WO2021113595A1 - Phosphorus derivatives as kras inhibitors - Google Patents

Abstract

The present invention is directed to inhibitors of Kirsten Rat sarcoma virus (KRAS), and more particularly to compounds of Formula (I) as well as compositions comprising Formula (I) and methods of using the compound of Formula (I) for the treatment or prevention of a disease, disorder, or medical condition mediated through KRAS, especially the KRAS mutant G12C.

Description

PHOSPHORUS DERIVATIVES AS KRAS INHIBITORS

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Application Ser. No. 62/944,571 filed December 6, 2019, which is incorporated herein in its entirety.

BACKGROUND OF THE INVENTION Field of the Invention

[0002] The present invention is directed to inhibitors of Kirsten Rat sarcoma virus (KRAS), and more particularly to phosphorous compounds, compositions and methods for the treatment or prevention of a disease, disorder, or medical condition mediated through KRAS, especially the KRAS mutant G12C. The diseases include various cancers.

Brief Description of the Related Art

[0003] Ras is a superfamily of small guanosine triphosphate (GTP) binding proteins consisting of various isoforms. Ras genes can mutate to oncogenes that are associated with numerous cancers such as lung, pancreas, and colon. Ras is one of the most frequently mutated oncogenes. KRAS, (Kirsten Rat sarcoma virus) an isoform of Ras, is one of the most frequently mutated Ras genes, comprising approximately 86% of all mutations. KRAS functions as an on/off switch in cell signaling. KRAS is a proto-oncogene that operates between inactive (GDP-bound) and active (GTP-bound) states to control a variety of functions, including cell proliferation. However, KRAS mutation leads to uncontrolled cell proliferation and cancer. KRAS-4B is the major isoform in cancers of the colon (30-40%), lung (15-20%) and pancreas (90%). (Liu, P. 2019, Acta Pharmaceutica Sinica B). Consequently, inhibitors of KRAS-GTP binding represent potential therapeutic agents for the treatment of various cancers.

[0004] Past attempts to design KRAS inhibitors have been mostly unsuccessful, due in large part to the high affinity of KRAS for GTP. However, more recent approaches that target the KRAS G12C mutation have shown more promise. This mutation exists in roughly 50% of lung cancers and approximately 10-20% of all KRAS G12 mutations. The cysteine residue of the mutation is positioned within the active site such that the sulfhydryl functionality can form a covalent bond with a suitably functionalized bound ligand (Liu, P. 2019, Acta Pharmaceutica Sinica B). This approach has identified irreversible, covalent inhibitors of the KRAS G12C mutation that are undergoing clinical study. Given the prominent role that KRAS plays as a driver of many malignancies, a need for new KRAS inhibitors with improved selectivity, safety, and efficacy exists. SUMMARY OF THE INVENTION In one aspect, the present invention is directed to a compound of Formula I:

or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein A and B are each independently chosen at each occurrence from aryl, or heteroaryl optionally substituted with one or more of hydrogen, halogen, hydroxyl, cyano, C_1-6alkyl, C₂- C6alkenyl, C2-C6alkynyl, -(C0-C6alkyl)cycloalkyl, C1-C6haloalkyl, and C1-6alkoxy; X is hydrogen, halogen or trifluoromethyl; R₁ is hydrogen, C₁-C₆alkyl, C_1-6alkoxy, -(C₀-C₆alkyl)cycloalkyl, C₁-C₆haloalkyl, -(C₁- C6alkyl)CN, -(C1-C6alkyl)P(O)R2R3; and R2 and R3 are each independently chosen at each occurrence from hydrogen, C1-C6alkyl, C₁-C₆alkoxyl, -(C₀-C₆alkyl)phenyl, -(C₀-C₆alkyl)aryl, -(C₀-C₆alkyl)heteroarylhydroxyl; and n is either 0 or 1 with the proviso that when n is 0 then R1 is limited to -(C1-C6alkyl)P(O)R2R3 and -(C1-C6alkyl)CN. [0005] In another aspect, the present invention is directed to a pharmaceutical composition comprising a compound or salt of Formula I together with a pharmaceutically acceptable carrier.

[0006] In another aspect, the present invention is directed to a method of treating a disease, disorder, or medical condition in a patient, comprising the step of providing to a patient in need thereof a therapeutic agent, wherein the therapeutic agent comprises the compound of Formula I or salt thereof.

[0007] These and other aspects will become apparent upon reading the following detailed description of the invention.

DETAILED DESCRIPTION OF THE INVENTION TERMINOLOGY

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

[0009] The terms “a” and “an” do not denote a limitation of quantity, but rather denote the presence of at least one of the referenced items. The term “or” means “and/or”. The terms “comprising,” “having,” “including,” and “containing” are to be construed as open-ended terms (i.e., meaning “including, but not limited to”).

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

[0011] All methods described herein can be performed in a suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”), is intended merely to better illustrate the invention and does not pose a limitation on the scope of the invention unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention as used herein. Unless defined otherwise, technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art of this disclosure. [0012] Furthermore, the disclosure encompasses all variations, combinations, and permutations in which one or more limitations, elements, clauses, and descriptive terms from one or more of the listed claims are introduced into another claim. For example, any claim that is dependent on another claim can be modified to include one or more limitations found in any other claim that is dependent on the same base claim. Where elements are presented as lists, e.g., in Markush group format, each subgroup of the elements is also disclosed, and any element(s) can be removed from the group. [0013] All compounds are understood to include all possible isotopes of atoms occurring in the compounds. Isotopes include those atoms having the same atomic number but different mass numbers and encompass heavy isotopes and radioactive isotopes. By way of general example, and without limitation, isotopes of hydrogen include tritium and deuterium, and isotopes of carbon include ¹¹C, ¹³C, and ¹⁴C. Accordingly, the compounds disclosed herein may include heavy or radioactive isotopes in the structure of the compounds or as substituents attached thereto. Examples of useful heavy or radioactive isotopes include ¹⁸F, ¹⁵N, ¹⁸O, ⁷⁶Br, ¹²⁵I and ¹³¹I. [0014] All Formulae disclosed herein include all pharmaceutically acceptable salts of such Formulae. [0015] The opened ended term “comprising” includes the intermediate and closed terms “consisting essentially of” and “consisting of.” [0016] The term “substituted” means that any one or more hydrogens on the designated atom or group is replaced with a selection from the indicated group, provided that the designated atom’s normal valence is not exceeded. Combinations of substituents and/or variables are permissible only if such combinations result in stable compounds or useful synthetic intermediates. A stable compound or stable structure is meant to imply a compound that is sufficiently robust to survive isolation from a reaction mixture, and subsequent formulation into an effective therapeutic agent. [0017] A dash (

) that is not between two letters or symbols is used to indicate a point of attachment for a substituent. [0018] “Alkyl” includes both branched and straight chain saturated aliphatic hydrocarbon groups, having the specified number of carbon atoms, generally from 1 to about 8 carbon atoms. The term C1-C6alkyl as used herein indicates an alkyl group having from 1, 2, 3, 4, 5, or 6 carbon atoms. Other embodiments include alkyl groups having from 1 to 8 carbon atoms, 1 to 4 carbon atoms or 1 or 2 carbon atoms, e.g. C1-C8alkyl, C1-C4alkyl, and C1-C2alkyl. When C0-Cn alkyl is used herein in conjunction with another group, for example, -C0-C2alkyl(phenyl), the indicated group, in this case phenyl, is either directly bound by a single covalent bond (C₀alkyl), or attached by an alkyl chain having the specified number of carbon atoms, in this case 1, 2, 3, or 4 carbon atoms. Alkyls can also be attached via other groups such as heteroatoms as in –O-C0- C₄alkyl(C₃-C₇cycloalkyl). Examples of alkyl include, but are not limited to, methyl, ethyl, n- propyl, isopropyl, n-butyl, 3-methylbutyl, t-butyl, n-pentyl, and sec-pentyl. [0019] “Alkoxy” is an alkyl group as defined above with the indicated number of carbon atoms covalently bound to the group it substitutes by an oxygen bridge (-O-). Examples of alkoxy include, but are not limited to, methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, 2- butoxy, t-butoxy, n-pentoxy, 2-pentoxy, 3- pentoxy, isopentoxy, neopentoxy, n-hexoxy, 2- hexoxy, 3-hexoxy, and 3- methylpentoxy. Similarly an “Alkylthio” or a “thioalkyl” group is an alkyl group as defined above with the indicated number of carbon atoms covalently bound to the group it substitutes by a sulfur bridge (-S-). Similarly, “alkenyloxy”, “alkynyloxy”, and “cycloalkyloxy” refer to alkenyl, alkynyl, and cycloalkyl groups, in each instance covalently bound to the group it substitutes by an oxygen bridge (-O-). [0020] “Halo” or “halogen” means fluoro, chloro, bromo, or iodo, and are defined herein to include all isotopes of same, including heavy isotopes and radioactive isotopes. Examples of useful halo isotopes include ¹⁸F, ⁷⁶Br, and ¹³¹I. Additional isotopes will be readily appreciated by one of skill in the art. [0021] “Haloalkyl” means both branched and straight-chain alkyl groups having the specified number of carbon atoms, substituted with 1 or more halogen atoms, generally up to the maximum allowable number of halogen atoms. Examples of haloalkyl include, but are not limited to, trifluoromethyl, difluoromethyl, 2-fluoroethyl, and penta-fluoroethyl. [0022] “Haloalkoxy” is a haloalkyl group as defined above attached through an oxygen bridge (oxygen of an alcohol radical). [0023] “Peptide” means a molecule which is a chain of amino acids linked together via amide bonds (also called peptide bonds).

[0024] “Pharmaceutical compositions” means compositions comprising at least one active agent, such as a compound or salt of Formula II, and at least one other substance, such as a carrier. Pharmaceutical compositions meet the U.S. FDA’s GMP (good manufacturing practice) standards for human or non-human drugs.

[0025] “Carrier” means a diluent, excipient, or vehicle with which an active compound is administered. A “pharmaceutically acceptable carrier” means a substance, e.g., excipient, diluent, or vehicle, that is useful in preparing a pharmaceutical composition that is generally safe, non-toxic and neither biologically nor otherwise undesirable, and includes a carrier that is acceptable for veterinary use as well as human pharmaceutical use. A “pharmaceutically acceptable carrier” includes both one and more than one such carrier.

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

[0027] “Providing” means giving, administering, selling, distributing, transferring (for profit or not), manufacturing, compounding, or dispensing.

[0028] “Treatment” or “treating” means providing an active compound to a patient in an amount sufficient to measurably reduce any disease symptom, slow disease progression or cause disease regression. In certain embodiments treatment of the disease may be commenced before the patient presents symptoms of the disease.

[0029] A “therapeutically effective amount” of a pharmaceutical composition means an amount effective, when administered to a patient, to provide a therapeutic benefit such as an amelioration of symptoms, decrease disease progression, or cause disease regression.

[0030] A “therapeutic compound” means a compound which can be used for diagnosis or treatment of a disease. The compounds can be small molecules, peptides, proteins, or other kinds of molecules.

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

[0032] Compounds of the Formulae disclosed herein may contain one or more asymmetric elements such as stereogenic centers, stereogenic axes and the like, e.g., asymmetric carbon atoms, so that the compounds can exist in different stereoisomeric forms. These compounds can be, for example, racemates or optically active forms including atropisomers. For compounds with two or more asymmetric elements, these compounds can additionally be mixtures of diastereomers. For compounds having asymmetric centers, all optical isomers in pure form and mixtures thereof are encompassed. In these situations, the single enantiomers, i.e., optically active forms can be obtained by asymmetric synthesis, synthesis from optically pure precursors, or by resolution of the racemates. Resolution of the racemates can also be accomplished, for example, by conventional methods such as crystallization in the presence of a resolving agent, or chromatography, using, for example a chiral HPLC column. All forms are contemplated herein regardless of the methods used to obtain them.

[0033] All forms (for example solvates, optical isomers, enantiomeric forms, polymorphs, free compound and salts) of an active agent may be employed either alone or in combination.

[0034] The term “chiral” refers to molecules, which have the property of non- superimposability of the mirror image partner.

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

[0036] A “diastereomer” is a stereoisomer with two or more centers of chirality and whose molecules are not mirror images of one another. Diastereomers have different physical properties, e.g., melting points, boiling points, spectral properties, and reactivities. Mixtures of diastereomers may separate 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.

[0037] “Enantiomers” refer to two stereoisomers of a compound, which are non- superimposable mirror images of one another. A 50:50 mixture of enantiomers is referred to as a racemic mixture or a racemate, which may occur where there has been no stereoselection or stereo specificity in a chemical reaction or process. [0038] 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. In describing an optically active compound, the prefixes D and L or R and S are used to denote the absolute configuration of the molecule about its chiral center(s). The prefixes d and l or (+) and (-) are employed to designate the sign of rotation of plane-polarized light by the compound, with (-) or 1 meaning that the compound is levorotatory. A compound prefixed with (+) or d is dextrorotatory. [0039] A “racemic mixture” or “racemate” is an equimolar (or 50:50) mixture of two enantiomeric species, devoid of optical activity. A racemic mixture may occur where there has been no stereoselection or stereospecificity in a chemical reaction or process. [0040] A “chelating group” or “chelator” is a ligand group which can form two or more separate coordinate bonds to a single central atom, which is usually a metal ion. Chelating groups as disclosed herein are organic groups which possess multiple N, O, or S heteroatoms, and have a structure which allows two or more of the heteroatoms to form bonds to the same metal ion. [0041] “Pharmaceutically acceptable salts” include derivatives of the disclosed compounds in which the parent compound is modified by making inorganic and organic, non- toxic, acid or base addition salts thereof. The salts of the present compounds can be synthesized from a parent compound that contains a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting free acid forms of these compounds with a stoichiometric amount of the appropriate base (such as Na, Ca, Mg, or K hydroxide, carbonate, bicarbonate, or the like), or by reacting free base forms of these compounds with a stoichiometric amount of the appropriate acid. Such reactions are typically carried out in water or in an organic solvent, or in a mixture of the two. Generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are used, where practicable. Salts of the present compounds further include solvates of the compounds and of the compound salts. [0042] Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like. The pharmaceutically acceptable salts include the conventional non-toxic salts and the quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. For example, conventional non- toxic acid salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like; and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, mesylic, esylic, besylic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, HOOC-(CH2)n-COOH where n is 0-4, and the like. Lists of additional suitable salts may be found, e.g., 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. [0043] The compounds of the present invention relate to phosphorus-containing 4-(4- acryloylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one derivatives, or a pharmaceutically acceptable salt, solvate, or prodrug thereof. The compound of Formula I is capable of forming an irreversible covalent bond with a cysteine residue within the active site of the KRAS mutant G12C, and inactivating it. One aspect of the present invention is directed to a phosphorus-containing-derivative of Formula I:

or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein A and B are each independently chosen at each occurrence from aryl, or heteroaryl optionally substituted with one or more of hydrogen, halogen, hydroxyl, cyano, C_1-6alkyl, C₂- C6alkenyl, C2-C6alkynyl, -(C0-C6alkyl)cycloalkyl, C1-C6haloalkyl, and C1-6alkoxy; X is hydrogen, halogen or trifluoromethyl; R₁ is hydrogen, C₁-C₆alkyl, C_1-6alkoxy, -(C₀-C₆alkyl)cycloalkyl, C₁-C₆haloalkyl, -(C₁- C6alkyl)CN, -(C1-C6alkyl)P(O)R2R3; and R2 and R3 are each independently chosen at each occurrence from hydrogen, C1-C6alkyl, C₁-C₆alkoxyl, -(C₀-C₆alkyl)phenyl, -(C₀-C₆alkyl)aryl, -(C₀-C₆alkyl)heteroarylhydroxyl; and n is either 0 or 1 with the proviso that when n is 0 then R1 is limited to -(C1-C6alkyl)P(O)R2R3. [0044] In another preferred embodiment of the compound of Formula I, A is heteroaryl optionally substituted with one or more of hydrogen, C1-6alkyl, and -(C0-C6alkyl)cycloalkyl; B is aryl optionally substituted with one or more of hydrogen, halogen, and hydroxyl; X is hydrogen, halogen or trifluoromethyl; R₁ is C₁-C₆alkyl, -(C₁-C₆alkyl)CN or -(C₁-C₆alkyl)P(O)R₂R₃; R₂ and R₃ are C₁-C₆alkyl or C₁-C₆alkoxyl and n is 1. [0045] In another preferred embodiment of the compound of Formula I, A is pyridyl, pyrimidinyl, pyrazolyl, or thiazolyl optionally substituted with one or more of hydrogen, C_1-6alkyl, C₂-C₆alkenyl, C₂-C₆alkynyl, -(C₀-C₆alkyl)cycloalkyl, C₁-C₆haloalkyl, and C_1-6alkoxy; B is aryl optionally substituted with one or more of hydrogen, halogen, hydroxyl, and cyano; X is hydrogen, halogen or trifluoromethyl; R1 is C1-C6alkyl, -(C1-C6alkyl)CN or -(C₁-C₆alkyl)P(O)R₂R₃; R₂ and R₃ are C₁-C₆alkyl or C₁-C₆alkoxyl; and n is 1. [0046] In another preferred embodiment of the compound of Formula I, A is pyridyl, pyrimidinyl, pyrazolyl, or thiazolyl optionally substituted with one or more of hydrogen, C_1-6alkyl, C₂-C₆alkenyl, C₂-C₆alkynyl, -(C₀-C₆alkyl)cycloalkyl, C₁-C₆haloalkyl, and C_1-6alkoxy; B is aryl optionally substituted with one or more of hydrogen, halogen, hydroxyl, and cyano; X is hydrogen, halogen or trifluoromethyl; R1 is -(C1-C6alkyl)P(O)R2R3R2 and R3 are C1-C6alkyl or C1-C6alkoxyl; and n is 0. [0047] In one particularly preferred embodiment of Formula I, ring A is phenyl, pyridyl, pyrimidinyl, pyrazolyl, or thiazolyl optionally substituted with one or more of hydrogen, C1-C4 alkyl, C1-C4 alkoxyl; ring B is a phenyl ring optionally substituted with one or more of hydrogen, halogen, hydroxyl, or cyano; X is hydrogen, halogen or trifluoromethyl; R¹ is hydrogen, C₁-C₆ alkyl, -(C1-C6alkyl)CN or -(C1-C6alkyl)P(O)R2R3; R2 and R3 are the same or different and are selected from hydrogen, C1-C6 alkyl, C1-C6 alkoxyl, or hydroxyl; and n is 1. Examples of particularly preferred embodiments of Formula I include the following, or pharmaceutically acceptable salts, solvates, or prodrugs thereof:^





[0048] Compounds disclosed herein can be administered as the neat chemical, but are preferably administered as a pharmaceutical composition. Accordingly, the invention encompasses pharmaceutical compositions comprising a compound or pharmaceutically acceptable salt of a compound, such as a compound of Formula I, together with at least one pharmaceutically acceptable carrier. The pharmaceutical composition may contain a compound or salt of Formula I as the only active agent, but is preferably contains at least one additional active agent. In certain embodiments the pharmaceutical composition is in a dosage form that 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 a 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 molar ratio of a compound, such as a compound of Formula I, and an additional active agent. For example the pharmaceutical composition may contain a molar ratio of about 0.5:1, about 1:1, about 2:1, about 3:1 or from about 1.5:1 to about 4:1 of an additional active agent to a compound of Formula I. [0049] Compounds disclosed herein may be administered orally, topically, parenterally, by inhalation or spray, sublingually, transdermally, via buccal administration, rectally, as an ophthalmic solution, or by other means, in dosage unit formulations containing conventional pharmaceutically acceptable carriers. The pharmaceutical composition may be formulated as any pharmaceutically useful form, e.g., as an aerosol, a cream, a gel, a pill, a capsule, a tablet, a syrup, a transdermal patch, or an ophthalmic solution. Some dosage forms, such as tablets and capsules, are subdivided into suitably sized unit doses containing appropriate quantities of the active components, e.g., an effective amount to achieve the desired purpose.

[0050] Carriers include excipients and diluents and must be of sufficiently high purity and sufficiently low toxicity to render them suitable for administration to the patient being treated. The carrier can be inert or it can possess pharmaceutical benefits of its own. The amount of carrier employed in conjunction with the compound is sufficient to provide a practical quantity of material for administration per unit dose of the compound.

[0051] Classes of carriers include, but are not limited to binders, buffering agents, coloring agents, diluents, disintegrants, emulsifiers, flavorants, glidants, lubricants, preservatives, stabilizers, surfactants, tableting agents, and wetting agents. Some carriers may be listed in more than one class, for example vegetable oil may be used as a lubricant in some formulations and a diluent in others. Exemplary pharmaceutically acceptable carriers include sugars, starches, celluloses, powdered tragacanth, malt, gelatin, talc, and vegetable oils. Optional active agents may be included in a pharmaceutical composition, which do not substantially interfere with the activity of the compound of the present invention.

[0052] The pharmaceutical compositions / combinations can be formulated for oral administration. These compositions contain between 0.1 and 99 weight % (wt.%) of a compound of Formula III and usually at least about 5 wt.% of a 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.

TREATMENT METHODS

[0053] The compounds of Formula I, as well as pharmaceutical compositions comprising the compounds, are useful for diagnosis or treatment of a disease, disorder, or medical condition mediated through KRAS, especially the KRAS mutant G12C, and including various cancers, such as glioma (glioblastoma), acute myelogenous leukemia, acute myeloid leukemia, myelodysplastic/myeloproliferative neoplasms, sarcoma, chronic myelomonocytic leukemia, non-Hodgkin lymphoma, astrocytoma, melanoma, non- small cell lung cancer, cholangiocarcinomas, chondrosarcoma, colon cancer or pancreatic cancer. Additional cancers that can be treated or diagnosed using the compounds of the invention include the following:

Cardiac: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma and teratoma; Lung: bronchogenic carcinoma (squamous cell, undifferentiated small cell, undifferentiated large cell, adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma, chondromatous hamartoma, mesothelioma; Gastrointestinal: esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumors, vipoma), small bowel (adenocarcinoma, lymphoma, carcinoid tumors, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large bowel (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma); Genitourinary tract: kidney (adenocarcinoma, Wilm's tumor (nephroblastoma), lymphoma, leukemia), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, interstitial cell carcinoma, fibroma, fibroadenoma, adenomatoid tumors, lipoma); Liver: hepatoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma; Biliary tract: gall bladder carcinoma, ampullary carcinoma, cholangiocarcinoma; Bone: osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulum cell sarcoma), multiple myeloma, malignant giant cell tumor chordoma, osteochronfroma (osteocartilaginous exostoses), benign chondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma and giant cell tumors; Nervous system: skull (osteoma, hemangioma, granuloma, xanthoma, osteitis deformans), meninges (meningioma, meningiosarcoma, gliomatosis), brain (astrocytoma, medulloblastoma, glioma, ependymoma, germinoma (pinealoma), glioblastoma multiform, oligodendroglioma, schwannoma, retinoblastoma, congenital tumors), spinal cord neurofibroma, meningioma, glioma, sarcoma); Gynecological: uterus (endometrial carcinoma (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma), granulosa-thecal cell tumors, Sertoli-Leydig cell tumors, dysgerminoma, malignant teratoma), vulva (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma (embryonal rhabdomyosarcoma), fallopian tubes (carcinoma); Hematologic: blood (myeloid leukemia (acute and chronic), acute lymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferative diseases, multiple myeloma, myelodysplastic syndrome), Hodgkin's disease, non-Hodgkin's lymphoma (malignant lymphoma); Skin: malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, moles dysplastic nevi, lipoma, angioma, dermatofibroma, keloids, psoriasis; and Adrenal glands: neuroblastoma.

[0054] According to the present invention, a method of KRAS-mediated diseases or conditions comprises providing to a patient in need of such treatment a therapeutically effective amount of a compound of Formula I. In one embodiment, the patient is a mammal, and more specifically a human. As will be understood by one skilled in the art, the invention also encompasses methods of treating non-human patients such as companion animals, e.g. cats, dogs, and livestock animals.

[0055] A therapeutically effective amount of a pharmaceutical composition is preferably an amount sufficient to reduce or ameliorate the symptoms of a disease or condition. In the case of KRAS-mediated diseases for example, a therapeutically effective amount may be an amount sufficient to reduce or ameliorate cancer. A therapeutically effective amount of a compound or pharmaceutical composition described herein will also provide a sufficient concentration of a compound of Formula I when administered to a patient. A sufficient concentration is preferably a concentration of the compound in the patient’s body necessary to prevent or combat the disorder. Such an amount may be ascertained experimentally, for example by assaying blood concentration of the compound, or theoretically, by calculating bioavailability.

[0056] According to the invention, the methods of treatment disclosed herein include providing certain dosage amounts of a compound of Formula I to a patient. Dosage levels of each compound of from about 0.1 mg to about 140 mg per kilogram of body weight per day are useful in the treatment of the above-indicated conditions (about 0.5 mg to about 7 g per patient per day). The amount of compound that may be combined with the carrier materials to produce a single dosage form will vary depending upon the patient treated and the particular mode of administration. Dosage unit forms will generally contain between from about 1 mg to about 500 mg of each active compound. In certain embodiments 25 mg to 500 mg, or 25 mg to 200 mg of a compound of Formula I are provided daily to a patient. Frequency of dosage may also vary depending on the compound used and the particular disease treated. However, for treatment of most KRAS-mediated diseases and disorders, a dosage regimen of 4 times daily or less can be used and in certain embodiments a dosage regimen of 1 or 2 times daily is used.

[0057] It will be understood, however, that the specific dose level for any particular patient will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, route of administration, and rate of excretion, drug combination and the severity of the particular disease undergoing therapy.

[0058] A compound of Formula I may be administered singularly (i.e., sole therapeutic agent of a regime) to treat or prevent KRAS-mediated diseases and conditions such as various canders, or may be administered in combination with another active agent. One or more compounds of Formula I may be administered in coordination with a regime of one or more other active agents such as anticancer cytotoxic agents. In an embodiment, a method of treating or diagnosing KRAS-mediated cancer in a mammal includes administering to said mammal a therapeutically effective amount of a compound of Formula I, optionally in combination with one or more additional active ingredients.

[0059] As will be appreciated by one skilled in the art, the methods of treatment provided herein are also useful for treatment of mammals other than humans, including for veterinary applications such as to treat horses and livestock, e.g. cattle, sheep, cows, goats, swine and the like, and pets (companion animals) such as dogs and cats.

[0060] For diagnostic or research applications, a wide variety of mammals will be suitable subjects including rodents (e.g. mice, rats, hamsters), rabbits, primates, and swine such as inbred pigs and the like. Additionally, for in vitro applications, such as in vitro diagnostic and research applications, body fluids (e.g. blood, plasma, serum, cellular interstitial fluid, saliva, feces, and urine) and cell and tissue samples of the above subjects will be suitable for use.

[0061] In one embodiment, the invention provides a method of treating a disease, disorder, or medical condition mediated through KRAS, especially the KRAS mutant G12C, including various cancers, in a patient identified as in need of such treatment, the method comprising providing to the patient an effective amount of a compound of Formula I. The compounds of Formula I provided herein may be administered alone, or in combination with one or more other active agents. [0062] In another embodiment, the method of treating or diagnosing KRAS-mediated diseases or conditions may additionally comprise administering the compound of Formula I in combination with one or more additional compounds, wherein at least one of the additional compounds is an active agent, to a patient in need of such treatment. The one or more additional compounds may include additional therapeutic compounds, including anticancer therapeutic compounds such as doxorubicin, paclitaxel, docetaxel, cisplatin, camptothecin, temozolomide, avastin, Herceptin, Erbitux, and the like. EXAMPLES Chemical Synthesis The compounds of the formula 1 described herein, and/or the pharmaceutically acceptable salts thereof, can be synthesized from commercially available starting materials by methods well known to those skilled in the art of synthetic organic chemistry. The following general synthetic Scheme 1 illustrates representative methods to prepare most of the example compounds. The listed starting materials, reactions, reagents, solvents, temperatures, catalysts and ligands are not limited to what is depicted for purely illustrative purposes. Certain abbreviations and acronyms well known to those trained in the art that may be used in Scheme 1 and in the Examples are listed below for clarity. Scheme 1 begins with the reaction of either an aromatic amine or a heteroaromatic amine A1 with bromine to afford the corresponding bromide B1. The reaction of commercially available 2,6-dichloro-5-fluoronicotinamide (C1) with oxalyl chloride generates an acyl isocyanate D1 that is not isolated, which after treatment with intermediate B1 furnishes the corresponding N-acylurea E1. Treatment of E1 with a strong base such as KHMDS results in cyclization to the pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione F1. Reaction of F1 with phosphorus oxychloride provides the chloro compound G1, which is subsequently reacted with^ N-Boc-piperazine H1 to yield intermediate I1. The Suzuki-Miyaura cross-coupling reaction of I1 with boron derivative such as potassium trifluoroborate salt J1 results in the coupled product K1. Palladium-mediated coupling of K1 with phosphine oxide L1 followed by deprotection with trifluoracetic acid yields the P-arylated amine derivative M1. Reaction of M1 with acryloyl chloride N1 provides a compound of the formula 1.

Scheme 1

Abbreviations and Acronyms The following abbreviations and acronyms may be used in this application: anhyd. = anhydrous; aq. = aqueous; B2pin2 = bis(pinacolato)diboron; Boc = tert-butoxycarbonyl; n-Bu₃P = tri-n-butylphosphine; CAS# = Chemical Abstracts Service Registry Number; Compd = compound; d = day(s); DCM = dichloromethane; DIEA = DIPEA = N,N-diisopropylethylamine; DMF = N,N-dimethylformamide; DMSO = dimethylsulfoxide; DMA = N,N-dimethylacetamide; dppf = 1,1'-bis(diphenylphosphino)ferrocene) EtOAc = ethyl acetate; Ex = Example; FCC = flash column chromatography using silica; h = hour(s); KHMDS = potassium bis(trimethylsilyl)amide [KN(SiMe₃)₂]; LDA = lithium diisopropylamide; LiHMDS = lithium bis(trimethylsilyl)amide [LiN(SiMe3)2]; MeOH = methanol; min = minutes; Pd2(dba)3 = tris(dibenzylideneacetone)dipalladium(0); Pd(dppf)Cl₂ = [1,1^-bis(diphenylphosphino)ferrocene]dichloropalladium(II); RT = room temperature; satd. = saturated solution; TEA = triethylamine; TFA = trifluoroacetic acid; THF = tetrahydrofuran; Xantphos = 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (CAS# 161265-03-8); EXAMPLES The following non-limiting Examples further illustrate certain aspects of the present invention. These compounds are prepared according to the general synthetic schemes described above. Example 1 4-((S)-4-Acryloyl-2-methylpiperazin-1-yl)-1-(4-(dimethylphosphoryl)-2-isopropyl-6- methylphenyl)-6-fluoro-7-(2-fluoro-6-hydroxyphenyl)pyrido[2,3-d]pyrimidin-2(1H)-one (1).

Example 1 was prepared as shown below in Scheme 2.

Scheme 2

N-((4-Bromo-2-isopropyl-6-methylphenyl)carbamoyl)-2,6-dichloro-5- fluoronicotinamide (1c). A solution of 2,6-dichloro-5-fluoronicotinamide (1a; CAS# 113237- 20-0; 1.56 g, 7.45 mmol) in THF (7.0 mL) was cooled to 5 °C while stirring under nitrogen. A solution of 2 M oxalyl chloride in DCM (2.03 mL, 4.07 mmol) was added dropwise to the reaction mixture over 10 min at 5 °C and then heated at 60 °C for 1 h. Subsequently, the reaction mixture was concentrated under a stream of nitrogen, diluted with THF (5.0 mL) and then cooled to 5 °C. A solution of 4-bromo-2-methyl-6-(1-methylethyl)benzenamine (1b; CAS# 773887-07-3; 1.70 g, 7.45 mmol) in THF (2.0 mL) was added dropwise over 5 min to reaction and stirred at 5 °C for 2 h. The crude reaction mixture was concentrated in vacuo and the residue was partitioned between 10% aq. citric acid (80 mL; w/v) and EtOAc (30 mL). The acidic aqueous layer was extracted again with EtOAc (2 x 30 mL) and the combined organic extracts were washed with brine (2 x 20 mL), dried (Na2SO4), filtered and concentrated in vacuo to give 3.26 g (95%) of N-((4-bromo-2-isopropyl-6-methylphenyl)carbamoyl)-2,6-dichloro-5- fluoronicotinamide (1c) as a tan foam: HPLC-MS (ES⁺) m/z MH⁺ = 464.07. 1-(4-Bromo-2-isopropyl-6-methylphenyl)-7-chloro-6-fluoropyrido[2,3-d]pyrimidine- 2,4(1H,3H)-dione (1d). A solution of N-((4-bromo-2-isopropyl-6-methylphenyl)carbamoyl)- 2,6-dichloro-5-fluoronicotinamide (1c; 3.20 g, 6.91 mmol) in anhyd. THF (12 mL) was cooled to 5 °C while stirring under nitrogen. A solution of KHMDS (1M) in THF (15.2 mL; 15.2 mmol) dropwise over 10 min at 5 °C and the resulting dark brown solution was stirred at RT for 2 h. The reaction was quenched by the addition of H₂O (10 mL) and concentrated in vacuo. The residue was partitioned between 10% aq. citric acid (80 mL) and EtOAc (30 mL). The acidic aqueous layer was extracted again with EtOAc (2 x 30 mL) and the combined organic extracts were washed with brine (2 x 30 mL), dried (Na₂SO₄), filtered and concentrated in vacuo. The crude product was purified by chromatography on silica gel eluting with gel eluting with a gradient of 15-50% EtOAc in hexane to afford 1.03 g (35%) of 1-(4-bromo-2-isopropyl-6- methylphenyl)-7-chloro-6-fluoropyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione (1d) as an off-white solid: HPLC-MS (ES-) m/z (M-1) = 424.04;

NMR (300 MHz, CDCl3): δ 8.66 (br s, 1H), 8.23 (d, J = 6.6 Hz, 1H), 7.43 (d, J = 2.1 Hz, 1H), 7.36 (d, J = 2.1 Hz, 1H), 2.53 (septet, J = 6.8 Hz, 1H), 2.03 (s, 3H), 1.17 (d, J = 6.8 Hz, 3H), 1.06 (d, J = 6.8 Hz, 3H). 1-(4-Bromo-2-isopropyl-6-methylphenyl)-4,7-dichloro-6-fluoropyrido[2,3- d]pyrimidin-2(1H)-one (1e). 1-(4-Bromo-2-isopropyl-6-methylphenyl)-7-chloro-6- fluoropyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione (1d) (703 mg, 1.65 mmol) and DIPEA (474 µL; 5.09 mmol) were combined at RT in anhyd. MeCN (16 mL). POCl₃ (254 µL; 2.73 mmol) was added the resulting yellow solution was heated at 80 °C for 2 h while stirring under nitrogen. The reaction mixture was cooled to RT and concentrated in vacuo and isolated under a nitrogen atmosphere. The resulting crude 1-(4-bromo-2-isopropyl-6-methylphenyl)-4,7-dichloro-6- fluoropyrido[2,3-d]pyrimidin-2(1H)-one (1e) was used directly in the next step: HPLC-MS (ES⁺) m/z MH⁺ = 443.83. tert-Butyl 4-(1-(4-bromo-2-isopropyl-6-methylphenyl)-7-chloro-6-fluoro-2-oxo-1,2- dihydropyrido[2,3-d]pyrimidin-4-yl)piperazine-1-carboxylate (1g). A solution of 1-(4- bromo-2-isopropyl-6-methylphenyl)-4,7-dichloro-6-fluoropyrido[2,3-d]pyrimidin-2(1H)-one (1e) (734 mg, 1.65 mmol) and DIPEA (1.44 mL, 8.24 mmol) in in anhyd. MeCN (16 mL) was cooled to 5 °C while stirring under nitrogen. tert-Butyl (S)-3-methylpiperazine-1-carboxylate (1f) (363 mg, 1.81 mmol) was added and the cooling bath was removed. After 1 h at RT the resulting amber solution was concentrated in vacuo and the residue was partitioned between 10% aq. citric acid (50 mL) and EtOAc (25 mL). The acidic aqueous layer was extracted again with EtOAc (2 x 25 mL) and the combined organic extracts were washed with brine (2 x 25 mL), dried (Na₂SO₄), filtered and concentrated in vacuo. The crude product was purified by chromatography on silica gel eluting with gel eluting with a gradient of 15-50% EtOAc in hexane to afford 673 mg (67%) of tert-butyl 4-(1-(4-bromo-2-isopropyl-6-methylphenyl)-7- chloro-6-fluoro-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)piperazine-1-carboxylate (1g) as a beige solid: HPLC-MS (ES⁺) m/z MH⁺ = 607.54;

NMR (300 MHz, CDCl₃): δ 7.7 (dd, J = 7.7, 0.8 Hz, 1H), 7.42-7.37 (m, 1H), 7.34-7.31 (m, 1H), 4.9-4.6 (m, 1H), 4.40-3.75 (ov m, 3H), 3.75-2.95 (ov m , 3H), 2.50-2.35 (m, 1H), 1.97 (1, 1.5H), 1.95 (s, 1.5H), 1.60-1.40 (ov, m, 12H), 1.16 (d, J = 6.8 Hz, 1.5 H), 1.15 (d, J = 6.8 Hz, 1.5 H), 1.04 (d, J = 6.8 Hz, 1.5 H), 1.02 (d, J = 6.8 Hz, 1.5 H). tert-Butyl (3S)-4-(1-(4-bromo-2-isopropyl-6-methylphenyl)-6-fluoro-7-(2-fluoro-6- hydroxyphenyl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1- carboxylate (1i). A slurry of 4-(1-(4-bromo-2-isopropyl-6-methylphenyl)-7-chloro-6-fluoro-2- oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)piperazine-1-carboxylate (1g) (200 mg, 0.328 mmol), (2-fluoro-6-hydroxyphenyl)boronic acid (1h) (62 mg, 0.393 mmol), Na2CO3 (104 mg, 0.984 mmol), 1,4-dioxane (4.24 mL) and water (0.75 mL) was degassed by sparging with nitrogen over 1 h. Tetrakis(triphenylphosphine)palladium(0) (38 mg, 0.033 mmol) was added and the reaction mixture was heated at 60 °C in a microwave reactor while stirring over 17 h. The resulting mixture was cooled to RT, concentrated in vacuo and partitioned between EtOAc (30 mL) and water (25 mL). The aqueous layer was extracted twice with EtOAc (2 x 20 mL) and the combined EtOAc extracts were dried, filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel eluting with a gradient of 30-70% EtOAc in hexane to furnish 96 mg (43%) of tert-butyl (3S)-4-(1-(4-bromo-2-isopropyl-6-methylphenyl)-6- fluoro-7-(2-fluoro-6-hydroxyphenyl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3- methylpiperazine-1-carboxylate (1i) as a yellow solid: HPLC-MS (ES⁺) m/z MH⁺ = 683.82; ¹H NMR (300 MHz, CDCl₃): δ 9.47 (s, 1H), 7.89 (dd, J = 9.6, 2.4 Hz, 1H), 7.47 (d, J = 1.8 Hz, 1H), 7.40 (d, J = 1.8 Hz, 1H),7.35-7.25 (m 1H), 6.75-6.50 (m, 2H), 5.10-4.60 (m, 1H), 4.60-3.00 (ov m, 6H), 2.58 (septet, J = 7.1 Hz, 1H), 2.00 (s, 1.5H), 1.97 (s, 1.5H), 1.65-1.40 (ov, m, 12H), 1.16 (d, J = 6.8 Hz, 1.5 H), 1.15 (d, J = 6.8 Hz, 1.5 H), 1.01 (d, J = 6.8 Hz, 1.5 H), 1.00 (d, J = 6.8 Hz, 1.5 H). 1-(4-(Dimethylphosphoryl)-2-isopropyl-6-methylphenyl)-6-fluoro-7-(2-fluoro-6- hydroxyphenyl)-4-((S)-2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (1j). Step 1. A slurry of tert-butyl (3S)-4-(1-(4-bromo-2-isopropyl-6-methylphenyl)-6-fluoro- 7-(2-fluoro-6-hydroxyphenyl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3- methylpiperazine-1-carboxylate (1i) (56 mg, 0.082 mmol), dimethylphosphine oxide (26 mg, 0.333 mmol) and triethylamine (68 µL, 0.488 mmol) in 1,4-dioxane (3.24 mL) was degassed by sparging with nitrogen over 1 h. Pd2(dba)3 (24 mg, 0.026 mmol) and Xantphos (15 mg, 0.026 mmol) were added and the reaction mixture was heated at 100 °C in a microwave reactor while stirring over 21 h. The resulting mixture was cooled to RT, concentrated in vacuo and partitioned between 10% aq. citric acid (15 mL; w/v) and EtOAc (10 mL). The acidic aqueous layer was extracted again with EtOAc (2 x 15 mL) and the combined organic extracts were washed with brine (2 x 10 mL), dried (Na₂SO₄), filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel eluting with a gradient of 0-10% MeOH in DCM to furnish 54 mg (71%) of tert-butyl (3S)-4-(1-(4-bromo-2-isopropyl-6-methylphenyl)-6-fluoro-7- (2-fluoro-6-hydroxyphenyl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine- 1-carboxylate: HPLC-MS (ES⁺) m/z MH⁺ = 682.11. Step 2. A solution of tert-butyl (3S)-4-(1-(4-bromo-2-isopropyl-6-methylphenyl)-6- fluoro-7-(2-fluoro-6-hydroxyphenyl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3- methylpiperazine-1-carboxylate (52 mg, 0.076 mmol) in DCM (2.0 mL) was treated with trifluoroacetic acid (400 µL) and stirred at RT under nitrogen for 3 h. The reaction mixture was concentrated in vacuo and partitioned between satd. aq. NaHCO3 (10 mL) and EtOAc (10 mL) . The aqueous layer was extracted again with EtOAc (2 x 15 mL) and the combined organic extracts were washed with brine (2 x 10 mL), dried (Na₂SO₄), filtered and concentrated in vacuo. The residue was purified by chromatography on silica gel eluting with a gradient of 5-15% MeOH in DCM to afford 31 mg (70%) of 1-(4-(dimethylphosphoryl)-2-isopropyl-6-methylphenyl)-6- fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-((S)-2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin- 2(1H)-one (1j): HPLC-MS (ES⁺) m/z MH⁺ = 582.05;

NMR (300 MHz, CDCl3): δ 9.40 (br s, 1H), 7.93 (dd, J = 9.7, 3.2 Hz, 1H), 7.79 (J = 12.3 Hz, 1H), 7.47 (d, J = 11.7 Hz, 1H), 7.35-7.20 7.40 (m, 1H), 6.75-6.55 (m, 2H), 5.10-4.65 (m, 1H), 4.50-4.20 (m, 1H), 3.80-3.55 (m, 1H), 3.30- 2.90 (ov m, 4H), 2.80-2.60 (m, 1H), 2.08 (s, 1.5H), 2.06 (s, 1.5H), 1.92 (br s, 1H), 1.80 (d, J = 13.0 Hz, 6H), 1.64 (d, J = 2.4 Hz, 1.5H), 1.59 (d, J = 2.4 Hz, 1.5H), 1.24 (d, J = 6.8 Hz, 1.5H), 1.23 (d, J = 6.8 Hz, 1.5 H), 1.05 (d, J = 6.8 Hz, 1.5 H), 1.03 (d, J = 6.8 Hz, 1.5 H). 2-(4-((S)-4-Acryloyl-2-methylpiperazin-1-yl)-1-(4-(dimethylphosphoryl)-2-isopropyl- 6-methylphenyl)-6-fluoro-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-7-yl)-3-fluorophenyl acrylate (1k). A solution of 1-(4-(dimethylphosphoryl)-2-isopropyl-6-methylphenyl)-6-fluoro- 7-(2-fluoro-6-hydroxyphenyl)-4-((S)-2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (1j) (31 mg, 0.053 mmol) and DIPEA (47 µL, 0.267 mmol) was dissolved in DCM (2.0 mL) and cooled to 5 °C while stirring under nitrogen. Acryloyl chloride (13 µL, 0.267 mmol) was added and the reaction mixture was stirred at 5 °C for 30 min and then quenched with 1 mL of water. The reaction mixture was diluted with DCM (10 mL) and extracted with satd. aq. NaHCO₃ (10 mL). The aqueous layer was extracted again with DCM (2 x 10 mL) and the combined organic extracts were washed with brine (2 x 10 mL), dried (Na₂SO₄), filtered and concentrated in vacuo of crude product that was a 7.3:1 mixture of 2-(4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-1-(4- (dimethylphosphoryl)-2-isopropyl-6-methylphenyl)-6-fluoro-2-oxo-1,2-dihydropyrido[2,3- d]pyrimidin-7-yl)-3-fluorophenyl acrylate (1k) (m/z MH⁺ = 690.02) and 1 (m/z MH⁺ = 636.09) as shown by HPLC-MS (ES⁺). This crude mixture was used without purification directly in the next step. 4-((S)-4-Acryloyl-2-methylpiperazin-1-yl)-1-(4-(dimethylphosphoryl)-2-isopropyl-6- methylphenyl)-6-fluoro-7-(2-fluoro-6-hydroxyphenyl)pyrido[2,3-d]pyrimidin-2(1H)-one (1). The crude 7.3:1 mixture of 1k (37 mg, 0.0533 mmol) and 1 from the previous step was dissolved in a mixture of 1,4-dioxane (1.60 mL) and water (0.40 mL) and then treated with 2M aq. LiOH (81 µL, 0.162 mmol) while stirring at RT under nitrogen. After 2 h, the pH was adjusted to pH 5 with 10% aq. citric acid and the reaction mixture was concentrated in vacuo. The residue was purified by chromatography on silica gel eluting with a gradient of 2-7% MeOH in DCM to afford 26 mg (77%) of 4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-1-(4-(dimethylphosphoryl)-2- isopropyl-6-methylphenyl)-6-fluoro-7-(2-fluoro-6-hydroxyphenyl)pyrido[2,3-d]pyrimidin- 2(1H)-one (1) as a light yellow powder: HPLC-MS (ES⁺) m/z MH⁺ = 636.09;

NMR (300 MHz, CDCl₃): δ 9.31 (s, 1H), 7.92 (dd, J = 9.5, 2.2 Hz, 1H), 7.79 (d, J = 12.3 Hz, 1H), 7.48 (d, J = 12.3 Hz, 1H), 7.35-7.25 (m, 1H), 6.75-6.40 (m, 3H), 6.42 (dd, J = 16.7, 1.8 Hz, 1H), 5.82 (dd, J = 10.4, 1.9 Hz, 1H), 5.3-3.4 (overlapping m, 6H), 3.40-2.90 (m, 1H), 2.80-2.50 (m, 1H), 2.15-2.0 (m, 3H), 1.80 (d, J = 13.1 Hz, 6H), 1.65-1.40 (m, 3H), 1.23 (d, J = 6.8 Hz, 1.5H), 1.22 (d, J = 6.8 Hz, 1.5H), 1.04 (d, J = 6.4 Hz, 1.5H), 1.03 (d, J = 6.4 Hz, 1.5H). Nucleotide Exchange Assay The biological activity of Example 1 was determined in a KRAS G12C/SOS1 Nucleotide Exchange Assay that was performed by Reaction Biology Corporation (RBC, 1 Great Valley Parkway, Suite 2 Malvern, PA 19355, USA). The assay evaluates the SOS1-mediated Bodipy- GDP to GTP exchange observed with KRAS G12C. The compounds were tested in 10 concentration IC50 mode with 3-fold serial dilution at a starting concentration of 10 ^M for Example 1 and 5 ^M for the reference standard (ARS-1620). The compound pre-incubation time was 30 min at RT and the curve fits were performed when the activities at the highest concentration of compounds were less than 65%. Reaction Buffer: 40 mM HEPES 7.4, 10 mM MgCl₂, 1 mM DTT 0.002% Triton X100, 0.1% DMSO (final). Enzyme: SOS1 (RBC cat# MSC-11-502). Recombinant human SOS1 (Genbank accession# NM_005633.3; aa 564-1049, expressed in E. Coli with C-terminal StrepII. MW=60.59 kDa). KRAS G12C: Recombinant human KRAS (Genbank accession# NM_033360.3; aa 2- 169, expressed in E. coli with N-terminal TEV cleavable his-tag. MW 21.4 kDa) KRAS is pre- loaded with Bodipy-GDP. Final concentrations: KRAS-bodipy-GDP was 0.125 µM, SOS1 was 70 nM and GTP was 25 µM. In addition, the final assay volume was 15 µL. Reaction Procedure: 1. Deliver 10 uL of 1.5x KRAS solution in freshly prepared reaction buffer to reaction wells. 2. Deliver compounds in 100% DMSO into buffer using acoustic technology (Echo550; nanoliter range). 3. Incubate compounds with KRAS for 30 minutes at room temperature. 4. Prepare 3x (SOS1 + GTP) solution in reaction buffer. 5. Deliver 5 µL of SOS1+GTP solution into reactions wells (deliver GTP only to column 1 for no SOS1 control). 6. Monitor reaction progress for 30 minutes at RT using a Clariostar plate reader (ex 483-14, ems 530-30). Data Analysis: The fluorescence data was normalized using the equation below and fitted to “one phase exponential decay” equation using GraphPad prism software. The plateau was fixed to zero (use for non-covalent inhibitors) and rate x1000 was used to calculate the IC50 values. Alternatively, the plateau was unconstrained (for covalent inhibitors) and the span value was used to calculate the IC₅₀ values.

where Yraw is defined as fluorescence at time t, Ao is the average initial fluorescence with no SOS1, and M is the minimum fluorescence at the end of the reaction at the maximum SOS1.

The background subtracted signals (no SOS1 protein wells were used as background) were converted to % activity relative to DMSO controls. Data was analyzed using GraphPad Prism 4 with “sigmoidal dose-response (variable slope)”; 4 parameters with Hill Slope. The constraints were bottom (constant equal to 0) and top (must be less than 1).

Results:

The present inventive concept has been described in terms of exemplary principles and embodiments, but those skilled in the art will recognize that variations may be made and equivalents substituted for what is described without departing from the scope and spirit of the disclosure as defined by the following claims.

Claims

CLAIMS WHAT IS CLAIMED IS: 1. A compound of Formula I:

or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein A and B are each independently chosen at each occurrence from aryl, or heteroaryl optionally substituted with one or more of hydrogen, halogen, hydroxyl, cyano, C_1-6alkyl, C₂- C₆alkenyl, C₂-C₆alkynyl, -(C₀-C₆alkyl)cycloalkyl, C₁-C₆haloalkyl, and C_1-6alkoxy; X is hydrogen, halogen or trifluoromethyl; R1 is hydrogen, C1-C6alkyl, C1-6alkoxy, -(C0-C6alkyl)cycloalkyl, C1-C6haloalkyl, -(C1- C₆alkyl)CN, -(C₁-C₆alkyl)P(O)R₂R₃; and R2 and R3 are each independently chosen at each occurrence from hydrogen, C1-C6alkyl, C1-C6alkoxyl, -(C0-C6alkyl)phenyl, -(C0-C6alkyl)aryl, -(C0-C6alkyl)heteroarylhydroxyl; and n is either 0 or 1 with the proviso that when n is 0 then R1 is limited to -(C1-C6alkyl)P(O)R2R3 and -(C1-C6alkyl)CN.

2. The compound or a pharmaceutically acceptable salt, solvate, or prodrug of claim 1 wherein A is aryl optionally substituted with one or more of hydrogen, C1-6alkyl, and -(C0- C₆alkyl)cycloalkyl; B is aryl optionally substituted with one or more of hydrogen, halogen, and hydroxyl; X is hydrogen, halogen or trifluoromethyl; R1 is C1-C6alkyl, -CH2CN or -CH2P(O)R2R3; R2 and R3 are C1-C6alkyl or C1-C6alkoxyl; and n is either 0 or 1 with the proviso that when n is 0 then R₁ is limited to -CH₂P(O)R₂R₃.

3. The compound or a pharmaceutically acceptable salt, solvate, or prodrug of claim 1 wherein A is heteroaryl optionally substituted with one or more of hydrogen, C1-6alkyl, and -(C0- C6alkyl)cycloalkyl; B is aryl optionally substituted with one or more of hydrogen, halogen, and hydroxyl; X is hydrogen, halogen or trifluoromethyl; R1 is C1-C6alkyl, -CH2CN; and -CH2P(O)R2R3; R2 and R3 are C1-C6alkyl or C1-C6alkoxyl; and n is either 0 or 1 with the proviso that when n is 0 then R₁ is limited to -CH₂P(O)R₂R₃.

4. The compound or a pharmaceutically acceptable salt, solvate, or prodrug of claim 3 wherein A is pyridyl, pyrimidinyl, pyrazolyl, or thiazolyl optionally substituted with one or more of hydrogen, C1-6alkyl, C2-C6alkenyl, C2-C6alkynyl, -(C0-C6alkyl)cycloalkyl, C1-C6haloalkyl, and C_1-6alkoxy; B is aryl optionally substituted with one or more of hydrogen, halogen, hydroxyl, and cyano; X1 is hydrogen or halogen; X2 is halogen or C1-6alkyl; R₁ is C₁-C₆alkyl, -CH₂CN, or -CH₂P(O)R₂R₃; and R₂ and R₃ are C₁-C₆alkyl or C₁-C₆alkoxyl; and n is either 0 or 1 with the proviso that when n is 0 then R1 is limited to -CH2P(O)R2R3.

5. The compound or a pharmaceutically acceptable salt, solvate, or prodrug of claim 1 wherein the compound of Formula I is:

^ ^ ^ ^







6. A pharmaceutical composition comprising a compound, salt, solvate, or prodrug of any one of claims 1 to 5, together with a pharmaceutically acceptable carrier.

7. A method of treating a disease, disorder, or medical condition in a patient, comprising the step of providing to a patient in need thereof a therapeutic agent, wherein the therapeutic agent is a compound, salt, solvate, or prodrug thereof of any one of claims 1 to 5.

8. The method of treating a disease, disorder, or medical condition in claim 7 in a patient, where diseases includes various cancers.

9. The method of treating a disease, disorder, or medical condition of claim 7 in a patient, where a disease, disorder, or medical condition is mediated through Kirsten Rat sarcoma virus (KRAS).

10. The method of treating a disease, disorder, or medical condition of claim 9 in a patient, where a disease, disorder, or medical condition is mediated through Kirsten Rat sarcoma virus (KRAS), especially the KRAS mutant G12C.

11. The method of claim 8, wherein the cancer is selected from glioma (glioblastoma), acute myelogenous leukemia, acute myeloid leukemia, myelodysplastic/myeloproliferative neoplasms, sarcoma, chronic myelomonocytic leukemia, non-Hodgkin lymphoma, astrocytoma, melanoma, non-small cell lung cancer, cholangiocarcinomas, chondrosarcoma, colon cancer or pancreatic cancer.

12. The method of claim 8, wherein the cancer is selected from Cardiac: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma and teratoma; Lung: bronchogenic carcinoma (squamous cell, undifferentiated small cell, undifferentiated large cell, adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma, chondromatous hamartoma, mesothelioma; Gastrointestinal: esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumors, vipoma), small bowel (adenocarcinoma, lymphoma, carcinoid tumors, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large bowel (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma); Genitourinary tract: kidney (adenocarcinoma, Wilm's tumor (nephroblastoma), lymphoma, leukemia), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, interstitial cell carcinoma, fibroma, fibroadenoma, adenomatoid tumors, lipoma); Liver: hepatoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma; Biliary tract: gall bladder carcinoma, ampullary carcinoma, cholangiocarcinoma; Bone: osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulum cell sarcoma), multiple myeloma, malignant giant cell tumor chordoma, osteochronfroma (osteocartilaginous exostoses), benign chondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma and giant cell tumors; Nervous system: skull (osteoma, hemangioma, granuloma, xanthoma, osteitis deformans), meninges (meningioma, meningiosarcoma, gliomatosis), brain (astrocytoma, medulloblastoma, glioma, ependymoma, germinoma (pinealoma), glioblastoma multiform, oligodendroglioma, schwannoma, retinoblastoma, congenital tumors), spinal cord neurofibroma, meningioma, glioma, sarcoma); Gynecological: uterus (endometrial carcinoma (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma), granulosa-thecal cell tumors, Sertoli-Leydig cell tumors, dysgerminoma, malignant teratoma), vulva (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma (embryonal rhabdomyosarcoma), fallopian tubes (carcinoma); Hematologic: blood (myeloid leukemia (acute and chronic), acute lymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferative diseases, multiple myeloma, myelodysplastic syndrome), Hodgkin's disease, non-Hodgkin's lymphoma (malignant lymphoma); Skin: malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, moles dysplastic nevi, lipoma, angioma, dermatofibroma, keloids, psoriasis; and Adrenal glands: neuroblastoma.

13. The method of any one of claims 7 to 11, further comprising administering to the patient in need thereof at least one additional therapeutic agent.