L7019.019PV1
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`TLR AGONIST IMMUNOCONJUGATES WITH CYSTEINE-MUTANT ANTIBODIES,
`AND USES THEREOF
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`FIELD OF THE INVENTION
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`The invention relates generally to an immunoconjugate comprising a cysteine-mutant
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`antibody conjugated to one or moretoll-like receptor (TLR) agonist moieties.
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`BACKGROUNDOF THE INVENTION
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`New compositions and methods for the delivery of antibodies and immuneadjuvantsare
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`needed in order to reach inaccessible tumors and/or to expand treatment options for cancer
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`patients and other subjects. The invention provides such compositions and methods.
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`SUMMARYOF THE INVENTION
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`The invention is generally directed to an immunoconjugate comprising a cysteine-mutant
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`antibody covalently attached to one or more TLR agonist moieties by a linker.
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`Another aspect of the invention is a method of preparing an immunoconjugate by
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`conjugation of one or more TLR agonist-linker compounds with a cysteine-mutant antibody.
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`Anotheraspect of the invention is a pharmaceutical composition comprising a
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`therapeutically effective amount of an immunoconjugate comprising a cysteine-mutant antibody
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`covalently attached to one or more TLR agonist moieties by a linker, and one or more
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`pharmaceutically acceptable diluent, vehicle, carrier or excipient.
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`Another aspect of the invention is a method for treating cancer comprising administering
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`a therapeutically effective amount of an immunoconjugate comprising a cysteine-mutant
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`antibody covalently attached to one or more TLR agonist moieties by a linker.
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`Another aspect of the invention is a use an immunoconjugate comprising a cysteine-
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`mutant antibody covalently attached to one or more TLR agonist moieties by a linkerin the
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`treatment of an illness, in particular cancer.
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`BRIEF DESCRIPTION OF THE DRAWINGS
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`Figure 1 shows a graph demonstrating IL-12p70 secretion following activation of
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`enriched human cDCs (conventional dendritic cells) freshly isolated from human blood and co-
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`cultured with HCC 1954 tumorcells with immunoconjugates Lys IC-1 (Table 11), IC-2, IC-3,
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`IC-4, IC-8, IC-10, IC-13, IC-16, IC-17 and IC-18 (Table 10) and unconjugated antibody,
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`L7019.019PV1
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`trastuzumab. Logarithmic production of IL-12p70 is plotted at increasing concentrations
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`immunoconjugates and trastuzumab
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`Figure 2 shows a graph demonstrating IL-12p70 secretion following activation of
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`enriched human cDCs (conventional dendritic cells) freshly isolated from human blood and co-
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`cultured with HCC 1954 tumorcells with immunoconjugates IC-1, IC-12, IC-6, IC-11, IC-5, IC-
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`9, IC-7, IC-14, and IC-15 (Table 10), Lys IC-1 (Table 11), and unconjugated antibody,
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`trastuzumab. Logarithmic production of IL-12p70 is plotted at increasing concentrations of
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`immunoconjugates and trastuzumab.
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`DETAILED DESCRIPTION OF THE INVENTION
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`Reference will now be madein detail to certain embodiments of the invention, examples
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`of whichareillustrated in the accompanying structures and formulas. While the invention will
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`be described in conjunction with the enumerated embodiments, it will be understood that they
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`are not intendedto limit the invention to those embodiments. On the contrary, the invention is
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`intended to coverall alternatives, modifications, and equivalents, which may be included within
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`the scope of the invention as defined by the claims.
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`Oneskilled in the art will recognize many methods and materials similar or equivalent to
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`those described herein, which could be used in the practice of the present invention. The
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`invention is in no way limited to the methods and materials described.
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`DEFINITIONS
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`The terms “Toll-like receptor” and “TLR”refer to any memberof a family of highly-
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`conserved mammalian proteins which recognizes pathogen-associated molecular patterns and
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`acts as key signaling elements in innate immunity. TLR polypeptides share a characteristic
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`structure that includes an extracellular domain that has leucine-rich repeats, a transmembrane
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`domain, and an intracellular domain that is involved in TLR signaling.
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`The terms “Toll-like receptor 7” and “TLR7”refer to nucleic acids or polypeptides
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`sharing at least about 70%, about 80%, about 90%, about 95%, about 96%, about 97%, about
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`98%, about 99%, or more sequenceidentity to a publicly-available TLR7 sequence, e.g.,
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`GenBankaccession number AAZ99026 for human TLR7 polypeptide, or GenBank accession
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`number AAK62676 for murine TLR7 polypeptide.
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`The terms “Toll-like receptor 8” and “TLR8”refer to nucleic acids or polypeptides
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`sharing at least about 70%, about 80%, about 90%, about 95%, about 96%, about 97%, about
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`98%, about 99%, or more sequenceidentity to a publicly-available TLR7 sequence, e.g.,
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`GenBankaccession number AAZ95441 for human TLR8 polypeptide, or GenBank accession
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`number AAK62677 for murine TLR8 polypeptide.
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`L7019.019PV1
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`A “TLRagonist” is a compoundthat binds, directly or indirectly, to a TLR (e.g., TLR7
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`and/or TLR8) to induce TLR signaling. Any detectable difference in TLR signaling can indicate
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`that an agonist stimulates or activates a TLR. Signaling differences can be manifested, for
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`example, as changes in the expression of target genes, in the phosphorylation of signal
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`transduction components, in the intracellular localization of downstream elements such as
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`nuclear factor-«B (NF-kB), in the association of certain components (such as IL-1 receptor
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`associated kinase (IRAK)) with other proteins or intracellular structures, or in the biochemical
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`activity of components such as kinases (such as mitogen-activated protein kinase (MAPK)).
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`“Antibody” refers to a polypeptide comprising an antigen binding region (including the
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`complementarity determining region (CDRs)) from an immunoglobulin gene or fragments
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`thereof. The term “antibody” specifically encompasses monoclonal antibodies (including full
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`length monoclonal antibodies), polyclonal antibodies, multispecific antibodies (e.g., bispecific
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`antibodies), and antibody fragments that exhibit the desired biological activity. An exemplary
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`immunoglobulin (antibody) structural unit comprises a tetramer. Each tetramer is composed of
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`two identical pairs of polypeptide chains, each pair having one “light” (about 25 kDa) and one
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`“heavy” chain (about 50-70 kDa) connected by disulfide bonds. Each chain is composed of
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`structural domains, which are referred to as immunoglobulin domains. These domainsare
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`classified into different categories by size and function, e.g., variable domains or regions on the
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`light and heavy chains (V~ and Vu, respectively) and constant domainsor regions on thelight
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`and heavy chains (Cx and Cu, respectively). The N-terminus of each chain defines a variable
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`region of about 100 to 110 or more aminoacids, referred to as the paratope, primarily
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`responsible for antigen recognition, i.e., the antigen binding domain. Light chains are classified
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`as either kappa or lambda. Heavy chainsare classified as gamma, mu, alpha, delta, or epsilon,
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`which in turn define the immunoglobulin classes, IgG, IgM, IgA, IgD and IgE,respectively.
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`IgG antibodies are large molecules of about 150 kDa composedof four peptide chains. IgG
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`antibodies contain two identical class y heavy chains of about 50 kDa and twoidentical light
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`chains of about 25 kDa, thus a tetrameric quaternary structure. The two heavy chains are linked
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`to each other and to a light chain each by disulfide bonds. The resulting tetramer has two
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`identical halves, which together form the Y-like shape. Each end ofthe fork contains an
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`identical antigen binding domain. There are four IgG subclasses (IgG1, IgG2, IgG3, and IgG4)
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`in humans, named in order of their abundance in serum (i.e., IgG1 is the most abundant).
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`Typically, the antigen binding domain of an antibody will be mostcritical in specificity and
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`affinity of binding to cancercells.
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`“Bispecific” antibodies (bsAbs) are antibodies that bind two distinct epitopes to cancer
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`(Suurs F.V. et al (2019) Pharmacology & Therapeutics 201: 103-119). Bispecific antibodies
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`L7019.019PV1
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`may engage immunecells to destroy tumorcells, deliver payloads to tumors, and/or block tumor
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`signaling pathways. An antibody that targets a particular antigen includes a bispecific or
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`multispecific antibody with at least one antigen binding region that targets the particular antigen.
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`In some embodiments, the targeted monoclonal antibody is a bispecific antibody with at least
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`one antigen binding region that targets tumorcells. Such antigens include but are not limited to:
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`mesothelin, prostate specific membrane antigen (PSMA), HER2, TROP2, CEA, EGFR,
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`5T4,Nectin4, CD19, CD20, CD22, CD30, CD70, B7H3, B7H4(also knownas O8E), protein
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`tyrosine kinase 7 (PTK7), glypican-3, RG1, fucosyl-GMI, CTLA-4, and CD44 (WO
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`2017/196598).
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`“Antibody construct” refers to an antibody or a fusion protein comprising (i) an antigen
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`binding domain and(11) an Fe domain.
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`The term “immunoconjugate” refers to an antibody construct that is covalently bonded to
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`an adjuvant moiety via a linker. Immunoconjugates allowtargeted delivery of an active adjuvant
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`moiety while the target antigen is bound.
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`“Adjuvant” refers to a substance capable of eliciting an immuneresponse in a subject
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`exposed to the adjuvant. The phrase “adjuvant moiety” refers to an adjuvant that is covalently
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`bonded to an antibody construct, e.g., through a linker, as described herein. The adjuvant
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`moiety can elicit the immune response while bonded to the antibody construct or after cleavage
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`(e.g., enzymatic cleavage) from the antibody construct following administration of an
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`immunoconjugate to the subject.
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`In some embodiments, the antibody construct is an antigen-binding antibody “fragment,”
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`which comprises at least an antigen-binding region of an antibody, alone or with other
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`components that together constitute the antibody construct. Many different types of antibody
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`“fragments” are known in theart, including, for instance, (i) a Fab fragment, whichis a
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`monovalent fragment consisting of the VL, Va, CL, and CHi domains,(ii) a F(ab’ )2 fragment,
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`whichis a bivalent fragment comprising two Fab fragmentslinked by a disulfide bridge at the
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`hinge region, (iii) a Fv fragment consisting of the VL and Vy domainsof a single arm of an
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`antibody, (iv) a Fab’ fragment, which results from breaking the disulfide bridge of an F(ab’)
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`fragment using mild reducing conditions, (v) a disulfide-stabilized Fv fragment (dsFv), and (vi)
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`a single chain Fv (scFv), which is a monovalent molecule consisting of the two domainsof the
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`Fv fragment(i.e., Vit and Vu) joined by a synthetic linker which enables the two domainsto be
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`synthesized as a single polypeptide chain.
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`The antibody or antibody fragment can be part of a larger construct, for example, a
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`conjugate or fusion construct of the antibody fragment to additional regions. For instance, in
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`some embodiments, the antibody fragment can be fused to an Fc region as described herein.
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`In
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`L7019.019PV1
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`other embodiments, the antibody fragment (e.g., a Fab or scFv) can be part of a chimeric antigen
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`receptor or chimeric T-cell receptor, for instance, by fusing to a transmembrane domain
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`(optionally with an intervening linker or “stalk” (e.g., hinge region)) and optional intercellular
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`signaling domain. For instance, the antibody fragment can be fused to the gammaand/ordelta
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`chainsofa t-cell receptor, so as to provide a T-cell receptor like construct that binds PD-L1. In
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`yet another embodiment, the antibody fragment is part of a bispecific T-cell engager (BiTEs)
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`comprising a CD1 or CD3 binding domain and linker.
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`“Cysteine-mutant antibody” is an antibody in which one or more aminoacid residues of
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`an antibody are substituted with cysteine residues. A cysteine-mutant antibody may be prepared
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`from the parent antibody by antibody engineering methods (Junutula, et al., (2008b) Nature
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`Biotech., 26(8):925-932; Dornanet al. (2009) Blood 114(13):2721-2729; US 7521541; US
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`7723485; US 2012/0121615; WO 2009/052249). Cysteine residues provide for site-specific
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`conjugation of a adjuvant such as a TLR agonist to the antibody through the reactive cysteine
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`thiol groups at the engineered cysteine sites but do not perturb immunoglobulin folding and
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`assembly or alter antigen binding and effector functions. Cysteine-mutant antibodies can be
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`conjugated to the TLR agonist-linker compound with uniform stoichiometry of the
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`immunoconjugate (e.g., up to two TLR agonist moieties per antibody in an antibody that has a
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`single engineered, mutant cysteine site). The TLR agonist-linker compound has a reactive
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`electrophilic group to react specifically with the free cysteine thiol groups of the cysteine-mutant
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`antibody.
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`“Epitope” means any antigenic determinantor epitopic determinant of an antigen to
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`which an antigen binding domain binds(1.e., at the paratope of the antigen binding domain).
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`Antigenic determinants usually consist of chemically active surface groupings of molecules,
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`such as amino acids or sugar side chains, and usually have specific three dimensional structural
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`characteristics, as well as specific charge characteristics.
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`The terms “Fc receptor” or “FcR” refer to a receptor that binds to the Fc region of an
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`antibody. There are three main classes of Fc receptors: (1) FcyR which bind to IgG, (2) FcaR
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`which binds to IgA, and (3) FceR which binds to IgE. The FcyR family includes several
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`members,such as Feyl (CD64), FeyRIA (CD32A), FcyRIIB (CD32B), FcyRIIA (CD16A), and
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`FcyRIIB (CD16B). The Feyreceptors differ in their affinity for IgG and also havedifferent
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`affinities for the IgG subclasses (e.g., IgG1, IgG2, IgG3, and IgG4).
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`Nucleic acid or amino acid sequence “identity,” as referenced herein, can be determined
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`by comparing a nucleic acid or amino acid sequenceofinterest to a reference nucleic acid or
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`amino acid sequence. The percent identity is the numberof nucleotides or amino acid residues
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`that are the same(i.e., that are identical) as between the optimally aligned sequenceof interest
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`L7019.019PV1
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`and the reference sequence divided by the length of the longest sequence(i.e., the length of
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`either the sequence of interest or the reference sequence, whichever is longer). Alignment of
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`sequences and calculation of percent identity can be performed using available software
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`programs. Examples of such programs include CLUSTAL-W,T-Coffee, and ALIGN (for
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`alignment of nucleic acid and amino acid sequences), BLAST programs (e.g., BLAST 2.1,
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`BL2SEQ, BLASTp, BLASTn,and the like) and FASTA programs (e.g., FASTA3x, FASTM,
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`and SSEARCH)(for sequence alignment and sequence similarity searches). Sequence
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`alignment algorithms also are disclosed in, for example, Altschul et al., . Molecular Biol.,
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`215(3): 403-410 (1990), Beigert et al., Proc. Natl. Acad. Sci. USA, 106(10): 3770-3775 (2009),
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`Durbinet al., eds., Biological Sequence Analysis: Probalistic Models ofProteins and Nucleic
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`Acids, Cambridge University Press, Cambridge, UK (2009), Soding, Bioinformatics, 21(7): 951-
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`960 (2005), Altschul et al., Nucleic Acids Res., 25(17): 3389-3402 (1997), and Gusfield,
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`Algorithms on Strings, Trees and Sequences, Cambridge University Press, Cambridge UK
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`(1997)). Percent (%) identity of sequences can be also calculated, for example, as 100 x
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`[(identical positions)/min(TGa, TGp)], where TGa and TGgp are the sum of the number of
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`residues and internal gap positions in peptide sequences A and B in the alignment that
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`minimizes TGs and TGpg. See, e.g., Russell et al., J. Mol Biol., 244: 332-350 (1994).
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`The “antibody construct” or “binding agent” comprises Ig heavy and light chain variable
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`region polypeptides that together form the antigen binding site. Each of the heavy and light
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`chain variable regions are polypeptides comprising three complementarity determining regions
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`(CDR1, CDR2, and CDR3) connected by framework regions. The antibody construct can be
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`any of a variety of types of binding agents knownin the art that comprise Ig heavy and light
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`chains. For instance, the binding agent can be an antibody, an antigen-binding antibody
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`“fragment,” or a T-cell receptor.
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`“Biosimilar”’ refers to an approved antibody construct that has active properties similar
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`to, for example, a PD-L1-targeting antibody construct previously approved such as atezolizumab
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`(TECENTRIQ™, Genentech, Inc.), durvalumab (IMFINZI™, AstraZeneca), and avelumab
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`(BAVENCIO™, EMDSerono,Pfizer); a HER2-targeting antibody construct previously
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`approved such as trastuzumab (HERCEPTIN™, Genentech, Inc.), and pertuzumab
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`(PERJETA™, Genentech, Inc.); or a CEA-targeting antibody such as labetuzumab (CEA-
`CIDE™, MN-14, hMN14, Immunomedics) CAS Reg. No. 219649-07-7).
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`“Biobetter”’ refers to an approved antibody construct that is an improvementofa
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`previously approved antibody construct, such as atezolizumab, durvalumab, avelumab,
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`trastuzumab, pertuzumab, and labetuzumab. The biobetter can have one or more modifications
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`L7019.019PV1
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`(e.g., an altered glycan profile, or a unique epitope) over the previously approved antibody
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`construct.
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`“Amino acid” refers to any monomeric unit that can be incorporated into a peptide,
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`polypeptide, or protein. Aminoacidsinclude naturally-occurring a-amino acids and their
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`stereoisomers, as well as unnatural (non-naturally occurring) amino acids andtheir
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`stereoisomers. “Stereoisomers” of a given amino acid refer to isomers having the same
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`molecular formula and intramolecular bonds but different three-dimensional arrangements of
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`bonds and atoms (e.g., an L-amino acid and the corresponding D-amino acid). The amino acids
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`can be glycosylated (e.g., N-linked glycans, O-linked glycans, phosphoglycans, C-linked
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`glycans,or glypication) or deglycosylated. Amino acids may be referred to herein by either the
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`commonly knownthree letter symbols or by the one-letter symbols recommendedby the
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`IUPAC-IUB Biochemical Nomenclature Commission.
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`Naturally-occurring amino acids are those encoded bythe genetic code, as well as those
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`amino acids that are later modified, e.g., hydroxyproline, y-carboxyglutamate, and
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`O-phosphoserine. Naturally-occurring a-amino acidsinclude, without limitation, alanine (Ala),
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`cysteine (Cys), aspartic acid (Asp), glutamic acid (Glu), phenylalanine (Phe), glycine (Gly),
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`histidine (His), isoleucine (Ile), arginine (Arg), lysine (Lys), leucine (Leu), methionine (Met),
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`asparagine (Asn), proline (Pro), glutamine (Gln), serine (Ser), threonine (Thr), valine (Val),
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`tryptophan (Trp), tyrosine (Tyr), and combinations thereof. Stereoisomersof naturally-
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`occurring o-amino acids include, without limitation, D-alanine (D-Ala), D-cysteine (D-Cys),
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`D-aspartic acid (D-Asp), D-glutamic acid (D-Glu), D-phenylalanine (D-Phe), D-histidine
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`(D-His), D-isoleucine (D-Ile), D-arginine (D-Arg), D-lysine (D-Lys), D-leucine (D-Leu),
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`D-methionine (D-Met), D-asparagine (D-Asn), D-proline (D-Pro), D-glutamine (D-Gln),
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`D-serine (D-Ser), D-threonine (D-Thr), D-valine (D-Val), D-tryptophan (D-Trp), D-tyrosine
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`(D-Tyr), and combinations thereof.
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`Naturally-occurring aminoacids include those formed in proteins by post-translational
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`modification, such as citrulline (Cit).
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`Unnatural (non-naturally occurring) aminoacids include, without limitation, amino acid
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`analogs, amino acid mimetics, synthetic amino acids, N-substituted glycines, and N-methyl
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`amino acids in either the L- or D-configuration that function in a manner similarto the naturally-
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`occurring amino acids. For example, “amino acid analogs” can be unnatural aminoacidsthat
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`have the same basic chemical structure as naturally-occurring amino acids(i.e., a carbon thatis
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`bondedto a hydrogen, a carboxyl group, an amino group) but have modified side-chain groups
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`or modified peptide backbones, e.g., homoserine, norleucine, methionine sulfoxide, and
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`methionine methyl sulfonium. “Amino acid mimetics” refer to chemical compoundsthat have a
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`L7019.019PV1
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`structure that is different from the general chemical structure of an aminoacid, but that functions
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`in a mannersimilar to a naturally-occurring aminoacid.
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`“Linker” refers to a functional group that covalently bonds two or more moieties in a
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`compound or material. For example, the linking moiety can serve to covalently bond an
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`adjuvant moiety to an antibody construct in an immunoconjugate.
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`“Linking moiety” refers to a functional group that covalently bonds two or more moieties
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`in a compoundor material. For example, the linking moiety can serve to covalently bond an
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`adjuvant moiety to an antibody in an immunoconjugate. Useful bonds for connecting linking
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`moieties to proteins and other materials include, but are not limited to, amides, amines, esters,
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`carbamates, ureas, thioethers, thiocarbamates, thiocarbonates, and thioureas.
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`“Divalent” refers to a chemical moiety that contains two points of attachment for linking
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`two functional groups; polyvalent linking moieties can have additional points of attachment for
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`linking further functional groups. Divalent radicals may be denoted with the suffix “diyl”. For
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`example, divalent linking moieties include divalent polymer moieties such as divalent
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`poly(ethylene glycol), divalent cycloalkyl, divalent heterocycloalkyl, divalent aryl, and divalent
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`heteroaryl group. A “divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group” refers to a
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`cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group having two points of attachment for
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`covalently linking two moieties in a molecule or material. Cycloalkyl, heterocycloalkyl, aryl, or
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`heteroaryl groups can be substituted or unsubstituted. Cycloalkyl, heterocycloalkyl, aryl, or
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`heteroaryl groups can be substituted with one or more groups selected from halo, hydroxy,
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`amino, alkylamino, amido, acyl, nitro, cyano, and alkoxy.
`A wavyline (“ ot ”’) represents a point of attachment of the specified chemical moiety.
`If the specified chemical moiety has two wavylines (“ oh ”) present, it will be understoodthat
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`the chemical moiety can be usedbilaterally, i.e., as read from left to right or from rightto left.
`In some embodiments, a specified moiety having two wavy lines (“ st ”) present is considered
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`to be used as read from left to right.
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`“Alkyl”refers to a straight (linear) or branched, saturated, aliphatic radical having the
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`numberof carbon atoms indicated. Alkyl can include any numberof carbons, for example from
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`one to twelve. Examples of alkyl groups include, but are not limited to, methyl (Me, -CHs3), ethyl
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`(Et, -CH2CHs3), 1-propyl (n-Pr, n-propyl, -CH2CH2CHs), 2-propyl (i-Pr, i-propyl, -CH(CHs3)z), 1-
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`butyl (n-Bu, n-butyl, -CH2CH2CH2CHs), 2-methyl-1-propyl (i-Bu,1-butyl, -CH2CH(CHs3)2), 2-
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`butyl (s-Bu, s-butyl, -CH(CH3)CH2CHs), 2-methyl-2-propyl (t-Bu, t-butyl, -C(CH3)3), 1-pentyl
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`(n-pentyl, -CH2CH2CH2CH2CHs), 2-pentyl (-CH(CH3)CH2CH2CHs3), 3-pentyl (-CH(CH2CH3)2),
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`2-methyl-2-butyl (-C(CH3)2CH2CHs3), 3-methyl-2-butyl (-CH(CH3)CH(CHs3)2), 3-methyl-1-butyl
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`(-CH2CH2CH(CHs3)z), 2-methyl-1-butyl (-CH2CH(CH3)CH2CH3), 1-hexyl (-
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`CH2CH2CH2CH2CH?2CHs), 2-hexyl (-CH(CH3)CH2CH2CH2CHs3), 3-hexyl(-
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`CH(CH2CH3)(CH2CH2CH3)), 2-methyl-2-pentyl (-C(CH3)2CH2CH2CHs), 3-methyl-2-pentyl (-
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`CH(CH3)CH(CH3)CH2CH3), 4-methyl-2-pentyl (-CH(CH3)CH2CH(CHs3)z), 3-methyl-3-pentyl (-
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`C(CH3)(CH2CH3)2), 2-methyl-3-pentyl (-CH(CH2CH3)CH(CHs3)2), 2,3-dimethyl-2-butyl (-
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`C(CH3)2CH(CHs)z2), 3,3-dimethyl-2-butyl (-CH(CHs3)C(CHs)s, 1-heptyl, 1-octyl, and the like.
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`Alkyl groups can be substituted or unsubstituted. “Substituted alkyl” groups can be substituted
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`with one or more groups selected from halo, hydroxy, amino, oxo (=O), alkylamino, amido,
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`acyl, nitro, cyano, and alkoxy.
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`The term “alkyldiyl” refers to a divalent alkyl radical. Examples of alkyldiyl groups
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`include, but are not limited to, methylene (-CH2-), ethylene (-CH2CHb-), propylene(-
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`CH2CH2CH?-), and the like. An alkyldiyl group may also be referred to as an “alkylene” group.
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`“Alkenyl” refers to a straight (linear) or branched, unsaturated, aliphatic radical having
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`the numberof carbon atoms indicated and at least one carbon-carbon double bond, sp2. Alkenyl
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`can include from two to about 12 or more carbons atoms. Alkenyl groups are radicals having
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`“cis” and “trans” orientations, or alternatively, “E” and “Z”orientations. Examples include, but
`
`are not limited to, ethylenyl or vinyl (-CH=CH)2), allyl (-CH2CH=CHz2). butenyl, pentenyl, and
`
`isomers thereof. Alkenyl groups can be substituted or unsubstituted. “Substituted alkenyl”
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`groups can be substituted with one or more groups selected from halo, hydroxy, amino, oxo
`
`(=O), alkylamino, amido,acyl, nitro, cyano, and alkoxy.
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`The terms “alkenylene” or “alkenyldiyl”refer to a linear or branched-chain divalent
`
`hydrocarbon radical. Examples include, but are not limited to, ethylenylene or vinylene(-
`
`CH=CH-), allyl (-CH2CH=CH-), and thelike.
`
`“Alkynyl” refers to a straight (linear) or branched, unsaturated, aliphatic radical having
`
`the numberof carbon atoms indicated and at least one carbon-carbontriple bond, sp. Alkynyl
`
`25
`
`can include from two to about 12 or more carbons atoms. For example, C2-Ce alkynyl includes,
`
`but is not limited to ethynyl (-C=CH), propynyl (propargyl, -CH2C=CH), butynyl, pentynyl,
`
`hexynyl, and isomers thereof Alkynyl groups can be substituted or unsubstituted. “Substituted
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`alkynyl” groups can be substituted with one or more groups selected from halo, hydroxy, amino,
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`oxo (=O), alkylamino, amido, acyl, nitro, cyano, and alkoxy.
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`30
`
`The term “alkynylene” or “alkynyldiyl” refer to a divalent alkynyl radical.
`
`The terms “carbocycle”, “carbocyclyl”, “carbocyclic ring” and “cycloalkyl” refer to a
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`saturated or partially unsaturated, monocyclic, fused bicyclic, or bridged polycyclic ring
`
`assembly containing from 3 to 12 ring atoms, or the number of atoms indicated. Saturated
`
`monocyclic carbocyclic rings include, for example, cyclopropyl, cyclobutyl, cyclopentyl,
`
`35
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`cyclohexyl, and cyclooctyl. Saturated bicyclic and polycyclic carbocyclic rings include, for
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`L7019.019PV1
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`example, norbornane, [2.2.2] bicyclooctane, decahydronaphthalene and adamantane.
`
`Carbocyclic groups can also be partially unsaturated, having one or more double ortriple bonds
`
`in the ring. Representative carbocyclic groups that are partially unsaturated include, but are not
`
`limited to, cyclobutene, cyclopentene, cyclohexene, cyclohexadiene (1,3- and 1,4-isomers),
`
`cycloheptene, cycloheptadiene, cyclooctene, cyclooctadiene (1,3-, 1,4- and 1,5-isomers),
`
`norbornene, and norbornadiene.
`
`The term “cycloalkyldiyl” refers to a divalent cycloalkyl radical.
`
`“Aryl” refers to a monovalent aromatic hydrocarbon radical of 6-20 carbon atoms (C6—
`
`C20) derived by the removal of one hydrogen atom from a single carbon atom of a parent
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`10
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`aromatic ring system.. Aryl groups can be monocyclic, fused to form bicyclic or tricyclic
`
`groups, or linked by a bondto form a biaryl group. Representative aryl groups include phenyl,
`
`naphthyl and biphenyl. Otheraryl groups include benzyl, having a methylene linking group.
`
`Somearyl groups have from 6 to 12 ring members, such as phenyl, naphthyl or biphenyl. Other
`
`aryl groups have from 6 to 10 ring members, such as phenyl or naphthyl.
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`15
`
`The terms “arylene” or “aryldiyl” mean a divalent aromatic hydrocarbon radical of 6-20
`
`carbon atoms (Cs6—C20) derived by the removal of two hydrogen atom from a two carbon atoms
`
`of a parent aromatic ring system. Somearyldiyl groups are represented in the exemplary
`
`structures as “Ar”. Aryldiyl includes bicyclic radicals comprising an aromatic ring fused to a
`
`saturated, partially unsaturated ring, or aromatic carbocyclic ring. Typical aryldiyl groups
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`20
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`include, but are not limited to, radicals derived from benzene (phenyldiyl), substituted benzenes,
`
`naphthalene, anthracene, biphenylene, indenylene, indanylene, 1,2-dihydronaphthalene, 1,2,3,4-
`
`tetrahydronaphthyl, and the like. Aryldiyl groups are also referred to as “arylene”, and are
`
`optionally substituted with one or more substituents described herein.
`
`The terms “heterocycle,” “heterocyclyl” and “heterocyclic ring” are used
`
`25
`
`interchangeably herein and refer to a saturated or a partially unsaturated (i.e., having one or
`
`more double and/ortriple bonds within the ring) carbocyclic radical of 3 to about 20 ring atoms
`
`in whichat least one ring atom is a heteroatom selected from nitrogen, oxygen, phosphorus and
`
`sulfur, the remaining ring atoms being C, where one or more ring atoms is optionally substituted
`
`independently with one or more substituents described below. A heterocycle may bea
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`30
`
`monocycle having 3 to 7 ring members(2 to 6 carbon atoms and 1 to 4 heteroatoms selected
`
`from N, O, P, and S) or a bicycle having 7 to 10 ring members (4 to 9 carbon atoms and | to 6
`
`heteroatomsselected from N, O, P, and S), for example: a bicyclo [4,5], [5,5], [5,6], or [6,6]
`
`system. Heterocycles are described in Paquette, Leo A.; “Principles of Modern Heterocyclic
`
`Chemistry” (W.A. Benjamin, New York, 1968), particularly Chapters 1, 3, 4, 6, 7, and 9; “The
`
`35
`
`Chemistry of Heterocyclic Compounds, A series of Monographs” (John Wiley & Sons, New
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`L7019.019PV1
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`York, 1950 to present), in particular Volumes 13, 14, 16, 19, and 28; and J. Am. Chem. Soc.
`
`(1960) 82:5566. “Heterocyclyl”also includes radicals where heterocycle radicals are fused with
`
`a saturated, partially unsaturated ring, or aromatic carbocyclic or heterocyclic ring. Examples of
`
`heterocyclic rings include, but are not limited to, morpholin-4-yl, piperidin-1-yl, piperazinyl,
`
`piperazin-4-yl-2-one, piperazin-4-yl-3-one, pyrrolidin-1-yl, thiomorpholin-4-yl, S-
`
`dioxothiomorpholin-4-yl, azocan-1-yl, azetidin-1-yl, octahydropyrido[1,2-a]pyrazin-2-yl,
`
`[1,4]diazepan-1-yl, pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl,
`
`tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino,
`
`thiomorpholino, thioxanyl, piperazinyl, homopiperazinyl, azetidinyl, oxetanyl, thietanyl,
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`10
`
`homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl, diazepinyl, thiazepinyl, 2-pyrrolinyl, 3-
`
`pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl, dithianyl,
`
`dithiolanyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl, pyrazolidinylimidazolinyl,
`
`imidazolidinyl, 3-azabicyco[3.1.0]hexanyl, 3-azabicyclo[4.1.0]heptanyl,
`
`azabicyclo[2.2.2 |hexanyl, 3H-indolyl quinolizinyl and N-pyridyl ureas. Spiro heterocyclyl
`
`15
`
`moieties are also included within the scope of this definition. Examples of spiro heterocyclyl
`
`moieties include azaspiro[2.5]octanyl and azaspiro[2.4]heptanyl. Examples of a heterocyclic
`
`group wherein 2 ring atoms are substituted with oxo (~O) moieties are pyrimidinonyl] and 1,1-
`
`dioxo-thiomorpholinyl. The heterocycle groups herein are optionally substituted independently
`
`with one or more substituents described herein.
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`20
`
`The term “heterocyclyldiyl” refers to a divalent, saturated or a partially unsaturated (i.e.,
`
`having one or more double and/ortriple bonds within the ring) carbocyclic radical of 3 to about
`
`20 ring atoms in which at least one ring atom is a heteroatom selected from nitrogen, oxygen,
`
`phosphorusandsulfur, the remaining ring atoms being C, where one or morering atoms is
`
`optionally substituted independently with one or more substituents as described. Examples of 5-
`
`25
`
`membered and 6-membered heterocyclyldiyls include morpholinyldiyl, piperidinyldiyl,
`
`piperazinyldiyl, pyrrolidinyldiyl, dioxanyldiyl, thiomorpholinyldiyl, and S-
`
`dioxothiomorpholinyldiyl.
`
`The term “heteroaryl” refers to a monovalent aromatic radical of 5-, 6-, or 7-membered
`
`rings, and includes fused ring systems (at least one of which is aromatic) of 5-20 atoms,
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`30
`
`containing one or more heteroatoms independently selected from nitrogen, oxygen, and sulfur.
`
`Examples of heteroaryl groups are pyridinyl (including, for example, 2-hydroxypyridinyl),
`
`imidazolyl, imidazopyridinyl, pyrimidinyl (including, for example, 4-hydroxypyrimidinyl),
`
`pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxadiazolyl,
`
`oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinoliny|, tetrahydroisoquinolinyl, indolyl,
`
`35
`
`benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl,
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`L7019.019PV1
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`triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, thiadiazolyl, furazanyl,
`
`benzofurazanyl, benzothiophenyl, benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl,
`
`naphthyridinyl, and furopyridinyl. Heteroaryl groups are optiona