Novel selenophene derivatives

Abstract

The present invention provides compounds defined by chemical formula 1 or tautomers thereof, and pharmaceutically acceptable salts thereof. The compounds of the present invention can inhibit the activity of SOS1.

Description

Novel selenophene derivatives

[0001] The present invention relates to small organic compounds having inhibitory activity against biological enzymes. The present invention also relates to pharmaceutical compositions containing such small organic compounds.

[0002] SOS1 (Son of Sevenless 1) is known to play a key role in activating small GTPases, such as Rat Sarcoma viral oncogene homolog (RAS) and Ras-related C3 botulinum toxin substrate 1 (Rac1). These small GTPases are important components of cell signaling pathways that regulate various cellular processes, such as cell growth, differentiation, and migration.

[0003] SOS1 acts as an active guanine nucleotide exchange factor (GEF) that catalyzes the exchange of GDP (guanosine diphosphate)-bound Ras to GTP (guanosine triphosphate)-bound Ras, thereby activating RAS. GTP-bound activated RAS then interacts with downstream enzyme proteins to initiate signaling pathways. RAS activation by SOS1 is a crucial step in various cellular processes, including cell proliferation, survival, and differentiation. Activated RAS initiates downstream signaling pathways, such as the mitogen-activated protein kinase (MAPK) pathway, which regulates gene expression and the cell's response to external cues. Deregulation of the RAS pathway is associated with mutations in tumors, making it a key target in cancer research. There are three isoforms of RAS, KRAS, NRAS, and HRAS, of which KRAS is the most frequently mutated, and SOS1 is known to be crucial for the activation of mutant KRAS and oncogenic signaling (Jeng et al., Nat. Commun., 2012, 3:1 168). In particular, depletion of intracellular SOS1 concentrations decreased the proliferation rate and viability of tumor cells harboring KRAS mutations, but had no effect on cell lines harboring the wild-type KRAS gene. In addition, the damage caused by SOS1 deletion was observed to be irreversible by introducing SOS1 with a mutated catalytic active site, demonstrating that the GEF activity of SOS1 plays a key role in KRAS mutant cancer cells.

[0004] In addition to RAS, SOS1 participates in the activation of Rac1. Rac1, another small GTPase belonging to the Rho protein family, plays a crucial role in regulating the cellular cytoskeleton, migration, and other processes. SOS1 converts GDP bound to Rac1 into GTP, thereby converting it into an active state. Activated Rac1 is known to play a crucial role in processes such as cell adhesion, migration, and invasion, playing a crucial role in normal cell function and in pathological conditions such as cancer metastasis.

[0005] In view of these points, SOS1 inhibitors may be developed as therapeutic drugs for diseases caused by KRAS mutations or Rac1 activation.

[0006] One object of the present invention is to provide a novel selenophene derivative compound that is an SOS1 inhibitor with excellent activity in inhibiting KRAS-SOS1 protein-protein interaction. Another object of the present invention is to provide a pharmaceutical composition comprising the compound in a therapeutically effective amount.

[0007] In one aspect of the present invention, a selenophene compound defined by chemical formula 1 having an inhibitory activity on SOS1 or a totoisomer thereof and a pharmaceutically acceptable salt thereof are provided.

[0008] [Chemical Formula 1]

[0009]

[0010] In the above chemical formula 1, R 1 and R 2 are H and C respectively 1~4 Alkyl and deuterium substituted C 1~4 A functional group independently selected from the group consisting of alkyl, or R 1 and R 2 is R 1 and R 2 Together with the nitrogen atom to which it is bonded, it forms a 4-6 membered heterocycloalkyl ring, where R1 and R 2 Except when both are H at the same time,

[0011] Is

[0012]

[0013] And at this time Q 1 and Q 2 are each independently C 1~4 A functional group selected from alkyl and 4-8 membered single ring heterocyclyl, and Q 3 is a 4-8 membered single ring heterocyclyl, The sign is It refers to the connection at that position to the rest of the compound.

[0014] In another aspect of the present invention, there is provided a pharmaceutical composition for the treatment of cancer comprising a therapeutically effective amount of the compound described above, a totoisomer thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0015] In another aspect of the present invention, a method for treating cancer is provided. The method comprises administering to a patient a therapeutically effective amount of a compound of the present invention, such as a compound defined by Formula 1, or a pharmaceutical composition comprising such a compound of the present invention.

[0016] The compound of the present invention can inhibit the activity of SOS1. Furthermore, the compound of the present invention exhibits a synergistic cell death effect in conjunction with blockade of RAS signaling or EGFR signaling. In a preferred embodiment, the compound of the present invention exhibits selective inhibitory activity against SOS1 over SOS2. The compound of the present invention is useful for the treatment of cancer.

[0017] Figure 1 is a dose-response curve for cell growth inhibition showing the synergistic effect observed when the compound of the present invention is used in combination with a targeted anticancer agent that blocks RAS signaling in the NCI-H358 cancer cell line.

[0018] Figure 2 is a dose-response curve for cell growth inhibition showing the synergistic effect that occurs when the compound of the present invention is used in combination with a targeted anticancer agent that blocks EGFR signaling in the PC-9 cancer cell line.

[0019] Figure 3 is a graph showing that the compound of the present invention increases the production of reactive oxygen species in MIA-PaCa-2 cancer cells.

[0020] Hereinafter, 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 concept of a term to best explain his or her invention, they should be interpreted in a way that aligns with the technical spirit of the present invention.

[0021] Therefore, the embodiments described in this specification are merely examples presented for the purpose of helping to understand the present invention and do not represent all of the technical ideas of the present invention. Therefore, it should be understood that there may be various equivalents and modified examples that can replace them at the time of filing this application.

[0022] Unless otherwise defined, all technical terms used herein have the same meaning as commonly understood by those skilled in the art. While preferred methods and samples are described herein, similar or equivalent methods are also included within the scope of the present invention. Numerical values ​​described herein are considered to include the meaning of "about" even if not explicitly stated. The numerical range indicated using the term "to" herein includes the ranges that include the values ​​described before and after the term "to" as the lower and upper limits, respectively.

[0023] Definition of terms

[0024] In this specification, the term "inhibits SOS1" or "inhibits SOS1" or "inhibits SOS1 activity" or "inhibits SOS1 activity" or the term "SOS1 inhibitor" or "SOS1 inhibitor" refers to the action of the compound in reducing the guanine nucleotide exchange activity of SOS1 for RAS or blocking the binding of RAS to SOS1. For example, an "SOS1 inhibitor" or "SOS1 inhibitor" is a substance that has the effect of reducing the measured signal of guanine nucleotide exchange activity of SOS1 or the measured signal of binding of RAS and SOS1 by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 95% when measured under comparable conditions with a negative control (e.g., a blank).

[0025] Those skilled in the art will appreciate that some of the compounds of the present invention may exist in more than one isomeric form. Since a chemical structural formula can only represent one isomeric form, it is well understood that when a compound is referred to by a single structural formula for convenience, the structural formula also encompasses the isomeric variations of that compound. Depending on the compound, one of several isomeric forms may exist primarily, a mixture of multiple isomeric forms may exist at room temperature, or only one isomeric form may be isolated. Examples of isomeric forms include those between the pyridone and hydroxypyridine forms, and those between the keto and enol forms.

[0026] In this specification, the prefix "C x~y " or "C x ~C y"(where x and y are natural numbers) is used in front of a functional group to indicate the number of carbon atoms that the functional group has in its backbone. For example, C 1~4 An alkyl group has 1 to 4 carbon atoms, C 1~3 An alkyl group refers to an alkyl group having 1 to 3 carbon atoms. For example, C1~C4 alkoxy groups include methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, sec-butoxy, and t-butoxy. For example, C3~C6 cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl groups. In addition, in the present specification, when the number of elements forming the ring or the number of rings is indicated in front of a ring functional group or compound (including both a carbon ring and a heterocycle), the ring is a ring formed of the corresponding number of elements, that is, carbon and / or heteroatoms, or a structure having the corresponding number of rings. For example, a six-membered ring heteroaryl refers to a heteroaryl group that contains one or more heteroatoms and is formed together with carbon and has 6 ring elements. Also, for example, a 4-6 membered single ring heteroaryl is a monovalent aromatic ring composed of only one ring, not a fused ring, bridged ring, or spiro ring compound, and the total number of carbons and heteroatoms forming the ring skeleton is 4 to 6, such as pyrrolyl, pyridinyl, imidazolyl, isoxazolyl, thiophenyl, and thiazolyl.

[0027] Unless otherwise specified herein, when a functional group is linked to another part of a compound by a bond, the linkage may be via any atom of that functional group, provided that it is a suitable atom. For example, when referring to a propyl group, both prop-1-yl and prop-2-yl are included.

[0028] As used herein, the term "halogen" refers to an atom selected from fluorine, chlorine, bromine and iodine.

[0029] In this specification, "C 1~6 The term "alkyl" refers to a group whose chemical formula is C when n is a natural number from 1 to 6. n H 2n+1 It refers to a saturated hydrocarbon group of 1-valent. C 1~6 Alkyl groups include all leading or branched saturated hydrocarbons having 1 to 6 carbon atoms, such as n-propyl, i-propyl, 2-methyl-ethyl, sec-butyl, tert-butyl, etc.

[0030] In this specification, "C 1~6 The term "alkylene" refers to a group whose chemical formula is C when n is a natural number from 1 to 6. n H 2n It refers to a saturated hydrocarbon group of two atoms.

[0031] The term "cycloalkyl" as used herein refers to a monovalent saturated hydrocarbon atom group containing one or more carbon rings. A monocyclic cycloalkyl containing only one carbon ring has a chemical formula of C n H 2n-1 is a single ring C 3~6 Examples of cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0032] As used herein, the term "bicyclic" refers to a carbon ring compound or heterocyclic compound composed of two rings, which is formed by sharing one or more ring atoms between the two constituent rings. In a bicyclic compound, the atom shared between the two rings is called a bridgehead atom. Bicyclic compounds are divided into three types depending on the number of bridgehead atoms and whether or not there is a direct connection between the bridgehead atoms. A fused bicyclic compound has two bridgehead atoms, which are directly connected to each other by a bond. Examples of fused bicyclic compounds include decalin, naphthalene, anthracene, phenanthrene, indole, benzofuran, purine, and quinoline. A ring compound with only one bridgehead atom is a spirocyclic compound. A bridged ring compound has two bridgehead atoms that are not directly connected to each other by a bond, but rather have one or more ring atoms interposed between the two bridgehead atoms. Examples of bridged ring compounds include norbornane, 7-oxabicyclo[2.2.1]heptane, and adamantane.

[0033] In this specification, fused bicyclic compounds can be expressed by specifying the number of ring atoms in each of the two rings that make up the compound. For example, thienopyridine, benzofuran, indole, etc. can be referred to as five- or six-membered fused bicyclic compounds. Similarly, naphthalene, chromane, tetrahydroquinoline, quinoline, quinoxaline, pteridine, etc. can be referred to as six- or six-membered fused bicyclic compounds.

[0034] The term "carbocyclyl" as used herein refers to a hydrocarbon group having a saturated or unsaturated ring that is not aromatic, has a ring size of 3 to 14 members, and a monovalent atomic group whose ring skeleton is composed of only carbon atoms without a heteroatom. As used herein, saturated carbocyclyl includes not only a monocyclic functional group but also a multi-ring saturated carbocyclyl, for example, a bicyclic hydrocarbon group of a fused ring, a bridged ring, and a spiro ring saturated hydrocarbon ring. For example, exemplary saturated C 3~10 Some examples of carbocyclyl groups include the aforementioned cycloheptyl (C7), cyclooctyl (C8), bicyclo[1.1.1]pentanyl (C5), bicyclo[2.2.1]heptanyl (C7), bicyclo[2.2.2]octanyl (C8), cyclononyl (C9), cyclodecyl (C 10 ), decahydronaphthalenyl(decalinyl)(C 10 ), Adamantyl (C 10 ), spiro[4.5]decanyl(C 10 ) etc. Unsaturated carbocyclyl refers to a hydrocarbon ring that has one or more double bonds or triple bonds in the ring and is not aromatic. Examples of unsaturated carbocyclyl include cyclopentadienyl, cyclohexenyl, and norbornenyl.

[0035] The term "aryl" as used herein refers to a monovalent hydrocarbon functional group having an aromatic hydrocarbon ring of 6 to 14 ring carbon atoms and optionally containing one to three fused rings. Examples of aryl include phenyl, naphthyl, anthracenyl, and phenanthrenyl.

[0036] The term "heteroaryl" as used herein refers to an aromatic ring which is a monovalent 5 to 12 membered unsaturated ring, which comprises carbon atoms and one or more heteroatoms selected from nitrogen, oxygen and sulfur in the ring skeleton. The ring system of the heteroaryl may be a single ring, or a dicyclic or tricyclic ring in the form of a fused ring. The term heteroaryl excludes rings which contain a direct link between oxygen and sulfur atoms, such as ―O―O―, ―O―S― or ―S―S―, among the ring constituent atoms. Some examples of heteroaryl include pyrrolyl, imidazolyl, pyrazolyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, pyrimidyl, triazolyl, quinazolinyl, 2-furanyl, 3-furanyl, benzofuranyl, 2-thienyl, oxazolyl, isothiazolyl, and thiadiazolyl. Heteroaryl can also be defined by specifying its heteroatom; for example, a heteroaryl in which the heteroatom is nitrogen would include pyrrolyl, imidazolyl, pyrazolyl, and 2-pyridyl, but would exclude furanyl and thienyl.

[0037] Fused bicyclic heteroaryl groups include six- and five-membered fused bicyclic heteroaryl groups such as benzimidazolyl, benzofuranyl, benzothiophenyl, isoindolyl, indazolyl, imidazopyridinyl, imidazopyrimidinyl, imidazopyridazinyl, indazolyl, pyrazolopyridinyl, pyrrolopyrimidinyl, pyrrolopyridinyl, pyrrolopyrazinyl, triazolopyridinyl, triazolopyrimidinyl, purinyl, benzothiazolyl, benzoisothiazolyl, benzothiadiazolyl, benzoxazolyl, benzothiadiazolyl, etc., and six- and six-membered fused bicyclic heteroaryl compounds such as benzopyridinyl, benzopyrimidinyl, quinolinyl, isoquinolinyl, phthalazinyl, Examples include quinazolinyl, quinoxalinyl, naphthyridinyl, quinolinzinyl, and cinnolinyl.

[0038] The term "heteroarylene" as used herein refers to a divalent aromatic ring formed by removing one hydrogen atom from the aforementioned heteroaryl.

[0039] The term "heterocycloalkyl" as used herein refers to a saturated ring that is non-aromatic and is a monovalent atomic group in which at least one of the carbon atoms forming the ring in the cycloalkyl is independently substituted with a heteroatom selected from nitrogen, oxygen, and sulfur. Examples of heterocycloalkyl include azetidinyl, pyrrolidinyl, imidazolidinyl, piperidinyl, tetrahydrofuranyl, piperazinyl, morpholinyl, and thiazolidinyl.

[0040] The term "monocyclic heterocyclyl" as used herein refers to a monovalent unsaturated or saturated monocyclic ring, which includes at least one heteroatom selected from nitrogen, oxygen and sulfur among the ring skeletal atoms. Therefore, monocyclic heterocyclyl also includes monocyclic heteroaryl, monocyclic heterocycloalkyl and monocyclic heterocycles that include unsaturated bonds and heteroatoms in the ring skeleton but are not aromatic.

[0041] In this specification, the term "substituted" or "substituted" in front of a functional group refers to a structure in which at least one of the hydrogens of the functional group is replaced with a functional group other than the designated hydrogen, unless otherwise stated in the specification, provided that the substituted functional group maintains its normal valence and yields a chemically stable functional group. In typical cases, the substituent is a halogen, CN, OH, C 1~6 Alkyl, C 3~6 Cycloalkyl, C 3~14 Heteroaryl, C 1~6 Choose between alkoxy and CF3.

[0042] In this specification, a specific atom of a substituent forming part of a compound is " " indicates that the substituent is chemically bonded to the rest of the compound through that atom.

[0043] In this specification, if a dotted line such as "---" is shown on a bond line, it indicates that the bond line is an optional element that may or may not be present. For example, A notation like this indicates that this structural formula encompasses benzene and 1,3-cyclohexadiene.

[0044] As used herein, the term "pharmaceutically acceptable salt" for a compound refers to a salt that does not impair the desired biological activity of the compound and does not cause inappropriate toxicity, irritation, or allergic reactions when used in contact with human or animal tissues. Pharmaceutically acceptable salts are well known in the art. For example, reference may be made to prior literature, such as the paper by Berge et al., J. Pharmaceutical Sciences (1977), Vol. 66, pp. 1-19. Some examples of pharmaceutically acceptable salts include acid addition salts such as hydrochloride, hydrobromide, phosphate, sulfate, perchlorate, acetate, oxalate, maleate, tartrate, citrate, succinate, malonate, adipate, alginate, ascorbate, aspartate, benzoate, benzenesulfonate, and hydrosulfide, and base addition salts such as alkali metal salts, alkaline earth metal salts, ammonium salts, and quaternary ammonium (N + (C 1~4 There are alkyl)4), and examples of alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, and magnesium.

[0045] compound

[0046] In one aspect of the present invention, a compound defined by chemical formula 1, its isomers, and pharmaceutically acceptable salts thereof are provided.

[0047]

[0048] In the above chemical formula 1, R 1 and R 2 are H and C respectively 1~4 Alkyl and deuterium substituted C 1~4 A functional group independently selected from the group consisting of alkyl, or R 1 and R 2 is R 1 and R 2 Together with the nitrogen atom to which it is bonded, it forms a 4-6 membered heterocycloalkyl ring, where R 1 and R 2 Except when both are H at the same time,

[0049] Is

[0050]

[0051] And at this time Q 1 and Q 2 are each independently C 1~4 A functional group selected from alkyl and 4-8 membered single ring heterocyclyl, and Q 3 is a 4-8 membered single ring heterocyclyl, The sign is It refers to the connection at that position to the rest of the compound.

[0052] In one specific embodiment of the present invention, the compound of formula 1, R described above 1 and R 2 One of them is hydrogen, and the other is a methyl group or trideuteriomethyl (CD3) group.

[0053] In another embodiment of the present invention, the 4-6 membered heterocycloalkyl ring in the compound of formula 1 is azetidinyl or pyrrolidinyl.

[0054] In another embodiment of the present invention, Q in the compound of formula 1 1 and Q 2 are all methyl, or Q 1and Q 2 One of them is methyl and the other is tetrahydrofuranyl.

[0055] In another embodiment of the present invention, Q in the compound of formula 1 3 All of these are morpholinyl. In a more specific embodiment, the morpholinyl is N-morpholinyl.

[0056] In a more specific embodiment, the compounds of the present invention are the following compounds, stereoisomers, isomers, solvates or pharmaceutically acceptable salts thereof:

[0057] (R)-6,7-dimethoxy-2-methyl-N-(1-(4-(2-((methylamino)methyl)phenyl)selenophen-2-yl)ethyl)quinazolin-4-amine;

[0058] 7-Methoxy-2-methyl-N-((R)-1-(4-(2-((methylamino)methyl)phenyl)selenophen-2-yl)ethyl)-6-(((S)-tetrahydrofuran-3-yl)oxy)quinazolin-4-amine;

[0059] (R)-(2-chloro-4-((1-(4-(2-((methylamino)methyl)phenyl)selenophene-2-yl)ethyl)amino)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(4-methoxytetrahydro-2H-pyran-4-yl)methanone;

[0060] (R)-4-methyl-N-(1-(4-(2-((methylamino)methyl)phenyl)selenophen-2-yl)ethyl)-7-morpholinopyrido[3,4-d]pyridazin-1-amine;

[0061] (R)-N-(1-(4-(2-((dimethylamino)methyl)phenyl)selenophene-2-yl)ethyl)-6,7-dimethoxy-2-methylquinazolin-4-amine;

[0062] (R)-6,7-dimethoxy-2-methyl-N-(1-(4-(2-(pyrrolidin-1-ylmethyl)phenyl)selenophen-2-yl)ethyl)quinazolin-4-amine;

[0063] (R)-6,7-dimethoxy-2-methyl-N-(1-(4-(2-(((methyl-d3)amino)methyl)phenyl)selenophen-2-yl)ethyl)quinazolin-4-amine;

[0064] N-((R)-1-(4-(2-((dimethylamino)methyl)phenyl)selenophene-2-yl)ethyl)-7-methoxy-2-methyl-6-(((S)-tetrahydrofuran-3-yl)oxy)quinazolin-4-amine or

[0065] 7-Methoxy-2-methyl-N-((R)-1-(4-(2-(pyrrolidin-1-ylmethyl)phenyl)selenophen-2-yl)ethyl)-6-(((S)-tetrahydrofuran-3-yl)oxy)quinazolin-4-amine.

[0066] The compound of the present invention can inhibit the activity of SOS1. More specifically, the compound of the present invention can block the interaction between SOS1 and RAS, thereby blocking RAS activation.

[0067] Synthesis of the compound of the present invention

[0068] Compounds of the present invention, including compounds of formula 1, can be synthesized by synthetic routes including those well known in the chemical arts, particularly in light of the teachings contained herein. Starting materials are generally available from commercial sources, such as Aldrich Chemicals (Milwaukee, WI), or are readily prepared using methods well known to those skilled in the art (see, for example, methods generally described in Louis F. Fieser and Mary Fieser, Reagents for Organic Synthesis, 1st to 19th eds., Wiley, NY (1967-1999) or Beilsteins Handbuch der organischen Chemie, 4, Aufl., Springer-Verlag, Berlin, Germany).

[0069] For illustrative purposes, general methods for preparing the compounds and key intermediates of the present invention are outlined in the reaction schemes below. For more detailed descriptions of individual reaction steps, see the specific descriptions in the Examples below. Those skilled in the art will recognize that other synthetic routes can also be used to synthesize the compounds of the present invention. While specific starting materials and reagents are depicted in the reaction schemes and discussed below, other starting materials and reagents can be readily substituted to provide a variety of derivatives and / or reaction conditions. Furthermore, most compounds prepared by the methods described below can be further modified in light of the present disclosure using conventional chemistry well known to those skilled in the art.

[0070] The compounds of the present invention may utilize a deuterium-enriched form as a substituent or functional group. Such deuterated forms can be obtained using the methods described in U.S. Patent Nos. 5,846,514 and 6,334,997, and also by Dean, Dennis C., ed., Curr., Pharm. Des., 2000; 6(10). <Recent Advances in the Synthesis and Applications of Radiolabeled Compounds for Drug Discovery and Development> . Deuterium-enriched starting materials are commercially available and can be applied to the synthetic methods described herein to obtain deuterium-substituted functional groups.

[0071] pharmaceutical composition

[0072] In another aspect, the present invention discloses a pharmaceutical composition comprising the aforementioned compound and a pharmaceutically acceptable excipient. The pharmaceutical composition of the present invention can be used to inhibit the guanine nucleotide exchange activity of SOS1 for RAS in vivo, for example, for the treatment of cancer. In one embodiment, the pharmaceutical composition of the present invention can be used for the treatment of non-small cell lung cancer. In a more specific embodiment, the pharmaceutical composition of the present invention can be used for the treatment of cancer, including KRAS mutant cancer.

[0073] In the pharmaceutical composition of the present invention, the aforementioned compounds may be present as pharmaceutically acceptable salts. Pharmaceutically acceptable salts include, for example, base addition salts and acid addition salts. Pharmaceutically acceptable base addition salts may be formed with metals or amines, such as alkali and alkaline earth metals or organic amines. Pharmaceutically acceptable salts of the compounds may also be prepared using pharmaceutically acceptable cations. Pharmaceutically acceptable acid addition salts include inorganic or organic acid salts. Examples of acid addition salts include those derived from hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, acetic acid, glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, malic acid, mandelic acid, tartaric acid, citric acid, ascorbic acid, palmitic acid, maleic acid, benzoic acid, hydroxybenzoic acid, phenylacetic acid, cinnamic acid, salicylic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, or toluenesulfonic acid.

[0074] In the present invention, a pharmaceutically acceptable excipient refers to an inactive ingredient approved by a relevant administrative agency (e.g., the Ministry of Food and Drug Safety of the Republic of Korea, the Federal Food and Drug Administration (FDA) of the United States) as being suitable for use together with an active pharmaceutical ingredient for the purpose of manufacturing a medicine for treating a disease in humans or livestock. Such pharmaceutically acceptable excipients include, but are not limited to, carriers, lubricants, fluidizing agents, disintegrating agents, sweeteners, diluents, preservatives, coloring agents, flavoring agents, surfactants, wetting agents, dispersing agents, suspending agents, stabilizers, isotonic agents, solvent emulsifiers, and adjuvants.

[0075] The pharmaceutical composition of the present invention may be in a form suitable for oral administration, such as a tablet, capsule, pill, powder, sustained-release preparation, solution, or suspension; in a form suitable for parenteral injection, such as a sterile solution, suspension, or emulsion; in a form suitable for topical administration, such as an ointment or cream; or in a form suitable for rectal administration, such as a suppository.

[0076] The pharmaceutical composition according to the present invention can be prepared in a conventional manner, for example, by conventional mixing, dissolving, granulating, sugar-coated tablet preparation, powdering, emulsifying, encapsulating, encapsulating, or lyophilizing processes. The appropriate formulation will vary depending on the chosen route of administration.

[0077] Pharmaceutical compositions suitable for oral administration can be readily formulated by combining the compounds disclosed herein with pharmaceutically acceptable excipients, such as carriers well known in the art. Using such excipients and carriers, the compounds disclosed herein can be formulated as tablets, pills, dragees, capsules, solutions, gels, syrups, slurries, suspensions, and the like for oral ingestion by patients to be treated. Oral pharmaceutical preparations can be obtained by adding the compounds of the present invention together with solid excipients, grinding the resulting mixture if necessary, adding suitable auxiliaries if necessary, and then processing the granulated mixture to form tablets or dragee cores. Suitable excipients include, for example, fillers and cellulose preparations. If desired, a disintegrant may be added.

[0078] For pharmaceutical compositions for oral administration of a therapeutically effective amount of a compound of the present disclosure, the compositions are generally in the form of solid (e.g., tablets, capsules, pills, powders, or troches) or liquid formulations (e.g., aqueous suspensions, solutions, elixirs, or syrups).

[0079] When administered in tablet form, the composition may additionally contain functional solids and / or solid carriers such as gelatin or adjuvants. Compositions in the form of tablets, capsules, and powders may contain from about 1 to about 95 wt% of the compound of the present invention, preferably from about 15 to about 90 wt% of the compound of the present invention, based on the total weight of the composition. An example of a tablet composition may contain, for example, up to about 80 wt% of the active pharmaceutical ingredient, from about 10 wt% to about 90 wt% of a binder, from about 0 wt% to about 85 wt% of a diluent, from about 2 wt% to about 10 wt% of a disintegrant, and from about 0.25 wt% to about 10 wt% of a lubricant.

[0080] When administered in liquid or suspension form, a functional liquid and / or liquid carrier such as water, petroleum, or animal or vegetable oil may be added. Liquid compositions may further contain saline, sugar alcohol solutions, dextrose or other sugar solutions, or glycols. When administered in liquid or suspension form, the composition may contain from about 0.5% to about 90% by weight of the compound of the present invention, preferably from about 1% to about 50% by weight of the compound of the present invention. In one contemplated embodiment, the liquid carrier is non-aqueous or substantially non-aqueous. For administration in liquid form, the composition may be supplied as a rapidly dissolving solid formulation for dissolution or suspension immediately prior to administration.

[0081] When the pharmaceutical composition of the present invention is administered intravenously, transdermally, or subcutaneously, it is in the form of a parenteral aqueous solution that does not contain a pyrogen. The preparation of such parenteral solutions, taking into account pH, isotonicity, stability, etc., is within the skill of the art. Preferred compositions for intravenous, transdermal, or subcutaneous injection generally contain an isotonic vehicle. Such compositions can be prepared for administration as a solution of the free base or a pharmacologically acceptable salt in water suitably mixed with a surfactant such as hydroxypropyl cellulose. Dispersions in glycerol, liquid polyethylene glycol, and mixtures thereof, as well as dispersions in oils, can also be prepared. Under normal storage and use conditions, such preparations may optionally contain a preservative to prevent the growth of microorganisms.

[0082] Injectable compositions may include sterile aqueous solutions, suspensions, or dispersions, and sterile powders for the extemporaneous preparation of sterile injectable solutions, suspensions, or dispersions. Sterile injectable solutions are prepared by incorporating the active compound in the required amount in an appropriate solvent with various other ingredients, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the various sterile active ingredients into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above. In embodiments of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze-drying techniques, which produce a powder of the active ingredient and any additional required ingredients from a previously sterile-filtered solution thereof.

[0083] To achieve controlled release of the active compound upon contact with body fluids in the gastrointestinal tract and to provide substantially constant and effective levels of the active compound in the plasma, sustained-release or sustained-release formulations may be prepared. For example, release may be controlled by one or more of dissolution, diffusion, and ion exchange. Furthermore, sustained-release approaches may enhance absorption through saturation or restriction pathways within the gastrointestinal tract. For example, for this purpose, the compound may be embedded in a polymer matrix comprising a biodegradable polymer, a water-soluble polymer, or a mixture thereof, and optionally a suitable surfactant. Embedding in this context may mean incorporating microparticles into the polymer matrix. Controlled-release formulations may also be obtained by encapsulating dispersed microparticles or emulsified microdroplets using known dispersion or emulsion coating techniques.

[0084] The pharmaceutical composition of the present invention may be formulated for parenteral administration by injection (e.g., bolus injection or continuous infusion). The injectable formulation may be presented in unit dosage form (e.g., ampoules or multi-dose containers) with an added preservative. The composition may take the form of a suspension, solution, or emulsion in an oily or aqueous vehicle, and may contain formulatory agents such as suspending, stabilizing, and / or dispersing agents.

[0085] The pharmaceutical composition may be presented in unit dosage form suitable for single administration of precise dosages.

[0086] A pharmaceutical composition containing the compound of the present invention can be used according to the method described below.

[0087] Treatment methods

[0088] In another aspect, the present invention provides a method for treating a disease requiring inhibition of SOS1 activity. The method comprises administering a therapeutically effective amount of a compound of the present invention, such as a compound defined by Formula 1, or a pharmaceutical composition comprising such a compound of the present invention, to a patient in need of inhibition of SOS1 activity.

[0089] Diseases that require inhibition of SOS1 activity include cancer.

[0090] In one embodiment, the present invention provides a method for treating cancer. The method comprises administering to a patient with cancer a therapeutically effective amount of a compound of the present invention, such as a compound defined by Formula 1, or a pharmaceutical composition comprising such a compound of the present invention. In one specific embodiment, the cancer is a KRAS G12C, G12D, G12V, or G13D mutant cancer.

[0091] In a method for treating cancer, the compound of the present invention may be administered alone or in combination with at least one other drug. These other drugs and the compound of the present invention may be administered simultaneously or sequentially. Some examples of drugs that may be administered in combination with the drug of the present invention include RAS inhibitors, MEK inhibitors, and EGFR inhibitors. Specific examples of RAS inhibitors include sotorasib and adagrasib. Specific examples of MEK inhibitors include trametinib, cobimetinib, selumetinib, and binimetinib. Specific examples of EGFR inhibitors include gefitinib, erlotinib, afatinib, brigatinib, osimertinib, cetuximab, and panitumumab.

[0092] As used herein, unless specifically stated otherwise, a “therapeutically effective amount” of a compound refers to an amount of the compound sufficient to delay or minimize one or more symptoms associated with a disease, disorder, or condition, or to provide a therapeutic effect for a disease, disorder, or condition. The term “therapeutically effective amount” encompasses an amount that improves overall healing, an amount that alleviates or avoids a symptom or cause of a disease or condition, and an amount that enhances the therapeutic effectiveness of another therapeutic agent.

[0093] The amount of compound administered may vary depending on the subject being treated, the subject's age, health, sex, and body weight, the type (if any) of concurrent treatment, the severity of the pain, the nature of the desired effect, the treatment regimen and frequency, and the prescribing physician's judgment. The frequency of administration may also vary depending on the pharmacodynamic effects on arterial oxygen pressure. However, the most desirable dosage can be tailored to the individual subject, as understood and determined by those skilled in the art without undue experimentation. This typically involves adjusting the standard dose (e.g., reducing the dose if the patient is underweight).

[0094] Although individual case needs may vary, determining the optimal range of therapeutically effective doses of a compound is common sense in this field. When administered to humans in the therapeutic or preventive treatment of conditions and disorders using the compounds of the present invention, for example, a typical dosage of the compounds of the present invention may be from about 0.05 mg / kg / day to about 50 mg / kg / day, for example, at least 0.05 mg / kg, at least 0.08 mg / kg, at least 0.1 mg / kg, at least 0.2 mg / kg, at least 0.3 mg / kg, at least 0.4 mg / kg, or at least 0.5 mg / kg, preferably not more than 50 mg / kg, not more than 40 mg / kg, not more than 30 mg / kg, not more than 20 mg / kg, or not more than 10 mg / kg, which may be, for example, from about 2.5 mg / day (0.5 mg / kg×5 kg) to about 5000 mg / day (50 mg / kg×100 kg). For example, the dosage of the compound may be from about 0.1 mg / kg / day to about 50 mg / kg / day, from about 0.05 mg / kg / day to about 10 mg / kg / day, from about 0.05 mg / kg / day to about 5 mg / kg / day, from about 0.05 mg / kg / day to about 3 mg / kg / day, from about 0.07 mg / kg / day to about 3 mg / kg / day, from about 0.09 mg / kg / day to about 3 mg / kg / day, from about 0.05 mg / kg / day to about 0.1 mg / kg / day, from about 0.1 mg / kg / day to about 1 mg / kg / day, from about 1 mg / kg / day to about 10 mg / kg / day, from about 1 mg / kg / day to about 5 mg / kg / day, from about 1 mg / kg / day to about 3 mg / kg / day, from about 3 mg / day to about The dose may be 500 mg / day, about 5 mg / day to about 250 mg / day, about 10 mg / day to about 100 mg / day, about 3 mg / day to about 10 mg / day, or about 100 mg / day to about 250 mg / day. This dose may be administered as a single dose or divided into multiple doses.

[0095] In another aspect of the present invention, a method for inhibiting intracellular SOS1 activity is provided. The method comprises contacting a cell in which inhibition of SOS1 activity is desired with an effective amount of a compound of the present invention, e.g., a compound defined by Formula 1.

[0096] [Example]

[0097] Hereinafter, the present invention will be described in more detail through the following examples and experimental examples. However, these examples and experimental examples are intended only to aid understanding of the present invention and are not intended to limit the scope of the present invention in any way. Various changes and modifications may be made to the present examples, and such changes and modifications also fall within the scope of the appended claims.

[0098] The full names of the abbreviations described in the synthesis method below are as follows.

[0099] Ph: Phenyl TEA: Triethylamine THF: Tetrahydrofuran

[0100] Pd2(dba)3: Tris(dibenzylideneacetone)dipalladium

[0101] XantPhos: (9,9-dimethyl-9H-xanthene-4,5-diyl)bis(diphenylphosphane)

[0102]

[0103] [Example 1] Preparation of (R)-6,7-dimethoxy-2-methyl-N-(1-(4-(2-((methylamino)methyl)phenyl)selenophen-2-yl)ethyl)quinazolin-4-amine

[0104]

[0105] The compound of Example 1 was prepared by the reaction scheme below.

[0106]

[0107] Step (1) Preparation of benzyl (2-bromobenzyl)(methyl)carbamate

[0108]

[0109] 1-(2-Bromophenyl)-N-methylmethanamine (5.00 g, 24.9 mmol, 1.00 eq) was dissolved in dichloromethane (50.0 mL), and then triethylamine (5.06 g, 49.9 mmol, 6.96 mL, 2.00 eq) and benzyl carbonochloridate (4.69 g, 27.4 mmol, 3.92 mL, 1.10 eq) were added at 0°C. The mixture was stirred at 20°C for 2 h. At 0°C, triethylamine (2.53 g, 24.9 mmol, 3.48 mL, 1.00 eq) and benzyl carbonochloridate (2.13 g, 12.5 mmol, 1.78 mL, 0.500 eq) were added to the mixture, and the mixture was stirred at 20°C for 12 h. The mixture was concentrated under reduced pressure to obtain a residue, which was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1) to give benzyl (2-bromobenzyl)(methyl)carbamate (7.80 g, 23.3 mmol, 93% yield).

[0110] 1 H NMR (400 MHz, CDCl3)δ= 7.56 (d,J= 7.2 Hz, 1H), 7.45 - 7.20 (m, 7H), 7.17 - 7.12 (m, 1H), 5.20 (br d,J= 16.4 Hz, 2H), 4.62 (br d,J= 18.4 Hz, 2H), 2.96 (br d,J= 11.6 Hz, 3H).

[0111] Step (2) Preparation of benzyl methyl(2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)carbamate.

[0112]

[0113] Benzyl (2-bromobenzyl)(methyl)carbamate (2.00 g, 5.98 mmol, 1.00 eq) was dissolved in dioxane (30.0 mL), and 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxoborolane) (2.28 g, 8.98 mmol, 1.50 eq), potassium acetate (1.47 g, 14.9 mmol, 2.50 eq), and [1,1-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (437 mg, 598 μmol, 0.100 eq) were added. The mixture was stirred at 110°C for 12 h under a nitrogen atmosphere. The mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = from 100 / 1 to 0 / 1) to obtain benzyl methyl(2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)carbamate (2.40 g, 5.67 mmol, 94% yield, 90% purity) as a green oil.

[0114] 1 H NMR (400 MHz, CDCl3)δ= 7.83 (br d,J= 7.2 Hz, 1H), 7.45 - 7.27 (m, 7H), 7.26 - 7.19 (m, 1H), 5.25 - 5.14 (m, 2H), 4.86 (br d,J= 8.8 Hz, 2H), 2.90 (br d,J= 12.8 Hz, 3H), 1.34 (s, 12H).

[0115] Step (3) Preparation of benzyl (2-(5-acetylselenophen-3-yl)benzyl)(methyl)carbabate

[0116]

[0117] 1-(4-Bromoselenophen-2-yl)ethan-1-one (1.10 g, 4.37 mmol, 1.00 eq) and benzyl methyl(2-(4,4,5,5-tetramethyl-1,3,2-dioxoborolan-2-yl)benzyl)carbamate (2.22 g, 5.24 mmol, 1.20 eq) were dissolved in dioxane (15.0 mL) and water (3.00 mL), and then [1,1-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (319 mg, 436 μmol, 0.100 eq) and potassium carbonate (1.81 g, 13.1 mmol, 3.00 eq) were added. The mixture was stirred at 100 °C for 2 h under a nitrogen atmosphere. The mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = from 100 / 1 to 0 / 1) and then lyophilized to obtain benzyl (2-(5-acetylselenophen-3-yl)benzyl)(methyl)carbamate (1.15 g, 2.70 mmol, 61% yield) as a pale yellow oil.

[0118] 1 H NMR (400 MHz, CDCl3)δ= 8.27 - 7.97 (m, 1H), 7.96 - 7.74 (m, 1H), 7.42 - 7.28 (m, 8H), 7.25 - 7.19 (m, 1H), 5.20 - 5.01 (m, 2H), 4.62 - 4.42 (m, 2H), 2.89 - 2.71 (m, 3H), 2.64 - 2.46 (m, 3H).

[0119] Step (4) Preparation of benzyl (R,E)-(2-(5-(1-((tert-butylsulfinyl)imino)ethyl)selenophen-3-yl)benzyl)(methyl)carbamate

[0120]

[0121] Benzyl (2-(5-acetylselenophen-3-yl)benzyl)(methyl)carbamate (300 mg, 703 μmol, 1.00 eq) was dissolved in tetrahydrofuran (6.00 mL), and then (R)-2-methylpropane-2-sulfinylamide (255 mg, 2.11 mmol, 3.00 eq) and titanium(IV) ethoxide (481 mg, 2.11 mmol, 437 μL, 3.00 eq) were added. The mixture was stirred at 70 °C for 12 h. The mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate=100 / 1 to 0 / 1) to give benzyl (R,E)-(2-(5-(1-((tert-butylsulfinyl)imino)ethyl)selenophen-3-yl)benzyl)(methyl)carbamate (270 mg, 489 μmol, 69% yield, 96% purity) as a yellow oil.

[0122] 1 H NMR (400 MHz, CDCl3)δ= 8.04 - 7.86 (m, 1H), 7.79 - 7.54 (m, 1H), 7.42 - 7.27 (m, 8H), 7.25 - 7.20 (m, 1H), 5.12 (br d,J= 17.2 Hz, 2H), 4.62 - 4.44 (m, 2H), 2.92 - 2.63 (m, 6H), 1.32 (s, 9H).

[0123] Step (5) Preparation of benzyl (2-(5-(1-(((R)-tert-butylsulfinyl)amino)ethyl)selenophen-3-yl)benzyl)(methyl)carbamate

[0124]

[0125] Benzyl (R,E)-(2-(5-(1-((tert-butylsulfinyl)imino)ethyl)selenophene-3-yl)benzyl)(methyl)carbamate (270 mg, 509 μmol, 1.00 eq) was dissolved in tetrahydrofuran (4.00 mL), and then sodium borohydride (57.8 mg, 1.53 mmol, 3.00 eq) was added at 0 °C. The mixture was stirred at 25 °C for 2 h. The mixture was poured into ice water (10 mL), extracted with ethyl acetate (3 × 10 mL), washed with brine (20 mL), and dried over anhydrous sodium sulfate. The residue was then filtered and concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1) to give benzyl (2-(5-(1-(((R)-tert-butylsulfinyl)amino)ethyl)selenophen-3-yl)benzyl)(methyl)carbamate (220 mg, 393 μmol, 77% yield, 95% purity) as a colorless oil.

[0126] 1 H NMR (400 MHz, CDCl3)δ= 7.72 - 7.62 (m, 1H), 7.40 - 7.27 (m, 8H), 7.25 - 7.12 (m, 2H), 5.14 (br d,J= 8.0 Hz, 2H), 4.84 (br s, 1H), 4.63 - 4.47 (m, 2H), 2.90 - 2.67 (m, 3H), 1.61 (br d,J= 6.4 Hz, 3H), 1.27 (s, 9H).

[0127] Step (6) Preparation of benzyl (R)-(2-(5-(1-aminoethyl)selenophene-3-yl)benzyl)(methyl)carbamate

[0128]

[0129] Benzyl (R)-(2-(5-(1-aminoethyl)selenophene-3-yl)benzyl)(methyl)carbamate (220 mg, 351 μmol, 1.00 eq) was dissolved in methanol (2.00 mL), and then concentrated hydrochloric acid (1.00 mL) was added. The mixture was stirred at 25 °C for 2 h. The pH of the mixture was adjusted to 7 by adding sodium hydroxide solution (2 N) at 0 °C, extracted with a mixture of dichloromethane and isopropanol (3:1) (3 × 20 mL), and dried over anhydrous sodium sulfate. After filtration, the mixture was concentrated under reduced pressure to obtain benzyl (R)-(2-(5-(1-aminoethyl)selenophene-3-yl)benzyl)(methyl)carbamate (200 mg, raw material) as a yellow oil.

[0130] 1 H NMR (400 MHz, DMSO-d6)δ= 8.08 - 7.91 (m, 1H), 7.44 - 7.24 (m, 9H), 7.17 (br d,J= 7.6 Hz, 3H), 5.17 - 4.97 (m, 2H), 4.65 - 4.54 (m, 1H), 4.50 (s, 2H), 2.77 (s, 3H), 1.51 (br s, 3H).

[0131] Step (7) Preparation of benzyl (R)-(2-(5-(1-((6,7-dimethoxy-2-methylquinazolin-4-yl)amino)ethyl)selenophen-3-yl)benzyl)(methyl)carbamate

[0132]

[0133] Benzyl (R)-(2-(5-(1-aminoethyl)selenophene-3-yl)benzyl)(methyl)carbamate (75.0 mg, 175 μmol, 1.00 eq) was dissolved in dimethylformamide (1.50 mL), and then 4-chloro-6,7-dimethoxy-2-methylquinazoline (41.8 mg, 175 μmol, 1.00 eq), N,N-diisopropylethylamine (68.0 mg, 526 μmol, 91.7 μL, 3.00 eq), and 4A MS (225 mg) were added. The mixture was stirred at 20 °C for 0.5 h. The mixture was then stirred at 130 °C for 3.5 h. The mixture was filtered, and the filtrate was purified by Prep-HPLC (Column: Waters Xbridge 150×25mm×5 μm; Mobile phase: [water(zinc carbonate)-acetonitrile]; Gradient: 58%–88% B, for 9 min) to obtain benzyl (R)-(2-(5-(1-((6,7-Imethoxy-2-methylquinazolin-4-yl)amino)ethyl)selenophen-3-yl)benzyl)(methyl)carbamate (74.0 mg, 116 μmol, 33% yield, 99% purity) as a yellow solid.

[0134] 1 H NMR (400 MHz, DMSO-d6)δ= 8.12 (br d,J= 7.6 Hz, 1H), 7.89 - 7.77 (m, 1H), 7.62 (s, 1H), 7.40 - 7.24 (m, 8H), 7.16 (br d,J= 7.2 Hz, 2H), 7.05 (s, 1H), 5.95 - 5.78 (m, 1H), 5.14 - 4.96 (m, 2H), 4.49 (s, 2H), 3.87 (d,J= 4.0 Hz, 6H), 2.75 (br s, 3H), 2.44 (s, 3H), 1.72 (br s, 3H).

[0135] Step (8) Preparation of (R)-6,7-dimethoxy-2-methyl-N-(1-(4-(2-((methylamino)methyl)phenyl)selenophen-2-yl)ethyl)quinazolin-4-amine

[0136]

[0137] Benzyl (R)-(2-(5-(1-((6,7-Imethoxy-2-methylquinazolin-4-yl)amino)ethyl)selenophene-3-yl)benzyl)(methyl)carbamate (40 mg, 63.5 μmol, 1.00 eq) was dissolved in dichloromethane (1.00 mL), and then iodotrimethylsilane (38.1 mg, 191 μmol, 25.9 μL, 3.00 eq) was added. The mixture was stirred at 25 °C for 12 h. Then, 2 mL of methanol was added, and the resulting mixture was stirred at room temperature for an additional 30 min and concentrated under reduced pressure. The residue was purified by Prep-HPLC (Column: Waters Xbridge 150×25mm×5μm; Mobile phase: [water(zinc carbonate)-acetonitrile]; Gradient: 32%–62% B, for 9 min) to afford (R)-6,7-dimethoxy-2-methyl-N-(1-(4-(2-((methylamino)methyl)phenyl)selenophen-2-yl)ethyl)quinazolin-4-amine (11.0 mg, 22.1 μmol, 35% yield, 99% purity) as a white solid.

[0138] 1 H NMR (400 MHz, DMSO-d6) δ = 8.15 (d,J= 8.1 Hz, 1H), 7.97 (d,J= 1.3 Hz, 1H), 7.65 (s, 1H), 7.49 - 7.42 (m, 2H), 7.33 - 7.23 (m, 3H), 7.06 (s, 1H), 5.91 (m, 1H), 3.88 (s, 6H), 3.57 (s, 2H), 2.46 (s, 3H), 2.23 (s, 3H), 1.74 (d,J= 7.0 Hz, 3H).

[0139]

[0140] [Example 2] Preparation of 7-methoxy-2-methyl-N-((R)-1-(4-(2-((methylamino)methyl)phenyl)selenophen-2-yl)ethyl)-6-(((S)-tetrahydrofuran-3-yl)oxy)quinazolin-4-amine.

[0141] Compound 2 was prepared by the reaction scheme below.

[0142] [Correction pursuant to Rule 91, 04.09.2024]

[0143] Step (1) Preparation of 7-methoxy-2-methylquinazoline-4,6-diol

[0144]

[0145] 6,7-Dimethoxy-2-methylquinazolin-4-ol (4.80 g, 21.8 mmol, 1.00 eq) was dissolved in methanesulfonic acid (25.0 mL), and DL-methionine (4.88 g, 32.7 mmol, 1.50 eq) was added. The mixture was stirred at 120°C for 36 h. The reaction mixture was poured into ice water (500 mL) and basified with soda water until the pH became 9. The solution was filtered to obtain a filter cake. The filter cake was triturated with 3 M hydrochloric acid at 25°C for 12 h. The obtained mixture was filtered, and the filter cake was dried under vacuum to obtain 7-methoxy-2-methylquinazoline-4,6-diol (2.40 g, 11.6 mmol, 53% yield) as a yellow solid.

[0146] 1 H NMR (400 MHz, DMSO-d6) δ= 9.67 (s, 1H), 7.32 (s, 1H), 7.01 (s, 1H), 3.87 (s, 3H), 2.28 (s, 3H).

[0147] Step (2) Preparation of (S)-7-methoxy-2-methyl-6-((tetrahydrofuran-3-yl)oxy)quinazolin-4-ol

[0148]

[0149] 7-Methoxy-2-methylquinazoline-4,6-diol (1.20 g, 5.82 mmol, 1.00 eq) was dissolved in dimethylformamide (50.0 mL), and (R)-tetrahydrofuran-3-yl 4-methylbenzenesulfonate (1.41 g, 5.82 mmol, 1.00 eq) and cesium carbonate (2.84 g, 8.73 mmol, 1.50 eq) were added. The mixture was stirred at 25°C for 12 h. The reaction mixture was filtered to obtain a filtrate. The filtrate was concentrated in vacuo to obtain a residue. The residue was purified by Prep-HPLC (column: Waters Xbridge Prep OBD C18 150×40 mm×10 μm; mobile phase: [water(ammonium sulfite)-acetonitrile]; gradient: 0%–30% B for 15 min) to give (S)-7-methoxy-2-methyl-6-((tetrahydrofuran-3-yl)oxy)quinazolin-4-ol (800 mg, 2.90 mmol, 49.8% yield) as a white solid.

[0150] 1 H NMR (400 MHz, DMSO-d6)δ= 7.34 (s, 1H), 7.04 (s, 1H), 5.08 (br d,J= 1.2 Hz, 1H), 3.91 - 3.73 (m, 7H), 2.29 (s, 3H), 2.27 - 2.18 (m, 1H), 2.04 - 1.95 (m, 1H).

[0151] Step (3) Preparation of benzyl (2-(5-((R)-1-((7-methoxy-2-methyl-6-(((S)-tetrahydrofuran-3-yl)oxy)quinazolin-4-yl)amino)ethyl) selenophen-3-yl)benzyl)(methyl)carbamate

[0152]

[0153] (S)-7-Methoxy-2-methyl-6-((tetrahydrofuran-3-yl)oxy)quinazolin-4-ol (200 mg, 724 μmol, 1.00 eq), ((1H-benzo[d][1,2,3]triazol-1-yl)oxy)tris(dimethylamino)phosphonium hexafluorophosphate (V) (416 mg, 941 μmol, 1.30 eq), and 2,3,4,6,7,8,9,10-octahydropyrimido[1,2-a]azepine (548 mg, 3.60 mmol, 543 μL, 4.97 eq) were dissolved in dimethylformamide (10.0 mL) and stirred at 25°C for 1 h. Afterwards, benzyl (R)-(2-(5-(1-aminoethyl)selenophene-3-yl)benzyl)(methyl)carbamate (309 mg, 724 μmol, 1.00 eq) was added, and the mixture was stirred at 25°C for 12 h. The mixture was concentrated in vacuo to obtain a residue. The residue was purified by Prep-HPLC (column: YMC-Actus Triart C18 150×30 mm×7 μm; mobile phase: [water(formic acid)-acetonitrile]; gradient: 33%–63% B for 10 min) to give benzyl (2-(5-((R)-1-((7-methoxy-2-methyl-6-(((S)-tetrahydrofuran-3-yl)oxy)quinazolin-4-yl)amino)ethyl)selenophen-3-yl)benzyl)(methyl)carbamate (54.0 mg, 78.8 μmol, 10.9% yield) as a yellow solid.

[0154] 1H NMR (400 MHz, DMSO-d6)δ= 14.17 - 13.92 (m, 1H), 9.80 (br dd,J= 2.4, 3.6 Hz, 1H), 8.03 - 7.93 (m, 1H), 7.39 - 7.23 (m, 8H), 7.18 (br d,J= 7.2 Hz, 1H), 7.14 - 7.05 (m, 2H), 6.12 - 6.00 (m, 1H), 5.18 - 5.12 (m, 1H), 5.08 - 4.93 (m, 2H), 4.49 (s, 2H), 3.97 (s, 4H), 3.87 - 3.77 (m, 3H), 2.76 (s, 3H), 2.63 (br s, 3H), 2.03 - 1.96 (m, 2H), 1.78 (br s, 3H).

[0155] Step (4) Preparation of 7-methoxy-2-methyl-N-((R)-1-(4-(2-((methylamino)methyl)phenyl)selenophen-2-yl)ethyl)-6-(((S)-tetrahydrofuran-3-yl)oxy)quinazolin-4-amine

[0156]

[0157] Benzyl (2-(5-((R)-1-((7-methoxy-2-methyl-6-(((S)-tetrahydrofuran-3-yl)oxy)quinazolin-4-yl)amino)ethyl)selenophene-3-yl)benzyl)(methyl)carbamate (45.0 mg, 65.6 μmol, 1.00 eq) was dissolved in dimethyl sulfoxide (2.00 mL), followed by addition of sodium hydroxide (13.1 mg, 328 μmol, 5.00 eq). The mixture was stirred at 70°C for 6 h. The mixture was purified by Prep-HPLC (column: YMC-Actus Triart C18 150×30 mm×7 μm; mobile phase: [water(formic acid)-acetonitrile]; gradient: 8%–38% B for 10 min) and lyophilized to obtain 7-methoxy-2-methyl-N-((R)-1-(4-(2-((methylamino)methyl)phenyl)selenophen-2-yl)ethyl)-6-(((S)-tetrahydrofuran-3-yl)oxy)quinazolin-4-amine (10.56 mg, 18.27 μmol, 27.8% yield, 95.4% purity) as a brown solid.

[0158] 1 H NMR (400 MHz, DMSO-d6)δ= 8.27 (s, 1H), 8.19 (br d,J= 8.4 Hz, 1H), 7.95 (s, 1H), 7.67 - 7.61 (m, 1H), 7.54 (br d,J= 6.4 Hz, 1H), 7.40 (s, 1H), 7.37 - 7.29 (m, 3H), 7.07 (s, 1H), 5.90 (br t,J= 7.2 Hz, 1H), 5.20 - 5.08 (m, 1H), 3.96 (br dd,J= 4.4, 10.0 Hz, 1H), 3.87 (s, 3H), 3.84 - 3.75 (m, 6H), 2.45 (s, 3H), 2.34 (s, 3H), 2.29 (br s, 1H), 1.97 (br s, 1H), 1.74 (d,J= 6.8 Hz, 3H).

[0159]

[0160] [Example 3] Preparation of (R)-(2-chloro-4-((1-(4-(2-((methylamino)methyl)phenyl)selenophene-2-yl)ethyl)amino)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(4-methoxytetrahydro-2H-pyran-4-yl)methanone.

[0161] The compound of Example 3 was prepared by the reaction scheme below.

[0162]

[0163] Step (1) Preparation of tert-butyl (R)-4-((1-(4-(2-((((benzyloxy)carbonyl)(methyl)amino)methyl)phenyl)selenophene-2-yl)ethyl)amino)-2-chloro-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate.

[0164]

[0165] tert-Butyl 2,4-dichloro-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (100 mg, 344 μmol, 1.00 eq) and benzyl (R)-(2-(5-(1-aminoethyl)selenophene-3-yl)benzyl)(methyl)carbamate (147 mg, 344 μmol, 1.00 eq) were dissolved in tetrahydrofuran (1.00 mL), and N,N-diisopropylethylamine (133 mg, 1.03 mmol, 180 μL, 3.00 eq) was added. The mixture was stirred at 40 °C for 12 h. The mixture was concentrated in vacuo to obtain a residue. The residue was purified by Prep-HPLC (column: YMC-Actus Triart C18 150×30 mm×7 μm; mobile phase: [water(formic acid)-acetonitrile]; gradient: 80%–100% B for 10 min) and lyophilized to obtain tert-butyl (R)-4-((1-(4-(2-((((benzyloxy)carbonyl)(methyl)amino)methyl)phenyl)selenophen-2-yl)ethyl)amino)-2-chloro-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (140 mg, 203 μmol, 59% yield, 99% purity) as a yellow solid.

[0166] 1 H NMR (400 MHz, DMSO-d6)δ= 8.30 (br d,J= 5.6 Hz, 1H), 7.89 (br d,J= 17.6 Hz, 1H), 7.43 - 7.24 (m, 8H), 7.23 - 7.10 (m, 2H), 5.60 - 5.47 (m, 1H), 5.14 - 4.95 (m, 2H), 4.48 (br s, 2H), 4.36 (br d,J= 5.6 Hz, 4H), 2.76 (br s, 3H), 1.59 (br s, 3H), 1.45 (s, 9H).

[0167] Step (2) Preparation of benzyl (R)-(2-(5-(1-((2-chloro-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)amino)ethyl)selenophen-3-yl)benzyl)(methyl)carbamate

[0168]

[0169] tert-Butyl (R)-4-((1-(4-(2-(((benzyloxy)carbonyl)(methyl)amino)methyl)phenyl)selenophene-2-yl)ethyl)amino)-2-chloro-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (120 mg, 176 μmol, 1.00 eq) was dissolved in dichloromethane (1.20 mL) and trifluoroacetic acid (0.400 mL). The mixture was stirred at 20 °C for 2 h. The mixture was concentrated in vacuo to obtain the target compound (R)-(2-(5-(1-((2-chloro-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)amino)ethyl)selenophen-3-yl)benzyl)(methyl)carbamate (102 mg) as a purple oil.

[0170] MS (ESI) m / z 582.2 [M+H] +

[0171] Step (3) Preparation of benzyl (R)-(2-(5-(1-((2-chloro-6-(4-methoxytetrahydro-2H-pyran-4-carbonyl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)amino)ethyl)selenophene-3-yl)benzyl)(methyl)carbamate

[0172]

[0173] 4-Methoxytetrahydro-2H-pyran-4-carboxylic acid (28.1 mg, 175 μmol, 1.00 eq) was dissolved in dimethylformamide (1.00 mL). 2-(3H-[1,2,3]-triazolo[4,5-b]pyridin-3-yl)-1,1,3,3-tetramethylisourea hexafluorophosphate (V) (100 mg, 263 μmol, 1.50 eq) and triethylamine (53.3 mg, 526 μmol, 73.3 μL, 3.00 eq) were added. The mixture was stirred at 25 °C for 0.5 h. Then, benzyl (R)-(2-(5-(1-((2-chloro-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)amino)ethyl)selenophene-3-yl)benzyl)(methyl)carbamate (102 mg, 175 μmol, 1.00 eq) and triethylamine (53.3 mg, 526 μmol, 73.3 μL, 3.00 eq) were added to the mixture. The mixture was stirred at 25 °C for 2 h. After filtering the mixture, the filtrate was purified by Prep-HPLC (column: YMC-Actus Triart C18 150×30 mm×7 μm; mobile phase: [water (formic acid)-acetonitrile]; gradient: 63%–93% B for 10 min) and lyophilized to obtain benzyl (R)-(2-(5-(1-((2-chloro-6-(4methoxytetrahydro-2H-pyran-4-carbonyl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)amino)ethyl)selenophene-3-yl)benzyl)(methyl)carbamate (100 mg, 136 μmol, 77% yield, 99% purity) as a white solid.

[0174] 1H NMR (400 MHz, DMSO-d6)δ= 8.41 (br dd,J= 3.2, 3.6 Hz, 1H), 8.00 - 7.81 (m, 1H), 7.40 - 7.22 (m, 8H), 7.21 - 7.09 (m, 2H), 5.61 - 5.47 (m, 1H), 5.14 - 4.94 (m, 2H), 4.78 (br s, 2H), 4.50 (br s, 4H), 3.75 - 3.51 (m, 4H), 3.17 - 3.11 (m, 3H), 2.76 (br s, 3H), 2.02 - 1.76 (m, 4H), 1.60 (br d,J= 2.4 Hz, 3H).

[0175] Step (4) Preparation of (R)-(2-chloro-4-((1-(4-(2-((methylamino)methyl)phenyl)selenophen-2-yl)ethyl)amino)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(4-methoxytetrahydro-2H-pyran-4-yl)methanone

[0176]

[0177] Benzyl (R)-(2-(5-(1-((2-chloro-6-(4-methoxytetrahydro-2H-pyran-4-carbonyl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)amino)ethyl)selenophene-3-yl)benzyl)(methyl)carbamate (90.0 mg, 124 μmol, 1.00 eq) was dissolved in dichloromethane (1.00 mL), followed by the addition of iodotrimethylsilane (29.8 mg, 149 μmol, 20.3 μL, 1.20 eq). The mixture was stirred at 25 °C for 1 h. The mixture was concentrated in vacuo to obtain the residue, which was purified by Prep-HPLC (column: YMC-Actus Triart C18 150×30 mm×7 μm; mobile phase: [water(formic acid)-acetonitrile]; gradient: 20%–50% B for 10 min) and lyophilized to obtain (R)-(2-chloro-4-((1-(4-(2-((methylamino)methyl)phenyl)selenophen-2-yl)ethyl)amino)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(4-methoxytetrahydro-2H-pyran-4-yl)methanone (25.46 mg, 32.9 μmol, 26.4% yield, 96.8% purity, formate) as a white solid.

[0178] 1 H NMR (400 MHz, DMSO-d6)δ= 8.48 - 8.37 (m, 1H), 8.21 (s, 1H), 8.05 - 7.98 (m, 1H), 7.52 - 7.47 (m, 1H), 7.41 - 7.28 (m, 4H), 5.64 - 5.51 (m, 1H), 4.80 (br s, 2H), 4.53 (br s, 2H), 3.71 (br s, 2H), 3.57 (br s, 4H), 3.17 - 3.13 (m, 3H), 2.32 (s, 3H), 1.97 - 1.81 (m, 4H), 1.66 - 1.58 (m, 3H).

[0179]

[0180] [Example 4] Preparation of (R)-4-methyl-N-(1-(4-(2-((methylamino)methyl)phenyl)selenophen-2-yl)ethyl)-7-morpholinopyrido[3,4-d]pyridazin-1-amine

[0181] The compound of Example 4 was prepared by the reaction scheme below.

[0182]

[0183] Step (1) Preparation of benzyl (R)-(2-(5-(1-((7-chloro-4-methylpyrido[3,4-d]pyridazin-1-yl)amino)ethyl)selenophen-3-yl)benzyl)(methyl)carbamate

[0184]

[0185] 1,7-Dichloro-4-methylpyrido[3,4-d]pyridazine (0.400 g, 1.87 mmol, 1.00 eq) and benzyl (R)-(2-(5-(1-aminoethyl)selenophene-3-yl)benzyl)(methyl)carbamate (798 mg, 1.87 mmol, 1.00 eq) were dissolved in dimethyl sulfoxide (3.00 mL). Then, potassium fluoride (325 mg, 5.61 mmol, 3.00 eq) and N,N-diisopropylethylamine (724 mg, 5.61 mmol, 976 μL, 3.00 eq) were added. The mixture was stirred at 130 °C for 12 h. The reaction mixture was quenched by adding water (10 mL), and then extracted with dichloromethane (2 × 15 mL). The combined organic layers were washed with aqueous sodium chloride solution and dried over anhydrous sodium sulfate. After filtration, the residue was concentrated in vacuo to obtain a residue. The residue was purified by Prep-HPLC (column: Phenomenex luna C18 150 × 40 mm × 15 μm; mobile phase: [water (formic acid)-acetonitrile]; gradient: 38%–68% B for 15 min) and lyophilized to obtain benzyl (R)-(2-(5-(1-((7-chloro-4-methylpyrido[3,4-d]pyridazin-1-yl)amino)ethyl)selenophene-3-yl)benzyl)(methyl)carbamate (268 mg, 443 μmol, 23.7% yield) as a yellow solid.

[0186] 1 H NMR (400 MHz, DMSO-d6)δ= 9.32 (s, 1H), 8.55 (s, 1H), 7.40 - 7.26 (m, 10H), 7.21 - 7.15 (m, 2H), 5.84 (br t,J= 6.8 Hz, 1H), 5.12 - 5.01 (m, 2H), 4.50 (br s, 2H), 2.79 (s, 6H), 1.74 (br s, 3H).

[0187] Step (2) Preparation of benzyl (R)-methyl(2-(5-(1-((4-methyl-7-morpholinopyrido[3,4-d]pyridazin-1-yl)amino)ethyl)selenophen-3-yl)benzyl)carbamate

[0188]

[0189] Morpholine (193 mg, 2.21 mmol, 195 μL, 5.00 eq) and benzyl (R)-(2-(5-(1-((7-chloro-4-methylpyrido[3,4-d]pyridazin-1-yl)amino)ethyl)selenophene-3-yl)benzyl)(methyl)carbamate (268 mg, 442 μmol, 1.00 eq) were dissolved in dimethyl sulfoxide (3.00 mL), to which potassium fluoride (100 mg, 664 μmol, 24.5 μL, 1.50 eq) and diisopropylethylamine (171 mg, 1.33 mmol, 231 μL, 3.00 eq) were added. The mixture was stirred at 130 °C for 12 h, and then water (15 mL) was added. Subsequently, after extraction with dichloromethane (2 × 10 mL), the organic solvent was washed with saturated sodium chloride solution (2 × 10 mL). The organic solvent was dried over anhydrous sodium sulfate, filtered, and distilled under reduced pressure to obtain a residue. The residue was purified by Prep-HPLC (column: YMC-Actus Triart C18 150 × 30 mm × 7 μm; mobile phase: [water (formic acid)-acetonitrile; gradient: 35%–65% B for 10 min)] and lyophilized to obtain benzyl (R)-methyl(2-(5-(1-((4-methyl-7-morpholinopyrido[3,4-d]pyridazin-1-yl)amino)ethyl)selenophene-3-yl)benzyl)carbamate (70.0 mg, 106 μmol, 24.1% yield) as a yellow solid.

[0190] MS (ESI) m / z 657.2 [M+H] +

[0191] Step (3) Preparation of (R)-4-methyl-N-(1-(4-(2-((methylamino)methyl)phenyl)selenophen-2-yl)ethyl)-7-morpholinopyrido[3,4-d]pyridazin-1-amine

[0192]

[0193] Benzyl (R)-methyl(2-(5-(1-((4-methyl-7-morpholinopyrido[3,4-d]pyridazin-1-yl)amino)ethyl)selenophene-3-yl)benzyl)carbamate (50.0 mg, 76.3 μmol, 1.00 eq) was dissolved in ethanol (2.00 mL) / water (0.200 mL), and sodium hydroxide (15.7 mg, 381.3 μmol, 45.7 μL, 5.00 eq) was added. The mixture was stirred at 70°C for 24 h and then distilled under reduced pressure to obtain the residue. After purification of the entire product by Prep-HPLC (column: Phenomenex luna C18 150×25 mm×10 μm; mobile phase: [water(formic acid)-acetonitrile; gradient: 8%–38% B for 9 min), lyophilization was performed to obtain (R)-4-methyl-N-(1-(4-(2-((methylamino)methyl)phenyl)selenophen-2-yl)ethyl)-7-morpholinopyrido[3,4-d]pyridazin-1-amine (10.32 mg, 19.35 μmol, 25.38% yield, 97.8% purity, formate) as a yellow solid.

[0194] 1 H NMR (400 MHz, DMSO-d6)δ= 9.04 (s, 1H), 8.23 ​​(s, 1H), 7.94 (s, 1H), 7.54 (br d,J= 6.4 Hz, 2H), 7.41 - 7.29 (m, 5H), 5.82 (br d,J= 6.0 Hz, 1H), 3.81 - 3.75 (m, 6H), 3.68 (br d,J= 4.8 Hz, 4H), 2.64 (s, 3H), 2.35 (s, 3H), 1.73 (d,J= 6.8 Hz, 3H).

[0195]

[0196] [Example 5] Preparation of (R)-N-(1-(4-(2-((dimethylamino)methyl)phenyl)selenophene-2-yl)ethyl)-6,7-dimethoxy-2-methylquinazolin-4-amine

[0197] It was manufactured in a similar manner to the manufacturing method of Example 1 according to the reaction formula below.

[0198]

[0199] 1 H NMR (400 MHz, DMSO-d6)δ= 8.16 (d,J= 8.0 Hz, 1H), 8.01 (d,J= 1.2 Hz, 1H), 7.64 (s, 1H), 7.49 (t,J= 1.2 Hz, 1H), 7.43 - 7.37 (m, 1H), 7.33 - 7.27 (m, 3H), 7.05 (s, 1H), 5.95 - 5.85 (m, 1H), 3.87 (d,J= 2.0 Hz, 6H), 3.28 (d,J= 5.2 Hz, 2H), 2.45 (s, 3H), 2.10 (s, 6H), 1.73 (d,J= 6.8 Hz, 3H).

[0200]

[0201] [Example 6] Preparation of (R)-6,7-dimethoxy-2-methyl-N-(1-(4-(2-(pyrrolidin-1-ylmethyl)phenyl)selenophen-2-yl)ethyl)quinazolin-4-amine

[0202] It was manufactured in a similar manner to the manufacturing method of Example 1 according to the reaction formula below.

[0203]

[0204] 1H NMR (400 MHz, DMSO-d6)δ= 14.69 - 14.11 (m, 1H), 9.96 - 9.77 (m, 1H), 9.87 (br d,J= 1.6 Hz, 1H), 8.06 (d,J= 1.2 Hz, 1H), 7.94 (s, 1H), 7.67 (dd,J= 3.6, 5.2 Hz, 1H), 7.53 - 7.45 (m, 2H), 7.43 (s, 1H), 7.40 - 7.32 (m, 1H), 7.21 (s, 1H), 6.07 (quin,J= 6.8 Hz, 1H), 4.37 (br s, 2H), 3.97 (s, 3H), 3.93 (s, 3H), 3.34 (br d,J= 4.8 Hz, 2H), 2.87 (br s, 2H), 2.65 (s, 3H), 1.80 (d,J= 7.2 Hz, 3H), 1.78 (br d,J= 3.6 Hz, 4H).

[0205]

[0206] [Example 7] Preparation of (R)-6,7-dimethoxy-2-methyl-N-(1-(4-(2-(((methyl-d3)amino)methyl)phenyl)selenophen-2-yl)ethyl)quinazolin-4-amine

[0207] It was manufactured in a similar manner to the manufacturing method of Example 1 according to the reaction formula below.

[0208] [Correction pursuant to Rule 91, 04.09.2024] 1 H NMR (400 MHz, DMSO-d6)δ= 8.21 (s, 2H), 8.17 (br d,J= 8.0 Hz, 1H), 7.96 (s, 1H), 7.64 (s, 1H), 7.52 (br d,J= 7.2 Hz, 1H), 7.41 (s, 1H), 7.38 - 7.29 (m, 3H), 7.05 (s, 1H), 5.90 (quin,J= 7.2 Hz, 1H), 3.87 (s, 6H), 3.76 (br s, 2H), 2.45 (s, 3H), 1.74 (d,J= 6.8 Hz, 3H).

[0209] [Example 8] Preparation of N-((R)-1-(4-(2-((dimethylamino)methyl)phenyl)selenophene-2-yl)ethyl)-7-methoxy-2-methyl-6-(((S)-tetrahydrofuran-3-yl)oxy)quinazolin-4-amine

[0210] It was manufactured in a similar manner to the manufacturing method of Example 2 according to the reaction formula below.

[0211] [Correction pursuant to Rule 91, 04.09.2024]

[0212] 1 H NMR (400 MHz, DMSO-d6)δ= 8.58 (d,J= 8.0 Hz, 1H), 8.46 (s, 1H), 8.07 (s, 1H), 7.93 (s, 1H), 7.85 (br d,J= 3.6 Hz, 1H), 7.76 - 7.73 (m, 3H), 7.52 (s, 1H), 6.39 - 6.34 (m, 1H), 5.58 - 5.55 (m, 1H), 4.42 - 4.39 (m, 1H), 4.32 (s, 3H), 4.31 - 4.23 (m, 3H), 3.73 (br d,J= 4.4 Hz, 2H), 2.90 (s, 3H), 2.74 - 2.71 (m, 1H), 2.55 (s, 6H), 2.46 (br s, 1H), 2.18 (d,J= 6.8 Hz, 3H).

[0213]

[0214] [Example 9] Preparation of 7-methoxy-2-methyl-N-((R)-1-(4-(2-(pyrrolidin-1-ylmethyl)phenyl)selenophen-2-yl)ethyl)-6-(((S)-tetrahydrofuran-3-yl)oxy)quinazolin-4-amine

[0215] It was manufactured in a similar manner to the manufacturing method of Example 2 according to the reaction formula below.

[0216]

[0217] 1H NMR (400 MHz, DMSO-d6)δ= 8.17 - 8.11 (m, 1H), 8.02 (s, 1H), 7.62 (s, 1H), 7.55 (s, 1H), 7.43 - 7.40 (m, 1H), 7.32 - 7.27 (m, 3H), 7.08 (s, 1H), 5.92 (br t,J= 7.2 Hz, 1H), 5.13 (br d,J= 4.4 Hz, 1H), 3.96 (dd,J= 4.8, 10.0 Hz, 1H), 3.88 (s, 3H), 3.86 - 3.74 (m, 3H), 3.52 - 3.44 (m, 2H), 2.45 (s, 3H), 2.39 (br s, 4H), 2.31 - 2.24 (m, 1H), 2.01 (br dd,J= 6.4, 12.8 Hz, 1H), 1.73 (d,J= 6.8 Hz, 3H), 1.63 (br s, 4H).

[0218]

[0219] [Experimental Example 1] Biochemical Analysis of SOS1 and KRAS G12C Interaction Inhibition Activity

[0220] In this experimental example, the efficacy of the compound according to the present invention to inhibit the binding of KRAS and SOS1 was biochemically measured. A KRAS G12C working solution was prepared by adding biotin-linked KRAS G12C protein and europium-linked streptavidin to an assay buffer (20 mM HEPES, 50 mM NaCl, 200 μg / mL fetal bovine serum (BSA), 0.01% Pluronic F127). Then, SOS1, in which a tag consisting of six histidine residues is covalently linked, and anti-histidine tag antibody D2 were added to the assay buffer to prepare an SOS1 working solution. After these two working solutions were added to the wells of a well plate, solutions diluted according to the experimental concentration of the test compound from the 10 mM dimethyl sulfoxide (DMSO) stock solution of the example compound or positive control compound were added to the wells in 100 nL aliquots using an Echo automatic dispenser (Beckman Coulter, USA), or a DMSO blank sample was added to the wells. After incubation for 60 minutes at room temperature, the fluorescence signal intensity was measured at two emission wavelengths using a PerkinElmer Envision instrument (excitation wavelength: 320 nm, emission wavelength: 665 / 615 nm). As positive control compounds, BAY-293 [(S)-6,7-dimethoxy-2-methyl-N-[1-[4-[2-[(methylamino)methyl]phenyl]thiophen-2-yl]ethyl]quinazolin-4-amine] or MRTX0902 [(R)-2-methyl-3-(1-((4-methyl-7-morpholinopyrido[3,4-d]pyridazin-1-yl)amino)ethyl)benzonitrile], a substance known as an SOS1 inhibitor, were used. Fluorescence measurement value y when the test compound concentration is x, and the minimum, maximum, and half-inhibitory concentrations (IC) of the fluorescence measurement and the binding activity were calculated. 50 ) determined from the concentration curve of inhibitory activity using the following relationship between 50 The values ​​are shown in Table 1 below.

[0221]

[0222]

[0223] [Experimental Example 2] Biochemical analysis of the inhibitory activity of SOS1 and KRAS G12D, G12V, G13D, or wild-type proteins, and SOS2 and KRAS G12C interactions.

[0224] In this experimental example, the interaction inhibition activity of KRAS G12C and SOS2 was biochemically measured to confirm the efficacy of the compound according to the present invention in inhibiting the binding of SOS1 to various types of mutant and wild-type KRAS and the selectivity for SOS2 enzyme. SOS1 or SOS2 protein covalently linked to a tag consisting of six histidine residues was added to the assay buffer (10 mM HEPES, 150 mM NaCl, 0.05% fetal bovine serum (BSA), 10 mM EDTA, 0.0005% Triton X100, 1 mM DTT). A solution diluted according to the experimental concentration of the test compound from a 10 mM dimethyl sulfoxide (DMSO) stock solution of the example compound or positive control compound was added to the well using an Echo automatic dispenser (Beckman Coulter, USA), or a DMSO blank sample was added to the well. After incubation at room temperature for 15 minutes, a working solution containing a mixture of biotin-linked KRAS, XL665-linked streptavidin (Cisbio 610SAXLB), and a detection solution containing terbium (Tb)-labeled anti-histidine tag antibody (Cisbio 61HI2TLA) was added to the assay buffer to prepare a working solution, which was added to the assay wells. Then, after incubation for 45–60 minutes at room temperature, the fluorescence signal intensity was measured at two emission wavelengths using a PHERAstar instrument from BMG Labtech (excitation wavelength: 337 nm, emission wavelength: 665 / 620 nm). BAY-293 or MRTX0902, which are known SOS1 inhibitors, were used as positive control compounds. The fluorescence measurement value y when the test compound concentration is x, and the minimum, maximum, and half-inhibitory concentration (IC) of the binding activity were calculated. 50 ) determined from the concentration curve of inhibitory activity using the following relationship between 50 The values ​​are shown in Table 2 below.

[0225]

[0226]

[0227]

[0228] [Experimental Example 3] Cell Growth Inhibition Efficacy Experiment

[0229] In this experimental example, the cell growth inhibitory effect of compounds according to the present invention and positive control compounds in tumor cell lines was examined.

[0230] NCI-H358 (US ATCC catalog number CRL-5807), a cancer cell line harboring KRAS G12C mutation, was cultured in a cell culture medium containing RPMI1640 (US Gibco catalog number 22400-089) and 10% fetal bovine serum (US Excell Bio catalog number FSP500), and 1,000 cells were seeded per well in a 96-well 3D assay plate and cultured for 1 day at 37°C and 5% CO2. From a 10-fold concentrated DMSO stock solution of the compound to be tested, a diluted amount of the test compound or DMSO blank was added to each well so that the final DMSO concentration was 0.25% in the culture medium, and this was cultured in a cell culture incubator for 12 days. 100 μL of the cell culture medium containing the compound was replaced every 3 days. Luminescence analysis was performed using the 3D CellTiter-Glo Luminescent Cell Viability Assay Kit (Promega, USA) (Promega catalog number G9681). Luminescence was measured using a PerkinElmer 2014 EnVision plate reader. The percent cell growth inhibition rate was calculated from the measured relative luminescence unit (RLU) as follows.

[0231]

[0232] From this cell growth inhibition rate, the half maximal inhibitory concentration (IC), which is the concentration at which each compound inhibits cell growth by 50%, was calculated using the GraphPad Prism program. 50 ) were calculated and summarized in Table 3 below.

[0233]

[0234] [Experimental Example 4] Experimental Effect of Combination with Targeted Anticancer Agents - Cell Viability

[0235] In this experimental example, we examined whether the compound according to the present invention has a synergistic effect on cancer cell death when used in combination with a target anticancer agent related to another RAS signal transduction pathway.

[0236] NCI-H358 cell line (ATCC catalog no. CRL-5807, USA) or EGFR exon 19 deletion cancer cell line PC-9 (RIKEN, Japan catalog no. RCB4455) were seeded in a well plate (500 cells per well for H358 and 50 cells per well for PC-9) in RPMI1640 (catalog no. FSP500, USA Excell Bio) containing 10% fetal bovine serum (catalog no. FSP500, USA). The cells were then cultured for one day at 37°C under 5% CO2 conditions. Subsequently, the target anticancer agent and the compound of the present invention were added in an amount of 150 μL according to the final experimental concentration so that the DMSO concentration in the culture medium was 0.25%. The targeted anticancer drugs used were sotorasib (Selleck, USA, catalog number S8830), a Ras inhibitor, trametinib (MedChemExpress, USA, catalog number HY10999), a MEK inhibitor, and osimertinib (MedChemExpress, USA, catalog number HY15772), an EGFR inhibitor. Cell survival was observed at six concentrations (10, 1, 0.1, 0.01, 0.001, and 0.0001 μM) for each of these compounds. Cell lines with added compounds were cultured in a cell culture medium for 7 days, and the cell culture medium containing the compounds was replaced on the 4th day of culture. Cell survival analysis was performed by luminescence analysis using the Promega CellTiter-Glo Luminescent Cell Viability Assay Kit (Promega, catalog number G7573), and luminescence was measured using a PerkinElmer 2014 EnVision plate reader. The inhibition rate was calculated from the measured relative luminescence value in the same manner as in Experimental Example 3, and the combination index (CI) between the compound of the present invention and the target anticancer agent was calculated using the following equation.

[0237]

[0238] In the above equation, CI(x) represents the concentration of the target anticancer agent and the compound of the present invention, respectively. and is the combination index when the cell growth inhibition rate measured in a combined experiment is x, is the concentration of the targeted anticancer drug required to achieve the same growth inhibition rate x when the targeted anticancer drug is used alone, is the concentration of the compound required to achieve the same growth inhibition rate x when the compound of the present invention is used alone.

[0239] In a combination test with a KRAS signal transduction blocking target anticancer drug targeting H358 cells, shown in Fig. 1, a strong synergistic effect was observed in all quantum concentration combinations between the Example 1 compound in the range of 0.0001 to 0.1 μM and the Example 1 compound in the range of 0.001 to 1 μM, and sotorasib, and cell growth was inhibited. On the other hand, trametinib showed a strong synergistic effect in all quantum concentration combinations between the Example 1 compound in the range of 0.0001 to 0.1 μM and the trametinib in the range of 0.001 to 0.1 μM, and cell growth was inhibited. In a combination test with an EGFR signal transduction blocking target anticancer drug targeting PC-9 cells, shown in Fig. 2, a strong synergistic effect was observed in all quantum combinations between the 0.01 to 0.1 μM range of osimertinib and the Example 1 compound in the range of 0.0001 to 10 μM.

[0240] From these results, it was expected that the compound of the present invention could block the binding of SOS1 and RAS, and would have a synergistic anticancer effect when used in combination with KRAS signaling pathway or EGFR signaling pathway inhibition therapy.

[0241] [Experimental Example 5] Experiment on the effect of generating reactive oxygen species

[0242] In this experimental example, it was examined that the compound according to the present invention has the effect of increasing active oxygen in tumor cells.

[0243] MIA-PaCa-2 (Korea Cell Line Bank KCLB No. 21420), a cancer cell line harboring KRAS G12C mutation, was cultured in a cell culture medium containing RPMI1640 (Gibco, USA, catalog no. 11875-119), 10% fetal bovine serum (HyClone, USA, catalog no. SH30919.03), and 1× diluted HyClone Antibiotic Antimycotic solution (HyClone, USA, catalog no. SV30079.01), and then seeded in a 6-well plate at a density of 3×10 per well. 5 After seeding the cells, they were cultured for one day at 37°C and 5% CO2. The test compounds or DMSO blanks were added to each well so that the final concentration of the compound to be tested was 10 μM and the final DMSO concentration was 0.2% in the culture medium, and they were cultured in a cell incubator for 24 hours. After washing once with phosphate-buffered saline (PBS, Gibco, USA, catalog number 10010-049), the wells were treated with 20 μM 2',7'-dichlorodihydrofluorescein diacetate (Sigma-Aldrich, USA, catalog number D6883) for 30 minutes. The cells were then collected in a microtube and analyzed using a BD LSRFortessa, a flow cytometer from BD Biosciences, USA. TMFluorescence was analyzed using an X-20 Cell Analyzer (excitation wavelength: 488 nm, emission wavelength: 535 nm). The fluorescence intensity due to the increase in reactive oxygen species was measured for 10,000 cells. Among the cells treated with 500 μM hydrogen peroxide, which was a positive control, the section in which the fluorescence intensity increased compared to the blank-treated cells was found, and the average fluorescence intensity of the cells within this section was calculated. Then, the average of the increased fluorescence intensity measurements compared to the blank within the same section in the test compound-treated cells was converted into a percentage of the average of the positive control group.

[0244] In the graph of the rate of reactive oxygen species production shown in Fig. 3, the compound of Example 1 increased strongly compared to the blank sample, and the control drug MRTX0902 did not increase the rate of reactive oxygen species production compared to the blank sample.

[0245] From these results, it was expected that the compound of the present invention increases intracellular reactive oxygen species, which enhances the cell growth inhibitory effect.

[0246] Although the present invention has been described through limited embodiments as described above, the technical idea and scope of the present invention are not limited to these embodiments, and the invention of the patent claims described below, as well as various modifications or variations of the invention described in the patent claims that are obvious to a person having ordinary skill in the art to which the present invention pertains, are also included in the scope of the present invention within the equivalent scope of the invention described in the patent claims.

Claims

1. A compound of chemical formula 1 or a totoisomer thereof and a pharmaceutically acceptable salt thereof: [Chemical Formula 1] In the above chemical formula 1, R 1 and R 2 are H and C respectively 1~4 Alkyl and deuterium substituted C 1~4 A functional group independently selected from the group consisting of alkyl, or R 1 and R 2 is R 1 and R 2 Together with the nitrogen atom to which it is bonded, it forms a 4-6 membered heterocycloalkyl ring, where R 1 and R 2 Except when both are H at the same time, Is And at this time Q 1 and Q 2 are each independently C 1~4 A functional group selected from alkyl and 4-8 membered single ring heterocyclyl, and Q 3 is a 4-8 membered single ring heterocyclyl, The sign is It refers to the connection at that position to the rest of the compound.

2. A compound according to claim 1, characterized in that the 4-6 membered heterocycloalkyl ring is azetidinyl or pyrrolidinyl.

3. In paragraph 1, Q 1 and Q 2 are all methyl, or Q 1 and Q 2 A compound characterized in that one of the groups is methyl and the other is tetrahydrofuranyl.

4. Any one compound selected from the group consisting of the following compounds, a stereoisomer thereof, a tautomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof: (R)-6,7-dimethoxy-2-methyl-N-(1-(4-(2-((methylamino)methyl)phenyl)selenophen-2-yl)ethyl)quinazolin-4-amine; 7-Methoxy-2-methyl-N-((R)-1-(4-(2-((methylamino)methyl)phenyl)selenophen-2-yl)ethyl)-6-(((S)-tetrahydrofuran-3-yl)oxy)quinazolin-4-amine; (R)-(2-chloro-4-((1-(4-(2-((methylamino)methyl)phenyl)selenophene-2-yl)ethyl)amino)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(4-methoxytetrahydro-2H-pyran-4-yl)methanone; (R)-4-methyl-N-(1-(4-(2-((methylamino)methyl)phenyl)selenophen-2-yl)ethyl)-7-morpholinopyrido[3,4-d]pyridazin-1-amine; (R)-N-(1-(4-(2-((dimethylamino)methyl)phenyl)selenophene-2-yl)ethyl)-6,7-dimethoxy-2-methylquinazolin-4-amine; (R)-6,7-dimethoxy-2-methyl-N-(1-(4-(2-(pyrrolidin-1-ylmethyl)phenyl)selenophen-2-yl)ethyl)quinazolin-4-amine; (R)-6,7-dimethoxy-2-methyl-N-(1-(4-(2-(((methyl-d3)amino)methyl)phenyl)selenophen-2-yl)ethyl)quinazolin-4-amine; N-((R)-1-(4-(2-((dimethylamino)methyl)phenyl)selenophene-2-yl)ethyl)-7-methoxy-2-methyl-6-(((S)-tetrahydrofuran-3-yl)oxy)quinazolin-4-amine and 7-Methoxy-2-methyl-N-((R)-1-(4-(2-(pyrrolidin-1-ylmethyl)phenyl)selenophen-2-yl)ethyl)-6-(((S)-tetrahydrofuran-3-yl)oxy)quinazolin-4-amine.

5. A pharmaceutical composition for the treatment of cancer comprising a therapeutically effective amount of a compound according to claim 1 and a pharmaceutically acceptable excipient.

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