WO2025199466A1 - Ligands ciblant le récepteur d'asialoglycoprotéine spécifique du foie, conjugués les comprenant, compositions et procédés d'utilisation associés - Google Patents

Ligands ciblant le récepteur d'asialoglycoprotéine spécifique du foie, conjugués les comprenant, compositions et procédés d'utilisation associés

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Publication number
WO2025199466A1
WO2025199466A1 PCT/US2025/020963 US2025020963W WO2025199466A1 WO 2025199466 A1 WO2025199466 A1 WO 2025199466A1 US 2025020963 W US2025020963 W US 2025020963W WO 2025199466 A1 WO2025199466 A1 WO 2025199466A1
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conjugate
inhibitor
galnac
pharmaceutically acceptable
subject
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WO2025199466A9 (fr
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Philip S. Low
Madduri SRINIVASARAO
Rajeswara Reddy MANNEM
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Purdue Research Foundation
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Purdue Research Foundation
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
    • A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
    • A61K49/0032—Methine dyes, e.g. cyanine dyes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/549—Sugars, nucleosides, nucleotides or nucleic acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A61K49/005—Fluorescence in vivo characterised by the carrier molecule carrying the fluorescent agent
    • A61K49/0054—Macromolecular compounds, i.e. oligomers, polymers, dendrimers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics

Definitions

  • the present disclosure relates to conjugates that target an asialoglycoprotein receptor, such as in the liver, and pharmaceutical compositions thereof.
  • the conjugates of the present disclosure comprise an active agent, such as a therapeutic agent or an imaging agent.
  • Non-alcoholic fatty liver disease is a spectrum of liver disease in which fat builds up in the liver of people who drink little or no alcohol. This causes inflammation of the liver and damage to the cells in the liver, which may lead to fibrosis, cirrhosis (scarring of the liver) and liver failure.
  • An ideal drug for NASH would be a drug that targets fat deposition and metabolic disorders.
  • Thyroid hormones (THs) are essential regulatory molecules for normal growth and development and for maintaining metabolic homeostasis.
  • GalNAc conjugate I has been widely used in siRNA and antisense oligonucleotide (ASO) therapies by several pharmaceutical companies.
  • a disadvantage of GalNAc conjugate I is that it clears from the liver within about 10 hours.
  • T3 Triiodothryonine (T3) administration has a therapeutic effect on non-alcoholic steatohepatitis (NASH) and significantly eliminates hepatic steatosis. T3, however, causes heart burn and reduces bone mineral density. Therefore, there is a current need to provide conjugates which provide an advantage over currently available conjugates. This and other objects and advantages, as well as inventive features, will be apparent from the detailed description provided herein.
  • NASH non-alcoholic steatohepatitis
  • the present disclosure provides a conjugate of formula I: or a pharmaceutically acceptable salt thereof, wherein L 1 is a linker, each L 2 is independently a linker comprising a rigid component, A is an active agent, and each R is N-acetylgalactosamine (GalNAc), wherein the rigid component of L 2 comprises a structure of formula Ia: wherein: each X is independently -C(R 1 )2- or -NR 2 - , n is an integer selected from 2-7, each m is independently 0, 1, 2, 3, or 4, provided that if m is 0, then X is -C(R 1 ) 2 -, each R 1 is independently H, OH, or amino, and each R 2 is independently H or alkyl.
  • each X is independently -C(R 1 )2- or -NR 2 -
  • n is an integer selected from 2-7
  • each m is independently 0, 1, 2, 3, or 4, provided that if m is 0, then X is -C(R 1 )
  • the present disclosure provides a conjugate of formula I: formula (I) wherein L1 is a linker, L2 is a linker comprising a rigid component, A is an active agent, and R is a ligand that binds an asialoglycoprotein receptor (ASGPR).
  • the active agent can be a therapeutic agent, such as a therapeutic agent that has a therapeutic effect in the liver.
  • the therapeutic agent can be an imaging agent, such as a fluorescent imaging agent or a radio-imaging agent.
  • a pharmaceutical composition comprising an above-described conjugate and a pharmaceutically acceptable carrier. Further provided is a method of delivering an active agent to a liver in a subject.
  • the method comprises administering to the subject an effective amount of a conjugate as described herein, or a pharmaceutical composition comprising same and a pharmaceutically acceptable carrier.
  • the subject can have nonalcoholic fatty liver disease (NASH).
  • NASH nonalcoholic fatty liver disease
  • a method of imaging a liver in a subject comprises administering to the subject a conjugate as described herein (e.g., GalNAc-III-NIR (GalNAc conjugate III-56 containing S0456) or GalNAc-IV-NIR), or a pharmaceutical composition comprising same and a pharmaceutically acceptable carrier, and imaging the subject.
  • a method of treating a liver in a subject is administered to the subject an effective amount of a conjugate as described herein, or a pharmaceutical composition comprising same and a pharmaceutically acceptable carrier.
  • the method comprises administering to the subject an effective amount of a conjugate as described herein, or a pharmaceutical composition comprising same and a pharmaceutically acceptable carrier.
  • the subject can have nonalcoholic fatty liver disease (NASH), in which case the method can comprise administering to the subject an effective amount of a conjugate as described herein (e.g., GalNAc-III-T3 (GalNAc conjugate III-64) or GalNAc-IV-T3), or a pharmaceutical composition comprising same and a pharmaceutically acceptable carrier.
  • NASH nonalcoholic fatty liver disease
  • a method of improving the residence time in a liver of a mammal of a conjugate comprising an active agent A linked via a linker L to at least one ligand R that binds an asialoglycoprotein receptor (ASGPR) on a hepatocyte in the mammalian liver comprises modifying/replacing L with a linker having the structure L 1 (NH)C(CH 2 OL 2 R) x , wherein L 1 is a linker bound to A, L 2 is a linker comprising a rigid component, and x is an integer from 1 to 3.
  • the rigid component of L2 can be an oligoproline.
  • the rigid component of L 2 can be an oligopiperidine.
  • the rigid component of L 2 can be an ⁇ -helix.
  • the rigid component of L 2 can be peptidic.
  • the rigid component of L 2 can be the amino acid sequence (EAAAK)3.
  • the rigid component can be selected from: .
  • L2 can comprise an alkyl group, a PEG, a peptide, or a combination of two or more thereof.
  • the residence time in the liver of the mammal is at least three days.
  • a conjugate comprising an active agent A linked via a linker L to at least one ligand R that binds an asialoglycoprotein receptor (ASGPR) on a hepatocyte in a liver of a mammal
  • L has the structure L 1 (NH)C(CH 2 OL 2 R) x , wherein L1 is a linker bound to A, L2 is a linker comprising a rigid component, and x is an integer from 1 to 3, wherein the rigid component of L2 is an oligoproline, an oligopiperidine, an ⁇ -helix, the amino acid sequence (EAAAK)3, or a structure selected from: wherein L2 optionally further comprises an alkyl group, a PEG, a peptide, or a combination of two or more thereof; and wherein the conjugate has a residence time in the liver of the mammal of at least three days.
  • ASGPR asialoglycoprotein receptor
  • FIGURES Fig.1 shows the structure of GalNAc-III-54 (GalNAc-III-Tris-1), a precursor to a GalNAc conjugate III.
  • Fig.2 shows the structure of comparative GalNAc-I-NIR (GalNAc conjugate I containing S0456).
  • Fig.3 shows the structure of a GalNAc conjugate III, GalNAcIII-NIR (GalNAc conjugate III-56 containing S0456.
  • Fig.4 shows the structure of a GalNAc conjugate III (GalNAc conjugate III-56 without active agent (imaging agent, such as S0456, or therapeutic agent, such as T3)).
  • Figs.5A and 5B show images of Balb/c mice (10 weeks) (Fig.5A) and their excised organs (Fig.5B) 12 hours after being injected in the tail vein with either GalNAc-III-NIR (GalNAc conjugate III-56 containing S0456) alone or in the presence of GalNAc conjugate III-56 without S0456.
  • Figs.6A-6G show biodistribution images of excised organs of mice taken at 6 hours (Fig.6A), 10 hours (Fig.6B), 24 hours (Fig.6C), 48 hours (Fig.6D), 72 hours (Fig.6E), 106 hours (Fig.6F) and 130 hours (Fig.6G) after being injected in the tail vein with either GalNAc-I-NIR (GalNAc conjugate I containing S0456) (“I”) or GalNAc-III-NIR (GalNAc conjugate III containing S0456) (“III”).
  • Fig.7 shows the structure of GalNAc-III-T3, a GalNAc conjugate III containing T3.
  • Fig.8 shows the receptor saturation curve (concentration (nmol) vs.
  • Figs.9A and 9B show the pharmacokinetic (PK) stability of GalNAc-III-T3 administered intravenously (Fig.9A) and subcutaneously (Fig.9B).
  • Figs.10A and 10B show the stability of GalNAc-III-T3 (GalNAc-III-64) in human plasma over a period of 8 hours (Fig.10A) and 24 hours (Fig.10B).
  • Fig.14 shows aspartate aminotransferase (AST) levels (U/L) for mice fed a regular diet or a high fat diet (HFD) and then treated with either 10 nmol/day or 10 nmol/injection twice daily of GalNAc-III-T3.
  • Fig.15 shows alanine transaminase (ALT) levels (U/L) for mice fed a regular diet or a high fat diet (HFD) and then treated with either 10 nmol/day or 10 nmol/injection twice daily of GalNAc-III-T3.
  • Fig.16 shows triglyceride levels (mg/dl) for mice fed a regular diet or a high fat diet (HFD) and then treated with either 10 nmol/day or 10 nmol/injection twice daily of GalNAc- III-T3.
  • Fig.17 shows cholesterol levels (mg/dl) for mice fed a regular diet or a high fat diet (HFD) and then treated with either 10 nmol/day or 10 nmol/injection twice daily of GalNAc- III-T3.
  • HFD high fat diet
  • An object of the present disclosure is based on the discovery that rigid linkers that increase the residence time in the liver of conjugates targeted to aialoglycoprotein receptors (AGPRs).
  • L 1 is a linker
  • each L 2 is independently a linker comprising a rigid component
  • A is an active agent
  • each R is N-acetylgalactosamine (GalNAc)
  • the rigid component of L 2 comprises the structure of formula Ia: wherein: each X is independently -C(R 1 )2- or -NR 2 - , n is an integer selected from 2-7, each m is independently 0, 1, 2, 3, or 4, provided that if m is 0, then X is -C(R 1 ) 2 -, each R 1 is independently H, OH, or amino, and each R 2 is independently H or alkyl.
  • ASGPR asialoglycoprotein receptor
  • ASGPRs are lectins, which bind asialoglycoprotein and glycoproteins from which a sialic acid has been removed to expose a galactose residue. Integral membrane proteins, ASGPRs are locating on hepatocytes in mammals. ASGPRs are weakly expressed by glandular cells of the gallbladder and the stomach and are also expressed on several human carcinoma cell lines.
  • the active agent can be a therapeutic agent, such as a therapeutic agent that has a therapeutic effect in the liver.
  • the therapeutic agent can be a hormone.
  • the therapeutic agent can be a thyroid hormone receptor beta (THR ⁇ ) agonist, such as triiodothyronine (T3).
  • T3 thyroid hormone receptor beta
  • T3 triiodothyronine
  • the therapeutic agent can be a farnesoid X receptor (FXR) agonist, such as tropifexor (LJN452).
  • the therapeutic agent can be a peroxisome proliferator-activated receptor ⁇ agonist or ⁇ dual agonist; an example of an ⁇ dual agonist is elafibranor (GFT505).
  • the therapeutic agent can be an angiotensin II receptor blocker, such as telmisartan, candesartan, or losartan.
  • the therapeutic agent can be a patatin-like phospholipase domain-containing protein 3 (PNPLA3) inhibitor, such as an siRNA or an antisense oligonucleotide.
  • PNPLA3 patatin-like phospholipase domain-containing protein 3
  • the therapeutic agent can be a complement component C5 inhibitor, such as an siRNA or an antisense oligonucleotide.
  • the therapeutic agent can be a stearoyl-CoA desaturase-1 (SCD1) inhibitor, such as an siRNA or an antisense oligonucleotide.
  • SCD1 stearoyl-CoA desaturase-1
  • the therapeutic agent can be a diacylglycerol acyltransferase 1 (DGAT1) inhibitor, such as pradigastat (LCQ-908) or T-863.
  • DGAT1 diacylglycerol acyltransferase 1
  • the therapeutic agent can be a ⁇ -aminolevulinate synthase 1 inhibitor, such as an siRNA or an antisense oligonucleotide.
  • the therapeutic agent can be an angiopoietin-like 3 protein (ANGPTL3) inhibitor, such as an an siRNA or an antisense oligonucleotide.
  • ANGPTL3 angiopoietin-like 3 protein
  • the therapeutic agent can be a proprotein convertase subtilisin kexin type 9 (PCSK9) inhibitor, such as an siRNA or an antisense oligonucleotide.
  • PCSK9 proprotein convertase subtilisin kexin type 9
  • the therapeutic agent can be an apolipoprotein C-III (ApoC-III) inhibitor, such as an siRNA or an antisense oligonucleotide.
  • the therapeutic agent can be an apolipoprotein B (apoB) inhibitor, such as an siRNA or an antisense oligonucleotide.
  • the therapeutic agent can be a glycolate oxidase (GO) inhibitor, such as an siRNA or an antisense oligonucleotide.
  • GO glycolate oxidase
  • the therapeutic agent can be an ⁇ -1 antitrypsin (AAT) inhibitor, such as an siRNA or an antisense oligonucleotide.
  • the therapeutic agent can be a pyruvate kinase L/R (PKLR) inhibitor, such as an inhibitory polynucleotide (see, e.g., US 2020/0190523).
  • the therapeutic agent can be an antiviral small molecule drug, such as an antiviral small molecule drug for hepatitis A virus (HAV), hepatitis B virus (HBV), such as entecavir, tenofovir, lamivudine, adefovir, telbivudine, an A1CF inhibitor (see, e.g., US 2023/0257748), a Shwachman-Bodian-Diamond syndrome (SBDS) ribosome maturation factor inhibitor (see, e.g., US 2023/0193263), a Septin 9 (SEPT9) inhibitor (see, e.g., US 2023/0183692), a COPS signalosome subunit 3 (COPS3) inhibitor (see,
  • the therapeutic agent can be an antiviral siRNA or an antiviral antisense oligonucleotide (see, e.g., US 2022/0378920), such as an antiviral siRNA or an antiviral antisense oligonucleotide for HAV, HBV (see, e.g., US 2023/0357773, US 2023/0295630, US 2023/0119360, US 2021/0395745, US 2020/0171069, and US 2019/0343864), HCV, or HDV (see, e.g., US2021/0403908).
  • an antiviral siRNA or an antiviral antisense oligonucleotide see, e.g., US 2022/0378920
  • an antiviral siRNA or an antiviral antisense oligonucleotide for HAV, HBV see, e.g., US 2023/0357773, US 2023/0295630, US 2023/01
  • the therapeutic agent can be an siRNA or an antisense oligonucleotide (see, e.g., WO 2018/223073 (e.g., APOC3 oligonucleotide); WO 2018/223081 (e.g., PNPLA3 oligonucleotide); and Schmidt et al., Nucl Acids Res 45: 2294 (2017)).
  • the active agent can be an extracellular protein binding ligand for the selective degradation of a target extracellular protein to treat a disorder mediated by the extracellular protein (see, e.g., US 2024/0072809; and US 2023/0097256).
  • the therapeutic agent can be an imaging agent, such as a fluorescent imaging agent (fluorescent dye) or a radio-imaging agent.
  • the fluorescent dye can be carbocyanine, indocarbocyanine, oxacarbocyanine, thiacarbocyanine, merocyanine, polymethine, coumarine, rhodamine, xanthene, fluorescein, borondipyrromethane (BODIPY), CyS, CyS.S, Cy7, VivoTag-680, VivoTag-S680, VivoTag-S7S0, AlexaFluor660, AlexaFluor680, AlexaFluor700, AlexaFluor7S0, 10 AlexaFluor790, Dy677, Dy676, Dy682, Dy7S2, Dy780, DyLightS47, Dylight647, HiLyte Fluor 647, HiLyte Fluor 680, HiLyte Fluor 7
  • the fluorescent dye can have a peak emission in NIR-II window (1,000-1,700 nm).
  • the fluorescent dye can be selected from:
  • the imaging agent can comprise a chelating group, for example an imaging agent selected from DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid) or a derivative thereof; S-2-(4-isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane tetraacetic acid (p-SCN-Bn-DOTA) or derivative thereof; 2-S-(4-isothiocyanatobenzyl)-1,4,7- triazacyclononane-1,4,7-triacetic acid (p-SCN-Bn-NOTA) or a derivative thereof; [(R)-2- amino-3-(4-isothiocyanatophenyl)propyl]-trans-(S,S)-cyclohexane-1,2-diamine-pentaacetic acid (p-SCN-Bn-CHX-A”-DTPA) or a derivative thereof; TETA
  • the imaging agent can comprise a radioisotope/radionuclide for radio-imaging, radiotherapy, or magnetic resonance imaging (MRI).
  • the radioisotope/radionuclide can be selected from the group consisting of 177 Lu, 90 Y, 211 At, 225 Ac, 161 Tb, 18 F, 32 P, 44 Sc, 47 Sc, 52 Mn, 55 Co, 64 Cu, 6 7Cu, 67 Ga, 68 Ga, 86 Y, 89 Sr, 89 Zr, 99m Tc, 111 In, 114m In, 117m Sn, 124 I, 125 I, 131 I, 149 Tb, 153 Sm, 152 Tb, 155 Tb, 169 Er, 186 Re, 188 Re, 212 Pb, 212 Bi, 213 Bi, 223 Ra, 224 Ra, 225 Ab, and 227 Th.
  • the radioisotope/radionuclide can be selected from the group consisting of 11 C, 13 C, 13 N, 15 O, 60 Co, and 123 I.
  • the imaging agent can be or can comprise: wherein each X is independently a radioisotope selected from the group consisting of 18p, 124 I, 125 I, 131 I, and 2ll At; each R and R' is independently selected from -H, -D, -C1-C3 alkyl, benzyls, and substituted benzyls; and each n is independently an integer selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20.
  • the imaging agent can be or can comprise a radiolabeled prosthetic group selected from the group consisting of:
  • the imaging agent can comprise a chelating agent selected from:
  • the imaging agent can comprise a nuclide for positron emission tomography (PET) or single- photon emission computed tomography (SPECT).
  • the nuclide can be selected from the group consisting of 99m Tc, 111 In, 18 F, 68 Ga, 124 I, 125I , 131 I, and 64 Cu.
  • the linkers e.g., L 1 and each of L 2
  • the linker may have a wide variety of lengths, such as a range of atoms from about 7 to about 100.
  • the atoms used in forming the linker may be combined in all chemically relevant ways, such as chains of carbon atoms forming alkylene groups, chains of carbon and oxygen atoms forming polyoxyalkylene groups, chains of carbon and nitrogen atoms forming polyamines, and others.
  • the bonds connecting atoms in the chain may be either saturated or unsaturated, such that for example, alkanes, alkenes, alkynes, cycloalkanes, arylenes, imides, and the like may be divalent radicals that are included in the linker (e.g., L 1 and each of L 2 ).
  • the atoms forming the linker may also be cyclized upon each other to form divalent cyclic radicals in the linker(e.g., L 1 and each of L 2 ).
  • the chain forming the linker e.g., L 1 and each of L 2
  • the linker includes radicals that form one or more spacer linkers and/or releasable linkers that are taken together to form the linkers described herein having certain length, diameter, and/or functional group requirements.
  • releasable linkers may be used when the drug to be delivered is advantageously liberated from the binding ligand-linker conjugate so that the free therapeutic will have the same or nearly the same effect at the target as it would when administered without the targeting provided by the conjugates described herein.
  • the linker L is a non-releasable linker. It is appreciated that non-releasable linkers may be used when the active agent is advantageously retained by the conjugate, such as in imaging, diagnosing, uses of the conjugates described herein. It is to be understood that the choice of a releasable linker or a non-releasable linker may be made independently for each application or configuration of the conjugates, without limiting the invention described herein.
  • linkers L described herein comprise various atoms, chains of atoms, functional groups, and combinations of functional groups.
  • the linker L may be referred to by the presence of spacer linkers, releasable linkers, and heteroatoms.
  • L 1 can be a non-releasable linker.
  • L 1 can be a slowly releasable linker.
  • L 1 can comprise an alkyl group, a polyethylene glycol (PEG), a peptide, or a combination of two or more thereof.
  • PEG polyethylene glycol
  • L 1 can be a releasable linker.
  • L 1 can be reductively cleavable.
  • L 1 can be oxidatively cleavable.
  • L 1 can be enzymatically cleavable.
  • L 1 can be acid-cleavable.
  • L 1 is a releasable linker and comprises a disulfide bond.
  • L 1 can comprise an alkyl group (e.g., ethylene), a polyethylene glycol (PEG), a peptide, an aryl group (e.g., a phenyl or a phenoxy), a heteroaryl group, or a combination of two or more thereof.
  • L 1 comprises L 1 can comprise a portion having the structure: .
  • each L 2 is independently a linker comprising a rigid component.
  • each L 2 comprises a rigid component and preferably each L 2 comprises the same rigid component.
  • the rigid component of L 2 can improve residence time of the conjugate in the liver.
  • the rigid component of L 2 can include at least one heterocyclylene.
  • the prefix “oligo” can refer to 2 to 50 repeating components.
  • an "oligo amino acid,” or more precisely an “oligopeptide,” can refer to a short chain of amino acids (e.g., between 2 and 20 amino acid residues) joined by peptide (amide) bonds.
  • the rigid component of L 2 can be an oligoproline.
  • a proline can refer to a substituted or unsubstituted pyrrolidine-2-carboxylic acid (e.g., unsubstituted proline, hydroxy-proline, amino-proline, or methyl-proline).
  • the rigid component of L 2 can be an oligopiperidine.
  • a piperidine can refer to a substituted or unsubstituted piperidine-carboxylic acid (e.g., piperidine-2-carboxylic acid, piperidine -3-carboxylic acid, or piperidine -4-carboxylic acid).
  • the rigid component of L 2 can be an ⁇ -helix.
  • the rigid component of L 2 can be peptidic.
  • a peptidic rigid component of L 2 can comprise 2 to 50 amino acids (e.g., 2 to 7 amino acids).
  • the rigid component of L 2 can be the amino acid sequence (EAAAK)3.
  • the rigid component can be selected from: .
  • the rigid component of L 2 can comprise the structure of formula Ia: wherein: each X is independently -C(R 1 ) 2 - or -NR 2 - , n is an integer selected from 2-7, each m is independently 0, 1, 2, 3, or 4, provided that if m is 0, then X is -C(R 1 ) 2 -, each R 1 is independently H, OH, or amino, and each R 2 is independently H or alkyl.
  • X is -C(R 1 )2- (e.g., -CH2-).
  • X is -NR 2 - (e.g., -NH-).
  • n is 2, 3, 4, or 5.
  • n is 2, 3, or 4. In some embodiments, n is 2, 3, or 4. In some embodiments of formula Ia, each m is independently 1 or 2. In some embodiments, each m is 1. In some embodiments, each m is 2. In some embodiments of formula Ia, each R 1 is independently H, OH, or amino (e.g., NH2). In some embodiments, each R 1 is independently H or OH. In some embodiments, one R 1 is H and the other is OH. In some embodiments, each R 1 is H. In some embodiments of formula Ia, each R 2 is independently H or alkyl (e.g., C1-C6 alkyl, such as methyl). In some embodiments, R 2 is H.
  • each R 1 is independently H, OH, or amino (e.g., NH2). In some embodiments, each R 1 is independently H or OH. In some embodiments, one R 1 is H and the other is OH. In some embodiments, each R 1 is H. In some embodiments of formula Ia, each
  • Formula Ia can be selected from: Formula Ia can be selected from: Formula Ia can be selected from: .
  • Formula Ia can be selected from:
  • L 2 comprises a compound of formula wherein R, m, n, and X are as defined herein, A is a carbonyl, C 1 -C 6 alkylene, C 1 -C 6 alkylene-C(O)-, preferably is a carbonyl (e.g., to form an amide with the nitrogen-containing ring), wherein C 1 -C 6 alkyl is optionally substituted, (i) is an integer selected from 0 to 6, preferably is 2 or 3, (ii) is an integer selected from 0 to 6, preferably is 2 or 3, and B is amide (e.g., -NHC(O)-C1-C6 alkylene- such as -NHC(O)-ethylene-).
  • L 2 can comprise an alkyl group, a PEG, a peptide, or a combination of two or more thereof. Each of these groups can be linked together or to the rest of the molecule through various functional groups, including ethers, esters, amines, amides, carbonyls, and others.
  • the alkyl group, a PEG, a peptide, or a combination can be present between R and the rigid component, between the rigid component and , in some embodiments, L 2 comprises , , . conjugate can have a residence time in the liver of at least three days.
  • the conjugate can have a binding affinity (K D ) to asialoglycoprotein receptor of about 0.1 nM to about 100 nM.
  • R can be a carbohydrate.
  • Each R-L 2 can independently comprise the structure: , wherein X, n, and m are as described in formula Ia, and p is an integer of 1-12 (e.g., and integer of 1-8, such as 2, 3, 4, 5, or 6).
  • Each R-L 2 can independently comprise the structure: , wherein X, n, and m are as described in formula Ia.
  • Each R-L 2 can independently comprise the structure: , wherein n is as described in formula Ia.
  • Each R-L 2 can independently have the structure: H H , wherein n is as described in formula Ia.
  • Each R-L 2 can independently have the structure: .
  • the conjugate of formula I have the structure: , or a pharmaceutically acceptable salt thereof.
  • the conjugate of formula I (e.g., a GalNAc conjugate III) can have the structure: , or a pharmaceutically acceptable salt thereof.
  • the conjugate of formula I (e.g., a GalNAc conjugate IV) can have the structure: , or a pharmaceutically acceptable salt thereof.
  • the conjugate can have the structure: , or a pharmaceutically acceptable salt thereof.
  • the conjugate can have the structure: H HO or a pharmaceutically acceptable salt thereof.
  • the conjugate can have the structure: , or a pharmaceutically acceptable salt thereof.
  • the conjugate can have the structure:
  • a comparative conjugate without a rigid component such as a GalNAc conjugate I
  • a conjugate of formula I e.g., GalNAc conjugate III, such as GalNAc-III-NIR and GalNAc-III-T3
  • GalNAc conjugate III which has a rigid linker
  • a GalNAc conjugate I which does not have a rigid linker, clears from the liver within ten hours.
  • the conjugates of the present disclosure enable the delivery of an agent to hepatocytes, such as delivery of T3, while minimizing/avoiding off-target toxicities.
  • the conjugates can contain one or more asymmetric centers and thus give rise to enantiomers, diastereomers, and other stereoisomeric forms that are defined, in terms of absolute stereochemistry, as (R)- or (S)-. Unless stated otherwise, it is intended that all stereoisomeric forms of the conjugates are contemplated.
  • the conjugates described herein contain alkene double bonds, and unless specified otherwise, it is intended that this disclosure includes both E and Z geometric isomers (e.g., cis or trans).
  • geometric isomer refers to E or Z geometric isomers (e.g., cis or trans) of an alkene double bond.
  • positional isomer refers to structural isomers around a central ring, such as ortho-, meta-, and para- isomers around a benzene ring. Further, it is understood that replacement of one or more hydrogen atoms with deuterium can significantly lower the rate of metabolism of a drug and, therefore, increase its half-life.
  • substituted refers to a functional group in which one or more hydrogen atoms contained therein are replaced by one or more non-hydrogen atoms.
  • functional group or “substituent” as used herein refers to a group that can be or is substituted onto a molecule.
  • substituents or functional groups include, but are not limited to, a halogen (e.g., F, Cl, Br, and I); an oxygen atom in groups such as hydroxyl groups, alkoxy groups, aryloxy groups, aralkyloxy groups, oxo(carbonyl) groups, carboxyl groups including carboxylic acids, carboxylates, and carboxylate esters; a sulfur atom in groups such as thiol groups, alkyl and aryl sulfide groups, sulfoxide groups, sulfone groups, sulfonyl groups, and sulfonamide groups; a nitrogen atom in groups such as amines, azides, hydroxylamines, cyano, nitro groups, N-oxides, hydrazides, and enamines; and other heteroatoms in various other groups.
  • a halogen e.g., F, Cl, Br, and I
  • an oxygen atom in groups such as hydroxyl groups,
  • amino acid refers to an organic compound that contain both amino and carboxylic acid functional groups.
  • Amino acids can be classified according to the locations of the core structural functional groups (alpha- ( ⁇ -), beta- ( ⁇ -), gamma- ( ⁇ -) amino acids, etc.).
  • Alpha amino acids can be classified according to stereocenter of the alpha carbon, for example, in an L configuration or a D configuration.
  • An amino acid can refer to one of the 22 proteinogenic amino acids (e.g., alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, selenocysteine and pyrrolysine), or a non-proteinogenic amino acids (e.g., hydroxyproline, piperidine-carboxylic acid).
  • Alkyl generally refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, such as having from one to fifteen carbon atoms (e.g., C1-C15 alkyl). "Alkyl” is intended to include independent recitations of a saturated “alkyl, " unless otherwise stated. An alkyl can comprise one to thirteen carbon atoms (e.g., C1-C13 alkyl). An alkyl can comprise one to eight carbon atoms (e.g., C1-C8 alkyl). An alkyl can comprise one to five carbon atoms (e.g., C 1 -C 5 alkyl).
  • An alkyl can comprise one to four carbon atoms (e.g., C1-C4 alkyl).
  • An alkyl can comprise one to three carbon atoms (e.g., C1- C 3 alkyl).
  • An alkyl can comprise one to two carbon atoms (e.g., C 1 -C 2 alkyl).
  • An alkyl can comprise one carbon atom (e.g., C1 alkyl).
  • An alkyl can comprise five to fifteen carbon atoms (e.g., C 5 -C 15 alkyl).
  • An alkyl can comprise five to eight carbon atoms (e.g., C 5 -C 8 alkyl).
  • An alkyl can comprise two to five carbon atoms (e.g., C2-C5 alkyl).
  • An alkyl can comprise three to five carbon atoms (e.g., C 3 -C 5 alkyl).
  • the alkyl group is selected from methyl, ethyl, 1-propyl (n-propyl), 1-methylethyl (iso-propyl), 1-butyl (n-butyl), 1-methylpropyl (sec-butyl), 2-methylpropyl (iso-butyl), 1,1-dimethylethyl (tert-butyl), 1-pentyl (n-pentyl).
  • the alkyl is attached to the rest of the molecule by a single bond.
  • Alkoxy refers to a radical bonded through an oxygen atom of the formula –O-alkyl, where alkyl is an alkyl chain as defined above.
  • amide refers to a group wherein R 9 and R 10 each independently represent a hydrogen or hydrocarbyl group, or R 9 and R 10 taken together with the N atom to which they are attached complete a heterocycle having from 4 to 8 atoms in the ring structure.
  • amine and “amino” are art-recognized and refer to both unsubstituted and substituted amines and salts thereof, e.g., a moiety that can be represented by wherein R 9 , R 10 , and R 10 , each independently represent a hydrogen or a hydrocarbyl group, or R 9 and R 10 taken together with the N atom to which they are attached complete a heterocycle having from 4 to 8 atoms in the ring structure.
  • amino groups include, but are not limited to, -NH 2 , -NH(alkyl), -N(alkyl) 2.
  • carbonyl is art-recognized and refers to a group -C(O)- where a double bond exists between the carbon and oxygen.
  • esteer refers to a group —C(O)OR 9 wherein R 9 represents a hydrocarbyl group.
  • ether refers to a hydrocarbyl group linked through an oxygen to another hydrocarbyl group. Accordingly, an ether substituent of a hydrocarbyl group may be hydrocarbyl-O—. Ethers may be either symmetrical or unsymmetrical. Ethers include “alkoxyalkyl” groups, which may be represented by the general formula alkyl-O- alkyl.
  • Alkylene or “alkylene chain” generally refers to a straight or branched divalent alkyl group linking the rest of the molecule to a radical group, such as having from one to twelve carbon atoms, for example, methylene, ethylene, propylene, i-propylene, n-butylene, and the like.
  • Aryl refers to a radical derived from an aromatic monocyclic or multicyclic hydrocarbon ring system by removing a hydrogen atom from a ring carbon atom.
  • the aromatic monocyclic or multicyclic hydrocarbon ring system contains only hydrogen and from five to eighteen carbon atoms, where at least one of the rings in the ring system is fully unsaturated, i.e., it contains a cyclic, delocalized (4n+2) ⁇ –electron system in accordance with the Hückel theory.
  • the ring systems from which aryl groups are derived include, but are not limited to, benzene, fluorene, indane, indene, tetralin and naphthalene.
  • Alkyl or aryl-alkyl refers to a radical of the formula -R c -aryl, where R c is an alkylene chain as defined above, for example, methylene, ethylene, and the like. The alkylene chain part of the aralkyl radical is optionally substituted as described above for an alkylene chain.
  • Carbocyclyl or “cycloalkyl” refers to a stable non-aromatic monocyclic or polycyclic hydrocarbon radical consisting solely of carbon and hydrogen atoms, which includes fused or bridged ring systems, having from three to fifteen carbon atoms. A carbocyclyl can comprise three to ten carbon atoms.
  • a carbocyclyl can comprise five to seven carbon atoms.
  • the carbocyclyl is attached to the rest of the molecule by a single bond.
  • Carbocyclyl or cycloalkyl is saturated (i.e., containing single C-C bonds only) or unsaturated (i.e., containing one or more double bonds or triple bonds).
  • saturated cycloalkyls include, e.g., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.
  • An unsaturated carbocyclyl is also referred to as "cycloalkenyl.”
  • monocyclic cycloalkenyls include, e.g., cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl.
  • Polycyclic carbocyclyl radicals include, for example, adamantyl, norbornyl (i.e., bicyclo[2.2.1]heptanyl), norbornenyl, decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like.
  • Carbocyclylalkyl refers to a radical of the formula –R c -carbocyclyl, where R c is an alkylene chain as defined above.
  • Carboxy or “carboxylic acid group”, as used herein, refers to a group represented by the formula —CO 2 H.
  • Halo or “halogen” refers to a bromo, chloro, fluoro or iodo substituent.
  • Haloalkyl refers to an alkyl radical, as defined above, that is substituted by one or more halogen radicals, as defined above, for example, trifluoromethyl, difluoromethyl, fluoromethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, and the like.
  • heteroalkyl refers to an alkyl group as defined above in which one or more skeletal carbon atoms of the alkyl are substituted with a heteroatom (with the appropriate number of substituents or valencies – for example, -CH2- may be replaced with -NH- or -O-).
  • each substituted carbon atom is independently substituted with a heteroatom, such as wherein the carbon is substituted with a nitrogen, oxygen, selenium, or other suitable heteroatom.
  • a heteroalkyl is attached to the rest of the molecule at a carbon atom of the heteroalkyl.
  • a heteroalkyl is attached to the rest of the molecule at a heteroatom of the heteroalkyl.
  • a heteroalkyl is a C1-C18 heteroalkyl.
  • a heteroalkyl is a C 1 -C 12 heteroalkyl.
  • a heteroalkyl is a C 1 -C 6 heteroalkyl.
  • a heteroalkyl is a C 1 -C 4 heteroalkyl.
  • Heteroalkyl can include alkoxy, alkoxyalkyl, alkylamino, alkylaminoalkyl, aminoalkyl, heterocycloalkyl, heterocycloalkyl, and heterocycloalkylalkyl, as defined herein.
  • Heteroalkylene refers to a divalent heteroalkyl group defined above which links one part of the molecule to another part of the molecule.
  • Heterocyclyl refers to a stable 3- to 18-membered non-aromatic ring radical that can comprise two to twelve carbon atoms and from one to six heteroatoms selected from nitrogen, oxygen and sulfur. Unless stated otherwise specifically in the specification, the heterocyclyl radical is a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which optionally includes aromatic, fused, and/or bridged ring systems. The heteroatoms in the heterocyclyl radical are optionally oxidized. The heterocyclyl radical is partially or fully saturated. "Heterocyclyl” is intended to include independent recitations of heterocyclyl comprising aromatic and non-aromatic ring structures, unless otherwise stated.
  • heterocyclyl is attached to the rest of the molecule through any atom of the ring(s).
  • heterocyclyl radicals include, but are not limited to, dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl, 1,3-benzodioxolyl, 1,4-benzodioxanyl, tetrahydroquinolinyl, 5,6,7,8-tetrahydroquinazolinyl, 5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidinyl, 6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidinyl, 5,6,7,8-tetrahydropyrido[4,5-c]pyridazinyl, indolinyl, isoindolinyl, imidazolidinyl
  • a conjugate and/or composition can be administered directly into the blood stream, into muscle, or into an internal organ.
  • suitable routes for such parenteral administration include intravenous, intraarterial, intraperitoneal, intrathecal, epidural, intracerebroventricular, intraurethral, intrasternal, intracranial, intratumoral, intramuscular, intranasal, and subcutaneous.
  • Suitable means for parenteral administration include needle (including microneedle) injectors, needle-free injectors, and infusion techniques. Where it is desirable to deliver the compound(s) and/or compositions systemically, the compound(s) and/or composition can be formulated for parenteral administration by injection, e.g., by bolus injection or continuous infusion.
  • Formulations for injection can be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative.
  • the compositions can take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and can contain formulatory agents such as suspending, stabilizing and/or dispersing agents.
  • Parenteral formulations are typically aqueous or non-aqueous isotonic sterile solutions that can contain carriers or excipients, such as salts, carbohydrates, anti-oxidants, bactericide, solute and/or buffering agents (preferably at a pH of 3–9) which renders the composition isotonic with the blood of the intended subject, but, for some applications, they may be more suitably formulated as a sterile non-aqueous solution or as a dried form to be used in conjunction with a suitable vehicle, such as sterile, pyrogen-free water.
  • Such compositions can be presented in unit-dose or multi-dose sealed containers, for example, ampoules and vials.
  • a liquid formulation can be adapted for parenteral administration of a conjugate or composition as described herein.
  • the preparation of parenteral formulations under sterile conditions for example, by lyophilization under sterile conditions, can readily be accomplished using standard pharmaceutical techniques well-known to those skilled in the art.
  • the solubility of a conjugate can be increased by the use of appropriate formulation techniques, such as the incorporation of solubility-enhancing agents.
  • Formulations for parenteral administration can be formulated for immediate and/or modified release.
  • a conjugate can be administered in a time-release formulation, for example in a composition which includes a slow-release polymer.
  • the conjugate can be prepared with a carrier that will protect it against rapid release, such as a controlled release formulation, including implants and microencapsulated delivery systems.
  • Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, polylactic acid and polylactic, polyglycolic copolymers (PGLA). Methods for the preparation of such formulations are generally known to those skilled in the art.
  • Sterile injectable solutions can be prepared by incorporating the conjugate(s), alone or in further combination with one or more other active agents, in the required amount in an appropriate solvent with one or a combination of ingredients described above, as required, followed by filtered sterilization.
  • dispersions are prepared by incorporating the conjugate(s) into a sterile vehicle, which contains a dispersion medium and any additional ingredients of those described above.
  • the preferred methods of preparation are vacuum-drying and freeze-drying, which yield a powder of the active ingredients plus any additional desired ingredient from a previously sterile-filtered solution thereof, or the ingredients can be sterile- filtered together.
  • the pharmaceutical composition can be formulated as a solution, microemulsion, liposome, or other ordered structure suitable to high drug concentration.
  • the carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof.
  • a conjugate, or a pharmaceutical composition comprising a conjugate can be continuously administered, where appropriate.
  • a method of imaging a liver in a subject comprises administering to the subject an above-described conjugate (e.g., GalNAc conjugate III-56 containing S0456), or a pharmaceutical composition comprising same and a pharmaceutically acceptable carrier, and imaging the subject.
  • an above-described conjugate e.g., GalNAc conjugate III-56 containing S0456
  • a pharmaceutical composition comprising same and a pharmaceutically acceptable carrier
  • the subject is a mammal. In certain embodiments, the subject is a human.
  • the imaging can be performed after administering the conjugate(s), or pharmaceutically acceptable salt, hydrate or solvate thereof, or pharmaceutical composition comprising same.
  • the imaging can be performed by any suitable means, such as fluorescent imaging or radio-imaging.
  • the method can further comprise assessing or monitoring the efficacy of treatment.
  • the conjugates and/or compositions can be used to monitor tumor or lesion growth and proliferation quantitatively in vivo.
  • a method of monitoring a progression of a cancer in a subject comprising administering a conjugate, a pharmaceutically acceptable salt, solvate, hydrate, or stereoisomer thereof, or a pharmaceutical composition comprising the conjugate or a pharmaceutically acceptable salt, solvate, hydrate, or stereoisomer thereof to a subject.
  • Such method can further comprise imaging the cancer of the subject.
  • the subject can be imaged periodically over the course of a therapeutic treatment, and a practitioner can then compare the images and/or otherwise quantify lesion or cancer growth to determine therapeutic efficacy (e.g., if there is a differential killing effect of the cancer cells over the course of the therapeutic treatment, or a relative increase in lesion size or cancer growth).
  • a method for determining a likelihood of success of a therapeutic treatment in a subject.
  • the method further comprises assessing or monitoring efficacy of a treatment administered to the subject.
  • the conjugates, pharmaceutically acceptable salts, hydrates, or solvates thereof, and/or pharmaceutical compositions comprising the conjugate(s) are administered for imaging, the conjugate(s) pharmaceutically acceptable salts, hydrates, or solvates thereof, and/or compositions can be administered by any suitable route including, for example, intravenously, intraperitoneally, subcutaneously, intracranially, intradermally, intramuscularly, intraocularly, intrathecally, intracerebrally, and intranasally.
  • Amounts of currently available contrast agents can be used as a guide in determining the amounts of the conjugate(s), pharmaceutically acceptable salts or hydrates thereof, and/or compositions to be used.
  • the methods hereof further comprise treating the subject, or having the subject treated, for cancer.
  • the method can further comprise administering an effective amount of a treatment for cancer (e.g., a second anti-cancer therapy) at a site where the conjugate accumulates.
  • the treatment can be any suitable treatment, such as surgery, radiotherapy, brachytherapy, photodynamic therapy, photothermal therapy, focal ablation therapy including cryoablation, focal laser ablation and high-frequency ultrasound ablation, chemotherapy, and immunotherapy.
  • the therapeutic regimen for the treatment of a disease state can be determined by a person skilled in the art and will typically depend on factors including, but not limited to, the type, size, stage and receptor status of a tumor (e.g., with cancer) in addition to the age, weight and general health of the subject.
  • Another determinative factor can be the risk of developing recurrent disease. For instance, for a subject identified as being at high risk or higher risk or developing recurrent disease, a more aggressive therapeutic regimen can be prescribed as compared to a subject who is deemed at a low or lower risk of developing recurrent disease.
  • a more aggressive therapeutic regimen can be prescribed as compared to a subject that has a less advanced stage of cancer.
  • a method of treating a liver in a subject comprises administering to the subject an effective amount of an above-described conjugate, or a pharmaceutical composition comprising same and a pharmaceutically acceptable carrier.
  • the subject can have nonalcoholic fatty liver disease (NASH), in which case the method can comprise administering to the subject an effective amount of GalNAc-III-T3 (GalNAc conjugate III-64), or a pharmaceutical composition comprising same and a pharmaceutically acceptable carrier.
  • NASH nonalcoholic fatty liver disease
  • treat refers to any and all uses which remedy a condition or symptom, or otherwise prevent, hinder, retard, abrogate or reverse the onset or progression of cancer or other undesirable symptoms in any way whatsoever.
  • treating does not necessarily imply that a subject is treated until total recovery or cure.
  • the treatment need not necessarily remedy, prevent, hinder, retard, abrogate or reverse all signs or symptoms, but can remedy, prevent, hinder, retard, abrogate or reverse one or more signs or symptoms.
  • therapeutically effective amount means the amount of conjugate when administered to a mammal, in particular a human, in need of such treatment, is sufficient to treat cancer.
  • the precise amount of conjugate to be administered can be determined by a physician with consideration of individual differences in age, weight, tumor size, extent of infection or metastasis, and condition of the subject.
  • administering of the conjugate(s), pharmaceutically acceptable salt, hydrate, or solvate thereof, and/or composition to a subject is meant that the conjugate(s), pharmaceutically acceptable salt, hydrate, or solvate thereof, or composition is presented such that the conjugate(s) and/or pharmaceutically acceptable salts, hydrates, or solvates thereof can be transferred to the subject.
  • the mode of administration will generally be by way of oral, parenteral (including subcutaneous, intradermal, intramuscular, intravenous, intracerebrally, intranasally, intrathecal, and intraspinal), inhalation (including nebulization), topical, rectal and vaginal modes.
  • the conjugate(s), pharmaceutically acceptable salt, hydrate, or solvate thereof, and/or composition can also be administered directly into a tumor and/or into tissue adjacent one or more segments of a tumor or administered directly into blood vessels.
  • the conjugate(s), pharmaceutically acceptable salt, hydrate, or solvate thereof, and/or composition can be administered in, as appropriate, a treatment or diagnostic effective amount.
  • a treatment or diagnostic effective amount includes an amount which, when administered according to the desired dosing regimen, achieves a desired therapeutic or diagnostic effect, including one or more of: alleviating the symptoms of, preventing or delaying the onset of, inhibiting or slowing the progression of, diagnosing, or halting or reversing altogether the onset or progression of a particular condition being treated and/or assessed.
  • effective amount means and encompasses both therapeutically effective amount and treatment or diagnostic effective amount. Suitable dosage amounts and dosing regimens to achieve this can be determined by the attending physician and can depend on the particular condition being treated or diagnosed, the severity of the condition as well the general age, health and weight of the subject. Depending upon the route of administration, a wide range of permissible dosages are contemplated.
  • Suitable dosages of the particulate material per se can lie within the range of about 0.1 ng per kg of body weight to 1 g per kg of body weight per dosage.
  • the dosage can be in the range of 1 ⁇ g to 1 g per kg of body weight per dosage, such as is in the range of 1 mg to 1 g per kg of body weight per dosage.
  • the dosage can be in the range of 1 mg to 500 mg per kg of body weight per dosage.
  • the dosage can be in the range of 1 mg to 250 mg per kg of body weight per dosage.
  • the dosage can be in the range of 1 mg to 100 mg per kg of body weight per dosage, such as up to 50 mg per body weight per dosage.
  • Conjugate(s), pharmaceutically acceptable salt, hydrate, or solvate thereof, and/or compositions hereof can be administered in a single dose or a series of doses.
  • dosages may be single or divided and may be administered according to a wide variety of protocols, including q.d. (once a day), b.i.d. (twice a day), t.i.d. (three times a day), or even every other day, once a week, once a month, once a quarter, and the like.
  • the effective amounts described herein correspond to the instance of administration, or alternatively to the total daily, weekly, month, or quarterly dose, as determined by the dosing protocol.
  • an effective amount of any one or a mixture of the compounds described herein can be determined by the attending diagnostician or physician by the use of known techniques and/or by observing results obtained under analogous circumstances.
  • a number of factors are considered by the attending diagnostician or physician, including, but not limited to the species of mammal, including human, its size, age, and general health, the specific disease or disorder involved, the degree of or involvement or the severity of the disease or disorder, the response of the individual patient, the particular compound administered, the mode of administration, the bioavailability characteristics of the preparation administered, the dose regimen selected, the use of concomitant medication, and other relevant circumstances.
  • a use of a conjugate, a pharmaceutically acceptable salt, hydrate, or solvate of the conjugate, or a composition hereof in the manufacture of a medicament for the treatment of a disease in a subject is provided.
  • the conjugate can be any compound or conjugate hereof.
  • the disease in the subject can be cancer.
  • the disease in the subject can be fibrosis.
  • the disease in the subject can be an inflammatory disease or disorder.
  • Any of the conjugates and/or compositions hereof can be for use in the treatment of a subject experiencing and/or having a disease state described herein.
  • the disease state for example, can be cancer, fibrosis, or an inflammatory disease or disorder.
  • connection between two components does not necessarily mean a direct, unimpeded connection, as a variety of other components may reside between the two components of note. Consequently, a connection does not necessarily mean a direct, unimpeded connection unless otherwise noted.
  • a method of improving the residence time in a liver of a mammal of a conjugate comprising an active agent A linked via a linker L to at least one ligand R that binds an asialoglycoprotein receptor (ASGPR) on a hepatocyte in the mammalian liver.
  • A active agent A linked via a linker L to at least one ligand R that binds an asialoglycoprotein receptor (ASGPR) on a hepatocyte in the mammalian liver.
  • ASGPR asialoglycoprotein receptor
  • the method comprises modifying/replacing L with a linker having the structure L 1 (NH)C(CH 2 OL 2 R) x , wherein L 1 is a linker bound to A, L 2 is a linker comprising a rigid component, and x is an integer from 1 to 3.
  • the rigid component of L2 can be an oligoproline.
  • the rigid component of L 2 can be an oligopiperidine.
  • the rigid component of L 2 can be an ⁇ -helix.
  • the rigid component of L 2 can be peptidic.
  • the rigid component of L 2 can be the amino acid sequence (EAAAK)3.
  • the rigid component can be selected from: .
  • L2 can comprise an alkyl group, a PEG, a peptide, or a combination of two or more thereof.
  • the residence time in the liver of the mammal is at least three days.
  • a conjugate comprising an active agent A linked via a linker L to at least one ligand R that binds an asialoglycoprotein receptor (ASGPR) on a hepatocyte in a liver of a mammal, wherein L has the structure L 1 (NH)C(CH 2 OL 2 R) x , wherein L1 is a linker bound to A, L2 is a linker comprising a rigid component, and x is an integer from 1 to 3, wherein the rigid component of L2 is an oligoproline, an oligopiperidine, an ⁇ -helix, the amino acid sequence (EAAAK)3, or a structure selected from: wherein L2 optionally further comprises an alkyl group, a PEG, a peptide, or a combination of two or more thereof; and wherein the conjugate has a residence time in the liver of the mammal of at least three days.
  • a conjugate of formula I formula (I) wherein L 1 is a linker, L 2 is a linker comprising a rigid component, A is an active agent, and R is a ligand that binds an asialoglycoprotein receptor (ASGPR).
  • A is an active agent
  • R is a ligand that binds an asialoglycoprotein receptor (ASGPR).
  • the therapeutic agent has a therapeutic effect in the liver.
  • the therapeutic agent is a hormone.
  • the therapeutic agent is a thyroid hormone receptor beta (THR ⁇ ) agonist.
  • THR ⁇ agonist is triiodothyronine (T3).
  • the conjugate of clause 2, wherein the therapeutic agent is a patatin-like phospholipase domain-containing protein 3 (PNPLA3) inhibitor.
  • PNPLA3 inhibitor is an siRNA or an antisense oligonucleotide.
  • the therapeutic agent is a complement component C5 inhibitor.
  • the complement component C5 inhibitor is an siRNA or an antisense oligonucleotide.
  • the therapeutic agent is a stearoyl-CoA desaturase-1 (SCD1) inhibitor.
  • SCD1 inhibitor is an siRNA or an antisense oligonucleotide. 20.
  • the ANGPTL3 inhibitor is an siRNA or an antisense oligonucleotide.
  • the therapeutic agent is a proprotein convertase subtilisin kexin type 9 (PCSK9) inhibitor.
  • PCSK9 inhibitor is an siRNA or an antisense oligonucleotide.
  • the therapeutic agent is an apolipoprotein C- III (ApoC-III) inhibitor.
  • the ApoC-III inhibitor is an siRNA or an antisense oligonucleotide.
  • the conjugate of clause 2, wherein the therapeutic agent is an apolipoprotein B (apoB) inhibitor.
  • apoB apolipoprotein B
  • the apoB inhibitor is an siRNA or an antisense oligonucleotide.
  • the therapeutic agent is a glycolate oxidase (GO) inhibitor.
  • the GO inhibitor is an siRNA or an antisense oligonucleotide.
  • the therapeutic agent is an ⁇ -1 antitrypsin (AAT) inhibitor.
  • AAT ⁇ -1 antitrypsin
  • the therapeutic agent is an antiviral small molecule drug.
  • the antiviral small molecule drug is an antiviral small molecule drugs for hepatitis A virus (HAV), hepatitis B virus (HBV), or hepatitis C virus (HCV).
  • HBV hepatitis A virus
  • HBV hepatitis B virus
  • HCV hepatitis C virus
  • 38. The conjugate of clause 37, wherein the antiviral small molecule drug for HBV is entecavir, tenofovir, lamivudine, adefovir, or telbivudine. 39.
  • R is a carbohydrate.
  • R is a monosaccharide.
  • R is selected from galactose, glucose, fucose, mannose, galactosamine, rhamnose, rhamnitol, N-acetylgalactosamine (GalNAc), mannosamine, glucosamine, N-acetyl-glucosamine (GluNAc), fucosamine and rhamnosamine.
  • a pharmaceutical composition comprising a conjugate of any one of clauses 1-69 and a pharmaceutically acceptable carrier.
  • 71. A method of imaging a liver in a subject, which method comprises administering to the subject a conjugate of any one of clauses 43-45 and 68, or a pharmaceutical composition comprising same and a pharmaceutically acceptable carrier, and imaging the subject.
  • 72. A method of treating a liver in a subject, which method comprises administering to the subject an effective amount of a conjugate of any one of clauses 1-42, 46-67, and 69, or a pharmaceutical composition comprising same and a pharmaceutically acceptable carrier, whereupon the liver in the subject is treated.
  • a method of improving the residence time in a liver of a mammal of a conjugate comprising an active agent A linked via a linker L to at least one ligand R that binds an asialoglycoprotein receptor (ASGPR) on a hepatocyte in the mammalian liver comprises modifying/replacing L with a linker having the structure L1(NH)C(CH2OL2R)x, wherein L 1 is a linker bound to A, L 2 is a linker comprising a rigid component, and x is an integer from 1 to 3.
  • the rigid component of L 2 is an oligoproline.
  • the method of clause 75, wherein the rigid component of L2 is an oligopiperidine. 78. The method of clause 75, wherein the rigid component of L2 is an ⁇ -helix. 79. The method of clause 75, wherein the rigid component of L2 is peptidic. 80. The method of clause 75, wherein the rigid component of L2 is the amino acid sequence (EAAAK)3. 81. The method of clause 75, wherein the rigid component is selected from: . 82. The method of clause 75, wherein L2 comprises an alkyl group, a PEG, a peptide, or a combination of two or more thereof. 83. The method of clause 75, wherein the residence time in the liver of the mammal is at least three days. 84.
  • a conjugate comprising an active agent A linked via a linker L to at least one ligand R that binds an asialoglycoprotein receptor (ASGPR) on a hepatocyte in a liver of a mammal, wherein L has the structure L 1 (NH)C(CH 2 OL 2 R) x , wherein L 1 is a linker bound to A, L2 is a linker comprising a rigid component, and x is an integer from 1 to 3, wherein the rigid component of L2 is an oligoproline, an oligopiperidine, an ⁇ -helix, the amino acid sequence (EAAAK)3, or a structure selected from:
  • L2 optionally further comprises an alkyl group, a PEG, a peptide, or a combination of two or more thereof; and wherein the conjugate has a residence time in the liver of the mammal of at least three days.
  • Step 2 Synthesis of Gal-2 Freshly prepared Gal-1 (1.5 g, 1.0 eq.) was dissolved in anhydrous 1,2-dichloroethane (8 vol) at room temperature, 4 ⁇ molecular sieve powder (1.0 eq, wt/wt) was added to the reaction, and the reaction was allowed to stir for 5 min at room temperature. 5-hexen-1-ol (1.2 eq.) was added under N2 atmosphere and allowed to stir for another 30 min. Finally, TMSOTf was added slowly, and stirring continued for 2 hours at room temperature. After the reaction was completed (monitored by LCMS), the reaction mixture was quenched with sat. NaHCO3 solution and filtered to remove the molecular sieves.
  • Step 2 Synthesis of Tris-2 Tris-1 was dissolved in EtOH . HCl (3.3 M, 10 mL/1.0 g) and allowed to stir at reflux temperature (80 o C an internal temperature). After completing the reaction (monitored by LCMS, usually 6 hours), the resulting solution was concentrated, and the resultant crude was diluted with DCM and washed with Sat. NaHCO 3 (expected to be exothermic) solution, followed by brine, and dried over Na2SO4. The crude compound was purified by flash chromatography (DCM:MeOH, 9:1) (Tris-2).
  • Step 3 Synthesis of Tris-3 To a stirred solution of Tris-2 (7.0 g,16.62 mmol) in dioxane was added Na 2 CO 3 (2.642 g, 24.93 mmol) followed by water (24 mL). To this solution was added Cbz-Cl (2.44 mL, 18.33 mmol), and the reaction was allowed to continue for 12 hours. The solvent was removed under reduced pressure, and the resulting oil was purified by flash chromatography (Hexanes: EtOAc, 1:1) to yield compound Tris-3 as a colorless liquid.
  • Step 4 Synthesis of Tris-4 To a stirred solution of Tris-3 (2.2 g, 3.96 mmol) in MeOH (30 mL) was added LiOH . H2O (1,42 g, 59.4 mmol) followed by water: THF (30 mL, 1:1). The resulting solution was allowed to stir for 1 hour at room temperature. After completion of reaction, pH was acidified to 2 with concentrated HCl. The volatile solvents were removed under reduced pressure using rotavapor, the resulting oil was diluted with EtOAc, and the organic layer was washed with water and dried over Na2SO4.
  • Step 5 Synthesis of Tris-5 To Tris-4 (1.5 g, 3.18 mmol) in DMF (20 mL) was added HBTU (5.42 g, 14.31 mmol), HOBt (1.93 g, 14.31, 4.5 eq.) and DIPEA (5.52 mL, 31.8 mmol, 10.0 eq.), and the resulting solution was allowed to stir for 15 min.
  • Step 6 Synthesis of Tris-6 To Tris-5 in DCM (10 vol) was added trifluoroacetic acid (3 vol), and the reaction mixture was allowed to stir at room temperature for 30 min. Boc deprotection was confirmed by LCMS, and the resulting solution was concentrated and co-distilled with toluene. The resultant compound was used for the next step without any further purification.
  • Step 7 Synthesis of GalNAc-Tris-1 To Gal-3 (2.31 g, 5.17 mmol) in DMF (20 mL) was added HBTU (2.52 g, 6.66 mmol), HOBt (0.89 g, 6.66 mmol, 4.5 eq.) and DIPEA (2.57 mL, 14.8 mmol, 10.0 eq.), and the resulting solution was allowed to stir for 15 min. To the resulting solution, Tris-6 (0.95 g, 1.48 mmol, 1.0 eq.) in DMF (6 mL) was added at room temperature and allowed to stir for 12 hours. After reaction completion, as monitored by TLC, the reaction mixture was diluted with 1 M phosphoric acid and extracted with DCM.
  • Step 8 Synthesis of GalNAc-Tris-2 To GalNAc-Tris-1 (0.760 g) in MeOH (10 mL) was added 10% Pd on carbon (0.08 g, 10% wt/wt), and the reaction mixture was allowed to stir at room temperature for 2 hours under hydrogen atmosphere (balloon pressure). Cbz deprotection was confirmed by TLC, and the reaction mixture was filtered through a celite pad.
  • Step-9 Synthesis of GalNAc-Tris-3
  • HATU 0.413 g, 1.087 mmol
  • DIPEA 1.3 mL, 7.25 mmol, 10.0 eq.
  • NIR-acid As a green solid. LCMS m/z calcd for C 47 H 58 N 2 O 15 S 4 (M + H) + 1017.26 found 1017.31.
  • Ac Ac Step-2 To NIR-acid (0.020 g, 0.02 mmol) in dry DMSO was added HATU (0.033 g, 0.09 mmol, 4.5 eq.), and DIPEA (0.04, 0.2 mmol, 10.0 eq.). The resulting solution was allowed for stirring for 15 min. To the solution, GalNAc-Tris-4 (0.04 g, 0.013 mmol) was added at rt and allowed to stir for 1h.
  • Example 4 Step 1 Synthesis of Gal-1 Commercially available galactosamine pentaacetate (8.0 g, 20.5 mmol, 1.0 eq.) was dissolved in anhydrous dichloroethane (50 mL) at room temperature, and TMSOTf (5.98 mL, 32.9 mmol, 1.6 eq.) was added to the reaction mixture. The reaction was stirred at 60 o C for 90 minutes, allowed to come to room temperature, and stirred for another 12 hours. The reaction was diluted with sat. NaHCO3 solution, and the product was extracted with dichloroethane. The combined organic layer was washed with water followed by brine solution and dried over Na 2 SO 4 .
  • Step 2 Synthesis of Gal-2 Freshly prepared Gal-1 (1.5 g, 1.0 eq.) was dissolved in anhydrous 1,2-dichloroethane (8 vol) at room temperature, 4 ⁇ molecular sieve powder (1.0 eq, wt/wt) was added to the reaction, and the reaction was allowed to stir for 5 min at room temperature. 3-buten-1-ol (1.2 eq.) was added under N 2 atmosphere and allowed to stir for another 30 min.
  • Step 3 Synthesis of GalNAc-III-8 Freshly prepared Gal-2 (7.0 g, 1.0 eq.) was dissolved in dichloroethane (3 vol) and acetonitrile (3 vol) at room temperature. To the resultant clear solution, sodium metaperiodate (13.96 g.4.0 eq) was added, and the mixture was allowed to stir for 5 min at room temperature. RuCl3.H2O (0.14 g, 0.05 eq.) was added to the reaction mixture, and the mixture was allowed to stir at 30 o C (highly exothermic, cold-water bath) for another hour. To the resultant reaction mixture, another 1.0 eq. of NaIO4 was added, and the mixture was allowed to stir for another 3 hours at room temperature.
  • reaction progress was monitored by LCMS.
  • the reaction mixture was diluted with water, and the pH of the resultant solution was adjusted to 7.5 with solid NaHCO 3 and filtered to remove the undissolved particles.
  • the filtrate was washed with dichloromethane.
  • the organic layer was discarded, and the aqueous layer was acidified to pH 2.0 with citric acid and saturated with NaCl.
  • the resultant solution was extracted with EtOAc (3 x 500 mL).
  • the resultant organic layer was dried over Na2SO4, and solvents were concentrated under reduced pressure.
  • the resultant crude was purified by combi-flash (MeOH:CH2Cl2, 1:9) to get GalNAc-III-8 (Gal-8) as a colorless viscous liquid.
  • Tris-9 Step-3 To Tris-8 (6.3 g, 14.4 mmol) in dry DMF (100 mL) were added HATU (24.68 g, 64.8 mmol, 4.5 eq.) and DIPEA (15.02 mL, 86.4 mL, 6.0 eq.), and the resulting solution was allowed to stir for 15 min. To the solution, NFmoc-1,2-ethylenediamine (24.68 g, 64.8 mmol) was added at room temperature, and the resulting solution was allowed to stir for 2 hours. After reaction completion, as monitored by LCMS, the mixture was diluted with DCM and washed with water.
  • Step-4 Synthesis of Tris-9 To the product of step 3 (3.2 g) in DCM (30 mL) was added trifluoroacetic acid (10 mL), and the reaction mixture was stirred at room temperature for 30 min. Boc deprotection was confirmed by LCMS, and the resulting solution was concentrated and co-distilled with toluene. The resultant compound was used for the next step without any further purification. LCMS m/z calc’d for C64H72N7O12 (M + H) + 1130.52; found 1130.87. Synthesis of Tris-10
  • Step-5 To Tris-9 (1.0 g, 0.884 mmol) in dry DMF (10 mL) were added PyBOP (0.919 g, 1.769 mmol) and DIPEA (0.307 mL, 1.769 mmol), and the resulting solution was allowed to stir for 15 min.4-(tert-butoxy)-4-oxobutanoic acid (0.310 g, 1.769 mmol) was added at room temperature, and the solution was stirred for two hours. After reaction completion, as monitored by LCMS, the mixture was diluted with DCM and washed with water. The resultant organic layer was concentrated and purified by combi-flash using a mixture of CH2Cl2 and MeOH as eluant.
  • Step-6 Synthesis of Tris-10 To the product of step 5 (3.0 g) in DCM (30 mL) was added trifluoroacetic acid (10 mL), and the reaction mixture was allowed to stir at room temperature for two hours. Tertiary butyl deprotection was confirmed by LCMS, and the resulting solution was concentrated and co-distilled with toluene. The resultant compound was used for the next step without any further purification.
  • Example 6 Synthesis of GalNAc-III-Tris-1 (GalNAc-III-54)
  • 1,2-diaminoethane trityl resin 1.0 g, 0.98 mmol was swollen with dichloromethane (20 mL) for 20 mins followed by dimethyl formamide (2 ⁇ 15 mL) for 15 mins each. Then, a solution of Tris-10 (2.41 g, 1.96 mmol), Cl-HOBt (0.331 g, 1.96 mmol), tripyrrolidinophosphonium hexafluorophosphate (PyBOP, 1.02 g, 1.96 mmol), and N, N- Diisopropylethylamine (DIPEA) (5.1 mL, 30 mmol) in DMF (30 mL) was added.
  • DIPEA Diisopropylethylamine
  • GalNAc-III-Tris-1 was cleaved from the resin using the cocktail solution DCM: TFA: triisopropylsilane (TIPS) (55:40:5) (3 ⁇ 20 mL) for 20 min each time. The compound was concentrated under vacuum. Excess TFA was evaporated under rotavapor. Then, the crude product was precipitated in cold diethyl ether solution and centrifuged at 2,000 rpm for 10 mins. The supernatant was removed.
  • Step-1 Synthesis of GalNAc-III-NIR-1 To NIR-acid (0.009 g, 0.008195 mmol), in dry DMSO were added HATU (0.012 g, 0.02983 mmol) and DIPEA (0.0129 mL, 0.0745 mmol), and the resulting solution was stirred for 15 min. To the solution, GalNAc-III-Tris-1 (0.02 g, 0.00745 mmol) was added at room temperature and allowed to stir for one hour. After reaction completion, as monitored by LCMS, the mixture was diluted with water. The product was purified by HPLC.
  • Step-2 Synthesis of GalNAc-III-NIR (GalNAc conjugate-III-56 containing S0456) To GalNAc-III-NIR-1 (0.005 g, 0.02 mmol) was added 10 % aq. NH3 (0.5 mL), and the mixture was stirred for 30 min.
  • Example 8 Synthesis of GalNAc-III-Tris-3 Step-1: Synthesis of GalNAc-III-Tris-2 To the solution of GalNAc-III-Tris-1 (0.055 g, 0.0205 mmol) in DMF were added NHS ester (0.090 g, 0.206 mmol), and DIPEA (0.035 mL, 0.206 mmol), and the resulting solution was allowed to stir for 1 hour. The reaction completion was observed by LCMS, and the product was purified by HPLC using 5-55 acetonitrile. LCMS m/z calc’d for C 143 H 202 N 21 O 48 S (M + H) + 3013.37; found 3013.67.
  • Step-2 Synthesis of GalNAc-III-Tris-3
  • DCM DCM
  • trifluoracetic acid: triisopropylsilane 95:5, 0.5 mL
  • the volatiles in the reaction were concentrated under rotavapor.
  • the crude product was precipitated in cold diethyl ether solution and centrifuged at 2,000 rpm for 10 mins. The supernatant was removed to get GalNAc-III-Tris-3 as a colorless solid.
  • Step 2 To a solution of 2-(pyridine-2-yl-disulfanyl) ethanol (2.0, 8.94 mmol), and TEA (1.25 mL, 8.94 mmol) in dry DCM (35 mL) at -780 °C, triphosgene (1.74 g, 5.98 mmol) in DCM (15 mL) was added dropwise and the reaction mixture was stirred at the same temperature for 2 h. Then, a solution of HOBT (1.58 g, 11.71 mmol) in DCM (15 mL) was added dropwise.
  • reaction mixture was slowly transferred to room temperature and stirred for 16 h. Then, the reaction mixture was diluted with DCM and washed twice with DI water followed by brine. Then, the organic solvent was dried over Na2SO4, and the solvent was evaporated under vacuum.
  • the crude compound was purified using flash column chromatography (silica gel, Hexane/EtOAc, 0-100% over 50 mins) to yield the pure magic carbonate product as a pale white solid (3.49 g, 96%).
  • Step 3 To a solution of magic carbonate (500 mg, 1.44 mmol) and DIPEA (1 mL, 5.76 mmol) in dry DMSO (5 mL) at 0 °C, triiodothyronine (938 mg, 1.44 mmol) was added. The reaction mixture was allowed to stir at 0 °C for 15 min and 1 h r. t. The completion of the reaction was confirmed by LCMS. Then, the compound was purified using reverse-phase flash column chromatography using a C18 column to obtain the T3-MC compound as a white solid (809 mg, 65%). LCMS m/z calc’d for C 23 H 19 I 3 N 2 O 6 S 2 (M + H) + 864.79; found 864.7.
  • Example 10 Synthesis of GalNAc-III-T3
  • 1,2-diaminoethane trityl resin 0.8-1.2 meq/g, 200-400 mesh
  • DCM dichloromethane
  • DMF dimethylformamide
  • GalNAc-IV-Tris-1 was cleaved from the resin using a cocktail solution of DCM: TFA: triisopropylsilane (TIPS) (55:40:5) (3 ⁇ 20 mL) for 20 minutes each time. The compound was concentrated under vacuum. Excess TFA was evaporated under rotavapor.
  • GalNAc-IV-NIR will be synthesized via the reaction between GalNAc-IV-Tris 1 compound and NIR-acid compound, followed by the O-deacetylation reaction.
  • Example 13 Step-1 Synthesis of GalNAc conjugate IV-Tris-2
  • a solution of GalNAc-IV-Tris-1 0.055 g, 0.0205 mmol
  • propanoic acid 3-[(triphenylmethyl)thio]-, 2,5-dioxo-1-pyrrolidinyl ester (NHS ester) (0.090 g, 0.206 mmol) and DIPEA (0.035 mL, 0.206 mmol) were added, and the resulting solution was stirred for 1 hour.
  • the reaction completion was confirmed by LCMS, and the product was purified by HPLC using a gradient of 5-55% acetonitrile.
  • mice were acclimatized for a week before starting each experiment. GalNAc-III-NIR or competitive ligand conjugate alone was administrated intravenously to mice once at 10 nmol. Mice were exposed to the near-infrared fluorescent light 4 hours after administration of the imaging agent. Mice were anesthetized before exposure to the near infrared red light at 725-810 nm for a maximum of 30 seconds. Mice were euthanized to assess the biodistribution of the imaging agent as shown in Fig.5B.
  • GalNAc-III-NIR and GalNAc-I-NIR were used to measure retention time. Each compound (1 nmol) was injected into two mice at different time points up to four days. The results are shown in Figs.6A-6G.
  • Example 15 Plasma Stability A plasma stability experiment was performed to determine the stability of GalNAc-III- T3 (GalNAc-III-64) in human plasma. GalNAc-III-T3 was tested in human plasma over a period of 8 hours (Fig.10A) and 24 hours (Fig.10B). The half-life (t 1/2 ) of GalNAc-III-T3 in human plasma was determined to be 60 minutes.
  • Example 16 Receptor Saturation A saturation binding experiment was performed for two mice receiving a concentration of 0.2, 0.4, 0.8, 1.6, 3.2, 6.4, or 12 nmol of GalNAc-III-NIR (GalNAc conjugate III-56 containing S0456). The maximum tolerable dose was 10 nmol. The results are shown in Fig. 8.
  • HFD high fat diet
  • mice received a regular diet (Mucedola srl) all throughout the experimental time. All animals were sacrificed under isoflurane anesthesia 21 weeks after the beginning of HFD or regular diet administration. Blood and tissues, including liver, intestine, heart, and kidney, were collected. Before sacrifice, blood samples were collected from a heart puncture, placed for 30 min on ice, and then centrifuged (10,000 g, 10 min, 4°C). The serum was collected for triglycerides (TG), total cholesterol (TC), aspartate aminotransferase (AST), and alanine transaminase (ALT) tests. All assays were performed following the manufacturer’s instructions using Cyman colorimetric activity assay kits.
  • liver samples were weighted, and sections were fixed in 10% buffered formalin and processed for histological analysis (H&E) or Oil O Red. The remaining tissues were snap-frozen in melting isobutyl alcohol or in liquid nitrogen and stored at ⁇ 80 ⁇ C until use.
  • H&E histological analysis
  • Oil O Red Oil Red O
  • isobutyl alcohol-frozen liver sections were stained with Oil Red O (ORO, Sigma Aldrich, St. Louis, MO, USA) for 15 min, rinsed with 60% isopropanol, and stained with Mayer hematoxylin (Sigma Aldrich). The ORO staining positive area for each sample was quantified by using ImageJ analysis software.
  • a range format should be interpreted in a flexible manner to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range were explicitly recited.
  • a range of “about 0.1% to about 5%” or “about 0.1% to 5%” should be interpreted to include not just about 0.1% to about 5%, but also the individual values (e.g., 1%, 2%, 3%, and 4%) and the sub-ranges (e.g., 0.1% to 0.5%, 1.1% to 2.2%, 3.3% to 4.4%) within the indicated range.

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Abstract

Conjugués qui ciblent un récepteur d'asialoglycoprotéine, p. ex. dans le foie, et comprennent un agent actif (p. ex. un agent thérapeutique ou d'imagerie) et des composants de liaison rigides ; compositions pharmaceutiques ; et procédés d'utilisation dans l'administration de conjugués, ainsi que l'imagerie et le traitement du foie (p. ex. chez un sujet atteint de NASH) à l'aide de conjugués.
PCT/US2025/020963 2024-03-22 2025-03-21 Ligands ciblant le récepteur d'asialoglycoprotéine spécifique du foie, conjugués les comprenant, compositions et procédés d'utilisation associés Pending WO2025199466A1 (fr)

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Citations (21)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018223073A1 (fr) 2017-06-02 2018-12-06 Wave Life Sciences Ltd. Compositions d'oligonucléotides et leurs procédés d'utilisation
WO2018223081A1 (fr) 2017-06-02 2018-12-06 Wave Life Sciences Ltd. Compositions d'oligonucléotides et leurs procédés d'utilisation
US20190111073A1 (en) 2017-10-16 2019-04-18 Roche Innovation Center Copenhagen A/S NUCLEIC ACID MOLECULE FOR REDUCTION OF PAPD5 AND PAPD7 mRNA FOR TREATING HEPATITIS B INFECTION
US20190216846A1 (en) 2016-06-17 2019-07-18 Hoffmann-La Roche Inc. Nucleic acid molecules for reduction of papd5 or papd7 mrna for treating hepatitis b infection
US20190343864A1 (en) 2016-12-13 2019-11-14 Am Sciences Inc Pharmaceutical composition for preventing or treating hepatitis b
US20200171069A1 (en) 2017-10-20 2020-06-04 Dicerna Pharmaceuticals, Inc. Methods for treating hepatitis b infection
US20200190523A1 (en) 2017-08-14 2020-06-18 Scandiedge Therapeutics Ab Pklr inhibition in the treatment of nafld and hcc
US20210024934A1 (en) 2018-04-05 2021-01-28 Hoffmann-La Roche, Inc. Use of fubp1 inhibitors for treating hepatitis b virus infection
US20210395745A1 (en) 2019-02-07 2021-12-23 Arrowhead Pharmaceuticals, Inc. Rnai agents for hepatitis b virus infection
US20210403908A1 (en) 2020-06-22 2021-12-30 Janssen Pharmaceuticals, Inc. Compositions and methods for treatment of hepatitis d virus infection
US20220378920A1 (en) 2019-09-10 2022-12-01 Daiichi Sankyo Company, Limited CONJUGATE OF GalNAc-OLIGONUCLEOTIDE FOR DELIVERY TO LIVER AND MANUFACTURING METHOD THEREOF
US20230097256A1 (en) 2020-01-31 2023-03-30 Avilar Therapeutics, Inc. Asgpr-binding compounds for the degradation of extracellular proteins
US20230118138A1 (en) 2019-12-19 2023-04-20 Hoffmann-La Roche Inc. Use of scamp3 inhibitors for treating hepatitis b virus infection
US20230122751A1 (en) 2019-12-19 2023-04-20 Hoffmann-La Roche Inc. Use of cops3 inhibitors for treating hepatitis b virus infection
US20230120063A1 (en) 2019-12-19 2023-04-20 Hoffmann-La Roche Inc. Use of saraf inhibitors for treating hepatitis b virus infection
US20230119360A1 (en) 2019-12-24 2023-04-20 Hoffmann-La Roche Inc. Pharmaceutical combination of a therapeutic oligonucleotide targeting hbv and a tlr7 agonist for treatment of hbv
US20230183692A1 (en) 2019-12-19 2023-06-15 Hoffmann-La Roche Inc. Use of sept9 inhibitors for treating hepatitis b virus infection
US20230193263A1 (en) 2019-12-19 2023-06-22 Hoffmann-La Roche Inc. Use of sbds inhibitors for treating hepatitis b virus infection
US20230257748A1 (en) 2020-08-21 2023-08-17 Hoffmann-La Roche Inc. Use of a1cf inhibitors for treating hepatitis b virus infection
US20230295630A1 (en) 2022-03-16 2023-09-21 Empirico Inc. Galnac compositions for improving sirna bioavailability
US20230357773A1 (en) 2020-08-05 2023-11-09 Hoffmann-La Roche Inc. Oligonucleotide treatment of hepatitis b patients

Patent Citations (22)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20190216846A1 (en) 2016-06-17 2019-07-18 Hoffmann-La Roche Inc. Nucleic acid molecules for reduction of papd5 or papd7 mrna for treating hepatitis b infection
US20190343864A1 (en) 2016-12-13 2019-11-14 Am Sciences Inc Pharmaceutical composition for preventing or treating hepatitis b
WO2018223073A1 (fr) 2017-06-02 2018-12-06 Wave Life Sciences Ltd. Compositions d'oligonucléotides et leurs procédés d'utilisation
WO2018223081A1 (fr) 2017-06-02 2018-12-06 Wave Life Sciences Ltd. Compositions d'oligonucléotides et leurs procédés d'utilisation
US20200190523A1 (en) 2017-08-14 2020-06-18 Scandiedge Therapeutics Ab Pklr inhibition in the treatment of nafld and hcc
US20190111073A1 (en) 2017-10-16 2019-04-18 Roche Innovation Center Copenhagen A/S NUCLEIC ACID MOLECULE FOR REDUCTION OF PAPD5 AND PAPD7 mRNA FOR TREATING HEPATITIS B INFECTION
US20200171069A1 (en) 2017-10-20 2020-06-04 Dicerna Pharmaceuticals, Inc. Methods for treating hepatitis b infection
US20210024934A1 (en) 2018-04-05 2021-01-28 Hoffmann-La Roche, Inc. Use of fubp1 inhibitors for treating hepatitis b virus infection
US20210395745A1 (en) 2019-02-07 2021-12-23 Arrowhead Pharmaceuticals, Inc. Rnai agents for hepatitis b virus infection
US20220378920A1 (en) 2019-09-10 2022-12-01 Daiichi Sankyo Company, Limited CONJUGATE OF GalNAc-OLIGONUCLEOTIDE FOR DELIVERY TO LIVER AND MANUFACTURING METHOD THEREOF
US20230118138A1 (en) 2019-12-19 2023-04-20 Hoffmann-La Roche Inc. Use of scamp3 inhibitors for treating hepatitis b virus infection
US20230122751A1 (en) 2019-12-19 2023-04-20 Hoffmann-La Roche Inc. Use of cops3 inhibitors for treating hepatitis b virus infection
US20230120063A1 (en) 2019-12-19 2023-04-20 Hoffmann-La Roche Inc. Use of saraf inhibitors for treating hepatitis b virus infection
US20230183692A1 (en) 2019-12-19 2023-06-15 Hoffmann-La Roche Inc. Use of sept9 inhibitors for treating hepatitis b virus infection
US20230193263A1 (en) 2019-12-19 2023-06-22 Hoffmann-La Roche Inc. Use of sbds inhibitors for treating hepatitis b virus infection
US20230119360A1 (en) 2019-12-24 2023-04-20 Hoffmann-La Roche Inc. Pharmaceutical combination of a therapeutic oligonucleotide targeting hbv and a tlr7 agonist for treatment of hbv
US20230097256A1 (en) 2020-01-31 2023-03-30 Avilar Therapeutics, Inc. Asgpr-binding compounds for the degradation of extracellular proteins
US20240072809A1 (en) 2020-01-31 2024-02-29 Avilar Therapeutics, Inc. Asgpr-binding compounds for the degradation of extracellular proteins
US20210403908A1 (en) 2020-06-22 2021-12-30 Janssen Pharmaceuticals, Inc. Compositions and methods for treatment of hepatitis d virus infection
US20230357773A1 (en) 2020-08-05 2023-11-09 Hoffmann-La Roche Inc. Oligonucleotide treatment of hepatitis b patients
US20230257748A1 (en) 2020-08-21 2023-08-17 Hoffmann-La Roche Inc. Use of a1cf inhibitors for treating hepatitis b virus infection
US20230295630A1 (en) 2022-03-16 2023-09-21 Empirico Inc. Galnac compositions for improving sirna bioavailability

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
"Remington: The Science & Practice of Pharmacy", 2005, LIPPINCOTT WILLIAMS & WILKINS
KHATRI VINOD ET AL: "Design, Synthesis, and Biomedical Applications of Glycotripods for Targeting Trimeric Lectins", EUROPEAN JOURNAL OF ORGANIC CHEMISTRY, vol. 26, no. 9, 1 March 2023 (2023-03-01), DE, XP093051398, ISSN: 1434-193X, Retrieved from the Internet <URL:https://onlinelibrary.wiley.com/doi/full-xml/10.1002/ejoc.202201360> DOI: 10.1002/ejoc.202201360 *
MARTIN: "Remington's Pharmaceutical Sciences", 1990, MACK PUBLISHING COMPANY
SCHMIDT ET AL., NUCL ACIDS RES, vol. 45, 2017, pages 2294

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