g, 1.9 mmol, 0.05 eq). The reaction mixture was heated at 60°C under N2 with stirring for 16 hours, and then concentrated. The residue was purified by silica gel column chromatography (HE:EtOAC = 60:1) to afford 1-(3-bromophenyl)-4-methylpiperazine (5.5 g, 56.3% yield) as a yellow oil.

[0305] 1.9.2. Synthesis of methyl 3-((3-(4-methylpiperazin-1-yl)phenyl)thio)propanoate

[0306] To a solution of 1-(3-bromophenyl)-4-methylpiperazine (5.5 g, 21.65 mmol, 1.0 equiv) in anhydrous dioxane (50 ml) was added methyl 3-mercaptopropionate (18.2 mg, 155.55 mmol, 7.0 equiv). Xantphos (2.5 g, 4.33 mmol, 0.2 equiv), DIEA (8.4 g, 64.95 mmol, 3.0 equiv), and Pd2(dba)3 (2.0 g, 2.165 mmol, 0.1 equiv) were added sequentially. After the addition was complete, the reaction was stirred at 110°C under N2 for 24 hours. The reaction was filtered and the filtrate was concentrated. The residue was purified by silica gel column chromatography (DCM:MeOH=20:1) to give methyl 3-((3-(4-methylpiperazin-1-yl)phenyl)thio)propanoate (4.5 g, yield 70.7%) as an orange oil.

[0307] 1.9.3. Synthesis of sodium 3-(4-methylpiperazin-1-yl)benzenethiolate

[0308] To a solution of methyl 3-((3-(4-methylpiperazin-1-yl)phenyl)thio)propanoate (2.3 g, 7.82 mmol, 1.0 equiv) in anhydrous THF (60 mL) was added 20% NaOEt (3.8 g, 11.73 mmol, 1.5 equiv). The room temperature reaction was heated for 10 minutes. After completion of the reaction, the mixture was concentrated to afford sodium 3-(4-methylpiperazin-1-yl)benzenethiolphenolate (2.05 g) as a brown solid.

[0309] 1.9.4. Synthesis of 5-((3-(4-methylpiperazin-1-yl)phenyl)thio)-2-nitroaniline

[0310] To a solution of sodium 3-(4-methylpiperazin-1-yl)benzenethiolate (2.05 g, 8.91 mmol, 1.0 equiv) in anhydrous N-methylpyrrolidine dimethyl ether was added tert-butyl(tert-butyloxycarbonyl)(5-chloro-2-nitrophenyl)carbamate (3.3 g, 8.91 mmol, 1.0 equiv) and KCO (3.7 g, 26.73 mmol, 3.0 equiv) in sequence, and the mixture was stirred at 130° C. under N for 16 hours. After completion of the reaction, the residue was purified by silica gel column chromatography (DCM:MeOH=20:1) to afford 5-((3-(4-methylpiperazin-1-yl)phenyl)thio)-2-nitroaniline (3.1 g) as an orange solid.

[0311] 1.9.5. Synthesis of 4-((3-(4-methylpiperazin-1-yl)phenyl)thio)benzene-1,2-diamine

[0312] To a solution of 5-((3-(4-methylpiperazin-1-yl)phenyl)thio)-2-nitroaniline (3.1 g, 9.0 mmol, 1.0 equiv) and Pd / C (1.0 g) in MeOH (60 mL) was added MeOH / NH (20 mL) and stirred under H for 18 hours. The reaction was filtered and the filtrate was concentrated to afford 4-((3-(4-methylpiperazin-1-yl)phenyl)thio)benzene-1,2-diamine (600 mg) as a gray solid.

[0313] 1 H NMR (400MHz, CDCl3): δ7.07-7.12(m,1H),6.84-6.89(m,2H),6.78-6.80(m,1H),6.66-6.70(m,2H),6. 60-6.63(m,1H),3.49(s,2H),3.36(s,2H),3.14-3.17(m,4H),2.52-2.55(m,4H),2.34(d,J=5.6Hz,3H)

[0314] 1.9.6. Synthesis of methyl (5-((3-(4-methylpiperazin-1-yl)phenyl)thio)-1H-benzo[d]imidazol-2-yl)carbamate

[0315] To a solution of 4-((3-(4-methylpiperazin-1-yl)phenyl)thio)benzene-1,2-diamine (100 mg, 0.318 mmol, 1.0 equiv) in CH3COOH (10 mL) was added 1,3-bis(methoxycarbonyl)-S-methylisothiourea (72 mg, 0.35 mmol, 1.1 eq). The reaction mixture was stirred at 80 ° C for 2 hours, and the reaction was confirmed to be complete by LCMS. The reactants were extracted with DCM and dried over anhydrous Na2SO4 and the sat.NaHCO3 organic layer was filtered. The filtrate was concentrated and the mixture was purified by Prep-TLC to obtain methyl (5-((3-(4-methylpiperazin-1-yl)phenyl)thio)-1H-benzo[d]imidazol-2-yl)carbamate (Chemical Formula 1-9, 20 mg, yield 15.87%) as a white solid.

[0316] 1 H NMR (400MHz, DMSO-d6) δ7.49 (s, 1H), 7.43 (d, J = 8.0Hz, 1H), 7.16-7.18 (m, 1H), 7.08-7.12 (m, 1H), 6.7 7(d,J=6Hz,2H),6.47(d,J=7.6Hz,1H),3.76(s,3H),3.04-3.07(m,4H),2.38-2.40(m,4H),2.19(s,3H)

[0317] 1.10 Synthetic Chemical Formula 1-10

[0318] The following reaction formula 10 shows the synthesis process of the thiobenzimidazole derivative of Chemical Formula 1-10.

[0319] [Reaction formula 10]

[0320]

[0321] 1.10.1. Synthesis of tert-butyl(tert-butoxycarbonyl)(5-chloro-2-nitrophenyl)carbamate

[0322] To a solution of 5-chloro-2-nitroaniline (20 g, 116.2 mmol, 1.0 equiv) in THF (300 mL) was added di-tert-butyl dicarbonate (50 g, 232.4 mmol, 2.0 equiv) and DMAP (14 g, 116.2 mmol, 1.0 equiv). The reaction mixture was stirred at 70°C for 1 hour. LCMS confirmed the reaction was complete. The reaction was concentrated and purified by silica gel column chromatography (HE:EA = 10:1) to afford tert-butyl(tert-butoxycarbonyl)(5-chloro-2-nitrophenyl)carbamate (36 g, 85% yield) as a yellow solid.

[0323] 1.10.2. Synthesis of tert-butyl(5-((4-bromophenyl)thio)-2-nitrophenyl)carbamate

[0324] To tert-butyl (tert-butyloxycarbonyl) (5-chloro-2-nitrophenyl) carbamate (28g, 75.2mmol, 1.0 equivalent) DMF (100mL) solution was added 4-bromobenzenethiol (21g, 112.9mmol, 1.5eq) and KCO (20.7g, 150.4mmol, 2eq) in sequence, and the reaction mixture was stirred at 100°C for 16 hours. The reaction was confirmed to be complete by LCMS. The reactant was extracted with DCM and washed with salt water. The organic layer was dried and concentrated to give tert-butyl (5-((4-bromophenyl) sulfenyl)-2-nitrophenyl) carbamate (10.2g, 35% yield) as a yellow solid.

[0325] 1.10.3. Synthesis of tert-butyl(2-nitro-5-((4-(thiophen-3-yl)phenyl)thio)phenyl)carbamate

[0326] To a solution of tert-butyl (5-((4-bromophenyl)thio)-2-nitrophenyl)carbamate (4.3 g, 10.1 mmol, 1.0 equiv) in DMF (20 mL) and H₂O (4 mL) was added thiophen-3-ylboronic acid (2.6 g, 20.2 mmol, 2.0 equiv). Pd(PPh₃)₄ (1.2 g, 1.01 mmol, 0.1 equiv) and K₂CO₃ (4.2 g, 30.3 mmol, 3.0 equiv) were added sequentially, followed by stirring at 100°C for 16 hours. After completion of the reaction as confirmed by LCMS, the reaction was extracted with DCM and washed with brine. The organic layer was dried over anhydrous Na₂SO₄ and filtered. The filtrate was concentrated and the residue was purified by silica gel column chromatography (elution with HE:EA=3:1) to give tert-butyl(2-nitro-5-((4-(thiophen-3-yl)phenyl)thio)phenyl)carbamate (2.7 g, yield 62%) as a yellow solid.

[0327] 1.10.4. Synthesis of 2-nitro-5-((4-(thiophen-3-yl)phenyl)thio)aniline

[0328] A solution of tert-butyl (2-nitro -5- ((4- (thiophene-3-yl) phenyl) thio) phenyl) carbamate (2.6 g, 6.07 mmol, 1.0 equivalent) in HCl / dioxane (20 mL) was stirred at room temperature for 2 hours. After completion of the reaction, the reaction mixture was poured into 100 mL of NaHCO 3 ice solution. The mixture was extracted with DCM and washed with brine. The organic layer was dried over anhydrous Na 2 SO 4 and filtered. The filtrate was concentrated and purified by silica gel column chromatography eluting with DCM: MeOH = 10: 1 to give 2-nitro -5- ((4- (thiophene-3-yl) phenyl) thio) aniline (1.5 g, 78% yield) as a yellow solid.

[0329] 1.10.5. Synthesis of 4-((4-(thiophen-3-yl)phenyl)thio)benzene-1,2-diamine

[0330] To a solution of 2-nitro-5-((4-(thiophen-3-yl)phenyl)thio)aniline (1.3 g, 3.96 mmol, 1.0 equiv) in EtOH (20 mL) and H₂O (5 mL) were added Fe (1.1 g, 19.8 mmol, 5.0 eq) and NHCl (1.1 g, 19.8 mmol, 5.0 eq), and the mixture was stirred at 80°C for 2 hours. After the reaction was complete and monitored by LCMS, the mixture was extracted with DCM and washed with brine. The organic layer was dried over anhydrous Na₂SO₄ and filtered. The filtrate was concentrated and purified by silica gel column chromatography eluting with DCM:MeOH = 10:1 to afford 4-((4-(thiophen-3-yl)phenyl)thio)benzene-1,2-diamine (900 mg, 76% yield) as a brown solid.

[0331] 1 H NMR(400MHz,DMSO-d6)δ7.77-7.78(m,1H),7.57-7.76(m,3H),7.47-7.49(m ,1H),7.03-7.06(m,2H),6.68(s,1H),6.56-6.61(m,2H),4.67-4.84(m,4H)

[0332] 1.10.6. Synthesis of methyl (5-((4-(thiophen-3-yl)phenyl)thio)-1H-benzo[d]imidazol-2-yl) carbamate

[0333] To a solution of 4-((4-(thiophen-3-yl)phenyl)thio)benzene-1,2-diamine (100 mg, 0.335 mmol, 1.0 equiv) in CH3COOH (5 mL) was added 1,3-bis(methoxycarbonyl)-S-methylisothiourea (76 mg, 0.369 mmol, 1.1 equiv). The reaction mixture was stirred at 80 ° C for 2 hours. After the reaction was completed as confirmed by LCMS, the reaction product was extracted with DCM and washed with a saturated solution. The organic layer was washed with saturated NaHCO3, dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated and purified by Prep-TLC to give methyl (5-((4-(thiophen-3-yl)phenyl)thio)-1H-benzo[d]imidazol-2-yl)carbamate (Chemical Formula 1-10, 50 mg, yield 39.1%) as a white solid.

[0334] 1 H NMR (400MHz, DMSO-d6): δppm 7.82-7.84(m,1H),7.61-7.66(m,3H),7.45-7.54(m,3H),7.10-7.23(m,3H),3.76(s,3H)

[0335] 1.11 Synthetic Chemical Formula 1-11

[0336] The following reaction formula 11 shows the synthesis process of the thiobenzimidazole derivative of Chemical Formula 1-11.

[0337] [Reaction formula 11]

[0338]

[0339] 1.11.1. Synthesis of Methyl 3-(furan-2-ylthio)propanoate

[0340] To a solution of 2-bromofuran (6.0 g, 40.8 mmol, 1.0 eq) in anhydrous dioxane (50 mL) were added methyl 3-mercaptopropanoate (31.6 mL, 285.6 mmol, 7.0 eq), Xantphos (4.787 g, 8.16 mmol, 0.2 eq), DIEA (21.4 mL, 122.4 mmol, 3.0 eq), and Pd2(dba)3 (3.623 g, 4.08 mmol, 0.1 eq). After the addition was complete, the reaction was stirred at 110°C under N2 for 16 hours. The reaction was filtered and the filtrate was concentrated. The obtained product was purified by silica gel column chromatography (HE:EtOAC=40:1) to obtain methyl 3-(furan-2-ylthio)propanoate (6.34 g, yield 83.4%) as a yellow oily liquid.

[0341] 1.11.2. Synthesis of sodium furan-2-thiolate

[0342] To a solution of methyl 3-(furan-2-ylthio)propanoate (6.34 g, 34.08 mmol, 1.0 eq) in anhydrous THF (60 mL) at room temperature was added 20% NaOEt (14.06 g, 41.32 mmol, 1.2 eq). The reaction mixture was heated for 1 hour to complete the reaction, and then the mixture was concentrated to afford sodium furan-2-thiolate (4.61 g) as a brown solid.

[0343] 1.11.3. Synthesis of 5-(furan-2-ylthio)-2-nitroaniline

[0344] A solution of sodium furan-2-thiolate (4.61 g, 37.86 mmol, 1.1 equivalent), 5-fluoro-2-nitroaniline (5.39 g, 34.4 mmol, 1.0 equivalent), and KCO (14.26 g, 103.2 mmol, 3.0 equivalent) in anhydrous N-methylpyrrolidinedimethyl (60 mL) was stirred at 130° C. under N for 16 hours. After completion of the reaction, the residue was purified by silica gel column chromatography (HE:EtOAC = 20:1) to afford 5-(furan-2-ylthio)-2-nitroaniline (860 mg) as a yellow solid.

[0345] 1.11.4. Synthesis of 4-(furan-2-ylthio)benzene-1,2-diamine

[0346] 5-(Furan-2-ylthio)-2-nitroaniline (860 mg, 3.624 mmol, 1.0 equivalent) and Pd / C (360 mg) were added to a mixed solvent of MeOH (36 mL) and MeOH / NH (12 mL) and stirred at room temperature under H for 18 hours. The reaction mixture was filtered and the filtrate was concentrated to provide 4-(furan-2-ylthio)benzene-1,2-diamine (750 mg, 100% yield) as a purple solid.

[0347] 1 H NMR (400MHz, CDCl3): δppm 7.49-7.50(m,1H),6.69-6.74(m,2H),6.59-6.62(m,2H),6.39-6.41(m,1H),3.37(brs,4H)

[0348] 1.11.5. Synthesis of methyl (5-(furan-2-ylthio)-1H-benzo[d]imidazol-2-yl) carbamate

[0349] To a solution of 4-(furan-2-ylthio)benzene-1,2-diamine (100 mg, 0.485 mmol, 1.0 equivalent) in CH3COOH (5 mL) was added 1,3-bis(methoxycarbonyl)-S-methylisothiourea (100 mg, 0.485 mmol, 1.1 equivalents), and the mixture was stirred at 80 ° C for 2 hours. The reactants were monitored by LCMS to confirm the completion of the reaction, and then extracted with DCM and washed with saturated NaHCO3 solution. The organic layer was dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated and the reaction residue was purified by Prep-TLC to give a white solid compound methyl (5-(furan-2-ylthio)-1H-benzo [d] imidazole-2-yl) carbamate (chemical formula 1-11, 20 mg, yield 14.2%).

[0350] 1 H NMR (400MHz, DMSO-d6): δppm 7.88-7.89(m,1H),7.40(d,J=8.4Hz,1H),7.30(d,J=8.4Hz,1H),7.07-7.10(m,1H),6.92-6.93(m,1H),6.61-6.63(m,1H),3.78(s,3H)

[0351] 1.12 Synthetic Chemical Formula 1-12

[0352] Using 3,4-difluorobenzenethiol as the starting material, methyl (5-((3,4-difluorophenyl)thio)-1H-benzo[d]imidazol-2-yl)carbamate (Chemical Formula 1-12, yield 69.2%) was synthesized according to the same method as Chemical Formula 1-1 in Example 1.1.

[0353] 1 H NMR (CDCl3, 400MHz) 3.76 (s, 3H), 6.93 (m, 1H), 7.17 (m, 2H), 7.19 (d, J = 2.32Hz, 1H),7.23(q,1H),7.38(d,J=10.64Hz),7.55(s,1H),11.53(m,1H),12.02(m,1H)

[0354] 1.13 Synthetic chemical formula 1-13

[0355] Using 2,4-difluorobenzenethiol as the starting material, methyl(5-((2,4-difluorophenyl)thio)-1H-benzo[d]imidazol-2-yl)carbamate (Chemical Formula 1-13, yield 75.4%) was synthesized according to the same method as Chemical Formula 1-1 in Example 1.1.

[0356] 1 H NMR(DMSO-d6,400MHz)3.75(s,3H),7.12(m,3H),7.33(m,1H),7.38(m,1H),7.47(s,1H),11.73(m,2H)

[0357] 1.14 Synthetic chemical formula 1-14

[0358] 5-chloropyridine-2-thiol was used as the starting material and (methyl(5-((5-chloropyridin-2-yl)thio)-1H-benzo[d]imidazol-2-yl)carbamate (Chemical Formula 1-14, yield 75.2%) was synthesized according to the same method as Chemical Formula 1-1 in Example 1.1.

[0359] 1H NMR(DMSO-d6,400MHz)3.77(s,3H),6.75(d,J=8.68Hz,1H),7.31(d,J=1.68Hz), 7.53(d,J=8.2Hz,1H),7.64(s,1H),7.69(d,J=2.6Hz),8.44(s,1H),11.96(m,2H)

[0360] 1.15 Synthetic chemical formula 1-15

[0361] The following reaction formula 15 shows the synthesis process of the thiobenzimidazole derivative of Chemical Formula 1-15.

[0362] [Reaction formula 15]

[0363]

[0364] 1.15.1. Synthesis of 5-((4-Bromophenyl)thio)-2-nitroaniline

[0365] Potassium carbonate (1.24 g) was dissolved in a solution of 4-bromobenzenethiol (1.4 g, 8.7 mmol) in DMF (10 mL). A previously prepared solution of 5-chloro-2-nitroaniline (1.49 g, 8.7 mmol) in DMF (3 mL) was added at room temperature. The reaction mixture was stirred at 90°C for 3 hours, then cooled to room temperature and poured into cold water (500 mL). The precipitate was filtered and dried to obtain 5-((4-bromophenyl)thio)-2-nitroaniline (2.8 g, 99% yield) as a yellow solid.

[0366] 1.15.2. Synthesis of tert-Butyl(5-((4-bromophenyl)thio)-2-nitrophenyl)carbamate

[0367] To a solution of 5-((4-bromophenyl)thio)-2-nitroaniline (2.8 g, 8.7 mmol) in THF (30 mL) was added di-tert-butyl dicarbonate (5.14 g, 23.5 mmol) and DMAP (0.146 g), followed by heating under reflux for 1 hour. After TLC confirmed the completion of the reaction, the reactants were cooled to room temperature and evaporated under reduced pressure to remove the solvent. The product was dissolved in methanol (30 mL) and potassium carbonate (5.2 g) was added, followed by stirring at room temperature for 6 hours to give tert-butyl (5-((4-bromophenyl)thio)-2-nitrophenyl)carbamate (1.48 g, 40% yield).

[0368] 1.15.3. Synthesis of tert-butyl(2-nitro-5-((4-(thiophen-2-yl)phenyl)thio)phenyl)carbamate

[0369] To a solution of tert-butyl (5-((4-bromophenyl)thio)-2-nitrophenyl)carbamate (0.7 g, 1.6 mmol) in DMF (5 mL) were added thiophen-2-ylboronic acid (0.42 g, 3.28 mmol, 2.0 eq), tetrakis(triphenylphosphine)palladium (0.18 g, 0.16 mmol, 0.1 eq) and potassium carbonate (0.66 g, 4.80 mmol, 3 eq), followed by stirring at 100° C. for 15 hours. After cooling to room temperature, the mixture was extracted with methylene chloride solution and washed once with Brine solution, dried over anhydrous magnesium sulfate, evaporated under reduced pressure, and then eluted with ethyl acetate and hexane (7:3, v / v) by silica gel column chromatography. The tert-butyl (2-nitro-5-((4-(thiophene-2-

[0370] 4-[4-amino]-phenyl]-thio]-phenyl]-carbamate (0.48 g, yield 70%).

[0371] 1.15.4. Synthesis of 2-nitro-5-((4-(thiophen-2-yl)phenyl)thio)aniline

[0372] A solution of tert-butyl (2-nitro-5-((4-(thiophen-2-yl)phenyl)thio)phenyl)carbamate (0.48 g, 1.1 mmol) in HCl / dioxane (6 mL) was reacted at room temperature for 2 hours, and then poured into a cold saturated aqueous sodium bicarbonate solution (30 mL). The mixture was extracted with an excess of dichloromethane solution and dried over anhydrous sodium sulfate. The solvent was then removed by filtration and evaporated under reduced pressure. The residue was then purified by silica gel column chromatography to give 2-nitro-5-((4-(thiophen-2-yl)phenyl)thio)aniline (413 mg, yield 78%).

[0373] 1.15.5. Synthesis of 4-((4-(thiophen-2-yl)phenyl)thio)benzene-1,2-diamine

[0374] To a solution of 2-nitro-5-((4-(thiophen-2-yl)phenyl)thio)aniline (413 mg, 1.2 mmol) in ethanol (15 mL) was added tin(II) chloride dihydrate (1.74 g, 7.7 mmol, 6.4 eq), heated under reflux for 3 hours and then cooled to room temperature, followed by addition of water (50 mL). After extraction with an excess of ethyl acetate, the solvent was removed by evaporation under reduced pressure to obtain a solid reaction mixture. It was separated and purified by silica gel column chromatography using ethyl acetate and hexane (1:3, v / v) solvent to obtain 4-((4-(thiophen-2-yl)phenyl)thio)benzene-1,2-diamine (0.20 g, 57% yield).

[0375] 1.15.6. Synthesis of Methyl(5-((4-(thiophen-2-yl)phenyl)thio)-1H-benzo[d]imidazol-2-yl)carbamate

[0376] 4-((4-(thiophen-2-yl)phenyl)thio)benzene-1,2-diamine (0.1 g, 0.33 mmol) and 1,3-bis(methoxycarbonyl)-S-methylisothiourea (0.55 g, 2.6 mmol, 8 eq) were dissolved in 3 mL of 5% AcOH in ethanol and reacted at 80°C for 2 hours. After TLC confirmed the completion of the reaction, extraction was performed with excess dichloromethane, followed by neutralization with saturated sodium bicarbonate and drying over anhydrous sodium sulfate. Methyl (5-((4-(thiophen-2-yl)phenyl)thio)-1H-benzo[d]imidazol-2-yl)carbamate (Chemical Formula 1-15, 90 mg, 72% yield) was obtained as a light brown solid by silica gel column chromatography.

[0377] 1 H NMR(DMSO-d6,400MHz)3.76(s,3H),7.12(m,3H),7.23(d,J=8.16Hz,1H),7.47(m,2H),7.51-7.58(m,4H),11.77(m,2H)

[0378] 1.16 Synthetic chemical formula 1-16

[0379] The following reaction formula 16 shows the synthesis process of the thiobenzimidazole derivatives of Chemical Formula 1-16.

[0380] [Reaction formula 16]

[0381]

[0382] 1.16.1. Preparation of 5-((4-(1H-imidazole-1-yl)phenylthio)-2-nitroaniline

[0383] To a solution of (5-((4-fluorophenyl)thio)-2-nitroaniline (5-((4-fluorophenyl)thio)-2-nitroaniline, 0.85 g, 3.2 mmol) in DMSO (5 mL) was added 1H-imidazole (0.23 g, 3.3 mmol) and stirred for 5 minutes, and then potassium t-butoxide (0.36 g, 3.2 mmol) was slowly added and reacted at 90° C. for 2 hours. The reaction mixture was extracted with ethyl acetate and washed with water, and then evaporated under reduced pressure to remove the solvent to obtain a mixed product. It was separated and purified by silica gel column chromatography using ethyl acetate and hexane (3:1, v / v) solvent to give 5-((4-(1H-imidazol-1-yl)phenylthio)-2-nitroaniline (0.32 g, yield 32%).

[0384] 1.16.2. Preparation of 5-((4-(1H-imidazole-1-yl)phenylthio)-1,2-diamine

[0385] To a solution of 5-((4-(1H-imidazol-1-yl)phenylthio)-2-nitroaniline (0.309 g, 1.0 mmol) in ethanol (10 mL) was added tin chloride dihydrate (6 equivalents) and the mixture was heated under reflux for 4 hours, then cooled to room temperature, and then water (50 mL) was added. After extraction with excess ethyl acetate, the solvent was removed by evaporation under reduced pressure to obtain a solid reaction mixture. This was separated and purified by silica gel column chromatography using ethyl acetate and hexane (3:1, v / v) to obtain 5-((4-(1H-imidazol-1-yl)phenylthio)-1,2-diamine (0.30 g, yield 98%).

[0386] 1.16.3. Synthesis of methyl (5-((4-(1H-imidazol-1-yl)phenyl)thio)-1H-benzo[d]imidazol-2-yl)carbamate

[0387] To a solution of 5-((4-(1H-imidazol-1-yl)phenylthio)-1,2-diamine (0.28 g, 1.0 mmol) in 5% acetic acid in ethanol (3.0 mL) was added 1,3-bis(methoxycarbonyl)-2-methyl-2-thiopseudourea (0.52 g, 2.5 mmol) and heated under reflux for 5 hours. The reaction mixture was filtered and washed thoroughly with methanol to obtain methyl (5-((4-(1H-imidazol-1-yl)phenyl)thio)-1H-benzo[d]imidazol-2-yl)carbamate (Chemical Formula 1-16, 0.26 g, 70% yield).

[0388] 1 H NMR(DMSO-d6,400MHz)3.76(s,3H),7.18(s,1H),7.23(m,3H),7.48(m,1H),7.58(m,3H),7.68(s,1H),8.20(s,1H),11.46(m,1H),11.94(m,1H)

[0389] 1.17 Synthetic chemical formula 1-17

[0390] Methyl(5-((4,6-dimethylpyrimidin-2-yl)thio)-1H-benzo[d]imidazol-2-yl)carbamate (Formula 1-17, yield 75.7%) was synthesized using 4,6-dimethylpyrimidine-2-thiol as the starting material according to the same method as Formula 1-1 in Example 1.1.

[0391] 1 H NMR(DMSO-d6,400MHz)2.25(s,6H),3.77(s,3H),6.93(s,1H),7.26(d,J=8.24Hz,1H),7.45(d,J=8.20Hz,1H),7.61(s,1H),11.74(m,2H)

[0392] 1.18 Synthetic chemical formula 1-18

[0393] Pyridine-2-thiol was used as the starting material, and methyl(5-(pyridin-2-ylthio)-1H-benzo[d]imidazol-2-yl)carbamate (Chemical Formula 1-18, yield 72.5%) was synthesized according to the same method as Chemical Formula 1-1 in Example 1.1.

[0394] 1 H NMR(DMSO-d6,400MHz)3.77(s,3H),6.73(d,J=8.08,1H),7.08(m,1H),7.28(d, J=6.72Hz,1H),7.55(m,2H),7.64(s,1H),8.37(d,J=3.36Hz,1H),11.89(m,2H).

[0395] 1.19 Synthetic chemical formula 1-19

[0396] The following reaction formula 19 shows the synthesis process of the thiobenzimidazole derivative of Chemical Formula 1-19.

[0397] [Reaction formula 19]

[0398]

[0399] 1.19.1. Synthesis of 5-((4-(1H-1,2,4-triazol-1-yl)phenyl)thio)-2-nitroaniline

[0400] 1H-1,2,4-triazole (1H-1,2,4-triazole, 2.2 g, 32 mmol) was added to a DMSO (30 mL) solution of (5-((4-fluorophenyl)thio)-2-nitroaniline (2.0 g, 29 mmol) and stirred for 5 minutes, and then potassium tert-butoxide (3.6 g, 33 mmol) was slowly added, followed by reaction at 90°C for 4 hours. The reaction mixture was extracted with ethyl acetate and washed with water, and then evaporated under reduced pressure to remove the solvent to obtain a mixture. It was separated and purified by silica gel column chromatography using ethyl acetate and hexane (3:1, v / v) solvent to obtain 5-((4-(1H-1,2,4-triazol-1-yl)phenyl)thio)-2-nitroaniline (0.94 g, yield 47%).

[0401] 1.19.2. Synthesis of 4-((4-(1H-1,2,4-triazol-1-yl)phenyl)thio)benzene-1,2-diamine

[0402] To a solution of 5-((4-(1H-1,2,4-triazol-1-yl)phenyl)thio)-2-nitroaniline (0.94 g, 3.0 mmol) in ethanol (100 mL) was added SnCl2·2H2O (6 eq) and heated under reflux for 5 hours, then cooled to room temperature and added with water (300 mL). After extraction with an excess of ethyl acetate, the solvent was removed by evaporation under reduced pressure to obtain a solid reaction mixture. It was separated and purified by silica gel column chromatography using ethyl acetate and hexane (3:1, v / v) solvent to obtain 4-((4-(1H-1,2,4-triazol-1-yl)phenyl)thio)benzene-1,2-diamine (0.85 g, yield 99%).

[0403] 1.19.3. Synthesis of methyl (5-((4-(1H-1,2,4-triazol-1-yl)phenyl)thio)-1H-benzo[d]imidazol-2-yl)carbamate

[0404] To a solution of 4-((4-(1H-1,2,4-triazol-1-yl)phenyl)thio)benzene-1,2-diamine (0.85 g, 3.0 mmol) in 5% acetic acid in ethanol (10 mL) was added 1,3-bis(methoxycarbonyl)-2-methyl-2-thioisourea (1.5 g, 7.7 mmol) and heated under reflux for 5 hours. The reaction mixture was cooled to room temperature and filtered. At this point, the mixture was washed thoroughly with methanol to obtain methyl (5-((4-(1H-1,2,4-triazol-1-yl)phenyl)thio)-1H-benzo[d]imidazol-2-yl)carbamate (Chemical Formula 1-19, 0.81 g, 74% yield).

[0405] 1 H NMR(DMSO-d6,400MHz)3.76(s,3H),7.25(m,1H),7.28(d,J=8.6Hz,2H),7.47(m, 1H),7.57(s,1H),7.78(d,J=8.6Hz,2H),8.21(s,1H),9.23(s,1H),11.80(m,2H)

[0406] 1.20 Synthetic chemical formula 1-20

[0407] Using 4-methyl-1H-imidazole as the starting material, methyl (5-((4-(4-methyl-1H-imidazol-1-yl)phenyl)thio)-1H-benzo[d]imidazol-2-yl)carbamate (Chemical Formula 1-20, yield 78.1%) was synthesized according to the same method as Chemical Formula 1-19 in Example 1.19.

[0408] 1 H NMR(DMSO-d6,400MHz)2.14(s,3H),3.76(s,3H),7.23(d,J=8.4Hz,3H),7.37(s, 1H),7.47(m,1H),7.53(d,J=8.8Hz,3H),8.07(s,1H),11.47(m,1H),12.01(M,1H)

[0409] 1.21 Synthetic chemical formula 1-21

[0410] 1.21 Synthesis of Chemical Formula 1-21

[0411] Using 3,5-dimethyl-1H-1,2,4-triazole (3,5-dimethyl-1H-1,2,4-triazole) as the starting material, methyl (5-((4-(3,5-dimethyl-1H-1,2,4-triazol-1-yl)phenyl)thio)-1H-benzo[d]imidazol-2-yl)carbamate (Chemical Formula 1-21, yield 71.9%) was synthesized according to the same method as Chemical Formula 1-19 in Example 1.19.

[0412] 1 H NMR(DMSO-d6,400MHz)2.24(s,3H),2.37(s,3H),3.76(s,3H),7.20(d,J=8.8Hz,2H),7. 28(m,1H),7.43(d,J=8.8Hz,2H),7.15(m,1H),7.60(s,1H),11.41(m,1H),12.37(m,1H)

[0413] 1.22 Synthetic chemical formula 1-22

[0414] 3-methyl-1H-1,2,4-triazole was used as the starting material and methyl(5-((4-(3-methyl-1H-1,2,4-triazol-1-yl)phenyl)thio)-1H-benzo[d]imidazol-2-yl)carbamate (Chemical Formula 1-22, yield 70.4%) was synthesized according to the same method as Chemical Formula 1-19 in Example 1.19.

[0415] 1 H NMR(DMSO-d6,400MHz)2.37(s,3H),3.76(s,3H),7.26(m,3H),7.50(m,1H), 7.56(s,1H),7.73(d,J=8.4Hz,2H),9.08(s,1H),11.46(m,1H),12.04(m,1H)

[0416] Example 2. Detection of anticancer activity of thiobenzimidazole derivatives

[0417] 2.1 Confirmation of cell viability of thiobenzimidazole derivatives

[0418] In the experiment, human triple-negative breast cancer (TNBC) cell line MDA-MB-231 (cell seeding number: 1 (M231) × 10 4 cells / well (confluence ≥ 25%), and HER2 positive breast cancer (HER2+BC) cell line JIMT-1 (cell seeding number: 1.2 (JIMT) × 10 4 cells / well (confluence ≥ 25%).

[0419] The above cell lines were cultured in DMEM (Dulbecco's modified Eagle's medium) containing 10% fetal bovine serum (FBS), streptomycin-penicillin (100 U / mL) and Fungizone (0.625 μg / mL) in an environment of 5% CO 2 and 37°C.

[0420] In the human breast cancer cell lines MDA-MB-231 and JIMT, the thiobenzimidazole derivatives of Chemical Formulas 1-1, 1-2, 1-3, 1-5, 1-6, 1-7, 1-8, 1-10, 1-11, 1-12, 1-13, 1-14, 1-15, 1-16, 1-17, 1-19, 1-20, 1-21 and 1-22 were treated at different concentrations of 0, 0.5, 1, and 5 μM, respectively, and the cell survival rate was measured using the MTS assay technique after 72 hours. The MTS assay method is as follows: cells are attached to a 96-well plate for 24 hours, then treated with the thiobenzimidazole derivative for 72 hours, followed by color development with MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazole) for 4 hours, and then the absorbance is measured at 490 nm using a Spectramax Plus 384 microplate analyzer.

[0421] The measurement results are as follows Figures 1 to 6 As shown, Figure 1 、 Figure 3 as well as Figure 5 The cell viability of the MDA-MB-231 cell line can be confirmed in the; Figure 2 、 Figure 4 as well as Figure 6 The cell viability of the JIMT-1 cell line can be confirmed.

[0422] As a result, it was confirmed that the thiobenzimidazole derivatives of Chemical Formulas 1-1, 1-2, 1-3, 1-5, 1-6, 1-7, 1-8, 1-10, 1-11, 1-12, 1-13, 1-14, 1-15, 1-16, 1-17, 1-19, 1-20, 1-21 and 1-22 in the human breast cancer cell lines MDA-MB-231 and JIMT-1 mostly inhibited cell survival in a concentration-dependent manner.

[0423] Furthermore, when the HER2-positive breast cancer cell lines SKB R3 and BT474, which are sensitive to trastuzumab, and the HER2-positive breast cancer cell line JIMT-1, which is resistant to trastuzumab, were treated with the thiobenzimidazole derivatives of Chemical Formula 1-3 in a concentration-dependent manner for 72 hours, it was confirmed that all three cell lines showed an inhibitory effect on survival rate. Figure 7 ).

[0424] 2.2 Confirmation of cell cycle arrest and apoptosis of thiobenzimidazole derivatives

[0425] The degree of apoptosis induced by the thiobenzimidazole derivatives of Chemical Formulas 1-3 was measured by DNA content analysis using flow cytometry. For SKBR3, BT474, and JIMT-1 cell lines, a control group (DMSO) and the derivatives of Chemical Formulas 1-3 were treated at concentrations of 0.1, 0.25, and 0.5 μM for 72 hours. The cells were then recovered and fixed with 95% ethanol containing 0.5% Tween-20 for 24 hours and stained with propidium iodide (PI, 50 μg / mL) and RNase (50 μg / mL) for 30 minutes. Thereafter, the degree of apoptosis of the cancer cells was analyzed using flow cytometry.

[0426] Generally speaking, the cell cycle is divided into G1 (cell growth phase) - S (cell replication phase) - G2 / M (cell division phase) according to the DNA content in the cell. When apoptosis is induced, DNA fragmentation occurs, and the DNA content in each cell is significantly lower than that in the G1 phase. The result of this apoptosis appears as a Sub G1 site in the cell cycle. Figure 8 The Sub G1 ratio is shown in numerical terms.

[0427] The results confirmed that the derivatives of Chemical Formulas 1-3 significantly induced apoptosis (Sub-G1 population) and arrested the cell cycle at the G2 / M phase in SKBR3, BT474, and JIMT-1 breast cancer cell lines. Three independent apoptosis experiments were performed and verified by unpaired t-tests (*p<0.01; DMSO control vs NCT-58 or NCT-407). ) significance.

[0428] Furthermore, to confirm the apoptosis-inducing effect of the derivatives of Chemical Formula 1-3, the control group (DMSO) and the derivatives of Chemical Formula 1-3 were treated at concentrations of 0.1, 0.25, and 0.5 μM for 72 hours, and the degree of apoptosis was confirmed by Annexin V / PI staining. As a result, it was confirmed that the treatment with the derivatives of Chemical Formula 1-3 effectively induced early and late apoptosis in a concentration-dependent manner. Figure 9 ).

[0429] Example 3 Preparation of hydrochloride of thiobenzimidazole derivatives

[0430] The following reaction formula 23 shows the synthesis process of the hydrochloride compound 1-23 of the thiobenzimidazole derivative of compound 1-3.

[0431] [Reaction formula 23]

[0432]

[0433] To a suspension of methyl (5-((4-(4-methylpiperazin-1-yl)phenyl)thio)-1H-benzo[d]imidazol-2-yl)carbamate (370 mg, 0.93 mmol) in methanol (75 mL) was injected with hydrogen chloride (HCl) gas every 2 minutes until the pH was less than 1. The reactant was evaporated under reduced pressure to obtain an oily product, to which isopropyl alcohol (37 mL) was added again. The product was heated to dissolve and then cooled to room temperature to obtain a solid suspension. Then, isopropyl ether (35 mL) was added portionwise to the solid suspension, and after stirring for 3 hours, the mixture was filtered. At this time, the mixture was washed with a mixed solvent of IPA and IPE, and dried with hot air at 50° C. for 1 hour and vacuum dried at 40° C. for 2 hours to obtain methyl (5-((4-(4-methylpiperazin-1-yl)phenyl)thio)-1H-benzo[d]imidazol-2-yl)carbamate hydrochloride as an off-white solid (Chemical Formula 1-23, off-white solid, 350 mg, yield 87%).

[0434] 1 H NMR(D2O,400MHz)δ2.93(s,3H),3.08-3.14&3.19-3.25(2m,4H),3.61(d,J=12.5Hz,2H),3.64(d,J=12.5Hz,2H),3.89(s,3H),7.04-7.44(m,7H)

[0435] Example 4. Determination of the anticancer activity of thiobenzimidazole derivatives hydrochloride

[0436] 4.1 Confirmation of cell viability of thiobenzimidazole derivative hydrochloride

[0437] In the HER2-positive breast cancer cell line BT474, which is sensitive to trastuzumab, and the HER2-positive breast cancer cell line JIMT-1, which is resistant to trastuzumab, the thiobenzimidazole derivatives prepared by the above chemical formula 1-3 and their hydrochlorides (compounds 1-23) were treated at the same concentrations (0, 0.1, 0.5, 1, 5, 10 μM) for 72 hours, and the cell viability and IC were measured by MTS assay technology. 50 As a result, in the BT474 cell line, the IC of the derivatives of Chemical Formula 1-3 was 50 was 2.184 μM, while the IC 50The IC values ​​of the derivatives of Chemical Formula 1-3 in JIMT-1 cell line were 0.347 μM, which was more than 6 times lower. 50 The IC value of the hydrochloride salt (Compound 1-23) was 0.448 μM. 50 is 0.228μM, almost half ( Figure 10 ).

[0438] In addition, for triple-negative breast cancer cell lines MDA-MB-231, BT549 and 4T1, the hydrochloride salt of the thiobenzimidazole derivative of the chemical formula 1-3 (compound 1-23) prepared above was treated with different concentrations (0, 0.1, 0.5, 1, 5 and 10 μM) for 72 hours, and the cell viability and IC were measured by MTS assay technology. 50 As a result, it was confirmed that the three cell lines showed low IC values ​​of 0.523μM, 0.559μM, and 0.267μM, respectively. 50 ( Figure 11 ).

[0439] These results indicate that the hydrochloride salt (Compound 1-23) has higher solubility than the derivative of Formula 1-3 and can thus more effectively inhibit cell viability.

[0440] 4.2 Confirmation of Cell Cycle Arrest by Thiobenzimidazole Derivatives Hydrochloride

[0441] HER2-positive breast cancer cell line JIMT-1 and triple-negative breast cancer cell line MDA-MB-231 were treated with different concentrations (0, 0.1, 0.25, 0.5 μM) of the hydrochloride salt of the thiobenzimidazole derivative of Chemical Formula 1-3 (Compound 1-23) for 72 hours, and then subjected to western blot analysis.

[0442] As a result, the hydrochloride of the present invention reduced β-tubulin, which is the main target protein of benzimidazole anthelmintics, and increased p-histone H3 (S10) as a G2 / M cell cycle marker along with the inhibition of tubulin ( Figure 12 ).

[0443] 4.3 Confirmation of the anticancer activity of the hydrochloride salts of thiobenzimidazole derivatives in other cancer cell lines

[0444] In addition to breast cancer cell lines, the blood cancer cell line HL-60, the colorectal cancer cell line HCT116, the non-small cell lung cancer cell lines H1299 and A549, the ovarian cancer cell line SKOV3, the prostate cancer cell line Du145, and the liver cancer cell line HepG2 were treated with different concentrations (0, 0.1, 0.25, 0.5 μM) of the hydrochloride salt of the thiobenzimidazole derivative of Chemical Formula 1-3 (Compound 1-23) and the cell survival rate and IC50 were measured after 72 hours. The results confirmed that the compound also has an excellent effect of inhibiting cell survival in other cancer cells ( Figure 13 as well as Figure 14 ).

[0445] Although this disclosure includes specific examples, it will be apparent to those skilled in the art that various changes in form and detail may be made in these examples without departing from the spirit and scope of the claims and their equivalents. Appropriate results can be obtained if the described techniques are performed in a different order, and / or if the components of the described systems, architectures, devices, or circuits are combined or combined in different configurations, or replaced or substituted with other components or equivalents.

[0446] Accordingly, other embodiments, other examples, and equivalents of the claims are within the scope of the appended claims.

[0447] Industrial Applicability

[0448] The present invention relates to a thiobenzimidazole derivative or a pharmaceutically acceptable salt thereof, and a composition for preventing or treating cancer containing the derivative as an active ingredient. The thiobenzimidazole derivative of the present invention is activated in cancer cells to inhibit tubulin polymerization, and when administered to an individual, can block the cancer cell cycle and induce cell apoptosis to produce cytotoxicity. The thiobenzimidazole derivative can be used to prevent or treat cancer, preferably for preventing or treating triple-negative breast cancer.

Claims

1. A thiobenzimidazole derivative represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof, characterized in that: Chemical formula 1 In the chemical formula 1, R1 is -S-R2, R2 is substituted C3 to C 20 Aryl, the substituted C3 to C 20 Aryl is selected from unsubstituted C3 to C 12 Heterocycloalkyl and unsubstituted C3 to C 12 is substituted by at least one of the group consisting of heteroaryl, wherein the C3 to C 20 The aryl group is phenyl, at least one of the heterocycloalkyl structures is selected from N, O or S, and the heteroaryl group is a monocyclic ring having a N, O or S heteroatom.

2. A thiobenzimidazole derivative or a pharmaceutically acceptable salt thereof, characterized in that: The thiobenzimidazole derivative is selected from any one of the group consisting of the following compounds: [Chemical formula 1-2] [Chemical formula 1-3] [Chemical formula 1-6] [Chemical Formula 1-7] [Chemical Formula 1-8] [Chemical Formula 1-9] [Chemical formula 1-10] [Chemical Formula 1-15] [Chemical Formula 1-16] [Chemical Formula 1-19] [Chemical Formula 1-20] [Chemical Formula 1-21] as well as [Chemical Formula 1-22] 3. The thiobenzimidazole derivative or a pharmaceutically acceptable salt thereof according to claim 2, characterized in that: The thiobenzimidazole derivative is selected from any one of the group consisting of the following compounds: [Chemical formula 1-2] [Chemical formula 1-3] [Chemical formula 1-6] [Chemical Formula 1-7] [Chemical Formula 1-8] [Chemical formula 1-10] [Chemical Formula 1-15] [Chemical Formula 1-16] [Chemical Formula 1-19] [Chemical Formula 1-20] [Chemical Formula 1-21] as well as [Chemical Formula 1-22] 4. The thiobenzimidazole derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, characterized in that: The pharmaceutically acceptable salt of the thiobenzimidazole derivative is any one selected from the group consisting of hydrochloride, bromate, sulfate, phosphate, nitrate, citrate, acetate, lactate, tartrate, maleate, gluconate, succinate, formate, trifluoroacetate, oxalate, fumarate, glutarate, adipate, methanesulfonate, benzenesulfonate, p-toluenesulfonate, camphorsulfonate, sodium salt, potassium salt, lithium salt, calcium salt and magnesium salt.

5. The thiobenzimidazole derivative or a pharmaceutically acceptable salt thereof according to claim 4, characterized in that: The pharmaceutically acceptable salt of the thiobenzimidazole derivative is hydrochloride.

6. A method for preparing the hydrochloride of thiobenzimidazole, characterized in that: The following steps are involved: (1) preparing a suspension of a thiobenzimidazole derivative represented by Chemical Formula 1; [Chemical Formula 1] (2) injecting hydrogen chloride (HCl) into the suspension and evaporating it under reduced pressure; (3) adding isopropyl alcohol to the product of step (2), heating and then cooling; and (4) adding isopropyl ether to the product of step (3), stirring and filtering, In the chemical formula 1, R1 is -S-R2, R2 is substituted C3 to C 20 Aryl, the substituted C3 to C 20 Aryl is selected from unsubstituted C3 to C 12 Heterocycloalkyl and unsubstituted C3 to C 12 is substituted by at least one of the group consisting of heteroaryl, wherein the C3 to C 20 The aryl group is phenyl, at least one of the heterocycloalkyl structures is selected from N, O or S, and the heteroaryl group is a monocyclic ring having a N, O or S heteroatom.

7. A pharmaceutical composition for preventing or treating cancer, characterized in that: The invention comprises the thiobenzimidazole derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5 as an active ingredient.

8. The pharmaceutical composition for preventing or treating cancer according to claim 7, characterized in that The pharmaceutical composition induces cell apoptosis by blocking the cancer cell cycle.

9. The pharmaceutical composition for preventing or treating cancer according to claim 7, characterized in that The cancer is selected from at least one of the group consisting of 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, blood cancer, pancreatic cancer, prostate cancer, testicular cancer, laryngeal cancer, head and neck cancer, thyroid cancer, liver cancer, bladder cancer, osteosarcoma, lymphoma, blood cancer, thymus cancer, urethra cancer and bronchial cancer.

10. The pharmaceutical composition for preventing or treating cancer according to claim 7, characterized in that The cancer is triple-negative breast cancer.

11. Use of a thiobenzimidazole derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5 in the preparation of a medicament for preventing or treating cancer, wherein the cancer is breast cancer, colorectal cancer, lung cancer, ovarian cancer, blood cancer, prostate cancer, or liver cancer.

12. The use according to claim 11, characterized in that The drug induces apoptosis by arresting the cancer cell cycle.

13. The use according to claim 11, characterized in that The cancer is triple-negative breast cancer.

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