Novel compound for treating cancer in which KRAS g12d mutation is present, and composition for preventing or treating cancer using same

Abstract

The present invention relates to a novel compound specifically binding to the KRAS G12D protein, and a pharmaceutical composition, comprising same, for preventing or treating cancer in which the KRAS G12D mutation is present.

Description

Novel compound for treating cancer with KRAS G12D mutation and composition for preventing or treating 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 comprising the same for the prevention or treatment of cancer in which a KRAS G12D mutation exists. Specifically, the novel compound that specifically binds to the KRAS G12D protein of the present invention has excellent binding affinity to the KRAS G12D protein and has a cell growth inhibitory activity effect in cancer in which a KRAS G12D mutation exists, and thus a pharmaceutical composition using the same for the prevention or treatment of cancer can be provided.

[0002] The MAPK / ERK signaling pathway regulates various cellular responses, including cell proliferation, differentiation, and apoptosis, by transducing extracellular stimuli to the nucleus. The KRAS protein is an initiator of the MAPK / ERK signaling pathway and functions as a switch responsible for inducing cell division. In the inactive state of the V-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS), the KRAS protein binds to guanosine diphosphate (GDP), effectively transmitting a negative signal to inhibit cell division. In response to extracellular signals, the KRAS protein is allosterically activated, allowing the nucleotide exchange of GDP for guanosine triphosphate (GTP). In the GTP-bound active state of the KRAS protein, KRAS recruits and activates other cell signaling receptors as well as proteins required for the propagation of growth factor-induced signaling. Examples of proteins recruited by KRAS-GTP include 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 suppress cell division. KRAS gain-of-function mutations exhibit increased GTP binding and a reduced ability to convert GTP to GDP. The result is increased MAPK / ERK signaling, which promotes cancer cell growth. In other words, when an amino acid substitution occurs due to a mutation in the KRAS gene, KRAS is permanently activated due to a decreased function as a GTPase or a decreased response to GTPase-activating proteins, continuously transmitting signals downstream. This excessive signaling leads to carcinogenesis and enhanced cancer proliferation.

[0003] The KRAS gene is one of several genes implicated in the development of non-small cell lung cancer, colon cancer, and pancreatic cancer, and KRAS mutations are the most common cancer-causing factors. It is known that approximately 25% of all cancers, 30% of lung adenocarcinoma and colon cancer, and 80% of pancreatic cancer harbor KRAS mutations. The most commonly targeted mutations are KRAS G12C, KRAS G12V, and KRAS G12D.

[0004] Pancreatic cancer, of which ductal adenocarcinoma is the mainstay, has 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 worldwide each year (CA Cancer J. Clin., 2018, 68, p. 394-424). The most effective treatment for pancreatic cancer is surgery, but early detection is difficult, so it often metastasizes, making surgery ineffective. If surgery is not the treatment, chemotherapy or radiation therapy are available, but the survival rate is poor. Currently, FOLFRINOX therapy (multi-drug combination therapy with levofolinate added to three types of chemotherapy drugs: 5-FU, irinotecan, and oxaliplatin) is used as the standard treatment for pancreatic cancer, but because of its high toxicity, careful patient selection is necessary, such as limiting the patients to whom it is prescribed to have an ECOG Performance Status of 1 or lower (J. Clin. Oncol., 2018, 36, p. 2545-2556). As a molecularly targeted therapy, the epidermal growth factor receptor (EGFR) inhibitor erlotinib has been approved in combination with gemcitabine, but the extension of overall survival is only about 2 weeks compared to gemcitabine alone, and satisfactory therapeutic effects have not been obtained, so highly effective treatments are still needed (J. Clin. Oncol., 2007, 25, p. 1960-1966).

[0005] Pancreatic ductal adenocarcinoma, the most common type of pancreatic cancer, is thought to develop from mild to severe stages of pancreatic intraepithelial neoplasia (PanIN). KRAS gene mutations are already present in early-stage PanIN. Subsequently, abnormalities in tumor suppressor genes such as INK4A, p53, or SMAD4 develop, leading to malignant transformation (Nature Rev. Cancer, 2010, 10, p. 683-695). Furthermore, mutations in the KRAS gene are found in more than 90% of pancreatic ductal adenocarcinomas, with point mutations in codon 12, located in KRAS exon 2, accounting for the majority (Cancer Cell, 2017, 32, p. 185-203). This suggests that KRAS plays a crucial role in the carcinogenesis and development of pancreatic cancer.

[0006] Against this backdrop, the inventors of the present invention have completed the present invention by identifying a novel compound that specifically binds to the KRAS G12D protein, exhibiting excellent binding affinity for the KRAS G12D protein and demonstrating cell growth inhibition activity against cancer cells harboring KRAS G12D mutations. In particular, the inventors have confirmed that the compound exhibits excellent growth inhibition activity against pancreatic cancer cells and can address the issues of disease recurrence and resistance to therapeutic agents.

[0007] [Prior Art Literature]

[0008] [Patent Document]

[0009] Domestic Publication Patent 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 preventing or treating cancer, which comprises the compound as an active ingredient, having excellent binding affinity for the KRAS G12D protein and excellent cell growth inhibitory activity in cancer in which a KRAS G12D mutation exists.

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

[0012] [Chemical Formula I]

[0013]

[0014] In the above formula,

[0015] X is O or S,

[0016] R1 and R2 are each independently H or R1 and R2 are combined with each other to form a 6-membered ring,

[0017] R3 is C 1-6 Alkyl, phenyl, C 3-6 Cycloalkyl, C 3-6 Heterocycloalkyl or C 3-6 It is heteroaryl,

[0018] C above 1-6 Alkyl, phenyl, C 3-6 Cycloalkyl, C 3-6 Heterocycloalkyl or C 3-6 At least one hydrogen atom of the heteroaryl is substituted with one or more substituents selected from the group consisting of halogen and carbon.

[0019] In addition, the present invention provides a pharmaceutical composition for preventing or treating cancer, comprising the compound, solvate, stereoisomer or pharmaceutically acceptable salt thereof as an active ingredient.

[0020] The compound of formula I, a solvate, a stereoisomer, or a pharmaceutically acceptable salt thereof according to the present invention is a novel compound that specifically binds to the KRAS G12D protein, has excellent binding affinity to the KRAS G12D protein, has a cell growth inhibitory effect on cancer cells harboring a KRAS G12D mutation, and can be used for the prevention or treatment of various cancers. A pharmaceutical composition comprising the same can be useful for the prevention or treatment of cancer, and can be particularly useful for the prevention or treatment of pancreatic cancer.

[0021] Hereinafter, various embodiments of the present invention will be described with reference to the accompanying drawings. It should be understood that the present invention is not limited to specific embodiments, but rather encompasses 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.

[0022] In this document, the expressions “has”, “may have”, “includes”, or “may include” indicate the presence of a feature (e.g., a number, function, operation, or component such as a part), but do not exclude the presence of additional features.

[0023] In this document, the expressions "A or B," "at least one of A and / or B," or "one or more of A and / or B" can include all possible combinations of the listed items. For example, "A or B," "at least one of A and B," or "at least one of A or B" can all refer to cases where (1) at least one A is included, (2) at least one B is included, or (3) at least one A and at least one B are included.

[0024] The expression "configured to" as used herein can be used interchangeably 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".

[0025] The terms used in this document are used only to describe specific embodiments and may not be intended to limit the scope of other embodiments. The singular expression may include the plural expression unless the context clearly indicates otherwise. Terms used herein, including technical or scientific terms, may have the same meaning as commonly understood by those of ordinary skill in the art described in this document. Terms defined in general dictionaries among the terms used in this document may be interpreted as having the same or similar meaning in the context of the relevant technology, and shall not be interpreted in an idealized or overly formal sense unless explicitly defined in this document. In some cases, even if a term is defined in this document, it cannot be interpreted to exclude the embodiments of this document.

[0026] 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 present invention. Therefore, the scope of this document should be interpreted to include all modifications or various other embodiments based on the technical concept of the present invention.

[0027] Hereinafter, preferred embodiments of the present invention will be described in detail. Prior to this, it should be noted that the terms and words used in this specification and claims should not be construed as limited to their conventional or dictionary meanings. Based on the principle that the inventor can appropriately define the concepts of terms to best explain his or her invention, they should be interpreted in a way that aligns with the technical spirit of the present invention.

[0028] Accordingly, the configurations of the embodiments described in this specification are only some of the most preferred embodiments of the present invention and do not represent all of the technical ideas of the present invention, so it should be understood that there may be various equivalents and modified examples that can replace them at the time of filing this application.

[0029] Throughout the specification, whenever a part is said to "include" a component, this does not mean that it excludes other components, but rather that it may include other components, unless otherwise stated.

[0030] Hereinafter, the present invention will be described in detail.

[0031]

[0032] The present invention relates to a compound represented by Chemical Formula I, and more particularly, to a novel compound that specifically binds to KRAS G12D protein and a pharmaceutical composition for preventing or treating cancer comprising the same as an active ingredient.

[0033] [Chemical Formula I]

[0034]

[0035] Unless otherwise stated, terms used in the description and claims of the present invention have the meanings set forth below.

[0036] According to the custom used in the art, in the chemical formula of this application " " is used to indicate that a residue or substituent "R" is attached to the skeletal structure.

[0037] "Alkyl" is a hydrocarbon group having primary, secondary, tertiary and / or quaternary carbon atoms, including saturated aliphatic groups which may be straight-chain, branched or cyclic, or a combination thereof. For example, an alkyl group may have from 1 to 20 carbon atoms (i.e., C1-C 20 alkyl), 1 to 10 carbon atoms (i.e., C1-C 10alkyl), or may have 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), 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), 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).

[0038] Moreover, the term "alkyl" as used throughout the specification, examples and claims is intended to encompass both unsubstituted and substituted alkyl groups, the latter of which refers to alkyl moieties having substituents replacing hydrogens on one or more carbons of the hydrocarbon backbone, including haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl.

[0039] The term "C x-y " or "C x -C y "When used with chemical moieties such as acyl, acyloxy, alkyl, alkenyl, alkynyl or alkoxy, it is considered to include groups containing x to y carbons in the chain. C0 alkyl 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.

[0040] "Alkoxy" refers to a group having the formula -O-alkyl, wherein an alkyl group as defined above is attached to the parent compound through an oxygen atom. The alkyl moiety of an alkoxy group may be, for example, a group having from 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 alkoxy), or may have 1 to 6 carbon atoms (i.e., C1-C6alkoxy). 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).

[0041] "Alkenyl" means a group having primary, secondary, tertiary and / or quaternary carbon atoms, including straight-chain, branched and cyclic groups, or combinations thereof, and having one or more regions of unsaturation, i.e., carbon-carbon sp 2A hydrocarbon with a double bond. For example, an alkenyl group has 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 alkenyl), or may have 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).

[0042] "Alkynyl" is a hydrocarbon having primary, secondary, tertiary and / or quaternary carbon atoms, including straight-chain, branched and cyclic groups, or combinations thereof, and having at least one carbon-carbon sp triple bond. For example, an alkynyl group has from 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 alkynyl), or may have 2 to 6 carbon atoms (i.e., C2-C6alkynyl). Examples of suitable alkynyl groups include, but are not limited to, acetylenyl (-C≡CH) and propynyl (-CH2C≡CH).

[0043] As used herein, the term "aryl" includes substituted or unsubstituted monovalent or divalent aromatic hydrocarbon groups, which are monocyclic, bicyclic, or polycyclic, each ring atom being 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. An 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 at least one of the rings is aromatic, for example, the other cyclic ring may be a cycloalkyl, a cycloalkenyl, a cycloalkynyl, an aryl, a heteroaryl, and / or a heterocycloalkyl. Aryl groups include benzene, naphthalene, phenanthrene, anthracene, indene, indane, phenol, aniline, and the like.

[0044] The term "carbocyclylalkyl", or "cycloalkylalkyl", or "(cycloalkyl)alkyl" as used herein refers to an alkyl group substituted with a carbocycle group or a cycloalkyl group.

[0045] The terms "carbocycle," "carbocyclyl," "carbocyclic," or "cycloalkyl," as used herein, refer to a non-aromatic, saturated or unsaturated, monovalent or divalent ring, which may be monocyclic, bicyclic, or polycyclic, and wherein each ring atom is carbon. A cycloalkyl group may have 3 to 7 carbon atoms as a monocycle, 7 to 12 carbon atoms as a bicycle, and up to about 20 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 exemplary cycloalkyl groups, 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 cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like. Unless otherwise specified, the cycloalkyl may be substituted with one or more substituents described herein.

[0046] The terms “heterocyclylalkyl” and “heterocycloalkyl” as used herein refer to an alkyl group substituted with a heterocycloalkyl group.

[0047] The terms "heterocyclyl", "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, wherein the ring structure comprises one or more heteroatoms, preferably 1 to 4 heteroatoms, more preferably 1 to 2 heteroatoms. The terms "heterocyclyl," "heterocycle," "heterocyclic," and "heterocycloalkyl" also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings, wherein at least one of the rings is heterocyclic, for example, the other cyclic ring can be a cycloalkyl, a cycloalkenyl, a cycloalkynyl, an aryl, a heteroaryl, and / or a heterocyclyl. Bicyclic and polycyclic heterocyclic ring systems can be fused, bridged, or spiro ring systems. Substituted heterocycles include, for example, heterocyclic rings substituted with any of the substituents disclosed herein, including a carbonyl group. Heterocyclic groups include, for example, piperidine, piperazine, pyrrolidine, morpholine, lactones, lactams, and the like.Additionally, exemplary heterocycles include dihydropyridyl, dihydroindolyl, tetrahydropyridyl (piperidyl), tetrahydrothiophenyl, sulfur-oxidized tetrahydrothiophenyl, indolenyl, piperidinyl, 4-piperidinyl, pyrrolidinyl, 2-pyrrolidonyl, pyrrolinyl, tetrahydrofuranyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, octahydroisoquinolinyl, 6H-1,2,5-thiadiazinyl, 2H,6H-1,5,2-dithiazinyl, pyranyl, chromenyl, xanthenyl, phenoxathinyl, 2H-pyrrolyl, 3H-indolyl, 4H-quinolizinyl, phthalazinyl, naphthyridinyl, quinoxalinyl, Examples include, but are not limited to, quinazolinyl, cinnolinyl, pteridinyl, 4aH-carbazolyl, carbazolyl, B-carbolinyl, phenanthridinyl, acridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, furazanyl, phenoxazinyl, isochromanyl, chromanyl, imidazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolinyl, piperazinyl, methyl piperazinyl, quinuclidinyl, morpholinyl, azabicyclo(2.1.1)hexanyl, azacycloheptanyl, 1-oxa-3-aza-cycloheptanyl, azetidinyl, aziridinyl, and oxazolidinyl (each of which may be substituted or unsubstituted).

[0048] "Heteroaryl" refers to a substituted or unsubstituted monovalent or divalent aromatic group, which is monocyclic, bicyclic or polycyclic, containing one or more heteroatoms in the ring. Non-limiting examples of suitable heteroatoms that can 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 at least one of the rings is heteroaromatic, for example, the other cyclic ring can be a cycloalkyl, a cycloalkenyl, a cycloalkynyl, an aryl, a heteroaryl, and / or a heterocyclyl. Heterocyclic groups include, for example, benzofuran, benzothiophene, pyrrole, furan, thiophene, imidazole, indole, isoindole, isoxazole, isothiazole, oxazole, thiazole, quinoline, isoquinoline, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine, each of which may be substituted or unsubstituted.

[0049] As used herein, the terms “halo” and “halogen” mean halogen, including chloro, fluoro, bromo, and iodine.

[0050] The present invention relates to a compound of the following formula I, a solvate, a stereoisomer or a pharmaceutically acceptable salt thereof.

[0051] [Chemical Formula I]

[0052]

[0053] In the above formula,

[0054] X is O or S,

[0055] R1 and R2 are each independently H or R1 and R2 are combined with each other to form a 6-membered ring,

[0056] R3 is C 1-6 Alkyl, phenyl, C 3-6 Cycloalkyl, C 3-6 Heterocycloalkyl or C3-6 It is heteroaryl,

[0057] C above 1-6 Alkyl, phenyl, C 3-6 Cycloalkyl, C 3-6 Heterocycloalkyl or C 3-6 At least one hydrogen atom of the heteroaryl is substituted with one or more substituents selected from the group consisting of halogen and carbon.

[0058] Specifically, in the present invention, the 6-membered ring formed by combining R1 and R2 may be selected from the group consisting of structural formulas 1 to 8 of the following [Table 1].

[0059] [Table 1]

[0060]

[0061]

[0062] Specifically, in the present invention, the C 1-6 Alkyl is any one selected from the group consisting of ethyl and isopropyl,

[0063] C above 3-6 Cycloalkyl is cyclohexyl,

[0064] C above 3-6 Heterocycloalkyl is morpholinyl,

[0065] C above 3-6 Heteroaryl may be any one selected from the group consisting of pyridinyl and furanyl.

[0066] Specifically, in the present invention, the compound of formula I, a solvate, a stereoisomer or a pharmaceutically acceptable salt thereof selected from the group consisting of compounds represented by the chemical formulas of [Table 2] below, but not limited thereto.

[0067] [Table 2]

[0068]

[0069]

[0070]

[0071]

[0072]

[0073]

[0074] In addition, the present invention specifically relates to a pharmaceutical composition for preventing or treating cancer, comprising a compound of the above chemical formula I, a solvate, a stereoisomer or a pharmaceutically acceptable salt thereof as an active ingredient.

[0075] The term "cancer" is a general term for diseases caused by cells that have aggressive properties, such as dividing and multiplying in violation of normal growth limits, invasive properties, such as infiltrating surrounding tissues, and metastatic properties, such as spreading to other parts of the body.

[0076] The above cancer may be a cancer that includes tumor tissue in which a KRAS G12D mutation exists.

[0077] Mutations in codons 12 and 13 of the KRAS gene result in functional changes in the p21-ras protein, which is the product of this gene, and as a result, it promotes cell growth and division by transmitting growth signals to the cell nucleus excessively, thereby participating in the carcinogenesis process. KRAS mutations can appear in various ways depending on the change in location and sequence, and representative mutations include G12D, G12V, G12R, G12C, in which the amino acid residue at position 12, glycine, is substituted with aspartic acid, valine, arginine, or cysteine, or G13D, G13V, G13H, in which the amino acid residue at position 13, glycine, is substituted with aspartic acid, valine, or histidine. Depending on the type of KRAS mutation, the oncological appearance varies, and the effectiveness of anticancer treatments such as cetuximab or panitumab can be determined, and it has been reported that this results in differences in patient survival rates. KRAS mutations are very common in human cancers, occurring in approximately 90% of pancreatic cancers, 50% of colorectal cancers, and 30% of non-small cell lung cancers, and most of these mutations are known to be concentrated in codons 12 and 13.

[0078] The cancer may be at least 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, colon cancer, colon cancer, skin cancer, head and neck cancer, melanoma, and thyroid cancer, but is not limited thereto. Preferably, the cancer may be pancreatic cancer, but is not limited thereto.

[0079] In addition, from the experimental results of the examples, the compound of chemical formula I of the present invention has an effect of inhibiting the growth of cancer cells including tumor tissues in which KRAS G12D mutation exists, and can exhibit excellent therapeutic effects on various cancer-related diseases.

[0080] As used herein, the term "treating" or "treatment" means inhibiting a disease, condition or disorder, e.g., inhibiting the disease, condition or disorder in a subject experiencing or exhibiting the pathology or signs of the disease, condition or disorder, i.e., preventing further development of the pathology and / or signs, or ameliorating a disease, condition or disorder, e.g., ameliorating the disease, condition or disorder in a subject experiencing or exhibiting the pathology or signs of the disease, condition or disorder, i.e., reversing the pathology and / or signs, e.g., reducing the severity of the disease.

[0081] As used herein, the term "preventing" or "prevention" refers to preventing a disease, condition or disorder, for example, in a subject who may be predisposed to the disease, condition or disorder but does not yet experience or exhibit the pathology or signs of the disease.

[0082] In the present invention, the pharmaceutical composition may include a conventional pharmaceutically acceptable carrier, excipient, or additive. The pharmaceutical composition may be formulated according to a conventional method, and may be prepared in various oral administration forms such as tablets, pills, powders, capsules, syrups, emulsions, and microemulsions, or in parenteral administration forms such as intramuscular, intravenous, or subcutaneous administration.

[0083] When the pharmaceutical composition of the present invention is manufactured in the form of an oral dosage form, 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 manufactured in the form of an injection, examples of additives or carriers include water, saline solution, glucose aqueous solution, pseudo-saccharide aqueous solution, alcohol, glycol, ether (e.g., polyethylene glycol 400), oil, fatty acid, fatty acid ester, glyceride, surfactant, suspending agent, emulsifier, etc.

[0084] The dosage of the pharmaceutical composition is an amount effective for the treatment or prevention of an individual or patient, and may be administered orally or parenterally depending on the purpose. In the case of oral administration, the amount is 0.001 to 10 mg, more specifically 0.1 to 10 mg, per kg of body weight per day based on the active ingredient. In the case of parenteral administration, the amount is 0.01 to 10 mg, more specifically 0.1 to 10 mg, per kg of body weight per day based on the active ingredient. It should be understood that the dosage for a specific individual or patient should be determined in light of various related factors such as the patient's weight, age, sex, health condition, diet, administration time, administration method, and severity of the disease, and may be appropriately increased or decreased by a specialist. The dosage is not intended to limit the scope of the present invention in any way. A doctor or veterinarian having ordinary skill in the relevant art can easily determine and prescribe an effective amount of the required pharmaceutical composition. For example, a physician or veterinarian may start the dosage of a 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.

[0085] Additionally, the pharmaceutical composition may be administered for oncology therapy in combination with chemotherapy, radiation therapy, immunotherapy, hormonal therapy, bone marrow transplantation, stem cell replacement therapy, other biological therapies, surgical intervention, or a combination thereof. For example, it may be used as adjuvant therapy alongside other long-term treatment strategies, or to maintain the patient's condition after tumor regression or chemoprevention in critically ill patients.

[0086] Hereinafter, the present invention will be described in more detail through examples. These examples are intended solely to illustrate the present invention more specifically, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples, in accordance with the gist of the present invention.

[0087]

[0088] [Manufacturing example]

[0089] Compounds 1 to 32 of the present invention were prepared according to the following manufacturing examples and examples.

[0090] Manufacturing Example 1: Preparation of 4-benzylfuro[3,2-d]pyrimidine-2-carboxylic acid

[0091]

[0092] Manufacturing Example 1-1: Manufacturing of ethyl 4-chlorofuro[3,2-d]pyrimidine-2-carboxylate

[0093] Ethyl 4-hydroxyfuro[3,2-d]pyrimidine-2-carboxylate (2.0 g, 9.6 mmol, 1.0 eq) and phosphoryl trichloride (14.8 g, 96.6 mmol, 9.00 ml, 10.1 eq) were mixed and reacted at 80 °C for 5 hours. After completion of the reaction, the mixture was concentrated under reduced pressure and dissolved in toluene (25 ml). The mixture was concentrated under reduced pressure to remove toluene, and ethyl 4-chlorofuro[3,2-d]pyrimidine-2-carboxylate (2.1 g, 9.3 mmol, 96.8% yield) was obtained.

[0094] 1H-NMR (400MHz, DMSO-d6) δ 8.13(d, 1H), 6.30(d, 1H), 4.49(m, 2H), 1.38(t, 3H)

[0095] LC / MS [M+H]+ 226.01

[0096] Manufacturing Example 1-2: Preparation of ethyl 4-benzylfuro[3,2-d]pyrimidine-2-carboxylate

[0097] Ethyl 4-chlorofuro[3,2-d]pyrimidine-2-carboxylate (500 mg, 2.2 mmol, 1.0 eq), 2-benzyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (650 mg, 3.0 mmol, 1.35 eq), potassium carbonate (600 mg, 4.3 mmol, 2.0 eq), and Pd(dppf)Cl2 (150 mg, 205 umol) are dissolved in water (2.0 ml) and dioxane (5.0 ml), and mixed under vacuum and nitrogen atmosphere at 20 °C. The mixture is reacted at 50 °C for 12 hours. After the reaction, filter and concentrate the filtrate. The concentrated product was purified by column to obtain ethyl 4-benzylfuro[3,2-d]pyrimidine-2-carboxylate (52 mg, 184 μmol, 8.4% yield).

[0098] 1H-NMR (400MHz, DMSO-d6) δ 8.13(d, 1H), 7.28-7.20(m, 5H), 6.30(d, 1H), 4.49(m, 2H), 3.81(s, 2H), 1.38(t, 3H)

[0099] LC / MS [M+H]+ 282.10

[0100] Manufacturing Example 1-3: Manufacturing of 4-benzylfuro[3,2-d]pyrimidine-2-carboxylic acid

[0101] Ethyl 4-benzylfuro[3,2-d]pyrimidine-2-carboxylate (50 mg, 177 μmol, 1.0 eq) and LiOH.H2O (21.0 mg, 500 μmol, 2.8 eq) were mixed with methanol (0.5 ml), water (0.5 ml), and tetrahydrofuran (0.5 ml) at 20 °C under vacuum and nitrogen atmosphere and reacted for 1 hour. After the reaction, 1 M aqueous hydrochloric acid solution (5.0 ml) was added and extracted with ethyl acetate (3.0 ml, 3 times). The organic layer was dried over anhydrous magnesium sulfate, filtered, and the filtrate was distilled under reduced pressure to obtain the product 4-benzylfuro[3,2-d]pyrimidine-2-carboxylic acid (38 mg, 149 μmol, 84.2% yield).

[0102] 1H-NMR (400MHz, DMSO-d6) δ 12.7(s, 1H), 8.13(d, 1H), 7.28-7.20(m, 5H), 6.30(d, 1H), 3.81(s, 2H)

[0103] LC / MS [M+H]+ 254.07

[0104] Manufacturing Example 2: Preparation of 4-benzyl-N-propylfuro[3,2-d]pyrimidine-2-carboxamide

[0105]

[0106] 4-Benzylfuro[3,2-d]pyrimidine-2-carboxylic acid (35.0 mg, 138 μmol, 1.0 eq) and propanamine (12.2 mg, 207 μmol, 1.5 eq) were dissolved in dimethylformamide (3.0 ml), and HATU (36.2 mg, 276 μmol, 2.0 eq) and DIEA (26.0 mg, 183 μmol, 2.6 eq) were added dropwise at 20 °C, followed by reaction at 25 °C for 1 hour. The reaction mixture was concentrated under reduced pressure and purified by column chromatography to obtain 4-benzyl-N-propylfuro[3,2-d]pyrimidine-2-carboxamide (21 mg, 71 μmol, 51.4% yield).

[0107] 1H-NMR (400MHz, DMSO-d6) δ 8.93(s, 1H), 8.13(d, 1H), 7.28-7.20(m, 5H), 6.30(d, 1H), 3.81(s, 2H), 3.25(t, 2H), 1.55(m, 2H), 0.88(t, 3H)

[0108] LC / MS [M+H]+ 296.14

[0109] Manufacturing Example 3: Manufacturing of ethyl 4-hydroxybenzofuro[3,2-d]pyrimidine-2-carboxylate

[0110]

[0111] Manufacturing Example 3-1: Manufacturing of methyl 3-aminobenzofuran-2-carboxylate

[0112] 2-Chlorobenzonitrile (29.8 g, 216.5 mmol, 1.00 eq) was first dissolved in dimethylformamide (300 ml), and methyl 2-hydroxyacetate (23.5 g, 260.9 mmol, 1.20 eq) and cesium carbonate (140.0 g, 429.7 mmol, 1.98 eq) were added dropwise at 20 °C, and the mixture was stirred at 80 °C for 2 hours. When the reaction was complete, water (1500 ml) was added at 25 °C to terminate the reaction, and the product was extracted with ethyl acetate (500 ml, 3 times). The organic layer was dried over anhydrous magnesium sulfate, filtered, and the filtrate was distilled under reduced pressure to obtain methyl 3-aminobenzofuran-2-carboxylate (21.4 g, 111.9 mmol, 51.7% yield).

[0113] 1H-NMR (400MHz, DMSO-d6) δ 7.84(d, 1H), 7.59(d, 1H), 7.39(t, 1H), 7.31(t, 1H), 5.82(s, 2H), 3.89(s, 3H)

[0114] LC / MS [M+H]+ 191.06

[0115] Manufacturing Example 3-2: Manufacturing of 3-aminobenzofuran-2-carboxamide

[0116] Methyl 3-aminobenzofuran-2-carboxylate (21.4 g, 111.9 mmol, 1.00 eq) and ammonia / methanol (7 M, 144 mL) were added dropwise to a Teflon tank at 20 °C and stirred at 80 °C for 12 hours. After the reaction was completed, the product was obtained by concentration under reduced pressure. The product was purified using a silica column to obtain 3-aminobenzofuran-2-carboxamide (10.3 g, 58.5 mmol, 52.3% yield).

[0117] 1H-NMR (400MHz, DMSO-d6) δ 10.4(s, 1H), 7.84(d, 1H), 7.59(d, 1H), 7.39(t, 1H), 7.31(t, 1H), 6.98(s, 2H), 4.17(m, 2H), 1.24(t, 3H)

[0118] LC / MS [M+H]+ 176.06

[0119] Manufacturing Example 3-3: Manufacturing of ethyl 2-((2-carbamoylbenzofuran-3-yl)amino)-2-oxoacetate

[0120] 3-Aminobenzofuran-2-carboxamide (10.3 g, 58.5 mmol, 1.00 eq) and pyridine (13.94 g, 175.5 mmol, 3.00 eq) were dissolved in tetrahydrofuran (104.2 ml), and ethyl 2-chloro-2-oxoacetate (24.12 g, 175.5 mmol, 3.00 eq) was added dropwise at 0 °C, followed by reaction at 20 °C for 1 hour. The reaction product was filtered and purified to obtain ethyl 2-((2-carbamoylbenzofuran-3-yl)amino)-2-oxoacetate (6.2 g, crude).

[0121] 1H-NMR (400MHz, DMSO-d6) δ 11.40(s, 1H), 7.70(d, 1H), 7.65(d, 1H), 7.36(t, 1H), 7.22(s, 1H), 4.49(m, 2H), 1.38(t, 3H)

[0122] LC / MS [M+H]+ 276.07

[0123] Manufacturing Example 3-4: Manufacturing of ethyl 4-hydroxybenzofuro[3,2-d]pyrimidine-2-carboxylate

[0124] Ethyl 2-((2-carbamoylbenzofuran-3-yl)amino)-2-oxoacetate (6.2 g, 10.7 mmol, 48% purity, 1.00 eq) is dissolved in ethanol (75.0 ml). Sodium methoxide (12.7 g, 37.3 mmol, 20% purity, 3.50 eq) is added dropwise at 45 °C and stirred for 12 h. The reaction mixture is filtered and the filtrate is concentrated. The concentrate is dissolved in dichloromethane (50 ml) and washed with water (50 ml). The aqueous layer is adjusted to pH 2 with 1 M hydrochloric acid aqueous solution. The organic layer is obtained using dichloromethane (50 ml, 3 times). The organic layer was dried over anhydrous magnesium sulfate, filtered, and the filtrate was distilled under reduced pressure to obtain ethyl 4-hydroxybenzofuro[3,2-d]pyrimidine-2-carboxylate (810 mg, 3.1 mmol, 29.0% yield).

[0125] 1H-NMR (400MHz, CDCl3) δ 8.7 (s, 1H), 7.42 (d, 1H), 7.28-7.20 (m, 5H), 6.30 (d, 1H), 3.81 (s, 2H), 3.25 (t, 2H), 1.55 (m, 2H), 0.88(t, 3H)

[0126] LC / MS [M+H]+ 258.06

[0127]

[0128] In the following Examples 1 to 32, the compounds were synthesized in the same manner as in the Preparation Examples, or synthesized using appropriate reactants taking into account the structure of the compounds to be synthesized. The structures of the compounds synthesized in the present invention were confirmed by nuclear magnetic resonance (NMR) and mass spectrometry (MS). NMR was measured with a Bruker Avance 300 instrument. The solvents for measurement were deuterated chloroform (CDCl3) or deuterated dimethyl sulfoxide (DMSO-d6), and the internal standard was tetramethylsilane (TMS). Mass spectrometry was measured with a Waters ACQUITY Qda instrument, and analyzed in electrospray ionization (ESI) positive ion mode. The known starting materials of the present invention can be prepared by conventional synthetic methods in the art, or can be purchased from Sigma-Aldrich, TCI, Wako, Kanto, Fluorchem, Acros, Alfa, Fluka, Combi-Blocks, Dae-Jung, etc.

[0129]

[0130] Example 1: Synthesis of 4-benzyl-N-propylfuro[3,2-d]pyrimidine-2-carboxamide

[0131] 1H-NMR (400MHz, DMSO-d6) δ 8.93(s, 1H), 8.13(d, 1H), 7.28-7.20(m, 5H), 6.30(d, 1H), 3.81(s, 2H), 3.25(t, 2H), 1.55(m, 2H), 0.88(t, 3H)

[0132] LC / MS [M+H]+ 296.25

[0133] Example 2: Synthesis of 4-benzyl-N-isobutylfuro[3,2-d]pyrimidine-2-carboxamide

[0134] 1H-NMR (400MHz, DMSO-d6) δ 8.98(s, 1H), 8.13(d, 1H), 7.28-7.20(m, 5H), 6.30(d, 1H), 3.81(s, 2H), 2.88(d, 2H), 2.34(m, 1H), 0.85(d, 6H)

[0135] LC / MS [M+H]+ 310.12

[0136] Example 3: Synthesis of 4-benzyl-N-(cyclohexylmethyl)furo[3,2-d]pyrimidine-2-carboxamide

[0137] 1H-NMR (400MHz, DMSO-d6) δ 8.98(s, 1H), 8.13(s, 1H), 7.28-7.20(m, 5H), 6.30(d, 1H), 3.81(s, 2H), 2.88(m, 2H), 2.06(d, 1H), 1.63-1.53(m, 8H), 1.46(d, 2H)

[0138] LC / MS [M+H]+ 350.14

[0139] Example 4: Synthesis of 4-benzyl-N-(morpholinomethyl)furo[3,2-d]pyrimidine-2-carboxamide

[0140] 1H-NMR (400MHz, DMSO-d6) δ 9.38(s, 1H), 8.13(d, 1H), 7.28-7.20(m, 5H), 6.30(d, 1H), 4.03(s, 2H), 3.81(s, 2H), 3.49(t, 4H), 2.40(t, 4H)

[0141] LC / MS [M+H]+ 353.08

[0142] Example 5: Synthesis of 4-benzyl-N-(pyridin-2-ylmethyl)thieno[3,2-d]pyrimidine-2-carboxamide

[0143] 1H-NMR (400MHz, DMSO-d6) δ 9.26(s, 1H), 8.51(d, 1H), 7.74(t, 1H), 7.35(d, 1H), 7.31(d, 1H), 7.28-7.20(m, 6H), 7.12(d, 1H), 4.37(s, 2H), 3.81(s, 2H)

[0144] LC / MS [M+H]+ 361.04

[0145] Example 6: Synthesis of N,4-dibenzylthieno[3,2-d]pyrimidine-2-carboxamide

[0146] 1H-NMR (400MHz, DMSO-d6) δ 9.26(s, 1H), 7.31-7.20(m, 11H), 7.12(d, 1H), 4.11(s, 2H), 3.81(s, 2H)

[0147] LC / MS [M+H]+ 360.04

[0148] Example 7: Synthesis of 4-benzyl-N-(2-chlorobenzyl)thieno[3,2-d]pyrimidine-2-carboxamide

[0149] 1H-NMR (400MHz, DMSO-d6) δ 9.26(s, 1H), 7.68(d, 1H), 7.31(d, 1H), 7.28-7.20(m, 8H), 7.12(d, 1H), 4.23(s, 2H), 3.81(s, 2H)

[0150] LC / MS [M+H]+ 394.01

[0151] Example 8: Synthesis of 4-benzyl-N-((5-methylfuran-2-yl)methyl)thieno[3,2-d]pyrimidine-2-carboxamide

[0152] 1H-NMR (400MHz, DMSO-d6) δ 9.26(s, 1H), 7.31-7.20(m, 6H), 7.12(d, 1H), 6.17(d, 1H), 6.02(d, 1H), 4.46(s, 2H), 3.81(s, 2H), 2.20(d, 3H)

[0153] LC / MS [M+H]+ 364.05

[0154] Example 9: Synthesis of 4-benzyl-N-propylbenzofuro[3,2-d]pyrimidine-2-carboxamide

[0155] 1H-NMR (400MHz, DMSO-d6) δ 8.93(s, 1H), 7.70(d, 1H), 7.65(d, 1H), 7.36-7.20(m, 7H), 3.81(s, 2H), 3.25(t, 2H), 1.55(m, 2H), 0.88(t, 3H)

[0156] LC / MS [M+H]+ 346.11

[0157] Example 10: Synthesis of 4-benzyl-N-isobutylbenzofuro[3,2-d]pyrimidine-2-carboxamide

[0158] 1H-NMR (400MHz, DMSO-d6) δ 8.98(s, 1H), 7.70(d, 1H), 7.65(d, 1H), 7.36(m, 1H), 7.28-7.20(m, 6H), 3.81(s, 2H), 2.88(d, 2H), 2.34(m, 1H), 0.85(d, 6H)

[0159] LC / MS [M+H]+ 360.07

[0160] Example 11: Synthesis of N,4-dibenzylbenzofuro[3,2-d]pyrimidine-2-carboxamide

[0161] 1H-NMR (400MHz, DMSO-d6) δ 9.26(s, 1H), 7.70(d, 1H), 7.65(d, 1H), 7.36-7.20(m, 12H), 4.11(s, 2H), 3.81(s, 2H)

[0162] LC / MS [M+H]+ 394.08

[0163] Example 12: Synthesis of 4-benzyl-N-(2-chlorobenzyl)benzofuro[3,2-d]pyrimidine-2-carboxamide

[0164] 1H-NMR (400MHz, DMSO-d6) δ 9.26(s, 1H), 7.70-7.65(m, 3H), 7.36-7.20(m, 10H), 4.23(s, 2H), 3.81(s, 2H)

[0165] LC / MS [M+H]+ 428.07

[0166] Example 13: Synthesis of 4-benzyl-N-(cyclohexylmethyl)benzo[4,5]thieno[3,2-d]pyrimidine-2-carboxamide

[0167] 1H-NMR (400MHz, DMSO-d6) δ 8.98(s, 1H), 8.05(d, 1H), 7.93(d, 1H), 7.49-7.20(m, 7H), 3.81(s, 2H), 2.88(d, 2H), 2.06(m, 1H), 1.63-1.46(m, 10H)

[0168] LC / MS [M+H]+ 416.14

[0169] Example 14: Synthesis of 4-benzyl-N-(morpholinomethyl)benzo[4,5]thieno[3,2-d]pyrimidine-2-carboxamide

[0170] 1H-NMR (400MHz, DMSO-d6) δ 9.38(s, 1H), 8.05(d, 1H), 7.93(d, 1H), 7.49-7.42(m, 2H), 7.28-7.20(m, 5H), 4.03(s, 2H), 3.81(s, 2H), 3.49(t, 4H), 2.40(t, 4H)

[0171] LC / MS [M+H]+ 419.11

[0172] Example 15: Synthesis of 4-benzyl-N-(pyridin-2-ylmethyl)benzo[4,5]thieno[3,2-d]pyrimidine-2-carboxamide

[0173] 1H-NMR (400MHz, DMSO-d6) δ 9.26(s, 1H), 8.51(d, 1H), 8.05(d, 1H), 7.93(d, 1H), 7.74(m, 1H), 7.49-7.20(m, 9H), 4.37(s, 2H), 3.81(s, 2H)

[0174] LC / MS [M+H]+ 411.07

[0175] Example 16: Synthesis of 4-benzyl-N-((5-methylfuran-2-yl)methyl)benzo[4,5]thieno[3,2-d]pyrimidine-2-carboxamide

[0176] 1H-NMR (400MHz, DMSO-d6) δ 9.26(s, 1H), 8.05(d, 1H), 7.93(d, 1H), 7.49(t, 1H), 7.42(t, 1H), 7.28-7.20(m, 5H), 6.17(d, 1H), 6.02(d, 1H), 4.46(s, 2H), 3.81(s, 2H), 2.20(s, 3H)

[0177] LC / MS [M+H]+ 414.07

[0178] Example 17: Synthesis of 4-benzyl-N-propyl-6,7,8,9-tetrahydrobenzofuro[3,2-d]pyrimidine-2-carboxamide

[0179] 1H-NMR (400MHz, DMSO-d6) δ 8.93(s, 1H), 7.28-7.20(m, 5H), 3.81(s, 2H), 3.25(t, 2H), 2.81(t, 2H), 2.73(t, 2H), 1.87(m, 4H), 1.55(m, 2H), 0.88(t, 3H)

[0180] LC / MS [M+H]+ 350.16

[0181] Example 18: Synthesis of 4-benzyl-N-isobutyl-6,7,8,9-tetrahydrobenzofuro[3,2-d]pyrimidine-2-carboxamide

[0182] 1H-NMR (400MHz, DMSO-d6) δ 8.98(s, 1H), 7.28-7.20(m, 5H), 3.81(s, 2H), 2.88(d, 2H), 2.81(m, 2H), 2.73(m, 2H), 2.34(m, 1H), 1.87(m, 4H), 0.85(d, 6H)

[0183] LC / MS [M+H]+ 364.17

[0184] Example 19: Synthesis of N,4-dibenzyl-6,7,8,9-tetrahydrobenzofuro[3,2-d]pyrimidine-2-carboxamide

[0185] 1H-NMR (400MHz, DMSO-d6) δ 9.26(s, 1H), 7.31-7.20(m, 10H), 4.11(s, 2H), 3.81(s, 2H), 2.81(t, 2H), 2.73(t, 2H), 1.87(m, 4H)

[0186] LC / MS [M+H]+ 398.10

[0187] Example 20: Synthesis of 4-benzyl-N-(2-chlorobenzyl)-6,7,8,9-tetrahydrobenzofuro[3,2-d]pyrimidine-2-carboxamide

[0188] 1H-NMR (400MHz, DMSO-d6) δ 9.26(s, 1H), 7.68(d, 1H), 7.28-7.20(m, 8H), 4.23(s, 2H), 3.81(s, 2H), 2.81(t, 2H), 2.73(t, 2H), 1.87(m, 4H)

[0189] LC / MS [M+H]+ 432.09

[0190] Example 21: Synthesis of 4-benzyl-N-(cyclohexylmethyl)-6,7,8,9-tetrahydrobenzo[4,5]thieno[3,2-d]pyrimidine-2-carboxamide

[0191] 1H-NMR (400MHz, DMSO-d6) δ 8.98(s, 1H), 7.28-7.20(m, 5H), 3.81(s, 2H), 2.88(d, 2H), 2.78(t, 2H), 2.73(t, 2H), 2.06(m, 1H), 1.78(m, 4H), 1.63-1.46(m, 10H)

[0192] LC / MS [M+H]+ 420.11

[0193] Example 22: Synthesis of 4-benzyl-N-(morpholinomethyl)-6,7,8,9-tetrahydrobenzo[4,5]thieno[3,2-d]pyrimidine-2-carboxamide

[0194] 1H-NMR (400MHz, DMSO-d6) δ 9.38(s, 1H), 7.28-7.20(m, 5H), 4.03(s, 2H), 3.81(s, 2H), 3.49(t, 4H), 2.78(t, 2H), 2.73(t, 2H), 2.40(t, 4H), 1.78(m, 4H)

[0195] LC / MS [M+H]+ 423.10

[0196] Example 23: Synthesis of 4-benzyl-N-(pyridin-2-ylmethyl)-6,7,8,9-tetrahydrobenzo[4,5]thieno[3,2-d]pyrimidine-2-carboxamide

[0197] 1H-NMR (400MHz, DMSO-d6) δ 9.26(s, 1H), 8.51(d, 1H), 7.74(t, 1H), 7.35(d, 1H), 7.28-7.20(m, 6H), 4.37(s, 2H), 3.81(s, 2H), 2.78(m, 2H), 2.73(m, 2H), 1.78(m, 4H)

[0198] LC / MS [M+H]+ 415.08

[0199] Example 24: Synthesis of 4-benzyl-N-((5-methylfuran-2-yl)methyl)-6,7,8,9-tetrahydrobenzo[4,5]thieno[3,2-d]pyrimidine-2-carboxamide

[0200] 1H-NMR (400MHz, DMSO-d6) δ 9.26(s, 1H), 7.28-7.20(m, 5H), 6.17(d, 1H), 6.02(d, 1H), 4.46(s, 2H), 3.81(s, 2H), 2.78(m, 2H), 2.73(m, 2H), 2.20(s, 3H), 1.78(m, 4H)

[0201] LC / MS [M+H]+ 418.10

[0202] Example 25: Synthesis of 4-benzyl-N-propylpyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0203] 1H-NMR (400MHz, DMSO-d6) δ 8.93(s, 1H), 8.51(d, 1H), 8.43(d, 1H), 7.36(t, 1H), 7.28-7.20(m, 5H), 3.81(s, 2H), 3.25(t, 2H), 1.55(m, 2H), 0.88(t, 3H)

[0204] LC / MS [M+H]+ 347.05

[0205] Example 26: Synthesis of 4-benzyl-N-isobutylpyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0206] 1H-NMR (400MHz, DMSO-d6) δ 8.98(s, 1H), 8.51(d, 1H), 8.43(d, 1H), 7.36(t, 1H), 7.28-7.20(m, 5H), 3.81(s, 2H), 2.88(d, 2H), 2.34(m, 1H), 0.85(d, 6H)

[0207] LC / MS [M+H]+ 361.08

[0208] Example 27: Synthesis of N,4-dibenzylpyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0209] 1H-NMR (400MHz, DMSO-d6) δ 9.26(s, 1H), 8.51(d, 1H), 8.43(d, 1H), 7.36-7.20(m, 11H), 4.11(s, 2H), 3.81(s, 2H)

[0210] LC / MS [M+H]+ 395.11

[0211] Example 28: Synthesis of 4-benzyl-N-(morpholinomethyl)pyrido[3',2':4,5]furo[3,2-d]pyrimidine-2-carboxamide

[0212] 1H-NMR (400MHz, DMSO-d6) 9.26(s, 1H), 8.51(d, 1H), 8.43(d, 1H), 7.68(d, 1H), 7.36-7.20(m, 9H), 4.23(s, 2H), 3.81(s, 2H)

[0213] LC / MS [M+H]+ 429.07

[0214] Example 29: Synthesis of 4-benzyl-N-(cyclohexylmethyl)pyrido[3',2':4,5]thieno[3,2-d]pyrimidine-2-carboxamide

[0215] 1H-NMR (400MHz, DMSO-d6) δ 8.98(s, 1H), 8.67(d, 1H), 8.51(d, 1H), 7.36(t, 1H), 7.28-7.20(m, 5H), 3.81(s, 2H), 2.88(d, 2H), 2.06(m, 1H), 1.63-1.46(m, 10H)

[0216] LC / MS [M+H]+ 417.12

[0217] Example 30: Synthesis of 4-benzyl-N-(morpholinomethyl)pyrido[3',2':4,5]thieno[3,2-d]pyrimidine-2-carboxamide

[0218] 1H-NMR (400MHz, DMSO-d6) δ 9.38(s, 1H), 8.67(d, 1H), 8.51(d, 1H), 7.36(t, 1H), 7.28-7.20(m, 5H), 4.03(s, 2H), 3.81(s, 2H), 3.49(t, 4H), 2.40(t, 4H)

[0219] LC / MS [M+H]+ 420.07

[0220] Example 31: Synthesis of 4-benzyl-N-(pyridin-2-ylmethyl)pyrido[3',2':4,5]thieno[3,2-d]pyrimidine-2-carboxamide

[0221] 1H-NMR (400MHz, DMSO-d6) δ 9.26(s, 1H), 8.67(d, 1H), 8.51(m, 1H), 8.51(m, 1H), 7.74(t, 1H), 7.36(m, 2H), 7.28-7.20(m, 6H), 4.37(s, 2H), 3.81(s, 2H)

[0222] LC / MS [M+H]+ 412.05

[0223] Example 32: Synthesis of 4-benzyl-N-((5-methylfuran-2-yl)methyl)pyrido[3',2':4,5]thieno[3,2-d]pyrimidine-2-carboxamide

[0224] 1H-NMR (400MHz, DMSO-d6) δ 9.26(s, 1H), 8.67(d, 1H), 8.51(d, 1H), 7.36(t, 1H), 7.28-7.20(m, 5H), 6.17(d, 1H), 6.02(d, 1H), 4.46(s, 2H), 3.81(s, 2H), 2.20(s, 3H)

[0225] LC / MS [M+H]+ 415.05

[0226]

[0227] [Example Exam]

[0228] Test Example 1: Binding Test of KRAS G12D Protein and Compound

[0229] To confirm the binding affinity of the example compound to the KRAS G12D protein, the following experiment was conducted using an OCTET device.

[0230] The OCTET instrument is a biomolecular reaction analyzer that utilizes Bio-Layer Interferometry (BLI) technology to measure intermolecular interactions in real time without the need for separate fluorescent labels. Using this technology, the binding and dissociation phenomena of proteins and compounds were measured, thereby identifying binding affinities between molecules.

[0231] KRAS G12D protein was purchased and used as a recombinant protein produced by Genomine Co., Ltd. KRAS G12D recombinant protein was biotinylated and used for OCTET binding assay. 200 μL of Buffer (PBS + 1% DMSO + 0.02% Tween 20), Ligand (1 / 10 KRAS G12D + Buffer), Quench (EZ-Link + Buffer), and compounds at different concentrations were dispensed per well on a Grainer black plate. The binding was measured in the order of Buffer-Loading (Ligand or Buffer) - Quenching - Buffer - Buffer (Baseline) - Compound - Buffer using the SSA sensor of the OCTET device. K, which removed nonspecific binding through the OCTET program, D The value was derived.

[0232] MRTX1133 (CAS No.: 2621928-55-8) of Comparative Example 1 was purchased and used from MedChemExpress.

[0233] [Table 3]

[0234]

[0235]

[0236] As shown in the above [Table 3], the compound according to the embodiment of the present invention specifically binds to the KRAS G12D protein, and it was confirmed that the binding affinity to the KRAS G12D protein was higher than that of the material of Comparative Example 1.

[0237] Test Example 2: In vitro anticancer efficacy test on pancreatic cancer cell lines

[0238] IC using pancreatic cancer cell lines ASPC-1 and HPAF-ll with KRAS G12D mutation 50The anticancer effect of the example compound was confirmed by CCK assay. ASPC-1 cell line was purchased from Korea Cell Line Bank (KCLB), and cell culture was performed using RPMI1640 media. Cell suspension was prepared, AsPC-1 cell line was seeded at 3000c / w, and cultured in a CO2 incubator for 24 hours. Each substance was dissolved in DMSO at a high concentration to prepare a stock solution, and then diluted to a maximum of 0.1% in the medium to prepare. After completion of culture, the prepared substance was treated at each concentration, and the absorbance was examined after 3 days to determine the IC of the sample. 50 The values ​​were derived. The number of n per sample was 3 or more. The HPAF-II cell line was purchased from the Korea Cell Line Bank (KCLB), and the cells were cultured using MEM. After making a cell suspension, the HPAF-II cell line was seeded at 3000 c / w and cultured in a CO2 incubator for 24 hours. Each substance was dissolved in DMSO at a high concentration to prepare a stock solution, and then diluted to a maximum of 0.1% in the medium to prepare. After the culture was completed, the prepared substances were treated at each concentration, and the absorbance was examined after 3 days to determine the IC of the sample. 50 The value was derived. The number of n per sample was 3 or more.

[0239] [Table 4]

[0240]

[0241]

[0242] As shown in the above [Table 4], the example compound of the present invention has an IC similar to or lower than that of Comparative Example 1 in the KRAS G12D mutant pancreatic cancer cell line. 50 It was confirmed that the compound of the present invention has excellent anticancer activity by specifically binding to the KRAS G12D protein and inhibiting cell growth in a related pancreatic cancer cell line.

Claims

1. A compound of the following formula I, a solvate, a stereoisomer or a pharmaceutically acceptable salt thereof: [Chemical Formula I] In the above formula, X is O or S, R1 and R2 are each independently H, or R1 and R2 are combined with each other to form a 6-membered ring, R3 is C 1-6 Alkyl, Phenyl, C 3-6 Cycloalkyl, C 3-6 Heterocycloalkyl or C 3-6 It is heteroaryl, C above 1-6 Alkyl, Phenyl, C 3-6 Cycloalkyl, C 3-6 Heterocycloalkyl or C 3-6 At least one hydrogen of the heteroaryl is replaced with one or more substituents selected from the group consisting of halogen and carbon.

2. In claim 1, A compound of formula I, a solvate, a stereoisomer or a pharmaceutically acceptable salt thereof, wherein the 6-membered ring formed by combining R1 and R2 is selected from the group consisting of the following structural formulae 1 to 8: .

3. In claim 1, C above 1-6 Alkyl is any one selected from the group consisting of ethyl and isopropyl, C above 3-6 Cycloalkyl is cyclohexyl, C above 3-6 Heterocycloalkyl is morpholinyl, C above 3-6 A compound of formula I, a solvate, a stereoisomer or a pharmaceutically acceptable salt thereof, wherein heteroaryl is any one selected from the group consisting of pyridinyl and furanyl.

4. In claim 1, A compound of formula I, a solvate, a stereoisomer or a pharmaceutically acceptable salt thereof, selected from the group consisting of compounds represented by the following chemical formulas: .

5. In claim 1, A compound of formula I, a solvate, a stereoisomer or a pharmaceutically acceptable salt thereof, selected from the group consisting of compounds represented by the following chemical formulas: .

6. A pharmaceutical composition for preventing or treating cancer, comprising a compound, a solvate, a stereoisomer or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5 as an active ingredient.

7. In claim 6, A pharmaceutical composition for preventing or treating cancer, wherein the cancer comprises tumor tissue in which a KRAS G12D mutation exists.

8. In claim 6, A pharmaceutical composition for preventing or treating 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, colon cancer, colon cancer, skin cancer, head and neck cancer, melanoma, and thyroid cancer.

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