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WO2023178179A2 - Bi-functional molecules to degrade circulating proteins - Google Patents

Bi-functional molecules to degrade circulating proteins Download PDF

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Publication number
WO2023178179A2
WO2023178179A2 PCT/US2023/064436 US2023064436W WO2023178179A2 WO 2023178179 A2 WO2023178179 A2 WO 2023178179A2 US 2023064436 W US2023064436 W US 2023064436W WO 2023178179 A2 WO2023178179 A2 WO 2023178179A2
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Prior art keywords
compound
linker
mif
alkyl
formula
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PCT/US2023/064436
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WO2023178179A3 (en
Inventor
David Spiegel
David CAIANIELLO
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Yale University
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Yale University
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Priority claimed from US17/695,259 external-priority patent/US20230097887A1/en
Priority to AU2023234543A priority Critical patent/AU2023234543A1/en
Priority to CN202380040375.8A priority patent/CN119212726A/en
Priority to IL315581A priority patent/IL315581A/en
Priority to JP2024555118A priority patent/JP2025509736A/en
Priority to KR1020247033971A priority patent/KR20250004953A/en
Application filed by Yale University filed Critical Yale University
Priority to EP23771634.5A priority patent/EP4493220A2/en
Publication of WO2023178179A2 publication Critical patent/WO2023178179A2/en
Publication of WO2023178179A3 publication Critical patent/WO2023178179A3/en
Priority to MX2024011358A priority patent/MX2024011358A/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/55—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug, i.e. a dimer, oligomer or polymer of pharmacologically or therapeutically active compounds
    • 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
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    • A61P37/00—Drugs for immunological or allergic disorders
    • C—CHEMISTRY; METALLURGY
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    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
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    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
    • C07D493/02—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains two hetero rings
    • C07D493/08—Bridged systems
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    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
    • C07D493/02—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains two hetero rings
    • C07D493/10—Spiro-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
    • C07H15/02—Acyclic radicals, not substituted by cyclic structures
    • C07H15/04—Acyclic radicals, not substituted by cyclic structures attached to an oxygen atom of the saccharide radical
    • C07H15/08—Polyoxyalkylene derivatives
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
    • C07H15/02—Acyclic radicals, not substituted by cyclic structures
    • C07H15/04—Acyclic radicals, not substituted by cyclic structures attached to an oxygen atom of the saccharide radical
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    • C—CHEMISTRY; METALLURGY
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    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
    • C07H15/18—Acyclic radicals, substituted by carbocyclic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
    • C07H15/26—Acyclic or carbocyclic radicals, substituted by hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
    • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
    • C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
    • C07H19/056—Triazole or tetrazole radicals
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids

Definitions

  • MIF is a proinflammatory pluripotent cytokine which contributes to the development and perpetuation of many diseases. These include atherosclerosis, rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), sepsis, inflammatory bowel disease (IBD), among others.
  • RA rheumatoid arthritis
  • SLE systemic lupus erythematosus
  • IBD inflammatory bowel disease
  • MIF has been shown to participate in the progression of various cancers, including colon cancer, prostate cancer, breast cancer, lung cancer, cervical and adenocarcinoma, among others. In pre-clinical models, inhibition of MIF has been shown to attenuate symptoms and delay disease progression in the above-mentioned diseases.
  • MIF and IgG significantly contribute to the pathologic process of a number of diseases.
  • MIF neutralizing antibodies and small molecule MIF inhibitors have been developed, where IgG is upregulated in autoimmune and other diseases and causes untoward effects.
  • MIF acts through various pathways a MIF inhibitor is sometimes required to be able to interrupt multiple protein-protein interactions (PPIs), posing a challenge for drug development.
  • PPIs protein-protein interactions
  • IgG the removal of IgG molecules which are upregulated and contributing to autoimmune and/or other diseases is sometimes useful to inhibit and/or ameliorate the effects of the disease state. Therefore, a novel approach where MIF or IgG can be degraded as well as inhibited can overcome this difficulty and potentially provide a more robust therapeutic response.
  • the present inventors decided to further test them in clinical-relevant in vivo models and potentially develop them into therapeutics that target MIF or IgG in diseases. They also intend to use this basic bifunctional molecule strategy to target other circulating proteins of interest to degradation via the ASGPr.
  • the present disclosure is directed to bifunctional small molecules which can be used to inhibit and remove MIF or IgG.
  • the present disclosure aims to establish a general small molecule strategy to target the selective degradation of MIF or IgG, which contributes to the development and perpetuation of many diseases.
  • the bifunctional molecule construct contains a MIF binding motif derived from known small molecule ligands of MIF of a IgG binding motif which comprises a ligand which binds IgG. These moieties are referred to herein generically as macrophage migration inhibitory factor binding moiety (MIFBM) or alternatively, immunoglobulin G binding moiety (IgGBM).
  • MIFBM macrophage migration inhibitory factor binding moiety
  • IgGBM immunoglobulin G binding moiety
  • the other end of the bifunctional molecule is a motif that binds to hepatocyte asialoglycoprotein receptor (ASGPr), which is a triantennary N-acetylgalactosamine or other moiety as described herein.
  • ASGPr hepatocyte asialoglycoprotein receptor
  • ASGPRBM asialoglycoprotein receptor binding moiety
  • the motifs which are used to provide bifunctional compounds according to some embodiments, (MIFBM or IgGBM) and (ASGPRBM), are covalently linked via a linker such as a polyethylene glycol (PEG) or other linker as described herein with adjustable length or other linker as described herein, which optionally contains one or more connector molecule(s) which connects the linker to the MIFBM or IgGBM and/or the ASGPRBM.
  • a linker such as a polyethylene glycol (PEG) or other linker as described herein with adjustable length or other linker as described herein, which optionally contains one or more connector molecule(s) which connects the linker to the MIFBM or IgGBM and/or the ASGPRBM.
  • thje bifunctional compounds selectively bind to MIF or IgG in circulation and form a protein complex.
  • ASGPRBM asialoglycoprotein receptor binding moiety of the molecule
  • MIF or IgG is eliminated from circulation by hepatocytes, thus resulting in lowered levels of MIF or IgG with the result being that corresponding disease symptoms are attenuated and/or eliminated from a patient or subject administered the present compounds.
  • the MIF is substantially reduced or eliminated or the compromising IgG substantially reduced, resulting in substantially reduced symptoms or even a cure or elimination of the disease state or condition.
  • the approach pursuant to the present disclosure is inherently advantageous compared to the classical antibody -based strategy to target MIF or IgG of the prior art.
  • the small molecule based approach of the current disclosure overcomes the limitations of traditional antibody-based strategies, including lack of oral bioavailability, low-temperature storage requirements, immunogenicity, and high-cost.
  • the compounds and methods of the present disclosure are expected to have a more lasting effect compared to the conventional inhibitory approach because MIF or IgG is substantially reduced or eliminated by degradation inside hepatocytes rather than simply inhibited by reversibly blocking the protein-receptor interaction.
  • the bifunctional molecule construct pursuant to the present disclosure is also versatile in the sense that different disease states and/or conditions can be targeted by inhibiting and degrading MIF or IgG.
  • non-inhibitory protein binders presently of little value in therapy, can be potentially therapeutically useful in these small molecules.
  • the present disclosure is directed to compounds which are useful for removing circulating proteins which are associated with a disease state or condition in a patient or subject according to the general chemical structure: wherein [MIFBM/IgGBM] is a MIF or IgG binding moiety which binds respectively to circulating MIF or IgG, which are related to a disease state and/or condition and is to be removed by the action of hepatocytes on circulating protein (in some embodiments, the compounds selectively bind to MIF or IgG in plasma);
  • [ASGPRBM] is a binding moiety which binds to hepatocytes through asialoglycoprotein receptors which are on the surface of hepatocytes, such as in a patient or subject; each [CON] is an optional connector chemical moiety which, when present, connects directly to [MIFBM/IgGBM] or to [ASGPRBM] or connects the [LINKER] to [MIFBM/IgGBM] or to [ASGPRBM] and
  • [LINKER] is a chemical moiety having a valency ranging from 1 to 15, such as ranging from 1 to 10, ranging from 1 to 5 or 1, 2 or 3, which covalently attaches to one or more [ASGPRM] and/or [MIFBM/IgGBM] group, optionally through a [CON], including a [MULTICON] group, wherein said [LINKER] optionally itself contains one or more [CON] or [MULTICON] group(s); k’ is an integer ranging from 1 to 15, such as ranging from 1 to 10, ranging from 1 to 5, ranging from 1 to 3 or 1, 2 or 3; j’ is an integer ranging from 1 to 15, such as ranging from 1 to 10, ranging from 1 to 5, ranging from 1 to 3 or 1, 2 or 3; h and h’ are each independently an integer ranging from 0 to 15, such as ranging from 1 to 15, ranging from 1 to 10, ranging from 1 to 5, ranging from 1 to 3 or 1, 2 or 3, optionally h and/or h’
  • IL is an integer ranging from 0 to 15, such as ranging from 1 to 15, ranging from 1 to 10, ranging from 1 to 5, ranging from 1 to 3, or 1, 2 or 3, optionally 11 is an integer ranging from 1 to 5 or 1, 2 or 3 with the proviso that at least one of h, h’ and 11 is at least 1, or a pharmaceutically acceptable salt, stereoisomer, solvate or polymorph thereof.
  • [MIFBM] is a moiety according to the chemical structure: wherein XM is -(CH2)IM, -0-(CH2)IM, S-(CH2)IM, NRM-(CH2)IM, C(O)-(CH2)IM-, a polyethylene glycol (PEG) group containing from 1 to 8, such as 1-4 ethylene glycol residues or a -C(0)(CH2)IMNRM group;
  • RM is H or a C1-C3 alkyl group which is optionally substituted with one or two hydroxyl groups;
  • IM is 0-6, such as 1, 2, 3 or 4, such as 1.
  • [IgGMB] is a group according to the chemical structure: wherein DNP is a 2,4-dinitrophenyl group; or a group according to the chemical structure: wherein Y’ is H or NO2;
  • X is O, CH2, NR 1 , S(O), S(O) 2 , -S(O) 2 O, -OS(O) 2 , or OS(O) 2 O;
  • R 1 is H, a C1-C3 alkyl group, or a -C(O)(Ci-C3) group; or a group according to the chemical structure: wherein R 1 is the same as above; and K” is 1-5 (such as 3-5, such as 4), or a group represented by the chemical formula: wherein X’ is CH2, O, N-R 1 ’, or S;
  • R 1 is H or C1-C3 alkyl
  • Z is a bond, a monosaccharide, disaccharide, oligosaccharide, such as a sugar group selected from the monosaccharides, including aldoses and ketoses, and di saccharides, including those disaccharides described herein.
  • Monosaccharide aldoses include monosaccharides such as aldotriose (D-glyceraldehdye, among others), aldotetroses (D- erythrose and D-Threose, among others), aldopentoses, (D-ribose, D-arabinose, D-xylose, D- lyxose, among others), aldohexoses (D-allose, D-altrose, D-Glucose, D-Mannose, D-gulose, D-idose, D-galactose and D-Talose, among others), and the monosaccharide ketoses include monosaccharides such as ketotriose (dihydroxyacetone, among others), ketotetrose (D- erythrulose, among others), ketopentose (D-ribulose and D-xylulose, among others), ketohexoses (D-Psicone, D-Fructose, D-S
  • Exemplary disaccharides which find use in the present disclosure include sucrose (which may have the glucose optionally N-acetylated), lactose (which may have the galactose and/or the glucose optionally N-acetylated), maltose (which may have one or both of the glucose residues optionally N-acetylated), trehalose (which may have one or both of the glucose residues optionally N-acetylated), cellobiose (which may have one or both of the glucose residues optionally N-acetylated), kojibiose (which may have one or both of the glucose residues optionally N-acetylated), nigerose (which may have one or both of the glucose residues optionally N-acetylated), isomaltose (which may have one or both of the glucose residues optionally N-acetylated), P,P-trehalose (which may have one or both of the glucose residues optionally N-acetylated), sophorose
  • [IgGBM] is a group according to the chemical structure: a-L-Rhamnose p-L-Rhamnose Phosphoryl Choline
  • R 1 is H or a C1-C3 alkyl group.
  • [IgGBM] is a group according to the chemical structure: , wherein X b is a bond, O, CH2 or NR 1 or S; and
  • R 1 is the same as above; or a group according to the chemical structure: where R N02 is a dinitrophenyl group optionally linked through CH2, S(O), S(O)2, - S(O) 2 O, -OS(O) 2 , or OS(O) 2 O; or a dinitrophenyl group according to the chemical structure: where K’” is 1-4 (such as 2-3, such as 3), or a a group according to a chemical structure which is set forth in FIGURE 67 hereof which is covalently attached to a [CON] group, a [LINKER] group or a [ASGPRBM] group through an amine group, such as a primary or secondary alkyl amine group which is optionally substituted on the amine group with a C1-C3 alkyl group.
  • [IgGBM] is a peptide according to the sequence (all references cited are incorporated by reference herein):
  • D-PAM (Verdoliva, et al., J. Immunol. Methods, 2002, 271, 77-88);
  • FGRLVSSIRY (Krook, et al., J. Immunol. Methods 1998, 221, 151-157) SEQ ID NO:2;
  • FcBP-2 (Dias, et al., J. Am. Chem. Soc. 2006, 128, 2726-2732);
  • FcRM Fc Receptor Mimetic, Verdoliva, et al., ChemBioChem 2005, 6, 1242-1253
  • HWRGWV (Yang, et al., J. Peptide Res. 2006, 66, 110-137) SEQ ID NO:5;
  • HYFKFD (Yang, et al., J. Chromatogr. A 2009, 1216, 910-918) SEQ ID NO: 6;
  • HFRRHL (Menegatti, et al., J. Chromatogr. A 2016, 1445, 93-104) SEQ IDNO:7;
  • HWCitGWV (Menegatti, et al., J. Chromatogr. A 2016, 1445, 93-104)
  • D2AAG Mall Synthetic peptide ligand, Lund, et al., J. Chromatogr. A 2012, 1225, 158-167;
  • DAAG Mall Synthetic peptide ligand, Lund, et al., J. Chromatogr. A 2012, 1225, 158-167
  • cyclo[(N-Ac)S(A)-RWHYFK-Lact-E] (Menegatti, et al., 4 //a/. Chem. 2013, 85, 9229-9237)
  • NKFRGKYK (Sugita, et al., Biochem. Eng. J. 2013, 79, 33-40) SEQ ID NO: 12;
  • NARKFYKG (Sugita, et al., Biochem. Eng. J. 2013, 79, 33-40) SEQ ID NO: 13;
  • FYCHWALE (Zhao, et al., J. Chromatogr. A 2014, 1355, 107-114) SEQ ID NO: 15;
  • FYCHTIDE (Zhao, et al., J. Chromatogr. A 2014, 1359, 100-111) SEQ ID NO: 16;
  • RRGW (Tsai, et al., Anal. Chem. 2014, 86, 2931-2938) SEQ ID NO: 17;
  • KHRFNKD (Yoo and Choi, BioChip J. 2015, 10, 88-94) SEQ ID NO: 18;
  • [ASGPRBM] is a group according to the chemical structure: where X is 1-4 atoms in length and comprises O, S, N(R N1 ) or C(R N1 )(R N1 ) groups such that when X is 1 atom in length, X is O, S, N(R N1 ) or C(R N1 )(R N1 ), when X is 2 atoms in length, no more than 1 atom of X is O, S or N(R N1 ), when X is 3 or 4 atoms in length, no more than 2 atoms of X are O, S or N(R N1 ); where R N1 is H or a C1-C3 alkyl group optionally substituted with from 1-3 halo groups, such as F (in some embodiments, R N1 is H or methyl);
  • Ri and R3 are each independently H, -(CH2)KOH, -(CH2)KOCI-C4 alkyl, which is optionally substituted with from 1-3 halo (F, Cl, Br, or I) groups, C1-C4 alkyl, which is optionally substituted with from 1-3 halo (F, Cl, Br, or I) groups,
  • Ri and R3 are each independently a group, which is optionally substituted with up to three (such as 1) halo groups (such as F), C1-C4 alkyl groups, each of which is optionally substituted with from one to three halo groups, such as F, or one or two hydroxyl groups, or O-C1-C4 alkyl groups, each of which alkyl groups is optionally substituted with from one to three halo groups, such as F, or one or two hydroxyl groups, and
  • K is independently an integer ranging from 0-4 (such as 0, 1, 2, 3 or 4), or
  • Ri and R3 are each independently a group according to the chemical structure:
  • R 7 is O-C1-C4 alkyl, which is optionally substituted with from 1 to 3 halo groups, such as F and 1 or 2 hydroxy groups, or R 7 is a
  • -NR“R“ group or Ri and R3 are each independently a group according to the structure: wherein CYC is a ring selected from the group consisting of: saturated carbocyclic, wherein each of LINKERX, R c , and -(CH2)K- are attached to an open valence in CYC, including N-H;
  • R c is absent, H, C1-C4 alkyl which is optionally substituted with from 1-3 halo (such as F) groups or 1-2 hydroxyl groups, or a group according to the structure: where R4, Rs and Re are each independently, H, halo (F, Cl, Br, I), CN, NR N1 R N2 , -(CH2)KOH, -(CH 2 )KOCI-C 4 alkyl, which is optionally substituted with from 1-3 halo (F, Cl, Br, I) groups, C1-C3 alkyl, which is optionally substituted with from 1-3 halo (F, Cl, Br, I) groups, -O-Ci-C3-alkyl, which is optionally substituted with from 1-3 halo, such as F groups, -(CH2)KC00H, -(CH2)KC(O)O-CI-C4 alkyl which is optionally substituted with from 1-3 halo, such as F groups, O-C(O)
  • K is independently an integer ranging from 0-4 (0, 1, 2, 3 or 4);
  • K’ is an integer ranging from 1-4, such as 1;
  • R N3 is H, or a C1-C3 alkyl group which is optionally substituted with 1-3 halo groups, such as F or 1 or 2 hydroxy groups;
  • R N4 is H, a C1-C3 alkyl group which is optionally substituted with 1-3 halo groups, such as F or 1 or 2 hydroxy groups, or R N4 is a group, where K is sometimes 1;
  • LJNKERX is a linker group which is comprises to at least one [MIFBM/IgGBM] group and links the [MIFBM/IgGBM] group to the [ASGPRBM] through one or more optional [CON] groups, or
  • I - i is a linker group which contains at least one or more functional groups which can be used to covalently bond the linker group to at least one [MIFBM/IgGBM] group or optional [CON] group; group where R and K are the same as above;
  • R AM is H, a C1-C4 alkyl group optionally substituted with up to 3 halo groups (such as F) and one or two hydroxyl groups, a -(CH2)KCOOH group, a -(CH2)KC(O)O-CI-C4 alkyl group which is optionally substituted with from 1-3 halo, such as F groups, a
  • O-C(O)-Ci-C4 alkyl group which is optionally substituted with from 1-3 halo, such as F groups, a -C(O)-Ci-C4 alkyl group, which is optionally substituted with from 1-3 halo, such as F groups, a -(CH2)K-NR N3 R N4 group where R N3 is H, or a C1-C3 alkyl group which is optionally substituted with 1-3 halo groups, such as F or 1 or 2 hydroxy groups; and
  • R N4 is H, a C1-C3 alkyl group which is optionally substituted with 1-3 halo groups, such as F or 1 or 2 hydroxy groups, or R N4 is a group (K is sometimes
  • R TA is H, CN, NR N1 R N2 , -(CH2)KOH, -(CH2)KOCI-C4 alkyl, which is optionally substituted with from 1-3 halo (F, Cl, Br, or I) groups, C1-C4 alkyl, which is optionally substituted with from 1-3 halo (F, Cl, Br, or I) groups, -(CH2)KCOOH, -(CH2)KC(O)O-CI-C4 alkyl which is optionally substituted with from 1-3 halo, such as F groups, O-C(O)-Ci-C4 alkyl, which is optionally substituted with from 1-3 halo, such as F groups, -C(O)-Ci-C4 alkyl, which is optionally substituted with from 1-3 halo, such as F groups, or R TA is a C3-C10 aryl or a three- to ten-membered heteroaryl group containing up
  • R TA is or
  • R TA is a group which is optionally substituted with up to three, such as
  • halo such as F
  • R N , R N1 and R N2 are each independently H or a C1-C3 alkyl group which is optionally substituted with from one to three halo groups, such as F, or one or two hydroxyl groups and wherein each -(CH2)K group is optionally substituted with 1-4, such as 1 or 2, C1-C3
  • [CON] is a connector moiety (including a [MULTICON]) as otherwise described herein.
  • [LINKER] is a linking moiety as otherwise described herein which links [MIFBM/IgGBM] to the [ASGPRBM] group and optionally contains one or more connector moieties (which optionally connect(s) more than one chemical moiety to provide said linking moiety or which connects said linking moiety to said [MIFBM/IgGBM] group or said [ASGPRBM] group, or a pharmaceutically acceptable salt, stereoisomer, solvate or polymorph thereof.
  • X is -O-C(R N1 )(R N1 ),
  • X is -O-C(R N1 )(R N1 )-C(R N1 )(R N1 ), C(R N1 )(R N1 )-O-C(R N1 )(R N1 )-, -O-C(R N1 )(R N1 )-O-, -O-C(R N1 )(R N1 )-S-, -O-C(R N1 )(R N1 )-N(R N1 )-, -S-C(R N1 )(R N1 )-C(R N1 )(R N1 ), C(R N1 )(R N1 )-S-C(R N1 )(R N1 )-, C(R N1 )(R N1 )-C(R N1 )(R N1 )-S, -S-C(R N1 )(R N1 )-S-, -S-C(R
  • X is OCH2 or CH2O.
  • R N1 is H.
  • the [ASGPRBM] group is a group according to the chemical structure: where Ri, R2 and R3 are the same as above, or a pharmaceutically acceptable salt, stereoisomer, solvate or polymorph thereof.
  • the [ASGPRBM] group is a group according to the chemical structure: where R A is a C1-C3 alkyl group which is optionally substituted with 1-5 halo (such as F) groups (In some embodiments, R A is a methyl or ethyl group which is optionally substituted with from 1-3 fluoro groups);
  • ZA is -(CH 2 )IM, -O-(CH 2 )IM, S-(CH 2 )IM, NRM-(CH 2 )IM, C(O)-(CH 2 )IM-, a PEG group containing from 1 to 8, such as 1-4 ethylene glycol residues or a -C(0)(CH2)IMNRM group (In some embodiments, a PEG containing group comprising from 1 to 8 ethylene glycol, such as 2-4 ethylene glycol residues) where IM and RM are the same as above; and
  • ZB is absent, (CH2)IM, C(0)-(CH2)IM- or C(0)-(CH2)IM-NRM, where IM and RM are the same as above.
  • Ri and R3 are each independently often a group according to the chemical structure: are the same as above.
  • the compounds include the compounds which are presented in the above-described compounds. In some embodiments, in the above-described compounds, in some embodiments, the compounds include the compounds which are presented in
  • FIGURES 1, 7 and 13 additional compounds are presented in FIGURES 16-66 and include final compounds set forth therein and intermediates which are used to make final compounds pursuant to the present disclosure.
  • Ri and R3 of the [ASGPRBM] group include those moieties which are presented in FIGURE 68 hereof.
  • R2 of the [ASGPRBM] group include those moieties which are presented in FIGURE 69 hereof.
  • the IgGBM group is a FCIII group according to the chemical or a FcIII-4c peptide represented . (SEQ ID NO:20).
  • the present disclosure is directed to a pharmaceutical composition
  • a pharmaceutical composition comprising an effective amount of a compound according to the present disclosure in combination with a pharmaceutically acceptable carrier, additive or excipient, optionally in combination with at least one additional bioactive agent.
  • the present disclosure is directed to a method of treating a disease state or condition where MIF is related to or contributes to a disease state and or condiiton or the symptomology associated with the disease state or condition.
  • MIF rheumatoid arthritis
  • SLE systemic lupus erythematosus
  • Alzheimer’s disease atherosclerosis
  • heart disease stroke and cancer (including leukemia), among numerous others.
  • the method of treatment comprises administering to a patient or subject in need of therapy an effective amount of at least one compound according to the present disclosure, optionally in combination with an additional bioactive agent to reduce the likelihood of, inhibit and/or treat the disease state or condition by removing MIF associated with the disease state and/or condition from the circulation of the patient or subject.
  • MIF-GN3 were able to induce the degradation of human MIF injected in mice. Additional work focuses on the evaluation of MIF-GN3 in autoimmune disease models as well as seeking to optimize the bifunctional MIF degrader using different MIF and ASGPr ligands. IgG-targeting molecules were able to recapitulate our results with MIF in cell culture and mediated the removal of IgG from mouse serum.
  • the present disclosure provides a useful platform for the development of new therapeutics for these critical diseases.
  • FIGURE 1 shows representative compounds according to some embodiments. Note that the figure discloses compound 3w (negative control for MIF inhibition), MIF-NVS- PEGnGN3, MIFGN3, MIF-PEGnGN3, MIF-AcF3-l, MIF-AcF3-2 and MIF-AcF3-3. Note that n in the PEG linker sometimes ranges from 1-12, such as 1 to 10, 2 to 8, 2 to 6, 2 to 5 or 1, 2, 3 or 4.
  • FIGURE 2 shows fluorescence polarization data of MIF-FITC binding to human MIF, indicating that our MIF -binding moiety binds MIF, in accordance with some embodiments.
  • Bifunctional molecules WJ-PEG4-GN3, WJ-PEG2-GN3, and NVS-PEG3- GN3 bound competitively with MIF-FITC, indicating that the bifunctional molecules maintain the ability to bind human MIF.
  • FIGURE 3 shows that bifunctional molecules are able to deplete human MIF from the supernatant of culture HepG2 cells, in accordance with some embodiments.
  • FIGURE 4 shows that MIF internalized by HepG2 cells is trafficked to lysosomes, in accordance with some embodiments.
  • FIGURE 5 shows that MIF-GN3 mediates the depletion of injected human MIF from mice, in accordance with some embodiments.
  • FIGURE 6 shows that MIF-GN3 is able to delay tumor growth in a mouse model of prostate cancer, in accordance with some embodiments.
  • FIGURE 7 shows molecules DNP-GN3 and DNP-AcF3-3, which are bifunctional moelcules that bind to anti-DNP IgG and ASGPR, in accordance with some embodiments.
  • FIGURE 8 shows that DNP-GN3 and DNP-AcF3-3 mediate the formation of a ternary complex between HepG2 cells and anti-DNP, in accordance with some embodiments.
  • FIGURE 9 shows that DNP-GN3 and DNP-AcF3-3 mediate the uptake of alexa 488- labeled anti-DNP by HepG2 cells, in accordance with some embodiments.
  • FIGURE 10 shows that DNP-GN3 and DNP-AcF3-3 mediate the localization of alexa 568 labeled anti-DNP to late endosomes and lysosomes, in accordance with some embodiments.
  • FIGURE 11 shows that DNP-AcF3-3 mediates the degradation of alexa 488-labeled anti-DNP in HepG2 cells, in accordance with some embodiments.
  • FIGURE 12 shows that DNP-GN3 mediates the depletion of anti-DNP from mouse serum, in accordance with some embodiments.
  • FIGURE 13 shows the structures of IgG-degrading molecules IBA-GN3, Triazine- GN3, FcIII-GN3, and FcIII-4c-GN3, in accordance with some embodiments.
  • FIGURE 14 shows that FcIII-GN3 mediates the uptake of human IgG into HepG2 cells, in accordance with some embodiments. Experiment performed as described above.
  • FIGURE 15 shows that FcIII-GN3 mediates the localization of IgG to late endosomes in HepG2 cells, in accordance with some embodiments. Experiment performed as described above.
  • FIGURES 16-18 show the synthesis of PEG linkers used in several molecules , in accordance with some embodiments.
  • FIGURES 19-21 show the synthesis of ASGPR-binding precursors and ligands used in several molecules , in accordance with some embodiments.
  • FIGURES 22-26 show the synthesis of valency linkers used in several molecules , in accordance with some embodiments.
  • FIGURES 27-28 show the synthesis of MIF ligands used in several bifunctional molecules, in accordance with some embodiments.
  • FIGURE 29 describes the synthesis of the bifunctional molecule MIF-NVS-PEGn- GN3, in accordance with some embodiments.
  • FIGURE 30 describes the synthesis of bifunctional molecules MIF-GN3 and MIF- PEGn-GN3, in accordance with some embodiments.
  • FIGURE 31 describes the synthesis of the bifunctional molecule targeting MIF and ASGPR, containing one bicyclic ASGPR AcF3 ligands, in accordance with some embodiments.
  • FIGURE 32 describes the synthesis of the bifunctional molecule targeting MIF and ASGPR, containing two bicyclic ASGPr AcF3 ligands, in accordance with some embodiments.
  • FIGURE 33 describes the synthesis of the bifunctional molecule targeting MIF and ASGPR, containing three bicyclic ASGPr ligands, in accordance with some embodiments.
  • FIGURE 34 shows the synthesis of DNP-GN3, in accordance with some embodiments.
  • FIGURE 35 shows the synthesis of DNP-AcF3-3, in accordance with some embodiments.
  • FIGURE 36 shows the synthetic scheme used to obtain IBA-GN3, in accordance with some embodiments.
  • FIGURE 37 shows the synthesis of triazine-GN3, in accordance with some embodiments.
  • FIGURE 38 shows the synthetic scheme used to access FcIII-GN3, in accordance with some embodiments.
  • FIGURE 39 shows the synthetic scheme used to access FcIII-4c-GN3, in accordance with some embodiments.
  • FIGURES 40-43 describe the synthesis of bifunctional molecules targeting MIF and ASGPr, containing three bicyclic ASGPR ligands with different substitutions on the 2-amine of the sugar, in accordance with some embodiments.
  • FIGURE 44 shows the synthesis of compound MIF- 18-3, in accordance with some embodiments.
  • FIGURE 45 shows the synthesis of compound MIF-31-3, in accordance with some embodiments.
  • FIGURE 46 shows the synthesis of compound MIF-15-3, in accordance with some embodiments.
  • FIGURE 47 shows the synthesis of compound MIF- 19-3, in accordance with some embodiments.
  • FIGURE 48 shows the synthesis of compound MIF- 16-3, in accordance with some embodiments.
  • FIGURE 49 shows the synthesis of compound MIF-20-3, in accordance with some embodiments.
  • FIGURE 50 shows the synthesis of compound MIF- 14-3, in accordance with some embodiments.
  • FIGURE 51 shows the synthesis of compound MIF-21-3, in accordance with some embodiments.
  • FIGURES 52-66 show the synthesis of a number of MIF-binding compounds with various ASGPRBM moieties, in accordance with some embodiments.
  • FIGURE 67 shows exemplary IgGBM groups each of which is covalently attached to a [CON] group, a [LINKER] group or a [ASGPRBM] group through an amine group, such as a primary or secondary alkyl amine group which is optionally substituted on the amine group with a C1-C3 alkyl group, in accordance with some embodiments.
  • FIGURE 68 shows exemplary Ri and R3 substituents on ASGPRBM groups, in accordance with some embodiments.
  • FIGURE 69 shows exemplary R2 substituents on ASGPRBM groups, in accordance with some embodiments.
  • the present disclosure provides bi-functional molecules that utilize the endo-lysosomal machinery in the liver to eliminate MIF and/or IgG.
  • the present disclosure utilizes bi-functional molecules to target MIF or IgG and is modular and versatile. This offers convenience to apply medicinal chemistry to optimize the molecules. For example, in preliminary experiments, incorporation of more optimized ASGPr binding motifs have been shown to enhance MIF or IgG degradation in vitro. Since the bi-functional molecule is capable of degrading MIF or IgG, it does not require the MIF- binding motif (MIFBM) or the IgG binding moiety (IgGBM)to be an inhibitor, but in practice many of these binders will function as inhibitors of MIF or IgG.
  • MIFBM MIF- binding motif
  • IgGBM IgG binding moiety
  • the present disclosure is directed to bi-functional compounds which find use as pharmaceutical agents in the treatment of disease states and/or conditions which are mediated through macrophage migration inhibitory factor (MIF) or Immunoglobulin G (IgG).
  • MIF macrophage migration inhibitory factor
  • IgG Immunoglobulin G
  • the present disclosure is directed to pharmaceutical compositions which comprise these bi-functional compounds as well as methods for treating disease states and/or conditions which are mediated through MIF or IgG or where MIF and IgG are contributing factors to the development and perpetuation of diseases and/or conditions, especially including autoimmune diseases and cancer, among others.
  • present disclosure provides a molecular strategy to lower plasma MIF level or IgG levels in patients with autoimmune diseases or certain types of cancers and other diseases.
  • the bi- functional molecule construct is comprised of a MIF-targeting motif that is derived from small molecule MIF ligands or an IgG-binding motif which binds to IgG, and an ASGPr- targeting motif that binds to hepatocyte asialoglycoprotein receptor (ASGPr).
  • the compounds selectively bind MIF or IgG in plasma and subsequently engage the endo- lysosomal pathway of hepatocytes through ASGPr.
  • MIF or IgG is internalized and degraded by hepatocytes, thus resulting in potential attenuation of corresponding disease symptoms which are modulated through MIF and/or IgG.
  • compound refers to any specific chemical compound disclosed herein and includes tautomers, regioisomers, geometric isomers, stereoisomers and where applicable, optical isomers (enantiomers) thereof, as well as pharmaceutically acceptable salts and derivatives (including prodrug forms) thereof.
  • compound generally refers to a single compound, but also may include other compounds such as stereoisomers, regioisomers and/or optical isomers (including racemic mixtures) as well as specific enantiomers or enantiomerically enriched mixtures of disclosed compounds.
  • the term also refers, within context, to prodrug forms of compounds which have been modified to facilitate the administration and delivery of compounds to a site of activity. It is noted that in describing the present compounds, numerous substituents, linkers and connector molecules and variables associated with same, among others, are described.
  • the use of a bond presented as - signifies that a single bond is present or absent, depending on the context of the chemistry described, including the attachment of the bond to another moiety.
  • the use of a bond presented as - signifies that a single bond or a double bond is intended depending on the context of the chemistry described. It is understood by those of ordinary skill that molecules which are described herein are stable compounds as generally described hereunder.
  • patient or “subject” is used throughout the specification within context to describe an animal, generally a mammal, such as a human, to whom treatment, including prophylactic treatment (prophylaxis, including especially as that term is used with respect to reducing the likelihood of metastasis of an existing cancer), with the compositions according to the present disclosure is provided.
  • treatment including prophylactic treatment (prophylaxis, including especially as that term is used with respect to reducing the likelihood of metastasis of an existing cancer)
  • prophylactic treatment prophylaxis, including especially as that term is used with respect to reducing the likelihood of metastasis of an existing cancer
  • Compounds according to the present disclosure are useful for the treatment of numerous disease states including autoimmune diseease states and/or conditions and inflammatory disease states and/or conditions as well as cancer, including especially for use in reducing the likelihood of metastasis or recurrence of a cancer.
  • a disease state e.g. an autoimmune disease such as rheumatoid arthritis (RA) or systemic lupus erythematosus (SLE), among others, atherosclerosis, heart disease or stroke, among numerous others or a cancer, including leukemia
  • RA rheumatoid arthritis
  • SLE systemic lupus erythematosus
  • a disease state or condition for which a MIF protein may be removed refers to any action providing a benefit to a patient at risk for a disease state or condition for which a MIF protein may be removed, such as an autoimmune disease including rheumatoid arthritis (RA) or systemic lupus erythematosus (SLE), among others, atherosclerosis, heart disease, stroke and cancer (including leukemia) including recurrence and/or metastasis of cancer, improvement in the condition through lessening or suppression of at least one symptom of the disease state or condition, inhibition of one or more manifestations of the disease state (e.g., plaque formation, heart disease, cancer growth, reduction in cancer cells or tissue), prevention, reduction in the likelihood or delay in progression of a disease state or condition or manifestation of the disease state or condition, especially including plaque formation in atheroslerosis, deterioration of tissue and inflammation in rheumatoid arthritis, further damage to cardiovascular tissue in heart disease, further damage to central nervous tissue
  • RA
  • Treatment encompasses both prophylactic and therapeutic treatment, depending on the context of the treatment.
  • prophylactic when used, means to reduce the likelihood of an occurrence or the severity of an occurrence within the context of treatment of disease state or condition, as otherwise described hereinabove.
  • MIFBM macrophage migration inhibitory factor binding moiety
  • ASGPRBM asialoglycoprotein receptor binding moiety
  • the bifunctional molecule to which is bound circulating protein is internalized by the hepatocyte through an endocytosis mechanism whereupon the cell will destroy the protein via a lysosomal degradation or other degradation pathway.
  • the term “immunoglobulin G binding moiety” or “IgGBM” is used to describe a moiety which binds to circulating IgG immunoglobulin, forming a complex with bifunctional molecules according to the present disclosure to be ultimately destroyed in hepatocytes.
  • the terms MIFBM and IgGBM are used synonymously.
  • MIFBMs for inclusion in bifunctional compounds according to the present disclosure include moieties found in bifunctional chemical structures which appear in Figure 1, attached hereto.
  • MIFBMs include moieties according to the wherein XM is -(CH 2 )IM, -0-(CH 2 )IM, S-(CH 2 )IM, NRM-(CH 2 )IM, C(O)-(CH 2 )IM-, a PEG group containing from 1 to 8, such as 1-4 ethylene glycol residues or a -C(O)(CH 2 )IMNRM group;
  • RM is H or a C1-C3 alkyl group which is optionally substituted with one or two hydroxyl groups;
  • IM is 0-6, such as 1, 2, 3 or 4.
  • AGPRBM asialoglycoprotein receptor binding moiety
  • This binding moiety is also a component of the presently claimed bifunctional compounds bound to the MIHBM moiety through a linker.
  • the ASGPRBM selectively binds to hepatocyte asialoglycoprotein receptor on the surface of hepatocytes. It is through this moiety that bifunctional compounds complexed with circulating protein bind to hepatocytes. Once bound to the hepatocyte, the circulating MIF protein is taken into the hepatocytes via a phagocytosis mechanism wherein the circulating protein is degraded through lysosomal degradation.
  • Exemplary ASGPRBM groups for use in compounds according to the present disclosure include moieties according to the chemical structures: where X is 1-4 atoms in length and comprises O, S, N(R N1 ) or C(R N1 )(R N1 ) groups such that when X is 1 atom in length, X is O, S, N(R N1 ) or C(R N1 )(R N1 ), when X is 2 atoms in length, no more than 1 atom of X is O, S or N(R N1 ), when X is 3 or 4 atoms in length, no more than 2 atoms of X are O, S or N(R N1 ); where R N1 is H or a C1-C3 alkyl group optionally substituted with from 1-3 halo groups, such as F (in some embodiments, R N1 is H or methyl); R 1 and R 3 are each independently H, -(CH 2 ) K OH, -(CH 2 ) K
  • [CON] is a connector moiety (including a [MULTICON]) as otherwise described herein.
  • [LINKER] is a linking moiety as otherwise described herein which links [MIFBM/IgGBM] to the [ASGPRBM] group and optionally contains one or more connector moieties (which optionally connect(s) more than one chemical moiety to provide said linking moiety or which connects said linking moiety to said [MIFBM/IgGBM] group or said [ASGPRBM] group, or a pharmaceutically acceptable salt, stereoisomer, solvate or polymorph thereof.
  • X is -O-C(R N1 )(R N1 ),
  • X is -O-C(R N1 )(R N1 )-C(R N1 )(R N1 ), C(R N1 )(R N1 )-O-C(R N1 )(R N1 )-, -O-C(R N1 )(R N1 )-O-, -O-C(R N1 )(R N1 )-S-, -O-C(R N1 )(R N1 )-N(R N1 )-, -S-C(R N1 )(R N1 )-C(R N1 )(R N1 ), C(R N1 )(R N1 )-S-C(R N1 )(R N1 )-, C(R N1 )(R N1 )-C(R N1 )(R N1 )-S, -S-C(R N1 )(R N1 )-S-, -S-C(R
  • X is -O-C(R N1 )(R N1 )-C(R N1 )(R N1 )-C(R N1 )(R N1 ), C(R N1 )(R N1 )-O-C(R N1 )(R N1 )- (R N1 )(R N1 )_, -O-C(R N1 )(R N1 )-O-C(R N1 )(R N1 )-, -S-C(R N1 )(R N1 )-C(R N1 )(R N1 )- C(R N1 )(R N1 )-, C(R N1 )(R N1 )-S-C(R N1 )(R N1 )-C(R N1 )(R N1 )-, C(R N1 )(R N1 )-(R N1 )-S-C(R N1 )(R N1
  • X is OCH2 or CH2O.
  • R N1 is H.
  • the [ASGPRBM] group is a group according to the chemical structure: where R1, R2 and R3 are the same as above, or a pharmaceutically acceptable salt, stereoisomer, solvate or polymorph thereof.
  • the [ASGPRBM] group is a group according to the chemical structure: where R A is a C 1 -C 3 alkyl group which is optionally substituted with 1-5 halo (such as fluoro) groups (in some embodiments, R A is a methyl or ethyl group which is optionally substituted with from 1-3 fluoro groups); Z A is -(CH 2 ) IM, -O-(CH 2 ) IM , S-(CH 2 ) IM , NR M -(CH 2 ) IM, C(O)-(CH 2 ) IM - , a polyethylene glycol (PEG) group containing from 1 to 8, such as 1-4 ethylene glycol residues or a - C(O)(CH2)
  • neoplasia or “cancer” is used throughout the specification to refer to the pathological process that results in the formation and growth of a cancerous or malignant neoplasm, i.e., abnormal tissue that grows by cellular proliferation, often more rapidly than normal and continues to grow after the stimuli that initiated the new growth cease.
  • malignant neoplasms show partial or complete lack of structural organization and functional coordination with the normal tissue and most invade surrounding tissues, metastasize to several sites, and are likely to recur after attempted removal and to cause the death of the patient unless adequately treated.
  • Neoplasms include, without limitation, morphological irregularities in cells in tissue of a subject or host, as well as pathologic proliferation of cells in tissue of a subject, as compared with normal proliferation in the same type of tissue. Additionally, neoplasms include benign tumors and malignant tumors (e.g., colon tumors) that are either invasive or noninvasive.
  • Malignant neoplasms are distinguished from benign neoplasms in that the former show a greater degree of anaplasia, or loss of differentiation and orientation of cells, and have the properties of invasion and metastasis.
  • neoplasms or neoplasias from which the target cell of the present disclosure may be derived include, without limitation, carcinomas (e.g., squamous-cell carcinomas, adenocarcinomas, hepatocellular carcinomas, and renal cell carcinomas), particularly those of the bladder, bowel, breast, cervix, colon, esophagus, head, kidney, liver, lung, neck, ovary, pancreas, prostate, and stomach; leukemias; benign and malignant lymphomas, particularly Burkitt's lymphoma and Non-Hodgkin's lymphoma; benign and malignant melanomas; myeloproliferative diseases; sarcomas, particularly Ewing's sarcoma
  • Representative common cancers to be treated with compounds according to the present disclosure include, for example, prostate cancer, metastatic prostate cancer, stomach, colon, rectal, liver, pancreatic, lung, breast, cervix uteri, corpus uteri, ovary, testis, bladder, renal, brain/CNS, head and neck, throat, Hodgkin’s disease, non-Hodgkin’s lymphoma, multiple myeloma, leukemia, melanoma, non-melanoma skin cancer, acute lymphocytic leukemia, acute myelogenous leukemia, Ewing’s sarcoma, small cell lung cancer, choriocarcinoma, rhabdomyosarcoma, Wilms’ tumor, neuroblastoma, hairy cell leukemia, mouth/pharynx, oesophagus, larynx, kidney cancer and lymphoma, among others, which may be treated by one or more compounds according to the present disclosure.
  • the present disclosure has general applicability treating virtually any cancer in any tissue, thus the compounds, compositions and methods of the present disclosure are generally applicable to the treatment of cancer and in reducing the likelihood of development of cancer and/or the metastasis of an existing cancer.
  • the cancer which is treated is metastatic cancer, a recurrent cancer or a drug resistant cancer, especially including a drug resistant cancer.
  • metastatic cancer may be found in virtually all tissues of a cancer patient in late stages of the disease, typically metastatic cancer is found in lymph system/nodes (lymphoma), in bones, in lungs, in bladder tissue, in kidney tissue, liver tissue and in virtually any tissue, including brain (brain cancer/tumor).
  • lymph system/nodes lymph system/nodes
  • the present disclosure is generally applicable and may be used to treat any cancer in any tissue, regardless of etiology.
  • the term “tumor” is used to describe a malignant or benign growth or tumefacent.
  • autoimmune disease refers to a disease or illness that occurs when the body tissues are attacked by its own immune system.
  • the immune system is a complex organization within the body that is designed normally to "seek and destroy" invaders of the body, including infectious agents.
  • MIF levels are often elevated.
  • the present disclosure seeks to inhibit or lower elevated MIF levels in patients with autoimmune disease (as well as inflammatory diseases and conditions and cancer) and by decreasing MIF levels, ameliorate many of the symptoms and secondary effects of these disease states and conditions.
  • autoimmune diseases which often exhibit high expressed levels of MIF including, for example, systemic lupus erythematosus, Sjogren syndrome, Hashimoto thyroiditis, rheumatoid arthritisjuvenile (type 1) diabetes, polymyositis, scleroderma, Addison's disease, vtiligo, pernicious anemia, glomerulonephritis, and pulmonary fibrosis, among numerous others.
  • autoimmune diseases which may be treated by compounds and pharmaceutical compositions according to the present disclosure includes Addison's Disease, Autoimmune polyendodrine syndrome (APS) types 1, 2 and 3, autoimmune pancreatitis (AIP), diabetes mellitus type 1, autoimmune thyroiditis, Ord's thyroiditis, Grave's disease, autoimmune oophoritis, endometriosis, autoimmune orchitis, Sjogren's syndrome, autoimmune enteropathy, coeliac disease, Crohns' disease, microscopic colitis, ulcerative colitis, autophospholipid syndrome (APIS), aplastic anemia, autoimmune hemolytica anemia, autoimmune lymphoproliferative syndrome, autoimmune neutropenia, autoimmune thrombocytopenic purpura, cold agglutinin disease, essential mixed cryoglulinemia, Evans sndrome, pernicious anemia, pure red cell aplasia, thrombocytopenia, adiposis dolorosa, adult-
  • AIP
  • Inflammatory diseases include diseases of neurodegeneration (including, for example, Alzheimer’s disease, Parkinson’s disease, Huntington’s disease; other ataxias), diseases of compromised immune response causing inflammmation (e.g., dysregulation of T cell maturation, B cell and T cell homeostasis, counters damaging inflammation), chronic inflammatory diseases including, for example, inflammatory bowel disease, including Crohn’s disease, rheumatoid arthritis, lupus, multiple sclerosis, chronic obstructive pulmony disease/COPD, pulmonary fibrosis, cystic fibrosis, Sjogren’s disease; hyperglycemic disorders, diabetes (I and II), affecting lipid metabolism islet function and/or structure, pancreatic P-cell death and related hyperglycemic disorders, including severe insulin resistance, hyperinsulinemia, insulin-resistant diabetes (e.g.
  • dyslipidemia e.g. hyperlipidemia as expressed by obese subjects, elevated low-density lipoprotein (LDL), depressed high- density lipoprotein (HDL), elevated triglycerides and metabolic syndrome, liver disease, renal disease (apoptosis in plaques, glomerular disease), cardiovascular disease (especially including infarction, ischemia, stroke, pressure overload and complications during reperfusion), muscle degeneration and atrophy, low grade inflammation, gout, silicosis, atherosclerosis and associated conditions such as cardiac and neurological (both central and peripheral) manifestations including stroke, age-associated dementia and sporadic form of Alzheimer's disease, and psychiatric conditions including depression), stroke and spinal cord injury, arteriosclerosis, among others.
  • dyslipidemia e.g. hyperlipidemia as expressed by obese subjects, elevated low-density lipoprotein (LDL), depressed high- density lipoprotein (HDL), elevated triglycerides and metabolic syndrome, liver disease, renal disease (apoptosis in plaques, glomerular
  • linker refers to a chemical entity including a complex linker connecting a macrophage migration inhibitory factor binding moiety (MIFBM) to the asialoglycoprotein receptor binding moiety (ASGPRBM), optionally through at least one (such as one or two) connector moiety [CON] through covalent bonds in compounds according to the present disclosure.
  • MIFBM macrophage migration inhibitory factor binding moiety
  • ASGPRBM asialoglycoprotein receptor binding moiety
  • the linker between the two active portions of the molecule ranges from about 5 A to about 50A or more in length, about 6A to about 45 A in length, about 7 A to about 40 A in length, about 8 A to about 35 A in length, about 9 A to about 30A in length, about 10A to about 25 A in length, about 7 A to about 20 A in length, about 5 A to about 16A in length, about 5 A to about 15A in length, about 6 A to about 14A in length, about 10A to about 20 A in length, about 11 A to about 25 A in length, etc.
  • Linkers which are based upon ethylene glycol units and are between 2 and 15 glycol units, 1 and 8 glycol units, 1, 2, 3, 4, 5, and 6 glycol units in length may be used, although the length of certain linkers may be far greater.
  • the MIFBM group and the ASGPRBM group may be situated to advantageously take advantage of the biological activity of compounds according to the present disclosure which bind to asialoglycoprotein receptors on hepatocytes resulting in the selective and targeted degradation of MIF circulating proteins within the lysosomal degradation mechanism or other degradation mechanism of the hepatocytes.
  • linker component The selection of a linker component is based on its documented properties of biocompatibility, solubility in aqueous and organic media, and low immunogenicity/antigenicity. Although numerous linkers may be used as otherwise described herein, a linker based upon polyethyleneglycol (PEG) linkages, polypropylene glycol linkages, or poly ethyl eneglycol-co-polypropylene oligomers (up to about 100 units, about 1 to 100, about 1 to 75, about 1 to 60, about 1 to 50, about 1 to 35, about 1 to 25, about 1 to 20, about 1 to 15, 2 to 10, about 4 to 12, about 1 to 8, 1 to 3, 1 to 4, 2 to 6, 1 to 5, etc.) may be favored as a linker because of the chemical and biological characteristics of these molecules.
  • PEG polyethyleneglycol
  • polypropylene glycol linkages or poly ethyl eneglycol-co-polypropylene oligomers (up to about 100 units, about
  • Polyethylene (PEG) linkages of between 2 and 15 ethylene glycol units are sometimes used.
  • one or more additional groups e.g., methylene groups, amide groups, keto groups, amine groups, etc.
  • Alternative linkers may include, for example, polyamino acid linkers of up to 100 amino acids (of any type, such as D- or L- amino acids, such as naturally occurring L-amino acids) in length (about 1 to 75, about 1 to 60, about 1 to 50, about 1 to 45, about 1 to 35, about 1 to 25, about 1 to 20, about 1 to 15, 2 to 10, about 4 to 12, about 5 to 10, about 4 to 6, about 1 to 8, about 1 to 6 , about 1 to 5, about 1 to 4, about 1 to 3, etc. in length), optionally including one or more connecting groups (such as 1 or 2 connecting groups at one or both ends of the polyamino acid linker).
  • polyamino acid linkers of up to 100 amino acids (of any type, such as D- or L- amino acids, such as naturally occurring L-amino acids) in length (about 1 to 75, about 1 to 60, about 1 to 50, about 1 to 45, about 1 to 35, about 1 to 25, about 1 to 20, about 1 to 15, 2 to 10, about 4 to 12, about 5 to 10, about
  • Exemplary linkers include those according to the chemical structures: or a polypropylene glycol or polypropylene-co-polyethylene glycol linker having between 1 and 100 alkylene glycol units, such as about 1 to 75, about 1 to 60, about 1 to 50, about 1 to 45, about 1 to 35, about 1 to 25, about 1 to 20, about 1 to 15, 2 to 10, about 4 to 12, about 5 to 10, about 4 to 6, about 1 to 8, about 1 to 6, about 1 to 5, about 1 to 4, about 1 to 3; where Ra is H, C1-C3 alkyl or alkanol or forms a cyclic ring with R 3 (proline) and R 3 is a side chain derived from a D- or L amino acid (such as a naturally occurring L-amino acid), such as selected from the group consisting of alanine (methyl), arginine (propyleneguanidine), asparagine (methylenecarboxyamide), aspartic acid (ethanoic acid), cysteine (thiol, reduced or oxidized
  • linkers include a polyethylene glycol linker containing from 1 to 1 to 100, such as 1 to 75, 1 to 60, 1 to 55, 1 to 50, 1 to 45, 1 to 40, 2 to 35, 3 to 30, 1 to 15, 1 to 10, 1 to 8, 1 to 6, 1, 2, 3, 4 or 5 ethylene glycol units, to which is bonded a lysine group or other amino acid moiety at one or both ends of the linker (which can consist of between 1 and 10 amino acids which can bind the MIFBM and/or the ASGPRBM group.
  • Still other linkers comprise amino acid residues (D or L) which are bonded to MIFBM and/or ASGPRBM moieties as otherwise described herein.
  • the amino acid has anywhere from 1-15 methylene groups separating the amino group from the acid (acyl) group in providing a linker to the MIFBM and/or the ASGPRBM group, wherein the linker contains from 1 to 100, 1 to 75, 1 to 60, 1 to 55, 1 to 50, 1 to 45, 1 to 40, 2 to 35, 3 to 30, 1 to 15, 1 to 10, 1 to 8, 1 to 6, 1, 2, 3, 4 or 5 amino acid groups linked together through peptide linkages to form the linker.
  • This linker is represented by the chemical structure: where Ram is H or a C1-C3 alkyl optionally substituted with one or two hydroxyl groups; na is an integer ranging from 1 to 15, such as 1-12, 1-10, 1-8, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11; m is an integer ranging from 1 to 100, such as 1 to 75, 1 to 60, 1 to 55, 1 to 50, 1 to 45, 1 to 40, 2 to 35, 3 to 30, 1 to 15, 1 to 12, 1 to 10, 1 to 8, 1 to 6, 1, 2, 3, 4 or 51 to 50, 1 to 45, 1 to 40, 2 to 35, 3 to 30, 1 to 15, 1 to 10, 1 to 8, 1 to 6, 1, 2, 3, 4 or 5.
  • Z and Z’ are each independently a bond, -(CH2)i-O, -(CH2)i-S, -(CH2)i-N-R, wherein said -(CH2)i group, if present in Z or Z’, is bonded to a connector (CON), MIFBM or ASGPRBM; each R is H, or a C1-C3 alkyl or alkanol group; each R 2 is independently H or a C1-C3 alkyl group; each Y is independently a bond, O, S or N-R; each i is independently an integer ranging from 0 to 100, such as 0 to 75, 1 to 60, 1 to 55, 1 to 50, 1 to 45, 1 to 40, 2 to 35, 3 to 30, 1 to 15, 1 to 10, 1 to 8, 1 to 6, 0, 1, 2, 3, 4 or 5;
  • D is a bond, with the proviso that Z, Z’ and D are not each simultaneously bonds; j is an integer ranging from 1 to 100, such as 1 to 75, 1 to 60, 1 to 55, 1 to 50, 1 to 45,
  • m’ is an integer ranging from 1 to 100, such as 1 to 75, 1 to 60, 1 to 55, 1 to 50, 1 to
  • n is an integer ranging from 1 to 100, such as 1 to 75, 1 to 60, 1 to 55, 1 to 50, 1 to 45, 1 to 40, 2 to 35, 3 to 30, 1 to 15, 1 to 10, 1 to 8, 1 to 6, 1, 2, 3, 4 or 5;
  • X 1 is O, S or N-R
  • R is H, or a C1-C3 alkyl or alkanol group, or a pharmaceutical salt thereof.
  • linkers which are included herein include linkers according to the chemical structure: where each n and n’ is independently an integer ranging from 1 to 25, such as 1 to 15, 1 to 12, 2 to 11, 2 to 10, 2 to 8, 2 to 6, 2 to 5, 2 to 4 and 2 to 3 or 1, 2, 3, 4, 5, 6, 7, or 8; and each n” is independently an integer ranging from 0 to 8, such as 1 to 7, or 1, 2, 3, 4, 5 or 6.
  • Linkers also can comprise two or more linker segments (based upon the linkers described above) which are attached directly to each other or through [CON] groups forming a complex linker.
  • Certain linkers which include a [CON] group (especially a diamide [CON] group as otherwise described herein) connecting a first and second (PEG) linker group include the following structures: where each n and n’ is independently an integer ranging from 1 to 25, such as 1 to 15, 1 to 12, 2 to 11, 2 to 10, 2 to 8, 2 to 6, 2 to 5, 2 to 4 and 2 to 3 or 1, 2, 3, 4, 5, 6, 7, or 8; and each n” is independently an integer ranging from 0 to 8, such as 1 to 7, or 1, 2, 3, 4, 5 or 6. Noted is that each of these linkers may also contain alkylene groups containing from 1 to 4 methylene groups at the distal ends of each linker group in order to facilitate connection of the linker group.
  • linkers which include a connector group [CON] include groups which are represented by the chemical formula PEG-[CON]-PEG wherein each PEG is independently a polyethylene glycol group containing from 1-12 ethylene glycol residues and [CON] is a connector group as otherwise set forth herein, such as a triazole group
  • connector symbolized in the generic formulas by “CON” or [CON], is used to describe a chemical moiety which is optionally included in bifunctional compounds according to the present disclosure which forms from the reaction product of an activated linker with a MIFBM moiety (which also is sometimes activated for covalently bonding the linker with the moiety) or a ASGPRBM group with an activated linker.
  • the connector group is often the resulting moiety which forms from the facile condensation of two or more separate chemical fragments which contain reactive groups which can provide connector groups as otherwise described to produce bifunctional or multifunctional compounds according to the present disclosure.
  • a connector may be distinguishable from a linker in that the connector is the result of a specific chemistry which is used to provide bifunctional compounds according to the present disclosure wherein the reaction product of these groups results in an identifiable connector group or part of a connector group which is distinguishable from the linker group, although in certain instances, the connector group is incorporated into and integral with the linker group as otherwise described herein. It is noted also that a connector group may be linked to a number of linkers to provide multifunctionality (i.e., more than one CPBM moiety and/or more than one ASGPRBM/FCRNBM moiety) within the same molecule.
  • a connector may be connected to MIFBM, ASGPRBM or a linker at positions which are represented as being linked to another group using the symbol
  • any of an ASGPRBM, a linker or a MIFBM may be bonded to such a group.
  • the linker may be at one or more positions of a moiety.
  • R CON1 and R C0N2 are each independently H, methyl or a bond (for attachment to another moiety); or a diamide group according to the structure: where X 2 is CH 2 , O, S, NR 4 , C(O), S(O), S(O) 2 , -S(O) 2 O, -OS(O) 2 , or OS(O) 2 O;
  • X 3 is O, S, NR 4 ;
  • R 4 is H, a C1-C3 alkyl or alkanol group, or a -C(O)(Ci-C3) group;
  • Ri is H or a C1-C3 alkyl group; and n” is independently an integer ranging from 0 to 8, such as 1 to 7, or 1, 2, 3, 4, 5 or 6; or the connector group [CON] is a group according to the chemical structure: where R 1C0N , R 2CON and R 3C0N are each independently H, -(CH 2 )MCI, -
  • each MCI is independently an integer ranging from 1 to 4, such as 1 or 2; each MCI a is independently an integer ranging from 0 to 4, such as 0, 1 or 2; and R 4 is H, a C1-C3 alkyl or alkanol group, or a -C(O)(Ci-C3) group.
  • the triazole group is an exemplary connector group.
  • An additional exemplary connector group is which is linked to at least one MIFBM and/or at least one ASPRGBM (such as 3 ASPRGBM moieties). This connector group may be used to form GN3.
  • each connector may be extended with one or more methylene groups to facilitate connection to a linker group, another CON group, a MIFBM group or a ASGPRBM. It is noted that in certain instances, within context the diamide group may also function independently as a linker group. Additional Galactose- and Talose-based ASGPR Binding Moieties
  • the present disclosure is directed to compounds which are useful for removing circulating proteins which are associated with a disease state or condition in a patient or subject according to the general chemical structure of Formula II:
  • Extracellular Protein Targeting Ligand as used herein is interchangeably used with the term CPBM (cellular protein binding moiety).
  • ASGPR Ligand as used herein is interchangeably used with an asialoglycoprotein receptor (ASGPR) binding moiety as defined herein.
  • each [CON] is an optional connector chemical moiety which, when present, connects directly to [CPBM] or to [CRBM] or connects the [LINKER- 2] to [CPBM] or to [CRBM],
  • [LINKER-2] is a chemical moiety having a valency from 1 to 15 which covalently attaches to one or more [CRBM] and/or [CPBM] group, optionally through a [CON], including a [MULTICON] group, wherein said [LINKER-2] optionally itself contains one or more [CON] or [MULTICON] group(s); k’ is an integer from 1 to 15; j’ is an integer from 1 to 15; h and h’ are each independently an integer from 0 to 15;
  • IL is an integer from 0 to 15; with the proviso that at least one of h, h’ and 11 is at least 1, or a pharmaceutically acceptable salt, stereoisomer, solvate or polymorph thereof.
  • a [MULTICON] group can connect one or more of a [CRBM] or [CPBM] to one or more of a [LINKER-2],
  • [LINKER-2] has a valency of 1 to 10.
  • [LINKER-2] has a valency of 1 to 5.
  • [LINKER-2] has a valency of 1, 2 or 3.
  • the [LINKER-2] includes one or more of Linker ⁇ Linker®, Linker 13 , Linker 0 , and/or combinations thereof as defined herein.
  • xx is independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, and 25.
  • yy is independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, and 25.
  • zz is independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, and 25.
  • X 1 is 1 to 5 contiguous atoms independently selected from O, S, N(R b ), and C(R 4 )(R 4 ), wherein if X 1 is 1 atom then X 1 is O, S, N(R 6 ), or C(R 4 )(R 4 ), if X 1 is 2 atoms then no more than 1 atom of X 1 is O, S, or N(R 6 ), if X 1 is 3, 4, or 5 atoms then no more than 2 atoms of X 1 are O, S, or N(R 6 );
  • R 3 at each occurrence is independently selected from hydrogen, alkyl, heteroalkyl, haloalkyl (including -CF3, -CHF2, -CH2F, -CH2CF3, -CH2CH2F, and -CF2CF3), arylalkyl, heteroarylalkyl, alkenyl, alkynyl, and, heteroaryl, heterocycle, -OR 8 , and -NR 8 R 9 ;
  • R 4 is independently selected at each occurrence from hydrogen, heteroalkyl, alkyl, haloalkyl, arylalkyl, heteroarylalkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocycle, -OR 6 , - NR 6 R 7 ,
  • R 6 and R 7 are independently selected at each occurrence from hydrogen, heteroalkyl, alkyl, arylalkyl, heteroaryl alkyl, alkenyl, alkynyl, and, haloalkyl, heteroaryl, heterocycle, - alkyl-OR 8 , -alkyl-NR 8 R 9 , C(O)R 3 , S(O)R 3 , C(S)R 3 , and S(O) 2 R 3 ;
  • R 8 and R 9 are independently selected at each occurrence from hydrogen, heteroalkyl, alkyl, arylalkyl, heteroarylalkyl, alkenyl, alkynyl, aryl, heteroaryl, and heterocycle.
  • the compound of Formula II is selected from:
  • the compound of Formula II has one of the following structures:
  • the ASGPR ligand is linked at either the C 1 or C 5 (R 1 or R 5 ) position to form a degrading compound. In various embodiments, the ASGPR ligand is linked at C 6 position to form a degrading compound.
  • ASGPR binding compounds of Formula II include: or the bi- or tri- substituted versions thereof or pharmaceutically acceptable salts thereof, where the bi- or tri- substitution refers to the number additional galactose derivatives attached to a linker moiety.
  • an ASGPR ligand is drawn for use in a degrader the ASGPR ligand is typically linked through to the Extracellular Protein Targeting Ligand in the C 5 position (e.g., which can refer to the adjacent C 6 carbon hydroxyl or other functional moiety that can be used for linking purposes).
  • the linker and Extracellular Protein Targeting Ligand is connected through the C 1 position, then that carbon is appropriately functionalized for linking, for example with a hydroxyl, amino, allyl, alkyne or hydroxyl-allyl group.
  • the ASGPR ligand is not linked in the C 3 or C 4 position, because these positions chelate with the calcium for ASGPR binding in the liver.
  • an ASGPR ligand useful for incorporation into a compound of Formula II is selected from:
  • the compound of Formula II is selected from:
  • the compound of Formula II is selected from:
  • the compound of Formula II is an Extracellular Protein degrading compound in which the ASGPR ligand is a ligand as described herein
  • the ASGPR ligand in the compound of Formula II, is linked at either the Cl or C5 (R 1 or R 5 ) position to form a degrading compound. In certain embodiments, in the compound of Formula II, the ASGPR ligand is linked at C6. In various embodiments, when the ASGPR ligand is then non- limiting examples of ASGPR binding compounds of Formula II include:
  • the compound of Formula II is selected from:
  • R 2 is selected from -NR 6 COR 3 , -NR 6 -(5-membered heteroaryl), and-NR 6 -(6-membered heteroaryl), each of which R 2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
  • the compound of Formula II is selected from:
  • R 2 is selected from -NR 6 COR 3 , -NR 6 -(5-membered heteroaryl), and-NR 6 -(6-membered heteroaryl), each of which R 2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
  • the compound of Formula II is selected from:
  • R 2 is selected from -NR 6 COR 3 , -NR 6 -(5-membered heteroaryl), and-NR 6 -(6-membered heteroaryl), each of which R 2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
  • the compound of Formula II is selected from:
  • R 2 is selected from -NR 6 COR 3 , -NR 6 -(5-membered heteroaryl), and-NR 6 -(6-membered heteroaryl), each of which R 2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
  • the compound of Formula II is selected from:
  • R 2 is selected from -NR 6 COR 3 , -NR 6 -(5-membered heteroaryl), and-NR 6 -(6-membered heteroaryl), each of which R 2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
  • the compound of Formula II is selected from:
  • R 2 is selected from -NR 6 COR 3 , -NR 6 -(5-membered heteroaryl), and-NR 6 -(6-membered heteroaryl), each of which R 2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
  • the compound of Formula II is selected from:
  • R 2 is selected from -NR 6 COR 3 , -NR 6 -(5-membered heteroaryl), and-NR 6 -(6-membered heteroaryl), each of which R 2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
  • the compound of Formula II is selected from:
  • R 2 is selected from -NR 6 COR 10 , -NR 6 -(5-membered heteroaryl), and-NR 6 -(6-membered heteroaryl), each of which R 2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
  • the compound of Formula II is selected from:
  • R 2 is selected from -NR b COR 10 , -NR 6 -(5-membered heteroaryl), and-NR 6 -(6-membered heteroaryl), each of which R 2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
  • the compound of Formula II is selected from:
  • R 2 is selected from -NR 6 COR 10 , -NR 6 -(5-membered heteroaryl), and-NR 6 -(6-membered heteroaryl), each of which R 2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
  • the compound of Formula II is selected from:
  • R 2 is selected from -NR 6 COR 10 , -NR 6 -(5-membered heteroaryl), and-NR 6 -(6-membered heteroaryl), each of which R 2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
  • the compound of Formula II is selected from:
  • R 2 is selected from -NR 6 COR 10 , -NR 6 -(5-membered heteroaryl), and-NR 6 -(6-membered heteroaryl), each of which R 2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
  • the compound of Formula II is selected from:
  • R 2 is selected from -NR 6 COR 10 , -NR 6 -(5-membered heteroaryl), and-NR 6 -(6-membered heteroaryl), each of which R 2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
  • the compound of Formula II is selected from:
  • R 2 is selected from -NR 6 COR 10 , -NR 6 -(5-membered heteroaryl), and-NR 6 -(6-membered heteroaryl), each of which R 2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
  • the compound of Formula II is selected from:
  • an ASGPR ligand useful for incorporation into a compound of Formula II is selected from:
  • R 1 is hydrogen
  • R 1 is
  • R 1 is
  • R 1 is
  • R 1 is
  • R 1 is
  • R 1 is
  • R 1 is Co-Cealkyl-cyano optionally substituted with 1, 2, 3, or 4 substituents.
  • R 1 is alkyl optionally substituted with 1, 2, 3, or 4 substituents.
  • R 1 is alkenyl optionally substituted with 1, 2, 3, or 4 substituents. In certain embodiments, in the compound of Formula II, R 1 is alkynyl optionally substituted with 1, 2, 3, or 4 substituents. In certain embodiments, in the compound of Formula II, R 1 is haloalkyl optionally substituted with 1, 2, 3, or 4 substituents. In certain embodiments, in the compound of Formula II, R 1 is F.
  • R 1 is Cl
  • R 1 is Br
  • R 1 is aryl optionally substituted with 1, 2, 3, or 4 substituents.
  • R 1 is arylalkyl optionally substituted with 1, 2, 3, or 4 substituents.
  • R 1 is heteroaryl optionally substituted with 1, 2, 3, or 4 substituents.
  • R 1 is heteroaryl alkyl optionally substituted with 1, 2, 3, or 4 substituents.
  • R 1 is heterocycle optionally substituted with 1, 2, 3, or 4 substituents.
  • R 1 is heterocycloalkyl optionally substituted with 1, 2, 3, or 4 substituents. In certain embodiments, in the compound of Formula II, R 1 is haloalkoxy optionally substituted with 1, 2, 3, or 4 substituents.
  • R 1 is -O-alkenyl, -O-alkynyl, C0-C6alkyl-OR 6 , C0-C6alkyl-SR 6 , C0-C6alkyl-NR 6 R 7 , C0-C6alkyl-C(O)R 3 , C0-C6alkyl-S(O)R 3 , C0-C6alkyl-C(S)R 3 , C0-C6alkyl-S(O)2R 3 , C0-C6alkyl-N(R 8 )-C(O)R 3 , C0-C6alkyl-N(R 8 )- S(O)R 3 , C 0 -C 6 alkyl-N(R 8 )-C(S)R 3 , C 0 -C 6 alkyl-N(R 8 )-S(O) 2 R 3 C 0 -C 6 alkyl-O-C(O)R 3 ,
  • R 2 is aryl optionally substituted with 1, 2, 3, or 4 substituents. In certain embodiments, in the compound of Formula II, R 2 is heterocycle optionally substituted with 1, 2, 3, or 4 substituents. In certain embodiments, in the compound of Formula II, R 2 is heteroaryl containing 1 or 2 heteroatoms independently selected from N, O, and S optionally substituted with 1, 2, 3, or 4 substituents. In certain embodiments, in the compound of Formula II, R 2 is selected from In certain embodiments, in the compound of Formula II, R 2 is heterocycle optionally substituted with 1, 2, 3, or 4 substituents.
  • R 2 is -NR 8 -S(O) 2 -R 10 optionally substituted with 1, 2, 3, or 4 substituents.
  • R 2 is -NR 8 -C(NR 6 )-R 3 optionally substituted with 1, 2, 3, or 4 substituents.
  • R 2 is hydrogen
  • R 2 is R 10 .
  • R 2 is alkyl-C(O)-R 3 .
  • R 2 is -C(O)-R 3 .
  • R 2 is alkyl
  • R 2 is haloalkyl
  • R 2 is -OC(O)R 3 .
  • R 2 is -NR 8 -C(O)R 10 .
  • R 2 is alkenyl optionally substituted with 1, 2, 3, or 4 substituents.
  • R 2 is allyl optionally substituted with 1, 2, 3, or 4 substituents.
  • R 2 is alkynyl optionally substituted with 1, 2, 3, or 4 substituents.
  • R 2 is -NR 6 -alkenyl optionally substituted with 1, 2, 3, or 4 substituents.
  • R 2 is -O-alkenyl optionally substituted with 1, 2, 3, or 4 substituents.
  • R 2 is -NR 6 -alkynyl optionally substituted with 1, 2, 3, or 4 substituents.
  • R 2 is -NR 6 -heteroaryl optionally substituted with 1, 2, 3, or 4 substituents.
  • R 2 is -NR 6 -aryl optionally substituted with 1, 2, 3, or 4 substituents.
  • R 2 is -O-heteroaryl optionally substituted with 1, 2, 3, or 4 substituents.
  • R 2 is -O-aryl optionally substituted with 1, 2, 3, or 4 substituents.
  • R 2 is -O-alkynyl optionally substituted with 1, 2, 3, or 4 substituents.
  • R 2 is selected from and
  • R 2 is selected from
  • R 2 is selected from
  • R is an optional substituent as defined herein.
  • R 2 is selected from
  • R 2A is selected from an optional substituent as defined herein.
  • R 2A is selected from
  • R 2 is selected from ,
  • R 2 is selected from
  • R 2 is selected from In certain embodiments, in the compound of Formula II, R 2 is selected from
  • R 2 is selected from
  • R 2 is selected from
  • R 2 is selected from
  • R 2 is selected from
  • R 2 is selected from In certain embodiments, in the compound of Formula II, R 2 is selected from
  • R 2 is selected from
  • R 2 is selected from
  • R 2 is selected from
  • R 2 is selected from
  • R 2 is selected from
  • R 2 is selected from
  • R 2 is selected from
  • R 2 is selected from
  • R 2 is selected from
  • R 2 is selected from
  • R 2 is selected from
  • R 2 is selected from
  • R 2 is selected from
  • R 2 is selected from
  • R 2 is selected from In certain embodiments, in the compound of Formula II, R 2 is selected from
  • R 2 is selected from In certain embodiments, in the compound of Formula II, R 2 is selected from
  • R 2 is selected from
  • R 2 or R 2A is selected from
  • R 2 is selected from
  • R 2 is selected from
  • R 2 is selected from
  • R 2 is selected from In certain embodiments, in the compound of Formula II, R 2 is selected from In certain embodiments, in the compound of Formula II, R 2 is selected from In certain embodiments, in the compound of Formula II, R 2 is a spirocyclic heterocycle, for example, and without limitation,
  • R 2 is a silicon containing heterocycle, for example, and without limitation, in certain embodiments, in the compound of Formula II, R 2 is substituted with SFs, for example, and without limitation,
  • R 2 is substituted with a sulfoxime, for example, and without limitation, in certain embodiments, in the compound of Formula II, R 10 is selected from bicyclic heterocycle.
  • R 10 is selected from spirocyclic heterocycle.
  • R 10 is selected from -NR 6 - heterocycle.
  • R 10 is selected from
  • R 10 is selected from
  • R is selected from
  • R 10 is selected from
  • Cycle is selected from
  • R 30 is selected from:
  • R 200 is
  • R is
  • R 200 is
  • R 200 is
  • R is
  • R 200 is
  • R 200 is
  • R 200 is
  • R 200 is
  • R 200 is :
  • R 200 is
  • R 200 is
  • Linker A and Linker® are independently selected from: wherein:
  • R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 , and R 20 are independently at each occurrence selected from the group consisting of a bond, alkyl, -C(O)-, -C(O)O-, -OC(O)-, -SO2-, -S(O)-, -C(S)-, -C(O)NR 6 -, -NR 6 C(O)-, -O-, -S-, -NR 6 -, -C(R 21 R 21 )-, -P(O)(R 3 )O-, -P(O)(R 3 )-, a divalent residue of a natural or unnatural amino acid, alkenyl, alkynyl, haloalkyl, alkoxy, and, heterocycle, heteroaryl, -CH 2 CH2-[O-(CH 2 )2]n-O-, CH 2 CH2-
  • n is independently selected at each instance from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;
  • R 21 is independently at each occurrence selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, F, Cl, Br, I, hydroxyl, alkoxy, azide, amino, cyano, - NR 6 R 7 , -NR 8 SO2R 3 , -NR 8 S(O)R 3 , haloalkyl, heteroalkyl, and, heteroaryl, and heterocycle; and the remaining variables are as defined herein.
  • Linker A is bond and Linker 8 is
  • Linker 8 is bond and Linked is
  • a divalent residue of an amino acid is selected from
  • a divalent residue of a dicarboxylic acid is generated from a nucleophilic addition reaction:
  • Non-limiting embodiments of a divalent residue of a dicarboxylic acid generated from a nucleophilic addition reaction include:
  • a divalent residue of a dicarboxylic acid is generated from a condensation reaction:
  • Non-limiting embodiments of a divalent residue of a saturated dicarboxylic acid include: Non-limiting embodiments of a divalent residue of a saturated monocarboxylic acid is selected from butyric acid (-OC(O)(CH2)2CH2-), caproic acid (-OC(O)(CH2)4CH2-), caprylic acid (-OC(O)(CH2)SCH2-), capric acid (-OC(O)(CH2)sCH2-), lauric acid (- OC(0)(CH2)IOCH2-), myristic acid (-OC(O)(CH2)i2CH2-), pentadecanoic acid (- OC(O)(CH 2 )i3CH 2 -), palmitic acid (-OC(O)(CH2)i4CH2-), stearic acid (-OC(O)(CH2)ieCH2-), behenic acid (-OC(0)(CH2)2oCH2-), and lignoceric acid (-OC(O)(CH2)22CH2-);
  • Non-limiting embodiments of a divalent residue of a fatty acid include residues selected from linoleic acid, palmitoleic acid, vaccenic acid, paullinic acid, oleic acid, elaidic acid, gondoic acid, gadoleic acid, nervonic acid, myristoleic acid, and erucic acid:
  • Non-limiting embodiments of a divalent residue of a fatty acid is selected from linoleic acid (-C(O)(CH2)?(CH)2CH2(CH)2(CH2)4CH2-), docosahexaenoic acid (-C(O)(CH2)2(CHCHCH2)eCH2-), eicosapentaenoic acid (- C(O)(CH2)3(CHCHCH 2 ) 5 CH2-), alpha-linolenic acid (-C(O)(CH 2 )7(CHCHCH 2 )3CH2-) stearidonic acid
  • Linker 13 is selected from: wherein:
  • R 22 is independently at each occurrence selected from the group consisting of alkyl, - C(O)N-, -NC(O)-, -N-, -C(R 21 )-, -P(O)O-, -P(O)-, -P(O)(NR 6 R 7 )N-, alkenyl, haloalkyl, aryl, heterocycle, and heteroaryl, each of which is optionally substituted with 1, 2, 3, or 4 substituents independently selected from R 21 ; and the remaining variables are as defined herein.
  • Linker 0 is selected from: wherein:
  • R 32 is independently at each occurrence selected from the group consisting of alkyl, N + X-, -C-, alkenyl, haloalkyl, aryl, heterocycle, and heteroaryl, each of which is optionally substituted with 1, 2, 3, or 4 substituents independently selected from R 21 ;
  • X- is an anionic group, for example Br- or Cl‘ ; and all other variables are as defined herein.
  • Linked is selected from: wherein each heteroaryl, heterocycle, cycloalkyl, and aryl can optionally be substituted with 1, 2, 3, or 4 of any combination of halogen, alkyl, haloalkyl, and, heteroaryl, heterocycle, or cycloalkyl, as allowed by valence.
  • Linked is selected from:
  • Linker 8 is selected from:
  • Linker 8 is selected from:
  • Linker 8 in the compound of Formula II, is selected from:
  • tt is independently selected from 1, 2, or 3 and ss is 3 minus tt (3-tt).
  • Linker 8 in the compound of Formula II, Linker 8 , Linker 13 , or Linker 0 ted from: wherein tt and ss are as defined herein.
  • Linker 8 in the compound of Formula II, Linker 8 , Linker 13 , or Linker 0 ted from:
  • each heteroaryl, heterocycle, cycloalkyl, and aryl can optionally be substituted with 1, 2, 3, or 4 of any combination of halogen, alkyl, haloalkyl, aryl, heteroaryl, heterocycle, or cycloalkyl, as allowed by valence; and tt and ss are as defined herein.
  • Linker 8 in the compound of Formula II, Linker 8 , Linker 13 , or Linker 0 is selected from:
  • each heteroaryl, heterocycle, cycloalkyl, and aryl can optionally be substituted with 1, 23, or 4 of any combination of halogen, alkyl, haloalkyl, and, heteroaryl, heterocycle, or cycloalkyl, as allowed by valence: and tt and ss are as defined herein.
  • Linker B , Linker C , or Linker D is selected from:
  • Linker A is selected from:
  • Linked is selected from: In certain embodiments, in the compound of Formula II, Linked is selected from:
  • Linker A is selected from: In certain embodiments, in the compound of Formula II, Linker B is selected from: In certain embodiments, in the compound of Formula II, Linker B is selected from:
  • Linker B is selected from:
  • Linker B is selected from:
  • Linker C is selected from: In certain embodiments, in the compound of Formula II, Linker C is selected from:
  • Linker C is selected from:
  • Linker C is selected from: In certain embodiments, in the compound of Formula II, Linker C is selected from:
  • Linker C is selected from: In certain embodiments, in the compound of Formula II, Linker C is selected from:
  • Linker C is selected from:
  • Linker D is selected from:
  • LinkerD is selected from: In certain embodiments, in the compound of Formula II, Linker D is selected from: In certain embodiments, in the compound of Formula II, Linker D is selected from:
  • Linker D is selected from: In certain embodiments, in the compound of Formula II, Linker D is selected from:
  • the Linker A is selected from
  • the Linker A is selected from
  • the Linker A is selected from
  • the Linker A is selected from wherein each is optionally substituted with 1, 2, 3, or 4 substituents substituent selected from R 21 .
  • Linked is selected from:
  • the Linker A is selected from
  • the Linker A is selected from
  • the Linker A is selected from
  • the Linker A is selected from
  • the Linker A is selected from
  • the Linker A is selected from In certain embodiments, in the compound of Formula II, the Linker A is selected from
  • the Linker A is selected from
  • the Linker A is selected from
  • the Linker A is selected from
  • the Linker A is selected from
  • the Linker A is selected from
  • the Linker A is selected from In certain embodiments, in the compound of Formula II, the Linker A is selected from In certain embodiments, in the compound of Formula II, the Linker B is selected from In certain embodiments, in the compound of Formula II, the Linker B is selected from In certain embodiments, in the compound of Formula II, the Linker B is selected from In certain embodiments, in the compound of Formula II, the Linker B is selected from wherein each is optionally substituted with 1, 2, 3, or 4 substituents substituent selected from R 21 .
  • Linker B is selected from: In certain embodiments, in the compound of Formula II, the Linker B is selected from: In certain embodiments, in the compound of Formula II, the Linker B is selected from: In certain embodiments, in the compound of Formula II, the Linker B is selected from: In certain embodiments, in the compound of Formula II, the Linker B is selected from: In certain embodiments, in the compound of Formula II, the Linker B is selected from: In certain embodiments, in the compound of Formula II, the Linker B is selected from: In certain embodiments, in the compound of Formula II, the Linker B is selected from:
  • the Linker B is selected from: In certain embodiments, in the compound of Formula II, the Linker B is selected from:
  • Linker B -Linker A is selected from: In certain embodiments, in the compound of Formula II, Linker B -Linker A is selected from: In certain embodiments, in the compound of Formula II, the Linker C is selected from: In certain embodiments, in the compound of Formula II, the Linker C is selected from:
  • the Linker C is selected from: In certain embodiments, in the compound of Formula II, the Linker C is selected from: In certain embodiments, in the compound of Formula II, the Linker C is selected from: In certain embodiments, in the compound of Formula II, the Linker C is selected from: In certain embodiments, in the compound of Formula II, the Linker C is selected from: In certain embodiments, in the compound of Formula II, the Linker C is selected from:
  • the Linker C is selected from: wherein each is optionally substituted with 1, 2, 3, or 4 substituents substituent selected from R 21 . In certain embodiments, in the compound of Formula II, the Linker C is selected from: In certain embodiments, in the compound of Formula II, the Linker C is selected from: In certain embodiments, in the compound of Formula II, the Linker C is selected from: In certain embodiments, in the compound of Formula II, the Linker C is selected from: In certain embodiments, in the compound of Formula II, the Linker C is selected from: In certain embodiments, in the compound of Formula II, the Linker C is selected from: In certain embodiments, in the compound of Formula II, the Linker C is selected from: In certain embodiments, in the compound of Formula II, the Linker C is selected from:
  • the Linker C is selected from: In certain embodiments, in the compound of Formula II, Linker C -(Linker A )2 is selected from: In certain embodiments, in the compound of Formula II, Linker C -(Linker A )2 is selected from: In certain embodiments, in the compound of Formula II, Linker C -(Linker A )2 is selected from: In certain embodiments, in the compound of Formula II, Linker C -(Linker A ) 2 is selected from: In certain embodiments, in the compound of Formula II, Linker C -(Linker A ) 2 is selected from:
  • Linker D is selected from:
  • Linker D is selected from: wherein each is optionally substituted with 1, 2, 3, or 4 substituents are selected from R 21 .
  • Linker B -(Linker A ) is selected from
  • Linker c -(Linker A ) is selected from
  • Linker°-(Linker A ) is selected from
  • R 4 is independently selected at each occurrence from hydrogen, heteroalkyl, alkyl, haloalkyl, arylalkyl, heteroarylalkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocycle, -OR 6 , -NR 6 R 7 , C(O)R 3 , S(O)R 3 , C(S)R 3 , and S(O)2R 3 .
  • R 5 is independently selected from hydrogen, heteroalkyl, , C 0 -C 6 alkyl-cyano, alkyl, alkenyl, alkynyl, haloalkyl, F, Cl, Br, I, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocycle, heterocycloalkyl, haloalkoxy, -O-alkenyl, -O-alkynyl, C 0 -C 6 alkyl- OR 6 , C 0 -C 6 alkyl-SR 6 , C 0 - C 6 alkyl-NR 6 R 7 , C 0 -C 6 alkyl-C(O)R 3 , C 0 -C 6 alkyl-S(O)R 3 , C 0 -C 6 alkyl- C(S)R 3 , C 0 -C 6 alkyl- S(O)2R 3 , C0-
  • R 6 and R 7 are independently selected at each occurrence from hydrogen, heteroalkyl, alkyl, arylalkyl, heteroaryl alkyl, alkenyl, alkynyl, and, haloalkyl, heteroaryl, heterocycle, -alkyl-OR 8 , -alkyl-NR 8 R 9 , C(O)R 3 , S(O)R 3 , C(S)R 3 , and S(O)2R 3 .
  • R 8 and R 9 are independently selected at each occurrence from hydrogen, heteroalkyl, alkyl, arylalkyl, heteroarylalkyl, alkenyl, alkynyl, aryl, heteroaryl, and heterocycle.
  • the compound of Formula II has the structure of Formula II- A.
  • [MIFBM] and [IgGBM] are as defined herein.
  • a compound of Formula II-A havin the structure: wherein: [MIFBM/IgGBM] is a MIF or IgG Binding Moiety which binds respectively to circulating MIF or IgG in a subject, each of which is related to a disease state or condition and is to be removed by action of hepatocytes or other cells of the subject; [ASGPBM] is an asialoglycoprotein receptor binding moiety having the structure selected from each [CON] is an optional connector chemical moiety which, when present, connects the [LIN] to [CPBM] or to [ASGPBM]; [LIN] is [LINKER] or [LINKER-2], each of which is a chemical moiety having a valency from 1 to 15, which covalently attaches to one or more [ASGPBM] or [CPBM] groups, optionally through a [CON], wherein the [LIN] optionally itself contains one or more [CON] groups; Z B is absent, (CH 2 ) IM , C(O)-(CH 2 )
  • the ASGPR binding moieties can be any of the moieties described in: Reshitko, G. S., et al., “Synthesis and Evaluation of New Trivalent Ligands for Hepatocyte Targeting via the Asialoglycoprotein Receptor,” Bioconjugate Chem, doi: 10.1021/acs.bioconjchem.0c00202; Majouga, A. G., et al., “Identification of Novel Small- Molecule ASGP-R Ligands,” Current Drug Delivery, 2016, 13, 1303-1312, doi: 10.2174/1567201813666160719144651; Olshanova, A.
  • ASGPR binding moieties are illustrative and not intended to be limiting.
  • the ASGPR binding moiety can be a moiety having the structure of Ml, M2, M3, or M4, or a combination thereof.
  • X is independently at each occurrence O, NH, or S.
  • compounds of Formula I or Formula II can have one, two, or three ASGPR binding moieties with the structure of Ml, M2, M3, or M4.
  • ASGPR binding moieties Ml to M4 can be conjugated to any suitable [CON], [Linker], or [Linker-2] as described herein and in Congdon, M. D., et al., “Enhanced Binding and Reduced Immunogenicity of Glycoconjugates Prepared via Solid- State Photoactivation of Aliphatic Diazirine Carbohydrates,” Bioconjugate Chem. doi: 10.1021/acs.bioconjchem.0c00555.
  • the ASGPR binding moiety can be a moiety having the structure of M5 : , M5.
  • each R is independently at each occurrence R1 or R2, .
  • compounds of Formula I or Formula II contain an ASGPR binding moiety with the structure of M5.
  • each R in M5 is R 1 .
  • each R in M5 is R 2 .
  • ASGPR binding moiety M5 can be conjugated/bonded to any suitable [CON], [Linker], or [Linker-2] as described herein and in Reshitko, G.
  • the ASGPR binding moiety can be the galactose behenic acid ester-derived moiety M7: In the structure M7, Y is OH or NHAc. In various embodiments, the ASGPR binding moiety can be the agarose behenic acid ester-derived moiety M8:
  • ASGPR binding moieties M7 and M8 can be conjugated to any suitable [CON], [Linker], or [Linker-2] as described herein and in Dhawan, V., et al., “Polysaccharide conjugates surpass monosaccharide ligands in hepatospecific targeting - Synthesis and comparative in silico and in vitro assessment,” Carbohydrate Research 509 (2021) 108417, doi: 10.1016/j.carres.202L 108417.
  • the ASGPR binding moiety can be any of the compounds 2-
  • R is CILOAc, COOH, or CH2OH.
  • Compounds 2-18 can be conjugated/bonded to any suitable [CON], [Linker], or [Linker-2] as described herein and in Majouga, A. G., et al., “Identification of Novel Small-Molecule ASGP-R Ligands,” Current Drug Delivery, 2016, 13, 1303-1312, doi: 10.2174/1567201813666160719144651; Olshanova, A. S., et al., “Synthesis of a new betulinic acid glycoconjugate with N-acetyl-D-galactosamine for the targeted delivery to hepatocellular carcinoma cells,” Russian Chemical Bulletin, International Edition, Vol. 69, No. 1, pp.
  • Compounds 2-18 can be attached through any suitable reactive group contained therein. Without limitation, compounds 2-13 can be attached to a CON], [Linker], or [Linker-2] through or by reaction with at least one OH, NH, vinyl, alkynyl, amide, acid, ester, ketone, or aromatic halogen contained in compounds 2-18.
  • Suitable reaction modes for attaching compounds 2-18 to a [CON], [Linker], or [Linker-2] as described herein include, but are not limited to, substitution (e.g. alkylation of OH or NH groups), esterification (forming an ester), amidation (forming an amide), transesterification (exchanging one ester for another), transamidation (exchanging one amide for another), azide-alkyne cycloaddition, and other reactions capable of forming C-C, N-C, or O-C bonds with vinyl and alkynyl groups such as cycloadditions, aminations, oxidations, alkylations, rearrangement reactions (e.g. Claisen, Cope, etc.), and the like.
  • compositions herein which are presented to increase the solubility of the compound in saline for parenteral delivery or in the gastric juices of the patient’s gastrointestinal tract in order to promote dissolution and the bioavailability of the compounds.
  • Pharmaceutically acceptable salts include those derived from pharmaceutically acceptable inorganic or organic bases and acids. Suitable salts include those derived from alkali metals such as potassium and sodium, alkaline earth metals such as calcium, magnesium and ammonium salts, among numerous other acids well known in the pharmaceutical art.
  • sodium and potassium salts are used as neutralization salts of carboxylic acids and free acid phosphate containing compositions according to the present disclosure.
  • the term “salt” shall mean any salt consistent with the use of the compounds according to the present disclosure.
  • the term “salt” shall mean a pharmaceutically acceptable salt, consistent with the use of the compounds as pharmaceutical agents.
  • coadministration shall mean that at least two compounds or compositions are administered to the patient at the same time, such that effective amounts or concentrations of each of the two or more compounds may be found in the patient at a given point in time.
  • compounds according to the present disclosure may be co-administered to a patient at the same time, the term embraces both administration of two or more agents at the same time or at different times, provided that effective concentrations of all coadministered compounds or compositions are found in the subject at a given time.
  • Chimeric antibodyrecruiting compounds according to the present disclosure may be administered with one or more additional anti-cancer agents or other agents which are used to treat or ameliorate the symptoms of cancer, especially prostate cancer, including metastatic prostate cancer.
  • anticancer agent refers to a compound other than the chimeric compounds according to the present disclosure which may be used in combination with a compound according to the present disclosure for the treatment of cancer.
  • exemplary anticancer agents which may be coadministered in combination with one or more chimeric compounds according to the present disclosure include, for example, antimetabolites, inhibitors of topoisomerase I and II, alkylating agents and microtubule inhibitors (e.g., taxol), among others.
  • Exemplary anticancer compounds for use in the present disclosure may include everolimus, trabectedin, abraxane, TLK 286, AV-299, DN-101 , pazopanib, GSK690693, RTA 744, ON O91O.Na, AZD 6244 (ARRY-142886), AMN-107, TKI-258, GSK461364, AZD 1152, enzastaurin, vandetanib, ARQ-197, MK-0457, MLN8054, PHA-739358, R-763, AT-9263, a FLT-3 inhibitor, a VEGFR inhibitor, an EGFR TK inhibitor, an aurora kinase inhibitor, a PIK-1 modulator, a Bcl-2 inhibitor, an HD AC inhbitor, a c-MET inhibitor, a PARP inhibitor, a Cdk inhibitor, an EGFR TK inhibitor, an IGFR-TK inhibitor, an anti-HGF antibody, a PI3
  • a number of other agents may be coadministered with chimeric compounds according to the present disclosure in the treatment of cancer.
  • agents include active agents, minerals, vitamins and nutritional supplements which have shown some efficacy in inhibiting cancer tissue or its growth or are otherwise useful in the treatment of cancer.
  • active agents include active agents, minerals, vitamins and nutritional supplements which have shown some efficacy in inhibiting cancer tissue or its growth or are otherwise useful in the treatment of cancer.
  • curcumin turmeric
  • vitamin D green tea
  • omega-3 fatty acids including beta-sitosterol
  • compounds according to the present disclosure which contain a MIF binding moiety (MIFBM) or antibody binding moiety and ASGPR binding moiety selectively bind to liver cells and through that binding, facilitate the introduction of the MIFBM group into hepatocytes which bind the ASGPRBM selectively, where, the MIF protein, inside the hepatocyte is degraded and removed from circulation.
  • MIFBM MIF binding moiety
  • ASGPRBM ASGPR binding moiety
  • compositions comprising combinations of an effective amount of at least one compound disclosed herein, often a bi-functional chimeric compound (containing at least one MIFBM group or antibody binding moiety and at least one ASGPRBM) according to the present disclosure, and one or more of the compounds as otherwise described herein, all in effective amounts, in combination with a pharmaceutically effective amount of a carrier, additive or excipient, represents a further aspect of the present disclosure. These may be used in combination with at least one additional, optional anticancer agent as otherwise disclosed herein.
  • compositions of the present disclosure may be formulated in a conventional manner using one or more pharmaceutically acceptable carriers and may also be administered in controlled-release formulations.
  • Pharmaceutically acceptable carriers that may be used in these pharmaceutical compositions include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as prolamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene- polyoxypropylene-block polymers, polyethylene glycol and wool fat.
  • compositions of the present disclosure are administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir, among others.
  • parenteral as used herein includes subcutaneous, intravenous, intramuscular, intra-articular, intra-synovial, intrastemal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques.
  • the compositions are administered orally (including via intubation through the mouth or nose into the stomach), intraperitoneally or intravenously.
  • Sterile injectable forms of the compositions of this disclosure may be aqueous or oleaginous suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents.
  • the sterile injectable preparation may also be a sterile injectable solution or suspension in a nontoxic parenterally-acceptable diluent or solvent, for example as a solution in 1, 3 -butanediol.
  • the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution.
  • sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono- or di-glycerides.
  • Fatty acids such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions.
  • oils such as olive oil or castor oil, especially in their polyoxyethylated versions.
  • These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant, such as Ph. Helv or similar alcohol.
  • compositions of this disclosure may be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, aqueous suspensions or solutions.
  • carriers which are commonly used include lactose and corn starch.
  • Lubricating agents such as magnesium stearate, are also typically added.
  • useful diluents include lactose and dried corn starch.
  • aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring or coloring agents may also be added.
  • compositions of this disclosure may be administered in the form of suppositories for rectal administration.
  • suppositories for rectal administration.
  • suppositories can be prepared by mixing the agent with a suitable non-irritating excipient which is solid at room temperature but liquid at rectal temperature and therefore will melt in the rectum to release the drug.
  • suitable non-irritating excipient include cocoa butter, beeswax and polyethylene glycols.
  • compositions of this disclosure may also be administered topically, especially to treat skin cancers, psoriasis or other diseases which occur in or on the skin.
  • Suitable topical formulations are readily prepared for each of these areas or organs.
  • Topical application for the lower intestinal tract can be effected in a rectal suppository formulation (see above) or in a suitable enema formulation.
  • Topically-acceptable transdermal patches may also be used.
  • the pharmaceutical compositions may be formulated in a suitable ointment containing the active component suspended or dissolved in one or more carriers.
  • Carriers for topical administration of the compounds of this disclosure include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water.
  • the pharmaceutical compositions can be formulated in a suitable lotion or cream containing the active components suspended or dissolved in one or more pharmaceutically acceptable carriers.
  • suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2- octyl dodecanol, benzyl alcohol and water.
  • the pharmaceutical compositions may be formulated as micronized suspensions in isotonic, pH adjusted sterile saline, or as solutions in isotonic, pH adjusted sterile saline, either with our without a preservative such as benzylalkonium chloride.
  • the pharmaceutical compositions may be formulated in an ointment such as petrolatum.
  • compositions of this disclosure may also be administered by nasal aerosol or inhalation.
  • Such compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other conventional solubilizing or dispersing agents.
  • compositions are formulated to contain between about 0.05 milligram to about 1.5 grams, from 0.1 milligram to 1 gram, 0.5 milligrm to 750 milligrams, more often about 1 milligram to about 600 milligrams, and even more often about 10 milligrams to about 500 milligrams of active ingredient, alone or in combination with at least one additional compound which may be used to treat cancer, prostate cancer or metastatic prostate cancer or a secondary effect or condition thereof.
  • Methods of treating patients or subjects in need for a particular disease state or condition as otherwise described herein, especially cancer comprise administration of an effective amount of a pharmaceutical composition comprising therapeutic amounts of one or more of the novel compounds described herein and optionally at least one additional bioactive (e.g., anti-cancer, anti-inflammatory) agent.
  • a pharmaceutical composition comprising therapeutic amounts of one or more of the novel compounds described herein and optionally at least one additional bioactive (e.g., anti-cancer, anti-inflammatory) agent.
  • additional bioactive e.g., anti-cancer, anti-inflammatory
  • the amount of active ingredient(s) used in the methods of treatment of the instant disclosure that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated, the particular mode of administration.
  • compositions could be formulated so that a therapeutically effective dose of between about 0.01, 0.1, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90 or 100 mg/kg of patient/day or in some embodiments, greater than 100, 110, 120, 130, 140, 150, 160, 170, 180, 190 or 200 mg/kg of the novel compounds can be administered to a patient receiving these compositions.
  • a specific dosage and treatment regimen for any particular patient will depend upon a variety of factors, including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the judgment of the treating physician and the severity of the particular disease or condition being treated.
  • a patient or subject e.g. a human suffering from an autoimmune disease, an inflammatory disease or cancer can be treated by administering to the patient (subject) an effective amount of a chimeric/bi-functional compound according to the present disclosure including pharmaceutically acceptable salts, solvates or polymorphs, thereof optionally in a pharmaceutically acceptable carrier or diluent, either alone, or in combination with other known pharmaceutical agents, exemplary agents which can assist in treating autoimmune and/or inflammatory diseases or cancer, including metastatic cancer or recurrent cancer or ameliorating the secondary effects and/or symptoms assocated with these disease states and/or conditions.
  • This treatment can also be administered in conjunction with other conventional therapies, such as radiation treatment or surgery for cancer.
  • present compounds alone or in combination with other agents as described herein, can be administered by any appropriate route, for example, orally, parenterally, intravenously, intradermally, subcutaneously, or topically, in liquid, cream, gel, or solid form, or by aerosol form.
  • the active compound is included in the pharmaceutically acceptable carrier or diluent in an amount sufficient to deliver to a patient a therapeutically effective amount for the desired indication, without causing serious toxic effects in the patient treated.
  • An exemplary dose of the active compound for all of the herein-mentioned conditions is in the range from about 10 ng/kg to 300 mg/kg, such as 0.1 to 100 mg/kg per day, such as 0.5 to about 25 mg per kilogram body weight of the recipient/patient per day. Some exemplary dosages will range from about 0.01-3% wt/wt in a suitable carrier.
  • the compound is conveniently administered in any suitable unit dosage form, including but not limited to one containing less than Img, 1 mg to 3000 mg, such as 5 to 500 mg of active ingredient per unit dosage form.
  • An oral dosage of about 25-500 mg is often convenient.
  • the active ingredient is sometimes administered to achieve peak plasma concentrations of the active compound of about 0.00001-30 mM, such as about 0.1-30 pM. This may be achieved, for example, by the intravenous injection of a solution or formulation of the active ingredient, optionally in saline, or an aqueous medium or administered as a bolus of the active ingredient. Oral administration is also appropriate to generate effective plasma concentrations of active agent.
  • the concentration of active compound in the drug composition will depend on absorption, distribution, inactivation, and excretion rates of the drug as well as other factors known to those of skill in the art. It is to be noted that dosage values will also vary with the severity of the condition to be alleviated. It is to be further understood that for any particular subject, specific dosage regimens should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions, and that the concentration ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed composition.
  • the active ingredient may be administered at once, or may be divided into a number of smaller doses to be administered at varying intervals of time.
  • Oral compositions will generally include an inert diluent or an edible carrier. They may be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the active compound or its prodrug derivative can be incorporated with excipients and used in the form of tablets, troches, or capsules. Pharmaceutically compatible binding agents, and/or adjuvant materials can be included as part of the composition.
  • the tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a dispersing agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterotes; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.
  • a binder such as microcrystalline cellulose, gum tragacanth or gelatin
  • an excipient such as starch or lactose, a dispersing agent such as alginic acid, Primogel, or corn starch
  • a lubricant such as magnesium stearate or Sterotes
  • a glidant such as colloidal silicon dioxide
  • the active compound or pharmaceutically acceptable salt thereof can be administered as a component of an elixir, suspension, syrup, wafer, chewing gum or the like.
  • a syrup may contain, in addition to the active compounds, sucrose as a sweetening agent and certain preservatives, dyes and colorings and flavors
  • the active compound or pharmaceutically acceptable salts thereof can also be mixed with other active materials that do not impair the desired action, or with materials that supplement the desired action, such as other anticancer agents, anti-inflammatory agents, immunosuppressants, antibiotics, antifungals, or antiviral compounds.
  • one or more chimeric/bi-functional MIF binding compound according to the present disclosure is coadministered with another anticancer agent and/or another bioactive agent, as otherwise described herein.
  • Solutions or suspensions used for parenteral, intradermal, subcutaneous, or topical application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose.
  • the parental preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.
  • PBS physiological saline or phosphate buffered saline
  • the active compounds are prepared with carriers that will protect the compound against rapid elimination from the body, such as a controlled and/or sustained release formulation, including implants and microencapsulated delivery systems.
  • a controlled and/or sustained release formulation including implants and microencapsulated delivery systems.
  • Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Methods for preparation of such formulations will be apparent to those skilled in the art.
  • Liposomal suspensions or cholestosomes may also be pharmaceutically acceptable carriers. These may be prepared according to methods known to those skilled in the art, for example, as described in U.S. Pat. No. 4,522,811 (which is incorporated herein by reference in its entirety).
  • liposome formulations may be prepared by dissolving appropriate lipid(s) (such as stearoyl phosphatidyl ethanolamine, stearoyl phosphatidyl choline, arachadoyl phosphatidyl choline, and cholesterol) in an inorganic solvent that is then evaporated, leaving behind a thin film of dried lipid on the surface of the container. An aqueous solution of the active compound are then introduced into the container. The container is then swirled by hand to free lipid material from the sides of the container and to disperse lipid aggregates, thereby forming the liposomal suspension.
  • appropriate lipid(s) such as stearoyl phosphatidyl ethanolamine,
  • FIGURES 1, 7, and 13 attached hereto identifies exemplary compounds according to the present disclosure which exhibit activity in binding to and reducing and/or eliminating unwanted circulating proteins for therapeutic and/or diagnostic purposes.
  • These compounds are based upon an MIF, anti-DNP IgG, or IgG binding moiety to which is covalently attached an ASPGR group such as GN3 or AcF3-3 group through a linker which contains from 1 to 100 ethylene glycol groups, more often from 1 to 15 ethylene glycol groups, from 1 to 10 ethylene glycol groups, often from 2 to 10 ethylene glycol groups which are optionally attached through a [CON] group, such as a 1,2,3-triazole or other [CON] group as described herein.
  • ASPGR group such as GN3 or AcF3-3 group
  • linker which contains from 1 to 100 ethylene glycol groups, more often from 1 to 15 ethylene glycol groups, from 1 to 10 ethylene glycol groups, often from 2 to 10 ethylene glycol groups which are optionally attached through a [
  • FIGURE 16 shows the synthesis of azide/amide carboxylic end capped PEG linker intermediates which may be condensed onto an alkynyl precursor (e.g. NVS alkyne precursor of Figure 5) to provide carboxylic acid capped intermediate which can be used to provide bifunctional molecules.
  • alkynyl precursor e.g. NVS alkyne precursor of Figure 5
  • FIGURE 17 describes a general method for conversion of PEG molecules into hydroxyl azides.
  • the PEG compound is tosylated (TsCl, DCM, in the presence of base) at reduced temperature and further reacted with sodium azide at elevated temperature in a non- nucleophilic solvent.
  • the final azidoalcohol is used in subsequent figures.
  • FIGURE 18 describes the synthesis of a mesylated azide from a starting PEG molecule employing the same synthetic steps to reach the intermediate azido alcohol. This is then treated with MsCl in pyridine to afford the final compound.
  • FIGURE 19 shows the synthesis of the GalNAc ASGPR ligand linked through PEG to a terminating amine.
  • Pentaacetyl galactosamine is reacted with TMSOTf at elevated temperature in DCE to produce a bicyclic intermediate, which is then reacted with an azido alcohol to give an azide intermediate (TMSOTf, DCE).
  • TMSOTf azide intermediate
  • This molecule is then subjected to a Staudinger reduction to give an amine which is used in subsequent figures.
  • FIGURE 20 shows the synthesis of a higher affinity bicylic ASGPR ligand.
  • Galactose pentaacetate is treated with HBr/AcOH to give the brominated intermediate, which is treated with Zn and CuSO4 (water/ AcOH) to give the galactal.
  • This is treated with ammonium cerium nitrate and sodium azide at reduced temperature (MeCN) to give the disubstituted intermediate compound.
  • This is then treated with strong base (NaOMe/MeOH) to give the triol azide intermediate.
  • This compound is silylated completely (TMSCl/pyr) then the primary alcohol is deprotected (potassium carbonate, MeOH, lowered temperature) and oxidized (Dess-Martin Periodinane, DCM).
  • Treatment with strong base (NaOEt/HOEt) and paraformaldehyde gives the tetraol intermediate, which is cyclized in strong acid (H2SO4/water) to give the bicyclic azide ligand.
  • FIGURE 21 shows the synthesis of a trifluoro-acetate derivative of the bicyclic ASGPR ligand.
  • the triol azide is reduced (Pd/C, MeOH) to give the intermediate amine, which is then peracylated with trifuloroacetic anhydride.
  • the esters are hydrolyzed with strong base (NaOMe/HOMe) to give the intermediate amide, which is protected using dimethoxypropane in the presence of camphorsulfonic acid in DMF at elevated temperature. This is then reacted a mesylated azdio alcohol in the presence of strong base (NaH/DMF) to give an intermediate azide that is reduced (Lindlar’s catalyst, MeOH) to give the final amine.
  • FIGURE 22 shows the synthesis of the MIF -targeting linker to a monovalent linker, which is synthesized through analagous methods as described in a previous figure.
  • the boc- protected methyl ester is deprotected with TFA in DCM, then coupled to the MIF-targeting carboxylic acid (HBTU, DIPEA, DMF). Subsequent hydrolysis with strong base (NaOH/dioxane/H2O) gives the MIF-targeting carboxylic acid.
  • FIGURE 23 shows the synthesis of the di-carboxylic acid MIF targeting motif, which is synthesized as described in previous figures.
  • FIGURE 24 shows the synthesis of a tris base-derived trivalent linker.
  • Tris base is treated with di-t-butyl dicarbonate in the presence of base to give the boc protected triol, which is then reacted with acrylonitrile in the presence of base (dioxane/H2O) to give a trinitrile intermediate. This is then converted to the methyl ester through treatment with strong acid in methanol.
  • the amine is then reacted with Cbz-glycine through a DCC- mediated amide formation, and deprotected to give a tricarboxylic acid that is used in subsequent figures.
  • FIGURE 25 describes the synthesis of an ASGPR-targeting moiety employing three GalNAc ASGPR ligands.
  • the tricarboxylic acid is reacted with amine-terminated protected GalNAc (amide bond formation in the presence of HBTU and DIPEA), then deprotected by reduction (Pd/C, solvent) and treatment with strong base (NaOMe/MeOH).
  • FIGURE 26 shows the synthesis of the tri-carboxylic acid MIF targeting motif, which is synthesized as described in previous figures.
  • FIGURE 27 shows the synthesis of the MIF NVS alkyne precursor which can be reacted with an azido reactant containing a carboxylic acid (as set forth in subsequent figures) to provide MIF-NVS-carboxylic acid capped reactants to produce the bifunctional compounds.
  • FIGURE 28 shows the synthesis of the MIF-targeting moiety terminating in a carboxylic acid.
  • 2-chloroquinolin-6-ol is reacted with ethyl 4-bromobutanoate in the presence of base (DMF, elevated temperature) to give an aryl chloride that then undergoes Sonogashira coupling at evelvated temperature with ethynyltrimethylsilane.
  • the intermediate silylated compound is deprotected with TBAF (DCM/THF).
  • a click reaction then forms a triazole between the alkyne intermediate and in situ synthesized 4-azido-2-fluorophenol to give an ethyl ester intermediate that is hydrolyzed with strong base (NaOH/dioxane) to give the carboxylic acid that is used in subsequent figures.
  • FIGURE 29 describes the synthesis of the bifunctional molecule MIF-NVS-PEGn- GN3 through HBTU-mediated coupling in DMF of the ASPGR-targeting amine and the MIF targeting carboxylic acid prepared by first forming the MIF-targeting carboxylic acid by condensing the reactant azido PEG-carboxylic acid onto the MIF moiety containing a alkyne terminated PEG group.
  • FIGURE 30 describes the synthesis of bifunctional molecules MIF-GN3 and MIF- PEGn-GN3 through HATU-mediated coupling (DMF, DIPEA) of ASGPR-targeting amine and MIF-targeting carboxylic acid.
  • FIGURE 31 describes the synthesis of the bifunctional molecule targeting MIF and ASGPR, containing one bicyclic ASGPR AcF3 ligands.
  • MIF-binding mono-carboxylic acid is treated with HBTU, DIPEA, the amine terminated ligand, and DMF to give the amide, which is then deprotected with IM HC1 to give the final compound.
  • FIGURE 32 describes the synthesis of the bifunctional molecule targeting MIF and ASGPR, containing two bicyclic ASGPr ligands. It is synthesized as described above.
  • FIGURE 33 describes the synthesis of the bifunctional molecule targeting MIF and ASGPR, containing three bicyclic ASGPr ligands. It is synthesized as described above.
  • FIGURE 34 shows the synthesis of DNP-GN3. 2,4-dinitro chlorobenzene was treated with an amino carboxylic acid in the presence of weak base to give the di-nitro analine carboxylic acid intermediate. Further steps were carried out as described for previous molecules.
  • FIGURE 35 shows the synthesis of DNP-AcF3-3, which was carried out with methods analogous previous compounds.
  • FIGURE 36 shows the synthetic scheme used to obtain IBA-GN3.
  • Pentaethylene glycol was treated with tosyl chloride in the presence of base to give the mono-tosylated alcohol, which was then treated with sodium azide at elevated temperature to give the azidoalcohol.
  • This compound was then oxidized using Jones reagent, then reduced with Palladium on carbon under hydrogen atmosphere to give a carboxylic acid-amine.
  • indole butyric acid was treated with N-hydroxysuccinimide, EDC, and DIPEA to give the NHS-ester indole, which was then reacted with the above carboxylic acid-amine.
  • FIGURE 37 shows the synthesis of triazine-GN3.
  • Cyanuric chloride was treated with (4-(methoxycarbonyl)phenyl)methanaminium in THF and diisopropylethlamine at -78° C to give the mono-substituted product. This was then treated with cyclohexylmethanamine at room temperature to afford the second substitution. The final substitution was accomplished under elevated temperature with (lS,2S,4R)-bicyclo[2.2.1]heptan-2-amine to give the tri substituted triazine.
  • FIGURE 38 shows the synthetic scheme used to access FcIII-GN3.
  • the hexynyl peptide was prepared using standard solid phase peptide synthesis techniques. The peptide was removed from Rink resin using Reagent L, then oxidized using ammonium bicarbonate buffer (pH8-9) in MeOH under air to give the cyclic peptide. The peptide was reacted with GN3-azide, which was described previously, to give the product triazole FcIII-GN3.
  • FIGURE 39 shows the synthetic scheme used to access FcIII-4c-GN3, which was accomplished using methods described above.
  • FIGURES 40-43 describe the synthesis of bifunctional molecules targeting MIF and ASGPr, containing three bicyclic ASGPR ligands with different substitutions on the 2-amine of the sugar. They are synthesized through analagous methods described above as set forth in the attached figures.
  • FIGURE 44 shows the synthesis of compound MIF-18-3.
  • Tri-acyl galactal was deprotected with ammonia in methanol, then tri-benzyl protected with benzylbromide in the presence of base.
  • the alkene was hydrolyzed overnight with HC1 in THF/H2O, then oxidized with PCC to give an aldehyde.
  • Sodium azide was then added alpha to the carbonyl with KHMDS and TIBSN3 at lowered temperature.
  • the intermediated was then treated with p- OMePhMgBr in THF and toluene to give an intermediate alcohol, which was then reduced using Et3SiH in the presence of BF3-Et2O at reduced temperature.
  • FIGURE 45 shows the synthesis of compound MIF-31-3.
  • Galactosamine hydrochloride was fully protected with acetic anhydride, then treated with allyl alcohol in the presence of BF3 ehtrate to give the allyl intermediate.
  • Treatment with pivaloyl chloride in pyridine gave a di-Piv protected intermediate, which was treated with triflic anhydride and subsequently subjected to hydrolysis in water at elevated temperature.
  • the pivaloyl groups were removed by treatment with NaOMe in MeOH to give the allyl triol intermediate. Subsequent steps were performed as described for previous molecules.
  • FIGURE 46 shows the synthesis of compound MIF-15-3, which was synthesized using procedures analogous to compounds described above.
  • FIGURE 47 shows the synthesis of compound MIF-19-3.
  • the molecule is synthesized through a late stage triazole-forming click reaction between the triazide and propiolic acid in methanol in the presence of THPTA, copper sulfate, water, and sodium ascorbate. All other reactions are performed as described above.
  • FIGURE 48 shows the synthesis of compound MIF- 16-3, which was synthesized using procedures analogous to compounds described above.
  • FIGURE 49 shows the synthesis of compound MIF-20-3, which was synthesized using procedures analogous to compounds described above.
  • FIGURE 50 shows the synthesis of compound MIF-14-3, which was synthesized using procedures analogous to compounds described above.
  • FIGURE 51 shows the synthesis of compound MIF-21-3, which was synthesized using procedures analogous to compounds described above.
  • FIGURES 52-66 show the synthesis of a number of MIF-binding compounds with various ASGPRBM moieties. These are synthesized through methods analogous to those laid out above.
  • Proper protein section and turnover is a necessary process for maintaining homeostasis.
  • Newly synthesized proteins targeted for secretion are first trafficked to the endoplasmic reticulum, where they are post-translationally modified with N-linked glycan chains terminating in sialic acids(l).
  • terminal sialic acid residues are removed by circulating endogenous glycosydases(2).
  • This natural protein aging process unmasks galactose and N-acetylgalactose (GalNAc) residues, which bind the asialoglycoprotein receptor (ASGPR) on the surface of hepatocytes(3-5).
  • GalNAc N-acetylgalactose
  • the ASGPR is a C-type lectin that removes aged circulating proteins with exposed GalNAc residues from circulation by trafficking them to lysosomes. Multiple galactose or GalNAc residues displayed on the protein surface are necessary for high-affinity binding to - and subsequent endocytosis by - ASGPR(6, 7). Once these proteins are endocytosed, they are released from the ASGPR through depletion of calcium from the endosome and changes in binding site amino acid protonation changes due to a decrease in pH (12); the ASGPR is recycled back to the hepatocyte surface(13). Endocytosed proteins are trafficked to late endosomes, which are fused with lysosomes.
  • Lysosomal proteases then degrade endocytosed proteins, permanently removing them from circulation(14).
  • Non-glycosylated proteins are not known to be natural target for the ASGPR.
  • MIF macrophage inhibitory factor
  • Genetic depletion or antibody neutralization of MIF has been shown to have positive results in models of sepsis(16), multiple schlorosis(17), rheumatoid arthritis(18), and burn recovery(21).
  • a bifunctional molecule for degrading cicrulating MIF that takes advantage of ASGPR as an entryway for proteins into the endosomal-lysosomal degradation pathway was proposed.
  • the bicyclic ASGPR-binding molecules in MIF-AcF2 and MIF- AcF3 have been reported previously as high affinity binders for the ASGPR (22, 23).
  • Example 1 exemplary compounds according to some embodiments were tested to determine their biological activity. Active compounds are shown in FIGURES 1, 7 and 13 hereof. The results of the biological experiments are described herein below.
  • FIGURE 1 shows exemplary compounds according to the present disclosure. Note that the figure discloses compound 3w (negative control for MIF inhibition), MIF-NVS- PEGnGN3, MIFGN3, MIF-PEGnGN3, MIF-AcF3-l, MIF-AcF3-2 and MIF-AcF3-3. Note that n in the PEG linker sometimes ranges from 1-12, such as 1 to 10, 2 to 8, 2 to 6, 2 to 5 or 1, 2, 3 or 4.
  • FIGURE 2 A. fluorescence polarization data of MIF-FITC binding to human MIF indicates that the MIF-binding moiety of the present disclosure binds MIF.
  • bifunctional molecules were able to deplete human MIF from the supernatant of culture HepG2 cells.
  • human MIF 100 nM was added to cell culture media in the presence of negative control MIF inhibitor 3w as well as bifunctional molecules MIF-NVS-PEGn-GN3, MIF- GN3, MIF-PEGn-Gn3, MIF-AcF3-l, MIF-AcF3-2, and MIF-AcF3-3. All molecules utilized a known MIF-binding ligand.
  • All molecules utilized a known MIF-binding ligand.
  • Experiments were performed in 96 well plates (approximate surface area .3 cm 2 ).
  • HepG2 cells were grown to 90% confluency in RPMI media, then washed with PBS (2x) and treated with serum-free media (optimem + .1% BSA, + Pen/Strep) containing 100 nM huMIF (Cayman Chemical) and compounds (when applicable).
  • Compounds were diluted from ImM stock solutions in DMSO. After 24 hours, a sample of the supernatant (2 uL) was collected, diluted 1 TOO, and analyzed for MIF content by sandwich ELISA (and incubated for 24 hours in the presence or absence of compound). Remaining MIF levels were determined by sandwich ELISA (biolegend monoclonal anti- MIF and biotinylated anti-MIF antibodies). Data represents the average of at least 3 biological replicates, and error bars represent a standard deviation. After 24 hours, up to 95.3% of the MIF had been depleted from cell culture media (in the case of MIF-AcF3-3).
  • FIGURE 4 shows the results of an experiment to determine whether or not MIF internalized by HepG2 cells is trafficked to lysosomes.
  • cells were incubated with rhuMIF (Cayman) at a concentration of 100 nM with 200 nM MIF-GN3. After 12 hours, cells were fixed with formaldehyde, permeabilized, and probed with anti- Lamp2 antibody (mouse monoclonal, Abeam), polyclonal rabbit anti-MIF antibody (Thermo) and with Alexa-488 labeled anti-mouse antibody and Alexa 568-labeled anti-rabbit antibody, evidencing internalization in lysosomes.
  • Lamp2 antibody mouse monoclonal, Abeam
  • polyclonal rabbit anti-MIF antibody Thermo
  • Alexa-488 labeled anti-mouse antibody and Alexa 568-labeled anti-rabbit antibody evidencing internalization in lysosomes.
  • FIGURE 5 shows that MIF-GN3 mediates the depletion of injected human MIF from mice.
  • Human MIF has a half-life of approximately 40 minutes in mice.
  • human recombinant MIF (Cayman chemical) was co-injected into mice with an anti-DNP IgG, which was used as an injection positive control.
  • nude mice were injected with 5 pg recombinant human MIF and 200pg anti-DNP IgG as an injection control (FIGURE 4).
  • MIF-GN3 was then injected at the concentration shown and blood drawn every twenty minutes over the course of two hours.
  • Serum was diluted 1 : 100 and analyzed for MIF content by sandwich ELISA (biolegend monoclonal anti-MIF and biotinylated anti-MIF antibodies).
  • the levels of the injected IgG were not significantly different between testing groups. In the mice treated with MIF-GN3, a moderate increase in huMIF levels up to 20 ng/ml was seen, while in mice injected with PBS negative control, serum levels of up to 150 ng/ml were observed, evidencing a substantial decrease in huMIF levels as a consequence of the administration of MIF-GN3.
  • FIGURE 6 shows that MIF-GN3 is able to delay tumor growth in a mouse model of prostate cancer.
  • nude mice were engrafted with PC3 human prostate cancer cells. Treatment was then initiated immediately with either a non-bifunctional MIF inhibitor (3w), an anti-MIF antibody, or MIF-GN3.
  • MIF-GN3 showed a slowing of tumor growth over the course of the experiment, comparable to the MIF-neutralizing antibody. 3w did not inhibit tumor growth, validating the necessity of degrading MIF for therapeutic efficacy.
  • FIGURE 7 shows molecules DNP-GN3 and DNP-AcF3-3, which are bifunctional molecules that bind to anti-DNP IgG and ASGPR. These compounds were used in several of the experiments as described below.
  • FIGURE 8 shows that DNP-GN3 and DNP-AcF3-3 mediate the formation of a ternary complex between HepG2 cells and anti-DNP, thus validating the bifunctional character of the molecules.
  • ASGPR-expressing HepG2 cells were incubated with bifunctional molecules and alexa-488 labeled anti-DNP (Thermo).
  • the readout is mean fluorescence intensity of the cell population. Fluorescence was measured using a flow cytometer.
  • results presented in FIGURE 9 show that DNP-GN3 and DNP-AcF3-3 mediate the uptake of alexa 488-labeled anti-DNP by HepG2 cells.
  • the assay carried out in this experiment was as is described above for MIF uptake. Readout is percentage of Alexa 488-positive cells after 6 hours. Fluorescence was measured using a flow cytometer.
  • FIGURE 10 shows that DNP-GN3 and DNP-AcF3-3 mediate the localization of alexa 568 labeled anti-DNP to late endosomes and lysosomes. This experiment was carried out as described above for the MIF colocalization studies.
  • FIGURE 11 The experimental results presented in FIGURE 11 show that DNP-AcF3-3 mediates the degradation of alexa 488-labeled anti-DNP in HepG2 cells.
  • cells were incubated with 1 uM alexa 488-labeled anti-DNP (Thermo) and 200 nM DNP-AcF3-3.
  • Cells were lysed (RIP A in PBS, containing protease inhibitors) at the given time and assayed by SDS-PAGE gel. Readout is fluorescence of protein fragments.
  • FIGURE 12 evidence that DNP-GN3 mediates the depletion of anti-DNP from mouse serum. Mice were injected with anti-DNP on day 0, then treated with the given compounds each day for 6 days. Serum IgG levels were measured by ELISA. DNP-(OH)3 is used as a non-bifunctional control molecule.
  • FIGURE 13 shows the structures of IgG-degrading molecules IBA-GN3, Triazine- GN3, FcIII-GN3, and FcIII-4c-GN3.
  • FIGURE 14 shows that FcIII-GN3 mediates the uptake of human IgG into HepG2 cells. This experiment was performed as described above.
  • FIGURE 15 shows that FcIII-GN3 mediates the localization of IgG to late endosomes in HepG2 cells. Experiment performed as described above.
  • Example 2-1 MIF binding molecule ( Figure 13)
  • Example 2-1-1 MIF-1 2-chloroquinolin-6-ol (1.00 g, 5.57 mmol) and K 2 CO 3 (1.53 g, 11.1 mmol, 2.0 eq) were dissolved in DMF (20 mL).
  • TMSOTf (55 uL, 67.5 mg, .304 mmol, .5 eq) was then added to the mixture, and the reaction stirred overnight.
  • the mixture was diluted into DCM, washed with 1M sodium bicarbonate (1x) and water (1x), then dried over magnesium concentrate and concentrated.
  • the curde oil was purified on silica gel (50-100% EtoAc in DCM) to give compound 67 (245 mg, .486 mmol) in 80.1% yield.
  • Triphenylphosphine (1.40 g, 5.35 mmol, 1.5 eq) and water (257 uL, 14.28 mmol, 4 eq) were then added and the reaction stirred at room temperature under nitrogen for 36 hours. The solvent was removed and the crude product used in the next step without further purification.
  • Example 2-4. Tris Valent Glycine (Figure 16) Tris base (5.00 g, 41.3 mmol) was dissolved in dichloromethane (80 mL) and trimethylamine (20 mL). Di-tert-butyl dicarbonate (10.81 g, 49.6 mmol, 1.2 eq) was then added, and the reaction stirred for 4 hours.
  • Tetraethylene glycol (50.0 g, 258 mmol) was dissolved in THF (1 mL), cooled to 0°, and stirred. NaOH (1.65 g, 41.3 mmol, 1.6eq) in water (1 mL) was then added, followed by the dropwise addition of p-toluenesulfonyl chloride (4.92 g, 25.8 mmol, 1 eq) in THF (3 mL). The reaction mixture was stirred at 0° for 4 hours, then diluted into DCM. The organic layer was washed with ice-cold water (2x), brine (lx), and dried over sodium sulfate to give compound 16 (8.84 g, 25.4 mmol, 99% yield), which was used in further steps without purification.
  • Methanesulfonyl chloride (3.14 g, 27.4 mmol, 1.2 eq) was then added and the reaction stirred for six hours under nitrogen. The mixture was then diluted into ethyl acetate, washed with water (3x), .5M HCl (2x), saturated sodium bicarbonate (1x), and brine (1x), dried over sodium sulfate, and evaporated to give compound 18 (5.71 g, 19.2 mmol) in 84% yield.
  • Trimetylsilylchloride (10.43 mL, 8.929 g, 82.18 mmol, 3.6 eq) was added dropwise and the mixture stirred for 6 hours. The reaction was diluted into ethyl acetate and washed with water (2x) and brine (1x). The organic layer was dried over sodium sulfate and evaporated to give the tri-TMS intermediate. Residual pyridine was removed by coevaporating with toluene (3x). The intermediate was taken up into dry MeOH (45 mL) and cooled to 0o before potassium carbonate (40 mg) was added. The reaction was closely monitored over 1.5 hours and quenched with acetic acid (17 uL) once TLC showed complete consumption of starting material.
  • Example 2-9 Bicyclic ASGPR ligand CF3 (Figure 21) Compound (400 mg, 1.84 mmol) was dissolved in methanol (30 mL) and the reaction flask purged with nitrogen. Lindlar’s catalyst (40.0 mg, 10 wt%) was then added, and the reaction mixture stirred under a 1 atm hydrogen atmosphere (balloon) for 6 hours. The reaction was filtered over celite and evaporated to give compound 10 (351 mg, 1.84 mmol) in quantitative yield, which was used in the next reaction without further purification.

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Abstract

Described herein is a bi-functional compound for removing macrophage migration inhibitory factor (MIF) or immunoglubin G (IgG). Further described herein is a pharmaceutical composition which comprise these bi-functional compounds. Further described herein is a method for treating disease states and/or conditions with the compounds or the composition. The disease states and/or conditions are mediated through MIF/IgG or where MIF/IgG is a contributing factor to the development and perpetuation of diseases and/or conditions, such as autoimmune diseases and cancer, among others.

Description

TITLE
BI-FUNCTIONAL MOLECULES TO DEGRADE CIRCULATING PROTEINS
CROSS-REFERENCE TO RELATED APPLICATIONS
This application claims priority to U.S. Application No. 17/695,259, filed March 15, 2022, which application is incorporated herein by reference in its entirety.
STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH
This invention was made with government support under GM067543 awarded by National Institutes of Health and under W81XWH- 13-1 -0062 awarded by the United States Army Medical Research and Material Command. The government has certain rights in the invention.
SEQUENCE LISTING
The instant application contains a Sequence Listing which has been submitted in ASCII format via EFS-Web and is hereby incorporated by reference in its entirety. Said ASCII copy, created on March 12, 2023, is named 47162-7240WO2_Sequence Listing. xml and is 30.8 kilobytes in size.
BACKGROUND
MIF is a proinflammatory pluripotent cytokine which contributes to the development and perpetuation of many diseases. These include atherosclerosis, rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), sepsis, inflammatory bowel disease (IBD), among others. In addition, MIF has been shown to participate in the progression of various cancers, including colon cancer, prostate cancer, breast cancer, lung cancer, cervical and adenocarcinoma, among others. In pre-clinical models, inhibition of MIF has been shown to attenuate symptoms and delay disease progression in the above-mentioned diseases.
MIF and IgG significantly contribute to the pathologic process of a number of diseases. MIF neutralizing antibodies and small molecule MIF inhibitors have been developed, where IgG is upregulated in autoimmune and other diseases and causes untoward effects. Because MIF acts through various pathways, a MIF inhibitor is sometimes required to be able to interrupt multiple protein-protein interactions (PPIs), posing a challenge for drug development. In the case of IgG, the removal of IgG molecules which are upregulated and contributing to autoimmune and/or other diseases is sometimes useful to inhibit and/or ameliorate the effects of the disease state. Therefore, a novel approach where MIF or IgG can be degraded as well as inhibited can overcome this difficulty and potentially provide a more robust therapeutic response. Encouraged by preliminary results with bi-functional MIF and IgG degraders, the present inventors decided to further test them in clinical-relevant in vivo models and potentially develop them into therapeutics that target MIF or IgG in diseases. They also intend to use this basic bifunctional molecule strategy to target other circulating proteins of interest to degradation via the ASGPr.
Therefore, there is a need for molecules that can eliminate MIP and/or IgG. The present disclosure addresses this need.
BRIEF SUMMARY
In some aspects, the present disclosure is directed to bifunctional small molecules which can be used to inhibit and remove MIF or IgG. In some embodiments, the present disclosure aims to establish a general small molecule strategy to target the selective degradation of MIF or IgG, which contributes to the development and perpetuation of many diseases. In some embodiments, the bifunctional molecule construct contains a MIF binding motif derived from known small molecule ligands of MIF of a IgG binding motif which comprises a ligand which binds IgG. These moieties are referred to herein generically as macrophage migration inhibitory factor binding moiety (MIFBM) or alternatively, immunoglobulin G binding moiety (IgGBM). In some embodiments, the other end of the bifunctional molecule is a motif that binds to hepatocyte asialoglycoprotein receptor (ASGPr), which is a triantennary N-acetylgalactosamine or other moiety as described herein. The instant specification sometimes refer to these moieties generically as asialoglycoprotein receptor binding moiety (ASGPRBM). The motifs which are used to provide bifunctional compounds according to some embodiments, (MIFBM or IgGBM) and (ASGPRBM), are covalently linked via a linker such as a polyethylene glycol (PEG) or other linker as described herein with adjustable length or other linker as described herein, which optionally contains one or more connector molecule(s) which connects the linker to the MIFBM or IgGBM and/or the ASGPRBM.
In some embodiments, thje bifunctional compounds selectively bind to MIF or IgG in circulation and form a protein complex. When this protein complex passes through the liver, the asialoglycoprotein receptor binding moiety of the molecule (ASGPRBM), such as a triantennary N-Acetylgalactosamine or other motif as described herein, will engage the endo- lysosomal pathway of hepatocytes through the ASGPr. As a consequence of this mechanism, MIF or IgG is eliminated from circulation by hepatocytes, thus resulting in lowered levels of MIF or IgG with the result being that corresponding disease symptoms are attenuated and/or eliminated from a patient or subject administered the present compounds. In certain instances, the MIF is substantially reduced or eliminated or the compromising IgG substantially reduced, resulting in substantially reduced symptoms or even a cure or elimination of the disease state or condition.
The approach pursuant to the present disclosure is inherently advantageous compared to the classical antibody -based strategy to target MIF or IgG of the prior art. The small molecule based approach of the current disclosure overcomes the limitations of traditional antibody-based strategies, including lack of oral bioavailability, low-temperature storage requirements, immunogenicity, and high-cost.
Furthermore, the compounds and methods of the present disclosure are expected to have a more lasting effect compared to the conventional inhibitory approach because MIF or IgG is substantially reduced or eliminated by degradation inside hepatocytes rather than simply inhibited by reversibly blocking the protein-receptor interaction. The bifunctional molecule construct pursuant to the present disclosure is also versatile in the sense that different disease states and/or conditions can be targeted by inhibiting and degrading MIF or IgG. Thus, previously discovered non-inhibitory protein binders, presently of little value in therapy, can be potentially therapeutically useful in these small molecules.
In certain embodiments, the present disclosure is directed to compounds which are useful for removing circulating proteins which are associated with a disease state or condition in a patient or subject according to the general chemical structure:
Figure imgf000005_0001
wherein [MIFBM/IgGBM] is a MIF or IgG binding moiety which binds respectively to circulating MIF or IgG, which are related to a disease state and/or condition and is to be removed by the action of hepatocytes on circulating protein (in some embodiments, the compounds selectively bind to MIF or IgG in plasma);
[ASGPRBM] is a binding moiety which binds to hepatocytes through asialoglycoprotein receptors which are on the surface of hepatocytes, such as in a patient or subject; each [CON] is an optional connector chemical moiety which, when present, connects directly to [MIFBM/IgGBM] or to [ASGPRBM] or connects the [LINKER] to [MIFBM/IgGBM] or to [ASGPRBM] and
[LINKER] is a chemical moiety having a valency ranging from 1 to 15, such as ranging from 1 to 10, ranging from 1 to 5 or 1, 2 or 3, which covalently attaches to one or more [ASGPRM] and/or [MIFBM/IgGBM] group, optionally through a [CON], including a [MULTICON] group, wherein said [LINKER] optionally itself contains one or more [CON] or [MULTICON] group(s); k’ is an integer ranging from 1 to 15, such as ranging from 1 to 10, ranging from 1 to 5, ranging from 1 to 3 or 1, 2 or 3; j’ is an integer ranging from 1 to 15, such as ranging from 1 to 10, ranging from 1 to 5, ranging from 1 to 3 or 1, 2 or 3; h and h’ are each independently an integer ranging from 0 to 15, such as ranging from 1 to 15, ranging from 1 to 10, ranging from 1 to 5, ranging from 1 to 3 or 1, 2 or 3, optionally h and/or h’ is at least 1;
IL is an integer ranging from 0 to 15, such as ranging from 1 to 15, ranging from 1 to 10, ranging from 1 to 5, ranging from 1 to 3, or 1, 2 or 3, optionally 11 is an integer ranging from 1 to 5 or 1, 2 or 3 with the proviso that at least one of h, h’ and 11 is at least 1, or a pharmaceutically acceptable salt, stereoisomer, solvate or polymorph thereof.
In some embodiments [MIFBM] is a moiety according to the chemical structure:
Figure imgf000006_0001
wherein XM is -(CH2)IM, -0-(CH2)IM, S-(CH2)IM, NRM-(CH2)IM, C(O)-(CH2)IM-, a polyethylene glycol (PEG) group containing from 1 to 8, such as 1-4 ethylene glycol residues or a -C(0)(CH2)IMNRM group;
RM is H or a C1-C3 alkyl group which is optionally substituted with one or two hydroxyl groups;
IM is 0-6, such as 1, 2, 3 or 4, such as 1.
In some embodiments, [IgGMB] is a group according to the chemical structure:
Figure imgf000007_0001
wherein DNP is a 2,4-dinitrophenyl group; or a group according to the chemical structure:
Figure imgf000007_0002
wherein Y’ is H or NO2;
X is O, CH2, NR1, S(O), S(O)2, -S(O)2O, -OS(O)2, or OS(O)2O; and
R1 is H, a C1-C3 alkyl group, or a -C(O)(Ci-C3) group; or a group according to the chemical structure:
Figure imgf000007_0003
wherein R1 is the same as above; and K” is 1-5 (such as 3-5, such as 4), or a group represented by the chemical formula:
Figure imgf000007_0004
wherein X’ is CH2, O, N-R1’, or S;
R1 is H or C1-C3 alkyl; and
Z is a bond, a monosaccharide, disaccharide, oligosaccharide, such as a sugar group selected from the monosaccharides, including aldoses and ketoses, and di saccharides, including those disaccharides described herein. Monosaccharide aldoses include monosaccharides such as aldotriose (D-glyceraldehdye, among others), aldotetroses (D- erythrose and D-Threose, among others), aldopentoses, (D-ribose, D-arabinose, D-xylose, D- lyxose, among others), aldohexoses (D-allose, D-altrose, D-Glucose, D-Mannose, D-gulose, D-idose, D-galactose and D-Talose, among others), and the monosaccharide ketoses include monosaccharides such as ketotriose (dihydroxyacetone, among others), ketotetrose (D- erythrulose, among others), ketopentose (D-ribulose and D-xylulose, among others), ketohexoses (D-Psicone, D-Fructose, D-Sorbose, D-Tagatose, among others), aminosugars, including galactoseamine , sialic acid, N-acetylglucosamine, among others and sulfosugars, including sulfoquinovose, among others. Exemplary disaccharides which find use in the present disclosure include sucrose (which may have the glucose optionally N-acetylated), lactose (which may have the galactose and/or the glucose optionally N-acetylated), maltose (which may have one or both of the glucose residues optionally N-acetylated), trehalose (which may have one or both of the glucose residues optionally N-acetylated), cellobiose (which may have one or both of the glucose residues optionally N-acetylated), kojibiose (which may have one or both of the glucose residues optionally N-acetylated), nigerose (which may have one or both of the glucose residues optionally N-acetylated), isomaltose (which may have one or both of the glucose residues optionally N-acetylated), P,P-trehalose (which may have one or both of the glucose residues optionally N-acetylated), sophorose (which may have one or both of the glucose residues optionally N-acetylated), laminaribiose (which may have one or both of the glucose residues optionally N-acetylated), gentiobiose (which may have one or both of the glucose residues optionally N-acetylated), turanose (which may have the glucose residue optionally N-acetylated), maltulose (which may have the glucose residue optionally N-acetylated), palatinose (which may have the glucose residue optionally N-acetylated), gentiobiluose (which may have the glucose residue optionally N- acetylated), mannobiose, melibiose (which may have the glucose residue and/or the galactose residue optionally N-acetylated), melibiulose (which may have the galactose residue optionally N-acetylated), rutinose, (which may have the glucose residue optionally N- acetylated), rutinulose and xylobiose, among others.
In some embodiments [IgGBM] is a group according to the chemical structure:
Figure imgf000008_0001
a-L-Rhamnose p-L-Rhamnose Phosphoryl Choline
Figure imgf000008_0002
Menadio
Figure imgf000008_0003
Carboxyethyl Lysine (R1 = Me) wherein XR is O, S NR1; and XM is O, NR1 or S, and
R1 is H or a C1-C3 alkyl group.
In some embodiments [IgGBM] is a group according to the chemical structure:
Figure imgf000009_0001
, wherein Xb is a bond, O, CH2 or NR1 or S; and
R1 is the same as above; or a group according to the chemical structure:
Figure imgf000009_0002
where RN02 is a dinitrophenyl group optionally linked through CH2, S(O), S(O)2, - S(O)2O, -OS(O)2, or OS(O)2O; or a dinitrophenyl group according to the chemical structure:
Figure imgf000009_0003
where K’” is 1-4 (such as 2-3, such as 3), or a a group according to a chemical structure which is set forth in FIGURE 67 hereof which is covalently attached to a [CON] group, a [LINKER] group or a [ASGPRBM] group through an amine group, such as a primary or secondary alkyl amine group which is optionally substituted on the amine group with a C1-C3 alkyl group.
In some embodiments [IgGBM] is a peptide according to the sequence (all references cited are incorporated by reference herein):
PAM (Fassina, et al., J. Mol. Recognit. 1996, 9, 564-569);
D-PAM (Verdoliva, et al., J. Immunol. Methods, 2002, 271, 77-88);
D-PAM-O (Dinon, et al. J. Mol. Recognit. 2011, 24, 1087-1094);
TWKTSRISIF (Krook, et al., J. Immunol. Methods 1998, 221, 151-157) SEQ ID
NO: 1;
FGRLVSSIRY (Krook, et al., J. Immunol. Methods 1998, 221, 151-157) SEQ ID NO:2;
Fc-III (DeLano,et al., Science 2000, 287, 1279-1283);
FcBP-1 (Kang, et al., J. Chromatogr. A 2016, 1466, 105-112);
FcBP-2 (Dias, et al., J. Am. Chem. Soc. 2006, 128, 2726-2732);
Fc-III-4c (Gong, et al., Bioconjug. Chem. 2016, 27, 1569-1573);
EPIHRSTLTALL (Ehrlich, et al., J. Biochem. Biophys. Method 2001, 49, 443-454)
SEQ ID NO:3;
APAR (Camperi, et al., Biotechnol. Lett. 2003, 25, 1545-1548) SEQ ID NO:4;
FcRM (Fc Receptor Mimetic, Verdoliva, et al., ChemBioChem 2005, 6, 1242-1253);
HWRGWV (Yang, et al., J. Peptide Res. 2006, 66, 110-137) SEQ ID NO:5;
HYFKFD (Yang, et al., J. Chromatogr. A 2009, 1216, 910-918) SEQ ID NO: 6;
HFRRHL (Menegatti, et al., J. Chromatogr. A 2016, 1445, 93-104) SEQ IDNO:7;
HWCitGWV (Menegatti, et al., J. Chromatogr. A 2016, 1445, 93-104) SEQ ID
NO:8;
D2AAG (Small Synthetic peptide ligand, Lund, et al., J. Chromatogr. A 2012, 1225, 158-167);
DAAG (Small Synthetic peptide ligand, Lund, et al., J. Chromatogr. A 2012, 1225, 158-167); cyclo[(N-Ac)S(A)-RWHYFK-Lact-E] (Menegatti, et al., 4 //a/. Chem. 2013, 85, 9229-9237)
(SEQ ID NO:9-Lact-E); cyclo[(N-Ac)-Dap(A)-RWHYFK-Lact-E] (Menegatti, et al., 4//a/. Chem. 2013, 85, 9229-9237) (SEQ ID NO: 10-Lact-E); cyclo[Link-M-WFRHYK] (Menegatti, et al., Biotechnol. Bioeng. 2013, 110, 857- SEQ ID NO: 11;
NKFRGKYK (Sugita, et al., Biochem. Eng. J. 2013, 79, 33-40) SEQ ID NO: 12;
NARKFYKG (Sugita, et al., Biochem. Eng. J. 2013, 79, 33-40) SEQ ID NO: 13;
FYWHCLDE (Zhao, et al., Biochem. Eng. J. 2014, 88, 1-11) SEQ ID NO: 14;
FYCHWALE (Zhao, et al., J. Chromatogr. A 2014, 1355, 107-114) SEQ ID NO: 15;
FYCHTIDE (Zhao, et al., J. Chromatogr. A 2014, 1359, 100-111) SEQ ID NO: 16;
Dual 1/3 (Zhao, et al., J. Chromatogr. A 2014, 1369, 64-72);
RRGW (Tsai, et al., Anal. Chem. 2014, 86, 2931-2938) SEQ ID NO: 17;
KHRFNKD (Yoo and Choi, BioChip J. 2015, 10, 88-94) SEQ ID NO: 18;
In some embodiments [ASGPRBM] is a group according to the chemical structure:
Figure imgf000011_0002
where X is 1-4 atoms in length and comprises O, S, N(RN1) or C(RN1)(RN1) groups such that when X is 1 atom in length, X is O, S, N(RN1) or C(RN1)(RN1), when X is 2 atoms in length, no more than 1 atom of X is O, S or N(RN1), when X is 3 or 4 atoms in length, no more than 2 atoms of X are O, S or N(RN1); where RN1 is H or a C1-C3 alkyl group optionally substituted with from 1-3 halo groups, such as F (in some embodiments, RN1 is H or methyl);
Ri and R3 are each independently H, -(CH2)KOH, -(CH2)KOCI-C4 alkyl, which is optionally substituted with from 1-3 halo (F, Cl, Br, or I) groups, C1-C4 alkyl, which is optionally substituted with from 1-3 halo (F, Cl, Br, or I) groups,
-(CH2)Kvinyl, O-(CH2)Kvinyl, -(CH2)Kalkynyl, -(CH2)KCOOH, -(CH2)KC(O)O-CI-C4 alkyl which is optionally substituted with from 1-3 halo, such as F groups, O-C(O)-Ci-C4 alkyl, which is optionally substituted with from 1-3 halo, such as F groups, -C(O)-Ci-C4 alkyl, which is optionally substituted with from 1-3 halo, such as F groups, or
Ri and R3 are each independently a
Figure imgf000011_0001
group, which is optionally substituted with up to three (such as 1) halo groups (such as F), C1-C4 alkyl groups, each of which is optionally substituted with from one to three halo groups, such as F, or one or two hydroxyl groups, or O-C1-C4 alkyl groups, each of which alkyl groups is optionally substituted with from one to three halo groups, such as F, or one or two hydroxyl groups, and
K is independently an integer ranging from 0-4 (such as 0, 1, 2, 3 or 4), or
Ri and R3 are each independently a group according to the chemical structure:
Figure imgf000012_0001
OH , where R7 is O-C1-C4 alkyl, which is optionally substituted with from 1 to 3 halo groups, such as F and 1 or 2 hydroxy groups, or R7 is a
-NR“R“ group
Figure imgf000012_0002
or Ri and R3 are each independently a group according to the structure:
Figure imgf000012_0003
wherein CYC is a ring selected from the group consisting of:
Figure imgf000012_0004
saturated carbocyclic, wherein each of LINKERX, Rc, and -(CH2)K- are attached to an open valence in CYC, including N-H;
Rc is absent, H, C1-C4 alkyl which is optionally substituted with from 1-3 halo (such as F) groups or 1-2 hydroxyl groups, or a group according to the structure:
Figure imgf000012_0005
where R4, Rs and Re are each independently, H, halo (F, Cl, Br, I), CN, NRN1RN2, -(CH2)KOH, -(CH2)KOCI-C4 alkyl, which is optionally substituted with from 1-3 halo (F, Cl, Br, I) groups, C1-C3 alkyl, which is optionally substituted with from 1-3 halo (F, Cl, Br, I) groups, -O-Ci-C3-alkyl, which is optionally substituted with from 1-3 halo, such as F groups, -(CH2)KC00H, -(CH2)KC(O)O-CI-C4 alkyl which is optionally substituted with from 1-3 halo, such as F groups, O-C(O)-Ci-C4 alkyl, which is optionally substituted with from 1-3 halo, such as F groups, -C(O)-Ci-C4 alkyl, which is optionally substituted with from 1-3 halo, such as F groups, or Rc is
Figure imgf000013_0001
where RN , RN1 and RN2 are each independently H or a C1-C3 alkyl group which is optionally substituted with from one to three halo groups, such as F, or one or two hydroxyl groups;
K is independently an integer ranging from 0-4 (0, 1, 2, 3 or 4);
K’ is an integer ranging from 1-4, such as 1;
RN3 is H, or a C1-C3 alkyl group which is optionally substituted with 1-3 halo groups, such as F or 1 or 2 hydroxy groups; and
RN4 is H, a C1-C3 alkyl group which is optionally substituted with 1-3 halo groups, such as F or 1 or 2 hydroxy groups, or RN4 is a
Figure imgf000013_0002
group, where K is sometimes 1;
LJNKERX is a linker group which is comprises to at least one [MIFBM/IgGBM] group and links the [MIFBM/IgGBM] group to the [ASGPRBM] through one or more optional [CON] groups, or
LiNKERX i
I - i is a linker group which contains at least one or more functional groups which can be used to covalently bond the linker group to at least one [MIFBM/IgGBM] group or optional [CON] group;
Figure imgf000013_0003
group where R and K are the same as above; RAM is H, a C1-C4 alkyl group optionally substituted with up to 3 halo groups (such as F) and one or two hydroxyl groups, a -(CH2)KCOOH group, a -(CH2)KC(O)O-CI-C4 alkyl group which is optionally substituted with from 1-3 halo, such as F groups, a
O-C(O)-Ci-C4 alkyl group, which is optionally substituted with from 1-3 halo, such as F groups, a -C(O)-Ci-C4 alkyl group, which is optionally substituted with from 1-3 halo, such as F groups, a -(CH2)K-NRN3RN4 group where RN3 is H, or a C1-C3 alkyl group which is optionally substituted with 1-3 halo groups, such as F or 1 or 2 hydroxy groups; and
RN4 is H, a C1-C3 alkyl group which is optionally substituted with 1-3 halo groups, such as F or 1 or 2 hydroxy groups, or RN4 is a
Figure imgf000014_0001
group (K is sometimes
1), or
Figure imgf000014_0002
group, where RTA is H, CN, NRN1RN2, -(CH2)KOH, -(CH2)KOCI-C4 alkyl, which is optionally substituted with from 1-3 halo (F, Cl, Br, or I) groups, C1-C4 alkyl, which is optionally substituted with from 1-3 halo (F, Cl, Br, or I) groups, -(CH2)KCOOH, -(CH2)KC(O)O-CI-C4 alkyl which is optionally substituted with from 1-3 halo, such as F groups, O-C(O)-Ci-C4 alkyl, which is optionally substituted with from 1-3 halo, such as F groups, -C(O)-Ci-C4 alkyl, which is optionally substituted with from 1-3 halo, such as F groups, or RTA is a C3-C10 aryl or a three- to ten-membered heteroaryl group containing up to 5 heteroaryl atoms, each of said aryl or heteroaryl groups being optionally substituted with up to three (such as 1) CN, NRNiRN2, -(CH2)KOH, -(CH2)KOCI-C4 alkyl, which is optionally substituted with from 1-3 halo (F, Cl, Br, or I) groups, C1-C3 alkyl, which is optionally substituted with from 1-3 halo (F, Cl, Br, or I) groups or 1 or 2 hydroxy groups, -O-Ci-C3-alkyl, which is optionally substituted with from 1-3 halo, such as F groups, -(CH2)KCOOH, -(CH2)KC(O)O-CI-C4 alkyl which is optionally substituted with from 1-3 halo, such as F groups, O-C(O)-Ci-C4 alkyl, which is optionally substituted with from 1-3 halo, such as F groups or -(CH2)KC(O)-CI-C4 alkyl which is optionally substituted with from 1-3 halo, such as F groups, or
RTA is
Figure imgf000014_0003
or
RTA is a
Figure imgf000015_0001
group which is optionally substituted with up to three, such as
1 C1-C3 alkyl groups which are optionally substituted with up to three halo (such as F) groups,
Figure imgf000015_0002
group, wherein RN , RN1 and RN2 are each independently H or a C1-C3 alkyl group which is optionally substituted with from one to three halo groups, such as F, or one or two hydroxyl groups and wherein each -(CH2)K group is optionally substituted with 1-4, such as 1 or 2, C1-C3 alkyl groups which are optionally substituted with from 1-3 fluoro groups or 1-2 hydroxyl groups; and K is independently 0-4 (0, 1, 2, 3 or 4).
In some embodiments, [CON] is a connector moiety (including a [MULTICON]) as otherwise described herein.
In some embodiments, [LINKER] is a linking moiety as otherwise described herein which links [MIFBM/IgGBM] to the [ASGPRBM] group and optionally contains one or more connector moieties (which optionally connect(s) more than one chemical moiety to provide said linking moiety or which connects said linking moiety to said [MIFBM/IgGBM] group or said [ASGPRBM] group, or a pharmaceutically acceptable salt, stereoisomer, solvate or polymorph thereof.
In some embodiments, X is -O-C(RN1)(RN1),
C(RN1)(RN1)-O-, -S-C(RN1)(RN1), C(RN1)(RN1)-S-, N(RN1)-C(RN1)(RN1), C(RN1)(RN1)-N(RN1) or C(RN1)(RN1)-C(RN1)(RN1) when X is 2 atoms in length,
X is -O-C(RN1)(RN1)-C(RN1)(RN1), C(RN1)(RN1)-O-C(RN1)(RN1)-, -O-C(RN1)(RN1)-O-, -O-C(RN1)(RN1)-S-, -O-C(RN1)(RN1)-N(RN1)-, -S-C(RN1)(RN1)-C(RN1)(RN1), C(RN1)(RN1)-S-C(RN1)(RN1)-, C(RN1)(RN1)-C(RN1)(RN1)-S, -S-C(RN1)(RN1)-S-, -S-C(RN1)(RN1)-O-, -S-C(RN1)(RN1)-N(RN1)-, N(RN1)-C(RN1)(RN1)-C(RN1)(RN1), C(RN1)(RN1)-N(RN1)-C(RN1)(RN1), C(RN1)(RN1)-C(RN1)(RN1)- N(RN1), N(RN1)-C(RN1)(RN1)-N(RN1) or C(RN1)(RN1)-C(RN1)(RN1)- C(RN1)(RN1) when X is 3 atoms in length, and X is -O-C(RN1)(RN1)-C(RN1)(RN1)-C(RN1)(RN1), C(RN1)(RN1)-O-C(RN1)(RN1)- (RN1)(RN1)-, -O-C(RN1)(RN1)-O-C(RN1)(RN1)-, -S-C(RN1)(RN1)-C(RN1)(RN1)- C(RN1)(RN1)-, C(RN1)(RN1)-S-C(RN1)(RN1)-C(RN1)(RN1)-, C(RN1)(RN1)-(RN1)(RN1)- S-C(RN1)(RN1)-, -S-C(RN1)(RN1)-S-C(RN1)(RN1)-, N(RN1)-C(RN1)(RN1)- C(RN1)(RN1)- C(RN1)(RN1)-, C(RN1)(RN1)-N(RN1)-C(RN1)(RN1)-C(RN1)(RN1), C(RN1)(RN1)-C(RN1)(RN1)- N(RN1), N(RN1)-C(RN1)(RN1)-N(RN1) or C(RN1)(RN1)- C(RN1)(RN1)- C(RN1)(RN1) when X is 4 atoms in length where RN1 is the same as above. In some embodiments, RN1 is H.
In embodiments, X is OCH2 or CH2O. In some embodiments RN1 is H.
In embodiments, the [ASGPRBM] group is a group according to the chemical structure:
Figure imgf000016_0001
where Ri, R2 and R3 are the same as above, or a pharmaceutically acceptable salt, stereoisomer, solvate or polymorph thereof.
In some embodiments, the [ASGPRBM] group is a group according to the chemical structure:
Figure imgf000016_0002
where RA is a C1-C3 alkyl group which is optionally substituted with 1-5 halo (such as F) groups (In some embodiments, RA is a methyl or ethyl group which is optionally substituted with from 1-3 fluoro groups);
ZA is -(CH2)IM, -O-(CH2)IM, S-(CH2)IM, NRM-(CH2)IM, C(O)-(CH2)IM-, a PEG group containing from 1 to 8, such as 1-4 ethylene glycol residues or a -C(0)(CH2)IMNRM group (In some embodiments, a PEG containing group comprising from 1 to 8 ethylene glycol, such as 2-4 ethylene glycol residues) where IM and RM are the same as above; and
ZB is absent, (CH2)IM, C(0)-(CH2)IM- or C(0)-(CH2)IM-NRM, where IM and RM are the same as above.
In some embodiments, Ri and R3 are each independently often a group according to the chemical structure:
Figure imgf000017_0001
are the same as above.
In some embodiments, in the above-described compounds,
Figure imgf000017_0002
In some embodiments, the compounds include the compounds which are presented in
FIGURES 1, 7 and 13. In some embodiments, additional compounds are presented in FIGURES 16-66 and include final compounds set forth therein and intermediates which are used to make final compounds pursuant to the present disclosure.
In some embodiments, Ri and R3 of the [ASGPRBM] group include those moieties which are presented in FIGURE 68 hereof. In some embodiments, R2 of the [ASGPRBM] group include those moieties which are presented in FIGURE 69 hereof.
In some embodiments, the IgGBM group is a FCIII group according to the chemical
Figure imgf000018_0002
or a FcIII-4c peptide represented
Figure imgf000018_0001
. (SEQ ID NO:20).
In some aspects, the present disclosure is directed to a pharmaceutical composition comprising an effective amount of a compound according to the present disclosure in combination with a pharmaceutically acceptable carrier, additive or excipient, optionally in combination with at least one additional bioactive agent.
In some aspects, the present disclosure is directed to a method of treating a disease state or condition where MIF is related to or contributes to a disease state and or condiiton or the symptomology associated with the disease state or condition. These disease states and/or conditions include, autoimmune diseases and numerous inflammatory diseases for example, rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), Alzheimer’s disease, atherosclerosis, heart disease, stroke and cancer (including leukemia), among numerous others. In some embodiments, the method of treatment comprises administering to a patient or subject in need of therapy an effective amount of at least one compound according to the present disclosure, optionally in combination with an additional bioactive agent to reduce the likelihood of, inhibit and/or treat the disease state or condition by removing MIF associated with the disease state and/or condition from the circulation of the patient or subject. In experiments, bi-functional molecules of the present disclosure, MIF-GN3 were able to induce the degradation of human MIF injected in mice. Additional work focuses on the evaluation of MIF-GN3 in autoimmune disease models as well as seeking to optimize the bifunctional MIF degrader using different MIF and ASGPr ligands. IgG-targeting molecules were able to recapitulate our results with MIF in cell culture and mediated the removal of IgG from mouse serum. Thus, the present disclosure provides a useful platform for the development of new therapeutics for these critical diseases.
BRIEF DESCRIPTION OF THE FIGURES
FIGURE 1 shows representative compounds according to some embodiments. Note that the figure discloses compound 3w (negative control for MIF inhibition), MIF-NVS- PEGnGN3, MIFGN3, MIF-PEGnGN3, MIF-AcF3-l, MIF-AcF3-2 and MIF-AcF3-3. Note that n in the PEG linker sometimes ranges from 1-12, such as 1 to 10, 2 to 8, 2 to 6, 2 to 5 or 1, 2, 3 or 4.
FIGURE 2 shows fluorescence polarization data of MIF-FITC binding to human MIF, indicating that our MIF -binding moiety binds MIF, in accordance with some embodiments. Bifunctional molecules WJ-PEG4-GN3, WJ-PEG2-GN3, and NVS-PEG3- GN3 bound competitively with MIF-FITC, indicating that the bifunctional molecules maintain the ability to bind human MIF.
FIGURE 3 shows that bifunctional molecules are able to deplete human MIF from the supernatant of culture HepG2 cells, in accordance with some embodiments.
FIGURE 4 shows that MIF internalized by HepG2 cells is trafficked to lysosomes, in accordance with some embodiments.
FIGURE 5 shows that MIF-GN3 mediates the depletion of injected human MIF from mice, in accordance with some embodiments.
FIGURE 6 shows that MIF-GN3 is able to delay tumor growth in a mouse model of prostate cancer, in accordance with some embodiments.
FIGURE 7 shows molecules DNP-GN3 and DNP-AcF3-3, which are bifunctional moelcules that bind to anti-DNP IgG and ASGPR, in accordance with some embodiments.
FIGURE 8 shows that DNP-GN3 and DNP-AcF3-3 mediate the formation of a ternary complex between HepG2 cells and anti-DNP, in accordance with some embodiments.
FIGURE 9 shows that DNP-GN3 and DNP-AcF3-3 mediate the uptake of alexa 488- labeled anti-DNP by HepG2 cells, in accordance with some embodiments.
FIGURE 10 shows that DNP-GN3 and DNP-AcF3-3 mediate the localization of alexa 568 labeled anti-DNP to late endosomes and lysosomes, in accordance with some embodiments.
FIGURE 11 shows that DNP-AcF3-3 mediates the degradation of alexa 488-labeled anti-DNP in HepG2 cells, in accordance with some embodiments.
FIGURE 12 shows that DNP-GN3 mediates the depletion of anti-DNP from mouse serum, in accordance with some embodiments.
FIGURE 13 shows the structures of IgG-degrading molecules IBA-GN3, Triazine- GN3, FcIII-GN3, and FcIII-4c-GN3, in accordance with some embodiments.
FIGURE 14 shows that FcIII-GN3 mediates the uptake of human IgG into HepG2 cells, in accordance with some embodiments. Experiment performed as described above.
FIGURE 15 shows that FcIII-GN3 mediates the localization of IgG to late endosomes in HepG2 cells, in accordance with some embodiments. Experiment performed as described above.
FIGURES 16-18 show the synthesis of PEG linkers used in several molecules , in accordance with some embodiments.
FIGURES 19-21 show the synthesis of ASGPR-binding precursors and ligands used in several molecules , in accordance with some embodiments.
FIGURES 22-26 show the synthesis of valency linkers used in several molecules , in accordance with some embodiments.
FIGURES 27-28 show the synthesis of MIF ligands used in several bifunctional molecules, in accordance with some embodiments.
FIGURE 29 describes the synthesis of the bifunctional molecule MIF-NVS-PEGn- GN3, in accordance with some embodiments.
FIGURE 30 describes the synthesis of bifunctional molecules MIF-GN3 and MIF- PEGn-GN3, in accordance with some embodiments.
FIGURE 31 describes the synthesis of the bifunctional molecule targeting MIF and ASGPR, containing one bicyclic ASGPR AcF3 ligands, in accordance with some embodiments.
FIGURE 32 describes the synthesis of the bifunctional molecule targeting MIF and ASGPR, containing two bicyclic ASGPr AcF3 ligands, in accordance with some embodiments.
FIGURE 33 describes the synthesis of the bifunctional molecule targeting MIF and ASGPR, containing three bicyclic ASGPr ligands, in accordance with some embodiments.
FIGURE 34 shows the synthesis of DNP-GN3, in accordance with some embodiments.
FIGURE 35 shows the synthesis of DNP-AcF3-3, in accordance with some embodiments.
FIGURE 36 shows the synthetic scheme used to obtain IBA-GN3, in accordance with some embodiments.
FIGURE 37 shows the synthesis of triazine-GN3, in accordance with some embodiments.
FIGURE 38 shows the synthetic scheme used to access FcIII-GN3, in accordance with some embodiments.
FIGURE 39 shows the synthetic scheme used to access FcIII-4c-GN3, in accordance with some embodiments.
FIGURES 40-43 describe the synthesis of bifunctional molecules targeting MIF and ASGPr, containing three bicyclic ASGPR ligands with different substitutions on the 2-amine of the sugar, in accordance with some embodiments.
FIGURE 44 shows the synthesis of compound MIF- 18-3, in accordance with some embodiments.
FIGURE 45 shows the synthesis of compound MIF-31-3, in accordance with some embodiments.
FIGURE 46 shows the synthesis of compound MIF-15-3, in accordance with some embodiments.
FIGURE 47 shows the synthesis of compound MIF- 19-3, in accordance with some embodiments.
FIGURE 48 shows the synthesis of compound MIF- 16-3, in accordance with some embodiments.
FIGURE 49 shows the synthesis of compound MIF-20-3, in accordance with some embodiments.
FIGURE 50 shows the synthesis of compound MIF- 14-3, in accordance with some embodiments.
FIGURE 51 shows the synthesis of compound MIF-21-3, in accordance with some embodiments.
FIGURES 52-66 show the synthesis of a number of MIF-binding compounds with various ASGPRBM moieties, in accordance with some embodiments.
FIGURE 67 shows exemplary IgGBM groups each of which is covalently attached to a [CON] group, a [LINKER] group or a [ASGPRBM] group through an amine group, such as a primary or secondary alkyl amine group which is optionally substituted on the amine group with a C1-C3 alkyl group, in accordance with some embodiments.
FIGURE 68 shows exemplary Ri and R3 substituents on ASGPRBM groups, in accordance with some embodiments.
FIGURE 69 shows exemplary R2 substituents on ASGPRBM groups, in accordance with some embodiments.
DETAILED DESCRIPTION
In some aspects, the present disclosure provides bi-functional molecules that utilize the endo-lysosomal machinery in the liver to eliminate MIF and/or IgG. In some embodiments, the present disclosure utilizes bi-functional molecules to target MIF or IgG and is modular and versatile. This offers convenience to apply medicinal chemistry to optimize the molecules. For example, in preliminary experiments, incorporation of more optimized ASGPr binding motifs have been shown to enhance MIF or IgG degradation in vitro. Since the bi-functional molecule is capable of degrading MIF or IgG, it does not require the MIF- binding motif (MIFBM) or the IgG binding moiety (IgGBM)to be an inhibitor, but in practice many of these binders will function as inhibitors of MIF or IgG. The reality of this dual inhibitory action opens up opportunity for the development of potent MIF or IgG binders, which might have been overlooked before for not being able to inhibit various PPIs. Compared to monoclonal antibodies, the small molecule approach of the present disclosure enables simpler, cheaper and more consistent manufacturing practices and provides for a therapeutic approach with fewer potential immugenic side effects.
In some aspects, the present disclosure is directed to bi-functional compounds which find use as pharmaceutical agents in the treatment of disease states and/or conditions which are mediated through macrophage migration inhibitory factor (MIF) or Immunoglobulin G (IgG). In some aspects, the present disclosure is directed to pharmaceutical compositions which comprise these bi-functional compounds as well as methods for treating disease states and/or conditions which are mediated through MIF or IgG or where MIF and IgG are contributing factors to the development and perpetuation of diseases and/or conditions, especially including autoimmune diseases and cancer, among others. In some aspects, present disclosure provides a molecular strategy to lower plasma MIF level or IgG levels in patients with autoimmune diseases or certain types of cancers and other diseases. The bi- functional molecule construct is comprised of a MIF-targeting motif that is derived from small molecule MIF ligands or an IgG-binding motif which binds to IgG, and an ASGPr- targeting motif that binds to hepatocyte asialoglycoprotein receptor (ASGPr). The compounds selectively bind MIF or IgG in plasma and subsequently engage the endo- lysosomal pathway of hepatocytes through ASGPr. As a consequence, MIF or IgG is internalized and degraded by hepatocytes, thus resulting in potential attenuation of corresponding disease symptoms which are modulated through MIF and/or IgG.
In accordance with some embodiments, conventional chemical synthetic and pharmaceutical formulation methods, as well as pharmacology, molecular biology, microbiology, and recombinant DNA techniques within the skill of the art are employed. Such techniques are well-known and are otherwise explained fully in the literature.
Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise (such as in the case of a group containing a number of carbon atoms), between the upper and lower limit of that range and any other stated or intervening value in that stated range is encompassed within the disclosure. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges is also encompassed within the disclosure, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either both of those included limits are also included in the disclosure.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present disclosure, some exemplary methods and materials are now described.
It is to be noted that as used herein and in the appended claims, the singular forms "a," “an”, "and" and "the" include plural references unless the context clearly dictates otherwise.
Furthermore, the following terms shall have the definitions set out below. It is understood that in the event a specific term is not defined hereinbelow, that term shall have a meaning within its typical use within context by those of ordinary skill in the art.
Definition
The term “compound”, as used herein, unless otherwise indicated, refers to any specific chemical compound disclosed herein and includes tautomers, regioisomers, geometric isomers, stereoisomers and where applicable, optical isomers (enantiomers) thereof, as well as pharmaceutically acceptable salts and derivatives (including prodrug forms) thereof. Within its use in context, the term compound generally refers to a single compound, but also may include other compounds such as stereoisomers, regioisomers and/or optical isomers (including racemic mixtures) as well as specific enantiomers or enantiomerically enriched mixtures of disclosed compounds. The term also refers, within context, to prodrug forms of compounds which have been modified to facilitate the administration and delivery of compounds to a site of activity. It is noted that in describing the present compounds, numerous substituents, linkers and connector molecules and variables associated with same, among others, are described. The use of a bond presented as - signifies that a single bond is present or absent, depending on the context of the chemistry described, including the attachment of the bond to another moiety. The use of a bond presented as - signifies that a single bond or a double bond is intended depending on the context of the chemistry described. It is understood by those of ordinary skill that molecules which are described herein are stable compounds as generally described hereunder.
The term “patient” or “subject” is used throughout the specification within context to describe an animal, generally a mammal, such as a human, to whom treatment, including prophylactic treatment (prophylaxis, including especially as that term is used with respect to reducing the likelihood of metastasis of an existing cancer), with the compositions according to the present disclosure is provided. For treatment of those infections, conditions or disease states which are specific for a specific animal such as a human patient or a patient of a particular gender, such as a human male or female patient, the term patient refers to that specific animal. Compounds according to the present disclosure are useful for the treatment of numerous disease states including autoimmune diseease states and/or conditions and inflammatory disease states and/or conditions as well as cancer, including especially for use in reducing the likelihood of metastasis or recurrence of a cancer.
The term “effective” is used herein, unless otherwise indicated, to describe an amount of a compound or composition which, in context, is used to produce or effect an intended result, whether that result relates to the inhibition of the effects of a disease state (e.g. an autoimmune disease such as rheumatoid arthritis (RA) or systemic lupus erythematosus (SLE), among others, atherosclerosis, heart disease or stroke, among numerous others or a cancer, including leukemia) on a subject or the treatment or prophylaxis of a subject for secondary conditions, disease states or manifestations of disease states as otherwise described herein. This term subsumes all other effective amount or effective concentration terms (including the term “therapeutically effective”) which are otherwise described in the present application. The terms “treat”, “treating”, and “treatment”, etc., as used herein, refer to any action providing a benefit to a patient at risk for a disease state or condition for which a MIF protein may be removed, such as an autoimmune disease including rheumatoid arthritis (RA) or systemic lupus erythematosus (SLE), among others, atherosclerosis, heart disease, stroke and cancer (including leukemia) including recurrence and/or metastasis of cancer, improvement in the condition through lessening or suppression of at least one symptom of the disease state or condition, inhibition of one or more manifestations of the disease state (e.g., plaque formation, heart disease, cancer growth, reduction in cancer cells or tissue), prevention, reduction in the likelihood or delay in progression of a disease state or condition or manifestation of the disease state or condition, especially including plaque formation in atheroslerosis, deterioration of tissue and inflammation in rheumatoid arthritis, further damage to cardiovascular tissue in heart disease, further damage to central nervous tissue in stroke, cancer, its recurrence or metastasis of the cancer, prevention or delay in the onset of disease states or conditions which occur secondary to the disease state or condition including cancer recurrence or metastasis, among others. Treatment, as used herein, encompasses both prophylactic and therapeutic treatment, depending on the context of the treatment. The term “prophylactic” when used, means to reduce the likelihood of an occurrence or the severity of an occurrence within the context of treatment of disease state or condition, as otherwise described hereinabove.
As used in this application, the terms “about” and “approximately” are used as equivalents. Any numerals used in this application with or without about/approximately are meant to cover any normal fluctuations appreciated by one of ordinary skill in the relevant.
The term “macrophage migration inhibitory factor binding moiety” or “MIFBM” refers to a chemical moiety on one end of the bifunctional compounds according to the present disclosure which is capable of binding to a circulating MIF protein which is associated with or contributes to a disease state or condition as otherwise described herein. In the present disclosure, the MIFBM is capable of binding to the circulating MIF protein, forming a complex with the present compounds, and delivering the bound protein to a hepatocyte whereupon the other end of the bifunctional molecule which contains an asialoglycoprotein receptor binding moiety (ASGPRBM) and can bind to the surface of a hepatocyte, respectively. Once attached to the hepatocyte, the bifunctional molecule to which is bound circulating protein is internalized by the hepatocyte through an endocytosis mechanism whereupon the cell will destroy the protein via a lysosomal degradation or other degradation pathway. The term “immunoglobulin G binding moiety” or “IgGBM” is used to describe a moiety which binds to circulating IgG immunoglobulin, forming a complex with bifunctional molecules according to the present disclosure to be ultimately destroyed in hepatocytes. In certain instances in describing the present disclosure, the terms MIFBM and IgGBM are used synonymously.
Exemplary MIFBMs for inclusion in bifunctional compounds according to the present disclosure include moieties found in bifunctional chemical structures which appear in Figure 1, attached hereto. In some embodiments, MIFBMs include moieties according to the
Figure imgf000026_0001
wherein XM is -(CH2)IM, -0-(CH2)IM, S-(CH2)IM, NRM-(CH2)IM, C(O)-(CH2)IM-, a PEG group containing from 1 to 8, such as 1-4 ethylene glycol residues or a -C(O)(CH2)IMNRM group;
RM is H or a C1-C3 alkyl group which is optionally substituted with one or two hydroxyl groups; and
IM is 0-6, such as 1, 2, 3 or 4.
The term “asialoglycoprotein receptor binding moiety” (“ASGPRBM”) refers to a binding moiety which binds to hepatocyte asialoglycoprotein receptor. This binding moiety is also a component of the presently claimed bifunctional compounds bound to the MIHBM moiety through a linker. The ASGPRBM selectively binds to hepatocyte asialoglycoprotein receptor on the surface of hepatocytes. It is through this moiety that bifunctional compounds complexed with circulating protein bind to hepatocytes. Once bound to the hepatocyte, the circulating MIF protein is taken into the hepatocytes via a phagocytosis mechanism wherein the circulating protein is degraded through lysosomal degradation.
Exemplary ASGPRBM groups for use in compounds according to the present disclosure, among others, include moieties according to the chemical structures:
Figure imgf000027_0001
where X is 1-4 atoms in length and comprises O, S, N(RN1) or C(RN1)(RN1) groups such that when X is 1 atom in length, X is O, S, N(RN1) or C(RN1)(RN1), when X is 2 atoms in length, no more than 1 atom of X is O, S or N(RN1), when X is 3 or 4 atoms in length, no more than 2 atoms of X are O, S or N(RN1); where RN1 is H or a C1-C3 alkyl group optionally substituted with from 1-3 halo groups, such as F (in some embodiments, RN1 is H or methyl); R1 and R3 are each independently H, -(CH2)KOH, -(CH2)KOC1-C4 alkyl, which is optionally substituted with from 1-3 halo (F, Cl, Br, or I) groups, C1-C4 alkyl, which is optionally substituted with from 1-3 halo (F, Cl, Br, or I) groups, -(CH2)Kvinyl, O-(CH2)Kvinyl, -(CH2)Kalkynyl, -(CH2)KCOOH, -(CH2)KC(O)O-C1-C4 alkyl which is optionally substituted with from 1-3 halo, such as F groups, O-C(O)-C1-C4 alkyl, which is optionally substituted with from 1-3 halo, such as F groups, -C(O)-C1-C4 alkyl, which is optionally substituted with from 1-3 halo, such as F groups, or
Figure imgf000027_0002
group, which is optionally substituted with up to three (such as 1) halo groups (such as F), C1-C4 alkyl groups, each of which is optionally substituted with from one to three halo groups, such as F, or one or two hydroxyl groups, or O-C1-C4 alkyl groups, each of which alkyl groups is optionally substituted with from one to three halo groups, such as F, or one or two hydroxyl groups, and K is independently an integer ranging from 0 to 4 (0, 1, 2, 3 or 4), or R1 and R3 are each independently a group according to the chemical structure:
Figure imgf000027_0003
, where R7 is O-C1-C4 alkyl, which is optionally substituted with from 1 to 3 halo groups, such as F and 1 or 2 hydroxy groups, or R7 is a -NRN3RN4 group
Figure imgf000027_0004
or R1 and R3 are each independently a group according to the structure:
Figure imgf000028_0001
wherein CYC is a ring selected from the group consisting of: 5
Figure imgf000028_0002
saturated carbocyclic, wherein each of LINKERX, RC, and –(CH2)K- are attached to an open valence in CYC, including N-H; RC is absent, H, C1-C4 alkyl which is optionally substituted with from 1-3 halo (such 10 as fluoro) groups or 1-2 hydroxyl groups, or a group according to the structure:
Figure imgf000028_0003
where R4, R5 and R6 are each independently, H, halo (F, Cl, Br, I), CN, NRN1RN2, -(CH2)KOH, -(CH2)KOC1-C4 alkyl, which is optionally substituted with from 1-3 halo (F, Cl, Br, or I) groups, C1-C3 alkyl, which is optionally substituted with from 1-3 halo (F, Cl, 15 Br, or I) groups, -O-C1-C3-alkyl, which is optionally substituted with from 1-3 halo, such as F groups, -(CH2)KCOOH, -(CH2)KC(O)O-C1-C4 alkyl which is optionally substituted with from 1-3 halo, such as F groups, O-C(O)-C1-C4 alkyl, which is optionally substituted with from 1-3 halo, such as F groups, -C(O)-C1-C4 alkyl, which is optionally substituted with from 1-3 halo, such as F groups, or 20 RC is
Figure imgf000029_0001
where RN, RN1 and RN2 are each independently H or a C1-C3 alkyl group which is optionally substituted with from one to three halo groups, such as F, or one or two hydroxyl groups; K is independently an integer ranging from 0 to 4 (0, 1, 2, 3 or 4); K’ is an integer ranging from 1-4, such as 1; RN3 is H, or a C1-C3 alkyl group which is optionally substituted with 1-3 halo groups, such as F or 1 or 2 hydroxy groups; and RN4 is H, a C1-C3 alkyl group which is optionally substituted with 1-3 halo groups, such as F or 1 or 2 hydroxy groups, or RN4 is a
Figure imgf000029_0002
group, where K is sometimes 1; is a linker group which is comprises to at least one [MIFBM/IgGBM] group and links the [MIFBM/IgGBM] group to the [ASGPRBM] through one or more optional [CON] groups, or is a linker group which contains at least one or more functional groups which can be used to covalently bond the linker group to at least one [MIFBM/IgGBM] group or optional [CON] group;
Figure imgf000029_0003
group where RN1 and K are the same as above; RAM is H, a C1-C4 alkyl group optionally substituted with up to 3 halo groups (such as F) and one or two hydroxyl groups, a -(CH2)KCOOH group, a -(CH2)KC(O)O-C1-C4 alkyl group which is optionally substituted with from 1-3 halo, such as F groups, a O-C(O)-C1-C4 alkyl group, which is optionally substituted with from 1-3 halo, such as F groups, a -C(O)-C1-C4 alkyl group, which is optionally substituted with from 1-3 halo, such as F groups, a -(CH2)K-NRN3RN4 group where RN3 is H, or a C1-C3 alkyl group which is optionally substituted with 1-3 halo groups, such as F or 1 or 2 hydroxy groups; and RN4 is H, a C1-C3 alkyl group which is optionally substituted with 1-3 halo groups, such as F or 1 or 2 hydroxy groups, or RN4 is a
Figure imgf000030_0001
group (K is sometimes 1), or
Figure imgf000030_0002
group, where RTA is H, CN, NRN1RN2, -(CH2)KOH, -(CH2)KOC1-C4 alkyl, which is optionally substituted with from 1-3 halo (F, Cl, Br, or I) groups, C1-C4 alkyl, which is optionally substituted with from 1-3 halo (F, Cl, Br, or I) groups, -(CH2)KCOOH, -(CH2)KC(O)O-C1-C4 alkyl which is optionally substituted with from 1-3 halo, such as F groups, O-C(O)-C1-C4 alkyl, which is optionally substituted with from 1-3 halo, such as F groups, -C(O)-C1-C4 alkyl, which is optionally substituted with from 1-3 halo, such as F groups, or RTA is a C3-C10 aryl or a three- to ten-membered heteroaryl group containing up to 5 heteroaryl atoms, each of said aryl or heteroaryl groups being optionally substituted with up to three (such as 1) CN, NRN1RN2, -(CH2)KOH, -(CH2)KOC1-C4 alkyl, which is optionally substituted with from 1-3 halo (F, Cl, Br, or I) groups, C1-C3 alkyl, which is optionally substituted with from 1-3 halo (F, Cl, Br, or I) groups or 1 or 2 hydroxy groups, -O-C1-C3-alkyl, which is optionally substituted with from 1-3 halo, such as F groups, -(CH2)KCOOH, -(CH2)KC(O)O-C1-C4 alkyl which is optionally substituted with from 1-3 halo, such as F groups, O-C(O)-C1-C4 alkyl, which is optionally substituted with from 1-3 halo, such as F groups or -(CH2)KC(O)-C1-C4 alkyl which is optionally substituted with from 1-3 halo, such as F groups, or RTA is
Figure imgf000030_0003
(in some embodiments, RTA is a
Figure imgf000030_0004
group which is optionally substituted with up to three, such as 1 C1-C3 alkyl groups which are optionally substituted with up to three halo (such as F) groups,
Figure imgf000030_0005
group,) wherein RN, RN1 and RN2 are each independently H or a C1-C3 alkyl group which is optionally substituted with from one to three halo groups, such as F, or one or two hydroxyl groups and wherein each -(CH2)K group is optionally substituted with 1-4, such as 1 or 2, C1-C3 alkyl groups which are optionally substituted with from 1-3 fluoro groups or 1-2 hydroxyl groups; and K is independently 0-4 (0, 1, 2, 3 or 4).
In some embodiments, [CON] is a connector moiety (including a [MULTICON]) as otherwise described herein.
In some embodiments, [LINKER] is a linking moiety as otherwise described herein which links [MIFBM/IgGBM] to the [ASGPRBM] group and optionally contains one or more connector moieties (which optionally connect(s) more than one chemical moiety to provide said linking moiety or which connects said linking moiety to said [MIFBM/IgGBM] group or said [ASGPRBM] group, or a pharmaceutically acceptable salt, stereoisomer, solvate or polymorph thereof.
In some embodiments, X is -O-C(RN1)(RN1),
C(RN1)(RN1)-O-, -S-C(RN1)(RN1), C(RN1)(RN1)-S-, N(RN1)-C(RN1)(RN1), C(RN1)(RN1)-N(RN1) or C(RN1)(RN1)-C(RN1)(RN1) when X is 2 atoms in length,
X is -O-C(RN1)(RN1)-C(RN1)(RN1), C(RN1)(RN1)-O-C(RN1)(RN1)-, -O-C(RN1)(RN1)-O-, -O-C(RN1)(RN1)-S-, -O-C(RN1)(RN1)-N(RN1)-, -S-C(RN1)(RN1)-C(RN1)(RN1), C(RN1)(RN1)-S-C(RN1)(RN1)-, C(RN1)(RN1)-C(RN1)(RN1)-S, -S-C(RN1)(RN1)-S-, -S-C(RN1)(RN1)-O-, -S-C(RN1)(RN1)-N(RN1)-, N(RN1)-C(RN1)(RN1)-C(RN1)(RN1), C(RN1)(RN1)-N(RN1)-C(RN1)(RN1), C(RN1)(RN1)-C(RN1)(RN1)- N(RN1), N(RN1)-C(RN1)(RN1)-N(RN1) or C(RN1)(RN1)-C(RN1)(RN1)- C(RN1)(RN1) when X is 3 atoms in length, and
X is -O-C(RN1)(RN1)-C(RN1)(RN1)-C(RN1)(RN1), C(RN1)(RN1)-O-C(RN1)(RN1)- (RN1)(RN1)_, -O-C(RN1)(RN1)-O-C(RN1)(RN1)-, -S-C(RN1)(RN1)-C(RN1)(RN1)- C(RN1)(RN1)-, C(RN1)(RN1)-S-C(RN1)(RN1)-C(RN1)(RN1)-, C(RN1)(RN1)-(RN1)(RN1)- S-C(RN1)(RN1)-, -S-C(RN1)(RN1)-S-C(RN1)(RN1)-, N(RN1)-C(RN1)(RN1)- C(RN1)(RN1)- C(RN1)(RN1)-, C(RN1)(RN1)-N(RN1)-C(RN1)(RN1)-C(RN1)(RN1), C(RN1)(RN1)-C(RN1)(RN1)- N(RN1), N(RN1)-C(RN1)(RN1)-N(RN1) or C(RN1)(RN1)- C(RN1)(RN1)- C(RN1)(RN1) when X is 4 atoms in length where RN1 is the same as above. Most often, RN1 is H.
In some embodiments, X is OCH2 or CH2O. In some embodiments, RN1 is H.
In embodiments, the [ASGPRBM] group is a group according to the chemical structure:
Figure imgf000032_0001
where R1, R2 and R3 are the same as above, or a pharmaceutically acceptable salt, stereoisomer, solvate or polymorph thereof. In embodiments, the [ASGPRBM] group is a group according to the chemical structure:
Figure imgf000032_0002
where RA is a C1-C3 alkyl group which is optionally substituted with 1-5 halo (such as fluoro) groups (in some embodiments, RA is a methyl or ethyl group which is optionally substituted with from 1-3 fluoro groups); ZA is -(CH2)IM, -O-(CH2)IM, S-(CH2)IM, NRM-(CH2)IM, C(O)-(CH2)IM-, a polyethylene glycol (PEG) group containing from 1 to 8, such as 1-4 ethylene glycol residues or a - C(O)(CH2)IMNRM group (such as a PEG containing group comprising from 1 to 8 ethylene glycol, such as 2-4 ethylene glycol residues) where IM and RM are the same as above; and ZB is absent, (CH2)IM, C(O)-(CH2)IM- or C(O)-(CH2)IM-NRM, where IM and RM are the same as above. In some embodiments, ASPGRM group set forth above is represented as follows:
Figure imgf000033_0001
The term "neoplasia" or “cancer” is used throughout the specification to refer to the pathological process that results in the formation and growth of a cancerous or malignant neoplasm, i.e., abnormal tissue that grows by cellular proliferation, often more rapidly than normal and continues to grow after the stimuli that initiated the new growth cease. Malignant neoplasms show partial or complete lack of structural organization and functional coordination with the normal tissue and most invade surrounding tissues, metastasize to several sites, and are likely to recur after attempted removal and to cause the death of the patient unless adequately treated. As used herein, the term neoplasia is used to describe all cancerous disease states and embraces or encompasses the pathological process associated with malignant hematogenous, ascitic and solid tumors. Neoplasms include, without limitation, morphological irregularities in cells in tissue of a subject or host, as well as pathologic proliferation of cells in tissue of a subject, as compared with normal proliferation in the same type of tissue. Additionally, neoplasms include benign tumors and malignant tumors (e.g., colon tumors) that are either invasive or noninvasive. Malignant neoplasms (cancer) are distinguished from benign neoplasms in that the former show a greater degree of anaplasia, or loss of differentiation and orientation of cells, and have the properties of invasion and metastasis. Examples of neoplasms or neoplasias from which the target cell of the present disclosure may be derived include, without limitation, carcinomas (e.g., squamous-cell carcinomas, adenocarcinomas, hepatocellular carcinomas, and renal cell carcinomas), particularly those of the bladder, bowel, breast, cervix, colon, esophagus, head, kidney, liver, lung, neck, ovary, pancreas, prostate, and stomach; leukemias; benign and malignant lymphomas, particularly Burkitt's lymphoma and Non-Hodgkin's lymphoma; benign and malignant melanomas; myeloproliferative diseases; sarcomas, particularly Ewing's sarcoma, hemangiosarcoma, Kaposi's sarcoma, liposarcoma, myosarcomas, peripheral neuroepithelioma, and synovial sarcoma; tumors of the central nervous system (e.g., gliomas, astrocytomas, oligodendrogliomas, ependymomas, gliobastomas, neuroblastomas, ganglioneuromas, gangliogliomas, medulloblastomas, pineal cell tumors, meningiomas, meningeal sarcomas, neurofibromas, and Schwannomas); germ-line tumors (e.g., bowel cancer, breast cancer, prostate cancer, cervical cancer, uterine cancer, lung cancer, ovarian cancer, testicular cancer, thyroid cancer, astrocytoma, esophageal cancer, pancreatic cancer, stomach cancer, liver cancer, colon cancer, and melanoma); mixed types of neoplasias, particularly carcinosarcoma and Hodgkin's disease; and tumors of mixed origin, such as Wilms' tumor and teratocarcinomas (Beers and Berkow (eds.), The Merck Manual of Diagnosis and Therapy, 17.sup.th ed. (Whitehouse Station, N.J.: Merck Research Laboratories, 1999) 973-74, 976, 986, 988, 991). All of these neoplasms may be treated using compounds according to the present disclosure.
Representative common cancers to be treated with compounds according to the present disclosure include, for example, prostate cancer, metastatic prostate cancer, stomach, colon, rectal, liver, pancreatic, lung, breast, cervix uteri, corpus uteri, ovary, testis, bladder, renal, brain/CNS, head and neck, throat, Hodgkin’s disease, non-Hodgkin’s lymphoma, multiple myeloma, leukemia, melanoma, non-melanoma skin cancer, acute lymphocytic leukemia, acute myelogenous leukemia, Ewing’s sarcoma, small cell lung cancer, choriocarcinoma, rhabdomyosarcoma, Wilms’ tumor, neuroblastoma, hairy cell leukemia, mouth/pharynx, oesophagus, larynx, kidney cancer and lymphoma, among others, which may be treated by one or more compounds according to the present disclosure. Because of the activity of the present compounds, the present disclosure has general applicability treating virtually any cancer in any tissue, thus the compounds, compositions and methods of the present disclosure are generally applicable to the treatment of cancer and in reducing the likelihood of development of cancer and/or the metastasis of an existing cancer.
In certain particular aspects of the present disclosure, the cancer which is treated is metastatic cancer, a recurrent cancer or a drug resistant cancer, especially including a drug resistant cancer. Separately, metastatic cancer may be found in virtually all tissues of a cancer patient in late stages of the disease, typically metastatic cancer is found in lymph system/nodes (lymphoma), in bones, in lungs, in bladder tissue, in kidney tissue, liver tissue and in virtually any tissue, including brain (brain cancer/tumor). Thus, the present disclosure is generally applicable and may be used to treat any cancer in any tissue, regardless of etiology. The term “tumor” is used to describe a malignant or benign growth or tumefacent.
The term “autoimmune disease” refers to a disease or illness that occurs when the body tissues are attacked by its own immune system. The immune system is a complex organization within the body that is designed normally to "seek and destroy" invaders of the body, including infectious agents. In diseases which are described as autoimmune diseases, MIF levels are often elevated. The present disclosure seeks to inhibit or lower elevated MIF levels in patients with autoimmune disease (as well as inflammatory diseases and conditions and cancer) and by decreasing MIF levels, ameliorate many of the symptoms and secondary effects of these disease states and conditions. Examples of autoimmune diseases which often exhibit high expressed levels of MIF including, for example, systemic lupus erythematosus, Sjogren syndrome, Hashimoto thyroiditis, rheumatoid arthritisjuvenile (type 1) diabetes, polymyositis, scleroderma, Addison's disease, vtiligo, pernicious anemia, glomerulonephritis, and pulmonary fibrosis, among numerous others.
A more complete list of autoimmune diseases which may be treated by compounds and pharmaceutical compositions according to the present disclosure includes Addison's Disease, Autoimmune polyendodrine syndrome (APS) types 1, 2 and 3, autoimmune pancreatitis (AIP), diabetes mellitus type 1, autoimmune thyroiditis, Ord's thyroiditis, Grave's disease, autoimmune oophoritis, endometriosis, autoimmune orchitis, Sjogren's syndrome, autoimmune enteropathy, coeliac disease, Crohns' disease, microscopic colitis, ulcerative colitis, autophospholipid syndrome (APIS), aplastic anemia, autoimmune hemolytica anemia, autoimmune lymphoproliferative syndrome, autoimmune neutropenia, autoimmune thrombocytopenic purpura, cold agglutinin disease, essential mixed cryoglulinemia, Evans sndrome, pernicious anemia, pure red cell aplasia, thrombocytopenia, adiposis dolorosa, adult-onset Still's disease, anklyosing spondylitis, CREST syndrome, drug-induced lupus, enthesitis-related arthritis, esosiniphilic fasciitis, Felty syndrome, AgG4-related disease, juvenile arthritis, Lyme disease (chronic), mixed connective tissue dease (MCTD), palindromic rheurmatism, Parry Romberg syndrome, Parsonage-Turner syndrome, psoriatic arthritis, reactive arthritis, relapsing polychondritis, retroperitoneal fibrosis, rheumatic fever, rheumatoid arthritis, sarcoidosis, Schnitzler syndrome, systemic lupus erythematosus, undifferentiated connective tissue disease (UCTD), dematomy ositis, fibromyalgia, myositis, inclusion body myositis, myasthenia gravis, neuromyotonia, paraneoplastic cerebellar degeneration, polymysositis, acute disseminated encephalomyelitis (ADEM), acute motor axonic neuropathy, anti-NMDA receptor encephalitis, Balo concentric sclerosis, Bickerstaffs encephalitis, chronic inflamatory demyelinating polyneuropathy, Guillain-Barre syndome, Hashimoto's encephalopathy, idiopathic inflammatory demyelinating diseases, Lambert- Eaton myasthenic syndrome, mutiple sclerosis, pattern II, Oshtoran Syndrome, Pendiatric Autoimmune Neuropsychiatric Disorder Associated with Streptococcus (PANDAS), progressive inflammatory neuropathy, restless leg syndrome, stiff person syndrome, Syndenham chorea, transverse myelitis, autoimmune retinopathy, autoimmune uveitis, Cogan syndrome, Graves ophthalmopathy, intermediate uveitis, ligneous conjunctivitis, Mooren's ulcer, neuromyelitis optica, opsoclonus myoclonus syndrome, optic neuritis, scleritis, Susac's syndrome, sympathetic ophthalmia, Tolosa-Hunt syndrome, autoimmune inner ear disease (AIED), Meniere's disease, Behcet's disease, Eosiniphilic granulomatosis with polyangiitis (EGPA), giant cell arteritis, granulomatosis with polyangiitis (GPA), IgA vasculitis (IgAV), Kawasaki's disease, leukocytoclastic vasculitis, lupus vasculitis, rheumatoid vasculitis, microscopic polyangiitis (MPA), polyarteritis nodosa (PAN), polymyalgia rheumatica, urticarial vasculitis, vasculitis, primary immune deficiency, chronic fatigue syndrome, complex regional pain syndrome, eosiniphilic esopagitis, gastritis, interstitial lung disease, POEMS syndrome, Raynaud's syndrome, primary immunodeficiency and pyoderma gangrenosum, among others.
The term “inflammatory disease” is used to describe a disease or illness with acute, but more often chronic inflammation as a principal manifestation of the disease or illness. Inflammatory diseases include diseases of neurodegeneration (including, for example, Alzheimer’s disease, Parkinson’s disease, Huntington’s disease; other ataxias), diseases of compromised immune response causing inflammmation (e.g., dysregulation of T cell maturation, B cell and T cell homeostasis, counters damaging inflammation), chronic inflammatory diseases including, for example, inflammatory bowel disease, including Crohn’s disease, rheumatoid arthritis, lupus, multiple sclerosis, chronic obstructive pulmony disease/COPD, pulmonary fibrosis, cystic fibrosis, Sjogren’s disease; hyperglycemic disorders, diabetes (I and II), affecting lipid metabolism islet function and/or structure, pancreatic P-cell death and related hyperglycemic disorders, including severe insulin resistance, hyperinsulinemia, insulin-resistant diabetes (e.g. Mendenhall's Syndrome, Werner Syndrome, leprechaunism, and lipoatrophic diabetes) and dyslipidemia (e.g. hyperlipidemia as expressed by obese subjects, elevated low-density lipoprotein (LDL), depressed high- density lipoprotein (HDL), elevated triglycerides and metabolic syndrome, liver disease, renal disease (apoptosis in plaques, glomerular disease), cardiovascular disease (especially including infarction, ischemia, stroke, pressure overload and complications during reperfusion), muscle degeneration and atrophy, low grade inflammation, gout, silicosis, atherosclerosis and associated conditions such as cardiac and neurological (both central and peripheral) manifestations including stroke, age-associated dementia and sporadic form of Alzheimer's disease, and psychiatric conditions including depression), stroke and spinal cord injury, arteriosclerosis, among others. In these diseases, elevated MIF is very often observed, making these disease states and/or conditions response to therapy using compounds and/or pharmaceutical compositions according to the present disclosure. It is noted that there is some overlap between certain autoimmune diseases and inflammatory diseases as described herein.
The term “linker”, refers to a chemical entity including a complex linker connecting a macrophage migration inhibitory factor binding moiety (MIFBM) to the asialoglycoprotein receptor binding moiety (ASGPRBM), optionally through at least one (such as one or two) connector moiety [CON] through covalent bonds in compounds according to the present disclosure. The linker between the two active portions of the molecule, that is the MIFBM and the ASGPRBM ranges from about 5 A to about 50A or more in length, about 6A to about 45 A in length, about 7 A to about 40 A in length, about 8 A to about 35 A in length, about 9 A to about 30A in length, about 10A to about 25 A in length, about 7 A to about 20 A in length, about 5 A to about 16A in length, about 5 A to about 15A in length, about 6 A to about 14A in length, about 10A to about 20 A in length, about 11 A to about 25 A in length, etc. Linkers which are based upon ethylene glycol units and are between 2 and 15 glycol units, 1 and 8 glycol units, 1, 2, 3, 4, 5, and 6 glycol units in length may be used, although the length of certain linkers may be far greater. By having a linker with a length as otherwise disclosed herein, the MIFBM group and the ASGPRBM group may be situated to advantageously take advantage of the biological activity of compounds according to the present disclosure which bind to asialoglycoprotein receptors on hepatocytes resulting in the selective and targeted degradation of MIF circulating proteins within the lysosomal degradation mechanism or other degradation mechanism of the hepatocytes. The selection of a linker component is based on its documented properties of biocompatibility, solubility in aqueous and organic media, and low immunogenicity/antigenicity. Although numerous linkers may be used as otherwise described herein, a linker based upon polyethyleneglycol (PEG) linkages, polypropylene glycol linkages, or poly ethyl eneglycol-co-polypropylene oligomers (up to about 100 units, about 1 to 100, about 1 to 75, about 1 to 60, about 1 to 50, about 1 to 35, about 1 to 25, about 1 to 20, about 1 to 15, 2 to 10, about 4 to 12, about 1 to 8, 1 to 3, 1 to 4, 2 to 6, 1 to 5, etc.) may be favored as a linker because of the chemical and biological characteristics of these molecules. Polyethylene (PEG) linkages of between 2 and 15 ethylene glycol units are sometimes used. When describing linkers according to the present disclosure, including polyethylene glycol linkers or other linkers, one or more additional groups (e.g., methylene groups, amide groups, keto groups, amine groups, etc.) may be covalently attached at either end of the linker group to attach to a ASGPRBM group, a [CON] group, another linker group or a CPBM group.
Alternative linkers may include, for example, polyamino acid linkers of up to 100 amino acids (of any type, such as D- or L- amino acids, such as naturally occurring L-amino acids) in length (about 1 to 75, about 1 to 60, about 1 to 50, about 1 to 45, about 1 to 35, about 1 to 25, about 1 to 20, about 1 to 15, 2 to 10, about 4 to 12, about 5 to 10, about 4 to 6, about 1 to 8, about 1 to 6 , about 1 to 5, about 1 to 4, about 1 to 3, etc. in length), optionally including one or more connecting groups (such as 1 or 2 connecting groups at one or both ends of the polyamino acid linker).
Exemplary linkers include those according to the chemical structures:
Figure imgf000038_0001
or a polypropylene glycol or polypropylene-co-polyethylene glycol linker having between 1 and 100 alkylene glycol units, such as about 1 to 75, about 1 to 60, about 1 to 50, about 1 to 45, about 1 to 35, about 1 to 25, about 1 to 20, about 1 to 15, 2 to 10, about 4 to 12, about 5 to 10, about 4 to 6, about 1 to 8, about 1 to 6, about 1 to 5, about 1 to 4, about 1 to 3; where Ra is H, C1-C3 alkyl or alkanol or forms a cyclic ring with R3 (proline) and R3 is a side chain derived from a D- or L amino acid (such as a naturally occurring L-amino acid), such as selected from the group consisting of alanine (methyl), arginine (propyleneguanidine), asparagine (methylenecarboxyamide), aspartic acid (ethanoic acid), cysteine (thiol, reduced or oxidized di-thiol), glutamine (ethylcarboxyamide), glutamic acid (propanoic acid), glycine (H), histidine (methyleneimidazole), isoleucine (1 -methylpropane), leucine (2 -methylpropane), lysine (butyleneamine), methionine (ethylmethylthioether), phenylalanine (benzyl), proline hydroxyproline (R3 forms a cyclic ring with Ra and the adjacent nitrogen group to form a pyrrolidine or hydroxypyrrolidine group), serine (methanol), threonine (ethanol, 1 -hydroxy ethane), tryptophan (m ethyleneindole), tyrosine (methylene phenol) or valine (isopropyl); m (within the context of this use) is an integer ranging from 1 to 100, such as from 1 to 75, 1 to 60, 1 to 55, 1 to 50, 1 to 45, 1 to 40, 2 to 35, 3 to 30, 1 to 15, 1 to 12, 1 to 10, 1 to 8, I to 6, 1, 2, 3, 4 or 5.
Other exemplary linkers include a polyethylene glycol linker containing from 1 to 1 to 100, such as 1 to 75, 1 to 60, 1 to 55, 1 to 50, 1 to 45, 1 to 40, 2 to 35, 3 to 30, 1 to 15, 1 to 10, 1 to 8, 1 to 6, 1, 2, 3, 4 or 5 ethylene glycol units, to which is bonded a lysine group or other amino acid moiety at one or both ends of the linker (which can consist of between 1 and 10 amino acids which can bind the MIFBM and/or the ASGPRBM group. Still other linkers comprise amino acid residues (D or L) which are bonded to MIFBM and/or ASGPRBM moieties as otherwise described herein. In another embodiment, as otherwise described herein, the amino acid has anywhere from 1-15 methylene groups separating the amino group from the acid (acyl) group in providing a linker to the MIFBM and/or the ASGPRBM group, wherein the linker contains from 1 to 100, 1 to 75, 1 to 60, 1 to 55, 1 to 50, 1 to 45, 1 to 40, 2 to 35, 3 to 30, 1 to 15, 1 to 10, 1 to 8, 1 to 6, 1, 2, 3, 4 or 5 amino acid groups linked together through peptide linkages to form the linker. This linker is represented by the chemical structure:
Figure imgf000039_0001
where Ram is H or a C1-C3 alkyl optionally substituted with one or two hydroxyl groups; na is an integer ranging from 1 to 15, such as 1-12, 1-10, 1-8, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11; m is an integer ranging from 1 to 100, such as 1 to 75, 1 to 60, 1 to 55, 1 to 50, 1 to 45, 1 to 40, 2 to 35, 3 to 30, 1 to 15, 1 to 12, 1 to 10, 1 to 8, 1 to 6, 1, 2, 3, 4 or 51 to 50, 1 to 45, 1 to 40, 2 to 35, 3 to 30, 1 to 15, 1 to 10, 1 to 8, 1 to 6, 1, 2, 3, 4 or 5. or another linker is according to the chemical formula:
Figure imgf000039_0002
where Z and Z’ are each independently a bond, -(CH2)i-O, -(CH2)i-S, -(CH2)i-N-R,
Figure imgf000039_0003
wherein said -(CH2)i group, if present in Z or Z’, is bonded to a connector (CON), MIFBM or ASGPRBM; each R is H, or a C1-C3 alkyl or alkanol group; each R2 is independently H or a C1-C3 alkyl group; each Y is independently a bond, O, S or N-R; each i is independently an integer ranging from 0 to 100, such as 0 to 75, 1 to 60, 1 to 55, 1 to 50, 1 to 45, 1 to 40, 2 to 35, 3 to 30, 1 to 15, 1 to 10, 1 to 8, 1 to 6, 0, 1, 2, 3, 4 or 5;
D is
Figure imgf000040_0001
a bond, with the proviso that Z, Z’ and D are not each simultaneously bonds; j is an integer ranging from 1 to 100, such as 1 to 75, 1 to 60, 1 to 55, 1 to 50, 1 to 45,
1 to 40, 2 to 35, 3 to 30, 1 to 15, 1 to 10, 1 to 8, 1 to 6, 1, 2, 3, 4 or 5; m’ is an integer ranging from 1 to 100, such as 1 to 75, 1 to 60, 1 to 55, 1 to 50, 1 to
45, 1 to 40, 2 to 35, 3 to 30, 1 to 15, 1 to 10, 1 to 8, 1 to 6, 1, 2, 3, 4 or 5; n is an integer ranging from 1 to 100, such as 1 to 75, 1 to 60, 1 to 55, 1 to 50, 1 to 45, 1 to 40, 2 to 35, 3 to 30, 1 to 15, 1 to 10, 1 to 8, 1 to 6, 1, 2, 3, 4 or 5;
X1 is O, S or N-R; and
R is H, or a C1-C3 alkyl or alkanol group, or a pharmaceutical salt thereof.
Other linkers which are included herein include linkers according to the chemical structure:
Figure imgf000040_0002
where each n and n’ is independently an integer ranging from 1 to 25, such as 1 to 15, 1 to 12, 2 to 11, 2 to 10, 2 to 8, 2 to 6, 2 to 5, 2 to 4 and 2 to 3 or 1, 2, 3, 4, 5, 6, 7, or 8; and each n” is independently an integer ranging from 0 to 8, such as 1 to 7, or 1, 2, 3, 4, 5 or 6.
Linkers also can comprise two or more linker segments (based upon the linkers described above) which are attached directly to each other or through [CON] groups forming a complex linker. Certain linkers which include a [CON] group (especially a diamide [CON] group as otherwise described herein) connecting a first and second (PEG) linker group include the following structures:
Figure imgf000041_0001
where each n and n’ is independently an integer ranging from 1 to 25, such as 1 to 15, 1 to 12, 2 to 11, 2 to 10, 2 to 8, 2 to 6, 2 to 5, 2 to 4 and 2 to 3 or 1, 2, 3, 4, 5, 6, 7, or 8; and each n” is independently an integer ranging from 0 to 8, such as 1 to 7, or 1, 2, 3, 4, 5 or 6. Noted is that each of these linkers may also contain alkylene groups containing from 1 to 4 methylene groups at the distal ends of each linker group in order to facilitate connection of the linker group.
Other linkers which include a connector group [CON] include groups which are represented by the chemical formula PEG-[CON]-PEG wherein each PEG is independently a polyethylene glycol group containing from 1-12 ethylene glycol residues and [CON] is a connector group as otherwise set forth herein, such as a triazole group
Figure imgf000041_0002
The term “connector”, symbolized in the generic formulas by “CON” or [CON], is used to describe a chemical moiety which is optionally included in bifunctional compounds according to the present disclosure which forms from the reaction product of an activated linker with a MIFBM moiety (which also is sometimes activated for covalently bonding the linker with the moiety) or a ASGPRBM group with an activated linker. The connector group is often the resulting moiety which forms from the facile condensation of two or more separate chemical fragments which contain reactive groups which can provide connector groups as otherwise described to produce bifunctional or multifunctional compounds according to the present disclosure. It is noted that a connector may be distinguishable from a linker in that the connector is the result of a specific chemistry which is used to provide bifunctional compounds according to the present disclosure wherein the reaction product of these groups results in an identifiable connector group or part of a connector group which is distinguishable from the linker group, although in certain instances, the connector group is incorporated into and integral with the linker group as otherwise described herein. It is noted also that a connector group may be linked to a number of linkers to provide multifunctionality (i.e., more than one CPBM moiety and/or more than one ASGPRBM/FCRNBM moiety) within the same molecule. It is noted that there may be some overlap between the description of the connector group and the linker group such that the connector group is actually incorporated or forms part of the linker, especially with respect to more common connector groups such as amide groups, oxygen (ether), sulfur (thioether) or amine linkages, urea or carbonate -OC(O)O- groups or as otherwise described herein. It is further noted that a connector (or linker) may be connected to MIFBM, ASGPRBM or a linker at positions which are represented as being linked to another group using the symbol
Figure imgf000042_0001
Where two or more such groups are present in a linker or connector, any of an ASGPRBM, a linker or a MIFBM may be bonded to such a group. Where that symbol is not used, the linker may be at one or more positions of a moiety.
Common connector groups which are used in the present disclosure include the following chemical groups:
Figure imgf000042_0002
where RCON1 and RC0N2 are each independently H, methyl or a bond (for attachment to another moiety); or a diamide group according to the structure:
Figure imgf000043_0001
where X2 is CH2, O, S, NR4, C(O), S(O), S(O)2, -S(O)2O, -OS(O)2, or OS(O)2O;
X3 is O, S, NR4;
R4 is H, a C1-C3 alkyl or alkanol group, or a -C(O)(Ci-C3) group;
Ri is H or a C1-C3 alkyl group; and n” is independently an integer ranging from 0 to 8, such as 1 to 7, or 1, 2, 3, 4, 5 or 6; or the connector group [CON] is a group according to the chemical structure:
Figure imgf000043_0002
where R1C0N, R2CON and R3C0N are each independently H, -(CH2)MCI, -
(CH2)MClaC(O)xA(NR4)xA-(CH2)MCla ,-(CH2)MCla(NR4)xAC(O)xA-(CH2)MCla Or -(CH2)MClaO-
(CH2)MCI-C(O)NR4-, with the proviso that R1C0N, R2C0N and R3C0N are not simultaneously H; each MCI is independently an integer ranging from 1 to 4, such as 1 or 2; each MCI a is independently an integer ranging from 0 to 4, such as 0, 1 or 2; and R4 is H, a C1-C3 alkyl or alkanol group, or a -C(O)(Ci-C3) group.
The triazole group, indicated above, is an exemplary connector group. An additional exemplary connector group is
Figure imgf000043_0003
which is linked to at least one MIFBM and/or at least one ASPRGBM (such as 3 ASPRGBM moieties). This connector group may be used to form GN3.
It is noted that each connector may be extended with one or more methylene groups to facilitate connection to a linker group, another CON group, a MIFBM group or a ASGPRBM. It is noted that in certain instances, within context the diamide group may also function independently as a linker group. Additional Galactose- and Talose-based ASGPR Binding Moieties
In certain embodiments, the present disclosure is directed to compounds which are useful for removing circulating proteins which are associated with a disease state or condition in a patient or subject according to the general chemical structure of Formula II:
Figure imgf000044_0001
Formula II
The term "Extracellular Protein Targeting Ligand" as used herein is interchangeably used with the term CPBM (cellular protein binding moiety). The term "ASGPR Ligand" as used herein is interchangeably used with an asialoglycoprotein receptor (ASGPR) binding moiety as defined herein.
In the compound of Formula II, each [CON] is an optional connector chemical moiety which, when present, connects directly to [CPBM] or to [CRBM] or connects the [LINKER- 2] to [CPBM] or to [CRBM],
In the compound of Formula II:
[LINKER-2] is a chemical moiety having a valency from 1 to 15 which covalently attaches to one or more [CRBM] and/or [CPBM] group, optionally through a [CON], including a [MULTICON] group, wherein said [LINKER-2] optionally itself contains one or more [CON] or [MULTICON] group(s); k’ is an integer from 1 to 15; j’ is an integer from 1 to 15; h and h’ are each independently an integer from 0 to 15;
IL is an integer from 0 to 15; with the proviso that at least one of h, h’ and 11 is at least 1, or a pharmaceutically acceptable salt, stereoisomer, solvate or polymorph thereof.
A [MULTICON] group can connect one or more of a [CRBM] or [CPBM] to one or more of a [LINKER-2], In various embodiments, [LINKER-2] has a valency of 1 to 10. In various embodiments, [LINKER-2] has a valency of 1 to 5. In various embodiments, [LINKER-2] has a valency of 1, 2 or 3. In various embodiments, in the compound of Formula II, the [LINKER-2] includes one or more of Linker^ Linker®, Linker13, Linker0, and/or combinations thereof as defined herein.
In the compound of Formula II, xx is independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, and 25.
In the compound of Formula II, yy is independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, and 25.
In the compound of Formula II, zz is independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, and 25.
In the compound of Formula II, X1 is 1 to 5 contiguous atoms independently selected from O, S, N(Rb), and C(R4)(R4), wherein if X1 is 1 atom then X1 is O, S, N(R6), or C(R4)(R4), if X1 is 2 atoms then no more than 1 atom of X1 is O, S, or N(R6), if X1 is 3, 4, or 5 atoms then no more than 2 atoms of X1 are O, S, or N(R6);
R3 at each occurrence is independently selected from hydrogen, alkyl, heteroalkyl, haloalkyl (including -CF3, -CHF2, -CH2F, -CH2CF3, -CH2CH2F, and -CF2CF3), arylalkyl, heteroarylalkyl, alkenyl, alkynyl, and, heteroaryl, heterocycle, -OR8, and -NR8R9;
R4 is independently selected at each occurrence from hydrogen, heteroalkyl, alkyl, haloalkyl, arylalkyl, heteroarylalkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocycle, -OR6, - NR6R7,
R6 and R7 are independently selected at each occurrence from hydrogen, heteroalkyl, alkyl, arylalkyl, heteroaryl alkyl, alkenyl, alkynyl, and, haloalkyl, heteroaryl, heterocycle, - alkyl-OR8, -alkyl-NR8R9, C(O)R3, S(O)R3, C(S)R3, and S(O)2R3;
R8 and R9 are independently selected at each occurrence from hydrogen, heteroalkyl, alkyl, arylalkyl, heteroarylalkyl, alkenyl, alkynyl, aryl, heteroaryl, and heterocycle.
A. Galactose-Based ASGPR-Binding Cellular Receptor Binding Moieties of Formula II
In certain embodiments, the compound of Formula II is selected from:
Figure imgf000045_0001
Figure imgf000046_0001
Figure imgf000047_0001
Figure imgf000048_0001
Figure imgf000049_0001
Figure imgf000050_0001
Figure imgf000051_0001
Figure imgf000052_0001
Figure imgf000053_0001
Figure imgf000054_0001
Figure imgf000055_0001
Figure imgf000056_0001
Figure imgf000057_0001
Figure imgf000058_0001
Figure imgf000059_0001
Figure imgf000060_0001
Figure imgf000061_0001
Figure imgf000062_0001
In certain embodiments, the compound of Formula II has one of the following structures:
Figure imgf000062_0002
Figure imgf000063_0001
In various embodiments, the ASGPR ligand is linked at either the C1 or C5 (R1 or R5) position to form a degrading compound. In various embodiments, the ASGPR ligand is linked at C6 position to form a degrading compound. For example, when the ASGPR ligand is
Figure imgf000063_0002
then non- limiting examples of ASGPR binding compounds of Formula II include:
Figure imgf000063_0003
or the bi- or tri- substituted versions thereof or pharmaceutically acceptable salts thereof, where the bi- or tri- substitution refers to the number additional galactose derivatives attached to a linker moiety.
In any of the embodiments herein where an ASGPR ligand is drawn for use in a degrader the ASGPR ligand is typically linked through to the Extracellular Protein Targeting Ligand in the C5 position (e.g., which can refer to the adjacent C6 carbon hydroxyl or other functional moiety that can be used for linking purposes). When the linker and Extracellular Protein Targeting Ligand is connected through the C1 position, then that carbon is appropriately functionalized for linking, for example with a hydroxyl, amino, allyl, alkyne or hydroxyl-allyl group.
In various embodiments, the ASGPR ligand is not linked in the C3 or C4 position, because these positions chelate with the calcium for ASGPR binding in the liver. In certain embodiments, an ASGPR ligand useful for incorporation into a compound of Formula II is selected from:
Figure imgf000064_0001
Figure imgf000065_0001
Figure imgf000066_0001
Figure imgf000067_0001
Figure imgf000068_0001
Figure imgf000069_0001
Figure imgf000070_0001
Figure imgf000071_0001
Figure imgf000072_0001
Figure imgf000073_0001
Figure imgf000074_0001
Figure imgf000075_0001
In certain embodiments, the compound of Formula II is selected from:
Figure imgf000075_0002
Figure imgf000076_0001
Figure imgf000077_0001
Figure imgf000078_0001
Figure imgf000079_0001
B. Talose-Based ASGPR-Binding Cellular Receptor Binding Moieties of Formula II
5 In certain embodiments, the compound of Formula II is selected from:
Figure imgf000079_0002
Figure imgf000080_0001
Figure imgf000081_0001
Figure imgf000082_0001
Figure imgf000083_0001
Figure imgf000084_0001
Figure imgf000085_0001
Figure imgf000086_0001
Figure imgf000087_0001
Figure imgf000088_0001
Figure imgf000089_0001
Figure imgf000090_0001
Figure imgf000091_0001
Figure imgf000092_0001
Figure imgf000093_0001
Figure imgf000094_0001
Figure imgf000095_0001
Figure imgf000096_0001
Figure imgf000097_0001
S ' | _ . Q
Figure imgf000098_0001
Figure imgf000099_0001
Figure imgf000100_0001
Figure imgf000101_0001
Figure imgf000102_0001
Figure imgf000103_0001
Figure imgf000104_0001
Figure imgf000105_0001
Figure imgf000106_0001
Figure imgf000107_0001
Figure imgf000108_0001
Figure imgf000109_0001
Figure imgf000110_0001
<0^ fSiss^n SWfW
SW&M WWW3
Figure imgf000111_0001
Figure imgf000112_0001
Figure imgf000113_0001
ʼnll
Figure imgf000114_0001
Figure imgf000115_0001
Figure imgf000116_0001
In certain embodiments, the compound of Formula II is an Extracellular Protein degrading compound in which the ASGPR ligand is a ligand as described herein
Figure imgf000116_0002
In certain embodiments, in the compound of Formula II, the ASGPR ligand is linked at either the Cl or C5 (R1 or R5) position to form a degrading compound. In certain embodiments, in the compound of Formula II, the ASGPR ligand is linked at C6. In various embodiments, when the ASGPR ligand is
Figure imgf000116_0003
then non- limiting examples of ASGPR binding compounds of Formula II include:
Figure imgf000117_0001
or the bi- or tri- substituted versions thereof or pharmaceutically acceptable salts thereof, where the bi- or tri- substitution refers to the number additional galactose derivatives attached to a linker moiety. In certain embodiments the compound of Formula II is selected from:
Figure imgf000118_0001
wherein in certain embodiments R2 is selected from -NR6COR3, -NR6-(5-membered heteroaryl), and-NR6-(6-membered heteroaryl), each of which R2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
In certain embodiments, the compound of Formula II is selected from:
Figure imgf000119_0001
Figure imgf000120_0001
wherein in certain embodiments R2 is selected from -NR6COR3, -NR6-(5-membered heteroaryl), and-NR6-(6-membered heteroaryl), each of which R2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
In certain embodiments, the compound of Formula II is selected from:
Figure imgf000121_0001
Figure imgf000122_0001
wherein in certain embodiments R2 is selected from -NR6COR3, -NR6-(5-membered heteroaryl), and-NR6-(6-membered heteroaryl), each of which R2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
In certain embodiments, the compound of Formula II is selected from:
Figure imgf000123_0001
Figure imgf000124_0001
wherein in certain embodiments R2 is selected from -NR6COR3, -NR6-(5-membered heteroaryl), and-NR6-(6-membered heteroaryl), each of which R2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
In certain embodiments, the compound of Formula II is selected from:
Figure imgf000125_0001
Figure imgf000126_0001
wherein in certain embodiments R2 is selected from -NR6COR3, -NR6-(5-membered heteroaryl), and-NR6-(6-membered heteroaryl), each of which R2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
In certain embodiments, the compound of Formula II is selected from:
Figure imgf000127_0001
Figure imgf000128_0001
wherein in certain embodiments R2 is selected from -NR6COR3, -NR6-(5-membered heteroaryl), and-NR6-(6-membered heteroaryl), each of which R2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
In certain embodiments, the compound of Formula II is selected from:
Figure imgf000129_0001
Figure imgf000130_0001
wherein in certain embodiments R2 is selected from -NR6COR3, -NR6-(5-membered heteroaryl), and-NR6-(6-membered heteroaryl), each of which R2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
In certain embodiments, the compound of Formula II is selected from:
Figure imgf000131_0001
Figure imgf000132_0001
wherein in certain embodiments R2 is selected from -NR6COR10, -NR6-(5-membered heteroaryl), and-NR6-(6-membered heteroaryl), each of which R2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
In certain embodiments, the compound of Formula II is selected from:
Figure imgf000133_0001
Figure imgf000134_0001
wherein in certain embodiments R2 is selected from -NRbCOR10, -NR6-(5-membered heteroaryl), and-NR6-(6-membered heteroaryl), each of which R2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
In certain embodiments, the compound of Formula II is selected from:
Figure imgf000135_0001
Figure imgf000136_0001
Figure imgf000137_0001
wherein in certain embodiments R2 is selected from -NR6COR10, -NR6-(5-membered heteroaryl), and-NR6-(6-membered heteroaryl), each of which R2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
In certain embodiments, the compound of Formula II is selected from:
Figure imgf000138_0001
Figure imgf000139_0001
wherein in certain embodiments R2 is selected from -NR6COR10, -NR6-(5-membered heteroaryl), and-NR6-(6-membered heteroaryl), each of which R2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
In certain embodiments, the compound of Formula II is selected from:
Figure imgf000140_0001
Figure imgf000141_0001
wherein in certain embodiments R2 is selected from -NR6COR10, -NR6-(5-membered heteroaryl), and-NR6-(6-membered heteroaryl), each of which R2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
In certain embodiments, the compound of Formula II is selected from:
Figure imgf000142_0001
Figure imgf000143_0001
wherein in certain embodiments R2 is selected from -NR6COR10, -NR6-(5-membered heteroaryl), and-NR6-(6-membered heteroaryl), each of which R2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
In certain embodiments, the compound of Formula II is selected from:
Figure imgf000144_0001
Figure imgf000145_0001
wherein in certain embodiments R2 is selected from -NR6COR10, -NR6-(5-membered heteroaryl), and-NR6-(6-membered heteroaryl), each of which R2 groups is optionally substituted with 1, 2, 3, or 4 independent, substituents as described herein, for example 1, 2, 3, or 4 substituents independently selected from F, Cl, Br, haloalkyl, or alkyl.
In certain embodiments, the compound of Formula II is selected from:
Figure imgf000146_0001
Figure imgf000147_0001
Figure imgf000148_0001
Figure imgf000149_0001
Figure imgf000150_0001
Figure imgf000151_0001
Figure imgf000152_0001
Figure imgf000153_0001
Figure imgf000154_0001
Figure imgf000155_0001
Figure imgf000156_0001
In certain embodiments, an ASGPR ligand useful for incorporation into a compound of Formula II is selected from:
Figure imgf000156_0002
Figure imgf000157_0001
Figure imgf000158_0001
Figure imgf000159_0001
Figure imgf000160_0001
Figure imgf000161_0001
Figure imgf000162_0001
Figure imgf000163_0001
Figure imgf000164_0001
Figure imgf000165_0001
Figure imgf000166_0001
Figure imgf000167_0001
Figure imgf000168_0001
5 C. The ASGPR Ligand/Binding Moiety in Compounds of Formula II
In certain embodiments, in the compound of Formula II, R1 is hydrogen.
Figure imgf000169_0001
In certain embodiments, in the compound of Formula II, R1 is
Figure imgf000169_0002
In certain embodiments, in the compound of Formula II, R1 is
Figure imgf000169_0003
In certain embodiments, in the compound of Formula II, R1 is
Figure imgf000169_0004
In certain embodiments, in the compound of Formula II, R1 is
Figure imgf000169_0005
In certain embodiments, in the compound of Formula II, R1 is
In certain embodiments, in the compound of Formula II, R1 is
Figure imgf000169_0006
In certain embodiments, in the compound of Formula II, R1 is Co-Cealkyl-cyano optionally substituted with 1, 2, 3, or 4 substituents.
In certain embodiments, in the compound of Formula II, R1 is alkyl optionally substituted with 1, 2, 3, or 4 substituents.
In certain embodiments, in the compound of Formula II, R1 is alkenyl optionally substituted with 1, 2, 3, or 4 substituents. In certain embodiments, in the compound of Formula II, R1 is alkynyl optionally substituted with 1, 2, 3, or 4 substituents. In certain embodiments, in the compound of Formula II, R1 is haloalkyl optionally substituted with 1, 2, 3, or 4 substituents. In certain embodiments, in the compound of Formula II, R1 is F.
In certain embodiments, in the compound of Formula II, R1 is Cl.
In certain embodiments, in the compound of Formula II, R1 is Br.
In certain embodiments, in the compound of Formula II, R1 is aryl optionally substituted with 1, 2, 3, or 4 substituents.
In certain embodiments, in the compound of Formula II, R1 is arylalkyl optionally substituted with 1, 2, 3, or 4 substituents.
In certain embodiments, in the compound of Formula II, R1 is heteroaryl optionally substituted with 1, 2, 3, or 4 substituents.
In certain embodiments, in the compound of Formula II, R1 is heteroaryl alkyl optionally substituted with 1, 2, 3, or 4 substituents.
In certain embodiments, in the compound of Formula II, R1 is heterocycle optionally substituted with 1, 2, 3, or 4 substituents.
In certain embodiments, in the compound of Formula II, R1 is heterocycloalkyl optionally substituted with 1, 2, 3, or 4 substituents. In certain embodiments, in the compound of Formula II, R1 is haloalkoxy optionally substituted with 1, 2, 3, or 4 substituents. In certain embodiments, in the compound of Formula II, R1 is -O-alkenyl, -O-alkynyl, C0-C6alkyl-OR6, C0-C6alkyl-SR6, C0-C6alkyl-NR6R7, C0-C6alkyl-C(O)R3, C0-C6alkyl-S(O)R3, C0-C6alkyl-C(S)R3, C0-C6alkyl-S(O)2R3, C0-C6alkyl-N(R8)-C(O)R3, C0-C6alkyl-N(R8)- S(O)R3, C0-C6alkyl-N(R8)-C(S)R3, C0-C6alkyl-N(R8)-S(O)2R3 C0-C6alkyl-O-C(O)R3, C0- C6alkyl-O-S(O)R3, C0-C6alkyl-O-C(S)R3, -N=S(O)(R3)2, C0-C6alkylN3, or C0-C6alkyl-O- S(O)2R3, each of which is optionally substituted with 1, 2, 3, or 4 substituents. In certain embodiments, in the compound of Formula II, R2 is aryl optionally substituted with 1, 2, 3, or 4 substituents. In certain embodiments, in the compound of Formula II, R2 is heterocycle optionally substituted with 1, 2, 3, or 4 substituents. In certain embodiments, in the compound of Formula II, R2 is heteroaryl containing 1 or 2 heteroatoms independently selected from N, O, and S optionally substituted with 1, 2, 3, or 4 substituents. In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000170_0001
In certain embodiments, in the compound of Formula II, R2 is heterocycle optionally substituted with 1, 2, 3, or 4 substituents. In certain embodiments, in the compound of Formula II, R2 is -NR8-S(O)-R3 optionally substituted with 1, 2, 3, or 4 substituents. In certain embodiments, in the compound of Formula II, R2 is -NR8-C(S)-R3 optionally substituted with 1, 2, 3, or 4 substituents. In certain embodiments, in the compound of Formula II, R2 is -NR8-S(O)(NR6)-R3 optionally substituted with 1, 2, 3, or 4 substituents. In certain embodiments, in the compound of Formula II, R2 is -N=S(O)(R3)2 optionally substituted with 1, 2, 3, or 4 substituents. In certain embodiments, in the compound of Formula II, R2 is -NR8C(O)NR9S(O)2R3 optionally substituted with 1, 2, 3, or 4 substituents.
In certain embodiments, in the compound of Formula II, R2 is -NR8-S(O)2-R10 optionally substituted with 1, 2, 3, or 4 substituents.
In certain embodiments, in the compound of Formula II, R2 is -NR8-C(NR6)-R3 optionally substituted with 1, 2, 3, or 4 substituents.
In certain embodiments, in the compound of Formula II, R2 is hydrogen.
In certain embodiments, in the compound of Formula II, R2 is R10,
In certain embodiments, in the compound of Formula II, R2 is alkyl-C(O)-R3.
In certain embodiments, in the compound of Formula II, R2 is -C(O)-R3.
In certain embodiments, in the compound of Formula II, R2 is alkyl.
In certain embodiments, in the compound of Formula II, R2 is haloalkyl.
In certain embodiments, in the compound of Formula II, R2 is -OC(O)R3.
In certain embodiments, in the compound of Formula II, R2 is -NR8-C(O)R10.
In certain embodiments, in the compound of Formula II, R2 is alkenyl optionally substituted with 1, 2, 3, or 4 substituents.
In certain embodiments, in the compound of Formula II, R2 is allyl optionally substituted with 1, 2, 3, or 4 substituents.
In certain embodiments, in the compound of Formula II, R2 is alkynyl optionally substituted with 1, 2, 3, or 4 substituents.
In certain embodiments, in the compound of Formula II, R2 is -NR6-alkenyl optionally substituted with 1, 2, 3, or 4 substituents.
In certain embodiments, in the compound of Formula II, R2 is -O-alkenyl optionally substituted with 1, 2, 3, or 4 substituents.
In certain embodiments, in the compound of Formula II, R2 is -NR6-alkynyl optionally substituted with 1, 2, 3, or 4 substituents.
In certain embodiments, in the compound of Formula II, R2 is -NR6 -heteroaryl optionally substituted with 1, 2, 3, or 4 substituents.
In certain embodiments, in the compound of Formula II, R2 is -NR6-aryl optionally substituted with 1, 2, 3, or 4 substituents.
In certain embodiments, in the compound of Formula II, R2 is -O-heteroaryl optionally substituted with 1, 2, 3, or 4 substituents.
In certain embodiments, in the compound of Formula II, R2 is -O-aryl optionally substituted with 1, 2, 3, or 4 substituents.
In certain embodiments, in the compound of Formula II, R2 is -O-alkynyl optionally substituted with 1, 2, 3, or 4 substituents.
In certain embodiments, in the compound of Formula II, R2 is selected from and
Figure imgf000172_0001
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000172_0002
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000172_0003
R is an optional substituent as defined herein. In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000173_0001
In certain embodiments, in the compound of Formula II, R2A is selected from
Figure imgf000173_0002
an optional substituent as defined herein.
In certain embodiments, in the compound of Formula II, R2A is selected from
Figure imgf000173_0003
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000174_0001
,
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000175_0001
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000175_0002
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000175_0003
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000176_0001
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000176_0002
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000176_0003
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000176_0004
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000176_0005
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000177_0001
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000177_0002
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000177_0003
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000177_0004
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000177_0005
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000178_0001
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000178_0002
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000178_0003
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000179_0001
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000179_0002
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000179_0003
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000180_0001
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000180_0002
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000180_0003
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000180_0004
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000180_0005
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000180_0006
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000180_0007
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000181_0001
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000181_0002
In certain embodiments, in the compound of Formula II, R2 or R2A is selected from
Figure imgf000181_0003
Figure imgf000182_0001
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000182_0002
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000182_0003
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000182_0004
In certain embodiments, in the compound of Formula II, R2 is selected from
Figure imgf000182_0005
In certain embodiments, in the compound of Formula II, R2 is selected from In certain embodiments, in the compound of Formula II, R2 is a spirocyclic
Figure imgf000183_0001
heterocycle, for example, and without limitation,
In certain embodiments, in the compound of Formula II, R2 is a silicon containing
Figure imgf000183_0002
heterocycle, for example, and without limitation, In certain embodiments, in the compound of Formula II, R2 is substituted with SFs,
Figure imgf000183_0003
for example, and without limitation,
In certain embodiments, in the compound of Formula II, R2 is substituted with a sulfoxime, for example, and without limitation,
Figure imgf000183_0004
In certain embodiments, in the compound of Formula II, R10 is selected from bicyclic heterocycle.
In certain embodiments, in the compound of Formula II, R10 is selected from spirocyclic heterocycle.
In certain embodiments, in the compound of Formula II, R10 is selected from -NR6- heterocycle.
In certain embodiments, in the compound of Formula II, R10 is selected from
Figure imgf000183_0005
In certain embodiments, in the compound of Formula II, R10 is selected from
Figure imgf000184_0001
In certain embodiments, in the compound of Formula II, R is selected from
Figure imgf000184_0002
In certain embodiments, in the compound of Formula II, R10 is selected from
Figure imgf000184_0003
In certain embodiments, in the compound of Formula II, Cycle is selected from
Figure imgf000184_0004
Figure imgf000185_0001
In certain embodiments, in the compound of Formula II, R30 is selected from:
Figure imgf000185_0002
In certain embodiments, in the compound of Formula II, R200 is
Figure imgf000185_0003
In certain embodiments, in the compound of Formula II, R is
In certain embodiments, in the compound of Formula II, R200 is
Figure imgf000185_0004
Figure imgf000186_0001
In certain embodiments, in the compound of Formula II, R200 is
In certain embodiments, in the compound of Formula II, R
Figure imgf000186_0002
is
In certain embodiments, in the compound of Formula II, R200 is
Figure imgf000186_0003
Figure imgf000186_0004
In certain embodiments, in the compound of Formula II, R200 is
In certain embodiments, in the compound of Formula II, R200 is
Figure imgf000186_0005
In certain embodiments, in the compound of Formula II, R200 is
Figure imgf000186_0006
Figure imgf000186_0007
In certain embodiments, in the compound of Formula II, R200 is :
In certain embodiments, in the compound of Formula II, R200 is
Figure imgf000186_0008
Figure imgf000186_0009
In certain embodiments, in the compound of Formula II, R200 is
Linkers
In non-limiting embodiments, in the compound of Formula II, LinkerA and Linker® are independently selected from:
Figure imgf000186_0010
wherein:
R11, R12, R13, R14, R15, R16, R17, R18, R19, and R20 are independently at each occurrence selected from the group consisting of a bond, alkyl, -C(O)-, -C(O)O-, -OC(O)-, -SO2-, -S(O)-, -C(S)-, -C(O)NR6-, -NR6C(O)-, -O-, -S-, -NR6-, -C(R21R21)-, -P(O)(R3)O-, -P(O)(R3)-, a divalent residue of a natural or unnatural amino acid, alkenyl, alkynyl, haloalkyl, alkoxy, and, heterocycle, heteroaryl, -CH2CH2-[O-(CH2)2]n-O-, CH2CH2-[O-(CH2)2]n-NR6-, -CH2CH2-[O- (CH2)2]n-, -[-(CH2)2-O-]n-, -[O-(CH2)2]n-,-[O-CH(CH3)C(O)]n-, -[C(O)-CH(CH3)-O]n-,
-[O-CH2C(O)]n-, -[C(O)-CH2-O]n -, a divalent residue of a fatty acid, a divalent residue of an unsaturated or saturated mono- or di-carboxylic acid; each of which is optionally substituted with 1, 2, 3, or 4 substituents independently selected from R21; n is independently selected at each instance from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;
R21 is independently at each occurrence selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, F, Cl, Br, I, hydroxyl, alkoxy, azide, amino, cyano, - NR6R7, -NR8SO2R3, -NR8S(O)R3, haloalkyl, heteroalkyl, and, heteroaryl, and heterocycle; and the remaining variables are as defined herein.
In certain embodiments, in the compound of Formula II, LinkerA is bond and Linker8 is
Figure imgf000187_0001
In certain embodiments, in the compound of Formula II, Linker8 is bond and Linked is
Figure imgf000187_0002
In certain embodiments, in the compound of Formula II, a divalent residue of an amino acid is selected from
Figure imgf000187_0003
Figure imgf000188_0001
wherein the amino acid can be oriented in either direction and wherein the amino acid can be in the L- or D-form or a mixture thereof. In certain embodiments, in the compound of Formula II, a divalent residue of a dicarboxylic acid is generated from a nucleophilic addition reaction:
Figure imgf000188_0002
Non-limiting embodiments of a divalent residue of a dicarboxylic acid generated from a nucleophilic addition reaction include:
Figure imgf000189_0001
In certain embodiments, in the compound of Formula II, a divalent residue of a dicarboxylic acid is generated from a condensation reaction:
Figure imgf000189_0002
Non-limiting embodiments of a divalent residue of a dicarboxylic acid generated from a condensation include:
Figure imgf000189_0003
Non-limiting embodiments of a divalent residue of a saturated dicarboxylic acid include:
Figure imgf000190_0001
Non-limiting embodiments of a divalent residue of a saturated dicarboxylic acid include:
Figure imgf000190_0002
Non-limiting embodiments of a divalent residue of a saturated monocarboxylic acid is selected from butyric acid (-OC(O)(CH2)2CH2-), caproic acid (-OC(O)(CH2)4CH2-), caprylic acid (-OC(O)(CH2)SCH2-), capric acid (-OC(O)(CH2)sCH2-), lauric acid (- OC(0)(CH2)IOCH2-), myristic acid (-OC(O)(CH2)i2CH2-), pentadecanoic acid (- OC(O)(CH2)i3CH2-), palmitic acid (-OC(O)(CH2)i4CH2-), stearic acid (-OC(O)(CH2)ieCH2-), behenic acid (-OC(0)(CH2)2oCH2-), and lignoceric acid (-OC(O)(CH2)22CH2-);
Non-limiting embodiments of a divalent residue of a fatty acid include residues selected from linoleic acid, palmitoleic acid, vaccenic acid, paullinic acid, oleic acid, elaidic acid, gondoic acid, gadoleic acid, nervonic acid, myristoleic acid, and erucic acid:
Figure imgf000191_0001
Non-limiting embodiments of a divalent residue of a fatty acid is selected from linoleic acid (-C(O)(CH2)?(CH)2CH2(CH)2(CH2)4CH2-), docosahexaenoic acid (-C(O)(CH2)2(CHCHCH2)eCH2-), eicosapentaenoic acid (- C(O)(CH2)3(CHCHCH2)5CH2-), alpha-linolenic acid (-C(O)(CH2)7(CHCHCH2)3CH2-) stearidonic acid
(-C(O)(CH2)4(CHCHCH2)4CH2-), y-linolenic acid (- C(O)(CH2)4(CHCHCH2)3(CH2)3CH2-), arachidonic acid (- C(O)(CH2)3,(CHCHCH2)4(CH2)4CH2-), docosatetraenoic acid (-C(O)(CH2)5(CHCHCH2)4(CH2)4CH2-), palmitoleic acid (- C(O)(CH2)7CHCH(CH2)5CH2-), vaccenic acid (-C(O)(CH2)9CHCH(CH2)5CH2-), paullinic acid
(-C(O)(CH2)IICHCH(CH2)5CH2-), oleic acid (-C(O)(CH2)7CHCH(CH2)7CH2-), elaidic acid
(-C(O)(CH2)7CHCH(CH2)7CH2-), gondoic acid (-C(O)(CH2)9CHCH(CH2)7CH2-), gadoleic acid (- C(O)(CH2)7CHCH(CH2)9CH2-), nervonic acid (- C(O)(CH2)i3CHCH(CH2)3CH2-), mead acid (- C(O)(CH2)3(CHCHCH2)3(CH2)6CH2-), myristoleic acid (-C(O)(CH2)7CHCH(CH2)3CH2-), and erucic acid (- C(O)(CH2)IICHCH(CH2)7CH2-).
In certain embodiments, in the compound of Formula II, Linker13 is selected from:
Figure imgf000192_0001
wherein:
R22 is independently at each occurrence selected from the group consisting of alkyl, - C(O)N-, -NC(O)-, -N-, -C(R21)-, -P(O)O-, -P(O)-, -P(O)(NR6R7)N-, alkenyl, haloalkyl, aryl, heterocycle, and heteroaryl, each of which is optionally substituted with 1, 2, 3, or 4 substituents independently selected from R21; and the remaining variables are as defined herein.
In certain embodiments, in the compound of Formula II, Linker0 is selected from:
Figure imgf000192_0002
wherein:
R32 is independently at each occurrence selected from the group consisting of alkyl, N+X-, -C-, alkenyl, haloalkyl, aryl, heterocycle, and heteroaryl, each of which is optionally substituted with 1, 2, 3, or 4 substituents independently selected from R21;
X- is an anionic group, for example Br- or Cl‘; and all other variables are as defined herein.
In certain embodiments, in the compound of Formula II, Linked is selected from:
Figure imgf000192_0003
wherein each heteroaryl, heterocycle, cycloalkyl, and aryl can optionally be substituted with 1, 2, 3, or 4 of any combination of halogen, alkyl, haloalkyl, and, heteroaryl, heterocycle, or cycloalkyl, as allowed by valence.
In certain embodiments, in the compound of Formula II, Linked is selected from:
Figure imgf000193_0001
wherein each heteroaryl, heterocycle, cycloalkyl, and and can optionally be substituted with 1, 2, 3, or 4 of any combination of halogen, alkyl, haloalkyl, aryl, heteroaryl, heterocycle, or cycloalkyl, as allowed by valence. In certain embodiments, in the compound of Formula II, Linker8 is selected from:
Figure imgf000193_0002
In certain embodiments, in the compound of Formula II, Linker8 is selected from:
Figure imgf000194_0001
In certain embodiments, in the compound of Formula II, Linker8, Linkerc, or Linker0 is selected from:
Figure imgf000195_0001
wherein tt is independently selected from 1, 2, or 3 and ss is 3 minus tt (3-tt).
In certain embodiments, in the compound of Formula II, Linker8, Linker13, or Linker0ted from:
Figure imgf000195_0002
wherein tt and ss are as defined herein.
In certain embodiments, in the compound of Formula II, Linker8, Linker13, or Linker0ted from:
Figure imgf000196_0001
Figure imgf000197_0001
Figure imgf000198_0001
Figure imgf000199_0001
wherein each heteroaryl, heterocycle, cycloalkyl, and aryl can optionally be substituted with 1, 2, 3, or 4 of any combination of halogen, alkyl, haloalkyl, aryl, heteroaryl, heterocycle, or cycloalkyl, as allowed by valence; and tt and ss are as defined herein.
In certain embodiments, in the compound of Formula II, Linker8, Linker13, or Linker0 is selected from:
Figure imgf000200_0001
Figure imgf000201_0001
wherein each heteroaryl, heterocycle, cycloalkyl, and aryl can optionally be substituted with 1, 23, or 4 of any combination of halogen, alkyl, haloalkyl, and, heteroaryl, heterocycle, or cycloalkyl, as allowed by valence: and tt and ss are as defined herein. In certain embodiments, in the compound of Formula II, LinkerB , LinkerC, or LinkerD is selected from:
Figure imgf000202_0001
wherein each heteroaryl and aryl can optionally be substituted with 1, 2, 3, or 4 of any combination of halogen, alkyl, haloalkyl, aryl, heteroaryl, heterocycle, or cycloalkyl, as allowed by valence; and tt and ss are as defined herein. In certain embodiments, in the compound of Formula II, LinkerA is selected from:
Figure imgf000202_0002
In certain embodiments, in the compound of Formula II, Linked is selected from:
Figure imgf000202_0003
In certain embodiments, in the compound of Formula II, Linked is selected from:
Figure imgf000203_0001
In certain embodiments, in the compound of Formula II, LinkerA is selected from:
Figure imgf000203_0002
In certain embodiments, in the compound of Formula II, LinkerB is selected from:
Figure imgf000203_0003
In certain embodiments, in the compound of Formula II, LinkerB is selected from:
Figure imgf000204_0001
In certain embodiments, in the compound of Formula II, LinkerB is selected from:
Figure imgf000204_0002
Figure imgf000205_0001
In certain embodiments, in the compound of Formula II, LinkerB is selected from:
Figure imgf000205_0002
Figure imgf000206_0001
In certain embodiments, in the compound of Formula II, LinkerC is selected from:
Figure imgf000206_0002
In certain embodiments, in the compound of Formula II, LinkerC is selected from:
Figure imgf000207_0001
In certain embodiments, in the compound of Formula II, LinkerC is selected from:
Figure imgf000207_0002
Figure imgf000208_0001
In certain embodiments, in the compound of Formula II, LinkerC is selected from:
Figure imgf000208_0002
In certain embodiments, in the compound of Formula II, LinkerC is selected from:
Figure imgf000209_0001
In certain embodiments, in the compound of Formula II, LinkerC is selected from:
Figure imgf000209_0002
In certain embodiments, in the compound of Formula II, LinkerC is selected from:
Figure imgf000210_0001
In certain embodiments, in the compound of Formula II, LinkerC is selected from:
Figure imgf000211_0001
In certain embodiments, in the compound of Formula II, LinkerD is selected from:
Figure imgf000212_0001
Figure imgf000213_0001
In certain embodiments, in the compound of Formula II, LinkerD is selected from:
Figure imgf000213_0002
In certain embodiments, in the compound of Formula II, LinkerD is selected from:
Figure imgf000214_0001
In certain embodiments, in the compound of Formula II, LinkerD is selected from:
Figure imgf000215_0001
In certain embodiments, in the compound of Formula II, LinkerD is selected from:
Figure imgf000215_0002
In certain embodiments, in the compound of Formula II, LinkerD is selected from:
Figure imgf000216_0001
In certain embodiments, in the compound of Formula II, the LinkerA is selected from
Figure imgf000217_0001
In certain embodiments, in the compound of Formula II, the LinkerA is selected from
Figure imgf000217_0002
In certain embodiments, in the compound of Formula II, the LinkerA is selected from
Figure imgf000217_0003
In certain embodiments, the LinkerA is selected from
Figure imgf000217_0004
Figure imgf000218_0001
wherein each is optionally substituted with 1, 2, 3, or 4 substituents substituent selected from R21.
In certain embodiments, in the compound of Formula II, Linked is selected from:
Figure imgf000218_0002
In certain embodiments, in the compound of Formula II, the LinkerA is selected from
Figure imgf000218_0003
In certain embodiments, in the compound of Formula II, the LinkerA is selected from
Figure imgf000219_0001
In certain embodiments, in the compound of Formula II, the LinkerA is selected from
Figure imgf000219_0002
In certain embodiments, in the compound of Formula II, the LinkerA is selected from
Figure imgf000219_0003
In certain embodiments, in the compound of Formula II, the LinkerA is selected from
Figure imgf000219_0004
In certain embodiments, in the compound of Formula II, the LinkerA is selected from In certain embodiments, in the compound of Formula II, the LinkerA is selected from
Figure imgf000220_0001
Figure imgf000221_0001
In certain embodiments, in the compound of Formula II, the LinkerA is selected from
Figure imgf000221_0002
Figure imgf000222_0001
In certain embodiments, in the compound of Formula II, the LinkerA is selected from
Figure imgf000222_0002
Figure imgf000223_0001
In certain embodiments, in the compound of Formula II, the LinkerA is selected from
Figure imgf000223_0002
In certain embodiments, in the compound of Formula II, the LinkerA is selected from
Figure imgf000223_0003
In certain embodiments, in the compound of Formula II, the LinkerA is selected from
Figure imgf000223_0004
Figure imgf000224_0001
In certain embodiments, in the compound of Formula II, the LinkerA is selected from
Figure imgf000224_0002
In certain embodiments, in the compound of Formula II, the LinkerA is selected from
Figure imgf000224_0003
In certain embodiments, in the compound of Formula II, the LinkerB is selected from
Figure imgf000225_0001
In certain embodiments, in the compound of Formula II, the LinkerB is selected from
Figure imgf000225_0002
In certain embodiments, in the compound of Formula II, the LinkerB is selected from
Figure imgf000225_0003
In certain embodiments, in the compound of Formula II, the LinkerB is selected from wherein each is optionally substituted with 1, 2, 3, or 4 substituents substituent selected from R21. In certain embodiments, in the compound of Formula II LinkerB is selected from:
Figure imgf000225_0004
In certain embodiments, in the compound of Formula II, the LinkerB is selected from:
Figure imgf000226_0001
In certain embodiments, in the compound of Formula II, the LinkerB is selected from:
Figure imgf000226_0002
In certain embodiments, in the compound of Formula II, the LinkerB is selected from:
Figure imgf000226_0003
In certain embodiments, in the compound of Formula II, the LinkerB is selected from:
Figure imgf000226_0004
In certain embodiments, in the compound of Formula II, the LinkerB is selected from:
Figure imgf000227_0001
In certain embodiments, in the compound of Formula II, the LinkerB is selected from:
Figure imgf000227_0002
In certain embodiments, in the compound of Formula II, the LinkerB is selected from:
Figure imgf000228_0001
In certain embodiments, in the compound of Formula II, LinkerB-LinkerA is selected from:
Figure imgf000228_0002
In certain embodiments, in the compound of Formula II, LinkerB-LinkerA is selected from:
Figure imgf000228_0003
In certain embodiments, in the compound of Formula II, the LinkerC is selected from:
Figure imgf000228_0004
In certain embodiments, in the compound of Formula II, the LinkerC is selected from:
Figure imgf000229_0001
In certain embodiments, in the compound of Formula II, the LinkerC is selected from:
Figure imgf000229_0002
In certain embodiments, in the compound of Formula II, the LinkerC is selected from:
Figure imgf000229_0003
In certain embodiments, in the compound of Formula II, the LinkerC is selected from:
Figure imgf000230_0001
In certain embodiments, in the compound of Formula II, the LinkerC is selected from:
Figure imgf000230_0002
In certain embodiments, in the compound of Formula II, the LinkerC is selected from:
Figure imgf000231_0001
Figure imgf000232_0001
Figure imgf000233_0001
In certain embodiments, in the compound of Formula II, the LinkerC is selected from: wherein each is optionally substituted with 1, 2, 3, or 4 substituents substituent selected from R21. In certain embodiments, in the compound of Formula II, the LinkerC is selected from:
Figure imgf000233_0002
In certain embodiments, in the compound of Formula II, the LinkerC is selected from:
Figure imgf000233_0003
In certain embodiments, in the compound of Formula II, the LinkerC is selected from:
Figure imgf000234_0001
In certain embodiments, in the compound of Formula II, the LinkerC is selected from:
Figure imgf000234_0002
In certain embodiments, in the compound of Formula II, the LinkerC is selected from:
Figure imgf000234_0003
In certain embodiments, in the compound of Formula II, the LinkerC is selected from:
Figure imgf000234_0004
In certain embodiments, in the compound of Formula II, the LinkerC is selected from:
Figure imgf000235_0001
In certain embodiments, in the compound of Formula II, the LinkerC is selected from:
Figure imgf000235_0002
In certain embodiments, in the compound of Formula II, LinkerC -(LinkerA)2 is selected from:
Figure imgf000235_0003
In certain embodiments, in the compound of Formula II, LinkerC -(LinkerA)2 is selected from:
Figure imgf000236_0001
In certain embodiments, in the compound of Formula II, LinkerC-(LinkerA)2 is selected from:
Figure imgf000236_0002
In certain embodiments, in the compound of Formula II, LinkerC-(LinkerA)2 is selected from:
Figure imgf000237_0001
In certain embodiments, in the compound of Formula II, LinkerD is selected from:
Figure imgf000238_0001
In certain embodiments, in the compound of Formula II, LinkerD is selected from:
Figure imgf000238_0002
wherein each is optionally substituted with 1, 2, 3, or 4 substituents are selected from R21. In certain embodiments, in the compound of Formula II, LinkerB -(LinkerA) is selected from
Figure imgf000239_0001
In certain embodiments, in the compound of Formula II, Linkerc-(LinkerA) is selected from
Figure imgf000239_0002
Figure imgf000240_0001
In certain embodiments, in the compound of Formula II, Linker°-(LinkerA) is selected from
Figure imgf000240_0002
In various embodiments, R4 is independently selected at each occurrence from hydrogen, heteroalkyl, alkyl, haloalkyl, arylalkyl, heteroarylalkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocycle, -OR6, -NR6R7, C(O)R3, S(O)R3, C(S)R3, and S(O)2R3.
In various embodiments, in the compound of Formula II, R5 is independently selected
Figure imgf000241_0001
from hydrogen, heteroalkyl, , C0-C6alkyl-cyano, alkyl, alkenyl, alkynyl, haloalkyl, F, Cl, Br, I, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocycle, heterocycloalkyl, haloalkoxy, -O-alkenyl, -O-alkynyl, C0-C6alkyl- OR6, C0-C6alkyl-SR6, C0- C6alkyl-NR6R7, C0-C6alkyl-C(O)R3, C0-C6alkyl-S(O)R3, C0-C6alkyl- C(S)R3, C0-C6alkyl- S(O)2R3, C0-C6alkyl-N(R8)-C(O)R3, C0-C6alkyl-N(R8)-S(O)R3, C0-C6alkyl- N(R8)-C(S)R3, C0-C6alkyl-N(R8)-S(O)2R3 C0-C6alkyl-O-C(O)R3, C0-C6alkyl-O-S(O)R3, C0- C6alkyl-O- C(S)R3, -N=S(O)(R3)2, C0-C6alkylN3, and C0-C6alkyl-O-S(O)2R3, each of which is optionally substituted with 1, 2, 3, or 4 substituents. In various embodiments, in the compound of Formula II, R6 and R7 are independently selected at each occurrence from hydrogen, heteroalkyl, alkyl, arylalkyl, heteroaryl alkyl, alkenyl, alkynyl, and, haloalkyl, heteroaryl, heterocycle, -alkyl-OR8, -alkyl-NR8R9, C(O)R3, S(O)R3, C(S)R3, and S(O)2R3. In various embodiments, in the compound of Formula II, R8 and R9 are independently selected at each occurrence from hydrogen, heteroalkyl, alkyl, arylalkyl, heteroarylalkyl, alkenyl, alkynyl, aryl, heteroaryl, and heterocycle. In various embodiments, the compound of Formula II has the structure of Formula II- A. In various embodiments, in the compound of Formula II-A, [MIFBM] and [IgGBM] are as defined herein. A compound of Formula II-A havin the structure:
Figure imgf000241_0002
wherein: [MIFBM/IgGBM] is a MIF or IgG Binding Moiety which binds respectively to circulating MIF or IgG in a subject, each of which is related to a disease state or condition and is to be removed by action of hepatocytes or other cells of the subject; [ASGPBM] is an asialoglycoprotein receptor binding moiety having the structure selected from
Figure imgf000242_0001
each [CON] is an optional connector chemical moiety which, when present, connects the [LIN] to [CPBM] or to [ASGPBM]; [LIN] is [LINKER] or [LINKER-2], each of which is a chemical moiety having a valency from 1 to 15, which covalently attaches to one or more [ASGPBM] or [CPBM] groups, optionally through a [CON], wherein the [LIN] optionally itself contains one or more [CON] groups; ZB is absent, (CH2)IM, C(O)-(CH2)IM-, or C(O)-(CH2)IM-NRM; RM is H or a C1-C3 alkyl group optionally substituted with one or two hydroxyl groups;
Figure imgf000243_0001
wherein RAM is H, C1-C4 alkyl optionally substituted with up to 3 halo groups and one or two hydroxyl groups, -(CH2)KCOOH, -(CH2)KC(O)O-(C1-C4 alkyl) optionally substituted with 1-3 halo groups, -O-C(O)-(C1-C4 alkyl) optionally substituted with 1-3 halo groups, - C(O)-(C1-C4 alkyl) optionally substituted with 1-3 halo groups, or -(CH2)K-NRN3RN4,or
Figure imgf000243_0002
wherein RTA is H, CN, NRN1RN2, -(CH2)KOH, -(CH2)KO(C1-C4 alkyl) optionally substituted with 1-3 halo groups, C1-C4 alkyl optionally substituted with 1-3 halo groups, - (CH2)KCOOH, -(CH2)KC(O)O-(C1-C4 alkyl) optionally substituted with 1-3 halo groups, -O- C(O)-(C1-C4 alkyl) optionally substituted with 1-3 halo groups, or -C(O)-(C1-C4 alkyl) optionally substituted with 1-3 halo groups, or RTA is a C3-C10 aryl or a three- to ten-membered heteroaryl group containing up to 5 heteroaryl atoms, each of the aryl or heteroaryl groups being optionally substituted with up to three CN, NRN1RN2, -(CH2)KOH, -(CH2)KO(C1-C4 alkyl) optionally substituted with 1-3 halo groups, C1-C3 alkyl optionally substituted with 1-3 halo groups or 1-2 hydroxy groups, -O-(C1-C3-alkyl) optionally substituted from 1-3 halo groups, -(CH2)KCOOH, - (CH2)KC(O)O-(C1-C4 alkyl) optionally substituted with 1-3 halo groups, O-C(O)-(C1-C4 alkyl) optionally substituted with 1-3 halo groups, or -(CH2)KC(O)-(C1-C4 alkyl) optionally substituted with 1-3 halo groups, or
Figure imgf000244_0001
Figure imgf000244_0002
optionally substituted with up to three C1-C3 alkyl groups which are optionally substituted with up to three halo groups; or
Figure imgf000244_0003
D. Other-Based ASGPR-Binding Moieties
In some embodiments, the ASGPR binding moieties can be any of the moieties described in: Reshitko, G. S., et al., “Synthesis and Evaluation of New Trivalent Ligands for Hepatocyte Targeting via the Asialoglycoprotein Receptor,” Bioconjugate Chem, doi: 10.1021/acs.bioconjchem.0c00202; Majouga, A. G., et al., “Identification of Novel Small- Molecule ASGP-R Ligands,” Current Drug Delivery, 2016, 13, 1303-1312, doi: 10.2174/1567201813666160719144651; Olshanova, A. S., et al., “Synthesis of a new betulinic acid glycoconjugate with N-acetyl-D-galactosamine for the targeted delivery to hepatocellular carcinoma cells,” Russian Chemical Bulletin, International Edition, Vol. 69, No. 1, pp. 158 — 163, January 2020; Yamansarov, E. Yu., et al., “New ASGPR-targeted ligands based on glycoconjugated natural triterpenoids,” Russian Chemical Bulletin, International Edition, Vol. 68, No. 12, pp. 2331 — 2338, December 2019; Congdon, M. D., et al., “Enhanced Binding and Reduced Immunogenicity of Glycoconjugates Prepared via Solid-State Photoactivation of Aliphatic Diazirine Carbohydrates,” Bioconjugate Chem, doi: 10.1021/acs.bioconjchem.0c00555; and Dhawan, V., et al., “Polysaccharide conjugates surpass monosaccharide ligands in hepatospecific targeting - Synthesis and comparative in silico and in vitro assessment,” Carbohydrate Research 509 (2021) 108417, doi:
10.1016/j. carres.2021.108417. The following ASGPR binding moieties are illustrative and not intended to be limiting.
1. GalNAc-Tyrosine Based Moieties
In some embodiments, the ASGPR binding moiety can be a moiety having the structure of Ml, M2, M3, or M4, or a combination thereof. In the structures of Ml, M2, M3, and M4, X is independently at each occurrence O, NH, or S. In various embodiments, compounds of Formula I or Formula II can have one, two, or three ASGPR binding moieties with the structure of Ml, M2, M3, or M4.
Figure imgf000245_0001
M3 M4.
In various embodiments, ASGPR binding moieties Ml to M4 can be conjugated to any suitable [CON], [Linker], or [Linker-2] as described herein and in Congdon, M. D., et al., “Enhanced Binding and Reduced Immunogenicity of Glycoconjugates Prepared via Solid- State Photoactivation of Aliphatic Diazirine Carbohydrates,” Bioconjugate Chem. doi: 10.1021/acs.bioconjchem.0c00555.
2. Trivalent Triazole-Based Moieties
In some embodiments, the ASGPR binding moiety can be a moiety having the structure of M5 :
Figure imgf000246_0001
, M5. In the structures M5, each R is independently at each occurrence R1 or R2,
Figure imgf000246_0002
. In various embodiments, compounds of Formula I or Formula II contain an ASGPR binding moiety with the structure of M5. In various embodiments, each R in M5 is R1. In various embodiments, each R in M5 is R2. In various embodiments, ASGPR binding moiety M5 can be conjugated/bonded to any suitable [CON], [Linker], or [Linker-2] as described herein and in Reshitko, G. S., et al., “Synthesis and Evaluation of New Trivalent Ligands for Hepatocyte Targeting via the Asialoglycoprotein Receptor,” Bioconjugate Chem, doi: 10.1021/acs.bioconjchem.0c00202. 3. Galactose- and Agarose-derived Behenic Acid Ester Moieties In various embodiments, the ASGPR binding moiety can be the galactose behenic acid ester-derived moiety M7:
Figure imgf000246_0003
In the structure M7, Y is OH or NHAc. In various embodiments, the ASGPR binding moiety can be the agarose behenic acid ester-derived moiety M8:
Figure imgf000247_0001
In various embodiments, ASGPR binding moieties M7 and M8 can be conjugated to any suitable [CON], [Linker], or [Linker-2] as described herein and in Dhawan, V., et al., “Polysaccharide conjugates surpass monosaccharide ligands in hepatospecific targeting - Synthesis and comparative in silico and in vitro assessment,” Carbohydrate Research 509 (2021) 108417, doi: 10.1016/j.carres.202L 108417.
4. Other Small Molecule ASGPR Binding Moieties In various embodiments, the ASGPR binding moiety can be any of the compounds 2-
18 below:
Figure imgf000247_0002
Figure imgf000248_0001
14 15 16
Figure imgf000248_0002
17 18 In various embodiments, in compounds 15 and 16, R is CILOAc, COOH, or CH2OH.
Compounds 2-18 can be conjugated/bonded to any suitable [CON], [Linker], or [Linker-2] as described herein and in Majouga, A. G., et al., “Identification of Novel Small-Molecule ASGP-R Ligands,” Current Drug Delivery, 2016, 13, 1303-1312, doi: 10.2174/1567201813666160719144651; Olshanova, A. S., et al., “Synthesis of a new betulinic acid glycoconjugate with N-acetyl-D-galactosamine for the targeted delivery to hepatocellular carcinoma cells,” Russian Chemical Bulletin, International Edition, Vol. 69, No. 1, pp. 158 — 163, January 2020; Yamansarov, E. Yu., et al., “New ASGPR-targeted ligands based on glycoconjugated natural triterpenoids,” Russian Chemical Bulletin, International Edition, Vol. 68, No. 12, pp. 2331 — 2338, December 2019. Compounds 2-18 can be attached through any suitable reactive group contained therein. Without limitation, compounds 2-13 can be attached to a CON], [Linker], or [Linker-2] through or by reaction with at least one OH, NH, vinyl, alkynyl, amide, acid, ester, ketone, or aromatic halogen contained in compounds 2-18. Suitable reaction modes for attaching compounds 2-18 to a [CON], [Linker], or [Linker-2] as described herein include, but are not limited to, substitution (e.g. alkylation of OH or NH groups), esterification (forming an ester), amidation (forming an amide), transesterification (exchanging one ester for another), transamidation (exchanging one amide for another), azide-alkyne cycloaddition, and other reactions capable of forming C-C, N-C, or O-C bonds with vinyl and alkynyl groups such as cycloadditions, aminations, oxidations, alkylations, rearrangement reactions (e.g. Claisen, Cope, etc.), and the like.
The term “pharmaceutically acceptable salt” or “salt” is used throughout the specification to describe a salt form of one or more of the compositions herein which are presented to increase the solubility of the compound in saline for parenteral delivery or in the gastric juices of the patient’s gastrointestinal tract in order to promote dissolution and the bioavailability of the compounds. Pharmaceutically acceptable salts include those derived from pharmaceutically acceptable inorganic or organic bases and acids. Suitable salts include those derived from alkali metals such as potassium and sodium, alkaline earth metals such as calcium, magnesium and ammonium salts, among numerous other acids well known in the pharmaceutical art. In some embodiments, sodium and potassium salts are used as neutralization salts of carboxylic acids and free acid phosphate containing compositions according to the present disclosure. The term “salt” shall mean any salt consistent with the use of the compounds according to the present disclosure. In the case where the compounds are used in pharmaceutical indications, including the treatment of prostate cancer, including metastatic prostate cancer, the term “salt” shall mean a pharmaceutically acceptable salt, consistent with the use of the compounds as pharmaceutical agents.
The term “coadministration” shall mean that at least two compounds or compositions are administered to the patient at the same time, such that effective amounts or concentrations of each of the two or more compounds may be found in the patient at a given point in time. Although compounds according to the present disclosure may be co-administered to a patient at the same time, the term embraces both administration of two or more agents at the same time or at different times, provided that effective concentrations of all coadministered compounds or compositions are found in the subject at a given time. Chimeric antibodyrecruiting compounds according to the present disclosure may be administered with one or more additional anti-cancer agents or other agents which are used to treat or ameliorate the symptoms of cancer, especially prostate cancer, including metastatic prostate cancer.
The term “anticancer agent” or “additional anticancer agent” refers to a compound other than the chimeric compounds according to the present disclosure which may be used in combination with a compound according to the present disclosure for the treatment of cancer. Exemplary anticancer agents which may be coadministered in combination with one or more chimeric compounds according to the present disclosure include, for example, antimetabolites, inhibitors of topoisomerase I and II, alkylating agents and microtubule inhibitors (e.g., taxol), among others. Exemplary anticancer compounds for use in the present disclosure may include everolimus, trabectedin, abraxane, TLK 286, AV-299, DN-101 , pazopanib, GSK690693, RTA 744, ON O91O.Na, AZD 6244 (ARRY-142886), AMN-107, TKI-258, GSK461364, AZD 1152, enzastaurin, vandetanib, ARQ-197, MK-0457, MLN8054, PHA-739358, R-763, AT-9263, a FLT-3 inhibitor, a VEGFR inhibitor, an EGFR TK inhibitor, an aurora kinase inhibitor, a PIK-1 modulator, a Bcl-2 inhibitor, an HD AC inhbitor, a c-MET inhibitor, a PARP inhibitor, a Cdk inhibitor, an EGFR TK inhibitor, an IGFR-TK inhibitor, an anti-HGF antibody, a PI3 kinase inhibitors, an AKT inhibitor, a JAK/STAT inhibitor, a checkpoint- 1 or 2 inhibitor, a focal adhesion kinase inhibitor, a Map kinase kinase (mek) inhibitor, a VEGF trap antibody, pemetrexed, erlotinib, dasatanib, nilotinib, decatanib, panitumumab, amrubicin, oregovomab, Lep-etu, nolatrexed, azd2171, batabulin, ofatumumab (Arzerra), zanolimumab, edotecarin, tetrandrine, rubitecan, tesmilifene, oblimersen, ticilimumab, ipilimumab, gossypol, Bio 111 , 131-I-TM-601 , ALT-110, BIO 140, CC 8490, cilengitide, gimatecan, IL13-PE38QQR, INO 1001 , IPdRi KRX-0402, lucanthone, LY 317615, neuradiab, vitespan, Rta 744, Sdx 102, talampanel, atrasentan, Xr 311 , romidepsin, ADS- 100380, sunitinib, 5-fluorouracil, vorinostat, etoposide, gemcitabine, doxorubicin, irinotecan, liposomal doxorubicin, 5'-deoxy-5-fluorouridine, vincristine, temozolomide, ZK- 304709, seliciclib; PD0325901 , AZD-6244, capecitabine, L-Glutamic acid, N -[4-[2-(2- amino-4,7-dihydro-4-oxo-l H - pyrrolo[2,3- d ]pyrimidin-5-yl)ethyl]benzoyl]-, disodium salt, heptahydrate, camptothecin, PEG-labeled irinotecan, tamoxifen, toremifene citrate, anastrazole, exemestane, letrozole, DES(diethylstilbestrol), estradiol, estrogen, conjugated estrogen, bevacizumab, IMC-1C11 , CHIR-258,); 3-[5-(methylsulfonylpiperadinemethyl)- indolylj -quinolone, vatalanib, AG-013736, AVE-0005, the acetate salt of [D- Ser(Bu t ) 6 ,Azgly 10 ] (pyro-Glu-His-Trp-Ser-Tyr-D-Ser(Bu t )-Leu- Arg-Pro- Azgly-NH 2 acetate [C59H84NisOi4 -(C2H4O2)x where x = 1 to 2.4], goserelin acetate, leuprolide acetate, triptorelin pamoate, medroxyprogesterone acetate, hydroxyprogesterone caproate, megestrol acetate, raloxifene, bicalutamide, flutamide, nilutamide, megestrol acetate, CP-724714; TAK-
165, HKI-272, erlotinib, lapatanib, canertinib, ABX-EGF antibody, erbitux, EKB-569, PKL
166, GW-572016, lonafamib, BMS-214662, tipifarnib; amifostine, NVP-LAQ824, suberoyl analide hydroxamic acid, valproic acid, trichostatin A, FK-228, SU11248, sorafenib, KRN951 , aminoglutethimide, arnsacrine, anagrelide, L-asparaginase, Bacillus Calmette- Guerin (BCG) vaccine, bleomycin, buserelin, busulfan, carboplatin, carmustine, chlorambucil, cisplatin, cladribine, clodronate, cyproterone, cytarabine, dacarbazine, dactinomycin, daunorubicin, diethylstilbestrol, epirubicin, fludarabine, fludrocortisone, fluoxymesterone, flutamide, gemcitabine, gleevac, hydroxyurea, idarubicin, ifosfamide, imatinib, leuprolide, levamisole, lomustine, mechlorethamine, melphalan, 6-mercaptopurine, mesna, methotrexate, mitomycin, mitotane, mitoxantrone, nilutamide, octreotide, oxaliplatin, pamidronate, pentostatin, plicamycin, porfimer, procarbazine, raltitrexed, rituximab, streptozocin, teniposide, testosterone, thalidomide, thioguanine, thiotepa, tretinoin, vindesine, 13-cis-retinoic acid, phenylalanine mustard, uracil mustard, estramustine, altretamine, floxuridine, 5-deooxyuridine, cytosine arabinoside, 6-mecaptopurine, deoxycoformycin, calcitriol, valrubicin, mithramycin, vinblastine, vinorelbine, topotecan, razoxin, marimastat, COL-3, neovastat, BMS-275291 , squalamine, endostatin, SU5416, SU6668, EMD121974, interleukin- 12, IM862, angiostatin, vitaxin, droloxifene, idoxyfene, spironolactone, finasteride, cimitidine, trastuzumab, denileukin diftitox,gefitinib, bortezimib, paclitaxel, irinotecan, topotecan, doxorubicin, docetaxel, vinorelbine, bevacizumab (monoclonal antibody) and erbitux, cremophor-free paclitaxel, epithilone B, BMS- 247550, BMS-310705, droloxifene, 4-hydroxytam oxifen, pipendoxifene, ERA- 923, arzoxifene, fulvestrant, acolbifene, lasofoxifene, idoxifene, TSE-424, HMR- 3339, ZK 186619, PTK787/ZK 222584, VX-745, PD 184352, rapamycin, 40-O-(2-hydroxyethyl)-rapamycin, temsirolimus, AP- 23573, RAD001 , ABT-578, BC-210, LY294002, LY292223, LY292696, LY293684, LY293646, wortmannin, ZM336372, L-779,450, PEG-filgrastim, darbepoetin, erythropoietin, granulocyte colony-stimulating factor, zolendronate, prednisone, cetuximab, granulocyte macrophage colony-stimulating factor, histrelin, pegylated interferon alfa-2a, interferon alfa- 2a, pegylated interferon alfa-2b, interferon alfa-2b, azacitidine, PEG-L-asparaginase, lenalidomide, gemtuzumab, hydrocortisone, interleukin- 11 , dexrazoxane, alemtuzumab, all- transretinoic acid, ketoconazole, interleukin-2, megestrol, immune globulin, nitrogen mustard, methylprednisolone, ibritgumomab tiuxetan, androgens, decitabine, hexamethylmelamine, bexarotene, tositumomab, arsenic trioxide, cortisone, editronate, mitotane, cyclosporine, liposomal daunorubicin, Edwina-asparaginase, strontium 89, casopitant, netupitant, an NK-1 receptor antagonists, palonosetron, aprepitant, diphenhydramine, hydroxyzine, metoclopramide, lorazepam, alprazolam, haloperidol, droperidol, dronabinol, dexamethasone, methylprednisolone, prochlorperazine, granisetron, ondansetron, dolasetron, tropisetron, pegfilgrastim, erythropoietin, epoetin alfa and darbepoetin alfa, vemurafenib among others, including immunotherapy agents such as IDO inhibitors (an inhibitor of indoleamine 2,3-dioxygenase (IDO) pathway) such as Indoximod (NLG-8187), Navoximod (GDC-0919) and NLG802, PDL1 inhibitors (an inhibitor of programmed death-ligand 1) including, for example, nivolumab, durvalumab and atezolizumab, PD1 inhibitors such as pembrolizumab (Merck) and CTLA-4 inhibitors (an inhibitor of cytotoxic T-lymphocyte associated protein 4/cluster of differentiation 152), including ipilimumab and tremelimumab, among others.
In addition to anticancer agents, a number of other agents may be coadministered with chimeric compounds according to the present disclosure in the treatment of cancer. These include active agents, minerals, vitamins and nutritional supplements which have shown some efficacy in inhibiting cancer tissue or its growth or are otherwise useful in the treatment of cancer. For example, one or more of dietary selenium, vitamin E, lycopene, soy foods, curcumin (turmeric), vitamin D, green tea, omega-3 fatty acids and phytoestrogens, including beta-sitosterol, may be utilized in combination with the present compounds to treat cancer.
Without not being limited by way of theory, compounds according to the present disclosure which contain a MIF binding moiety (MIFBM) or antibody binding moiety and ASGPR binding moiety selectively bind to liver cells and through that binding, facilitate the introduction of the MIFBM group into hepatocytes which bind the ASGPRBM selectively, where, the MIF protein, inside the hepatocyte is degraded and removed from circulation. Thus, compounds according to the present disclosure both bind to MIF proteins and remove the MIF proteins from circulation resulting in a dual action which is particularly effective for treating disease states and conditions.
Pharmaceutical compositions comprising combinations of an effective amount of at least one compound disclosed herein, often a bi-functional chimeric compound (containing at least one MIFBM group or antibody binding moiety and at least one ASGPRBM) according to the present disclosure, and one or more of the compounds as otherwise described herein, all in effective amounts, in combination with a pharmaceutically effective amount of a carrier, additive or excipient, represents a further aspect of the present disclosure. These may be used in combination with at least one additional, optional anticancer agent as otherwise disclosed herein.
The compositions of the present disclosure may be formulated in a conventional manner using one or more pharmaceutically acceptable carriers and may also be administered in controlled-release formulations. Pharmaceutically acceptable carriers that may be used in these pharmaceutical compositions include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as prolamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene- polyoxypropylene-block polymers, polyethylene glycol and wool fat.
In some embodiments, the compositions of the present disclosure are administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir, among others. The term "parenteral" as used herein includes subcutaneous, intravenous, intramuscular, intra-articular, intra-synovial, intrastemal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. In some embodiments, the compositions are administered orally (including via intubation through the mouth or nose into the stomach), intraperitoneally or intravenously.
Sterile injectable forms of the compositions of this disclosure may be aqueous or oleaginous suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a nontoxic parenterally-acceptable diluent or solvent, for example as a solution in 1, 3 -butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono- or di-glycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant, such as Ph. Helv or similar alcohol.
The pharmaceutical compositions of this disclosure may be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, aqueous suspensions or solutions. In the case of tablets for oral use, carriers which are commonly used include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried corn starch. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring or coloring agents may also be added.
Alternatively, the pharmaceutical compositions of this disclosure may be administered in the form of suppositories for rectal administration. These can be prepared by mixing the agent with a suitable non-irritating excipient which is solid at room temperature but liquid at rectal temperature and therefore will melt in the rectum to release the drug. Such materials include cocoa butter, beeswax and polyethylene glycols.
The pharmaceutical compositions of this disclosure may also be administered topically, especially to treat skin cancers, psoriasis or other diseases which occur in or on the skin. Suitable topical formulations are readily prepared for each of these areas or organs. Topical application for the lower intestinal tract can be effected in a rectal suppository formulation (see above) or in a suitable enema formulation. Topically-acceptable transdermal patches may also be used.
For topical applications, the pharmaceutical compositions may be formulated in a suitable ointment containing the active component suspended or dissolved in one or more carriers. Carriers for topical administration of the compounds of this disclosure include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water.
Alternatively, the pharmaceutical compositions can be formulated in a suitable lotion or cream containing the active components suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2- octyl dodecanol, benzyl alcohol and water.
For ophthalmic use, the pharmaceutical compositions may be formulated as micronized suspensions in isotonic, pH adjusted sterile saline, or as solutions in isotonic, pH adjusted sterile saline, either with our without a preservative such as benzylalkonium chloride. Alternatively, for ophthalmic uses, the pharmaceutical compositions may be formulated in an ointment such as petrolatum.
The pharmaceutical compositions of this disclosure may also be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other conventional solubilizing or dispersing agents.
The amount of compound in a pharmaceutical composition of the instant disclosure that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host and disease treated, the particular mode of administration. In some embodiments, the compositions are formulated to contain between about 0.05 milligram to about 1.5 grams, from 0.1 milligram to 1 gram, 0.5 milligrm to 750 milligrams, more often about 1 milligram to about 600 milligrams, and even more often about 10 milligrams to about 500 milligrams of active ingredient, alone or in combination with at least one additional compound which may be used to treat cancer, prostate cancer or metastatic prostate cancer or a secondary effect or condition thereof.
Methods of treating patients or subjects in need for a particular disease state or condition as otherwise described herein, especially cancer, comprise administration of an effective amount of a pharmaceutical composition comprising therapeutic amounts of one or more of the novel compounds described herein and optionally at least one additional bioactive (e.g., anti-cancer, anti-inflammatory) agent. The amount of active ingredient(s) used in the methods of treatment of the instant disclosure that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated, the particular mode of administration. For example, the compositions could be formulated so that a therapeutically effective dose of between about 0.01, 0.1, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90 or 100 mg/kg of patient/day or in some embodiments, greater than 100, 110, 120, 130, 140, 150, 160, 170, 180, 190 or 200 mg/kg of the novel compounds can be administered to a patient receiving these compositions.
It should also be understood that a specific dosage and treatment regimen for any particular patient will depend upon a variety of factors, including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the judgment of the treating physician and the severity of the particular disease or condition being treated.
A patient or subject (e.g. a human) suffering from an autoimmune disease, an inflammatory disease or cancer can be treated by administering to the patient (subject) an effective amount of a chimeric/bi-functional compound according to the present disclosure including pharmaceutically acceptable salts, solvates or polymorphs, thereof optionally in a pharmaceutically acceptable carrier or diluent, either alone, or in combination with other known pharmaceutical agents, exemplary agents which can assist in treating autoimmune and/or inflammatory diseases or cancer, including metastatic cancer or recurrent cancer or ameliorating the secondary effects and/or symptoms assocated with these disease states and/or conditions. This treatment can also be administered in conjunction with other conventional therapies, such as radiation treatment or surgery for cancer.
The present compounds, alone or in combination with other agents as described herein, can be administered by any appropriate route, for example, orally, parenterally, intravenously, intradermally, subcutaneously, or topically, in liquid, cream, gel, or solid form, or by aerosol form.
The active compound is included in the pharmaceutically acceptable carrier or diluent in an amount sufficient to deliver to a patient a therapeutically effective amount for the desired indication, without causing serious toxic effects in the patient treated. An exemplary dose of the active compound for all of the herein-mentioned conditions is in the range from about 10 ng/kg to 300 mg/kg, such as 0.1 to 100 mg/kg per day, such as 0.5 to about 25 mg per kilogram body weight of the recipient/patient per day. Some exemplary dosages will range from about 0.01-3% wt/wt in a suitable carrier.
The compound is conveniently administered in any suitable unit dosage form, including but not limited to one containing less than Img, 1 mg to 3000 mg, such as 5 to 500 mg of active ingredient per unit dosage form. An oral dosage of about 25-500 mg is often convenient.
The active ingredient is sometimes administered to achieve peak plasma concentrations of the active compound of about 0.00001-30 mM, such as about 0.1-30 pM. This may be achieved, for example, by the intravenous injection of a solution or formulation of the active ingredient, optionally in saline, or an aqueous medium or administered as a bolus of the active ingredient. Oral administration is also appropriate to generate effective plasma concentrations of active agent.
The concentration of active compound in the drug composition will depend on absorption, distribution, inactivation, and excretion rates of the drug as well as other factors known to those of skill in the art. It is to be noted that dosage values will also vary with the severity of the condition to be alleviated. It is to be further understood that for any particular subject, specific dosage regimens should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions, and that the concentration ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed composition. The active ingredient may be administered at once, or may be divided into a number of smaller doses to be administered at varying intervals of time.
Oral compositions will generally include an inert diluent or an edible carrier. They may be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the active compound or its prodrug derivative can be incorporated with excipients and used in the form of tablets, troches, or capsules. Pharmaceutically compatible binding agents, and/or adjuvant materials can be included as part of the composition.
The tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a dispersing agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterotes; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring. When the dosage unit form is a capsule, it can contain, in addition to material of the above type, a liquid carrier such as a fatty oil. In addition, dosage unit forms can contain various other materials which modify the physical form of the dosage unit, for example, coatings of sugar, shellac, or enteric agents.
The active compound or pharmaceutically acceptable salt thereof can be administered as a component of an elixir, suspension, syrup, wafer, chewing gum or the like. A syrup may contain, in addition to the active compounds, sucrose as a sweetening agent and certain preservatives, dyes and colorings and flavors
The active compound or pharmaceutically acceptable salts thereof can also be mixed with other active materials that do not impair the desired action, or with materials that supplement the desired action, such as other anticancer agents, anti-inflammatory agents, immunosuppressants, antibiotics, antifungals, or antiviral compounds. In certain aspects of the disclosure, one or more chimeric/bi-functional MIF binding compound according to the present disclosure is coadministered with another anticancer agent and/or another bioactive agent, as otherwise described herein.
Solutions or suspensions used for parenteral, intradermal, subcutaneous, or topical application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. The parental preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.
If administered intravenously, carriers sometimes are physiological saline or phosphate buffered saline (PBS).
In certain embodiments, the active compounds are prepared with carriers that will protect the compound against rapid elimination from the body, such as a controlled and/or sustained release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Methods for preparation of such formulations will be apparent to those skilled in the art.
Liposomal suspensions or cholestosomes may also be pharmaceutically acceptable carriers. These may be prepared according to methods known to those skilled in the art, for example, as described in U.S. Pat. No. 4,522,811 (which is incorporated herein by reference in its entirety). For example, liposome formulations may be prepared by dissolving appropriate lipid(s) (such as stearoyl phosphatidyl ethanolamine, stearoyl phosphatidyl choline, arachadoyl phosphatidyl choline, and cholesterol) in an inorganic solvent that is then evaporated, leaving behind a thin film of dried lipid on the surface of the container. An aqueous solution of the active compound are then introduced into the container. The container is then swirled by hand to free lipid material from the sides of the container and to disperse lipid aggregates, thereby forming the liposomal suspension.
Chemical Synthesis
FIGURES 1, 7, and 13 attached hereto identifies exemplary compounds according to the present disclosure which exhibit activity in binding to and reducing and/or eliminating unwanted circulating proteins for therapeutic and/or diagnostic purposes. These compounds are based upon an MIF, anti-DNP IgG, or IgG binding moiety to which is covalently attached an ASPGR group such as GN3 or AcF3-3 group through a linker which contains from 1 to 100 ethylene glycol groups, more often from 1 to 15 ethylene glycol groups, from 1 to 10 ethylene glycol groups, often from 2 to 10 ethylene glycol groups which are optionally attached through a [CON] group, such as a 1,2,3-triazole or other [CON] group as described herein.
FIGURE 16 shows the synthesis of azide/amide carboxylic end capped PEG linker intermediates which may be condensed onto an alkynyl precursor (e.g. NVS alkyne precursor of Figure 5) to provide carboxylic acid capped intermediate which can be used to provide bifunctional molecules.
FIGURE 17 describes a general method for conversion of PEG molecules into hydroxyl azides. The PEG compound is tosylated (TsCl, DCM, in the presence of base) at reduced temperature and further reacted with sodium azide at elevated temperature in a non- nucleophilic solvent. The final azidoalcohol is used in subsequent figures. FIGURE 18 describes the synthesis of a mesylated azide from a starting PEG molecule employing the same synthetic steps to reach the intermediate azido alcohol. This is then treated with MsCl in pyridine to afford the final compound.
FIGURE 19 shows the synthesis of the GalNAc ASGPR ligand linked through PEG to a terminating amine. Pentaacetyl galactosamine is reacted with TMSOTf at elevated temperature in DCE to produce a bicyclic intermediate, which is then reacted with an azido alcohol to give an azide intermediate (TMSOTf, DCE). This molecule is then subjected to a Staudinger reduction to give an amine which is used in subsequent figures.
FIGURE 20 shows the synthesis of a higher affinity bicylic ASGPR ligand.
Galactose pentaacetate is treated with HBr/AcOH to give the brominated intermediate, which is treated with Zn and CuSO4 (water/ AcOH) to give the galactal. This is treated with ammonium cerium nitrate and sodium azide at reduced temperature (MeCN) to give the disubstituted intermediate compound. This is then treated with strong base (NaOMe/MeOH) to give the triol azide intermediate. This compound is silylated completely (TMSCl/pyr) then the primary alcohol is deprotected (potassium carbonate, MeOH, lowered temperature) and oxidized (Dess-Martin Periodinane, DCM). Treatment with strong base (NaOEt/HOEt) and paraformaldehyde gives the tetraol intermediate, which is cyclized in strong acid (H2SO4/water) to give the bicyclic azide ligand.
FIGURE 21 shows the synthesis of a trifluoro-acetate derivative of the bicyclic ASGPR ligand. The triol azide is reduced (Pd/C, MeOH) to give the intermediate amine, which is then peracylated with trifuloroacetic anhydride. The esters are hydrolyzed with strong base (NaOMe/HOMe) to give the intermediate amide, which is protected using dimethoxypropane in the presence of camphorsulfonic acid in DMF at elevated temperature. This is then reacted a mesylated azdio alcohol in the presence of strong base (NaH/DMF) to give an intermediate azide that is reduced (Lindlar’s catalyst, MeOH) to give the final amine.
FIGURE 22 shows the synthesis of the MIF -targeting linker to a monovalent linker, which is synthesized through analagous methods as described in a previous figure. The boc- protected methyl ester is deprotected with TFA in DCM, then coupled to the MIF-targeting carboxylic acid (HBTU, DIPEA, DMF). Subsequent hydrolysis with strong base (NaOH/dioxane/H2O) gives the MIF-targeting carboxylic acid.
FIGURE 23 shows the synthesis of the di-carboxylic acid MIF targeting motif, which is synthesized as described in previous figures.
FIGURE 24 shows the synthesis of a tris base-derived trivalent linker. Tris base is treated with di-t-butyl dicarbonate in the presence of base to give the boc protected triol, which is then reacted with acrylonitrile in the presence of base (dioxane/H2O) to give a trinitrile intermediate. This is then converted to the methyl ester through treatment with strong acid in methanol. The amine is then reacted with Cbz-glycine through a DCC- mediated amide formation, and deprotected to give a tricarboxylic acid that is used in subsequent figures.
FIGURE 25 describes the synthesis of an ASGPR-targeting moiety employing three GalNAc ASGPR ligands. The tricarboxylic acid is reacted with amine-terminated protected GalNAc (amide bond formation in the presence of HBTU and DIPEA), then deprotected by reduction (Pd/C, solvent) and treatment with strong base (NaOMe/MeOH).
FIGURE 26 shows the synthesis of the tri-carboxylic acid MIF targeting motif, which is synthesized as described in previous figures.
FIGURE 27 shows the synthesis of the MIF NVS alkyne precursor which can be reacted with an azido reactant containing a carboxylic acid (as set forth in subsequent figures) to provide MIF-NVS-carboxylic acid capped reactants to produce the bifunctional compounds.
FIGURE 28 shows the synthesis of the MIF-targeting moiety terminating in a carboxylic acid. 2-chloroquinolin-6-ol is reacted with ethyl 4-bromobutanoate in the presence of base (DMF, elevated temperature) to give an aryl chloride that then undergoes Sonogashira coupling at evelvated temperature with ethynyltrimethylsilane. The intermediate silylated compound is deprotected with TBAF (DCM/THF). A click reaction then forms a triazole between the alkyne intermediate and in situ synthesized 4-azido-2-fluorophenol to give an ethyl ester intermediate that is hydrolyzed with strong base (NaOH/dioxane) to give the carboxylic acid that is used in subsequent figures.
FIGURE 29 describes the synthesis of the bifunctional molecule MIF-NVS-PEGn- GN3 through HBTU-mediated coupling in DMF of the ASPGR-targeting amine and the MIF targeting carboxylic acid prepared by first forming the MIF-targeting carboxylic acid by condensing the reactant azido PEG-carboxylic acid onto the MIF moiety containing a alkyne terminated PEG group.
FIGURE 30 describes the synthesis of bifunctional molecules MIF-GN3 and MIF- PEGn-GN3 through HATU-mediated coupling (DMF, DIPEA) of ASGPR-targeting amine and MIF-targeting carboxylic acid.
FIGURE 31 describes the synthesis of the bifunctional molecule targeting MIF and ASGPR, containing one bicyclic ASGPR AcF3 ligands. MIF-binding mono-carboxylic acid is treated with HBTU, DIPEA, the amine terminated ligand, and DMF to give the amide, which is then deprotected with IM HC1 to give the final compound.
FIGURE 32 describes the synthesis of the bifunctional molecule targeting MIF and ASGPR, containing two bicyclic ASGPr ligands. It is synthesized as described above.
FIGURE 33 describes the synthesis of the bifunctional molecule targeting MIF and ASGPR, containing three bicyclic ASGPr ligands. It is synthesized as described above.
FIGURE 34 shows the synthesis of DNP-GN3. 2,4-dinitro chlorobenzene was treated with an amino carboxylic acid in the presence of weak base to give the di-nitro analine carboxylic acid intermediate. Further steps were carried out as described for previous molecules.
FIGURE 35 shows the synthesis of DNP-AcF3-3, which was carried out with methods analogous previous compounds.
FIGURE 36 shows the synthetic scheme used to obtain IBA-GN3. Pentaethylene glycol was treated with tosyl chloride in the presence of base to give the mono-tosylated alcohol, which was then treated with sodium azide at elevated temperature to give the azidoalcohol. This compound was then oxidized using Jones reagent, then reduced with Palladium on carbon under hydrogen atmosphere to give a carboxylic acid-amine. Separately, indole butyric acid was treated with N-hydroxysuccinimide, EDC, and DIPEA to give the NHS-ester indole, which was then reacted with the above carboxylic acid-amine. The product was again reacted with N-hydroxysuccinimide, EDC, and DIPEA to give a NHS ester. This NHS ester was reacted with NH2-GN3, which was prepared as described previously. The subsequent amide was deprotected with NaOMe in MeOH to give compound IBA-GN3.
FIGURE 37 shows the synthesis of triazine-GN3. Cyanuric chloride was treated with (4-(methoxycarbonyl)phenyl)methanaminium in THF and diisopropylethlamine at -78° C to give the mono-substituted product. This was then treated with cyclohexylmethanamine at room temperature to afford the second substitution. The final substitution was accomplished under elevated temperature with (lS,2S,4R)-bicyclo[2.2.1]heptan-2-amine to give the tri substituted triazine. Deprotection with lithium hydroxide followed by amide coupling with a monoprotected diamine gave the Boc-protected derivative, which was deprotected and reacted with glutaric anhydride to give a carboxylic acid that was converted to an NHS ester using standard coupling conditions. This was reacted with NH2-GN3 to give the final product.
FIGURE 38 shows the synthetic scheme used to access FcIII-GN3. The hexynyl peptide was prepared using standard solid phase peptide synthesis techniques. The peptide was removed from Rink resin using Reagent L, then oxidized using ammonium bicarbonate buffer (pH8-9) in MeOH under air to give the cyclic peptide. The peptide was reacted with GN3-azide, which was described previously, to give the product triazole FcIII-GN3.
FIGURE 39 shows the synthetic scheme used to access FcIII-4c-GN3, which was accomplished using methods described above.
FIGURES 40-43 describe the synthesis of bifunctional molecules targeting MIF and ASGPr, containing three bicyclic ASGPR ligands with different substitutions on the 2-amine of the sugar. They are synthesized through analagous methods described above as set forth in the attached figures.
FIGURE 44 shows the synthesis of compound MIF-18-3. Tri-acyl galactal was deprotected with ammonia in methanol, then tri-benzyl protected with benzylbromide in the presence of base. The alkene was hydrolyzed overnight with HC1 in THF/H2O, then oxidized with PCC to give an aldehyde. Sodium azide was then added alpha to the carbonyl with KHMDS and TIBSN3 at lowered temperature. The intermediated was then treated with p- OMePhMgBr in THF and toluene to give an intermediate alcohol, which was then reduced using Et3SiH in the presence of BF3-Et2O at reduced temperature. The resulting azide was then reduced with Lindlar’s catalyst under a hydrogen atmosphere to give the corresponding amine, which was acylated with trifluoroacetic acid in pyridine. The benzyl groups were then removed with Pd(OH2) on carbon in MeOH at reflux, and the resulting tri-ol protected as an acetal with dimethoxypropane and camphorsulfonic acid at elevated temperature. The remainder of the synthesis was carried out as described for previous molecules.
FIGURE 45 shows the synthesis of compound MIF-31-3. Galactosamine hydrochloride was fully protected with acetic anhydride, then treated with allyl alcohol in the presence of BF3 ehtrate to give the allyl intermediate. Treatment with pivaloyl chloride in pyridine gave a di-Piv protected intermediate, which was treated with triflic anhydride and subsequently subjected to hydrolysis in water at elevated temperature. The pivaloyl groups were removed by treatment with NaOMe in MeOH to give the allyl triol intermediate. Subsequent steps were performed as described for previous molecules.
FIGURE 46 shows the synthesis of compound MIF-15-3, which was synthesized using procedures analogous to compounds described above.
FIGURE 47 shows the synthesis of compound MIF-19-3. The molecule is synthesized through a late stage triazole-forming click reaction between the triazide and propiolic acid in methanol in the presence of THPTA, copper sulfate, water, and sodium ascorbate. All other reactions are performed as described above.
FIGURE 48 shows the synthesis of compound MIF- 16-3, which was synthesized using procedures analogous to compounds described above.
FIGURE 49 shows the synthesis of compound MIF-20-3, which was synthesized using procedures analogous to compounds described above.
FIGURE 50 shows the synthesis of compound MIF-14-3, which was synthesized using procedures analogous to compounds described above.
FIGURE 51 shows the synthesis of compound MIF-21-3, which was synthesized using procedures analogous to compounds described above.
FIGURES 52-66 show the synthesis of a number of MIF-binding compounds with various ASGPRBM moieties. These are synthesized through methods analogous to those laid out above.
Examples
The instant specification further describes in detail by reference to the following experimental examples. These examples are provided for purposes of illustration only, and are not intended to be limiting unless so specified. Thus, the instant specification should in no way be construed as being limited to the following examples, but rather, should be construed to encompass any and all variations which become evident as a result of the teaching provided herein.
Proper protein section and turnover is a necessary process for maintaining homeostasis. Newly synthesized proteins targeted for secretion are first trafficked to the endoplasmic reticulum, where they are post-translationally modified with N-linked glycan chains terminating in sialic acids(l). As proteins age, terminal sialic acid residues are removed by circulating endogenous glycosydases(2). This natural protein aging process unmasks galactose and N-acetylgalactose (GalNAc) residues, which bind the asialoglycoprotein receptor (ASGPR) on the surface of hepatocytes(3-5).
The ASGPR is a C-type lectin that removes aged circulating proteins with exposed GalNAc residues from circulation by trafficking them to lysosomes. Multiple galactose or GalNAc residues displayed on the protein surface are necessary for high-affinity binding to - and subsequent endocytosis by - ASGPR(6, 7). Once these proteins are endocytosed, they are released from the ASGPR through depletion of calcium from the endosome and changes in binding site amino acid protonation changes due to a decrease in pH (12); the ASGPR is recycled back to the hepatocyte surface(13). Endocytosed proteins are trafficked to late endosomes, which are fused with lysosomes. Lysosomal proteases then degrade endocytosed proteins, permanently removing them from circulation(14). Non-glycosylated proteins are not known to be natural target for the ASGPR. One such protein is macrophage inhibitory factor (MIF), a 12.5 kDa protein with possible catalytic activity (15). Genetic depletion or antibody neutralization of MIF has been shown to have positive results in models of sepsis(16), multiple schlorosis(17), rheumatoid arthritis(18), and burn recovery(21). A bifunctional molecule for degrading cicrulating MIF that takes advantage of ASGPR as an entryway for proteins into the endosomal-lysosomal degradation pathway was proposed. The bicyclic ASGPR-binding molecules in MIF-AcF2 and MIF- AcF3 have been reported previously as high affinity binders for the ASGPR (22, 23).
Example 1: Biological Data
In Example 1, exemplary compounds according to some embodiments were tested to determine their biological activity. Active compounds are shown in FIGURES 1, 7 and 13 hereof. The results of the biological experiments are described herein below.
FIGURE 1 shows exemplary compounds according to the present disclosure. Note that the figure discloses compound 3w (negative control for MIF inhibition), MIF-NVS- PEGnGN3, MIFGN3, MIF-PEGnGN3, MIF-AcF3-l, MIF-AcF3-2 and MIF-AcF3-3. Note that n in the PEG linker sometimes ranges from 1-12, such as 1 to 10, 2 to 8, 2 to 6, 2 to 5 or 1, 2, 3 or 4.
In an experiment the results of which are shown in FIGURE 2, A. fluorescence polarization data of MIF-FITC binding to human MIF indicates that the MIF-binding moiety of the present disclosure binds MIF. B. Bifunctional molecules WJ-PEG4-GN3, WJ-PEG2- GN3, and NVS-PEG3-GN3 bound competitively with MIF-FITC, indicating that the bifunctional molecules maintain the ability to bind human MIF.
In an experiment the results of which are presented in FIGURE 3, bifunctional molecules were able to deplete human MIF from the supernatant of culture HepG2 cells. Briefly, human MIF (100 nM) was added to cell culture media in the presence of negative control MIF inhibitor 3w as well as bifunctional molecules MIF-NVS-PEGn-GN3, MIF- GN3, MIF-PEGn-Gn3, MIF-AcF3-l, MIF-AcF3-2, and MIF-AcF3-3. All molecules utilized a known MIF-binding ligand. Experiments were performed in 96 well plates (approximate surface area .3 cm2). HepG2 cells were grown to 90% confluency in RPMI media, then washed with PBS (2x) and treated with serum-free media (optimem + .1% BSA, + Pen/Strep) containing 100 nM huMIF (Cayman Chemical) and compounds (when applicable). Compounds were diluted from ImM stock solutions in DMSO. After 24 hours, a sample of the supernatant (2 uL) was collected, diluted 1 TOO, and analyzed for MIF content by sandwich ELISA (and incubated for 24 hours in the presence or absence of compound). Remaining MIF levels were determined by sandwich ELISA (biolegend monoclonal anti- MIF and biotinylated anti-MIF antibodies). Data represents the average of at least 3 biological replicates, and error bars represent a standard deviation. After 24 hours, up to 95.3% of the MIF had been depleted from cell culture media (in the case of MIF-AcF3-3).
FIGURE 4 shows the results of an experiment to determine whether or not MIF internalized by HepG2 cells is trafficked to lysosomes. In this experiment, cells were incubated with rhuMIF (Cayman) at a concentration of 100 nM with 200 nM MIF-GN3. After 12 hours, cells were fixed with formaldehyde, permeabilized, and probed with anti- Lamp2 antibody (mouse monoclonal, Abeam), polyclonal rabbit anti-MIF antibody (Thermo) and with Alexa-488 labeled anti-mouse antibody and Alexa 568-labeled anti-rabbit antibody, evidencing internalization in lysosomes.
FIGURE 5 shows that MIF-GN3 mediates the depletion of injected human MIF from mice. Human MIF has a half-life of approximately 40 minutes in mice. In this experiment, human recombinant MIF (Cayman chemical) was co-injected into mice with an anti-DNP IgG, which was used as an injection positive control. In particular, nude mice were injected with 5 pg recombinant human MIF and 200pg anti-DNP IgG as an injection control (FIGURE 4). MIF-GN3 was then injected at the concentration shown and blood drawn every twenty minutes over the course of two hours. Serum was diluted 1 : 100 and analyzed for MIF content by sandwich ELISA (biolegend monoclonal anti-MIF and biotinylated anti-MIF antibodies). The levels of the injected IgG were not significantly different between testing groups. In the mice treated with MIF-GN3, a moderate increase in huMIF levels up to 20 ng/ml was seen, while in mice injected with PBS negative control, serum levels of up to 150 ng/ml were observed, evidencing a substantial decrease in huMIF levels as a consequence of the administration of MIF-GN3.
FIGURE 6 shows that MIF-GN3 is able to delay tumor growth in a mouse model of prostate cancer. In this experiment, nude mice were engrafted with PC3 human prostate cancer cells. Treatment was then initiated immediately with either a non-bifunctional MIF inhibitor (3w), an anti-MIF antibody, or MIF-GN3. MIF-GN3 showed a slowing of tumor growth over the course of the experiment, comparable to the MIF-neutralizing antibody. 3w did not inhibit tumor growth, validating the necessity of degrading MIF for therapeutic efficacy.
FIGURE 7 shows molecules DNP-GN3 and DNP-AcF3-3, which are bifunctional molecules that bind to anti-DNP IgG and ASGPR. These compounds were used in several of the experiments as described below.
FIGURE 8 shows that DNP-GN3 and DNP-AcF3-3 mediate the formation of a ternary complex between HepG2 cells and anti-DNP, thus validating the bifunctional character of the molecules. In this experiment, ASGPR-expressing HepG2 cells were incubated with bifunctional molecules and alexa-488 labeled anti-DNP (Thermo). The readout is mean fluorescence intensity of the cell population. Fluorescence was measured using a flow cytometer.
In a further experiment, the results presented in FIGURE 9 show that DNP-GN3 and DNP-AcF3-3 mediate the uptake of alexa 488-labeled anti-DNP by HepG2 cells. The assay carried out in this experiment was as is described above for MIF uptake. Readout is percentage of Alexa 488-positive cells after 6 hours. Fluorescence was measured using a flow cytometer.
FIGURE 10 shows that DNP-GN3 and DNP-AcF3-3 mediate the localization of alexa 568 labeled anti-DNP to late endosomes and lysosomes. This experiment was carried out as described above for the MIF colocalization studies.
The experimental results presented in FIGURE 11 show that DNP-AcF3-3 mediates the degradation of alexa 488-labeled anti-DNP in HepG2 cells. In this experiment, cells were incubated with 1 uM alexa 488-labeled anti-DNP (Thermo) and 200 nM DNP-AcF3-3. Cells were lysed (RIP A in PBS, containing protease inhibitors) at the given time and assayed by SDS-PAGE gel. Readout is fluorescence of protein fragments.
The results presented in FIGURE 12 evidence that DNP-GN3 mediates the depletion of anti-DNP from mouse serum. Mice were injected with anti-DNP on day 0, then treated with the given compounds each day for 6 days. Serum IgG levels were measured by ELISA. DNP-(OH)3 is used as a non-bifunctional control molecule.
FIGURE 13 shows the structures of IgG-degrading molecules IBA-GN3, Triazine- GN3, FcIII-GN3, and FcIII-4c-GN3.
FIGURE 14 shows that FcIII-GN3 mediates the uptake of human IgG into HepG2 cells. This experiment was performed as described above.
FIGURE 15 shows that FcIII-GN3 mediates the localization of IgG to late endosomes in HepG2 cells. Experiment performed as described above.
Example 2: Experimental Chemistry
In Example 2, synthesis and characterization of exemplary compounds according to some embodiments will be described. Example 2-1. MIF binding molecule (Figure 13) Example 2-1-1: MIF-1
Figure imgf000267_0001
2-chloroquinolin-6-ol (1.00 g, 5.57 mmol) and K2CO3 (1.53 g, 11.1 mmol, 2.0 eq) were dissolved in DMF (20 mL). Ethyl bromobutyrate (1.63 g, 1.2mL, 8.35 mmol, 1.5eq) was then added and the mixture stirred at 80º for 12 hours. The reaction was diluted into ethyl acetate and washed with water (2x) and brine (3x). The organic layer was dried over sodium sulfate and evaporated to give compound 30, which was used in the next step without further purification. 1H NMR (400 MHz, Chloroform-d) δ 7.98 (d, J = 8.6 Hz, 1H), 7.92 (d, J = 9.2 Hz, 1H), 7.40 – 7.32 (m, 2H), 7.07 (d, J = 2.7 Hz, 1H), 4.20 – 4.09 (m, 5H), 2.56 (t, J = 7.2 Hz, 2H), 2.19 (t, J = 6.7 Hz, 2H), 1.26 (t, J = 7.1 Hz, 4H). 13C NMR (101 MHz, cdcl3) δ 173.24, 157.46, 148.18, 143.87, 137.83, 130.05, 128.06, 123.40, 122.67, 106.20, 77.48, 77.16, 76.84, 67.30, 60.69, 30.87, 24.63, 14.39. HRMS: [M+H]+ Expected 294.090, found 294.11 Example 2-1-2: MIF-2
Figure imgf000267_0002
Compound 30 (1.52 g, 5.17 mmol) was dissolved in THF (20 mL) and triethylamine (2.88 mL, 20.7 mmol, 4 eq). Copper (I) iodide (49.0 mg, .258 mmol, .05 eq), Pd(PPh3)2Cl2 (181 mg, .258 mmol, .05 eq), and TMS-acetylene (1.07 mL, 762 mg, 7.75 mmol, 1.5 eq) were then added and the reaction was stirred under pressure at 65º for 16 hours. The reaction mixture was filtered through celite, washed ethyl acetate, and evaporated. The residue was purified on silica (50% ethyl acetate in hexanes) to give compound 31. 1H NMR (500 MHz, Chloroform-d) δ 8.00 (d, J = 8.4 Hz, 2H), 7.50 (d, J = 8.5 Hz, 1H), 7.43 – 7.33 (m, 1H), 7.04 (d, J = 2.2 Hz, 1H), 4.15 (dt, J = 12.7, 6.5 Hz, 4H), 2.56 (t, J = 7.2 Hz, 2H), 2.24 – 2.13 (m, 2H), 1.26 (t, J = 7.1 Hz, 3H), 0.30 (s, 9H). 13C NMR (151 MHz, cdcl3) δ 173.07, 124.73, 105.65, 67.11, 60.51, 30.67, 24.42, 14.21, -0.27. HRMS: [M+H]+ Expected 356.168, Found 356.505 Example 2-1-3: MIF-3
Figure imgf000268_0001
Procedure Compound 31 (1.57 g, 4.42 mmol) was dissolved in DCM (45 mL) and TBAF (5.3 mL, 1M in THF, 5.30 mmol, 1.2 eq) was added dropwise. After 1 minute of stirring 10% citric acid (50 mL) was added and the reaction stirred for 30 minutes. The organic phase was washed with water (1x), dried, and evaporated to give compound 32, which was used in the next step without further purification. 1H NMR (600 MHz, Chloroform-d) δ 8.11 – 8.00 (m, 1H), 7.51 (d, J = 8.4 Hz, 1H), 7.38 (dd, J = 9.3, 2.3 Hz, 1H), 7.05 (d, J = 2.4 Hz, 1H), 4.15 (p, J = 6.6, 6.0 Hz, 3H), 3.43 – 3.36 (m, 1H), 2.56 (t, J = 7.2 Hz, 1H), 2.22 – 2.14 (m, 1H), 1.73 – 1.64 (m, 1H), 1.47 (q, J = 7.4 Hz, 1H), 1.26 (t, J = 7.1 Hz, 2H), 1.02 (t, J = 7.3 Hz, 1H). CNMR: 13C NMR (151 MHz, cdcl3) δ 173.06, 137.60, 129.91, 128.64, 124.50, 123.18, 122.48, 105.99, 105.58, 77.20, 76.99, 76.77, 67.12, 60.51, 59.14, 30.67, 24.42, 24.22, 19.80, 14.21, 13.69. HRMS: [M+H]+ Expected 284.129, Found 284.327
Figure imgf000268_0002
2-fluoro-4-iodophenol (126 mg, .529 mmol) and sodium azide (38 mg, .528 mmol, 1.0 eq) were dissolved in DMSO (2.5 mL) and stirred for two hours at 70º. Compound 32 (150 mg, .529 mmol, 1 eq), trans-N,N’-dimethylcyclohexane-1,2-diamine (11 mg, .079 mmol, .15 eq), sodium ascorbate (10 mg, .053 mmol, .1 eq), copper (I) iodide (15 mg, .079 mmol, .15 eq), and H2O (2.5 mL) were then added, and the mixture stirred at 70º overnight. The reaction was diluted with ethyl acetate and washed with H2O (1x) and brine (1x). The organic layer was dried over sodium sulfate, evaporated, and purified on silica (DCM/EtOAc) to give compound 33. 1H NMR (600 MHz, DMSO-d6) δ 10.46 (s, 1H), 9.32 (s, 1H), 8.39 (d, J = 8.6 Hz, 1H), 8.23 (d, J = 8.5 Hz, 1H), 7.95 (dd, J = 11.6, 2.6 Hz, 2H), 7.77 – 7.71 (m, 1H), 7.43 (dd, J = 4.8, 2.0 Hz, 2H), 7.16 (t, J = 9.0 Hz, 1H), 4.17 (d, J = 6.3 Hz, 2H), 4.13 – 4.07 (m, 3H), 3.17 (d, J = 5.2 Hz, 3H), 2.53 (d, J = 7.3 Hz, 3H), 2.07 (t, J = 6.8 Hz, 2H), 1.19 (d, J = 7.1 Hz, 3H), 0.94 (d, J = 7.3 Hz, 1H). 13C NMR (151 MHz, dmso) δ 172.96, 156.95, 151.95, 150.34, 148.62, 145.98, 143.82, 136.47, 130.40, 129.01, 123.19, 121.83, 119.06, 118.62, 117.31, 109.87, 109.72, 107.12, 67.43, 60.35, 49.03, 40.48, 40.36, 40.22, 40.09, 39.95, 39.81, 39.67, 39.53, 30.61, 24.58, 23.48, 14.56, 13.93. HRMS: Expected 437.163, Found 437.164 Example 2-1-5: MIF-5
Figure imgf000269_0001
Compound 33 (90 mg, .206 mmol) was dissolved in dioxane (6 mL) and 2M NaOH (3 mL). The reaction was stirred for 2.5 hours at room temperature, at which time the reaction was diluted with water and the pH adjusted to 3-4 with 1M HCl. The mixture was cooled to 4º and filtered to give compound 34, which was used without further purification. Example 2-2. GaINAc spacer (Figure 14)
Figure imgf000269_0002
Triethylene glycol (17.5 mL, 19.7 g, 131.13 mmol, 5 eq) was dissolved in DCM (150 mL) and trimethylamine (5.48 mL, 3.98 g, 1.5 eq) and cooled to 0º. TsCl (5.00 g, 26.23 mmol, 1 eq) was then added and the reaction mixture stirred at room temperature for 18 hours. The reaction was diluted into DCM and washed with water (3x) and brine (1x). The organic layer was dried over sodium sulfate and concentrated in vacuo. The crude product was purified on silica (0-5% MeOH in DCM) to give compound 64 (6.89 g, 22.6 mmol) in 85% yield. 1H NMR (400 MHz, Chloroform-d) δ 7.80 (d, J = 8.3 Hz, 2H), 7.38 – 7.30 (m, 2H), 4.23 – 4.14 (m, 2H), 3.71 (td, J = 5.3, 4.3 Hz, 4H), 3.66 – 3.55 (m, 6H), 2.45 (s, 3H). 13C NMR (101 MHz, cdcl3) δ 144.98, 133.09, 129.95, 128.09, 72.58, 70.91, 70.44, 69.28, 68.84, 61.88, 21.76.
Figure imgf000270_0001
Compound 64 (2.00 g, 6.57 mmol) and sodium azide (.470 g, 7.23 mmol, 1.1 eq) were dissolved in DMF (40 mL) and stirred overnight at 60º.25 mL of DMF was then removed by rotary evaporation, and the resulting mixture diluted into water and extracted with ethyl acetate (2x). The organic layers were washed with brine (3x), dried over sodium sulfate, and evaporated. The crude product was purified on silica (0-5% MeOH in DCM) to give compound 65 (932 mg, 5.32 mmol) in 81% yield. Example 2-3. GaINAc ASGPR ligand (Figure 15)
Figure imgf000270_0002
[0001] Galactosamine pentaacetate (100 mg, .257 mmol) was dissolved in dichloroethane (1 mL) and stirred at room temperature before the addition of TMSOTf (70 uL, 86.0 mg, .387 mmol, 1.5 eq). The reaction was stirred at 50º for 90 minutes, then allowed to cool to room temperature and stirred for a further 12 hours. The reaction was poured into ice cold saturated sodium bicarbonate and extracted into DCM. The organic layer was washed with water (2x), dried over sodium sulfate, and evaporated to give compound 66 (.236 mmol, 77.7 mmol, 92%) as a dark gum, which was used without further purification. 1H NMR (400 MHz, Chloroform-d) δ 5.98 (d, J = 6.8 Hz, 1H), 5.45 (t, J = 3.0 Hz, 1H), 4.90 (dd, J = 7.4, 3.3 Hz, 1H), 4.29 – 4.20 (m, 1H), 4.17 (d, J = 6.9 Hz, 1H), 4.10 (dd, J = 11.1, 5.7 Hz, 1H), 3.99 (td, J = 7.1, 1.4 Hz, 1H), 2.11 (s, 3H), 2.05 (m, J = 7.6 Hz, 6H). 13C NMR (101 MHz, cdcl3) δ 170.46, 170.13, 169.78, 166.35, 121.82, 118.64, 101.41, 71.76, 69.44, 65.25, 63.53, 61.56, 46.82, 20.77, 20.68, 20.54, 14.41, 8.64, -0.06.
Figure imgf000271_0001
Compound 66 (200 mg, .607 mmol) and compound 65 (160 mg, .913 mmol, 1.5 eq) were dissolved in 1,2-dichloroethane (5 mL).4 Å molecular sieves were then added, and the reaction stirred for 30 minutes. TMSOTf (55 uL, 67.5 mg, .304 mmol, .5 eq) was then added to the mixture, and the reaction stirred overnight. The mixture was diluted into DCM, washed with 1M sodium bicarbonate (1x) and water (1x), then dried over magnesium concentrate and concentrated. The curde oil was purified on silica gel (50-100% EtoAc in DCM) to give compound 67 (245 mg, .486 mmol) in 80.1% yield. 1H NMR (400 MHz, Chloroform-d) δ 6.13 (d, J = 9.3 Hz, 1H), 5.31 (dd, J = 3.4, 1.1 Hz, 1H), 5.05 (dd, J = 11.2, 3.4 Hz, 1H), 4.77 (d, J = 8.6 Hz, 1H), 4.28 – 4.04 (m, 3H), 3.93 – 3.79 (m, 3H), 3.78 – 3.58 (m, 8H), 3.43 (dt, J = 28.2, 4.9 Hz, 4H), 2.15 (s, 3H), 2.04 (s, 3H) 1.98 (s, 3H), 1.97 (s, 3H) 13C NMR (101 MHz, cdcl3) δ 170.24, 170.09, 169.99, 101.84, 72.06, 71.27, 70.50, 70.29, 70.27, 70.20, 70.00, 69.98, 69.67, 69.37, 68.18, 66.30, 61.38, 61.17, 50.32, 50.26, 50.11, 22.80, 20.37, 20.31 HRMS: [M+Na]+ 527.207 found 527.203
Figure imgf000271_0002
Compound 67 (1.80 g, 3.57 mmol) was dissolved in THF (35 mL). Triphenylphosphine (1.40 g, 5.35 mmol, 1.5 eq) and water (257 uL, 14.28 mmol, 4 eq) were then added and the reaction stirred at room temperature under nitrogen for 36 hours. The solvent was removed and the crude product used in the next step without further purification. Example 2-4. Tris Valent Glycine (Figure 16)
Figure imgf000271_0003
Tris base (5.00 g, 41.3 mmol) was dissolved in dichloromethane (80 mL) and trimethylamine (20 mL). Di-tert-butyl dicarbonate (10.81 g, 49.6 mmol, 1.2 eq) was then added, and the reaction stirred for 4 hours. The mixture was evaporated and the residue portioned between ethyl acetate and water. The organic fraction was washed with water (lx), IM HC1 (2x), saturated sodium bicarbonate (lx), and brine (lx) before drying over sodium sulfate and evaporation to give compound 23 (9.04g, 40.9 mmol) in 99% yield, which was used without purification in further steps.
Figure imgf000272_0001
Compound 27 (9.04 g, 40.9 mmol) was dissolved in a mixture of dioxane (17 mL) and aqueous KOH (4.98 g, 88.7 mmol, 2.46 mL). Acrylonitrilte (8.84 mL, 7.16 g, 135.0 mmol, 3.3 eq) was then added dropwise over a period of 2.5 hours, and the reaction stirred under nitrogen for 24 hours. The reaction was neutralized with the addition of 2M HC1 (30 mL) and portioned between DCM and water. The organic layer was washed with water (2x) and brine (lx), dried over sodium sulfate, and evaporated. The crude mixture was purified on silica (0- 80% EtOAc in hexanes) to give compound 20 (7.87g, 20.7 mmol) in 59% yield.
HNMR: ’H NMR (400 MHz, Chloroforms/) 8 4.84 (s, 1H), 3.73 (s, 6H), 3.65 (t, J= 6.1 Hz, 6H), 2.57 (t, J= 6.1 Hz, 6H), 1.35 (s, 9H).
Figure imgf000272_0002
Compound 28 (7.87 g, 20.7 mmol) was dissolved in MeOH (40 mL) and concentrated sulfuric acid (10 mL) was added. The reaction was stirred at reflux under nitrogen for 24 hours, then neutralized with sodium bicarbonate. Methanol was evaporated, and the residue pardoned between water and ethyl acetate. The ethyl acetate layer was washed with sodium bicarbonate (lx) and brine (lx), then dried over sodium sulfate. The crude residue was purified on silica (10% MeOH in DCM) to give compound 29 (5.50 g, 14.5 mmol) in 70% yield. HNMR: 1H NMR (400 MHz, Chloroform-d) δ 3.79 – 3.64 (m, 15H), 3.32 (s, 6H), 2.56 (t, J = 6.3 Hz, 6H). CNMR: 13C NMR (101 MHz, cdcl3) δ 172.03, 72.52, 66.77, 56.10, 51.62, 34.80.
Figure imgf000273_0001
Compound 70 (723 mg, 1.90 mmol) was dissolved in MeCN (25 mL). HOBT (291 mg, 1.90 mmol, 1 eq), Cbz-glycine (397 mg, 1.90 mmol, 1 eq), and DCC (392 mg, 1.90 mmol, 1 eq) were then added, and the reaction stirred overnight. MeCN was then evaporated, and the residue adsorbed onto silica and purified using a gradient of 0-75% EtOAc in hexanes. Compound 71 (866 mg, 1.52 mmol) was recovered in 80% yield. 1H NMR (600 MHz, Chloroform-d) δ 7.33 (dd, J = 24.9, 4.4 Hz, 5H), 6.33 (s, 1H), 5.55 (s, 1H), 5.12 (s, 2H), 3.86 (d, J = 5.1 Hz, 2H), 3.68 (t, J = 5.5 Hz, 21H), 3.49 (s, 1H), 2.53 (t, J = 6.1 Hz, 6H). 13C NMR (151 MHz, cdcl3) δ 172.14, 168.67, 156.32, 136.41, 128.45, 128.05, 127.98, 69.04, 66.85, 66.71, 59.83, 51.69, 44.55, 34.64. Expected: [M+H]+ 571.250, found 571.243
Figure imgf000273_0002
Compound 71 (100 mg, .175 mmol) was dissolved in dioxane (2 mL) and 2M NaOH (2mL). The reaction was stirred for 3 hours, then acidified and extracted twice into ethyl acetate. The organic fraction was washed with 1M HCl, then dried over sodium sulfate and evaporated to give compound 72, which was used in further steps without purification. Example 2-5. GaINAc trivalent (Figure 17)
Figure imgf000274_0001
Compound 72 (372 mg, .704 mmol, 1 eq) was dissolved in DMF (40 mL) and DIPEA (981 uL, 728 mg, 5.632 mmol, 8 eq). HBTU (1.01 g, 2.67 mmol, 3.8 eq) was then added, and the reaction stirred for 10 minutes at room temperature before the addition of compound 68 (1.28 g, 2.67 mmol, 3.8 eq). The reaction was stirred for two hours, then diluted into DCM and washed with H3PO4 (1M, 1x), NaHCO3 (1M, 1x), and brine (1x). The organic layer was dried over sodium sulfate and evaporated onto silica. The residue was purified (0-20% MeOH in DCM) to give compound 73 (831 mg, .436 mmol) in 62% yield. 1H NMR (500 MHz, DMSO-d6) δ 7.91 (t, J = 5.7 Hz, 3H), 7.80 (d, J = 9.2 Hz, 3H), 7.39 - 7.28 (m, 6H), 7.12 (s, 1H), 5.21 (d, J = 3.4 Hz, 3H), 5.02 (s, 2H), 4.97 (dd, J = 11.2, 3.4 Hz, 3H), 4.55 (d, J = 8.5 Hz, 3H), 4.10 – 3.99 (m, 9H), 3.92 – 3.84 (m, 3H), 3.81 - 3.75 (m, 3H), 3.62 - 3.46 (m, 35H), 3.39 (t, J = 6.1 Hz, 6H), 3.23 – 3.16 (m, 6H), 2.30 (t, J = 6.4 Hz, 6H), 2.10 (s, 9H), 1.99 (s, 9H), 1.89 (s, 9H), 1.77 (s, 9H). 13C NMR (126 MHz, dmso) δ 170.40, 170.16, 170.09, 169.79, 169.48, 169.04, 156.60, 137.23, 128.50, 127.94, 127.84, 101.12, 72.50, 70.65, 70.07, 69.93, 69.86, 69.77, 69.59, 69.29, 68.50, 67.51, 66.89, 65.59, 61.63, 60.38, 59.84, 49.50, 43.77, 38.68, 36.01, 22.95, 20.68, 20.62, 20.60.
Figure imgf000275_0001
Compound 73 (710 mg, .372 mmol) was dissolved in dry methanol (90 mL) and cooled to 0° under nitrogen. Pd/C (71.0 mg, 10% w/w) was then added, and the reaction stirred under hydrogen (1 atm) at 0° for 16 hours. Upon completion, the reaction was filtered through celite and methanol evaporated to give compound 74 (657 mg, .370 mmol) in 99.5% yield, which was used without further purification.
Compound 74 (441 mg, .248 mmol) was dissolved in methanol (15 mL) and cooled to 0°. Sodium methoxide solution (400 uL, 5.4M in MeOH) was then added, and the reaction stirred for 30 minutes. Dowex 50WX8 was then added until the solution was weakly acidic. The resin was filtered off and washed thoroughly with methanol. The combined methanol fractions were evaporated under reduced pressure to give compound 75 (274 mg, .196 mmol) in 79% yield. Compound 75 was used in further steps without purification.
Example 2-6. MIF-GN3 (Figure 18)
Figure imgf000276_0001
Compound 34 (23.5 mg, .0575 mmol, 1.1 eq) and HATU (20.0 mg, .0522 mmol, 1 eq) were dissolved in dry DMF (5 mL) and DIPEA (23.3 uL, 16.9 mg, .131 mmol, 2.5 eq) and stirred for 10 minutes at room temperature. Compound 75 (73.0 mg, .0522 mmol) was then added, and the reaction stirred for 30 minutes. The mixture was loaded directly onto HPLC and purified (20-30% MeCN in water, 3% TFA) to give compound 76 (12 mg, .0067 mmol) in 12.8% yield.
Expected [M+H]+ 1787.801, found 1787.823
Example 2-7. Bicyclic ASGPR Spacer (Figure 19)
Figure imgf000276_0002
Tetraethylene glycol (50.0 g, 258 mmol) was dissolved in THF (1 mL), cooled to 0°, and stirred. NaOH (1.65 g, 41.3 mmol, 1.6eq) in water (1 mL) was then added, followed by the dropwise addition of p-toluenesulfonyl chloride (4.92 g, 25.8 mmol, 1 eq) in THF (3 mL). The reaction mixture was stirred at 0° for 4 hours, then diluted into DCM. The organic layer was washed with ice-cold water (2x), brine (lx), and dried over sodium sulfate to give compound 16 (8.84 g, 25.4 mmol, 99% yield), which was used in further steps without purification.
Figure imgf000276_0003
Compound 16 (8.84 g, 25.4 mmol) was dissolved in 100% ethanol (200 mL) and sodium azide (4.128 g, 63.5 mmol, 2.5 eq) was added. The reaction was heated to reflux for 16 hours, then cooled to room temperature before the addition of water (150 mL). Ethanol was then evaporated under reduced pressure and the product extracted into ethyl acetate (2x). The organic layer was washed with water (1x) and brine (1x), dried over sodium sulfate, and evaporated to give compound 17 (4.82 g, 22.1 mmol) as a yellow oil in 87% yield. HNMR: 1H NMR (400 MHz, Chloroform-d) δ 3.69 – 3.64 (m, 2H), 3.61 (m, J = 4.2 Hz, 10H), 3.57 – 3.51 (m, 2H), 3.33 (td, J = 5.0, 2.3 Hz, 2H), 2.81 (s, 1H). CNMR: 13C NMR (101 MHz, cdcl3) δ 72.47, 70.65, 70.63, 70.59, 70.52, 70.28, 69.99, 61.60, 50.59.
Figure imgf000277_0001
Compound 17 (5.00 g, 22.8 mmol) was dissolved in pyridine (50 mL). Methanesulfonyl chloride (3.14 g, 27.4 mmol, 1.2 eq) was then added and the reaction stirred for six hours under nitrogen. The mixture was then diluted into ethyl acetate, washed with water (3x), .5M HCl (2x), saturated sodium bicarbonate (1x), and brine (1x), dried over sodium sulfate, and evaporated to give compound 18 (5.71 g, 19.2 mmol) in 84% yield. HNMR: 1H NMR (400 MHz, Chloroform-d) δ 4.41 – 4.33 (m, 2H), 3.81 – 3.72 (m, 2H), 3.70 – 3.59 (m, 10H), 3.38 (t, J = 5.0 Hz, 2H), 3.06 (s, 3H). CNMR: 13C NMR (101 MHz, cdcl3) δ 70.79, 70.75, 70.71, 70.15, 69.39, 69.11, 50.77, 37.78. HRMS: Expected 298.107, found 298.105 Example 2-8. Bicyclic ASGPR Precursor (Figure 20)
Figure imgf000277_0002
Pentaacetyl galactose (25.0 g, 64.0 mmol) was dissolved in 33% HBr in HOAc (30 mL) and stirred under nitrogen for 2 hours. The reaction was diluted into EtOAc (500 mL) and washed with water (3x), saturated sodium bicarbonate (1x), and brine (1x). The organic layer was dried over sodium sulfate and evaporated to give compound 1 as a pale yellow oil in quantitative yield. The compound was used without further purification. Compound 1 (26.34 g, 64.06 mmol) was dissolved in acetic acid (510 mL) and zinc (67.01 g, 1024 mmol) added. The mixture was stirred vigorously. A solution of CuSO4 (2.96 g, 18.6 mmol) in aqueous NaH2PO4 (128 mL, .1M, 1.53 g) was then added, and the reaction stirred for 1 hour. The reaction mixture was filtered over celite and the resulting water/AcOH mixture evaporated to give a white solid. The white solid was dissolved in EtOAc (2x, 300 mL each), water (300 mL), and EtOAc (1x, 300 mL). The layers were separated and the organic layer further washed with water (2x), saturated sodium bicarbonate (2x), and brine (1x). The organic solution was dried over sodium sulfate, evaporated, and purified on silica (15-25% EtOAc in Hexanes) to give compound 2 in 84% yield (14.64 g, 53.76 mmol). 1H NMR (400 MHz, Chloroform-d) δ 6.46 (dd, J = 6.3, 1.8 Hz, 1H), 5.54 (qd, J = 2.8, 1.2 Hz, 1H), 5.41 (dt, J = 4.7, 1.7 Hz, 1H), 4.71 (ddd, J = 6.3, 2.7, 1.5 Hz, 1H), 4.33 (ddt, J = 7.0, 5.6, 1.4 Hz, 1H), 4.29 – 4.15 (m, 2H), 2.11 (s, 3 H), 2.06 (s, 3H), 2.00 (s, 3H). 13C NMR (101 MHz, Chloroform-d) δ 170.33, , 170.06, 169.97, 145.30, 98.80, 72.69, 63.82, 63.59, 61.85, 20.66, 20.64, 20.58. HRMS: [M+Na]+ Expected 295.079, found 295.078
Figure imgf000278_0001
Procedure: Compound 2 (12.0 g, 44 mmol) was dissolved in acetonitrile (250 mL) and cooled to -10º. In a separate nitrogen-flushed flask at -10º, NaN3 (4.3 g, 66 mmol) and ceric ammonium nitrate (87.0 g, 158 mmol) were mixed and stirred vigorously. The solution of compound 2 in acetonitrile was added dropwise via cannula, and the mixture allowed to slowly reach room temperature. The reaction mixture was allowed to stir for a total of 12 hours before dilution with ethyl acetate (500 mL) and washing with water (3x) and brine (1x). The organic layer was dried over sodium sulfate, evaporated, and purified on silica (20-50% EtOAc in Hexanes) to give compound 3 in 79% yield (13.1g, 34.9 mmol). 1H NMR (400 MHz, Chloroform-d) δ 6.31 (d, J = 4.1 Hz, 1H), 5.60 (d, J = 8.8 Hz, 1H), 5.43 (dd, J = 3.4, 1.3 Hz, 1H), 5.32 (t, J = 3.3 Hz, 1H), 5.16 (dt, J = 11.6, 3.4 Hz, 1H), 4.96 (dd, J = 10.6, 3.3 Hz, 1H), 4.39 – 4.28 (m, 1H), 4.14 – 4.00 (m, 5H), 3.76 (ddd, J = 13.5, 9.6, 4.7 Hz, 1H), 2.19 – 2.05 (m, 6H), 2.05 – 1.88 (m, 12H). 13C NMR (101 MHz, cdcl3) δ 170.15-169.06, 97.91, 97.79, 96.91, 71.68, 71.45, 69.35, 68.42, 66.98, 66.53, 65.85, 64.75, 61.07, 60.84, 57.38, 55.82, 55.08, 20.31-20.20. HRMS: [M+Na]+ expected 399.076, found 399.073
Figure imgf000279_0001
Sodium methoxide solution was prepared from ice-cold dry methanol (50 mL) and sodium hydride (2.296 g, 95.67 mmol) 3 eq) and added to a solution of compound 3 (12.00 g, 31.89 mmol) in dry methanol (100 mL). After thirty minutes of stirring, the reaction was confirmed neutralized by the addition of acetic acid and directly loaded onto silica gel. The reaction mixture was purified over a gradient of 0-20% MeOH in DCM to give compound 4 (6.64 g, 30.3 mmol) in 95% yield. CNMR: 13C NMR (101 MHz, cd3od) δ 102.60, 100.87, 98.59, 75.65, 74.62, 71.43, 70.41, 68.82, 67.81, 67.69, 67.47, 67.37, 63.73, 62.96, 60.75, 60.43, 59.54, 55.42, 53.69, 45.94. HRMS: [M+Na]+ expected 242.075, found 242.072
Figure imgf000279_0002
Compound 4 (5.00 g, 22.8 mmol) was dissolved in pyridine (100 mL) and stirred under nitrogen. Trimetylsilylchloride (10.43 mL, 8.929 g, 82.18 mmol, 3.6 eq) was added dropwise and the mixture stirred for 6 hours. The reaction was diluted into ethyl acetate and washed with water (2x) and brine (1x). The organic layer was dried over sodium sulfate and evaporated to give the tri-TMS intermediate. Residual pyridine was removed by coevaporating with toluene (3x). The intermediate was taken up into dry MeOH (45 mL) and cooled to 0º before potassium carbonate (40 mg) was added. The reaction was closely monitored over 1.5 hours and quenched with acetic acid (17 uL) once TLC showed complete consumption of starting material. The product was then dry loaded onto silica and purified with a gradient of 0-50% EtOAc in hexane to give compound 5 (6.55 g, 18.0 mmol) in 79% yield. 13C NMR (101 MHz, cdcl3) δ 103.36, 75.29, 73.71, 72.37, 71.41, 70.94, 70.38, 64.04, 62.49, 62.15, 61.05, 60.36, 57.15, 55.17, 34.60, 31.52, 25.21, 22.59, 20.93, 14.11, 14.05, 0.85, 0.57, 0.55, 0.52, 0.22, 0.14, 0.01, -0.07.
HRMS: [M+Na]+ 386.154, found 386.156
Figure imgf000280_0001
Compound 5 (7.00 g, 19.3 mmol) was dissolved in DCM (100 mL) and stirred under nitrogen. Dess-Martin periodane (9.82 g, 23.2 mmol, 1.2 eq) was added and the mixture stirred for 2 hours. The reaction was diluted into DCM and washed with water (2x) and brine (lx). The organic layer was dried over sodium sulfate and evaporated to give the intermediate aldehyde.
Compound 6 was dissolved in molecular sieve-dried EtOH (100 mL). Paraformaldehyde (36.50 g, 384.9 mmol, 20 eq) and 21% sodium ethoxide solution (14.5 mL, 38.5 mmol, 2 eq) were added and the reaction stirred for 8 hours. The solvent was evaporated and the product adsorbed onto silica. The product was purified using a gradient of 0-25% MeOH in DCM to afford compound 7 (2.981 g, 11.97 mmol) in 62% yield.
HRMS: [M+Na]+ expected 272.086 (+Na), found 272.083
Figure imgf000280_0002
L6-7 (500 mg, 2.00 mmol) was dissolved in water (4.5 mL) and sulfuric acid (.5 mL). The reaction was sealed in a microwave vial and heated at 100° for 40 minutes. The reaction was cooled to 0°, then diluted with MeOH (10 mL) and neutralized by the addition of concentrated ammonia solution. Salts were filtered off and washed several times with methanol. The filtrate was adsorbed onto silica and purified on a gradient of 0-15% MeOH in DCM to give compound L6-8 (347 mg, 1.60 mmol) in 80% yield.
13C NMR (101 MHZ, cd3od) 8 102.70, 85.32, 71.03, 69.59, 69.49, 66.07, 61.85
Example 2-9. Bicyclic ASGPR ligand CF3 (Figure 21)
Figure imgf000281_0001
Compound (400 mg, 1.84 mmol) was dissolved in methanol (30 mL) and the reaction flask purged with nitrogen. Lindlar’s catalyst (40.0 mg, 10 wt%) was then added, and the reaction mixture stirred under a 1 atm hydrogen atmosphere (balloon) for 6 hours. The reaction was filtered over celite and evaporated to give compound 10 (351 mg, 1.84 mmol) in quantitative yield, which was used in the next reaction without further purification. Compound (351 mg, 1.84 mmol) was dissolved in pyridine (15 mL) and treated with trifluoroacetic anhydride (1.24 mL, 1.85 g, 8.83 mmol, 4.8 eq). The reaction was stirred for 6 hours, then diluted into ethyl acetate and washed with 1M HCl (1x), saturated sodium bicarbonate (1x), and brine (1x). The organic layer was dried over sodium sulfate and evaporated to give compound 11 (1.01 g, 1.75 mmol) in 95% yield, which was used in further steps without purification. Compound 11 (1.01 g, 1.75 mmol) was dissolved in methanol (25 mL) and dry sodium methoxide (86.2 mg, 1.60 mmol, 4 eq) was added. The reaction was stirred for one hour at room temperature, then neutralized with acetic acid and evaporated onto silica. The crude mixture was purified on silica (0-15% MeOH in DCM) to give compound 12 (482 mg, 1.68 mmol) in 96% yield. 1H NMR (400 MHz, DMSO-d6) δ 9.51 (d, J = 6.6 Hz, 1H), 5.15 (s, 1H), 4.98 – 4.90 (m, 1H), 4.87 (d, J = 5.6 Hz, 1H), 4.75 – 4.70 (m, 1H), 3.86 – 3.80 (m, 2H), 3.80 – 3.70 (m, 2H), 3.67 – 3.54 (m, 3H). 13C NMR (101 MHz, dmso) δ 157.47, 157.11, 117.74, 114.87, 100.28, 84.21, 68.77, 68.28, 66.00, 60.51, 55.95, 40.56, 40.35, 40.15, 39.94, 39.73, 39.52, 39.31. HRMS: [M+H]+ Expected 288.069, found 288.064
Figure imgf000281_0002
Compound 12 (110 mg, .383 mmol) was dissolved in DMF (8 mL) and dimethoxypropane (236 uL, 200 mg, 1.92 mmol, 5 eq) and camphorsulfonic acid (45 mg, .192 mmol, .5 eq) were added. The reaction was stirred at 70º overnight, then DMF evaporated under reduced pressure. The residue was dissolved in ethyl acetate, washed with saturated sodium bicarbonate (1x) and brine (1x), then evaporated onto silica and purified (0- 5% MeOH in DCM) to give compound 13 (99.1 mg, .303 mmol) in 79% yield. HNMR: 1H NMR (400 MHz, Chloroform-d) δ 6.87 (d, J = 9.0 Hz, 1H), 5.35 (d, J = 2.1 Hz, 1H), 4.18 – 4.11 (m, 2H), 4.11 – 4.03 (m, 2H), 3.84 (t, J = 8.2 Hz, 2H), 3.74 (d, J = 7.9 Hz, 1H), 2.66 (d, J = 6.6 Hz, 1H), 1.52 (s, 3H), 1.32 (s, 3H). CNMR: 13C NMR (101 MHz, cdcl3) δ 157.87, 157.50, 157.12, 156.75, 119.93, 117.07, 114.21, 112.04, 111.35, 100.22, 81.55, 75.57, 75.00, 68.44, 60.96, 55.16, 27.67, 26.16. HRMS: [M+H]+ Expected 328.101, found 328.095
Figure imgf000282_0001
Compound 13 (99.1 mg, .303 mmol) was dissolved in DMF (5 mL) and treated with sodium hydride (8.7 mg, .364 mmol, 1.2 eq), then stirred under nitrogen for 15 minutes. Compound 18 (108 mg, .364 mmol, 1.2 eq) was then added, and the reaction stirred for 1 hour. The reaction was neutralized by the dropwise addition of acetic acid. The solvent was removed under reduced pressure and the residue taken up into ethyl acetate and washed with brine (4x), and the organic layer was dried over sodium sulfate and evaporated onto silica. The crude mixture was purified on silica (50-100% EtOAc in hexanes) to give compound 14 (131 mg, .248 mmol) in 82% yield. HNMR: 1H NMR (400 MHz, DMSO-d6) δ 9.75 (d, J = 8.3 Hz, 1H), 5.29 (s, 1H), 4.40 (t, J = 6.7 Hz, 1H), 4.30 (d, J = 5.9 Hz, 1H), 3.88 – 3.66 (m, 5H), 3.64 – 3.48 (m, 14H), 3.39 (t, J = 5.0 Hz, 2H), 1.41 (s, 3H), 1.28 (s, 3H).
Figure imgf000282_0002
Compound 14 (131 mg, .240 mmol) was dissolved in methanol (10 mL) and stirred under a nitrogen atmosphere. Lindlar catalyst (13.1 mg, 10 wt%) was then added, and the reaction stirred for 6 hours under H2 atmosphere (1 atm). The reaction was then filtered over celite and the solvent evaporated to give compound 15 (120 mg, .240 mmol) in quantitative yield, which was used without further purification.
HRMS: [M+H]+ Expected 503.222, found 503.223
Example 2-10. MIE binding divalent (Figure 23)
Figure imgf000283_0001
Serinol (2.00 g, 22.0 mmol) was dissolved in dichloromethane (40 mL) and trimethylamine (10 mL). Di-tert-butyl dicarbonate (5.76 g, 26.4 mmol, 1.2 eq) was then added, and the reaction stirred for 4 hours. The mixture was evaporated and the residue portioned between ethyl acetate and water. The organic fraction was washed with water (lx), IM HC1 (2x), saturated sodium bicarbonate (lx), and brine (lx) before drying over sodium sulfate and evaporation to give compound 23 (3.99 g, 20.9 mmol) in 95% yield, which was used without purification in further steps.
Figure imgf000283_0002
Compound 23 (3.99 g, 20.9 mmol) was dissolved in a mixture of dioxane (12 mL) and aqueous KOH (1.63 g, 29 mmol, 2.4 mL). Acrylonitrilte (3.02 mL, 2.44 g, 46.0 mmol, 2.2 eq) was then added dropwise over a period of 2.5 hours, and the reaction stirred under nitrogen for 24 hours. The reaction was neutralized with the addition of 2M HC1 (16 mL) and portioned between DCM and water. The organic layer was washed with water (2x) and brine (lx), dried over sodium sulfate, and evaporated. The crude mixture was purified on silica (20- 100% EtOAc in hexanes) to give compound 20 (4.96 g, 16.7 mmol) in 80% yield.
HNMR: XH NMR (400 MHz, Chloroforms/) 8 4.91 (d, J= 8.9 Hz, 1H), 3.94 - 3.81 (m, 1H), 3.68 (t, J= 6.1 Hz, 4H), 3.65 - 3.48 (m, 4H), 2.60 (t, J= 6.1 Hz, 4H), 1.42 (s, 9H). CNMR: 13C NMR (101 MHZ, cdch) 8 171.11, 155.31, 117.88, 79.70, 69.12, 65.53,
49.24, 28.30, 18.83, 14.16.
Figure imgf000284_0001
Compound 24 (4.96 g, 16.7 mmol) was dissolved in methanol (40 mL) and concentrated sulfuric acid (10 mL) was added. The mixture was heated at reflux for 24 hours under a nitrogen atmosphere, then cooled to room temperature. Excess sodium bicarbonate was then added, followed by di-tert-butyl dicarbonate (4.37 g, 20.04 mmol, 1.2 eq), and the reaction stirred at room temperature for 6 hours. The cloudy mixture was portioned between water and ethyl acetate, and the organic fraction washed with water (lx), ,5M HC1 (2x), saturated sodium bicarbonate (lx), and brine (lx), dried over sodium sulfate, and evaporated. Compound 20 was purified over a gradient of 0-10% MeOH in DCM on silica, and recovered in 74% yield (4.50 g, 12.4 mmol).
HNMR: XH NMR (400 MHz, Chloroforms/) 8 4.90 (d, J= 8.6 Hz, 1H), 3.82 (br s, 1H), 3.70 - 3.59 (m, 10H), 3.51 - 3.34 (m, 4H), 2.51 (t, J= 6.3 Hz, 4H), 1.38 (s, 9H).
CNMR: 13C NMR (101 MHz, cdch) 6 171.86, 155.34, 79.20, 69.19, 66.41, 51.58, 49.29, 34.75, 28.28.
Figure imgf000284_0002
TFA
Compound 25 (1.00 g, 2.75 mmol) was dissolved in dry MeOH (10 mL) and TFA (1 mL) and stirred for 15 minutes. Volatiles were evaporated under reduced pressure to give compound 26 as the TFA salt (1.04 g) in quantitative yield.
Figure imgf000285_0001
Compound 34 (50.0 mg, .123 mmol) was dissolved in DMF (5 mL) and DIPEA (214 uL, 159 mg, 1.23 mmol, 10 eq) and stirred under nitrogen. HBTU (102.4 mg, .270 mmol, 2.2 eq) was then added, and the reaction stirred for 15 minutes. Compound 26 (102 mg, .270 mmol, 2.2 eq) dissolved in DMF (1 mL) was then added dropwise, and the reaction stirred for 1 hour. The mixture was diluted into ethyl acetate, and washed with IM HC1 (2x) and brine (5x). The organic layer was evaporated to give a gummy residue, which was purified on reverse phase HPLC (35-45% MeCN in water, .1% TFA) to give compound 40 (62.6 mg, .0959 mmol) in 78% yield.
HRMS: expected 654.258, found 654.259
Compound 40 (62.6 mg, .0959 mmol) was dissolved in dioxane (1.8 mL) and IM NaOH (.2 mL) was added. The solution was stirred at room temperature for 2 hours, then acidified (pH 3) and evaporated. The residue was resuspended in EtOAc, washed with IM HC1, and dried over sodium sulfate. The organic layer was evaporated to give compound 41 as an oil (57.6 mg, .0921 mmol) in 96% yield, which was used without further purification.
Example 2-11. MIE binding trivalent (Figure 23)
Figure imgf000285_0002
Compound 34 (50.0 mg, .123 mmol) was dissolved in DMF (5 mL) and DIPEA (214 uL, 159 mg, 1.23 mmol, 10 eq) and stirred under nitrogen. HBTU (154 mg, .405 mmol, 3.3 eq) was then added, and the reaction stirred for 15 minutes. Compound 29 (200 mg, .405 mmol, 3.3 eq) dissolved in DMF (1 mL) was then added dropwise, and the reaction stirred for 1 hour. The mixture was diluted into ethyl acetate, and washed with IM HC1 (2x) and brine (5x). The organic layer was evaporated to give a gummy residue, which was purified on reverse phase HPLC (35-50% MeCN in water, .1% TFA) to give compound 44 (79.3 mg, .103 mmol) in 84% yield.
HRMS: [M+H]+ expected 770.305, found 770.308
Compound 44 (79.3 mg, .103 mmol) was dissolved in dioxane (1.8 mL) and IM NaOH (.2 mL) was added. The solution was stirred at room temperature for 2 hours, then acidified (pH 3) and evaporated. The residue was resuspended in EtOAc, washed with IM HC1, and dried over sodium sulfate. The organic layer was evaporated to give compound 41 as an oil (68.9 mg, .0948 mmol) in 92% yield, which was used without further purification.
Example 2-12. MIF-AcF3-2 (Figure 24)
Figure imgf000286_0001
Compound 41 (57.6 mg, .0921 mmol) was dissolved in DMF (1.8 mL) and DIPEA (.2 mL). HBTU (84.0 mg, .222 mmol, 2.4 eq) was then added, and the reaction stirred for 15 minutes before the addition of compound 15 (111 mg, .222 mmol, 2.4 eq). The reaction was stirred for 1 hour, then evaporated to give a red residue which was used in the next reaction without purification.
Compound 42 (crude, .0921 mmol scale) was dissolved in IM HC1 (1 mL) and stirred for 2 hours. The reaction was purified directly by HPLC (20-40% MeCN in H2O, +3% TFA) to give compound 43 (44.66 mg, .0295 mmol) in 32% yield.
Expected (H) 1514.570, found 1514.561
Example 2-13. MIF-AcF3-3 (Figure 25)
Figure imgf000287_0001
Compound 45 (68.9 mg, .0948 mmol) was dissolved in DMF (1.8 mL) and DIPEA (.2 mL). HBTU (126 mg, .333 mmol, 3.6 eq) was then added, and the reaction stirred for 15 minutes before the addition of compound 15 (167 mg, .333 mmol, 3.6 eq). The reaction was stirred for 1 hour, then evaporated to give a reddish residue which was used in the next reaction without purification.
Compound 38 (crude, .0948 mmol scale) was dissolved in IM HC1 (1 mL) and stirred for 2 hours. The reaction was purified directly by HPLC (20-40% MeCN in H2O, +3% TFA) to give compound 39 (78.1 mg, .0379 mmol) in 40% yield.
Expected (2H, /2) 1030.891 Found 1030.903
Example 2-14. Synthesis of MIF-AcF2-3, MIF-Ac-3, MIF-Et-3
Set forth in figures 26-29 are the chemical syntheses of MIF-AcF2-3, MIF-Ac-3, MIF-Et-3 and MIF-EtF3-3 produced using analogous methods to those presented above with minor variation.
The foregoing outlines features of several embodiments so that those skilled in the art may better understand the aspects of the present disclosure. Those skilled in the art should appreciate that they may readily use the present disclosure as a basis for designing or modifying other processes and structures for carrying out the same purposes and/or achieving the same advantages of the embodiments introduced herein. Those skilled in the art should also realize that such equivalent constructions do not depart from the spirit and scope of the present disclosure, and that they may make various changes, substitutions, and alterations herein without departing from the spirit and scope of the present disclosure.

Claims

CLAIMS What is claimed is:
1. A bifunctional compound, or a salt, stereoisomer, or solvate thereof, according to the chemical structure:
Figure imgf000288_0001
wherein:
[MIFBM/IgGBM] is a MIF or IgG binding moiety which binds respectively to circulating MIF or IgG, each of which is related to a disease state and/or condition and is to be removed by the action of hepatocytes on circulating protein;
[ASGPRBM] is a binding moiety which binds to hepatocytes through asialoglycoprotein receptors which are on the surface of hepatocytes, preferably in a patient or subject;
Each [CON] is an optional connector chemical moiety which, when present, connects directly to [MIFBM/IgGBM] or to [ASGPRBM] or connects the [LINKER] to [MIFBM/IgGBM] or to [ASGPRBM] and
[LINKER] is a chemical moiety having a valency from 1 to 15, often 1 to 10, more often 1 to 5 or 1, 2 or 3, which covalently attaches to one or more [ASGPRM] and/or [MIFBM/IgGBM] group, optionally through a [CON], including a [MULTICON] group, wherein said linker optionally itself contains one or more [CON] or [MULTICON] group(s); k’ is 1 to 15, 1 to 10, 1 to 5, 1 to 3 or 1, 2 or 3; j’ is 1 to 15, 1 to 10, 1 to 5, 1 to 3 or 1, 2 or 3; h and h’ are each independently 0 to 15;
IL is 0 to 15, often 1 to 15, 1 to 10, 1 to 5, 1 to 3, or 1, 2 or 3, preferably 11 is 1 to 5 or I, 2 or 3 with the proviso that at least one of h, h’ and 11 is preferably at least I, wherein the compound is selected from the group consisting of:
Figure imgf000289_0001
Figure imgf000290_0002
Figure imgf000290_0001
Figure imgf000291_0001
ein d
Figure imgf000291_0002
Figure imgf000291_0004
(¥01),
Figure imgf000291_0003
Figure imgf000292_0001
Figure imgf000293_0001
Figure imgf000294_0001
Figure imgf000295_0001
Figure imgf000296_0001
Figure imgf000297_0001
Figure imgf000298_0001
Figure imgf000299_0001
Figure imgf000300_0001
Figure imgf000301_0001
OOE
Figure imgf000302_0001
Figure imgf000303_0001
wherein: Extracellular Protein Targeting Ligand is a MIF or IgG Binding Moiety, which binds respectively to circulating MIF or IgG in a subject, each of which is related to a disease state or condition and is to be removed by action of hepatocytes or other cells of the subject;
Figure imgf000303_0002
Figure imgf000303_0003
(iv » hydrogen, R!v, alky!-C(())-R", -C(O)-R\ alkyl baloalkyl, -OC(O)R\ and -NRs-CtO)R.K’:
RK* Is selected front aryl alkyd-N'RMXOpR5, alkyl-atyl, alkyio'eteroaryi with L 2. or 4 heteroatoras, alkyLcyano, alkyl -OR”, alkyl~NR°R\ NRs-NRl'~C(0)Rl NR5-S(OBR\ alkenyl, allyl, alkyny I, ••N$Rf5-aIkenyi, -O- alkenyl, -NR''-alkynyk -NRMicteroaryl ••NR!'-aryl, -O-heteroaryl ■'O-aryL and -O-alkynyl, each of which R.!u is optionally substituted with 1, 2, 3, or 4 substituents;
R1 and R' are independently selected from hydrogen, beteroalkyl, Cfs-C$alkyl-cyauo, alkyl, alkenyl, alkynyl, haloalkyl, F, Cl, Br, 1, aryl, atyialkyl, heteroaryl, heteroarylalkyl, heterocycle, heterocycloalkyi, haloalkoxy, -O-alkenyl ~O~alkynyl, CBBlkyl- OR”, Cs-Gjalkyl-SR6. C^Ckalkyl-NR^R7, C(J-G,alkyl-Ct0)R\. Cs-Csalkyl-StOJRh. Ce-GBkyl- C(S)R\ C^Csalkyi-StGhB, CsBBkyi-N(R*)B(Q}R\ Cs-C4alkyl-N(RVS(O)R-’, Cn-G:;alkyl N(.Rs)-C(S}R< Cc-CcfdkyPN(Rs)-S(O)>R5 C;B?<alkyl-O-C(())R\ C;r€6aikyRO-S(O)R'\ Co- Csalkyl-0-C( S)Ry -N:::SfQ)(R '% Qj-Csalkylhb, and Cs-Csalkyl-G-StOjiR’, each of which Is optionally substituted with 1 . 2, 3, or 4 substituents;
R-’ at each occurrence is independently selected from hydrogen, alkyl, beteroalkyl haloalkyl (itidtnimg -CF?, -(RIB, -CH?F, -CHBFs -ClbCl feB and -CBCF-), arylalkyl heteroarylalkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocycle, -OR\ and -NRSR2
R4 is independently selected at each occurrence from hydrogen, heteroalkyl, allyl, haloalkyl, arylalkyl, heteroatylalkyl, alkenyl, alkynyl, aryl, heteroaryi, heterocycle, -ORR -N.R2R5, C(O)R\ S(O)R3, CCS)R\ and S(O)<R3;
IC and R1’ are independently selected at each occurrence from hydrogen, heteroalkyl, alkyl, atylalkyl. heteroarylalkyl, alkenyl, alkynyl, aryl, haloalkyl heteroaryi heterocycle, -alkyl-OR*, a.ikyl-NRsR5, C(O)R\ S(O)R\ C(S)R\ and S(OhR\
Rh and R;’ are independently selected at each occurrence from hydrogen, heteroalkyl alkyl aiyialkyi, heteroarylalkyl, alkenyl, alkynyl, atyl. heteroatyl, and heterocycle;
Cycle is a 3-8 mentbeted fused cyclic group optionally substituted with 1, 2, 3, or 4 substituents, each Linker’* is a bond or a moiety that covalently links the .ASGP.R. ligand to Linker5";
Linker* is a bond or a moiety that covalently links Linker* to an Extracellular Protein 'Bruetj ng Ligand, Linker*’ is a chemical group that links each Linker* to the Extracellular Protein Targeting Ligand; and
Linker1’ is a chemical group that links each Linker* to the .Extracellular Protein Targeting Ligand; and wherein, when IL is NR6-alkenyl, ■-NR'>-oH<y’?yL-NRsLXO)R!:i, -NI<s~S(Oh-alke-nyl, -NR8'-S(O).?“aIkynyI, -JS^Mieteroaryl, or -Nir-aryl., then Extracellular Protein Targeting Ligand dues not comprise an. oligonucleotide', and die optional substituents are selected from alkyl, alkenyl, alkynyl, haloalky L ~OR\ F, Cl, a
Br, I, ~NR' R?, heteroalkyi, cyano., nitro, C(O)R\
Figure imgf000305_0001
as allowed by valence such that a stable compound results.
2. The bifunctional compound of claim 1, which is selected from the group consisting of:
Figure imgf000305_0002
Figure imgf000306_0001
Figure imgf000307_0001
Figure imgf000308_0001
Figure imgf000309_0001
Figure imgf000310_0001
Figure imgf000311_0001
Figure imgf000312_0001
3. The bifunctional compound of claim 1, which is selected from the group consisting of:
Figure imgf000312_0002
Figure imgf000313_0001
Figure imgf000314_0001
Figure imgf000315_0001
The bifunctional compound of any one of claims 1-3, wherein R1 is H.
5. The bifunctional compound of any one of claims 1-4, wherein R2 is R10.
The bifunctional compound of any one of claims 1-5, wherein R10 is -NR6 -heteroaryl.
7. The bifunctional compound of any one of claims 1-6, wherein R6 is H.
8. The bifunctional compound of any one of claims 1-7, wherein -NR6 -heteroaryl is selected from the group consisting of:
Figure imgf000315_0002
9. The bifunctional compound of any one of claims 1-8, wherein the Extracellular Protein Targeting Ligand is a MIF binding moiety.
10. The bifunctional compound of any one of claims 1-9, wherein the Extracellular Protein Targeting Ligand is an IgG binding moiety.
11. The bifunctional compound of any one of claims 1-10, wherein Linked is selected from the group consisting of
Figure imgf000316_0001
12. A pharmaceutical composition comprising a therapeutically effective amount of a bifunctional compound of any one of claims 1-11, in combination with a pharmaceutically acceptable carrier, additive, or excipient, optionally further comprising an additional bioactive agent effective to treat, ameliorate, and/or prevent cancer, autoimmune disease, or inflammatory disease in the patient or subject or which is associated with the upregulation of MIF or IgG in the patient or subject.
13. The composition of claim 12, wherein the additional anticancer agent is selected from the group consisting of: everolimus, trabectedin, abraxane, TLK 286, AV-299, DN-101, pazopanib, GSK690693, RTA 744, ON ()9I0.Na, AZD 6244 (ARRY-142886), AMN-107, TKI-258, GSK461364, AZD 1152, enzastaurin, vandetanib, ARQ-197, MK-0457, MLN8054, PHA-739358, R-763, AT-9263, a FLT-3 inhibitor, a VEGFR inhibitor, an EGFR TK inhibitor, an aurora kinase inhibitor, a PIK-1 modulator, a Bcl-2 inhibitor, an HDAC inhbitor, a c-MET inhibitor, a PARP inhibitor, a Cdk inhibitor, an EGFR TK inhibitor, an IGFR-TK inhibitor, an anti-HGF antibody, a PI3 kinase inhibitors, an AK'T inhibitor, a JAK/STAT inhibitor, a checkpoint-! or 2 inhibitor, a focal adhesion kinase inhibitor, a Map kinase kinase (mek) inhibitor, a VEGF trap antibody, pemetrexed, erlotinib, dasatanib, nilotinib, decatanib, panitumumab, amrubicin, oregovomab, Lep-etu, nolatrexed, azd2171, batabulin, ofatumumab (Arzerra), zanolimumab, edotecarin, tetrandrine, rubitecan, tesmilifene, oblimersen, ticiliniumab, ipilimumab, gossypol, Bio 111, 131-I-TM-601 , ALT- 110, BIO 140, CC 8490, cilengitide, gimatecan, IL13-PE38QQR, INO 1001 , IPdRi KRX-0402, lucanthone, LY
317615, neuradiab, vitespan, Rta 744, Sdx 102, talampanel, atrasentan, Xr 311 , romidepsin, ADS- 100380. suniiinib. 5-fluorouracil, vorinostat. etoposide, gemcitabine, doxorubicin, irinotecan, liposomal doxorubicin, 5’-deoxy-5-fluorouridine, vincristine, temozolomide, ZK- 304709, seliciclib; PD0325901, AZD-6244, capecitabine, L-Glutamic acid, N -[4-[2-(2- amino-4,7“dihydro-4-oxo-l H - pyrroio[2,3- d ]pyrimidin-5-yl)ethyl]benzoyl]-, disodium salt, heptahydrate, camptothecin, PEG-labeled irinotecan, tamoxifen, toremifene citrate, anastrazole, exemestane, letrozole, DES(diethy I stilbestrol), estradiol, estrogen, conjugated estrogen, bevacizumab, IMC-1C11 , CHIR-258,); 3~[5-(methylsulfonylpiperadinemethyl)- indolylj -quinol one, vatalanib, /\G-013736, AVE-0005, the acetate salt of [D- Ser(Bu t ) 6 ,Azgly 10] (pyro-Glu-His-Trp-Ser-Tyr-D-Ser(Bu t )-Leu- Arg-Pro- Azgly-NH ?. acetate [CssIfeNisOi.; -(CsHtChjx where x ::: 1 to 2.4], goserelin acetate, leuprolide acetate, triptorelin pamoate, medroxyprogesterone acetate, hydroxyprogesterone caproate, megestrol acetate, raloxifene, bicalutamide, flutamide, nilutamide, megestrol acetate, CP-724714; TAK-
165, HK1-272, erlotinib, lapatanib, canertinib, ABX-EGF antibody, erbitux, EKB-569, PK I-
166, GW-572016, lonafamib, BMS-214662, tipifarnib; amifostine, NVP-LAQ824, suberoyl analide hydroxamic acid, valproic acid, trichostatin A, FK-228, SU11248, sorafenib, KRN951 , aminoglutethimide, arnsacrine, anagrelide, L-asparaginase, Bacillus Calmette- Guerin (BCG) vaccine, bleomycin, buserelin, busulfan, carboplatin, carmustine, chlorambucil, cisplatin, cladribine, clodronate, cyproterone, cytarabine, dacarbazine, dactinomycin, daunorubicin, diethyl stilbestrol, epirubicin, fludarabine, fludrocortisone, fluoxymesterone, flutamide, gemcitabine, gleevac, hydroxyurea, idarubicin, ifosfamide, imatinib, leuprolide, levamisole, lomustine, mechlorethamine, melphalan, 6-mercaptopurine, mesna, methotrexate, mitomycin, mitotane, mitoxantrone, nilutamide, octreotide, oxaliplatin, pamidronate, pentostatin, plicamycin, porfimer, procarbazine, raltitrexed, rituximab, streptozocin, teniposide, testosterone, thalidomide, thioguanine, thiotepa, tretinoin, vindesine, 13-cis-retinoic acid, phenylalanine mustard, uracil mustard, estramu stine, altretamine, floxuridine, 5 -deooxy uridine, cytosine arabinoside, 6-mecaptopurine, deoxy coformy ci n, calcitriol, valrubicin, mithramycin, vinblastine, vinorelbine, topotecan, razoxin, marimastat, COL-3, neovastat, BMS-275291, squalamine, endostatin, SU5416, SU6668, EMD121974, interleu kin- 12, IM862, angiostatin, vitaxin, droloxifene, idoxyfene, spironolactone, finasteride, cimitidine, trastuzumab, denileukin diftitox,gefitinib, bortezimib, paclitaxel, irinotecan, topotecan, doxorubicin, docetaxel, vinorelbine, bevacizumab, erbitux, cremophor- free paclitaxel, epithilone B, BMS- 247550, BMS-310705, droloxifene, 4-hydroxytamoxifen, pipendoxifene, ERA- 923, arzoxifene, fulvestrant, acolbifene, lasofoxifene, idoxifene, TSE- 424, HMR- 3339, ZK186619, PTK787/ZK 222584, VX-745, PD 184352, rapamycin, 40-0- (2-hydroxyethyl)-rapamycin, temsirolimus, AP -23573, RAD001, ABT-578, BC-210, LY294002, LY292223, LY292696, LY293684, LY293646, wortmannin, ZM336372, L- 779,450, PEG-filgrastim, darbepoetin, erythropoietin, granulocyte colony-stimulating factor, zolendronate, prednisone, cetuximab, granulocyte macrophage colony-stimulating factor, histrelin, pegylated interferon alfa-2a, interferon alfa-2a, pegylated interferon alfa-2b, interferon alfa-2b, azacitidine, PEG-L-asparaginase, lenalidomide, gemtuzumab, hydrocortisone, interleukin-11, dexrazoxane, alemtuzumab, all-transretinoic acid, ketoconazole, interleukin-2, megestrol, immune globulin, nitrogen mustard, methylprednisolone, ibritgumomab tiuxetan, androgens, decitabine, hexamethylmelamine, bexarotene, tositumomab, arsenic trioxide, cortisone, editronate, mitotane, cyclosporine, liposomal daunorubicin, Edwina-asparaginase, strontium 89, casopitant, netupitant, an NK-1 receptor antagonists, palonosetron, aprepitant, diphenhydramine, hydroxyzine, metoclopramide, lorazepam, alprazolam, haloperidol, droperidol, dronabinol, dexamethasone, methylprednisolone, prochlorperazine, granisetron, ondansetron, dolasetron, tropisetron, pegfilgrastim, erythropoietin, epoetin alfa and darbepoetin alfa, vemurafenib, PD-L1 checkpoint inhibitor, PD-1 checkpoint inhibitor, or CTLA-4 checkpoint inhibitor.
14. A method of removing excess circulating MIF or IgG in a patient or subject, the method comprising administering to the patient or subject a therapeutically effective amount of a bifunctional compound of any one of claims 1-11.
15. A method of treating, ameliorating, and/or preventing a disease state or condition which is associated with the upregulation of MIF or IgG in a patient or subject, the method comprising administering to the patient or subject a therapeutically effective amount of a bifunctional compound of any one of claims 1-11.
16. The method of claim 15, wherein the disease state or condition is cancer, an autoimmune disease, or an inflammatory disease.
17. A method of treating, ameliorating, or preventing cancer, an autoimmune disease, or an inflammatory disease in a subject or patient, the method comprising administering to the subject or patient a therapeutically effective amount of a bifunctional compound of any one of claims 1-11.
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