1, 5-naphthyridine derivatives as KRAS oncoprotein inhibitors

Abstract

The present invention relates to inhibitors of Kirsteen rat sarcoma virus (KRAS) oncoprotein, and more particularly to certain 1, 5-naphthyridine derivatives of Formula I, as well as compositions comprising Formula I and methods of using compounds of Formula I to treat or prevent diseases, disorders or medical conditions mediated by KRAS, in particular KRAS G12C or KRAS G12D oncoprotein. # imgabs0 #

Description

[0001] Citation of Related Applications

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 420,831, filed Oct. 31, 2022, and U.S. Application No. 63 / 510,236, filed Jun. 26, 2023. The entire contents of both applications are incorporated herein by reference. Technical Field

[0003] The present invention relates to inhibitors of the Kirsten rat sarcoma virus (KRAS) oncoprotein, and more particularly to specific 1,5-naphthyridine derivatives, compositions, and methods for treating or preventing diseases, disorders, or medical conditions mediated by KRAS, particularly the KRAS G12C and KRAS G12D oncoproteins. These diseases include various cancers. Background Art

[0004] Ras is a superfamily of small guanosine triphosphate (GTP)-binding proteins consisting of different subtypes. The Ras gene can be mutated into an oncogene associated with many cancers, such as lung cancer, pancreatic cancer, and colon cancer. Ras is one of the most frequently mutated oncogenes. KRAS (Kirsten rat sarcoma virus) is a subtype of Ras and is one of the most frequently mutated Ras genes, accounting for approximately 86% of all known mutations. KRAS plays an on / off switching role in cell signaling. The KRAS protein is a GTPase that operates between an inactive (GDP-bound) and an active (GTP-bound) state to control multiple functions, including cell proliferation. However, mutant KRAS proteins lead to uncontrolled cell proliferation and cancer. The KRAS-4B protein isoform is the major subtype 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). Thus, inhibitors of mutant KRAS proteins that bind to GTP represent potential therapeutic agents for treating various cancers.

[0005] Previous attempts to design inhibitors of the KRAS oncoprotein have mostly been unsuccessful, largely due to the high affinity of the KRAS oncoprotein for GTP. However, recent approaches targeting KRAS G12C have shown greater promise. This mutation is present in approximately 50% of lung cancers and accounts for about 10 - 20% of all KRAS G12 mutations. The mutated cysteine residue is located within the active site, allowing the thiol functional group to form a covalent bond with a suitably functionalized binding ligand (Liu, Acta Pharmaceutica Sinica B 2019). This approach has identified irreversible covalent inhibitors of KRAS G12C that are in clinical studies. The KRAS G12D mutation is present 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, C. L. Cancer Discovery 2017, 7(8), 818 - 831). Given the prominent role of KRAS G12C and KRAS G12D as drivers of many malignancies, there is a need for novel KRAS G12C and KRAS G12D inhibitors with improved selectivity, safety, and efficacy. SUMMARY OF THE INVENTION

[0006] In one aspect, the invention relates to a compound of formula I:

[0007]

[0008] or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein,

[0009] A is selected from optionally 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 、-SR4 、 -S(O)R 4 、 -SO 2 R 4 or one or more substituted aryl or heteroaryl groups in a 5- or 6-membered heterocycle;

[0010] Y and G may be the same or different and are selected from hydrogen, halogen, C 1-4 alkyl, C 1-4 perdeuterated alkyl, -(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);

[0011] L is a bond, O, S or NR 4 ;

[0012] m is 0 - 2;

[0013] n is 0 - 2;

[0014] Z is C(R 4 ) 2 or is selected from C 3-7 cycloalkyl, a saturated or partially unsaturated 4- to 7-membered nitrogen-containing ring, a saturated or partially unsaturated 7- to 10-membered nitrogen-containing bridged bicyclic ring compound;

[0015] R 1 is selected from 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 );

[0016] R 2 is selected from hydrogen, -C(O)CH=CH, -C(O)CF=CH or -C(O)CCl=CH, provided that when R 2 is hydrogen, then m is 1 or 2;

[0017] R 3 is selected from hydrogen, halogen, hydroxy, -C 1-4Alkyl, -C 2-4 Alkenyl, -C 2-4 Alkynyl, -C 0-3 Alkyl(C 3-6 Cycloalkyl), -C 1-4 Alkyl(Halogen), -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 ;

[0018] R 4 is selected from hydrogen, C 1-4 alkyl, aryl or heteroaryl;

[0019] R 5 is selected from hydrogen, hydroxy, C 1-4 alkyl, aryl, heteroaryl, C 1-4 alkoxy, aryloxy or heteroaryloxy;

[0020] R 6 is selected from N(R 4 ) 2 or a 4- to 7-membered saturated or unsaturated heterocycle containing one or more heteroatoms selected from N, O and S.

[0021] On the other hand, the present invention relates to a pharmaceutical composition comprising a compound of formula I or a salt, solvate or prodrug thereof, and a pharmaceutically acceptable carrier.

[0022] On the other hand, the present invention relates to a method of treating a disease, disorder or medical condition in a patient, comprising the step of administering a therapeutic agent to a patient in need thereof, wherein the therapeutic agent is a compound of formula I or a salt, solvate or prodrug thereof. Detailed Description

[0023] The term

[0024] Describe compounds 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 pertains.

[0025] The terms "a" and "an" do not denote a limitation of quantity, but rather denote the presence of at least one of the referenced item. The term "or" means "and / or". The terms "comprising", "having", "including", and "containing" shall be construed as open-ended terms (i.e., meaning "including, but not limited to").

[0026] The recitation of numerical ranges is merely intended as a shorthand method of referring individually to each separate numerical value within the range (unless otherwise indicated herein), and each separate numerical value is incorporated into the specification as if it were recited individually herein. All endpoints of the ranges are included within the range and may be combined independently.

[0027] Unless otherwise indicated herein or clearly contradicted by context, all methods described herein may be performed in a suitable order. The use of any and all examples or exemplary language (e.g., "such as") is merely for the purpose of better illustrating the invention and, unless otherwise claimed, does not limit the scope of the invention. Any language in the specification should not be construed as indicating any unclaimed element as essential to the practice of the invention as used herein. 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 of this disclosure.

[0028] Furthermore, this disclosure encompasses all variations, combinations, and permutations in which one or more limitations, elements, clauses, and descriptive terms from one or more of the listed claims are introduced into another claim. For example, any claim that depends on another claim may be modified to include one or more limitations found in any other claim that depends on the same base claim. In cases where elements are presented as a list, such as in a Markush group format, each subgroup of the elements is also disclosed, and any element may be removed from the group.

[0029] All compounds are understood to include all possible atomic isotopes present in the compound. Isotopes include those atoms having the same atomic number but different mass numbers, and encompass both heavy isotopes and radioactive isotopes. As a general example, but not limited to, isotopes of hydrogen include tritium and deuterium, and isotopes of carbon include 11 C, 13 C, and 14 C. Thus, the compounds disclosed herein may include heavy isotopes or radioactive isotopes in the compound structure, or as substituents attached thereto. Examples of useful heavy isotopes or radioactive isotopes include 18 F,15 N, 18 O, 76 Br, 125 I, and 131 I.

[0030] All formulas disclosed herein include all salts of these formulas.

[0031] The open-ended term "comprising" includes the intermediate term and the closed-ended terms "consisting essentially of" and "consisting of".

[0032] The term "substituted" means that any one or more hydrogens on a specified atom or group are replaced by one selected from the indicated groups, provided that the normal valence of the specified atom is not exceeded. Combinations of substituents and / or variables are permitted only if these combinations result in stable compounds or useful synthetic intermediates. A stable compound or stable structure means a compound that is sufficiently stable to be isolated from a reaction mixture and subsequently formulated into an effective therapeutic agent.

[0033] A dash ("-") that is not between two letters or symbols is used to indicate the point of attachment of a substituent.

[0034] "Alkyl" includes branched and straight-chain saturated aliphatic hydrocarbon groups having a specific number of carbon atoms, typically from 1 to about 8 carbon atoms. The term C 1-6 alkyl, C 1 -C 6 alkyl, and C 1 -C 6 alkyl each denote an alkyl group 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, e.g., C 1-8 alkyl, C 1-4 alkyl, and C 1-2 alkyl. When C 0-n alkyl is used herein in combination with another group, e.g., -C 0-4 alkyl(phenyl), the indicated group (in this case phenyl) is either directly attached by a single covalent bond (C 0 alkyl) or attached by an alkyl chain having a specified number of carbon atoms (in this case 1, 2, 3, or 4 carbon atoms). An alkyl group can also be attached through other groups such as heteroatoms, as in -OC 0-4 alkyl(C 3-7 cycloalkyl). Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, 3-methylbutyl, tert-butyl, n-pentyl, and sec-pentyl.

[0035] "Alkoxy" is an alkyl group as defined above having a specified number of carbon atoms covalently bonded through an oxygen bridge (-O-) to the group to which it is substituted. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, 2-butoxy, tert-butoxy, n-pentyloxy, 2-pentyloxy, 3-pentyloxy, isopentyloxy, neopentyloxy, n-hexyloxy, 2-hexyloxy, 3-hexyloxy, and 3-methylpentyloxy. Similarly, an "alkylthio" or "thioalkyl" group is an alkyl group as defined above having a specified number of carbon atoms covalently bonded through a sulfur bridge (-S-) to the group to which it is substituted. Similarly, "alkenyloxy", "alkynyloxy", and "cycloalkyloxy" refer to alkenyl, alkynyl, and cycloalkyl groups, respectively, each covalently bonded through an oxygen bridge (-O-) to the group to which it is substituted.

[0036] "Halo" or "halogen" refers to fluorine, chlorine, bromine, or iodine and is defined herein to include all of its isotopes, including heavy and radioactive isotopes. Examples of useful halogen isotopes include 18 F, 76 Br, and 131 I. Those skilled in the art will readily appreciate additional isotopes.

[0037] "Haloalkyl" refers to branched and straight-chain alkyl groups having a specific number of carbon atoms that are substituted with one or more halogen atoms, typically up to the maximum allowable number of halogen atoms. Examples of haloalkyl groups include, but are not limited to, trifluoromethyl, difluoromethyl, 2-fluoroethyl, and pentafluoroethyl.

[0038] "Haloalkoxy" is a haloalkyl group as defined above linked through an oxygen bridge (the oxygen of the alcohol group).

[0039] "Peptide" refers to a molecule of a chain of amino acids linked together by an amide bond (also called a peptide bond).

[0040] "Pharmaceutical composition" refers to 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 complies with the GMP (Good Manufacturing Practice) standards of the US FDA for drugs for human or non-human use.

[0041] "Carrier" refers to a diluent, excipient, or solvent administered together with the active compound. "Pharmaceutically acceptable carrier" refers to a substance for preparing a pharmaceutical composition that is generally safe, non-toxic, and has no adverse effects biologically or otherwise, such as an excipient, diluent, or solvent, and includes carriers acceptable for veterinary and human drug use. "Pharmaceutically acceptable carrier" includes one or more such carriers.

[0042] "Patient" refers to 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.

[0043] "Provide" means to give, administer, sell, distribute, transfer (for profit or non-profit), manufacture, formulate or dispense.

[0044] "Treat" or "cure" means to provide an active compound to a patient in an amount sufficient to measurably relieve any symptoms of a disease, slow the progression of the disease or cause the disease to regress. In certain embodiments, treatment of a disease can be initiated before the patient exhibits symptoms of the disease.

[0045] A "therapeutically effective amount" of a pharmaceutical composition is an amount that, when administered to a patient, effectively provides a therapeutic benefit (such as symptom improvement, slowing of disease progression or regression of the disease).

[0046] "Therapeutic compound" refers to a compound that can be used for the diagnosis or treatment of a disease. These compounds can be small molecules, peptides, proteins or other types of molecules.

[0047] A significant change is any detectable change that is statistically significant in a standard parametric test of statistical significance (such as a Student's T-test, where p < 0.05).

[0048] Chemical description

[0049] The compounds of the formulae disclosed herein can contain one or more asymmetric elements, such as stereocenters (e.g., asymmetric carbon atoms), stereogenic axes, rotamers with restricted rotation (e.g., atropisomers), etc., and thus these compounds can exist in different stereoisomeric forms. These compounds can be, for example, racemates or optically active forms. For compounds having two or more asymmetric elements, these compounds can also be mixtures of diastereoisomers. For compounds having an asymmetric center, all optical isomers in pure form and their mixtures are encompassed. In these cases, a single enantiomer, i.e., an optically active form, can be obtained by asymmetric synthesis, synthesis from an optically pure precursor or by resolution of a racemate. Resolution of a racemate can also be accomplished, for example, by conventional methods such as crystallization in the presence of a resolving agent or by chromatography using, for example, a chiral HPLC column. All forms are encompassed herein, regardless of the method used to obtain them.

[0050] All forms of the compounds of the invention (e.g., solvates, optical isomers, enantiomeric forms, polymorphs, prodrugs, free base compounds and salts) can be used alone or in combination.

[0051] The term "chiral" refers to a molecule having the property that its mirror image isomers cannot be superimposed on itself.

[0052] "Stereoisomers" are compounds having the same chemical structure but differing in the arrangement of atoms or groups in space.

[0053] The term "solvate" refers to a chemical complex formed by the interaction of a solvent with a solute, such as a compound of the present invention.

[0054] The term "prodrug" refers to a compound that is inactive biologically but which may be metabolized in vivo to produce a drug.

[0055] "Diastereoisomers" are stereoisomers having two or more chiral centers and whose molecules are not mirror images of one another. Diastereoisomers have different physical properties, such as melting point, boiling point, spectroscopic properties and reactivity. A mixture of diastereoisomers can be separated under high resolution analytical procedures, such as electrophoresis, crystallization in the presence of a resolving agent, or chromatography, using, for example, a chiral HPLC column.

[0056] "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, which may occur in cases where there is no stereoselectivity or stereospecificity in a chemical reaction or process.

[0057] The stereochemical definitions and conventions used herein generally follow S.P. 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. When describing an optically active compound, the prefixes D and L or R and S are used to denote the absolute configuration of the molecule with respect to its chiral center. The prefixes d and l or (+) and (-) are used to denote the sign of rotation of the compound for plane-polarized light, where (-) or l indicates that the compound is levorotatory. A compound prefixed with (+) or d is dextrorotatory.

[0058] A "racemic mixture" or "racemate" is an equimolar (or 50:50) mixture of two enantiomers and has no optical activity. A racemic mixture may occur when there is no stereoselectivity or stereospecificity in a chemical reaction or process.

[0059] "Chelating group" or "chelating agent" is a ligand group that can form two or more independent coordination bonds with a single central atom (usually a metal ion). The chelating groups disclosed herein are organic groups having multiple N, O, or S heteroatoms and having a structure that permits two or more heteroatoms to bond to the same metal ion.

[0060] "Salt" includes derivatives of the disclosed compounds, in which the parent compound is modified by formation of its inorganic and organic acid or base addition salts. The salts of the compounds of the present invention can be synthesized from the parent compounds containing basic or acidic moieties by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid forms of these compounds with a stoichiometric amount of a suitable base (such as hydroxides, carbonates, bicarbonates, etc. of Na, Ca, Mg or K), or by reacting the free base forms of these compounds with a stoichiometric amount of a suitable acid. Such reactions are generally carried out in water or an organic solvent, or in a mixture of both. Generally, when feasible, a non-aqueous medium such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile is used. The salts of the compounds of the present invention further include solvates of the compounds and compound salts. In one embodiment, the compounds of the present invention are synthesized or isolated as trifluoroacetic acid (TFA) salts.

[0061] In one embodiment, the salt forms of the above-described compounds of the present invention may include pharmaceutically acceptable salts. Examples of pharmaceutically acceptable salts include, but are not limited to, non-toxic inorganic or organic acid salts of basic residues (such as amines); alkali metal salts or organic salts of acidic residues (such as carboxylic acids); and the like. Pharmaceutically acceptable salts include, for example, conventional salts and quaternary ammonium salts of the parent compounds formed from non-toxic inorganic or organic acids. 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, nitric acid, etc.); and those formed from organic acids (such as 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, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-aminobenzenesulfonic acid, 2-acetoxybenzoic acid, fumaric acid, p-toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, isethionic acid, HOOC-(CH 2 ) nSalts prepared from -COOH (where n is 0 - 4), etc. A list of other 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.

[0062] In a preferred embodiment, the compounds of formula I are represented by Structures 1a - 1ah and 2a - 2aw shown below, including their pharmaceutically acceptable salts, solvates or prodrugs:

[0063]

[0064]

[0065]

[0066]

[0067]

[0068]

[0069]

[0070]

[0071] The above - mentioned particularly preferred compounds are 1ae, 1af, 1ag, 1ah, 1b, 1d, 1i, 1q, 1s, 1x, 2z, 2al, 2an, 2ap and 2aq.

[0072]

[0073] The compounds disclosed herein can be administered to a patient in the form of a pure chemical substance or a free base chemical substance, but are preferably administered in the form of a pharmaceutical composition. Accordingly, the present invention encompasses pharmaceutical compositions that include a compound or a salt of the compound (including pharmaceutically acceptable salts), such as a compound of Formula I, and at least one pharmaceutically acceptable carrier. The pharmaceutical composition may contain the compound or salt of Formula I as the sole active agent, but preferably contains at least one additional active agent. In certain embodiments, the dosage form of the pharmaceutical composition contains from about 0.1 mg to about 2000 mg, from about 10 mg to about 1000 mg, from about 100 mg to about 800 mg, or from about 200 mg to about 600 mg of the compound of Formula I, and optionally from about 0.1 mg to about 2000 mg, from about 10 mg to about 1000 mg, from about 100 mg to about 800 mg, or from about 200 mg to about 600 mg of an additional active agent in a unit dosage form. The pharmaceutical composition may also include a certain molar ratio of the compound (such as a compound of Formula I) to an additional active agent. For example, the pharmaceutical composition may contain an additional active agent and a compound of Formula I in 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. Particularly preferred forms of Formula I for use in pharmaceutical compositions include compound 1ae-1ah, 1b, 1d, 1q, 2z, or 2al, or a salt, solvate, or prodrug thereof, and a pharmaceutically acceptable carrier.

[0074] The compounds disclosed herein can be administered orally, topically, parenterally, by inhalation or spray, sublingually, transdermally, buccally, rectally, as an ophthalmic solution, or by other means, in dosage unit formulations containing conventional pharmaceutically acceptable carriers. The pharmaceutical composition can be formulated in any pharmaceutically useful form, such as an aerosol, cream, gel, pill, capsule, tablet, syrup, transdermal patch, or ophthalmic solution. Some dosage forms, such as tablets and capsules, are subdivided into unit doses of appropriate size containing an appropriate amount (e.g., an effective amount for the desired purpose) of the active ingredient.

[0075] The carrier includes excipients and diluents and must have a sufficiently high purity and a sufficiently low toxicity to render them suitable for administration to the patient being treated. The carrier can be inert, or it can have a pharmaceutical benefit of its own. The amount of carrier used in combination with the compound is sufficient to provide a practical amount of the administered substance per unit dose of the compound.

[0076] The classes of carriers include, but are not limited to, binders, buffers, colorants, diluents, disintegrants, emulsifiers, flavoring agents, glidants, lubricants, preservatives, stabilizers, surfactants, tablet binders, and wetting agents. Some carriers may be included in more than one class; for example, vegetable oil may be used as a lubricant in some formulations and as a diluent in other formulations. Exemplary pharmaceutically acceptable carriers include sugars, starches, celluloses, tragacanth powder, malt, gelatin, talc, and vegetable oils. The pharmaceutical composition may optionally contain an active agent that does not substantially interfere with the activity of the compounds of the present invention.

[0077] The pharmaceutical composition / combination can be formulated for oral administration. These compositions contain from 0.1 to 99 weight percent (wt%) of the compound of formula I, and typically contain at least about 5 wt% of the compound of formula I. Some embodiments contain from about 25 wt% to about 50 wt% or from about 5 wt% to about 75 wt% of the compound of formula I.

[0078] Methods of treatment

[0079] The compound of formula I and the pharmaceutical composition containing the same can be used for diagnosing or treating diseases, disorders, or medical conditions mediated by KRAS, especially KRAS mutants G12C and G12D, including various cancers, such as glioma (glioblastoma), acute myeloid leukemia, acute myelocytic leukemia, myelodysplastic / myeloproliferative neoplasms, sarcoma, chronic myelomonocytic leukemia, non-Hodgkin lymphoma, astrocytoma, melanoma, non-small cell lung cancer, cholangiocarcinoma, chondrosarcoma, colon cancer, or pancreatic cancer.

[0080] According to the present invention, a method for a KRAS-mediated disease or condition includes providing a therapeutically effective amount of the compound of formula I to a patient in need thereof. In one embodiment, the patient is a mammal, more specifically a human. As will be understood by those skilled in the art, the present invention also encompasses methods for treating non-human patients (such as companion animals, e.g., cats, dogs, and livestock).

[0081] The therapeutically effective amount of the pharmaceutical composition is preferably an amount sufficient to alleviate or improve the symptoms of the disease or condition. For example, in the case of a KRAS-mediated disease, the therapeutically effective amount may be an amount sufficient to alleviate or improve cancer. When administered to a patient, the therapeutically effective amount of the compound or pharmaceutical composition described herein will also provide a sufficient concentration of the compound of formula I. The sufficient concentration is preferably the concentration of the compound required in the patient's body to prevent or counteract the condition. Such an amount can be determined experimentally, for example, by measuring the blood concentration of the compound, or theoretically by calculating the bioavailability.

[0082] According to the present invention, the treatment methods disclosed herein include providing a patient with a dose of a compound of Formula I. A dosage level of from about 0.1 mg to about 140 mg per kilogram of body weight per day of each compound can be used to treat the above-mentioned conditions (from about 0.5 mg to about 7 g per patient per day). The amount of the compound that can be combined with a carrier material to produce a single dosage form will vary depending on the patient being treated and the particular mode of administration. The unit dosage form generally contains from about 1 mg to about 500 mg of each active compound. In certain embodiments, a patient is provided with from 25 mg to 500 mg, or from 25 mg to 200 mg, of the compound of Formula I per day. 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 conditions, a dosing regimen of 4 times a day or less can be used, and in certain embodiments, a dosing regimen of once or twice a day is used.

[0083] However, it should be understood that the specific dosage 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, diet, time of administration, route of administration, rate of excretion, drug combination, and the severity of the particular disease being treated.

[0084] The compounds of Formula I can be administered alone (i.e., the sole therapeutic agent in a regimen) to treat or prevent KRAS-mediated diseases and conditions, such as various cancers, or can be administered in combination with another active agent. One or more compounds of Formula I can be administered in combination with a regimen of one or more other active agents (such as anticancer cytotoxic agents). In one embodiment, a method of 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.

[0085] As will be understood by those skilled in the art, the treatment methods provided herein can also be used to treat mammals other than humans, including for veterinary applications, such as treating horses and domestic animals, such as cattle, sheep, cows, goats, pigs, etc., and pets (companion animals), such as dogs and cats.

[0086] For diagnostic or research applications, a variety of mammals will be suitable subjects, including rodents (e.g., mice, rats, hamsters), rabbits, primates, and pigs (such as inbred pigs), etc. In addition, for in vitro applications, such as in vitro diagnostic and research applications, body fluids (e.g., blood, plasma, serum, interstitial fluid, saliva, feces, and urine) as well as cell and tissue samples from the above-mentioned subjects will be applicable.

[0087] In one embodiment, the present invention provides a method for treating a disease, disorder or medical condition mediated by KRAS (especially the KRAS mutant G12C), including various cancers, for a patient determined to be 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.

[0088] In another embodiment, a method for treating or diagnosing a KRAS-mediated disease or condition may further comprise co-administering a compound of formula I with one or more additional compounds to a patient in need of such treatment, wherein at least one of the additional compounds is an active agent. The one or more additional compounds may include additional therapeutic compounds, including anti-cancer therapeutic compounds, such as doxorubicin, paclitaxel, docetaxel, cisplatin, camptothecin, temozolomide, bevacizumab, trastuzumab, cetuximab, EGFR inhibitors, osimertinib, razertinib, CDK4 / 6 inhibitors, abemaciclib, palbociclib, ribociclib, c-MET inhibitors, capmatinib, volitinib, ALK inhibitors, crizotinib, alectinib, ceritinib, brigatinib, entrectinib, lorlatinib, PD-1 antagonists, PD-L1 antagonists, ipilimumab, pembrolizumab, nivolumab, etc.

[0089] Examples

[0090] Chemical synthesis

[0091] The compounds of formula 1 described herein and / or their pharmaceutically acceptable salts can be synthesized from commercially available starting materials by methods well known to synthetic organic chemists in the art. The following general synthetic schemes 1 and 2 illustrate representative methods for preparing the compounds of most of the examples. In certain embodiments, when the Suzuki cross-coupling reaction of arylboronic acid / boronate with organic halide / pseudohalide (Beketskaya, I.P. et al., Coordin. Chem. Rev. 2019, 385, 137-173) is impractical or unsuccessful, then the Stille cross-coupling reaction of organotin with organic halide / pseudohalide can be used as an alternative method (Espinet, P. et al., ACS Catal. 2015, 5, 3040-3053). Many of the required intermediates can be prepared as described in WO2021041671. The listed starting materials, reactions, reagents, solvents, temperatures, catalysts and ligands are not limited to those described for illustrative purposes only. For clarity, some abbreviations and acronyms well known to those skilled in the art that may be used in Schemes 1 and 2 and the examples are listed below.

[0092] The synthesis of the compounds of the invention is illustrated by the sequence of steps shown in Scheme 1. In Scheme 1, in a reaction such as CH 2 Cl 2 The 1,5-naphthyridine derivative 3 was oxidized with mCPBA in a solvent to generate the N-oxide compound 4. At elevated temperature, 4 reacted with POCl 3 The reaction afforded the corresponding chloro derivative 5. 5 was reacted with 6 to give compound 7. The reaction was carried out using a suitable base (such as sodium hydride, Hünig's base, K 2 CO 3 or Cs 2 CO 3 8 can be treated with 7 in a polar aprotic solvent such as N-methyl-2-pyrrolidone at room temperature (RT) or elevated temperature to give compound 9. Suzuki-Miyaura coupling of 9 with a boronic ester such as 10 (or the corresponding boronic acid) under standard conditions in a mixed solvent such as 1,4-dioxane and water can be used to prepare 11. Under acidic conditions (such as anhydrous 4M HCl in 1,4-dioxane or in CH 2 Cl 2 The Boc protecting group of 11 was removed by TFA in the reaction mixture to generate R 2 In a dichloromethane solvent containing a base (such as triethylamine), 12 is acylated with acryloyl chloride 13 to generate R 2 The corresponding compound 14 is C(O)CH=CH, -C(O)CF=CH or -C(O)CCl=CH.

[0093] Solution 1

[0094]

[0095] Abbreviations and acronyms

[0096] The following abbreviations and acronyms may be used in this application:

[0097] anhyd.=without water;

[0098] aq.=aqueous solution;

[0099] B 2 pin 2 =Bis(pinacol)diboron;

[0100] Boc = tert-butyloxycarbonyl;

[0101] n-Bu 3 P = tri-tert-butylphosphine;

[0102] Compd = Compound;

[0103] d = days;

[0104] DCM = Dichloromethane;

[0105] DIEA = DIPEA = N,N - Diisopropylethylamine;

[0106] DMF = N,N - Dimethylformamide;

[0107] DMSO = Dimethyl sulfoxide;

[0108] DMA = N,N - Dimethylacetamide;

[0109] dppf = 1,1′ - Bis(diphenylphosphino)ferrocene;

[0110] DTBPF = 1,1′ - Bis(di - tert - butylphosphino)ferrocene;

[0111] EtOAc = Ethyl acetate;

[0112] equiv = equivalent;

[0113] Ex = Example;

[0114] h = hours;

[0115] KOAc = Potassium acetate;

[0116] LiHMDS = Lithium bis(trimethylsilyl)amide [LiN(SiMe 3 ) 2 ;

[0117] mCPBA = meta - Chloroperoxybenzoic acid;

[0118] MeOH = Methanol;

[0119] NMP = N - Methyl - 2 - pyrrolidone;

[0120] min = minutes;

[0121] Pd(dppf)Cl 2 = [1,1′ - Bis(diphenylphosphino)ferrocene]dichloropalladium(II);

[0122] RT = Room temperature;

[0123] satd. = Saturated solution;

[0124] TEA = Triethylamine;

[0125] TFA = Trifluoroacetic acid;

[0126] THF = Tetrahydrofuran;

[0127] The concepts of the present invention have been described in accordance with exemplary principles and embodiments. However, those skilled in the art will recognize that changes and equivalent substitutions can be made to the described content without departing from the scope and spirit of the present disclosure as defined by the following claims.

[0128] Example 1

[0129] 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).

[0130]

[0131] Example 1 (1b) was prepared as shown in Scheme 2 below.

[0132] Scheme 2

[0133]

[0134] 7 - Bromo - 4 - chloro - 1,5 - naphthyridin - 1 - oxide (16). This compound was prepared as described on pages 64 - 65 of WO2020150114 by reacting mCPBA with 3 - bromo - 8 - chloro - 1,5 - naphthyridine (15; CAS#97267 - 61 - 3; 1.70 g, 7.02 mmol) in CH 2 Cl 2 to give 1.50 g (83%) of 7 - bromo - 4 - chloro - 1,5 - naphthyridin - 1 - oxide (16) as a pale yellow solid: HPLC - MS (ES + ) m / z [M + H + = 259, 261, 263; 1 1H NMR (300 MHz, CDCl 3 ) δ 9.24 (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).

[0135] 7 - Bromo - 2,4 - dichloro - 1,5 - naphthyridine (17). This compound was prepared as described on page 65 of WO2020150114 from 7 - bromo - 4 - chloro - 1,5 - naphthyridin - 1 - oxide (16; 775 mg, 3.00 mmol) to give 750 mg (90%) of 7 - bromo - 2,4 - dichloro - 1,5 - naphthyridine (17) as a pink solid: HPLC - MS (ES + ) m / z [M + H+ = 277, 279, 281, 283, 1 H NMR (300 MHz, CDCl 3 ) δ 9.05 (d, J = 2.1 Hz, 1H), 8.51 (d, J = 2.1 Hz, 1H), 7.78 (s, 1H).

[0136] 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 the mixture was stirred at room temperature. 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 addition, the ice bath was removed, and the reaction mixture was heated to reflux for 23 h. The mixture was cooled to room temperature, 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 (aq.) (3X), dried (MgSO 4 ), filtered, and concentrated in vacuo. The crude product was purified by silica gel column chromatography, eluting with a gradient of 10% to 40% EtOAc in hexane, 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, CDCl 3 ) δ 8.89 (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).

[0137] 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 Cs 2 CO 3 (545 mg, 1.68 mmol) in CH 3 CN (8 mL) was heated to reflux. After 72 h, the mixture was cooled to room temperature, diluted with EtOAc, washed with saturated aqueous NaCl solution (3X), dried (MgSO 4 ), filtered, and concentrated in vacuo. 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; 1 H NMR (300 MHz, CDCl 3 ) δ 8.72 (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).

[0138] 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). 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 K 2 CO 3 (487 mg, 3.53 mmol) in a mixture of 1,4-dioxane (3 mL) and water (1.4 mL) was degassed by bubbling with N 2 for 30 minutes with stirring.

[0139] Tetrakis(triphenylphosphine)palladium(0) (33 mg, 0.029 mmol) was added and the reaction mixture was degassed by bubbling with N 2 for a further 20 minutes with stirring. The reaction mixture was stirred and heated at 80 °C for 16 h under an N 2 atmosphere. The reaction mixture was cooled to room temperature, diluted with EtOAc, filtered through celite, and then the filtrate was washed with saturated aqueous NaCl solution (3X), dried (MgSO 4 ), filtered and concentrated in vacuo. The crude product was purified by silica gel column chromatography, eluting with a gradient of 0% to 10% MeOH in DCM to give 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; 1 H NMR (300 MHz, CDCl 3)δ 8.67 (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).

[0140] 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). 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) was dissolved in CH 2 Cl 2 (2 mL), and 4 M HCl / 1,4 - dioxane solution (0.2 mL) was added to this solution, and the mixture was stirred at room temperature. After 16 h, 0.1 M NaOH solution was added to the mixture until the pH was basic. The layers were separated, and the aqueous layer was extracted with CH 2 Cl 2 (3X), dried (MgSO 4 ), filtered, and concentrated in vacuo 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 1H NMR (300 MHz, CDCl 3)δ 8.64 (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).

[0141] 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). 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) was dissolved in CH 2 Cl 2 (8 mL), and Et 3 N (16 μL, 0.114 mmol) was added to this solution, and the mixture was stirred at room temperature. The mixture was cooled to 0 °C, and acryloyl chloride (25; CAS# 814 - 68 - 6; 10 μL, 0.114 mmol) was added. After 1 h at 0 °C, the mixture was diluted with CH 2 Cl 2 , and washed with H 2 O (2X). The layers were separated, dried (MgSO 4 ), and the CH 2 Cl 2 layer was filtered and concentrated in vacuo. The crude product was purified by silica gel column chromatography, eluting with a 0% to 10% gradient of MeOH in DCM containing 5% NH 4 OH (v / v), to afford 20 g (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; 11H NMR (300 MHz, CDCl 3 ) δ 8.69 (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).

[0142] Example 2

[0143] 2 - ((S)-4-(7-(8 - chloronaphthalen - 1 - yl)-2 - (((S)-1 - methylpyrrolidin - 2 - yl)methoxy)-1,5 - naphthyridin - 4 - yl)-1-(2 - fluoroprop - 2 - enoyl)piperazin - 2 - yl)acetonitrile (1q).

[0144]

[0145] Example 2 (1q) was prepared as shown in Scheme 3 below.

[0146] Scheme 3

[0147]

[0148] 2 - ((S)-4-(7-(8 - chloronaphthalen - 1 - yl)-2 - (((S)-1 - methylpyrrolidin - 2 - yl)methoxy)-1,5 - naphthyridin - 4 - yl)-1-(2 - fluoroprop - 2 - enoyl)piperazin - 2 - yl)acetonitrile (1q). Under N 2 atmosphere, 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 EtOAc (4 mL) was added propylphosphonic anhydride solution (T 3P (0.2 mL, 0.275 mmol, 50% EtOAc) was added and stirred at room temperature. After 40 minutes, the reaction mixture was diluted with EtOAc and washed with saturated NaHCO 3 aqueous solution (3X) and saturated NaCl aqueous solution (2X). Subsequently, the organic layer was dried (MgSO 4 ), filtered, and concentrated in vacuo. The resulting crude product was purified by silica gel column chromatography, eluting with a gradient of 10% to 100% EtOAc containing 1% Et 3 N (v / v) in DCM to give 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-fluoroprop-2-enoyl)piperazin-2-yl)acetonitrile (1q) as an off-white solid: HPLC-MS (ES + ) m / z [M+H + = 599, 601; 1 H NMR (300 MHz, CDCl 3 ) δ 8.69 (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).

[0149] Example 3

[0150] 1-(8-(8-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-6-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-1,5-naphthyridin-3-yl)-2-fluoro-6-hydroxynaphthalen-1-yl)ethan-1-one (2aw).

[0151]

[0152] Example 3 (2aw) was prepared as shown in Scheme 4 below.

[0153] Scheme 4

[0154]

[0155] 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 to 90 °C under N 2 atmosphere. After 16 h, the mixture was cooled to room temperature, diluted with EtOAc, and the filtrate was washed with saturated aqueous NaCl solution (3X), dried (MgSO 4 ), filtered, and concentrated in vacuo. 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 1H NMR (300 MHz, CDCl 3 ) δ 8.75 (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).

[0156] tert-Butyl (1R,5S)-3-(7-bromo-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (30). Under N 2Under an atmosphere, at 0 °C cooling, sodium hydride (38 mg, 0.953 mmol, 60% mineral oil dispersion) was added to a solution of (2R,7aS)-2-fluoro-1,2,3,7a-tetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol (29; CAS#2097518-76-6; 151 mg, 0.953 mmol) in anhydrous DMF (6 mL). After 35 minutes, 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 warmed to room temperature. After 16 h, the mixture was diluted with EtOAc, and the filtrate was washed with saturated aqueous NaCl solution (4X), dried (MgSO 4 ), filtered and concentrated in vacuo. The crude product was purified by silica gel column chromatography, eluting with a gradient of 0% to 10% MeOH in DCM, to afford 170 mg (46%) of tert-butyl (1R,5S)-3-(7-bromo-2-(((2R,7aS)-2-fluoro-1,2,3,7a-tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (30) as a white foam: HPLC-MS (ES + ) m / z [M+H + = 576, 578; 1 H NMR (300 MHz, CDCl 3 ) δ 8.60 (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).

[0157] tert-Butyl (1R,5S)-3-(7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)-naphthalen-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-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-pyrrolizin-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 K 2 CO 3 (156 mg, 1.14 mmol) were mixed in 1,4-dioxane (3 mL) and water (0.5 mL), and the mixture was degassed by bubbling with N 2 for 30 minutes. Tetrakis(triphenylphosphine)palladium(0) (33.3 mg, 0.028 mmol) was added, and the reaction mixture was degassed by bubbling with N 2 and stirred for an additional 20 minutes. The reaction mixture was stirred and heated at 80 °C for 16 h under an N 2 atmosphere, cooled to room temperature, diluted with EtOAc and filtered through celite. The filtrate was washed with saturated aqueous NaCl solution (2X), dried (MgSO 4 ), filtered and concentrated in vacuo. The crude product was a mixture of 32 and 33, which was purified by silica gel column chromatography, eluting with a gradient of 30% to 100% EtOAc in hexanes, to afford 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-pyrrolizin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (32) as an off-white solid, a mixture of atropisomers (HPLC-MS (ES+) m / z [M+H+] = 882), and 67 mg (33%) of 33 as an orange-yellow foamy solid (HPLC-MS (ES+) m / z [M+H+] = 726).

[0158] tert-Butyl (1R,5S)-3-(7-(8-ethynyl-7-fluoro-3-(methoxymethoxy)naphthalen-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-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 solution in THF) was added to a solution of tert-butyl (1R,5S)-3-(7-(7-fluoro-3-(methoxymethoxy)-8-((triisopropylsilyl)ethynyl)naphthalen-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-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 room temperature. After 1 h, the mixture was diluted with EtOAc, washed with saturated aqueous NaCl (3X), dried (MgSO 4 ), filtered, and concentrated in vacuo 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-pyrrolizin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (33) as an orange-yellow foamy solid: HPLC-MS (ES + ) m / z [M+H + = 726; 1 H NMR (300 MHz, CDCl 3 ) δ 9.04 (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).

[0159] 1-(8-(8-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-6-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-1,5-naphthyridin-3-yl)-2-fluoro-6-hydroxynaphthalen-1-yl)ethan-1-one (2aw). Under N 2 atmosphere, 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-pyrrolizin-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 room temperature. After 1 h, the mixture was diluted with DCM and slowly transferred by pipette to an aqueous solution of NH 4 OH (25 mL), and stirred at room temperature. The phases were separated, the aqueous layer was extracted with EtOAc (2X), and the combined organic layers were dried (MgSO 4 ), filtered, and concentrated in vacuo. The crude product was purified by silica gel column chromatography, eluting with a 0% to 20% gradient of MeOH in DCM containing 5% NH 4 OH (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-pyrrolizin-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; 1 H NMR (300 MHz, DMSO-d 6)δ 10.92 (broad singlet, 1H), 10.26 (singlet, 1H), 9.29 (broad singlet, 1H), 9.11 (broad singlet, 1H), 8.62 (doublet, J = 2.0 Hz, 1H), 8.02 (doublet of doublets, J = 4.6, 6.0 Hz, 1H), 7.91 (doublet, J = 2.0 Hz, 1H), 7.46 (triplet, J = 9.3 Hz, 1H), 7.40 (doublet, J = 2.3 Hz, 1H), 7.13 (doublet, J = 2.0 Hz, 1H), 6.62 (singlet, 1H), 5.59 (doublet, J = 52.4 Hz, 1H), 4.71 - 4.45 (multiplet, 3H), 4.36 (doublet, J = 12.3 Hz, 1H), 4.25 (broad singlet, 2H), 4.07 - 3.65 (multiplet, 4H), 2.37 - 2.01 (multiplet, 10H), 1.99 (singlet, 3H).

[0160] Example 4

[0161] 4-(8-((1R,5S)-3,8-Diazabicyclo[3.2.1]octan-3-yl)-6-((tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-1,5-naphthyridin-3-yl)naphthalen-2-ol (2a).

[0162]

[0163] Example 4 (2a) was prepared as shown in Scheme 5 below.

[0164] Scheme 5

[0165]

[0166] tert-Butyl-(1R,5S)-3-(7-bromo-2-((tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (35). Under N 2 atmosphere, sodium hydride (70 mg, 1.72 mmol, 60% w / w mineral oil dispersion) was added to a solution of hexahydro-1H-pyrrolizin-7a-ylmethanol (34; CAS# 78449-72-6; 242 mg, 1.72 mmol) in anhydrous THF (8 mL) at 0 °C. After 30 minutes, 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 room temperature, diluted with EtOAc, and the filtrate was washed with saturated aqueous NaCl solution (3X) and dried (MgSO4 ) and filtered and concentrated in vacuo. The crude product was purified by silica gel column chromatography, eluting with a gradient of 0% to 10% MeOH in DCM containing 10% NH 4 OH (v / v) to afford 300 mg (47%) of tert-butyl-(1R,5S)-3-(7-bromo-2-((tetrahydro-1H-pyrrolizin-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, CDCl 3 ) δ 8.59 (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).

[0167] tert-Butyl (1R,5S)-3-(7-(3-hydroxynaphthalen-1-yl)-2-((tetrahydro-1H-pyrrolizin-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-pyrrolizin-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), K 2 CO 3 (293 mg, 2.13 mmol) in 1,4-dioxane (5 mL) and water (1 mL) was degassed by bubbling N 2 for 30 minutes. Tetrakis(triphenylphosphine)palladium(0) (60 mg, 0.052 mmol) was added and the reaction mixture was further degassed by bubbling N 2 for 20 minutes. After degassing was complete, the reaction mixture was placed under N 2Under the atmosphere, stir and heat at 85 °C for 16 h. Cool the reaction mixture to room temperature, dilute with EtOAc, and filter through diatomaceous earth. Wash the organic layer with saturated aqueous NaCl solution (3X), dry (MgSO 4 ), filter, and concentrate in vacuo. Purify the crude product by silica gel column chromatography, eluting with a gradient of 0% to 10% MeOH in DCM containing 10% NH 4 OH (v / v) to afford 184 mg (57%) of tert-butyl (1R,5S)-3-(7-(3-hydroxynaphthalen-1-yl)-2-((tetrahydro-1H-pyrrolizin-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; 1 H NMR (300 MHz, DMSO-d 6 ) δ 9.98 (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).

[0168] 4-(8-((1R,5S)-3,8-Diazabicyclo[3.2.1]octan-3-yl)-6-((tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-1,5-naphthyridin-3-yl)naphthalen-2-ol (2a). To a solution of tert-butyl (1R,5S)-3-(7-(3-hydroxynaphthalen-1-yl)-2-((tetrahydro-1H-pyrrolizin-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 added a solution of 4 M HCl in 1,4-dioxane (3 mL) under N 2An orange suspension was obtained by stirring at room temperature under the atmosphere. After 2 h, the reaction was allowed to stand at room temperature. After 72 h, MeOH was added to the mixture, diluted with DCM, and then the resulting solution was slowly transferred by pipette to NH 4 OH(aq), and stirred at room temperature. Phase separation occurred, and the aqueous layer was extracted once with EtOAc and then once with DCM. The organic layers were combined, dried (MgSO 4 ), filtered and concentrated in vacuo. The resulting crude product was purified by silica gel column chromatography, eluting with a 0% to 15% gradient of MeOH containing 10% NH 4 OH (v / v) in DCM to give 102 mg (71%) of 4-(8-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-6-((tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-1,5-naphthyridin-3-yl)naphthalen-2-ol (2a) as a brown solid: HPLC-MS (ES + ) m / z [M+H + = 522; 1 H NMR (300 MHz, DMSO-d 6 ) δ 10.0 (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).

[0169] Example 5

[0170] (S)-2-(1-acryloyl-4-(7-(8-chloronaphthalen-1-yl)-2-((tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)piperazin-2-yl)acetonitrile (1ae).

[0171]

[0172] Example 5 (1ae) was prepared as shown in Scheme 6 below.

[0173] Scheme 6

[0174]

[0175] tert-Butyl (S)-4-(7-bromo-2-((tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (38). Under N 2 atmosphere, sodium hydride (142 mg, 3.54 mmol, 60% w / w mineral oil dispersion) was added to a solution of hexahydro-1H-pyrrolizin-7a-ylmethanol (34; CAS# 78449-72-6; 500 mg, 3.54 mmol) in anhydrous THF (20 mL) at 0 °C. After 30 minutes, 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 room temperature, diluted with EtOAc, washed with saturated aqueous NaCl solution (3X), dried (MgSO 4 ) and filtered, and concentrated in vacuo. The crude product was purified by silica gel column chromatography, eluting with a gradient of 0% to 6% MeOH in DCM containing 10% NH 4 OH (v / v), to give 734 mg (54%) of tert-butyl (S)-4-(7-bromo-2-((tetrahydro-1H-pyrrolizin-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; 1 1H NMR (300 MHz, CDCl 3 ) δ 8.71 (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).

[0176] tert-Butyl (S)-4-(7-(8-chloronaphthalen-1-yl)-2-((tetrahydro-1H-pyrrolizin-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-pyrrolizin-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 K 2 CO 3 (718 mg, 5.21 mmol) in dioxane (12 mL) and water (2.4 mL) was degassed by bubbling with N 2 for 30 minutes with stirring. Tetrakis(triphenylphosphine)palladium(0) (147 mg, 0.127 mmol) was added and the reaction mixture was degassed by continuing to bubble with N 2 for 20 minutes with stirring. The reaction mixture was stirred and heated at 80 °C for 16 h under an N 2 atmosphere. The reaction mixture was cooled to room temperature, diluted with EtOAc and filtered through celite. Washed with saturated aqueous NaCl solution (3X), dried (MgSO 4 ), filtered and concentrated in vacuo. The crude product was purified by silica gel column chromatography (eluting with a gradient of 0% to 6% MeOH in DCM containing 10% NH 4 OH (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; 1 H NMR (300 MHz, CDCl 3)δ 8.66 (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).

[0177] (S)-2-(4-(7-(8-chloronaphthalen-1-yl)-2-((tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)piperazin-2-yl)acetonitrile (40). To 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 CH 2 Cl 2 (10 mL) was slowly added dropwise 4 M Hcl / dioxane solution (3.3 mL), thereby forming an orange viscous solid, and the mixture was stirred at room temperature. After 3 h, the solid was sampled and determined to be the product by LC / MS. The DCM layer was decanted off, and then the viscous solid was dissolved in MeOH. The solution was diluted with saturated NaHCO 3 aqueous solution, extracted with DCM (5X), dried (MgSO 4 ), filtered, and concentrated in vacuo. The crude product was purified by silica gel column chromatography, eluting with a 0% to 10% gradient of MeOH in DCM containing 10% NH 4 OH (v / v), to give 191 mg (65%) of (S)-2-(4-(7-(8-chloronaphthalen-1-yl)-2-((tetrahydro-1H-pyrrolizin-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, CDCl 3)δ8.63(d, J = 2.1Hz, 1H), 8.00(d, J = 2.0Hz, 1H), 7.96(d, J = 8.1Hz, 1H), 7.88(d, J = 8.1Hz, 1H), 7.60 - 7.49(m, 2H), 7.47 - 7.36(m, 2H), 6.45(s, 1H), 4.23(d, J = 4.8Hz, 2H), 3.95(dd, J = 11.4, 33.2Hz, 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).

[0178] (S)-2-(1 - acryloyl - 4-(7-(8 - chloronaphthalen - 1 - yl)-2 - ((tetrahydro - 1H - pyrrolizin - 7a(5H)-yl)methoxy)-1,5 - naphthyridin - 4 - yl)piperazin - 2 - yl)acetonitrile (1ae). To a solution of (S)-2-(4-(7-(8 - chloronaphthalen - 1 - yl)-2 - ((tetrahydro - 1H - pyrrolizin - 7a(5H)-yl)methoxy)-1,5 - naphthyridin - 4 - yl)piperazin - 2 - yl)acetonitrile (40; 83 mg, 0.15 mmol) in CH 2 Cl 2 (12 mL) was added Et 3 N (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 diluted with CH 2 Cl 2 , washed with H 2 O (2X), dried (MgSO 4 ), filtered, and concentrated in vacuo. The crude product was purified by silica gel column chromatography (eluted with a 0% to 5% gradient of MeOH in DCM containing 10% NH 4 OH (v / v)), to give 45 mg (49%) of (S)-2-(1 - acryloyl - 4-(7-(8 - chloronaphthalen - 1 - yl)-2 - ((tetrahydro - 1H - pyrrolizin - 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 HNMR (300 MHz, CDCl 3)δ 8.68 (broad 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 (broad 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).

[0179] Example 6

[0180] (S)-2-(4-(7-(8-Chloronaphthalen-1-yl)-2-((tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-1-(2-fluoroprop-2-enoyl)piperazin-2-yl)acetonitrile (1af).

[0181]

[0182] Example 6 (1af) was prepared as shown in Scheme 7 below.

[0183] Scheme 7

[0184]

[0185] (S)-2-(4-(7-(8-Chloronaphthalen-1-yl)-2-((tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-1-(2-fluoroprop-2-enoyl)piperazin-2-yl)acetonitrile (1af). Under N 2 atmosphere, propylphosphonic anhydride solution (T 3 P, 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-pyrrolizin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)piperazin-2-yl)acetonitrile (40; 83 mg, 0.15 mmol), 2-fluoro-2-propenoic acid (26; CAS# 430 - 99 - 9; 41 mg, 0.45 mmol) and diisopropylethylamine (0.27 mL, 1.50 mmol) in EtOAc (11 mL), and the mixture was stirred at room temperature. After 40 minutes, an additional propylphosphonic anhydride solution (T 3P, 0.3 mL), and the mixture was stirred at room temperature. After 1.5 h, the reaction mixture was diluted with EtOAc and washed with 5% aqueous NaHCO 3 solution (3X), dried (MgSO 4 ), filtered, and concentrated in vacuo. The crude product was purified by silica gel column chromatography (eluted with a gradient of 10% to 100% EtOAc containing 5% Et 3 N (v / v)) to give 25 mg (27%) of (S)-2-(4-(7-(8-chloronaphthalen-1-yl)-2-((tetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-1-(2-fluoropropionyl)piperazin-2-yl)acetonitrile (1af) as an off-white powder: HPLC-MS (ES + ) m / z [M+H + = 625, 627; 1 H NMR (300 MHz, CDCl 3 ) δ 8.68 (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 (brd, 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).

[0186] Example 7

[0187] 2-((S)-1-Propionyl-4-(7-(8-chloronaphthalen-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)piperazin-2-yl)acetonitrile (1ag).

[0188]

[0189] Example 7 (1ag) was prepared as shown in Scheme 8 below.

[0190] Scheme 8

[0191]

[0192] tert-Butyl (S)-4-(7-bromo-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (41). Under N 2 atmosphere, sodium hydride (142 mg, 3.54 mmol, 60% w / w mineral oil dispersion) was added to a solution of (2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol (29; CAS# 2097518-76-6; 563 mg, 3.54 mmol) in anhydrous THF (20 mL) at 0 °C. After 30 minutes, 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 room temperature. After 30 minutes, the mixture was heated to reflux. After 16 h, the mixture was cooled to room temperature, diluted with EtOAc, washed with saturated aqueous NaCl (aq.) (3X), dried (MgSO 4 ), filtered, and concentrated in vacuo. The crude solid was suspended in Et 2 O and stirred at room temperature. After 16 h, the solid was filtered, washed with Et 2 O, and dried to give 900 mg (65%) of tert-Butyl (S)-4-(7-bromo-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-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; 1 H NMR (300 MHz, CDCl 3 ) δ 8.72 (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).

[0193] tert-Butyl (S)-4-(7-(8-chloronaphthalen-1-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (42). tert-Butyl (S)-4-(7-bromo-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-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 K 2 CO 3 (563 mg, 4.08 mmol) in a mixture of dioxane (7 mL) and water (4 mL) was degassed by bubbling with N 2 for 20 minutes with stirring. Tetrakis(triphenylphosphine)palladium(0) (78 mg, 0.068 mmol) was added, and the reaction mixture was degassed by continuing to bubble with N 2 for 20 minutes with stirring. The reaction mixture was stirred and heated at 80 °C for 16 h under a N 2 atmosphere. The reaction mixture was cooled to room temperature and diluted with EtOAc. Washed with saturated aqueous NaCl solution (3X), dried (MgSO 4 ), filtered, and concentrated in vacuo. 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-pyrrolizin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-2-(cyanomethyl)piperazine-1-carboxylate (42) as an off-white solid: HPLC-MS (ES + ) m / z [M + H + = 671, 673; 1 H NMR (300 MHz, CDCl 3)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).

[0194] 2 - ((S)-4-(7-(8 - chloronaphthalen - 1 - yl)-2 - (((2R,7aS)-2 - fluorotetrahydro - 1H - pyrrolizin - 7a(5H)-yl)methoxy)-1,5 - naphthyridin - 4 - yl)piperazin - 2 - yl)acetonitrile (43). Under N 2 atmosphere, 4M HCl / dioxane (3.6 mL) solution was slowly added dropwise to a solution of tert - butyl (S)-4-(7-(8 - chloronaphthalen - 1 - yl)-2 - (((2R,7aS)-2 - fluorotetrahydro - 1H - pyrrolizin - 7a(5H)-yl)methoxy)-1,5 - naphthyridin - 4 - yl)-2-(cyanomethyl)piperazine - 1 - carboxylate (42; 388 mg, 0.58 mmol) in CH 2 Cl 2 (10 mL) to form a viscous solid, and the mixture was stirred at room temperature. After 2.5 h, the solid was sampled and determined to be the product by LC / MS. The DCM layer was decanted off, and then the viscous solid was dissolved in MeOH. The solution was diluted with saturated NaHCO 3 aqueous solution, extracted with DCM (3x), dried (MgSO 4 ), filtered, and concentrated in vacuo. The crude product was purified by silica gel column chromatography, eluting with a 0% to 10% gradient of MeOH in DCM containing 10% NH 4 OH (v / v) to give 198 mg (60%) of 2 - ((S)-4-(7-(8 - chloronaphthalen - 1 - yl)-2 - (((2R,7aS)-2 - fluorotetrahydro - 1H - pyrrolizin - 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; 1 H NMR (300 MHz, DMSO-d 6 ) δ 8.58 (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).

[0195] 2 - ((S)-1 - acryloyl - 4 - (7 - (8 - chloronaphthalen - 1 - yl)-2 - (((2R,7aS)-2 - fluorotetrahydro - 1H - pyrrolizin - 7a(5H)-yl)methoxy)-1,5 - naphthyridin - 4 - yl)piperazin - 2 - yl)acetonitrile (1ag). 2 - ((S)-4 - (7 - (8 - chloronaphthalen - 1 - yl)-2 - (((2R,7aS)-2 - fluorotetrahydro - 1H - pyrrolizin - 7a(5H)-yl)methoxy)-1,5 - naphthyridin - 4 - yl)piperazin - 2 - yl)acetonitrile (43; 90 mg, 0.16 mmol) in CH 2 Cl 2 (12 mL) was treated with Et 3 N (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 Et 3 N and 3.8 μL of acryloyl chloride were added, and the mixture was stirred in an ice bath. After 1 h, the mixture was diluted with CH 2 Cl 2 , washed with H 2 O (2X), dried (MgSO 4 ), filtered, and concentrated in vacuo. The crude product was purified by silica gel column chromatography (eluting with a 0% to 10% gradient of MeOH in DCM) to afford 73 mg (74%) of 2 - ((S)-1 - acryloyl - 4 - (7 - (8 - chloronaphthalen - 1 - yl)-2 - (((2R,7aS)-2 - fluorotetrahydro - 1H - pyrrolizin - 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; 1 H NMR (300 MHz, CDCl 3 ) δ 8.69 (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).

[0196] Example 8

[0197] 2 - ((S) - 4 - (7 - (8 - chloronaphthalen - 1 - yl) - 2 - (((2R,7aS) - 2 - fluorotetrahydro - 1H - pyrrolizin - 7a(5H) - yl)methoxy) - 1,5 - naphthyridin - 4 - yl) - 1 - (2 - fluoropropenoyl)piperazin - 2 - yl)acetonitrile (1ah).

[0198]

[0199] Example 8 (1ah) was prepared as shown in Scheme 9 below.

[0200] Scheme 9

[0201]

[0202] 2 - ((S) - 4 - (7 - (8 - chloronaphthalen - 1 - yl) - 2 - (((2R,7aS) - 2 - fluorotetrahydro - 1H - pyrrolizin - 7a(5H) - yl)methoxy) - 1,5 - naphthyridin - 4 - yl) - 1 - (2 - fluoropropenoyl)piperazin - 2 - yl)acetonitrile (1ah). In N 2 atmosphere, propylphosphonic anhydride solution (T 3P (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-pyrrolizin-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 Et 3 N (0.12 mL, 0.80 mmol) in EtOAc (10 mL), and the mixture was stirred at room temperature to form a pale pink reaction mixture. After 1 h, the pale pink turned golden yellow. The mixture was stirred at room temperature and after 3 h, the reaction mixture was light brown. The mixture was diluted with EtOAc and washed with 5% aqueous NaHCO 3 solution (3X), dried (MgSO 4 ), filtered, and concentrated in vacuo. 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-pyrrolizin-7a(5H)-yl)methoxy)-1,5-naphthyridin-4-yl)-1-(2-fluoroprop-2-enoyl)piperazin-2-yl)acetonitrile (1ah) as an off-white solid: HPLC-MS (ES + ) m / z [M + H + = 643, 645; 1 H NMR (300 MHz, CDCl 3 ) δ 8.69 (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).

[0203] Nucleotide Exchange Assay

[0204] The biological activity of the examples was determined in KRAS.

[0205] The G12D / SOS1 nucleotide exchange assay was performed by Reaction Biology Corporation (RBC), 1 Great Valley Parkway, Suite 2 Malvern, PA 19355, USA. This assay evaluated the SOS1-mediated exchange of Bodipy-GDP to GTP observed with KRAS G12C and KRAS G12D.

[0206] The compounds were tested in 3-fold serial dilutions starting at 10 concentrations, with an initial concentration of 5 μM for ARS1620 and 10 μM for Examples 1-4, MRTX849, and MRTX1133. The compounds were pre-incubated at room temperature for 30 minutes, and curve fitting was performed when the activity of the highest concentration of the compound was less than 65%. 50 Reaction buffer: 40 mM HEPES 7.4, 10 mM MgCl

[0207] 2 2 , 1 mM DTT 0.002% Triton X100, 0.5% DMSO.

[0208] Enzyme: SOS1 (RBC cat# MSC-11-502). Recombinant human SOS1 (Genbank accession # NM_033360.3; aa564-1049, expressed in E. coli, with a C-terminal StrepII).

[0209] KRAS G12C and KRAS G12D: Recombinant human KRAS G12C or KRAS G12D (aa2-169, expressed in E. coli, with an N-terminal TEV-cleavable his-tag) pre-loaded with a 5-fold excess of Bodipy TM -GDP, and the excess Bodipy- TM GDP was removed from the loaded protein using a spin desalting column.

[0210] Final concentrations: KRAS-Bodipy TM -GDP at 0.125 μM; SOS1 at 750 nM; and GTP at 25 μM.

[0211] Reaction steps:

[0212] 1. Transfer 10 μL of a 1.5-fold KRAS solution (in freshly prepared reaction buffer) to the reaction wells.

[0213] 2. Transfer the compound in 100% DMSO into the buffer using acoustic technology (Echo550; nanoliter scale).

[0214] 3. Incubate the compound with KRAS at room temperature for 30 minutes.

[0215] 4. Prepare a 3-fold (SOS1 + GTP) solution in the reaction buffer.

[0216] 5. Transfer 5 μL of the SOS1 + GTP solution into the reaction wells (transfer only GTP into the first column as a control without SOS1).

[0217] 6. Monitor the reaction progress for 30 minutes at room temperature by monitoring the decrease in fluorescence signal using a PHERAstar (BMG Labtech plate reader (Ex / Em = 485 / 520)).

[0218] Data analysis: Normalize the fluorescence data using the following equation and fit it to the "one-phase exponential decay" equation using GraphPad Prism software. The plateau is fixed at zero (for non-covalent inhibitors), and the rate × 1000 is used to calculate the IC 50 value.

[0219]

[0220] where Yraw is defined as the fluorescence at time t, Ao is the average initial fluorescence without SOS1, and M is the minimum fluorescence at the end of the reaction at maximum SOS1.

[0221] The signal minus the background (the wells without SOS1 protein are used as the background) is converted to % activity relative to the DMSO control. Analyze the data using the "Sigmoidal dose-response (variable slope)" in GraphPad Prism 4; 4 parameters with a Hill slope. The constraints are bottom (constant equal to 0) and top (must be less than 120).

[0222] Results:

[0223]

[0224]

[0225] * Calculate the IC 50 value using the dRFU analysis method for covalent inhibitors, with Bodipy-GDP / KRAS G12C as the substrate and 0.5% DMSO in the reaction. ARS-1620 and MRTX-849 are reference standards for KRAS G12C.

[0226]

[0227] *IC 50 The value was analyzed by the rate constant method for reversible inhibitors (plateau = 0), using Bodipy-GDP / KRAS G12D as the substrate, and the reaction contained 0.5% DMSO. MRTX1133 was the reference standard for KRAS G12D.

[0228] KRAS G12C cell assay

[0229] The cellular activity of KRAS G12C was determined by Reaction Biology Corporation (RBC), 1 Great Valley Parkway, Suite 2 Malvern, PA 19355, USA in transiently transfected HEK293 cells by a target-binding cell assay (NanoBRET TM ). Prior to the assay, HEK293 cells were cultured to 70 - 80% confluence, then trypsinized and harvested. BI 2852 was used as the KRAS G12C reference 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 by an Echo 550.

[0230] A 10 μg / mL DNA solution was prepared in serum-free Opti-MEM, which consisted of 1 μg -KRAS(G12C)-NanoLuc fusion vector, 1 μg -KRAS(G12C)-NanoLuc fusion vector and 8 μg transfection vector DNA. Subsequently, 30 μL of FuGENE HD transfection reagent was added to each milliliter of the DNA mixture to form a lipid:DNA complex. The resulting mixture was gently inverted and incubated for 20 minutes at ambient temperature to allow complex formation. One part of the lipid:DNA complex was added to a mixture of 20 parts of suspended HEK293 cells in a sterile conical tube and gently inverted. Then the cell + lipid:DNA complex mixture was added to a sterile tissue culture dish and incubated for 24 hours. The medium in the dish was removed by aspiration, and then the cells were trypsinized to detach them from the tissue culture dish. Subsequently, the trypsin was neutralized with serum-containing medium and centrifuged at 200×g for 5 minutes to pellet the cells in a conical tube. The cell density was adjusted to 2×105 cells / mL in phenol red-free Opti-MEM. One part of the complete 20-fold concentrated NanoBRET TMThe RAS tracer reagent was dispensed into 20 aliquots of cells in conical tubes and gently inverted to mix. The resulting cell suspension was dispensed into a white 384-well NBS plate containing the test compound (starting at 10 μM, 10 doses, 3-fold dilutions) and incubated for 2 hours at 37 °C and 5% CO 2 2. The final concentration of the RAS tracer K2 was 1 μM. The NBS plate was removed from the incubator and equilibrated to room temperature for 15 minutes.

[0231] Freshly prepared substrate solution (3X) was added to each well of the 384-well NBS plate and incubated for 3 minutes at room temperature. The donor emission wavelength (460 nm) and acceptor emission wavelength (600 nm) were measured using an Envision 2104 plate reader. The raw BRET ratio was generated by dividing the acceptor emission value (600 nm) of each sample by the donor emission value (460 nm). To correct for background, the BRET ratio of the no-tracer (average of no-tracer control samples) was subtracted from the BRET ratio of each sample. The BRET ratio was calculated using the following equation: BRET ratio = [(acceptor sample emission value ÷ donor sample emission value) - (no-tracer control acceptor emission value ÷ no-tracer control donor emission value)]. The normalized BRET response (%) was calculated using the following equation: (BRET ratio of test compound / BRET ratio of DMSO control) × 100%. The IC 50 curve was plotted and the IC 50 value was calculated using GraphPad Prism 4 software based on the dose-response curve equation.

[0232] Results:

[0233]

[0234] *NanoBRET TM Target binding cell assay (KRAS G12C). BI-2852 is the reference standard for KRAS G12C.

[0235] CellTiter-Glo cell viability assay protocol

[0236] Materials:

[0237] The reference compound staurosporine was purchased from Sigma-Aldrich (Saint Louis, MO). The 2.0 luminescent cell viability detection 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 (fetal bovine serum), 2.5% horse serum, 100 μg / ml penicillin and 100 μg / ml streptomycin. The cultures were maintained in a humidified environment of 37 °C, 5% CO 2 and 95% air.

[0238] Steps:

[0239] 1. Dilute the test compound, reference compound, and staurosporine in DMSO solution with 10-dose and 3-fold dilutions in a source plate starting at 10 mM.

[0240] 2. Use an Echo 550 to transfer 25 nL of the test compound or 25 nL of staurosporine from the source plate to each well of a 384-well cell culture plate.

[0241] 3. Add 25 μL of medium containing 2000 cells to each well of the cell culture plate (in duplicate).

[0242] 4. Incubate the cells with the compound at 37 °C, 5% CO 2 for 72 hours.

[0243] 5. Add 25 μL of CellTiter-Glo 2.0 reagent to each well.

[0244] 6. Mix the contents on an orbital shaker for 2 minutes and then incubate at room temperature for 15 minutes to stabilize the luminescence signal.

[0245] 7. Record the luminescence intensity using an Envision 2104 multimode plate reader (PerkinElmer, Santa Clara, CA). Determine the number of viable cells in the culture based on the quantification of ATP present in each culture well.

[0246] 8. Using GraphPad Prism 4 software, plot the IC 50 curve based on the sigmoidal dose-response equation and calculate the IC 50 value.

[0247] Results:

[0248]

[0249] PK characteristics of male CD1 mice:

[0250]

[0251]

Claims

1. A compound of formula I: or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein, A is selected from an optionally hydrogenated, halogenated, hydroxylated, -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 an aryl or heteroaryl substituted by one or more of a 5- or 6-membered heterocycle; Y and G may be the same or different and are selected from hydrogen, halogen, C 1-4 alkyl, C 1-4 perdeuterated alkyl, -(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 key, O, S or NR 4 ; m is 0 - 2; n is 0 - 2; Z is C(R 4 ) 2 or is selected from a cyclic compound of a C 3-7 cycloalkyl group, a saturated or partially unsaturated 4- to 7-membered nitrogen-containing ring, or a saturated or partially unsaturated 7- to 10-membered nitrogen-bridged bicyclic ring; R 1 selected from 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 ); R 2 selected from hydrogen, -C(O)CH=CH, -C(O)CF=CH or -C(O)CCl=CH, provided that when R 2 is hydrogen, then m is 1 or 2; R 3 Selected from hydrogen, halogen, hydroxyl, -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 ; R 4 selected from hydrogen, C 1-4 alkyl, aryl or heteroaryl; R 5 selected from hydrogen, hydroxyl, C 1-4 alkyl, aryl, heteroaryl, C 1-4 alkoxy, aryloxy or heteroaryloxy; R 6 selected from N(R 4 ) 2 or a 4- to 7-membered saturated or unsaturated heterocycle containing one or more heteroatoms selected from N, O, and S.

2. The compound according to claim 1, wherein, the compound of formula I is selected from compounds 1a - 1ah and 2a - 2aw, including their pharmaceutically acceptable salts, solvates or prodrugs:

3. A pharmaceutical composition comprising the compound according to any one of claims 1 to 2, or a salt, solvate or prodrug thereof, and a pharmaceutically acceptable carrier.

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

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

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

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 mutants G12C or G12D.

8. The method of treating a disease, disorder or medical condition according to claim 5, wherein, the cancer is selected from glioma (glioblastoma), acute myeloid leukemia, acute myelocytic leukemia, myelodysplastic / myeloproliferative neoplasms, sarcoma, chronic myelomonocytic leukemia, non - Hodgkin 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 according to any one of claims 4 to 8, further comprising administering to the patient in need thereof at least one additional therapeutic agent.

10. The method according to claim 9, wherein, the additional therapeutic agent is selected from doxorubicin, paclitaxel, docetaxel, cisplatin, camptothecin, temozolomide, bevacizumab, trastuzumab, cetuximab, EGFR inhibitors, osimertinib, razertinib, CDK4 / 6 inhibitors, abemaciclib, palbociclib, ribociclib, c - MET inhibitors, capmatinib, volitinib, ALK inhibitors, crizotinib, alectinib, ceritinib, brigatinib, entrectinib, lorlatinib, PD - 1 antagonists, PD - L1 antagonists, ipilimumab, pembrolizumab or nivolumab.

11. The compound according to claim 2, wherein, the compound of formula I is selected from compounds 1ae - 1ah, 1b, 1d, 1i, 1q, 1s, 1x, 2z, 2al, 2an, 2ap and 2aq or a salt, solvate or prodrug thereof, 12. The pharmaceutical composition according to claim 3, comprising compound 1ae - 1ah, 1b, 1q or 2z or a salt, solvate or prodrug thereof, and a pharmaceutically acceptable carrier.

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

14. The method according to claim 13, wherein, the disease includes various cancers.

15. The method according to claim 14, wherein, the disease, disorder or medical condition is mediated by KRAS.

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

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

Images