WO2023113461A1 - Pharmaceutical composition for preventing or treating cancer - Google Patents

Abstract

The present invention provides a pharmaceutical composition comprising a compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof for prevention or treatment of cancer. Inhibiting the binding or interaction between aurora kinase and MLK3, the compound represented by chemical formula 1 or pharmaceutically acceptable salt thereof according to the present invention has an excellent preventive or therapeutic effect on cancer diseases and is particularly superb in repressing cancer metastasis. Therefore, the pharmaceutical composition comprising a compound represented by chemical formula 1 for prevention and treatment of cancer according to the present invention has excellent effects of lowering cancer cell viability and inhibiting cancer proliferation and migration and thus can be advantageously used as a cancer therapeutic agent.

Description

Pharmaceutical composition for preventing or treating cancer

The present invention relates to a pharmaceutical composition for preventing or treating cancer comprising an inhibitor of the interaction of aurora kinase and mixed lineage kinase 3 (MLK3).

The aurora kinase (AURK) family, including AurkA, B, and C, are cell cycle-regulating serine/threonine kinases whose activity and protein expression are normally associated with important mitosis events, including centrosome maturation, chromosome alignment, chromosome segregation, and cytokinesis. It culminates during mitosis to regulate the process. Aurora kinase includes an N-terminal domain, a kinase domain, and a C-terminal domain, and the kinase domains are AurkA/B, AurkA/C, and AurkB/C. It is highly conserved among Aurora proteins with 71%, 60% and 75% homology, respectively (Willems et al., 2018).

AurkA promotes G2/M transition by promoting centrosome maturation and mitotic spindle assembly. AurkB and AurkC are chromosome-passenger complex proteins, which are important for chromosome binding and chromosome segregation to the kinetochore. The cellular distribution of AurkB is ubiquitous, whereas AurkC expression is mainly restricted to meiotically-active germ cells. In human tumors, all Aurora kinase members play oncogenic roles related to mitotic activity and promote survival and proliferation of cancer cells. In addition, since aurora kinase is overexpressed in various tumor cell lines, the use of aurora kinase small molecule inhibitors can be considered as a potential treatment option for cancer.

On the other hand, protein-protein interactions (PPI) in cells form a complex network that has the term “interactome”. The interactome is responsible for signal transmission, cell proliferation, growth, differentiation and It plays an important role in physiological and pathological processes, including apoptosis, etc. In recent years, some PPI modulators have entered clinical studies, some of which have been approved for clinical use; (Lu et al., 2020), PPI has great potential as an intervention target for new treatment of intractable diseases, and control of PPI is a promising strategy in new drug development. In this regard, one of the major challenges in the development of PPI modulators is to determine their association with diseases among thousands of PPIs and to identify PPIs that can be modulated with drugs.

Under this background, the present inventors have completed the present invention by finding MLK3 (mixed lineage kinase 3) as a protein that interacts with aurora kinase and screening inhibitor candidates that inhibit the interaction between them.

An object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer comprising a compound represented by Formula 1 below or a pharmaceutically acceptable salt thereof.

[Formula 1]

In Formula 1,

R ₁ and R ₂ are the same as or different from each other, and each independently represents a hydrogen atom, a halogen atom, C _1-4 haloalkyl or C _1-6 alkyl;

R ₃ and R ₄ are the same as or different from each other, and are each independently a hydrogen atom, C _1-6 alkyl, substituted or unsubstituted C _3-8 cycloalkyl, substituted or unsubstituted C _3-8 heterocycloalkyl, substituted or C _5-10 heteroaryl containing at least one heteroatom selected from the group consisting of unsubstituted C _5-10 aryl, substituted or unsubstituted N, O and S, or R ₃ and R ₄ are one or more carbon atoms are linked together to form a substituted or unsubstituted C _3-8 heterocycloalkyl or a substituted or unsubstituted C _5-10 heteroaryl, wherein C _3-8 cycloalkyl, C _3-8 hetero At least one H of cycloalkyl, C _5-10 aryl, C _5-10 heteroaryl may be substituted with a halogen atom or C _1-6 alkyl;

p and q are each independently an integer of 0 to 5;

n is any one of the integers of 1-3.

The present inventors completed the present invention through a method for screening cancer drugs according to the presence or absence of inhibition of the interaction between aurora kinase and MLK3 based on a system for confirming the interaction between aurora kinase and MLK3.

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

In the present invention, "Cx-y" means having a carbon number x or more and y or less.

In the present invention, the term "substituted" refers to a moiety having a substituent replacing hydrogen on one or more carbons of the main chain. "Substitution" or "substituted with" is contingent on whether such substitution is permissible on the atom and substituent being substituted, and refers to compounds that are stable by substitution, such as compounds that are not naturally modified by rearrangement, cyclization, elimination, etc. It is defined as including the implicit condition that induces

In the present invention, the term “C _1-6 alkyl” refers to a C _1-6 straight-chain or branched-chain saturated hydrocarbon such as methyl, ethyl, propyl, isopropyl, butyl, pentyl, or hexyl. Preferred alkyl groups contain about 1, 2, 3, 4, 5 or 6 carbon atoms in the chain. Side chain means that one or more lower alkyl groups, such as methyl, ethyl or propyl, are attached to the linear alkyl chain. "Lower alkyl" means a group having from about 1 to about 6 carbon atoms in the chain, which may be straight or branched. “Alkyl” may be unsubstituted or optionally substituted by one or more substituents, which may be the same or different, each substituent being halogen, alkyl, aryl, cycloalkyl, cyano, hydroxy, alkoxy, alkyl thio, amino, carboxy, and the like. Preferably alkyl can be methyl or ethyl. In the present invention, 'methyl (methyl group, methyl group)' is a structure in which three hydrogen atoms are bonded to one carbon atom, and is the smallest alkyl group. This methyl group can exist in three forms: anionic, cationic, and free.

In the present invention, the term “C _1-4 haloalkyl” refers to C _1-4 in which at least one hydrogen of a straight-chain or branched-chain saturated hydrocarbon is substituted with a halogen compound (ie, F, Cl, Br, or _I ). It means a straight-chain or branched-chain saturated hydrocarbon of _-4 . For example, it may be CF ₃ .

In the present invention, the term "C _5-10 aryl" refers to an aromatic hydrocarbon containing 5 to 10 carbon atoms. For example, monocyclic (eg, phenyl); ring systems such as bicyclic (eg, indenyl, naphthalenyl, tetrahydronaphthyl, tetrahydroindenyl). Preferably, aryl has a chemical formula of C ₆ H ₅ and may be a phenyl group in which 6 carbon atoms are arranged in a cyclic ring structure. The phenyl group is a very stable, aromatic hydrocarbon found in many organic compounds. The simplest compound containing a phenyl group is phenol (C ₆ H ₅ OH), which has a structure in which a hydroxy group is bonded. In addition, the hydrogen at the ortho, meta, or para position of the phenyl ring may be substituted with a halogen atom or C _1-4 haloalkyl, preferably F, Cl or CF ₃ substituted. can

In the present invention, the term "C _5-10 heteroaryl" refers to 5 to 10 carbon atoms in which at least one of the ring carbon atoms is replaced with a heteroatom selected from oxygen (O), nitrogen (N) and sulfur (S). refers to an optionally substituted aromatic ring containing, or an aromatic ring (e.g., a bicyclic or tricyclic ring system) fused to more than one ring, such as a heteroaryl ring, aryl ring, heterocyclic ring, or carbocyclic ring; Each may have an optional substituent. Examples include, but are not limited to, isoquinolinyl, pyridinyl, pyrimidinyl, quinolinyl, thiazolyl, furanyl, thiophenyl, indolyl, benzopyridinyl, pyrazolyl, and the like.

In the present invention, the term “C _3-8 cycloalkyl” refers to a saturated hydrocarbon ring containing 3 to 8 carbon atoms, and the saturated hydrocarbon ring is monocyclic and polycyclic, and has a ring structure in which two or more rings share at least one pair of carbon atoms. It means to include all of Cycloalkyl includes, but is not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptanyl, cyclooctanyl, and the like.

In the present invention, "C _3-8 heterocycloalkyl" is a saturated monocyclic and polycyclic heterocycle containing 1 to 4 heteroatoms independently selected from nitrogen (N), oxygen (O) and sulfur (S), or A ring structure in which two or more rings share one or more pairs of carbon atoms. Heterocycloalkyl includes oxiranyl, oxetanyl, mophorinyl, thietanyl, pyrrolidinyl, piperidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyranyl, tetrahydrothiopyranyl, and the like. And, preferably, it may be piperidinyl, but is not limited thereto.

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

A pharmaceutical composition for preventing or treating cancer comprising the compound of Formula 1 or a pharmaceutically acceptable salt thereof

In one aspect of the present invention, a pharmaceutical composition for preventing or treating cancer comprising a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof is provided.

[Formula 1]

In Formula 1,

R ₁ and R ₂ are the same as or different from each other, and each independently represents a hydrogen atom, a halogen atom, C _1-4 haloalkyl or C _1-6 alkyl;

R ₃ and R ₄ are the same as or different from each other, and are each independently a hydrogen atom, C _1-6 alkyl, substituted or unsubstituted C _3-8 cycloalkyl, substituted or unsubstituted C _3-8 heterocycloalkyl, substituted or C _5-10 heteroaryl containing at least one heteroatom selected from the group consisting of unsubstituted C _5-10 aryl, substituted or unsubstituted N, O and S, or R ₃ and R ₄ are one or more carbon atoms are linked together to form a substituted or unsubstituted C _3-8 heterocycloalkyl or a substituted or unsubstituted C _5-10 heteroaryl, wherein C _3-8 cycloalkyl, C _3-8 hetero at least one H of cycloalkyl, C _5-10 aryl or C _5-10 heteroaryl may each independently be substituted with a halogen atom or C _1-6 alkyl;

p and q are each independently an integer of 0 to 5;

n is any one of the integers of 1-3.

In one embodiment of the present invention, the compound represented by Formula 1, or a pharmaceutically acceptable salt thereof, may be preferably the following.

In Formula 1 of the present invention, p may be any one of an integer of 0, 1 or 2. Here, when p is 0, it can be seen that a hydrogen atom is substituted.

In Formula 1 of the present invention, q may be any one of 0, 1 or 2 integers. When p is 0, it can be seen that a hydrogen atom is substituted.

In Formula 1 of the present invention, R ₁ may be a halogen atom, C _1-4 haloalkyl or C _1-6 alkyl, and R ₁ may be a halogen atom, C _1-4 haloalkyl or C _1-6 alkyl substituted. If not, hydrogen atoms are generally substituted. More specifically, R ₁ may be F, Cl, Br, I, CF ₃ , methyl, ethyl or propyl, preferably R ₁ may be F, Cl or CF ₃ .

In Formula 1 of the present invention, q is 0, which means that hydrogen atoms are substituted on all five carbon atoms of the phenyl group.

In addition, the compound represented by Formula 1 may be a compound represented by Formula 1-1 below or a pharmaceutically acceptable salt thereof.

[Formula 1-1]

In Formula 1-1,

R _1a , R _1b and R _1c are the same as or different from each other, and each independently represents a hydrogen atom, a halogen atom or a C _1-4 haloalkyl;

R ₃ and R ₄ are the same as or different from each other, and are each independently a hydrogen atom, C _1-6 alkyl, substituted or unsubstituted C _3-8 cycloalkyl, substituted or unsubstituted C _3-8 heterocycloalkyl, substituted or C _5-10 heteroaryl containing at least one heteroatom selected from the group consisting of unsubstituted C _5-10 aryl, substituted or unsubstituted N, O and S, or R ₃ and R ₄ are one or more carbon atoms are linked together to form a substituted or unsubstituted C _3-8 heterocycloalkyl or a substituted or unsubstituted C _5-10 heteroaryl, wherein C _3-8 cycloalkyl, C _3-8 hetero One or more H of cycloalkyl, C _5-10 aryl or C _5-10 heteroaryl may each independently be substituted with a halogen atom or C _1-6 alkyl.

In Formula 1-1, R _1a may be a hydrogen atom, a halogen atom, or C _1-4 haloalkyl, R _1b may be a hydrogen atom or a halogen atom, and R _1c may be a hydrogen atom or a halogen atom.

More specifically, when R _1a is C _1-4 haloalkyl, R _1b is a hydrogen atom, R _1c is a hydrogen atom, and preferably R _1a is CF ₃ .

Further, when R _1a is a halogen atom, R _1b is a hydrogen atom or a halogen atom, R _1c is a hydrogen atom, and preferably R _1b is a hydrogen atom, F or Cl.

Further, when R _1a is a hydrogen atom, R _1b is a halogen atom, R _1c is a halogen atom, and preferably, R _1b and R _1c are each independently Cl.

In Formula 1 or Formula 1-1, when any one of R ₃ and R ₄ is a hydrogen atom, the other one may be a substituted or unsubstituted C _5-10 aryl.

Preferably, the substituted or unsubstituted C _5-10 aryl may be a substituted or unsubstituted phenyl, more preferably, one or more H of the phenyl may be each independently substituted with a halogen atom, and the halogen may be F can

Also, in one embodiment, R ₃ and R ₄ are linked together with one or more carbon atoms to include a substituted or unsubstituted N atom, C _3-8 heterocycloalkyl containing a substituted or unsubstituted N atom, or C _5-10 heteroaryl may be formed. When R ₃ and R ₄ are linked together with one or more carbon atoms to form a ring containing a nitrogen atom (N), C _3-8 heterocycloalkyl or C _5-10 heteroaryl containing a nitrogen atom (N) is formed. can form

Preferably in Formula 1 or Formula 1-1,

is formed by connecting R ₃ and R ₄ together with one or more carbon atoms

or

and the above

or

is unsubstituted, or at least one H may be each independently substituted with a halogen atom, C _1-4 haloalkyl or C _1-6 alkyl, preferably a halogen atom or C _1-6 alkyl.

Specifically, the above

or

is unsubstituted, or at least one H may be independently substituted with methyl, F, Cl, Br, I or CF ₃ , more preferably the above

may be substituted with methyl.

According to an embodiment of the present invention, the compound of Formula 1 may be any one selected from the group consisting of the following compounds:

N-[2-(4-methyl-1-piperidinyl)-2-oxoethyl]-N-[3-(trifluoromethyl)phenyl]benzenesulfonamide;

N~1~-phenyl-N~2~-(phenylsulfonyl)-N~2~-[3-trifluoromethyl)phenyl]glycinamide;

N~1~-(4-fluorophenyl)-N~2~-(phenylsulfonyl)-N~2~-[3-(trifluoromethyl)phenyl]glycinamide;

N-(3,4-dichlorophenyl)-N-[2-(4-methyl-1-piperidinyl)-2-oxoethyl]benzenesulfonamide;

N-(3-chloro-4-fluorophenyl)-N-[2-oxo-2-(1-piperidinyl)ethyl] benzenesulfonamide; and

N-(3,4-dichlorophenyl)-N-[2-(3,4-dihydro-2(1H)-isoquinolinyl)-2-oxoethyl]benzenesulfonamide.

Preferably, the compound of Formula 1 may be N-[2-(4-methyl-1-piperidinyl)-2-oxoethyl]-N-[3-(trifluoromethyl)phenyl]benzenesulfonamide. there is.

According to an embodiment of the present invention, the compound of Formula 1 may be any one selected from the group consisting of the following compounds 1 to 6:

In one embodiment of the present invention, a compound that inhibits the interaction between aurora kinase (AURK) and MLK3 was screened through screening of a compound library by the screening system developed by the inventors of the present invention by interaction detection. As a result, it was confirmed that the compound represented by Formula 1, preferably the compounds 1 to 6, more preferably the compound 1 inhibits the interaction or binding of aurora kinase (AURK) and MLK3. It was confirmed in the examples of the present invention that the compounds 1 to 6 had the effect of significantly lowering the binding affinity between MLK3 and AURKA, the binding affinity between MLK3 and AURKB, and the binding affinity between MLK3 and AURKC (FIG. 8). It was confirmed that the compound represented by Chemical Formula 1 of the present invention can be used as an inhibitor for inhibiting the interaction and binding between aurora kinase and MLK3.

The term "interaction" used in the present invention means direct binding of two proteins, Aurora Kinase (AURK) and MLK3.

As used herein, the term "interaction inhibition" includes inhibition, reduction, or elimination of the interaction between aurora kinase (AURK) and MLK3. Interaction inhibition may inhibit the mutual interaction of aurora kinase (AURK) and MLK3, but also includes inhibition of mutual binding by reducing the activity of aurora kinase (AURK) or MLK3.

In one embodiment, when the compounds 1 to 6 were treated by expressing aurora kinase (AURK) and MLK3, interactions between proteins were found to decrease (FIGS. 6 to 8). In particular, it was found that the effect of inhibiting cancer proliferation and metastasis was excellent.

Accordingly, it was confirmed that the compound represented by Chemical Formula 1 of the present invention induces the death of cancer cells as an inhibitor of the interaction between aurora kinase (AURK) and MLK3, and exhibits anticancer effects by inhibiting metastasis and proliferation thereof.

The term "anti-cancer" used in the present invention refers to the action of suppressing the proliferation or killing of cancer cells, and refers to both prevention and treatment of cancer. “Inhibiting or preventing the growth of cancer” is a concept that includes reducing cancer growth and cancer metastasis compared to treatment or no treatment.

As used herein, the term “prevention” refers to any activity that inhibits cancer formation or delays the onset of cancer by administering a composition.

In the present invention, "treatment" refers to all activities in which the symptoms of the disease are improved or beneficially changed by administration of the composition.

In the present invention, the cancer is breast cancer, brain cancer, nerve cancer, colorectal cancer, lung cancer, small cell lung cancer, stomach cancer, liver cancer, blood cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, skin or intraocular melanoma, cervical cancer, ovarian cancer , rectal cancer, perianal cancer, colon cancer, breast cancer, fallopian tube carcinoma, endometrial carcinoma, cervical cancer, vaginal cancer, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethra cancer, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, renal cell carcinoma, renal pelvic carcinoma, CNS tumor, primary CNS lymphoma, spinal cord tumor, brain stem glioma, and pituitary adenoma. there is.

Preferably, the cancer may be metastatic carcinoma.

Preferably, the cancer may be breast cancer, more preferably triple-negative breast cancer or HER2-positive breast cancer, and even more preferably triple-negative breast cancer.

In one embodiment of the present invention, the compound represented by Formula 1 of the present invention (eg, Compound 1) has a therapeutic effect on breast cancer (cancer cell proliferation inhibition and metastasis inhibition effect) through a triple negative breast cancer (TNBC) mouse animal model. It was confirmed that it was significantly superior (Figs. 12 to 17).

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

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

The pharmaceutically acceptable carrier includes those commonly used in the art, such as lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like. In addition, the pharmaceutical composition of the present invention may include diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, and other pharmaceutically acceptable additives, but is not limited thereto.

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

When the pharmaceutical composition of the present invention is formulated in liquid form for oral use, it includes suspensions, internal solutions, emulsions, syrups, etc., and diluents such as water and liquid paraffin, wetting agents, sweeteners, aromatics, preservatives, etc., but are not limited thereto.

When the pharmaceutical composition of the present invention is formulated for parenteral use, it includes a sterilized aqueous solution, a non-aqueous solvent, a suspension, an emulsion, a lyophilized preparation, and a suppository, and the non-aqueous solvent and suspension include propylene glycol, polyethylene glycol, and olive vegetable oils such as oils, injectable esters such as ethyl oleate, and the like, but are not limited thereto. As a base material for suppositories, witepsol, macrogol, tween 61, cacao paper, laurin paper, glycerogelatin, etc. may be used, but are not limited thereto.

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

The pharmaceutical composition of the present invention can be administered to mammals such as rats, mice, livestock, and humans through various routes, for example, oral administration, intrathecal, inner ear, intraperitoneal or intravenous, intramuscular, subcutaneous, intrauterine, sublingual or cerebrovascular It may be administered by intravenous injection, but is not limited thereto.

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

A method for preventing or treating cancer comprising administering the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof to a subject in need thereof, and uses of the compound of Formula 1

Another aspect of the present invention provides a method for preventing or treating cancer comprising administering the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof to a subject in need thereof.

In the present invention, the term "compound represented by Formula 1" or "cancer" is as described above.

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

The term "administration" of the present invention means introducing a predetermined substance into a patient by an appropriate method, and the administration route of the composition may be administered through any general route as long as it can reach the target tissue. In addition, the pharmaceutical composition of the present invention may be administered by any device capable of transporting an active substance to a target tissue. For example, oral administration, intrathecal administration, intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, topical administration, intranasal administration, intrapulmonary administration, intrarectal administration, inner ear administration, intrauterine intrauterine administration, It may be administered by sublingual administration or intracerebrovascular injection, but is not limited thereto. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and preparations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories.

Administration of the pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. Considering all of the above factors, it can be administered in an amount that can obtain the maximum effect with the minimum amount without side effects, which can be easily determined by those skilled in the art.

Another aspect of the present invention provides the use of the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof of the present invention for preparing a medicament for preventing or treating cancer.

Another aspect of the present invention provides a composition comprising the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof for use in preventing or treating cancer.

The present invention provides a pharmaceutical composition for preventing and treating cancer comprising a compound that inhibits the interaction between aurora kinase and MLK3. Specifically, the compound represented by Formula 1 of the present invention, or a pharmaceutically acceptable salt thereof, inhibits the binding or interaction between aurora kinase and MLK3, and thus has excellent preventive or therapeutic effects on various cancer types, In particular, the effect of inhibiting cancer metastasis is excellent. Therefore, the pharmaceutical composition for preventing and treating cancer comprising the compound represented by Formula 1 of the present invention is useful as a cancer therapeutic agent in that it has excellent effects of lowering cancer cell viability and inhibiting cancer proliferation and migration. can

Figure 1 shows the results confirming the transformation of NIH3T3 normal fibroblasts when treated with control (WT), MLK3, aurora kinase family (AURKA, AURKB, AUKC), aurora kinase family/MLK3.

Figure 2 shows the results of confirming and quantifying the transformation of NIH3T3 normal fibroblasts when treated with control (WT), MLK3, aurora kinase family (AURKA, AURKB, AUKC), aurora kinase family/MLK3. 2, WT (WT), M (MLK3), A (AURKA), B (AURKB), C (AURKC), A / M (AURKA / MLK3), B / M (AURKB / MLK3), C / M ( AURKC/MLK3).

Figure 3 shows the results of Western blot analysis of the expression levels of MLK3 and aurora kinase family (AURKA, AURKB, AUKC) proteins in the invasive breast cancer cell line MDA-MB-231.

Figure 4 shows the results of analyzing the effect of the interaction of MLK3 and the aurora kinase family (AURKA, AURKB, AUKC) in the invasive breast cancer cell line MDA-MB-231.

Figure 5 shows the results of quantifying the number of colonies upon treatment with MLK3 and Aurora kinase family (AURKA, AURKB, AUKC) in the invasive breast cancer cell line MDA-MB-231. 5, G (GFP), R (RFP), G / R (GFP / RFP), G-A (G-AURKA), G-B (G-AURKB), G-C (G-AURKC), R-M (R-MLK3), G-A/R-M (G-AURKA/R-MLK3), G-B/R-M (G-AURKB/R-MLK3), and G-C/R-M (G-AURKC/R-MLK3).

Figure 6 shows the results of analyzing the luciferase activity (luciferase) to confirm the effect of the compound of formula 1 of the present invention on the interaction and binding of aurora kinase and MLK3.

Figure 7 shows the results of analyzing the luciferase activity (luciferase) to confirm that Compound 1 of the present invention inhibits cell migration.

8 shows the effect of inhibiting the binding affinity of MLK3 and aurora kinase (AURKA, AURKB, AURKC) by compounds 2 to 6 of the present invention.

Figure 9 shows the results of measuring the anticancer efficacy of Compound 1 in MDA-MB-231.

10 is a result of colony formation analysis of cells to confirm that the compound of formula 1 (inhibitor) of the present invention exerts an anticancer effect by targeting the interaction between aurora kinase and MLK3 protein in MDA-MB-231 cells. indicates

Figure 11 shows the results of migration analysis in MDA-MB-231 breast cancer cells to confirm that the compound of Formula 1 (Inhibitor) of the present invention exerts an anticancer effect by targeting the interaction between Aurora kinase and MLK3 protein. .

Figure 12 shows the results of measuring the body weight of mice when the mouse TNBC model was treated with Compound 1.

Figure 13 shows the results of analyzing the tumor volume when the mouse TNBC model was treated with Compound 1.

Figure 14 shows the fluorescence imaging results of mouse tumors when the mouse TNBC model was treated with Compound 1.

15 shows the result of measuring the total flux of the labeled tumor distribution when the mouse TNBC model was treated with Compound 1.

Figure 16 shows tumor tissue excised from the mouse model when compound 1 was treated with the mouse TNBC model.

17 shows the results of measuring the weight of tumors excised from the mouse model when the mouse TNBC model was treated with Compound 1.

18 is a result of analyzing cancer cell viability by time and concentration for analyzing the killing ability of Compound 1 of the present invention on various cancer cells.

19 is a result of analyzing cancer cell viability by time and concentration for analyzing the killing ability of Compound 2 of the present invention on various cancer cells.

20 is a result of analyzing cancer cell viability by time and concentration for analyzing the killing ability of Compound 3 of the present invention on various cancer cells.

21 is a result of analyzing the killing efficacy of Compound 4 of the present invention on various cancer cells.

22 is a result of analyzing cancer cell viability by time and concentration for analyzing the killing ability of Compound 5 of the present invention on various cancer cells.

23 is a result of analyzing cancer cell viability by time and concentration for analyzing the killing ability of Compound 6 of the present invention on various cancer cells.

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

<Example>

analysis method

1. Cell culture and treatment

The human breast cancer cell lines MDA-MB-231, MCF7, and MDA-MB-468, the human colorectal cancer cell line HCT116(p53+), the human brain tumor cell line U87MG, the human neural cancer cell line SH-SY5Y, and the human cervical cancer cell line Hela are derived from the Korea Cell Line Bank (KCLB, Seoul). , South Korea), the normal mammary epithelial cell line MCF10A was obtained from the Experimental Animal Resource Center (Korea Research Institute of Bioscience and Biotechnology). HEK293T cells were obtained from the Korean Cell Line Bank (Seoul, Korea). HEK293T, MCF7, MDA-MB-231, U87MG,SH-SY5Y and MDA-MB-231 cells were grown on HyClone (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% FBS in a 37°C, 5% CO ₂ incubator. of DMEM (Dulbecco's modified Eagle's medium). MCF10A cells were cultured in DMEM/F12 (1:1) from HyClone (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% FBS in a 37°C, 5% CO ₂ incubator. Hela cells were cultured in Minimum Essential Medium (MEM) from HyClone (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% FBS in a 37°C, 5% CO ₂ incubator. HCT116(p53+) cells were cultured in RPMI-1640 from HyClone (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% FBS in a 37°C, 5% CO ₂ incubator. Only cells up to passage 15 were used in the experiments.

Compounds 1 to 6 of the present invention, which are aurora kinase/MLK3 interaction inhibitors, were provided by ChemBridge Corporation (San Diego, USA). Stock solutions thereof were prepared in dimethylsulfoxide (DMSO) and added directly to the culture medium.

Control cells were treated only with DMSO, and the final DMSO concentration was always <0.2%.

2. Cell Imaging for PPI Analysis

The cDNAs of 520 human kinases were amplified by PCR and introduced into the mammalian expression plasmid pEGFP-C3 (Han et al., 2017). For PPI screening, cells were seeded in 96-well plates and HEK293T cells were composed of C-terminal fusion construct PKCδ/monomer red fluorescent protein (PKCδ/mRFP)-bait (AurkA, AurkB, and AurkC) and eGFP-prey protein (including MLK3, 597 kinase). Fluorescent cell images were obtained using a confocal laser scanning fluorescence microscope (LSM710, Carl Zeiss) equipped with a 10X objective.

A positive reaction confirmed that the cells had grown into 6-well culture plates on glass coverslips (Knittel Glaser, Braunschweig, Germany) to 50-70% confluence. Transient co-eukaryotic transformation of PKCδ/mRFP-tagged bait (empty, AurkA, AurkB, and AurkC) and eGFP-tagged prey (MLK3) protein-coding plasmid pairs was performed using turbofect according to the manufacturer's standard protocol. did For live cell imaging, transiently co-transfected cells on glass coverslips were mounted in a live cell chamber connected to a temperature and CO ₂ controller (Heating & Cooling system/CU-501 and Gas Mixer/FC-5N, LCI). . After washing the cells with medium (DMEM without phenol red, 10% FBS, 37° C.), 1 ml of medium was added. Consecutive images of the same cells were taken at 2 min intervals using a laser scanning confocal microscope (LSM 710, Carl Zeiss, Germany) equipped with a C-Apochromat 40X/1.2 water immersion lens (488 nm argon laser/505-550). minutes (488 nm argon laser/505-550 nm detection range for eGFP, 561 nm solid-state laser/586-662 nm detection range for mRFP). During imaging, PMA (total concentration of PMA in the medium is 500 nM) was added to the chamber.

3. Mammalian 2 Hybrid (M2H) Assay

The CheckMate M2H system (Promega, Madison, WI, USA) contains three expression vectors, termed pG5luc, pBIND and pACT, which contain the yeast GAL4 DNA-binding domain upstream of the multiple cloning region and eukaryotic transformation control. expresses the SV40 control Renilla luciferase for The pACT vector contains the herpes simplex VP16 activation domain upstream of the multicopy region.

The genetic information encoding the interacting proteins (AurkC, AurkA, AurkB, MLK3-FL, MLK3-KD and MLK3-CRIB) were then cloned into pBIND and pACT vectors to fusion proteins with the DNA binding domains of the activation domains of GAL4 and VP16. was created. GAL4 and VP16 fusions were eukaryotic transformed in HEK293T cells.

Relative luciferase activity to pG5-luc was determined by normalizing firefly luciferase activity to renilla luciferase activity, and luciferase activity was determined using the Dual-Glo Luciferase Assay System kit (Promega, Madison, WI, USA).

After 24 hours of eukaryotic transformation, the cells were washed once consecutively with phosphate buffered saline (PBS). After adding 200 μl of lysis buffer, cells were harvested and centrifuged (4° C., 13,000 rpm, 5 min). The measurement was performed by incubating the cell extract for 10 minutes within 2 hours after diluting the cell extract 1:1 using the Dual-Glo luciferase reagent, and all analyzes were performed in triplicate.

4. Immunoblot (IB) analysis

All cell extracts were harvested in 1X RIPA buffer (2X, 1 M tris pH 7.5, 4 M NaCl, 200 mM EDTA, 10% NP-40) and samples were centrifuged for 30 min at 13,000 rpm at 4 °C. Afterwards, the samples were boiled in sample loading buffer containing SDS (Invitrogen, Carlsbad, CA, USA), and the same amount of samples was separated on a 10% SDS-PAGE gel and then transferred to a PVDF membrane (Bio-Rad, Hercules, CA, USA). moved to

Membranes were blocked and incubated overnight at 4 °C with the indicated primary antibody, followed by incubation with horseradish peroxidase (HRP) conjugated secondary antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA). Proteins were visualized using an enhanced chemiluminescence (ECL) detection system (GE Healthcare Bio-Sciences Corp., Piscataway, NJ, USA), and all immunoblot analysis was performed using a ChemiDac™ XRS+ imaging system (Bio-Rad, Hercules, CA, USA). USA) was used. The intensity of each protein band was normalized to that of β-actin to generate a relative intensity.

5. Foci assay

1000 cells were seeded in a 6-well plate and cultured for 10 days to form colonies. After incubation, cells were washed with PBS and stained with 0.1% crystal violet (Amersco, Solon, OH, USA) in 50% methanol for counting.

6. Soft agar assay

For the soft agar assay, 1.2% agarose (Promega, Madison, WI) dissolved in medium was plated at the bottom of each well. Cells were trypsinized to generate a single cell suspension. Single cell suspensions (1000 cells per well) were mixed with 0.6% agarose and seeded onto bottom agar, and all assays were performed in triplicate. Cells were incubated at 37° C. for 6 weeks to form colonies, and images were captured using a Zeiss inverted wide-field microscope with a Canon G12 camera.

7. Cancer cell apoptosis assay

For the cytotoxicity of the compounds used in the examples of the present invention, a commonly used CCK8 kit (Dojindo Molecular Technologies) was used, and all conditions were performed according to the manufacturer's instructions. More specifically, in the case of the compound of Formula 1, the cells were cultured on a 96-plate for one day, treated with 1, 10, and 50 μM of the compound, cultured for 24, 48, and 72 hours, and then 10 μl of CCK8 solution was added and 450 Absorbance was measured in nm. In the case of the compounds of formulas 1 to 6, the cells were cultured on a 96-plate for one day, treated with 1, 10, and 50 μM compounds, cultured for 24 and 72 hours, and then 10 μl of CCK8 solution was added and absorbance at 450 nm after one hour was measured.

8. Statistical Analysis

Results are presented as mean ± standard deviation of a representative experiment (n = 3) and error bars represent standard deviation. All statistical analyzes were performed using Student's t-test or Mann-Whitney U-test if data were not normally distributed. A difference was considered statistically significant if the null hypothesis could be rejected with 95% or greater confidence (p < 0.05).

Example 1. Analysis of the influence of the interaction between Aurora kinase and MLK3 ( In vitro )

Example 1-1. Effects of aurora kinase/MLK3 binding on NIH3T3 normal fibroblasts

It was confirmed through analysis that the Aurora kinase/MLK3 interaction promotes malignant cell transformation of normal NIH3T3 fibroblasts, and the results are shown in FIGS. 1 and 2 . Referring to Figures 1 and 2, it was confirmed that overexpression of aurora kinase induced normal cell transformation. In addition, MLK3 and Aurora kinase double eukaryotic transformation was elevated in cells overexpressing the single protein, and the interaction of Aurora kinases (AURKA, AURKB, AURKC) with MLK3 promoted malignant cell transformation of NIH3T3 normal fibroblasts.

Example 1-2. Effects of aurora kinase/MLK3 binding on breast cancer cells

Figure 3 confirms the expression of the invasive breast cancer cell line MDA-MB231, the expression of the aurora kinase family (AURKA, AURKB, AURKC) and MLK3 was found to be significant, and the aurora kinase family and MLK3 expression are involved in metastatic breast cancer cells confirmed that

In addition, MDA-MB-231 cells expressing Aurora kinase and MLK3 were constructed using a lentivirus system and soft agar assay was performed. The results are shown in FIGS. 4 and 5 .

Referring to FIGS. 4 and 5 , when Aurora kinase was overexpressed, the growth of MDA-MB-231 cells was rapidly increased, and colonization was particularly prominent in the group treated with Aurora kinase/MLL3. That is, it was confirmed that the binding of Aurora kinase/MLL3 promotes the growth of MDA-MB-231 and greatly increases the level of metastasis.

Example 2. Confirmation of the binding inhibitory effect of the compound of Formula 1 of the present invention on Aurora kinase and MLK3 ( In vitro )

It was confirmed that the compound represented by Formula 1 of the present invention inhibits the protein-protein interaction (PPI) of Aurora kinase and MLK3 through luciferase activity analysis and binding affinity analysis, and the results are shown in FIGS. 6 to 8 had shown

Referring to Figures 6 and 7, compared to the group not treated with the compound (Vehicle), it was found that the luciferase activity was significantly lowered when the compound 1 of the present invention was treated (FIG. 6), Bind-aurora It was shown that luciferase activity of transient eukaryotic transformation was inhibited by kinase and Act-MLK3-KD (FIG. 7).

In addition, referring to FIG. 8 and Table 1, when compounds 2 to 6 were treated, the binding affinity between Aurora kinase and MLK3 was found to be lowered to 80% or less (Table 1).

compound	MLK3/aurora kinase binding affinity (%)
	AURKA	AURKB		AURKC
3	70.8±6.5	55.3±2.2	56.3±1.1
4	55.8±3.2	57.4±4.4	57.8±4.2
6	73.2±4.0	61.9±3.0	67.6±4.0
2	67.3±4.5	59.6±2.9	77.0±4.7
5	53.0±4.6	61.7±5.9	64.8±3.6

Example 3. Cytotoxicity analysis of the compound of Formula 1 of the present invention on breast cancer cells ( In vitro )

In order to confirm whether the antiproliferative effect of Compound 1 of the present invention is specific to cancer cells, normal cells and cancer cells were treated with the drug, and proliferation was measured using a CCK8 kit, and the results are shown in FIG. 9 .

Referring to FIG. 9, MDA-MB-231 cells showed a decrease in proliferation when treated with 25 μM of Compound 1 for 24 hours. Increased killing of the invasive breast cancer cell line MDA-MB-231 was shown. In contrast, MCF10A, which is a normal breast epithelial cell, did not show a significant decrease in proliferation under the same conditions. Therefore, it was confirmed that the triple-negative breast cancer cells were specifically sensitive to the antiproliferative effect of Compound 1.

In addition, metastasis and colony formation assays were performed in MDA-MB-231 breast cancer cells to confirm that compound 1 of the present invention exerts anticancer effects by targeting the interaction between aurora kinase and MLK3 protein, and the results are shown in FIG. 10 and 11.

10 and 11, Compound 1 significantly reduced the colony formation of MDA-MB-231 cells in a concentration-dependent manner (Fig. 10), and when MDA-MB-231 cells were treated with Compound 1, the cell migration rate increased. It decreased in a concentration dependent manner (FIG. 11).

Example 4. Breast cancer treatment effect of compound 1 of the present invention in animal models ( in vivo )

MDA-MB-231-Red-Fluc cells were generated in a lentivirus system encoding red-shifted firefly luciferase. To establish MDA-MB-231-Red-Fluc-GFP cells, Red-Fluc-GFP was transduced into MDA-MB-231 cells by a lentiviral vector. Transduced MDA-MB-231 cells were selected by puromycin for the next 1 week. To establish a mouse TNBC model, 2×10 ⁶ MDA-MB-231-Red-Fluc cells in 100 μL of phosphate buffered saline (PBS) were subcutaneously injected into the right flank of 6-week-old male SCID mice (Nara Biotech, Gyeonggi-do). On days 2, 4, 7 and 9 after tumor cell inoculation, mice were intratumorally injected with Compound 1 suspended in 100 μL of PBS. PBS was administered to control mice for negative and positive antitumor effects. After tumor inoculation, mice were intraperitoneally injected with 150 mg/kg of D-luciferin (XenoLight® D-Luciferin-K+ substrate, Perkin Elmer) at weekly intervals from day 1, and tumors were examined after anesthesia with 1-3% isoflurane. Tumor development was investigated by biofluorescence imaging (BLI) using an in vivo imaging system (IVIS Spectrum, Perkin Elmer, Hopkinton, MA, USA) with Living Image software (Perkin-Elmer). All animal testing procedures followed the guidelines of the Korea Basic Science Institute Animal Laboratory Steering Committee, and the results are shown in FIGS. 12 to 17.

Referring to Figures 12 and 13, compared to the control group MOCK, there was little change in body weight in the compound 1-administered group (Inhibitor), but the tumor volume was significantly reduced.

In addition, as can be seen in FIG. 14 showing mouse biofluorescence imaging results and FIG. 15 quantifying the total flux of the labeled tumor distribution, the tumor volume was significantly reduced compared to the control group (Vehicle).

In addition, as a result of measuring the tumor tissue excised from the mouse model (FIG. 16) and its weight (FIG. 17), it was found that the size of the tumor was significantly reduced by administering Compound 1, an inhibitor of the interaction between MLK3 and aurora kinase of the present invention. appear.

Example 5. Analysis of cancer cell killing ability of the compound according to the present invention

In order to analyze the killing efficacy of the compounds of Formulas 1 to 6 of the present invention on various cancer cells, the concentration and time-dependent cancer cell viability were analyzed, and the results are shown in FIGS. 18 to 23.

Referring to FIG. 18, the composition containing Compound 1 showed effects at different concentrations and over time in breast cancer cell lines MCF7, MDA-MB231, and MDA-MB468, confirming the ability to kill cancer cells, and other cell lines, U87MG (brain cancer cells), SH- SY5Y (neural cancer cells), HeLa (cervical cancer), and HCT116_p53+ (colon cancer) also showed significant effects by concentration and time, confirming their ability to kill cancer cells.

Referring to FIG. 19, the composition containing compound 2 is MCF7, MDA-MB231, MDA-MB468 (breast cancer cell line), U87MG (brain cancer cell), SH-SY5Y (neural cancer cell), HeLa (cervical cancer), HCT116_p53+ (colon cancer cell) The cancer cell killing effect on cancer) was confirmed.

Referring to FIG. 20, the composition containing Compound 3 kills cancer cells against MCF7, MDA-MB231, MDA-MB468 (breast cancer cell line), SH-SY5Y (neural cancer cell), HeLa (cervical cancer), and HCT116_p53+ (colorectal cancer). effect was confirmed.

Referring to Figure 21, the composition containing compound 4 is MCF7, MDA-MB231, MDA-MB468 (breast cancer cell line), U87MG (brain cancer cell), SH-SY5Y (neural cancer cell), HeLa (cervical cancer), HCT116_p53+ (colon cancer cell) The cancer cell killing effect on cancer) was confirmed.

Referring to FIG. 22, the composition containing compound 5 is MCF7, MDA-MB231, MDA-MB468 (breast cancer cell line), U87MG (brain cancer cell), SH-SY5Y (neural cancer cell), HeLa (cervical cancer), HCT116_p53+ (colon cancer cell) The cancer cell killing effect on cancer) was confirmed.

Referring to FIG. 23, the composition containing compound 6 kills cancer cells against MDA-MB231, MDA-MB468 (triple-negative breast cancer cell line), U87MG (brain cancer cells), SH-SY5Y (neural cancer cells), and HeLa (cervical cancer). effect was confirmed.

Through the above results, it was further confirmed that the compounds according to the present invention exhibit excellent anticancer efficacy against various types of carcinoma. Through this, it was confirmed that the compound according to the present invention has an excellent effect on inhibiting the growth and metastasis of various cancers, and is highly likely to be used as a cancer treatment.

Claims (20)

1. A pharmaceutical composition for preventing or treating cancer comprising a compound represented by Formula 1 below, or a pharmaceutically acceptable salt thereof:

   [Formula 1]

   

   In Formula 1,

   R ₁ and R ₂ are the same as or different from each other, and each independently represents a hydrogen atom, a halogen atom, C _1-4 haloalkyl or C _1-6 alkyl;

   R ₃ and R ₄ are the same as or different from each other, and are each independently a hydrogen atom, C _1-6 alkyl, substituted or unsubstituted C _3-8 cycloalkyl, substituted or unsubstituted C _3-8 heterocycloalkyl, substituted or C _5-10 heteroaryl containing at least one heteroatom selected from the group consisting of unsubstituted C _5-10 aryl, substituted or unsubstituted N, O and S, or R ₃ and R ₄ are one or more carbon atoms are linked together to form a substituted or unsubstituted C _3-8 heterocycloalkyl or a substituted or unsubstituted C _5-10 heteroaryl, wherein C _3-8 cycloalkyl, C _3-8 hetero At least one H of cycloalkyl, C _5-10 aryl, or C _5-10 heteroaryl may each independently be substituted with a halogen atom or C _1-6 alkyl;

   p and q are each independently an integer of 0 to 5;

   n is any one of the integers of 1-3.
2. The method of claim 1, wherein in Formula 1, p is any one of integers from 1 to 2;

   R ₁ is a halogen atom, C _1-4 haloalkyl or C _1-6 alkyl, the pharmaceutical composition.
3. The pharmaceutical composition according to claim 1, wherein the compound represented by Formula 1 is a compound represented by Formula 1-1, or a pharmaceutically acceptable salt thereof:

   [Formula 1-1]

   

   In Formula 1-1,

   R _1a , R _1b and R _1c are the same as or different from each other, and each independently represents a hydrogen atom, a halogen atom or a C _1-4 haloalkyl;

   R ₃ and R ₄ are the same as or different from each other, and are each independently a hydrogen atom, C _1-6 alkyl, substituted or unsubstituted C _3-8 cycloalkyl, substituted or unsubstituted C _3-8 heterocycloalkyl, substituted or C _5-10 heteroaryl containing at least one heteroatom selected from the group consisting of unsubstituted C _5-10 aryl, substituted or unsubstituted N, O and S, or R ₃ and R ₄ are one or more carbon atoms are linked together to form a substituted or unsubstituted C _3-8 heterocycloalkyl or a substituted or unsubstituted C _5-10 heteroaryl, wherein C _3-8 cycloalkyl, C _3-8 hetero At least one H of cycloalkyl, C _5-10 aryl, or C _5-10 heteroaryl is each independently substituted with a halogen atom or C _1-6 alkyl.
4. The method of claim 1, wherein in Formula 1, when any one of R ₃ and R ₄ is a hydrogen atom, the other is a substituted or unsubstituted C _5-10 aryl, or

   R ₃ and R ₄ are C _3-8 heterocycloalkyl containing substituted or unsubstituted N atoms or C _5-10 heteroaryl containing substituted or unsubstituted N atoms formed by linking together one or more carbon atoms. Phosphorus, a pharmaceutical composition.
5. The method of claim 1, wherein in Formula 1, when any one of R ₃ and R ₄ is a hydrogen atom, the other is a substituted or unsubstituted phenyl, wherein at least one H of the substituted phenyl is independently substituted with a halogen atom That is, the pharmaceutical composition.
6. According to claim 1, in formula 1

   

   is formed by connecting R ₃ and R ₄ together with one or more carbon atoms

   

   or

   

   ego,

   here, above

   

   or

   

   Is unsubstituted, or at least one H is each independently a halogen atom, C _1-4 Haloalkyl or C _1-6 Alkyl is substituted, the pharmaceutical composition.
7. The method of claim 6, wherein the

   

   or

   

   Is unsubstituted, or one or more H is each independently methyl, F, Cl, Br, I or CF ₃ That will be substituted with, a pharmaceutical composition.
8. The pharmaceutical composition according to claim 1, wherein the compound represented by Formula 1 is any one selected from the group consisting of compounds described below:

   N-[2-(4-methyl-1-piperidinyl)-2-oxoethyl]-N-[3-(trifluoromethyl)phenyl]benzenesulfonamide;

   N~1~-phenyl-N~2~-(phenylsulfonyl)-N~2~-[3-trifluoromethyl)phenyl]glycinamide;

   N~1~-(4-fluorophenyl)-N~2~-(phenylsulfonyl)-N~2~-[3-(trifluoromethyl)phenyl]glycinamide;

   N-(3,4-dichlorophenyl)-N-[2-(4-methyl-1-piperidinyl)-2-oxoethyl]benzenesulfonamide;

   N-(3-chloro-4-fluorophenyl)-N-[2-oxo-2-(1-piperidinyl)ethyl] benzenesulfonamide; and

   N-(3,4-dichlorophenyl)-N-[2-(3,4-dihydro-2(1H)-isoquinolinyl)-2-oxoethyl]benzenesulfonamide.
9. The pharmaceutical composition according to claim 1, wherein the compound represented by Formula 1 is a compound described below.

   N-[2-(4-methyl-1-piperidinyl)-2-oxoethyl]-N-[3-(trifluoromethyl)phenyl]benzenesulfonamide.
10. The pharmaceutical composition according to claim 1, wherein the compound represented by Formula 1 or a pharmaceutically acceptable salt thereof inhibits mutual binding between MLK3 and Aurora kinase.
11. The method of claim 1, wherein the cancer is breast cancer, brain cancer, nerve cancer, colorectal cancer, lung cancer, small cell lung cancer, stomach cancer, liver cancer, blood cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, skin or intraocular melanoma, cervical cancer, ovarian cancer Cancer, rectal cancer, perianal cancer, colon cancer, breast cancer, fallopian tube carcinoma, endometrial carcinoma, cervical cancer, vaginal cancer, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, Cancer of the urethra, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, renal cell carcinoma, renal pelvic carcinoma, CNS tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma, and pituitary adenoma. Any one selected from the group consisting of, a pharmaceutical composition.
12. The pharmaceutical composition according to claim 1, wherein the cancer is breast cancer.
13. The pharmaceutical composition according to claim 12, wherein the breast cancer is triple negative breast cancer or HER2 positive breast cancer.
14. The pharmaceutical composition according to claim 12, wherein the breast cancer is triple negative breast cancer.
15. A method for preventing or treating cancer comprising administering a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof to a subject in need thereof:

    [Formula 1]

    

    In Formula 1,

    R ₁ and R ₂ are the same as or different from each other, and each independently represents a hydrogen atom, a halogen atom, C _1-4 haloalkyl or C _1-6 alkyl;

    R ₃ and R ₄ are the same as or different from each other, and are each independently a hydrogen atom, C _1-6 alkyl, substituted or unsubstituted C _3-8 cycloalkyl, substituted or unsubstituted C _3-8 heterocycloalkyl, substituted or C _5-10 heteroaryl containing at least one heteroatom selected from the group consisting of unsubstituted C _5-10 aryl, substituted or unsubstituted N, O and S, or R ₃ and R ₄ are one or more carbon atoms are linked together to form a substituted or unsubstituted C _3-8 heterocycloalkyl or a substituted or unsubstituted C _5-10 heteroaryl, wherein C _3-8 cycloalkyl, C _3-8 hetero At least one H of cycloalkyl, C _5-10 aryl, or C _5-10 heteroaryl may each independently be substituted with a halogen atom or C _1-6 alkyl;

    p and q are each independently an integer of 0 to 5;

    n is any one of the integers of 1-3.
16. The method of claim 15, wherein the cancer is breast cancer, brain cancer, nerve cancer, colorectal cancer, lung cancer, small cell lung cancer, stomach cancer, liver cancer, blood cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, skin or intraocular melanoma, cervical cancer, ovarian cancer Cancer, rectal cancer, perianal cancer, colon cancer, breast cancer, fallopian tube carcinoma, endometrial carcinoma, cervical cancer, vaginal cancer, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, Cancer of the urethra, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, renal cell carcinoma, renal pelvic carcinoma, CNS tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma, and pituitary adenoma. Any one selected from the group consisting of, method.
17. Use of the compound represented by formula (1) or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the prevention or treatment of cancer:

    [Formula 1]

    

    In Formula 1,

    R ₁ and R ₂ are the same as or different from each other, and each independently represents a hydrogen atom, a halogen atom, C _1-4 haloalkyl or C _1-6 alkyl;

    R ₃ and R ₄ are the same as or different from each other, and are each independently a hydrogen atom, C _1-6 alkyl, substituted or unsubstituted C _3-8 cycloalkyl, substituted or unsubstituted C _3-8 heterocycloalkyl, substituted or C _5-10 heteroaryl containing at least one heteroatom selected from the group consisting of unsubstituted C _5-10 aryl, substituted or unsubstituted N, O and S, or R ₃ and R ₄ are one or more carbon atoms are linked together to form a substituted or unsubstituted C _3-8 heterocycloalkyl or a substituted or unsubstituted C _5-10 heteroaryl, wherein C _3-8 cycloalkyl, C _3-8 hetero At least one H of cycloalkyl, C _5-10 aryl, or C _5-10 heteroaryl may each independently be substituted with a halogen atom or C _1-6 alkyl;

    p and q are each independently an integer of 0 to 5;

    n is any one of the integers of 1-3.
18. The method of claim 17, wherein the cancer is breast cancer, brain cancer, nerve cancer, colorectal cancer, lung cancer, small cell lung cancer, stomach cancer, liver cancer, blood cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, skin or intraocular melanoma, cervical cancer, ovarian cancer Cancer, rectal cancer, perianal cancer, colon cancer, breast cancer, fallopian tube carcinoma, endometrial carcinoma, cervical cancer, vaginal cancer, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, Cancer of the urethra, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, renal cell carcinoma, renal pelvic carcinoma, CNS tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma, and pituitary adenoma. Any one selected from the group consisting of, use.
19. A composition comprising a compound represented by Formula 1 or a pharmaceutically acceptable salt thereof for use in preventing or treating cancer:

    [Formula 1]

    

    In Formula 1,

    R ₁ and R ₂ are the same as or different from each other, and each independently represents a hydrogen atom, a halogen atom, C _1-4 haloalkyl or C _1-6 alkyl;

    R ₃ and R ₄ are the same as or different from each other, and are each independently a hydrogen atom, C _1-6 alkyl, substituted or unsubstituted C _3-8 cycloalkyl, substituted or unsubstituted C _3-8 heterocycloalkyl, substituted or C _5-10 heteroaryl containing at least one heteroatom selected from the group consisting of unsubstituted C _5-10 aryl, substituted or unsubstituted N, O and S, or R ₃ and R ₄ are one or more carbon atoms are linked together to form a substituted or unsubstituted C _3-8 heterocycloalkyl or a substituted or unsubstituted C _5-10 heteroaryl, wherein C _3-8 cycloalkyl, C _3-8 hetero At least one H of cycloalkyl, C _5-10 aryl, or C _5-10 heteroaryl may each independently be substituted with a halogen atom or C _1-6 alkyl;

    p and q are each independently an integer of 0 to 5;

    n is any one of the integers of 1-3.
20. The method of claim 19, wherein the cancer is breast cancer, brain cancer, nerve cancer, colorectal cancer, lung cancer, small cell lung cancer, stomach cancer, liver cancer, blood cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, skin or intraocular melanoma, cervical cancer, ovarian cancer Cancer, rectal cancer, perianal cancer, colon cancer, breast cancer, fallopian tube carcinoma, endometrial carcinoma, cervical cancer, vaginal cancer, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, Cancer of the urethra, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, renal cell carcinoma, renal pelvic carcinoma, CNS tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma, and pituitary adenoma. Which one is selected from the group consisting of, the composition.

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