WO2026013576A1 - Dérivés de dihydrouracile utiles pour la dégradation ciblée de vav1 - Google Patents

Dérivés de dihydrouracile utiles pour la dégradation ciblée de vav1

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Publication number
WO2026013576A1
WO2026013576A1 PCT/IB2025/056919 IB2025056919W WO2026013576A1 WO 2026013576 A1 WO2026013576 A1 WO 2026013576A1 IB 2025056919 W IB2025056919 W IB 2025056919W WO 2026013576 A1 WO2026013576 A1 WO 2026013576A1
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group
compound
independently selected
pharmaceutically acceptable
ring
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Inventor
Xavier Lucas CABRÉ
Bernhard FASCHING
Elisa Liardo
Laura Ann Mcallister
Vladimiras OLEINIKOVAS
Andreas RITZÉN
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Monte Rosa Therapeutics AG
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Monte Rosa Therapeutics AG
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/10—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a carbon chain containing aromatic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • 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
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/10—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a carbon chain containing aromatic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
    • C07D413/10—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a carbon chain containing aromatic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/10—Spiro-condensed systems
    • C07D491/107—Spiro-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring

Definitions

  • VAV1 Proto-oncogene VAV 1 protein
  • VAV1 Proto-oncogene VAV 1 protein
  • This disclosure also features compositions containing these chemical entities as well as methods of using and making these chemical entities. 10 BACKGROUND The ubiquitin proteasome system can be manipulated with different small molecules to trigger targeted degradation of specific proteins of interest.
  • E3 ligases such as cereblon (a phenomenon known as E3 reprogramming) using low molecular weight compounds, which have been termed molecular glues (also called molecular glue degraders; “MGDs”), to promote the poly-ubiquitination and ultimately proteasomal degradation of new protein substrates involved in the development of diseases.
  • MLDs molecular glue degraders
  • molecular glues for the E3 ligase cereblon include: Thalidomide, Lenalidomide and Pomalidomide, all of which are immunomodulatory imide drugs (IMiDs) approved by the FDA for use in hematological cancers.
  • IMDs immunomodulatory imide drugs
  • VAV family proteins, including VAV1, VAV2 and VAV3, are guanine nucleotide exchange factors (GEFs) for Rho family GTPases.
  • VAV1 is a 95 kDa protein that is a positive regulator of T cell receptor and B cell receptor signaling.
  • VAV1 expression is normally highly restricted to hematopoietic cells. VAV1 becomes rapidly phosphorylated on tyrosine in response to a variety of stimuli, including stimulation of T-cell receptor (TCR), B cell receptor (BCR), and 30 various cytokine receptors. VAV1 regulates multiple cellular functions and signaling pathways in hematopoietic-derived cells (e.g., T- and B-cells, natural killer cells, and osteoclasts) through activation of certain GTPases. VAV1-mediated functions include gene transcription, development and activation of immune cells (e.g., T- and B-cells). VAV1 is a positive regulator of (TCR) 1
  • PAT059977-WO-PCT signaling including nuclear factor of activated T cells (NFAT), interferon gamma (IFN ⁇ ) and Interleukin-2 (IL-2) cytokine secretion.
  • NFAT nuclear factor of activated T cells
  • IFN ⁇ interferon gamma
  • IL-2 Interleukin-2
  • VAV1 deficient mice are resistant to MOG(5-55)-induced experimental autoimmune encephalomyelitis (EAE), a commonly used model of multiple sclerosis (Korn et al. 2003 Journal of Neuroimmunology 139:17).
  • EAE experimental autoimmune encephalomyelitis
  • CRISPRa genome-wide CRISPR activation
  • CRISPRi interference
  • SUMMARY This disclosure features chemical entities (e.g., a compound or a pharmaceutically 15 acceptable salt thereof) that degrade Proto-oncogene VAV 1 protein (VAV1).
  • these chemical entities are useful, e.g., for treating a subject (e.g., a human subject) having a disorder or disease that can be treated by reducing the level of VAV1, thereby reducing VAV activity in cells.
  • the chemical entities can reduce signaling in certain immune cell activation pathways.
  • the chemical entities may be used to reduce inflammation or 20 autoimmune activity. They may be useful for treating, for example, multiple sclerosis, rheumatoid arthritis, myasthenia gravis, chronic lymphocytic leukemia, ulcerative colitis, psoriasis, cutaneous lupus, axial spondylarthritis graft versus host disease, and other disorders referred to herein.
  • VAV1 is a dominant signal transduction protein in the adaptive immune system. It is a positive regulator of immune receptor signaling in both T cells and B cells. Thus, reduction in VAV1 can reduce immune cell activation, immune cell proliferation and the production of various cytokines. For at least these reasons, degradation of VAV1 can be therapeutically beneficial in a variety of disease conditions.
  • this disclosure features compounds of Formula (I) or pharmaceutically acceptable salts thereof, 2
  • pharmaceutically acceptable salt refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate 10 with a reasonable benefit/risk ratio.
  • Pharmaceutically acceptable salts are well known in the art. For example, Berge et al., describes pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences (1977) 66:1–19.
  • Pharmaceutically acceptable salts of the compounds of the present disclosure include those derived from suitable inorganic and organic acids and bases.
  • suitable inorganic and organic acids and bases include those derived from suitable inorganic and organic acids and bases.
  • pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group 15 formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange.
  • salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, 20 citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2–hydroxy–ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2–naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pec
  • Pharmaceutically acceptable salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N + (C 1–4 alkyl) 4 salts.
  • Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like.
  • Further pharmaceutically acceptable salts include, when appropriate, nontoxic 3
  • VAV1 refers to naturally occurring VAV1, also known as Vav or p95vav, (e.g. mammalian, preferably human (Homo sapiens) VAV1) and encompasses naturally 5 occurring variants, such as allelic variants and splice variants, which retain VAV1 functional activity.
  • a “subject” to which administration is contemplated includes, but is not limited to, humans (i.e., a male or female of any age group, e.g., a pediatric subject (e.g, infant, child, adolescent) or adult subject (e.g., young adult, middle–aged adult or senior adult)) and/or a non-human animal, 10 e.g., a mammal such as primates (e.g., cynomolgus monkeys, rhesus monkeys), cattle, pigs, horses, sheep, goats, rodents, cats, and/or dogs.
  • the subject is a human.
  • the subject is a non-human animal.
  • the subject is a human.
  • Disease, disorder, and condition are used interchangeably herein.
  • the terms “treat,” “treating” and 15 “treatment” contemplate an action that occurs while a subject is suffering from the specified disease, disorder or condition, which reduces the severity of the disease, disorder or condition, or retards or slows the progression of the disease, disorder or condition (“therapeutic treatment”).
  • therapeutic treatment contemplate an action that occurs while a subject is suffering from the specified disease, disorder or condition, which reduces the severity of the disease, disorder or condition, or retards or slows the progression of the disease, disorder or condition (“therapeutic treatment”).
  • the “effective amount” of a compound refers to an amount sufficient to elicit the desired biological response.
  • the 20 effective amount of a compound of the present disclosure may vary depending on such factors as the desired biological endpoint, the pharmacokinetics of the compound, the disease being treated, the mode of administration, and the age, health, and condition of the subject.
  • a “therapeutically effective amount” of a compound is an amount sufficient to provide a therapeutic benefit in the treatment of a disease, 25 disorder or condition, or to delay or minimize one or more symptoms associated with the disease, disorder or condition.
  • a therapeutically effective amount of a compound means an amount of therapeutic agent, alone or in combination with other therapies, which provides a therapeutic benefit in the treatment of the disease, disorder or condition.
  • the term “therapeutically effective amount” can encompass an amount that improves overall therapy, reduces or avoids symptoms or 30 causes of disease or condition, or enhances the therapeutic efficacy of another therapeutic agent.
  • the compounds described herein also include isotopically labeled compounds which are identical to those recited herein, except that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds described herein include 35 isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine, such as 4
  • a compound of the disclosure may have one or more H atom replaced with deuterium.
  • halo refers to fluoro (F), chloro (Cl), bromo (Br), or iodo (I).
  • alkyl refers to a saturated acyclic hydrocarbon radical that may be a straight 5 chain or branched chain, containing the indicated number of carbon atoms. For example, C1-10 indicates that the group may have from 1 to 10 (inclusive) carbon atoms in it.
  • Non-limiting examples include methyl, ethyl, iso-propyl, tert-butyl, n-hexyl.
  • saturated as used in this context means only single bonds present between constituent carbon atoms and other available valences occupied by hydrogen and/or other substituents as defined herein. 10
  • haloalkyl refers to an alkyl, in which one or more hydrogen atoms is/are replaced with an independently selected halo.
  • alkoxy refers to an -O-alkyl radical (e.g., -OCH3).
  • alkylene refers to a divalent alkyl (e.g., -CH 2 -).
  • alkenyl refers to an acyclic hydrocarbon chain that may be a straight chain or 15 branched chain having one or more carbon-carbon double bonds.
  • the alkenyl moiety contains the indicated number of carbon atoms.
  • C 2-6 indicates that the group may have from 2 to 6 (inclusive) carbon atoms in it.
  • alkynyl refers to an acyclic hydrocarbon chain that may be a straight chain or branched chain having one or more carbon-carbon triple bonds.
  • the alkynyl moiety contains the 20 indicated number of carbon atoms.
  • C 2-6 indicates that the group may have from 2 to 6 (inclusive) carbon atoms in it.
  • aryl refers to a 6-20 carbon mono-, bi-, tri- or polycyclic group wherein at least one ring in the system is aromatic (e.g., 6-carbon monocyclic, 10-carbon bicyclic, or 14-carbon tricyclic aromatic ring system).
  • aryl groups include phenyl, naphthyl, 25 tetrahydronaphthyl, dihydro-1H-indenyl and the like.
  • cycloalkyl refers to cyclic saturated hydrocarbon groups having, e.g., 3 to 20 ring carbons, preferably 3 to 16 ring carbons, and more preferably 3 to 12 ring carbons or 3-10 ring carbons or 3-6 ring carbons.
  • Cycloalkyl groups include, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.
  • Cycloalkyl may include multiple fused and/or bridged rings.
  • Non-limiting examples of fused/bridged cycloalkyl includes: bicyclo[1.1.0]butanyl, bicyclo[2.1.0]pentanyl, bicyclo[1.1.1]pentanyl, bicyclo[3.1.0]hexanyl, bicyclo[2.1.1]hexanyl, bicyclo[3.2.0]heptanyl, bicyclo[4.1.0]heptanyl, bicyclo[2.2.1]heptanyl, bicyclo[3.1.1]heptanyl, bicyclo[4.2.0]octanyl, bicyclo[3.2.1]octanyl, bicyclo[2.2.2]octanyl, and the like. Cycloalkyl also includes spirocyclic 35 rings (e.g., spirocyclic bicycle wherein two rings are connected through just one atom).
  • PAT059977-WO-PCT limiting examples of spirocyclic cycloalkyls include spiro[2.2]pentanyl, spiro[2.5]octanyl, spiro[3.5]nonanyl, spiro[3.5]nonanyl, spiro[3.5]nonanyl, spiro[4.4]nonanyl, spiro[2.6]nonanyl, spiro[4.5]decanyl, spiro[3.6]decanyl, spiro[5.5]undecanyl, and the like.
  • saturated as used in this context means only single bonds present between constituent carbon atoms.
  • cycloalkenyl as used herein means partially unsaturated cyclic hydrocarbon groups having 3 to 20 ring carbons, preferably 3 to 16 ring carbons, and more preferably 3 to 12 ring carbons or 3-10 ring carbons or 3-6 ring carbons.
  • Examples of cycloalkenyl groups include, without limitation, cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl.
  • cycloalkenyl groups may have any degree of unsaturation 10 provided that one or more double bonds is present in the ring, none of the rings in the ring system are aromatic, and the cycloalkenyl group is not fully saturated overall.
  • Cycloalkenyl may include multiple fused and/or bridged and/or spirocyclic rings.
  • heteroaryl as used herein, means a mono-, bi-, tri- or polycyclic group having 5 to 10 ring atoms and having pi electrons shared in a cyclic array; wherein at least one ring in the 15 system is aromatic, and at least one ring in the system contains one or more heteroatoms independently selected from the group consisting of N, O, and S (but the aromatic ring does not have to be a ring which contains a heteroatom, e.g. tetrahydroisoquinolinyl, e.g., tetrahydroquinolinyl).
  • heteroaryl examples include thienyl, pyridinyl, furyl, oxazolyl, oxadiazolyl, pyrrolyl, imidazolyl, triazolyl, thiodiazolyl, pyrazolyl, isoxazolyl, thiadiazolyl, 20 pyranyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, thiazolyl benzothienyl, benzoxadiazolyl, benzofuranyl, benzimidazolyl, benzotriazolyl, cinnolinyl, indazolyl, indolyl, isoquinolinyl, isothiazolyl, naphthyridinyl, purinyl, thienopyridinyl, pyrido[2,3-d]pyrimidinyl, pyrrolo[2,3- b]pyridinyl, quinazolin
  • the heteroaryl is selected from thienyl, pyridinyl, furyl, pyrazolyl, imidazolyl, isoindolinyl, pyranyl, pyrazinyl, and pyrimidinyl.
  • heterocyclyl refers to a mono-, bi-, tri-, or polycyclic saturated ring system 30 having 5-10 ring atoms (e.g., a 4-6-membered monocyclic, or 7-9-membered bicyclic ring system), wherein one or more ring atoms are heteroatoms, said heteroatoms selected from O, N, or S.
  • heterocyclyl groups include piperazinyl, pyrrolidinyl, dioxanyl, morpholinyl, tetrahydrofuranyl, and the like.
  • Heterocyclyl may include multiple fused and bridged rings.
  • fused/bridged heteorocyclyl includes: 2-azabicyclo[1.1.0]butanyl, 2- 35 azabicyclo[2.1.0]pentanyl, 2-azabicyclo[1.1.1]pentanyl, 3-azabicyclo[3.1.0]hexanyl, 5- 6
  • Heterocyclyl also includes spirocyclic rings (e.g., spirocyclic bicycle wherein two rings are connected through just one atom).
  • spirocyclic heterocyclyls include 2-10 azaspiro[2.2]pentanyl, 4-azaspiro[2.5]octanyl, 1-azaspiro[3.5]nonanyl, 2-azaspiro[3.5]nonanyl, 7- azaspiro[3.5]nonanyl, 2-azaspiro[4.4]nonanyl, 6-azaspiro[2.6]nonanyl, 1,7- diazaspiro[4.5]decanyl, 7-azaspiro[4.5]decanyl 2,5-diazaspiro[3.6]decanyl, 3- azaspiro[5.5]undecanyl, 2-oxaspiro[2.2]pentanyl, 4-oxaspiro[2.5]octanyl, 1- oxaspiro[3.5
  • heterocycloalkenyl as used herein means partially unsaturated cyclic ring system with 5-10 ring atoms (e.g., a monocyclic bicyclic or tricyclic ring system), wherein one or more ring atoms are heteroatoms, said heteroatoms selected from O, N, or S.
  • heterocycloalkenyl groups include, without limitation, tetrahydropyridyl, dihydropyrazinyl, dihydropyridyl, dihydropyrrolyl, dihydrofuranyl, dihydrothiophenyl.
  • heterocycloalkenyl groups may have any degree of unsaturation provided that one or more double bonds is present in the ring, none of the rings in the ring system are aromatic, and the heterocycloalkenyl group is not fully saturated overall.
  • Heterocycloalkenyl may include multiple fused and/or bridged and/or spirocyclic rings.
  • Certain groups can be considered as either: (i) a heterocycloalkenyl 30 which is substituted with an oxo group; or (ii) a heteroaryl group.
  • a ring when a ring is described as being “aromatic”, it means said ring has a continuous, delocalized ⁇ -electron system. Typically, the number of out of plane ⁇ -electrons corresponds to the Hückel rule (4n+2). Examples of such rings include: benzene, pyridine, 7
  • PAT059977-WO-PCT pyrimidine pyrazine, pyridazine, pyridone, pyrrole, pyrazole, oxazole, thioazole, isoxazole, isothiazole, and the like.
  • a ring when a ring is described as being “partially unsaturated”, it means said ring has one or more additional degrees of unsaturation (in addition to the degree of unsaturation 5 attributed to the ring itself; e.g., one or more double or triple bonds between constituent ring atoms), provided that the ring is not aromatic.
  • rings examples include: cyclopentene, cyclohexene, cycloheptene, dihydropyridine, tetrahydropyridine, dihydropyrrole, dihydrofuran, dihydrothiophene, and the like.
  • rings and cyclic groups 10 e.g., aryl, heteroaryl, heterocyclyl, heterocycloalkenyl, cycloalkenyl, cycloalkyl, and the like described herein
  • rings and cyclic groups encompass those having fused rings, including those in which the points of fusion are located (i) on adjacent ring atoms (e.g., [x.x.0] ring systems, in which 0 represents a zero atom 15 , ystems having
  • atoms making up the compounds of the present embodiments are intended to 20 include all isotopic forms of such atoms.
  • Isotopes include those atoms having the same atomic number but different mass numbers.
  • isotopes of hydrogen include tritium and deuterium
  • isotopes of carbon include 13 C and 14 C.
  • the compounds generically or specifically disclosed herein include all 25 tautomeric forms, or “tautomers” of said compounds.
  • PAT059977-WO-PCT of a compound with the group is also a disclosure of a tautomer of that compound, for instance with the group .
  • the compounds generically or specifically disclosed herein include all stereoisomeric forms, including all diastereomeric and entantiomeric forms, unless it is specifically stated or the 5 context indicates otherwise.
  • Compounds with chiral centers can occur as racemates, individual enantiomers (e.g. as the (R) enantiomer or (S) entantiomer) or diastereomers, and mixtures thereof. All such stereoisomeric forms are included within the embodiments disclosed herein, including mixtures thereof.
  • a compound comprising a chiral center disclosed herein without its enantiomeric 10 form indicated encompasses the isolated entantiomer and a mixture, such as a racemic mixture, of the (R) and (S) entantiomers if the enantiomers epimerise.
  • the phrase “optionally substituted” when used in conjunction with a structural moiety is intended to encompass both the unsubstituted structural moiety (i.e., none of the substitutable hydrogen atoms are replaced with one or more non-hydrogen15 substituents) and substituted structural moieties substituted with the indicated range of non- hydrogen substituents.
  • C1-C4 alkyl optionally substituted with 1-4 R a is intended to encompass both unsubstituted C 1 -C 4 alkyl and C 1 -C 4 alkyl substituted with 1-4 R a .
  • the term “antibody” encompasses an immunoglobulin, whether natural or partly or wholly synthetically produced, and fragments thereof. The term also covers any protein 20 having a binding domain that is homologous to an immunoglobulin binding domain. “Antibody” further includes a polypeptide comprising a framework region from an immunoglobulin gene or fragments thereof that specifically binds and recognizes an antigen.
  • FIG. 1 shows compound VAV1 MGD concentration in serum (left) after seven consecutive daily doses at increasing doses and associated VAV1 levels normalized to b-actin protein levels in PBMCs relative to vehicle 6 and 24 hours after 7 consecutive daily oral doses of VAV1 MGD at indicated doses (right).
  • FIG.2 shows dose-dependent decrease in VAV1 levels in primary human CD3+ T cells, 20 CD19+ B cells, and CD14+ monocytes following 24h VAV1 MGD treatment relative to DMSO control as assessed by flow cytometry (y-axis represents normalized VAV1 levels to DMSO control; x-axis depicts doses of VAV1 MGD).
  • FIG. 3 shows that VAV1 degradation results in inhibition of various hallmarks of TCR- mediated activity following TCR stimulation of primary human T-cells. Cells were treated with 25 VAV1 MGD for 24h followed by TCR stimulation (anti-CD3/anti-CD28).
  • FIG. 4 shows a schematic diagram of the role of VAV1 in B cell receptor signaling. Following the ligation of the B cell receptor and/or CD19, VAV1 is recruited to mediate phosphorylation cascades that lead to activation, cytokine secretion, and antibody production by B cells. 10
  • FIG. 5 shows a schematic diagram of the role of VAV1 in T cell receptor signaling. Following the ligation of the T cell receptor, VAV1 is recruited to mediate phosphorylation cascades that lead to activation, cytokine secretion, and proliferation.
  • FIG. 6 shows that VAV1 MGD-mediated degradation of VAV1 reduces BCR-mediated 5 CD69 expression and secretion of IL-6 and IgG of primary human B cells.
  • Purified human primary B-cells were treated with MGD for 24 hrs followed by stimulation with anti-IgM and recombinant human IL-4 for 24 hours (for CD69 expression and IL-6 secretion) or with anti-IgM, BAFF, IL- 21, and sCD40L for 5 days (for IgG secretion).
  • CD69 expression was then assessed on CD19+ B cells by flow cytometry.
  • CD69 expression is shown as a percentage (%) change relative to 10 stimulated DMSO controls.
  • X-axis shows relative percentage of CD19+ B cells expressing CD69 and y-axis shows concentration of VAV1 MGD.
  • IL-6 secretion was assessed in the supernatant by Alphalisa.
  • IL-6 secretion is shown as a percentage (%) change relative to stimulated DMSO controls.
  • X-axis shows relative percentage of IL-6 level and y-axis shows concentration of VAV1 MGD.
  • IgG secretion was assessed in the supernatant by Alphalisa.
  • IgG secretion is shown as a 15 percentage (%) change relative to stimulated DMSO controls.
  • X-axis shows relative percentage of IgG level and y-axis shows concentration of VAV1 MGD.
  • FIG.7 shows that VAV1 degradation results in inhibition TNF secretion following Fc ⁇ R stimulation of primary human monocytes. Cells were treated with VAV1 MGD for 24h followed by FC ⁇ R stimulation (200 ⁇ g/mL immobilized IgG).
  • FIG.8 shows that VAV1 MGD treatment of REC-1 cell line degraded VAV1 in a concentration-dependent manner. REC-1 were treated for 24 hours with the indicated concentrations of VAV1 MGD. After treatment, VAV1 levels were assessed by western blot. 25 Data shows VAV1 levels normalized to ⁇ -actin and relative to DMSO control.
  • FIG.9 shows that VAV1 MGD treatment of REC-1 cell line decreases growth with increasing concentration. REC-1 were treated for 5 days with the indicated concentrations of VAV1 MGD.
  • FIG.10A shows that oral dosing of VAV1 MGDs inhibits disease progression in a mouse model of EAE.
  • EAE was induced in C57BL/6J mice and mice were treated orally and daily from Day 11 with either vehicle (10% captisol in water) or Compound 1 at 10 mg/kg or Compound 24 at 30, 10, and 1 mg/kg. Mice were assessed for disease severity every 3 days until 11
  • FIG.10B shows that oral dosing of peripherally-restricted VAV1 MGDs degrades VAV1 in peripheral blood mononuclear cells (PBMC).
  • PBMCs peripheral blood mononuclear cells
  • Image shows representative VAV1 levels in each treatment group as indicated. Quantification shows relative levels in VAV1 normalized to vehicle-treated group for all mice. Bars indicate mean ⁇ SEM.
  • FIG.10C shows that oral dosing of peripherally-restricted VAV1 MGDs does not 10 degrade VAV1 in brain tissue.
  • brain tissue was excised, homogenized, and assessed for VAV1 and ⁇ -actin levels by western blot.
  • Image shows representative VAV1 levels in each treatment group as indicated.
  • Quantification shows relative levels in VAV1 normalized to vehicle-treated group for all mice. Bars indicate mean ⁇ SEM. 15 DETAILED DESCRIPTION
  • This disclosure features chemical entities (e.g., a compound or a pharmaceutically acceptable salt thereof) that degrade and/or otherwise inhibit Proto-oncogene VAV 1 protein (VAV1).
  • Said chemical entities are useful, e.g., for treating a subject (e.g., a human subject) having a disorder or disease associated with VAV1 polymorphisms.
  • This disclosure also features 20 compositions containing the same as well as methods of using and making the same.
  • Compounds This disclosure features compounds of Formula (I) or pharmaceutically acceptable salts thereof, 25 Formula (I) wherein: Ring A is selected from the group consisting of: 12
  • each occurrence of R 14 is independently selected from the group consisting of: deuterium; -OH; -halo; C1-2 alkyl; C1-2 alkoxy; C1-2 haloalkyl; and C1-2 haloalkoxy.
  • the compound has Formula (II): 5 wherein X 1 , X 2 and X 3 are each selected from the group consisting of CR 6D and N, and 10 wherein a maximum of one of X 1 , X 2 and X 3 may be N.
  • the compound has Formula (III): 15 wherein Y and Z are independently selected from the group consisting of CH, CR 14 and N, wherein at least one of Y and Z is N and wherein m is 0 or 1. In some embodiments, the compound has Formula (IV): 20 Formula (IV) wherein Y is selected from the group consisting of CH, CR 14 and N. 15
  • R 2 is Cl.
  • at least one of R 3 , R 4 and R 5 is H.
  • at least two of 5 R 3 , R 4 and R 5 is H.
  • all of R 3 , R 4 and R 5 are H.
  • R 9 is hydrogen.
  • the compound is of Formula (V): 10 Formula (V) or a pharmaceutically acceptable salt thereof.
  • Ring A is selected from the group consisting of: 15 • , wherein * is the point of attachment to L, X 1 , X 2 and X 3 are each selected from the group consisting of CH and N, and wherein a maximum of one of X 1 , X 2 and X 3 may be N; or • , wherein Z is the point of attachment to L, and Y and Z are independently selected from the group consisting of CH and N; 20
  • Ring B is selected from the group consisting of: • heterocyclyl or heterocycloalkenyl including 5-9 ring atoms, wherein from 1-4 ring atoms are heteroatoms, each independently selected from the group consisting of N, N(H), N(R 1 ) and O, and wherein one or more of the carbon atoms of the heterocyclyl or heterocycloalkenyl are optionally substituted with from 1-4 substituents independently 25 selected from the group consisting of oxo and R 8 ; or • heteroaryl including 5-9 ring atom
  • PAT059977-WO-PCT least one ring in the system is aromatic and wherein one or more of the carbon atoms of the heteroaryl are optionally substituted with from 1-4 substituents independently selected from oxo and R 8 ; and L is selected from the group consisting of a bond, *-O(C1-C4 alkylene)-, and *-C1-C4 alkylene-, 5 wherein the alkylene is optionally substituted with 1-2 R 7 , wherein * denotes the point of attachment of L to Ring A, wherein L is a bond or *-C 1 -C 4 alkylene- when Z is N.
  • Ring A is selected from the group consisting of: • , wherein * is the point of attachment to L, wherein X 1 and X 2 10 are each selected from the group consisting of CH and N and wherein a maximum of one of X 1 and X 2 may be N; or • , wherein N is the point of attachment to L, and wherein Y is selected from the group consisting of CH and N;
  • Ring B is selected from the group consisting of: 15 • heterocyclyl or heterocycloalkenyl including 5-9 ring atoms, wherein from 1-3 ring atoms are heteroatoms, each independently selected from the group consisting of N, N(H), N(R 1 ) and O, wherein one or more of the carbon atoms of the heterocyclyl or heterocycloalkenyl is optionally substituted with from 1-4 substituents independently selected from the group consisting of oxo and R 8 ; or 20 • heteroaryl including 5-6 ring atoms, wherein from 1-2 ring atoms,
  • PAT059977-WO-PCT • wherein * is the point of attachment to L, X 1 , X 2 and X 3 are each selected from the group consisting of CH and N, and wherein a maximum of one of X 1 , X 2 and X 3 may be N; or • , wherein Z is the point of attachment to L and Y and Z are 5 independently selected from the group consisting of CH and N.
  • Ring A is selected from the group consisting of: • , wherein * is the point of attachment to L, wherein X 1 and X 2 are each selected from the group consisting of CH and N and wherein a maximum 10 of one of X 1 and X 2 may be N.
  • Ring , wherein * is the point of attachment to L, X 1 , X 2 and X 3 are each selected from the group consisting of CH and N, and wherein a maximum of 15 one of X 1 , X 2 and X 3 may be N.
  • Ring , wherein * is the point of attachment to L, X 1 and X 2 are each selected from the group consisting of CH and N, and wherein a maximum of one of X 1 and X 2 may be N.
  • Ring wherein * is the point of attachment to L, and wherein X 1 is selected from the group consisting of CH and N. In some embodiments, Ring , wherein * is the point of attachment to L. In some embodiments, Ring . 5 In some embodiments, Ring , wherein Z is the point of attachment to L, Y and Z are independently selected from the group consisting of CH and N. In some embodiments, Ring .
  • Ring 10 is selected from the group consisting of: ,
  • L is selected from the group consisting of a bond, *-O(C 1 -C 4 alkylene)-, 15 and *-C 1 -C 4 alkylene-, wherein the alkylene is optionally substituted with 1-2 R 7 , wherein * 19
  • PAT059977-WO-PCT denotes the point of attachment of L to Ring A, wherein L is a bond or *-C1-C4 alkylene- when Z is N.
  • L is selected from the group consisting of a bond, *-methylene and *-O- 5 methylene, wherein * indicates the point of attachment of L to Ring A.
  • L is a bond.
  • Ring B is selected from the group consisting of: , 5 , 23
  • R 8 is C 1-5 alkyl which is optionally substituted with from 1-6 independently 5 selected R 12 .
  • R 8 is C 1-3 alkyl which is optionally substituted with from 1-2 independently selected R 12 .
  • R 8 is selected from the group consisting of: methyl, propyl, and isopropyl each of which may be optionally substituted with from 1-2 independently selected R 12 .
  • R 12 is selected from the group consisting of -C1-4 alkoxy; and -OH. 15 In some embodiments, R 12 is selected from the group consisting of methoxy and -OH.
  • Ring B is selected from the group consisting of: , , 24
  • the compound is a compound of Formula (VI): 25
  • Ring A is selected from the group consisting of: • , wherein * is the point of attachment to L, wherein X 1 and X 2 are each selected from the group consisting of CH and N and wherein a maximum of one of X 1 and X 2 may be N; or • , wherein Z is the point of attachment to L, Y is selected from the 10 group consisting of CH and N, and wherein Z is N;
  • Ring B is selected from the group consisting of: • heterocyclyl or heterocycloalkenyl including 5-9 ring atoms, wherein from 1-3 ring atoms are heteroatoms, each independently selected from the group consisting of N, N(H), N(R 1 ) and O, wherein one or more of the carbon atoms of the heterocyclyl or 15 heterocycloalkenyl is optionally substituted with from 1-4 substituents independently selected from the group consisting of oxo and R 8
  • the compound is selected from the group consisting of the compounds in Table 1, or a pharmaceutically acceptable salt thereof.
  • Pharmaceutical Compositions 5 the present disclosure provides a pharmaceutical composition that includes any one of the compounds described herein, or a pharmaceutically acceptable salt thereof (e.g., a therapeutically effective amount of the compound or salt), and a pharmaceutically acceptable excipient.
  • compositions provided herein can be administered by a variety of 10 routes including, but not limited to, oral (enteral) administration, parenteral (by injection) administration, rectal administration, transdermal administration, intradermal administration, intrathecal administration, subcutaneous (SC) administration, intravenous (IV) administration, intramuscular (IM) administration, and intranasal administration. Details of such formulations and processing thereof are set forth in Part 8 of Remington’s Pharmaceutical Sciences, 17th edition, 15 1985, Mack Publishing Company, Easton, Pennsylvania, which is incorporated herein by reference. Methods of Use In one aspect, this disclosure features methods of degrading VAV1 in a subject, which 20 include administering to the subject an effective amount of a compound described herein, or pharmaceutically acceptable salt thereof.
  • the compounds described herein can bind to a specific amino acid sequence of VAV1, thereby causing degradation of VAV1.
  • the compound mediates the interaction of a VAV1 protein with an E3 ligase, thereby increasing degradation of the VAV1 protein.
  • VAV1 is a regulator of a lymphocyte. 25
  • the compound interacts with the E3 ligase prior to the interaction of VAV1 with the E3 ligase.
  • the E3 ligase comprises cereblon. Degradation of VAV1 is mediated by the compound interacting with both the specific amino acid sequence of VAV1 and an E3 ligase.
  • this disclosure features methods of degrading VAV1, which include: (i) contacting a compound described herein or a 30 pharmaceutically acceptable salt thereof with an E3 ligase; and (ii) interacting the contacted E3 ligase with VAV1, thereby degrading VAV1.
  • the E3 ligase comprises cereblon.
  • this disclosure features methods of treating a variety of disorders which include administering the compounds and pharmaceutical compositions described herein. Such 27
  • PAT059977-WO-PCT disorders include, without limitation, autoimmune diseases (e.g., multiple sclerosis, rheumatoid arthritis, myasthenia gravis), transplantation setting disease (e.g., graft-versus-host disease), and cancers, tumours or other malignancies, (e.g. a T cell or B cell malignancy).
  • autoimmune diseases e.g., multiple sclerosis, rheumatoid arthritis, myasthenia gravis
  • transplantation setting disease e.g., graft-versus-host disease
  • cancers, tumours or other malignancies e.g. a T cell or B cell malignancy
  • this disclosure features methods of treating a disorder caused by or associated 5 with disregulation of lymphocyte development or activation in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound described herein or a pharmaceutically acceptable salt thereof.
  • the disorder is autoimmune disease (e.g., multiple sclerosis, psoriatic arthritis, rheumatoid arthritis, myasthenia gravis).
  • the disorder is transplantation setting disease (e.g., graft-versus- 10 host disease).
  • the disorder is a cancer or tumor.
  • the disorder is a malignancy (e.g., T cell or B cell malignancy).
  • the lymphocyte is T-cell. In some embodiments, the lymphocyte is B-cell.
  • this disclosure features methods of treating a disorder caused by or associated with dysregulation of T-cell receptor signaling in a subject in need thereof, comprising 15 administering to the subject a therapeutically effective amount of a compound described herein or a pharmaceutically acceptable salt thereof.
  • the T-cell receptor signaling are enhanced CD69 surface expression, IFN ⁇ or IL-2.
  • this disclosure features methods of treating a disorder caused by or associated with VAV1 polymorphisms in a subject in need thereof, comprising administering to the subject 20 a therapeutically effective amount of a compound described herein or a pharmaceutically acceptable salt thereof.
  • this disclosure features methods of treating a disorder caused by or associated with immunopathologies in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound described herein or a pharmaceutically acceptable 25 salt thereof.
  • the disorder is autoimmune disorder.
  • the disorder is a cancer, tumour or other malignancy, optionally wherein the disorder is a T cell or B cell malignancy.
  • the disorder is selected 5 from the group consisting of: leukemia, lymphoma, T-cell prolymphocytic leukemia, T-cell granular lymphocytic leukemia, aggressive NK cell leukemia, hairy-cell leukemia, nasal and nasal- type NK/T cell lymphoma, mycosis fungoides and Sezary syndrome, angioimmunoblastic T-cell lymphoma, peripheral T-cell lymphoma unspecified, adult T-cell leukemia/lymphoma (HTLV1+), anaplastic large cell lymphoma, primary cutaneous CD-30 positive T-cell lymphoproliferative 10 disorders, cutaneous T-cell lymphoma, subcutaneous panniculitis like T-cell lymphoma, intestinal T-cell lymphoma (+enteropathy), hepatosplenic gamma/delta T-cell lymphoma, and non-Hodgkin lymphomas (e.g., B-cell non-Hod
  • the disorder 20 is selected from the group consisting of Diabetes Type I or II, pernicious anemia, uveitis, psoriasis, alopecia areata, ulcerative colitis, Chron’s disease, atherosclerosis, myocarditis, pericarditis, pulmonary fibrosis, systemic sclerosis, morphea, Alzheimer’s disease, Acute Graft-vs. Host Disease or T-cell mediated kidney disease. In some embodiments, the disorder is T/B-cell mediated.
  • the 25 disorder is selected from the group consisting of multiple sclerosis, psoriatic arthritis, rheumatoid arthritis, myasthenia gravis, Sjogren’s syndrome, Grave’s disease, an allergic disorder (e.g., asthma, allergic contact dermatitis, rhinitis or contact dermatitis), an autoimmune liver disease (e.g., biliary sclerosis or sclerosing cholangitis), chronic inflammatory demyelinating polyradiculoneuropathy, macular degeneration, systemic lupus erythematosus, Hashimoto’s 30 thyroiditis, amyloidosis, inflammatory eye diseases, pemphigus, systemic lupus erythematosus, Chronic Graft vs.
  • an allergic disorder e.g., asthma, allergic contact dermatitis, rhinitis or contact dermatitis
  • an autoimmune liver disease e.g., biliary sclerosis or sclerosing
  • the disorder is selected from the group consisting of ulcerative colitis, psoriatic arthritis, rheumatoid arthritis, psoriasis, multiple sclerosis, myasthenia gravis, cutaneous lupus or axial spondylarthritis. 29
  • the disorder is selected from the group consisting of B-cell lymphoma, B-cell leukemia, T-cell lymphoma, T-cell leukemia or acute myeloid leukemia.
  • the disorder is a non-Hodgkin lymphoma.
  • the disorder is selected from mantle cell lymphoma (MCL), chronic lymphocytic leukemia (CLL) 5 and Diffuse large B cell lymphoma (DLBCL).
  • the disorder is chronic lymphocytic leukemia (CLL).
  • the disorder is multiple sclerosis (MS).
  • the disclosure relates to a method of treating patients exhibiting CD226 overexpression. 10 In some embodiments, the disclosure relates to a method of treating patients having a CD226 risk variant. In some embodiments, the disclosure relates to a method of treating patients having a CD226 polymorphism. In some embodiments, the disclosure relates to a method of treating patients having a 15 Gly307Ser (G307S) amino acid substitution in CD226 (rs763361T allele). In an aspect, the disclosure provides a compound or pharmaceutically acceptable salt for use in any of the above-recited methods of treatment. In some embodiments, the compound or pharmaceutically acceptable salt thereof is a compound or pharmaceutically acceptable salt 20 thereof as described herein.
  • PAT059977-WO-PCT mediates the coupling of the linker with an accessible amino residue on the binding moiety.
  • enzymatic conjugation include, but are not limited to, transpeptidation using sortase, transpeptidation using microbial transglutaminase, and N-glycan engineering. Chemical conjugation and enzymatic conjugation may also be used sequentially. For example, enzymatic 5 conjugation can also be used for installing unique reaction handles on Bm to be utilized in subsequent chemical conjugation.
  • M is a linker as defined in WO 2023/037268, which is incorporated by reference in its entirety.
  • M is selected from the group consisting of 10 wherein: q is from 2 to 10; Z 1 , Z 2 , Z 3 , Z 4 , and Z 5 are each independently absent or a naturally-occurring amino acid residue in the L- or D-configuration, provided that at least two of Z 1 , Z 2 , Z 3 , Z 4 , and Z 5 are amino acid 15 residues; is the point of attachment to the parent molecular (degrader) moiety; and is the point of attachment to the binding moiety.
  • an antibody is a protein generated by the immune system that is capable of recognizing and binding to a specific antigen.
  • a target antigen generally has numerous binding sites, also called epitopes, recognized by CDRs on multiple antibodies. Each antibody that specifically binds to a different epitope has a different structure. Thus, one antigen may have more 5 than one corresponding antibody.
  • the term "antibody” herein is used in the broadest sense and specifically covers monoclonal antibodies, single domain antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, so long as they exhibit the desired biological activity. Antibodies may be murine, human, humanized, chimeric, or derived from other species.
  • a monoclonal antibody (mAb) to an antigen-of-interest can be 10 prepared by using any technique known in the art which provides for the production of antibody molecules by continuous cell lines in culture. These include, but are not limited to, the hybridoma technique, the human B cell hybridoma technique, and the EBV-hybridoma technique. Such antibodies may be of any immunoglobulin class including IgG, IgM, IgE, IgA, and IgD and any subclass thereof.
  • the hybridoma producing the mAbs of use in this disclosure may be cultivated 15 in vitro or in vivo. The skilled person would understand how to provide an appropriate binding moiety for use in a conjugate depending on the intended therapeutic use.
  • an antibody, antibody fragment or an antibody-binding fragment used as a binding moiety must be capable of targeting a particular cell surface marker or receptor associated with the disorder to be treated.
  • the antibody trastuzumab can be employed if the desired target is HER2. 25
  • the binding moiety is capable of binding to an antigen selected from ⁇ 4 ⁇ 7, CD3, CD4, CD20, OX40, CD28, PD-1, ICOS, BCMA/TACl, CD52, CD30, CD19, CCR8, CD79b, CD22, CD4, CD7, CD38, CD33 or CD8 or combinations thereof.
  • the binding moiety is capable of binding to an antigen selected from CD19, CD20, CD33, CD4, CD8, PD-1, ICOS and CD38.
  • the binding moiety comprises an antibody selected from Vedolizumab, Etrolizumab, Teplizumab, Zanolimumab, Rituximab, Ublituximab, Ofatumumab, Ocrelizumab, Inebilizumab, Rocatinlimab, Nivolumab, Pembrolizumab, Alemtuzumab, Brentuximab vedotin, Tafasitamab, Loncastuximab, Mogamulizumab, Polatuzumab, Inotuzumab, Epratuzumab, 35 Isatuximab and Daratumumab. 34
  • the binding moiety is capable of binding to CD19 and is preferably Tafasitamab, Loncastuximab or Inebilizumab. In some embodiments, the binding moiety is capable of binding to CD20 and is preferably Rituximab, Ublituximab, Ofatumumab, Ocrelizumab 5 or Inebilizumab. In some embodiments, the binding moiety is capable of binding to CD33. In some embodiments, the binding moiety is capable of binding to CD4 and is preferably Zanolimumab. In some embodiments, the binding moiety is capable of binding to CD8.
  • the binding moiety is capable of binding to PD-1 and is preferably Nivolumab or Pembrolizumab. In some embodiments, the binding moiety is capable of binding to ICOS. In some embodiments, 10 the binding moiety is capable of binding to CD38 and is preferably Isatuximab or Daratumumab.
  • the method comprises administering the antibody-drug conjugate to the subject.
  • the binding moiety of the antibody-drug conjugate comprises an antibody listed in the table below and targets an antigen listed in the table below.
  • the disclosure provides a method of treating a disorder listed in the table below 10 comprising administering to a subject in need thereof an antibody-drug conjugate comprising an antibody listed in the table below.
  • the binding moiety of the antibody-drug conjugate comprises targets an antigen listed in the table below.
  • the disclosure provides a method of 5 treating a disorder listed in the table below comprising administering to a subject in need thereof an antibody-drug conjugate comprising a binding moiety which targets an antigen listed in the table below.
  • the disclosure provides a method of treating a disorder listed in the table below comprising administering to a subject in need thereof an antibody-drug conjugate comprising an antibody listed in the table below. 10 40
  • the antibody-drug conjugate may be administered as part of a pharmaceutical composition.
  • the pharmaceutical composition may include excipients such as those recited herein. 5
  • Non-Limiting Exemplary Compounds In some embodiments is a compound of Table 1 or a pharmaceutically acceptable salt thereof. Table 1: 41
  • PAT059977-WO-PCT A general synthetic strategy that may be used to prepare compounds of Formula II is depicted in General Scheme 1.
  • Compounds of Formula II may also be used as compounds of Formula I.
  • An aryl halide AA where Hal is any suitable halogen (e.g. Br or I), may be coupled with an aryl 5 boronate AB using any suitable metal catalyzed coupling conditions.
  • the specific groups X 1 , X 2 , X 3 , L 1 , R 2 , R 3 , R 4 , R 5 , R 6A , and Ring B are selected on the basis of the desired groups in the compound of Formula II.
  • DPPF 1,1′-Ferrocenediyl- bis(diphenylphosphine)
  • a solvent 10 mixture such as dioxane and water may be used.
  • compounds of Formula II may be prepared from reaction of an aryl boronate of formula AC and an aryl halide of formula AD using Suzuki cross-coupling conditions.
  • LG1 is any leaving group (for example, Br, I or triflate) which can be used in a metal catalyzed coupling reaction of AD to boronate AC.
  • Aryl boronate AC may be prepared from aryl halides AA using Bis(pinacolato)diboron and a catalyst such as Pd(dppf)Cl 2 .
  • a weak base such as potassium acetate in a solvent such as dioxane may be used.
  • the reaction may be performed at an elevated temperature, for example 85 degrees Celsius.
  • PAT059977-WO-PCT General Scheme 2 provides a synthetic procedure to prepare compounds of Formula IIIA. 5 Compounds of Formula IIIA may also be used as compounds of Formula I.
  • Intermediate AE can be coupled with a coupling partner such as AF to obtain intermediate AG, where Y and Z are nucleophiles suitable for a Buchwald-Hartwig coupling reaction, such as amines.
  • PG1 is any suitable protecting group that is labile to treatment with acid.
  • Intermediate AH may be obtained from AG upon treatment with an acid, such as HCl 2M in dioxane, at room 10 temperature.
  • a solvent such as dioxane may be used. 47
  • Intermediate AK may be obtained from AJ upon treatment with an acid, such as HCl 2M in dioxane, at room temperature.
  • a solvent such as DMF may be used.
  • Coupling partners AL are pyridones, where Hal is any halogen (e.g. Br or I) suitable for metal catalyzed coupling 20 conditions.
  • PAT059977-WO-PCT such as ruthenium (IV) oxide hydrate in the presence of sodium periodate.
  • Intermediate DE may be obtained from DC upon treatment with an acid, such as HCl 2M in dioxane, at room temperature.
  • Compounds of formula DG can be prepared from intermediates DE and DF via a coupling reaction with a copper(I) catalyst such as Cu(I)I in the presence of a base such as 5 potassium phosphate.
  • a solvent such as NMP may be used.
  • General Scheme 9 provides exemplary synthetic strategies for the preparation of aryl halides of 10 formula EB, which may be used as starting materials AD in General Scheme 1.
  • Hal is any suitable halogen (e.g.
  • General Scheme 10 provides exemplary synthetic strategies for the preparation of compounds of formula FB, which may be used as starting materials EA in General Scheme 9.
  • W 4 is a substituent containing a carbamate (for example, NH-Boc).
  • Compounds with the formula FB can 52
  • Step 2 Intermediate B-1 A mixture of Intermediate B-0 (1.00 g, 3.59 mmol, 1.00 eq.) and urea (668 mg, 11.1 mmol, 3.10 25 eq.) in acetic acid (10 mL) was stirred at 120°C for 12 h.
  • PAT059977-WO-PCT 2 190 mg, 434 ⁇ mol, 2.00 eq.
  • potassium phosphate 138 mg, 650 ⁇ mol, 3.00 eq.
  • 5 Pd(dppf)Cl2 16.0 mg, 21.9 ⁇ mol, 0.10 eq.
  • the mixture was cooled to room temperature and concentrated under reduced pressure to give a residue.
  • the residue was purified via Purification Method 2, then Purification Method 1 to afford Exemplary Compound 1 (36 mg, 75.0 ⁇ mol, 35% yield) as a white solid.
  • Example 24 B-2 (134 mg, 383 ⁇ mol, 2.00 eq.) and potassium phosphate (81.4 mg, 383 ⁇ mol, 2.00 eq.) were dissolved in DMF (2.0 mL) under nitrogen. Pd(dppf)Cl 2 (14.0 mg, 19.2 ⁇ mol, 0.10 eq.) was added, then the reaction was heated to 100°C and stirred until reaction 15 completion (2 h). The reaction mixture was quenched by adding water (8 mL) and the mixture was extracted with ethyl acetate (3 ⁇ 10 mL).
  • Step 6.4-(4-hydroxyphenyl)-6-(2-methoxyethyl)-4,6-diazaspiro-[2.4]heptan-5-one (210 mg, 703 ⁇ mol, 1.00 eq., hydrochloride) and triethylamine (284 mg, 2.80 mmol, 3.99 eq.) were dissolved in dichloromethane (2 mL) under nitrogen. Trifluoromethanesulfonyl chloride (350 mg, 2.08 mmol, 2.96 eq.) was added at 0°C. The reaction was stirred at 0°C for 2 h, then it was concentrated under 10 reduced pressure to give a residue.
  • Step 5 To a solution of 6-ethyl-4-(4-methoxyphenyl)-4,6-diazaspiro[2.4]heptan-5-one (160 mg, 649 ⁇ mol, 1.00 eq.) in dichloromethane (1.00 mL) was added boron tribromide (2.00 M, 1.62 mL, 5.00 eq.) at 0°C. The reaction was stirred at 0°C for 1 h, then it was quenched with water (20 mL) and extracted with ethyl acetate (3 ⁇ 15 mL).
  • Step 1.2-cyclobutylideneacetonitrile (2.00 g, 21.5 mmol, 1.00 eq.) was dissolved in a mixture of ammonia/methanol (7.00 M, 30.7 mL, 10.0 eq.). The reaction was stirred at 100°C for 12 h. The mixture was concentrated under reduced pressure to give a residue. The residue was purified via Purification Method 2 to afford 2-(1-aminocyclobutyl) acetonitrile (1.70 g, 15.4 mmol, 71% yield) as a yellow oil.
  • Intermediate A-10 was obtained from 2-(1-aminocyclobutyl) acetonitrile in analogy to Intermediate A-16 (Steps 1-6), using methyl iodide as the nucleophile in Step 4.
  • Intermediate A-7 Intermediate A-7 was synthesized in analogy to intermediate A-10, while omitting alkylation Step 4. 25 Intermediate A-25 78
  • Intermediate A-25 was synthesized in analogy to intermediate A-10, starting from 2-(oxetan-3- ylidene)acetonitrile and using 2,2-difluoroethyl trifluoromethanesulfonate as the nucleophile in Step 4.
  • Intermediate A-28 5 Intermediate A-28 was synthesized in analogy to intermediate A-25, using ethyl trifluoromethane-sulfonate as the nucleophile in Step 4.
  • Intermediate A-33 Intermediate A-33 was synthesized in analogy to intermediate A-25, using methyl iodide as the nucleophile in Step 4. 10
  • Intermediate A-22 Step 1 was synthesized in analogy to intermediate A-10, starting from 2-(oxetan-3- ylidene)acetonitrile and using 2,2-difluoroethyl trifluoromethanesulfonate as the nucleophile in Step 4.
  • Intermediate A-28 5 Intermediate A-28 was synthesized in analogy to intermediate
  • tert-butyl (1-(2-hydroxyethyl)cyclopropyl)carbamate (1.10 g, 5.47 mmol, 1.00 eq.)
  • isoindoline-1,3-dione (1.06 g, 7.18 mmol, 1.31 eq.)
  • triphenylphosphine (1.87 g, 7.13 mmol, 1.30 eq.) were dissolved in tetrahydrofuran (10 mL) under nitrogen, and the mixture was cooled 15 to 0°C.
  • N-(1-(2-aminoethyl)cyclopropyl)-4-bromo-2-nitroaniline 207 mg, 655 ⁇ mol, 35% yield
  • Step 5 N-(1-(2-aminoethyl)cyclopropyl)-4-bromo-2-nitroaniline (207 mg, 690 ⁇ mol, 1.00 eq.) was dissolved in tetrahydrofuran (2 mL) and 1,1'-carbonyldiimidazole (134 mg, 826 ⁇ mol, 1.20 eq.) was added.
  • tert-butyl 1-azaspiro[4.4]nonane-1-carboxylate (270 mg, 1.20 mmol, 1.00 eq.) was 20 dissolved in ethyl acetate (3.00 mL) and sodium periodate (1.28 g, 5.99 mmol, 5.00 eq.) was added, followed by a mixture of ruthenium (IV) oxide hydrate (54.3 mg, 359 ⁇ mol, 0.30 eq.) in water (3.00 mL). The reaction was stirred at 25°C for 2 h. The mixture was diluted with water (50 mL) and the aqueous layer was extracted with ethyl acetate (3 ⁇ 20 mL).
  • tert-butyl 2-oxo-1-azaspiro[4.4]nonane-1-carboxylate (280 mg, crude) as a colourless oil.
  • Step 3 A mixture of tert-butyl 2-oxo-1-azaspiro[4.4]nonane-1-carboxylate (280 mg, 1.17 mmol, 1.00 eq.) and trifluoroacetic acid (200 ⁇ L) in dichloromethane (2.00 mL) was stirred at 25°C for 1 h. The mixture was concentrated under reduced pressure to give residue. The residue was purified 83
  • Step 2 To a solution of 6-oxa-4-azaspiro[2.5]octan-5-one (170 mg, 1.34 mmol, 1.00 eq.) and 1- bromo-4-iodobenzene (491 mg, 1.74 mmol, 1.30 eq.) in N-methyl pyrrolidone (4.00 mL) were 10 added potassium phosphate (567 mg, 2.67 mmol, 2.00 eq.) and copper iodide (50.9 mg, 267 ⁇ mol, 0.20 eq.) under nitrogen. The reaction was stirred at 135°C for 12 h under nitrogen. The mixture was filtered, and the filtrate was concentrated under reduced pressure to give a residue.
  • Intermediate A-39 was synthesized in analogy to Intermediate A-34, using 2,2-difluoroethan-1- 10 amine as the amine partner in Step 1.
  • Intermediate A-60 Step 1. To a solution of 1-(2,2,2-trifluoroethyl)tetrahydropyrimidin-2(1H)-one (500 mg, 2.75 mmol, 1.00 eq.) in tetrahydrofuran (6 mL) was added sodium hydride 60% dispersion in mineral15 oil (329 mg, 8.24 mmol, 3.00 eq.) at 0°C.
  • tert-butyl (3-methyl-3-((4-nitrophenyl)amino)butyl)carbamate 800 mg, 2.47 mmol, 50% yield
  • Step 2 To a solution of tert-butyl (3-methyl-3-((4-nitrophenyl)amino)butyl)carbamate (800 mg, 2.47 mmol, 1.00 eq.) in tetrahydrofuran (1.00 mL) was added potassium tert-butoxide (832 mg, 7.42 mmol, 3.00 eq.). The reaction was stirred at 60°C for 0.5 h.
  • PAT059977-WO-PCT Step 1 To a solution of 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (2.53 g, 11.5 mmol, 1.50 eq.) in acetic acid (20 mL) were added potassium carbonate (2.12 g, 15.3 mmol, 2.00 eq.) and 3-(chloromethyl)-1-methyl-1H-pyrazole (1.00 g, 7.66 mmol, 1.00 eq.). The reaction was stirred at 65°C for 16 h. The mixture was filtered over Celite, and the filter cake was washed with acetonitrile 5 (100 mL). The filtrate was concentrated under reduce pressure to give a residue.
  • Example 68 Intermediate E (0.133 g, 0.387 mmol, 1.00 eq.) were dissolved in 5 dioxane (6 mL) and caesium carbonate (0.631 g, 1.94 mmol, 5.00 eq.) and Pd-PEPPSI-IHeptCl (37.6 mg, 38.7 ⁇ mol, 0.10 eq.) were added. The reaction was stirred at 100°C under nitrogen until reaction completion (12h). The mixture was diluted with water (20 mL), and the aqueous layer was extracted with ethyl acetate (3 ⁇ 20 mL).
  • PAT059977-WO-PCT a clonal Jurkat cell stably expressing LgBiT and possessing a homozygous GSPT1-G575N mutation.
  • Cells were plated at 10,000 cells per well using Multiflo (BioTek/Agilent) in 384-well white solid bottom plates (Corning, 3570BC) in 25 ul volume in RPMI 1640 media (Thermo Fischer, 22400105) containing 10% FBS (Corning, 35-075-CV), 1% Peniciliin/Streptomycin 5 (ThermoFisher Scientific, 15140-122), and 1% Endurazine (Nano-Glo Endurazine Live Cell Substrate (Promega, N2571)).
  • the luminescence response was then fitted in Genedata using a 4-parameter antagonist logistic fit 15 (hillslopeunconstrained, EC50 > 0, top/bottom unconstrained). The results of this study are presented in Table 8.
  • Examples B-J present biological activity data generated using compound 24 from Table 1. The same compound (“the VAV1 MGD”) was used in Examples B-J. 5
  • Example B VAV1 degradation in PBMCs after seven consecutive doses Mice were treated orally with seven consecutive doses of VAV1 MGD at 10, 1, and 0.1 mg/kg or vehicle.
  • Compound formulation was prepared fresh the day of administration in 10% Captisol in water. Serum and PBMCs were collected either 6 or 24 hours post-dosing for analysis 10 of pharmacokinetics and pharmacodynamics.
  • Serum samples were analyzed by LC-MS/MS to evaluate compound concentration in ng/mL and PBMCs were used to assess VAV1 protein levels and b-actin as loading control by western blotting.
  • PBMCs were rinsed with PBS then lysed using 100 ⁇ L RIPA lysis buffer (Pierce 89901) supplemented with 1% protease inhibitor cocktail [Roche 06493124001] and 1% phosphatase cocktail inhibitor [Sigma; P5726]) 15 respectively.
  • Samples were run on 4-12% precast gels (Thermo Fisher Scientific; WG1402BOX) at 10-15 ⁇ g/lane and transferred to a Transfer Stack (Thermo Fisher Scientific IB23001).
  • Example C VAV1 MGD induces degradation of VAV1 in lymphocytes and myeloid cells
  • Frozen human PBMCs (STEMCELL technologies; 70025.2; Lot: 2209401007 ) were thawed, washed with 1X PBS & resuspended in complete cell culture medium (RPMI + 10% FBS + 25mM HEPES + 1X Sodium Pyruvate (100X) + 1X MEM Non-Essential Amino Acids Solution 30 (100X) + 1% Penicillin-Streptomycin (10’000 U/mL) + 50 ⁇ M ⁇ -mercaptoethanol) .
  • Cells were plated at 1x10 6 cells/mL in U-bottom 96 well plate (Corning; 3799) and treated with VAV1 MGD 116
  • PAT059977-WO-PCT using a serial dilution ranging from 0.1 nM to 1000 nM and including a DMSO control. Plates were incubated for 24 hrs at 5% CO2, 37°C. Cells were then surface stained in FACS Buffer (PBS containing 10% FBS) with Live/Dead staining (Thermo Fisher Scientific; 34980) and Human TruStain FcX (BioLegend; 422302) for 20 minutes at 4oC. Cells were then washed with FACS 5 Buffer and surface antigens were stained in FACS Buffer (CD45, CD3, CD14, CD19) for 20 minutes at 4oC.
  • FACS Buffer PBS containing 10% FBS
  • Live/Dead staining Thermo Fisher Scientific; 34980
  • Human TruStain FcX BioLegend; 422302
  • Cells were then fixed with Cytofix (BD; 554655 ) and permeabilized with Phosflow Perm/Wash Buffer I (BD; 557885) and then and stained with anti-VAV1 antibody (CST; 2502) for 20 minutes at 4oC. Following staining with VAV1 antibody cells were washed with Perm buffer and incubated with Aleza Fluor 647-conjugated anti-rabbit IgG secondary 10 antibody (Biolegend; 406414) for 1 hour at 4oC. Cells were then washed with Perm/Wash Buffer and evaluated by flow cytometry. Cells were first gated on CD45 + , then respective cell marker (CD3 + , CD19 + , or CD14 + ).
  • Cells were plated 20 at optimized cell density in U-bottom 96 well plate (Costar; Z707899) and treated with VAV1 MGD using a serial dilution ranging from 0.01 nM to 1000 nM and including a DMSO control and incubated for 24 hrs at 5% CO2, 37°C. Cells were transferred from treatment plate to anti-CD3 coated plate (5 ⁇ g/mL anti-CD3 antibody, clone OKT3, Thermo Fisher; 16-0037-85) and co- stimulated with anti-CD28 (1 ⁇ g/mL, clone CD28.2, Thermo Fisher; 16-0289-85).
  • Cells were 25 incubated at 37°C with 5% CO2 and cells and/or supernatants harvested at various timepoints following anti-CD3/anti-CD28 co-stimulation for various readouts (24h: CD69; 48h: IL-2; 96h: proliferation).
  • CD69 surface evaluation cells were washed with PBS and stained for 20 minutes at 4°C with anti-CD69 allophycocyanin (APC) antibody (BioLegend ; 310910). Cells washed with PBS and evaluated by flow cytometry.
  • APC fluorophore signal intensity (CD69) 30 reported as % CD69 + cells normalized to anti-CD3/anti-CD28 co-stimulated DMSO control levels.
  • IL-2 was evaluated within cellular supernatants using an IL-2 ELISA kit (Abcam; ab270883) according to manufacturer protocol. Optical density (O.D.) values (IL-2) were obtained at 450 nM.
  • O.D. Optical density
  • IL-2 For proliferation assays, cells were labeled with Cell Trace Violet (Thermo Fisher Scientific; C34557) according to the manufacturer's instructions. Cells were analyzed as a percentage of 35 diluted Cell Trace Violet dye (relative to unstimulated cells) by flow cytometry. For all assays, 117
  • B cells were cultured in RPMI 1640 (Gibco; 22400089) supplemented with 10% fetal bovine serum (Gibco; #A31605-01; Lot 2408990P), 2mM L- glutamine (Gibco; 35050061), 100 IU/mL penicillin/streptomycin (Gibco; 15140122), 1 mM sodium pyruvate (Gibco, 11360070), 0.01 M HEPES (Gibco, 15630080), 1% non-essential amino acids (Gibco; 11140050), and 55 ⁇ M ⁇ -mercaptoethanol (Gibco; 21985023), and stimulated with 30 assay-dependent stimuli.
  • RPMI 1640 Gibco; 22400089
  • 10% fetal bovine serum Gibco; #A31605-01; Lot 2408990P
  • 2mM L- glutamine Gibco; 35050061
  • 100 IU/mL penicillin/streptomycin
  • B cells were stimulated with anti-IgM (1 ⁇ g/mL; Southern Biotech; 2022-14) and recombinant human IL-4 (10 ng/mL; R&D Systems; 305-IL-010) for 24 hrs.
  • IgG secretion B cells were stimulated with anti-IgM (1 ⁇ g/mL), BAFF (30 ng/mL; R&D Systems; #7537-BF-025), IL-21 (100 ng/mL; PeproTech; 200- 2), and soluble CD40L (50 ng/mL; R&D Systems; 245-C) for 5 days.
  • cells 35 were blocked with TruStain FcX (BioLegend; 422302), stained with mouse anti-human CD19- 118
  • Example H VAV1 MGD Attenuates FcR-Mediated Activity in Primary Human Monocytes 10 Monocytes were first isolated from human blood leukopaks. Each leukopak was diluted by adding 40 mL of the leukopak contents into 60mL of 1x PBS into a disposable Nalgene 150 mL bottle. The diluted blood mixture was transferred evenly among three Accupsin tubes ( ⁇ 30mL/tube). The mixture in Accuspin tubes was centrifuged at 720 x g for 20 minutes without brake at room temperature. Peripheral blood mononuclear cells) were collected from the interphase 15 with a sterile transfer pipette and transferred into new 50 mL tubes.
  • PBMCs were washed with 1X PBS and counted using Nexcelom cell counter. From the isolated PBMCs, monocytes were isolated using Pan Monocyte Isolation Kit (Miltenyi Biotec; 130-096-537) following the manufacturer’s instructions. Cells (1x106/mL) were treated for 24 hours in X-VIVO15 (Lonza; 02-053Q) supplemented with 10% fetal bovine serum with VAV1 MGD at the indicated final 20 concentrations or DMSO control and incubated at 37°C in a humidified incubator with 5% CO2 .
  • X-VIVO15 Longza; 02-053Q
  • VAV1 MGDs Degrade VAV1 in REC-1 Cell Line VAV1 degradation in REC-1 cells was measured by western blot quantification. REC-1 cells were seeded in RPMI-1640 (Gibco, CAT# A10491-01) + 10% FBS (Corning, CAT# 35- 016-CV, LOT# 16821001) at a density of 1 x 10 6 cells/mL and treated with a dose response of 35 VAV1 MGDs for 24 hours. Then, cells were harvested, washed with PBS and lysed using RIPA 119
  • PAT059977-WO-PCT buffer (Thermo, CAT# 89901) supplemented with protease inhibitor (Sigma, CAT# P8340) and phosphatase inhibitors (Sigma, CAT# P5726 + P0044). Protein concentration was determined with BCA Rapid Gold (Thermo, CAT# A53225) and subsequently normalized to 1.25 ⁇ g/ ⁇ L.
  • Samples were prepared with 1X NuPAGETM LDS Sample Buffer (Invitrogen, CAT# NP0007) + 5 5% DTT (Sigma, CAT# 43816) and denaturated at 95°C for 5 minutes.12 ⁇ L of each samples was loaded onto a 26-well Gel (Bio-Rad, CAT# 5678095) and run for ⁇ 45 minutes at 150 V. Then, proteins were transferred to a nitrocellulose membrane (Bio-Rad, CAT# 1704159) using the TurboTM Transfer System (Bio-Rad, CAT# 1704150) with 25 V for 7 minutes.
  • the membrane was blocked with EveryBlot Blocking Buffer (Bio-Rad, CAT# 12010020) followed 10 by overnight incubation with anti-VAV1 (CST, CAT# 2502) and anti- ⁇ -actin (CST, CAT# 3700S) primary antibodies. Then, the membrane was 3x washed with 1X TBS/T (Bio-Rad, CAT# BUF028) and incubated with anti-rabbit IgG HRP (Invitrogen, CAT# A16110) and anti- mouse IgG HRP (Invitrogen, CAT# 31432) secondary antibody followed by 3x wash with 1X TBS/T.
  • EveryBlot Blocking Buffer Bio-Rad, CAT# 12010020
  • anti-VAV1 CST, CAT# 2502
  • CST, CAT# 3700S anti- ⁇ -actin
  • Example J VAV1 MGDs Inhibit REC-1 Cell Line Proliferation 20
  • 3500 cells per well were plated in 75 ⁇ L medium of RPMI-1640 (Gibco; #A10491-01) supplemented with 10% FBS (Corning; #35-016-CV) and 100 U/mL penicillin/streptomycin (Gibco; #15140122) in 384- well plates (Corning; #3570) and settled overnight in a 37oC incubator with 5% CO 2 .
  • the cells were treated the following day with increasing concentrations of VAV1 MGDs.
  • DMSO was used 25 as a control. Each treatment was performed in triplicate.
  • CellTiter Glow (Promega; #G7573) readouts were performed as per the manufacturer’s instructions on Day 0 (pre-treatment) and Day 5 by adding 15 ⁇ L CTG reagent (Promega; #G7573) per well, incubating at room temperature for 5 minutes, followed by luminescence measurement using a PHERAstar FSX (BMG Labtech).
  • Example K Oral dosing of VAV1 MGDs inhibit EAE disease progression in a dose- dependent manner 5
  • EAE experimental autoimmune encephalomyelitis
  • J C57BL/6 Beijing Vital River Laboratory Animal Co; #213 mice were injected subcutaneously with an emulsified mixture consisting of 100 ⁇ g of the synthetic peptide derived from myelin oligodendrocyte glycoprotein (MOG35-55; GL Biochem Ltd; #51716)) and 200 ⁇ g M.
  • tuberculosis (Difco; #231141) were mixed with incomplete Freund’s adjuvant (Sigma-Alrich; 10 #F5506) using a high-speed homogenizer (IKA T10 basic).
  • Mice were additionally injected intraperitoneally with 200 ng pertussis toxin (List Biological Laboratories; #180235AIA) at 0 and 48 hours post immunization. This immunization induces the activation and expansion of peripheral myelin-specific encephalitogenic T cells and their migration into the CNS. Once in the CNS, the activated T-cells initiate an inflammatory cascade, which ultimately leads to myelin 15 destruction and symptoms of paralysis.
  • vehicle 10% captisol in water
  • Compound 1 10 mg/kg or 20 Compound 24
  • VAV1 MGDs resuspended in 10% captisol in water
  • VAV1 MGDs were formulated fresh the day of administration by dissolving the VAV1 MGDs in 10% captisol in water then vortexed until a clear solution or a uniform suspension was obtained.
  • mice were euthanized then 25 peripheral blood mononuclear cells (PBMC) were enriched from cardiac puncture samples and brain tissue was excised then snap frozen for subsequent assessment of VAV1 levels by western blot.
  • PBMC peripheral blood mononuclear cells
  • tissue samples were defrosted, 30-100 mg of tissue was placed in a 2 mL microcentrifuge tube, and 400 ⁇ L RIPA buffer (Sigma; #R0278) containing 1% 30 protease inhibitor cocktail (Roche; #04693124001) and 1% phosphatase inhibitor cocktail 2 (Sigma; #P5726) was added to each tube.
  • RIPA buffer Sigma; #R0278
  • 1% 30 protease inhibitor cocktail (Roche; #04693124001)
  • 1% phosphatase inhibitor cocktail 2 Sigma; #P5726
  • Tissues were ground using a Tissuelyser (Shanghai Jingzin; #JXSTPRP-CL) at 50 hZ for 5 mins. Samples were then incubated on ice for 30 mins.
  • samples were centrifuged at 13, 523 x g for 10 mins at 4oC, then transferred into new, pre-chilled microcentrifuge tubes.
  • frozen samples were 35 defrosted, placed in a 2 mL microcentrifuge tube, and 400 ⁇ L RIPA buffer (Sigma; #R0278) 121
  • PAT059977-WO-PCT containing 1% protease inhibitor cocktail (Roche; #04693124001) and 1% phosphatase inhibitor cocktail 2 (Sigma; #P5726) was added to each tube, then sonicated. Samples were then incubated on ice for 30 mins. After incubation, samples were centrifuged at 13, 523 x g for 10 mins at 4oC, then transferred into new, pre-chilled microcentrifuge tubes.
  • Protein concentrations were determined using a BCA assay kit (Thermo Fisher; #23225)) and samples were diluted to a final concentration of 2 ⁇ g/ ⁇ L in RIPA buffer containing 4X LDS sample buffer (Invitrogen; #NP0007) and 10X sample reducing agent (Invitrogen; #NP0009). Once diluted, samples were boiled at 100oC for 10 mins. Denatured samples were then stored at - 80oC. For western blotting, protein samples were defrosted and 15 ⁇ L of each sample was loaded 10 in 4-12% Bis-Tris gels (Invitrogen; #WG1402BOX).
  • Electrophoresis was then run in MES running buffer (Invitrogen; #NP0002) at 80 V for 30 mins then 120 V for 90 mins. Proteins were transferred to a nitrocellulose membrane with an iBlot 2 Gel Transfer Device (Invitrogen) using P3 for 7 mins. After transfer, membranes were washed (10 mL 1X TBST, 5 mins, 3 times), blocked (TBS Blocking Buffer (LI-COR;#927-60001), 1 h with agitation, room temperature), 15 and washed again (1X TBST (Bio-Serve; #BS-P-15), 10 mins, 3 times).
  • VAV1 and ⁇ -actin were detected by incubation with anti-VAV1 (CST; 2502) and ⁇ -actin (CST; #4967) antibodies (1:1000 or 1:2000 respectively in TBS Blocking Buffer containing 0.1% Tween-20 (Sigma; #P2287) at 4oC overnight with gentle agitation).
  • Membranes were then washed (10 mL 1X TBST, 10 mins, 3 times) and then incubated with secondary detection goat anti-mouse IgG-20 IRDye 860RD (LI-COR; #926-68070) and goat anti-rabbit IgG-IRDye 800CW (LI-COR; #926- 32211) antibodies (both 1:10000 in TBD Blocking Buffer containing 0.1% Tween-20 for 1h at room temperature with gentle agitation, protected from light). Membranes were then washed (10 mL 1X TBST, 5 mins, 5 times) and protein levels were detected by fluorescence signal using an Odessey CLx Imaging System (LI-COR).
  • LI-COR Odessey CLx Imaging System
  • VAV1 protein levels were quantified by fluorescence 25 intensity relative to ⁇ -actin and normalized to the vehicle treatment group. The results of this study are shown in FIGS.10A, 10B, and 10C. These data show that both Compound 1 and Compound 24 inhibit disease progression in a mouse model of EAE, with dose-dependent activity observed with Compound 24 (FIG.10A), consistent with degradation observed in PBMCs. Both Compound 1 and Compound 24 degrade 30 VAV1 in peripheral PBMCs, with dose-dependent activity observed with Compound 24 (FIG. 10B), but not within brain tissue (FIG.10C) due peripheral-restriction of these compounds. Therefore, degradation of VAV1 in peripheral PBMCs is sufficient to inhibit disease progression of a T cell-mediated EAE model. 122

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Abstract

La présente divulgation concerne des entités chimiques (par exemple, un composé ou un sel pharmaceutiquement acceptable de celui-ci) qui dégradent la protéine VAV 1 (VAV1) proto-oncogène. Les entités chimiques sont utiles pour traiter des sujets présentant un trouble ou une maladie qui peuvent être traités par réduction du taux de VAV1, tels que le cancer, les troubles inflammatoires ou auto-immuns.
PCT/IB2025/056919 2024-07-10 2025-07-08 Dérivés de dihydrouracile utiles pour la dégradation ciblée de vav1 Pending WO2026013576A1 (fr)

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Citations (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2021058017A1 (fr) * 2019-09-29 2021-04-01 Beigene, Ltd. Dégradation du récepteur des androgènes (ar) par conjugaison d'antagonistes ar avec un ligand de ligase e3 et procédés d'utilisation
WO2021198966A1 (fr) 2020-03-31 2021-10-07 Orum Therapeutics, Inc. Conjugués
US20220363671A1 (en) * 2019-09-16 2022-11-17 Novartis Ag Glue degraders and methods of use thereof
WO2023017446A1 (fr) * 2021-08-10 2023-02-16 Uppthera, Inc. Nouveau composé induisant la dégradation de plk1
WO2023037268A1 (fr) 2021-09-08 2023-03-16 Orum Therapeutics, Inc. Lieurs destinés à être utilisés dans des conjugués anticorps-médicament
EP4342892A1 (fr) * 2021-07-01 2024-03-27 Hangzhou Healzen Therapeutics Co., Ltd. Tyrosine kinase de bruton et agent de dégradation mutant, composition et utilisation associées
WO2024083255A1 (fr) * 2022-10-21 2024-04-25 上海领泰生物医药科技有限公司 Dérivé d'aminoquinazoline substitué par benzyle ou thiénylméthylène et son utilisation en tant qu'agent de dégradation de sos1
WO2024098692A1 (fr) * 2022-11-08 2024-05-16 杭州格博生物医药有限公司 Agent de dégradation de protéine kinase wee1 et son utilisation
EP4393922A1 (fr) * 2021-08-23 2024-07-03 Shanghai Leadingtac Pharmaceutical Co. Ltd. Agent de dégradation d'irak4, son procédé de préparation et son utilisation
WO2024151547A1 (fr) * 2023-01-09 2024-07-18 Monte Rosa Therapeutics, Inc. Dégradation ciblée de vav1

Patent Citations (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20220363671A1 (en) * 2019-09-16 2022-11-17 Novartis Ag Glue degraders and methods of use thereof
WO2021058017A1 (fr) * 2019-09-29 2021-04-01 Beigene, Ltd. Dégradation du récepteur des androgènes (ar) par conjugaison d'antagonistes ar avec un ligand de ligase e3 et procédés d'utilisation
WO2021198966A1 (fr) 2020-03-31 2021-10-07 Orum Therapeutics, Inc. Conjugués
EP4342892A1 (fr) * 2021-07-01 2024-03-27 Hangzhou Healzen Therapeutics Co., Ltd. Tyrosine kinase de bruton et agent de dégradation mutant, composition et utilisation associées
WO2023017446A1 (fr) * 2021-08-10 2023-02-16 Uppthera, Inc. Nouveau composé induisant la dégradation de plk1
EP4393922A1 (fr) * 2021-08-23 2024-07-03 Shanghai Leadingtac Pharmaceutical Co. Ltd. Agent de dégradation d'irak4, son procédé de préparation et son utilisation
WO2023037268A1 (fr) 2021-09-08 2023-03-16 Orum Therapeutics, Inc. Lieurs destinés à être utilisés dans des conjugués anticorps-médicament
WO2024083255A1 (fr) * 2022-10-21 2024-04-25 上海领泰生物医药科技有限公司 Dérivé d'aminoquinazoline substitué par benzyle ou thiénylméthylène et son utilisation en tant qu'agent de dégradation de sos1
WO2024098692A1 (fr) * 2022-11-08 2024-05-16 杭州格博生物医药有限公司 Agent de dégradation de protéine kinase wee1 et son utilisation
WO2024151547A1 (fr) * 2023-01-09 2024-07-18 Monte Rosa Therapeutics, Inc. Dégradation ciblée de vav1

Non-Patent Citations (8)

* Cited by examiner, † Cited by third party
Title
"Remington's Pharmaceutical Sciences", 1985, MACK PUBLISHING COMPANY
BERGE ET AL., J. PHARMACEUTICAL SCIENCES, vol. 66, 1977, pages 1 - 19
HAUBERT ET AL., TRANSPLANTATION IMMUNOLOGY, vol. 26, 2012, pages 212
KOM ET AL., JOURNAL OF NEUROIMMUNOLOGY, vol. 139, 2003, pages 17
LEMAITRE THOMAS ET AL: "Molecular glue degraders: exciting opportunities for novel drug discovery", EXPERT OPINION ON DRUG DISCOVERY, vol. 19, no. 4, 24 January 2024 (2024-01-24), London, GB, pages 433 - 449, XP093308842, ISSN: 1746-0441, Retrieved from the Internet <URL:https://www.tandfonline.com/doi/pdf/10.1080/17460441.2024.2306845> DOI: 10.1080/17460441.2024.2306845 *
MARC GERSPACHER: "discovery of selective low molecular weight vav1 guanine nucleotide exchange factor inhibitors", ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY,MEETING 2017, 254, ABSTRACT MEDI306, 20 August 2017 (2017-08-20), pages 1 - 1, XP093156446, Retrieved from the Internet <URL:https://www.morressier.com/o/event/5fc63e1403137aa5257641f7/article/5fc63eeb9e0a135cbed0d9b0> *
NATURE REVIEWS DRUG DISCOVERY VOLUME, vol. 22, 2023, pages 641 - 661
SCHMIDT ET AL., SCIENCE, vol. 375, 2022, pages 6580

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