WO2023185864A1 - Composés pour la dégradation ciblée de kras - Google Patents

Composés pour la dégradation ciblée de kras Download PDF

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WO2023185864A1
WO2023185864A1 PCT/CN2023/084424 CN2023084424W WO2023185864A1 WO 2023185864 A1 WO2023185864 A1 WO 2023185864A1 CN 2023084424 W CN2023084424 W CN 2023084424W WO 2023185864 A1 WO2023185864 A1 WO 2023185864A1
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alkyl
optionally substituted
halogen
pharmaceutically acceptable
mmol
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PCT/CN2023/084424
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Ying Han
Sidney Siubun YU
Dapeng Li
Yao-Ling Qiu
Zhiyu Yin
Jinli Zhang
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Jingrui Biopharma Co., Ltd.
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Publication of WO2023185864A1 publication Critical patent/WO2023185864A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/545—Heterocyclic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/55—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug, i.e. a dimer, oligomer or polymer of pharmacologically or therapeutically active compounds
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/08—Bridged systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration

Definitions

  • the present invention is directed to novel compounds comprising a target protein binding moiety and an E3 ubiquitin ligase binding moiety via a linker, and associated methods of use.
  • Such compounds are useful as modulators of targeted ubiquitination of KRAS (Kirsten ras sarcoma) protein with a G12D mutation and/or G12V, which is then degraded and/or inhibited.
  • KRAS Kersten ras sarcoma
  • KRAS mutations cause oncogenic transformation of normal cells to cancerous cells.
  • One of the most frequently mutations in KRAS happens in codon 12, resulting in a change of glycine 12 to aspartate (i.e. G12D) , cysteine (G12C) , valine (G12V) and alanine (G12A) .
  • G12D is common in pancreatic ductual adenocarcinoma ( ⁇ 25%) , colorectal carcinoma ( ⁇ 12%) , and non-small cell lung carcinoma ( ⁇ 4%) .
  • G12D mutation increases the signaling activity of KRAS and overall activation of the EGF receptor signaling pathway, which plays a pivotal role in cell proliferation, to pathological level. Being an activating mutation, G12D happening in one allele is sufficient for oncogenic transformation, but both heterozygous and homozygous G12D mutation have been identified in cancer samples.
  • KRAS is an attractive target for therapeutic development.
  • small chemical inhibitor targeting KRAS G12C has only recently been approved by FDA.
  • the KRAS protein molecule does not offer a surface that accommodates small chemical compound deep within the protein with limited exposure of the inhibitory compound to surrounding environment.
  • Second, cellular concentration of GTP in mM range allows KRAS to bind GTP at high affinity. This increases the difficulty of finding a competitive GTP analog.
  • the proteasome recognizes and degrades cellular proteins that are conjugated with ubiquintin. This conjugation step is mediated by a family of ubiquitin ligases, among which are the E3 ligases. Increasingly, utilizing the ubiquitin E3 ligase to direct ubiquitin conjugation of a pathological protein target, leading to subsequent removal of such protein by intrinsic protein degradation machinery, is a viable approach of therapeutic intervention.
  • proteolysis targeting chimeras involves designing a bifunctional chemical compound that links up the intended protein target and an E3 ligase, and by so doing, brings the target protein close to the proteasome and induces protein degradation of the target, and thus removing the pathological enzymatic activity of the target protein within the affected cells.
  • the PROTAC bifunctional chemical compound is composed of a moiety that binds to the intended target, a linker, and a moiety that binds to an E3 ligase (or a subunit of it) .
  • the E3 ligase used in this disclosure is the Von Hippel Lindau (VHL) .
  • VHL is the substrate recognition subunit of an E3 ligase complex called VCB.
  • a small chemical molecule that binds to VHL has been incorporated in various PROTAC designs.
  • Other E3 ligases should also be effective if they are expressed in the tumor cells of intended treatment.
  • KRAS G12D Using PROTAC-mediated degradation of KRAS G12D is a plausible approach to intervene the oncogenic hyperactivity of KRAS G12D in cancer. By degrading KRAS G12D, KRAS signaling activity is reduced to a level that fails to support the growth of tumor. However, the research of KRAS G12D PROTAC compound has been progressing slowly for a long time.
  • the present disclosure describes compounds (i.e., hetero-bifunctional compounds) that function to recruit Kirsten ras sarcoma protein (KRas or KRAS) , such as mutant or gain-of-function KRas, to an E3 ubiquitin ligase for targeted ubiquitination and subsequent proteasomal degradation, and methods of making and using the same.
  • KRas or KRAS Kirsten ras sarcoma protein
  • a disease condition such as a KRas-related disease or disorder, e.g., accumulation or overactivity of an KRas protein or a mutated or gain-of function KRas protein or a mis-folded KRas protein, pancreatic cancer, colon cancer, colorectal cancer, lung cancer, non-small cell lung cancer, biliary tract malignancies, endometrial cancer, cervical cancer, bladder cancer, liver cancer, myeloid leukemia, breast cancer and brain cancer.
  • a KRas-related disease or disorder e.g., accumulation or overactivity of an KRas protein or a mutated or gain-of function KRas protein or a mis-folded KRas protein
  • pancreatic cancer colon cancer
  • colorectal cancer lung cancer, non-small cell lung cancer, biliary tract malignancies, endometrial cancer, cervical cancer, bladder cancer, liver cancer, myeloid leukemia, breast cancer and brain cancer.
  • the disclosure provides the compounds, which comprise an E3 ubiquitin ligase binding moiety and a moiety that binds KRas or a mutated version thereof, such that the KRas protein is thereby placed in proximity to the ubiquitin ligase to effect ubiquitination and subsequent degradation (and/or inhibition) of the KRas protein.
  • an E3 ubiquitin ligase binding moiety and a moiety that binds KRas or a mutated version thereof, such that the KRas protein is thereby placed in proximity to the ubiquitin ligase to effect ubiquitination and subsequent degradation (and/or inhibition) of the KRas protein.
  • P is a protein targeting moiety, ------is a bond, and U is an E3 ubiquitin ligase binding moiety.
  • the compound further comprises a chemical “Linker” .
  • the structure of the compound can be depicted as:
  • the compounds as described herein comprise multiple independently selected Ps, multiple Us, multiple chemical linkers or a combination thereof.
  • the P is a small molecule that binds KRas or a mutant thereof, such as a gain-of-function KRas.
  • the P is a small molecule that binds KRas. In any of the aspects or embodiments described herein, the P is a small molecule that binds both a KRas wild type protein and a KRas mutant, such as a KRas protein that has gain-of-function mutation.
  • the P is a small molecule that binds both a KRas wild type protein and a KRas mutant such as, but not limited to, a gain-of-function KRas mutant.
  • the small molecule that binds the KRas is as described herein.
  • the U is a derivative of trans-3-hydroxyproline, where both nitrogen and carboxylic acid in trans-3-hydroxyproline are functionalized as amides.
  • “Linker” is a bond.
  • the “Linker” is a connector with a linear non-hydrogen atom number in the range of 1 to 45 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44 or 45) .
  • the connector “Linker” can contain, but is not limited to one or more functional groups such as ether, amide, alkane, alkene, alkyne, ketone, hydroxyl, carboxylic acid, thioether, sulfoxide, and sulfone.
  • the linker can contain aromatic, heteroaromatic, cyclic, bicyclic or tricyclic moieties. Substitution with halogen, such as Cl, F, Br and I, or alkyl, such as methyl, ethyl, isopropyl, and tert-butyl, can be included in the linker. In the case of fluorine substitution, single or multiple fluorines can be included.
  • halogen such as Cl, F, Br and I
  • alkyl such as methyl, ethyl, isopropyl, and tert-butyl
  • the description provides therapeutic compositions comprising an effective amount of a compound as described herein, or a salt form thereof, and a pharmaceutically acceptable carrier.
  • the therapeutic compositions can be used to trigger targeted degradation of KRas or a mutated version thereof and/or inhibition of KRas or a mutated version thereof, in a patient or subject, for example, an animal such as a human, and can be used for treating or ameliorating one or more disease states, conditions, or symptoms causally related to KRas or mutated version thereof, which treatment is accomplished through degradation or inhibition of the KRas protein or mutated version thereof, or controlling or lowering KRas protein levels or protein levels of a mutated version thereof, in a patient or subject.
  • the therapeutic compositions as described herein may be used to effectuate the degradation of KRas, or a mutant or mis-folded form thereof, for the treatment or amelioration of a disease such as, e.g., accumulation, aggregation, or overeactivity of a KRas protein, a mis-folded, or a mutated form thereof (such as a gain-of-function KRas protein, pancreatic cancer, colon cancer, colorectal cancer, lung cancer, non-small cell lung cancer, biliary tract malignancies, endometrial cancer, cervical cancer, bladder cancer, liver cancer, myeloid leukemia, breast cancer and brain cancer) .
  • a disease such as, e.g., accumulation, aggregation, or overeactivity of a KRas protein, a mis-folded, or a mutated form thereof (such as a gain-of-function KRas protein, pancreatic cancer, colon cancer, colorectal cancer, lung cancer, non
  • the present disclosure provides a method of ubiquitinating KRas or a mutated form thereof in a cell (e.g., in vitro or in vivo) .
  • the method comprises administering a hetero-bifunctional compound as described herein comprising a P that binds KRas or a mutant form thereof, and a U, preferably linked through a chemical linker moiety, as described herein, to effectuate degradation of the KRas protein or mutant form thereof.
  • the control or reduction in levels of the KRas protein or mutated form thereof afforded by the present disclosure provides treatment of a KRas causally related disease state, condition or related symptom, as modulated through a lowering of the amount of KRas protein or mutated form thereof in cells of the subject.
  • the description provides methods for treating or ameliorating a disease, condition, or symptom thereof causally related to KRas or mutated form thereof in a subject or a patient, e.g., an animal such as a human, comprising administering to a subject in need thereof a composition comprising an effective amount, e.g., a therapeutically effective amount, of a hetero-bifunctional compound as described herein or salt form thereof, and a pharmaceutically acceptable carrier, wherein the composition is effective for treating or ameliorating the disease or disorder or symptom thereof in the subject.
  • a composition comprising an effective amount, e.g., a therapeutically effective amount, of a hetero-bifunctional compound as described herein or salt form thereof, and a pharmaceutically acceptable carrier, wherein the composition is effective for treating or ameliorating the disease or disorder or symptom thereof in the subject.
  • the method further includes, prior to administering a composition or compound of the present disclosure to a subject, identifying a patient as having a mutant KRas protein (e.g., KRas G12D, G12V) .
  • a mutant KRas protein e.g., KRas G12D, G12V
  • the description provides methods for identifying the effects of the degradation of KRas protein in a biological system using compounds according to the present disclosure.
  • the description provides processes and intermediates for making a hetero-bifunctional compound of the present disclosure capable of targeted ubiquitination and degradation of the KRas protein in a cell (e.g., in vivo or in vitro) .
  • Figures 1A and 1B Degradation of KRAS G12D by example compounds 1 to 5 in SW1990 cells; (1A) KRAS protein level was determined by probing with KRAS specific antibody (upper panels) . Immunoblotting with beta-Actin serves as loading control (lower panels) ; and (1B) Quantification of the relative abundance of the KRAS G12D signals as a ratio of the corresponding abundance of beta-Actin in the same samples (also see Table 1) .
  • FIGS 2A and 2B Degradation of KRAS G12D by example compounds 1 to 5 in AsPC-1 cells; KRAS protein level was determined by probing with KRAS specific antibody (upper panels) . Immunoblotting with beta-Actin serves as loading control (lower panels) ; and (1B) Quantification of the relative abundance of the KRAS G12D signals as a ratio of the corresponding abundance of beta-Actin in the same samples (also see Table 1) .
  • FIG. 3 Compounds Example 2 (Ex. 02) and Example 4 (Ex. 04) were tested on AsPC-1 cells for degradation of KRAS G12D in time-dependent manner and inhibition on ERK1/2 activation; rescue of KRAS G12D degradation is observed with co-incubation with proteasome inhibitor MG132 for 24h; the indicated compounds were applied to AsPC-1 cells at 1 uM for 4h, 8h, 16h, 24h, 48h and 72h; and lysates from these treated cell samples were analyzed for the protein level of KRAS, ERK activation (P-Erk) .
  • P-Erk protein level of KRAS, ERK activation
  • FIG. 4 Selectivity towards KRAS G12D protein by Example 2 and Example 4 compounds.
  • DMSO vehicle
  • an E3 ubiquitin ligase e.g., a Von Hippel-Lindau (VHL) E3 ubiquitin ligase
  • VHL Von Hippel-Lindau
  • the present disclosure provides the compounds and compositions comprising an E3 ubiquitin ligase binding moiety ( “U” or “ULM” ) coupled by a bond or chemical linking group (Linker) to a protein targeting moiety ( “P” ) that targets the KRas protein, which results in the ubiquitination of the KRas protein, and which leads to degradation of the KRas protein by the proteasome.
  • U E3 ubiquitin ligase binding moiety
  • Linker chemical linking group
  • the description provides compounds in which the P binds to the KRas protein and/or a mutated form thereof.
  • the present disclosure also provides a library of compositions and the use thereof to produce targeted degradation of the KRas protein in a cell.
  • the present disclosure provides hetero-bifunctional compounds which comprise a ligand, e.g., a small molecule ligand (i.e., having a molecular weight of below 2,000, 1,000, 500, or 200 Daltons) , which is capable of binding to an E3 ubiquitin ligase, such as the Von Hippel-Lindau E3 ubiquitin ligase.
  • a ligand e.g., a small molecule ligand (i.e., having a molecular weight of below 2,000, 1,000, 500, or 200 Daltons)
  • E3 ubiquitin ligase such as the Von Hippel-Lindau E3 ubiquitin ligase.
  • the compounds also comprise a small molecule moiety that is capable of binding to the KRas protein or mutated form thereof in such a way that the KRas protein or mutated form is placed in proximity to the ubiquitin ligase to effect ubiquitination and degradation (and/or inhibition) of the KRas protein or mutated form.
  • “Small molecule” means, in addition to the above, that the molecule is non-peptidyl, that is, it is not considered a peptide, e.g., comprises fewer than 4, 3, or 2 amino acid residues.
  • each of the P, U and hetero-bifunctional molecule is a small molecule.
  • KRas as used throughout the Specification, unless specifically indicated to the contrary, is intended to include both wild-type KRas and mutant forms therefore, such as a gain-of-function KRas mutant protein or a KRas protein having one or more mutation selected from codon 12 missense mutation, exon 2 mutation, G12V, G12C, G12D, G12A, G13D, exon 3 mutation, codon 61 missense mutation, exon 4 mutation, G12R, Q61H, G12S, A146T, G13C, Q61R, Q61L, A146V, codon 117 missense mutation, KI 17N, Q61K, G12F, codon 59 missense mutation, A59T, or combinations thereof or combinations thereof.
  • compound refers to any specific hetero-bifunctional compound disclosed herein, pharmaceutically acceptable salts and solvates thereof, and deuterated forms of any of the aforementioned molecules, where applicable.
  • Deuterated compounds contemplated are those in which one or more of the hydrogen atoms contained in the drug molecule have been replaced by deuterium. Such deuterated compounds preferably have one or more improved pharmacokinetic or pharmacodynamic properties (e.g., longer half-life) compared to the equivalent “undeuterated” compound.
  • ubiquitin ligase refers to a family of proteins that facilitate the transfer of one or more ubiquitins to a specific substrate protein. Addition of a chain of several ubiquitins (poly-ubiquitination) targets the substrate protein for degradation.
  • Von Hippel-Lindau is an E3 ubiquitin ligase that alone, or in combination with an E2 ubiquitin-conjugating enzyme, can ultimately cause the attachment of a chain of four ubiquitins to a lysine residue on the target protein, thereby targeting the protein for degradation by the proteasome.
  • the ubiquitin ligase is involved in poly-ubiquitination such that a first ubiquitin is attached to a lysine on the target protein; a second ubiquitin is attached to the first; a third is attached to the second, and a fourth is attached to the third.
  • poly-ubiquitination marks proteins for degradation by the proteasome.
  • patient or “subject” is used throughout the specification to describe an animal, preferably a human or a domesticated animal, to whom treatment, including prophylactic treatment, with the compositions according to the present disclosure is provided.
  • a specific animal such as a human patient
  • the term “patient” refers to that specific animal, including a domesticated animal such as a dog or cat, or a farm animal such as a horse, cow, sheep, etc.
  • the terms “patient” and “subject” refer to a human patient unless otherwise stated or implied from the context of the use of the term.
  • alkyl as used herein shall mean within its context a linear, branch-chained or cyclic fully saturated hydrocarbon radical, preferably a C 1 -C 10 , preferably a C 1 -C 6 , or more preferably a C 1 -C 3 alkyl group, which may be optionally substituted with any suitable functional group or groups.
  • alkyl groups are methyl, ethyl, n-butyl, sec-butyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, isopropyl, 2-methylpropyl, cyclopropyl, cyclopropylmethyl, cyclobutyl, cyclopentyl, cyclopentylethyl, cyclohexylethyl and cyclohexyl, among others.
  • the alkyl group is end-capped with a halogen group (Br, Cl, F, or I) .
  • alkynyl refers to linear, branch-chained or cyclic C 2 -C 10 (preferably C 2 -C 6 ) hydrocarbon radicals containing at least one C ⁇ C bond.
  • alkylene when used, refers to a - (CH 2 ) n-group (n is an integer generally from 0-6) , which may be optionally substituted.
  • the alkylene group preferably is substituted on one or more of the methylene groups with a C 1 -C 6 alkyl group (including a cyclopropyl group or a t-butyl group) , but may also be substituted with one or more halo groups, preferably from 1 to 3 halo groups or one or two hydroxyl groups, O- (C 1 -C 6 alkyl) groups or amino acid sidechains as otherwise disclosed herein.
  • an alkylene group may be substituted with a urethane or alkoxy group (or other suitable functional group) which may be further substituted with a polyethylene glycol chain (of from 1 to 10, preferably 1 to 6, or more preferably 1 to 4 ethylene glycol units) to which is substituted (preferably, but not exclusively on the distal end of the polyethylene glycol chain) an alkyl chain substituted with a single halogen group, preferably a chlorine group.
  • the alkylene (e.g., methylene) group may be substituted with an amino acid sidechain group such as a sidechain group of a natural or unnatural amino acid, for example, alanine, ⁇ -alanine, arginine, asparagine, aspartic acid, cysteine, cystine, glutamic acid, glutamine, glycine, phenylalanine, histidine, isoleucine, lysine, leucine, methionine, proline, serine, threonine, valine, tryptophan or tyrosine.
  • an amino acid sidechain group such as a sidechain group of a natural or unnatural amino acid, for example, alanine, ⁇ -alanine, arginine, asparagine, aspartic acid, cysteine, cystine, glutamic acid, glutamine, glycine, phenylalanine, histidine, isoleucine, lysine, leucine, methion
  • unsubstituted shall mean substituted only with hydrogen atoms.
  • a range of carbon atoms which includes C 0 means that carbon is absent and is replaced with H.
  • a range of carbon atoms which is C 0 -C 6 includes carbons atoms of 1, 2, 3, 4, 5 and 6 and for C 0 , H stands in place of carbon.
  • substituted or “optionally substituted” shall mean independently (i.e., where more than one substituent occurs, each substituent is selected independent of another substituent) one or more substituents (independently up to five substituents, preferably up to three substituents, more preferably 1 or 2 substituents on a moiety in a compound according to the present disclosure and may include substituents which themselves may be further substituted) at a carbon (or nitrogen) position anywhere on a molecule within context, and includes as possible substituents such as hydroxyl, thiol, carboxyl, cyano (C ⁇ N) , nitro (NO 2 ) , halogen (preferably, 1, 2 or 3 halogens, especially on an alkyl, especially a methyl group such as a trifluoromethyl) , an alkyl group (preferably, C 1 -C 10 , more preferably, C 1 -C 6 ) , aryl (especially phenyl and substituted phenyl, for example
  • Substituents according to the present disclosure may include, for example –SiR 1 R 2 R 3 groups where each of R 1 and R 2 is as otherwise described herein and R 3 is H or a C 1 -C 6 alkyl group, preferably R 1 , R 2 , R 3 together is a C 1 -C 3 alkyl group (including an isopropyl or t-butyl group) .
  • Each of the above-described groups may be linked directly to the substituted moiety or alternatively, the substituent may be linked to the substituted moiety (preferably in the case of an aryl or heteroaryl moiety) through an optionally substituted - (CH 2 ) m-or alternatively an optionally substituted - (OCH 2 ) m-, - (OCH 2 CH 2 ) m-or - (CH 2 CH 2 O) m-group, which may be substituted with any one or more of the above-described substituents.
  • Alkylene groups - (CH 2 ) m -or - (CH 2 ) n -groups or other chains such as ethylene glycol chains, as identified above, may be substituted anywhere on the chain.
  • Preferred substituents on alkylene groups include halogen or C 1 -C 6 (preferably C 1 -C 3 ) alkyl groups, which may be optionally substituted with one or two hydroxyl groups, one or two ether groups (O-C 1 -C 6 groups) , up to three halo groups (preferably F) , or a side chain of an amino acid as otherwise described herein and optionally substituted amide (preferably carboxamide substituted as described above) or urethane groups (often with one or two C 0 -C 6 alkyl substituents, which group (s) may be further substituted) .
  • halogen or C 1 -C 6 (preferably C 1 -C 3 ) alkyl groups which may be optionally substituted with one or two hydroxyl groups, one or two ether groups (O-C 1 -C 6 groups) , up to three halo groups (preferably F) , or a side chain of an amino acid as otherwise described herein and optionally substituted amide (preferably car
  • the alkylene group (often a single methylene group) is substituted with one or two optionally substituted C 1 -C 6 alkyl groups, preferably C 1 -C 4 alkyl group, most often methyl or O-methyl groups or a sidechain of an amino acid as otherwise described herein.
  • a moiety in a molecule may be optionally substituted with up to five substituents, preferably up to three substituents. Most often, in the present disclosure moieties which are substituted are substituted with one or two substituents.
  • substituted (each substituent being independent of any other substituent) shall also mean within its context of use C 1 -C 6 alkyl, C 1 -C 6 alkoxy, halogen, amido, carboxamido, sulfone, including sulfonamide, keto, carboxy, C 1 -C 6 ester (oxyester or carbonylester) , C 1 -C 6 keto, urethane -O-C (O) -NR 1 R 2 or –N (R 1 ) -C (O) -O-R 1 , nitro, cyano and amine (especially including a C 1 -C 6 alkylene-NR 1 R 2 , a mono-or di-C 1 -C 6 alkyl substituted amines which may be optionally substituted with one or two hydroxyl groups) .
  • R 1 and R 2 are each, within context, H or a C 1 -C 6 alkyl group (which may be optionally substituted with one or two hydroxyl groups or up to three halogen groups, preferably fluorine) .
  • substituted shall also mean, within the chemical context of the compound defined and substituent used, an optionally substituted aryl or heteroaryl group or an optionally substituted heterocyclic group as otherwise described herein.
  • Alkylene groups may also be substituted as otherwise disclosed herein, preferably with optionally substituted C 1 -C 6 alkyl groups (methyl, ethyl or hydroxymethyl or hydroxyethyl is preferred, thus providing a chiral center) , a sidechain of an amino acid group as otherwise described herein, an amido group as described hereinabove, or a urethane group O-C (O) -NR 1 R 2 group where R 1 and R 2 are as otherwise described herein, although numerous other groups may also be used as substituents.
  • Various optionally substituted moieties may be substituted with 3 or more substituents, preferably no more than 3 substituents and preferably with 1 or 2 substituents.
  • aryl or “aromatic” , in context, refers to a substituted (as otherwise described herein) or unsubstituted monovalent aromatic radical (e.g., a 5-16 membered ring) having a single ring (e.g., benzene, phenyl, benzyl, or 5, 6, 7 or 8 membered ring) or condensed rings (e.g., naphthyl, anthracenyl, phenanthrenyl, 10-16 membered ring, etc. ) and can be bound to the compound according to the present disclosure at any available stable position on the ring (s) or as otherwise indicated in the chemical structure presented.
  • monovalent aromatic radical e.g., a 5-16 membered ring
  • condensed rings e.g., naphthyl, anthracenyl, phenanthrenyl, 10-16 membered ring, etc.
  • aryl groups in context, may include heterocyclic aromatic ring systems, “heteroaryl” groups having one or more nitrogen, oxygen, or sulfur atoms in the ring (moncyclic) such as imidazole, furyl, pyrrole, furanyl, thiene, thiazole, pyridine, pyrimidine, pyrazine, triazole, oxazole or fused ring systems such as indole, quinoline, indolizine, azaindolizine, benzofurazan, etc., among others, which may be optionally substituted as described above.
  • heteroaryl groups having one or more nitrogen, oxygen, or sulfur atoms in the ring (moncyclic) such as imidazole, furyl, pyrrole, furanyl, thiene, thiazole, pyridine, pyrimidine, pyrazine, triazole, oxazole or fused ring systems such as indole, quinoline, indolizin
  • heteroaryl groups include nitrogen-containing heteroaryl groups such as pyrrole, pyridine, pyridone, pyridazine, pyrimidine, pyrazine, pyrazole, imidazole, triazole, triazine, tetrazole, indole, isoindole, indolizine, azaindolizine, purine, indazole, quinoline, dihydroquinoline, tetrahydroquinoline, isoquinoline, dihydroisoquinoline, tetrahydroisoquinoline, quinolizine, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, pteridine, imidazopyridine, imidazotriazine, pyrazinopyridazine, acridine, phenanthridine, carbazole, carbazoline, pyrimidine, phenanthroline
  • substituted aryl refers to an aromatic carbocyclic group comprised of at least one aromatic ring or of multiple condensed rings at least one of which being aromatic, wherein the ring (s) are substituted with one or more substituents.
  • an aryl group can comprise a substituent (s) selected from: - (CH 2 ) nOH, - (CH 2 ) n-O- (C 1 -C 6 ) alkyl, - (CH 2 ) n-O- (CH 2 ) n- (C 1 -C 6 ) alkyl, - (CH 2 ) n-C (O) (C 0 -C 6 ) alkyl, - (CH 2 ) n-C (O) O (C 0 -C 6 ) alkyl, - (CH 2 ) n-OC (O) (C 0 -C 6 ) alkyl, amine, mono-or di- (C 1 -C 6 alkyl) amine wherein the alkyl group on the amine is optionally substituted with 1 or 2 hydroxyl groups or up to three halo (preferably F, Cl) groups, OH, COOH, C 1 -C 6 alkyl, preferably CH 3
  • Carboxyl denotes the group --C (O) OR, where R is hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl or substituted heteroaryl, whereas these generic substituents have meanings which are identical with definitions of the corresponding groups defined herein.
  • heteroaryl or “hetaryl” can mean but is in no way limited to a 5-16 membered heteroaryl (e.g., 5, 6, 7 or 8 membered monocylic ring or a 10-16 membered heteroaryl having multiple condensed rings) , an optionally substituted quinoline (which may be attached to the pharmacophore or substituted on any carbon atom within the quinoline ring) , an optionally substituted indole (including dihydroindole) , an optionally substituted indolizine, an optionally substituted azaindolizine (2, 3 or 4-azaindolizine) an optionally substituted benzimidazole, benzodiazole, benzoxofuran, an optionally substituted imidazole, an optionally substituted isoxazole, an optionally substituted oxazole (preferably methyl substituted) , an optionally substituted diazole, an optionally substituted triazole, a
  • S c is CHR SS , NR URE , or O;
  • R HET is H, CN, NO 2 , halo (preferably Cl or F) , optionally substituted C 1 -C 6 alkyl (preferably substituted with one or two hydroxyl groups or up to three halo groups (e.g. CF 3 ) , optionally substituted O (C 1 -C 6 alkyl) (preferably substituted with one or two hydroxyl groups or up to three halo groups) or an optionally substituted acetylenic group –C ⁇ C-Ra where Ra is H or a C 1 -C 6 alkyl group (preferably C 1 -C 3 alkyl) ;
  • R SS is H, CN, NO 2 , halo (preferably F or Cl) , optionally substituted C 1 -C 6 alkyl (preferably substituted with one or two hydroxyl groups or up to three halo groups) , optionally substituted O- (C 1 -C 6 alkyl) (preferably substituted with one or two hydroxyl groups or up to three halo groups) or an optionally substituted -C (O) (C 1 -C 6 alkyl) (preferably substituted with one or two hydroxyl groups or up to three halo groups) ;
  • R URE is H, a C 1 -C 6 alkyl (preferably H or C 1 -C 3 alkyl) or a –C (O) (C 1 -C 6 alkyl) , each of which groups is optionally substituted with one or two hydroxyl groups or up to three halogen, preferably fluorine groups, or an optionally substituted heterocycle, for example piperidine, morpholine, pyrrolidine, tetrahydrofuran, tetrahydrothiophene, piperidine, piperazine, each of which is optionally substituted, and
  • Y C is N or C-R YC , where R YC is H, OH, CN, NO 2 , halo (preferably Cl or F) , optionally substituted C 1 -C 6 alkyl (preferably substituted with one or two hydroxyl groups or up to three halo groups (e.g. CF 3 ) , optionally substituted O (C 1 -C 6 alkyl) (preferably substituted with one or two hydroxyl groups or up to three halo groups) or an optionally substituted acetylenic group –C ⁇ C-Ra where Ra is H or a C 1 -C 6 alkyl group (preferably C 1 -C 3 alkyl) .
  • R YC is H, OH, CN, NO 2 , halo (preferably Cl or F) , optionally substituted C 1 -C 6 alkyl (preferably substituted with one or two hydroxyl groups or up to three halo groups (e.g. CF 3 ) , optionally
  • aralkyl and heteroarylalkyl refer to groups that comprise both aryl or, respectively, heteroaryl as well as alkyl and/or heteroalkyl and/or carbocyclic and/or heterocycloalkyl ring systems according to the above definitions.
  • arylalkyl refers to an aryl group as defined above appended to an alkyl group defined above.
  • the arylalkyl group is attached to the parent moiety through an alkyl group wherein the alkyl group is one to six carbon atoms.
  • the aryl group in the arylalkyl group may be substituted as defined above.
  • Heterocycle refers to a cyclic group which contains at least one heteroatom, e.g., N, O or S, and may be aromatic (heteroaryl) or non-aromatic.
  • heteroaryl moieties are subsumed under the definition of heterocycle, depending on the context of its use. Exemplary heteroaryl groups are described hereinabove.
  • heterocyclics include: azetidinyl, benzimidazolyl, 1, 4-benzodioxanyl, 1, 3-benzodioxolyl, benzoxazolyl, benzothiazolyl, benzothienyl, dihydroimidazolyl, dihydropyranyl, dihydrofuranyl, dioxanyl, dioxolanyl, ethyleneurea, 1, 3-dioxolane, 1, 3-dioxane, 1, 4-dioxane, furyl, homopiperidinyl, imidazolyl, imidazolinyl, imidazolidinyl, indolinyl, indolyl, isoquinolinyl, isothiazolidinyl, isothiazolyl, isoxazolidinyl, isoxazolyl, morpholinyl, naphthyridinyl, oxazolidinyl, oxazolyl, pyr
  • Heterocyclic groups can be optionally substituted with a member selected from the group consisting of alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, keto, thioketo, carboxy, carboxyalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclic, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, -SOary
  • heterocyclic groups can have a single ring or multiple condensed rings.
  • nitrogen heterocycles and heteroaryls include, but are not limited to, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine, naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, phenanthroline, isothiazole, phenazine, isoxazole, phenoxazine, phenothiazine, imidazolidine, imidazoline, piperidine, piperazine, indoline, morpholino, piperidinyl, tetrahydrofur
  • heterocyclic also includes bicyclic groups in which any of the heterocyclic rings is fused to a benzene ring or a cyclohexane ring or another heterocyclic ring (for example, indolyl, quinolyl, isoquinolyl, tetrahydroquinolyl, and the like) .
  • cycloalkyl can mean but is in no way limited to univalent groups derived from monocyclic or polycyclic alkyl groups or cycloalkanes, as defined herein, e.g., saturated monocyclic hydrocarbon groups having from three to twenty carbon atoms in the ring, including, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and the like.
  • substituted cycloalkyl can mean but is in no way limited to a monocyclic or polycyclic alkyl group and being substituted by one or more substituents, for example, amino, halogen, alkyl, substituted alkyl, carbyloxy, carbylmercapto, aryl, nitro, mercapto or sulfo, whereas these generic substituent groups have meanings which are identical with definitions of the corresponding groups as defined in this legend.
  • Heterocycloalkyl refers to a monocyclic or polycyclic alkyl group in which at least one ring carbon atom of its cyclic structure being replaced with a heteroatom selected from the group consisting of N, O, S or P.
  • Substituted heterocycloalkyl refers to a monocyclic or polycyclic alkyl group in which at least one ring carbon atom of its cyclic structure being replaced with a heteroatom selected from the group consisting of N, O, S or P and the group is containing one or more substituents selected from the group consisting of halogen, alkyl, substituted alkyl, carbyloxy, carbylmercapto, aryl, nitro, mercapto or sulfo, whereas these generic substituent group have meanings which are identical with definitions of the corresponding groups as defined in this legend.
  • hydrocarbyl shall mean a compound which contains carbon and hydrogen and which may be fully saturated, partially unsaturated or aromatic and includes aryl groups, alkyl groups, alkenyl groups and alkynyl groups.
  • lower alkyl refers to methyl, ethyl or propyl
  • lower alkoxy refers to methoxy, ethoxy or propoxy.
  • Enantiomers refer to two stereoisomers of a compound which are non-superimposable mirror images of one another. Where the compounds disclosed herein possess two or more asymmetric centers, they may additionally exist as diastereomers. Enantiomers and diastereomers fall within the broader class of stereoisomers. All such possible stereoisomers as substantially pure resolved enantiomers, racemic mixtures thereof, as well as mixtures of diastereomers are intended to be included. All stereoisomers of the compounds disclosed herein and/or pharmaceutically acceptable salts thereof are intended to be included. Unless specifically mentioned otherwise, reference to one isomer applies to any of the possible isomers. Whenever the isomeric composition is unspecified, all possible isomers are included.
  • the term “substantially pure” as used herein means that the target stereoisomer contains no more than 35%, such as no more than 30%, further such as no more than 25%, even further such as no more than 20%, by weight of any other stereoisomer (s) . In some embodiments, the term “substantially pure” means that the target stereoisomer contains no more than 10%, for example, no more than 5%, such as no more than 1%, by weight of any other stereoisomer (s) .
  • substituents found on such ring system may adopt cis and trans formations.
  • Cis formation means that both substituents are found on the upper side of the 2 substituent placements on the carbon, while trans would mean that they were on opposing sides.
  • the di-substituted cyclic ring system may be cyclohexyl or cyclobutyl ring.
  • reaction products from one another and/or from starting materials.
  • the desired products of each step or series of steps is separated and/or purified (hereinafter separated) to the desired degree of homogeneity by the techniques common in the art.
  • separations involve multiphase extraction, crystallization from a solvent or solvent mixture, distillation, sublimation, or chromatography.
  • Chromatography can involve any number of methods including, for example: reverse-phase and normal phase; size exclusion; ion exchange; high, medium and low pressure liquid chromatography methods and apparatus; small scale analytical; simulated moving bed ( "SMB” ) and preparative thin or thick layer chromatography, as well as techniques of small scale thin layer and flash chromatography.
  • SMB simulated moving bed
  • Diastereomers refer to stereoisomers of a compound with two or more chiral centers but which are not mirror images of one another. Diastereomeric mixtures can be separated into their individual diastereomers on the basis of their physical or chemical differences by methods well known to those skilled in the art, such as by chromatography and/or fractional crystallization.
  • Enantiomers can be separated by converting the enantiomeric mixture into a diastereomeric mixture by reaction with an appropriate optically active compound (e.g., chiral auxiliary such as a chiral alcohol or Mosher’s acid chloride) , separating the diastereomers and converting (e.g., hydrolyzing) the individual diastereoisomers to the corresponding pure enantiomers.
  • an appropriate optically active compound e.g., chiral auxiliary such as a chiral alcohol or Mosher’s acid chloride
  • Enantiomers and diastereomers can also be separated by the use of a chiral HPLC column.
  • a single stereoisomer e.g., a substantially pure enantiomer
  • Racemic mixtures of chiral compounds of the invention can be separated and isolated by any suitable method, including: (1) formation of ionic, diastereomeric salts with chiral compounds and separation by fractional crystallization or other methods, (2) formation of diastereomeric compounds with chiral derivatizing reagents, separation of the diastereomers, and conversion to the pure stereoisomers, and (3) separation of the substantially pure or enriched stereoisomers directly under chiral conditions. See: Wainer, Irving W., Ed. Drug Stereochemistry: Analytical Methods and Pharmacology. New York: Marcel Dekker, Inc., 1993.
  • “Pharmaceutically acceptable salts” refer to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio.
  • a pharmaceutically acceptable salt may be prepared in situ during the final isolation and purification of the compounds disclosed herein, or separately by reacting the free base function with a suitable organic acid or by reacting the acidic group with a suitable base.
  • the free base can be obtained by basifying a solution of the acid salt.
  • an addition salt such as a pharmaceutically acceptable addition salt, may be produced by dissolving the free base in a suitable organic solvent and treating the solution with an acid, in accordance with conventional procedures for preparing acid addition salts from base compounds.
  • a pharmaceutically acceptable salt thereof includes salts of at least one compound of Formula (I) , and salts of the stereoisomers of the compound of Formula (I) , such as salts of enantiomers, and/or salts of diastereomers.
  • administering when applied to an animal, human, experimental subject, cell, tissue, organ, or biological fluid, mean contact of an exogenous pharmaceutical, therapeutic, diagnostic agent, or composition to the animal, human, subject, cell, tissue, organ, or biological fluid.
  • Treatment of a cell encompasses contact of a reagent to the cell, as well as contact of a reagent to a fluid, where the fluid is in contact with the cell.
  • administration also means in vitro and ex vivo treatments, e.g., of a cell, by a reagent, diagnostic agent, binding compound, or by another cell.
  • subject herein includes any organism, preferably an animal, more preferably a mammal (e.g., rat, mouse, dog, cat, and rabbit) and most preferably a human.
  • an effective amount refers to an amount of the active ingredient, such as compound that, when administered to a subject for treating a disease, or at least one of the clinical symptoms of a disease or disorder, is sufficient to affect such treatment for the disease, disorder, or symptom.
  • the “therapeutically effective amount” can vary with the compound, the disease, disorder, and/or symptoms of the disease or disorder, severity of the disease, disorder, and/or symptoms of the disease or disorder, the age of the subject to be treated, and/or the weight of the subject to be treated. An appropriate amount in any given instance can be apparent to those skilled in the art or can be determined by routine experiments.
  • “therapeutically effective amount” is an amount of at least one compound and/or at least one stereoisomer thereof, and/or at least one pharmaceutically acceptable salt thereof disclosed herein effective to “treat” as defined herein, a disease or disorder in a subject.
  • the “therapeutically effective amount” refers to the total amount of the combination objects for the effective treatment of a disease, a disorder or a condition.
  • the pharmaceutical composition comprising the compound disclosed herein can be administrated via oral, inhalation, rectal, parenteral or topical route to a subject in need thereof.
  • the pharmaceutical composition may be a regular solid formulation such as tablets, powder, granule, capsules and the like, a liquid formulation such as water or oil suspension or other liquid formulation such as syrup, solution, suspension or the like; for parenteral administration, the pharmaceutical composition may be solution, water solution, oil suspension concentrate, lyophilized powder or the like.
  • the formulation of the pharmaceutical composition is selected from tablet, coated tablet, capsule, suppository, nasal spray or injection, more preferably tablet or capsule.
  • the pharmaceutical composition can be a single unit administration with an accurate dosage.
  • the pharmaceutical composition may further comprise additional active ingredients.
  • compositions disclosed herein can be produced by the conventional methods in the pharmaceutical field.
  • the active ingredient can be mixed with one or more excipients, then to make the desired formulation.
  • the “pharmaceutically acceptable excipient” refers to conventional pharmaceutical carriers suitable for the desired pharmaceutical formulation, for example: a diluent, a vehicle such as water, various organic solvents, etc., a filler such as starch, sucrose, etc., a binder such as cellulose derivatives, alginates, gelatin and polyvinylpyrrolidone (PVP) ; a wetting agent such as glycerol; a disintegrating agent such as agar, calcium carbonate and sodium bicarbonate; an absorption enhancer such as quaternary ammonium compound; a surfactant such as hexadecanol; an absorption carrier such as Kaolin and soap clay; a lubricant such as talc, calcium stearate, magnesium stearate, polyethylene glycol, etc.
  • the pharmaceutical composition further comprises other pharmaceutically acceptable excipients such as a decentralized agent, a stabilizer, a thickener, a complexing agent, a buffering agent, a permeation enhancer, a polymer, an aromatic, a sweetener, a dye and etc.
  • other pharmaceutically acceptable excipients such as a decentralized agent, a stabilizer, a thickener, a complexing agent, a buffering agent, a permeation enhancer, a polymer, an aromatic, a sweetener, a dye and etc.
  • disease refers to any disease, discomfort, illness, symptoms or indications, and can be interchangeable with the term “disorder” or “condition” .
  • C n-m indicates a range which includes the endpoints, wherein n and m are integers and indicate the number of carbons. Examples include C 1-8 , C 1-6 , and the like.
  • the description provides the compounds comprising an E3 ubiquitin ligase binding moiety ( “U” ) that is a VHL E3 ubiquitin ligase binding moiety (a “VLM” ) ,
  • the VLM is covalently coupled to a protein targeting moiety (P) that binds to the protein, which coupling is either directly by a bond or via a chemical linking group (L) according to the structure:
  • L is the bond or chemical linking group
  • P is a protein targeting moiety that binds to the protein KRas or a mutant form thereof, as described herein, where the P is a KRas targeting moiety.
  • VLM is inclusive of all VHL binding moieties.
  • the P is represented by the chemical structure (I) :
  • X is independently selected from C and N,
  • R 1 , R 2 and R 3 are independently selected from H, C 1-6 alkyl, halogen (e.g., F, Cl, Br) , C 1-6 haloalkyl, C 1-6 alkoxy and optionally substituted amino, provided that when X is N, R 1 , R 2 or R 3 attached to X are absent, preferably, R 1 , R 2 and R 3 are independently selected from H, C 1-3 alkyl, halogen (e.g., F, Cl, Br) , more preferably, R 1 , R 2 and R 3 are independently absent or F;
  • halogen e.g., F, Cl, Br
  • R 4 is independently selected from H, C 1-6 alkyl, halogen (e.g., F, Cl, Br) , C 1-6 alkoxy, C 1-6 haloalkyl, optionally substituted amino, C 2-6 alkenyl, C 2-6 alkynyl and C 3-6 cycloalkyl, preferably, R 4 is independently selected from H, C 1-6 alkyl, halogen (e.g., F, Cl, Br) , more preferably, R 4 is independently selected from H, C 1-3 alkyl, F and Cl,
  • R 5 is independently selected from H, C 1-6 alkyl, halogen (e.g., F, Cl, Br) , C 1-6 alkoxy, optionally substituted amino, C 2-6 alkenyl, C 2-6 alkynyl and C 3-6 cycloalkyl, preferably, R 5 is independently selected from H, C 1-3 alkyl, halogen (e.g., F, Cl, Br) , more preferably, R 3 is independently H,
  • connection site denotes a connection site.
  • the P is represented by the following chemical structures (I-1 to I-4) :
  • R 1 , R 2 and R 3 are independently selected from H, C 1-6 alkyl, halogen (e.g., F, Cl, Br) , and C 1-6 haloalkyl;
  • R 4 is independently selected from H, C 1-6 alkyl, and halogen (e.g., F, Cl, Br) ,
  • R 5 is independently selected from H, C 1-3 alkyl, and halogen (e.g., F, Cl, Br) , and
  • connection site denotes a connection site.
  • R 1 , R 2 and R 3 are independently selected from H, C 1-3 alkyl, halogen (e.g., F, Cl, Br) , C 1-6 haloalkyl; preferably, R 1 is F and R 2 and R 3 are H,
  • R 5 is independently selected from H, C 1-3 alkyl, halogen (e.g., F, Cl, Br) , preferably R 5 is H, and
  • connection site denotes a connection site.
  • R 1 and R 3 are independently selected from H, C 1-3 alkyl, halogen (e.g., F, Cl, Br) , C 1-3 haloalkyl; preferably, R 1 is F and R 3 are H,
  • R 5 is independently selected from H, C 1-3 alkyl, halogen (e.g., F, Cl, Br) , preferably, R 5 is H, and
  • connection site denotes a connection site.
  • R 1 and R 2 are independently selected from H, C 1-3 alkyl, halogen (e.g., F, Cl, Br) , C 1-3 haloalkyl; preferably, R 1 is F and R 2 are H,
  • R 5 is independently selected from H, C 1-3 alkyl, halogen (e.g., F, Cl, Br) , preferably, R 5 is H, and
  • connection site denotes a connection site.
  • R 2 and R 3 are independently selected from H, C 1-3 alkyl, halogen (e.g., F, Cl, Br) , C 1-3 haloalkyl; preferably, R 2 is F and R 3 are H,
  • R 5 is independently selected from H, C 1-3 alkyl, halogen (e.g., F, Cl, Br) , preferably, R 5 is H, and
  • connection site denotes a connection site.
  • the E3 ubiquitin ligase binding moiety ( “U” ) is a VLM and is represented by the following chemical structure:
  • R 14 is H or a linear or branched C1-C3 alkyl (e.g., methyl) ;
  • R 15 is a CN or a 5-membered heteroaryl having one or two heteroatoms selected from N, S and O, optionally substituted with a methyl (e.g., ) ;
  • R 16 is a halo, optionally substituted C1-C3 alkyl, optionally substituted C1-C3 haloalkyl, hydroxyl, optionally substituted C1-C3 alkoxy, or optionally substituted C1-C3 haloalkoxy;
  • o is an integer from 0-2 (e.g., 0, 1 or 2) ;
  • U is VLM and comprises a chemical structure selected from the group U-a:
  • R Y1 , R Y2 of Formula U-a are each independently selected from the group of H, linear or branched C 1-6 alkyl, optionally substituted by 1 or more halo, optionally substituted C 1-6 alkoxyl (e.g., optionally substituted by 0-3 R P groups) ;
  • W 3 of Formula U-a is selected from the group of an optionally substituted T, an optionally substituted -T-N (R 1a R 1b ) X 3 , optionally substituted -T-N (R 1a R 1b ) , optionally substituted -T-Aryl, an optionally substituted -T-Heteroaryl, an optionally substituted T-biheteroaryl, an optionally substituted -T-Heterocycle, an optionally substituted -T-biheterocycle, an optionally substituted -NR 1 -T-Aryl, an optionally substituted -NR 1 -T-Heteroaryl, or an optionally substituted -NR 1 -T-Heterocycle;
  • T of Formula U-a is selected from the group of an optionally substituted alkyl, - (CH 2 ) n-group, – (CH 2 ) n-O-C 1 -C 6 alkyl which is optionally substituted, linear, branched, or - (CH 2 ) n-O-heterocyclyl which is optionally substituted, wherein each one of the methylene groups is optionally substituted with one or two substituents selected from the group of halogen, methyl, optionally substituted alkoxy, a linear or branched C 1 -C 6 alkyl group optionally substituted by 1 or more halogen, C (O) NR 1 R 1a , or NR 1 R 1a or R 1 and R 1a are joined to form an optionally substituted heterocycle, or -OH groups or an amino acid side chain optionally substituted;
  • W 4 of Formula U-a is an optionally substituted -NR 1 -T-Aryl wherein the aryl group may be optionally substituted with an optionally substituted 5-6 membered heteroaryl or an optionally substituted aryl, an optionally substituted -NR 1 -T-Heteroaryl group with an optionally substituted aryl or an optionally substituted heteroaryl, or an optionally substituted -NR 1 -T-Heterocycle, where -NR 1 is covalently bonded to X 2 and R 1 is H or CH 3 , preferably H;
  • n is 0 to 6, often 0, 1, 2, or 3, preferably 0 or 1;
  • T is selected from the group of an optionally substituted alkyl, – (CH 2 ) n-group, wherein each one of the methylene groups is optionally substituted with one or two substituents selected from the group of halogen, methyl, optionally substituted alkoxy, a linear or branched C 1 -C 6 alkyl group optionally substituted by 1 or more halogen, C (O) NR 1 R 1a , or NR 1 R 1a or R 1 and R 1a are joined to form an optionally substituted heterocycle, or -OH groups or an amino acid side chain optionally substituted; and n is 0 to 6, often 0, 1, 2, or 3, preferably 0 or 1.
  • W 4 is selected from
  • W 5 is selected from the group of an optionally substituted phenyl, an optionally substituted napthyl, or an optionally substituted 5-10 membered heteroaryl (e.g., W 5 is optionally substituted with one or more [such as 1, 2, 3, 4, or 5] halo, CN, optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted alkoxy, hydroxy, or optionally substituted haloalkoxy) ,
  • R 15 is selected from the group of H, halogen, CN, OH, NO 2 , NR 14a R 14b , OR 14a , CONR 14a R 14b , NR 14a COR 14b , SO 2 NR 14a R 14b , NR 14a SO 2 R 14b , optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted haloalkoxy; optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, or optionally substituted cycloheteroalkyl,
  • R 14a , R 14b are each independently selected from the group of H, haloalkyl (e.g., fluoalkyl) , optionally substituted alkyl, optionally substituted alkoxy, optionally substituted hydroxyl alkyl, optionally substituted alkylamine, optionally substituted heterolkyl, optionally substituted alkyl-heterocycloalkyl, optionally substituted alkoxy-heterocycloalkyl, COR 26 , CONR 27a R 27b , NHCOR 26 , or NHCH 3 COR 26 ; and the other of R 14a and R 14b is H; or R 14a , R 14b , together with the carbon atom to which they are attached, form an optionally substituted 3 to 5 membered cycloalkyl, heterocycloalkyl, spirocycloalkyl or spiroheterocyclyl, wherein the spiroheterocyclyl is not epoxide or azi
  • o is an integer from 0-4 (e.g., 0, 1, 2, 3, or 4) ;
  • R 16 is independently selected from the group of halo, optionally substituted alkyl, optionally substituted haloalkyl, hydroxy, or optionally substituted haloalkoxy.
  • W 4 substituents for use in the present disclosure also include specifically (and without limitation to the specific compound disclosed) the W 4 substituents which are found in the identified compounds disclosed herein. Each of these W 4 substituents may be used in conjunction with any number of W 3 substituents which are also disclosed herein.
  • U-a is optionally substituted by 0-3 R P groups in the pyrrolidine moiety.
  • the W 3 , W 4 can independently be covalently coupled to a linker which is attached one or more P groups.
  • U is VHL and is represented by the structure:
  • W 3 of Formula U-b is selected from the group of an optionally substituted aryl, optionally substituted heteroaryl, or
  • R 9 and R 10 of Formula U-b are independently hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted hydroxyalkyl, optionally substituted heteroaryl, or haloalkyl, or R 9 , R 10 , and the carbon atom to which they are attached form an optionally substituted cycloalkyl;
  • R 11 of Formula U-b is selected from the group of an optionally substituted heterocyclyl, optionally substituted alkoxy, optionally substituted heteroaryl, optionally substituted aryl,
  • R 12 of Formula U-b is selected from the group of H or optionally substituted alkyl
  • R 13 of Formula U-b is selected from the group of H, optionally substituted alkyl, optionally substituted alkylcarbonyl, optionally substituted (cycloalkyl) alkylcarbonyl, optionally substituted aralkylcarbonyl, optionally substituted arylcarbonyl, optionally substituted (heterocyclyl) carbonyl, or optionally substituted aralkyl;
  • R 14a , R 14b of Formula U-b are each independently selected from the group of H, haloalkyl (e.g. fluoroalkyl) , optionally substituted alkyl, optionally substitute alkoxy, aminomethyl, alkylaminomethyl, alkoxymethyl, optionally substituted hydroxyl alkyl, optionally substituted alkylamine, optionally substituted heterolkyl, optionally substituted alkyl-heterocycloalkyl, optionally substituted alkoxy-heterocycloalkyl, CONR 27a R 27b , CH 2 NHCOR 26 , or (CH 2 ) N (CH 3 ) COR 26 ; and the other of R 14a and R 14b is H; or R 14a , R 14b , together with the carbon atom to which they are attached, form an optionally substituted 3 to 6 membered cycloalkyl, heterocycloalky, spirocycloalkyl or spiroheterocyclyl, where
  • W 5 of Formula U-b is selected from the group of an optionally substituted phenyl or an optionally substituted 5-10 membered heteroaryl (e.g., W 5 is optionally substituted with one or more [such as 1, 2, 3, 4, or 5] halo, CN, optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted alkoxy, hydroxy, or optionally substituted haloalkoxy) ,
  • R 15 of Formula U-b is selected from the group of H, halogen, CN, OH, NO 2 , NR 14a R 14b , OR 14a , CONR 14a R 14b , NR 14a COR 14b , SO 2 NR 14a R 14b , NR 14a SO 2 R 14b , optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted haloalkoxy, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, or optionally substituted cycloheteroalkyl;
  • each R 16 of Formula U-b is independently selected from the group of H, CN, halo, optionally substituted alkyl, optionally substituted haloalkyl, hydroxy, or optionally substituted haloalkoxy;
  • o of Formula U-b is 0, 1, 2, 3, or 4;
  • R 18 of Formula U-b is independently selected from the group of H, halo, optionally substituted alkoxy, cyano, optionally substituted alkyl, haloalkyl, haloalkoxy or a linker;
  • p of Formula U-b is 0, 1, 2, 3, or 4, and
  • R 15 is selected from the group of H, halogen, CN, OH, NO 2 , NR 27a R 27b , OR 27a , CONR 27a R 27b , NR 27a COR 27b , SO 2 NR 27a R 27b , NR 27a SO 2 R 27b , optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted haloalkoxy, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, or optionally substituted heterocyclyl, wherein each R 26 is independently selected from H, optionally substituted alkyl or NR 27a R 27b ; and each R 27a and R 27b is independently H, optionally substituted alkyl, or R 27a and R 27b together with the nitrogen atom to which they are attached form a 4-6 membered heterocyclyl.
  • R 15 is wherein R 17 is H, halo, optionally substituted C 3-6 cycloalkyl, optionally substituted C 1-6 alkyl, optionally substituted C 1-6 alkenyl, and C 1-6 haloalkyl; and X a is S or O.
  • R 17 is selected from the group methyl, ethyl, isopropyl, and cyclopropyl.
  • R 15 is selected from the group consisting of:
  • R 11 is selected from the group consisting of:
  • R 14a and R 14b are each independently selected from the group of H, optionally substituted haloalkyl, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted hydroxyl alkyl, optionally substituted alkylamine, optionally substituted heteroalkyl, optionally substituted alkyl-heterocycloalkyl, optionally substituted alkoxy-heterocycloalkyl, CH 2 OR 30 , CH 2 NHR 30 , CH 2 NCH 3 R 30 , CONR 27a R 27b , CH 2 CONR 27a R 27b , CH 2 NHCOR 26 , or CH 2 NCH 3 COR 26 ; and the other of R 14a and R 14b is H; or R 14a , R 14b , together with the carbon atom to which they are attached, form an optionally substituted 3-to 6-membered cycloalkyl, heterocycloalkyl, spirocycloalkyl or spirohe
  • R 30 is selected from H, alkyl, alkynylalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, arylalkyl or heteroarylalkyl further optionally substituted; R 26 and R 27 are as described above.
  • R 15 is selected from H, halogen, CN, OH, NO 2 , NR 27a R 27b , OR 27a , CONR 27a R 27b , NR 27a COR 27b , SO 2 NR 27a R 27b , NR 27a SO 2 R 27b , optionally substituted alkyl, optionally substituted haloalkyl (e.g.
  • optionally substituted fluoroalkyl optionally substituted haloalkoxy, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, or optionally substituted heterocyclyl, wherein optional substitution of said aryl, heteroaryl, cycloalkyl and heterocycloalkyl includes CH 2 OR 30 , CH 2 NHR 30 , CH 2 NCH 3 R 30 , CONR 27a R 27b , CH 2 CONR 27a R 27b , CH 2 NHCOR 26 or wherein R 26 , R 27 , R 30 and R 14a are as described above.
  • R 14a and R 14b are each independently selected from the group of H, optionally substituted haloalkyl, optionally substituted alkyl, CH 2 OR 30 , CH 2 NHR 30 , CH 2 NCH 3 R 30 , CONR 27a R 27b , CH 2 CONR 27a R 27b , CH 2 NHCOR 26 , or CH 2 NCH 3 COR 26 ; and the other of R 14a and R 14b is H; or R 14a , R 14b , together with the carbon atom to which they are attached, form an optionally substituted 3-to 6-membered spirocycloalkyl or spiroheterocyclyl, wherein the spiroheterocyclyl is not epoxide or aziridine, said spirocycloalkyl or spiroheterocycloalkyl itself being optionally substituted with an alkyl, a haloalkyl, or -COR 33 where R 33 is
  • R 15 is selected from H, halogen, CN, OH, NO 2 , NR 27a R 27b, OR 27a , CONR 27a R 27b , NR 27a COR 27b , SO 2 NR 27a R 27b , NR 27a SO 2 R 27b , optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted haloalkoxy, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted cycloalkyl, or optionally substituted heterocyclyl wherein optional substitution of said aryl, heteroaryl, cycloalkyl and heterocycloalkyl includes CH 2 OR 30 , CH 2 NHR 30 , CH 2 NCH 3 R 30 , CONR 27a R 27b , CH 2 CONR 27a R 27b , CH 2 NHCOR 26 , CH 2 NCH 3 COR 26 or wherein R 26 , R 27 , R 30 and R 14a are asdescribed above.
  • U has a chemical structure selected from the group of:
  • R 1 of Formulas U-c, U-d, and U-e is H, ethyl, isopropyl, tert-butyl, sec-butyl, cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl; optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted hydroxyalkyl, optionally substituted heteroaryl, or haloalkyl;
  • R 14a of Formulas U-c, U-d, and U-e is H, haloalkyl, optionally substituted alkyl, methyl, fluoromethyl, hydroxymethyl, ethyl, isopropyl, or cyclopropyl;
  • R 15 of Formulas U-c, U-d, and U-e is selected from the group consisting of H, halogen, CN, OH, NO 2 , optionally substituted heteroaryl, optionally substituted aryl; optionally substituted alkyl, optionally substituted haloalkyl, optionally substituted haloalkoxy, optionally substituted cycloalkyl, or optionally substituted cycloheteroalkyl;
  • R 3 of U-e is absent or an optionally substituted 5 or 6 membered heteroaryl; and the indicates the site of attachment of at least one P, another U (U’) or a chemical linker moiety coupling at least one P or a U’ or both to U.
  • U comprises a group according to the chemical structure:
  • R 14a of Formula U-f is H, haloalkyl, optionally substituted alkyl, methyl, fluoromethyl, hydroxymethyl, ethyl, isopropyl, or cyclopropyl;
  • R 9 of Formula U-f is H
  • R 10 of Formula U-f is H, ethyl, isopropyl, tert-butyl, sec-butyl, cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl;
  • R 11 of Formula U-f is or optionally substituted heteroaryl
  • p of Formula U-f is 0, 1, 2, 3, or 4;
  • each R 18 of Formula U-f is independently halo, optionally substituted alkoxy, cyano, optionally substituted alkyl, haloalkyl, haloalkoxy or a linker;
  • R 13 of Formula U-f is H, optionally substituted alkyl, optionally substituted alkylcarbonyl, optionally substituted (cycloalkyl) alkylcarbonyl, optionally substituted aralkylcarbonyl, optionally substituted arylcarbonyl, optionally substituted (heterocyclyl) carbonyl, or optionally substituted aralkyl,
  • R 15 of Formula U-f is selected from the group consisting of H, halogen, Cl, CN, OH, NO 2 , optionally substituted haloalkyl, optionally substituted heteroaryl, optionally substituted aryl;
  • the of Formula U-f indicates the site of attachment of at least one P, another U (U’) or a chemical linker moiety coupling at least one P or a U’ or both to U.
  • the VLM is covalently joined to a P, or a chemical linker group (L) via an R group (such as, R P , R 1 , R 1a , R 1b , R Y1 , R Y2 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14a , R 14b , R 15 , R 16 , R 17 , R 18 , R 26 , R 27a , R 27b , R 30 , R 33 ) , W 3 , W 4 , W 5 , X, X 1 , X 2 , X 3 , or T.
  • R group such as, R P , R 1 , R 1a , R 1b , R Y1 , R Y2 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14a , R 14b , R 15 , R 16 , R 17 , R 18 , R 26 , R 27a
  • the VLM is covalently joined to a P, or a chemical linker group (L) via R P , R 1 , R 1a , R 1b , R Y1 , R Y2 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14a , R 14b , R 15 , R 16 , R 17 , R 18 , R 26 , R 27a , R 27b , R 30 , R 33 , W 3 , W 4 , W 5 , X, X 1 , X 2 , X 3 , or T.
  • L chemical linker group
  • the R P , R 1 , R 1a , R 1b , R Y1 , R Y2 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14a , R 14b , R 15 , R 16 , R 17 , R 18 , R 26 , R 27a , R 27b , R 30 , R 33 , W 3 , W 4 , W 5 , X, X 1 , X 2 , X 3 , or T can independently be covalently coupled to a linker and/or a linker to which is attached to one or more P, U, and VLM group.
  • the U is selected from the following structures:
  • the U is selected from the following structures:
  • n is 0 or 1 and the indicates the site of attachment of at least one P, another U (U’) or a chemical linker moiety coupling at least one P or a U’ or both to U.
  • the U is selected from the following structures:
  • the phenyl ring in ULM-a1 through ULM -a15, ULM -b1 through ULM-b12, ULM-c1 through ULM-c15 and ULM-d1 through ULM-d9 is optionally substituted with fluorine, lower alkyl and alkoxy groups, and wherein the indicates the site of attachment of at least one P, another ULM (ULM’) or a chemical linker moiety coupling at least one P or a ULM’ or both to ULM-a.
  • the phenyl ring in ULM-a1 through ULM-a15, ULM-b1 through ULM-b12, ULM-c1 through ULM-c15 and ULM-d1 through ULM-d9 can be functionalized as the ester to make it a part of the prodrug.
  • the hydroxyl group on the pyrrolidine ring of ULM-a1 through ULM-a15, ULM-b1 through ULM-b12, ULM-c1 through ULM-c15 and ULM-d1 through ULM-d9, respectively, comprises an ester-linked prodrug moiety.
  • the U is according to the following formula: or a pharmaceutically acceptable salt thereof,
  • R 1 , R 14a and R 14b are as described herein;
  • X is CH or N
  • R 30 is H, F or Cl
  • R 16 is H, C 1-4 alkyl, fluoro, chloro, CN, or C 1-4 alkoxy;
  • R 28 is H, methyl, CH 2 N (Me) 2 , CH 2 OH, CH 2 O (C 1-4 alkyl) , CH 2 NHC (O) C 1-4 alkyl, NH 2 , and
  • the U is according to the following formula:
  • R 1 , R 14a and R 14b are as described herein;
  • R 30 is H, F or Cl
  • the U (or when present, U’) as described herein may be a pharmaceutically acceptable salt, enantiomer, diastereomer, solvate or polymorph thereof.
  • the U (or when present, U’) as described herein may be coupled to a P directly via a bond or by a chemical linker.
  • the compounds as described herein include a P chemically linked to a U (e.g., VLM) via a chemical linker (L) .
  • the linker group L comprises one or more covalently connected structural units (e.g., -A L 1 ... (A L ) q-or - (A L ) q-) , wherein A L 1 is a group coupled to P, and (A L ) q is a group coupled to U.
  • the linker (L) to a U (e.g., VLM) connection is a stable L-U connection.
  • a linker (L) and a U are connected via a heteroatom (e.g., N, O, S)
  • any additional heteroatom if present, is separated by at least a carbon atom (e.g., -CH 2 -) , such as with an acetal or aminal group.
  • the heteroatom is not part of an ester.
  • the linker group L is a bond or a chemical linker group represented by the formula - (A L ) q-, wherein A is a chemical moiety and q is an integer from 1-100 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, or 80) , and wherein L is covalently bound to both the P and the U, and provides for binding of the P to the protein target and the U to an E3 ubiquitin ligas
  • the linker group L is a bond or a chemical linker group represented by the formula - (A L ) q-, wherein A is a chemical moiety and q is an integer from 1-30 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) , and wherein L is covalently bound to both the P and the U, and provides for binding of the P to the protein target and the U to an E3 ubiquitin ligase in sufficient proximity to result in target protein ubiquitination.
  • the linker group L is - (A L ) q-, wherein: (A L ) q is a group which connects a U (e.g., VLM) , to P (KTM) ;
  • q of the linker is an integer greater than or equal to 1;
  • q is an integer greater than or equal to 1.
  • (A L ) q is a group which is A L 1 and (A L ) q wherein the linker couples a P to a U.
  • a L 2 is a group which is connected to A L 1 and to a U.
  • the structure of the linker group L is -A L 1-, and A L 1 is a group which connects a Umoiety to a P moiety.
  • the unit A L of linker (L) comprises a group represented by a general structure selected from the group consisting of:
  • n of the linker can be 0 to 10;
  • R of the linker can be H, or lower alkyl
  • R1 and R2 of the linker can form a ring with the connecting N.
  • the linker (L) includes an optionally substituted C 1 -C 50 alkyl (e.g., C 1 , C 2 , C 3 , C 4 , C 5 , C 6 , C 7 , C 8 , C 9 , C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C 16 , C 17 , C 18 , C 19 , C 20 , C 21 , C 22 , C 23 , C 24 , C 25 , C 26 , C 27 , C 28 , C 29 , C 30 , C 31 , C 32 , C 33 , C 34 , C 35 , C 36 , C 37 , C 38 , C 39 , C 40 , C 41 , C 42 , C 43 , C 44 , C 45 , C 46 , C 47 , C 48 , C 49 , or C 50 alkyl, and including all implied subranges, e.g., C 1 , C 2 , C 3
  • each carbon is optionally independently substituted or replaced with (1) a heteroatom selected from N, O, S, P, or Si atoms that has an appropriate number of hydrogens, substitutions, or both to complete valency, (2) an optionally substituted cycloalkyl or bicyclic cycloalkly, (3) an optionally substituted heterocyloalkyl or bicyclic heterocyloalkyl, (4) an optionally substituted aryl or bicyclic aryl, or (5) optionally substituted heteroaryl or bicyclic heteroaryl.
  • the linker (L) does not have heteroatom-heteroatom bonding (e.g., no heteroatoms are covalently linked or adjacently located) .
  • the linker (L) includes about 1 to about 50 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50) alkylene glycol units that are optionally substituted, wherein carbon or oxygen may be substituted or replaced with a heteroatom selected from N, S, P, or Si atoms with an appropriate number of hydrogens to complete valency.
  • a heteroatom selected from N, S, P, or Si atoms with an appropriate number of hydrogens to complete valency.
  • linker (L) is represented by the chemical structure: wherein:
  • the of the chemical linking moiety is the site of attachment to the VLM or the P;
  • Y L2 is a bond, or a unsubstituted or substituted linear or branched C1-C4 alkyl (e.g., optionally substituted with a halogen, C1-3 alkyl, methyl, or ethyl) ;
  • W L3 is a 3-7 membered ring (e.g., 4-6 membered cycloalkyl or heterocycloalkyl) or a 8-12 member spirocyclic, each with 0-4 heteroatoms (e.g., 0-4 heteroatoms independently selected from N, O, and S) and optionally substituted with halogen or methyl;
  • W L4 is a 3-8 membered ring (e.g., 4-6 membered cycloalkyl or heterocycloalkyl, or ) or a 5-8 member spirocyclic, each with 0-4 heteroatoms (e.g., 0-4 heteroatoms independently selected from N, O, and S) and optionally substituted with halogen (e.g., F, Cl, Br) , or methyl; and
  • Y L5 is a bond or an unsubstituted or substituted C1-C6 alkyl, where one or more C atoms are optionally replaced with O and optionally substituted with a halo (e.g., F, Cl, Br) , or methyl.
  • a halo e.g., F, Cl, Br
  • the unit A L of the linker (L) comprises a structure selected from the group consisting of:
  • the unit A L of the linker (L) comprises a structure selected from the group consisting of: wherein the indicates the point of attachment with the P or the VLM.
  • the unit A L of the linker (L) comprises a structure selected from the group consisting of:
  • hetero-bifunctional compound is represented by the chemical structure:
  • X is independently selected from C and N,
  • R 1 , R 2 and R 3 are independently selected from H, C 1-6 alkyl, halogen (e.g., F, Cl, Br) , C 1-6 haloalkyl, C 1-6 alkoxy and optionally substituted amino, provided that when X is N, R 1 , R 2 or R 3 attached to X are absent, preferably, R 1 , R 2 and R 3 are independently selected from H, C 1-3 alkyl, halogen (e.g., F, Cl, Br) , more preferably, R 1 , R 2 and R 3 are independently absent or F;
  • halogen e.g., F, Cl, Br
  • R 4 is independently selected from H, C 1-6 alkyl, halogen (e.g., F, Cl, Br) , C 1-6 alkoxy, C 1-6 haloalkyl, optionally substituted amino, C 2-6 alkenyl, C 2-6 alkynyl and C 3-6 cycloalkyl, preferably, R 4 is independently selected from H, C 1-6 alkyl, halogen (e.g., F, Cl, Br) , more preferably, R 4 is independently selected from H, C 1-3 alkyl, F and Cl,
  • R 5 is independently selected from H, C 1-6 alkyl, halogen (e.g., F, Cl, Br) , C 1-6 alkoxy, optionally substituted amino, C 2-6 alkenyl, C 2-6 alkynyl and C 3-6 cycloalkyl, preferably, R 5 is independently selected from H, C 1-3 alkyl, halogen (e.g., F, Cl, Br) , more preferably, R 3 is independently H,
  • connection site denotes a connection site
  • L and VLM are as defined in any aspect or embodiment described herein.
  • hetero-bifunctional compound is represented by the chemical structure:
  • Linker is selected from wherein the indicates the point of attachment.
  • hetero-bifunctional compound is represented by the chemical structure:
  • Linker is selected from
  • hetero-bifunctional compound is represented by the chemical structure:
  • a pharmaceutical composition which comprises at least one bifunctional compound as described herein, a pharmaceutically acceptable salt thereof or stereoisomer thereof and one or more pharmaceutically acceptable excipients.
  • the description provides a method of ubiquitinating/degrading a target protein in a cell.
  • the method comprises administering a bifunctional compound of the invention.
  • the control or reduction of specific protein levels in cells of a subject as afforded by the present disclosure provides treatment of a disease state, condition, or symptom.
  • the method comprises administering an effective amount of a compound as described herein, optionally including a pharmaceutically acceptable excipient, carrier, adjuvant, another bioactive agent or combination thereof.
  • the subject compounds and pharmaceutically acceptable salts thereof can be prepared from (a) commercially available starting materials (b) known starting materials which may be prepared as described in literature procedures (c) new intermediates described in the schemes and experimental procedures herein.
  • the order of synthetic steps may be varied to increase the yield of desired product.
  • reaction flasks were fitted with rubber septa for the introduction of substrates and reagents via syringe; and glassware was oven dried and/or heat dried.
  • Step 1 To a solution of (S) -1- (4-bromophenyl) ethan-1-amine (90 g, 0.45 mol) in DCM (900 mL) was added Boc 2 O (117 g, 0.54 mol) and TEA (90 g, 0.90 mol) . The reaction mixture was stirred at 25 °C for 16 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel to afford tert-butyl (S) - (1- (4-bromophenyl) ethyl) carbamate (110 g) as a white solid.
  • Step 2 A mixture of tert-butyl (S) - (1- (4-bromophenyl) ethyl) carbamate (110 g, 0.37 mol) , 4-methylthiazole (72.8 g, 0.74 mol) , palladium (II) acetate (8.2 g, 0.037 mol) and potassium acetate (72.1 g, 0.74 mol) in DMF (1000 mL) was stirred at 120 °Cunder N 2 for 6 h. After cooling to ambient temperature, the reaction mixture was filtered. To the filtrate was added H 2 O (500 mL) . The aqueous phase was extracted with EtOAc (400 mL x 3) .
  • Step 3 To a solution of tert-butyl (S) - (1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamate (71 g, 0.22 mol) in MeOH (350 mL) was added HCl/1, 4-dioxane (4 M, 350 mL) . The reaction mixture was stirred at 25 °C for 16 h. The reaction mixture was concentrated under reduced pressure to afford (S) -1- (4- (4-Methylthiazol-5-yl) phenyl) ethan-1-amine (58 g) as a yellow solid. LCMS (ESI) m/z: 219.1 [M+1] + .
  • Step 4 A solution of (2S, 4R) -1- (tert-butoxycarbonyl) -4-hydroxypyrrolidine-2-carboxylic acid (61.4 g, 0.26 mol) , DIPEA (102 g, 0.79 mol) and HATU (151 g, 0.39 mol) in DMF (300 mL) was stirred at 25 °C for 15 min. Then (S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethan-1-amine (58 g, 0.26 mol) in DMF (100 mL) was added to the mixture. The reaction mixture was stirred at 25 °C for 16 h. The reaction mixture was poured into water (1000 mL) .
  • Step 5 A solution of tert-butyl (2S, 4R) -4-hydroxy-2- ( ( (S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrroli dine-1-carboxylate (52 g, 0.12 mol) in HCl/1, 4-dioxane (4 M, 300 mL) was stirred at 25 °C for 2 h. The reaction mixture was concentrated under reduced pressure. The residue was triturated with EtOAc (500 mL) to afford Intermediate 1 (40 g) as a yellow solid. LCMS (ESI) m/z: 332.1 [M+1] + .
  • Step 1 A solution of 5-methylisoxazol-3-ol (20 g, 0.20 mol) and K 2 CO 3 (55.7 g, 0.40 mol) in acetone (360 mL) was stirred at 60 °C for 1 h. Then BnBr (41.4 g, 0.24 mol) was added to the mixture at 60 °C. The resulting solution was stirred at 60 °C for 1 h. The mixture was filtered and washed with EtOAc (300 mL) . The combined filtrate was concentrated. The residue was purified with column on silica gel to afford 3- (benzyloxy) -5-methylisoxazole (5.9 g) as a yellow oil. LCMS (ESI) m/z: 190.1 [M+1] + .
  • Step 2 To a solution of LDA (13 mL, 0.026 mol, 2 M in THF) in THF (33 mL) was added a solution of 3- (benzyloxy) -5-methylisoxazole (3.3 g, 0.017 mol) in THF (33 mL) dropwise at -78 °C. After stirring at -78 °C for 1 h, the reaction mixture was poured into solid CO 2 /THF mixture. The mixture was stirred at room temperature for 1 h. Then the pH was adjusted to 1 ⁇ 2 with aq. HCl (6 M) at 0 °C. The mixture was extracted with ethyl acetate (200 mL x 2) .
  • Step 3 To a solution of 2- (3- (benzyloxy) isoxazol-5-yl) acetic acid (2.0 g, 8.6 mmol) in MeOH (25 mL) was added SOCl 2 (3.0 g, 25.7 mmol) dropwise at 0 °C. The resulting solution was stirred at 15 °C for 12 h. The mixture was concentrated under reduced pressure to afford a yellow oil. Then the yellow oil was diluted with water (50 mL) and extracted with EtOAc (50 mL) . The organic phase was concentrated.
  • Step 4 To a solution of methyl 2- (3- (benzyloxy) isoxazol-5-yl) acetate (600 mg, 2.42 mmol) in DMF (6 mL) was added t-BuOK (544 mg, 4.85 mmol) at 0 °C. After stirring at 0 °C for 2 h, 2-bromopropane (746 mg, 6.07 mmol) was added to the mixture drop-wise at 0 °C. The resulting mixture was stirred at 0 ⁇ 20 °C for 2 h. The reaction mixture was quenched by AcOH. The mixture was taken up with EtOAc (40 mL x 2) , washed with brine, dried over sodium sulfate and concentrated under reduced pressure.
  • EtOAc 40 mL x 2
  • Step 5 To a solution of methyl 2- (3- (benzyloxy) isoxazol-5-yl) -3-methylbutanoate (400 mg, 1.38 mmol) in DCM (4 mL) was added BBr 3 (692 mg, 2.76 mmol) at -78 °C. After stirring at 0 °C for 2 h, the mixture was poured into H 2 O (20 mL) . The resulting mixture was extracted with DCM (20 mL x 2) . The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column on silica gel to afford Intermediate 2 (200 mg) as a white solid. LCMS (ESI) m/z: 200.1 [M+1] + .
  • Step 1 A mixture of 2-amino-4-bromo-3-fluorobenzoic acid (2.0 g, 8.5 mmol) and urea (5.1 g, 85.5 mmol) was stirred at 200 °C for 5 h. The reaction mixture was cooled and poured into water (100 mL) . Then the resulting mixture was filtered to afford 7-bromo-8-fluoroquinazoline-2, 4-diol (2.1 g, 94.8%yield) as a yellow solid.
  • LCMS (ESI) m/z: 259.0 [M+1] + .
  • Step 2 To a solution of 7-bromo-8-fluoroquinazoline-2, 4-diol (0.5 g, 1.9 mmol) in toluene (20 mL) was added DIPEA (0.75 g, 5.8 mmol) and POCl 3 (0.9 g, 5.8 mmol) , and the reaction mixture was stirred at 110 °C for 1 h. The mixture was cooled and concentrated under reduced pressure to give 7-bromo-2, 4-dichloro-8-fluoroquinazoline (0.6 g, crude) as black oil, and used in next step directly.
  • DIPEA 0.75 g, 5.8 mmol
  • POCl 3 0.9 g, 5.8 mmol
  • Step 3 To a solution of 7-bromo-2, 4-dichloro-8-fluoroquinazoline (0.6 g, 2.0 mmol) in DCM (5 mL) was added DIPEA (0.8 g, 6.1 mmol) and tert-butyl (1R, 5S) -3, 8-diazabicyclo [3.2.1] octane-8-carboxylate (0.5 g, 2.4 mmol) at -60 °C. The reaction mixture was stirred at -60 °C for 1 h. The mixture was poured into water (10 mL) and extracted with DCM (10 mL x 3) . The combined organic phase was dried over Na 2 SO 4 and concentrated under reduced pressure.
  • Step 4 To a solution of tert-butyl (1R, 5S) -3- (7-bromo-2-chloro-8-fluoroquinazolin-4-yl) -3, 8-diazabicyclo [3.2.1] octane-8-carboxylate (4.5 g, 9.6 mmol) and 2, 2-dimethoxy ethan-1-ol (2.1 g, 19.2 mmol) in MeCN (45 mL) was added DABCO (108 mg, 0.96 mmol) and Cs 2 CO 3 (4.0 g, 12.4 mol) . The reaction mixture was stirred at 50 °C for 5 h. The reaction mixture filtered, and the filtrate was concentrated under reduced pressure.
  • Step 5 To a solution of tert-butyl (1R, 5S) -3- (7-bromo-2- (2, 2-dimethoxyethoxy) -8-fluoroquinazolin-4-yl) -3, 8-diazabicyclo [3.2.1] octane-8-carboxylate (1.1 g, 2.0 mmol) , 2- (8-ethyl-7-fluoro-3- (methoxymethoxy) naphthalen-1-yl) -4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolane (1.4 g, 4.1 mmol) and Na 2 CO 3 (754 mg, 7.1 mmol) in 1, 4-dioxane/H 2 O (30 mL/6 mL) was added Pd (PPh 3 ) 4 (234 mg, 0.2 mmol) under N 2 .
  • the reaction mixture was stirred at 110 °C for 16 h.
  • the reaction mixture was poured into water (30 mL) .
  • the aqueous phase was extracted with EtOAc (30 mL x 2) .
  • the combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 .
  • the organic layer was concentrated under reduced pressure.
  • Step 6 To a solution of tert-butyl (1R, 5S) -3- (2- (2, 2-dimethoxyethoxy) -7- (8-ethyl-7-fluoro-3- (methoxymethoxy) naphtha len-1-yl) -8-fluoroquinazolin-4-yl) -3, 8-diazabicyclo [3.2.1] octane-8-carboxylate (100 mg, 0.14 mmol) in 1, 4-dioxane (2 mL) was added aq. HCl (12 M, 0.25 mL) , the mixture was stirred at 15 °C for 2 h. Then the mixture was concentrated to afford a yellow oil.
  • Step 1 To a solution of (S) -1- (4-bromophenyl) ethan-1-amine (25 g, 0.12 mol) in DCM (250 mL) was added Boc 2 O (32.5 g, 0.15 mol) and TEA (25 g, 0.24 mol) . The reaction mixture was stirred at 25 °C for 3 h. The reaction mixture was concentrated under reduced pressure. The residue was triturated with PE (500 mL) to offer tert-butyl (S) - (1- (4-bromophenyl) ethyl) carbamate (35 g) as a white solid.
  • Step 2 To a solution of tert-butyl (S) - (1- (4-bromophenyl) ethyl) carbamate (5.0 g, 16.7 mmol) , (1-methyl-1H-pyrazol-5-yl) boronic acid (2.5 g, 20.1 mmol) and Na 2 CO 3 (2.6 g, 25.1 mmol) in 1, 4-dioxane/H 2 O (60 mL/5 mL) was added Pd (dppf) Cl 2 (1.2 g, 1.7 mmol) under N 2 . The reaction mixture was stirred at 80 °C for 16 h. The reaction mixture was poured into water (100 mL) .
  • Step 3 To a solution of tert-butyl (S) - (1- (4- (1-methyl-1H-pyrazol-5-yl) phenyl) ethyl) carbamate (3.1 g, 10.3 mmol) in DCM (20 mL) was added HCl/1, 4-dioxane (4 M, 10 mL) . The reaction mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure to afford (S) -1- (4- (1-methyl-1H-pyrazol-5-yl) phenyl) ethan-1-amine (2 g) as a yellow solid. LCMS (ESI) m/z: 202.2 [M+1] + .
  • Step 4 A solution of (2S, 4R) -1- (tert-butoxycarbonyl) -4-hydroxypyrrolidine-2-carboxylic acid (2.5 g, 10.9 mmol) , DIEA (5.1 g, 39.8 mmol) and HATU (4.5 g, 11.9 mmol) in DMF (15 mL) . The mixture was stirred at 25 °C for 15 min. Then (S) -1- (4- (1-methyl-1H-pyrazol-5-yl) phenyl) ethan-1-amine (2.0 g, 9.9 mmol) in DMF (5 mL) was added to the mixture. The reaction mixture was stirred at 25 °C for 16 h.
  • Step 5 To a solution of tert-butyl (2S, 4R) -4-hydroxy-2- ( ( (S) -1- (4- (1-methyl-1H-pyrazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidine-1-carboxylate (4.5 g, 10.8 mmol) in DCM (20 mL) was added aq. HCl (4 M, 40 mL) . The reaction mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure. The residue was triturated with EA (50 mL) to afford Intermediate 4 (4.0 g) as a brown solid. LCMS (ESI) m/z: 315.2 [M+1] + .
  • Step 1 To a solution of compound 1.1 (10 g, 1.0 eq) and TsCl (15.1 g, 1.2 eq) in DCM (50 mL) was added TEA (13.9 g, 2.1 eq) . The reaction mixture was stirred at 25 °C for 15 h. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was poured into water (200 mL) . The aqueous phase was extracted with EtOAc (200 mL x 2) . The combined organic phase was washed with brine (200 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by chromatography column on silica gel to afford compound 1.2 (18.5 g) as a yellow solid.
  • Step 2 To a solution of compound 1.2 (6.3 g, 1.05 eq) in DMF (30 mL) was added NaH (780 mg, 1.1 eq) under N 2 . The reaction mixture was stirred at 0 °C for 0.5 h. Then tert-butyl 4- (2- (tosyloxy) ethoxy) piperidine-1-carboxylate (9.0 g, 1.0 eq) was added into the mixture. The reaction mixture was stirred at 25 °C for 1 h. The mixture was poured into water (100 mL) . The aqueous phase was extracted with EtOAc (100 mL x 2) .
  • Step 3 To a solution of compound 1.3 (7.5 g, 1.0 eq) in MeOH (70 mL) was added Pd/C (750 mg, 10 w%) . The mixture was stirred at 30 °C for 12 h. The mixture was filtrated, and the filtrate was concentrated to afford the compound 1.4 (6.9 g) as a yellow solid.
  • Step 4 To a solution of compound 1.4 (886.2 mg, 1.2 eq) in THF (30 mL) was added Intermediate 2 (600 mg, 1.0 eq) and PPh 3 (948.2 mg, 1.2 eq) . The mixture was stirred at 0 °C for 10 min. Then DIAD (730.5 mg, 1.2 eq) was added into the mixture. The mixture was stirred at 30 °C for 16 h. The reaction mixture was poured into water (20 mL) . The aqueous phase was extracted with EtOAc (20 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by chromatography column on silica gel to afford compound 1.5 (1.1 g) as a yellow solid. LCMS (ESI) m/z: 327.2 [M-100+1] + .
  • Step 5 To a solution of compound 1.5 (800 mg, 1.0 eq) in THF/H 2 O/MeOH (4 mL/2 mL/2 mL) was added LiOH (71.7 mg, 1.5 eq) . The reaction mixture was stirred at 25 °C for 2 h. The pH of the reaction mixture was adjusted to 5 with 1 M aq. HCl. The aqueous phase was extracted with EtOAc (10 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure to afford compound 1.6 (850 mg) as a white solid. LCMS (ESI) m/z: 313.2, [M-100+1] + .
  • Step 6 To a solution of compound 1.6 (850 mg, 1.0 eq) , intermediate 1 (719.4 mg, 1.1 eq) and HATU (1.2 g, 1.5 eq) in DMF (10 mL) was added DIEA (799.9 mg, 3.0 eq) . The reaction mixture was stirred at 25 °C for 2 h. The reaction mixture was poured into water (30 mL) . The aqueous phase was extracted with EtOAc (40 mL x 3) . The combined organic phase was washed with brine (50 mL x 2) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by chromatography column on silica gel to afford compound 1.7 (1.0 g) as a white solid. LCMS (ESI) m/z: 726.3 [M+1] + .
  • Step 7 To a solution of compound 1.7 in DCM (5 mL) was added HCl/dioxane (4 M, 2 mL) . The mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure to afford compound 1.8 (503 mg, crude) as a yellow solid.
  • Step 8 To a solution of compound 1.8 (110 mg, 1.0 eq) in MeOH (4 mL) was added NaOAc (27.2 mg, 2.0 eq) . The mixture was stirred at 25 °C for 10 min. Then intermediate 3 (100 mg, 1.0 eq) and NaBH 3 CN (40.8 mg, 4.0 eq) was added. The mixture was stirred at 25 °C for 16 h. The reaction mixture was poured into water (20 mL) . The aqueous phase was extracted with EtOAc (20 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by chromatography column on silica gel to afford compound 1.9 (102 mg) as a yellow solid. LCMS (ESI) m/z: 607.8 [1/2M+1] + .
  • Step 9 To a solution of compound 1.9 (102 mg, 1.0 eq) in DCM (5 mL) was added HCl/1, 4-dioxane (4 M, 2 mL) . The reaction mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (FA) to afford Example 1 (42.2 mg, 1.37 FA salt, 99.4%HPLC purity) as a white solid. LCMS (ESI) m/z: 1116.4 [M+1] + .
  • Step 1 To a solution of Intermediate 2 (65 mg, 0.32 mmol) and K 2 CO 3 (22 mg, 0.16 mmol) in MeCN (5 mL) was added 1, 1, 2, 2, 3, 3, 4, 4, 4-nonafluorobutane-1-sulfonyl fluoride (197 mg, 0.65 mmol) at 0 °C under N 2 . The reaction mixture was stirred at 20 °C for 16 h. The mixture was diluted with aq. NH 4 Cl (20 mL) and extracted with EtOAc (30 mL) . The organic phase was washed with brine (20 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure to afford compound 2.1 (100 mg) as a yellow solid.
  • Step 2 A solution of compound 2.1 (100 mg, 0.21 mmol) , tert-butyl 2, 7-diazaspiro [3.5] nonane-7-carboxylate (47 mg, 0.21 mmol) and DIPEA (80 mg, 0.62 mmol) in DMAC (2 mL) was stirred at 140 °C for 4 h. Then the mixture was diluted with water (20 mL) and extracted with EtOAc (20 mL) . The organic phase was washed with brine (20 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by chromatography column on silica gel to afford compound 2.2 (20 mg) as a yellow oil. LCMS (ESI) m/z: 408.3 [M+1] + .
  • Step 3 To a solution of compound 2.2 (260 mg, 0.64 mmol) in THF (2.5 mL) , MeOH (2.5 mL) and water (2.5 mL) was added LiOH. H 2 O (133 mg, 3.18 mmol) . The reaction mixture was stirred at 25 °C for 1.5 h. The pH of the mixture was adjusted to ⁇ 3 with aq. citric acid (10 w%) . Then the mixture was extracted with DCM (20 mL) . The organic phase was washed with brine (20 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure to afford compound 2.3 (260 mg) as yellow solid. LCMS (ESI) m/z: 394.3 [M+1] + .
  • Step 4 To a solution of compound 2.3 (260 mg, 0.66 mmol) , Intermediate 1 (242 mg, 0.66 mmol) , HATU (301 mg, 0.79 mmol) in DMF (5 mL) was added DIPEA (426 mg, 3.30 mmol) . The mixture was stirred at 25 °C for 1 h. The reaction mixture was poured into water (20 mL) . The aqueous phase was extracted with EtOAc (20 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by chromatography column on silica gel to afford compound 2.4 (290 mg) as a yellow solid. LCMS (ESI) m/z: 707.3 [M+1] + .
  • Step 5 To a solution of compound 2.4 (290 mg, 0.41 mmol) in DCM (6 mL) was added TFA (2 mL) , the mixture was stirred at 15 °C for 2 h. The mixture was diluted with aq. NaHCO 3 (20 mL) and extracted with DCM (20 mL) . The organic phase was concentrated under reduced pressure to afford compound 2.5 (190 mg) as a yellow solid.
  • Step 6 To a solution of compound 2.5 (80 mg, 0.13 mmol) , intermediate 3 (80 mg, 0.13 mmol) in MeOH (4 mL) was added NaBH 3 CN (25 mg, 0.39 mmol) . The mixture was stirred at 25 °C for 16 h. The reaction mixture was poured into water (20 mL) . The aqueous phase was extracted with EtOAc (20 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by chromatography column on silica gel to afford compound 2.6 (60 mg) as a yellow solid. LCMS (ESI) m/z: 399.4 [1/2M+1] + .
  • Step 7 To a solution of compound 2.6 (60 mg, 0.05 mmol) in DCM (3 mL) was added TFA (1.0 mL) . The reaction mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (FA) to afford Example 2 (36.3 mg, 0.89 FA salt, 99.9%HPLC purity) as a white solid. LCMS (ESI) m/z: 548.3 [1/2M+1] + .
  • Step 1 To a solution of intermediate 2 (500 mg, 2.5 mmol) , compound 3.1 (648 mg, 3.0 mmol) and PPh 3 (790 mg, 3.0 mmol) in anhydrous THF (10 mL) was added DIAD (610 mg, 3.0 mmol) at 0 °C under N 2 , the reaction mixture was stirred at 25 °Cfor 3 h. The reaction mixture was poured into water (30 mL) . The aqueous phase was extracted with EtOAc (30 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure.
  • DIAD 610 mg, 3.0 mmol
  • Step 2 To a solution of compound 3.2 (890 mg, 2.2 mmol) in THF/H 2 O/MeOH (4 mL/4 mL/4 mL) was added LiOH (141 mg, 3.4 mmol) . The mixture was stirred at 25 °C for 2 h. The pH of the reaction mixture was adjusted to ⁇ 5 with aq. HCl (1 M) . The aqueous phase was extracted with EtOAc (10 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure to afford compound 3.3 (820 mg) as a white solid.
  • Step 3 To a solution of compound 3.3 (820 mg, 2.1 mmol) , intermediate 1 (680 mg, 2.1 mmol) and HATU (1.2 g, 3.2 mmol) in DMF (15 mL) was added DIEA (828 mg, 6.4 mmol) . The reaction mixture was stirred at 25 °C for 2 h. The reaction mixture was poured into water (30 mL) . The aqueous phase was extracted with EtOAc (40 mL x 3) . The combined organic phase was washed with brine (50 mL x 2) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by chromatography column on silica gel to afford the compound 3.4 (1.2 g) as yellow solid. LCMS (ESI) m/z: 696.3 [M+1] + .
  • Step 4 To a solution of compound 3.4 (600 mg, 0.86 mmol) in DCM (10 mL) was added HCl/1, 4-dioxane (4 M, 5 mL) . The mixture was stirred at 25 °C for 1 h. The mixture was concentrated under reduced pressure to afford compound 3.5 (560 mg, crude) as a yellow solid.
  • Step 5 To a solution of compound 3.5 (100 mg, 0.16 mmol) in MeOH (4 mL) was added NaOAc (27 mg, 0.33 mmol) . The mixture was stirred at 25 °C for 10 min. Then intermediate 3 (100 mg, 0.16 mmol) and NaBH 3 CN (25 mg, 0.40 mmol) was added into the mixture. The mixture was stirred at 25 °C for 16 h. The reaction mixture was poured into water (20 mL) . The aqueous phase was extracted with EtOAc (20 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by chromatography column on silica gel to afford compound 3.6 (62 mg) as a yellow solid. LCMS (ESI) m/z: 592.9 [1/2M+1] + .
  • Step 6 To a solution of compound 3.6 (62 mg, 0.052 mmol) in DCM (1 mL) was added HCl/1, 4-dioxane (4 M, 1 mL) . The reaction mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (FA) to afford Example 3 (43.4 mg, 0.89 FA salt) as a white solid.
  • Step 1 To a solution of compound 4.1 (600 mg, 1.0 eq) , intermediate 2 (843 mg, 1.2 eq) and PPh 3 (572.4 mg, 1.2 eq) in anhydrous THF (10 mL) was added DIAD (731.4 mg, 1.2 eq) at 0 °C under N 2 . The reaction mixture was stirred at 80 °C for 3 h. The reaction mixture was poured into water (30 mL) . The aqueous phase was extracted with EtOAc (30 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by chromatography column on silica gel to afford compound 4.2 (253 mg) as a white solid. LCMS (ESI) m/z: 316.2 [M+1] + .
  • Step 2 To a solution of compound 4.2 (253 mg, 1.0 eq) in THF/H 2 O/MeOH (4 mL/2 mL/2 mL) was added LiOH (25 mg, 1.5 eq) . The reaction mixture was stirred at 25 °C for 2 h. The pH of the reaction mixture was adjusted to ⁇ 5 with aq. HCl (1 M) . The aqueous phase was extracted with EtOAc (10 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure to afford compound 4.3 (245 mg) as a white solid. LCMS (ESI) m/z: 301.2 [M-100+1] + .
  • Step 3 To a solution of compound 4.3 (245 mg, 1.0 eq) , intermediate 4 (252.5 mg, 1.5 eq) and HATU (320.2 mg, 1.5 eq) in DMF (10 mL) was added DIPEA (219.5 mg, 3.0 eq) . The reaction mixture was stirred at 25 °C for 2 h. The reaction mixture was poured into water (30 mL) . The aqueous phase was extracted with EtOAc (40 mL x 3) . The combined organic phase was washed with brine (50 mL x 2) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by chromatography column on silica gel to afford compound 4.4 (490 mg) as yellow solid. LCMS (ESI) m/z: 697.3 [M+1] + .
  • Step 4 To a solution of compound 4.4 (490 mg, 1.0 eq) in DCM (5 mL) was added HCl/1, 4-dioxane (4 M, 2 mL) . The mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure to afford compound 4.5 (503 mg, crude) as a yellow solid.
  • Step 5 To a solution of compound 4.5 (100 mg, 1.0 eq) in MeOH (4 mL) was added NaOAc (27.2 mg, 2.0 eq) . The mixture was stirred at 25 °C for 10 min. Then intermediate 3 (108.6 mg, 1.1 eq) and NaBH 3 CN (41.6 mg, 4.0 eq) was added into the mixture. The mixture was stirred at 25 °C for 16 h. The reaction mixture was poured into water (20 mL) . The aqueous phase was extracted with EtOAc (20 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by chromatography column on silica gel to afford compound 4.6 (103 mg) as a yellow solid. LCMS (ESI) m/z: 593.5 [1/2M+1] + .
  • Step 6 To a solution of compound 4.6 (103 mg, 1.0 eq) in DCM (5 mL) was added HCl/1, 4-dioxane (4 M, 2 mL) . The reaction mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (FA) to Example 4 (45.6 mg, 0.81 FA salt, 98.3%HPLC purity) as a white solid. LCMS (ESI) m/z: 543.4 [1/2M+1] + .
  • Step 1 To a solution of intermediate 2 (500 mg, 1.0 eq) , compound 5.1 (688.5 mg, 1.2 eq) and PPh 3 (895.1 mg, 1.2 eq) in anhydrous THF (10 mL) was added DIAD (609.5 mg, 1.2 eq) at 0 °C under N 2 . The reaction mixture was stirred at 75 °C for 3 h. The reaction mixture was poured into water (30 mL) . The aqueous phase was extracted with EtOAc (30 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by chromatography column on silica gel to afford compound 5.2 (1.0 g) as a white solid. LCMS (ESI) m/z: 311.2 [M+1] + .
  • Step 2 To a solution of compound 5.2 (500 mg, 1.0 eq) in THF/H 2 O/MeOH (2 mL/1 mL/1 mL) was added LiOH (143.9 mg, 1.5 eq) . The reaction mixture was stirred at 25 °C for 2 h. The pH of the reaction mixture was adjusted to 5 with 1 M aq. HCl. The aqueous phase was extracted with EtOAc (10 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure to afford compound 5.3 (500 mg) as a white solid. LCMS (ESI) m/z: 297.2 [M-100+1] + .
  • Step 3 To a solution of compound 5.3 (500 mg, 1.0 eq) , intermediate 1 (480.3 mg, 1.2 eq) and HATU (720.2 mg, 1.5 eq) in DMF (15 mL) was added DIPEA (489.5 mg, 3.0 eq) . The reaction mixture was stirred at 25 °C for 2 h. The reaction mixture was poured into water (30 mL) . The aqueous phase was extracted with EtOAc (40 mL x 3) . The combined organic phase was washed with brine (50 mL x 2) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by chromatography column on silica gel to afford compound 5.4 (740 mg) as yellow solid. LCMS (ESI) m/z: 710.3 [M+1] + .
  • Step 4 To a solution of compound 5.4 (300 mg, 1.0 eq) in DCM (5 mL) was added HCl/1, 4-dioxane (4 M, 2 mL) . The mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure to afford compound 5.5 (300 mg, crude) as a yellow solid. LCMS (ESI) m/z: 610.3 [M+1] + .
  • Step 5 To a solution of compound 5.5 (106 mg, 1.0 eq) in MeOH (4 mL) was added NaOAc (27.2 mg, 2.0 eq) . The mixture was stirred at 25 °C for 10 min. Then intermediate 3 (100 mg, 1.0 eq) and NaBH 3 CN (42 mg, 4.0 eq) were added into the mixture. The mixture was stirred at 25 °C for 16 h. The reaction mixture was poured into water (20 mL) . The aqueous phase was extracted with EtOAc (20 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by chromatography column on silica gel to afford compound 5.6 (110 mg) as a yellow solid. LCMS (ESI) m/z: 499.2 [1/2M+1] + .
  • Step 6 To a solution of compound 5.6 (110 mg, 1.0 eq) in DCM (5 mL) was added HCl/1, 4-dioxane (4 M, 2 mL) . The reaction mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (FA) to Example 5 (53.9 mg, 1.38 FA salt, 99.3%HPLC purity) as a white solid. LCMS (ESI) m/z: 1099.4 [M+1] + .
  • Step 1 A solution of tert-butyl (1R, 5S) -3- (7-bromo-2-chloro-8-fluoroquinazolin-4-yl) -3, 8-diazabicyclo [3.2.1] octane-8-ca rboxylate (3.0 g, 6.4 mmol) and KF (2.9 g, 50.8 mmol) in DMSO (30 mL) was stirred at 120 °C for 16 h. The reaction mixture was poured into water. The aqueous phase was extracted with EtOAc. The combined organic phase was washed with brine and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel to afford compound 7.1 (2.4 g) as a yellow solid. LCMS (ESI) m/z: 457.1 [M+1] + .
  • Step 2 To a solution of (3- ( ( (tert-butyldimethylsilyl) oxy) methyl) tetrahydro-1H-pyrrolizin-7a (5H) -yl) methano l (1.4 g, 4.9 mmol) in THF (10 mL) was added NaH (264 mg, 6.6 mmol, 60 w%) at 0 °C under N 2 . The mixture was stirred at 0 °C for 30 min. Then compound 7.1 (1.5 g, 3.3 mmol) in THF (5 mL) was added. The reaction mixture was stirred at 30 °C for 5 h. The reaction mixture was quenched by aq. NH 4 Cl.
  • Step 3 To a solution of compound 7.2 (1.0 g, 1.4 mmol) in THF (10 mL) was added TBAF (8.3 mL, 1 mol/L in THF) . The reaction mixture was stirred at 30 °C for 2 h. The reaction mixture was poured into water. The aqueous phase was extracted with EtOAc. The combined organic phase was washed with brine and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel to afford compound 7.3 (750 mg) as yellow oil. LCMS (ESI) m/z: 609.3 [M+1] + .
  • Step 4 To a solution of compound 7.3 (750 mg, 1.2 mmol) , intermediate 2 (270 mg, 1.4 mmol) and PPh 3 (486 mg, 1.8 mmol) in anhydrous THF (10 mL) was added DIAD (375 mg, 1.8 mmol) under N 2 . The reaction mixture was stirred at 40 °C for 3 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel to afford compound 7.4 (800 mg) as yellow oil. LCMS (ESI) m/z: 789.3 [M+1] + .
  • Step 5 A solution of compound 7.4 (400 mg, 0.51 mmol) , 2- (8-ethyl-7-fluoro-3- (methoxymethoxy) naphthalen-1-yl) -4, 4, 5, 5-tetramethyl-1, 3, 2-di oxaborolane (366 mg, 1.0 mmol) , Pd (PPh 3 ) 4 (50 mg, 0.05 mmol) and Na 2 CO 3 (188 mg, 1.8 mmol) in 1, 4-dioxane/H 2 O (10 /2 mL) was stirred at 120 °C for 16 h under N 2 . The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-TLC and prep-HPLC (TFA) to afford compound 7.5 (49 mg) as a yellow solid. LCMS (ESI) m/z: 471.4 [1/2M+1] + .
  • Step 6 To a solution of compound 7.5 (49 mg, 0.052 mmol) in THF/H 2 O (2/2 mL) was added LiOH (6 mg, 0.26 mmol) . The reaction mixture was stirred at 25 °C for 2 h. The pH of the reaction mixture was adjusted to ⁇ 5 with aq. HCl (1 M) . The aqueous phase was extracted with EtOAc. The combined organic phase was washed with brine and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure to afford compound 7.6 (49 mg, crude) as yellow oil. LCMS (ESI) m/z: 464.3 [1/2M+1] + .
  • Step 7 To a solution of compound 7.6 (49 mg, 0.052 mmol) and (2S, 4R) -4-hydroxy-N- ( (S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carb oxamide (18 mg, 0.052 mmol) in DMF (2 mL) was added HATU (30 mg, 0.079 mmol) and DIPEA (20 mg, 0.15 mmol) . The reaction mixture was stirred at 25 °C for 2 h. The residue was poured into water. The aqueous phase was extracted with EtOAc. The combined organic phase was washed with brine and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by prep-HPLC to give compound 7.7 (18 mg) as yellow oil.
  • Step 8 To a solution of compound 7.7 (18 mg, 0.014 mmol) in DCM (2 mL) was added HCl/1, 4-dioxane (4 M, 2 mL) . The reaction mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (FA) to afford Example 7 (2.8 mg, 90%HPLC purity, 0.7 FA salt) as a white solid. Chemical Formula: C 60 H 67 F 2 N 9 O 7 S. Molecular Weight: 1096.31. LCMS (ESI) m/z: 1096.6 [M+1] + .
  • Step 1 To a solution of tert-butyl (1R, 5S) -3- (7-bromo-2- (2, 2-dimethoxyethoxy) -8-fluoroquinazolin-4-yl) -3, 8-diazabicyclo [3.2.1] octane-8-carboxylate (100 mg, 0.18 mmol) , ( (2-fluoro-6- (methoxymethoxy) -8- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) napht halen-1-yl) ethynyl) triisopropylsilane (199 mg, 0.39 mmol) and Na 2 CO 3 (69 mg, 0.55 mmol) in 1, 4-dioxane/H 2 O (10 mL/2 mL) was added Pd (PPh 3 ) 4 (21 mg, 0.018 mmol) under N 2 .
  • reaction mixture was stirred at 110 °C for 16 h.
  • the reaction mixture was poured into water (30 mL) .
  • the aqueous phase was extracted with EtOAc (30 mL x 2) .
  • the combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 .
  • the organic layer was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel to afford compound 31.1 (60 mg) as a yellow gum.
  • Step 2 To a solution of compound 31.1 (60 mg, 0.087 mmol) in DMF (10 mL) was added CsF (20 mg, 0.13 mmol) . The reaction mixture was stirred at r.t. for 2 h. The reaction mixture was poured into water (30 mL) . The aqueous phase was extracted with EtOAc (30 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel to afford compound 31.2 (49.4 mg) as a brown gum. LCMS (ESI) m/z: 691.8 [M+1] + .
  • Step 3 To a solution of compound 31.2 (49.4 mg, 0.087 mmol) in 1, 4-dioxane (10 mL) was added aq. HCl (12 M, 1 mL) . The mixture was stirred at 0 °C for 2 h. To the mixture was added THF (30 mL) and H 2 O (30 mL) . Then NaHCO 3 was added to adjusted pH to 8 ⁇ 9. Then Boc 2 O (19 mg, 0.087 mmol) was added. The mixture was stirred at r.t. for 1 h. The reaction was diluted with water (10 mL) and extracted with EtOAc (20 mL) .
  • Step 4 To a solution of compound 31.3 (38.8 mg, 1.5 eq) in MeOH (30 mL) was added KOAc (8.5 mg, 2.0 eq) . The mixture was stirred at 25 °C for 10 min. Then the mixture was added compound 4.4-R (27.4 mg, 1.0 eq) , NaBH 3 CN (11 mg, 4.0 eq) . The mixture was stirred at 25 °C for 16 h. The reaction mixture was poured into water (20 mL) . The aqueous phase was extracted with EtOAc (20 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel to afford compound 31.4 (19.2 mg) as a yellow solid. LCMS (ESI) m/z: 591.9 [1/2M+1] + .
  • Step 5 To a solution of compound 31.4 (19.2 mg, 1.0 eq) in DCM (10 mL) was added HCl/1, 4-dioxane (4 M, 5 mL) . The reaction mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (FA) to afford Example 31 (9.9 mg, 97.65%HPLC purity) as a white solid.
  • Step 1 To a solution of 7-bromo-2, 4-dichloro-6, 8-difluoroquinazoline (12.2 g, 9.6 mmol) and tert-butyl (1R, 5S) -3, 8-diazabicyclo [3.2.1] octane-8-carboxylate (9.9 g, 14.4 mmol) in DCM (120 mL) was added DIPEA (15 g, 116.6 mmol) at -70 °C. The reaction mixture was stirred at -70 °C for 2 h. The mixture was poured into ice-water (300 mL) . The aqueous phase was extracted with EtOAc (100 mL x 2) .
  • Step 2 To a solution of compound 67.1 (4.7 g, 9.6 mmol) and 2, 2-dimethoxyethan-1-ol (1.5 g, 14.4 mmol) in MeCN (70 mL) was added DABCO (108 mg, 0.96 mmol) and Cs 2 CO 3 (9.4 g, 28.8 mmol) . The reaction mixture was stirred at 50 °C for 5 h. The reaction mixture filtered, and the filtrate was concentrated under reduced pressure. The residue was poured into water (50 mL) . The aqueous phase was extracted with EtOAc (40 mL x 2) . The combined organic phase was washed with brine (50 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel to afford compound 67.2 (5 g) as a yellow solid. LCMS (ESI) m/z: 561.5 [M+1] + .
  • Step 3 To a solution of compound 67.2 (2.8 g, 5.0 mmol) , ( (2-fluoro-6- (methoxymethoxy) -8- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) napht halen-1-yl) ethynyl) triisopropylsilane (5.4g, 10.5 mmol) and Na 2 CO 3 (1.86 g, 15.0 mmol) in 1, 4-dioxane/H 2 O (100 mL/20 mL) was added G3-Pd (365 mg, 0.5 mmol) under N 2 . The mixture was stirred at 110 °C for 16 h.
  • Step 4 To a solution of compound 67.3 (3.63 g, 4.2 mmol) in DMF (50 mL) was added CsF (638 mg, 4.2 mmol) . The reaction mixture was stirred at r.t. for 2 h. The reaction mixture was poured into water (200 mL) . The aqueous phase was extracted with EtOAc (50 mL x 2) . The combined organic phase was washed with brine (100 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel to afford compound 67.4 (3.04 g) as a brown gum. LCMS (ESI) m/z: 709.8 [M+1] + .
  • Step 5 To a solution of compound 67.4 (3.04 g, 4.29 mmol) in 1, 4-dioxane (60 mL) was added aq. HCl (12 M, 7.5 mL) . The mixture was stirred at 0 °C for 2 h. Then to the mixture was added THF (30 mL) and H 2 O (30 mL) . Then NaHCO 3 was added to adjusted the pH to 8 ⁇ 9. Then Boc 2 O (936 mg, 4.29 mmol) was added. The mixture was stirred at r.t. for 1 h. The reaction was diluted with water (10 mL) and extracted with EtOAc (20 mL) .
  • Step 6 To a solution of compound 4.4-R (1.08 g, 1.1 eq) in MeOH (30 mL) was added KOAc (323 mg, 2.0 eq) , the mixture was stirred at 25 °C for 10 min. Then compound 67.5 (1.02 g, 1.0 eq) and NaBH 3 CN (415 mg, 4.0 eq) were added. The mixture was stirred at 25 °C for 16 h. The reaction mixture was poured into water (20 mL) . The aqueous phase was extracted with EtOAc (20 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel to afford compound 67.6 (880 mg) as a yellow solid. LCMS (ESI) m/z: 600.5 [1/2M+1] + .
  • Step 7 To a solution of compound 67.6 (880 mg, 1.0 eq) in DCM (10 mL) was added HCl/1, 4-dioxane (4 M, 5 mL) . The reaction mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (FA) to afford Example 67 (278 mg, 94%HPLC purity) as a white solid. LCMS (ESI) m/z: 550.8 [1/2M+1] + .
  • Step 1 To a solution of 6-chloro-5-fluoro-2, 7-naphthyridine-1, 3 (2H, 4H) -dione (200 mg, 0.93 mmol) in POCl 3 was added DIPEA (360 mg, 2.8 mmol) . The reaction mixture was stirred at 100 °C overnight. The reaction mixture filtered, and the filtrate was concentrated under reduced pressure. The residue was diluted with DCM, then sat. aq. NaHCO 3 (50 mL) was added drop-wise. The aqueous phase was extracted with DCM (40 mL x 2) . The combined organic phase was washed with brine (50 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel to afford compound 115.1 (108 mg, crude) as a yellow solid. LCMS (ESI) m/z: 252.1 [M+1] + .
  • Step 2 To a solution of compound 115.1 (220 mg, 0.87 mmol) in DCM was added Et 3 N (275 mg, 2.7 mmol) . The mixture was cooled to -60 °C, then tert-butyl (1R, 5S) -3, 8-diazabicyclo [3.2.1] octane-8-carboxylate (186 mg, 0.87 mmol) was added. The resulting mixture was stirred for 30 min. The mixture was added into sat. aq. NH 4 Cl drop-wise. The aqueous phase was extracted with DCM (40 mL x 2) . The combined organic phase was washed with brine (50 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel to afford compound 115.2 (350 mg) as a yellow solid. LCMS (ESI) m/z: 428.4 [M+1] + .
  • Step 3 To a solution of compound 115.2 (50 mg, 0.12 mmol) and 2, 2-dimethoxyethan-1-ol (24.8 mg, 0.23 mmol) in MeCN (1 mL) was added DABCO (1.31 mg, 0.012 mmol) and Cs 2 CO 3 (49.5 mg, 0.15 mmol) . The reaction mixture was stirred at 50 °C for 5 h. The reaction mixture filtered, and the filtrate was concentrated under reduced pressure. The residue was poured into water (50 mL) . The aqueous phase was extracted with EtOAc (40 mL x 2) . The combined organic phase was washed with brine (50 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel to afford compound 115.3 (61 mg) as a yellow solid. LCMS (ESI) m/z: 498.6 [M+1] + .
  • Step 4 To a solution of compound 115.3 (427 mg, 0.85 mmol) , ( (2-fluoro-6- (methoxymethoxy) -8- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) napht halen-1-yl) ethynyl) triisopropylsilane (924.4 mg, 1.8 mmol) and Cs 2 CO 3 (980 mg, 3.0 mmol) in 1, 4-dioxane/H 2 O (4 mL/0.8 mL) was added Pd-G3 (62.6 mg, 0.085 mmol) under N 2 . The reaction mixture was stirred at 110 °C for 5 h.
  • Step 5 To a solution of compound 115.4 (551 mg, 0.65 mmol) in DMF (8 mL) was added CsF (99.0 mg, 0.65 mmol) . The reaction mixture was stirred at r.t. for 2 h. The reaction mixture was poured into water (30 mL) . The aqueous phase was extracted with EtOAc (30 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure to afford the compound 115.5 (535 mg) as a yellow gum. LCMS (ESI) m/z: 692.9 [M+1] + .
  • Step 6 To a solution of compound 115.5 (50 mg, 0.07 mmol) in 1, 4-dioxane (2 mL) was added aq. HCl (12 M, 0.25 mL) . The mixture was stirred at 15 °C for 2 h. Then the mixture was concentrated. THF (2 mL) and H 2 O (1 mL) were added into the residue. Then NaHCO 3 (30 mg, 0.042 mmol) and Boc 2 O (33.5 mg, 0.15 mmol) were added. The mixture was stirred at 15 °C for 1.5 h. The reaction was diluted with water (10 mL) and extracted with DCM (20 mL) .
  • Step 7 To a solution of tert-butyl (1R, 5S) -3- (7- (8-ethynyl-7-fluoro-3-hydroxy naphthalen-1-yl) -8-fluoro-2- (2-oxoethoxy) pyrido [4, 3-d] pyrimidin-4-yl) -3, 8-diazabicy clo [3.2.1] octane-8-carboxylate (407 mg, 0.65 mmol) in MeOH (10 mL) was added KOAc (128.7 mg, 1.31 mmol) . The mixture was stirred at 25 °C for 30 min.
  • Step 8 To a solution of compound 115.7 (520 mg, 1.0 eq) in 1, 4-dioxane (5 mL) was added HCl/1, 4-dioxane (4 M, 7 mL) . The mixture was stirred at 25 °C for 2 h. Then the mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (FA) to afford Example 115 (104.2 mg, 92%HPLC purity and 90.7 mg, 90%HPLC purity, 2.87 FA salt) as a yellow solid. LCMS (ESI) m/z: 542.4 [1/2M+1] + .
  • Step 1 To a solution of compound 115.3 (50 mg, 0.1 mmol) , 2- (8-ethyl-7-fluoro-3- (methoxymethoxy) naphthalen-1-yl) -4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolane (76 mg, 0.21 mmol) and Cs 2 CO 3 (125 mg, 0.35 mmol) in 1, 4-dioxane/H 2 O (7 mL/2.5 mL) was added Pd (PPh 3 ) 4 (11.6 mg, 0.01 mmol) under N 2 . The reaction mixture was stirred at 140 °C for 2 h. The mixture was poured into water (30 mL) .
  • Step 2 To a solution of compound 122.1 (59 mg, 0.08 mmol) in 1, 4-dioxane (3 mL) was added aq. HCl (12 M, 0.7 mL) . The mixture was stirred at 25 °C for 30 minutes. Then the mixture was concentrated to afford a yellow oil. To a solution of the yellow oil in THF (2 mL) and H 2 O (1 mL) was added sat. aq. NaHCO 3 (6 mL) and Boc 2 O (34.8 mg, 0.16 mmol) . The mixture was stirred at 25 °C for 1 h. The reaction was diluted with water (10 mL) and extracted with DCM (20 mL) .
  • Step 3 To a solution of compound 122.2 (41 mg, 0.66 mmol) in MeOH (2.5 mL) was added KOAc (12.9 mg, 0.13 mmol) . The mixture was stirred at 25 °C for 30 min. Then compound 4.4-R (39.2 mg, 0.65 mmol) and NaBH 3 CN (16.6 mg, 0.263 mmol) were added. The mixture was stirred at 25 °C for 1 h. The reaction mixture was poured into water (20 mL) . The aqueous phase was extracted with EtOAc (20 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel to afford compound 122.3 (42.0 mg) as an oil. LCMS (ESI) m/z: 594.5 [1/2M+1] + .
  • Step 4 To a solution of compound 122.3 (42.0 mg, 1.0 eq) in 1, 4-dioxane (2.5 mL) was added HCl/1, 4-dioxane (4 M, 0.6 mL) . The reaction mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (FA) to afford Example 122-1 (30 mg, 95%HPLC purity, 3.95 FA salt) as a white solid. LCMS (ESI) m/z: 544.4 [1/2M+1] + .
  • Step 1 To a solution of compound 145.1 (1.0 g, 4.1 mmol) in THF (10 mL) was added LiHMDS (1 M, 5.3 mL, 5.3 mmol) at -78 °C. The mixture was stirred at -78 °Cfor 1 h. Then 2-iodopropane (1.1 g, 6.6 mmol) was added at -78 °C. The reaction mixture was stirred at 25 °C for 2 h. The reaction mixture was cooled to 0 °C and quenched with sat. aq. NH 4 Cl (30 mL) . The aqueous phase was extracted with EtOAc (30 mL x 2) .
  • Step 2 To a solution of LiAlH 4 (133 mg, 3.5 mmol) in THF (5 mL) was added compound 145.2 (500 mg, 1.7 mmol) at 0 °C under N 2 . The mixture was stirred at 25 °C for 4 h. The reaction mixture was cooled to 0 °C, quenched by addition of H 2 O (10 mL) , followed by aq. NaOH (10 mL, 15 w%) and H 2 O (30 mL) . After being stirred at room temperature for 10 min, the mixture was removed by filtration. The filtrate was extracted with EtOAc (30 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 .
  • Step 3 A solution of compound 145.3 (100 mg, 0.50 mmol) , intermediate 2 (150 mg, 0.60 mmol) and PPh 3 (158 mg, 0.60 mmol) in Toluene (2 mL) was stirred at 90 °C for 10 min. Then DIAD (122 mg, 0.60 mmol) was added at 90 °C under N 2 . The reaction mixture was stirred at 90 °C for 16 h. The reaction mixture was poured into water (20 mL) . The aqueous phase was extracted with EtOAc (20 mL x 2) . The combined organic phase was washed with brine (20 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure.
  • Step 4 To a solution of compound 145.4 (150 mg, 0.34 mmol) in THF/H 2 O/MeOH (2/1/1 mL) was added LiOH (13 mg, 0.51 mmol) . The reaction mixture was stirred at 25 °C for 2 h. The pH of the reaction mixture was adjusted to 5 with aq. HCl (1 M) . The aqueous phase was extracted with EtOAc (10 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure to afford compound 145.5 (145 mg) as yellow oil. Chemical Formula: C 22 H 36 N 2 O 6 . Molecular Weight: 424.54. LCMS (ESI) m/z: 325.2 [M-100+1] + .
  • Step 5 To a solution of compound 145.5 (158 mg, 0.37 mmol) , intermediate 4 (150 mg, 0.24 mmol) and HATU (202 mg, 0.37 mmol) in DMF (10 mL) was added DIEA (136 mg, 0.74 mmol) . The reaction mixture was stirred at 25 °C for 2 h. The reaction mixture was poured into water (30 mL) . The aqueous phase was extracted with EtOAc (40 mL x 3) . The combined organic phase was washed with brine (50 mL x 2) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure.
  • Step 6 To a solution of compound 145.6 (190 mg, 0.26 mmol) in DCM (2 mL) was added HCl/1, 4-dioxane (4 M, 2 mL) . The mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure to afford compound 145.7 (170 mg, crude) as a yellow solid. Chemical Formula: C 34 H 48 N 6 O 5 . Molecular Weight: 620.79. LCMS (ESI) m/z: 621.3 [M+1] + .
  • Step 7 To a solution of intermediate 3 (40 mg, 0.066 mmol) in MeOH (4 mL) was added NaOAc (10.8 mg, 0.13 mmol) . The mixture was stirred at 25 °C for 10 min. Then compound 145.7 (45 mg, 0.072 mmol) and NaBH 3 CN (16.8 mg, 0.26 mmol) were added. The mixture was stirred at 25 °C for 16 h. The reaction mixture was poured into water (20 mL) . The aqueous phase was extracted with EtOAc (20 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure.
  • Step 8 To a solution of compound 145.8 (50 mg, 0.041 mmol) in DCM (5 mL) was added HCl/1, 4-dioxane (4 M, 2 mL) . The reaction mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (FA) to afford Example 145 (20.8 mg, 99%HPLC purity, 0.9 FA salt) as a white solid. Chemical Formula: C 62 H 74 F 2 N 10 O 7 . Molecular Weight: 1109.33. LCMS (ESI) m/z: 555.3 [1/2M+1] + .
  • Step 1 To a solution of intermediate 2 (180 mg, 0.91 mmol) , compound 146.1 (250 mg, 1.1 mmol) and PPh 3 (290 mg, 1.1 mmol) in THF (10 mL) was added DIAD (223 mg, 1.1 mmol) at 0 °C under N 2 . The reaction mixture was stirred at 25 °C for 4 h. The reaction mixture was poured into water (30 mL) . The aqueous phase was extracted with EtOAc (30 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure.
  • Step 2 To a solution of compound 146.2 (150 mg, 0.37 mmol) in THF/H 2 O/MeOH (4/2/2 mL) was added LiOH (23 mg, 0.54 mmol) . The reaction mixture was stirred at 25 °C for 2 h. The pH of the reaction mixture was adjusted to 5 with aq. HCl (1 M) . The aqueous phase was extracted with EtOAc (10 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure to afford compound 146.3 (140 mg) as yellow oil. Chemical Formula: C 20 H 30 N 2 O 6 . Molecular Weight: 394.47. LCMS (ESI) m/z: 295.1 [M-100+1] + .
  • Step 3 To a solution of compound 146.3 (140 mg, 0.35 mmol) , intermediate 4 (160 mg, 0.53 mmol) and HATU (204 mg, 0.53 mmol) in DMF (5 mL) was added DIEA (140 mg, 1.1 mmol) . The reaction mixture was stirred at 25 °C for 2 h. The reaction mixture was poured into water (30 mL) . The aqueous phase was extracted with EtOAc (20 mL x 3) . The combined organic phase was washed with brine (30 mL x 2) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel to afford compound 146.4 (100 mg) as yellow oil. Chemical Formula: C 37 H 50 N 6 O 7 . Molecular Weight: LCMS (ESI) m/z: 691.4 [M+1] + .
  • Step 4 To a solution of compound 146.4 (100 mg, 1.0 eq) in DCM (2 mL) was added HCl/1, 4-dioxane (4 M, 2 mL) . The mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure to afford compound 146.5 (92 mg, crude) as a yellow solid. Chemical Formula: C 32 H 42 N 6 O 7 . Molecular Weight: 590.73. LCMS (ESI) m/z: 591.3 [M+1] + .
  • Step 5 To a solution of intermediate 3 (80 mg, 0.13 mmol) in MeOH (4 mL) was added NaOAc (25 mg, 0.30 mmol) . The mixture was stirred at 25°C for 10 min. Then the mixture was added compound 146.5 (92 mg, 0.16 mmol) , NaBH 3 CN (40 mg, 0.63 mmol) . The mixture was stirred at 25 °C for 16 h. The reaction mixture was poured into water (20 mL) . The aqueous phase was extracted with EtOAc (20 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure.
  • Step 6 To a solution of compound 146.6 (25 mg, 0.021 mmol) in DCM (2 mL) was added HCl/1, 4-dioxane (4 M, 2 mL) . The reaction mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (FA) to afford Example 146 (8.7 mg, 99%HPLC purity, 0.7 FA salt) as a white solid. Chemical Formula: C 60 H 68 F 2 N 10 O 7 . Molecular Weight: 1079.26. LCMS (ESI) m/z: 540.3 [1/2M+1] + .
  • Step 1 To a solution of intermediate 2 (200 mg, 1.0 mmol) , compound 149.1 (291 mg, 1.2 mmol) and PPh 3 (316 mg, 1.2 mmol) in Toluene (10 mL) was added DIAD (244.4 mg, 1.2 mmol) at 0 °C under N 2 . The reaction mixture was stirred at 100 °C for 16 h. The reaction mixture was poured into water (30 mL) . The aqueous phase was extracted with EtOAc (30 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure.
  • Step 2 To a solution of compound 149.2 (240 mg, 0.56 mmol) in THF/H 2 O/MeOH (4/2/2 mL) was added LiOH (40.9 mg, 1.6 mmol) . The reaction mixture was stirred at 25 °C for 2 h. The pH of the reaction mixture was adjusted to 5 with aq. HCl (1 M) . The aqueous phase was extracted with EtOAc (10 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure to afford compound 149.3 (245 mg) as a white solid. Chemical Formula: C 21 H 32 N 2 O 6 . Molecular Weight: 408.50. LCMS (ESI) m/z: 353.2 [M-56+1] + .
  • Step 3 To a solution of compound 149.3 (245 mg, 0.60 mmol) , intermediate 4 (268.6 mg, 0.90 mmol) and HATU (342.2 mg, 0.90 mmol) in DMF (10 mL) was added DIEA (234.5 mg, 1.8 mmol) . The reaction mixture was stirred at 25 °C for 2 h. The reaction mixture was poured into water (30 mL) . The aqueous phase was extracted with EtOAc (40 mL x 3) . The combined organic phase was washed with brine (50 mL x 2) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure.
  • Step 4 To a solution of compound 149.4 (360 mg, 1.0 eq) in DCM (5 mL) was added HCl/dioxane (4 M, 2 mL) . The mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure to afford compound 149.5 (360 mg, crude) as a yellow solid. Chemical Formula: C 33 H 44 N 6 O 5 . Molecular Weight: 604.75. LCMS (ESI) m/z: 605.3 [M+1] + .
  • Step 5 To a solution of intermediate 3 (50 mg, 0.082 mmol) in MeOH (4 mL) was added NaOAc (13.5 mg, 0.16 mmol) . The mixture was stirred at 25 °C for 10 min. Then the mixture was added compound 149.5 (55 mg, 0.090 mmol) , NaBH 3 CN (20.8 mg, 0.32 mmol) . The mixture was stirred at 25 °C for 16 h. The reaction mixture was poured into water (20 mL) . The aqueous phase was extracted with EtOAc (20 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure.
  • Step 6 To a solution of compound 149.6 (32 mg, 0.026 mmol) in DCM (5 mL) was added HCl/1, 4-dioxane (4 M, 2 mL) . The reaction mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (FA) to afford Example 149 (17.2 mg, 99%HPLC purity, 1.0 FA salt) as a white solid. Chemical Formula: C 61 H 70 F 2 N 10 O 7 . Molecular Weight: 1093.29. LCMS (ESI) m/z: 1094.4 [M+1] + .
  • Step 1 To a solution of intermediate 2 (200 mg, 0.91 mmol) , compound 150.1 (264 mg, 1.1 mmol) and PPh 3 (318 mg, 1.1 mmol) in anhydrous Toluene (10 mL) was added DIAD (244.4 mg, 1.1 mmol) at 0 °C under N 2 . The reaction mixture was stirred at 100 °C for 16 h. The reaction mixture was poured into water (30 mL) . The aqueous phase was extracted with EtOAc (30 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure.
  • Step 2 To a solution of compound 150.2 (280 mg, 0.7 mmol) in THF/H 2 O/MeOH (4/2/2 mL) was added LiOH (51 mg, 2.1 mmol) . The reaction mixture was stirred at 25 °C for 2 h. The pH of the reaction mixture was adjusted to 5 with aq. HCl (1 M) . The aqueous phase was extracted with EtOAc (10 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure to afford compound 150.3 (275 mg) as a white solid. Chemical Formula: C 18 H 27 FN 2 O 6 . Molecular Weight: 386.42. LCMS (ESI) m/z: 331.1 [M-56+1] + .
  • Step 3 To a solution of compound 150.3 (100 mg, 0.26 mmol) , intermediate 4 (116.2 mg, 0.38 mmol) and HATU (147.8 mg, 0.38 mmol) in DMF (10 mL) was added DIEA (100 mg, 0.77 mmol) . The reaction mixture was stirred at 25 °C for 2 h. The reaction mixture was poured into water (30 mL) . The aqueous phase was extracted with EtOAc (40 mL x 3) . The combined organic phase was washed with brine (50 mL x 2) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure.
  • Step 4 To a solution of compound 150.4 (360 mg, 0.52 mmol) in DCM (5 mL) was added HCl/1, 4-dioxane (4 M, 2 mL) . The mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure to afford compound 150.5 (360 mg, crude) as a yellow solid. Chemical Formula: C 30 H 39 FN 6 O 5 . Molecular Weight: 582.68. LCMS (ESI) m/z: 583.3 [M+1] + .
  • Step 5 To a solution of intermediate 3 (40 mg, 0.066 mmol) in MeOH (4 mL) was added NaOAc (10.8 mg, 0.13 mmol) . The mixture was stirred at 25 °C for 10 min. Then compound 150.5 (46.3 mg, 0.072 mmol) and NaBH 3 CN (16.8 mg, 0.26 mmol) were added. The mixture was stirred at 25 °C for 16 h. The reaction mixture was poured into water (20 mL) . The aqueous phase was extracted with EtOAc (20 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure.
  • Step 6 To a solution of compound 150.6 (55 mg, 0.047 mmol) in DCM (5 mL) was added HCl/1, 4-dioxane (4 M, 2 mL) . The reaction mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (FA) to afford Example 150 (35.5 mg, 99%HPLC purity, 0.8 FA salt) as a white solid. Chemical Formula: C 58 H 65 F 3 N 10 O 7 . Molecular Weight: 1071.22. LCMS (ESI) m/z: 1072.3 [M+1] + .
  • Step 1 To a solution of intermediate 2 (202 mg, 1.0 mmol) , compound 151.1 (250 mg, 1.2 mmol) and PPh 3 (320 mg, 1.2 mmol) in anhydrous toluene (5 mL) was added DIAD (250 mg, 1.2 mmol) at 90 °C under N 2 . The reaction mixture was stirred at 90 °C for 8 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel to afford compound 151.2 (220 mg) as yellow oil. Chemical Formula: C 18 H 27 FN 2 O 6 . Molecular Weight: 386.42. LCMS (ESI) m/z: 331.1 [M-56+1] + .
  • Step 2 To a solution of compound 151.2 (220 mg, 0.57 mmol) in THF/H 2 O/MeOH (2/2/2 mL) was added LiOH (36 mg, 0.85 mmol) . The reaction mixture was stirred at 25 °C for 2 h. The pH of the reaction mixture was adjusted to 5 with aq. HCl (1 M) . The aqueous phase was extracted with EtOAc (10 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure to afford compound 151.3 (200 mg) as yellow oil. Chemical Formula: C 17 H 25 FN 2 O 6 . Molecular Weight: 372.39. LCMS (ESI) m/z: 317.1 [M-56+1] + .
  • Step 3 To a solution of compound 151.3 (100 mg, 0.27 mmol) , intermediate 4 (120 mg, 0.40 mmol) and HATU (153 mg, 0.40 mmol) in DMF (5 mL) was added DIEA (104 mg, 0.80 mmol) . The reaction mixture was stirred at 25 °C for 2 h. The reaction mixture was poured into water (30 mL) . The aqueous phase was extracted with EtOAc (20 mL x 3) . The combined organic phase was washed with brine (30 mL x 2) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel to afford compound 151.4 (100 mg) as yellow oil. Chemical Formula: C 34 H 45 FN 6 O 7 . Molecular Weight: 668.77. LCMS (ESI) m/z: 669.7 [M+1] + .
  • Step 4 To a solution of compound 151.4 (100 mg, 0.15 mmol) in DCM (2 mL) was added HCl/1, 4-dioxane (4 M, 2 mL) . The mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure to afford compound 151.5 (100 mg, crude) as a yellow solid. Chemical Formula: C 29 H 37 FN 6 O 7 . Molecular Weight: 568.68. LCMS (ESI) m/z: 569.3 [M+1] + .
  • Step 5 To a solution of intermediate 3 (40 mg, 0.066 mmol) in MeOH (4 mL) was added NaOAc (12 mg, 0.15 mmol) . The mixture was stirred at 25 °C for 10 min. Then compound 151.5 (50 mg, 0.088 mmol) and NaBH 3 CN (19 mg, 0.30 mmol) were added. The mixture was stirred at 25 °C for 16 h. The reaction mixture was poured into water (20 mL) . The aqueous phase was extracted with EtOAc (20 mL x 2) . The combined organic phase was washed with brine (30 mL) and dried over Na 2 SO 4 . The organic layer was concentrated under reduced pressure.
  • Step 6 To a solution of compound 151.6 (30 mg, 0.026 mmol) in DCM (2 mL) was added HCl/dioxane (4 M, 2 mL) . The reaction mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (FA) to Example 151 (18.6 mg, 99%HPLC purity, 0.5 FA salt) as a white solid. Chemical Formula: C 57 H 63 F 3 N 10 O 7 . Molecular Weight: 1057.19. LCMS (ESI) m/z: 529.3 [1/2 M+1] + .
  • All cancer cell lines were obtained from American Type Culture Collection (ATCC, USA) .
  • Pancreatic cancer cell lines SW1990 and AsPC-1 cells both harbor homozygous KRAS G12D mutation.
  • GP2D harboring heterozygous KRAS G12D mutation, is derived from colon cancer.
  • HCT116 is colon cancer cell line harboring KRAS G13D mutation.
  • Mia-PaCa-2 is pancreatic cancer cell with KRAS G12C mutation. Compounds having pharmacological effect on these cells have implication extending to all other cancer cells containing such KRAS mutations.
  • Cancer cell lines HCT116 carrying KRAS G13D mutation, AsPC-1 carrying homozygous KRAS G12D mutation, and Mia-PaCa-2 carrying homozygous KRAS G12C mutation were seeded in 6-well plate, incubated overnight at 37°C and 5%CO2.
  • AsPC-1 cells were seeded into 384-well round bottom plate and chemical compounds for growth inhibition were added at 1 uM, 333 nM, 111 nM, 37 nM, 12 nM, 4 nM, 1 nM, 0.5 nM, 0.2 nM, DMSO (vehicle solvent) .
  • the IC50 of the tested compounds were derived from fitting non-linear regression equation:
  • Top and Bottom Plateaus in same units as Y.
  • logIC50 same log units as X.
  • HillSlope Slope factor or Hill slope.
  • A means ⁇ 100 nM
  • B means ⁇ 100 nM and ⁇ 1000 nM
  • C means ⁇ 1000 nM
  • N means no data available
  • A means ⁇ 100 nM
  • B means ⁇ 100 nM and ⁇ 1000 nM
  • C means ⁇ 1000 nM
  • N.D. means no data available

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Abstract

L'invention concerne des composés bifonctionnels, qui sont utiles en tant que modulateurs de la protéine Kirsten ras sarcoma (KRas ou KRAS). La présente invention concerne particulièrement, des composés hétéro-bifonctionnels contenant, à une extrémité une fraction qui se lie à l'ubiquitine ligase E3 de Von Hippel-Lindau, et à l'autre extrémité, une fraction qui se lie à KRas, de telle sorte que la protéine cible est placée à proximité de l'ubiquitine ligase pour effectuer une dégradation (et une inhibition) de la protéine cible. Les composés hétéro-bifonctionnels selon la présente divulgation présentent une large plage d'activités pharmacologiques associées à la dégradation/l'inhibition de protéine cible. Les composés et les compositions selon la présente divulgation permettent de traiter ou de prévenir des maladies ou des troubles consécutifs à une régulation aberrante de la protéine cible.
PCT/CN2023/084424 2022-03-28 2023-03-28 Composés pour la dégradation ciblée de kras WO2023185864A1 (fr)

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Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112218859A (zh) * 2018-04-04 2021-01-12 阿尔维纳斯运营股份有限公司 蛋白水解调节剂及相关使用方法
WO2021207172A1 (fr) * 2020-04-06 2021-10-14 Arvinas Operations, Inc. Composés et procédés de dégradation ciblée de kras
WO2022148422A1 (fr) * 2021-01-08 2022-07-14 Beigene, Ltd. Composés pontés en tant qu'inhibiteur et dégradeur de kras g12d et leur utilisation
WO2022266206A1 (fr) * 2021-06-16 2022-12-22 Erasca, Inc. Conjugués d'inhibiteurs de kras
WO2023280026A1 (fr) * 2021-07-05 2023-01-12 四川科伦博泰生物医药股份有限公司 Composé cyclique hétéroaromatique, son procédé de préparation et son utilisation
WO2023077441A1 (fr) * 2021-11-05 2023-05-11 Ranok Therapeutics (Hangzhou) Co. Ltd. Procédés et compositions pour la dégradation ciblée de protéines

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112218859A (zh) * 2018-04-04 2021-01-12 阿尔维纳斯运营股份有限公司 蛋白水解调节剂及相关使用方法
WO2021207172A1 (fr) * 2020-04-06 2021-10-14 Arvinas Operations, Inc. Composés et procédés de dégradation ciblée de kras
WO2022148422A1 (fr) * 2021-01-08 2022-07-14 Beigene, Ltd. Composés pontés en tant qu'inhibiteur et dégradeur de kras g12d et leur utilisation
WO2022266206A1 (fr) * 2021-06-16 2022-12-22 Erasca, Inc. Conjugués d'inhibiteurs de kras
WO2023280026A1 (fr) * 2021-07-05 2023-01-12 四川科伦博泰生物医药股份有限公司 Composé cyclique hétéroaromatique, son procédé de préparation et son utilisation
WO2023077441A1 (fr) * 2021-11-05 2023-05-11 Ranok Therapeutics (Hangzhou) Co. Ltd. Procédés et compositions pour la dégradation ciblée de protéines

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