WO2025096608A1

WO2025096608A1 - Egfr covalent inhibition with macrocycles

Info

Publication number: WO2025096608A1
Application number: PCT/US2024/053656
Authority: WO
Inventors: Nathanael S. Gray; Zhengnian LI; Tinghu Zhang; Jianwei Che; Dimitrios GAZGALIS; Tyler BEYETT; Stephen Leycester GWALTNEY II
Original assignee: Dana Farber Cancer Institute Inc; Leland Stanford Junior University
Current assignee: Dana Farber Cancer Institute Inc; Leland Stanford Junior University
Priority date: 2023-10-31
Filing date: 2024-10-30
Publication date: 2025-05-08
Anticipated expiration: 2026-04-30

Abstract

The disclosure relates to macrocyclic compounds that act as inhibitors of epidermal growth factor receptor (EGFR); pharmaceutical compositions comprising the macrocyclic compounds; and methods of treating or preventing kinase-mediated disorders, including cancer and other proliferation diseases.

Description

EGFR Covalent Inhibition with Macrocycles

Cross-Reference to Related Applications

This application is claims priority to U.S. Provisional Application Serial No. 63/594,671 filed October 31 , 2023, which is hereby incorporated herein by reference in its entirety.

BACKGROUND

The epidermal growth factor receptor (EGFR, Erb-B1 ) belongs to a family of receptor tyrosine kinases that mediate the proliferation, differentiation, and survival of normal and malignant cells (Arteaga, C. L., J. Clin. Oncol. 19, 2001 , 32-40). Deregulation of EGFR has been implicated in many types of human cancer, with overexpression of the receptor present in at least 70% of human cancers (Seymour, L. K., Curr. Drug Targets 2, 2001 , 117-133), including non-small lung cell carcinomas, breast cancers, gliomas, squamous cell carcinomas of the head and neck, and prostate cancer (Raymond, E., et al., Drugs 60 (Suppl. 1 ), 2000, 15-23, discussion 41-2; Salomon, D. S., et al., Crit. Rev. Oncol. Hematol. 19, 1995, 183-232; Voldborg B. R., et al., Ann. Oncol. 8, 1997, 1197-1206). EGFR has therefore emerged as an attractive target for the design and development of diagnostic and therapeutic agents that can specifically bind and inhibit the receptor’s tyrosine kinase activity and signal transduction pathway in cancer cells. For example, the EGFR tyrosine kinase (EGFR-TK) reversible inhibitor TARCEVA® is approved by the FDA for treatment of NSCLC and advanced pancreatic cancer. Other anti-EGFR targeted molecules have also been approved, including Lapatinib and IRESSA®.

The epidermal growth factor receptor (EGFR) is one of the most investigated receptor protein tyrosine kinases and its link to non-small-cell lung cancer (NSCLC) is well established (A. Russo, T. et al., Oncotarget 2015, 6, 26814). However, over 75% of patients die five years after their NSCLC diagnosis. Tumors driven by activating mutations within the EGFR tyrosine kinase domain, e.g., point-mutation L858R or in-frame exon-19 deletions (ex19del) are initially sensitive to EGFR tyrosine kinase inhibitors (TKIs), such as gefitinib and erlotinib, (Paez, J. G., et al., Science (New York, N.Y.) 2004, 304, 1497; Lynch, T. J., et al., The New England Journal of Medicine 2004, 350, 2129), but these inhibitors are rendered resistant due to the acquisition of the secondary ‘gatekeeper’ T790M mutation (Pao, W., et al., PLoS Medicine 2005, 2, e73; Yu, H. A., et al., Clinical Cancer Research 2013, 19, 2240). Efforts to overcome first- generation TKI drug resistance resulted in the discovery and optimization of T790M-targeting irreversible inhibitors, which are rendered effective due to the ability to form covalent bonds with C797 (D. A. E. Cross, et al., Cancer Discovery 2014, 4, 1046; E. L. Kwak, et al., Proceedings of the National Academy of Sciences of the United States of America 2005, 702, 7665). Patients harboring T790M positive tumors respond well to treatment with AZD9291 , and more recently this drug has been shown to be a superior treatment as a front-line therapy in untreated EGFR mutant NSCLC patients (J.-C. Soria, et al., The New England Journal of Medicine 2018, 378, 113). However, despite these successes, patients can acquire resistance to AZD9291 through the acquisition of the C797S mutation that precludes the ability for the drug to form their essential covalent bonds (K. S. Thress, et al., Nature Medicine 2015, 27, 560).

Thus, there is a need for potent small molecule EGFR inhibitors with alternative mechanisms of action targeting mutant EGFR.

SUMMARY

In an aspect, provided herein is a compound of Formula (I):

or a pharmaceutically acceptable salt thereof, wherein the variables are defined herein.

In an embodiment, the compound of Formula (I) is a compound of Formula (IA):

In another embodiment, the compound of Formula (I) is a compound of Formula (IB):

In yet another embodiment, the compound of Formula (IA) is a compound of Formula (IIA-I):

In still another embodiment, the compound of Formula (IA) is a compound of Formula

(IA-II):

or a pharmaceutically acceptable salt thereof, wherein the variables are defined herein. In an embodiment, the compound of Formula (IA-II) is a compound of Formula (IA- lla):

In another embodiment, the compound of Formula (IA-II) is a compound of Formula (lA-llb):

In yet another embodiment, the compound of Formula (IA-II) is a compound of Formula (lA-llc):

In still another embodiment, the compound of Formula (lA-lla) is a compound of Formula (lA-llaa):

In an embodiment, the compound of Formula (lA-llb) is a compound of Formula (IA- llba):

In another embodiment, the compound of Formula (lA-llc) is a compound of Formula (lA-llca):

In yet another embodiment, the compound of Formula (lA-llc) is a compound of Formula (lA-llcb):

In another aspect, provided herein is a method of treating cancer or a proliferation disease, comprising administering to a subject in need thereof an effective amount of a compound disclosed herein or a pharmaceutical composition comprising a compound disclosed herein and a pharmaceutically acceptable carrier. In one embodiment, the cancer is lung cancer, breast cancer, glioma, squamous cell carcinoma, or prostate cancer. In another embodiment, the cancer is non-small cell lung cancer (NSCLC).

In yet another aspect, provided herein is a method of inhibiting the activity of EGFR, comprising administering to a subject in need thereof an effective amount of a compound of disclosed herein or a pharmaceutical composition comprising a compound disclosed herein and a pharmaceutically acceptable carrier. In an embodiment, the compound targets Cys775 on EGFR.

The disclosure also provides a kit comprising a compound capable of inhibiting EGFR activity selected from a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, and instructions for use in treating cancer. In one embodiment, the kit further comprises components for performing a test to determine whether a subject has an activating mutation in EGFR or a resistance mutation in EGFR.

DETAILED DESCRIPTION

Definitions

Listed below are definitions of various terms used to describe the compounds and compositions disclosed herein. These definitions apply to the terms as they are used throughout this specification and claims, unless otherwise limited in specific instances, either individually or as part of a larger group.

Unless defined otherwise, all technical and scientific terms used herein generally have the same meaning as commonly understood by one of ordinary skill in the art. Generally, the nomenclature used herein and the laboratory procedures in cell culture, molecular genetics, organic chemistry, and peptide chemistry are those well-known and commonly employed in the art.

As used herein, the articles “a” and “an” refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element. Furthermore, use of the term “including” as well as other forms, such as “include,” “includes,” and “included,” is not limiting.

As used herein, the term “about” will be understood by persons of ordinary skill in the art and will vary to some extent on the context in which it is used. As used herein when referring to a measurable value such as an amount, a temporal duration, and the like, the term “about” is meant to encompass variations of ±20% or ±10%, including ±5%, ±1%, and ±0.1% from the specified value, as such variations are appropriate to perform the disclosed methods.

The term “administration” or the like as used herein refers to the providing a therapeutic agent to a subject. Multiple techniques of administering a therapeutic agent exist in the art including, but not limited to, intravenous, oral, aerosol, parenteral, ophthalmic, pulmonary, and topical administration.

The term “treat,” “treated,” “treating,” or “treatment” includes the diminishment or alleviation of at least one symptom associated or caused by the state, disorder or disease being treated. In certain embodiments, the treatment comprises bringing into contact with wild-type or mutant EGFR an effective amount of a compound disclosed herein for conditions related to cancer.

As used herein, the term “prevent” or “prevention” means no disorder or disease development if none had occurred, or no further disorder or disease development if there had already been development of the disorder or disease. Also considered is the ability of one to prevent some or all of the symptoms associated with the disorder or disease.

As used herein, the term “patient,” “individual,” or “subject” refers to a human or a non-human mammal. Non-human mammals include, for example, livestock and pets, such as ovine, bovine, porcine, canine, feline and marine mammals. Preferably, the patient, subject, or individual is human.

As used herein, the terms “effective amount,” “pharmaceutically effective amount,” and “therapeutically effective amount” refer to a nontoxic but sufficient amount of an agent to provide the desired biological result. That result may be reduction or alleviation of the signs, symptoms, or causes of a disease, or any other desired alteration of a biological system. An appropriate therapeutic amount in any individual case may be determined by one of ordinary skill in the art using routine experimentation.

As used herein, the term “pharmaceutically acceptable” refers to a material, such as a carrier or diluent, which does not abrogate the biological activity or properties of the compound, and is relatively non-toxic, i.e., the material may be administered to an individual without causing undesirable biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained.

As used herein, the term “pharmaceutically acceptable salt” refers to derivatives of the disclosed compounds wherein the parent compound is modified by converting an existing acid or base moiety to its salt form. 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 of the present disclosure include the conventional nontoxic salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. The pharmaceutically acceptable salts of the present disclosure can be synthesized from the parent compound which contains a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two; generally, non-aqueous media like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. The phrase “pharmaceutically acceptable salt” is not limited to a mono, or 1 :1 , salt. For example, “pharmaceutically acceptable salt” also includes bis-salts, such as a bis-hydrochloride salt. Lists of suitable salts are found in Remington’s Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418 and Journal of Pharmaceutical Science, 66, 2 (1977), each of which is incorporated herein by reference in its entirety.

As used herein, the term “composition” or “pharmaceutical composition” refers to a mixture of at least one compound useful within the disclosure with a pharmaceutically acceptable carrier. The pharmaceutical composition facilitates administration of the compound to a patient or subject. Multiple techniques of administering a compound exist in the art including, but not limited to, intravenous, oral, aerosol, parenteral, ophthalmic, pulmonary, and topical administration.

The term “pharmaceutical combination” as used herein means a product that results from the mixing or combining of more than one active ingredient and includes both fixed and non-fixed combinations of the active ingredients. The term “fixed combination” means that the active ingredients, e.g., a compound of the disclosure and a co- agent, are both administered to a patient simultaneously in the form of a single entity or dosage. The term “non-fixed combination” means that the active ingredients, e.g. a compound of the disclosure and a co-agent, are both administered to a patient as separate entities either simultaneously, concurrently or sequentially with no specific time limits, wherein such administration provides therapeutically effective levels of the two compounds in the body of the patient. The latter also applies to cocktail therapy, e.g., the administration of three or more active ingredients.

As used herein, the term “pharmaceutically acceptable carrier” means a pharmaceutically acceptable material, composition or carrier, such as a liquid or solid filler, stabilizer, dispersing agent, suspending agent, diluent, excipient, thickening agent, solvent or encapsulating material, involved in carrying or transporting a compound useful within the disclosure within or to the patient such that it may perform its intended function. Typically, such constructs are carried or transported from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation, including the compound useful within the disclosure, and not injurious to the patient. Some examples of materials that may serve as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; surface active agents; alginic acid; pyrogen-free water; isotonic saline; Ringer’s solution; ethyl alcohol; phosphate buffer solutions; and other non-toxic compatible substances employed in pharmaceutical formulations.

As used herein, the term “EGFR” refers to epidermal growth factor receptor (alternately referred to as ErbB-1 or HER1) and may refer to the wild-type receptor or to a receptor containing one or more mutations.

As used herein, the term “HER” or Her” refers to members of the ErbB receptor tyrosine kinase family, including EGFR, ERBB2, HER3, and HER4.

As used herein, the term “allosteric site” refers to a site on EGFR other than the ATP binding site, such as that characterized in a crystal structure of EGFR. An “allosteric site” can be a site that is close to the ATP binding site, such as that characterized in a crystal structure of EGFR. For example, one allosteric site includes one or more of the following amino acid residues of epidermal growth factor receptor (EGFR): Lys745, Leu788, Ala743, Cys755, Leu777, Phe856, Asp855, Met766, Ile759, Glu762, and/or Ala763. As used herein, the term “agent that prevents EGFR dimer formation,” or iterations thereof, refers to an agent that prevents dimer formation in which the C-lobe of the “activator” subunit impinges on the N-lobe of the “receiver” subunit. Examples of agents that prevent EGFR dimer formation include, but are not limited to, cetuximab, trastuzumab, panitumumab, and Mig6.

As used herein, the term “alkyl,” by itself or as part of another substituent means, unless otherwise stated, a straight or branched chain hydrocarbon having the number of carbon atoms designated (i.e., C_1-6 alkyl means an alkyl having one to six carbon atoms) and includes straight and branched chains. Examples include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert butyl, pentyl, neopentyl, and hexyl. Other examples of C_1-6 alkyl include ethyl, methyl, isopropyl, isobutyl, n-pentyl, and n-hexyl.

As used herein, the term “haloalkyl” refers to an alkyl group, as defined above, substituted with one or more halo substituents, wherein alkyl and halo are as defined herein. Haloalkyl includes, by way of example, chloromethyl, trifluoromethyl, bromoethyl, chlorofluoroethyl, and the like.

As used herein, the term “alkoxy” refers to the group -O-alkyl, wherein alkyl is as defined herein. Alkoxy includes, by way of example, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, t-butoxy and the like.

The term “alkylcarbonyl” represents an alkyl group as defined above having the indicated number of carbon atoms bonded to a carbonyl group. Nonlimiting examples of C_1-6 alkylcarbonyl include hexanyl-C(O), tertbutyl-C(O), and isopropyl-C(O).

As used herein, the term “alkylcarbonyl,” means, unless otherwise stated, a straight or branched chain hydrocarbon having the number of carbon atoms designated (i.e., C_1-6 alkylcarbonyl means an alkyl having one to six carbon atoms, where one of the carbon atoms corresponds to a carbonyl, i.e., C=O) and includes straight and branched chains. In a nonlimiting example, C_1-6 alkylcarbonyl refers to Examples include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert butyl, pentyl, neopentyl, and hexyl. Other examples of C_1-6 alkyl include ethyl, methyl, isopropyl, isobutyl, n-pentyl, and n-hexyl.

As used herein, the term “alkenyl” refers to a monovalent group derived from a hydrocarbon moiety containing, in certain embodiments, from two to six, or two to eight carbon atoms having at least one carbon-carbon double bond. The alkenyl group may or may not be the point of attachment to another group. The term “alkenyl” includes, but is not limited to, ethenyl, 1 -propenyl, 1-butenyl, heptenyl, octenyl and the like.

As used herein, the term “halo” or “halogen” alone or as part of another substituent means, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom, preferably, fluorine, chlorine, or bromine, more preferably, fluorine or chlorine. As used herein, the term “cycloalkyl” means a non-aromatic carbocyclic system that is fully saturated having 1 , 2 or 3 rings wherein such rings may be fused. The term “fused” means that a second ring is present (i.e., attached or formed) by having two adjacent atoms in common (i.e., shared) with the first ring. Cycloalkyl also includes bicyclic structures that may be bridged or spirocyclic in nature with each individual ring within the bicycle varying from 3-8 atoms. In an embodiment, “cycloalkyl” is C3-C10 cycloalkyl. The term “cycloalkyl” includes, but is not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclo[3.1 ,0]hexyl, spiro[3.3]heptanyl, and bicyclo[1 .1.1 ]pentyl.

As used herein, the term “bicyclic ring” means a fused ring system comprising two rings, wherein the first ring is aryl or heteroaryl and the second ring is cycloalkyl or heterocycloalkyl. The term “bicyclic ring” includes, but is not limited to, indoline, isoindoline- 1 ,3-dione, isoindolin-1-one, and dihydro-indene. In an embodiment, the bicyclic ring is indoline.

As used herein, the term “heterocyclyl” or “heterocycloalkyl” means a non-aromatic carbocyclic system containing 1 , 2, 3 or 4 heteroatoms selected independently from N, O, and S and having 1 , 2 or 3 rings wherein such rings may be fused, wherein fused is defined above. In an embodiment, “heterocyclyl” or “heterocycloalkyl” is 3-10 membered heterocycloalkyl. Heterocyclyl also includes bicyclic structures that may be bridged or spirocyclic in nature with each individual ring within the bicycle varying from 3-8 atoms, and containing 0, 1 , or 2 N, O, or S atoms. The term “heterocyclyl” includes cyclic esters (i.e., lactones) and cyclic amides (i.e., lactams) and also specifically includes, but is not limited to, epoxidyl, oxetanyl, tetrahydro-furanyl, tetrahydropyranyl (i.e., oxanyl), pyranyl, dioxanyl, aziridinyl, azetidinyl, pyrrolidinyl, 2,5-dihydro-1 H-pyrrolyl, oxazolidinyl, thiazolidinyl, piperidinyl, morpholinyl, piperazinyl, thiomorpholinyl, 1 ,3-oxazinanyl, 1 ,3-thiazinanyl, 2- azabicyclo[2.1 ,1]hexanyl, 5-azabicyclo-[2.1 ,1]hexanyl, 6-azabicyclo[3.1.1] heptanyl, 2- azabicyclo[2.2.1]heptanyl, 3-azabicyclo[3.1 ,1]-heptanyl, 2-azabicyclo[3.1 ,1]heptanyl, 3- azabicyclo[3.1 .0]hexanyl, 2-azabicyclo[3.1 .0]hexanyl, 3-azabicyclo[3.2.1]octanyl, 8- azabicyclo[3.2.1]octanyl, 3-oxa-7-azabicyclo[3.3.1]nonanyl, 3-oxa-9- azabicyclo[3.3.1 ]nonanyl, 2-oxa-5-azabicyclo[2.2.1 ]heptanyl, 6-oxa-3-azabicyclo[3.1.1]- heptanyl, 2-azaspiro[3.3]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl, 2-oxaspiro[3.3]heptanyl, 2- oxaspiro[3.5]nonanyl, 3-oxaspiro[5.3]nonanyl, and 8-oxabicyclo[3.2.1]octanyl.

As used herein, the term “heterocycloalkenyl” refers to monounsaturated or polyunsaturated monocyclic carbocycles containing at least one heteroatom selected from oxygen, sulfur, and nitrogen and may comprise benzo-fused analogues thereof. The term “heterocycloalkenyl” specifically includes, but is not limited to dihydropyranyl, dihydrothiopyranyl, dihydrothiophenyl, and tetrahydropyridinyl. As used herein, the term “aromatic” refers to a carbocycle or heterocycle with one or more polyunsaturated rings and having aromatic character, i.e., having (4n + 2) delocalized IT (pi) electrons, where n is an integer.

As used herein, the term “aryl” means an aromatic carbocyclic system containing 1 , 2 or 3 rings, wherein such rings may be fused, wherein fused is defined above. If the rings are fused, one of the rings must be fully unsaturated and the fused ring(s) may be fully saturated, partially unsaturated or fully unsaturated. In an embodiment, “aryl” is C₆-C₁₀ aryl. The term “aryl” includes, but is not limited to, phenyl, naphthyl, indanyl, and 1 , 2,3,4- tetrahydronaphthalenyl. In some embodiments, aryl groups have 6 carbon atoms. In some embodiments, aryl groups have from six to ten carbon atoms. In some embodiments, aryl groups have from six to sixteen carbon atoms.

As used herein, the term “heteroaryl” means an aromatic carbocyclic system containing 1 , 2, 3, or 4 heteroatoms selected independently from N, O, and S and having 1 , 2, or 3 rings wherein such rings may be fused, wherein fused is defined above. In an embodiment, “heteroaryl” is 5-10 membered heteroaryl. The term “heteroaryl” includes, but is not limited to, furanyl, thienyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, thiadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, imidazo[1 ,2-a]pyridinyl, pyrazolo[1 ,5-a]pyridinyl, 5,6,7,8-tetrahydroisoquinolinyl, 5, 6,7,8- tetrahydroquinolinyl, 6,7-dihydro-5H-cyclopenta[b]pyridinyl, 6,7-dihydro-5H-cyclopenta- [c]pyridinyl, 1 ,4,5,6-tetrahydrocyclopenta[c]pyrazolyl, 2, 4,5,6- tetrahydrocyclopenta[c]pyrazolyl, 5,6-dihydro-4H-pyrrolo[1 ,2-b] pyrazolyl, 6,7-dihydro-5H- pyrrolo[1 ,2-b][ 1 ,2,4]triazolyl, 5,6,7,8-tetrahydro-[1 ,2,4]triazolo[1 ,5-a]pyridinyl, 4, 5,6,7- tetrahydropyrazolo[1 ,5-a]pyridinyl, 4,5,6,7-tetrahydro-1H-indazolyl and 4,5,6,7-tetrahydro- 2H-indazolyl.

It is to be understood that if an aryl, heteroaryl, cycloalkyl, bicyclic ring, or heterocyclyl moiety may be bonded or otherwise attached to a designated moiety through differing ring atoms (i.e., shown or described without denotation of a specific point of attachment), then all possible points are intended, whether through a carbon atom or, for example, a trivalent nitrogen atom. For example, the term “pyridinyl” means 2-, 3- or 4- pyridinyl, the term “thienyl” means 2- or 3-thienyl, and so forth.

As used herein, the term “substituted” means that an atom or group of atoms has replaced hydrogen as the substituent attached to another group.

As used herein, the term “optionally substituted” means that the referenced group may be substituted or unsubstituted. In one embodiment, the referenced group is optionally substituted with zero substituents, i.e., the referenced group is unsubstituted. In another embodiment, the referenced group is optionally substituted with one or more additional group(s) individually and independently selected from groups described herein. The term “cancer” includes, but is not limited to, the following cancers: breast, ovary, cervix, prostate, testis, genitourinary tract, esophagus, larynx, glioblastoma, neuroblastoma, stomach, skin, keratoacanthoma, lung, epidermoid carcinoma, large cell carcinoma, small cell carcinoma, lung adenocarcinoma, bone, colon, colorectal, adenoma, pancreas, adenocarcinoma, thyroid, follicular carcinoma, undifferentiated carcinoma, papillary carcinoma, seminoma, melanoma, sarcoma, bladder carcinoma, liver carcinoma and biliary passages, kidney carcinoma, myeloid disorders, lymphoid disorders, Hodgkin's, hairy cells, buccal cavity and pharynx (oral), lip, tongue, mouth, pharynx, small intestine, colon, rectum, large intestine, rectum, brain and central nervous system, chronic myeloid leukemia (CML), and leukemia. The term “cancer” includes, but is not limited to, the following cancers: myeloma, lymphoma, or a cancer selected from gastric, renal, head and neck, oropharangeal, non-small cell lung cancer (NSCLC), endometrial, hepatocarcinoma, nonHodgkin’s lymphoma, and pulmonary.

Additional cancers that the compounds described herein may be useful in preventing, treating and studying are, for example, colon carcinoma, familial adenomatous polyposis carcinoma and hereditary non-polyposis colorectal cancer, or melanoma. Further, cancers include, but are not limited to, labial carcinoma, larynx carcinoma, hypopharynx carcinoma, tongue carcinoma, salivary gland carcinoma, gastric carcinoma, adenocarcinoma, thyroid cancer (medullary and papillary thyroid carcinoma), renal carcinoma, kidney parenchyma carcinoma, cervix carcinoma, uterine corpus carcinoma, endometrium carcinoma, chorion carcinoma, testis carcinoma, urinary carcinoma, melanoma, brain tumors such as glioblastoma, astrocytoma, meningioma, medulloblastoma and peripheral neuroectodermal tumors, gall bladder carcinoma, bronchial carcinoma, multiple myeloma, basalioma, teratoma, retinoblastoma, choroidea melanoma, seminoma, rhabdomyosarcoma, craniopharyngeoma, osteosarcoma, chondrosarcoma, myosarcoma, liposarcoma, fibrosarcoma, Ewing sarcoma, and plasmocytoma.

Compounds

Provided herein are compounds that are inhibitors of epidermal growth factor receptor (EGFR) useful in the treatment of kinase-mediated disorders, including cancer and other proliferation diseases. In an embodiment, the compounds provided herein are mutant selective EGFR inhibitors.

In an aspect, provided herein is a compound of Formula (I):

or a pharmaceutically acceptable salt thereof, wherein:

Ring A and Ring B are each independently selected from the group consisting of absent, C_3-11 cycloalkyl, C_6-10 aryl, 3-11 membered heterocycloalkyl, 5-11 membered heteroaryl, and 6-11 membered bicyclic ring;

Ring C is absent or phenyl, provided that when C is absent, n is 0;

X¹ is N or CH;

X² is N, CH, or CR⁸;

Z is -(CR⁵R⁶)_s-, wherein 1-6 CR⁵R⁶ are optionally replaced with O, C(O), N(H), N(C_1-6 alkyl), N(C_1-6 haloalkyl), N(C_1-6 alkylcarbonyl), and wherein any two adjacent CR⁵R⁶ optionally combine to form -CR⁵=CR⁶- or -C≡C-;

R¹ is selected from the group consisting of halo, CN, OH, NH₂, N(H)(C_1-6 alkyl), N(C_1- ₆ alkyl)₂, C_1-6 alkyl, C_1-6 alkoxy, and C_1-6 haloalkyl;

R³ and R⁴ are each independently selected from the group consisting of halo, CN, OH, C_1-6 alkyl, C_1-6 alkoxy, C_1-6 haloalkyl, and C_1-6 alkylcarbonyl, wherein C_1-6 alkyl, C_1-6 alkoxy, C_1-6 haloalkyl, and C_1-6 alkylcarbonyl are optionally substituted with one or two R⁹;

R⁵ and R⁶ are each independently selected from the group consisting of H, halo, CN, OH, NH₂, N(H)(C_1-6 alkyl), N(C_1-6 alkyl)₂, C_1-6 alkyl, C_1-6 alkoxy, and C_1-6 haloalkyl;

R⁷, R⁸, and R⁹ are each independently selected from the group consisting of halo, CN, OH, NH₂, N(H)(C_1-6 alkyl), N(C_1-6 alkyl)₂, C_1-6 alkyl, C_1-6 alkoxy, and C_1-6 haloalkyl; each m, n, p, and r is independently 0, 1 , or 2; s is 1-15; is a single bond, a cis double bond, or a trans double bond; and

R² is represented by Formula (i-1):

wherein: L₃ is a bond, N(H), or N(C_1-6 alkyl); each R_E1, R_E2, and R_E3 is independently selected from the group consisting of H, halo, and C_1-6 alkyl; and

Y is O, S, or CH₂.

In embodiment, the compound of Formula (I) is a compound of Formula (IA):

or a pharmaceutically acceptable salt thereof, wherein:

X¹ is N or CH;

X² is N, CH, or CR⁸;

R⁷, R⁸, and R⁹ are each independently selected from the group consisting of halo, CN, OH, NH₂, N(H)(C_1-6 alkyl), N(C_1-6 alkyl)₂, C_1-6 alkyl, C_1-6 alkoxy, and C_1-6 haloalkyl; each m, n, p, and r is independently 0, 1 , or 2; s is 1-15; is a single bond, a cis double bond, or a trans double bond; and R² is represented by Formula (i-1):

Y is O, S, or CH₂.

or a pharmaceutically acceptable salt thereof, wherein:

X¹ is N or CH;

X² is N, CH, or CR⁸;

R⁵ and R⁶ are each independently selected from the group consisting of H, halo, CN, OH, NH₂, N(H)(C_1-6 alkyl), N(C_1-6 alkyl)₂, C_1-6 alkyl, C_1-6 alkoxy, and C_1-6 haloalkyl; R⁷, R⁸, and R⁹ are each independently selected from the group consisting of halo,

CN, OH, NH₂, N(H)(C_1-6 alkyl), N(C_1-6 alkyl)₂, C_1-6 alkyl, C_1-6 alkoxy, and C_1-6 haloalkyl; each m, p, and r is independently 0, 1 , or 2; s is 1-15; is a single bond, a cis double bond, or a trans double bond; and

R² is represented by Formula (i-1 ):

Y is O, S, or CH₂.

In yet another embodiment, the compound of Formula (IA) is a compound of Formula

or a pharmaceutically acceptable salt thereof, wherein:

Ring A and Ring B are each independently selected from the group consisting of absent, C_3-11 cycloalkyl, C_6-10 aryl, 3-1 1 membered heterocycloalkyl, 5-1 1 membered heteroaryl, and 6-1 1 membered bicyclic ring;

X¹ is N or CH;

R¹ is selected from the group consisting of halo, CN, OH, NH₂, N(H)(C_1-6 alkyl), N(C_1-

₆ alkyl)₂, C_1-6 alkyl, C_1-6 alkoxy, and C_1-6 haloalkyl; R³ and R⁴ are each independently selected from the group consisting of halo, CN, OH, C_1-6 alkyl, C_1-6 alkoxy, C_1-6 haloalkyl, and C_1-6 alkylcarbonyl, wherein C_1-6 alkyl, C_1-6 alkoxy, C_1-6 haloalkyl, and C_1-6 alkylcarbonyl are optionally substituted with one or two R⁹;

R⁹ is selected from the group consisting of halo, CN, OH, NH₂, N(H)(C_1-6 alkyl), N(C_1- ₆ alkyl)₂, C_1-6 alkyl, C_1-6 alkoxy, and C_1-6 haloalkyl; each n, p, and r is independently 0, 1 , or 2; s is 1-15; is a single bond, a cis double bond, or a trans double bond; and

R² is represented by Formula (i-1):

Y is O, S, or CH₂.

In another embodiment:

X¹ is N or CH;

X³ is N or CH, provided that when E is bonded to X³, then X³ is C; each X⁴ is independently N or CH; each X⁵ is independently NR¹¹ or CH₂, provided that when E is bonded to an X⁵, then said X⁵ is N or CH;

Z is -(CR⁵R⁶)_s-, wherein 1-6 CR⁵R⁶ are optionally replaced with O, C(O), N(H), N(C_1-6 alkyl), N(C_1-6 haloalkyl), N(C_1-6 alkylcarbonyl), and wherein any two adjacent CR⁵R⁶ optionally combine to form -CR⁵=CR⁶- or -C≡C-; each E is independently selected from the group consisting of a bond, CR⁵R⁶, O, C(O), and NR¹⁰;

R¹ is selected from the group consisting of halo, OH, C_1-6 alkyl, C_1-6 alkoxy, and C_1-6 haloalkyl; R³ and R⁴ are each independently selected from the group consisting of halo, CN, OH, C_1-6 alkyl, C_1-6 alkoxy, C_1-6 haloalkyl, and C_1-6 alkylcarbonyl, wherein C_1-6 alkyl, C_1-6 alkoxy, C_1-6 haloalkyl, and C_1-6 alkylcarbonyl are optionally substituted with one or two R⁹;

R⁵ and R⁶ are each independently selected from the group consisting of H, halo, CN, OH, C_1-6 alkyl, C_1-6 alkoxy, and C_1-6 haloalkyl; each R⁹ is independently selected from the group consisting of CN, NH₂, N(H)(C_1-6 alkyl), and N(C_1-6 alkyl)₂;

R¹⁰ is selected from the group consisting of H, C_1-6 alkyl, C_1-6 haloalkyl, and C_1-6 alkylcarbonyl;

R¹¹ is selected from the group consisting of C_1-6 alkyl, C_1-6 alkoxy, C_1-6 haloalkyl, and C_1-6 alkylcarbonyl; each n, p, and r is independently 0, 1 , or 2; s is 1-12; t and u are each independently 0, 1 , or 2; is a single bond, a cis double bond, or a trans double bond; and

R² is represented by Formula (i-1 ):

Y is O, S, or CH₂.

In yet another embodiment:

Ring A and Ring B are each independently selected from the group consisting of absent, C_6-10 cycloalkyl, C_6-10 aryl, 5-7 membered heterocycloalkyl, 5-7 membered heteroaryl, and 6-9 membered bicyclic ring;

X¹ is N or CH;

Z is -(CR⁵R⁶)_s-, wherein 1-4 CR⁵R⁶ are optionally replaced with O, C(O), N(H), N(C_1-6 alkyl), N(C_1-6 haloalkyl), N(C_1-6 alkylcarbonyl), and wherein any two adjacent CR⁵R⁶ optionally combine to form -CR⁵=CR⁶-;

R¹ is selected from the group consisting of C_1-6 alkyl, C_1-6 alkoxy, and C_1-6 haloalkyl; R³ and R⁴ are each independently selected from the group consisting of C_1-6 alkyl, C_1-

₆ alkoxy, C_1-6 haloalkyl, and C_1-6 alkylcarbonyl, wherein C_1-6 alkyl, C_1-6 alkoxy, C_1-6 haloalkyl, and C_1-6 alkylcarbonyl are optionally substituted with one R⁹;

R⁵ and R⁶ are each independently selected from the group consisting of H, halo, C_1-6 alkyl, C_1-6 alkoxy, and C_1-6 haloalkyl; each R⁹ is independently selected from the group consisting of NH₂, N(H)(C_1-6 alkyl), and N(C_1-6 alkyl)₂; each n, p, and r is independently 0, 1 , or 2; s is 1-10; is a single bond, a cis double bond, or a trans double bond; and

R² is represented by Formula (i-1 ):

Y is O, S, or CH₂.

In still another embodiment, the compound of Formula ( IA-I) is a compound of Formula (IA-II):

or a pharmaceutically acceptable salt thereof, wherein: each E is independently selected from the group consisting of a bond, CR⁵R⁶, O, C(O), and NR¹⁰; R¹⁰ is selected from the group consisting of H, C_1-6 alkyl, C_1-6 haloalkyl, and C_1-6 alkylcarbonyl; t and u are each independently 0, 1 , or 2; and is a single bond, a cis double bond, or a trans double bond.

In an embodiment, Ring A and Ring B are each independently selected from the group consisting of absent, phenyl, 5- or 6-membered heterocycloalkyl, 5- or 6-membered heteroaryl, and 8- or 9-membered bicyclic ring.

In another embodiment, Ring A and Ring B are each independently selected from the group consisting of absent, phenyl, 6-membered heterocycloalkyl, 5- or 6-membered heteroaryl, and 9-membered bicyclic ring.

In yet another embodiment, the heterocycloalkyl, heteroaryl, and bicyclic ring of Ring A and Ring B contains 1 , 2, or 3 heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur.

In still another embodiment, the heterocycloalkyl, heteroaryl, and bicyclic ring of Ring A and Ring B contains 1 or 2 heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur.

In an embodiment, Ring A is selected from the group consisting of absent, phenyl, 5- or 6-membered heteroaryl, and 9-membered bicyclic ring.

In another embodiment, Ring B is selected from the group consisting of absent, 6- membered heterocycloalkyl, and 5-membered heteroaryl.

In yet another embodiment, one of Ring A or Ring B is not absent.

In still another embodiment, the compound of Formula (IA-II) is a compound of Formula (lA-lla):

or a pharmaceutically acceptable salt thereof, wherein:

X³ is N or CH, provided that when E is bonded to X³, then X³ is C; each E is independently selected from the group consisting of a bond, CR⁵R⁶, O, C(O), and NR¹⁰; R¹⁰ is selected from the group consisting of H, C_1-6 alkyl, C_1-6 haloalkyl, and C_1-6 alkylcarbonyl; t and u are each independently 0, 1 , or 2; and is a single bond, a cis double bond, or a trans double bond.

In an embodiment, the compound of Formula (IA-II) is a compound of Formula (IA-

or a pharmaceutically acceptable salt thereof, wherein: each X⁴ is independently N or CH; each E is independently selected from the group consisting of a bond, CR⁵R⁶, O, C(O), and NR¹⁰;

R¹⁰ is selected from the group consisting of H, C_1-6 alkyl, C_1-6 haloalkyl, and C_1-6 alkylcarbonyl; t and u are each independently 0, 1 , or 2; and is a single bond, a cis double bond, or a trans double bond.

In another embodiment, the compound of Formula (IA-II) is a compound of Formula

(lA-llc):

or a pharmaceutically acceptable salt thereof, wherein: each X⁵ is independently NR¹¹ or CH₂, provided that when E is bonded to an X⁵, then said X⁵ is N or CH; each E is independently selected from the group consisting of a bond, CR⁵R⁶, O, C(O), and NR¹⁰;

R¹¹ is selected from the group consisting of C_1-6 alkyl, C_1-6 alkoxy, C_1-6 haloalkyl, and C_1-6 alkylcarbonyl; t and u are each independently 0, 1 , or 2; and is a single bond, a cis double bond, or a trans double bond.

In yet another embodiment, the compound of Formula (lA-lla) is a compound of Formula (lA-llaa):

or a pharmaceutically acceptable salt thereof, wherein: each E is independently selected from the group consisting of a bond, CR⁵R⁶, O, C(O), and NR¹⁰;

In still another embodiment, the compound of Formula (lA-llb) is a compound of Formula (lA-llba):

In an embodiment, the compound of Formula (lA-llc) is a compound of Formula (IA-

Ilea):

In another embodiment, the compound of Formula (lA-llc) is a compound of Formula (lA-llcb):

In yet another embodiment, R¹ is C_1-6 alkyl.

In still another embodiment, R³ and R⁴ are each independently selected from the group consisting of C_1-6 alkyl, C_1-6 alkoxy, C_1-6 haloalkyl, and C_1-6 alkylcarbonyl, wherein C_1-6 alkoxy is optionally substituted with one R⁹.

In an embodiment, R³ is selected from the group consisting of C_1-6 alkyl, C_1-6 alkoxy, and C_1-6 alkylcarbonyl, wherein C_1-6 alkoxy is optionally substituted with one R⁹.

In another embodiment, R⁵ and R⁶ are each independently selected from the group consisting of H, halo, C_1-6 alkyl, and C_1-6 haloalkyl.

In yet another embodiment, L₃ is a bond.

In still another embodiment, n is 1.

In an embodiment, p is 0 or 1 .

In another embodiment, r is 0 or 1 .

In yet another embodiment, Y is O.

In still another embodiment, at least one E is a bond.

In an embodiment, at least one E is O. In another embodiment, at least two E are either O or NR¹⁰.

In yet another embodiment, at least two E are a bond.

In still another embodiment, at least three E are a bond.

In an embodiment, the compound of Formula I is selected from the group consisting of a compound in Table 1 , or a pharmaceutically acceptable salt thereof.

Table 1

The compounds disclosed herein may exist as tautomers and optical isomers (e.g., enantiomers, diastereomers, diastereomeric mixtures, racemic mixtures, and the like).

It is generally well known in the art that any compound that will be converted in vivo to provide a compound disclosed herein is a prodrug within the scope of the present disclosure.

Compounds provided herein can also include all isotopes of atoms occurring in the intermediates or final compounds. Isotopes include those atoms having the same atomic number but different mass numbers. For example, isotopes of hydrogen include tritium and deuterium. One or more constituent atoms of the compounds of the invention can be replaced or substituted with isotopes of the atoms in natural or non-natural abundance. In some embodiments, the compound includes at least one deuterium atom. For example, one or more hydrogen atoms in a compound of the present disclosure can be replaced or substituted by deuterium. In some embodiments, the compound includes two or more deuterium atoms. In some embodiments, the compound includes 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 deuterium atoms. Synthetic methods for including isotopes into organic compounds are known in the art (Deuterium Labeling in Organic Chemistry by Alan F. Thomas (New York, N.Y., Appleton-Century-Crofts, 1971 ; The Renaissance of H/D Exchange by Jens Atzrodt, Volker Derdau, Thorsten Fey and Jochen Zimmermann, Angew. Chem. Int. Ed. 2007, 7744-7765; The Organic Chemistry of Isotopic Labelling by James R. Hanson, Royal Society of Chemistry, 2011 ). Isotopically labeled compounds can used in various studies such as NMR spectroscopy, metabolism experiments, and/or assays.

In an aspect, provided herein is a pharmaceutical composition comprising any one of the compounds disclosed herein, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier.

In an embodiment, the composition further comprises a second active agent. In another embodiment, the second active agent is selected from the group consisting of a MEK inhibitor, a PI3K inhibitor, and an mTor inhibitor. In yet another embodiment, the second active agent prevents EGFR dimer formation in a subject. In still another embodiment, the second active agent is selected from the group consisting of cetuximab, trastuzumab, and panitumumab. In an embodiment, the second active agent is an ATP competitive EGFR inhibitor. In another embodiment, the ATP competitive EGFR inhibitor is osimertinib, gefitinib, or erlotinib. In another embodiment, the ATP competitive EGFR inhibitor is osimertinib.

In another aspect, provided herein are pharmaceutical compositions comprising a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

In another aspect, provided herein is a method of inhibiting the activity of EGFR, comprising administering to a subject in need thereof an effective amount of a compound of disclosed herein or a pharmaceutical composition comprising a compound disclosed herein and a pharmaceutically acceptable carrier. In an embodiment, the compound targets Cys775 on EGFR.

In another aspect, the pharmaceutical composition further comprises a second active agent, wherein said second active agent prevents EGFR dimer formation, and a pharmaceutically acceptable carrier. In some embodiments, the second active agent that prevents EGFR dimer formation is an antibody. In further embodiments, the second active agent that prevents EGFR dimer formation is cetuximab, trastuzumab, or panitumumab. In further embodiments, the second active agent that prevents EGFR dimer formation is cetuximab.

A compound that binds to an allosteric site in EGFR, such as the compounds of the present disclosure (e.g., the compounds of the formulae disclosed herein), optionally in combination with a second active agent, wherein said second active agent prevents EGFR dimer formation, are capable of modulating EGFR activity. In some embodiments, the compounds of the present disclosure are capable of inhibiting or decreasing EGFR activity without a second active agent (e.g., an antibody such as cetuximab, trastuzumab, or panitumumab). In other embodiments, the compounds of the present disclosure in combination with a second active agent. In an embodiment, the second active agent prevents EGFR dimer formation and/or are capable of inhibiting or decreasing EGFR activity. In some embodiments, the second active agent that prevents EGFR dimer formation is an antibody. In further embodiments, the second active agent that prevents EGFR dimer formation is cetuximab, trastuzumab, or panitumumab. In further embodiments, the second active agent that prevents EGFR dimer formation is cetuximab. In an embodiment, the second active agent is an ATP competitive EGFR inhibitor. In another embodiment, the ATP competitive EGFR inhibitor is osimertinib, gefitinib or erlotinib. In another embodiment, the ATP competitive EGFR inhibitor is osimertinib.

Methods of Treatment

In an aspect, provided herein is a method of treating cancer in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of a compound disclosed herein, or a pharmaceutically acceptable salt thereof. In an embodiment, the cancer is selected from the group consisting of lung cancer, colon cancer, breast cancer, endometrial cancer, thyroid cancer, glioma, squamous cell carcinoma, and prostate cancer. In another embodiment, the cancer is non-small cell lung cancer (NSCLC).

In another aspect, provided herein is a method of inhibiting the activity of EGFR in an subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound provided herein, or a pharmaceutically acceptable salt thereof.

In yet another aspect, provided herein is a method of inhibiting a kinase in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of a compound provided herein. In an embodiment, the kinase is EGFR. In another embodiment, the kinase is HER.

In yet another aspect, provided herein is a method of treating or preventing a kinase- mediated disorder in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of a compound of the present disclosure. In an embodiment, the kinase-mediated disorder is resistant to an EGFR-targeted therapy. In another embodiment, the EGFR-treated therapy is selected from the group consisting of gefitinib, erlotinib, osimertinib, CO-1686, and WZ4002.

In still another aspect, provided herein is a method of inhibiting the activity of EGFR in a subject in need thereof comprising targeting both Cys775 and Cys797 on EGFR. In yet another aspect, provided herein is a method of inhibiting the activity of EGFR in a subject in need thereof comprising administering a compound that targets both Cys775 and Cys797 on EGFR. The compound can simultaneously form two covalent bonds to cysteine 797 and cysteine 775. In an embodiment, the compound is a compound of Formula I, described herein. In some embodiments, the compounds of the present disclosure are capable of modulating (e.g., inhibiting or decreasing) the activity of EGFR containing one or more mutations. In some embodiments, the mutant EGFR contains one or more mutations selected from T790M, L718Q, L844V, V948R, L858R, 1941 R, C797S, and Del. In other embodiments, the mutant EGFR contains a combination of mutations, wherein the combination is selected from Del/L718Q, Del/L844V, Del/T790M, Del/T790M/L718Q, Del/T790M/L844V, L858R/L718Q, L858R/L844V, L858R/T790M, L858R/T790M/I941R, Del/T790M, Del/T790M/C797S, L858R/T790M/C797S, and L858R/T790M/L718Q. In other embodiments, the mutant EGFR contains a combination of mutations, wherein the combination is selected from Del/L844V, L858R/L844V, L858R/T790M, L858R/T790M/I941R, L858R/T790M/C797S, Del/T790M, Del/T790M, Del/T790M/C797S, and L858R/T790M. In other embodiments, the mutant EGFR contains a combination of mutations, wherein the combination is selected from L858R/T790M, L858R/T790M/I941R, L858R/T790M/C797S, Del/T790M, Del/T790M/C797S, and L858R/T790M.

In some embodiments, the compounds of the present disclosure in combination with a second active agent, wherein said second active agent prevents EGFR dimer formation, are capable of modulating (e.g., inhibiting or decreasing) the activity of EGFR containing one or more mutations. In some embodiments, the second active agent that prevents EGFR dimer formation is an antibody. In further embodiments, the second active agent that prevents EGFR dimer formation is cetuximab, trastuzumab, or panitumumab. In further embodiments, the second active agent that prevents EGFR dimer formation is cetuximab. In an embodiment, the second active agent is an ATP competitive EGFR inhibitor. In another embodiment, the ATP competitive EGFR inhibitor is osimertinib, gefitinib or erlotinib.

In some embodiments, the compounds of the present disclosure are capable of modulating (e.g., inhibiting or decreasing) the activity of EGFR containing one or more mutations, but do not affect the activity of a wild-type EGFR.

Modulation of EGFR containing one or more mutations, such as those described herein, but not a wild-type EGFR, provides an approach to the treatment, prevention, or amelioration of diseases including, but not limited to, cancer and metastasis, inflammation, arthritis, systemic lupus erythematosus, skin-related disorders, pulmonary disorders, cardiovascular disease, ischemia, neurodegenerative disorders, liver disease, gastrointestinal disorders, viral and bacterial infections, central nervous system disorders, Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, spinal cord injury, and peripheral neuropathy.

In some embodiments, the compounds of the disclosure exhibit greater inhibition of EGFR containing one or more mutations as described herein relative to a wild-type EGFR. In certain embodiments, the compounds of the disclosure exhibit at least 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold or 100-fold greater inhibition of EGFR containing one or more mutations as described herein relative to a wild-type EGFR. In various embodiments, the compounds of the disclosure exhibit up to 1000-fold greater inhibition of EGFR containing one or more mutations as described herein relative to a wild-type EGFR. In various embodiments, the compounds of the disclosure exhibit up to 10000-fold greater inhibition of EGFR having a combination of mutations described herein (e.g., L858R/T790M, L858R/T790M/I941R, L858R/T790M/C797S, Del/T790M, Del/T790M/C797S, and L858R/T790M) relative to a wild-type EGFR.

In some embodiments, the inhibition of EGFR activity is measured by IC₅₀. A compound with a lower IC₅₀ value, as determined under substantially similar conditions, is a more potent inhibitor relative to a compound with a higher IC₅₀ value. In some embodiments, the substantially similar conditions comprise determining an EGFR-dependent phosphorylation level, in vitro or in vivo (e.g., in 3T3 cells expressing a wild type EGFR, a mutant EGFR, or a fragment of any thereof).

In some embodiments, the inhibition of EGFR activity is measured by EC₅₀. A compound with a lower EC₅₀ value, as determined under substantially similar conditions, is a more potent inhibitor relative to a compound with a higher EC₅₀ value. In some embodiments, the substantially similar conditions comprise determining an EGFR-dependent phosphorylation level, in vitro or in vivo (e.g., in 3T3 cells expressing a wild type EGFR, a mutant EGFR, or a fragment of any thereof).

In some embodiments, the inhibition of EGFR by a compound of the disclosure can be measured via a biochemical assay. By illustrative and non-limiting example, a homogenous time-resolved fluorescence (HTRF) assay may be used to determine inhibition of EGFR activity using conditions and experimental parameters disclosed herein. The HTRF assay may, for example, employ concentrations of substrate (e.g., biotin-Lck-peptide substrate) of about 1 μM; concentrations of EGFR (mutant or WT) from about 0.2 nM to about 40 nM; and concentrations of inhibitor from about 0.000282 μM to about 50 μM. A compound of the disclosure screened under these conditions may, for example, exhibit an IC₅₀ value from about 1 nM to >1 μM; from about 1 nM to about 400 nM; from about 1 nM to about 150 nM; from about 1 nM to about 75 nM; from about 1 nM to about 40 nM; from about 1 nM to about 25 nM; from about 1 nM to about 15 nM; or from about 1 nM to about 10 nM. In certain embodiments, a compound of the disclosure screened under the above conditions for inhibition of EGFR having a mutation or combination of mutations selected from L858R/T790M, L858R, and T790M may, for example, exhibit an IC₅₀ value from about 1 nM to >1 μM; from about 1 nM to about 400 nM; from about 1 nM to about 150 nM; from about 1 nM to about 75 nM; from about 1 nM to about 40 nM; from about 1 nM to about 25 nM; from about 1 nM to about 15 nM; or from about 1 nM to about 10 nM. In some embodiments, the compounds of the disclosure bind to an allosteric site in EGFR. In some embodiments, the compounds of the disclosure interact with at least one amino acid residue of epidermal growth factor receptor (EGFR) selected from Lys745, Leu788, and Ala 743. In other embodiments, the compounds of the disclosure interact with at least one amino acid residue of epidermal growth factor receptor (EGFR) selected from Cys755, Leu777, Phe856, and Asp855. In other embodiments, the compounds of the disclosure interact with at least one amino acid residue of epidermal growth factor receptor (EGFR) selected from Met766, Ile759, Glu762, and Ala763. In other embodiments, the compounds of the disclosure interact with at least one amino acid residue of epidermal growth factor receptor (EGFR) selected from Lys745, Leu788, and Ala 743; at least one amino acid residue of epidermal growth factor receptor (EGFR) selected from Cys755, Leu777, Phe856, and Asp855; and at least one amino acid residue of epidermal growth factor receptor (EGFR) selected from Met766, IIe759, Glu762, and Ala763. In other embodiments, the compounds of the disclosure do not interact with any of the amino acid residues of epidermal growth factor receptor (EGFR) selected from Met793, Gly796, and Cys797.

An EGFR sensitizing mutation comprises without limitation L858R, G719S, G719C, G719A, L861Q, a deletion in exon 19 and/or an insertion in exon 20. A drug-resistant EGFR mutant can have without limitation a drug resistance mutation comprising T790M, T854A, L718Q, C797S, or D761Y.

The selectivity between wild-type EGFR and EGFR containing one or more mutations as described herein can also be measured using cellular proliferation assays where cell proliferation is dependent on kinase activity. For example, murine Ba/F3 cells transfected with a suitable version of wild-type EGFR (such as VIII; containing a WT EGFR kinase domain), or Ba/F3 cells transfected with L858R/T790M, Del/T790M/L718Q, L858R/T790M/L718Q, L858R/T790M/C797S, Del/T790M/C797S, L858R/T790M/I941R, or Exon 19 deletion/T790M can be used. Proliferation assays are performed at a range of inhibitor concentrations (10 μM, 3 μM, 1.1 μM, 330 nM, 110 nM, 33 nM, 11 nM, 3 nM, 1 nM) and an EC₅₀ is calculated.

In still another aspect, the disclosure provides a method of inhibiting epidermal growth factor receptor (EGFR), the method comprising administering to a subject in need thereof an effective amount of a compound disclosed herein, or a pharmaceutically acceptable salt thereof. In some embodiments, the method further comprises administering a second active agent, wherein said second active agent prevents EGFR dimer formation. In some embodiments, the second active agent that prevents EGFR dimer formation is an antibody. In further embodiments, the second active agent that prevents EGFR dimer formation is cetuximab, trastuzumab, or panitumumab. In further embodiments, the second active agent that prevents EGFR dimer formation is cetuximab. In an embodiment, the second active agent is an ATP competitive EGFR inhibitor. In another embodiment, the ATP competitive EGFR inhibitor is osimertinib, gefitinib or erlotinib. In another embodiment, the ATP competitive EGFR inhibitor is osimertinib.

In another aspect, provided herein is a method of treating or preventing a disease, the method comprising administering to a subject in need thereof an effective amount of a compound disclosed herein, or a pharmaceutically acceptable salt thereof. In some embodiments, the disease is mediated by a kinase. In further embodiments, the kinase comprises a mutated cysteine residue. In further embodiments, the mutated cysteine residue is located in or near the position equivalent to Cys 797 in EGFR, including such positions in Jak3, Blk, Bmx, Btk, HER2 (ErbB2), HER4 (ErbB4), Itk, Tec, and Txk. In some embodiments, the method further comprises administering a second active agent, wherein said second active agent prevents dimer formation of the kinase. In some embodiments, the second active agent that prevents kinase dimer formation is an antibody. In further embodiments, the second active agent prevents EGFR dimer formation. In further embodiments, the second active agent that prevents EGFR dimer formation is cetuximab, trastuzumab, or panitumumab. In further embodiments, the second active agent that prevents EGFR dimer formation is cetuximab. In an embodiment, the second active agent is an ATP competitive EGFR inhibitor. In another embodiment, the ATP competitive EGFR inhibitor is osimertinib, gefitinib or erlotinib. In another embodiment, the ATP competitive EGFR inhibitor is osimertinib.

In some embodiments, the disease is mediated by EGFR (e.g., EGFR plays a role in the initiation or development of the disease). In some embodiments, the disease is mediated by a Her-kinase. In further embodiments, the Her-kinase is HER1 , HER2, or HER4.

In certain embodiments, the disease is resistant to a known EGFR inhibitor, including but not limited to, gefitinib, erlotinib, osimertinib, CO-1686, or WZ4002. In certain embodiments, a diagnostic test is performed to determine if the disease is associated with an activating mutation in EGFR. In certain embodiments, a diagnostic test is performed to determine if the disease is associated with an EGFR harboring an activating mutation and/or a drug resistance mutation. Activating mutations comprise without limitation L858R, G719S, G719C, G719A, L718Q, L861Q, a deletion in exon 19 and/or an insertion in exon 20. Drug resistant EGFR mutants can have without limitation a drug resistance mutation comprising T790M, T854A, L718Q, C797S, or D761Y. The diagnostic test can comprise sequencing, pyrosequencing, PCR, RT-PCR, or similar analysis techniques known to those of skill in the art that can detect nucleotide sequences.

In certain embodiments, the disease is cancer or a proliferation disease. In further embodiments, the disease is lung cancer, colon cancer, breast cancer, prostate cancer, liver cancer, pancreas cancer, brain cancer, kidney cancer, ovarian cancer, stomach cancer, skin cancer, bone cancer, gastric cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, hepatocellular carcinoma, papillary renal carcinoma, head and neck squamous cell carcinoma, leukemias, lymphomas, myelomas, or solid tumors. In further embodiments, the disease is lung cancer, breast cancer, glioma, squamous cell carcinoma, or prostate cancer. In still further embodiments, the disease is non-small cell lung cancer.

In yet another aspect, provided herein is a method of treating a kinase-mediated disorder comprising administering to a subject in need thereof an effective amount of a compound disclosed herein, or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is an inhibitor of HER1 , HER2, or HER4. In other embodiments, the subject is administered an additional therapeutic agent. In other embodiments, the compound and the additional therapeutic agent are administered simultaneously or sequentially.

In another aspect, the disclosure provides a method of treating a kinase mediated disorder, the method comprising administering to a subject in need thereof an effective amount of a compound disclosed herein, or a pharmaceutically acceptable salt thereof, and a second active agent, wherein said second active agent prevents EGFR dimer formation. In some embodiments, the compound is an inhibitor of HER1 , HER2, or HER4. In other embodiments, the subject is administered an additional therapeutic agent. In other embodiments, the compound, the second active agent that prevents EGFR dimer formation, and the additional therapeutic agent are administered simultaneously or sequentially. In some embodiments, the second active agent that prevents EGFR dimer formation is an antibody. In further embodiments, the second active agent that prevents EGFR dimer formation is cetuximab, trastuzumab, or panitumumab. In further embodiments, the second active agent that prevents EGFR dimer formation is cetuximab. In an embodiment, the second active agent is an ATP competitive EGFR inhibitor. In another embodiment, the ATP competitive EGFR inhibitor is osimertinib, gefitinib or erlotinib. In another embodiment, the ATP competitive EGFR inhibitor is osimertinib.

In other embodiments, the disease is cancer. In further embodiments, the cancer is lung cancer, colon cancer, breast cancer, prostate cancer, liver cancer, pancreas cancer, brain cancer, kidney cancer, ovarian cancer, stomach cancer, skin cancer, bone cancer, gastric cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, hepatocellular carcinoma, papillary renal carcinoma, head and neck squamous cell carcinoma, leukemias, lymphomas, myelomas, or solid tumors. In further embodiments, the disease is lung cancer, breast cancer, glioma, squamous cell carcinoma, or prostate cancer. In still further embodiments, the disease is non-small cell lung cancer.

In certain embodiments, the EGFR activation is selected from mutation of EGFR, amplification of EGFR, expression of EGFR, and ligand mediated activation of EGFR.

In further embodiments, the mutation of EGFR is selected from G719S, G719C, G719A, L858R, L861Q, an exon 19 deletion mutation, and an exon 20 insertion mutation.

In still another aspect, provided herein is a method of treating cancer in a subject, wherein the subject is identified as being in need of EGFR inhibition for the treatment of cancer, comprising administering to the subject an effective amount of a compound disclosed herein, or a pharmaceutically acceptable salt thereof.

In certain embodiments, the subject identified as being in need of EGFR inhibition is resistant to a known EGFR inhibitor, including but not limited to, gefitinib, erlotinib, osimertinib, CO-1686, or WZ4002. In certain embodiments, a diagnostic test is performed to determine if the subject has an activating mutation in EGFR. In certain embodiments, a diagnostic test is performed to determine if the subject has an EGFR harboring an activating mutation and/or a drug resistance mutation. Activating mutations comprise without limitation L858R, G719S, G719C, G719A, L718Q, L861Q, a deletion in exon 19 and/or an insertion in exon 20. Drug resistant EGFR mutants can have without limitation a drug resistance mutation comprising T790M, T854A, L718Q, C797S, or D761Y. The diagnostic test can comprise sequencing, pyrosequencing, PCR, RT-PCR, or similar analysis techniques known to those of skill in the art that can detect nucleotide sequences.

In an aspect, provided herein is a method of preventing resistance to a known EGFR inhibitor (including but not limited to gefitinib, erlotinib, osimertinib, CO-1686, or WZ4002) in a subject, comprising administering to a subject in need thereof an effective amount of a compound disclosed herein, or a pharmaceutically acceptable salt thereof.

In another aspect, provided herein is a method of preventing resistance to a known EGFR inhibitor (including but not limited to gefitinib, erlotinib, osimertinib, CO-1686, or WZ4002) in a disease, comprising administering to a subject in need thereof an effective amount of a compound disclosed herein, or a pharmaceutically acceptable salt thereof, and a second active agent, wherein said second active agent prevents EGFR dimer formation. In some embodiments, the second active agent that prevents EGFR dimer formation is an antibody. In further embodiments, the second active agent that prevents EGFR dimer formation is cetuximab, trastuzumab, or panitumumab. In further embodiments, the second active agent that prevents EGFR dimer formation is cetuximab.

In an embodiment of the methods disclosed herein, the subject is a human.

In another aspect, the disclosure provides a compound disclosed herein, or a pharmaceutically acceptable salt thereof, for use in the manufacture of a medicament for treating or preventing a disease in which EGFR plays a role.

In an aspect, provided herein is a method of treating or preventing a condition selected from the group consisting of autoimmune diseases, inflammatory diseases, proliferative and hyperproliferative diseases, immunologically-mediated diseases, bone diseases, metabolic diseases, neurological and neurodegenerative diseases, cardiovascular diseases, hormone related diseases, allergies, asthma, and Alzheimer's disease. In other embodiments, said condition is selected from a proliferative disorder and a neurodegenerative disorder.

One aspect of this disclosure provides compounds that are useful for the treatment of diseases, disorders, and conditions characterized by excessive or abnormal cell proliferation. Such diseases include, but are not limited to, a proliferative or hyperproliferative disease, and a neurodegenerative disease. Examples of proliferative and hyperproliferative diseases include, without limitation, cancer.

In one aspect, the present disclosure provides for the use of one or more compounds of the disclosure in the manufacture of a medicament for the treatment of cancer, including without limitation the various types of cancer disclosed herein.

In some embodiments, the compounds of this disclosure are useful for treating cancer, such as colorectal, thyroid, breast, and lung cancer; and myeloproliferative disorders, such as polycythemia vera, thrombocythemia, myeloid metaplasia with myelofibrosis, chronic myelogenous leukemia, chronic myelomonocytic leukemia, hypereosinophilic syndrome, juvenile myelomonocytic leukemia, and systemic mast cell disease. In some embodiments, the compounds of this disclosure are useful for treating hematopoietic disorders, in particular, acute-myelogenous leukemia (AML), chronic- myelogenous leukemia (CML), acute-promyelocytic leukemia, and acute lymphocytic leukemia (ALL).

The disclosure further provides a method for the treatment or prevention of cell proliferative disorders such as hyperplasias, dysplasias and pre-cancerous lesions. Dysplasia is the earliest form of pre-cancerous lesion recognizable in a biopsy by a pathologist. The subject compounds may be administered for the purpose of preventing said hyperplasias, dysplasias, or pre-cancerous lesions from continuing to expand or from becoming cancerous. Examples of pre-cancerous lesions may occur in skin, esophageal tissue, breast and cervical intra-epithelial tissue.

Examples of neurodegenerative diseases include, without limitation, adrenoleukodystrophy (ALD), Alexander's disease, Alper's disease, Alzheimer's disease, amyotrophic lateral sclerosis (Lou Gehrig's Disease), ataxia telangiectasia, Batten disease (also known as Spielmeyer-Vogt-Sjogren-Batten disease), bovine spongiform encephalopathy (BSE), Canavan disease, Cockayne syndrome, corticobasal degeneration, Creutzfeldt-Jakob disease, familial fatal insomnia, frontotemporal lobar degeneration, Huntington's disease, HIV-associated dementia, Kennedy's disease, Krabbe's disease, Lewy body dementia, neuroborreliosis, Machado-Joseph disease (spinocerebellar ataxia type 3), multiple system atrophy, multiple sclerosis, narcolepsy, Niemann Pick disease, Parkinson's disease, Pelizaeus-Merzbacher disease, Pick's disease, primary lateral sclerosis, prion diseases, progressive supranuclear palsy, Refsum's disease, Sandhoff disease, Schilder's disease, subacute combined degeneration of spinal cord secondary to pernicious anaemia, Spielmeyer-Vogt-Sjogren-Batten disease (also known as Batten disease), spinocerebellar ataxia (multiple types with varying characteristics), spinal muscular atrophy, Steele- Richardson-Olszewski disease, tabes dorsalis, and toxic encephalopathy.

Another aspect of this disclosure provides a method for the treatment or lessening the severity of a disease selected from a proliferative or hyperproliferative disease, or a neurodegenerative disease, comprising administering an effective amount of a compound, or a pharmaceutically acceptable composition comprising a compound, to a subject in need thereof. In other embodiments, the method further comprises administering a second active agent, wherein said second active agent prevents EGFR dimer formation. In some embodiments, the second active agent that prevents EGFR dimer formation is an antibody. In further embodiments, the second active agent that prevents EGFR dimer formation is cetuximab, trastuzumab, or panitumumab. In further embodiments, the second active agent that prevents EGFR dimer formation is cetuximab. In an embodiment, the second active agent is an ATP competitive EGFR inhibitor. In another embodiment, the ATP competitive EGFR inhibitor is osimertinib, gefitinib or erlotinib. In another embodiment, the ATP competitive EGFR inhibitor is osimertinib.

The activity of the compounds and compositions of the present disclosure as EGFR kinase inhibitors may be assayed in vitro, in vivo, or in a cell line. In vitro assays include assays that determine inhibition of either the kinase activity or ATPase activity of the activated kinase. Alternate in vitro assays quantitate the ability of the inhibitor to bind to the protein kinase and may be measured either by radio labelling the inhibitor prior to binding, isolating the inhibitor/kinase complex and determining the amount of radio label bound, or by running a competition experiment where new inhibitors are incubated with the kinase bound to known radioligands. Detailed conditions for assaying a compound utilized in this disclosure as an inhibitor of various kinases are set forth in the Examples below.

In accordance with the foregoing, the present disclosure further provides a method for preventing or treating any of the diseases or disorders described above in a subject in need of such treatment, which method comprises administering to said subject a therapeutically effective amount of a compound of the disclosure, or a pharmaceutically acceptable salt thereof, and optionally a second active agent, wherein said second active agent prevents EGFR dimer formation. For any of the above uses, the required dosage will vary depending on the mode of administration, the particular condition to be treated and the effect desired.

In other embodiments, the compound and the second active agent that prevents EGFR dimer formation are administered simultaneously or sequentially.

Administration / Dosages / Formulations

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butylene glycol, dimethylformamide, oils (in particular, cottonseed, groundnut, com, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

Injectable preparations (for example, sterile injectable aqueous or oleaginous suspensions) may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution, suspension, or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1 ,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, U.S.P., and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables. In order to prolong the effect of a drug, it is often desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the drug then depends upon its rate of dissolution which, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle.

Compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing the compounds of this disclosure with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol, or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.

Solid compositions of a similar type may also be employed as fillers in soft and hard filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.

The active compounds can also be in micro-encapsulated form with one or more excipients as noted above. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings, and other coatings well known in the pharmaceutical formulating art. In such solid dosage forms the active compound may be admixed with at least one inert diluent such as sucrose, lactose or starch. Such dosage forms may also comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such a magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets, and pills, the dosage forms may also comprise buffering agents.

Dosage forms for topical or transdermal administration of a compound of this disclosure include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The active component is admixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives or buffers as may be required. Ophthalmic formulation, ear drops, eye ointments, powders and solutions are also contemplated as being within the scope of this disclosure.

The ointments, pastes, creams and gels may contain, in addition to an active compound of this disclosure, excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to the compounds of this disclosure, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays can additionally contain customary propellants such as chlorofluorohydrocarbons.

Transdermal patches have the added advantage of providing controlled delivery of a compound to the body. Such dosage forms can be made by dissolving or dispensing the compound in the proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by either providing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel.

According to the methods of treatment of the present disclosure, disorders are treated or prevented in a subject, such as a human or other animal, by administering to the subject a therapeutically effective amount of a compound of the disclosure, in such amounts and for such time as is necessary to achieve the desired result. The term “therapeutically effective amount” of a compound of the disclosure, as used herein, means a sufficient amount of the compound so as to decrease the symptoms of a disorder in a subject. As is well understood in the medical arts a therapeutically effective amount of a compound of this disclosure will be at a reasonable benefit/risk ratio applicable to any medical treatment.

In general, compounds of the disclosure will be administered in therapeutically effective amounts via any of the usual and acceptable modes known in the art, either singly or in combination with one or more therapeutic agents. A therapeutically effective amount may vary widely depending on the severity of the disease, the age and relative health of the subject, the potency of the compound used and other factors. In general, satisfactory results are indicated to be obtained systemically at daily dosages of from about 0.03 to 2.5 mg/kg per body weight. An indicated daily dosage in the larger mammal, e.g., humans, is in the range from about 0.5 mg to about 100 mg, conveniently administered, e.g., in divided doses up to four times a day or in retard form. Suitable unit dosage forms for oral administration comprise from ca. 1 to 50 mg active ingredient.

In certain embodiments, a therapeutic amount or dose of the compounds of the present disclosure may range from about 0.1 mg/Kg to about 500 mg/Kg, alternatively from about 1 to about 50 mg/Kg. In general, treatment regimens according to the present disclosure comprise administration to a patient in need of such treatment from about 10 mg to about 1000 mg of the compound(s) of this disclosure per day in single or multiple doses. Therapeutic amounts or doses will also vary depending on route of administration, as well as the possibility of co-usage with other agents.

Upon improvement of a subject's condition, a maintenance dose of a compound, composition or combination of this disclosure may be administered, if necessary. Subsequently, the dosage or frequency of administration, or both, may be reduced, as a function of the symptoms, to a level at which the improved condition is retained; when the symptoms have been alleviated to the desired level, treatment should cease. The subject may, however, require intermittent treatment on a long-term basis upon any recurrence of disease symptoms.

It will be understood, however, that the total daily usage of the compounds and compositions of the present disclosure will be decided by the attending physician within the scope of sound medical judgment. The specific inhibitory dose for any particular patient will depend upon a variety of factors including the disorder being treated and the severity of the disorder; the activity of the specific compound employed; the specific composition employed; the age, body weight, general health, sex and diet of the patient; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific compound employed; and like factors well known in the medical arts.

The disclosure also provides for a pharmaceutical combination, e.g., a kit, comprising a) a first agent which is a compound of the disclosure as disclosed herein, in free form or in pharmaceutically acceptable salt form, and b) at least one co-agent. The kit can comprise instructions for its administration.

In certain embodiments, these compositions optionally further comprise one or more additional therapeutic agents. For example, an agent that prevents EGFR dimer formation, chemotherapeutic agents or other antiproliferative agents may be combined with the compounds of this disclosure to treat proliferative diseases and cancer.

Some examples of materials which can serve as pharmaceutically acceptable carriers include, but are not limited to, ion exchangers; alumina; aluminum stearate; lecithin; serum proteins, such as human serum albumin; buffer substances such as phosphates, glycine, sorbic acid, or potassium sorbate; partial glyceride mixtures of saturated vegetable fatty acids; water; salts or electrolytes, such as protamine sulfate; disodium hydrogen phosphate; potassium hydrogen phosphate; sodium chloride; zinc salts; colloidal silica; magnesium trisilicate; polyvinyl pyrrolidone; polyacrylates; waxes; polyethylenepolyoxypropylene-block polymers; wool fat; sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols, such a propylene glycol or polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; and phosphate buffer solutions. Further, non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the composition, according to the judgment of the formulator. The protein kinase inhibitors or pharmaceutical salts thereof may be formulated into pharmaceutical compositions for administration to animals or humans. These pharmaceutical compositions, which comprise an amount of the protein inhibitor effective to treat or prevent a protein kinase-mediated condition and a pharmaceutically acceptable carrier, are other embodiments of the present disclosure.

Kits

In an aspect, provided herein is a kit comprising a compound capable of inhibiting kinase activity selected from one or more compounds of disclosed herein, or pharmaceutically acceptable salts thereof, and instructions for use in treating cancer. In certain embodiments, the kit further comprises components for performing a test to determine whether a subject has activating and/or drug resistance mutations in EGFR.

In another aspect, the disclosure provides a kit comprising a compound capable of inhibiting EGFR activity selected from a compound disclosed herein, or a pharmaceutically acceptable salt thereof.

In another aspect, the disclosure provides a kit comprising a compound capable of inhibiting kinase activity selected from one or more compounds of disclosed herein, or pharmaceutically acceptable salts thereof; a second active agent, wherein said second active agent prevents EGFR dimer formation; and instructions for use in treating cancer. In certain embodiments, the kit further comprises components for performing a test to determine whether a subject has activating and/or drug resistance mutations in EGFR. In some embodiments, the second active agent that prevents EGFR dimer formation is an antibody. In further embodiments, the second active agent that prevents EGFR dimer formation is cetuximab, trastuzumab, or panitumumab. In further embodiments, the second active agent that prevents EGFR dimer formation is cetuximab.

In another aspect, the disclosure provides a kit comprising a compound capable of inhibiting EGFR activity selected from a compound of disclosed herein, or a pharmaceutically acceptable salt thereof and a second active agent, wherein said second active agent prevents EGFR dimer formation. In some embodiments, the second active agent that prevents EGFR dimer formation is an antibody. In further embodiments, the second active agent that prevents EGFR dimer formation is cetuximab, trastuzumab, or panitumumab. In further embodiments, the second active agent that prevents EGFR dimer formation is cetuximab. In an embodiment, the second active agent is an ATP competitive EGFR inhibitor. In another embodiment, the ATP competitive EGFR inhibitor is osimertinib, gefitinib or erlotinib. In another embodiment, the ATP competitive EGFR inhibitor is osimertinib. The disclosure is further illustrated by the following examples and synthesis schemes, which are not to be construed as limiting this disclosure in scope or spirit to the specific procedures herein described. It is to be understood that the examples are provided to illustrate certain embodiments and that no limitation to the scope of the disclosure is intended thereby. It is to be further understood that resort may be had to various other embodiments, modifications, and equivalents thereof which may suggest themselves to those skilled in the art without departing from the spirit of the present disclosure and/or scope of the appended claims.

EXAMPLES

The application is further illustrated by the following examples, which should not be construed as further limiting. The practice of the present disclosure will employ, unless otherwise indicated, conventional techniques of organic synthesis, cell biology, cell culture, and molecular biology, which are within the skill of the art.

Abbreviations

ACN acetonitrile AcOH acetic acid PyBOP benzotriazol-1-yloxytripyrrolidinophosphonium h exafl uoro phosph ate

Cbz benzyl chlorocarbonate HDMS bis(trimethylsilyl)amide DEAD diethyl azodicarboxylate DIAD diisopropyl azodicarboxylate DCE dichloroethane DCM dichloromethane DIEA/DIPEA diisopropylethylamine DMAP 4-dimethylaminopyridine DMF dimethylformamide DTBAD di-tert-butyl azodicarboxylate EtOH ethanol EtOAc ethyl acetate EDCI 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide eq equivalent(s) g gram(s)

HPMA N-(2-Hydroxypropyl) methacrylamide 4hr hour(s) IPA isopropyl alcohol Lindlar catalyst H₂/Pd/CaCO₃ LC-MS liquid chromatography-mass spectrometry LAH lithium aluminum hydride m-CPBA meta-chloroperoxybenzoic acid Ms methanesulfonyl min minute(s) MS mass spectrometry MeOH methanol μL microliter(s) μm micrometer(s) μmol micromole(s) mg milligram(s) mm millimeter(s) mL milliliter(s) mmol millimole(s) NPht isoindoline-1 , 3-dione psi pounds per square inch

Py pyridine

TBDPS tert-butyldiphenylsilyl

Boc tert-butyloxycarbonyl

THF tetrahydrofuran

THP tetrahydropyran

TBAF tetra-n-butylammonium fluoride

Tol. toluene

PTSA p-toluenesulfonic acid

SFC supercritical fluid chromatography

Tos or Ts toluenesulfonyl

TEA triethylamine

TFA trifluoroacetic acid

Grubbs II (1 ,3-Bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene)dichloro

(phenylmethylene)(tricyclohexylphosphine)ruthenium

TLC thin layer chromatography Example 1: Synthetic Procedures

Procedure for Preparation of Compound 021

Scheme 1

To a solution of 4-amino-2-methylsulfanyl-pyrimidine-5-carbaldehyde (6.3 g, 37.5 mmol, 1 eq) and tert-butyl 4-(2-ethoxy-2-oxo-ethyl)-8-methyl-2,3-dihydroquinoxaline-1- carboxylate (12.5 g, 37.5 mmol, 1 eq) in DMF (150 mL) was added Cs₂CO₃ (36.6 g, 112.5 mmol, 3 eq). The mixture was stirred at 60°C for 2 hr. LC-MS showed desired compound was detected.

The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with H₂O (100 mL) and extracted with ethyl acetate (100 mL x 4). The combined organic layers were washed with brine (100 mL), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (80 g Silica Flash Column, Eluent of 0-20% Ethyl acetate/Petroleum ether gradient @ 100 mL/min) to give tert-butyl 8-methyl-4-(2- methylsulfanyl-7-oxo-8H-pyrido[2,3-d]pyrimidin-6-yl)-2,3-dihydroquinoxaline-1 -carboxylate (11 g, 25.0 mmol, 66.6% yield) as a yellow solid.

To a solution of tert-butyl 8-methyl-4-(2-methylsulfanyl-7-oxo-8H-pyrido[2,3- d]pyrimidin-6-yl)-2,3-dihydroquinoxaline-1-carboxylate (2 g, 4.6 mmol, 1 eq) in acetone (20 mL) was added 4-bromobutan-1-ol (2.1 g, 13.6 mmol, 3 eq), Nal (136.4 mg, 910.1 μmol, 0.2 eq) and Cs₂CO₃ (4.4 g, 13.6 mmol, 3 eq). The mixture was stirred at 60°C for 12 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (12 g Silica Flash Column, Eluent of 0-100% Ethyl acetate/Petroleum ether gradient @ 80 mL/min) to give tert-butyl 4-[8-(4-hydroxybutyl)-2-methylsulfanyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8- methyl-2,3-dihydroquinoxaline-1 -carboxylate (1.1 g, 2.1 mmol, 46.8% yield) as a yellow solid.

To a solution of tert-butyl 4-[8-(4-hydroxybutyl)-2-methylsulfanyl-7-oxo-pyrido[2,3- d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (1 .1 g, 2.1 mmol, 1 eq) in DCM (10 mL) was added MCPBA (519.0 mg, 2.6 mmol, 85% purity, 1 .2 eq) at 0°C. The mixture was stirred at 25°C for 12 hr. LC-MS showed desired compound was detected. The reaction mixture was quenched by addition Na₂SO₃ (20 mL) at 25°C, then diluted with dichloromethane (15 mL) and extracted with dichloromethane (20 mL x 3) and washed with NaHCO₃ (20 mL x 3), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give tert-butyl 4-[8-(4-hydroxybutyl)-2-methylsulfinyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8- methyl-2,3-dihydroquinoxaline-1 -carboxylate (1.2 g, crude) as a yellow solid.

To a solution of tert-butyl 4-[8-(4-hydroxybutyl)-2-methylsulfinyl-7-oxo-pyrido[2,3- d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (220 mg, 417.0 μmol, 1 eq) in dioxane (3 mL) was added 3-aminophenol (54.6 mg, 500.4 μmol, 1 .2 eq) TFA (61 .8 mg, 542.0 μmol, 40.3 μL, 1.3 eq). The mixture was stirred at 100°C for 12 hr. LC-MS showed desired compound was detected. The reaction was concentrated under reduce pressure. The residue was purified by flash silica gel chromatography (12 g Silica Flash Column, Eluent of 0-100% Ethyl acetate/Petroleum ether gradient @ 80 mL/min) to give tert-butyl 4- [2-(3-hydroxyanilino)-8-(4-hydroxybutyl)-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3- dihydroquinoxaline-1 -carboxylate (316 mg) as a yellow solid.

To a solution of tert-butyl 4-[2-(3-hydroxyanilino)-8-(4-hydroxybutyl)-7-oxo-pyrido[2,3- d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (316 mg, 551.8 μmol, 1 eq) in THF (15 mL) and Tol. (15 mL) was added DTBAD (508.3 mg, 2.2 mmol, 4 eq) PPh₃ (578.9 mg, 2.2 mmol, 4 eq) under N₂ atmosphere. The mixture was stirred at 70°C for 12 hr under N₂ atmosphere. LC-MS showed desired compound was detected. The reaction was concentrated under reduce pressure. The residue was purified by prep-TLC (SiO₂, petroleum ether : ethyl acetate=1 :1 ) to give tert-butyl 8-methyl-4-(14-oxo-8-oxa-2, 13, 19,20- tetrazatetracyclo[11.6.2.13,7.017,21]docosa-1(19),3,5,7(22),15,17,20-heptaen-15-yl)-2,3- dihydroquinoxaline-1-carboxylate (41 mg, 73.9 μmol, 13.4% yield) as a yellow solid.

A solution of tert-butyl 8-methyl-4-(14-oxo-8-oxa-2, 13, 19,20- tetrazatetracyclo[11.6.2.13,7.017,21]docosa-1(19),3,5,7(22),15,17,20-heptaen-15-yl)-2,3- dihydroquinoxaline-1-carboxylate (41 mg, 73.9 μmol, 1 eq) in HCI/EtOAc (1 mL, 4 M) was stirred at 25°C for 2 hr. LC-MS showed desired compound was detected. The reaction was concentrated under reduce pressure to give 15-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-8- oxa-2, 13,19,20-tetrazatetracyclo[11 .6.2.13,7.017,21 ]docosa- 1 ( 19), 3, 5, 7(22), 15,17,20- heptaen-14-one (38 mg, crude, HCI) as a yellow solid.

To a solution of 15-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-8-oxa-2,13,19,20- tetrazatetracyclo[11.6.2.13,7.017,21]docosa-1(19),3,5,7(22),15,17,20-heptaen-14-one (38 mg, 77.4 μmol, 1 eq, HCI) in DCM (2 mL) was added DIEA to adjust the pH to 7-8, then DIEA (30.0 mg, 232.2 μmol, 40.4 μL, 3 eq), prop-2-enoyl chloride (7.7 mg, 85.1 μmol, 6.9 μL, 1.1 eq) was added to the mixture at 0°C. The mixture was stirred at 0°C for 1 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (FA condition, column: Phenomenex Luna C 18 100 x 30 mm x 5 um; mobile phase: [H₂O(0.2% FA)- ACN]; gradient: 30 % - 70 % B over 8.0 min) to give 15-(5-methyl-4-prop-2-enoyl-2,3- dihydroquinoxalin-1 -yl)-8-oxa-2, 13,19,20- tetrazatetracyclo[11.6.2.13,7.017,21]docosa-1(19),3,5,7(22),15,17,20-heptaen-14-one (5.96 mg, 11.7 μmol, 15.1 % yield, 100% purity) as a white solid.

Procedure for Preparation of Compound 022 Scheme 2

To a solution of tert-butyl 8-methyl-4-(2-methylsulfanyl-7-oxo-8H-pyrido[2,3- d]pyrimidin-6-yl)-2,3-dihydroquinoxaline-1-carboxylate (1 g, 2.2 mmol, 1 eq) and 5- bromopentan-1-ol (570.0 mg, 3.4 mmol, 1.5 eq) in acetone (15 mL) was added Cs₂CO₃ (1.6 g, 5.0 mmol, 2.2 eq) and Nal (68.2 mg, 455.0 μmol, 0.2 eq). The mixture was stirred at 60°C for 12 hr. LC-MS showed the desired compound was detected. The reaction mixture was partitioned between H₂O (50 mL) and ethyl acetate (50 mL x 3). The organic phase was separated washed with brine (40 mL x 1), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (20 g Silica Flash Column, Eluent of 0-50% Ethyl acetate/Petroleum ether gradient @ 100 mL/min) to give tert-butyl 4-[8-(5-hydroxypentyl)-2-methylsulfanyl-7-oxo- pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (1 g, 1.9 mmol, 83.6% yield) as a yellow solid.

To a solution of tert-butyl 4-[8-(5-hydroxypentyl)-2-methylsulfanyl-7-oxo-pyrido[2,3- d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (950 mg, 1.8 mmol, 1 eq) in DCM (30 mL) was added m-CPBA (587.0 mg, 2.9 mmol, 85% purity, 1.6 eq) at 0°C. The mixture was stirred at 25°C for 2hr. LC-MS showed the desired compound was detected. The reaction mixture was partitioned between Na₂SO₃ (100 mL) and dichloromethane (100 mL x 3). The organic phase was separated washed with NaHCO₃ (50 mL x 3), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give tert-butyl 4-[8-(5- hydroxypentyl)-2-methylsulfonyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3- dihydroquinoxaline-1 -carboxylate (1 g, crude) as a yellow solid.

To a solution of tert-butyl 4-[8-(5-hydroxypentyl)-2-methylsulfonyl-7-oxo-pyrido[2,3- d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (180 mg, 322.7 μmol, 1 eq) in dioxane (2 mL) was added 3-aminophenol (52.8 mg, 484.1 μmol, 1 .5 eq) and TFA (55.2 mg, 484.1 μmol, 35.9 μL, 1 .5 eq). The mixture was stirred at 80°C for 12 hr. LC-MS showed the desired compound was detected. The reaction mixture was partitioned between NaHCO₃ (60 mL) and ethyl acetate (80 mL x 3). The organic phase was separated washed with brine (50 mL x 1), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (20 g Silica Flash Column, Eluent of 0-50% Ethyl acetate/Petroleum ether gradient @ 120 mL/min) to give tert-butyl 4-[2-(3-hydroxyanilino)-8-(5-hydroxypentyl)-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]- 8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (650 mg, 926.2 μmol, 57.4% yield, 83.6% purity) as a yellow solid.

To a solution of tert-butyl 4-[2-(3-hydroxyanilino)-8-(5-hydroxypentyl)-7-oxo- pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (300 mg, 511.3 μmol, 1 eq) and DTBAD (470.9 mg, 2.0 mmol, 4 eq) in THF (15 mL) and Tol. (15 mL) was added PPh₃ (536.4 mg, 2.0 mmol, 4 eq). The mixture was stirred at 25°C for 12 hr under N₂. LC-MS showed the desired compound was detected. The reaction mixture was partitioned between H₂O (50 mL) and ethyl acetate (50 mL x 3). The organic phase was separated washed with brine (30 mL x 1 ), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiO₂, Petroleum ether : Ethyl acetate = 1 :1 ) to give tert-butyl 8-methyl-4-(1⁷-oxo-1⁷,1⁸-dihydro-4-oxa-2-aza-1(2,8)- pyrido[2,3-d]pyrimidina-3(1 ,3)-benzenacyclononaphane-1⁶-yl)-3,4-dihydroquinoxaline-1 (2H)- carboxylate (70 mg, 123.1 μmol, 24.1 % yield) as a yellow solid.

To a solution of tert-butyl 8-methyl-4-(1⁷-oxo-1⁷,1⁸-dihydro-4-oxa-2-aza-1 (2,8)- pyrido[2,3-d]pyrimidina-3(1 ,3)-benzenacyclononaphane-1⁶-yl)-3,4-dihydroquinoxaline-1 (2H)- carboxylate (70 mg, 123.1 μmol, 1 eq) in EtOAc (1 mL) was added HCI/EtOAc (0.5 mL, 4M). The mixture was stirred at 25 °C for 2 hr. LC-MS showed the desired compound was detected. The reaction mixture was concentrated under reduced pressure to give 1⁶-(5- methyl-3,4-dihydroquinoxalin-1 (2H)-yl)-1⁷,1⁸-dihydro-4-oxa-2-aza-1(2,8)-pyrido[2,3- d]pyrimidina-3(1 ,3)-benzenacyclononaphan-1⁷-one (70 mg, crude) as a yellow solid.

To a solution of 1⁶-(5-methyl-3,4-dihydroquinoxalin-1 (2H)-yl)-1⁷,1⁸-dihydro-4-oxa-2- aza-1 (2,8)-pyrido[2,3-d]pyrimidina-3(1 ,3)-benzenacyclononaphan-1⁷-one (60 mg, 128.0 μmol, 1 eq) in DCM (2 mL) was added TEA (38.8 mg, 384.1 μmol, 53.4 μL, 3 eq) and prop- 2-enoyl chloride (11 .6 mg, 128.0 μmol, 10.4 μL, 1 eq) at 0°C. The mixture was stirred at 0°C for 1 hr. LC-MS showed the desired compound was detected. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep- HPLC (TFA condition, column: Phenomenex luna C₁₈100 x 40 mm x 5 um; mobile phase: [H₂O (0.1 % TFA) - ACN]; gradient : 50%-80% B over 8.0 min) to give 1⁶-(4-acryloyl-5- methyl-3,4-dihydroquinoxalin-1 (2H)-yl)-1⁷,1⁸-dihydro-4-oxa-2-aza-1(2,8)-pyrido[2,3- d]pyrimidina-3(1 ,3)-benzenacyclononaphan-1⁷-one (35.4 mg , 98.2% purity) as a white solid.

Procedure of Compound 023

Scheme 3

To a solution of tert-butyl 8-methyl-4-(2-methylsulfanyl-7-oxo-8H-pyrido[2,3- d]pyrimidin-6-yl)-2,3-dihydroquinoxaline-1-carboxylate (2.5 g, 5.6 mmol, 1 eq) and 6- bromohexan-1-ol (1.2 g, 6.8 mmol, 892.9 μL, 1.2 eq) in acetone (80 mL) was added Cs₂CO₃ (5.5 g, 17.0 mmol, 3 eq) and Nal (85.2 mg, 568.7 μmol, 0.1 eq). The mixture was stirred at 60°C for 4 hr. LC-MS showed desired compound was detected. The reaction mixture was filtered and concentrated under reduced pressure to remove solvent. The residue was purified by flash silica gel chromatography (12 g Silica Flash Column, Eluent of 0-19% Ethyl acetate/Petroleum ether gradient @ 100 mL/min) to give tert-butyl 4-[8-(6-hydroxyhexyl)-2- methylsulfanyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1- carboxylate (1.6 g, 2.8 mmol, 49.5% yield, 95% purity) as a yellow solid.

At 0°C, a solution of tert-butyl 4-[8-(6-hydroxyhexyl)-2-methylsulfanyl-7-oxo- pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (1.6 g, 2.9 mmol, 1 eq) in DCM (15 mL) was added m-CPBA (782.4 mg, 3.8 mmol, 85% purity, 1.3 eq). The mixture was stirred at 20°C for 2 hr. LC-MS showed desired compound was detected. The reaction mixture was quenched by addition Na₂SO₃ (30 mL), and then extracted with dichloromethane (20 mL x 3). The combined organic layers were washed with NaHCO₃ (20 mL x 3), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give tertbutyl 4-[8-(6-hydroxyhexyl)-2-methylsulfonyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3- dihydroquinoxaline-1 -carboxylate (1 .6 g, crude) as an orange solid.

To a solution of tert-butyl 4-[8-(6-hydroxyhexyl)-2-methylsulfonyl-7-oxo-pyrido[2,3- d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (800 mg, 1.4 mmol, 1 eq) in dioxane (8 mL) was added 3-aminophenol (229.0 mg, 2.1 mmol, 1.5 eq) and TFA (239.3 mg, 2.1 mmol, 155.9 μL, 1.5 eq). The mixture was stirred at 100°C for 2 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by flash silica gel chromatography (20 g Silica Flash Column, Eluent of 0-47% Ethyl acetate/Petroleum ether gradient @ 100 mL/min) to give tert-butyl 4-[2-(3-hydroxyanilino)-8-(6-hydroxyhexyl)-7-oxo-pyrido[2,3- d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (300 mg, 424.5 μmol, 30.3% yield, 85% purity) as a yellow solid.

A mixture of tert-butyl 4-[2-(3-hydroxyanilino)-8-(6-hydroxyhexyl)-7-oxo-pyrido[2,3- d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (170 mg, 283.0 μmol, 1 eq), PPh₃ (296.9 mg, 1.1 mmol, 4 eq), DTBAD (260.6 mg, 1.1 mmol, 4 eq) in THF (8.5 mL) and Tol. (8.5 mL) was degassed and purged with N₂ for 3 times, and then the mixture was stirred at 20°C for 12 hr under N₂ atmosphere. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-TLC (SiO₂, Petroleum ether/Ethyl acetate=1/1 ) to give tert-butyl 8-methyl-4-(16-oxo-8-oxa-2, 15,21 ,22-tetrazatetracyclo[13.6.2.1^3,7.0^19,23]tetracosa- 1 (21 ),3,5,7(24),17,19,22-heptaen-17-yl)-2,3-dihydroquinoxaline-1-carboxylate (50 mg, 72.0 μmol, 25.4% yield, 84% purity) as a yellow solid.

A solution of tert-butyl 8-methyl-4-(16-oxo-8-oxa-2, 15,21 ,22- tetrazatetracyclo[13.6.2.1^3,7.0^19,23]tetracosa-1 (21 ),3,5,7(24),17,19,22-heptaen-17-yl)-2,3- dihydroquinoxaline-1 -carboxylate (50 mg, 85.8 μmol, 1 eq) in HCI/EtOAc (0.7 mL, 4 M) and EtOAc (0.5 mL) was stirred at 20°C for 1 hr. LC-MS showed desired compound was detected.

The reaction mixture was concentrated under reduced pressure to give 17-(5-methyl-3,4- dihydro-2H-quinoxalin-1-yl)-8-oxa-2, 15,21 ,22-tetrazatetracyclo[13.6.2.1^3,7.0^19,23]tetracosa- 1 (21 ),3,5,7(24),17,19,22-heptaen-16-one (49 mg, crude, HCI) as a yellow solid.

To a solution of 17-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-8-oxa-2,15,21 ,22- tetrazatetracyclo[13.6.2.1^3,7.0^19,23]tetracosa-1 (21 ), 3, 5, 7(24), 17, 19,22-heptaen-16-one (49 mg, 94.4 μmol, 1 eq, HCI) in DCM (2 mL) was added dropwise TEA (28.6 mg, 283.2 μmol, 39.4 μL, 3 eq) and then prop-2-enoyl chloride (8.5 mg, 94.4 μmol, 7.6 μL, 1 eq) in DCM (1 mL) was added dropwise at 0°C. The mixture was stirred at 0°C for 30 min. LC-MS showed desired compound was detected. The reaction mixture was concentrated under N₂ to remove solvent. The residue was purified by prep-HPLC (TFA condition, column: Phenomenex luna C18 100x40mmx5 um; mobile phase: [H₂O(0.1% TFA)- ACN];gradient:55%-85% B over 8.0 min) to give 17-(5-methyl-4-prop-2-enoyl-2,3- dihydroquinoxalin-1 -yl)-8-oxa-2, 15,21 ,22-tetrazatetracyclo[13.6.2.13,7.019, 23]tetracosa- 1 (21 ),3,5,7(24),17,19,22-heptaen-16-one (19.8 mg, 36.9 μmol, 99.2% yield, 100% purity) as a yellow solid. Procedure for Preparation of Compound 007

Scheme 4

To a solution of but-3-yn-1-ol (5 g, 71.3 mmol, 5.4 mL, 1 eq) in DCM (100 mL) was added DMAP (871 .5 mg, 7.1 mmol, 0.1 eq) and TEA (7.2 g, 71.3 mmol, 9.9 mL, 1 eq). Then the mixture was added tert-butyl-chloro-diphenyl-silane (19.6 g, 71 .3 mmol, 18.3 mL, 1 eq) at 0°C under N₂ atmosphere. The mixture was stirred at 25°C for 12 hr under N₂ atmosphere. TLC indicated one new spot formed. The residue was diluted with H₂O (70 mL) and extracted with ethyl acetate (100 mL x 4). The combined organic layers were washed with brine (80 mL x 1 ), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (40 g Silica Flash Column, Eluent of 0-0% Ethyl acetate/Petroleum ether gradient @ 120 mL/min) to give tert-butyl-but-3-ynoxy-diphenyl-silane (21 .2 g, 68.7 mmol, 96.2% yield) as a white oil.

To a solution of tert-butyl-but-3-ynoxy-diphenyl-silane (14 g, 45.4 mmol, 1 eq) and HMPA (8.1 g, 45.4 mmol, 7.9 mL, 1 eq) in THF (210 mL) was added n-BuLi (1.6 M, 56.7 mL, 2 eq) at -78°C for 30 min under N₂ atmosphere. Then the mixture was added oxirane (20.2 g, 453.8 mmol, 22.9 mL, 10 eq) at -78°C under N₂ atmosphere. The mixture was stirred at 25°C for 12 hr under N₂ atmosphere. TLC indicated one new spot formed. The reaction mixture was quenched by addition NH₄CI (150 mL) at 0 °C, and extracted with ethyl acetate (200 mL x 4). The combined organic layers were washed with brine (180 mL x 2), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (220 g Silica Flash Column, Eluent of 0-16% Ethyl acetate/Petroleum ether gradient @ 70 mL/min) to give 6-[tert- butyl(diphenyl)silyl]oxyhex-3-yn-1-ol (9.3 g, 26.4 mmol, 58.1 % yield) as a light yellow oil.

To a solution of Lindlar catalyst (5 g, 10% purity) in MeOH (80 mL) was added 6-[tert- butyl(diphenyl)silyl]oxyhex-3-yn-1-ol (9.9 g, 27.9 mmol, 1 eq) under N₂ atmosphere. The suspension was degassed and purged with H₂ for 3 times. The mixture was stirred under H₂ (15 Psi) at 25 °C for 4 hr. Then the reaction mixture filtered and concentrated under reduced pressure to give a residue. To a solution of Lindlar catalyst (5 g, 10% purity) in MeOH (80 mL) was added 6-[tert-butyl(diphenyl)silyl]oxyhex-3-yn-1-ol (9.9 g, 27.9 mmol, 1 eq) under N₂ atmosphere. The suspension was degassed and purged with H₂ for 3 times. The mixture was stirred under H₂ (15 Psi) at 25 °C for 1 hr. LC-MS showed desired compound was detected. The reaction mixture filtered and concentrated under reduced pressure to give (Z)-6-[tert- butyl(diphenyl)silyl]oxyhex-3-en-1-ol (9.9 g, crude) as a light yellow oil.

To a solution of (Z)-6-[tert-butyl(diphenyl)silyl]oxyhex-3-en-1-ol (9.9 g, 27.9 mmol, 1 eq) in DCM (300 mL) was added CBr₄ (27.8 g, 83.8 mmol, 3 eq) under N₂ atmosphere. Then the mixture was added PPh₃ (22.0 g, 83.8 mmol, 3 eq) in DCM (30 mL) at 0°C under N₂ atmosphere. The mixture was stirred at 25°C for 12 hr. TLC indicated one new spot formed. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by flash silica gel chromatography (120 g Silica Flash Column, Eluent of 0~3% Ethyl acetate/Petroleum ether gradient @ 70 mL/min) to give [(Z)-6- bromohex-3-enoxy]-tert-butyl-diphenyl-silane (10.4 g, 24.9 mmol, 89.2% yield) as a yellow oil.

To a solution of tert-butyl 8-methyl-4-(2-methylsulfanyl-7-oxo-8H-pyrido[2,3- d]pyrimidin-6-yl)-2,3-dihydroquinoxaline-1-carboxylate (4 g, 9.1 mmol, 1 eq) in DCM (40 mL) was added m-CPBA (2.2 g, 10.9 mmol, 85% purity, 1 .2 eq) at 0°C. The mixture was stirred at 25°C for 1 hr. LC-MS showed desired compound was detected. The residue was diluted with Na₂SO₃ (70 mL) and extracted with dichloromethane (100 mL x 3). The combined organic layers were washed with NaHCO₃ (70 mL x 3), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give tert-butyl 8-methyl-4-(2-methylsulfinyl-7-oxo- 8H-pyrido[2,3-d]pyrimidin-6-yl)-2,3-dihydroquinoxaline-1 -carboxylate (4 g, crude) as a yellow solid.

To a solution of tert-butyl 8-methyl-4-(2-methylsulfinyl-7-oxo-8H-pyrido[2,3- d]pyrimidin-6-yl)-2,3-dihydroquinoxaline-1-carboxylate (4 g, 8.8 mmol, 1 eq) in IPA (50 mL) was added 1-tetrahydropyran-2-ylpyrazol-4-amine (2.9 g, 17.6 mmol, 2 eq). The mixture was stirred at 80°C for 12 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The reaction was triturated with methyl tertiary ether at 25°C for 1.5 hr. The reaction mixture was filtered and the filter cake dried in vacuum to give tert-butyl 8-methyl-4-[7-oxo-2-[(1-tetrahydropyran- 2-ylpyrazol-4-yl)amino]-8H-pyrido[2,3-d]pyrimidin-6-yl]-2,3-dihydroquinoxaline-1 -carboxylate (5.0 g, 7.8 mmol, 89.1% yield, 87.9% purity) as a yellow solid.

To a solution of tert-butyl 8-methyl-4-[7-oxo-2-[(1-tetrahydropyran-2-ylpyrazol-4- yl)amino]-8H-pyrido[2,3-d]pyrimidin-6-yl]-2,3-dihydroquinoxaline-1 -carboxylate (3 g, 5.4 mmol, 1 eq) in Acetone (30 mL) was added [(Z)-6-bromohex-3-enoxy]-tert-butyl-diphenyl- silane (2.7 g, 6.4 mmol, 1.2 eq), Cs₂CO₃ (5.3 g, 16.1 mmol, 3 eq) and Nal (161.0 mg, 1.1 mmol, 0.2 eq). The mixture was stirred at 60°C for 12 hr. LC-MS showed desired compound was detected.

The residue was diluted with H₂O (50 mL) and extracted with ethyl acetate (80 mL x 3). The combined organic layers were washed with brine (50 mL x 1), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (40 g Silica Flash Column, Eluent of 0-41% Ethyl acetate/Petroleum ether gradient @ 100 mL/min) to give tert-butyl 4-[8-[(Z)-6-[tert- butyl(diphenyl)silyl]oxyhex-3-enyl]-7-oxo-2-[(1-tetrahydropyran-2-ylpyrazol-4- yl)amino]pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (990 mg, 1.1 mmol, 20.6% yield) as a yellow solid.

To a solution of tert-butyl 4-[8-[(Z)-6-[tert-butyl(diphenyl)silyl]oxyhex-3-enyl]-7-oxo-2- [(1-tetrahydropyran-2-ylpyrazol-4-yl)amino]pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3- dihydroquinoxaline-1-carboxylate (900 mg, 1.0 mmol, 1 eq) in THF (9 mL) was added TBAF (1 M, 2.01 mL, 2 eq). The mixture was stirred at 25°C for 1 hr. LC-MS showed desired compound was detected. The residue was diluted with H₂O (20 mL) and extracted with ethyl acetate (30 mL x 4). The combined organic layers were washed with brine (20 mL x 1 ), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (4 g Silica Flash Column, Eluent of 0-55% Ethyl acetate/Petroleum ether gradient @ 100 mL/min) to give tert-butyl 4-[8-[(Z)-6- hydroxyhex-3-enyl]-7-oxo-2-[(1-tetrahydropyran-2-ylpyrazol-4-yl)amino]pyrido[2,3- d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (430 mg, 654.7 μmol, 65.1 % yield) as a yellow solid.

To a solution of tert-butyl 4-[8-[(Z)-6-hydroxyhex-3-enyl]-7-oxo-2-[(1-tetrahydropyran- 2-ylpyrazol-4-yl)amino]pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1- carboxylate (430 mg, 654.7 μmol, 1 eq) in DCM (5 mL) was added TEA (331.3 mg, 3.3 mmol, 455.6 μL, 5 eq), DMAP (8.0 mg, 65.4 μmol, 0.1 eq) and TosCI (374.5 mg, 2.0 mmol, 3 eq). The mixture was stirred at 25°C for 1 hr. LC-MS showed desired compound was detected. The residue was diluted with NaHCO₃ (15 mL) and extracted with dichloromethane (20 mL x 4), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (12 g Silica Flash Column, Eluent of 0-48% Ethyl acetate/Petroleum ether gradient @ 100 mL/min) to give tert-butyl 8-methyl-4-[7-oxo-8-[(Z)-6-(p-tolylsulfonyloxy)hex-3-enyl]-2-[(1- tetrahydropyran-2-ylpyrazol-4-yl)amino]pyrido[2,3-d]pyrimidin-6-yl]-2,3-dihydroquinoxaline-1- carboxylate (350 mg, 431.6 μmol, 65.9% yield) as a yellow solid.

To a solution of tert-butyl 8-methyl-4-[7-oxo-8-[(Z)-6-(p-tolylsulfonyloxy)hex-3-enyl]-2- [(1-tetrahydropyran-2-ylpyrazol-4-yl)amino]pyrido[2,3-d]pyrimidin-6-yl]-2,3- dihydroquinoxaline-1-carboxylate (300 mg, 369.9 μmol, 1 eq) in MeOH (3 mL) was added PTSA (127.4 mg, 739.9 μmol, 2 eq). The mixture was stirred at 25°C for 1 hr. LC-MS showed desired compound was detected. The residue was diluted with NaHCO₃ (10 mL) and extracted with ethyl acetate (15 mL x 3). The combined organic layers were washed with brine (10 mL x 2), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (4 g Silica Flash Column, Eluent of 0-50% Ethyl acetate/Petroleum ether gradient @ 100 mL/min) to give tert-butyl 8-methyl-4-[7-oxo-8-[(Z)-6-(p-tolylsulfonyloxy)hex-3-enyl]-2-(1 H-pyrazol-4- ylamino)pyrido[2,3-d]pyrimidin-6-yl]-2,3-dihydroquinoxaline-1 -carboxylate (238 mg, 327.4 μmol, 88.5% yield) as a yellow solid.

To a solution of tert-butyl 8-methyl-4-[7-oxo-8-[(Z)-6-(p-tolylsulfonyloxy)hex-3-enyl]-2- (1 H-pyrazol-4-ylamino)pyrido[2,3-d]pyrimidin-6-yl]-2,3-dihydroquinoxaline-1 -carboxylate (238 mg, 327.4 μmol, 1 eq) in DMF (3 mL) was added K₂CO₃ (135.8 mg, 982.3 μmol, 3 eq). The mixture was stirred at 80°C for 12 hr. LC-MS showed desired compound was detected. The residue was diluted with H₂O (10 mL) and extracted with ethyl acetate (20 mL x 4). The combined organic layers were washed with brine (15 mL x 2), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiO₂, Petroleum ether : Ethyl acetate = 0:1 ) to give tert-butyl 8-methyl-4-[(9Z)-14- oxo-2, 5, 6, 13, 19, 20-hexazatetracyclo[11.6.2.13, 6.017, 21]docosa-1 (19), 3(22), 4, 9, 15, 17,20- heptaen-15-yl]-2,3-dihydroquinoxaline-1 -carboxylate (100 mg, 180.3 μmol, 55.1% yield) as a yellow solid.

A mixture of tert-butyl 8-methyl-4-[(9Z)-14-oxo-2,5,6,13,19,20- hexazatetracyclo[11 ,6.2.13,6.017,21]docosa-1 (19),3(22),4,9,15,17,20-heptaen-15-yl]-2,3- dihydroquinoxaline-1 -carboxylate (80 mg, 144.2 μmol, 1 eq) in EtOAc (1 mL) and HCI/EtOAc (1 mL, 4M) was stirred at 25°C for 0.5 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent to give (9Z)-15-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-2,5,6, 13, 19,20- hexazatetracyclo[11 ,6.2.13,6.017,21]docosa-1 (19),3(22),4,9,15,17,20-heptaen-14-one (80 mg, crude, HCI) as a yellow solid.

To a solution of (9Z)-15-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-2,5,6,13,19,20- hexazatetracyclo[11 ,6.2.13,6.017,21]docosa-1 (19),3(22),4,9,15,17,20-heptaen-14-one (80 mg, 162.9 μmol, 1 eq, HCI) in DCM (2 mL) was added TEA (49.5 mg, 488.8 μmol, 68.0 μL, 3 eq) and prop-2-enoyl chloride (14.8 mg, 162.9 μmol, 13.2 μL, 1 eq) at 0°C. The mixture was stirred at 0°C for 0.5hr. LC-MS showed 20% Reactant 1 was remained. Then the mixture was added prop-2-enoyl chloride (4.4 mg, 48.9 μmol, 4.0 μL, 0.3 eq) at 0°C. The mixture was stirred at 0°C for 0.5 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (TFA condition, column: Phenomenex Luna C18 100 x 30 mm x 5 um; mobile phase: [H₂O(0.1% TFA)-ACN]; gradient:30%-60% B over 8.0 min) to give (9Z)- 15-(5-methyl-4-prop-2-enoyl-2,3-dihydroquinoxalin-1 -yl)-2,5,6, 13,19,20- hexazatetracyclo[11 ,6.2.13,6.017,21]docosa-1 (19),3(22),4,9,15,17,20-heptaen-14-one (20.77 mg, 39.43 μmol, 24.20% yield, 96.56% purity) as a yellow solid. VT ¹H NMR (400 MHz, DMSO-d₆) δ = 9.36 - 9.22 (m, 1 H), 8.67-8.62 (m, 1 H), 8.16 (s, 1 H), 7.83 (s, 1 H), 7.25 (s, 1 H), 6.88 (br t, J = 7.9 Hz, 1 H), 6.63 (br d, J = 7.3 Hz, 1 H), 6.37 - 6.20 (m, 3H), 5.75 - 5.66 (m, 1H), 5.60 - 5.52 (m, 1H), 5.44 - 5.32 (m, 1H), 5.02 - 4.74 (m, 1H), 4.20 (td, J = 2.6, 5.2 Hz, 2H), 4.13 - 3.90 (m, 3H), 3.77 - 3.68 (m, 1H), 3.14 - 2.97 (m, 1H), 2.58 (brt, J= 8.6 Hz, 2H), 2.46 (brd, J= 9.5 Hz, 2H), 2.13 (s, 3H).

Procedure for Preparation of Compound 018

Scheme 5

To a solution of tert-butyl 8-methyl-4-(2-methylsulfanyl-7-oxo-8H-pyrido[2,3- d]pyrimidin-6-yl)-2,3-dihydroquinoxaline-1-carboxylate (655 mg, 1.4 mmol, 1eq) in DCM (10 mL) was added m-CPBA (453.8 mg, 2.2 mmol, 85% purity, 1.5 eq) at 0°C. The resulting mixture was stirred at 25°C for 2 hr. LC-MS showed desired compound was detected. The reaction mixture was quenched by addition Na₂SO₃ (20 mL), extracted with dichloromethane (20 mL x 3). The combined organic layers were washed with NaHCO₃ (20 mL x 2), and dried over Na₂SO₄, filtered and concentrated under reduced pressure to give tert-butyl 8- methyl-4-(2-methylsulfinyl-7-oxo-8H-pyrido[2,3-d]pyrimidin-6-yl)-2,3-dihydroquinoxaline-1- carboxylate (715 mg, crude) as a red solid.

To a solution of tert-butyl 8-methyl-4-(2-methylsulfinyl-7-oxo-8H-pyrido[2,3- d]pyrimidin-6-yl)-2,3-dihydroquinoxaline-1-carboxylate (358.4 mg, 786.7 μmol, 1 eq) in dioxane (5 mL) was added TFA (134.5 mg, 1.1 mmol, 87.6 μL, 1 .5 eq) and 1-but-3- enylpyrazol-4-amine (161.9 mg, 1.1 mmol, 1.5 eq). The mixture was stirred at 100°C for 2 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The crude product was triturated with ethyl acetate (10 mL) at 25°C for 1 hr to give tert-butyl 4-[2-[(1-but-3-enylpyrazol-4- yl)amino]-7-oxo-8H-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1- carboxylate (522.4 mg, 958.6 μmol, 60.9% yield, 97% purity) as yellow oil.

To a solution of tert-butyl 4-[2-[(1-but-3-enylpyrazol-4-yl)amino]-7-oxo-8H-pyrido[2,3- d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (522.4 mg, 988.2 μmol, 1 eq) in DMF (7 mL) was added 4-bromobut-1-ene (133.4 mg, 988.2 μmol, 100.3 μL, 1 eq) and NaH (47.4 mg, 1.1 mmol, 7.5e^-1 μL, 60% purity, 1 .2 eq) at 0°C. The mixture was stirred at 25°C for 12 hr. LC-MS showed desired compound was detected. The residue was diluted with NH₄CI (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with brine (20 mL x 2), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (20 g Silica Flash Column, Eluent of 0-100% Ethyl acetate/Petroleum ether gradient @ 100 mL/min) to give tert-butyl 4-[8-but-3-enyl-2-[(1-but- 3-enylpyrazol-4-yl)amino]-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3- dihydroquinoxaline-1-carboxylate (342.6 mg, 517.4 μmol, 52.3% yield, 88% purity) as a yellow solid.

To a solution of tert-butyl 4-[8-but-3-enyl-2-[(1-but-3-enylpyrazol-4-yl)amino]-7-oxo- pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (239.4 mg, 410.8 μmol, 1 eq) in Tol. (100 mL) was degassed and purged with N₂ for 3 times, and then the mixture was stirred at 110°C for 1 hr under N₂ atmosphere. Benzylidene-[1 ,3-bis(2,4,6- trimethylphenyl)imidazolidin-2-ylidene]-dichlororuthenium;tricyclohexylphosphane (104.6 mg, 123.2 μmol, 0.3 eq) in Tol. (20 mL) was added and then the mixture was stirred at 110°C for 12 hr under N₂ atmosphere. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (TFA condition; column: Phenomenex luna C18 100x40mmx5 um; mobile phase: [H₂O(0.1% TFA)-ACN];gradient:35%-65% B over 8.0 min) to give tert-butyl 8- methyl-4-[(9Z)-14-oxo-2,5,6, 13,19,20-hexazatetracyclo[11 .6.2.13,6.017,21 ]docosa- 1 (19), 3(22), 4, 9, 15,17, 20-heptaen-15-yl]-2,3-dihydroquinoxaline-1 -carboxylate (94 mg, 169.4 μmol, 41 .2% yield) as a yellow solid.

A solution of tert-butyl 8-methyl-4-[(9Z)-14-oxo-2,5,6,13,19,20- hexazatetracyclo[11 ,6.2.13,6.017,21]docosa-1 (19),3(22),4,9,15,17,20-heptaen-15-yl]-2,3- dihydroquinoxaline-1 -carboxylate (94 mg, 169.4 μmol, 1 eq) in HCI/EtOAc (1 mL, 4M) and EtOAc (1 mL) was stirred at 25°C for 1 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to give (9Z)-15-(5-methyl- 3,4-dihydro-2H-quinoxalin-1-yl)-2,5,6,13,19,20-hexazatetracyclo[11 .6.2.13,6.017,21 ]docosa- 1 (19),3(22),4,9,15,17,20-heptaen-14-one (100 mg, crude, HCI) as a yellow solid.

To a solution of (9Z)-15-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-2,5,6,13,19,20- hexazatetracyclo[11 ,6.2.13,6.017,21]docosa-1 (19),3(22),4,9,15,17,20-heptaen-14-one (50 mg, 101 .8 μmol, 1 eq, HCI) in DCM (2 mL) was added TEA (30.9 mg, 305.5 μmol, 42.5 μL, 3 eq). And then prop-2-enoyl chloride (4.6 mg, 50.9 μmol, 4.1 μL, 0.5 eq) in DCM (0.1 mL) was added dropwise at 0°C. The mixture was stirred at 0°C for 0.5 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (TFA condition; column: Phenomenex luna C18 100x40mmx5 um; mobilephase: [H₂O(0.1% TFA)- ACN];gradient:25%-55% B over 8.0 min) to give (9Z)-15-(5-methyl-4-prop-2-enoyl-2,3- dihydroquinoxalin-1 -yl)-2,5,6, 13, 19,20-hexazatetracyclo[11 .6.2.13,6.017,21]docosa- 1 (19),3(22),4,9,15,17,20-heptaen-14-one (6.3 mg, 10.1 μmol, 9.9% yield, 100% purity, TFA) as a yellow solid.

Procedure for Preparation of Compound 024

Scheme 6

To a solution of tert-butyl 8-methyl-4-(2-methylsulfanyl-7-oxo-8H-pyrido[2,3- d]pyrimidin-6-yl)-2,3-dihydroquinoxaline-1-carboxylate (2.3 g, 5.2 mmol, 1 eq) in acetone (35 mL) was added Nal (235.3 mg, 1 .6 mmol, 0.3 eq) Cs₂CO₃ (5.1 g, 15.7 mmol, 3 eq) and 2-(2- bromoethoxy)ethanol (1.3 g, 7.9 mmol, 1.5 eq). The mixture was stirred at 60°C for 12 hr. LC-MS showed desired compound was detected. The reaction mixture was partitioned between H₂O (400 mL) and ethyl acetate (400 mL x 3). The organic phase was separated, dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (12 g Silica Flash Column, Eluent of 0-40% Ethyl acetate/Petroleum ether gradient @ 100 mL/min) to give tert-butyl 4-[8-[2-(2- hydroxyethoxy)ethyl]-2-methylsulfanyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3- dihydroquinoxaline-1-carboxylate (1.7 g, 3.2 mmol, 61.6% yield) as a yellow solid.

To a solution of tert-butyl 4-[8-[2-(2-hydroxyethoxy)ethyl]-2-methylsulfanyl-7-oxo- pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (1.7 g, 3.2 mmol, 1 eq) in DCM (17 mL) was added m-CPBA (654.1 mg, 3.2 mmol, 85% purity, 1 eq) at 0°C. The mixture was stirred at 0°C for 2 hr. LC-MS showed desired compound was detected. The reaction mixture was partitioned between Na₂SO₃ (400 mL) and dichloromethane (400 x 3 mL). The organic phase was separated, washed with NaHCO₃ (500 mL x 3), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give tert-butyl 4-[8-[2-(2- hydroxyethoxy)ethyl]-2-methylsulfinyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3- dihydroquinoxaline-1 -carboxylate (1 .5 g, crude) was obtained as a yellow solid.

To a solution of tert-butyl 4-[8-[2-(2-hydroxyethoxy)ethyl]-2-methylsulfinyl-7-oxo- pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (300 mg, 551.8 μmol, 1 eq) in dioxane (3 mL) was added TFA (94.3 mg, 827.7 μmol, 61.5 μL, 1 .5 eq) and 3- aminophenol (60.2 mg, 551 .8 μmol, 1 eq). The mixture was stirred at 80°C for 12 hr. LC- MS showed desired compound was detected. The reaction mixture was partitioned between NaHCO₃ (400 mL) and ethyl acetate (400 mL x 3). The organic phase was separated, washed with brine (400 mL x 2), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (12 g Silica Flash Column, Eluent of 0-100% Ethyl acetate/Petroleum ether gradient @ 100 mL/min) to give tert-butyl 4-[2-(3-hydroxyanilino)-8-[2-(2-hydroxyethoxy)ethyl]-7-oxo- pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (770 mg, 1.3 mmol, 47.4% yield) as a yellow solid.

To a solution of tert-butyl 4-[2-(3-hydroxyanilino)-8-[2-(2-hydroxyethoxy)ethyl]-7-oxo- pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (720 mg, 1.2 mmol, 1 eq) in THF (80 mL) was added PPh₃ (962.4 mg, 3.7 mmol, 3 eq), DEAD (639.0 mg, 3.7 mmol, 667.1 μL, 3 eq) and Na₂SO₄ (173.7 mg, 1 .2 mmol, 124.1 μL, 1 eq). The mixture was stirred at 50°C for 2 hr under N₂ atmosphere. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-TLC (SiO₂, Petroleum ether/Ethyl acetate=1/1 ) to give tert-butyl 8-methyl-4-(15-oxo-8,11-dioxa-2,14,20,21- tetrazatetracyclo[12.6.2.13,7.018,22]tricosa-1 (20),3,5,7(23),16,18,21-heptaen-16-yl)-2,3- dihydroquinoxaline- 1 -carboxylate (360 mg) as a yellow solid.

To a solution of tert-butyl 8-methyl-4-(15-oxo-8,11-dioxa-2, 14, 20,21- tetrazatetracyclo[12.6.2.13,7.018,22]tricosa-1 (20), 3, 5, 7(23), 16, 18,21 -heptaen-16-yl)-2,3- dihydroquinoxaline-1-carboxylate (200 mg, 350.5 μmol, 1 eq) in EtOAc (1.5 mL) was added HCI/EtOAc (1.5 mL, 4M) at 0°C. The mixture was stirred at 0°C for 1.5 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to give 16-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-8,11-dioxa-2,14,20,21- tetrazatetracyclo[12.6.2.13,7.018,22]tricosa-1(20),3,5,7(23),16,18,21-heptaen-15-one (200 mg, crude) as a yellow solid.

To a solution of 16-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-8,11-dioxa-2,14,20,21- tetrazatetracyclo[12.6.2.13,7.018,22]tricosa-1(20),3,5,7(23),16,18,21-heptaen-15-one (350 mg, 743.9 μmol, 1 eq) in DCM (7 mL) was added TEA (376.4 mg, 3.72 mmol, 517.7 μL, 5 eq) and prop-2-enoyl chloride (67.3 mg, 743.9 μmol, 60.4 μL, 1 eq) at 0°C. The mixture was stirred at 0°C for 15min. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (TFA condition column: Phenomenex Luna C1875 x 30mm x 3um; mobile phase: [H₂O(0.1% TFA)-ACN]; gradient: 40% - 70% B over 8.0 min) to give 16-(5- methyl-4-prop-2-enoyl-2,3-dihydroquinoxalin-1 -yI)-8 , 11 -dioxa-2, 14,20,21- tetrazatetracyclo[12.6.2.13,7.018,22]tricosa-1(20),3,5,7(23),16,18,21-heptaen-15-one (6 mg, 11.44 μmol, 6.00% yield) as a yellow solid.

Procedure for Preparation of Compound 017

Scheme 7

To a solution of 4-chloro-2-methylsulfanyl-pyrimidine-5-carbaldehyde (1 .2 g, 6.3 mmol, 1 eq) in MeCN (20 mL) was added DIEA (2.4 g, 19.0 mmol, 3.3 mL, 3 eq) to adjust pH=8-9 and 2-(3-aminopropoxy)ethanol (758.0 mg, 6.3 mmol, 1 eq). The mixture was stirred at 20°C for 4 hr. LC-MS showed desired compound was detected. The reaction mixture was filtered and concentrated under reduced pressure to remove solvent. The residue was purified by flash silica gel chromatography (20 g Silica Flash Column, Eluent of 0-52% Ethyl acetate/Petroleum ether gradient @ 100 mL/min) to give 4-[3-(2- hydroxyethoxy)propylamino]-2-methylsulfanyl-pyrimidine-5-carbaldehyde (1 g, 3.0 mmol, 48.0% yield, 83% purity) as yellow oil.

To a solution of 4-[3-(2-hydroxyethoxy)propylamino]-2-methylsulfanyl-pyrimidine-5- carbaldehyde (1 g, 3.6 mmol, 1 eq) in DMF (10 mL) was added Cs₂CO₃ (3.6 g, 11 .0 mmol, 3 eq) and tert-butyl 4-(2-ethoxy-2-oxo-ethyl)-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (1.2 g, 3.6 mmol, 1 eq). The mixture was stirred at 40°C for 12 hr. LC-MS showed desired compound was detected. The reaction mixture was diluted with H₂O (30 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with brine (20 mL x 3), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (20 g Silica Flash Column, Eluent of 0-45% Ethyl acetate/Petroleum ether gradient @ 100 mL/min) to give tert-butyl 4- [8-[3-(2-hydroxyethoxy)propyl]-2-methylsulfanyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl- 2, 3-dihydroquinoxaline-1 -carboxylate (736 mg, 1.0 mmol, 29.4% yield, 80% purity) as yellow oil.

At 0°C, a solution of tert-butyl 4-[8-[3-(2-hydroxyethoxy)propyl]-2-methylsulfanyl-7- oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (736 mg, 1.3 mmol, 1 eq) in DCM (5 mL) was added m-CPBA (331.0 mg, 1 .6 mmol, 85% purity, 1 .2 eq). The mixture was stirred at 20°C for 2 hr. LC-MS showed desired compound was detected. The reaction mixture was quenched by addition Na₂SO₃ (20 mL), and then extracted with dichloromethane (10 mL x 3). The combined organic layers were washed with NaHCO₃ (10 mL x 3), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give tertbutyl 4-[8-[3-(2-hydroxyethoxy)propyl]-2-methylsulfonyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8- methyl-2,3-dihydroquinoxaline-1 -carboxylate (670 mg, crude) as an orange solid.

To a solution of tert-butyl 4-[8-[3-(2-hydroxyethoxy)propyl]-2-methylsulfonyl-7-oxo- pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (670 mg, 1.1 mmol, 1 eq) in dioxane (6.7 mL) was added TFA (199.7 mg, 1.7 mmol, 130.1 μL, 1.5 eq) and 3-aminophenol (127.4 mg, 1.1 mmol, 1 eq). The mixture was stirred at 100°C for 2 hr. Then the mixture was replenished 3-aminophenol (127.4 mg, 1.1 mmol, 1 eq) and stirred at 100°C for 2 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by flash silica gel chromatography (20 g Silica Flash Column, Eluent of 0-60% Ethyl acetate/Petroleum ether gradient @ 100 mL/min) to give tert-butyl 4-[2-(3-hydroxyanilino)-8- [3-(2-hydroxyethoxy)propyl]-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3- dihydroquinoxaline-1-carboxylate (370 mg, 503.4 μmol, 43.1 % yield, 82% purity) as a yellow solid.

At 0°C, a mixture of tert-butyl 4-[2-(3-hydroxyanilino)-8-[3-(2-hydroxyethoxy)propyl]-7- oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (370 mg, 613.9 μmol, 1 eq), DTBAD (565.4 mg, 2.4 mmol, 4 eq), PPh₃ (644.0 mg, 2.4 mmol, 4 eq) in THF (18.5 mL) and Tol. (18.5 mL) was degassed and purged with N₂ for 3 times, and then the mixture was stirred at 20°C for 12 hr under N₂ atmosphere. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by flash silica gel chromatography (12 g Silica Flash Column, Eluent of 0-43% Ethyl acetate/Petroleum ether gradient @ 100 mL/min) to give tert-butyl 8-methyl-4-(16-oxo-8,11-dioxa-2,15,21 ,22- tetrazatetracyclo[13.6.2.13,7.019,23]tetracosa-1 (21 ),3,5,7(24),17,19,22-heptaen-17-yl)-2,3- dihydroquinoxaline-1 -carboxylate (70 mg, 96.3 μmol, 15.7% yield, 80.5% purity as a yellow solid.

A solution of tert-butyl 8-methyl-4-(16-oxo-8,11-dioxa-2,15,21 ,22- tetrazatetracyclo[13.6.2.13,7.019,23]tetracosa-1 (21 ),3,5,7(24),17,19,22-heptaen-17-yl)-2,3- dihydroquinoxaline-1 -carboxylate (70 mg, 119.7 μmol, 1 eq) in EtOAc (2.4 mL) and HCI/EtOAc (2 mL) 4 M was stirred at 20°C for 1 hr. LC-MS showed desired compound was detected.

The reaction mixture was concentrated under reduced pressure to give 17-(5-methyl-3,4- dihydro-2H-quinoxalin-1-yl)-8,11-dioxa-2,15,21 ,22- tetrazatetracyclo[13.6.2.13,7.019,23]tetracosa-1 (21 ),3,5,7(24),17,19,22-heptaen-16-one (90 mg, crude, HCI) as a yellow solid.

To a solution of 17-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-8,11-dioxa- 2,15,21 ,22- tetrazatetracyclo[13.6.2.13,7.019,23]tetracosa-1 (21 ),3,5,7(24),17,19,22-heptaen-16-one (10 mg, 19.1 μmol, 1 eq, HCI) in DCM (1 mL) was added dropwise TEA (5.8 mg, 57.5 μmol, 8.0 μL, 3 eq) and prop-2-enoyl chloride (1.7 mg, 19.1 μmol, 1.5 μL, 1 eq) in DCM (0.1 mL) at 0°C. The mixture was stirred at 0°C for 1 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under N₂ to remove solvent. The residue was purified by prep-HPLC (TFA conditioncolumn: Phenomenex luna C18 100x40mmx3 um; mobile phase: [H₂O(0.1 % TFA)-ACN];gradient:20%-60% B over 8.0 min), to give 17-(5- methyl-4-prop-2-enoyl-2,3-dihydroquinoxalin-1-yl)-8,11-dioxa-2, 15,21 ,22- tetrazatetracyclo[13.6.2.13,7.019,23]tetracosa-1 (21 ),3,5,7(24),17,19,22-heptaen-16-one (2.8 mg, 5.3 μmol, 27.9% yield, 100% purity) as a yellow solid.

Procedure for Preparation of Compound 006

Scheme 8

To a solution of 4-nitro-1 H-pyrazole (3 g, 26.5 mmol, 1 eq) and PTSA (456.9 mg, 2.7 mmol, 0.1 eq) in DCM (20 mL) was added 3,4-dihydro-2H-pyran (2.7 g, 31.8 mmol, 2.9 mL, 1 .2 eq) in DCM (10 mL) drop-wised. The mixture was stirred at 20°C for 12 hr. TLC showed desired compound was detected. The mixture was concentrated to get a residue and the residue was purified by column chromatography (SiO₂, Petroleum ether/Ethyl acetate = 50/1 to 25/1) to give 4-nitro-1-tetrahydropyran-2-yl-pyrazole (5 g, 25.4 mmol, 95.6% yield) as colorless oil.

To a mixture of Pd/C (1 g, 10% purity) in THF (60 mL) which degassed and purged with N₂ for 3 time was added 4-nitro-1-tetrahydropyran-2-yl-pyrazole (5 g, 25.4 mmol, 1 eq), and then the mixture was degassed and stirred at 25°C for 12 hr under H₂ atmosphere (15 psi). LC-MS showed desired compound was detected. The reaction mixture was filtered and the filtrate was concentrated to get 1-tetrahydropyran-2-ylpyrazol-4-amine (4 g, crude) as brown oil which was used into the next step without further purification.

To a solution of benzyl 4-(2-ethoxy-2-oxo-ethyl)-8-methyl-2,3-dihydroquinoxaline-1- carboxylate (1.1 g, 3.0 mmol, 1 eq) in DMF (15 mL) was added Cs₂CO₃ (2.9 g, 8.9 mmol, 3 eq) and 4-[3-(2-hydroxyethoxy)propylamino]-2-methylsulfanyl-pyrimidine-5-carbaldehyde (0.8 g, 3.0 mmol, 1 eq). The mixture was stirred at 40°C for 12 hr. LC-MS showed desired compound was detected. The reaction mixture was partitioned between brine (70 mL) and ethyl acetate (100 mL x 3). The organic phase was separated, washed with brine (50 mL x 2), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue, the residue was purified by flash silica gel chromatography (20 g Silica Flash Column, Eluent of 0~51 % Ethyl acetate / Petroleum ether gradient @ 100 mL/min) to give benzyl 4-[8-[3-(2- hydroxyethoxy)propyl]-2-methylsulfanyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3- dihydroquinoxaline-1 -carboxylate (2.2 g) as a red solid.

To a solution of benzyl 4-[8-[3-(2-hydroxyethoxy)propyl]-2-methylsulfanyl-7-oxo- pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (1.2 g, 2.1 mmol, 1 eq) in DCM (15 mL) was added m-CPBA (494.6 mg, 2.3 mmol, 80% purity, 1.1 eq). The mixture was stirred at 25°C for 1 hr. LC-MS showed desired compound was detected. The mixture was quenched by Sat. Na₂SO₃ 10 mL, then the mixture was extracted by DCM (10 mL x 3), the combined organic layers was washed by Sat. NaHCO₃ (10 mL x 3), then dried over Na₂SO₄, filtered and the filtrate was concentrated to get benzyl 4-[8-[3-(2- hydroxyethoxy)propyl]-2-methylsulfinyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3- dihydroquinoxaline-1-carboxylate (1.2 g, crude) as a red solid which was used into the next step without further purification.

To a solution of benzyl 4-[8-[3-(2-hydroxyethoxy)propyl]-2-methylsulfinyl-7-oxo- pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (1.2 g, 2.0 mmol, 1 eq) in IPA (20 mL) was added 1-tetrahydropyran-2-ylpyrazol-4-amine (339.1 mg, 2.0 mmol, 1 eq). The mixture was stirred at 80°C for 12 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated to give a residue and the residue was purified by flash silica gel chromatography (12 g Silica Flash Column, Eluent of 0-85% Ethyl acetate I Petroleum ether gradient @ 100 mL/min) to give benzyl 4-[8-[3-(2- hydroxyethoxy)propyl]-7-oxo-2-[(1-tetrahydropyran-2-ylpyrazol-4-yl)amino]pyrido[2,3- d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (1 g, 1.4 mmol, 70.9% yield) as a brown solid.

To a solution of benzyl 4-[8-[3-(2-hydroxyethoxy)propyl]-7-oxo-2-[(1-tetrahydropyran- 2-ylpyrazol-4-yl)amino]pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1- carboxylate (0.5 g, 719.7 μmol, 1 eq) in DCM (15 mL) was added TEA (364.1 mg, 3.6 mmol, 500.8 μL, 5 eq), TsCI (411.6 mg, 2.2 mmol, 3 eq) and DMAP (8.8 mg, 72.0 μmol, 0.1 eq). The mixture was stirred at 20°C for 12 hr. LC-MS showed desired compound was detected. The mixture was quenched by Sat. NaHCO₃ (10 mL) and then extracted by DCM (10 mL x 3), the organic layers was dried by Na₂SO₄, filtered and the filtrate was concentrated to get a residue. The residue was purified by flash silica gel chromatography (12 g Silica Flash Column, Eluent of 0-100% ethyl acetate I petroleum ether gradient @ 100 mL/min) to give benzyl 8-methyl-4-[7-oxo-8-[3-[2-(p-tolylsulfonyloxy)ethoxy]propyl]-2-[(1-tetrahydropyran-2- ylpyrazol-4-yl)amino]pyrido[2,3-d]pyrimidin-6-yl]-2,3-dihydroquinoxaline-1 -carboxylate (890 mg, 1.1 mmol, 72.8% yield) as a brown solid.

To a solution of benzyl 8-methyl-4-[7-oxo-8-[3-[2-(p-tolylsulfonyloxy)ethoxy]propyl]-2- [(1-tetrahydropyran-2-ylpyrazol-4-yl)amino]pyrido[2,3-d]pyrimidin-6-yl]-2,3- dihydroquinoxaline-1 -carboxylate (890 mg, 1.1 mmol, 1 eq) in EtOAc (10 mL) was added HCI/EtOAc (4 M, 1.6 mL, 6 eq). The mixture was stirred at 20°C for 1 hr. LC-MS showed desired compound was detected.

The mixture was concentrated to get a residue and the residue was triturated with petroleum ether I ethyl acetate = 3 / 1 , 40 mL at 20°C for 2 hr to give benzyl 8-methyl-4-[7-oxo-8-[3-[2- (p-tolylsulfonyloxy)ethoxy]propyl]-2-(1H-pyrazol-4-ylamino)pyrido[2,3-d]pyrimidin-6-yl]-2,3- dihydroquinoxaline-1 -carboxylate (870 mg crude, HCI) as a yellow solid.

To a solution of benzyl 8-methyl-4-[7-oxo-8-[3-[2-(p-tolylsulfonyloxy)ethoxy]propyl]-2- (1 H-pyrazol-4-ylamino)pyrido[2,3-d]pyrimidin-6-yl]-2,3-dihydroquinoxaline-1 -carboxylate (100 mg, 124.8 μmol, 1 eq, HCI) in DMF (15 mL) was added K₂CO₃ (86.2 mg, 624.0 μmol, 5 eq). The mixture was stirred at 80°C for 12 hr. LC-MS showed desired compound was detected. The mixture was poured into brine (15 mL) and then extracted by ethyl acetate (10 mL x 3), the combined organic layers were washed with brine (20 mL x 2), then dried by Na₂SO₄ filtered and the filtrate was concentrated to get a residue. The residue was purified by flash silica gel chromatography (4 g Silica Flash Column, Eluent of 0-75% ethyl acetate I petroleum ether gradient @ 100mL/min) to give benzyl 8-methyl-4-(14-oxo-9-oxa- 2,5,6, 13,19, 20-hexazatetracyclo[11.6.2.13,6.017,21]docosa-1(19),3(22),4,15,17,20-hexaen- 15-yl)-2,3-dihydroquinoxaline-1 -carboxylate (170 mg, 286.9 μmol, 57.5% yield) as yellow oil.

A mixture of Pd(OH)₂ (100 mg, 20% purity) in THF (10 mL) was degassed and purged with N₂ for 3 times, and then benzyl 8-methyl-4-(14-oxo-9-oxa-2,5,6, 13, 19,20- hexazatetracyclo[11 ,6.2.13,6.017,21]docosa-1 (19),3(22),4,15,17,20-hexaen-15-yl)-2,3- dihydroquinoxaline-1-carboxylate (170 mg, 286.9 μmol, 1 eq) was added, the mixture was stirred at 25°C for 12 hr under H₂ atmosphere (15 psi). LC-MS showed desired compound was detected. The mixture was filtered and the filtrate was concentrated to get 15-(5- methyl-3,4-dihydro-2H-quinoxalin-1-yl)-9-oxa-2,5,6,13,19,20- hexazatetracyclo[11 ,6.2.13,6.017,21]docosa-1 (19),3(22),4,15,17,20-hexaen-14-one (120 mg, crude) as brown oil which was used into the next step without further purification.

To a solution of 15-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-9-oxa-2,5,6,13,19,20- hexazatetracyclo[11 ,6.2.13,6.017,21]docosa-1 (19),3(22),4,15,17,20-hexaen-14-one (90 mg, 196.3 μmol, 1 eq) in DCM (3 mL) was added TEA (59.6 mg, 588.9 μmol, 82.0 μL, 3 eq) andprop-2-enoyl chloride (10.7 mg, 117.8 μmol, 9.6 μL, 0.6 eq). The mixture was stirred at 25°C for 1 hr. LC-MS showed desired compound was detected. The mixture was concentrated to get a residue. The residue was purified by prep-HPLC (TFA condition; column: Phenomenex luna C18 80 x 40mm x 3 um ;mobile phase: [H2O (0.1% TFA) - ACN]; gradient:25%-55% B over 8.0 min) to give 15-(5-methyl-4-prop-2-enoyl-2,3- dihydroquinoxalin-1-yl)-9-oxa-2,5,6,13,19,20-hexazatetracyclo[11.6.2.13,6.017,21]docosa- 1 (19),3(22),4,15,17,20-hexaen-14-one (9 mg, 16.6 μmol, 8.5% yield, 94.69% purity) as an orange solid. Procedure for Preparation of Compound 005

Scheme 9

A mixture of benzyl 4-[8-[3-[tert-butoxycarbonyl(methyl)amino]propyl]-2-[[1-(2,2- dimethoxyethyl)pyrazol-4- yl]amino]-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (1 .2 g, 1 .6 mmol, 1eq) in HCI (12 mL) and EtOAc (12 mL) was stirred at 20°C for 30 min.

LC-MS showed desired compound was detected. The mixture was concentrated to get a residue, the residue was purified by prep-HPLC (HCI condition column: Phenomenex Luna C18 75 x 30 mm x 3 um; mobile phase: [H₂O (0.04% HCI) - ACN]; gradient: 20%-50% B over 8.0 min.) to give benzyl 8-methyl-4-[8-[3-(methylamino)propyl]-7-oxo-2-[[1-(2- oxoethyl)pyrazol-4-yl]amino]pyrido[2,3-d]pyrimidin-6-yl]-2,3-dihydroquinoxaline-1 -carboxylate (420 mg, 675.6 μmol, 43.2% yield) as a yellow solid. To a solution of benzyl 8-methyl-4-[8-[3-(methylamino)propyl]-7-oxo-2-[[1-(2- oxoethyl)pyrazol-4-yl]amino]pyrido[2,3-d]pyrimidin-6-yl]-2,3-dihydroquinoxaline-1 -carboxylate (250 mg, 402.1 μmol, 1 eq) in MeOH (20 mL) which was adjust pH to 7-8 by TEA was adjust pH to 5-6 by AcOH, the mixture was stirred at 50°C for 12 hr, then NaBH₃CN (50.6 mg, 804.3 μmol, 2 eq) was added, the mixture was stirred at 20°C for 3 hr. LC-MS showed desired compound was detected. The mixture was concentrated to get a residue, the residue was purified by prep-HPLC (TFA condition; column: Phenomenex Luna C18 75 x 30 mm x 3 um; mobile phase: [H₂O (0.1% TFA) - ACN]; gradient: 25%-55% B over 8.0 min) to give benzyl 8-methyl-4-(9-methyl-14-oxo-2,5,6,9,13, 19,20- heptazatetracyclo[11 .6.2.13,6.017,21]docosa-1 (19), 3(22), 4, 15, 17,20-hexaen-15-y I )-2 , 3- dihydroquinoxaline-1 -carboxylate (35 mg, 57.8 μmol, 14.4% yield) as an orange solid.

A mixture of Pd(OH)₂ (50 mg, 20% purity) in THF (10 mL) was degassed and purged with N₂ for 3 times, and then benzyl 8-methyl-4-(9-methyl-14-oxo-2,5,6,9,13, 19,20- heptazatetracyclo[11 .6.2.13,6.017,21]docosa-1 (19), 3(22), 4, 15, 17,20-hexaen-15-y I )-2 , 3- dihydroquinoxaline-1 -carboxylate (35 mg, 57.8 μmol, 1 eq) was added, the mixture was stirred at 25°C for 12 hr under H₂ atmosphere (15 psi). LC-MS showed desired compound was detected. The mixture was filtered and the filtrate was concentrated to get 9-methyl-15- (5-methyl-3,4-dihydro-2H-quinoxalin-1 -yl)-2,5,6,9, 13,19,20- heptazatetracyclo[11 ,6.2.13,6.017,21]docosa-1 (19),3(22),4,15,17,20-hexaen-14-one (22 mg, crude) as yellow oil which was used into the next step without further purification.

To a solution of 9-methyl-15-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)- 2, 5, 6, 9, 13, 19,20-heptazatetracyclo[11 .6.2.13,6.017,21 ]docosa-1 (19), 3(22), 4, 15, 17,20- hexaen-14-one (22 mg, 46.7 μmol, 1 eq) in DCM (2 mL) was added TEA (14.2 mg, 140.0 μmol, 19.5 μL, 3 eq) and prop-2-enoyl chloride (3.0 mg, 32.7 μmol, 2.7 μL, 0.7 eq). The mixture was stirred at 20 °C for 1 hr. LC-MS showed desired compound was detected. The mixture was concentrated to get a residue. The residue was purified by prep-HPLC (TFA condition column: Phenomenex Luna C18 75 x 30 mm x 3 um; mobile phase: [H₂O (0.1% TFA) - ACN]; gradient: 10%-40% B over 8.0 min) to give 9-methyl-15-(5-methyl-4-prop-2- enoyl-2,3-dihydroquinoxalin-1-yl)-2,5,6,9,13,19,20- heptazatetracyclo[11 .6.2.13,6.017,21 ]docosa-1 ( 19), 3(22), 4, 15, 17,20-hexaen-14-one (8 mg, 12.5 μmol, 26.8% yield, 99.8% purity, TFA) as a brown solid. Procedure for Preparation of Compound 011 and Compound 013

Scheme 10

To a solution of 3-nitrophenol (5 g, 35.9 mmol, 7.1 mL, 1 eq) in MeCN (50 mL) was added K₂CO₃ (14.9 g, 107.8 mmol, 3 eq) and 3-bromoprop-1-ene (6.5 g, 53.9 mmol, 1.5 eq). The mixture was mixture was stirred at 0°C for 2 hr. Then the mixture was replenished 3- bromoprop-1-ene (4.3 g, 35.9 mmol, 1 q) and stirred at 40°C for 2 hr. TLC indicated 3- nitrophenol was consumed completely and one new spot formed. The reaction was clean according to TLC. The reaction mixture was filtered and concentrated under reduced pressure to remove solvent. The residue was purified by flash silica gel chromatography (40 g Silica Flash Column, Eluent of 0~3% Ethyl acetate/Petroleum ether gradient @ 100 mL/min) to give 1-allyloxy-3-nitro-benzene (6 g, 33.1 mmol, 92.2% yield, 99% purity) as yellow oil.

To a solution of 1-allyloxy-3-nitro-benzene (3 g, 16.7 mmol, 1 eq) in EtOH (30 mL) was added SnCl2.2H₂O (18.8 g, 83.7 mmol, 5 eq). The mixture was stirred at 75°C for 2 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was quenched with 40% NaOH to adjust pH=8-9, filtered and concentrated under reduced pressure to give 3-allyloxyaniline (2.2 g, crude) as yellow oil.

At 0°C, a solution of tert-butyl 8-methyl-4-(2-methylsulfanyl-7-oxo-8H-pyrido[2,3- d]pyrimidin-6-yl)-2,3-dihydroquinoxaline-1-carboxylate (500 mg, 1.1 mmol, 1 eq) in DCM (5 mL) was added m-CPBA (277.1 mg, 1.3 mmol, 85% purity, 1.2 eq). The mixture was stirred at 20°C for 2 hr. LC-MS showed desired compound was detected. The reaction mixture was quenched by addition Na₂SO₃ (20 mL), and then extracted with dichloromethane (10 mL x 3). The combined organic layers were washed with NaHCO₃ (10 mL x 3), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give tert-butyl 8-methyl-4-(2- methylsulfonyl-7-oxo-8H-pyrido[2,3-d]pyrimidin-6-yl)-2,3-dihydroquinoxaline-1 -carboxylate (530 mg, crude) as a orange solid.

To a solution of tert-butyl 8-methyl-4-(2-methylsulfonyl-7-oxo-8H-pyrido[2,3- d]pyrimidin-6-yl)-2,3-dihydroquinoxaline-1-carboxylate (380 mg, 805.8 μmol, 1 eq) in dioxane (3.8 mL) was added TFA (91 .8 mg, 805.8 μmol, 59.8 μL, 1 eq) and 3-allyloxyaniline (144.2 mg, 967.0 μmol, 1 .2 eq). The mixture was stirred at 80°C for 4 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The crude product was triturated with ethyl acetate (3 mL) at 20°C for 1 hr to give tert-butyl 4-[2-(3-allyloxyanilino)-7-oxo-8H-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3- dihydroquinoxaline-1-carboxylate (250 mg, 388.4 μmol, 48.2% yield, 84% purity) as a yellow solid.

To a solution of tert-butyl 4-[2-(3-allyloxyanilino)-7-oxo-8H-pyrido[2,3-d]pyrimidin-6- yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (200 mg, 369.9 μmol, 1 eq) in acetone (5 mL) was added Cs₂CO₃ (361 .6 mg, 1.1 mmol, 3 eq), Nal (11 .0 mg, 73.9 μmol, 0.2 eq) and 4- bromobut-1-ene (49.9 mg, 369.9 μmol, 37.5 μL, 1 eq). The mixture was stirred at 60°C for 12 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with H₂O (10 mL) and extracted with ethyl acetate (10 mL x 3). The combined organic layers were washed with brine (10 mL x 3), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiO₂, Petroleum ether/Ethyl acetate=1/1 ) to give tert-butyl 4-[2-(3-allyloxyanilino)-8-but-3-enyl-7-oxo- pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (125 mg, 191.2 μmol, 51.7% yield, 91 % purity) as a yellow solid.

A solution of tert-butyl 4-[2-(3-allyloxyanilino)-8-but-3-enyl-7-oxo-pyrido[2,3- d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (100 mg, 168.1 μmol, 1 eq) in Tol. (45 mL) was degassed and purged with N₂ for 3 times, and then the mixture was stirred at 110°C for 1 hr under N₂ atmosphere, benzylidene-[1 ,3-bis(2,4,6- trimethylphenyl)imidazolidin-2-ylidene]-dichloro-ruthenium;tricyclohexylphosphane (42.8 mg, 50.4 μmol, 0.3 eq) in Tol. (5 mL) was added and then the mixture was stirred at 110°C for 12 hr under N₂ atmosphere. LC-MS showed desired compound was detected.

The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (FA conditiomcolumn: Phenomenex luna C18 100x40mmx3 um; mobile phase: [H₂O(0.2% FA)-ACN];gradient:45%-75% B over 8.0 min) to give tert-butyl 8-methyl-4-[(10E)-15-oxo-8-oxa-2,14,20,21- tetrazatetracyclo[12.6.2.13,7.018,22]tricosa-1 (20),3,5,7(23),10,16,18,21-octaen-16-yl]-2,3- dihydroquinoxaline-1-carboxylate (20 mg, 33.8 μmol, 20.1 % yield, 96% purity) as a white solid and tert-butyl 8-methyl-4-[(10Z)-15-oxo-8-oxa-2,14,20,21- tetrazatetracyclo[12.6.2.13,7.018,22]tricosa-1 (20),3,5,7(23),10,16,18,21-octaen-16-yl]-2,3- dihydroquinoxaline-1-carboxylate (15 mg, 24.8 μmol, 14.8% yield, 94% purity) as a yellow solid.

A solution of tert-butyl 8-methyl-4-[(10E)-15-oxo-8-oxa-2,14,20,21- tetrazatetracyclo[12.6.2.13,7.018,22]tricosa-1 (20),3,5,7(23),10,16,18,21-octaen-16-yl]-2,3- dihydroquinoxaline-1 -carboxylate (20.0 mg, 35.3 μmol, 1 eq) in HCI/EtOAc (1 mL) 4 M and EtOAc (1 mL) was stirred at 20°C for 1 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to give (10E)-16-(5-methyl- 3,4-dihydro-2H-quinoxalin-1 -yl)-8-oxa-2, 14,20,21 - tetrazatetracyclo[12.6.2.13,7.018,22]tricosa-1 (20),3,5,7(23),10,16,18,21-octaen-15-one (22 mg, crude, HCI) as a yellow solid.

At 0°C, a solution of (10E)-16-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-8-oxa- 2,14,20,21-tetrazatetracyclo[12.6.2.13,7.018,22]tricosa-1 (20),3,5,7(23),10,16,18,21-octaen- 15-one (22 mg, 43.7 μmol, 1 eq, HCI) in THF (0.8 mL) and H₂O (0.2 mL) was added NaHCO₃ (11.0 mg, 131.2 μmol, 5.1 μL, 3 eq) and prop-2-enoyl chloride (3.9 mg, 43.7 μmol, 3.5 μL, 1 eq) in THF (0.1 mL). The mixture was stirred at 0°C for 30 min. Then the mixture was replenished prop-2-enoyl chloride (11.8 mg, 131.2 μmol, 10.6 μL, 3 eq) and stirred at 0°C for 30 min. LC-MS showed desired compound was detected. The reaction mixture was concentrated under N₂ to remove solvent. The residue was purified by prep-HPLC (FA condition column: Phenomenex luna C18 100x40mmx3 um; mobile phase: [H₂O(0.2% FA)- ACN];gradient:45%-80% B over 8.0 min) to give (10E)-16-(5-methyl-4-prop-2-enoyl-2,3- dihydroquinoxalin-1-yl)-8-oxa-2,14,20,21-tetrazatetracyclo[12.6.2.13,7.018,22]tricosa- 1 (20),3,5,7(23),10,16,18,21-octaen-15-one (Compound 011 ) (4.1 mg, 8.0 μmol, 18.3% yield, 100% purity) as a yellow solid.

A solution of tert-butyl 8-methyl-4-[(10Z)-15-oxo-8-oxa-2,14,20,21- tetrazatetracyclo[12.6.2.13,7.018,22]tricosa-1 (20),3,5,7(23),10,16,18,21-octaen-16-yl]-2,3- dihydroquinoxaline-1 -carboxylate (15 mg, 26.4 μmol, 1 eq) in HCI/EtOAc (1 mL) 4 M and EtOAc (1 mL) was stirred at 20°C for 1 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to give (10Z)-16-(5-methyl- 3,4-dihydro-2H-quinoxalin-1 -yl)-8-oxa-2, 14,20,21 - tetrazatetracyclo[12.6.2.13,7.018,22]tricosa-1 (20),3,5,7(23),10,16,18,21-octaen-15-one (18 mg, crude, HCI) as a yellow solid.

At 0°C, a solution of (10Z)-16-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-8-oxa- 2,14,20,21-tetrazatetracyclo[12.6.2.13,7.018,22]tricosa-1 (20),3,5,7(23),10,16,18,21-octaen- 15-one (18 mg, 35.7 μmol, 1 eq, HCI) in THF (0.8 mL) and H₂O (0.2 mL) was added NaHCO₃ (9.0 mg, 107.3 μmol, 4.1 μL, 3 eq) and prop-2-enoyl chloride (3.2 mg, 35.7 μmol, 2.9 μL, 1 eq) in TH F (0.1 mL). The mixture was stirred at 0°C for 30 min. Then the mixture was replenished prop-2-enoyl chloride (9.7 mg, 107.3 μmol, 8.7 μL, 3 eq) and stirred at 0°C for 30 min. LC-MS showed desired compound was detected. The reaction mixture was concentrated under N₂ to remove solvent. The residue was purified by prep-HPLC (FA condition column: Phenomenex luna C18100x40mmx3 um; mobile phase: [H₂O(0.2% FA)- ACN];gradient:45%-80% B over 8.0 min) to give (10Z)-16-(5-methyl-4-prop-2-enoyl-2,3- dihydroquinoxalin-1-yl)-8-oxa-2,14,20,21-tetrazatetracyclo[12.6.2.13,7.018,22]tricosa- 1(20),3,5,7(23),10,16,18,21-octaen-15-one (Compound 013) (1.9 mg, 3.7 μmol, 10.5% yield, 100% purity) as a yellow solid. 1H NMR (400 MHz, CHLOROFORM-d₆) δ = 8.55 (s, 1H), 8.08 (s, 1H), 7.57-7.52 (m,

1H), 7.47 (s, 1H), 7.22 (t, J=8.1 Hz, 1H), 6.96 (t, J =7.8 Hz, 1H), 6.74-6.66 (m, 2H), 6.58- 6.51 (m, 1H), 6.48 (d, J= 2.1 Hz, 1H), 6.45 - 6.34 (m, 2H), 5.82-5.73 (m, 1H), 5.71 -5.61 (m, 2H), 5.12 (dd, J= 5.0, 12.8 Hz, 1H), 4.74 (brs, 2H), 4.33 (brd, J= 7.0 Hz, 2H), 3.86 (dd, J= 3.9, 10.8 Hz, 1H), 3.62 (dt, J=5.8, 11.1 Hz, 1H), 3.19- 3.07 (m, 1H), 2.82 - 2.68 (m, 2H), 2.22 (s, 3H). MS (ESI): m/z = 521.4 [M+H]⁺

Procedure for Preparation of Compound 012

Scheme 11

To a solution of tert-butyl N-(3-hydroxyphenyl)carbamate (1 g, 4.7 mmol, 1 eq) in DMF (10 mL) was added K₂CO₃ (792.6 mg, 5.7 mmol, 1.2 eq) and 5-bromopent-1-ene (854.6 mg, 5.7 mmol, 678.3 μL, 1.2 eq). The mixture was stirred at 25°C for 12 hr. TLC indicated tert-butyl N-(3-hydroxyphenyl)carbamate was remained. 5-bromopent-1-ene (213.6 mg, 1 .4 mmol, 169.5 μL, 0.3 eq) was added and the mixture was stirred at 50°C for 12 hr. TLC indicated tert-butyl N-(3-hydroxyphenyl)carbamate was consumed completely. The residue was diluted with H₂O (50 mL) and extracted with ethyl acetate (50 mL x 3). The combined organic layers were washed with brine (40 mL x 3), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (12 g Silica Flash Column, Eluent of 0-100% Ethyl acetate/Petroleum ether gradient @ 100 mL/min) to give tert-butyl N-(3-pent-4- enoxyphenyl)carbamate (907.2 mg, 3.2 mmol, 68.4% yield) as a white solid.

A solution of tert-butyl N-(3-pent-4-enoxyphenyl)carbamate (400 mg, 1.4 mmol, 1 eq) in TFA (1 mL) and DCM (4 mL) was stirred at 25°C for 2 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to give 3-pent-4-enoxyaniline (615 mg, crude, TFA) as a brown oil.

To a solution of tert-butyl 8-methyl-4-(2-methylsulfinyl-7-oxo-8H-pyrido[2,3- d]pyrimidin-6-yl)-2,3-dihydroquinoxaline-1-carboxylate (317 mg, 695.8 μmol, 1 eq) in dioxane (5 mL) was added TFA (119.0 mg, 1 .0 mmol, 77.5 μL, 1 .5 eq) and 3-pent-4-enoxyaniline (304.0 mg, 1.0 mmol, 1.5 eq, TFA). The mixture was stirred at 100°C for 2 hr. 3-pent-4- enoxyaniline (101.3 mg, 347.9 μmol, 0.5 eq, TFA) was added, the mixture was stirred at 100°C for 2 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The crude product was triturated with ethyl acetate (5 ml) at 25°C for 1 hr to give tert-butyl 8-methyl-4-[7-oxo-2-(3-pent-4- enoxyanilino)-8H-pyrido[2,3-d]pyrimidin-6-yl]-2,3-dihydroquinoxaline-1 -carboxylate (180 mg, 227.9 μmol, 16.3% yield, 72% purity) as a yellow solid.

To a solution of tert-butyl 8-methyl-4-[7-oxo-2-(3-pent-4-enoxyanilino)-8H-pyrido[2,3- d]pyrimidin-6-yl]-2,3-dihydroquinoxaline-1 -carboxylate (170 mg, 298.9 μmol, 1 eq) in acetone (3 mL) was added Nal (8.9 mg, 59.7 μmol, 0.2 eq), Cs₂CO₃ (292.2 mg, 896.8 μmol, 3 eq) and 3-bromoprop-1-ene (36.1 mg, 298.9 μmol, 1 eq). The mixture was stirred at 60°C for 6 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with H₂O (20 mL) and extracted with ethyl acetate (10 mL x 3). The combined organic layers were washed with brine (10 mL x 2), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiO₂, Petroleum ether : Ethyl acetate = 1 :1 ) to give tert-butyl 4-[8-allyl-7-oxo-2-(3-pent-4-enoxyanilino)pyrido[2,3- d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (118 mg, 182.2 μmol, 60.9% yield, 94% purity) as a yellow solid.

To a solution of tert-butyl 4-[8-allyl-7-oxo-2-(3-pent-4-enoxyanilino)pyrido[2,3- d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (88 mg, 144.5 μmol, 1 eq) in Tol. (40 mL) was degassed and purged with N₂ for 3 times, and then the mixture was stirred at 110°C for 1 hr under N₂ atmosphere. The mixture was added benzylidene-[1 ,3-bis(2,4,6- trimethylphenyl)imidazolidin-2-ylidene]-dichloro-ruthenium;tricyclohexylphosphane (36.8 mg, 43.3 μmol, 0.3 eq) in Tol. (4 mL) and was stirred at 110°C for 21 hr under N₂ atmosphere. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-TLC (SiO₂, Petroleum ether : Ethyl acetate = 1 :1) to give tert-butyl 8-methyl-4-[(12Z)-16-oxo-8-oxa- 2,15,21 ,22-tetrazatetracyclo[13.6.2.13,7.019, 23]tetracosa1 (21 ), 3, 5, 7(24), 12, 17,19,22- octaen-17-yl]-2,3-dihydroquinoxaline-1-carboxylate (41.4 mg, 67.0 μmol, 46.3% yield, 94% purity) as a yellow solid.

A solution of tert-butyl 8-methyl-4-[(12Z)-16-oxo-8-oxa-2, 15, 21 ,22- tetrazatetracyclo[13.6.2.13,7.019, 23]tetracosa-1 (21 ), 3, 5, 7(24), 12, 17, 19,22-octaen-17-yl]-2, 3- dihydroquinoxaline-1-carboxylate (41.4 mg, 71.3 μmol, 1 eq) in HCI/EtOAc (1 mL, 4M) was stirred at 25°C for 0.5 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to give (12Z)-17-(5-methyl-3,4-dihydro- 2H-quinoxalin-1 -yl)-8-oxa-2, 15,21 ,22-tetrazatetracyclo[13.6.2.13,7.019, 23]tetracosa- 1 (21 ),3,5,7(24),12,17,19,22-octaen-16-one (40 mg, crude, HCI) as a yellow solid.

To a solution of (12Z)-17-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-8-oxa-2,15,21 ,22- tetrazatetracyclo[13.6.2.13,7.019, 23]tetracosa-1 (21 ), 3, 5, 7(24), 12, 17, 19,22-octaen-16-one (30 mg, 58.0 μmol, 1 eq, HCI) in THF (0.8 mL) and H₂O (0.2 mL) was added NaHCO₃ (14.6 mg, 174.0 μmol, 6.7 μL, 3 eq). The mixture was added dropwise prop-2-enoyl chloride (5.2 mg, 58.0 μmol, 4.7 μL, 1 eq) in THF (0.1 mL) at 0°C and stirred at 0°C for 0.5 hr. LC-MS showed (12Z)-17-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-8-oxa-2, 15,21 ,22- tetrazatetracyclo[13.6.2.13,7.019, 23]tetracosa-1 (21 ), 3, 5, 7(24), 12, 17, 19,22-octaen-16-one remained. The mixture was added dropwise prop-2-enoyl chloride (2.6 mg, 29.0 μmol, 2.3 μL, 0.5 eq) in THF (0.1 mL) at 0°C and stirred at 0°C for 0.5 hr. LC-MS showed (12Z)-17-(5- methyl-3,4-dihydro-2H-quinoxalin-1-yl)-8-oxa-2, 15, 21 ,22- tetrazatetracyclo[13.6.2.13,7.019, 23]tetracosa-1 (21 ), 3, 5, 7(24), 12, 17, 19,22-octaen-16-one remained. The mixture was added dropwise prop-2-enoyl chloride (1.5 mg, 17.4 μmol, 1.4 μL, 0.3 eq) in THF (0.1 mL) at 0°C and stirred at 0°C for 0.5 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (TFA condition; column: Phenomenex luna C18 100x40mmx3 um; mobile phase: [H₂O(0.1% TFA)- ACN];gradient:30%-75% B over 8.0 min) to give (12Z)-17-(5-methyl-4-prop-2-enoyl-2,3- dihydroquinoxalin-1 -yl)-8-oxa-2, 15,21 ,22-tetrazatetracyclo[13.6.2.13,7.019,23]tetracosa- 1 (21 ),3,5,7(24),12,17,19,22-octaen-16-one (7 mg, 13.0 μmol, 22.5% yield, 100% purity) as a yellow solid. Procedure for Preparation of Compound 015

Scheme 12

To a solution of 3-nitrophenol (1 g, 7.2 mmol, 1 .4 mL, 1 eq) in DMF (20 mL) was added 3-bromo-1 ,1 -dimethoxy-propane (2.6 g, 14.4 mmol, 2.0 mL, 2 eq) and K₂CO₃ (3.0 g, 21.6 mmol, 3 eq). The mixture was stirred at 80°C for 4 hr. TLC indicated one new spot formed.

The residue was diluted with H₂O (80 mL) and extracted with ethyl acetate (100 mL x 4). The combined organic layers were washed with brine (100 mL x 3), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (12 g Silica Flash Column, Eluent of 0-1 % Ethyl acetate/Petroleum ether gradient @ 80 mL/min) to give 1-(3,3-dimethoxypropoxy)-3-nitro- benzene (1.7 g, 7.1 mmol, 98.0% yield) as a light yellow oil.

To a solution of Pd/C (1 g, 10% purity) in MeOH (20 mL) was added 1 -(3,3- dimethoxypropoxy)-3-nitro-benzene (1.7 g, 7.1 mmol, 1 eq) under N₂ atmosphere. The suspension was degassed and purged with H₂ for 3 times. The mixture was stirred under H₂ (15 Psi) at 20 °C for 2 hr. LC-MS showed desired compound was detected. The reaction mixture filtered and concentrated under reduced pressure to give 3-(3,3- dimethoxypropoxy)aniline (1.61 g, crude) as a black oil.

To a solution of benzyl 4-(2-ethoxy-2-oxo-ethyl)-8-methyl-2,3-dihydroquinoxaline-1- carboxylate (5.2 g, 14.1 mmol, 1 eq) in DMF (200 mL) was added 4-amino-2-methylsulfanyl- pyrimidine-5-carbaldehyde (2.4 g, 14.1 mmol, 1 eq) and Cs₂CO₃ (13.7 g, 42.2 mmol, 3 eq). The mixture was stirred at 60°C for 12 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with H₂O (800 mL) and extracted with ethyl acetate (800 mL x 4). The combined organic layers were washed with brine (800 mL x 3), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (220 g Silica Flash Column, Eluent of 0-45% Ethyl acetate/Petroleum ether gradient @ 150 mL/min). Then the product was triturated with methyl tertiary ether at 25°C for 30 min. The reaction mixture filtered and concentrated under reduced pressure to give benzyl 8-methyl-4-(2-methylsulfanyl-7-oxo-8H-pyrido[2,3- d]pyrimidin-6-yl)-2,3-dihydroquinoxaline-1-carboxylate (20.6 g, 43.5 mmol, 61.9% yield) as a yellow solid.

To a solution of benzyl 8-methyl-4-(2-methylsulfanyl-7-oxo-8H-pyrido[2,3-d]pyrimidin- 6-yl)-2,3-dihydroquinoxaline-1-carboxylate (6 g, 12.7 mmol, 1 eq) in DMF (60 mL) was added tert-butyl 2, 2-dioxooxathiazolidine-3-carboxylate (3.1 g, 13.9 mmol, 1.1 eq), Cs₂CO₃ (12.38 g, 38.01 mmol, 3 eq) and Nal (379.8 mg, 2.5 mmol, 0.2 eq). The mixture was stirred at 100°C for 12 hr. LC-MS showed 21.6% Reactant 1 was remained. Then the mixture was added tert-butyl 2, 2-dioxooxathiazolidine-3-carboxylate (1.4 g, 6.3 mmol, 0.5 eq), the mixture was stirred at 120°C for 2 hr. The mixture was stirred at 25°C for 12 hr. The residue was diluted with H₂O (200 mL) and extracted with ethyl acetate (300 mL x 4). The combined organic layers were washed with brine (300 mL x 3), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (40 g Silica Flash Column, Eluent of 0-31% Ethyl acetate/Petroleum ether gradient @ 120 mL/min) to give benzyl 4-[8-[2-(tert- butoxycarbonylamino)ethyl]-2-methylsulfanyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3- dihydroquinoxaline-1 -carboxylate (7.2 g, 11.6 mmol, 91 .9% yield) as a yellow oil.

To a solution of benzyl 4-[8-[2-(tert-butoxycarbonylamino)ethyl]-2-methylsulfanyl-7- oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (3.6 g, 5.8 mmol, 1 eq) in THF (100 mL) was added NaH (349.2 mg, 8.7 mmol, 60% purity, 1 .5 eq) at 0°C. The mixture was stirred at 0°C for 1 hr. Then the mixture was added CH₃I (1 .2 g, 8.7 mmol, 543.6 μL, 1 .5 eq) in THF (1 mL) at 0°C under N₂ atmosphere. The mixture was stirred at 20°C for 12 hr. LC-MS showed desired compound was detected. The reaction mixture was quenched by addition NH₄CI (100 mL) at 0 °C, and extracted with ethyl acetate (200 mL x 4). The combined organic layers were washed with brine (100 mL x 3), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (40 g Silica Flash Column, Eluent of 0-23% Ethyl acetate/Petroleum ether gradient @ 120 mL/min) to give benzyl 4-[8-[2-[tert- butoxycarbonyl(methyl)amino]ethyl]-2-methylsulfanyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8- methyl-2,3-dihydroquinoxaline-1 -carboxylate (6.0 g, 9.5 mmol, 82.0% yield) as a yellow oil.

To a solution of benzyl 4-[8-[2-[tert-butoxycarbonyl(methyl)amino]ethyl]-2- methylsulfanyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1- carboxylate (3 g, 4.8 mmol, 1 eq) in DCM (60 mL) was added m-CPBA (1.5 g, 7.1 mmol, 85% purity, 1 .5 eq) at 0°C. The mixture was stirred at 20°C for 1 hr. The residue was diluted with Na₂SO₃ (50 mL) and extracted with dichloromethane (60 mL x 4). The combined organic layers were washed with NaHCO₃ (60 mL x 3), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give benzyl 4-[8-[2-[tert- butoxycarbonyl(methyl)amino]ethyl]-2-methylsulfonyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8- methyl-2,3-dihydroquinoxaline-1 -carboxylate (3.1 g, crude) as an orange solid.

To a solution of 3-(3,3-dimethoxypropoxy)aniline (956.3 mg, 4.5 mmol, 3 eq) in THF (30 mL) was added LiHMDS (1 M, 7.5 mL, 5 eq) under N₂ atmosphere, the mixture was stirred at -78°C for 15 min under N₂ atmosphere. Then the mixture was added benzyl 4-[8- [2-[tert-butoxycarbonyl(methyl)amino]ethyl]-2-methylsulfonyl-7-oxo-pyrido[2,3-d]pyrimidin-6- yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (1 g, 1.5 mmol, 1 eq) in THF (8 mL), the mixture was stirred at -78°C for 0.5 hr under N₂ atmosphere. The mixture was stirred at 25°C for 1 hr. LC-MS showed desired compound was detected. The residue was diluted with NH₄CI (40 mL) and extracted with ethyl acetate (60 mL x 4). The combined organic layers were washed with brine (60 mL x 1 ), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (20 g Silica Flash Column, Eluent of 0-38% Ethyl acetate/Petroleum ether gradient @ 80 mL/min) to give benzyl 4-[8-[2-[tert-butoxycarbonyl(methyl)amino]ethyl]-2-[3- (3,3-dimethoxypropoxy)anilino]-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3- dihydroquinoxaline-1-carboxylate (485 mg, 610.9 μmol, 40.5% yield) as a yellow solid.

A mixture of benzyl 4-[8-[2-[tert-butoxycarbonyl(methyl)amino]ethyl]-2-[3-(3,3- dimethoxypropoxy)anilino]-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3- dihydroquinoxaline-1-carboxylate (485 mg, 610.9 μmol, 1 eq) in HCI (5 mL, 4M) and EtOAc (5 mL) was stirred at 20°C for 12 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent by lyophilization to give benzyl 8-methyl-4-[(11 E)-12-methyl-16-oxo-8-oxa-2, 15,21 , 22-tetraza- 12-azoniatetracyclo[13.6.2.13,7.019, 23]tetracosa-1 (21 ), 3, 5, 7(24), 11 ,17,19,22-octaen-17-yl]- 2, 3-dihydroquinoxaline-1 -carboxylate (480 mg, crude) as a yellow solid.

To a solution of Pd/C (480 mg, 10% purity) in MeOH (7 mL) was added benzyl 8- methyl-4-[(11 E)-12-methyl-16-oxo-8-oxa-2,15,21 ,22-tetraza-12- azoniatetracyclo[13.6.2.13,7.019,23]tetracosa-1 (21 ), 3, 5, 7(24), 11 ,17,19,22-octaen-17-y l]-2, 3- dihydroquinoxaline-1-carboxylate (480 mg, 761.0 μmol, 1 eq)and TEA (153.9 mg, 1522.1 μmol, 211 .9 μL, 2 eq) under N₂ atmosphere. The suspension was degassed and purged with H₂ for 3 times. The mixture was stirred under H₂ (15 Psi) at 20 °C for 1 hr. LC-MS showed desired compound was detected. The reaction mixture filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (TFA condition column: Phenomenex luna C18 80 x 30 mm x 3 um; mobile phase: [H₂O (0.1% TFA)-ACN]; gradient: 10%-40% B over 8.0 min) to give 12-methyl-17-(5-methyl-3,4-dihydro- 2H-quinoxalin-1 -yl)-8-oxa-2, 12, 15,21 ,22-pentazatetracyclo[13.6.2.13,7.019, 23]tetracosa- 1 (21 ),3,5,7(24),17,19,22-heptaen-16-one (7 mg, 14.07 μmol, 1 .4% yield) as a yellow solid.

To a solution of 12-methyl-17-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-8-oxa- 2,12,15,21 ,22-pentazatetracyclo[13.6.2.13,7.019, 23]tetracosa-1 (21 ), 3, 5, 7(24), 17, 19,22- heptaen-16-one (7 mg, 14.1 μmol, 1 eq) in THF (0.6 mL) and H₂O (0.2 mL) was added NaHCO₃ (3.6 mg, 42.2 μmol, 1.6 μL, 3 eq) and prop-2-enoyl chloride (636.6 μg, 7.0 μmol, 5.71e-1 μL, 0.5 eq) at 0°C. The mixture was stirred at 0°C for 0.5 hr. LC-MS showed 36.2% Reactant 1 was remained, then the mixture was added prop-2-enoyl chloride (636.6 μg, 7.0 μmol, 5.71e-1 μL, 0.5 eq) at 0°C. The mixture was stirred at 0°C for 0.5 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (TFA condition column: Phenomenex luna C18 100 x 40 mm x 3 um; mobile phase: [H₂O (0.1% TFA)-ACN]; gradient:20%-50% B over 8.0 min) to give 12-methyl-17-(5-methyl-4-prop-2-enoyl-2,3- dihydroquinoxalin-1 -yl)-8-oxa-2, 12,15,21 ,22-pentazatetracyclo[13.6.2.13,7.019,23]tetracosa-

1 (21 ),3,5,7(24),17,19,22-heptaen-16-one (1.42 mg, 2.51 μmol, 17.88% yield, 97.7% purity) as a yellow solid.

VT ¹H NMR (400 MHz, DMSO-d₆) δ = 9.50 (br s, 1 H), 8.64 (s, 1 H), 7.42 (s, 1 H), 7.25 - 7.15 (m, 2H), 6.99 (d, J = 8.3 Hz, 1 H), 6.57 - 6.44 (m, 2H), 6.41 - 6.33 (m, 1 H), 6.32 - 6.24 (m, 1 H), 5.98 - 5.83 (m, 1 H), 5.60 - 5.28 (m, 1 H), 4.55 (br t, J = 6.1 Hz, 2H), 4.17 - 4.07 (m,

2H), 3.75 (br d, J = 4.9 Hz, 2H), 3.52 - 3.49 (m, 2H), 3.39 - 3.35 (m, 2H), 3.05 - 2.88 (m, 3H), 2.71 (t, J = 6.3 Hz, 2H), 2.10 (s, 3H), 1.98 - 1.90 (m, 2H).

Procedure for Preparation of Compound 014. Compound 019. and Compound 020

Scheme 13

To a solution of 6-bromohexan-2-one (2.8 g, 15.6 mmol, 1 eq) in MeOH (30 mL) was added NaBH₄ (890 mg, 23.5 mmol, 1 .5 eq) at 0°C. The mixture was stirred at 0°C for 1 hr. TLC indicated Reactant 1 was consumed completely and new spot formed. The reaction mixture was quenched by addition NH₄CI (50 mL) at 0°C, and then diluted with H₂O (20 mL) and ethyl acetate (100 mL x 3). The organic phase was separated washed with brine (50 mL x 1), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (40 g Silica Flash Column, Eluent of 0-30% Ethyl acetate/Petroleum ether gradient @ 150 mL/min) to give 6- bromohexan-2-ol (3 g) as an off-white oil.

To a solution of tert-butyl 8-methyl-4-(2-methylsulfanyl-7-oxo-8H-pyrido[2,3- d]pyrimidin-6-yl)-2,3-dihydroquinoxaline-1-carboxylate (2 g, 4.5 mmol, 1 eq) and 6- bromohexan-2-ol (1.1 g, 5.9 mmol, 1 .3 eq) in acetone (30 mL) was added Nal (136.4 mg, 910.0 μmol, 0.2 eq) and Cs₂CO₃ (4.4 g, 13.6 mmol, 3 eq). The mixture was stirred at 60°C for 12 hr. LC-MS showed the desired compound was detected. The reaction mixture was partitioned between H₂O (100 mL) and ethyl acetate (100 mL x 3). The organic phase was separated washed with brine (50 mL x 1 ), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (40 g Silica Flash Column, Eluent of 0-40% Ethyl acetate/Petroleum ether gradient @ 150 mL/min) to give tert-butyl 4-[8-(5-hydroxyhexyl)-2-methylsulfanyl-7-oxo- pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (2.4 g, 4.4 mmol, 97.7% yield) as a white solid.

To a solution of tert-butyl 4-[8-(5-hydroxyhexyl)-2-methylsulfanyl-7-oxo-pyrido[2,3- d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (1.9 g, 3.5 mmol, 1 eq) in DCM (30 mL) was added m-CPBA (1.0 g, 5.2 mmol, 85% purity, 1 .5 eq) at 0°C. The mixture was stirred at 25°C for 2 hr. LC-MS showed the desired compound was detected. The reaction mixture was partitioned between Na₂SO₃ (100 mL) and dichloromethane (100 mL x 3). The organic phase was separated washed with NaHCO₃ (50 mL x 3), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give tert-butyl 4-[8-(5- hydroxyhexyl)-2-methylsulfonyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3- dihydroquinoxaline-1 -carboxylate (2.4 g, crude) as a yellow solid.

To a solution of 2-fluoropyridin-4-amine (215.7 mg, 1.9 mmol, 2 eq) in THF (16 mL) was added LiHMDS (1 M, 3.85 mL, 4 eq) under N₂ atmosphere, the mixture was stirred at - 78°C for 30 min under N₂ atmosphere. Then the mixture was added tert-butyl 4-[8-(5- hydroxyhexyl)-2-methylsulfonyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3- dihydroquinoxaline-1-carboxylate (550 mg, 962.0 μmol, 1 eq) in THF (10 mL), the mixture was stirred at 25°C for 12 hr under N₂ atmosphere. LC-MS showed the desired compound was detected. The reaction mixture was quenched by addition NH₄CI (50 mL), and then extracted with ethyl acetate (60 mL x 3). The organic phase was separated washed with brine (30 mL x 1 ), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (20 g Silica Flash Column, Eluent of 0-50% Ethyl acetate/Petroleum ether gradient @ 120 mL/min) to give tert-butyl 4-[2-[(2-fluoro-4-pyridyl)amino]-8-(5-hydroxyhexyl)-7-oxo-pyrido[2,3- d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (970 mg) as a yellow solid.

To a solution of tert-butyl 4-[2-[(2-fluoro-4-pyridyl)amino]-8-(5-hydroxyhexyl)-7-oxo- pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (970 mg, 1.6 mmol, 1 eq) in DMF (100 mL) was added NaH (446.2 mg, 1 1.1 mmol, 60% purity, 6.9 eq). The mixture was stirred at 50°C for 12 hr under N₂. LC-MS showed the desired compound was detected.

The reaction mixture was partitioned between NH₄CI (100 mL) and ethyl acetate (150 mL x 3). The organic phase was separated washed with brine (60 mL x 4), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (20 g Silica Flash Column, Eluent of 0-60% Ethyl acetate/Petroleum ether gradient @ 100 mL/min) to give tert-butyl 8-methyl-4-(9-methyl-15- oxo-8-oxa-2,6,14,20,21-pentazatetracyclo[12.6.2.1^3,7.0^18,22]tricosa-1 (20),3,5,7(23), 16,18,21- heptaen-16-yl)-2,3-dihydroquinoxaline-1 -carboxylate (640 mg, 1.1 mmol, 68.2% yield) as a yellow solid.

To a solution of tert-butyl 8-methyl-4-(9-methyl-15-oxo-8-oxa-2,6,14,20,21- pentazatetracyclo[12.6.2.13,7.018,22]tricosa-1 (20),3,5,7(23),16, 18,21-heptaen-16-yl)-2,3- dihydroquinoxaline-1-carboxylate (320 mg, 548.25 μmol, 1 eq) in EtOAc (2 mL) was added HCI/EtOAc (2 mL, 4 M). The mixture was stirred at 25°C for 2 hr. LC-MS showed the desired compound was detected. The reaction mixture was concentrated under reduced pressure to give 9-methyl-16-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-8-oxa-2,6, 14,20,21- pentazatetracyclo[12.6.2.13,7.018,22]tricosa-1 (20),3,5,7(23),16, 18,21-heptaen-15-one (290 mg, crude) as a yellow solid.

To a solution of 9-methyl-16-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-8-oxa-

2,6, 14,20,21 -pentazatetracyclo[12.6.2.1^3,7.0^18,22]tricosa-1 (20), 3, 5, 7(23), 16,18,21 -heptaen-15- one (130 mg, 250.0 μmol, 1 eq, HCI) in THF (3 mL) and H₂O (1 mL) was added NaHCO₃ (150 mg, 1 .8 mmol, 69.4 μL, 7.14 eq) and prop-2-enoyl chloride (22.6 mg, 250.0 μmol, 20.3 μL, 1 eq) at 0°C. The mixture was stirred at 0°C for 1 hr. LC-MS showed the desired compound was detected. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (FA condition, column: Phenomenex Luna C18 100 x 30 mm x 5 um; mobile phase: [H₂O(0.2% FA)-ACN]; gradient: 15%-45% B over 8.0 min) to give 9-methyl-16-(5-methyl-4-prop-2-enoyl-2,3-dihydroquinoxalin-1-yl)-8- oxa-2,6, 14,20,21-pentazatetracyclo[12.6.2.1^3,7.0^18,22]tricosa-1 (20),3,5,7(23), 16,18, 21 -heptaen-15-one (Compound 014) (6.2 mg, 11.09 μmol, 2.22% yield, 96.15% purity) as a yellow solid. The residue was separated by SFC [column: DAICEL CHIRALPAK AD (250 mm x 30 mm, 10 um); mobile phase: [CO₂-EtOH (0.1% NH₃H₂O)]; B%:40%, isocratic elution mode] to give (9S)-9-methyl-16-(5-methyl-4-prop-2-enoyl-2,3-dihydroquinoxalin-1-yl)-8-oxa- 2,6, 14,20,21 -pentazatetracyclo[12.6.2.1^3,7.0^18,22]tricosa-1 (20), 3, 5, 7(23), 16,18,21 -heptaen-15- one (Compound 019) (25.8 mg, 47.8 μmol, 19.1% yield, 99.5% purity) and (9R)-9-methyl- 16-(5-methyl-4-prop-2-enoyl-2,3-dihydroquinoxalin-1-yl)-8-oxa-2,6,14,20,21- pentazatetracyclo[12.6.2.1^3,7.0^18,22]tricosa-1(20),3,5,7(23),16,18,21-heptaen-15-one (Compound 020) (17.3 mg, 32.1 μmol, 12.8% yield, 99.3% purity) as a yellow solid. Compound 014: ¹H NMR (400 MHz, DMSO-d₆)δ = 10.57 (d, J= 1.8 Hz, 1H), 8.86

(s, 1H), 8.05-7.91 (m,3H), 7.03-6.91 (m, 1H), 6.80-6.75 (m, 1H), 6.69 (d, J=7.4 Hz, 1H), 6.42-6.21 (m, 3H), 5.84-5.69 (m, 1H), 4.91 (brdd, J= 5.3, 12.8 Hz, 1H), 4.54-4.41 (m, 2H), 4.14 - 3.91 (m, 1H), 3.85 - 3.70 (m, 1H), 3.52 - 3.39 (m, 1H), 3.04 (dt, J = 4.8, 12.0 Hz, 1H), 2.13 (s, 4H), 1.97- 1.82 (m, 1H), 1.79- 1.66 (m, 1H), 1.61 - 1.45 (m, 3H), 1.38- 1.31 (m,3H). MS (ESI): m/z = 538.4 [M+H]⁺

Procedure for Preparation of Compound 008

Scheme 14

To a solution of tert-butyl 8-methyl-4-(2-methylsulfanyl-7-oxo-8H-pyrido[2,3- d]pyrimidin-6-yl)-2,3-dihydroquinoxaline-1-carboxylate (5 g, 11.4 mmol, 1 eq)and 5- bromopentan-1-ol (2.9 g, 17.1 mmol, 1 .5 eq) in acetone (60 mL) was added Nal (170.5 mg, 1.1 mmol, 0.1 eq) and Cs₂CO₃ (11.1 g, 34.1 mmol, 3 eq). The mixture was stirred at 60°C for 4 hr. LC-MS showed desired mass was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with H₂O 50 mL and extracted with ethyl acetate (50 mL x 4). The combined organic layers were washed with brine(50 mL x 1), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (20 g Silica Flash Column, Eluent of 0-30% ethyl acetate I petroleum ether gradient @ 100 mL/min) to give tert-butyl 4-[8-(5-hydroxypentyl)-2-methylsulfanyl-7-oxo-pyrido[2,3- d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (3 g, 5.7 mmol, 50.2% yield) as a yellow solid.

To a solution of tert-butyl 4-[8-(5-hydroxypentyl)-2-methylsulfanyl-7-oxo-pyrido[2,3- d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (3 g, 5.7 mmol, 1 eq) in DCM (30 mL) was added m-CPBA (1.4 g, 6.9 mmol, 85% purity, 1 .2 eq). The mixture was stirred at 25°C for 2 hr. LC-MS showed desired compound was detected. The residue was diluted with Na₂SO₃ 50 mL and extracted with dichloromethane (50 mL x 3). The combined organic layers were washed with NaHCO₃ (50 mL x 3), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (20 g Silica Flash Column, Eluent of 0-100% ethyl acetate I petroleum ether gradient @ 100mL/min) to give tert-butyl 4-[8-(5-hydroxypentyl)-2- methylsulfinyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1- carboxylate (2.3 g, crude) as a yellow solid.

To a solution of tert-butyl 4-[8-(5-hydroxypentyl)-2-methylsulfinyl-7-oxo-pyrido[2,3- d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (200 mg, 369.2 μmol, 1 eq) and 5-aminobenzene-1 ,3-diol (55.4 mg, 443.1 μmol, 1.2 eq) in dioxane (2 mL) was added TFA (63.2 mg, 553.9 μmol, 41 .1 μL, 1.5 eq). The mixture was stirred at 100°C for 1 hr. LC- MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with NaHCO₃ 50 mL and extracted with ethyl acetate (50 mL x 4). The combined organic layers were washed with brine (50 mL x 1 ), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (20 g Silica Flash Column, Eluent of 0-70% ethyl acetate I petroleum ether gradient @ 100 mL/min) to give tert-butyl 4-[2-(3,5-dihydroxyanilino)-8-(5-hydroxypentyl)-7-oxo-pyrido[2,3-d]pyrimidin-6- yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (100 mg, 165.9 μmol, 6.4% yield) as a yellow solid.

To a solution of tert-butyl 4-[2-(3,5-dihydroxyanilino)-8-(5-hydroxypentyl)-7-oxo- pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (100 mg, 165.9 μmol, 1 eq) and DTBAD (152.8 mg, 663.7 μmol, 4 eq) in THF (5 mL) and Tol. (5 mL) was added PPh₃ (174.1 mg, 663.7 μmol, 4 eq). The mixture was stirred at 70°C for 12 hr under N₂. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with H₂O 50 mL and extracted with ethyl acetate (50 mL x 4). The combined organic layers were washed with brine (50 mL x 1 ), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiO₂, Petroleum ether : Ethyl acetate= 1 :1 ) to give tert-butyl 4-(5-hydroxy-15-oxo-8-oxa-2, 14,20,21 - tetrazatetracyclo [12.6.2.1^3,7.0^18,22]tricosa-1 (20), 3, 5, 7(23), 16,18,21 -heptaen-16-yl )-8-methyl-

2,3-dihydroquinoxaline-1-carboxylate (90 mg, 61.6 μmol, 37.1 % yield, 40% purity) as a yellow solid.

To a solution of tert-butyl 4-(5-hydroxy-15-oxo-8-oxa-2, 14,20,21 - tetrazatetracyclo [12.6.2.1^3,7.0^18,22]tricosa-1 (20), 3, 5, 7(23), 16,18,21 -heptaen-16-yl )-8-methyl-

2,3-dihydroquinoxaline-1-carboxylate (90 mg, 61.6 μmol, 1 eq) and 2-bromo-N,N-dimethyl- ethanamine (14.3 mg, 61.6 μmol, 1 eq, HBr) in DMF (1 mL) was added Cs₂CO₃ (68.1 mg, 492.6 μmol, 8 eq). The mixture was stirred at 70°C for 12 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC TFA condition column: Phenomenex Luna C18 75 x 30 mm x 3 um; mobile phase: [H₂O (0.1 % TFA) - ACN]; gradient: 35%-65% B over 8.0 min to give tert-butyl 4-[5-[2-(dimethylamino)ethoxy]-15-oxo- 8-oxa-2, 14,20,21 -tetrazatetracyclo[12.6.2. 1^3,7.0^18,22]tricosa-1 (20), 3, 5, 7(23), 16, 18,21- heptaen-16-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (18 mg, 27.5 μmol, 44.6% yield) as a yellow solid.

A mixture of tert-butyl 4-[5-[2-(dimethylamino)ethoxy]-15-oxo-8-oxa-2, 14,20,21 - tetrazatetracyclo [12.6.2.1^3,7.0^18,22]tricosa-1 (20), 3, 5, 7(23), 16,18,21 -heptaen-16-yl]-8-methyl-

2, 3-dihydroquinoxaline-1 -carboxylate (25 mg, 38.1 μmol, 1 eq) in EtOAc (0.5 mL) and HCI/EtOAc (0.5 mL,4M ) was stirred at 25°C for 1 hr. LC-MS showed desired compound was detected.

The reaction mixture was concentrated under reduced pressure to remove solvent to give 5- [2-(dimethylamino)ethoxy]-16-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-8-oxa-2, 14,20,21- tetrazatetracyclo[12.6.2.1^3,7.0^18,22]tricosa-1 (20),3,5,7(23),16,18,21-heptaen-15-one (21 mg, crude) as a white solid.

To a solution of 5-[2-(dimethylamino)ethoxy]-16-(5-methyl-3,4-dihydro-2H-quinoxalin- 1 -yl)-8-oxa-2, 14,20,21 -tetrazatetracyclo[12.6.2.1^3,7.0^18,22]tricosa-1 (20), 3, 5, 7(23), 16, 18,21- heptaen-15-one (21 mg, 37.79 μmol, 1 eq) and prop-2-enoyl chloride (1.03 mg, 1 1.34 μmol, 9.21e-1 μL, 0.3 eq) in DCM (1 mL) was added TEA (1 1 .47 mg, 1 13.38 μmol, 15.78 μL, 3 eq) .The mixture was stirred at 0°C for 1 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC TFA condition column: Phenomenex luna C18 100 x 40mm x 5 um; mobile phase: [H₂O (0.1 % TFA) - ACN]; gradient: 20%-50% B over 8.0 min to give 5-[2-(dimethylamino)ethoxy]-16-(5-methyl-4-prop-2-enoyl-2,3- dihydroquinoxalin-1-yl)-8-oxa-2,14,20,21-tetrazatetracyclo[12.6.2.1^3,7.0^18,22]tricosa- 1 (20),3,5,7(23),16,18,21-heptaen-15-one (3.19 mg, 4.36 μmol, 11.55% yield, 99.023% purity, TFA) as a yellow solid. Procedure for Preparation of Compound 009

Scheme 15

To a solution of tert-butyl 8-methyl-4-(2-methylsulfanyl-7-oxo-8H-pyrido[2,3- d]pyrimidin-6-yl)-2,3-dihydroquinoxaline-1-carboxylate (1.5 g, 3.4 mmol, 1 eq) in acetone (5 mL) was added2-(2-bromoethoxy)ethanol (1.7 g, 10.2 mmol, 3 eq) and Nal (102.3 mg, 682.6 μmol, 0.2 eq), Cs₂CO₃ (3.3 g, 10.2 mmol, 3 eq). The mixture was stirred at 60°C for 12 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with H₂O (20 mL) and extracted with Ethyl acetate (20 mL x 3). The combined organic layers were washed with brine (15 mL x 1 ), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (20 g Silica Flash Column, Eluent of 0 ~ 40% Ethyl acetate/Petroleum ether gradient @ 120 mL/min) to give benzyl 4-[7-[[1-[2-(tert-butoxycarbonylamino)ethyl]pyrazol-4-yl]amino]-1-(3- hydroxypropyl)-2-oxo-1 ,6-naphthyridin-3-yl]-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate as a yellow solid.

To a solution of tert-butyl 4-[8-[2-(2-hydroxyethoxy)ethyl]-2-methylsulfanyl-7-oxo- pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (950 mg, 1.8 mmol, 1 eq) in DCM (15 mL) was added m-CPBA (548.3 mg, 2.7 mmol, 85% purity, 1.5 eq) at 0°C. The mixture was stirred at 20°C for 12 hr. LC-MS showed desired compound was detected.

The residue was diluted with Na₂SO₃ (30 mL) and extracted with Dichloromethane (30 mL x 3). The combined organic layers were washed with NaHCO₃(10 mL x 1 ), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give tert-butyl 4-[8-[2-(2- hydroxyethoxy)ethyl]-2-methylsulfonyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3- dihydroquinoxaline-1 -carboxylate (1 g, crude) as a yellow solid.

To a solution of 2-fluoropyridin-4-amine (200.3 mg, 1.8 mmol, 2 eq) in THF (10 mL) was added LiHMDS (1 M, 3.6 mL, 4 eq) under N₂ atmosphere, the mixture was stirred at - 78°C for 15 min under N₂ atmosphere. Then added tert-butyl 4-[8-[2-(2- hydroxyethoxy)ethyl]-2-methylsulfonyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3- dihydroquinoxaline-1-carboxylate (500 mg, 893.4 μmol, 1 eq) in THF (3 mL) to the mixture, the mixture was stirred at 25°C for 12 hr under N₂ atmosphere. LC-MS showed desired compound was detected. The reaction mixture was quenched by addition NH₄CI (15 mL) and then extracted with ethyl acetate (20 mL x 3). The organic phase was separated, dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (12 g Silica Flash Column, Eluent of 0-80% Ethyl acetate/Petroleum ether gradient @ 80 mL/min) to give tert-butyl 4-[2-[(2- fluoro-4-pyridyl)amino]-8-[2-(2-hydroxyethoxy)ethyl]-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8- methyl-2,3-dihydroquinoxaline-1 -carboxylate (370 mg, 625.4 μmol, 70.0% yield) as a yellow solid.

To a solution of tert-butyl 4-[2-[(2-fluoro-4-pyridyl)amino]-8-[2-(2- hydroxyethoxy)ethyl]-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1- carboxylate (170 mg, 287.3 μmol, 1 eq), in DMF (17 mL) was added t-BuOK (193.5 mg, 1.7 mmol, 6 eq) under N₂ atmosphere. The mixture was stirred at 60°C for 12 hr. Then the mixture was stirred at 70°C for 2 hr under N₂ atmosphere. LC-MS showed desired compound was detected. The reaction mixture was partitioned between NH₄CL (100 mL) and ethyl acetate (100 mL x 3). The organic phase was separated, dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiO₂, Dichloromethane: Methanol = 10:1 ) to give tert-butyl 8-methyl-4- (15-oxo-8,11-dioxa-2,6,14,20,21-pentazatetracyclo[12.6.2.13,7.018,22]tricosa- 1 (20),3,5,7(23),16,18,21-heptaen-16-yl)-2,3-dihydroquinoxaline-1-carboxylate (200mg ) as a brown solid.

To a solution of tert-butyl 8-methyl-4-(15-oxo-8,11-dioxa-2,6,14,20,21- pentazatetracyclo[12.6.2.13,7.018,22]tricosa-1(20),3,5,7(23),16,18,21-heptaen-16-yl)-2,3- dihydroquinoxaline-1-carboxylate (180 mg, 314.9 μmol, 1 eq) in EtOAc (2 mL) was added HCI/EtOAc (2 mL,4M) at 0°C. The mixture was stirred at 0 °C for 2 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to give 16-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-8,11-dioxa-2,6,14,20,21- pentazatetracyclo[12.6.2.13,7.018,22]tricosa-1(20),3,5,7(23),16,18,21-heptaen-15-one (180 mg, crude) as a yellow solid.

To a solution of 16-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-8,11-dioxa- 2,6,14,20,21-pentazatetracyclo[12.6.2.13,7.018,22]tricosa-1 (20),3,5,7(23),16,18,21-heptaen- 15-one (180 mg, 381.7 μmol, 1 eq) in THF (2 mL) and H₂O (2 mL) was added NaHCO₃ (96.2 mg, 1 .2 mmol, 44.6 μL, 3 eq) and prop-2-enoyl chloride (34.5 mg, 381 .8 μmol, 31.0 μL, 1 eq) at 0°C. The mixture was stirred at 0°C for 30 min. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (TFA condition column: Phenomenex luna C18 100 x 40mm x 3 um; mobile phase: [H₂O(0.1% TFA) - ACN]; gradient: 20% - 50% B over 8.0 min). The residue was purified by prep-HPLC (NH₄HCO₃ condition column: Waters Xbridge BEH C18 100 x 30 mm x 10 um; mobile phase: [H₂O(10mM NH₄HCO₃)-ACN]; gradient: 25% - 55% B over 8.0 min) to give 16-(5-methyl-4-prop-2-enoyl-2,3-dihydroquinoxalin-1-yl)-8,11-dioxa-2,6,14,20,21- pentazatetracyclo[12.6.2.13,7.018,22]tricosa-1(20),3,5,7(23),16,18,21-heptaen-15-one (17.67 mg, 33.42 μmol, 8.75% yield, 99.40% purity) as a yellow solid.

Procedure for Preparation of Compound 010

Scheme 16

To a solution of benzyl 4-[8-[2-(2-hydroxyethoxy)ethyl]-2-methylsulfinyl-7-oxo- pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (200 mg, 346.2 μmol, 1 eq) in dioxane (2 mL) was added tert-butyl N-(3-aminophenyl)carbamate (72.1 mg, 346.2 μmol, 1 eq) and TFA (59.2 mg, 519.4 μmol, 38.6 μL, 1 .5 eq). The mixture was stirred at 80°C for 12 hr. LC-MS showed desired compound was detected. The residue was diluted with H₂O (60 mL) and extracted with Ethyl acetate (60 mL x 3), filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (12 g Silica Flash Column, Eluent of 0-100% Ethyl acetate/Petroleum ether gradient @ 120 mL/min) to give benzyl 4-[2-[3-(tert- butoxycarbonylamino)anilino]-8-[2-(2-hydroxyethoxy)ethyl]-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]- 8-methyl-2,3-dihydroquinoxaline-1-carboxylate (940 mg, 1.3 mmol, 94.0% yield) as a yellow solid.

To a solution of benzyl 4-[2-[3-(tert-butoxycarbonylamino)anilino]-8-[2-(2- hydroxyethoxy)ethyl]-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1- carboxylate (940.0 mg, 1.3 mmol, 1 eq) in DCM (10 mL) was added TEA (395.3 mg, 3.9 mmol, 543.8 μL, 3 eq), DMAP (15.9 mg, 130.2 μmol, 0.1 eq) and TosCI (496.6 mg, 2.6 mmol, 2 eq) at 0°C. The mixture was stirred at 20°C for 12 hr. LC-MS showed 44% of Reactant 1 remained. The mixture was added TosCI (124.1 mg, 651.1 μmol, 0.5 eq) and DMAP (31.8 mg, 260.5 μmol, 0.2 eq). The mixture was stirred at 20°C for 12 hr. LC-MS showed desired compound was detected. The residue was diluted with NH₄CI (50 mL) and extracted with Dichloromethane (50 mL x 3). The combined organic layers were washed with NaHCO₃ (20 mL x 2), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (4 g Silica Flash Column, Eluent of 0-100% Ethyl acetate/Petroleum ether gradient @ 120 mL/min) to give benzyl 4-[2-[3-(tert-butoxycarbonylamino)anilino]-7-oxo-8-[2-[2-(p- tolylsulfonyloxy)ethoxy]ethyl]pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1- carboxylate (500 mg, 570.8 μmol, 43.8% yield) as a yellow solid.

To a solution of benzyl 4-[2-[3-(tert-butoxycarbonylamino)anilino]-7-oxo-8-[2-[2-(p- tolylsulfonyloxy)ethoxy]ethyl]pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1 - carboxylate (500 mg, 570.8 μmol, 1 eq) in THF (40 mL) was added Nal (8.6 mg, 57.1 μmol, 0.1 eq) and NaH (27.4 mg, 684.9 μmol, 60% purity, 1 .2 eq) at 0°C. The mixture was stirred at 80°C for 12 hr under N₂ atmosphere. LC-MS showed desired compound was detected. The residue was diluted with NH₄CI (60 mL) and extracted with Ethyl acetate (60 mL ×3), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (TFA condition column: Phenomenex luna C18 100 x 40mm x 3 um; mobile phase: [H₂O(0.1% TFA)-ACN]; gradient: 65% - 85% B over 8.0 min) to give tert-butyl 16-(4-benzyloxycarbonyl-5-methyl-2,3-dihydroquinoxalin-1-yl)-15-oxo-11-oxa- 2,8,14,20,21-pentazatetracyclo[12.6.2.13,7.018,22]tricosa-1 (20),3,5,7(23),16,18,21- heptaene-8-carboxylate (50 mg, 71.0 μmol, 12.5% yield) as a yellow solid.

To a solution of Pd(OH)₂ (100 mg, 20% purity) in MeOH (5 mL) was added tert-butyl 16-(4-benzyloxycarbonyl-5-methyl-2,3-dihydroquinoxalin-1-yl)-15-oxo- 11-oxa-2,8,14,20,21- pentazatetracyclo[12.6.2.13,7.018,22]tricosa-1(20),3,5,7(23),16,18,21-heptaene-8- carboxylate (70 mg, 99.5 μmol, 1 eq) and TEA (30.2 mg, 298.4 μmol, 41.53 μL, 3 eq) under N₂ atmosphere . The suspension was degassed and purged with H₂ for 3 times. The mixture was stirred under H₂ (15Psi) at 20°C for 1 hr. LC-MS showed desired compound was detected. The reaction mixture was filtered and concentrated under reduced pressure to give tert-butyl 16-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-15-oxo-11-oxa-2,8,14,20,21- pentazatetracyclo[12.6.2.13,7.018,22]tricosa-1(20),3,5,7(23),16,18,21-heptaene-8- carboxylate (45 mg, crude) as a yellow solid.

To a solution of tert-butyl 16-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-15-oxo-11- oxa-2,8,14,20,21-pentazatetracyclo[12.6.2.13,7.018,22]tricosa-1(20),3,5,7(23),16,18,21- heptaene-8-carboxylate (45 mg, 79.0 μmol, 1 eq) in DCM (0.5 mL) was added TEA (24.0 mg, 237.0 μmol, 33.0 μL, 3 eq) and prop-2-enoyl chloride (7.2 mg, 79.0 μmol, 6.42 μL, 1 eq) at 0°C. The mixture was stirred at 20°C for 1 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (TFA condition column: Phenomenex luna C18 100 x 40 mm x 3 um; mobile phase: [H₂O(0.1 % TFA)-ACN]; gradient: 30% - 75% B over 8.0 min) to tert-butyl 16-(5-methyl-4-prop-2-enoyl-2,3-dihydroquinoxalin-1-yl)-15-oxo- 11-oxa-2,8, 14,20,21 pentazatetracyclo[12.6.2.13,7.018,22]tricosa-1(20),3,5,7(23),16,18,21- heptaene-8-carboxylate (40 mg, 54.2 μmol, 68.6% yield, TFA) as a yellow solid.

To a solution of tert-butyl 16-(5-methyl-4-prop-2-enoyl-2,3-dihydroquinoxalin-1-yl)-15- oxo-11-oxa-2,8,14,20,21-pentazatetracyclo[12.6.2.13,7.018,22]tricosa- 1 (20),3,5,7(23),16,18,21-heptaene-8-carboxylate (40 mg, 64.1 μmol, 1 eq) in DCM (1 mL) was added TFA (460.5 mg, 4.0 mmol, 0.3 mL, 63.0 eq). The mixture was stirred at 20°C for 1 .5 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep- HPLC (TFA condition column: Phenomenex Luna C18 75 x 30mm x 3um; mobile phase: [H₂O(0.1% TFA)-ACN]; gradient: 20% - 50% B over 8.0 min) to give 6-(5-methyl-4-prop-2- enoyl-2,3-dihydroquinoxalin-1 -yl)-11 -oxa-2,8, 14,20,21 - pentazatetracyclo[12.6.2.13,7.018,22]tricosa-1(20),3,5,7(23),16,18,21-heptaen-15-one (6.9 mg, 13.18 μmol, 20.55% yield, 100% purity) as a yellow solid.

Procedure for Preparation of Compound 016

Scheme 17

To a solution of benzyl 8-methyl-4-(2-methylsulfanyl-7-oxo-8H-pyrido[2,3-d]pyrimidin- 6-yl)-2,3-dihydroquinoxaline-1-carboxylate (2 g, 4.2 mmol, 1 eq) in acetone (70 mL) was added Nal (126.6 mg, 844.6 μmol, 0.2 eq), 2-(2-bromoethoxy)ethanol (1.2 g, 7.6 mmol, 1.8 eq) and Cs₂CO₃ (4.1 g, 12.6 mmol, 3 eq). The mixture was stirred at 60°C for 12 hr. TLC indicated benzyl 8-methyl-4-(2-methylsulfanyl-7-oxo-8H-pyrido[2,3-d]pyrimidin-6-yl)-2,3- dihydroquinoxaline-1 -carboxylate was consumed completely. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with H₂O (40 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with brine (20 mL x 2), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (40 g Silica Flash Column, Eluent of 0-100% Ethyl acetate/Petroleum ether gradient @ 100 mL/min) to give benzyl 4-[8-[2-(2-hydroxyethoxy)ethyl]-2-methylsulfanyl-7-oxo-pyrido[2,3- d]pyrimidin-6-yl]-8-methyl2,3-dihydroquinoxaline-1-carboxylate (1 g, 1.7 mmol, 40.4% yield, 96% purity) as a yellow solid.

To a solution of benzyl 4-[8-[2-(2-hydroxyethoxy)ethyl]-2-methylsulfanyl-7-oxo pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (1.7 g, 3.1 mmol, 1 eq) in DCM (30 mL) was added m-CPBA (965.1 mg, 4.7 mmol, 85% purity, 1.5 eq) at 0°C. The resulting mixture was stirred at 25°C for 2 hr. LC-MS showed desired compound was detected.

The reaction mixture was quenched by addition Na₂SO₃ (20 mL), extracted with dichloromethane (20 mL x 3). The combined organic layers were washed with NaHCO₃ (20 mL x 2), and dried over Na₂SO₄, filtered and concentrated under reduced pressure to give benzyl 4-[8-[2-(2-hydroxyethoxy)ethyl]-2-methylsulfinyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8- methyl2,3-dihydroquinoxaline-1 -carboxylate (1.7 g, crude) as a red solid.

To a solution of benzyl 4-[8-[2-(2-hydroxyethoxy)ethyl]-2-methylsulfinyl-7-oxo- pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (300 mg, 519.3 μmol, 1 eq) in dioxane (3 mL) was added TFA (88.8 mg, 779.0 μmol, 57.8 μL, 1 .5 eq) and tert-butyl 6-aminoindoline-1 -carboxylate (146.0 mg, 623.2 μmol, 1.2 eq). The mixture was stirred at 80°C for 12 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by flash silica gel chromatography (20 g Silica Flash Column, Eluent of 0-45% Ethyl acetate/Petroleum ether gradient @ 100 mL/min) to give benzyl 4-[2-[(1-tert- butoxycarbonylindolin-6-yl)amino]-8-[2-(2-hydroxyethoxy)ethyl]-7-oxo-pyrido[2,3- d]pyrimidin- 6-yl]-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (616.9 mg, 775.4 μmol, 49.7% yield, 94% purity) as yellow oil.

To a solution of benzyl 4-[2-[(1-tert-butoxycarbonylindolin-6-yl)amino]-8-[2-(2- hydroxyethoxy)ethyl]-7-oxopyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1- carboxylate (616.9 mg, 824.9 μmol, 1 eq) in DCM (10 mL) was added TosCI (172.9 mg, 907.4 μmol, 1.1 eq), TEA (183.6 mg, 1.8 mmol, 252.6 μL, 2.2 eq) and DMAP (10.0 mg, 82.4 μmol, 0.1 eq) at 0°C. The mixture was stirred at 25°C for 12 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by flash silica gel chromatography (20 g Silica Flash Column, Eluent of 0-50% Ethyl acetate/Petroleum ether gradient @ 100 mL/min) to give benzyl 4-[2-[(1-tert-butoxycarbonylindolin-6-yl)amino]-7-oxo-8-[2-[2- (ptolylsulfonyloxy)ethoxy]ethyl]pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline- 1-carboxylate (539 mg, 424.2 μmol, 51.4% yield, 71 % purity) as a yellow solid.

A solution of benzyl 4-[2-[(1-tert-butoxycarbonylindolin-6-yl)amino]-7-oxo-8-[2- [2-(p- tolylsulfonyloxy)ethoxy]ethyl]pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1- carboxylate (539 mg, 597.5 μmol, 1 eq) in TFA (1 mL) and DCM (4 mL) was stirred at 25°C for 1 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to give benzyl 4-[2-(indolin-6-ylamino)-7-oxo-8-[2-[2- (p-tolylsulfonyloxy)ethoxy]ethyl]pyrido[2,3-d]pyrimidin6-yl]-8-methyl-2,3dihydroquinoxaline-1- carboxylate (547.3 mg, crude, TFA) as a yellow solid.

To a solution of benzyl 4-[2-(indolin-6-ylamino)-7-oxo-8-[2-[2-(p- tolylsulfonyloxy)ethoxy]ethyl]pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1- carboxylate (547.3 mg, 597.5 μmol, 1 eq, TFA) in DMF (6 mL) was added K₂CO₃ (247.7 mg, 1 .7 mmol, 3 eq) and TEA (532.4 mg, 5.2 mmol, 732.3 μL, 8.8 eq) to adjust pH to 7-8. The mixture was stirred at 60°C for 12 h. LC-MS showed desired compound was detected. The residue was diluted with H₂O (50 mL) and extracted with ethyl acetate (50 mL x 3). The combined organic layers were washed with brine (40 mL x 3), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (20 g Silica Flash Column, Eluent of 0-100% Ethyl acetate/Petroleum ether gradient @ 100 mL/min) to give benzyl 8-methyl-4-(16-oxo-12-oxa- 2,9,15,21 ,22- pentazapentacyclo[13.6.2.23,6.05,9.019,23]pentacosa1 (21 ),3(25),4,6(24),17,19,22-heptaen- 17-yl)-2,3-dihydroquinoxaline-1 -carboxylate (204 mg, 304.5 μmol, 50.9% yield, 94% purity) as a yellow solid.

To a solution of benzyl 8-methyl-4-(16-oxo-12-oxa-2,9,15,21 ,22- pentazapentacyclo[13.6.2.23,6.05,9.019,23]pentacosa1 (21 ),3(25),4,6(24),17,19,22-heptaen- 17-yl)-2,3-dihydroquinoxaline-1 -carboxylate (204 mg, 323.9 μmol, 1 eq) in EtOAc (5 mL) MeOH (5 mL) was added Pd(OH)₂ (100 mg, 142.4 μmol, 20% purity, 0.4 eq) under N₂ atmosphere. The suspension was added TEA to adjust pH to 8. The suspension was degassed and purged with H₂ for 3 times. The mixture was stirred under H₂ (15 Psi) at 20°C for 12 hr. LC-MS showed desired compound was detected. The reaction mixture was filtered and concentrated under reduced pressure to give 17-(5-methyl-3,4-dihydro-2H- quinoxalin-1-yl)-12-oxa-2,9, 15,21 ,22-pentazapentacyclo[13.6.2.23,6.05,9.019,23]pentacosa- 1 (21 ),3(25),4,6(24),17,19,22-heptaen-16-one (59 mg, crude) as yellow oil.

To a solution of 17-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-12-oxa-2,9,15,21 ,22- pentazapentacyclo[13.6.2.23,6.05,9.019,23]pentacosa-1 (21),3(25),4,6(24),17,19,22- heptaen-16-one (29.5 mg, 59.5 μmol, 1 eq) in THF (1.6 mL) and H₂O (0.4 mL) was added NaHCO₃ (15.0 mg, 178.5 μmol, 6.9 μL, 3 eq). The mixture was added dropwise prop-2- enoyl chloride (5.3 mg, 59.5 μmol, 4.8 μL, 1 eq) in THF (0.1 mL) at 0°C and stirred at 0°C for 0.5 hr. LC-MS showed 17-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-12-oxa-2,9,15,21 ,22- pentazapentacyclo[13.6.2.23,6.05,9.019,23]pentacosa-1 (21),3(25),4,6(24),17,19,22- heptaen-16-one remained. The mixture was added dropwise prop-2-enoyl chloride (2.6 mg, 29.7 μmol, 2.4 μL, 0.5 eq) in THF (0.1 mL) at 0°C and stirred at 0°C for 0.5 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (TFA condition; column: Phenomenex luna C18 100x40mmx3 um; mobile phase: [H₂O(0.1% TFA)-ACN];gradient:35%-65% B over 8.0 min) to give 17-(5-methyl-4-prop-2-enoyl-2,3- dihydroquinoxalin-1-yl)-12-oxa-2,9,15,21 ,22- pentazapentacyclo[13.6.2.23,6.05,9.019,23]pentacosa-1 (21),3(25),4,6(24),17,19,22- heptaen-16-one (18.1 mg, 32.0 μmol, 26.8% yield, 97% purity) as a yellow solid.

Procedure for Preparation of Compound 035

Scheme 18

To a solution of tert-butyl N-(3-hydroxypropyl)-N-methyl-carbamate (1 g, 5.3 mmol, 1 eq) in DCM (10 mL) was added DMAP (64.6 mg, 528.4 μmol, 0.1 eq), TEA (1.2 g, 11.6 mmol, 1.6 mL, 2.2 eq) and TsCI (1.1 g, 5.8 mmol, 1.1 eq) at 0°C. The mixture was stirred at 20°C for 12 hr. LC-MS showed desired compound was detected. The reaction mixture was quenched by addition NH₄CI (30 mL), and then diluted with dichloromethane (30 mL x 3) and extracted with NaHCO₃ (30 mL). The combined organic layers were washed with aqueous NaCI (15 mL x 1 ), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give 3-[tert-butoxycarbonyl(methyl)amino]propyl 4-methylbenzenesulfonate (1.8 g, crude) as brown oil.

To a solution of benzyl 8-methyl-4-(2-methylsulfanyl-7-oxo-8H-pyrido[2,3-d]pyrimidin- 6-yl)-2,3-dihydroquinoxaline-1-carboxylate (2.5 g, 5.3 mmol, 1 eq) in acetone (40 mL) was added 3-[tert-butoxycarbonyl(methyl)amino]propyl 4-methylbenzenesulfonate (2.2 g, 6.3 mmol, 1 .2 eq), Nal (158.3 mg, 1.1 mmol, 0.2 eq) and Cs₂CO₃ (5.2 g, 15.8 mmol, 3 eq). The mixture was stirred at 60°C for 12 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with H₂O (150 mL) and extracted with ethyl acetate (150 mL x 3). The combined organic layers were washed with brine (130 mL x 1 ), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (20 g Silica Flash Column, Eluent of 0-26% Ethyl acetate/Petroleum ether gradient @ 120 mL/min) to give benzyl 4-[8-[3-[tert- butoxycarbonyl(methyl)amino]propyl]-2-methylsulfanyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8- methyl-2,3-dihydroquinoxaline-1-carboxylate (1.5 g, 2.3 mmol, 42.6% yield) as a yellow solid.

To a solution of benzyl 4-[8-[3-[tert-butoxycarbonyl(methyl)amino]propyl]-2- methylsulfanyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1- carboxylate (1.3 g, 1 .9 mmol, 1 eq) in DCM (5 mL) was added m-CPBA (708.4 mg, 3.5 mmol, 85% purity, 1 .8 eq) at 0°C. The mixture was stirred at 0°C for 2 hr. LC-MS showed desired compound was detected. The residue was diluted with Na₂SO₃ (150 mL) and extracted with dichloromethane (150 mL x 3). The combined organic layers were washed with NaHCO₃ (130 mL x 3), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give benzyl 4-[8-[3-[tert-butoxycarbonyl(methyl)amino]propyl]-2-methylsulfonyl-7- oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (1.4 g, crude) as a yellow solid.

To a solution of benzyl 4-[8-[3-[tert-butoxycarbonyl(methyl)amino]propyl]-2- methylsulfonyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1- carboxylate (200 mg, 295.5 μmol, 1 eq) in dioxane (2 mL) was added TFA (50.5 mg, 443.3 μmol, 32.9 μL, 1.5 eq) and methyl 3-aminobenzoate (53.6 mg, 354.6 μmol, 1.2 eq). The mixture was stirred at 80°C for 12 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with NaHCO₃ (50 mL) and extracted with ethyl acetate (50 mL x 3). The combined organic layers were washed with brine (15 mL x 1 ), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (12 g Silica Flash Column, Eluent of 0-75% Ethyl acetate/Petroleum ether gradient @ 120 mL/min) to give benzyl 4-[8-[3-[tert- butoxycarbonyl(methyl)amino]propyl]-2-(3-methoxycarbonylanilino)-7-oxo-pyrido[2,3- d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (420 mg, 561.6 μmol, 63.4% yield) as a yellow solid.

A mixture of benzyl 4-[8-[3-[tert-butoxycarbonyl(methyl)amino]propyl]-2-(3- methoxycarbonylanilino)-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline- 1-carboxylate (300 mg, 401.2 μmol, 1 eq) in THF (6 mL) and H₂O (3 mL) was added LiOH·H₂O (84.2 mg, 2.0 mmol, 5 eq), the mixture was stirred at 80°C for 2 hr. LC-MS showed desired compound was detected. The reaction mixture was acidified to pH=4 with 4 M HCI, and then diluted with H₂O (30 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with aqueous NaCI (25 mL x 1), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give 3-[[6-(4-benzyloxycarbonyl-5- methyl-2,3-dihydroquinoxalin-1-yl)-8-[3-[tert-butoxycarbonyl(methyl)amino]propyl]-7-oxo- pyrido[2,3-d]pyrimidin-2-yl]amino]benzoic acid (290 mg, crude) as a yellow solid.

A mixture of 3-[[6-(4-benzyloxycarbonyl-5-methyl-2,3-dihydroquinoxalin-1-yl)-8-[3- [tert-butoxycarbonyl(methyl)amino]propyl]-7-oxo-pyrido[2,3-d]pyrimidin-2-yl]amino]benzoic acid (290 mg, 395.2 μmol, 1 eq) in HCI/EtOAc (5 mL, 4 M) was stirred at 20°C for 1 hr. LC- MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent to give 3-[[6-(4-benzyloxycarbonyl-5-methyl-2,3- dihydroquinoxalin-1-yl)-8-[3-(methylamino)propyl]-7-oxo-pyrido[2,3-d]pyrimidin-2- yl]amino]benzoic acid (300 mg, crude, HCI) as a yellow solid.

To a solution of 3-[[6-(4-benzyloxycarbonyl-5-methyl-2,3-dihydroquinoxalin-1-yl)-8-[3- (methylamino)propyl]-7-oxo-pyrido[2,3-d]pyrimidin-2-yl]amino]benzoic acid (200 mg, 315.6 μmol, 1 eq) in Py (30 mL) was added EDCI (90.8 mg, 473.4 μmol, 1.5 eq). The mixture was stirred at 50°C for 12 hr. LC-MS showed desired compound was detected.

The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with NH₄CI (30 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (20 mL x 1), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiO₂, DCM: MeOH = 20:1 ). The residue was purified by prep-HPLC(TFA condition, column: Phenomenex luna C18 100 x 40 mm x 3 um; mobile phase: [H₂O (0.1% TFA)- ACN]; gradient: 35 % - 65 % B over 8.0 min) to give benzyl 8-methyl-4-(9-methyl-8,14- dioxo-2,9,13,19,20-pentazatetracyclo[11.6.2.13,7.017,21]docosa-1 (19),3,5,7(22),15,17,20- heptaen-15-yl)-2,3-dihydroquinoxaline-1 -carboxylate (60 mg) as a yellow solid.

To a solution of Pd(OH)₂/C (20 mg, 20% purity) in MeOH (2 mL) was added benzyl 8- methyl-4-(9-methyl-8, 14-dioxo-2,9, 13,19,20-pentazatetracyclo[11 .6.2.13,7.017,21 ]docosa- 1 (19),3,5,7(22),15,17,20-heptaen-15-yl)-2,3-dihydroquinoxaline-1-carboxylate (40 mg, 65.0 μmol, 1 eq) under N₂ atmosphere. The suspension was degassed and purged with H₂ for 3 times. The mixture was stirred under H₂ (15 psi) at 20°C for 1 hr. LC-MS showed desired compound was detected. The reaction mixture was filtered and concentrated under reduced pressure to give 9-methyl-15-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-2,9,13,19,20- pentazatetracyclo[11 ,6.2.13,7.017,21]docosa-1 (19),3,5,7(22),15,17,20-heptaene-8,14-dione (40 mg, crude) as a yellow solid.

To a solution of 9-methyl-15-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-2,9, 13, 19,20- pentazatetracyclo[11 ,6.2.13,7.017,21]docosa-1 (19),3,5,7(22),15,17,20-heptaene-8,14-dione (40 mg, 83.1 μmol, 1 eq) in DCM (1 mL) was added TEA (25.2 mg, 249.2 μmol, 34.7 μL, 3 eq) and prop-2-enoyl chloride (7.5 mg, 83.1 μmol, 6.8 μL, 1 eq) at 0°C. The mixture was stirred at 0°C for 1 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (TFA condition, column: Phenomenex Luna C 18 100 x 40 mm x 5 um; mobile phase: [H₂O(0.1% TFA)- ACN]; gradient: 35 % - 65 % B over 8.0 min) to give 9- methyl-15-(5-methyl-4-prop-2-enoyl-2,3-dihydroquinoxalin-1-yl)-2,9, 13, 19,20- pentazatetracyclo[11 ,6.2.13,7.017,21]docosa-1 (19),3,5,7(22),15,17,20-heptaene-8,14-dione (12.66 mg, 23.6 μmol, 28.5% yield, 100% purity) as a yellow solid.

VT ¹H NMR (400 MHz, DMSO-d₆) δ = 10.01 (s, 1 H), 8.75 (s, 1 H), 8.09 (s, 1 H), 7.86 (s, 1 H), 7.45 - 7.37 (m, 1H), 7.22 (br d, J = 8.1 Hz, 1 H), 7.16 (d, J = 7.6 Hz, 1 H), 6.90 (br t, J = 7.9 Hz, 1 H), 6.64 (br d, J = 7.3 Hz, 1H), 6.36 (br d, J = 8.3 Hz, 1H), 6.30 - 6.21 (m, 1 H), 5.70 (br dd, J = 4.1 , 6.3 Hz, 1H), 4.97 - 4.76 (m, 1 H), 4.03 - 3.86 (m, 2H), 3.76 - 3.62 (m, 1 H), 3.58 - 3.33 (m, 3H), 3.15 - 2.98 (m, 1 H), 2.95 (s, 3H), 2.24 - 2.08 (m, 5H). Procedure for Preparation of Compound 036

Scheme 19

To a solution of 4-nitro-1 H-pyrazole (4.5 g, 39.8 mmol, 1 eq) in ACN (80 mL) was added 2-bromo-1 ,1 -dimethoxy-ethane (10.1 g, 59.7 mmol, 7.0 mL, 1.5 eq) and Cs₂CO₃ (19.5 g, 59.7 mmol, 1.5 eq). The mixture was stirred at 90°C for 12 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with H₂O (100 mL) and extracted with Ethyl acetate (100 mL x 3), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (20 g Silica Flash Column, Eluent of 0-100% Ethyl acetate/Petroleum ether gradient @ 120 mL/min) to give 1-(2,2-dimethoxyethyl)-4-nitro-pyrazole (7 g, 34.8 mmol, 87.4% yield) as a white solid.

To a solution of PtO₂ (500 mg) in EtOAc (20 mL) and EtOH (60 mL) was added 1- (2,2-dimethoxyethyl)-4-nitro-pyrazole (3 g, 14.9 mmol, 1 eq) under N₂ atmosphere. The suspension was degassed and purged with H₂ for 3 times. The mixture was stirred under H₂ (15Psi) at 20°C for 12 hr. LC-MS showed desired compound was detected. The residue was filtered and concentrated under reduced pressure to give a residue to give 1 -(2,2- dimethoxyethyl)pyrazol-4-amine (2.8 g, crude) as a purple oil.

To a solution of benzyl 4-[8-[2-[tert-butoxycarbonyl(methyl)amino]ethyl]-2- methylsulfanyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1- carboxylate (1 g, 1 .6 mmol, 1 eq) in DCM (10 mL) was added and m-CPBA (482.8 mg, 2.4 mmol, 85% purity, 1 .5 eq) at 0°C. The mixture was stirred at 20°C for 2 hr. LC-MS showed desired compound was detected. The residue was diluted with Na₂SO₃ (50 mL) and extracted with Dichloromethane (50 mL x 3). The combined organic layers were washed with NaHCO₃ (20 mL x 2), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue to give benzyl 4-[8-[2-[tert-butoxycarbonyl(methyl)amino]ethyl]-2- methylsulfonyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1- carboxylate (1.15 g, crude) as a yellow solid.

To a solution of benzyl 4-[8-[2-[tert-butoxycarbonyl(methyl)amino]ethyl]-2- methylsulfonyl-7-oxo-pyrido[2 , 3- d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (200 mg, 301.8 μmol, 1 eq) in dioxane (2 mL) was added TFA (51.6 mg, 452.7 μmol, 33.6 μL, 1.5 eq) and 1-(2,2- dimethoxyethyl)pyrazol-4-amine (77.5 mg, 452.7 μmol, 1.5 eq). The mixture was stirred at 80°C for 12 hr. LC-MS showed desired compound was detected. The residue was diluted with H₂O 20 mL and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with brine (15 mL x 1 ), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (12 g Silica Flash Column, Eluent of 0-55% Ethyl acetate/Petroleum ether gradient @ 120 mL/min) to give benzyl 4-[8-[2-[tert-butoxycarbonyl(methyl)amino]ethyl]-2- [[1-(2,2-dimethoxyethyl)pyrazol-4-yl]amino]- 7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (469 mg, 622.1 μmol, 68.7% yield) as a yellow solid.

A mixture of benzyl 4-[8-[2-[tert-butoxycarbonyl(methyl)amino]ethyl]-2-[[1-(2,2- dimethoxyethyl)pyrazol-4-yl]amino]-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3- dihydroquinoxaline-1-carboxylate (469 mg, 622.1 μmol, 1 eq) in HCI (4 mL) and EtOAc (5 mL) was stirred at 20°C for 30 min. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (HCI condition column: Phenomenex Luna C18 75 x30 mm x 3 um; mobile phase: [H₂O(0.04% HCI)-ACN]; gradient:20% - 50% B over 8.0 min) to give benzyl 8-methyl-4-[8-[2-(methylamino)ethyl]-7-oxo-2-[[1-(2-oxoethyl)pyrazol-4- yl]amino]pyrido[2,3-d]pyrimidin-6-yl]-2,3-dihydroquinoxaline-1-carboxylate (180 mg, 296.2 μmol, 47.6% yield) as a yellow solid.

To a solution of benzyl 8-methyl-4-[8-[2-(methylamino)ethyl]-7-oxo-2-[[1-(2- oxoethyl)pyrazol-4-yl]amino]pyrido[2,3-d]pyrimidin-6-yl]-2,3-dihydroquinoxaline-1 -carboxylate (50 mg, 82.3 μmol, 1 eq) in DCE (2.5 mL) and MeOH (5 mL) was added AcOH (14.8 mg, 246.9 μmol, 14.1 μL, 3 eq). The mixture was stirred at 20°C for 30 min. Then the mixture was added NaBH₃CN (10.3 mg, 164.6 μmol, 2 eq). The mixture was stirred at 20°C for 12 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep- HPLC (NH₄HCO₃ condition column: Waters Xbridge BEH C18 100 x 30 mm x 10 um; mobile phase: [H₂O(10mM NH₄HCO₃)-ACN]; gradient:65%-95% B over 8.0 min) to give benzyl 8- methyl-4-(9-methyl-13-oxo-2,5,6,9, 12, 18,19-heptazatetracyclo[10.6.2.13,6.016,20]henicosa- 1 (18), 3(21 ), 4, 14,16, 19-hexaen-14-yl)-2,3-dihydroquinoxaline-1 -carboxylate (70 mg) as a white solid.

To a solution of Pd(OH)₂ (100 mg, 20% purity) in THF (5 mL) was added benzyl 8- methyl-4-(9-methyl-13-oxo-2,5,6,9, 12, 18,19-heptazatetracyclo[10.6.2.13,6.016,20]henicosa- 1 (18), 3(21 ), 4, 14,16, 19-hexaen-14-yl)-2,3-dihydroquinoxaline-1 -carboxylate (70 mg, 118.3 μmol, 1 eq) under N₂ atmosphere. The suspension was degassed and purged with H₂ for 3 times. The mixture was stirred under H₂ (15Psi) at 20°C for 1 hr. LC-MS showed desired compound was detected. The residue was filtered and concentrated under reduced pressure to give 9-methyl-14-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-2,5,6,9,12,18,19- heptazatetracyclo[10.6.2.13,6.016,20]henicosa-1(18),3(21 ),4,14,16,19-hexaen-13-one (70 mg, crude) as a yellow solid. To a solution of 9-methyl-14-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-

2,5,6,9,12,18,19-heptazatetracyclo[10.6.2.13,6.016,20]henicosa-1 (18),3(21 ),4,14,16,19- hexaen-13-one (50 mg, 109.3 μmol, 1 eq) in DCM (0.5 mL) was added TEA (33.2 mg, 327.9 μmol, 45.6 μL, 3 eq) and prop-2-enoyl chloride (2.5 mg, 27.3 μmol, 2.2 μL, 0.3 eq) at 0°C. The mixture was stirred at 0°C for 30 min. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (FA condition column: Phenomenex Luna C18 100 x 30 mm x 5um; mobile phase: [H₂O(0.2% FA)-ACN]; gradient: 10%-40% B over 8.0 min) to give 9-methyl-14-(5-methyl-4-prop-2-enoyl-2,3-dihydroquinoxalin-1 -yl)-2,5,6,9, 12, 18, 19- heptazatetracyclo[10.6.2.13,6.016,20]henicosa-1(18),3(21 ),4,14,16,19-hexaen-13-one (560.00 μg, 9.94^e-1 μmol, 9.09^e-1% yield, 98.95% purity, FA) as a yellow solid.

Procedure for Preparation of Compound 002

Scheme 20

To a solution of benzyl 4-[8-(1-tert-butoxycarbonyl-3-piperidyl)-2-methylsulfinyl-7-oxo- pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (2 g, 3.0 mmol, 1 eq) in Tol. (20 mL) was added DIEA (1.2 g, 8.9 mmol, 1.6 mL, 3 eq) and 4-aminobutan-1-ol (318.0 mg, 3.6 mmol, 331.2 μL, 1.2 eq). The mixture was stirred at 25°C for 1 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by flash silica gel chromatography (20 g Silica Flash Column, Eluent of 0-60% Ethyl acetate/Petroleum ether gradient @ 100 mL/min) to give benzyl 4-[8-(1-tert-butoxycarbonyl-3-piperidyl)-2-(4- hydroxybutylamino)-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1- carboxylate (1.1 g, 1.6 mmol, 54.5% yield) as a yellow solid.

To a solution of benzyl 4-[8-(1-tert-butoxycarbonyl-3-piperidyl)-2-(4- hydroxybutylamino)-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1- carboxylate (1.1 g, 1.5 mmol, 1 eq) in DCM (12 mL) was added TEA (778.6 mg, 7.7 mmol, 1.1 mL, 5 eq), DMAP (18.8 mg, 153.9 μmol, 0.1 eq) and TosCI (880.2 mg, 4.6 mmol, 3 eq). The mixture was stirred at 25°C for 1 hr. LC-MS showed desired compound was detected. The residue was diluted with NaHCO₃ (20 mL) and extracted with dichloromethane (30 mL x 3). The combined organic layers were washed with brine (20 mL x 1 ), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (12 g Silica Flash Column, Eluent of 0-42% Ethyl acetate/Petroleum ether gradient @ 120 mL/min) to give benzyl 4-[8-(1-tert-butoxycarbonyl- 3-piperidyl)-7-oxo-2-[4-(p-tolylsulfonyloxy)butylamino]pyrido[2,3-d]pyrimidin-6-yl]-8-methyl- 2, 3-dihydroquinoxaline-1 -carboxylate (830 mg, 974.2 μmol, 63.3% yield) as a yellow solid.

A mixture of benzyl 4-[8-(1-tert-butoxycarbonyl-3-piperidyl)-7-oxo-2-[4-(p- tolylsulfonyloxy)butylamino]pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1- carboxylate (830 mg, 974.2 μmol, 1 eq) in HCI/EtOAc (4 mL, 4 M) and EtOAc (4 mL) was stirred at 25°C for 1 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent to give benzyl 8- methyl-4-[7-oxo-8-(3-piperidyl)-2-[4-(p-tolylsulfonyloxy)butylamino]pyrido[2,3-d]pyrimidin-6- yl]-2,3-dihydroquinoxaline-1 -carboxylate (800 mg, crude, HCI) as a yellow solid.

To a solution of benzyl 8-methyl-4-[7-oxo-8-(3-piperidyl)-2-[4-(p- tolylsulfonyloxy)butylamino]pyrido[2,3-d]pyrimidin-6-yl]-2,3-dihydroquinoxaline-1 -carboxylate (800 mg, 1 .0 mmol, 1 eq, HCI) in DMF (8 mL) was added K₂CO₃ (420.8 mg, 3.0 mmol, 3 eq). The mixture was stirred at 100°C for 2 hr. LC-MS showed desired compound was detected. The residue was diluted with H₂O (20 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (20 mL x 2), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (TFA condition, column: Phenomenex luna C18 100 x 40 mm x 5 um; mobile phase: [H₂O(0.1% TFA)-ACN];gradient:27%-57% B over 8.0 min) to give benzyl 8-methyl-4- (18-oxo-1 ,6,11 ,13,20-pentazatetracyclo[10.6.2.12,6.015,19]henicosa-12,14,16,19-tetraen- 17-yl)-2,3-dihydroquinoxaline-1 -carboxylate (230 mg, 396.8 μmol, 39.1 % yield) as a yellow solid.

To a solution of Pd(OH)₂ (100 mg, 20% purity) in THF (10 mL) was added benzyl 8- methyl-4-( 18-oxo-1 ,6,11 ,13,20-pentazatetracyclo[10.6.2.12,6.015,19]henicosa- 12,14,16,19- tetraen-17-yl)-2,3-dihydroquinoxaline-1-carboxylate (100 mg, 172.5 μmol, 1 eq) and TEA (52.4 mg, 517.5 μmol, 72.0 μL, 3 eq) under N₂ atmosphere. The suspension was degassed and purged with H₂ for 3 times. The mixture was stirred under H₂ (15 Psi) at 25 °C for 0.5 hr. LC-MS showed desired compound was detected. The reaction mixture filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep- HPLC (TFA condition column: Phenomenex luna C18 100 x 40 mm x 5 um; mobile phase: [H₂O(0.1% TFA)-ACN]; gradient:8%-38% B over 8.0 min) to give 17-(5-methyl-3,4-dihydro- 2H-quinoxalin-1-yl)-1 ,6,11 ,13,20-pentazatetracyclo[10.6.2.12,6.015,19]henicosa- 12,14,16,19-tetraen-18-one (55 mg, 123.4 μmol, 71.6% yield) as a yellow solid.

To a solution of 17-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-1 ,6,11 ,13,20- pentazatetracyclo[10.6.2.12,6.015,19]henicosa-12,14,16,19-tetraen-18-one (20 mg, 44.9 μmol, 1 eq) in THF (0.6 mL) and H₂O (0.2 mL) was added NaHCO₃ (11 .3 mg, 134.7 μmol, 5.2 μL, 3 eq) and prop-2-enoyl chloride (4.1 mg, 44.9 μmol, 3.7 μL, 1 eq) at 0°C. The mixture was stirred at 0°C for 30 min. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (TFA condition, column: Phenomenex luna C18 100 x 40 mm x 5 um; mobile phase: [H₂O (0.1% TFA)-ACN]; gradient: 15%-45% B over 8.0 min) to give 17-(5-methyl-4-prop-2-enoyl-2,3-dihydroquinoxalin-1-yl)-1 , 6,11 ,13,20- pentazatetracyclo[10.6.2.12,6.015,19]henicosa-12,14,16,19-tetraen-18-one (11.2 mg, 21.08 μmol, 46.96% yield, 94.02% purity) as a yellow solid.

VT ¹H NMR (400 MHz, DMSO-d₆) δ = 8.56 (s, 1 H), 7.45 (s, 1 H), 6.76 (br dd, J = 10.6, 16.5 Hz, 1 H), 6.51 (br d, J = 7.0 Hz, 1 H), 6.39 (br t, J = 7.5 Hz, 1H), 6.27 (br d, J = 7.7 Hz, 1 H), 6.09 (dd, J = 2.3, 16.8 Hz, 1H), 5.64 (dd, J = 2.1 , 10.6 Hz, 1 H), 5.39 - 5.22 (m, 1 H), 4.42 - 3.99 (m, 3H), 3.62 - 3.59 (m, 4H), 3.41 (br s, 4H), 2.93 (br dd, J = 3.0, 12.0 Hz, 1H), 2.11 (s, 3H), 1.98 (br t, J = 6.6 Hz, 5H), 1.89 (br d, J = 12.9 Hz, 1 H), 1.82 - 1.75 (m, 1 H), 1.59 - 1.46 (m, 1 H).

Procedure for Preparation of Compound 001

Scheme 21

To a solution of (Z)-but-2-ene-1 ,4-diol (4 g, 45.4 mmol, 3.73 mL, 1 eq) and isoindoline-1 ,3-dione (6.7 g, 45.4 mmol, 1 eq) in THF (80 mL) was added PPh₃ (17.8 g, 68.1 mmol, 1 .5 eq). Then the mixture was added DIAD (10.1 g, 49.9 mmol, 9.7 mL, 1.1 eq) at 0°C. The mixture was stirred at 25°C for 12 hr. TLC indicated one new spot formed. The residue was diluted with H₂O (60 mL) and extracted with ethyl acetate (70 mL x 4), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (40 g Silica Flash Column, Eluent of 0-30% Ethyl acetate/Petroleum ether gradient @ 120 mL/min) to give 2-[(Z)-4-hydroxybut-2- enyl]isoindoline-1 ,3-dione (19 g, 25.3 mmol, 55.7% yield, 28.9% purity) as a light yellow solid.

To a solution of 2-[(Z)-4-hydroxybut-2-enyl]isoindoline-1 ,3-dione (7 g, 32.2 mmol, 1 eq) in EtOH (70 mL) was added N₂H₄.H₂O (4.9 g, 96.7 mmol, 4.8 mL, 98% purity, 3 eq). The mixture was stirred at 80°C for 1 hr. TLC indicated one new spot formed. The reaction mixture filtered and concentrated under reduced pressure to give (Z)-4-aminobut-2-en-1-ol (4.5 g, crude) as a light yellow solid.

To a solution of 4-chloro-2-methylsulfanyl-pyrimidine-5-carbaldehyde (4.7 g, 25.0 mmol, 1 eq) in MeCN (50 mL) was added tert-butyl 3-aminopiperidine-1-carboxylate (5 g, 25.0 mmol, 1 eq) and DIEA (9.7 g, 74.9 mmol, 13.1 mL, 3 eq). The mixture was stirred at 25°C for 2 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by flash silica gel chromatography (40 g Silica Flash Column, Eluent of 0-10% Ethyl acetate/Petroleum ether gradient @ 150 mL/min) to give tert-butyl 3-[(5-formyl-2- methylsulfanyl-pyrimidin-4-yl)amino]piperidine-1-carboxylate (5.9 g, 16.8 mmol, 67.2% yield) as a yellow oil.

To a solution of tert-butyl 3-[(5-formyl-2-methylsulfanyl-pyrimidin-4- yl)amino]piperidine-1-carboxylate (5.9 g, 16.8 mmol, 1 eq) in DMF (60 mL) was added Cs₂CO₃ (16.4 g, 50.3 mmol, 3 eq) and benzyl 4-(2-ethoxy-2-oxo-ethyl)-8-methyl-2,3- dihydroquinoxaline-1 -carboxylate (6.2 g, 16.8 mmol, 1 eq). The mixture was stirred at 40°C for 4hr. LC-MS showed desired compound was detected. The residue was diluted with H₂O (150 mL) and extracted with ethyl acetate (200 mL x 4). The combined organic layers were washed with brine (200 mL x 1), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (80 g Silica Flash Column, Eluent of 0-10% Ethyl acetate/Petroleum ether gradient @ 150 mL/min) to give benzyl 4-[8-(1-tert-butoxycarbonyl-3-piperidyl)-2-methylsulfanyl-7-oxo- pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (8 g, 12.2 mmol, 72.6% yield) as a yellow solid.

To a solution of benzyl 4-[8-(1-tert-butoxycarbonyl-3-piperidyl)-2-methylsulfanyl-7- oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (8 g, 12.2 mmol, 1 eq) in DCM (80 mL) was added m-CPBA (3.7 g, 18.3 mmol, 85% purity, 1.5 eq) at 0°C. The mixture was stirred at 25°C for 2 hr. LC-MS showed desired compound was detected.

The residue was diluted with Na₂SO₃ (70 mL) and extracted with dichloromethane (100 mL x 3). The combined organic layers were washed with NaHCO₃ (80 mL x 3), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give benzyl 4-[8-( 1 -tert- butoxycarbonyl-3-piperidyl)-2-methylsulfinyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3- dihydroquinoxaline-1 -carboxylate (8 g, crude) as a yellow solid.

To a solution of benzyl 4-[8-(1-tert-butoxycarbonyl-3-piperidyl)-2-methylsulfinyl-7-oxo- pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (6 g, 8.9 mmol, 1 eq) in Tol. (60 mL) was added DIEA (3.5 g, 26.8 mmol, 4.7 mL, 3 eq) and (Z)-4-aminobut-2- en-1-ol (2.4 g, 27.6 mmol, 3.1 eq). The mixture was stirred at 25°C for 1 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by flash silica gel chromatography (80 g Silica Flash Column, Eluent of 0-80% Ethyl acetate/Petroleum ether gradient @ 150 mL/min) to give benzyl 4-[8-(1-tert-butoxycarbonyl-3-piperidyl)-2-[[(Z)-4-hydroxybut-2- enyl]amino]-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (2 g, 2.9 mmol, 32.2% yield) as a yellow solid.

To a solution of benzyl 4-[8-(1-tert-butoxycarbonyl-3-piperidyl)-2-[[(Z)-4-hydroxybut-2- enyl]amino]-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (2 g, 2.9 mmol, 1 eq) in DCM (20 mL) was added TEA (1 .5 g, 14.4 mmol, 2.0 mL, 5 eq) and DMAP (35.1 mg, 287.4 μmol, 0.1 eq) and TosCI (1 .6 g, 8.6 mmol, 3 eq) at 0°C. The mixture was stirred at 25°C for 2 hr. LC-MS showed desired compound was detected. The residue was diluted with NaHCO₃ (40 mL) and extracted with dichloromethane (50 mL x 3), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (20 g Silica Flash Column, Eluent of 0-36% Ethyl acetate/Petroleum ether gradient @ 100 mL/min) to give benzyl 4-[8-( 1 -tert- butoxycarbonyl-3-piperidyl)-2-[[(Z)-4-chlorobut-2-enyl]amino]-7-oxo-pyrido[2,3-d]pyrimidin-6- yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (990 mg, 1.4 mmol, 48.2% yield) as a yellow solid.

A mixture of benzyl 4-[8-(1-tert-butoxycarbonyl-3-piperidyl)-2-[[(Z)-4-chlorobut-2- enyl]amino]-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (990 mg, 1 .4 mmol, 1 eq) in HCI/EtOAc (10 mL, 4M) and EtOAc (10 mL) was stirred at 25°C for 1 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent to give benzyl 4-[2-[[(Z)-4- chlorobut-2-enyl]amino]-7-oxo-8-(3-piperidyl)pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3- dihydroquinoxaline-1 -carboxylate (990 mg, crude, HCI) as a yellow solid.

To a solution of benzyl 4-[2-[[(Z)-4-chlorobut-2-enyl]amino]-7-oxo-8-(3- piperidyl)pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (990 mg, 1 .5 mmol, 1 eq, HCI) in DMF (18 mL) was added K₂CO₃ (630.9 mg, 4.6 mmol, 3 eq) and Nal (228.1 mg, 1.5 mmol, 1 eq). The mixture was stirred at 80°C for 2 hr. LC-MS showed desired compound was detected. The residue was diluted with H₂O (15 mL) and extracted with ethyl acetate (25 mL x 4). The combined organic layers were washed with brine (20 mL x 1 ), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (20 g Silica Flash Column, Eluent of 0-40% Methanol/Dichloromethane ether gradient @ 100 mL/min). The residue was purified by prep-HPLC (TFA condition, column: Phenomenex luna C18 100 x 40 mm x 5 um; mobile phase: [H₂O (0.1% TFA)-ACN]; gradient:25%-55% B over 8.0 min) to give benzyl 8-methyl-4-[(8Z)-18-oxo-1 ,6,11 ,13,20-pentazatetracyclo[10.6.2.12,6.015,19]henicosa- 8,12,14,16,19-pentaen-17-yl]-2,3-dihydroquinoxaline-1-carboxylate (280 mg, 484.70 μmol, 31 .85% yield) as a yellow solid.

A mixture of benzyl 8-methyl-4-[(8Z)-18-oxo- 1 ,6,11 ,13,20- pentazatetracyclo[10.6.2.12,6.015,19]henicosa-8, 12,14,16,19-pentaen- 17-y l]-2 , 3- dihydroquinoxaline-1-carboxylate (30 mg, 51.9 μmol, 1 eq) in HBr/AcOH(1 Ml, 33% purity) was stirred at 25°C for 1 hr. LC-MS showed desired compound was detected. The reaction mixture was quenched by addition NaHCO₃ (7 mL) at 0 °C, and extracted with ethyl acetate (10 mL x 3), dried over Na2SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (TFA condition column: Phenomenex luna C18 100 x 40 mm x 5 um; mobile phase: [H₂O(0.1 % TFA)-ACN]; gradient: 1%-27% B over 8.0 min) to give (8Z)-17-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-1 , 6, 11 ,13,20- pentazatetracyclo[10.6.2.12,6.015,19]henicosa-8,12,14,16,19-pentaen-18-one (12 mg, 27.1 μmol, 52.1% yield) as a yellow solid.

To a solution of (8Z)-17-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-1 ,6,11 ,13,20- pentazatetracyclo[10.6.2.12,6.015,19]henicosa-8,12,14,16,19-pentaen-18-one (12 mg, 27.1 μmol, 1 eq) in DCM (1 mL) was added TEA (8.2 mg, 81 .2 μmol, 11.3 μL, 3 eq) and prop-2- enoyl chloride (2.5 mg, 27.1 μmol, 2.2 μL, 1 eq) at 0°C. The mixture was stirred at 0°C for 0.5 hr. LC-MS showed 83% Reactant 1 was remained, then the mixture was added prop-2- enoyl chloride (3.7 mg, 40.6 μmol, 3.3 μL, 1.5 eq) at 0°C. The mixture was stirred at 0°C for 0.5 hr. LC-MS showed 32.5% Reactant 1 was remained, then the mixture was added TEA (8.2 mg, 81.2 μmol, 11.3 μL, 3 eq) at 0°C. The mixture was stirred at 0°C for 0.5 hr. LC-MS showed 19% Reactant 1 was remained, then the mixture was added prop-2-enoyl chloride (1 .2 mg, 13.5 μmol, 1 .1 μL, 0.5 eq) at 0°C. The mixture was stirred at 0°C for 0.5 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-HPLC (TFA condition, column: Phenomenex luna C 18 100 x 40 mm x 5 um; mobile phase: [H₂O (0.1% TFA)-ACN]; gradient: 10%-40% B over 8.0 min) to give (8Z)-17-(5-methyl-4-prop-2-enoyl-2,3- d i hyd roq u inoxal i n- 1 -y I )- 1 ,6,11,13,20-pentazatetracyclo[10.6.2.12,6.015,19]henicosa- 8,12,14,16,19-pentaen-18-one (8.19 mg, 13.36 μmol, 49.40% yield, 99.8% purity, TFA) as a yellow solid.

VT ¹H NMR (400 MHz, DMSO-d₆) δ= 10.38-9.67 (m, 1H), 8.58 (s, 1H), 8.16-7.96 (m, 1H), 7.85 - 7.67 (m, 1H), 6.96 - 6.80 (m, 1H), 6.70 - 6.56 (m, 1H), 6.43 - 6.33 (m, 1H), 6.32-6.11 (m, 2H), 5.81 - 5.53 (m, 4H), 5.01 (brt, J= 12.6 Hz, 1H), 4.96-4.69 (m, 1H), 4.65-4.51 (m, 1H),3.64 (brd, J=13.1 Hz, 2H), 3.56 - 3.53 (m, 1H),3.40 (brs, 2H), 3.31 - 3.25 (m, 2H), 3.13 - 2.88 (m, 2H), 2.30 - 2.18 (m, 1H), 2.12 (s, 3H), 2.10-1.80 (m, 3H).

Procedure for Preparation of Compound 037

Scheme 22

To a solution of benzyl 4-[8-(1-tert-butoxycarbonyl-3-piperidyl)-2-methylsulfinyl-7-oxo- pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (200 mg, 297.3 μmol, 1 eq) in Tol. (2 mL) was added 2-(2-aminoethoxy)ethanol (46.9 mg, 445.9 μmol, 44.7 μL, 1.5 eq) and DIEA (192.1 mg, 1.5 mmol, 258.9 μL, 5 eq). The mixture was stirred at 20°C for 12 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by flash silica gel chromatography (20 g Silica Flash Column, Eluent of 0-74% Ethyl acetate/Petroleum ether gradient @ 120 mL/min) to give benzyl 4-[8-(1-tert-butoxycarbonyl- 3-piperidyl)-2-[2-(2-hydroxyethoxy)ethylamino]-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl- 2,3-dihydroquinoxaline-1-carboxylate (650 mg, 716.5 μmol, 48.2% yield, 78.7% purity) as a yellow solid.

To a solution of benzyl 4-[8-(1-tert-butoxycarbonyl-3-piperidyl)-2-[2-(2- hydroxyethoxy)ethylamino]-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3- dihydroquinoxaline-1-carboxylate (850 mg, 1 .2 mmol, 1 eq) in DCM (9 mL) was added TEA (361.5 mg, 3.6 mmol, 497.2 μL, 3 eq), DMAP (14.6 mg, 119.1 μmol, 0.1 eq) and TosCI (454.0 mg, 2.4 mmol, 2 eq) at 0°C. The mixture was stirred at 20°C for 12 hr. LC-MS showed desired compound was detected. The residue was diluted with NaHCO₃ (20 mL) and extracted with Dichloromethane (20 mL x 3), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (12 g Silica Flash Column, Eluent of 0-50% Ethyl acetate/Petroleum ether gradient @ 120 mL/min) to give benzyl 4-[8-(1-tert-butoxycarbonyl- 3-piperidyl)-7-oxo-2-[2-[2-(p-tolylsulfonyloxy)ethoxy]ethylamino]pyrido[2,3-d]pyrimidin-6-yl]-8- methyl-2,3-dihydroquinoxaline-1 -carboxylate (734 mg, 605.9 μmol, 50.9% yield, 71.7% purity) as a yellow solid.

A mixture of benzyl 4-[8-(1-tert-butoxycarbonyl-3-piperidyl)-7-oxo-2-[2-[2-(p- tolylsulfonyloxy)ethoxy]ethylamino]pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3- dihydroquinoxaline-1-carboxylate (734 mg, 845.6 μmol, 1 eq) in HCI/EtOAc (7.5 mL, 4M) was stirred at 20°C for 1 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent, to give benzyl 8- methyl-4-[7-oxo-8-(3-piperidyl)-2-[2-[2-(p-tolylsulfonyloxy)ethoxy]ethylamino]pyrido[2,3- d]pyrimidin-6-yl]-2,3-dihydroquinoxaline-1 -carboxylate (800 mg, crude, HCI) as a yellow solid.

To a solution of benzyl 8-methyl-4-[7-oxo-8-(3-piperidyl)-2-[2-[2-(p- tolylsulfonyloxy)ethoxy]ethylamino]pyrido[2,3-d]pyrimidin-6-yl]-2,3-dihydroquinoxaline-1- carboxylate (650 mg, 808.1 μmol, 1 eq, HCI) in ACN (7 mL) was added TEA (408.9 mg, 4.0 mmol, 562.4 μL, 5 eq). The mixture was stirred at 60°C for 12 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by flash silica gel chromatography (12 g Silica Flash Column, Eluent of 0-25% Ethyl acetate/Petroleum ether gradient @ 120 mL/min) to give benzyl 8-methyl-4-(19-oxo-9-oxa-1 ,6,12,14,21- pentazatetracyclo[11 .6.2.12,6.016,20]docosa-13,15,17,20-tetraen-18-y l)-2, 3- dihydroquinoxaline-1 -carboxylate (60 mg, 75.5 μmol, 9.3% yield, 75.0% purity) as a yellow solid.

To a solution of Pd(OH)₂ (30 mg, 20% purity) in MeOH (10 mL) was added benzyl 8- methyl-4-(19-oxo-9-oxa-1 ,6,12,14,21-pentazatetracyclo[11 .6.2.12,6.016,20]docosa-

13.15.17.20-tetraen-18-yl)-2,3-dihydroquinoxaline-1-carboxylate (60 mg, 100.7 μmol, 1 eq) and TEA (10.2 mg, 100.7 μmol, 14.0 μL, 1 eq) under N₂ atmosphere. The suspension was degassed and purged with H₂ for 3 times. The mixture was stirred under H₂ (15Psi) at 20°C for 1 hr. LC-MS showed desired compound was detected. The residue was filtered and concentrated under reduced pressure to give a residue, to give 18-(5-methyl-3,4-dihydro-2H- quinoxalin-1-yl)-9-oxa-1 ,6,12,14,21-pentazatetracyclo[11 .6.2.12,6.016, 20]docosa-

13.15.17.20-tetraen-19-one (50 mg, crude) as a yellow solid.

To a solution of 18-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-9-oxa-1 ,6,12,14,21- pentazatetracyclo[11 ,6.2.12,6.016,20]docosa-13,15,17,20-tetraen-19-one (50 mg, 108.3 μmol, 1 eq) in DCM (1 mL) was added TEA (32.9 mg, 325.0 μmol, 45.2 μL, 3 eq) and prop- 2-enoyl chloride (5.9 mg, 65.0 μmol, 5.28 μL, 0.6 eq) at 0°C. The mixture was stirred at 0°C for 1 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep- HPLC (TFA condition column: Phenomenex luna C18 80 x 40 mm x 3 um; mobile phase: [H₂O(0.1% TFA)-ACN]; gradient: 10% - 45% B over 8.0 min) to give 18-(5-methyl-4-prop-2- enoyl-2,3-dihydroquinoxalin-1 -yl)-9-oxa-1 ,6,12,14,21- pentazatetracyclo[11 ,6.2.12,6.016,20]docosa-13,15,17,20-tetraen-19-one (6.2 mg, 11.99 μmol, 11 .07% yield, 99.71 % purity) as a yellow solid. Procedure for Preparation of Compound 004

Scheme 23

To a solution of benzyl 4-[8-(1-tert-butoxycarbonyl-3-piperidyl)-2-methylsulfinyl-7-oxo- pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (400 mg, 594.5 μmol, 1 eq) in Tol. (4 mL) was added DIPEA (230.5 mg, 1 .8 mmol, 310.7 μL, 3 eq) and tertbutyl 3-(2-aminoethoxy)propanoate (135.0 mg, 713.4 μmol, 1.2 eq). The mixture was stirred at 100°C for 12 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by flash silica gel chromatography (12 g Silica Flash Column, Eluent of 0-29% Ethyl acetate/Petroleum ether gradient @ 80 mL/min) to give benzyl 4-[8-(1-tert-butoxycarbonyl-3- piperidyl)-2-[2-(3-tert-butoxy-3-oxo-propoxy)ethylamino]-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8- methyl-2,3-dihydroquinoxaline-1-carboxylate (440 mg, 551.4 μmol, 92.8% yield) as a yellow solid.

To a solution of benzyl 4-[8-(1-tert-butoxycarbonyl-3-piperidyl)-2-[2-(3-tert-butoxy-3- oxo-propoxy)ethylamino]-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline- 1-carboxylate (390 mg, 488.8 μmol, 1 eq) in DCM (3 mL) was added TFA (1.5 g, 13.5 mmol, 1 mL, 27.5 eq). The mixture was stirred at 25°C for 2 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent to give 3-[2-[[6-(4-benzyloxycarbonyl-5-methyl-2,3-dihydroquinoxalin-1-yl)-7-oxo-8- (3-piperidyl)pyrido[2,3-d]pyrimidin-2-yl]amino]ethoxy]propanoic acid (300 mg, crude) as a brown solid.

To a solution of 3-[2-[[6-(4-benzyloxycarbonyl-5-methyl-2,3-dihydroquinoxalin-1-yl)-7- oxo-8-(3-piperidyl)pyrido[2,3-d]pyrimidin-2-yl]amino]ethoxy]propanoic acid (300 mg, 467.5 μmol, 1 eq) in DMF (30 mL) was added PyBOP (608.2 mg, 1 .2 mmol, 2.5 eq) and TEA (7.3 g, 71.8 mmol, 10 mL, 153.7 eq). The mixture was stirred at 25°C for 2 hr. LC-MS showed desired compound was detected. The residue was diluted with H₂O 100 mL and extracted with ethyl acetate (200 mL x 3). The combined organic layers were washed with brine (50 mL x 2), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiO₂, Dichloromethane : Methanol = 10:1 ) to give benzyl 4-(7,20-dioxo-10-oxa-1 ,6,13,15,22- pentazatetracyclo[12.6.2.12,6.017,21]tricosa-14,16,18,21-tetraen-19-yl)-8-methyl-2,3- dihydroquinoxaline-1-carboxylate (220 mg, 352.7 μmol, 75.4% yield) as yellow oil.

To a solution of Pd(OH)₂/C (100 mg, 20% purity) in MeOH (5 mL) was added benzyl 4-(7,20-dioxo-10-oxa-1 ,6,13,15,22-pentazatetracyclo[12.6.2.12,6.017,21 ]tricosa- 14, 16,18,21 -tetraen-19-yl)-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (120 mg, 192.4 μmol, 1 eq) under N₂ atmosphere. The suspension was degassed and purged with H₂ for 3 times. The mixture was stirred under H₂ (15 Psi) at 25°C for 1 hr. LC-MS showed desired compound was detected.

The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (TFA condition, column: Phenomenex Luna C 18 100 x 40 mm x 5 um; mobile phase: [H₂O (0.1% TFA) - ACN]; gradient: 7%-37% B over 8.0 min) to give 19-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-10-oxa-1 ,6,13,15,22- pentazatetracyclo[12.6.2.12,6.017,21]tricosa-14,16,18,21-tetraene-7,20-dione (80 mg, 163.4 μmol, 84.9% yield) as a yellow solid.

To a solution of 19-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)-10-oxa-1 ,6,13,15,22- pentazatetracyclo[12.6.2.12,6.017,21]tricosa-14,16,18,21-tetraene-7,20-dione (60 mg, 122.6 μmol, 1 eq) in DCM (1 mL) was added TEA (37.2 mg, 367.7 μmol, 51.2 μL, 3 eq) and prop- 2-enoyl chloride (11 .1 mg, 122.6 μmol, 10.0 μL, 1 eq) at 0°C. The mixture was stirred at 0°C for 1 hr. LC-MS showed desired compound was detected. The reaction mixture was concentrated under N₂ to remove solvent. The residue was purified by prep-HPLC (TFA condition, column: Phenomenex Luna C 18 100 x 40 mm x 5 um; mobile phase: [H₂O (0.1 % TFA) - ACN]; gradient: 20%-50% B over 8.0 min) to give 19-(5-methyl-4-prop-2-enoyl-2,3- dihydroquinoxalin-1 -yl)-10-oxa-1 ,6, 13, 15,22-pentazatetracyclo[12.6.2.12,6.017,21 ]tricosa- 14, 16, 18, 21-tetraene-7, 20-dione (56.84 mg, 103.9 μmol, 84.7% yield, 99.33% purity) as a yellow solid.

Procedure of Compound 003

To a solution of 4,4-difluorocyclohexanone (5 g, 37.3 mmol, 1 eq) in DCM (50 mL) was added m-CPBA (9.8 g, 48.4 mmol, 85% purity, 1 .3 eq) at 0°C. The mixture was stirred at 25°C for 96 hr. TLC plate indicated reactant 1 was consumed completely and one new spot formed. The reaction was clean according to TLC. The reaction mixture was partitioned between Na₂S03 (100 mL) and DCM (100 mL). The organic phase was separated, washed with NaHCO₃ (100 mL * 3), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give the crude product 5,5-difluorooxepan-2-one (5.6 g, crude) obtained as pale yellow solid used into the next step without further purification.

A mixture of 5,5-difluorooxepan-2-one (5.6 g, 37.3 mmol, 1 eq) in NH₃/MeOH (50 mL, 7M) was stirred at 25°C for 12 hr. TLC plate indicated reactant 1 was consumed completely and two new spots formed. The reaction was clean according to TLC. The reaction mixture was concentrated under reduced pressure to give the crude product 4,4-difluoro-6-hydroxy- hexanamide (6 g, crude) obtained as yellow solid used into the next step without further purification.

To a solution of 4,4-difluoro-6-hydroxy-hexanamide (6 g, 35.9 mmol, 1 eq) in THF (60 mL) was added LiAIH₄ (2.5 M, 28.7 mL, 2 eq) at 0°C. The mixture was stirred at 25°C for 12 hr. TLC plate indicated one new spot formed. The reaction was clean according to TLC. The reaction mixture was quenched by addition 3 mL MgSO₄ and 3 g MgSO₄ at 0°C, and then the mixture was filtered. The filtrate was concentrated under reduced pressure to give the crude product 6-amino-3,3-difluoro-hexan-1-ol (5 g, crude) obtained as yellow solid used into the next step without further purification.

To a solution of 4-chloro-2-methylsulfanyl-pyrimidine-5-carbaldehyde (4.3 g, 22.8 mmol, 1 eq) in MeCN (20 mL) was added DIEA (2.9 g, 22.8 mmol, 3.9 mL, 1 eq) and 6- amino-3,3-difluoro-hexan-1-ol (3.5 g, 22.8 mmol, 1 eq) .The mixture was stirred at 25°C for 12 hr. The reaction was monitored by LCMS which showed complete consumption of reactant and detection of the desired mass peak. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (40 g Silica Flash Column, Eluent of 0-100% Ethyl acetate/Petroleum ether gradient @ 120 mL/min) to yield compound 4-[(4,4-difluoro-6-hydroxy-hexyl)amino]-2- methylsulfanyl-pyrimidine-5-carbaldehyde (1.5 g, 4.9 mmol, 21.5% yield) obtained as a brown solid.

To a solution of 4-[(4,4-difluoro-6-hydroxy-hexyl)amino]-2-methylsulfanyl-pyrimidine- 5-carbaldehyde (1.2 g, 4.0 mmol, 1 eq) in DMF (20 mL) was added Cs₂CO₃ (3.9 g, 12.2 mmol, 3 eq) and benzyl 4-(2-ethoxy-2-oxo-ethyl)-8-methyl-2,3-dihydroquinoxaline-1- carboxylate (1.5 g, 4.0 mmol, 1 eq). The reaction mixture was stirred at 25°C for 12 hr. The reaction was monitored by LCMS which showed complete consumption of reactant and detection of the desired product. The reaction mixture was partitioned between aqueous NaCI (20 mL) and ethyl acetate (100 mL). The organic phase was separated, washed with aqueous NaCI (15 mL * 1 ), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (20 g Silica Flash Column, Eluent of 0-100% Ethyl acetate/Petroleum ether gradient @ 50 mL/min) to give compound benzyl 4-[8-(4,4-difluoro-6-hydroxy-hexyl)-2-methylsulfanyl-7- oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1-carboxylate (800 mg, 1.3 mmol, 32.1% yield) obtained as a yellow solid.

To a solution of benzyl 4-[8-(4,4-difluoro-6-hydroxy-hexyl)-2-methylsulfanyl-7-oxo- pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (600 mg, 984.1 μmol, 1 eq) in DCM (10 mL) was added m-CPBA (299.7 mg, 1.4 mmol, 85% purity, 1.5 eq) at 0°C. The mixture was stirred at 25°C for 12 hr. The reaction was monitored by LCMS which showed complete consumption of reactant and detection of the desired product. The reaction mixture was partitioned between Na₂SO₃ (10 mL) and DCM (30 mL). The organic phase was separated, washed with NaHCO₃ mL (10 mL * 3), dried over Na₂SO₄, filtered and concentrated under reduced pressure to the crude product benzyl 4-[8-(4,4-difluoro-6- hydroxy-hexyl)-2-methylsulfonyl-7-oxo-pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3- dihydroquinoxaline-1 -carboxylate (700 mg, crude) obtained as yellow solid used into the next step without further purification.

To a solution of 1-tetrahydropyran-2-ylpyrazol-4-amine (234.5 mg, 1.4 mmol, 3 eq) in i-PrOH (3 mL) was added benzyl 4-[8-(4,4-difluoro-6-hydroxy-hexyl)-2-methylsulfonyl-7-oxo- pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (300 mg, 467.5 μmol, 1 eq). The mixture was stirred at 60°C for 12 hr. The reaction was monitored by LCMS which showed complete consumption of reactant and detection of the desired product.

The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (12 g Silica Flash Column, Eluent of 0-100% Ethyl acetate/Petroleum ether gradient @ 100 mL/min) to give compound benzyl 4- [8-(4,4-difluoro-6-hydroxy-hexyl)-7-oxo-2-[(1-tetrahydropyran-2-ylpyrazol-4- yl)amino]pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (720 mg) obtained as a brown oil.

To a solution of benzyl 4-[8-(4,4-difluoro-6-hydroxy-hexyl)-7-oxo-2-[(1- tetrahydropyran-2-ylpyrazol-4-yl)amino]pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3- dihydroquinoxaline-1-carboxylate (580 mg, 795.8 μmol, 1 eq) in DCM (15 mL) was added TEA (241.5 mg, 2.3 mmol, 332.3 μL, 3 eq) and MsCI (136.7 mg, 1.1 mmol, 92.4 μL, 1.5 eq) at 0°C. The mixture was stirred at 25°C for 30 min. LCMS showed 30% the reactant 1 was remained and 40% desired mass was detected. MsCI (136.7 mg, 1.1 mmol, 92.4 μL, 1.5 eq) was added to the mixture at 0°C. The reaction mixture was stirred at 25°C for 30 min. The reaction was monitored by LCMS which showed complete consumption of reactant and detection of the desired product. The reaction mixture was quenched by addition NaHCO₃ (30 mL) at 0°C, and extracted with DCM (30 mL * 3). The combined organic layers were dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (20 g Silica Flash Column, Eluent of 0-100% Ethyl acetate/Petroleum ether gradient @ 100 mL/min) to give compound benzyl 4- [8-(4,4-difluoro-6-methylsulfonyloxy-hexyl)-7-oxo-2-[(1-tetrahydropyran-2-ylpyrazol-4- yl)amino]pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (230 mg, 285.0 μmol, 35.8% yield) obtained as a yellow solid.

To a solution of benzyl 4-[8-(4,4-difluoro-6-methylsulfonyloxy-hexyl)-7-oxo-2-[(1- tetrahydropyran-2-ylpyrazol-4-yl)amino]pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3- dihydroquinoxaline-1-carboxylate (230 mg, 285.0 μmol, 1 eq) in EtOAc (4 mL) was added HCI/EtOAc (4 M, 427.5 μL, 6 eq). The mixture was stirred at 25°C for 12 hr. The reaction was monitored by LCMS which showed complete consumption of reactant and detection of the desired product. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (TFA condition;column: Phenomenex luna C18 100 * 40 mm * 5 um; mobile phase: [H₂O(0.1% TFA)-ACN]; gradient: 35%-65% B over 8.0 min) to give the crude product benzyl 4-[8-(4,4-difluoro-6-methylsulfonyloxy-hexyl)- 7-oxo-2-(1 H-pyrazol-4-ylamino)pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline- 1 -carboxylate (200 mg, 276.7 μmol, 97.0% yield) obtained as brown solid.

To a solution of benzyl 4-[8-(4,4-difluoro-6-methylsulfonyloxy-hexyl)-7-oxo-2-(1 H- pyrazol-4-ylamino)pyrido[2,3-d]pyrimidin-6-yl]-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (140 mg, 193.7 μmol, 1 eq) in DMF (3 mL) was added K₂CO₃ (160.6 mg, 1.1 mmol, 6 eq). The mixture was stirred at 60°C for 18 hr. LC-MS showed 27% desired mass was detected and 35% reactant 1 was remained. The mixture was stirred at 80°C for 3 hr. The reaction was monitored by LCMS which showed complete consumption of reactant and detection of the desired product. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with H₂O (20 mL) and extracted with ethyl acetate (20 mL * 3). The combined organic layers were washed with brine (50 mL * 1 ), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiO₂, DCM: MeOH = 10:1 ) to give compound benzyl 4-(9,9- difluoro-14-oxo-2,5,6,13,19, 20-hexazatetracyclo[11 .6.2.13,6.017,21 ]docosa- 1 (19), 3(22), 4, 15,17, 20-hexaen-15-yl)-8-methyl-2,3-dihydroquinoxaline-1 -carboxylate (80 mg, 127.6 μmol, 65.9% yield) obtained as a yellow solid.

To a solution of Pd(OH)₂ (20 mg, 20% purity) in MeOH (1 mL) /EtOAc (1.2 mL) was added benzyl 4-(9,9-difluoro-14-oxo-2,5,6,13, 19,20- hexazatetracyclo[11 ,6.2.13,6.017,21]docosa-1 (19),3(22),4,15,17,20-hexaen-15-yl)-8-methyl- 2,3-dihydroquinoxaline-1-carboxylate (20 mg, 31.9 μmol, 1 eq) under N₂. The suspension was degassed and purged with H₂ (15 psi) for 3 times. The mixture was stirred at 25°C for 1 hr.. The reaction was monitored by LCMS which showed complete consumption of reactant and detection of the desired product. The reaction mixture was concentrated under reduced pressure to give the crude product 9,9-difluoro-15-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)- 2,5,6, 13,19, 20-hexazatetracyclo[11.6.2.13,6.017,21]docosa-1(19),3(22),4,15,17,20-hexaen- 14-one (40 mg, crude) obtained as yellow solid used into the next step without further purification.

To a solution of 9,9-difluoro-15-(5-methyl-3,4-dihydro-2H-quinoxalin-1-yl)- 2,5,6, 13,19, 20-hexazatetracyclo[11.6.2.13,6.017,21]docosa-1(19),3(22),4,15,17,20-hexaen- 14-one (17 mg, 34.5 μmol, 1 eq) in DCM (1 mL) was added DIEA (13.3 mg, 103.5 μmol, 18.0 μL, 3 eq) and prop-2-enoyl chloride (3.1 mg, 34.5 pmol, 2.8 μL, 1 eq). The mixture was stirred at 0°C for 3 hr. The reaction was monitored by LCMS which showed complete consumption of reactant and detection of the desired mass peak. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep- HPLC (TFA condition; column: Phenomenex luna C18 100*40 mm*5 um; mobile phase: [H₂O(0.1% TFA)-ACN]; gradient:20%-50% B over 8.0 min) to yield the crude product 9,9- difluoro-15-(5-methyl-4-prop-2-enoyl-2,3-dihydroquinoxalin-1-yl)-2,5,6,13,19,20- hexazatetracyclo[11 ,6.2.13,6.017,21]docosa-1 (19),3(22),4,15,17,20-hexaen-14-one (97.202% purity, 6 mg) obtained as yellow solid.

Characterization data are presented below in Table 2.

Table 2

Example 2: Ba/F3 cell proliferation models

The EGFR mutant L858R Ba/F3 cells have been previously described (Zhou, W., et al. Nature 462, 2009, 1070-1074). The EGFR C797S mutations were introduced via site directed mutagenesis using the Quick Change Site-Directed Mutagenesis kit into a vector containing EGFR L858R mutation (Stratagene; La Jolla, CA) according to the manufacturer's instructions. All constructs were confirmed by DNA sequencing. The constructs were then shuttled into the retroviral vector JP1540 by either using the Cre-recombination system (Agilent Technologies, Santa Clara, CA) or the In-fusion HD Cloning kit (Takara Bio USA, Inc.; Mountain view, CA). Ba/F3 cells were then infected with retrovirus per standard protocols, as described previously (Zhou, et al, Nature 2009). Stable clones were obtained by selection in puromycin (2 μg/ml). All BaF/3 mutant cells were maintained in RPMI 1640 (Cellgro; Mediatech Inc., Herndon, CA) supplemented with 10% FBS, 100 units/mL penicillin, 100 units/mL streptomycin. Growth and inhibition of growth was assessed by the Cell Titer Gio assay (Promega,

Madison, Wl) and was performed according to the manufacturer’s instructions. The Cell Titer Gio assay is a luminescence-based method used to determine the number of viable cells based on quantitation of the ATP present, which is directly proportional to the amount of metabolically active cells present. Ba/F3 cells of different EGFR genotypes were exposed to compounds for 72 hours and the number of cells used per experiment was determined empirically as has been previously established (Zhou, et al., Nature 2009). All experimental points were set up in triplicates in 384-well plates. The luminescent signal was detected using a spectrometer and the data was graphically displayed using GraphPad Prism version 5.0 for Windows, (GraphPad Software; www.graphpad.com). The curves were fitted using a non-linear regression model with a sigmoidal dose response.

Ba/F3 cells assay data of the test compounds are provided in Table 3 below. For inhibitory activity against EGFR mutants, the following designations are used: ≤ 0.1 μM = A, 0.1 μM~0.5 μM = B, 0.5 μM~2.5 μM = C, ≥ 2.5 μM = D.

Table 3.

Example 3: Intact Mass Spectrometry

EGFR L858R protein (10 μg) was treated with DMSO or a 10-fold molar excess of compound for 2 h at 37 °C and analyzed by LC-MS using an HPLC (Shimadzu, Marlborough, MA) interfaced to an LTQ ion trap mass spectrometer (ThermoFisher Scientific, San Jose, CA). Protein (5 μg) was injected onto a reversed phase column (5 cm POROS 50R2, Applied Biosystems, Foster City, CA), desalted for 4 min (100% A), and gradient eluted (0-100% B in 1 min; A = 0.2 M acetic acid in water; B = 0.2 M acetic acid in acetonitrile) into the mass spectrometer (spray voltage = 4.8 kV). The mass spectrometer was programmed to acquire profile mass spectra (m/z 300-2000). Raw data was deconvoluted using MagTran version 1.03b2. (Rao, S. et al. Cell chemical biology 2019, 26, 818- 829. e9) To determine which amino acid was modified by the inhibitor, labeled protein was first reduced (10 mM dithiothreitol for 30 min at 56 °C), alkylated (22.5 mM iodoacetamide for 30 min at room temperature and protected from light), and digested with trypsin. Peptides were then desalted by C18, dried by vacuum centrifugation, reconstituted in 50% MeCN, 1% formic acid, and 100 mM ammonium acetate, and analyzed by CE-MS using a ZipChip autosampler and CE system (908 Devices, Boston, MA) interfaced to a QE- HF mass spectrometer (Thermofisher Scientific). The mass spectrometer was operated in data-dependent mode, and the 5 most abundant ions in each MS scan (m/z 300-2000, resolution = 60K, target = 3 x 106, max fill time = 50 ms) were subjected to MS/MS (collision energy = 35%, target = 1 x 105, max fill time = 100 ms). Dynamic exclusion was enabled with a repeat count of 1 and an exclusion duration of 6 s. Raw data were converted to .mgf using the multiplier toolset (Zhu, K. et al. J. Chem. Inf. Model. 2014, 54, 1932- 1940) and searched using Mascot 2.6.1 against a forward reversed human refseq database (NCBI). Search parameters specified a precursor mass tolerance of 50 ppm, a product ion tolerance of 25 mmu, fixed carbamidomethylation of cysteine, and variable oxidation of methionine as well as variable compounds modification of cysteine. Inhibitor related fragment ions were assigned as described. (Clark, N. A. et al. BMC Cancer 2017, 17, 698)

Data from the intact MS labeling experiments are provided in Table 4 below. Table 4

The disclosed subject matter is not to be limited in scope by the specific embodiments and examples described herein. Indeed, various modifications of the disclosure in addition to those described will become apparent to those skilled in the art from the foregoing description and accompanying figures. Such modifications are intended to fall within the scope of the appended claims.

All references (e.g., publications or patents or patent applications) cited herein are incorporated herein by reference in their entirety and for all purposes to the same extent as if each individual reference (e.g., publication or patent or patent application) was specifically and individually indicated to be incorporated by reference in its entirety for all purposes. Other embodiments are within the following claims.

Claims

1. A compound of Formula (IA):

or a pharmaceutically acceptable salt thereof, wherein:

X¹ is N or CH;

X² is N, CH, or CR⁸;

R² is represented by Formula (i-1):

Y is O, S, or CH₂.

2. The compound of claim 1 , wherein the compound is a compound of Formula (I A-l ):

or a pharmaceutically acceptable salt thereof, wherein:

X¹ is N or CH;

R⁵ and R⁶ are each independently selected from the group consisting of H, halo, CN, OH, NH₂, N(H)(C_1-6 alkyl), N(C_1-6 alkyl)₂, C_1-6 alkyl, C_1-6 alkoxy, and C_1-6 haloalkyl; R⁹ is selected from the group consisting of halo, CN, OH, NH₂, N(H)(C_1-6 alkyl), N(C_1- ₆ alkyl)₂, C_1-6 alkyl, C_1-6 alkoxy, and C_1-6 haloalkyl; each n, p, and r is independently 0, 1 , or 2; s is 1-15; is a single bond, a cis double bond, or a trans double bond; and

R² is represented by Formula (i-1):

Y is O, S, or CH₂.

3. The compound of claim 1 or 2, wherein:

X¹ is N or CH;

R¹ is selected from the group consisting of halo, OH, C_1-6 alkyl, C_1-6 alkoxy, and C_1-6 haloalkyl;

R⁵ and R⁶ are each independently selected from the group consisting of H, halo, CN, OH, C_1-6 alkyl, C_1-6 alkoxy, and C_1-6 haloalkyl; each R⁹ is independently selected from the group consisting of CN, NH₂, N(H)(C_1-6 alkyl), and N(C_1-6 alkyl)₂; each n, p, and r is independently 0, 1 , or 2; s is 1-12; is a single bond, a cis double bond, or a trans double bond; and R² is represented by Formula (i-1 ):

Y is O, S, or CH₂.

4. The compound of any one of claims 1-3, wherein:

X¹ is N or CH;

R¹ is selected from the group consisting of C_1-6 alkyl, C_1-6 alkoxy, and C_1-6 haloalkyl;

R³ and R⁴ are each independently selected from the group consisting of C_1-6 alkyl, C_1- ₆ alkoxy, C_1-6 haloalkyl, and C_1-6 alkylcarbonyl, wherein C_1-6 alkyl, C_1-6 alkoxy, C_1-6 haloalkyl, and C_1-6 alkylcarbonyl are optionally substituted with one R⁹;

R² is represented by Formula (i-1 ):

wherein: L₃ is a bond, N(H), or N(C_1-6 alkyl); each R_E1 , R_E2, and R_E3 is independently selected from the group consisting of H, halo, and C_1-6 alkyl; and

Y is O, S, or CH₂.

5. The compound of any one of claims 1-4, wherein the compound is a compound of Formula (IA-II):

6. The compound of any one of claims 1-5, wherein Ring A and Ring B are each independently selected from the group consisting of absent, phenyl, 5- or 6-membered heterocycloalkyl, 5- or 6-membered heteroaryl, and 8- or 9-membered bicyclic ring.

7. The compound of any one of claims 1-6, wherein Ring A and Ring B are each independently selected from the group consisting of absent, phenyl, 6-membered heterocycloalkyl, 5- or 6-membered heteroaryl, and 9-membered bicyclic ring.

8. The compound of any one of claims 1-7, wherein the heterocycloalkyl, heteroaryl, and bicyclic ring of Ring A and Ring B contains 1 , 2, or 3 heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur.

9. The compound of any one of claims 1-8, wherein the heterocycloalkyl, heteroaryl, and bicyclic ring of Ring A and Ring B contains 1 or 2 heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur.

10. The compound of any one of claims 1-9, wherein Ring A is selected from the group consisting of absent, phenyl, 5- or 6-membered heteroaryl, and 9-membered bicyclic ring.

11 . The compound of any one of claims 1-10, wherein Ring B is selected from the group consisting of absent, 6-membered heterocycloalkyl, and 5-membered heteroaryl.

12. The compound of any one of claims 1-11 , wherein one of Ring A or Ring B is not absent.

13. The compound of any one of claims 1-11 , wherein the compound is a compound of Formula (lA-lla):

or a pharmaceutically acceptable salt thereof, wherein:

X³ is N or CH, provided that when E is bonded to X³, then X³ is C; each E is independently selected from the group consisting of a bond, CR⁵R⁶, O, C(O), and NR¹⁰;

14. The compound of any one of claims 1-11 , wherein the compound is a compound of Formula (lA-llb):

15. The compound of any one of claims 1-1 1 , wherein the compound is a compound of Formula (lA-llc):

or a pharmaceutically acceptable salt thereof, wherein: each X⁵ is independently NR¹¹ or CH₂, provided that when E is bonded to an X⁵, then said X⁵ is N or CH; each E is independently selected from the group consisting of a bond, CR⁵R⁶, O, C(O), and NR¹⁰; R¹⁰ is selected from the group consisting of H, C_1-6 alkyl, C_1-6 haloalkyl, and C_1-6 alkylcarbonyl;

16. The compound of any one of claims 1-12, wherein the compound is a compound of Formula (lA-llaa):

17. The compound of any one of claims 1-11 and 13, wherein the compound is a compound of Formula (lA-llba):

18. The compound of any one of claims 1-1 1 and 14, wherein the compound is a compound of Formula (lA-llca):

19. The compound of any one of claims 1-1 1 and 14, wherein the compound is a compound of Formula (lA-llcb):

20. The compound of any one of claims 1-18, wherein R¹ is C_1-6 alkyl.

21 . The compound of any one of claims 1-16 and 19, wherein R³ and R⁴ are each independently selected from the group consisting of C_1-6 alkyl, C_1-6 alkoxy, C_1-6 haloalkyl, and C_1-6 alkylcarbonyl, wherein C_1-6 alkoxy is optionally substituted with one R⁹.

22. The compound of any one of claims 1-16, 19, and 20, wherein R³ is selected from the group consisting of C_1-6 alkyl, C_1-6 alkoxy, and C_1-6 alkylcarbonyl, wherein C_1-6 alkoxy is optionally substituted with one R⁹.

23. The compound of any one of claims 1-21 , wherein R⁵ and R⁶ are each independently selected from the group consisting of H, halo, C_1-6 alkyl, and C_1-6 haloalkyl.

24. The compound of any one of claims 1-22, wherein L₃ is a bond.

25. The compound of any one of claims 1-23, wherein n is 1 .

26. The compound of any one of claims 1-24, wherein p is 0 or 1 .

27. The compound of any one of claims 1-25, wherein r is 0 or 1.

28. The compound of any one of claims 1-26, wherein Y is O.

29. The compound of any one of claims 5-27, wherein at least one E is a bond.

30. The compound of any one of claims 5-27, wherein at least one E is O.

31 . The compound of any one of claims 5-27, wherein at least two E are either O or

NR¹⁰.

32. The compound of any one of claims 5-28, wherein at least two E are a bond.

33. The compound of any one of claims 5-28 and 31 , wherein at least three E are a bond.

34. The compound of any one of claims 1-32, wherein the compound is selected from the

or a pharmaceutically acceptable salt thereof.

35. A pharmaceutical composition comprising a compound of any one of claims 1-33, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

36. A method of inhibiting the activity of EGFR in a subject in need thereof comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-33 or the pharmaceutical composition of claim 34.

37. A method of treating cancer in a subject in need thereof comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1-33 or the pharmaceutical composition of claim 34.

38. The method of claim 36, wherein the cancer is selected from the group consisting of lung cancer, colon cancer, breast cancer, endometrial cancer, thyroid cancer, glioma, squamous cell carcinoma, and prostate cancer.

39. The method according to claim 36, wherein the cancer is non-small cell lung cancer (NSCLC).