1,5-Naphthyridine derivatives as KRAS oncoprotein inhibitors

Abstract

The present invention is directed to inhibitors of Kirsten rat sarcoma virus (KRAS) oncoprotein, and more particularly to inhibitors of certain formula I: The present invention is directed to 1,5-naphthyridine derivatives of formula I, as well as compositions comprising the compounds of formula I and methods of using the compounds of formula I for the treatment or prevention of diseases, disorders, or medical conditions mediated by KRAS, particularly KRAS G12C or KRAS G12D oncoprotein.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. 63 / 420,831, filed October 31, 2022, and U.S. Patent Application No. 63 / 510,236, filed June 26, 2023, both of which are incorporated herein by reference in their entireties. [Background technology]

[0002] Background of the Invention Technical field of the invention The present invention is directed to inhibitors of Kirsten rat sarcoma virus (KRAS) oncoprotein, and more particularly to certain 1,5-naphthyridine derivatives, compositions, and methods for the treatment or prevention of diseases, disorders, or medical conditions mediated by KRAS, particularly KRAS G12C and KRAS G12D oncoproteins, including various cancers.

[0003] Brief description of the related art Ras is a superfamily of small guanosine triphosphate (GTP)-binding proteins consisting of various isoforms. Ras genes can mutate into oncogenes associated with numerous cancers, including lung, pancreatic, and colon cancer. Ras is one of the most frequently mutated oncogenes. The Ras isoform KRAS (Kirsten rat sarcoma virus) is one of the most frequently mutated Ras genes, accounting for approximately 86% of all known mutations. KRAS functions as an on / off switch in cell signaling. KRAS proteins are GTPases that act between an inactive (GDP-bound) and an active (GTP-bound) state to regulate various functions, including cell proliferation. However, mutated KRAS proteins can lead to uncontrolled cell proliferation and cancer. The KRAS-4B proteoform is the predominant isoform in colon cancer (30-40%), lung cancer (15-20%), and pancreatic cancer (90%) (Liu, P. et al., Acta Pharmaceutica Sinica B 2019, 9(5), 871-879). Therefore, inhibitors of mutated GTP-binding KRAS proteins represent potential therapeutic agents for the treatment of various cancers.

[0004] Previous attempts to design KRAS oncoprotein inhibitors have met with little success, largely due to the high affinity of the KRAS oncoprotein for GTP. However, more recent approaches targeting KRAS G12C have shown more promise. This mutation is present in approximately 50% of lung cancers and approximately 10–20% of all KRAS G12 mutations. The cysteine ​​residue in this mutation is positioned within the active site so that the sulfhydryl functional group can form a covalent bond with an appropriately functionalized binding ligand (Liu, Acta Pharmaceutica Sinica B 2019). This approach has identified irreversible covalent inhibitors of KRAS G12C, which are currently in clinical trials. KRAS G12D mutations are found in approximately 4% of all non-small cell lung cancers, 13% of all colorectal cancers, 25% of pancreatic ductal adenocarcinomas, and 1.7% of small cell lung cancers (Cerami, E. and Sawyers, CL Cancer Discovery 2017, 7(8), 818-831). Given the prominent roles of both KRAS G12C and KRAS G12D as drivers of many malignancies, new KRAS G12C and KRAS G12D inhibitors with improved selectivity, safety, and efficacy profiles are needed. Summary of the Invention

[0005] In one embodiment, the present invention provides a compound of formula I: [ka] [During the ceremony, A is hydrogen, halogen, hydroxy, -C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 Alkynyl, -C 0-3 Alkyl (C 3-6 cycloalkyl), -C 1-6 Alkyl (halo), -C 1-6 Alkyl (OH), -O(C 1-4 alkyl), -C 1-3 Alkyl (C 1-4Alkoxy), -CN, -CO2R 4 , -CO2N(R 4 )2, -NO2, -N(R 4 )2, -P(O)(R 5 )2, -SR 4 , -S(O)R 4 , -SO2R 4 or aryl or heteroaryl optionally substituted with one or more 5- to 6-membered heterocyclic rings; Y and G may be the same or different and are hydrogen, halogen, C 1-4 Alkyl, C 1-4 Perdeuteroalkyl, -(C 0-2 alkyl)alkenyl, -(C 0-2 alkyl)alkynyl, -(C 0-2 alkyl)cycloalkyl, -C 1-4 Haloalkyl, -O(C 1-4 alkyl), -S(C 1-4 alkyl), -(C 0-2 alkyl) cyano, -O(C 1-4 haloalkyl) or -S(C 1-4 haloalkyl); L is a bond, O, S, or NR 4 and; m is 0 to 2; n is 0 to 2; Z is C(R 4 )2, or C 3-7 a cyclic compound selected from cycloalkyl, a saturated or partially unsaturated 4- to 7-membered nitrogen-containing ring, and a saturated or partially unsaturated 7- to 10-membered nitrogen-containing bridged bicyclic ring; R 1 is hydrogen, hydroxy, halogen, -C 1-3 Alkyl, -C 1-3 Alkyl (OH), -C 1-3 Alkyl (halo), -C 1-3 Alkyl (C 1-3 alkoxy), -C 1-3 Alkyl (CN) or -C 1-3 Alkyl (P(O)R 5 2) Selected from; R2 is selected from hydrogen, —C(O)CH═CH, —C(O)CF═CH or —C(O)CCl═CH, with the proviso that R 2 is hydrogen, m is 1 or 2; R 3 is hydrogen, halogen, hydroxy, -C 1-4 Alkyl, -C 2-4 Alkenyl, -C 2-4 Alkynyl, -C 0-3 Alkyl (C 3-6 cycloalkyl), -C 1-4 Alkyl (halo), -C 1-4 Alkyl (OH), -O(C 1-4 alkyl), -C 1-3 Alkyl (C 1-3 Alkoxy), -CN, -CO2R 4 , -CO2N(R 4 )2, -NO2, -N(R 4 )2, -PO(R 5 )2, -SR 4 , -S(O)R 4 , -SO2R 4 or -(C 0-3 alkyl)R 6 Selected from; R 4 is hydrogen, C 1-4 selected from alkyl, aryl, or heteroaryl; R 5 is hydrogen, hydroxy, C 1-4 Alkyl, aryl, heteroaryl, C 1-4 selected from alkoxy, aryloxy, or heteroaryloxy; R 6 is N(R 4 )2, or a 4- to 7-membered saturated or unsaturated heterocyclic ring containing one or more heteroatoms selected from the group N, O, and S. or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0006] In another aspect, the present invention is directed to a pharmaceutical composition comprising a compound of Formula I, or a salt, solvate, or prodrug thereof, in association with a pharmaceutically acceptable carrier.

[0007] In another aspect, the present invention is directed to a method of treating a disease, disorder, or medical condition in a patient, said method comprising providing a therapeutic agent to a patient in need of treatment, wherein said therapeutic agent is a compound of formula I or a salt, solvate, or prodrug thereof. DETAILED DESCRIPTION OF THE INVENTION

[0008] Detailed Description of the Invention term Compounds are described using standard nomenclature. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0009] The use of the terms "a," "an," or "the" does not denote a limitation of quantity, but rather denotes the presence of at least one of the referenced item. The term "or" means "and / or." The terms "including," "containing," and "having" are intended to be open-ended (i.e., meaning "including, but not limited to").

[0010] Recitation of ranges of values, unless otherwise stated herein, is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, and each separate value is incorporated herein as if it were individually set forth herein. All range endpoints are included within the ranges and are independently combinable.

[0011] All methods described herein can be performed in any suitable order unless otherwise indicated herein or clearly contradicted by context. The use of any and all examples or exemplary language (e.g., "such as") is intended merely to better illustrate the present invention and does not limit the scope of the present invention unless otherwise claimed. No term in this specification should be construed as indicating any non-claimed element as essential to the practice of the invention used herein. Unless otherwise defined, technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art of this disclosure.

[0012] Furthermore, the disclosure encompasses all variations, combinations, and permutations in which one or more limitations, elements, clauses, and descriptive terms of one or more enumerated claims are introduced into another claim. For example, any claim that is dependent on another claim can be modified to include one or more limitations found in another claim that is dependent on the same base claim. Where elements are presented as lists, e.g., in Markush group format, each subgroup of elements is also disclosed, and any element can be removed from the group.

[0013] All compounds are understood to include all possible isotopes of the atoms present in the compound. Isotopes include atoms having the same atomic number but different mass numbers, and include heavy isotopes and radioisotopes. Common examples include isotopes of hydrogen include tritium and deuterium, and isotopes of carbon include 11 C. 13 C, and 14 These include, but are not limited to, C. Thus, the compounds disclosed herein may contain heavy or radioisotopes in the structure of the compound or as substituents attached thereto. Examples of useful heavy or radioisotopes include: 18 F, 15 N, 18 O. 76 Br, 125 I and 131 Contains I.

[0014] All formulas disclosed herein include all salts of that formula.

[0015] The open-ended term "comprising" includes the intermediate and closed terms "consisting essentially of" and "consisting of."

[0016] The term "substituted" means that any one or more hydrogens on the specified atom or group are replaced with a selection from the indicated group, provided that the normal valence of the specified atom is not exceeded. Combinations of substituents and / or variables are permissible only if such combinations result in stable compounds or useful synthetic intermediates. A stable compound or stable structure is intended to imply a compound that has sufficient robustness to survive isolation from a reaction mixture, and subsequent formulation into an effective therapeutic agent.

[0017] A dash ("-") that is not between two letters or symbols is used to indicate a point of attachment for a substituent.

[0018] "Alkyl" includes both branched and straight-chain saturated aliphatic hydrocarbon groups having a specified number of carbon atoms, generally from 1 to about 8 carbon atoms. 1-6 The terms alkyl, C1-C6 alkyl and C1-C6 alkyl all refer to alkyl groups having 1, 2, 3, 4, 5, or 6 carbon atoms. Other embodiments include alkyl groups having 1 to 8 carbon atoms, 1 to 4 carbon atoms, or 1 or 2 carbon atoms, such as C 1-8 Alkyl, C 1-4 Alkyl, and C 1-2 As used herein, C 0-n When alkyl is used with other groups, e.g., -C 0-4In alkyl(phenyl), the indicated group, in this case phenyl, is either directly attached by a single covalent bond (CO alkyl) or by an alkyl chain having the specified number of carbon atoms, in this case 1, 2, 3, or 4 carbon atoms. Alkyl is represented by -OC 0-4 Alkyl (C 3-7 As in cycloalkyl, it may also be attached through other groups such as a heteroatom. Examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, 3-methylbutyl, t-butyl, n-pentyl, and sec-pentyl.

[0019] "Alkoxy" refers to an alkyl group as defined above having the indicated number of carbon atoms covalently linked to a substituent through an oxygen bridge (-O-). Examples of alkoxy include, but are not limited to, methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, 2-butoxy, t-butoxy, n-pentoxy, 2-pentoxy, 3-pentoxy, isopentoxy, neopentoxy, n-hexoxy, 2-hexoxy, 3-hexoxy, and 3-methylpentoxy. Similarly, an "alkylthio" or "thioalkyl" group refers to an alkyl group as defined above having the indicated number of carbon atoms covalently linked to a substituent through a sulfur bridge (-S-). Similarly, "alkenyloxy," "alkynyloxy," and "cycloalkyloxy" refer to alkenyl, alkynyl, and cycloalkyl groups, respectively, which are covalently linked to a substituent through an oxygen bridge (-O-).

[0020] "Halo" or "halogen" means fluoro, chloro, bromo, or iodo, and is defined herein to include all isotopes thereof, including heavy and radioactive isotopes. Examples of useful halo isotopes include: 18 F, 76 Br, and 131 I. Additional isotopes will be readily apparent to those skilled in the art.

[0021] "Haloalkyl" refers to both branched and straight-chain alkyl groups having the specified number of carbon atoms and substituted with one or more halogen atoms, generally up to the maximum allowable number of halogen atoms. Examples of haloalkyl include, but are not limited to, trifluoromethyl, difluoromethyl, 2-fluoroethyl, and pentafluoroethyl.

[0022] "Haloalkoxy" is a haloalkyl group as described above attached through an oxygen bridge (oxygen of an alcohol radical).

[0023] "Peptide" means a molecule that is a chain of amino acids linked through amide bonds (also called peptide bonds).

[0024] "Pharmaceutical composition" means a composition comprising at least one active agent, such as a compound or salt of Formula I, and at least one other substance, such as a carrier. The pharmaceutical composition meets U.S. FDA Good Manufacturing Practice (GMP) standards for human or non-human pharmaceuticals.

[0025] "Carrier" refers to a diluent, excipient, or vehicle with which an active compound is administered. "Pharmaceutically acceptable carrier" refers to a substance, e.g., an excipient, diluent, or vehicle, useful in preparing pharmaceutical compositions that are generally safe, non-toxic, and not biologically or otherwise undesirable, and includes carriers acceptable for veterinary and human pharmaceutical use. "Pharmaceutically acceptable carrier" includes both one such carrier and more than one such carrier.

[0026] "Patient" means a human or non-human animal in need of medical treatment. Medical treatment can include treatment of an existing condition, such as a disease or disorder, or diagnostic treatment. In some embodiments, the patient is a human patient.

[0027] "Providing" means giving, administering, selling, distributing, giving away (whether for profit or not), manufacturing, compounding, or distributing.

[0028] "Treatment" or "treating" means providing a patient with a sufficient amount of an active compound to measurably alleviate any disease symptoms, slow the progression of the disease, or cause regression of the disease. In certain embodiments, treatment of a disease can begin before the patient exhibits symptoms of the disease.

[0029] A "therapeutically effective amount" of a pharmaceutical composition means an amount that, when administered to a patient, is effective to provide a therapeutic benefit, such as amelioration of symptoms, reduction in progression of a disease, or regression of a disease.

[0030] "Therapeutic compound" means a compound that can be used in the diagnosis or treatment of disease. The compound can be a small molecule, peptide, protein, or other type of molecule.

[0031] A significant change is any detectable change that is statistically significant in a standard parametric test of statistical significance, such as Student's T-test, where p<0.05.

[0032] chemical description The compounds of the formulas disclosed herein may contain one or more asymmetric elements, such as a stereogenic center (e.g., an asymmetric carbon atom), a stereogenic axis, or a rotationally hindered rotamer (e.g., atropisomers), such that the compounds can exist in different stereoisomeric forms. These compounds may be, for example, racemic or optically active. For compounds with two or more asymmetric elements, these compounds may further be mixtures of diastereomers. For compounds with asymmetric centers, all optical isomers in pure form and mixtures thereof are encompassed. In these situations, single enantiomers, i.e., optically active forms, can be obtained by asymmetric synthesis, synthesis from optically pure precursors, or separation of the racemate. Separation of the racemate can also be achieved by conventional methods, such as crystallization in the presence of a resolving agent or chromatography, e.g., using a chiral HPLC column. All forms are contemplated herein, regardless of the method used to obtain them.

[0033] All forms of the compounds of the present invention (eg, solvates, optical isomers, enantiomeric forms, polymorphs, prodrugs, free base compounds and salts) may be used alone or in combination.

[0034] The term "chiral" refers to a molecule having the property of non-superimposability of its mirror image partner.

[0035] "Stereoisomers" are compounds which have identical chemical constitution, but differ with regard to the arrangement of the atoms or groups in space.

[0036] The term "solvate" refers to a chemical complex formed by the interaction of a solvent and a solute, such as a compound of the invention.

[0037] The term "prodrug" refers to a biologically inactive compound that can be metabolized in the body to produce a drug.

[0038] "Diastereomers" are stereoisomers that have two or more centers of chirality and whose molecules are not mirror images of one another. Diastereomers have different physical properties, e.g., melting points, boiling points, spectral properties, and reactivities. Mixtures of diastereomers can be separated by high-resolution analytical procedures such as electrophoresis, crystallization in the presence of a resolving agent, or chromatography, e.g., using a chiral HPLC column.

[0039] "Enantiomers" refer to two stereoisomers of a compound that are non-superimposable mirror images of one another. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, and may occur where there has been no stereoselection or stereospecificity in a chemical reaction or process.

[0040] The stereochemical definitions and conventions used herein are generally those of SP Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., Stereochemistry of Organic Compounds (1994) John Wiley & Sons, Inc., New York. Many organic compounds exist in optically active forms, i.e., they have the ability to rotate the plane of plane-polarized light. In describing optically active compounds, the prefixes D and L, or R and S, are used to indicate the absolute configuration of the molecule about its chiral center. The prefixes d and l, or (+) and (-) are used to indicate the sign of rotation of plane-polarized light by the compound, with (-) or l meaning the compound is levorotatory. Compounds with a (+) or d prefix are dextrorotatory.

[0041] A "racemic mixture" or "racemate" is an equimolar (or 50:50) mixture of two enantiomeric species, devoid of optical activity. A racemic mixture may occur where there has been no stereoselection or stereospecificity in a chemical reaction or process.

[0042] A "chelating group" or "chelator" is a ligand group that can form two or more separate coordinate bonds to a single central atom, usually a metal ion. The chelating groups disclosed herein are organic groups that have multiple N, O, or S heteroatoms and a structure in which the two or more heteroatoms can form bonds to the same metal ion.

[0043] "Salts" include derivatives of the disclosed compounds modified by converting the parent compound into its inorganic and organic salts, acid addition salts, or base addition salts. Salts of the compounds of the present invention can be synthesized from parent compounds containing a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid form of these compounds with a stoichiometric amount of an appropriate base (such as hydroxide, carbonate, or bicarbonate of Na, Ca, Mg, or K), or by reacting the free base form of these compounds with a stoichiometric amount of an appropriate acid. Such reactions are typically carried out in water or an organic solvent, or a mixture of both. Non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are generally used, where feasible. Salts of the compounds of the present invention also include solvates of the compounds of the present invention and solvates of salts of the compounds of the present invention. In certain embodiments, the compounds of the present invention are synthesized or isolated as trifluoroacetic acid (TFA) salts.

[0044] In some embodiments, the salt form of the above-mentioned compound of the present invention can include pharmaceutically acceptable salt.The example of pharmaceutically acceptable salt includes but is not limited to the non-toxic mineral acid salt or organic acid salt of basic residue such as amine; the alkali salt or organic acid salt of acidic residue such as carboxylic acid; etc.Pharmaceutically acceptable salt includes conventional salt and, for example, the quaternary ammonium salt of parent compound formed from non-toxic inorganic acid or organic acid. For example, conventional non-toxic acid salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, and nitric acid; and acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, mesylic acid, esylic acid, besylic acid, sulfanilic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, isethionic acid, and HOOC-(CH2) n These include salts prepared from organic acids such as —COOH, where n is 0 to 4. Further lists of suitable salts can be found, for example, in G. Steffen Paulekuhn, et al., Journal of Medicinal Chemistry 2007, 50, 6665 and Handbook of Pharmaceutically Acceptable Salts: Properties, Selection and Use, P. Heinrich Stahl and Camille G. Wermuth, Editors, Wiley-VCH, 2002.

[0045] In a preferred embodiment, compounds of Formula I are represented by structures 1a-1ah and 2a-2aw shown below, including pharmaceutically acceptable salts, solvates, or prodrugs thereof: [ka] [ka] [ka] [ka] [ka] [ka]

[0046] Particularly preferred compounds of the above are 1ae, 1af, 1ag, 1ah, 1b, 1d, 1i, 1q, 1s, 1x, 2z, 2al, 2an, 2ap and 2aq: [ka] is.

[0047] The compounds disclosed herein can be administered to a patient as neat or free-base chemicals, but are preferably administered as pharmaceutical compositions. Accordingly, the present invention encompasses pharmaceutical compositions comprising a compound, such as a compound of Formula I, or a salt of the compound (including a pharmaceutically acceptable salt), in combination with at least one pharmaceutically acceptable carrier. The pharmaceutical composition may comprise a compound of Formula I or a salt thereof as the sole active agent, but preferably also comprises at least one additional active agent. In certain embodiments, the pharmaceutical composition comprises about 0.1 mg to about 2000 mg, about 10 mg to about 1000 mg, about 100 mg to about 800 mg, or about 200 mg to about 600 mg of a compound of Formula I in a unit dosage form, and optionally about 0.1 mg to about 2000 mg, about 10 mg to about 1000 mg, about 100 mg to about 800 mg, or about 200 mg to about 600 mg of an additional active agent in the unit dosage form. Pharmaceutical compositions can also include a molar ratio of a compound, such as the compound of Formula I, to an additional active agent. For example, the pharmaceutical composition can include a molar ratio of about 0.5:1, about 1:1, about 2:1, about 3:1, or about 1.5:1 to about 4:1 of the additional active agent to the compound of Formula I. Particularly preferred forms of the compound of Formula I for use in pharmaceutical compositions include compounds 1ae-1ah, 1b, 1d, 1q, 2z, or 2al, or salts, solvates, or prodrugs thereof, together with a pharmaceutically acceptable carrier.

[0048] The compounds disclosed herein can be administered orally, topically, parenterally, by inhalation or spray, sublingually, transdermally, buccal administration, rectally, as eye drops, or by other means in dosage unit formulations containing conventional pharmaceutically acceptable carriers. Pharmaceutical compositions can be formulated into any pharmaceutically useful form, for example, aerosols, creams, gels, pills, capsules, tablets, syrups, transdermal patches, or eye drops. Some dosage forms, such as tablets and capsules, are subdivided into unit doses of suitable size containing an appropriate amount of active ingredient, for example, an amount effective to achieve the desired purpose.

[0049] Carriers include excipients and diluents and must be sufficiently pure and sufficiently low in toxicity to be suitable for administration to the patient being treated. Carriers may be inert or may have pharmaceutical benefits of their own. The amount of carrier used with the compound is sufficient to provide a practical amount of material for administration per unit dose of the compound.

[0050] Types of carriers include, but are not limited to, binders, buffers, colorants, diluents, disintegrants, emulsifiers, flavorings, glidants, lubricants, preservatives, stabilizers, surfactants, tableting agents, and wetting agents. Some carriers may be classified into multiple categories; for example, vegetable oils may be used as lubricants in some formulations and as diluents in other formulations. Exemplary pharmaceutically acceptable carriers include sugars, starches, cellulose, powdered tragacanth, malt, gelatin, talc, and vegetable oils. The pharmaceutical composition may contain any active agent that does not substantially inhibit the activity of the compound of the present invention.

[0051] The pharmaceutical compositions / combinations can be formulated for oral administration. These compositions contain 0.1 to 99% by weight (wt%) of the compound of formula I, and usually contain at least about 5 wt% of the compound of formula I. Some embodiments contain about 25 wt% to about 50 wt% or about 5 wt% to about 75 wt% of the compound of formula I.

[0052] Treatment method The compounds of Formula I and pharmaceutical compositions comprising said compounds are useful for the diagnosis or treatment of diseases, disorders or medical conditions mediated by KRAS, particularly KRAS mutants G12C and G12D, including various cancers such as glioma (glioblastoma), acute myelogenous leukemia, acute myeloid leukemia, myelodysplastic / myeloproliferative neoplasms, sarcoma, chronic myelomonocytic leukemia, non-Hodgkin's lymphoma, astrocytoma, melanoma, non-small cell lung cancer, cholangiocarcinoma, chondrosarcoma, colon cancer, or pancreatic cancer.

[0053] According to the present invention, the method for treating KRAS-mediated disease or condition comprises providing the patient who needs such treatment with a therapeutically effective amount of the compound of formula I. In some embodiments, the patient is a mammal, more particularly, a human.As will be understood by those skilled in the art, the present invention also encompasses the method for treating non-human patients, such as pets, for example, cats, dogs and livestock.

[0054] The therapeutically effective amount of pharmaceutical composition is preferably sufficient to alleviate or improve the symptoms of disease or condition.For example, in the case of KRAS-mediated disease, the therapeutically effective amount can be sufficient to alleviate or improve cancer.The therapeutically effective amount of compound or pharmaceutical composition described herein also provides sufficient concentration of the compound of Formula I when administered to patient.Sufficient concentration is preferably the concentration of compound in the patient's body that is required to prevent or combat disorder.Such amount can be confirmed experimentally, for example, by measuring the blood concentration of compound, or theoretically by calculating bioavailability.

[0055] According to the present invention, the treatment methods disclosed herein comprise providing a patient with a specific dosage of a compound of Formula I. Dosage levels of about 0.1 mg to about 140 mg of each compound per kg of body weight per day are useful for treating the above conditions (about 0.5 mg to about 7 g per patient per day). The amount of compound that can be combined with a carrier material to produce a single dosage form varies depending on the patient being treated and the particular mode of administration. Dosage unit forms generally contain about 1 mg to about 500 mg of each active compound. In certain embodiments, 25 mg to 500 mg, or 25 mg to 200 mg, of a compound of Formula I is provided to a patient daily. The frequency of administration can also vary depending on the compound used and the particular disease being treated. However, for the treatment of most KRAS-mediated diseases and disorders, a dosing regimen of four times per day or less can be used, and in certain embodiments, a once- or twice-daily dosing regimen is used.

[0056] However, it will be understood that the specific dose level for any particular patient will depend on a variety of factors, including the activity of the particular compound used, age, body weight, general health, sex, dietary habits, time of administration, route of administration and excretion rate, drug combination, and the severity of the particular disease being treated.

[0057] The compound of Formula I can be administered alone (i.e., the only therapeutic agent in the regimen) or in combination with another active agent to treat or prevent KRAS-mediated diseases and conditions, such as various cancers.One or more compounds of Formula I can be administered in concert with one or more other active agent regimens, such as anti-cancer cytotoxic agents.In some embodiments, the method for treating or diagnosing KRAS-mediated cancer in a mammal comprises administering to the mammal a therapeutically effective amount of a compound of Formula I, optionally in combination with one or more additional active ingredients.

[0058] As will be appreciated by those skilled in the art, the methods of treatment provided herein are also useful for treating mammals other than humans, including veterinary applications such as treating horses and livestock, e.g., cattle, sheep, cows, goats, pigs, etc., as well as companion animals such as dogs and cats.

[0059] For diagnostic or research applications, a wide variety of mammals are suitable subjects, including rodents (e.g., mice, rats, hamsters), rabbits, primates, and pigs, such as inbred pigs. Additionally, for in vitro applications, such as in vitro diagnostic and research applications, bodily fluids (e.g., blood, plasma, serum, interstitial fluid, saliva, feces, and urine) and cell and tissue samples from the subjects are suitable for use.

[0060] In certain embodiments, the present invention provides a method of treating a disease, disorder, or medical condition mediated by KRAS, particularly the KRAS mutant G12C, including various cancers, in a patient identified as in need of such treatment, the method comprising providing to the patient an effective amount of a compound of Formula I. The compounds of Formula I provided herein can be administered alone or in combination with one or more other active agents.

[0061] In another embodiment, the method of treating or diagnosing a KRAS-mediated disease or condition can further comprise administering to a patient in need of such treatment a compound of Formula I in combination with one or more additional compounds, wherein at least one of the additional compounds is an active agent. The one or more additional compounds can be selected from the group consisting of doxorubicin, paclitaxel, docetaxel, cisplatin, camptothecin, temozolomide, Avastin, Herceptin, Erbitux, EGFR inhibitors, osimertinib, rezivertinib, CDK4 / 6 inhibitors, abemaciclib, palbociclib, ribociclib, c-MET inhibitors, capmatinib, volitinib, and the like. ), ALK inhibitors, crizotinib, alectinib, ceritinib, brigatinib, entrectinib, lorlatinib, PD-1 antagonists, PD-L1 antagonists, ipilimumab, embrolizumab, and nivolumab, among other anti-cancer therapeutic compounds. [Example]

[0062] chemical synthesis The compounds of Formula 1 and / or pharmaceutically acceptable salts thereof described herein can be synthesized from commercially available starting materials by methods well known to those skilled in the art of organic synthetic chemistry. The following general synthetic schemes 1 and 2 show representative methods for preparing most of the exemplary compounds. In certain instances where the Suzuki cross-coupling reaction of arylboronic acids / esters with organic halides / pseudohalides (Beketskaya, IP et al., Coordin. Chem. Rev. 2019, 385, 137-173) is impractical or unsuccessful, the corresponding Stille cross-coupling reaction of organostannanes with organic halides / pseudohalides can be used as an alternative (Espinet, P. et al., ACS Catal. 2015, 5, 3040-3053). Many of the required intermediates can be prepared as described in WO2021041671. The starting materials, reactions, reagents, solvents, temperatures, catalysts, and ligands described are not limited to those depicted purely for illustrative purposes. Certain abbreviations and acronyms that may be used in Schemes 1 and 2 and the Examples, and that are well known to those of skill in the art, are set forth below for clarity.

[0063] The synthesis of compounds of the present invention is exemplified by the sequence of steps shown in Scheme 1. In Scheme 1, oxidation of 1,5-naphthyridine derivative 3 with mCPBA in a solvent such as CHCl generates N-oxide compound 4. Reaction of 4 with POCl at elevated temperatures affords the corresponding chloro derivative 5. Reaction of 5 with 6 affords compound 7. Treatment of 8 with a suitable base such as sodium hydride, Hunig's base, KCO, or a CsCO / DABCO mixture, followed by reaction with 7 in a polar aprotic solvent such as N-methyl-2-pyrrolidone at room temperature (RT) or elevated temperatures, affords compound 9. Suzuki-Miyaura coupling of 9 with a boronic ester such as 10 (or the corresponding boronic acid) under standard conditions in a solvent mixture such as 1,4-dioxane and water can be used to prepare 11. The Boc protecting group of 11 can be removed under acidic conditions such as anhydrous 4 M HCl in 1,4-dioxane or TFA in CH2Cl2 to give R 2 Acylation of 12 with acryloyl chloride 13 in a solvent such as methylene chloride containing a base such as triethylamine gives compounds of formula I (12) where R 2 The corresponding compound 14 of formula I, where is either —C(O)CH═CH, —C(O)CF═CH, or —C(O)CCl═CH, is produced. [ka]

[0064] Abbreviations and Acronyms The following abbreviations and acronyms may be used herein: anhyd.=anhydrous; aq. = aqueous; B2pin2=bis(pinacolato)diboron; Boc = tert-butoxycarbonyl; n-Bu3P = tri-n-butylphosphine; Compd=Compound; d=day; DCM = dichloromethane; DIEA = DIPEA = N,N-diisopropylethylamine; DMF = N,N-dimethylformamide; DMSO = dimethyl sulfoxide; DMA = N,N-dimethylacetamide; dppf = 1,1'-bis(diphenylphosphino)ferrocene); DTBPF = 1,1'-bis(di-tert-butylphosphino)ferrocene; EtOAc = ethyl acetate; equiv=equivalent; Ex = Example; h=time; KOAc = potassium acetate; LiHMDS = lithium bis(trimethylsilyl)amide [LiN(SiMe3)2]; mCPBA = metachloroperbenzoic acid; MeOH = methanol; NMP = N-methyl-2-pyrrolidone; min=minutes; Pd(dppf)Cl2 = [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II); RT=room temperature; satd.=saturated solution; TEA = triethylamine; TFA = trifluoroacetic acid; THF = tetrahydrofuran.

[0065] While the inventive concepts have been described in terms of exemplary principles and embodiments, those skilled in the art will recognize that changes may be made to, and equivalents substituted for, what has been described without departing from the scope and spirit of the present disclosure as defined by the following claims.

[0066] Example 1 2-((S)-1-Acryloyl-4-(7-(8-chloronaphthalen-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-1,5-naphthyridin-4-yl)piperazin-2-yl)acetonitrile (1b) [ka] Example 1(1b) was prepared as shown in Scheme 2 below. [ka]

[0067] 7-Bromo-4-chloro-1,5-naphthyridine-1-oxide (16). This compound was prepared as described on pages 64-65 of WO2020150114 by reacting mCPBA and 3-bromo-8-chloro-1,5-naphthyridine (15; CAS # 97267-61-3; 1.70 g, 7.02 mmol) in CHCl to give 1.50 g (83%) of 7-bromo-4-chloro-1,5-naphthyridine-1-oxide (16) as a pale yellow solid: HPLC-MS (ES) + ) m / z [M+H + ]=259, 261, 263; 1 H NMR (300 MHz, CDCl3) δ9.24 (d, J=2.2 Hz, 1H), 9.12 (d, J=2.2 Hz, 1H), 8.44 (d, J=6.7 Hz, 1H), 7.63 (d, J=6.7 Hz, 1H).

[0068] 7-Bromo-2,4-dichloro-1,5-naphthyridine (17). This compound was prepared as described on page 65 of WO2020150114. 7-Bromo-4-chloro-1,5-naphthyridine-1-oxide (16; 775 mg, 3.00 mmol) gave 750 mg (90%) of 7-bromo-2,4-dichloro-1,5-naphthyridine (17) as a pale pink solid: HPLC-MS (ES) + ) m / z [M+H + ]=277, 279, 281, 283; 1 H NMR (300 MHz, CDCl3) δ9.05 (d, J=2.1 Hz, 1H), 8.51 (d, J=2.1 Hz, 1H), 7.78 (s, 1H).

[0069] tert-Butyl (S)-4-(7-bromo-2-chloro-1,5-naphthyridin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (19). Triethylamine (1.90 mL, 13.6 mmol) was added to a suspension of (S)-2-(piperazin-2-yl)acetonitrile dihydrochloride (18; CAS # 1589082-26-7; 538 mg, 2.72 mmol) in anhydrous 1,4-dioxane (10 mL) and stirred at RT. After 5 h, the mixture was cooled to 0 °C, and anhydrous 1,4-dioxane (20 mL) was added, followed by the addition of 7-bromo-2,4-dichloro-1,5-naphthyridine (17; 750 mg, 2.72 mmol) in portions. After the addition was complete, the ice bath was removed and the mixture was allowed to warm to RT. After 5 min, the reaction mixture was heated to reflux for 23 h. The mixture was cooled to RT, and di-tert-butyl dicarbonate (1.87 mL, 8.16 mmol) was added. After 16 h, the mixture was diluted with EtOAc, washed with saturated aqueous NaCl (3X), dried (MgSO), filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with a gradient of 10% to 40% EtOAc in hexanes to afford 610 mg (48%) of tert-butyl (S)-4-(7-bromo-2-chloro-1,5-naphthyridin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (19) as a white solid: HPLC-MS (ES) + ) m / z [M+H + ]=466, 468, 470; 1 H NMR (300 MHz, CDCl3) δ8.89 (d, J=2.2 Hz, 1H), 8.37 (d, J=2.2 Hz, 1H), 6.83 (s, 1H), 4.94 (d, J=12.6 Hz, 1H), 4.63 (br s, 1H), 4.18 (br s, 1H), 3.76-3.68 (m, 1H), 3.33-3.06 (m, 4H), 2.79 (dd, J=5.3, 10.9 Hz, 1H), 1.53 (s, 9H).

[0070] tert-Butyl (S)-4-(7-bromo-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-1,5-naphthyridin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (21). A mixture of tert-butyl (S)-4-(7-bromo-2-chloro-1,5-naphthyridin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (19; 390 mg, 0.839 mmol), (2S)-1-methyl-2-pyrrolidinemethanol (20; CAS # 34381-71-0; 1.0 mL, 8.0 mmol), and CsCO (545 mg, 1.68 mmol) in anhydrous CHCN (8 mL) was heated to reflux. After 72 h, the mixture was cooled to RT, diluted with EtOAc, washed with saturated aqueous NaCl (3X), dried (MgSO), filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with a gradient of 0% to 15% MeOH in DCM to afford 80 mg (17%) of tert-butyl (S)-4-(7-bromo-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-1,5-naphthyridin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate as a light brown solid: HPLC-MS (ES) + ) m / z [M+H + ]=545, 547; 1H NMR (300 MHz、CDCl3) δ8.72 (d、J=2.2 Hz、1H)、8.22 (d、J=2.2 Hz、1H)、6.41 (s、1H)、4.74 (d、J=12.5 Hz、1H)、4.62 (br s、1H)、4.48 (dd、J=4.8、6.4 Hz、1H)、4.36 (dd、J=4.8、6.4 Hz、1H)、4.15 (br s、1H)、3.60 (d、J=5.8 Hz、1H)、3.22 (br s、1H)、3.32 (dd、J=7.7、8.8 Hz、1H)、3.14 (t、J=7.4 Hz、1H)、2.98 (td、J=3.3、9.1 Hz、2H)、2.83 (dd、J=5.6、10.7 Hz、1H)、2.67-2.57 (m、1H)、2.48 (s、3H)、2.34-2.22 (m、1H)、2.06-1.94 (m、1H)、1.93-1.71 (m、3H)、1.52 (s、9H)。

[0071] tert-Butyl (S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-1,5-naphthyridin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (23). A mixture of tert-butyl (S)-4-(7-bromo-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-1,5-naphthyridin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (21; 106 mg, 0.195 mmol), 2-(8-chloronaphthalen-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (22; 225 mg, 0.780 mmol), and KCO (487 mg, 3.53 mmol) in 1,4-dioxane (3 mL) and water (1.4 mL) was degassed by sparging with N with stirring for 30 min. Tetrakis(triphenylphosphine)palladium(0) (33 mg, 0.029 mmol) was added, and the reaction mixture was degassed by sparging with N with stirring for an additional 20 min. The reaction mixture was heated at 80 °C with stirring under a N atmosphere for 16 h. The reaction mixture was cooled to RT, diluted with EtOAc, and filtered through Celite, after which the filtrate was washed with saturated aqueous NaCl (3X), dried (MgSO), filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with a gradient of 0% to 10% MeOH in DCM to afford 48 mg (39%) of tert-butyl (S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-1,5-naphthyridin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (23) as an off-white solid: HPLC-MS (ES) + ) m / z [M+H + ]=627, 629; 1H NMR (300 MHz, CDCl3) δ8.67 (d, J=2.1 Hz, 1H), 8.02 (t, J=2.0 Hz, 1H), 7.94 (d, J=7.8 Hz, 1H), 7.87 (d, J=7.5 Hz, 1H), 7.55 (t, J=7.8 Hz, 2H), 7.42 (t, J=7.4 Hz, 2H), 6.45 (d, J=2.5 Hz, 1H), 4.92 (d, J=11.6 Hz, 1H), 4.79 (d, J=13.8 Hz, 1H), 4.67 (br s, 1H), 4.58-4.48 (m, 1H), 4.44-4.35 (m, 1H), 4.20 (br s, 1H), 3.70 (br t, J=10.3 Hz, 1H), 3.46-3.21 (m, 2H), 3.19-2.88 (m, 4H), 2.63 (br s, 1H), 2.49 (s, 3H), 2.34-2.22 (m, 1H), 2.08-1.69 (m, 3H), 1.53 (s, 9H).

[0072] 2-((S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-1,5-naphthyridin-4-yl)piperazin-2-yl)acetonitrile (24). A solution of tert-butyl (S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-1,5-naphthyridin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (23; 44 mg, 0.070 mmol) in CHCl (2 mL) was treated with a solution of 4 M HCl / 1,4-dioxane (0.2 mL) and stirred at RT. After 16 h, the mixture was treated with 0.1 M NaOH solution until the pH was basic. The layers were separated and the aqueous layer was extracted with CH2Cl2 (3X), dried (MgSO4), filtered, and concentrated under reduced pressure to give 31 mg (84%) of 2-((S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-1,5-naphthyridin-4-yl)piperazin-2-yl)acetonitrile (24) as an off-white solid: HPLC-MS (ES) + ) m / z [M+H +]=527、529; 1 H NMR (300 MHz、CDCl3) δ8.64(ddd、J=1.4、2.2、2.8 Hz、1H)、8.04-7.99 (m、1H)、7.95 (dd、J=1.2、7.0 Hz、1H)、7.88 (dd、J=1.2、7.0 Hz、1H)、7.62-7.49 (m、2H)、7.47-7.37 (m、2H)、6.46 (s、1H)、4.59-4.47 (m、1H)、4.44-4.32 (m、1H)、4.29-3.84 (m、2H)、3.80-3.73 (m、1H)、3.68-3.60 (m、1H)、3.57-3.44 (m、1H)、3.32-2.84 (m、4H)、2.74-2.56 (m、2H)、2.49 (s、3H)、2.43-2.20 (m、2H)、2.10-1.47 (m、4H)。

[0073] 2-((S)-1-Acryloyl-4-(7-(8-chloronaphthalen-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-1,5-naphthyridin-4-yl)piperazin-2-yl)acetonitrile (1b). A solution of 2-((S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-1,5-naphthyridin-4-yl)piperazin-2-yl)acetonitrile (24; 50 mg, 0.095 mmol) in CHCl (8 mL) was treated with EtN (16 μL, 0.114 mmol) and stirred at RT. The mixture was cooled to 0 °C and treated with acryloyl chloride (25; CAS # 814-68-6; 10 μL, 0.114 mmol). After 1 h at 0 °C, the mixture was diluted with CHCl and washed with HO (2X). The layers were separated, and the CHCl layer was dried (MgSO), filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with a gradient of 0% to 10% MeOH in DCM containing 5% NHOH (v / v) to afford 20 mg (36%) of 2-((S)-1-acryloyl-4-(7-(8-chloronaphthalen-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-1,5-naphthyridin-4-yl)piperazin-2-yl)acetonitrile (1b) as an off-white solid: HPLC-MS (ES) + ) m / z [M+H + ]=581, 583; 1H NMR (300 MHz, CDCl3) δ8.69 (bs, 1H), 8.04 (t, J=2.1 Hz, 1H), 7.96 (d, J=8.3 Hz, 1H), 7.89 (d, J=8.0 Hz, 1H), 7.62-7.05 (m, 2H), 7.47-7.38 (m, 2H), 6.65 (bs, 1H), 6.46 (d, J=2.8 Hz, 1H), 6.40 (dd, J=1.7, 18.2 Hz, 1H), 5.82 (d, J=10.5 Hz, 1H), 4.61-4.48 (m, 1H), 4.46-4.34 (m, 1H), 4.16-3.29 (m, 4H), 3.25-2.88 (m, 5H), 2.63 (bs, 1H), 2.49 (s, 3H), 2.39-2.18 (m, 1H), 2.10-1.68 (m, 5H).

[0074] Example 2 2-((S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-1,5-naphthyridin-4-yl)-1-(2-fluoroacryloyl)piperazin-2-yl)acetonitrile (1q) [ka] Example 2 (1q) was prepared as shown in Scheme 3 below. [ka] 2-((S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-1,5-naphthyridin-4-yl)-1-(2-fluoroacryloyl)piperazin-2-yl)acetonitrile (1q). A solution of 1-propanephosphonic anhydride (T3P, 0.2 mL, 0.275 mmol, 50% in EtOAc) was added to a solution of 2-((S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-1,5-naphthyridin-4-yl)piperazin-2-yl)acetonitrile (24; 29 mg, 0.055 mmol), 2-fluoroprop-2-enoic acid (26; CAS # 430-99-9; 14 mg, 0.154 mmol), and diisopropylethylamine (0.1 mL, 0.55 mmol) in 4 mL of EtOAc under a N2 atmosphere, and the mixture was stirred at RT. After 40 min, the reaction mixture was diluted with EtOAc and washed with saturated aqueous NaHCO3 (3X) and saturated aqueous NaCl (2X). The organic layer was then dried (MgSO), filtered, and concentrated under reduced pressure. The resulting crude product was purified by silica gel column chromatography eluting with a gradient of 10% to 100% EtOAc containing 1% EtN (v / v) in DCM to afford 17 mg (51%) of 2-((S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-1,5-naphthyridin-4-yl)-1-(2-fluoroacryloyl)piperazin-2-yl)acetonitrile (1q) as an off-white solid: HPLC-MS (ES) + ) m / z [M+H + ]=599,601; 1H NMR (300 MHz, CDCl3) δ8.69 (t, J=2.2 Hz, 1H), 8.04 (t, J=2.1 Hz, 1H), 7.96 (d, J=8.5 Hz, 1H), 7.89 (d, J=8.0 Hz, 1H), 7.59-7.50 (m, 2H), 7.47-7.37 (m, 2H), 6.46 (d, J=2.7 Hz, 1H), 5.41 (d, J=47.8 Hz, 1H), 5.24 (dd, J=3.9, 13.2 Hz, 1H), 4.60-4.47 (m, 1H), 4.45-4.33 (m, 1H), 3.87-3.72 (m, 1H), 3.50 (br s, 2H), 3.23-2.92 (m, 5H), 2.70-2.56 (m, 2H), 2.49 (s, 3H), 2.37-2.18 (m, 1H), 2.11-1.68 (m, 5H).

[0075] Example 3 1-(8-(8-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-6-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-3-yl)-2-fluoro-6-hydroxynaphthalen-1-yl)ethan-1-one (2aw) [ka] Example 3 (2aw) was prepared as shown in Scheme 4 below. [ka] tert-Butyl (1R,5S)-3-(7-bromo-2-chloro-1,5-naphthyridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (28). Triethylamine (1.80 mL, 12.5 mmol) was added to a suspension of 7-bromo-2,4-dichloro-1,5-naphthyridine (17; 1.15 g, 4.17 mmol) and tert-butyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylate (27; CAS # 149771-44-8; 0.97 g, 4.59 mmol) in anhydrous 1,4-dioxane (14 mL), and the mixture was heated at 90 °C under a N atmosphere. After 16 h, the mixture was cooled to RT, diluted with EtOAc, washed with saturated aqueous NaCl (3X), dried (MgSO), filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with a gradient of 1% to 25% EtOAc in hexanes to afford 520 mg (28%) of tert-butyl (1R,5S)-3-(7-bromo-2-chloro-1,5-naphthyridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (28) as a white solid: HPLC-MS (ES) + ) m / z [M+H + ]=453, 455, 457; 1 H NMR (300 MHz, CDCl3) δ8.75 (dd, J=1.1, 2.2 Hz, 1H), 8.18 (dd, J=1.1, 2.2 Hz, 1H), 6.71 (s, 1H), 4.39 (br s, 4H), 3.23 (br d, J=9.1 Hz, 1H), 2.14-1.94 (m, 5H), 1.50 (s, 9H).

[0076] tert-Butyl (1R,5S)-3-(7-bromo-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (30). To a solution of (2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methanol (29; CAS # 2097518-76-6; 151 mg, 0.953 mmol) in anhydrous DMF (6 mL) was added sodium hydride (38 mg, 0.953 mmol, 60% dispersion in mineral oil) under N2 atmosphere and cooled at 0 °C. After 35 min, tert-butyl (1R,5S)-3-(7-bromo-2-chloro-1,5-naphthyridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (28; 287 mg, 0.635 mmol) was added in one portion, and the mixture was allowed to warm to RT. After 16 h, the mixture was diluted with EtOAc, washed with saturated aqueous NaCl (4×), dried (MgSO), filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with a gradient of 0% to 10% MeOH in DCM to give 170 mg (46%) of tert-butyl (1R,5S)-3-(7-bromo-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (30) as a white foamy solid: HPLC-MS (ES) + ) m / z [M+H + ]=576, 578; 1H NMR (300 MHz、CDCl3) δ8.60 (d、J=2.2 Hz、1H)、8.16 (d、J=2.2 Hz、1H)、6.25 (s、1H)、5.27 (d、J=54.1 Hz、1H)、4.35 (br s、2H)、4.25 (d、J=10.5 Hz、1H)、4.13 (d、J=10.5 Hz、1H)、3.34-3.22 (m、2H)、3.19-2.93 (m、4H)、2.24-1.80 (m、10H)、1.68 (br s、2H)、1.49 (s、9H)。

[0077] tert-Butyl (1R,5S)-3-(7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)-naphthalen-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (32). tert-Butyl (1R,5S)-3-(7-bromo-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (30; 160 mg, 0.278 mmol), ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-1-yl)ethynyl)triisopropylsilane (31; CAS # 2621932-37-2; 168 mg, 0.328 mmol), and K2CO3 (156 mg, 1.14 mmol) were dissolved in 1,4-dioxane (3 mL) and water (0.5 mL). mL) and the mixture was degassed by sparging with N2 with stirring for 30 minutes. Tetrakis(triphenylphosphine)-palladium(0) (33.3 mg, 0.028 mmol) was added and the reaction mixture was degassed by sparging with N2 with stirring for an additional 20 minutes. The reaction mixture was heated to 80°C with stirring under an atmosphere of N2 for 16 hours, cooled to RT, diluted with EtOAc, and filtered through Celite. The filtrate was washed with saturated aqueous NaCl (2X), dried (MgSO4), filtered, and concentrated under reduced pressure.The crude product, a mixture of 32 and 33, was purified by silica gel column chromatography eluting with a gradient of 30% to 100% EtOAc in hexane to give 83 mg (34%) of tert-butyl (1R,5S)-3-(7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)-naphthalen-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (32) as an off-white solid (HPLC-MS (ES)) as a mixture of atropisomers. + ) m / z [M+H + ]=882) and 67 mg (33%) of 33 was obtained as a yellow-orange foamy solid (HPLC-MS (ES + ) m / z [M+H + ]=726).

[0078] tert-Butyl (1R,5S)-3-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (33). Tetrabutylammonium fluoride (0.1 mL, 0.10 mmol, 1 M in THF) was added to a solution of tert-butyl (1R,5S)-3-(7-(7-fluoro-3-(methoxy-methoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (32; 65 mg, 0.074 mmol) in anhydrous THF (1 mL), and the mixture was stirred at RT. After 1 h, the mixture was diluted with EtOAc, washed with saturated aqueous NaCl (3X), dried (MgSO4), filtered, and concentrated under reduced pressure to give 53 mg of tert-butyl (1R,5S)-3-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (33) as a yellow-orange foamy solid: HPLC-MS (ES) + ) m / z [M+H + ]=726; 1H NMR (300 MHz、CDCl3) δ9.04 (d、J=2.2 Hz、1H)、8.35 (d、J=2.2 Hz、1H)、7.88 (br d、J=1.0 Hz、1H)、7.78 (dd、J=4.1、4.7 Hz、1H)、7.29 (d、 J=9.1 Hz、1H)、6.29 (s、1H)、5.35 (s、2H)、5.30 (d、J=53.3 Hz、1H)、4.41 (br s、2H)、4.34 (d、J=10.5 Hz、1H)、4.20 (d、J=10.5 Hz、1H)、3.56 (s、3H)、3.37-2.94 (m、8H)、2.29-1.86 (m、10H)、1.63 (br s、2H)、1.50 (s、9H)。

[0079] 1-(8-(8-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-6-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-3-yl)-2-fluoro-6-hydroxynaphthalen-1-yl)ethan-1-one (2aw). Trifluoroacetic acid (1 mL) was added dropwise to a solution of tert-butyl (1R,5S)-3-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (33; 129 mg, 0.178 mmol) in dichloromethane (3 mL), and the mixture was stirred at RT under a N atmosphere. After 1 h, the mixture was diluted with DCM and slowly transferred via pipette to aqueous NHOH (25 mL) and stirred at RT. The layers were separated, the aqueous layer extracted with EtOAc (2X), and the combined organic layers were dried (MgSO), filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with a gradient of 0% to 20% MeOH in DCM containing 5% NH4OH (v / v) to give 16 mg (15%) of 1-(8-(8-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-6-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-3-yl)-2-fluoro-6-hydroxynaphthalen-1-yl)ethan-1-one (2aw) as a tan solid: HPLC-MS (ES) + ) m / z [M+H + ]=600; 1H NMR (300 MHz, DMSO-d6) δ10.92 (br s, 1H), 10.26 (s, 1H), 9.29 (br s, 1H), 9.11 (br s, 1H), 8.62 (d, J=2.0 Hz, 1H), 8.02 (dd, J=4.6, 6.0 Hz, 1H), 7.91 (d, J=2.0 Hz, 1H), 7.46 (t, J=9.3 Hz, 1H), 7.40 (d, J=2.3 Hz, 1H), 7.13 (d, J=2.0 Hz, 1H), 6.62 (s, 1H), 5.59 (d, J=52.4 Hz, 1H), 4.71-4.45 (m, 3H), 4.36 (d, J=12.3 Hz, 1H), 4.25 (br s, 2H), 4.07-3.65 (m, 4H), 2.37-2.01 (m, 10H), 1.99 (s, 3H).

[0080] Example 4 4-(8-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-6-((tetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-3-yl)naphthalen-2-ol (2a). [ka] Example 4(2a) was prepared as shown in Scheme 5 below.

[0081] [ka] tert-Butyl (1R,5S)-3-(7-bromo-2-((tetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (35). Sodium hydride (70 mg, 1.72 mmol, 60% w / w dispersion in mineral oil) was added to a solution of hexahydro-1H-pyrrolidin-7a-ylmethanol (34; CAS # 78449-72-6; 242 mg, 1.72 mmol) in anhydrous THF (8 mL) at 0 °C under a N atmosphere. After 30 min, tert-butyl (1R,5S)-3-(7-bromo-2-chloro-1,5-naphthyridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (28; 520 mg, 1.15 mmol) was added in one portion, and the mixture was heated to reflux. After 16 h, the mixture was cooled to RT, diluted with EtOAc, washed with saturated aqueous NaCl (3×), dried (MgSO), filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with a gradient of 0% to 10% MeOH in DCM containing 10% NH4OH (v / v) to give 300 mg (47%) of tert-butyl (1R,5S)-3-(7-bromo-2-((tetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (35) as a white solid: HPLC-MS (ES) + ) m / z [M+H + ]=558, 560; 1 H NMR (300 MHz, CDCl3) δ8.59 (d, J=2.2 Hz, 1H), 8.17 (d, J=2.2 Hz, 1H), 6.28 (s, 1H), 4.35 (br s, 2H), 4.22 (br s, 2H), 4.19 (s, 2H), 3.18-2.97 (m, 4H), 2.73-2.59 (m, 2H), 2.20-2.08 (m, 2H), 2.03-1.76 (m, 8H), 1.68-1.56 (m, 2H), 1.48 (s, 9H).

[0082] tert-Butyl (1R,5S)-3-(7-(3-hydroxynaphthalen-1-yl)-2-((tetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (37). A mixture of tert-butyl (1R,5S)-3-(7-bromo-2-((tetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (35; 290 mg, 0.520 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-2-ol (36; CAS # 2043962-01-0; 281 mg, 1.04 mmol), KCO (293 mg, 2.13 mmol) in 1,4-dioxane (5 mL) and water (1 mL) was degassed by sparging with N for 30 minutes. Tetrakis(triphenylphosphine)-palladium(0) (60 mg, 0.052 mmol) was added, and the reaction mixture was degassed by sparging with N for an additional 20 minutes. After sparging was complete, the reaction mixture was heated at 85° C. with stirring under an atmosphere of N for 16 hours. The reaction mixture was cooled to RT, diluted with EtOAc, and filtered through Celite. The organic layer was washed with saturated aqueous NaCl (3×), dried (MgSO), filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with a gradient of 0% to 10% MeOH in DCM containing 10% NH4OH (v / v) to afford 184 mg (57%) of tert-butyl (1R,5S)-3-(7-(3-hydroxynaphthalen-1-yl)-2-((tetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (37) as a tan solid: HPLC-MS (ES) + ) m / z [M+H + ]=622; 1H NMR (300 MHz、DMSO-d6) δ9.98 (br s、1H)、8.72 (d、J=2.2 Hz、1H)、8.02 (d、J=2.2 Hz、1H)、7.81 (br d、J=8.1 Hz、1H)、7.64 (br d、J=8.4 Hz、1H)、7.45 (br t、J=5.0 Hz、1H)、7.31-7.22 (m、2H)、7.14 (d、J=2.4 Hz、1H)、6.38 (s、1H)、4.35 (br d、J=11.1 Hz、2H)、4.27 (br s、2H)、4.06 (s、2H)、3.07 (br d、J=11.0 Hz、2H)、3.00-2.86 (m、2H)、2.60-2.50 (m、1H)、2.08 (br d、J=7.3 Hz、2H)、1.97-1.68 (m、9H)、1.63-1.49 (m、2H)、1.44 (s、9H)。

[0083] 4-(8-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-6-((tetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-3-yl)naphthalen-2-ol (2a). A solution of tert-butyl (1R,5S)-3-(7-(3-hydroxynaphthalen-1-yl)-2-((tetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (37; 172 mg, 0.277 mmol) in DCM (3 mL) was slowly treated dropwise with 4 M HCl in 1,4-dioxane (3 mL), and the resulting orange suspension was stirred at RT under a N atmosphere. After 2 h, the reaction was allowed to stand at RT. After 72 h, MeOH was added to the mixture, diluted with DCM, and the resulting solution was slowly transferred via pipette to NHOH(aq) and stirred at RT. The layers were separated, and the aqueous layer was extracted once with EtOAc and once with DCM. The combined organic layers were dried (MgSO), filtered, and concentrated under reduced pressure. The resulting crude product was purified by silica gel column chromatography eluting with a gradient of 0% to 15% MeOH in DCM containing 10% NHOH (v / v) to afford 102 mg (71%) of 4-(8-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-6-((tetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-3-yl)naphthalen-2-ol (2a) as a tan solid: HPLC-MS (ES) + ) m / z [M+H + ]=522; 1H NMR (300 MHz, DMSO-d6) δ10.0 (br s, 1H), 8.70 (d, J=2.2 Hz, 1H), 8.00 (d, J=2.2 Hz, 1H), 7.81 (br d, J=8.1 Hz, 1H), 7.64 (br d, J=8.4 Hz, 1H), 7.45 (br t, J=5.0 Hz, 1H), 7.32-7.21 (m, 2H), 7.14 (d, J=2.4 Hz, 1H), 6.26 (s, 1H), 4.29 (br d, J=9.5 Hz, 2H), 4.05 (s, 2H), 3.51 (br s, 2H), 3.32 (br s, 2H), 3.01 (br d, J=10.7 Hz, 2H), 2.97-2.87 (m, 2H), 2.60-2.50 (m, 1H), 2.04-1.64 (m, 10H), 1.62-1.48 (m, 2H).

[0084] Example 5 (S)-2-(1-Acryloyl-4-(7-(8-chloronaphthalen-1-yl)-2-((tetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)piperazin-2-yl)acetonitrile (1ae) [ka] Example 5 (1ae) was prepared as shown in Scheme 6 below. [ka] tert-Butyl (S)-4-(7-bromo-2-((tetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (38). Sodium hydride (142 mg, 3.54 mmol, 60% w / w dispersion in mineral oil) was added to a solution of hexahydro-1H-pyrrolidin-7a-ylmethanol (34; CAS # 78449-72-6; 500 mg, 3.54 mmol) in anhydrous THF (20 mL) at 0 °C under a N atmosphere. After 30 min, tert-butyl (S)-4-(7-bromo-2-chloro-1,5-naphthyridin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (19; 1.10 g, 2.36 mmol) was added in one portion and the mixture was heated to reflux. After 16 h, the mixture was cooled to RT, diluted with EtOAc, washed with saturated aqueous NaCl (3X), dried (MgSO), filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with a gradient of 0% to 6% MeOH in DCM containing 10% NH4OH (v / v) to afford 734 mg (54%) of tert-butyl (S)-4-(7-bromo-2-((tetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (38) as a white foam: HPLC-MS (ES) + ) m / z [M+H + ]=571, 573; 1H NMR (300 MHz、CDCl3) δ8.71(d、J=2.0 Hz、1H)、8.20 (d、J=2.2 Hz、1H)、6.40 (s、1H)、4.72 (d、J=12.5 Hz、1H)、4.61 (br s、1H)、4.21 (d、J=3.6 Hz、2H)、4.13 (br s、1H)、3.62 (d、J=4.3 Hz、1H)、3.43 - 3.18 (m、2H)、3.17 - 3.05 (m、2H)、3.04 - 2.91 (m、2H)、2.86 (dd、J=5.7、16.4 Hz、1H)、2.74 - 2.60 (m、2H)、2.08 - 1.77 (m、6H)、1.71 - 1.57 (m、2H)、1.53 (s、9H)。

[0085] tert-Butyl (S)-4-(7-(8-chloronaphthalen-1-yl)-2-((tetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (39). A mixture of tert-butyl (S)-4-(7-bromo-2-((tetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (38; 722 mg, 1.27 mmol), 2-(8-chloronaphthalen-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (22; CAS#: 2454397-84-1; 731 mg, 2.54 mmol), and KCO (718 mg, 5.21 mmol) in dioxane (12 mL) and water (2.4 mL) was degassed by sparging with N with stirring for 30 min. Tetrakis(triphenylphosphine)palladium(0) (147 mg, 0.127 mmol) was added and the reaction mixture was degassed by sparging with N2 with stirring for an additional 20 minutes. The reaction mixture was heated at 80°C with stirring under an atmosphere of N2 for 16 hours. The reaction mixture was cooled to RT, diluted with EtOAc, and filtered through Celite. The filtrate was washed with saturated aqueous NaCl (3X), dried (MgSO4), filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with a gradient of 0% to 6% MeOH in DCM containing 10% NH4OH (v / v) to give 362 mg (44%) of tert-butyl (S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-1,5-naphthyridin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (39) as a white foam: HPLC-MS (ES) + ) m / z [M+H + ]=653, 655; 1H NMR (300 MHz、CDCl3) δ8.66(t、J=2.2 Hz、1H)、8.01 (t、J=2.0 Hz、1H)、7.95 (d、J=8.1 Hz、1H)、7.88 (d、J=8.1 Hz、1H)、7.61 - 7.49 (m、2H)、7.47 - 7.37 (m、2H)、6.44 (d、J=3.2 Hz、1H)、4.86 (dd、J=12.5、40.0 Hz、1H)、4.68 (br s、1H)、4.36 - 3.96 (m、2H)、3.87 - 3.56 (m、1H)、3.49 - 3.20 (m、2H)、3.18 - 2.83 (m、6H)、2.77 - 2.54 (m、2H)、2.10 - 1.77 (m、6H)、1.71 - 1.57 (m、2H)、1.53 (s、9H)。

[0086] (S)-2-(4-(7-(8-chloronaphthalen-1-yl)-2-((tetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)piperazin-2-yl)acetonitrile (40). A solution of tert-butyl (S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((S)-1-methylpyrrolidin-2-yl)methoxy)-1,5-naphthyridin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (39; 349 mg, 0.534 mmol) in CHCl (10 mL) was slowly treated dropwise with 4 M HCl / dioxane (3.3 mL) resulting in the formation of an orange sticky solid, and the mixture was stirred at RT. After 3 h, the solid was sampled and determined to be the product by LC / MS. The DCM layer was decanted off, and the sticky solid was dissolved in MeOH. The solution was diluted with saturated aqueous NaHCO3, extracted with DCM (5X), dried (MgSO4), filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with a gradient of 0% to 10% MeOH in DCM containing 10% NH4OH (v / v) to afford 191 mg (65%) of (S)-2-(4-(7-(8-chloronaphthalen-1-yl)-2-((tetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)piperazin-2-yl)acetonitrile (40) as a white foam: HPLC-MS (ES) + ) m / z [M+H + ]=554, 556; 1H NMR (300 MHz, CDCl3) δ8.63 (d, J=2.1 Hz, 1H), 8.00 (d, J=2.0 Hz, 1H), 7.96 (d, J=8.1 Hz, 1H), 7.88 (d, J=8.1 Hz, 1H), 7.60 - 7.49 (m, 2H), 7.47 - 7.36 (m, 2H), 6.45 (s, 1H), 4.23 (d, J=4.8 Hz, 2H), 3.95 (dd, J=11.4, 33.2 Hz, 1H), 3.60 - 3.39 (m, 1H), 3.34 - 2.96 (m, 6H), 2.76 - 2.50 (m, 4H), 2.12 - 1.73 (m, 8H), 1.71 - 1.50 (m, 2H).

[0087] (S)-2-(1-Acryloyl-4-(7-(8-chloronaphthalen-1-yl)-2-((tetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)piperazin-2-yl)acetonitrile (1ae). A solution of (S)-2-(4-(7-(8-chloronaphthalen-1-yl)-2-((tetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)piperazin-2-yl)acetonitrile (40; 83 mg, 0.15 mmol) in CHCl (12 mL) was treated with EtN (25 μL, 0.17 mmol). The mixture was cooled to 0 °C, and acryloyl chloride (25; CAS # 814-68-6; 15 μL, 0.17 mmol) was added, and the mixture was stirred in an ice bath. After 1.5 h, the mixture was diluted with CHCl, washed with HO (2X), dried (MgSO), filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluting with a gradient of 0% to 5% MeOH in DCM containing 10% NHOH (v / v)) to afford 45 mg (49%) of (S)-2-(1-acryloyl-4-(7-(8-chloronaphthalen-1-yl)-2-((tetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)piperazin-2-yl)acetonitrile (1ae) as an off-white powder: HPLC-MS (ES) + ) m / z [M+H+ ]=607, 609; 1 H NMR (300 MHz, CDCl3) δ8.68 (br t, J=1.9 Hz, 1H), 8.02 (t, J=1.9 Hz, 1H), 7.96 (d, J=8.2 Hz, 1H), 7.88 (d, J=8.0 Hz, 1H), 7.62 - 7.49 (m, 2H), 7.47 - 7.35 (m, 2H), 6.64 (br s, 1H), 6.45 (d, J=2.9 Hz, 1H), 6.40 (dd, J=1.4, 16.8 Hz, 1H), 5.82 (d, J=10.3 Hz, 1H), 5.36 - 4.42 (m, 2H), 4.36 - 4.14 (m, 2H), 4.12 - 3.26 (m, 3H), 3.24 - 2.81 (m, 6H), 2.75 - 2.52 (m, 2H), 2.10 - 1.52 (m, 8H).

[0088] Example 6 (S)-2-(4-(7-(8-chloronaphthalen-1-yl)-2-((tetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-1-(2-fluoroacryloyl)piperazin-2-yl)acetonitrile (1af) [ka] Example 6 (1af) was prepared as shown in Scheme 7 below. [ka] (S)-2-(4-(7-(8-chloronaphthalen-1-yl)-2-((tetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-1-(2-fluoroacryloyl)piperazin-2-yl)acetonitrile (1af). 1-Propanephosphonic anhydride solution (T3P, 0.48 mL, 0.75 mmol, 50% in EtOAc) was added to a mixture of (S)-2-(4-(7-(8-chloronaphthalen-1-yl)-2-((tetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)piperazin-2-yl)acetonitrile (40; 83 mg, 0.15 mmol), 2-fluoroprop-2-enoic acid (26; CAS # 430-99-9; 41 mg, 0.45 mmol), and diisopropylethylamine (0.27 mL, 1.50 mmol) in 11 mL of EtOAc under a N2 atmosphere, and the mixture was stirred at RT. After 40 min, additional 1-propanephosphonic anhydride solution (T3P, 0.3 mL) was added, and the mixture was stirred at RT. After 1.5 h, the reaction mixture was diluted with EtOAc, washed with 5% aqueous NaHCO (3X), dried (MgSO), filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluting with a gradient of 10% to 100% EtOAc containing 5% EtN (v / v)) to afford 25 mg (27%) of (S)-2-(4-(7-(8-chloronaphthalen-1-yl)-2-((tetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-1-(2-fluoroacryloyl)piperazin-2-yl)acetonitrile (1af) as an off-white powder: HPLC-MS (ES) + ) m / z [M+H + ]=625, 627; 1H NMR (300 MHz, CDCl3) δ8.68 (t, J=2.2 Hz, 1H), 8.02 (t, J=2.1 Hz, 1H), 7.96 (d, J=8.2 Hz, 1H), 7.88 (d, J=8.0 Hz, 1H), 7.61 - 7.49 (m, 2H), 7.47 - 7.36 (m, 2H), 6.45 (d, J=3.2 Hz, 1H), 5.41 (br d, J=48.4 Hz, 1H), 5.25 (dd, J=3.6, 16.9 Hz, 1H), 5.15 - 4.41 (m, 2H), 4.35 - 4.14 (m, 2H), 3.81 (br d, J=9.4 Hz, 1H), 3.72 - 3.29 (m, 2H), 3.21 - 2.90 (m, 5H), 2.77 - 2.55 (m, 2H), 2.09 - 1.78 (m, 7H), 1.72 - 1.57 (m, 2H).

[0089] Example 7 2-((S)-1-Acryloyl-4-(7-(8-chloronaphthalen-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)piperazin-2-yl)acetonitrile (1ag) [ka] Example 7 (1ag) was prepared as shown in Scheme 8 below. [ka] tert-Butyl (S)-4-(7-bromo-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (41). Sodium hydride (142 mg, 3.54 mmol, 60% w / w dispersion in mineral oil) was added to a solution of (2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methanol (29; CAS # 2097518-76-6; 563 mg, 3.54 mmol) in anhydrous THF (20 mL) at 0 °C under a N atmosphere. After 30 min, tert-butyl (S)-4-(7-bromo-2-chloro-1,5-naphthyridin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (19; 1.10 g, 2.36 mmol) was added in one portion, the cooling bath was removed, and the mixture was stirred at RT. After 30 min, the mixture was heated to reflux. After 16 h, the mixture was cooled to RT, diluted with EtOAc, washed with saturated aqueous NaCl (3X), dried (MgSO), filtered, and concentrated under reduced pressure. The crude solid was suspended in EtO and stirred at RT. After 16 h, the solid was filtered, washed with EtO, and dried to give 900 mg (65%) of tert-butyl (S)-4-(7-bromo-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (41) as an off-white solid: HPLC-MS (ES + ) m / z [M+H + ]=589, 591; 1H NMR (300 MHz、CDCl3) δ8.72 (d、J=2.2 Hz、1H)、8.19 (d、J=2.2 Hz、1H)、6.38 (s、1H)、5.28 (br d、J=54.1 Hz、1H)、4.73 (d、J=12.7 Hz、1H)、4.62 (br s、1H)、4.29 (d、J=10.6 Hz、1H)、4.14 (d、J=10.6 Hz、2H)、3.62 (br d、J=11.3 Hz、1H)、3.39 - 3.10 (m、5H)、3.05 - 2.90 (m、3H)、2.84 (dd、J=5.6、16.4 Hz、1H)、2.21 (br d、J=2.9 Hz、1H)、2.14 - 1.79 (m、5H)、1.52 (s、9H)。

[0090] tert-Butyl (S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (42). A mixture of tert-butyl (S)-4-(7-bromo-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (41; 800 mg, 1.36 mmol), 2-(8-chloronaphthalen-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (22; CAS#: 2454397-84-1; 783 mg, 2.72 mmol), and KCO (563 mg, 4.08 mmol) in dioxane (7 mL) and water (4 mL) was degassed by sparging with N with stirring for 20 min. Tetrakis(triphenylphosphine)palladium(0) (78 mg, 0.068 mmol) was added, and the reaction mixture was degassed by sparging with N2 with stirring for an additional 20 minutes. The reaction mixture was heated at 80°C with stirring under an atmosphere of N2 for 16 hours. The reaction mixture was cooled to RT and diluted with EtOAc. The filtrate was washed with saturated aqueous NaCl (3X), dried (MgSO4), filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluting with a gradient of 0% to 5% MeOH in DCM) to give 400 mg (44%) of tert-butyl (S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (42) as a beige solid: HPLC-MS (ES) + ) m / z [M+H + ]=671, 673; 1H NMR (300 MHz、CDCl3) δ8.67 (t、J=2.4 Hz、1H)、8.01 (dd、J=1.3、2.0 Hz、1H)、7.95 (d、J=8.2 Hz、1H)、7.88 (dd、J=1.2、8.1 Hz、1H)、7.60 - 7.49 (m、2H)、7.47 - 7.36 (m、2H)、6.42 (d、J=3.7 Hz、1H)、5.29 (br d、J=53.7 Hz、1H)、4.86 (dd、J=12.3、40.2 Hz、1H)、4.68 (br s、1H)、4.33 (dd、J=7.8、10.6 Hz、1H)、4.17 (t、J=9.7 Hz、2H)、3.72 (br t、J=11.4 Hz、1H)、3.49 - 3.14 (m、5H)、3.10 - 2.83 (m、4H)、2.32 - 1.64 (m、6H)、1.52 (s、9H)。

[0091] 2-((S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)piperazin-2-yl)acetonitrile (43). A solution of tert-butyl (S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (42; 388 mg, 0.58 mmol) in CHCl (10 mL) was slowly treated dropwise with 4 M HCl / dioxane (3.6 mL) under a N atmosphere, resulting in the formation of a sticky solid, and the mixture was stirred at RT. After 2.5 h, the solid was sampled and determined to be the product by LC / MS. The DCM layer was decanted off, and the sticky solid was dissolved in MeOH. The solution was diluted with saturated aqueous NaHCO and extracted with DCM (3X), dried (MgSO), filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with a gradient of 0% to 10% MeOH in DCM containing 10% NH4OH (v / v) to give 198 mg (60%) of 2-((S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)piperazin-2-yl)acetonitrile (43) as an off-white solid: HPLC-MS (ES) + ) m / z [M+H + ]=571, 573; 1H NMR (300 MHz、DMSO-d6) δ8.58 (t、J=2.2 Hz、1H)、8.16 (dd、J=1.1、8.2 Hz、1H)、8.10 (dd、J=1.2、8.1 Hz、1H)、7.93 (t、J=1.9 Hz、1H)、7.73 - 7.62 (m、2H)、7.61 - 7.50 (m、2H)、6.42 (s、1H)、5.29 (br d、J=54.1 Hz、1H)、4.36 - 4.17 (m、1H)、4.08 (dd、J=10.4、26.5 Hz、3H)、3.23 - 2.60 (m、12H)、2.25 - 1.68 (m、6H)。

[0092] 2-((S)-1-Acryloyl-4-(7-(8-chloronaphthalen-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)piperazin-2-yl)acetonitrile (1ag). A solution of 2-((S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)piperazin-2-yl)acetonitrile (43; 90 mg, 0.16 mmol) in CHCl (12 mL) was treated with EtN (25 μL, 0.17 mmol). The mixture was cooled to 0 °C, acryloyl chloride (25; CAS # 814-68-6; 15 μL, 0.17 mmol) was added, and the mixture was stirred in an ice bath. After 1.5 h, the mixture was cooled to 0 °C again, and an additional 6.8 μL of EtN and 3.8 μL of acryloyl chloride were added and stirred in an ice bath. After 1 h, the mixture was diluted with CHCl, washed with HO (2X), dried (MgSO), filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluting with a gradient of 0% to 10% MeOH in DCM) to give 73 mg (74%) of 2-((S)-1-acryloyl-4-(7-(8-chloronaphthalen-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)piperazin-2-yl)acetonitrile (1ag) as an off-white solid: HPLC-MS (ES + ) m / z [M+H + ]=625, 627; 1H NMR (300 MHz, CDCl3) δ8.69 (br t, J=2.1 Hz, 1H), 8.02 (dd, J=1.0, 2.1 Hz, 1H), 7.96 (dd, J=1.1, 8.2 Hz, 1H), 7.88 (dd, J=1.2, 8.1 Hz, 1H), 7.59 - 7.50 (m, 2H), 7.47 - 7.38 (m, 2H), 6.63 (br s, 1H), 6.46 - 6.33 (m, 2H), 5.82 (d, J=10.6 Hz, 1H), 5.29 (br d, J=53.2 Hz, 1H), 5.06 - 4.45 (m, 2H), 4.35 (dd, J=7.1, 10.4 Hz, 1H), 4.19 (dd, J=9.2, 10.4 Hz, 1H), 4.12 - 3.52 (m, 2H), 3.42 - 2.80 (m, 8H), 2.46 - 1.77 (m, 7H).

[0093] Example 8 2-((S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-1-(2-fluoroacryloyl)piperazin-2-yl)acetonitrile (1ah) [ka] Example 8 (1ah) was prepared as shown in Scheme 9 below. [ka] 2-((S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-1-(2-fluoroacryloyl)piperazin-2-yl)acetonitrile (1ah). A solution of 1-propanephosphonic anhydride (T3P, 0.52 mL, 0.80 mmol, 50% in EtOAc) was added to a mixture of 2-((S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)piperazin-2-yl)acetonitrile (43; 83 mg, 0.15 mmol), 2-fluoroprop-2-enoic acid (26; CAS # 430-99-9; 41 mg, 0.45 mmol), and Et3N (0.12 mL, 0.80 mmol) in 10 mL of EtOAc under a N2 atmosphere. The mixture was stirred at RT, forming a light pink reaction mixture. After 1 h, the light pink color changed to gold. The mixture was stirred at RT, and after 3 h the reaction mixture was light brown. The mixture was diluted with EtOAc, washed with 5% aqueous NaHCO3 (3X), dried (MgSO4), filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluting with a gradient of 0% to 10% MeOH in DCM) to afford 36 mg (39%) of 2-((S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-1-(2-fluoroacryloyl)piperazin-2-yl)acetonitrile (1ah) as an off-white solid: HPLC-MS (ES) + ) m / z [M+H + ]=643, 645; 1H NMR (300 MHz, CDCl3) δ8.69 (t, J=2.2 Hz, 1H), 8.02 (dd, J=1.2, 1.9 Hz, 1H), 7.96 (dd, J=1.0, 8.1 Hz, 1H), 7.88 (dd, J=1.0, 8.1 Hz, 1H), 7.59 - 7.50 (m, 2H), 7.47 - 7.38 (m, 2H), 6.42 (d, J=3.7 Hz, 1H), 5.41 (br d, J=47.5 Hz, 1H), 5.29 (br d, J=53.2 Hz, 1H), 5.25 (dd, J=3.8, 17.0 Hz, 1H), 5.02 (br s, 1H), 4.87 (br s, 1H), 4.35 (dd, J=7.4, 10.6 Hz, 1H), 4.19 (dd, J=9.2, 10.3 Hz, 1H), 3.82 (br d, J=10.2 Hz, 1H), 3.54 (br s, 1H), 3.39 - 2.88 (m, 7H), 2.28 - 1.78 (m, 8H).

[0094] Nucleotide exchange assay The biological activity of the present examples was measured in a KRAS G12D / SOS1 nucleotide exchange assay performed by Reaction Biology Corporation (RBC), 1 Great Valley Parkway, Suite 2, Malvern, PA 19355, USA. This assay evaluates the SOS1-mediated Bodipy-GDP to GTP exchange observed with KRAS G12C and KRAS G12D.

[0095] Compounds were assayed at 10 concentrations of IC with 3-fold serial dilutions at a starting concentration of 5 μM for ARS1620 and 10 μM for Examples 1-4, MRTX849 and MRTX1133. 50 The compound pre-incubation time was 30 minutes at room temperature, and curve fitting was performed if the activity at the highest compound concentration was less than 65%. Reaction buffer: 40mM HEPES 7.4, 10mM MgCl2, 1mM DTT 0.002% Triton X100, 0.5% DMSO. Enzyme: SOS1 (RBC cat# MSC-11-502). Recombinant human SOS1 (Genbank accession# NM_033360.3; aa 564-1049, expressed in E. coli with C-terminal Strep II). KRAS G12C and KRAS G12D: Recombinant human KRAS G12C or KRAS G12D (aa 2-169, expressed in E. coli and with an N-terminal TEV-cleavable his-tag) was incubated with 5-fold excess Bodipy TM - Preloaded with GDP and excess Bodipy TM -GDP is removed from the loaded protein using a spin desalting column. Final concentration: KRAS-Bodipy TM -GDP is 0.125 μM; SOS1 is 750 nM; and GTP is 25 μM.

[0096] Reaction Method: 1. Inject 10 μL of freshly prepared 1.5x KRAS solution in reaction buffer into the reaction well. 2. Compounds in 100% DMSO are delivered into the buffer solution using acoustic technology (Echo550; nanoliter range). 3. Incubate the compound with KRAS for 30 minutes at room temperature. 4. Prepare a 3x (SOS1+GTP) solution in reaction buffer. 5. Inject 5 μL of SOS1+GTP solution into the reaction wells (as a control without SOS1, inject only GTP into column 1). 6. Monitor the progress of the reaction by the decrease in fluorescent signal using a PHERAstar (BMG Labtech plate reader (Ex / Em=485 / 520)) for 30 minutes at room temperature.

[0097] Data analysis: Fluorescence data were normalized using the following formula and fitted to a "one-phase exponential decay" equation using GraphPad Prism software. The plateau was fixed at zero (used for non-covalent inhibitors) and the ratio x 1000 was calculated as the IC 50was used to calculate the value.

number

[0098] Background-subtracted signals (wells without SOS1 protein were used as background) were converted to % activity relative to the DMSO control. Data were analyzed using GraphPad Prism 4 with a 4-parameter "sigmoidal dose-response (variable slope)" with a Hill slope. Constraints were a lower limit (constant equal to 0) and an upper limit (must be less than 120).

[0099] result: [Table 1] * I C 50 Values ​​were calculated using the dRFU analysis method for covalent inhibitors, using Bodipy-GDP / KRAS G12C as the substrate and adding 0.5% DMSO to the reaction mixture. ARS-1620 and MRTX-849 are standards for KRAS G12C. [Table 2] * I C 50 The values ​​were analyzed using Bodipy-GDP / KRAS G12D as the substrate and 0.5% DMSO in the reaction mixture using the reversible inhibitor reaction rate constant method (plateau = 0). MRTX1133 is a KRAS G12D standard sample.

[0100] KRAS G12C cell assay KRAS G12C cellular activity was measured using a target engagement cellular assay (NanoBRET) in transiently transfected HEK293 cells by Reaction Biology Corporation (RBC), 1 Great Valley Parkway, Suite 2, Malvern, PA 19355, USA. TM ) were measured. HEK293 cells were cultured to 70-80% confluence prior to the assay, then trypsinized and harvested. BI-2852 was used as a KRAS G12C standard compound. Prior to the assay, each test compound solution was transferred from the compound source plate to the wells of a 384-well white non-binding surface plate using an Echo 550.

[0101] A 10 μg / mL solution of DNA in serum-free Opti-MEM was prepared, consisting of 1 μg of LgBiT®-KRAS (G12C)-NanoLuc fusion vector, 1 μg of SmBiT®-KRAS (G12C)-NanoLuc fusion vector, and 8 μg of transfection carrier DNA. This mixture was then treated with 30 μL of FuGENE HD Transfection Reagent per milliliter of DNA mixture to form lipid:DNA complexes. The resulting mixture was mixed by gentle inversion and incubated at ambient temperature for 20 minutes to allow complex formation. A mixture of 1 part lipid:DNA complex and 20 parts suspended HEK293 cells was added to a sterile conical tube and mixed by gentle inversion. The cell+lipid:DNA complex mixture was then added to a sterile tissue culture dish and incubated for 24 hours. The medium was removed from the dish by aspiration, followed by trypsinization to release the cells from the tissue culture dish. The trypsin was then neutralized with serum-containing medium and the cells were pelleted in the conical tube by centrifugation at 200 × g for 5 min at a cell density of 2 × 10 in Opti-MEM without phenol red. 5 Complete 20X NanoBRET TMOne portion of the RAS Tracer Reagent was dispensed into 20 portions of cells in the conical tube and gently inverted to mix. The resulting cell suspension was dispensed into a white 384-well NBS plate containing test compounds (starting at 10 μM, with 10 three-fold dilutions) and incubated at 37°C, 5% CO2 for 2 hours. The final concentration of RAS tracer K2 was 1 μM. The NBS plate was removed from the incubator and allowed to equilibrate to room temperature for 15 minutes.

[0102] Freshly prepared substrate solution (3X) in assay medium was added to each well of a 384-well NBS plate and incubated at room temperature for 3 minutes. The donor emission wavelength (460 nm) and acceptor emission wavelength (600 nm) were measured using an Envision 2104 plate reader. For each sample, the raw BRET ratio was calculated by dividing the acceptor emission value (600 nm) by the donor emission value (460 nm). To correct for background, the BRET ratio in the absence of tracer (average value of the no-tracer control samples) was subtracted from the BRET ratio of each sample. The BRET ratio was calculated using the following formula: BRET ratio = [(acceptor sample ÷ donor sample) - (acceptor no-tracer control ÷ donor no-tracer control)]. The normalized BRET response (%) was calculated using the following formula: (BRET ratio of test compound / BRET ratio of DMSO control) * 100%. IC 50 Plot the curve and calculate the IC in GraphPad Prism 4 based on the sigmoidal dose-response equation. 50 The value was calculated.

[0103] result: [Table 3] * NanoBRET TM Target engagement cell assay (KRAS G12C). BI-2852 is the KRAS G12C standard.

[0104] CellTiter-Glo Viability Assay Protocol material: The standard compound staurosporine was purchased from Sigma-Aldrich (Saint Louis, MI). CellTiter-Glo® 2.0 Luminescent Cell Viability Assay Reagent was purchased from Promega (Madison, WI). The MIA PaCa-2 cell line was purchased from the American Type Culture Collection (Manassas, VA). MIA PaCa-2 cells were cultured in DMEM containing 10% FBS, 2.5% horse serum, 100 μg / ml penicillin, and 100 μg / ml streptomycin. Cultures were maintained at 37°C in a humidified atmosphere of 5% CO2 and 95% air.

[0105] procedure: 1. Test compounds and the standard compound staurosporine were diluted 10 times 3-fold in DMSO solution in the source plate, starting from 10 mM. 2.25 nL of test compound or 25 nL of staurosporine was transferred by the Echo 550 from the source plate to each well of a 384-well cell culture plate. 3. 25 μL of medium containing 2000 cells was added in duplicate to each well of the cell culture plate. 4. Cells were incubated with compounds for 72 hours at 37°C, 5% CO2. 5.25 μL of CellTiter-Glo 2.0 reagent was added to each well. 6. The contents were mixed on an orbital shaker for 2 minutes and incubated at room temperature for 15 minutes to allow the luminescent signal to stabilize. 7. Luminescence was recorded using an Envision 2104 Multilabel Reader (PerkinElmer, Santa Clara, CA). The number of viable cells in the culture was determined based on the quantification of ATP present in each culture well. 8. IC50 curves were plotted and IC50 values ​​were calculated using GraphPad Prism 4 based on the sigmoidal dose-response equation.

[0106] result: Table 4 Table 5

Claims

1. Formula I: 【Chemistry 1】 [During the ceremony, A is hydrogen, halogen, hydroxy, -C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 Alkynyl, -C 0-3 Alkyl (C 3-6 cycloalkyl), -C 1-6 Alkyl (halo), -C 1-6 Alkyl (OH), -O(C 1-4 alkyl), -C 1-3 Alkyl (C 1-4 Alkoxy), -CN, -CO 2 R 4 , -CO 2 N(R 4 ) 2 , -NO 2 , -N(R 4 ) 2 , -P(O)(R 5 ) 2 , -SR 4 , -S(O)R 4 , -SO 2 R 4 or aryl or heteroaryl optionally substituted with one or more 5- to 6-membered heterocyclic rings; Y and G may be the same or different and are hydrogen, halogen, C 1-4 Alkyl, C 1-4 Perdeuteroalkyl, -(C 0-2 alkyl)alkenyl, -(C 0-2 alkyl)alkynyl, -(C 0-2 alkyl)cycloalkyl, -C 1-4 Haloalkyl, -O(C 1-4 alkyl), -S(C 1-4 alkyl), -(C 0-2 alkyl) cyano, -O(C 1-4 haloalkyl) or -S(C 1-4 haloalkyl); L is a bond, O, S, or NR 4 and; m is 0 to 2; n is 0 to 2; Z is C(R 4 ) 2 , or C 3-7 a cyclic compound selected from cycloalkyl, a saturated or partially unsaturated 4- to 7-membered nitrogen-containing ring, and a saturated or partially unsaturated 7- to 10-membered nitrogen-containing bridged bicyclic ring; R 1 is hydrogen, hydroxy, halogen, -C 1-3 Alkyl, -C 1-3 Alkyl (OH), -C 1-3 Alkyl (halo), -C 1-3 Alkyl (C 1-3 alkoxy), -C 1-3 Alkyl (CN) or -C 1-3 Alkyl (P(O)R 5 2 ) are selected from; R 2 is selected from hydrogen, —C(O)CH═CH, —C(O)CF═CH or —C(O)CCl═CH, with the proviso that R 2 is hydrogen, m is 1 or 2; R 3 is hydrogen, halogen, hydroxy, -C 1-4 Alkyl, -C 2-4 Alkenyl, -C 2-4 Alkynyl, -C 0-3 Alkyl (C 3-6 cycloalkyl), -C 1-4 Alkyl (halo), -C 1-4 Alkyl (OH), -O(C 1-4 alkyl), -C 1-3 Alkyl (C 1-3 Alkoxy), -CN, -CO 2 R 4 , -CO 2 N(R 4 ) 2 , -NO 2 , -N(R 4 ) 2 , -PO(R 5 ) 2 , -SR 4 , -S(O)R 4 , -SO 2 R 4 or -(C 0-3 alkyl)R 6 Selected from; R 4 is hydrogen, C 1-4 selected from alkyl, aryl, or heteroaryl; R 5 is hydrogen, hydroxy, C 1-4 Alkyl, aryl, heteroaryl, C 1-4 selected from alkoxy, aryloxy, or heteroaryloxy; R 6 is N(R 4 ) 2 or a 4- to 7-membered saturated or unsaturated heterocyclic ring containing one or more heteroatoms selected from the group N, O and S. or a pharmaceutically acceptable salt, solvate or prodrug thereof.

2. The compound of claim 1, wherein the compound of formula I is selected from compounds 1a-1ah and 2a-2aw and pharmaceutically acceptable salts, solvates or prodrugs thereof: 【Chemistry 2】 【Transformation 3】 【Chemistry 4】 【Transformation 5】 【Transformation 6】 【Transformation 7】

3. 10. A pharmaceutical composition comprising a compound of claim 1 or 2, or a salt, solvate or prodrug thereof, together with a pharmaceutically acceptable carrier.

4. 1. A method of treating a disease, disorder, or medical condition in a patient, comprising: The method includes providing a therapeutic agent to a patient in need of treatment; The therapeutic agent is a compound according to any one of claims 1 to 3 or a salt, solvate or prodrug thereof. The method.

5. 5. The method of treating a disease, disorder, or medical condition of claim 4, wherein the disease comprises various cancers.

6. 6. The method of treating a disease, disorder or medical condition of claim 5, wherein the disease, disorder or medical condition is KRAS-mediated.

7. 7. The method of treating a disease, disorder or medical condition according to claim 6, wherein the disease, disorder or medical condition is mediated by KRAS mutant G12C or G12D.

8. 6. The method of treating a disease, disorder, or medical condition of claim 5, wherein the cancer is selected from glioma (glioblastoma), acute myelogenous leukemia, acute myeloid leukemia, myelodysplastic / myeloproliferative neoplasm, sarcoma, chronic myelomonocytic leukemia, non-Hodgkin's lymphoma, astrocytoma, melanoma, non-small cell lung cancer, small cell lung cancer, cholangiocarcinoma, chondrosarcoma, colon cancer, colorectal cancer, rectal cancer, or pancreatic cancer.

9. The method of any one of claims 4 to 8, further comprising administering at least one additional therapeutic agent to a patient in need of treatment.

10. 10. The method of claim 9, wherein the additional therapeutic agent is selected from doxorubicin, paclitaxel, docetaxel, cisplatin, camptothecin, temozolomide, Avastin, Herceptin, Erbitux, an EGFR inhibitor, osimertinib, rezivertinib, a CDK4 / 6 inhibitor, abemaciclib, palbociclib, ribociclib, a c-MET inhibitor, capmatinib, volitinib, an ALK inhibitor, crizotinib, alectinib, ceritinib, brigatinib, entrectinib, lorlatinib, a PD-1 antagonist, a PD-L1 antagonist, ipilimumab, embrolizumab, or nivolumab.

11. The compounds of formula I are compounds 1ae-1ah, 1b, 1d, 1i, 1q, 1s, 1x, 2z, 2al, 2an, 2ap and 2aq: 【Transformation 8】 or a salt, solvate or prodrug thereof.

12. 4. The pharmaceutical composition of claim 3, comprising any of compounds 1ae-1ah, 1b, 1q, or 2z, or a salt, solvate, or prodrug thereof, together with a pharmaceutically acceptable carrier.

13. 13. The method of claim 4, comprising providing the pharmaceutical composition of claim 12 to a patient in need of treatment.

14. 14. The method of claim 13, wherein the disease comprises various cancers.

15. 15. The method of claim 14, wherein the disease, disorder, or medical condition is KRAS-mediated.

16. 16. The method of claim 15, wherein the disease, disorder or medical condition is mediated by KRAS mutant G12C or G12D.

17. 15. The method of claim 14, wherein the cancer is selected from glioma (glioblastoma), acute myeloid leukemia, acute myelogenous leukemia, myelodysplastic / myeloproliferative neoplasm, sarcoma, chronic myelomonocytic leukemia, non-Hodgkin's lymphoma, astrocytoma, melanoma, non-small cell lung cancer, small cell lung cancer, cholangiocarcinoma, chondrosarcoma, colon cancer, colorectal cancer, rectal cancer, or pancreatic cancer.

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