Novel compound for treating cancer having KRAS g12d mutation, and composition for prevention or treatment of cancer using same

Abstract

The present invention relates to a novel compound that specifically binds to the KRAS G12D protein, and a pharmaceutical composition comprising same for the prevention or treatment of cancer having a KRAS G12D mutation.

Description

Novel compound for treating cancer with KRAS G12D mutation and composition for the prevention or treatment of cancer using the same

[0001] The present invention relates to a novel compound that specifically binds to the KRAS G12D protein and a pharmaceutical composition for the prevention or treatment of cancer containing the same. Specifically, the novel compound of the present invention that specifically binds to the KRAS G12D protein has excellent binding affinity to the KRAS G12D protein and has an inhibitory effect on cell growth of cancer containing the KRAS G12D mutation, thereby providing a pharmaceutical composition for the prevention or treatment of cancer using the same.

[0002] The MAPK / ERK signaling pathway regulates various cellular responses, including cell proliferation, differentiation, and apoptosis, by transmitting extracellular stimuli to the nucleus. The KRAS protein acts as an initiator of the MAPK / ERK signaling pathway and functions as a switch responsible for inducing cell division. In the inactive state of the KRAS (V-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog) protein, it binds to guanosine diphosphate (GDP) and effectively transmits negative signals to inhibit cell division. In response to extracellular signals, the KRAS protein undergoes allosteric activation, enabling the nucleotide exchange of GDP for guanosine triphosphate (GTP). In the GTP-bound active state of the KRAS protein, it recruits and activates other cellular signaling receptors as well as proteins necessary for the propagation of growth factor-induced signaling. Examples of proteins mobilized by KRAS-GTP are c-Raf and PI3-kinase. As a GTPase, KRAS converts bound GTP back to GDP, thereby returning itself to an inactive state and propagating signals to inhibit cell division. KRAS gain-of-function mutations exhibit an increased degree of GTP binding and a reduced ability to convert GTP to GDP. The result is increased MAPK / ERK signaling that promotes cancer cell growth. In other words, when amino acid substitutions occur due to mutations in the KRAS gene, KRAS remains in a constant state of activation due to impaired GTPase function or reduced responsiveness to GTPase-activating proteins, continuously sending signals downstream. This excessive signaling leads to carcinogenesis or the accelerated proliferation of cancer cells.

[0003] The KRAS gene is one of several genes associated with the development of non-small cell lung cancer, colorectal cancer, and pancreatic cancer, and KRAS mutations are the most commonly found oncogenic factors. Generally, it is known that approximately 25% of all cancers, about 30% of lung adenocarcinomas and colorectal cancers, and 80% of pancreatic cancers carry KRAS mutations. The mutations classified as the most common targets are KRAS G12C, KRAS G12V, and KRAS G12D.

[0004] Pancreatic cancer, primarily characterized by pancreatic ductal adenocarcinoma, is a cancer with a very poor prognosis, with a 5-year survival rate of less than 10% (CA Cancer J. Clin., 2016, 66, p.7-30), and approximately 460,000 new cases are reported annually worldwide (CA Cancer J. Clin., 2018, 68, p.394-424). While surgical intervention is the most effective treatment for pancreatic cancer, early detection is difficult, so the disease often metastasizes, making it difficult to expect therapeutic effects from surgery in many cases. When surgery is not performed, chemotherapy or radiation therapy is used, but the survival rate remains poor. Currently, FOLFRINOX therapy (a multidrug combination therapy adding levofolinate to three types of chemotherapy agents—5-FU, irinotecan, and oxaliplatin—is used as the standard therapy for pancreatic cancer; however, due to its strong toxicity, careful patient selection is required, such as limiting the prescribed patients to those with an ECOG Performance Status of 1 or lower (J. Clin. Oncol., 2018, 36, p.2545-2556). As for molecular targeted therapies, the epidermal growth factor receptor (EGFR) inhibitor erlotinib has been approved for combination therapy with gemcitabine; however, the extension of total survival is only about two weeks compared to gemcitabine alone, and satisfactory therapeutic effects have not been achieved, so there is still a need for a highly effective treatment (J. Clin. Oncol., 2007, 25, p.1960-1966).

[0005] Pancreatic ductal adenocarcinoma, the most common type of pancreatic cancer, is believed to develop from pancreatic intraepithelial neoplasia (PanIN) through stages ranging from a mild to a severe abnormal phase, and KRAS gene mutations are already observed in early-stage PanIN. Subsequently, abnormalities in tumor suppressor genes such as INK4A, p53, or SMAD4 occur, leading to malignancy (Nature Rev. Cancer, 2010, 10, p.683-695). Furthermore, mutations in the KRAS gene are observed in more than 90% of pancreatic ductal adenocarcinomas, with point mutations at codon 12 in KRAS exon 2 accounting for the majority (Cancer Cell 2017, 32, p.185-203). From this, KRAS plays an important role in the carcinogenesis and development of pancreatic cancer.

[0006] Against this background, the inventors have completed the present invention by confirming a novel compound that specifically binds to the KRAS G12D protein and an inhibitory effect on cell growth of cancers containing KRAS G12D mutations that exhibits excellent binding affinity to the KRAS G12D protein. In particular, it was confirmed that the inhibitory effect on the growth of pancreatic cancer cells is excellent and can resolve the problems of disease recurrence and resistance to therapeutic agents.

[0007] [Prior Art Literature]

[0008] [Patent Literature]

[0009] Korean Patent Publication No. 10-2023-0016158

[0010] The present invention relates to a novel compound that specifically binds to the KRAS G12D protein, and a pharmaceutical composition for the prevention or treatment of cancer comprising the same as an active ingredient, which has excellent binding affinity to the KRAS G12D protein and excellent cell growth inhibitory activity in cancers where the KRAS G12D mutation is present.

[0011] To solve the above problem, the present invention provides a compound of the following formula I, a solvate, a stereoisomer, or a pharmaceutically acceptable salt thereof:

[0012] [Chemical Formula I]

[0013]

[0014] In the above formula,

[0015] R1 and R2 are each independently H or -CF3, and

[0016] R3 to R7 are each independently H, F, Cl, -CF3, -NH2, -OH, -C 1-6 alkyl or -OC 1-6 It is an alkyl.

[0017] In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of cancer, comprising the above compound, solvate, stereoisomer, or pharmaceutically acceptable salt thereof as an active ingredient.

[0018] The compound, solvate, stereoisomer, or pharmaceutically acceptable salt thereof of Formula I according to the present invention is a novel compound that specifically binds to the KRAS G12D protein and has excellent binding affinity to the KRAS G12D protein. The compound, solvate, stereoisomer, or pharmaceutically acceptable salt thereof of Formula I has an inhibitory effect on cell growth in cancers containing the KRAS G12D mutation and can be utilized for the prevention or treatment of various cancers. Furthermore, a pharmaceutical composition comprising the compound, solvate, stereoisomer, or pharmaceutically acceptable salt thereof of Formula I can be usefully used for the prevention or treatment of cancer, and in particular for the prevention or treatment of pancreatic cancer.

[0019] Hereinafter, various embodiments of the present invention are described with reference to the accompanying drawings. The present invention is not limited to specific embodiments and should be understood to include various modifications, equivalents, and / or alternatives of the embodiments of the present invention. In connection with the description of the drawings, similar reference numerals may be used for similar components.

[0020] In this document, expressions such as "have," "can have," "include," or "can include" refer to the existence of the relevant feature (e.g., numerical values, functions, actions, or components, etc.) and do not exclude the existence of additional features.

[0021] In this document, expressions such as “A or B,” “at least one of A or / and B,” or “one or more of A or / and B” may include all possible combinations of items listed together. For example, “A or B,” “at least one of A and B,” or “at least one of A or B” may refer to cases including (1) at least one A, (2) at least one B, or (3) both at least one A and at least one B.

[0022] As used in this document, the expression "configured to" may be replaced, depending on the context, with, for example, "suitable for," "having the capacity to," "designed to," "adapted to," "made to," or "capable of." The term "configured to" does not necessarily mean "specifically designed to."

[0023] The terms used in this document are used merely to describe specific embodiments and are not intended to limit the scope of other embodiments. Singular expressions may include plural expressions unless the context clearly indicates otherwise. Terms used herein, including technical or scientific terms, may have the same meaning as generally understood by those skilled in the art described in this document. Terms used in this document that are defined in general dictionaries may be interpreted as having the same or similar meaning as they have in the context of the relevant technology, and are not to be interpreted in an ideal or overly formal sense unless explicitly defined in this document. In some cases, even terms defined in this document may not be interpreted to exclude the embodiments of this document.

[0024] The embodiments disclosed in this document are presented for the purpose of explaining and understanding the disclosed technical content and are not intended to limit the scope of the invention. Accordingly, the scope of this document should be interpreted to include all modifications or various other embodiments based on the technical concept of the invention.

[0025] Hereinafter, preferred embodiments of the present invention will be described in detail. Prior to this, terms and words used in this specification and claims should not be interpreted as being limited to their ordinary or dictionary meanings, and should be interpreted in a meaning and concept consistent with the technical spirit of the present invention, based on the principle that the inventor can appropriately define the concept of the terms to best describe his invention.

[0026] Therefore, it should be understood that the configurations of the embodiments described in this specification are merely some of the most preferred embodiments of the present invention and do not represent all of the technical ideas of the present invention, and that various equivalents and modifications that can replace them may exist at the time of filing this application.

[0027] Throughout the specification, when a part is described as "including" a certain component, this means that, unless specifically stated otherwise, it does not exclude other components but may include additional components.

[0028] The present invention will be described in detail below.

[0029]

[0030] The present invention relates to a compound represented by the chemical formula I, and more specifically, to a novel compound that specifically binds to the KRAS G12D protein and a pharmaceutical composition for the prevention or treatment of cancer containing the same as an active ingredient.

[0031] [Chemical Formula I]

[0032]

[0033] Unless otherwise noted, the terms used in the description and claims of the present invention have the meanings disclosed below.

[0034] In accordance with conventions used in the industry, in the chemical formula of this invention " It is used to indicate that a residue or substituent "R" is attached to the skeletal structure.

[0035] "Alkyl" is a hydrocarbon having primary, secondary, tertiary, and / or quaternary carbon atoms and includes a saturated aliphatic group that may be straight-chain, branched, cyclic, or a combination thereof. For example, an alkyl group has 1 to 20 carbon atoms (i.e., C1-C 20 alkyl), 1 to 10 carbon atoms (i.e., C1-C 10It may have alkyl), or 1 to 6 carbon atoms (i.e., C1-C6 alkyl). Unless otherwise defined, in a preferred embodiment, alkyl refers to C1-C6 alkyl. Examples of suitable alkyl groups include methyl (Me, -CH3), ethyl (Et, -CH2CH3), 1-propyl (n-Pr, n-propyl, -CH2CH2CH3), 2-propyl (i-Pr, i-propyl, -CH(CH3)2), 1-butyl (n-Bu, n-butyl, -CH2CH2CH2CH3), 2-methyl-1-propyl (i-Bu, i-butyl, -CH2CH(CH3)2), 2-butyl (s-Bu, s-butyl, -CH(CH3)CH2CH3), 2-methyl-2-propyl (t-Bu, t-butyl, -C(CH3)3), 1-pentyl (n-pentyl, -CH2CH2CH2CH2CH3), 2-pentyl (-CH(CH3)CH2CH2CH3), 3-pentyl (-CH(CH2CH3)2), and 2-methyl-2-butyl (-C(CH3)2CH2CH3), 3-methyl-2-butyl (-CH(CH3)CH(CH3)2), 3-methyl-1-butyl (-CH2CH2CH(CH3)2), 2-methyl-1-butyl (-CH2CH(CH3)CH2CH3), 1-hexyl (-CH2CH2CH2CH2CH2CH3), 2-hexyl (-CH(CH3)CH2CH2CH2CH3), 3-hexyl (-CH(CH2CH3)(CH2CH2CH3)), 2-methyl-2-pentyl (-C(CH3)2CH2CH2CH3), 3-methyl-2-pentyl (-CH(CH3)CH(CH3)CH2CH3), 4-methyl-2-pentyl (-CH(CH3)CH2CH(CH3)2), 3-methyl-3-pentyl (-C(CH3)(CH2CH3)2), Examples include, but are not limited to, 2-methyl-3-pentyl (-CH(CH2CH3)CH(CH3)2), 2,3-dimethyl-2-butyl (-C(CH3)2CH(CH3)2), 3,3-dimethyl-2-butyl (-CH(CH3)C(CH3)3), and octyl (-(CH2)7CH3).

[0036] Furthermore, the term “alkyl” used throughout the specification, examples, and claims is intended to include both unsubstituted and substituted alkyl groups, the latter of which refers to alkyl residues having substituents that replace hydrogen on one or more carbons of a hydrocarbon backbone, such as haloalkyl groups like trifluoromethyl and 2,2,2-trifluoroethyl.

[0037] Term "C x-y " or "C x -C y When used with chemical residues such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, it is considered to contain a group having x to y carbons in the chain. A CO alkyl group represents hydrogen when the group is at a terminal position and a bond when it is internal. For example, a (C1-C6)alkyl group contains 1 to 6 carbon atoms in the chain.

[0038] "Alkoxy" refers to a group having the formula -O-alkyl, in which an alkyl group as defined above is attached to a parent compound via an oxygen atom. The alkyl residue of the alkoxy group is, for example, 1 to 20 carbon atoms (i.e., C1-C 20 alkoxy), 1 to 12 carbon atoms (i.e., C1-C 12 alkoxy), 1 to 10 carbon atoms (i.e., C1-C 10 It may have an alkoxy group, or one to six carbon atoms (i.e., a C1-C6 alkoxy group). Examples of suitable alkoxy groups include, but are not limited to, methoxy (-O-CH3 or -OMe), ethoxy (-OCH2CH3 or -OEt), and t-butoxy (-OC(CH3)3 or -O-tBu).

[0039] "Alkenyl" has primary, secondary, tertiary, and / or quaternary carbon atoms, comprises linear, branched, and cyclic groups, or combinations thereof, and has one or more unsaturated regions, i.e., carbon-carbon sp² 2It is a hydrocarbon having a double bond. For example, an alkenyl group consists of 2 to 20 carbon atoms (i.e., C2-C 20 alkenyl), 2 to 12 carbon atoms (i.e., C2-C 12 alkenyl), 2 to 10 carbon atoms (i.e., C2-C 10 It may have alkenyl, or 2 to 6 carbon atoms (i.e., C2-C6 alkenyl). Examples of suitable alkenyl groups include, but are not limited to, vinyl (-CH=CH2), allyl (-CH2CH=CH2), cyclopentenyl (-C5H7), and 5-hexenyl (-CH2CH2CH2CH2CH=CH2).

[0040] "Alkynyl" is a hydrocarbon having primary, secondary, tertiary, and / or quaternary carbon atoms, comprising linear, branched, and cyclic groups, or combinations thereof, and having one or more carbon-carbon sp³ triple bonds. For example, an alkynyl group has 2 to 20 carbon atoms (i.e., C2-C 20 alkynyl), 2 to 12 carbon atoms (i.e., C2-C 12 alkynyl), 2 to 10 carbon atoms (i.e., C2-C 10 It may have alkynyl, or 2 to 6 carbon atoms (i.e., C2-C6 alkynyl). Examples of suitable alkynyl groups include acetylenyl (-C≡CH) and propynyl (-CH2C≡CH), but are not limited thereto.

[0041] As used herein, the term "aryl" comprises a substituted or unsubstituted monovalent or divalent aromatic hydrocarbon group that is monocyclic, bicyclic, or polycyclic, in which each atom of the ring is carbon. Preferably, the aryl ring is a 6- to 20-membered ring, a 6- to 14-membered ring, a 6- to 10-membered ring, or more preferably a 6-membered ring. The aryl group may be a polycyclic ring system having two or more cyclic rings in which two or more carbons are common to two adjacent rings, wherein one or more of the rings are aromatic, and, for example, the other cyclic rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocycloalkyl. Examples of aryl groups include benzene, naphthalene, phenanthrene, anthracene, indene, indan, phenol, aniline, etc.

[0042] The terms “carbocyclylalkyl”, “cycloalkylalkyl”, or “(cycloalkyl)alkyl” as used herein refer to an alkyl group substituted with a carbocycline group or a cycloalkyl group.

[0043] As used herein, the terms “carbocycle,” “carbocyclil,” “carbocyclic,” or “cycloalkyl” refer to a non-aromatic saturated or unsaturated, monovalent or divalent ring that may be monocyclic, bicyclic, or polycyclic, in which each atom of the ring is carbon. A cycloalkyl group may have 3 to 7 carbon atoms as a monocycle, 7 to 12 carbon atoms as a bicycle, and about 20 or fewer carbon atoms as a polycycle. A monocyclic cycloalkyl has 3 to 7 ring atoms, more typically 5 or 6 ring atoms. A bicyclic cycloalkyl may have 7 to 12 ring atoms and may be a fused ring system, a spirocyclic ring system, or a bridged ring system. In an exemplary cycloalkyl group, the atoms may be arranged in a bicyclo[4,5], [5,5], [5,6], or [6,6] system. In certain embodiments, the cycloalkyl contains 3 to 20 atoms, or 3 to 10 atoms, or more preferably 3 to 7 atoms. Examples of cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, etc. Unless otherwise specified, the cycloalkyl may be substituted by one or more of the substituents described herein.

[0044] The terms "heterocyclylalkyl" and "heterocycloalkyl" as used herein refer to alkyl groups substituted with heterocycloalkyl groups.

[0045] The terms “heterocyclil,” “heterocycle,” “heterocyclic,” and “heterocycloalkyl” refer to a substituted or unsubstituted, monovalent or divalent, saturated or partially saturated non-aromatic ring structure, preferably a 3- to 10-membered ring, more preferably a 3- to 7-membered ring, in which the ring structure comprises one or more heteroatoms, preferably 1 to 4 heteroatoms, more preferably 1 to 2 heteroatoms. The terms “heterocyclile,” “heterocycle,” “heterocyclic,” and “heterocycloalkyl” also comprise a polycyclic ring system having two or more cyclic rings in which two or more carbons are common to two adjacent rings, wherein one or more of the rings are heterocyclic, and for example, the other cyclic ring may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclic. Bicyclic and polycyclic heterocyclic ring systems may be fused, bridged, or spirocyclic ring systems. Substituted heterocycles comprise a heterocyclic ring substituted with any substituent disclosed herein, including a carbonyl group, for example. The heterocyclic group includes, for example, piperidine, piperazine, pyrrolidine, morpholine, lactone, lactam, etc.Additionally, exemplary heterocyclos include dihydropyridyl, dihydroindolyl, tetrahydropyridyl (piperidyl), tetrahydrothiophenyl, sulfur-oxidized tetrahydrothiophenyl, indolenyl, piperidinyl, 4-piperidinyl, pyrrolidinyl, 2-pyrrolidonyl, pyrrolinyl, tetrahydrofuranil, tetrahydroquinolinil, tetrahydroisoquinolinil, decahydroquinolinil, octahydroisoquinolinil, 6H-1,2,5-thiadiazinyl, 2H,6H-1,5,2-dithiazinyl, pyranil, cropnyl, xanthenyl, phenoxatinyl, 2H-pyrrolil, 3H-indolyl, 4H-quinolizinyl, phthalazinyl, naftiridinyl, quinoxalinyl, Examples include, but are not limited to, quinazolinil, cinnolinil, pteridinil, 4aH-carbazolyl, carbazolyl, β-carbolinil, phenanthridinil, acridinil, phenanthrolinil, phenazinil, phenothiazinil, furazanil, phenoxazinil, isochromanil, chromanil, imidazolidinil, imidazolidinil, pyrazolidinil, pyrazolidinil, piperazinil, methylpiperazinil, quinuclidinil, morpholinil, azabicyclo(2.1.1)hexanil, azacycloheptanil, 1-oxa-3-aza-cycloheptanil, azetidinil, aziridinil and oxazolidinil (each of which may be substituted or unsubstituted).

[0046] "Heteroaryl" refers to a substituted or unsubstituted monovalent or divalent aromatic group that is monocyclic, bicyclic, or polycyclic and contains one or more heteroatoms within the ring. Non-limiting examples of suitable heteroatoms that may be contained in the aromatic ring include oxygen, sulfur, and nitrogen. In a polycyclic heteroaryl ring system, the ring system has two or more cyclic rings in which two or more carbons are common to two adjacent rings, wherein one or more of the rings are heteroaromatic, and, for example, the other cyclic ring may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. Heterogroups include, for example, benzofuran, benzothiophen, pyrrole, furan, thiophene, imidazole, indole, isoindole, isoxazole, isothiaazole, oxazole, thiazole, quinoline, isoquinoline, pyrazol, pyridine, pyrazine, pyridazine, and pyrimidine, etc. (each of which may be substituted or unsubstituted).

[0047] The terms "halo" and "halogen" as used herein mean halogens and include chloro, fluoro, bromo, and iodine.

[0048] The present invention relates to compounds of the following formula I, solvates, stereoisomers, or pharmaceutically acceptable salts thereof.

[0049] [Chemical Formula I]

[0050]

[0051] In the above formula,

[0052] R1 and R2 are each independently H or -CF3, and

[0053] R3 to R7 are each independently H, F, Cl, -CF3, -NH2, -OH, -C 1-6 alkyl or -OC 1-6 It is an alkyl.

[0054] Specifically, the present invention comprises compounds of Formula I, solvates, stereoisomers, or pharmaceutically acceptable salts thereof, selected from the group consisting of compounds represented by the formulas in [Table 1] below, but not limited thereto.

[0055] [Table 1]

[0056]

[0057]

[0058]

[0059]

[0060]

[0061] In addition, specifically, the present invention relates to a pharmaceutical composition for the prevention or treatment of cancer, comprising as an active ingredient a compound of Formula I, a solvate, a stereoisomer, or a pharmaceutically acceptable salt thereof.

[0062] The term "cancer" is a collective term for diseases caused by cells that possess aggressive characteristics—dividing and proliferating while disregarding normal growth limits—invasive characteristics—infiltrating surrounding tissues—and metastatic characteristics—spreading to other parts of the body.

[0063] The above cancer may be a cancer containing tumor tissue in which a KRAS G12D mutation exists.

[0064] Mutations at codons 12 and 13 of the aforementioned KRAS gene lead to functional changes in the gene's product, the p21-ras protein. Consequently, this promotes cell growth and division by transmitting growth signals to the cell nucleus in excess of what is necessary, thereby participating in the carcinogenesis process. KRAS mutations can manifest in various ways depending on changes in location and sequence; representative examples include G12D, G12V, G12R, and G12C, where the glycine amino acid residue at the 12th position is substituted with aspartic acid, valine, arginine, or cysteine, or G13D, G13V, and G13H, where the glycine amino acid residue at the 13th position is substituted with aspartic acid, valine, or histidine. It has been reported that the oncological profile varies depending on the type of KRAS mutation, which can be used to assess the effectiveness of anticancer treatments such as cetuximab or panitummab, and consequently, lead to differences in patient survival rates. KRAS mutations are very common in human cancers, appearing in about 90% of pancreatic cancers, about 50% of colorectal cancers, and about 30% of non-small cell lung cancers, and most of these mutations are known to be concentrated in codons 12 and 13.

[0065] The above cancer may be one or more selected from the group consisting of breast cancer, lung cancer, stomach cancer, prostate cancer, uterine cancer, ovarian cancer, kidney cancer, pancreatic cancer, liver cancer, colorectal cancer, colon cancer, skin cancer, head and neck cancer, melanoma, and thyroid cancer, but is not limited thereto. Preferably, the above cancer may be pancreatic cancer, but is not limited thereto.

[0066] In addition, from the experimental results of the examples, the compound of Formula I of the present invention has the effect of inhibiting the growth of cancer cells, including tumor tissues containing KRAS G12D mutations, and can exhibit excellent therapeutic effects on various cancer-related diseases.

[0067] In this specification, the terms “treating” or “treating” mean suppressing a disease, e.g., suppressing a disease, condition, or disorder in an individual experiencing or exhibiting a pathology or sign of a disease, condition, or disorder, i.e., preventing further occurrence of the pathology and / or sign; or improving a disease, e.g., improving a disease, condition, or disorder in an individual experiencing or exhibiting a pathology or sign of a disease, condition, or disorder, i.e., reversing the pathology and / or sign, e.g., reducing the severity of the disease.

[0068] In this specification, the terms “preventing” or “prevention” mean preventing a disease, for example, preventing a disease, condition, or disorder in an individual who may have a predisposition to a disease, condition, or disorder but has not yet experienced or exhibited the pathology or signs of the disease.

[0069] In the present invention, the pharmaceutical composition may include conventionally pharmaceutically acceptable carriers, excipients, or additives. The pharmaceutical composition may be formulated according to conventional methods and may be prepared in various forms of oral administration such as tablets, pills, powders, capsules, syrups, emulsions, microemulsions, or forms of parenteral administration such as intramuscular, intravenous, or subcutaneous administration.

[0070] When the above pharmaceutical composition is prepared in the form of an oral formulation, examples of additives or carriers used include cellulose, calcium silicate, corn starch, lactose, sucrose, dextrose, calcium phosphate, stearic acid, magnesium stearate, calcium stearate, gelatin, talc, surfactants, suspending agents, emulsifiers, diluents, etc. When the pharmaceutical composition of the present invention is prepared in the form of an injectable formulation, examples of additives or carriers include water, saline solution, glucose aqueous solution, similar sugar aqueous solution, alcohol, glycol, ether (e.g., polyethylene glycol 400), oil, fatty acid, fatty acid ester, glyceride, surfactant, suspending agent, emulsifier, etc.

[0071] The dosage of the above pharmaceutical composition is an amount effective for the treatment or prevention of an individual or patient, and may be administered orally or parenterally as intended. When administered orally, it may be administered in an amount of 0.001 to 10 mg per kg of body weight per day based on the active ingredient, more specifically 0.1 to 10 mg; when administered parenterally, it may be administered in an amount of 0.01 to 10 mg per kg of body weight per day based on the active ingredient, more specifically 0.1 to 10 mg. It should be understood that the dosage for a specific individual or patient should be determined in light of various relevant factors such as the patient's body weight, age, gender, health status, diet, time of administration, method of administration, and severity of the disease, and that it may be appropriately adjusted by a professional. The above dosage is not intended to limit the scope of the present invention in any way. A physician or veterinarian with ordinary skills in the relevant technical field can easily determine and prescribe the required effective amount of the pharmaceutical composition. For example, a physician or veterinarian may start the dosage of the compound of the present invention used in a pharmaceutical composition at a level lower than that required to achieve the desired therapeutic effect, and gradually increase the dosage until the desired effect is achieved.

[0072] Additionally, the above pharmaceutical composition may be administered for tumor therapy in combination with chemotherapy, radiation therapy, immunotherapy, hormone therapy, bone marrow transplantation, stem cell replacement therapy, other biological therapies, surgical intervention, or combinations thereof. For example, it may be used as an adjuvant therapy in conjunction with other long-term treatment strategies, or to promote tumor regression in severe patients or to maintain the patient's condition after chemoprophylaxis.

[0073] The present invention will be described in more detail below through examples. These examples are intended solely to explain the present invention more specifically, and it will be obvious to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention.

[0074]

[0075] [Preparation Example]

[0076] Compounds 1 to 26 of the present invention were prepared according to the following preparation examples and embodiments.

[0077] Preparation Example 1: Preparation of Ethyl 4-Hydroxypyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-Carboxylate

[0078]

[0079] Preparation Example 1-1: Preparation of Methyl 3-aminofuro[2,3-b]pyridine-2-carboxylate

[0080] Cesium carbonate (260 g, 797 mmol, 1.84 eq) and 2-chloropyridine-3-carbonitrile (60.0 g, 433 mmol, 1.00 eq) were added to a solution of methyl 2-hydroxyacetate (46.6 g, 518 mmol, 1.20 eq) dissolved in dimethylformamide (800 mL) at 20°C, and the mixture was reacted at 80°C for 2 hours. The reaction mixture was stopped by adding 300 mL of water at 25°C, and extracted with ethyl acetate (100 mL, 3 times). The combined organic layer was washed with brine solution (100 mL, 2 times), dried with sodium sulfate, filtered, and concentrated under reduced pressure. The product was stirred with methanol (100 mL) at 20°C for 1 hour. methyl 3-aminofuro[2,3-b]pyridine-2-carboxylate (38.0 g, 173 mmol, 40.0% yield) was obtained as a yellow solid by concentrating under reduced pressure.

[0081] LC / MS: Rt = 0.257 min, m / z (M+H)+ = 193.0

[0082] Preparation Examples 1-2: Preparation of Ethyl 4-Hydroxypyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-Carboxylate

[0083] Ethyl cyanoformate (20.6 g, 207 mmol, 2.00 eq) and hydrochloric acid (15.0 mL) were added to a solution of methyl 3-aminofuro[2,3-b]pyridine-2-carboxylate (20.0 g, 104 mmol, 1.00 eq) in acetic acid (150 mL). The reaction mixture was stirred at 70°C for 12 hours. The pH of the reaction mixture was adjusted to 5 and filtered to obtain a crystalline solid. This was recrystallized and concentrated under vacuum to obtain ethyl 4-hydroxypyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxylate (17.0 g, 52.4 mmol, 50.4% yield) as a white solid.

[0084] LC / MS: Rt = 1.232 min, m / z (M+H)+ = 259.9

[0085] Preparation Example 2: Preparation of 4-Benzylpyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxylic acid

[0086]

[0087] Preparation Example 2-1: Preparation of Ethyl 4-Chloropyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxylate

[0088] Phosphoryl chloride (32.9 g, 214 mmol, 11.1 eq) was added to a mixture of ethyl 4-hydroxypyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxylate (5.00 g, 19.2 mmol, 1.00 eq) and DIEA (1.48 g, 11.4 mmol, 0.59 eq) at 0 °C and stirred at 70 °C for 5 hours. The reaction mixture was slowly added dropwise to 150 mL of water at 25 °C, then the solution was filtered, washed with 50 mL of water, and filtered to obtain a solid cake, which was dried under vacuum. The filtered filtrate was extracted with 450 mL of ethyl acetate (150 mL, 3 times). The combined organic layer was washed with 200 mL of brine solution, dried with sodium sulfate, filtered, and concentrated under reduced pressure, then combined with the first obtained solid cake to obtain ethyl 4-chloropyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxylate (3.20 g, 8.99 mmol, yield 46.6%) as a yellow solid.

[0089] Preparation Example 2-2: Preparation of 4-Benzylpyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxylic acid

[0090] A mixture of ethyl 4-chloropyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxylate (2.30 g, 8.28 mmol, 1.00 eq), 2-benzyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (2.71 g, 12.4 mmol, 1.50 eq), sodium carbonate (2.64 g, 24.9 mmol, 3.01 eq) and Pd(dppf)Cl2 (303 mg, 414 μmol, 0.05 eq) is added to dioxane (20.0 mL) and water (10.0 mL), degassed at 20°C, purged with nitrogen three times, and then reacted at 80°C for 13 hours. The reaction mixture was diluted with 100 mL of ethyl acetate and extracted with water (200 mL). The combined aqueous layer was adjusted to pH 4 with 1 M hydrochloric acid and extracted with 30.0 mL of ethyl acetate (10 mL, 3 times), the combined organic layer was washed with 100 mL of brine solution, dried and filtered with sodium sulfate, and concentrated under reduced pressure to obtain 4-benzylpyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxylic acid (1.25 g, 3.73 mmol, 44.9% yield) as a red solid.

[0091] Preparation Example 3: Preparation of 4-Benzyl-N-(3-fluoro-5-methylbenzyl)pyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0092]

[0093] HBTU (1.86 g, 4.91 mmol, 1.20 eq) was added at 0 °C to a solution of 4-benzylpyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxylic acid (1.25 g, 4.09 mmol, 1.00 eq) dissolved in DMA (30.0 mL). After 10 minutes, DIEA (1.16 g, 9.01 mmol, 2.20 eq) and (3-fluoro-5-methylphenyl)methaneamine (640 mg, 4.52 mmol, 1.10 eq) were added to the mixture. The mixture was stirred at 20 °C for 20 minutes. The reaction mixture was quenched at 25 °C by adding 100 mL of water, and extracted with ethyl acetate (50.0 mL, 3 times). The combined organic layer was washed with 100 mL of brine solution, dried with sodium sulfate, filtered, and concentrated under reduced pressure. 4-benzyl-N-(3-fluoro-5-methylbenzyl)pyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide (746 mg, 1.73 mmol, yield 42.3%) was produced as a yellow solid.

[0094] LC / MS: Rt = 0.476 min, m / z (M+H)+ = 427

[0095] [Example]

[0096] In the following Examples 1 to 26, compounds were synthesized using the same method as in the preparation example, or synthesized using appropriate reactants considering the structure of the compound to be synthesized. The structure of the compound synthesized in the present invention was confirmed by nuclear magnetic resonance (NMR) and mass spectrometry (MS).

[0097] Example 1: Synthesis of 4-Benzyl-N-(3-fluoro-5-methylbenzyl)pyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0098] 1H NMR: 400MHz, MeOD δ = 9.28 (s, 1H), 8.92 - 8.75 (m, 2H), 7.90 - 7.69 (m, 1H), 7.56 - 7.22 (m, 8H), 4.70 (s, 2H), 4.61 (d, 2H)

[0099] LC / MS: m / z (M+H)+ = 427

[0100] Example 2: Synthesis of 4-Benzyl-N-(3-fluoro-2-methoxybenzyl)pyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0101] 1H NMR: 400 MHz, MeOD δ = 8.81 (dd, 1H), 8.75 (dd, 1H), 8.20 - 8.12 (m, 1H), 7.68 (dd, 1H), 7.49 (d, 2H), 7.36 - 7.26 (m, 2H), 7.25 - 7.13 (m, 2H), 7.05 (br d, 2H), 4.73 (s, 2H), 4.60 (s, 2H), 3.99 (d, 3H)

[0102] LC / MS: m / z (M+H)+ = 443

[0103] Example 3: Synthesis of 4-Benzyl-N-(3,5-Difluoro-2-Methoxybenzyl)Pyrido[3',2':4,5]Furo[3,2-d]Pyrimidine-2-Carboxamide

[0104] 1H NMR: 400MHz, MeOD δ = 9.54 - 9.44 (m, 1H), 8.92 - 8.68 (m, 2H), 7.82 - 7.62 (m, 1H), 7.50 (d, 2H), 7.36 - 7.12 (m, 3H), 6.97 - 6.92 (m, 2H), 4.77 - 4.69 (m, 2H), 4.61 (s, 2H), 3.96 (d, 3H)

[0105] LC / MS: m / z (M+H)+ = 461.2

[0106] Example 4: Synthesis of 4-Benzyl-N-(2-methoxy-3-(trifluoromethyl)benzyl)pyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0107] 1H NMR: 400MHz, MeOD δ = 9.00 - 8.66 (m, 2H), 7.75 - 7.64 (m, 2H), 7.51 (d, 3H), 7.37 - 7.17 (m, 4H), 4.85 (s, 2H), 4.61 (s, 2H), 3.97 (s, 3H)

[0108] LC / MS: m / z (M+H)+ = 493

[0109] Example 5: Synthesis of 4-Benzyl-N-(2-chloro-6-methylbenzyl)pyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0110] 1H NMR: 400 MHz, MeOD δ = 8.90 - 8.70 (m, 3H), 7.71 (dd, J = 4.8, 7.6 Hz, 1H), 7.49 - 7.14 (m, 8H), 4.70 (d, J = 5.6 Hz, 2H), 4.53 (s, 2H), 2.48 (br s, 3H)

[0111] LC / MS: m / z (M+H)+ = 443

[0112] Example 6: Synthesis of 4-Benzyl-N-(5-methyl-2-(trifluoromethyl)benzyl)pyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0113] 1H NMR: 400 MHz, MeOD δ = 8.82 (dd, 1H), 8.75 (dd, 1H), 7.74 - 7.58 (m, 2H), 7.55 - 7.39 (m, 3H), 7.35 - 7.15 (m, 4H), 4.87 (br s, 2H), 4.60 (s, 2H), 2.37 (s, 3H)

[0114] LC / MS: m / z (M+H)+ = 477

[0115] Example 7: Synthesis of 4-Benzyl-N-(3-methoxy-4-methylbenzyl)pyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0116] 1H NMR: 400 MHz, MeOD δ = 8.87 - 8.66 (m, 2H), 7.67 (dd, 1H), 7.48 (d, 2H), 7.34 - 6.79 (m, 6H), 4.65 (s, 2H), 4.59 (s, 2H), 3.81 (s, 3H), 2.17 (s, 3H)

[0117] LC / MS: m / z (M+H)+ = 439.1

[0118] Example 8: Synthesis of N-(3-amino-5-(trifluoromethyl)benzyl)-4-benzylpyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0119] 1H NMR: 400 MHz, MeOD δ = 9.63 - 9.49 (m, 1H), 8.91 - 8.67 (m, 2H), 7.67 (dd, 1H), 7.49 (d, 2H), 7.38 - 7.13 (m, 3H), 7.34 - 6.81 (m, 2H), 4.70 - 4.51 (m, 4H)

[0120] LC / MS: m / z (M+H)+ = 478

[0121] Example 9: Synthesis of 4-Benzyl-N-(2-chloro-5-methylbenzyl)pyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0122] 1H NMR: 400 MHz, MeOD δ = 9.00 - 8.67 (m, 2H), 7.68 (dd, 1H), 7.51 (d, 2H), 7.30 - 7.00 (m, 6H), 4.79 - 4.72 (m, 2H), 4.61 (s, 2H), 2.30 (s, 3H)

[0123] LC / MS: m / z (M+H)+ = 443

[0124] Example 10: Synthesis of 4-Benzyl-N-(4-Methoxy-2-Methylbenzyl)Pyrido[3',2':4,5]Furo[3,2-d]Pyrimidine-2-Carboxamide

[0125] 1H NMR: 400 MHz, MeOD δ = 9.10 - 9.00 (m, 1H), 8.84 - 8.68 (m, 2H), 8.13 - 8.07 (m, 1H), 7.66 (dd, 1H), 7.46 (d, 2H), 7.34 - 7.13 (m, 4H), 6.82 - 6.64 (m, 2H), 4.64 - 4.59 (m, 2H), 4.57 (s, 2H), 3.77 (s, 3H)

[0126] LC / MS: m / z (M+H)+ = 439.2

[0127] Example 11: Synthesis of 4-Benzyl-N-(6-chloro-2-fluoro-3-methoxybenzyl)pyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0128] 1H NMR: 400MHz, MeOD δ = 9.15 - 8.99 (m, 1H), 8.91 - 8.58 (m, 2H), 7.67 (dd, 1H), 7.53 - 7.40 (m, 2H), 7.35 - 7.04 (m, 5H), 4.81 - 4.77 (m, 2H), 4.57 (s, 2H), 3.89 (s, 3H)

[0129] LC / MS: m / z (M+H)+ = 477

[0130] Example 12: Synthesis of 4-Benzyl-N-(3-chloro-4-(trifluoromethyl)benzyl)pyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0131] 1H NMR: 400 MHz, MeOD δ = 9.00 - 8.61 (m, 2H), 8.07 (s, 1H), 7.78 - 7.61 (m, 3H), 7.53 - 7.43 (m, 3H), 7.37 - 7.11 (m, 3H), 4.75 (s, 2H), 4.59 (s, 2H)

[0132] LC / MS: m / z (M+H)+ = 497

[0133] Example 13: Synthesis of N-(4-amino-3,5-dimethylbenzyl)-4-benzylpyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0134] 1H NMR: 400MHz, MeOD δ = 8.75 (s, 2H), 7.84 - 7.63 (m, 1H), 7.49 (d, 2H), 7.39 - 7.04 (m, 5H), 4.61 (d, 4H), 2.37 (s, 6H)

[0135] LC / MS: m / z (M+H)+ = 438.2

[0136] Example 14: Synthesis of 4-Benzyl-N-(3-hydroxy-2-methylbenzyl)pyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0137] 1H NMR: 400MHz, DMSO-d6 δ = 9.68(s, 1H), 9.26(s, 1H), 8.51(d, 1H), 8.43(d, 1H), 7.36(t, 1H), 7.28 - 7.20(m, 5H), 6.98(t, 1H), 6.91(d, 1H), 6.72(d, 1H), 4.23(s, 2H), 3.81(s, 2H), 2.08(s, 3H)

[0138] LC / MS: m / z (M+H)+ = 435.1

[0139] Example 15: Synthesis of 4-Benzyl-N-(2-fluoro-4-methoxybenzyl)pyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0140] 1H NMR: 400MHz, MeOD δ = 9.03 - 8.66 (m, 2H), 7.69 (dd, 1H), 7.50 (d, 2H), 7.42 - 7.13 (m, 4H), 6.86 - 6.61 (m, 2H), 4.72 - 4.66 (m, 2H), 4.60 (s, 2H), 3.81 (s, 3H)

[0141] LC / MS: m / z (M+H)+ = 443

[0142] Example 16: Synthesis of N-(2-amino-4-chlorobenzyl)-4-benzylpyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0143] 1H NMR: 400MHz, MeOD δ = 8.89 - 8.65 (m, 2H), 7.67 (dd, 1H), 7.48 (d, , 2H), 7.34 - 7.14 (m, 4H), 6.75 (d, 2H), 4.57 (d, 4H)

[0144] LC / MS: m / z (M+H)+ = 444.2

[0145] Example 17: Synthesis of 4-Benzyl-N-(2,4-Difluoro-3-Methoxybenzyl)Pyrido[3',2':4,5]Furo[3,2-d]Pyrimidine-2-Carboxamide

[0146] 1H NMR: 400 MHz, MeOD δ = 8.74 (s, 2H), 7.68 (dd, 1H), 7.49 (d, 2H), 7.34 - 6.90 (m, 5H), 4.71 (s, 2H), 4.60 (s, 2H), 3.97 (s, 3H)

[0147] LC / MS: m / z (M+H)+ = 461

[0148] Example 18: Synthesis of 4-Benzyl-N-(2-methyl-3-(trifluoromethyl)benzyl)pyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0149] 1H NMR: 400 MHz, MeOD δ = 9.60 - 9.38 (m, 1H), 8.94 - 8.59 (m, 2H), 7.82 - 7.56 (m, 3H), 7.49 (d, 2H), 7.38 - 7.13 (m, 4H), 4.81 - 4.73 (m, 2H), 4.60 (s, 2H), 2.51 (s, 3H)

[0150] LC / MS: m / z (M+H)+ = 477

[0151] Example 19: Synthesis of 4-Benzyl-N-(2-chloro-4-hydroxybenzyl)pyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0152] 1H NMR: 400MHz, MeOD δ = 9.20 (s, 1H), 8.81 (dd, 1H), 8.74 (dd, 1H), 7.67 (dd, 1H), 7.48 (d, 2H), 7.35 - 7.13 (m, 4H), 6.93 - 6.61 (m, 2H), 4.70 - 4.67 (m, 2H), 4.59 (s, 2H)

[0153] LC / MS: m / z (M+H)+ = 445

[0154] Example 20: Synthesis of 4-Benzyl-N-(2-chloro-4-hydroxybenzyl)pyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0155] 1H NMR: 400MHz, DMSO-d6 δ = 9.26(s, 1H), 9.0 (s, 1H), 8.51(d, 1H), 8.43(d, 1H), 7.36 - 7.18(m, 7H), 7.05(d, 1H), 6.59(d, 1H)

[0156] 4.23(s, 2H), 3.81(s, 2H),

[0157] LC / MS: m / z (M+H)+ = 445.1

[0158] Example 21: Synthesis of N-(2-chlorobenzyl)-4-(4-(trifluoromethyl)benzyl)pyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0159] 1H NMR: 400MHz, MeOD δ = 8.92 - 8.68 (m, 2H), 7.76 - 7.55 (m, 5H), 7.45 (dd, 2H), 7.34 - 7.25 (m, 2H), 4.79 (s, 2H), 4.71 (s, 2H)

[0160] LC / MS: m / z (M+H)+ = 497

[0161] Example 22: Synthesis of N-(2-methoxy-3-(trifluoromethyl)benzyl)-4-(4-(trifluoromethyl)benzyl)pyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0162] 1H NMR: 400MHz, DMSO-d6 δ = 9.26(s, 1H), 8.51(d, 1H), 8.43(d, 1H), 7.50(d, 2H), 7.44 - 7.17(m, 4H), 7.11(d, 2H), 4.23(s, 2H), 3.81(s, 2H), 3.78(s, 3H)

[0163] LC / MS: m / z (M+H)+ = 561

[0164] Example 23: Synthesis of N-(2-chloro-6-methylbenzyl)-4-(4-(trifluoromethyl)benzyl)pyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0165] 1H NMR: 400MHz, DMSO-d6 δ = 9.26(s, 1H), 8.51(d, 1H), 8.43(m, 1H), 7.58(d, 1H), 7.50(d, 2H), 7.36(t, 1H), 7.24 - 7.20(m, 2H), 7.11(d, 2H), 4.23(s, 2H), 3.81(s, 2H), 2.29(s, 3H)

[0166] LC / MS: m / z (M+H)+ = 511

[0167] Example 24: Synthesis of N-(2-chlorobenzyl)-4-(3-(trifluoromethyl)benzyl)pyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0168] 1H NMR: 400MHz, CDCl3 δ = 8.81 - 8.72 (m, 2 H) 8.50 (br t, J = 6.0 Hz, 1 H) 7.74 - 7.64 (m, 2 H) 7.62 - 7.38 (m, 5 H) 7.28 (br s, 1 H) 4.85 (d, 2 H) 4.63 (s, 2 H)

[0169] LC / MS: m / z (M+H)+ = 497.1

[0170] Example 25: Synthesis of N-(2-methoxy-3-(trifluoromethyl)benzyl)-4-(3-(trifluoromethyl)benzyl)pyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0171] 1H NMR: 400 MHz, DMSO-d6 δ = 9.26 (s, 1H), 8.51 (m, 1H), 8.43 (m, 1H), 7.47 - 7.17 (m, 8H), 4.23 (s, 2H), 3.81 (s, 2H), 3.78 (s, 3H)

[0172] LC / MS: m / z (M+H)+ = 561

[0173] Example 26: Synthesis of N-(2-chloro-6-methylbenzyl)-4-(3-(trifluoromethyl)benzyl)pyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0174] 1H NMR: 400MHz, DMSO-d6 δ = 9.26(s, 1H), 8.51(d, 1H), 8.43(d, 1H), 7.58(d, 1H), 7.47(d, 1H), 7.45 - 7.20(m, 6H), 4.23(s, 2H), 3.81(s, 2H), 2.29(s, 3H)

[0175] LC / MS: m / z (M+H)+ = 511

[0176]

[0177] [Test Example]

[0178] Test Example 1: Test of binding strength between KRAS G12D protein and a compound

[0179] To confirm the binding strength of the example compound to the KRAS G12D protein, the following experiment was conducted using an OCTET instrument.

[0180] The OCTET instrument is a biomolecular reaction analyzer capable of measuring intermolecular interactions in real time without separate fluorescent labels using BLI (Bio-Layer Interferometry) technology. Using this, the binding affinity between molecules was confirmed by measuring the phenomena of proteins and compounds binding and dissociating.

[0181] Recombinant KRAS G12D protein produced by Genomine Co., Ltd. was purchased and used. The recombinant KRAS G12D protein was biotinylated and used for OCTET binding affinity measurement. 200 μL of buffer (PBS + 1% DMSO + 0.02% Tween 20), ligand (1 / 10 KRAS G12D + Buffer), quench (EZ-Link + Buffer), and compounds of various concentrations were dispensed into each well of a Grainer black plate. Binding affinity was measured using the SSA sensor of the OCTET instrument in the sequence of Buffer-Loading (Ligand or Buffer)-Quenching-Buffer-Buffer (Baseline)-Compound-Buffer. K, with non-specific binding removed using the OCTET program D The value was derived.

[0182] MRTX1133 (CAS No.: 2621928-55-8) of Comparative Example 1 was purchased from MedChemExpress and used.

[0183] [Table 2]

[0184]

[0185] As shown in [Table 2] above, the compound according to the embodiment of the present invention specifically binds to the KRAS G12D protein, and it was confirmed that the binding strength to the KRAS G12D protein is higher compared to the substance of Comparative Example 1.

[0186]

[0187] Test Example 2: In vitro anticancer efficacy test against pancreatic cancer cell lines

[0188] IC utilizing pancreatic cancer cell lines AsPC-1 and HPAF-II with KRAS G12D mutations 50 The anticancer effects of the compounds in the examples were confirmed using an MTT assay. AsPC-1 and HPAF-II cell lines were purchased from the Korean Cell Line Bank (KCLB), and cells were cultured using RPMI1640 and MEM media. Cells were seeded at 3,000 cells / well in 96-well plates and cultured for 24 hours at 37°C in a 5% CO2 incubator. Each substance was dissolved in DMSO to prepare a stock solution, which was then diluted in the media to achieve a final DMSO concentration of 0.1% or less. After 24 hours of cell culture, the media was removed, and the cells were cultured for 3 days using the substances prepared at concentrations ranging from 1 nM to 100 uM. After 3 days, the medium was removed and washed with PBS, followed by the addition of 0.5 mg / ml MTT solution, and the cells were cultured for 3 hours at 37°C in a 5% CO2 incubator. Subsequently, the MTT solution was removed, the formazan crystals were dissolved in DMSO, and the absorbance was measured at 570 nm to determine the IC of the sample. 50 Values ​​were derived. The number of samples per sample was set to 3 or more.

[0189] [Table 3]

[0190]

[0191] As shown in [Table 3] above, the compounds of the embodiments of the present invention have an IC50 similar to or lower than that of Comparative Example 1 in KRAS G12D mutant pancreatic cancer cell lines. 50 It was confirmed that it has a value. The compound of the embodiment of the present invention specifically binds to the KRAS G12D protein and was confirmed to have excellent anticancer activity by inhibiting cell growth in related pancreatic cancer cell lines.

Claims

1. Compounds, solvates, stereoisomers, or pharmaceutically acceptable salts of the following chemical formula I: [Chemical Formula I] In the above formula, R1 and R2 are each independently H or -CF3, and R3 to R7 are each independently H, F, Cl, -CF3, -NH2, -OH, -C 1-6 alkyl or -OC 1-6 It is an alkyl.

2. In Claim 1, Compounds of Formula I, solvates, stereoisomers, or pharmaceutically acceptable salts thereof, selected from the group consisting of compounds represented by the following chemical formulas: .

3. A pharmaceutical composition for the prevention or treatment of cancer, comprising as an active ingredient a compound, solvate, stereoisomer, or pharmaceutically acceptable salt thereof according to any one of claims 1 to 2.

4. In Claim 3, A pharmaceutical composition for the prevention or treatment of cancer, wherein the cancer comprises tumor tissue in which a KRAS G12D mutation is present.

5. In Claim 3, A pharmaceutical composition for the prevention or treatment of cancer, wherein the cancer is any one selected from the group consisting of breast cancer, lung cancer, stomach cancer, prostate cancer, uterine cancer, ovarian cancer, kidney cancer, pancreatic cancer, liver cancer, colorectal cancer, colon cancer, skin cancer, head and neck cancer, melanoma, and thyroid cancer.

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