Pyrazinocarbazole derivative or pharmaceutically acceptable salt thereof, and use thereof

Abstract

The present invention relates to a pyrazinocarbazole derivative or a pharmaceutically acceptable salt thereof, and a composition for preventing or treating cancer, comprising the derivative as an active ingredient. The pyrazinocarbazole derivative of the present invention can significantly inhibit the proliferation and growth of cancer cells by efficiently inhibiting YAP-TEAD binding in the Hippo pathway, which is known as a very important signaling system for the development and growth of cancer cells, and thus can be used for the prevention or treatment of cancer, preferably for the prevention and treatment of breast cancer.

Description

Pyrazinocarbazole derivatives or pharmaceutically acceptable salts thereof and their uses

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

[0002] Yes-associated protein (YAP) and its close homolog, WW-domain-containing transcription regulator 1 (WWTR1), are well known as oncogenes. As transcriptional co-activators, YAP and TAZ bind to DNA-binding transcription factors to activate the transcription and subsequent modification activities of target genes. Among the many transcription factors that bind to YAP and TAZ, the transcriptional enhancer factor (TEA)-domain (TEAD) family primarily mediates the major modification activities of YAP and TAZ, including activities related to cell proliferation, invasion, and metastasis [B. Zhao et al., TEAD mediates YAP-dependent gene induction and growth control, Genes Dev. 22 (2008) 1962-1971.]. Since YAP and TEAD cannot be directly administered as drugs, significant interest is focused on developing chemicals that interfere with the protein-protein interactions between them. Genetic and pharmacological intervention in the TEAD-YAP / TAZ interaction has been reported to significantly attenuate tumor formation in YAP-dependent cancer models [Y. Liu-Chittenden et al., Genetic and pharmacological disruption of the TEAD-YAP complex suppresses the oncogenic activity of YAP, Genes Dev. 26 (2012) 1300-1305.]. Additionally, verteporfin, an FDA-approved drug used as a photosensitizer in photodynamic therapy for macular degeneration, has been successfully repurposed as a YAP-TEAD inhibitor. However, verteporfin is a non-specific protein crosslinker, and its specificity for YAP-TEAD is uncertain.

[0003] Accordingly, the inventors designed and synthesized a novel YAP-TEAD inhibitor comprising a novel pyrazinocarbazole derivative, confirmed its activity against YAP / TAZ-TEAD through reporter analysis, and confirmed that it inhibits cancer cell growth, thereby completing and providing the present invention.

[0004] The technical problem that the present invention aims to solve is to provide a pyrazinocarbazole derivative or a pharmaceutically acceptable salt thereof.

[0005] The technical problem that the present invention aims to solve is to provide a method for preparing the pyrazinocarbazole derivative or a pharmaceutically acceptable salt thereof.

[0006] In addition, another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of cancer comprising the pyrazinocarbazole derivative or a pharmaceutically acceptable salt thereof as an active ingredient.

[0007] However, the technical problems that the present invention aims to solve are not limited to those mentioned above, and other unmentioned problems will be clearly understood by a person skilled in the art from the description below.

[0008] To solve the above problem, the present invention provides a pyrazinocarbazole derivative represented by the following [Chemical Formula 1] or a pharmaceutically acceptable salt thereof:

[0009] [Chemical Formula 1]

[0010]

[0011] In the above chemical formula 1,

[0012] R 1 It is either a sulfonyl or a hydrogen, and

[0013] The above sulfonyl is further substituted with any one selected from a C3-C6 cycloalkyl group, a C2-C6 heterocycloalkyl group, a C3-C6 aryl group, and a C3-C6 heteroaryl group, and

[0014] In the case where the above sulfonyl group is substituted with a C3-C6 heteroaryl group, the above heteroaryl group is additionally substituted with a methyl ester group, and

[0015] R 2 is hydrogen, C1-C6 alkyl, benzyloxy, substituted or unsubstituted phenoxy, and -NR 3 R 4 It is one of the selected ones, and

[0016] In the case where the above phenoxy group is substituted, it is substituted with a methoxy group, and

[0017] R 3 and R 4 Each is independently selected from the group consisting of hydrogen, adamantane-2-yl, (pyrrolidin-1-yl)pyridine-2-yl, and methyl butyrate, and

[0018] X is CH2 or NH.

[0019] According to one side, in the above [Chemical Formula 1],

[0020] R 1 is either a substituted sulfonyl and a hydrogen, and

[0021] In the case where the above sulfonyl is substituted, piperidine or It may have been replaced with one of them.

[0022] According to one side, in the above [Chemical Formula 1],

[0023] R 2 is hydrogen, methyl, benzyloxy, methoxyphenoxy, phenoxy, -NR 3 R 4 It is any one selected from the group consisting of, and

[0024] R 3 and R 4 Each independently hydrogen, adamanthan-2-yl, (pyrrolidin-1-yl)pyridin-2-yl and It can be any one selected from the group consisting of

[0025] According to one aspect, a pyrazinocarbazole derivative represented by [Chemical Formula 1] is characterized as being any one selected from the group consisting of the following compounds, or a pharmaceutically acceptable salt thereof:

[0026] [Chemical Formula 1-1]

[0027]

[0028] [Chemical Formula 1-2]

[0029]

[0030] [Chemical Formula 1-3]

[0031]

[0032] [Chemical Formula 1-4]

[0033]

[0034] [Chemical Formula 1-5]

[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] According to one aspect, the pyrazinocarbazole derivative may be characterized by inhibiting YAP-TEAD interaction.

[0045] According to one aspect, the pharmaceutically acceptable salt of the pyrazinocarbazole derivative may be characterized as being one or more selected from the group consisting of hydrochloride, bromate, sulfate, phosphate, nitrate, citrate, acetate, lactate, tartrate, maleate, gluconate, succinate, formate, trifluoroacetate, oxalate, fumarate, glutarate, adipose salt, methanesulfonate, benzenesulfonate, p-toluenesulfonate, camphorsulfonate, sodium salt, potassium salt, lithium salt, calcium salt, and magnesium salt, and preferably may be a hydrochloride.

[0046] According to another embodiment of the present invention, a pharmaceutical composition for the prevention or treatment of cancer is provided, comprising any one of the above-mentioned pyrazinocarbazole derivatives or a pharmaceutically acceptable salt thereof as an active ingredient.

[0047] According to one aspect, the cancer may be one or more selected from 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, thymic cancer, urethral cancer, and bronchial cancer, and preferably may be colorectal cancer.

[0048]

[0049] In addition, the present invention provides a method for preventing or treating cancer comprising the step of administering the pyrazinocarbazole derivative or a pharmaceutically acceptable salt thereof to an individual.

[0050] In addition, the present invention provides the use of the pyrazinocarbazole derivative or a pharmaceutically acceptable salt thereof for the manufacture of a drug for the prevention or treatment of cancer.

[0051] In addition, the present invention provides a method for diagnosing cancer comprising the step of administering the pyrazinocarbazole derivative or a pharmaceutically acceptable salt thereof to an individual.

[0052] In addition, the present invention provides the use of the pyrazinocarbazole derivative or a pharmaceutically acceptable salt thereof for the manufacture of a drug for cancer diagnosis.

[0053] The present invention relates to a pyrazinocarbazole derivative or a pharmaceutically acceptable salt thereof and a composition for the prevention or treatment of cancer comprising said derivative as an active ingredient. The pyrazinocarbazole derivative of the present invention exhibits an anticancer effect by inhibiting the YAP-TEAD interaction of the Hippo pathway, and thus can be used for the prevention or treatment of cancer.

[0054] The effects of the present invention are not limited to those mentioned above, and other unmentioned effects will be clearly understood by a person skilled in the art from the description below.

[0055] Figure 1 shows the cell death effect confirmed by administering the compound of the present invention to a colon cancer epithelial cell line.

[0056] Figure 2 shows IC based on the results of apoptosis in human colorectal cancer epithelial cell lines 50 It confirmed.

[0057] Figure 3 shows the measured activity of the YAP / TAZ-TEAD reporter gene.

[0058] Figure 4 shows the results of the anticancer efficacy evaluation experiment according to the concentration of the above composition in a colorectal cancer xenograft mouse model.

[0059] Figure 5 shows the toxicity evaluation of PD70 drugs at different concentrations in a colorectal cancer xenograft mouse model.

[0060] The inventors completed the present invention by conducting thorough studies on pyrazinocarbazole derivatives or pharmaceutically acceptable salts thereof and confirming the YAP-TEAD inhibitory activity of said derivatives.

[0061] More specifically, it was confirmed that the above-mentioned pyrazinocarbazole derivative or its pharmaceutically acceptable salt can inhibit the proliferation of cancer cell lines by inhibiting the binding of YAP-TEAD protein.

[0062] To solve the above problem, the present invention provides a pyrazinocarbazole derivative represented by the following [Chemical Formula 1] or a pharmaceutically acceptable salt thereof:

[0063] [Chemical Formula 1]

[0064]

[0065] In the above chemical formula 1,

[0066] R 1 It is either a sulfonyl or a hydrogen, and

[0067] The above sulfonyl is further substituted with any one selected from a C3-C6 cycloalkyl group, a C2-C6 heterocycloalkyl group, a C3-C6 aryl group, and a C3-C6 heteroaryl group, and

[0068] In the case where the above sulfonyl group is substituted with a C3-C6 heteroaryl group, the above heteroaryl group is additionally substituted with a methyl ester group, and

[0069] R 2 is hydrogen, C1-C6 alkyl, benzyloxy, substituted or unsubstituted phenoxy, and -NR 3 R 4 It is one of the selected ones, and

[0070] In the case where the above phenoxy group is substituted, it is substituted with a methoxy group, and

[0071] R 3 and R 4 Each is independently selected from the group consisting of hydrogen, adamantane-2-yl, (pyrrolidin-1-yl)pyridine-2-yl, and methyl butyrate, and

[0072] X is CH2 or NH.

[0073] In the present invention, the term “substitution” refers to a reaction that replaces an atom or atomic group contained in a molecule of a compound with another atom or atomic group.

[0074] In the present invention, the term “alkyl” refers to a saturated substituent consisting only of carbon and hydrogen without multiple bonds, and -C n H 2n+1 It means having the general formula of

[0075] In the present invention, the term “cycloalkyl” refers to a substituent in which the carbon skeleton of the alkyl group is arranged in a structure including a single ring, and -C n H 2n-1 It means having the general formula of

[0076] In the present invention, the term “heterocycloalkyl” means that at least one of the atoms constituting the ring of the cycloalkyl group has a heteroatom of N, O, or S.

[0077] In the present invention, the term "aryl" means an unsaturated aromatic ring compound having 3 to 12 carbon atoms having a single ring (e.g., phenyl) or a plurality of condensed rings (e.g., naphthyl). Examples of such aryl groups include, but are not limited to, phenyl, naphthyl.

[0078] In the present invention, the term “heteroaryl” refers to a single ring or a plurality of condensed rings having at least one of the atoms constituting the ring as a heteroatom of N, O, or S. Examples of such heteroaryl groups include, but are not limited to, pyridyl groups, pyrimidinyl groups, pyrazinyl groups, oxazolyl groups, furyl groups, thiophenyl groups, etc.

[0079] In the present invention, the term “sulfonyl” refers to a substituent having the general formula RS(=O)2-R′, where R may be the core of the pyrazinocarbazole derivative of the present invention, and R’ may be any one selected from a C3-C6 cycloalkyl group, a C2-C6 heterocycloalkyl group, a C3-C6 aryl group, and a C3-C6 heteroaryl group.

[0080] In the present invention, “benzyloxy” means -O-CH2-Ph. The above-mentioned Ph is a phenyl group.

[0081] In the present invention, “phenoxy” means -O-Ph. The phenoxy may be further substituted with a methoxy group.

[0082] In the present invention, “methyl butyrate” means -(CH2)3-(C=O)-O-CH3.

[0083] As a preferred embodiment of the present invention, the pyrazinocarbazole derivative represented by [Chemical Formula 1] may be any one selected from the group consisting of the following compounds:

[0084] [Chemical Formula 1-1]

[0085]

[0086] [Chemical Formula 1-2]

[0087]

[0088] [Chemical Formula 1-3]

[0089]

[0090] [Chemical Formula 1-4]

[0091]

[0092] [Chemical Formula 1-5]

[0093]

[0094] [Chemical Formula 1-8]

[0095]

[0096] [Chemical Formula 1-9]

[0097]

[0098] [Chemical Formula 1-10]

[0099]

[0100] [Chemical Formula 1-11]

[0101]

[0102] The pyrazinocarbazole derivative of the present invention or a pharmaceutically acceptable salt thereof effectively inhibits the YAP-TEAD binding in the Hippo pathway, which is known as a signaling system very important for the development and growth of cancer cells, thereby significantly inhibiting the proliferation and growth of cancer cells; thus, the pyrazinocarbazole derivative or a pharmaceutically acceptable salt thereof may be used as a pharmaceutical composition for the prevention or treatment of cancer comprising the pyrazinocarbazole derivative or the pharmaceutically acceptable salt thereof as an active ingredient.

[0103] In the present invention, the term “pharmaceuticalally acceptable salt” refers to a formulation of a compound that does not cause severe irritation to an organism to which the compound is administered and does not impair the biological activity and physical properties of the compound. The pharmaceutically acceptable salt may be obtained by reacting the compound of the present invention with an inorganic acid such as hydrochloric acid, bromic acid, sulfuric acid, nitric acid, phosphoric acid, sulfonic acid such as methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, or an organic carboxylic acid such as tartaric acid, formic acid, citric acid, acetic acid, trichloroacetic acid, trifluoroacetic acid, capric acid, isobutanoic acid, malonic acid, succinic acid, phthalic acid, gluconic acid, benzoic acid, lactic acid, fumaric acid, maleic acid, salicylic acid, etc. In addition, the compound of the present invention may be obtained by reacting it with a base to form salts such as ammonium salts, alkali metal salts such as sodium or potassium salts, alkaline earth metal salts such as calcium or magnesium salts, salts of organic bases such as dicyclohexylamine, N-methyl-D-glucarmine, tris(hydroxymethyl)methylamine, and amino acid salts such as arginine and lysine.

[0104] In addition, the above pyrazinocarbazole derivative or its pharmaceutically acceptable salt may include not only the pharmaceutically acceptable salt but also all salts, hydrates, and solvates that can be prepared by conventional methods.

[0105] In addition, the present invention may provide a method for the prevention, treatment, and / or diagnosis of cancer comprising the step of administering the pyrazinocarbazole derivative or a pharmaceutically acceptable salt thereof to an individual.

[0106] In the present invention, the term "prevention" refers to any act of suppressing or delaying the occurrence, spread, or recurrence of cancer by administering the composition of the present invention, and the term "treatment" refers to any act of improving or beneficially altering the symptoms of the said disease by administering the composition of the present invention.

[0107] In the present invention, the term "pharmaceutical composition" means one prepared for the purpose of preventing or treating a disease, and can be used by being formulated into various forms according to conventional methods. For example, it can be formulated into oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, and syrups, and can be used by being formulated into external preparations, suppositories, and sterile injectable solutions.

[0108] In the present invention, "included as an active ingredient" means that the corresponding ingredient is included in an amount necessary or sufficient to realize the desired biological effect. In actual application, the amount included as an active ingredient is determined as an amount for treating the target disease, taking into account factors that do not cause other toxicities, and may vary depending on various factors such as, for example, the disease or condition being treated, the form of the composition administered, the size of the subject, or the severity of the disease or condition. A person skilled in the art to which the present invention pertains can empirically determine the effective amount of an individual composition without involving excessive experimentation.

[0109] In addition, the pharmaceutical composition of the present invention may include one or more pharmaceutically acceptable carriers in addition to the active ingredients described above, depending on each formulation.

[0110] The above-mentioned pharmaceutically acceptable carrier may be saline solution, sterile water, Ringer's solution, buffered saline solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and mixtures of one or more of these components, and may further include other conventional additives such as antioxidants, buffers, and bacteriostatic agents as needed. Additionally, by additionally adding diluents, dispersants, surfactants, binders, and lubricants, it may be formulated into injectable formulations such as aqueous solutions, suspensions, and emulsions, or into pills, capsules, granules, or tablets. Furthermore, it may be preferably formulated according to each disease or component by appropriate methods in the art or by using methods disclosed in Remington's Pharmaceutical Science (Mack Publishing Company, Easton, PA).

[0111]

[0112] The composition of the present invention may be administered orally or parenterally in a pharmaceutically effective amount according to the intended method, and the term “pharmaceutically effective amount” of the present invention means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment and that does not cause side effects, and the effective dose level may be determined based on factors including the patient’s health status, severity, drug activity, sensitivity to the drug, method of administration, time of administration, route of administration and elimination rate, duration of treatment, drugs used in combination or concurrently, and other factors well known in the medical field.

[0113] Accordingly, the pharmaceutical composition of the present invention may be administered to an individual to prevent, treat, and / or diagnose cancer, and said cancer may be 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, thymic cancer, urethral cancer, or bronchial cancer, etc., but is not limited thereto, preferably may be a cancer in which YAP or TEAD is overexpressed, and a non-limiting example thereof is colorectal cancer.

[0114] In the present invention, the term “individual” is not limited to mammals such as livestock or humans that require prevention, treatment, and / or diagnosis of cancer, but preferably may be a human.

[0115] The pharmaceutical composition of the present invention can be formulated into various forms for administration to individuals, and representative formulations for parenteral administration are injectable formulations, preferably isotonic aqueous solutions or suspensions. Injectable formulations can be prepared according to techniques known in the art using suitable dispersants or wetting agents and suspending agents. For example, each component can be dissolved in saline solution or buffer solution to form an injectable formulation. Additionally, formulations for oral administration include, for example, ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, and wafers, and these formulations may contain, in addition to the active ingredient, a diluent (e.g., lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and / or glycine) and a lubricant (e.g., silica, talc, stearic acid and its magnesium or calcium salts and / or polyethylene glycol). The above tablet may include a binder such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidine, and optionally may further include a disintegrant such as starch, agar, alginic acid or its sodium salt, an absorbent, a coloring agent, a flavoring agent and / or a sweetener. The above formulation may be prepared by conventional mixing, granulation, or coating methods.

[0116] In addition, the pharmaceutical composition of the present invention may further include adjuvants such as preservatives, hydrating agents, emulsification promoters, salts or buffers for osmotic pressure regulation, and other therapeutically useful substances, and may be formulated according to conventional methods.

[0117] The pharmaceutical composition according to the present invention may be administered via various routes including oral, transdermal, subcutaneous, intravenous, or intramuscular, and the dosage of the active ingredient may be appropriately selected according to various factors such as the route of administration, the patient's age, gender, body weight, and severity of the patient. In addition, the composition of the present invention may be administered in combination with known compounds that can enhance the desired effect.

[0118] The pharmaceutical composition according to the present invention may be administered to humans and animals by route of administration, either orally or parenterally, such as intravenously, subcutaneously, intranasally, or intraperitoneally. Oral administration includes sublingual application. Parenteral administration includes injection methods such as subcutaneous injection, intramuscular injection, and intravenous injection, as well as drip methods.

[0119] In the pharmaceutical composition of the present invention, the total effective amount of the pyrazinocarbazole derivative or its pharmaceutically acceptable salt according to the present invention may be administered to a patient as a single dose, or administered via a fractionated treatment protocol in which multiple doses are administered over a long period. Although the content of the active ingredient in the pharmaceutical composition of the present invention may vary depending on the severity of the disease, it may typically be administered repeatedly several times a day at an effective dose of 100 μg to 3,000 mg per single dose for adults. However, the effective dose for the patient may be determined by considering various factors such as the patient's age, weight, health status, gender, severity of the disease, diet, and excretion rate, as well as the route of administration and frequency of treatment.

[0120] In addition, the pharmaceutical composition according to the present invention is not particularly limited in its formulation, route of administration, and method of administration as long as it exhibits the effects of the present invention, and the pharmaceutical composition of the present invention may additionally include a known anticancer agent as an active ingredient in addition to the pyrazinocarbazole derivative or a pharmaceutically acceptable salt thereof, and may be used in combination with other treatments known for the treatment of these diseases.

[0121]

[0122] The terms used in the embodiments are for illustrative purposes only and should not be interpreted as intended to be limiting. Singular expressions include plural expressions unless the context clearly indicates otherwise. In this specification, terms such as "comprising" or "having" are intended to indicate the existence of the features, numbers, steps, actions, components, parts, or combinations thereof described in the specification, and should be understood as not precluding the existence or addition of one or more other features, numbers, steps, actions, components, parts, or combinations thereof.

[0123] Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as generally understood by those skilled in the art to which the embodiments pertain. Terms such as those defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning in the context of the relevant technology, and should not be interpreted in an ideal or overly formal sense unless explicitly defined in this application.

[0124] The present invention is capable of various modifications and may have various embodiments. Specific embodiments are illustrated in the drawings and described in detail in the detailed description below. However, this is not intended to limit the present invention to specific embodiments, and it should be understood that it includes all modifications, equivalents, and substitutions that fall within the spirit and scope of the present invention. In describing the present invention, detailed descriptions of related prior art are omitted if it is determined that such detailed descriptions may obscure the essence of the present invention.

[0125]

[0126] Preparation Example 1. Synthesis of the compound of the present invention

[0127] The compounds of the present invention are as shown in Table 1 below. They were synthesized according to the following manufacturing examples.

[0128]

[0129] Preparation Example 1-1. General synthesis method

[0130]

[0131] Method 1: iPr2NEt (2.0 equivalents) was added at room temperature to a solution of pyrlindol 1 (1.0 equivalents) and sulfonyl chloride (1.1 equivalents) dissolved in CH3CN (0.1 M). After stirring for a specified time, the reaction mixture was diluted with ethyl acetate, acidified with 1N hydrochloric acid, and then extracted with ethyl acetate. The combined organic layer was dried with MgSO4 and concentrated under reduced pressure. The residue was purified by flash column chromatography (EtOAc / n-hexane) using silica gel to obtain PD38 and PD41 compounds.

[0132]

[0133] Method 2: Amine (1.5 equivalents), Pd(OAc)2 (0.1 equivalents), Xphos (0.1 equivalents), and NaOtBu (1.2 equivalents) were added at room temperature to a solution of Compound 2 (1.0 equivalents) dissolved in toluene (0.1 M). After stirring at 100 °C for a specified time, the reaction mixture was cooled and diluted with ethyl acetate, then acidified with a saturated NH4Cl solution and extracted with ethyl acetate. The combined organic layer was dried with NaSO4 and concentrated under reduced pressure. The residue was purified by flash column chromatography (EtOAc / n-hexane) using silica gel to obtain the intermediate and final compounds.

[0134]

[0135] Method 3: Methyl 4-bromobutyric acid ester (1.2 equivalents) and KOtBu (2.0 equivalents) were added at room temperature to a 0.1 M DMF solution of the intermediate (1.0 equivalent) prepared in Method 2. After stirring for a specified time under an N2 atmosphere, the reaction mixture was diluted with ethyl acetate, acidified with a saturated NH4Cl solution, and then extracted with ethyl acetate. The combined organic layer was dried with NaSO4 and concentrated under reduced pressure. The residue was purified by flash column chromatography (EtOAc / n-hexane) using silica gel to obtain the N-alkylation intermediate.

[0136]

[0137] Method 4: Pd / C (0.2 equivalents) and 4.0 M hydrochloric acid (1.0 equivalent) were added at room temperature to a solution of the N-alkylation intermediate (1.0 equivalent) dissolved in MeOH / CH2Cl2 (4:1, 0.1 M) or EtOH. After stirring for a specified time, the reaction mixture was diluted with CH2Cl2 and filtered with MeOH / CH2Cl2. The combined organic layer was dried with MgSO4 and concentrated under reduced pressure. The residue was purified by flash column chromatography (EtOAc / n-hexane) using silica gel to obtain the final compound.

[0138]

[0139] Preparation Example 1-2. Synthesis of 8-methyl-3-(piperidin-1-ylsulfonyl)-2,3,3a,4,5,6-hexahydro-1H-pyrazino[3,2,1-jk]carbazole (PD38)

[0140]

[0141] The reaction was carried out for 5 hours at a scale of 0.20 mmol according to Method 1 above. The residue was purified by flash column chromatography (EtOAc / n-hexane=1:1) to obtain brown solid PD38 (0.37 mmol, 82 mg, 67%).

[0142] 1H NMR (600 MHz, CDCl3) δ 7.29 (s, 1H), 7.15 (d,J= 8.2 Hz, 1H), 7.03 (dd,J= 8.3, 1.6 Hz, 1H), 4.74 - 4.62 (m, 1H), 4.19 (ddd,J= 11.9, 6.7, 3.2 Hz, 1H), 3.96 (ddd,J= 11.3, 6.9, 3.6 Hz, 1H), 3.69 (ddd,J= 13.5, 6.9, 3.2 Hz, 1H), 3.60 (ddd,J= 13.4, 6.7, 3.6 Hz, 1H), 3.24 (q,J= 5.7 Hz, 4H), 2.78 (dd,J= 15.7, 6.4 Hz, 1H), 2.71 - 2.63 (m, 1H), 2.64 - 2.54 (m, 1H), 2.46 (s, 3H), 2.24 - 2.06 (m, 1H), 1.98 - 1.83 (m, 1H), 1.77 - 1.61 (m, 6H), 1.57 (q,J= 3.3 Hz, 2H); 13 C NMR (150 MHz, CDCl3)δ136.04, 132.47, 129.07, 128.04, 123.11, 118.64, 109.57, 108.86, 54.84, 46.89, 46.12, 43.38, 30.27, 25.50, 23.87, 22.43, 21.57, 20.44; HRMS (EI+) calcd for C 20 H 27 N3O2S (M + ) 373.1824, found 373.1819.

[0143]

[0144] 제조예 1-3. Methyl-3-((8-methyl-1,2,3a,4,5,6-hexahydro-3H-pyrazino[3,2,1-jk]carbazol-3- yl)sulfonyl)thiophene-2-carboxylate (PD41)의 합성

[0145]

[0146] The reaction was carried out for 24 hours at a scale of 0.28 mmol according to Method 1 above. The residue was purified by flash column chromatography (EtOAc / n-hexane = 1:2) to obtain PD41 (0.26 mmol, 75 mg, 94%) in a white solid state.

[0147] 1 H NMR (600 MHz, CDCl3)δ8.19 (d,J= 1.1 Hz, 1H), 7.64 (t,J= 8.4 Hz, 1H), 7.42 - 7.37 (m, 2H), 7.32 - 7.26 (m, 2H), 7.24 - 7.18 (m, 1H), 7.11 (dd,J= 8.5, 1.0 Hz, 1H), 6.94 (dd,J= 8.5, 1.0 Hz, 1H), 5.79 (d,J= 4.4 Hz, 1H), 5.70 (dd,J= 4.4, 1.1 Hz, 1H), 3.80 (s, 3H); 13 C NMR (150 MHz, CDCl3)δ178.6, 160.2, 158.5, 152.0, 140.5, 134.3, 128.6, 128.0, 127.5, 126.8, 114.6, 110.3, 106.5, 70.8, 56.6; HRMS (EI+) calcd for C 21 H 22 N2O4S2(M + ) 430.1021, found 404.1044.

[0148]

[0149] Preparation Example 1-4. Synthesis of 9-methyl-2,3,3a,4,5,6-hexahydro-1H-2,4,6a-triazafluoranthene (PD68)

[0150]

[0151] The above compound PD68 was synthesized through the following process.

[0152]

[0153] Step 1: Synthesis of Benzyl 8-bromo-1,3,4,5-tetrahydro-2H-pyrido[4,3-b]indole-2-carboxylate(a-1)

[0154] Concentrated hydrochloric acid (0.1 mL) was slowly added at 0 °C to a solution of 4-bromophenylhydrazine (1.92 g, 8.57 mmol) and 1-Cbz-4-piperidone (2.00 g, 8.57 mmol) dissolved in ethanol (50 mL). After stirring at 90 °C for 8 hours, the reaction mixture was terminated with water and extracted with ethyl acetate. The organic layer was dried with NaSO4 and concentrated under reduced pressure. The mixture was filtered with methanol using Whatman filter paper, yielding a-1 (3.10 g, 94%) as a pale yellow solid as the residue.

[0155] 1 H NMR (600 MHz, CDCl3) δ9.36 (s, 1H), 7.79 (m, 1H), 7.43 (dd,J= 8.8, 1.9 Hz, 1H), 7.34 (dd,J= 8.8, 0.6 Hz, 1H), 2.97 (t,J= 6.1 Hz, 2H), 2.68 (dd,J= 7.3, 5.7 Hz, 2H), 2.27 (p,J= 6.3 Hz, 2H). 13 C NMR (150 MHz, CDCl3) δ191.4, 136.3, 132.2, 130.0, 128.5, 127.7, 124.1, 114.1, 113.7, 38.3, 25.0, 21.4.

[0156]

[0157] Step 2: Synthesis of Benzyl 4-(benzylamino)-8-bromo-1,3,4,5-tetrahydro-2H-pyrido[4,3-b]indole-2-carbo xylate(a-2)

[0158] Under dark conditions, t-butyl alcohol (18.5 mL, 8.32 mmol) and glacial acetic acid (12.2 mL, 22.71 mmol) were slowly added to a commercial household bleach solution (250 mL) at 0 °C. After stirring for 3 minutes at room temperature, the reaction mixture was first washed with 10% Na2CO3 and then washed with water. The organic layer was dried with calcium chloride (500 mg) and filtered. The filtrate yielded a-2 (2.21 g, 82%) as a pale yellow liquid.

[0159] 1 H NMR (600 MHz, DMSO-d6) δ11.22 (s, 1H), 7.68 (d,J= 2.0 Hz, 1H), 7.33 (m, 10H), 7.20 (m, 2H), 5.14 (d,J= 11.9 Hz, 2H), 4.61 (m, 2H), 3.89 (m, 5H). 13 C NMR (150 MHz, DMSO-d6) δ155.02 (d,J= 41.9 Hz), 140.69, 136.96, 136.62 (d,J= 38.1 Hz), 134.75, 128.41, 128.11, 127.93, 127.83, 127.61, 126.62, 123.56, 120.28, 113.33, 111.15, 66.40, 50.27 (d,J= 20.1 Hz), 49.90 (d,J= 13.6 Hz), 45.61, 40.98 (d,J= 17.9 Hz).

[0160]

[0161] Step 3: Synthesis of Benzyl 4-benzyl-9-bromo-3a,4,5,6-tetrahydro-1H-2,4,6a-triazafluoranthene-2(3H)-carboxylate(a-3)

[0162] NaH (405 mg, 16.89 mmol) and ethylene ditosylate (2.09 g, 22.71 mmol) were added at 0 °C to a solution of a-2 (2.0 g, 5.63 mmol) dissolved in DMSO (20 mL). After stirring at room temperature for 5 hours, the reaction mixture was terminated with a saturated NH4Cl solution and extracted with ethyl acetate. The organic layer was dried with NaSO4 and concentrated under reduced pressure. The mixture was filtered through methanol and a small amount of acetone using Whatman filter paper to obtain a-3 (1.91 g, 89%) as a white solid.

[0163] 1 H NMR (600 MHz, CDCl3) δ7.58 (m, 1H), 7.38 (m, 8H), 7.11 (d,J= 8.6 Hz, 1H), 5.24 (m, 2H), 5.09 (dd,J= 54.3, 15.5 Hz, 1H), 4.87 (ddd,J= 124.3, 12.2, 4.7 Hz, 1H), 4.29 (dd,J= 70.7, 14.4 Hz, 1H), 4.19 (dd,J= 118.5, 14.6 Hz, 1H), 4.05 (dd,J= 11.0, 3.3 Hz, 2H), 3.73 (m, 1H), 3.66 (m, 1H), 3.41 (dd,J= 29.0, 13.4 Hz, 1H), 3.18 (m, 1H), 2.88 (dd,J= 12.2, 10.2 Hz, 1H), 2.71 (m, 1H). 13 C NMR (150 MHz, CDCl3) δ156.4, 137.7, 136.6, 136.3, 135.2, 129.1, 128.7, 128.7, 128.3, 128.2, 128.2, 127.6, 124.3, 121.2, 121.0, 113.3, 111.0, 104.2, 67.8, 58.3, 56.8, 49.3, 45.6, 42.9, 41.6.

[0164]

[0165] Step 4: Synthesis of 9-methyl-2,3,3a,4,5,6-hexahydro-1H-2,4,6a-triazafluoranthene (PD68)

[0166]

[0167] Trimethylboroxine (19 mg, 0.16 mmol), K2CO3 (32 mg, 0.24 mmol), and Pd(dppf)Cl2 (5.7 mg, 0.008 mmol) were added to a solution of a-3 (30 mg, 0.08 mmol) dissolved in 1,4-dioxane (1.5 mL). The mixture was then stirred in a microwave at 130 °C for 1 hour. The reaction mixture was diluted with ethyl acetate, the organic layer was washed with water and saturated salt water, and then dried with MgSO4. The solvent was removed under reduced pressure, and the solution was purified by flash column chromatography using silica gel (ethyl acetate / hexane / methanol = 1:2:0.1) to obtain a white solid a-4 (92%). Subsequently, a reaction to remove the protecting group of a-4 was carried out for 0.5 hours according to Method 4 above at a scale of 0.03 mmol. The residue was purified by flash column chromatography (EtOAc / n-hexane / methanol = 1:2:0.1) to obtain pale yellow solid PD68 (9 mg, 96%).

[0168] 1 H NMR (600 MHz, MeOD) δ7.25 (d,J= 9.1 Hz, 2H), 7.13 - 6.88 (m, 1H), 4.37 - 4.29 (m, 1H), 4.28 - 4.22 (m, 2H), 4.20 (dd,J= 11.4, 4.5 Hz, 1H), 3.70 (td,J= 11.8, 5.0 Hz, 1H), 3.64 (dd,J= 11.5, 5.2 Hz, 1H), 3.49 (dd,J= 13.1, 5.0 Hz, 1H), 3.37 (dd,J= 12.5, 4.5 Hz, 1H), 2.87 (t,J=11.1 Hz, 1H), 2.41 (s, 3H).

[0169]

[0170] Preparation Example 1-5. Synthesis of N-(((1R,3S,5r,7r)-adamantan-2-yl)methyl)-2,3,3a,4,5,6-hexahydro-1H-pyrazino[3,2,1-jk]carbazol-8-amine(PD70)

[0171]

[0172] The above PD70 compound was synthesized through the following process.

[0173]

[0174] Step 1: Synthesis of N-(((1R,3S,5r,7r)-Adamantan-2-yl)methyl)-3-benzyl-2,3,3a,4,5,6-hexahydro-1H-pyrazino[3,2,1-jk]carbazol-8-amine(b-4)

[0175] The reaction was carried out for 16 hours at a scale of 0.26 mmol according to Method 2 above. The residue was purified by flash column chromatography (EtOAc / n-hexane = 1:4) to obtain b-4 (0.21 mmol, 122 mg, 80%) in a white solid state.

[0176] 1H NMR (600 MHz, CDCl3) δ7.37 (d,J= 7.2 Hz, 2H), 7.32 (t,J= 7.5 Hz, 2H), 7.27 (m, 1H), 7.01 (d,J= 8.5 Hz, 1H), 6.68 (d,J= 2.2 Hz, 1H), 6.57 (dd,J= 8.5, 2.3 Hz, 1H), 4.28 (d,J= 13.2 Hz, 1H), 3.97 (m, 1H), 3.63 (m, 1H), 3.49 (m, 1H), 3.20 (d,J= 13.3 Hz, 1H), 3.10 (m, 1H), 2.84 (s, 2H), 2.75 (dd,J= 15.5, 6.2 Hz, 1H), 2.63 (m, 3H), 2.38 (m, 1H), 2.18 (m, 1H), 1.86 (m, 1H), 1.70 (m, 7H), 1.60 (m, 8H). 13 C NMR (150 MHz, CDCl3) δ143.6, 139.0, 135.7, 132.0, 129.2, 128.5, 127.2, 110.5, 109.8, 107.5, 101.0, 59.7, 58.4, 57.5, 50.2, 43.0, 41.0, 37.4, 34.0, 29.9, 28.6, 28.5, 22.5, 20.8.

[0177]

[0178] Step 2: Synthesis of N-(((1R,3S,5r,7r)-adamantan-2-yl)methyl)-2,3,3a,4,5,6-hexahydro-1H-pyrazino[3,2,1-jk]carbazol-8-amine(PD70)

[0179] The reaction was carried out for 0.5 hours at a scale of 0.05 mmol according to general procedure 4. The residue was purified by flash column chromatography (EtOAc / n-hexane = 1:2) to obtain 15a as a white solid (0.05 mmol, 18 mg, 90%).

[0180] 1H NMR (600 MHz, CDCl3) δ7.06 (d,J= 8.5 Hz, 1H), 6.68 (d,J= 2.2 Hz, 1H), 6.58 (dd,J= 8.6, 2.3 Hz, 1H), 4.05 (m, 1H), 3.93 (m, 1H), 3.60 (m, 1H), 3.45 (m, 1H), 3.36 (m, 1H), 2.84 (s, 2H), 2.74 (dd,J= 15.4, 6.3 Hz, 1H), 2.63 (m, 1H), 2.15 (m, 3H), 1.87 (m, 2H), 1.71 (m, 7H), 1.62 (d,J= 2.9 Hz, 7H), 1.44 (m, 1H). 13 C NMR (150 MHz, CDCl3) δ143.8, 136.4, 132.5, 129.0, 110.5, 109.9, 107.1, 100.9, 58.3, 52.7, 44.1, 43.8, 41.0, 37.3, 34.0, 30.5, 29.8, 28.6, 22.6, 21.1.

[0181]

[0182] Preparation Example 1-6. Synthesis of N-((6-(Pyrrolidin-1-yl)pyridin-2-yl)methyl)-2,3,3a,4,5,6-hexahydro-1H-pyrazino[3,2,1-jk]carbazol-8-amine (PD71)

[0183]

[0184] The above compound PD71 was synthesized through the following process.

[0185]

[0186] Step 1: Synthesis of 3-Benzyl-N-((6-(pyrrolidin-1-yl)pyridin-2-yl)methyl)-2,3,3a,4,5,6-hexahydro-1H-pyrazino[3,2,1-jk]carbazol-8-amine(b-5)

[0187] The reaction was carried out for 16 hours at a scale of 0.26 mmol according to Method 2 above. The residue was purified by flash column chromatography (EtOAc / n-hexane = 1:4) to obtain b-5 (0.20 mmol, 93 mg, 75%) in a pale yellow solid state.

[0188] 1 H NMR (600 MHz, CDCl3) δ7.38 (m, 3H), 7.34 (m, 2H), 7.28 (m, 1H), 7.06 (d,J= 8.5 Hz, 1H), 6.79 (d,J= 2.2 Hz, 1H), 6.68 (dd,J= 8.5, 2.2 Hz, 1H), 6.57 (d,J= 7.1 Hz, 1H), 6.22 (dd,J= 8.3 Hz, 1H), 4.33 (s, 2H), 4.28 (d,J= 13.3 Hz, 1H), 3.98 (m, 1H), 3.66 (m, 1H), 3.50 (m, 5H), 3.20 (d,J= 13.3 Hz, 1H), 3.11 (m, 1H), 2.76 (m, 1H), 2.62 (m, 1H), 2.39 (m, 1H), 2.02 (m, 4H), 1.88 (m, 1H), 1.60 (m, 1H), 1.29 (m, 1H). 13 C NMR (150 MHz, CDCl3) δ157.1, 157.1, 142.8, 139.0, 137.4, 135.6, 132.1, 129.2, 129.1, 129.1, 128.5, 128.4, 127.1, 110.9, 109.8, 108.9, 107.5, 104.5, 101.5, 59.7, 57.5, 50.9, 50.2, 46.7, 42.9, 28.5, 25.7, 22.5, 20.8.

[0189]

[0190] Step 2: Synthesis of N-((6-(Pyrrolidin-1-yl)pyridin-2-yl)methyl)-2,3,3a,4,5,6-hexahydro-1H-pyrazino[3,2,1-jk]carbazol-8-amine (PD71)

[0191] The reaction was carried out according to Method 4 above. The residue was purified by flash column chromatography (EtOAc / n-hexane = 1:2 to 1:1) to obtain pale yellow solid PD71 (0.04 mmol, 16 mg, 85%).

[0192] 1 H NMR (600 MHz, CDCl3) δ7.36 (m, 2H), 7.32 (m, 2H), 7.02 (dd,J= 8.4, 0.6 Hz, 1H), 6.79 (d,J= 1.9 Hz, 1H), 6.60 (dd,J= 8.4, 2.2 Hz, 1H), 4.28 (d,J= 13.3 Hz, 1H), 3.98 (m, 1H), 3.65 (m, 1H), 3.49 (m, 2H), 3.20 (d,J= 13.3 Hz, 1H), 3.11 (m, 1H), 2.73 (dd,J= 15.6, 6.3 Hz, 1H), 2.63 (m, 2H), 2.39 (m, 1H), 2.19 (m, 2H), 1.87 (m, 2jH), 0.88 (m, 2H). 13 C NMR (150 MHz, CDCl3) δ129.2, 128.5, 127.2, 111.5, 109.8, 104.3, 59.7, 57.5, 53.6, 50.2, 43.0, 29.9, 28.5, 22.4, 20.7.

[0193]

[0194] Preparation Example 1-7. Synthesis of Methyl 4-((((1R,3S,5r,7r)-adamantan-2-yl)methyl)(2,3,3a,4,5,6-hexahydro-1H-pyrazino[3,2,1-jk]carbazol-8-yl)amino)butanoate (PD73)

[0195]

[0196] The above compound PD73 was synthesized through the following process.

[0197]

[0198] Step 1: Synthesis of Methyl 4-((((1R,3S,5r,7r)-adamantan-2-yl)methyl)(3-benzyl-2,3,3a,4,5,6-hexahydro-1H-pyrazino[3,2,1-jk]carbazol-8-yl)amino)butanoate(b-6)

[0199] The reaction was carried out for 24 hours at a scale of 0.13 mmol according to Method 3 above. The residue was purified by flash column chromatography (EtOAc / n-hexane = 1:4) to obtain b-6 (0.08 mmol, 43 mg, 59%) in a white solid state.

[0200] 1 H NMR (600 MHz, CDCl3) δ 7.37 - 7.31 (m, 5H), 7.06 (d,J= 8.8 Hz, 1H), 6.94 (s, 1H), 6.84 (d,J= 8.7 Hz, 1H), 4.28 (d,J= 13.3 Hz, 1H), 3.98 (d,J= 9.8 Hz, 1H), 3.66 (s, 3H), 3.70 - 3.61 (m, 1H), 3.52 - 3.46 (m, 1H), 3.30 (t,J= 7.4 Hz, 2H), 3.19 (d,J= 13.3 Hz, 1H), 3.10 (d,J=11.5 Hz, 2H), 2.95 (s, 2H), 2.86 - 2.58 (m, 4H), 2.38 (d,J= 11.7 Hz, 1H), 2.33 (t,J= 7.4 Hz, 2H), 2.20 (m, 1H), 1.92 (s, 3H), 1.83 (t,J= 7.3) Hz, 2H), 1.67 (d,J= 12.2 Hz, 3H), 1.61 (m, 5H), 1.55 (s, 6H). 13C NMR (150 MHz, CDCl3) δ 174.2, 144.8, 139.1, 135.8, 132.0, 129.1, 129.1, 128.8, 128.5, 127.2, 113.3, 109.4, 107.5, 106.0, 66.1, 59.7, 57.4, 54.7, 51.6, 50.2, 43.0, 41.7, 41.0, 37.3, 37.3, 36.9, 31.7, 28.7, 28.6, 28.5, 22.5, 22.1, 20.8.

[0201]

[0202] Step 2: Synthesis of Methyl 4-((((1R,3S,5r,7r)-adamantan-2-yl)methyl)(2,3,3a,4,5,6-hexahydro-1H-pyrazino[3,2,1-jk]carbazol-8-yl)amino)butanoate(PD73)

[0203] The reaction was carried out for 0.5 hours at a scale of 0.05 mmol according to Method 4 above. The residue was purified by flash column chromatography (EtOAc / n-hexane = 1:3) to obtain PD73 (0.05 mmol, 21 mg, 90%) in a white solid state.

[0204] 1 H NMR (600 MHz, CDCl3) δ 7.10 (d,J= 8.6 Hz, 1H), 6.96 - 6.77 (m, 2H), 4.15 (ddd,J= 29.8, 19.3, 8.3 Hz, 3H), 3.93 - 3.71 (m, 3H), 3.71 - 3.64 (m, 3H), 3.49 (s, 1H), 3.33 (s, 2H), 2.97 (s, 2H), 2.77 (dd,J= 15.8, 5.8 Hz, 1H), 2.66 (t,J= 15.4 Hz, 1H), 2.43 - 2.28 (m, 3H), 2.24 - 2.17 (m, 1H), 2.03 (d,J= 11.8 Hz, 1H), 1.93 (s, 4H), 1.83 (q,J= 7.3 Hz, 2H), 1.71 - 1.59 (m, 8H).

[0205]

[0206] Preparation Example 1-8. Synthesis of N-(((1R,3S,5r,7r)-Adamantan-2-yl)methyl)-2,3,3a,4,5,6-hexahydro-1H-2,4,6a-triazafluoranthen-9-amine (PD76)

[0207]

[0208] The above compound PD76 was synthesized through the following process.

[0209]

[0210] Step 1: Synthesis of Benzyl 9-((((1R,3S,5r,7r)-adamantan-2-yl)methyl)amino)-4-benzyl-3a,4,5,6-tetrahy dro-1H-2,4,6a-triazafluoranthene-2(3H)-carboxylate(c-4)

[0211] The reaction was carried out for 16 hours at a scale of 0.19 mmol according to Method 2 above. The residue was purified by flash column chromatography (EtOAc / n-hexane = 1:4) to obtain c-4 (0.15 mmol, 90 mg, 79%) in a white solid state.

[0212] 1H NMR (600 MHz, CDCl3) δ 7.46 - 7.37 (m, 5H), 7.37 - 7.26 (m, 5H), 7.09 (d,J= 9.2 Hz, 1H), 6.87 (t,J= 7.7 Hz, 2H), 5.24 (d,J= 9.7 Hz, 3H), 5.09 (dd,J= 39.9, 15.1 Hz, 1H), 4.97 - 4.71 (m, 1H), 4.42 - 4.30 (m, 1H), 4.26 - 4.19 (m, 1H), 4.00 (dd,J= 10.8, 3.8 Hz, 1H), 3.73 - 3.58 (m, 7H), 3.46 - 3.27 (m, 3H), 3.12 (d,J= 11.3 Hz, 1H), 3.01 - 2.85 (m, 3H), 2.74 - 2.69 (m, 1H), 2.46 - 2.39 (m, 1H), 2.35 - 2.29 (m, 2H), 2.03 - 1.98 (m, 1H), 1.93 (s, 3H), 1.86 - 1.81 (m, 2H), 1.67 (d,J= 13.0 Hz, 4H), 1.62 - 1.59 (m, 5H), 1.58 - 1.53 (m, 7H). 13 C NMR (150 MHz, CDCl3) δ 174.1, 156.5, 145.0, 136.9, 129.0, 128.7, 128.6, 128.3, 128.1, 127.4, 113.2, 109.8, 104.7, 104.1, 67.6, 67.0, 65.5, 63.0, 58.3, 56.8, 54.3, 51.9, 51.6, 49.6, 43.0, 41.6, 41.0, 37.3, 37.2, 37.1, 31.7, 28.6, 28.6, 24.2, 21.8.

[0213]

[0214] 단계 2:N-(((1R,3S,5r,7r)-Adamantan-2-yl)methyl)-2,3,3a,4,5,6-hexahydro-1H-2,4,6a-triazafluoranthen-9-amine(PD76)의 합성

[0215] The reaction was carried out for 0.5 hours at a scale of 0.05 mmol according to Method 4 above. The residue was purified by flash column chromatography (EtOAc / n-hexane / methanol = 1:2:0.1) to obtain pale yellow solid PD76 (0.04 mmol, 17 mg, 88%).

[0216] 1 H NMR (600 MHz, MeOD-d4) δ 7.76 - 7.63 (m, 2H), 7.43 - 7.30 (m, 1H), 4.66 - 4.51 (m, 3H), 4.03 (tt,J= 11.5, 5.3 Hz, 2H), 3.95 - 3.87 (m, 2H), 3.79 (dd,J= 13.0, 5.0 Hz, 1H), 3.38 (t,J= 11.1 Hz, 1H), 3.12 (s, 2H), 2.66 (s, 3H), 2.08 (s, 3H), 1.84 (d,J= 12.7 Hz, 3H), 1.80 - 1.71 (m, 9H). 13 C NMR (150 MHz, MeOD) δ 132.8, 127.0, 121.5, 119.9, 118.0, 113.2, 113.2, 66.7, 62.9, 57.8, 50.0, 49.8, 44.3, 43.8, 42.3, 42.2, 40.7, 37.5, 34.0, 29.5, 29.5.

[0217]

[0218] Experimental Example 1. Confirmation of anticancer effect in colorectal cancer epithelial cell lines

[0219] Cell viability was analyzed after treating two human colorectal cancer epithelial cell lines, HT29 and HCT116, which have opposing genetic mutations, with the compound PD70 at various concentrations, and the results are shown in Figure 1. Both cell lines were cultured using McCoy's 5A (Modified) medium supplemented with 10% Fetal Bovine Serum (FBS) and 1% Antibiotic-Antimycotic (A / A). Cultured cells were 1 x 10⁶ 4The cells were seeded into a 96-well plate and cultured for 24 hours. Subsequently, the pyrindol derivative PD70 was administered at various concentrations, with a total of 100 µl per well. During drug treatment, a medium free of FBS was used to eliminate interference from the assay reagent. After 24 hours of drug treatment, the plates were left at room temperature for 20 minutes to ensure a constant temperature in each well, and CellTiter-Glo® Luminescent reagent was added and reacted for 20 minutes. Afterward, luminescence was measured using a GloMax® Microplate Luminometer. The results showed that cell viability decreased in both cell types at lower concentrations compared to the control pyrindol construct, PD00. In other words, the PD70 compound of the present invention demonstrated excellent apoptotic efficacy even at low concentrations.

[0220] In addition, IC based on cell viability at different concentrations 50 The process of deriving is shown in FIG. 2. The compound PD70 of the present invention is IC in HT28 50 The value is 9.104nM, IC in HCT116 50 The value was 5.070 nM, showing a very low IC50 value.

[0221]

[0222] Experimental Example 2. Analysis of the inhibitory effect on YAP / TAZ-TEAD complex formation

[0223] Two human colorectal cancer epithelial cell lines with opposing genetic mutations, HT29 and HCT116, were cultured in McCoy's 5A (Modified) medium supplemented with 10% Fetal Bovine Serum (FBS) and 1% Antibiotic-Antimycotic (A / A). The cultured cells were 5 x 10⁶ 5After seeding the cells into a 12-well plate, they were cultured for 24 hours. Subsequently, overexpression of Gal4-TEAD4, 5 x upstream activation sequence (UAS)-luciferase reporter, and Renilla was induced using OPTI-MEM medium and Lipofectamine reagent. After 6 hours, the pyrindol derivative drug PD70 was added at a dose of 1 ml per well and treated for 24 hours. Luciferase activity was measured using the Dual-Glo luciferase assay system (E2940). As shown in Figure 3, the composition PD70 showed significantly lower binding affinity than the control pyrindol construct PD00 in both cell types, confirming a high inhibitory effect on the complex.

[0224]

[0225] Experimental Example 3. Confirmation of anticancer effect using a colorectal cancer xenograft mouse model

[0226] 5 x 10 human colorectal cancer epithelial cell lines HT29 per cell 6 Colorectal cancer was induced by subcutaneous injection into the hind leg flank / dorsal region of 7-week-old male Balb / c nude strain mice, mixed with Matrigel at a 1:1 ratio. From 7 days later, when colorectal cancer formation began to be observed (size: minimum 40–50 mm 3 (Time reached) The above composition PD70 drug was administered intraperitoneally a total of 8 times at 2-day intervals, divided into concentrations of 1 mg, 2.5 mg, 5 mg, and 10 mg, and tumor size was measured. When the size of the colon cancer was obtained by euthanasia after the last oral administration and compared, the most significant reduction in colon cancer size was confirmed in the group administered PD70 drug at a concentration of 10 mg (Fig. 4).

[0227]

[0228] Experimental Example 4. Evaluation of Toxicity in a Colorectal Cancer Xenograft Mouse Model

[0229] When comparing the changes in body weight and the weight changes of five major organs (Heart, Lung, Spleen, Kidney, Liver) of colorectal cancer-induced mice treated with the above composition PD70 drug at different concentrations (1 mg, 2.5 mg, 5 mg, 10 mg), no toxicity was observed (Fig. 5).

[0230]

[0231] Although the embodiments have been described above with reference to the limited drawings, those skilled in the art can apply various technical modifications and variations based on the above. For example, suitable results may be achieved even if the described techniques are performed in a different order than described, and / or if the components of the described system, structure, device, circuit, etc. are combined or assembled in a form different from described, or replaced or substituted by other components or equivalents.

[0232] Therefore, other embodiments, other manufacturing examples, and equivalents to the claims also fall within the scope of the claims set forth below.

Claims

1. A pyrazinocarbazole derivative represented by the following [Chemical Formula 1] or a pharmaceutically acceptable salt thereof: [Chemical Formula 1] In the above chemical formula 1, R 1 It is either a sulfonyl or a hydrogen, and The above sulfonyl is further substituted with any one selected from a C3-C6 cycloalkyl group, a C2-C6 heterocycloalkyl group, a C3-C6 aryl group, and a C3-C6 heteroaryl group, and In the case where the above sulfonyl group is substituted with a C3-C6 heteroaryl group, the above heteroaryl group is additionally substituted with a methyl ester group, and R 2 is hydrogen, C1-C6 alkyl, benzyloxy, substituted or unsubstituted phenoxy, and -NR 3 R 4 It is one of the selected ones, and In the case where the above phenoxy group is substituted, it is substituted with a methoxy group, and R 3 and R 4 Each is independently selected from the group consisting of hydrogen, adamantane-2-yl, (pyrrolidin-1-yl)pyridine-2-yl, and methyl butyrate, and X is CH2 or NH.

2. In Paragraph 1, In the above [Chemical Formula 1], R 1 is either a substituted sulfonyl and a hydrogen, and In the case where the above sulfonyl is substituted, piperidine or A pyrazinocarbazole derivative substituted with any one of the above, or a pharmaceutically acceptable salt thereof.

3. In Paragraph 1, In the above [Chemical Formula 1], R 2 is hydrogen, methyl, benzyloxy, methoxyphenoxy, phenoxy, -NR 3 R 4 It is any one selected from the group consisting of, and R 3 and R 4 Each independently hydrogen, adamanthan-2-yl, (pyrrolidin-1-yl)pyridin-2-yl and A pyrazinocarbazole derivative or a pharmaceutically acceptable salt thereof, selected from the group consisting of 4. The pyrazinocarbazole derivative or a pharmaceutically acceptable salt thereof, wherein the pyrazinocarbazole derivative represented by [Chemical Formula 1] is selected from the group consisting of the following compounds: [Chemical Formula 1-1] [Chemical Formula 1-2] [Chemical Formula 1-3] [Chemical Formula 1-4] [Chemical Formula 1-5] [Chemical Formula 1-8] [Chemical Formula 1-9] [Chemical Formula 1-10] [Chemical Formula 1-11] 5. In Paragraph 1, The above pyrazinocarbazole derivative is a pyrazinocarbazole derivative or a pharmaceutically acceptable salt thereof characterized by inhibiting YAP-TEAD interaction.

6. In Paragraph 1, A pyrazinocarbazole derivative or a pharmaceutically acceptable salt thereof, characterized in that the pharmaceutically acceptable salt of the above-mentioned pyrazinocarbazole derivative is one or more selected from the group consisting of hydrochloride, bromate, sulfate, phosphate, nitrate, citrate, acetate, lactate, tartrate, maleate, gluconate, succinate, formate, trifluoroacetate, oxalate, fumarate, glutarate, adipose salt, methanesulfonate, benzenesulfonate, p-toluenesulfonate, camphorsulfonate, sodium salt, potassium salt, lithium salt, calcium salt, and magnesium salt.

7. In Paragraph 1, A pyrazinocarbazole derivative or a pharmaceutically acceptable salt thereof, characterized in that the pharmaceutically acceptable salt of the above pyrazinocarbazole derivative is a hydrochloride salt.

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

9. In Paragraph 8, A pharmaceutical composition for the prevention or treatment of cancer, characterized in that the above cancer is one or more selected from 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, thymic cancer, urethral cancer, and bronchial cancer.

10. In Paragraph 9, A pharmaceutical composition for the prevention or treatment of cancer, characterized in that the cancer is colorectal cancer.

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