ASGPR cell surface receptor binding compounds and conjugates

Abstract

The present disclosure provides certain compounds that include a ligand moiety that specifically binds to a cell surface asialoglycoprotein receptor (ASGPR). The cell surface ASGPR binding compound can trigger the receptor to internalize the bound compound into the cell. The ligand moiety of the present disclosure can be linked to various moieties of interest without affecting the specific binding to the cell surface receptor ASGPR and the function of the cell surface receptor ASGPR. Also provided are compounds that are conjugates of a ligand moiety linked to a biomolecule, such as an antibody, that can utilize a cellular pathway to remove a particular protein of interest from the cell surface or the extracellular environment. Also provided herein are methods of using the conjugates to sequester and / or target a polypeptide of interest for lysosomal degradation.

Description

[Technical Field]

[0001] 1. CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. 63 / 221,918, filed July 14, 2021, which is hereby incorporated by reference in its entirety. [Background technology]

[0002] 2. Introduction Many therapeutic agents act by binding to functionally important sites in target proteins, thereby modulating the activity of those proteins, or by recruiting immune effectors, such as many monoclonal antibody drugs, to act on target proteins. However, there exists an untapped treasure trove of medically important human proteins that are considered "undruggable" because these proteins are not readily amenable to currently available therapeutic targeting approaches. Therefore, there is a need for therapeutic agents that can target a wider range of proteins.

[0003] The asialoglycoprotein receptor (ASGPR), also known as the Ashwell-Morrell receptor, is a transmembrane glycoprotein receptor present primarily in hepatocytes that plays a key role in serum glycoprotein homeostasis by mediating the endocytosis and lysosomal degradation of glycoproteins with exposed terminal galactose or N-acetylgalactosamine (GalNAc) residues. The ASGPR cycles between endosomes and the cell surface.

[0004] Alternative ligands that bind to cell surface ASGPRs and subsequently cause transport across the cell membrane are of great interest. Summary of the Invention

[0005] 3. Summary of the Invention The present disclosure provides certain compounds comprising a ligand moiety that specifically binds to a cell surface asialoglycoprotein receptor (ASGPR). Compounds that bind to the cell surface ASGPR can trigger the receptor to internalize the bound compound into the cell. The ligand moieties of the present disclosure can be linked to various moieties of interest without affecting their specific binding to the cell surface receptor ASGPR and its function. Also provided are compounds that are conjugates of the ligand moiety linked to a biomolecule, such as an antibody, which can utilize a cellular pathway to remove a specific protein of interest from the cell surface or extracellular environment. For example, the conjugates described herein may sequester and / or degrade a target molecule of interest in the lysosomes of a cell. Also provided herein are compositions comprising such conjugates, as well as methods of using the conjugates to sequester and / or target a polypeptide of interest for lysosomal degradation, and methods of using the conjugates to treat a disorder or disease. 4. Brief description of the drawings These and other features, aspects, and advantages of the present invention will become better understood with reference to the following description and accompanying drawings, in which: [Brief explanation of the drawings]

[0006] [Figure 1] FIG. 1 is a graph of cellular fluorescence (MFI) versus antibody conjugate concentration ([Ab]) showing that various antibody conjugates of an exemplary ASGPR-binding compound and an exemplary M6PR-binding compound (520) exhibited comparable potent uptake into HepG2 cells after 1 hour of incubation. [Figure 2A] 2A-2D are graphs of cellular fluorescence versus antibody conjugate concentration showing that various antibody conjugates of exemplary ASGPR-binding compounds exhibited strong uptake into HepG2 cells after 1 hour of incubation. [Figure 2B]2A-2D are graphs of cellular fluorescence versus antibody conjugate concentration showing that various antibody conjugates of exemplary ASGPR-binding compounds exhibited strong uptake into HepG2 cells after 1 hour of incubation. [Figure 2C] 2A-2D are graphs of cellular fluorescence versus antibody conjugate concentration showing that various antibody conjugates of exemplary ASGPR-binding compounds exhibited strong uptake into HepG2 cells after 1 hour of incubation. [Figure 2D] 2A-2D are graphs of cellular fluorescence versus antibody conjugate concentration showing that various antibody conjugates of exemplary ASGPR-binding compounds exhibited strong uptake into HepG2 cells after 1 hour of incubation.

[0007] [Figure 3] FIG. 1 shows the results of screening exemplary trivalent compounds (1901 (I-171), 1902 (I-172), XB32, and 2101) by fluorescence polarization.

[0008] [Figure 4] FIG. 1 shows the binding of example monovalent compounds (591, XB20, XB23, XB21, 592, and 593) as a percentage of the activity of the reference compound 18G.

[0009] [Figure 5] FIG. 1 shows the results of screening monovalent compounds (XB20, XB21, 592, XB23, 591) by fluorescence polarization as examples.

[0010] [Figure 6] 1 is a graph of the cellular uptake in HepG2 cells of various conjugates of OMA (anti-IgE) with example compounds I-160 to I-163 and I-141 bound to Alexa488-labeled target IgE.

[0011] [Figure 7] FIG. 1 shows the affinity-dependent clearance of OMA-example compounds (I-160 to I-163) compared to OMA (reference).

[0012] [Figure 8] FIG. 1 shows the dose escalation method for OMA-I-163 IgE clearance.

[0013] [Figure 9] FIG. 1 shows affinity-dependent clearance of OMA-example compounds (I-160 to I-163) compared to hIgE (reference). DETAILED DESCRIPTION OF THE INVENTION

[0014] 5. MODE FOR CARRYING OUT THE INVENTION As summarized above, the present disclosure provides various compounds comprising a ligand moiety that specifically binds to a cell surface receptor. Also provided herein are conjugates comprising a moiety X that binds to such a cell surface receptor, e.g., an internalized cell surface receptor for sequestration and / or lysosomal degradation. In certain embodiments, the cell surface receptor is ASGPR.

[0015] The present disclosure provides a compound of formula (I): [ka] (In the formula, X is a moiety (e.g., as described herein) that binds to an ASGPR cell surface receptor; n is 1 to 500; L is a linker of defined length (e.g., monovalent or multivalent, as described herein); Y is a moiety of interest (e.g., as described herein) or a salt thereof.

[0016] The compounds and conjugates, and methods of the present disclosure are described in further detail below. Certain ASGPR-binding compounds are described. Biomolecular conjugates comprising a cell surface receptor-binding moiety (X) that binds to ASGPR are also described. Linkers (L) and moieties of interest (Y) used in the ASGPR-binding compounds and biomolecular conjugates are also described. Methods in which the compounds and conjugates of the present disclosure can be used are also described.

[0017] 5.1.ASGPR Ligands As summarized above, the present disclosure provides certain compounds comprising a ligand moiety that specifically binds to cell surface ASGPR. The ASGPR ligand moiety of the present disclosure can be linked to various moieties of interest without affecting the specific binding to and function of the cell surface ASGPR. The inventors have demonstrated that the compounds of the present disclosure can exploit the function of cell surface ASGPR in biological systems, for example, for the internalization and sequestration of the compound into lysosomes of cells, and in some cases subsequent lysosomal degradation. The compounds of the present disclosure are used in a variety of applications.

[0018] The term "asialoglycoprotein receptor" (ASGPR) (also known as Ashwell-Morrell receptor) refers to a transmembrane glycoprotein receptor present primarily in hepatocytes that plays a key role in serum glycoprotein homeostasis by mediating the endocytosis and lysosomal degradation of glycoproteins with exposed terminal galactose or N-acetylgalactosamine (GalNAc) residues. ASGPR cycles between endosomes and the cell surface. In certain embodiments, the ASGPR is Homo sapiens asialoglycoprotein receptor 1 (ASGR1) (see, e.g., NCBI Reference Sequence: NM_001197216).

[0019] Compounds comprising such an ASGPR-binding moiety (X) (e.g., as described herein) may bind to other receptors, and may bind with lower affinity, e.g., as measured by immunoassays or other assays known in the art. In certain embodiments, X, or a compound described herein comprising such X, specifically binds to the cell surface ASGPR with an affinity that is at least 2 logs, 2.5 logs, 3 logs, 4 logs, or more than the affinity with which X, or the compound or conjugate, binds to another cell surface receptor. In certain embodiments, X, or a compound described herein comprising X, specifically binds to the cell surface ASGPR with an affinity (K d ) specifically binds to ASGPR. In certain embodiments, such binding affinity (K d ) is 10 mM or less, 1 mM or less, 100 uM or less, 10 uM or less, 1 uM or less, 100 nM or less, 10 nM or less, or 1 nM or less. In this context, the terms "bind," "bind to," "specifically bind," or "specifically bind to" are used interchangeably.

[0020] The ASGPR-binding compounds of the present disclosure comprise a moiety (X) that specifically binds to the cell surface receptor ASGPR. The ASGPR-binding compounds may be monovalent or multivalent (e.g., bivalent or trivalent or higher), where a monovalent compound contains a single ASGPR ligand moiety and a multivalent compound contains two or more such moieties.

[0021] In certain embodiments, the ASGPR-binding moiety X is capable of binding to a specific cell-surface ASGPR and directing (or targeting) the molecule to this receptor. In certain embodiments, the ASGPR-binding moiety X is capable of binding to an ASGPR and directing (or targeting) a compound or conjugate described herein to lysosomes for internalization and sequestration and / or subsequent lysosomal degradation.

[0022] In some embodiments, the ASGPR-binding moiety X comprises an amino sugar ring derivative of galactose (e.g., N-acetylgalactosamine and its analogs), which is linked to the 1-, 6-, or 2-position of the sugar ring via a linking moiety. The length of the linking moiety can be 1 to 10 atoms, e.g., 1 to 6 atoms, or 1 to 5 atoms, or 1 to 4 atoms, or 1 to 3 atoms. In some embodiments, the amino sugar ring derivative of galactose is linked to the oxygen, sulfur, nitrogen, or carbon atom at the 1-position of the ring via a linking moiety. In some embodiments, the amino sugar ring derivative of galactose is linked to the oxygen, sulfur, nitrogen, or carbon atom at the 6-position of the ring via a linking moiety. In some embodiments, the amino sugar ring derivative of galactose is linked to the oxygen, sulfur, nitrogen, or carbon atom at the 2-position of the ring via a linking moiety. In certain embodiments, the amino sugar ring derivative of galactose is linked to the heteroaryl group at the 1-, 6-, or 2-position of the ring via a linking moiety. In certain embodiments, the amino ring derivative of galactose is a bicyclic structure.

[0023] In some embodiments, the ASGPR-binding compound is monovalent (e.g., in Formula (I), n is 1), and thus the ASGPR-binding compound comprises a single ASGPR ligand moiety (X) linked to a moiety of interest (Y) at position 1, 6, or 2 of (X) via a linking moiety. In certain embodiments of Formula (I), n is 1, and L comprises a linear linker having a backbone of 20 or more contiguous atoms covalently linking the ASGPR ligand (X) to Y via a linking moiety at either position 1, 2, or 6 of X. In some cases, n is 20 to 100, e.g., 25 to 80, 25 to 60, or 25 to 50 contiguous atoms. In certain embodiments of Formula (I), n is 1, and L comprises a backbone of 25 or more contiguous atoms covalently linking the ASGPR ligand (X) to Y.

[0024] In some embodiments, the ASGPR-binding compound is multivalent (e.g., in Formula (I), n is 2 or greater, such that the ASGPR-binding compound comprises a plurality of ASGPR ligand-binding moieties (X), each covalently linked to a moiety of interest (Y) via a branched linker (e.g., L is a branched linker). In certain cases, the ASGPR-binding compound is bivalent (e.g., in Formula I, n is 2). In other certain cases, the ASGPR-binding compound is trivalent (e.g., in Formula I, n is 3). In certain cases, each branch of the branched linker is linked to the linking moiety of each X. to a branch point of the linker. In certain cases, each branch of the linker comprises 14 to 50 consecutive atoms, e.g., 14 to 40, 14 to 30, or 14 to 20 atoms, connecting the two. In certain cases, each branch of the linker comprises a linear linker of 20 or more consecutive atoms. In certain cases, the linker comprises a linear linker of 12 or more consecutive atoms, e.g., 15 or more, 20 or more, 30 or more, or even more consecutive atoms, covalently connecting the branch point of L to the moiety of interest (Y).

[0025] The ASGPR binding moiety (X) of the compounds of the present disclosure has formula (Ia): [ka] (In the formula, R 1 -OH, -OC(O)R, -C(O)NHR, -Z 1 -*, and optionally substituted triazole, provided that R is optionally substituted C 1~6 alkyl or optionally substituted aryl; R 2 is -NHCOCH3, -NHCOCF3, -NHCOCH2CF3, -OH, optionally substituted triazole, and -Z 1 - Selected from *, R3 -H, -OH, -CH3, -OCH3, -OCH2CH=CH, and -Z 1 - Selected from *, R 1 ~R 3 One of them is -Z 1 -* or -Z 1 - *, where * is Z to the linker (L) 1 represents the point of attachment of R 4 and R 5 are each independently selected from H and a promoiety; or R 4 and R 5 are linked in a ring to form a pro-moiety, R 11 is H or a group that forms a bridge to the carbon atom at position 1 (e.g., a bridge of two atoms linked in a ring), Z 1 is Z 11 , optionally replaced by Z 11 -heteroaryl, optionally substituted Z 11 - a linking moiety selected from aryl, optionally substituted heteroaryl, optionally substituted aryl, optionally substituted alkyl, optionally substituted amido, optionally substituted sulfonamido, optionally substituted urea, and optionally substituted thiourea; Z 11 -O-, -S-, NR 21 - and -C(R 22 )2 is selected, Each R 21 are independently selected from H and optionally substituted (C1-C6) alkyl; Each R 22 are independently selected from H, halogen (e.g., F), and optionally substituted (C1-C6) alkyl. It can be expressed as:

[0026] In some embodiments of Formula (Ia), i) n is 3 and R 1is OH and R 2 is -NHCOCH3 and R 4 and R 5 is H and R 3 Z 1 If Z 1 is not O, ii) n is 2 or 3, and R 1 is OAc and R 2 is -NHCOCH3 and R 4 and R 5 is Ac and R 3 Z 1 If Z 1 is not O, iii) n is 2 or 3, and R 1 is Obz and R 2 is -NHCOCH3 and R 4 and R 5 is Bz and R 3 Z 1 If Z 1 is not O, iv) n is 3 and R 1 is OH and R 2 is -NHCOCH3 and R 4 and R 5 is H and R 3 Z 1 and Z 11 is O, then L comprises a main chain of at least 16 contiguous atoms up to a branch point; v) n is 3 and R 1 Z 1 However, Z 1 is O and R 4 and R 5 If is H, then R 3 is not -CH3-OCH3 or -OCH2CH=CH, vi)R 11 is a group of formula -CHO- that forms a bridge (i.e., cyclic link) to the carbon atom at position 1 of the sugar ring, and R 2 is -NHCOCH3 and R 4 and R 5 If is H, then R 1 and R3 is Z 1 isn't it.

[0027] In some embodiments, the ASGPR binding moiety (X) of a compound of the present disclosure has formula (Ia-1): [ka] (In the formula, R 1 -OH, -OC(O)R, -C(O)NHR, -Z 1 -*, and optionally substituted triazole, provided that R is optionally substituted C 1~6 alkyl or optionally substituted aryl; R 2 is -NHCOCH3, -NHCOCF3, -NHCOCH2CF3, -OH, optionally substituted triazole, and -Z 1 - Selected from *, R 3 -H, -OH, -CH3, -OCH3, -OCH2CH=CH, and -Z 1 - Selected from *, R 1 ~R 3 One of them is -Z 1 -*, where "*" is Z for the linker (L). 1 represents the point of attachment of R 4 and R 5 are each independently selected from H and a promoiety (e.g., an ester promoiety); Z 1 is Z 11 , optionally replaced by Z 11 -heteroaryl, optionally substituted Z 11 - a linking moiety selected from aryl, optionally substituted heteroaryl, optionally substituted aryl, optionally substituted amide, optionally substituted sulfonamide, optionally substituted urea, and optionally substituted thiourea; Z 11 -O-, -S-, NR 21 - and -C(R 22)2 is selected, Each R 21 are independently selected from H and optionally substituted (C1-C6) alkyl; Each R 22 are independently selected from H, halogen (e.g., F), and optionally substituted (C1-C6) alkyl. It can be expressed as:

[0028] In some embodiments of Formula (Ia-1), i) n is 3 and R 1 is OH and R 2 is -NHCOCH3 and R 4 and R 5 is H and R 3 Z 1 If Z 1 is not O, ii) n is 2 or 3 and R 1 is OAc and R 2 is -NHCOCH3 and R 4 and R 5 is Ac and R 3 Z 1 If Z 1 is not O, iii) n is 2 or 3, and R 1 is OBz and R 2 is -NHCOCH3 and R 4 and R 5 is Bz and R 3 Z 1 If Z 1 is not O, iv) n is 3 and R 1 is OH and R 2 is -NHCOCH3 and R 4 and R 5 is H and R 3 Z 1 and Z 1 is O, then L comprises a main chain of at least 16 consecutive atoms up to the branch point; and / or v) n is 3 and R 1 Z1 However, Z 1 is O and R 4 and R 5 If is H, then R 3 is not -CH3.

[0029] In some embodiments, the ASGPR binding moiety (X) of a compound of the present disclosure has formula (Ib): [ka] (In the formula, R 1 , R 2 , R 4 , R 5 , and Z 1 is as defined herein) It can be expressed as:

[0030] In some embodiments of Formula (Ib), Z1 is in the alpha configuration: [ka] is located.

[0031] In some embodiments of Formula (Ib), Z 1 Beta deployment: [ka] is located.

[0032] In certain embodiments of Formula (Ib), Z 1 is Z 11 -Ar, where Ar is an optionally substituted heteroaryl or an optionally substituted aryl. Optionally, Ar is an optionally substituted heteroaryl. Optionally, the heteroaryl is a 5- or 6-membered heteroaryl. Optionally, the heteroaryl is a 5-membered heteroaryl. Optionally, the 5-membered heteroaryl is a triazole. Optionally, the triazole is a 1,2,3-triazole moiety. Optionally, Z 11 -C(R 22)2. In some cases, at least one of R 22 is H. In some cases, both R 22 is H. In some cases, Z 11 is -O-. In some cases, Z 11 is -S-. In certain other cases, Z 11 Ha-NR 21 where R 21 is H or (C 1~3 ) alkyl. 1 -C(R 22 ) 2-triazole-. Optionally, Z 1 teeth [ka] In some cases, Z 1 teeth [ka] is.

[0033] In certain embodiments of Formula (Ib), Z 1 is Z 11 In some cases, Z 11 -C(R 22 )2. In some cases, at least one of R 22 is H. In some cases, both R 22 is H and Z 11 is -CH2-. In some cases, Z 11 is -O-. In some cases, Z 11 is -S-. In certain other cases, Z 11 Ha-NR 21 where R 21 is H or (C1-3) alkyl.

[0034] In certain embodiments of Formula (Ib), Z 1 is a monocyclic 5- or 6-membered heteroaryl or aryl. 1 teeth [ka] In some cases, Z 1 teeth [ka] is.

[0035] In certain embodiments of Formula (Ib), Z 1 -O-, -S-, -C(R 22 )2-, -NR 21 -,-CONR 21 - and [ka] (In the formula, X 1 is O or S, t is 0 or 1, R 21 and each R 23 are independently selected from H and optionally substituted (C1-C6) alkyl (e.g., C such as methyl). (1~3) alkyl), Each R 22 are independently selected from H, halogen (e.g., F), and optionally substituted (C1-C6) alkyl. is selected from.

[0036] In certain embodiments of Formula (Ib), Z 1 is an optionally substituted (C1-C6) alkyl. Z 1 In some cases, the alkyl is methyl. 1 In some cases, the alkyl is ethyl. 1 In some cases, the alkyl is propyl. 1 In some cases, the alkyl is butyl. 1 In some cases, the alkyl is pentyl. 1 In some cases, the alkyl is hexyl.

[0037] In certain embodiments, the compound of formula (Ib) above has the following structure: [ka] (In the formula, R 5 are independently H or pro moieties) is selected from one of the following:

[0038] In some embodiments, the compound of formula (Ib) above has the following structure: [ka] (In the formula, R 5 and R 4 are independently H or a pro-moiety, or R 4 and R 5 are linked in a ring to form a pro-moiety, n1 is an integer from 1 to 6. is selected from one of the following:

[0039] In some embodiments, R 4 and R 5 At least one of R is of formula -COCH3, -COCH(CH3)2, or -COC(CH3)3. 4 and R 5 At least one of R is of formula -CHOCOC(CH). 4 and R 5 At least one of R is of formula -COC(CH3)3 or -CH2OCOC(CH3)3. 4 is H and R 5 is selected from -COCH, -COCH(CH), -COC(CH), and -CHOCOC(CH). 4 is H and R 5 is -COC(CH3)3. In some cases, R 4 is H and R 5 is —CH 2 OCOC(CH 3 ) 3. In some embodiments, the compound of formula (Ib) above has the following structure: [ka] (wherein n2 is an integer of 1 to 6) is selected from one of the following:

[0040] In some embodiments of compounds of Formula (Ib), R 5 and R 4 are linked in a ring to form a promoiety. Optionally, the compound of formula (Ib) has the following structure: [ka] (wherein n2 is an integer of 1 to 6, and Y 4 is an appropriate counterion) is selected from one of the following:

[0041] In some embodiments, the ASGPR binding moiety (X) of a compound of the present disclosure has formula (Ic): [ka] (In the formula, R 2 ~R 5 and Z 1 is as defined herein) It can be expressed as:

[0042] In certain embodiments of Formula (Ic), Z 1 -O-, -S-, -CONR 21 - and optionally substituted -(C(R 22 )2) q -heteroaryl, where q is 0 or 1. 1 is -O-. In certain other cases, Z 1 is optionally substituted -(C(R 22 )2) q -triazole, where q is 0 or 1. 1 teeth [ka] is.

[0043] In certain embodiments of Formula (Ic), Z 1 is Z 11 -Ar, where Ar is an optionally substituted heteroaryl or an optionally substituted aryl. Optionally, Ar is an optionally substituted heteroaryl. Optionally, said heteroaryl is a 5- or 6-membered heteroaryl. Optionally, said heteroaryl is a 5-membered heteroaryl. Optionally, said 5-membered heteroaryl is a triazole. Optionally, said triazole is a 1,2,3-triazole moiety. Optionally, Z 11 Ha-(C(R 22 )2. Optionally, at least one R 22 is H. In some cases, both R 22 is H. In some cases, Z 11 is -O-. Optionally, Z 11 is -S-. In some cases, Z 11 Ha-NR 21 (However, R 21 is H or (C1-3) alkyl). 1 -C(R 22 ) 2-triazole-. Optionally, Z 1 teeth [ka] is.

[0044] In certain embodiments of Formula (Ic), Z 1 is Z 11 In some cases, Z 11 -C(R 22 )2. In some cases, at least one of R 22 is H. In some cases, both R 22 is H and Z 11 is -CH2-. In some cases, Z 11 is -O-. In some cases, Z 11 is -S-. In certain other cases, Z 11 Ha-NR21 (However, R 21 is H or (C 1~3 ) alkyl).

[0045] In certain embodiments of Formula (Ic), Z 1 is a monocyclic 5- or 6-membered heteroaryl or aryl. 1 teeth [ka] is.

[0046] In certain embodiments of Formula (Ic), Z 1 -O-, -S-, -C(R 22 )2-, -NR 21 -,-CONR 21 - and [ka] (In the formula, X 1 is O or S, t is 0 or 1, R 21 and each R 23 are independently selected from H and optionally substituted (C1-C6) alkyl (e.g., C such as methyl). (1~3) alkyl), Each R 22 are independently selected from H, halogen (e.g., F), and optionally substituted (C1-C6) alkyl. is selected from.

[0047] In certain embodiments, the compound of formula (Ic) has the following structure: [ka] is.

[0048] In certain embodiments, the compound of formula (Ic) has the following structure: [ka] is.

[0049] In some embodiments, the ASGPR binding moiety (X) of a compound of the present disclosure has formula (Id): [ka] (In the formula, R 1 , R 3 ~R 5 and Z 1 is as defined herein) It can be expressed as:

[0050] In some embodiments, Z 1 is optionally substituted -(C(R 22 )2) q heteroaryl, and [ka] (wherein q is 0 or 1) is selected from.

[0051] In some embodiments, Z 1 is optionally substituted -(C(R 22 )2) q -triazole (wherein q is 0 or 1).

[0052] In some embodiments, Z 1 teeth [ka] In some embodiments, Z 1 teeth [ka] is.

[0053] In some embodiments, Z 1 teeth [ka] (In the formula, R 23 is H or C (1~3) -alkyl) is.

[0054] In some embodiments, Z 1 Ha-NR 23 CO-(However, R 23 is H or C (1~3) -alkyl).

[0055] In certain embodiments of Formula (Id), Z 1 is optionally substituted -(C(R 22 )2) q heteroaryl, and [ka] (wherein q is 0 or 1) is selected from.

[0056] In certain embodiments of Formula (Id), Z 1 is optionally substituted -(C(R 22 )2) q -triazole, where q is 0 or 1. 1 teeth [ka] is.

[0057] In some cases, Z 1 teeth [ka] (In the formula, R 23 is H or C (1~3) -alkyl) is.

[0058] In some cases, Z 1 Ha-NR 23 CO-(However, R 23 is H or C (1~3)-alkyl).

[0059] In certain embodiments of Formula (Id), Z 1 is a monocyclic 5- or 6-membered heteroaryl or aryl. 1 teeth [ka] is.

[0060] In certain embodiments of Formula (Id), Z 1 has the following structure: [ka] is a monocyclic 5- or 6-membered heteroaryl.

[0061] In certain embodiments of Formula (Id), Z 1 has the following structure: [ka] It is one of the following.

[0062] In certain embodiments of Formula (Id), Z 1 -O-, -S-, -C(R 22 )2-, -NR 21 -,-CONR 21 - and [ka] (In the formula, X 1 is O or S, t is 0 or 1, R 21 and each R 23 are independently selected from H and optionally substituted (C1-C6) alkyl (e.g., C such as methyl). (1~3) alkyl), Each R 22are independently selected from H, halogen (e.g., F), and optionally substituted (C1-C6) alkyl. is selected from.

[0063] In certain embodiments, the compound of formula (Id) has the following structure: [ka] (In the formula, R 6 are independently H or (C 1~3 ) alkyl) In some embodiments of the compound of Formula (Id), R 3 is H, so that compounds of formula (Id) have no non-hydrogen substituents at the 1-position of the sugar ring.

[0064] In some embodiments, the ASGPR binding moiety (X) of a compound of the present disclosure has formula (Ie′): [ka] (In the formula, R 1 , R 4 , R 5 , and R 11 is as defined herein; Z 2 is absent or -O-, -S-, NR 25 - and -C(R 22 )2 is selected, Ring A is absent or selected from 5- or 6-membered optionally substituted aryl and 5- or 6-membered optionally substituted heteroaryl; Z 3 is Z 12 , optionally substituted alkyl, optionally substituted Z 12 -alkyl, optionally substituted Z 12 -heteroaryl, optionally substituted Z 12- a linking moiety selected from aryl, optionally substituted heteroaryl, optionally substituted aryl, optionally substituted amide, optionally substituted sulfonamide, optionally substituted urea, and optionally substituted thiourea; Z 12 -CHO-, -O-, -S-, -NR 26 - and -C(R 22 )2- is selected from R 25 and R 26 are each independently H, optionally substituted (C1-C6) alkyl (e.g., C such as methyl), (1~3) -alkyl), and optionally substituted acyl; Each R 22 are independently selected from H, halogen (e.g., F), and optionally substituted (C1-C6) alkyl. It can be expressed as:

[0065] In some embodiments of Formula (Ie′), the ASGPR binding moiety (X) of the compounds of the present disclosure has the formula (Ie″): [ka] It can be expressed as:

[0066] In some embodiments of Formula (Ie′), the ASGPR binding moiety (X) of the compounds of the present disclosure has Formula (Ie): [ka] (In the formula, R 1 , R 4 , R 5 , and R 11 is as defined herein; Z 2 is absent or -O-, -S-, NR 25 - and -C(R 22 )2 is selected, Ring A is absent or selected from 5- or 6-membered optionally substituted aryl and 5- or 6-membered optionally substituted heteroaryl; Z 3 is Z 12 , optionally substituted alkyl, optionally substituted Z 12 -alkyl, optionally substituted Z 12 -heteroaryl, optionally substituted Z 12 - a linking moiety selected from aryl, optionally substituted heteroaryl, optionally substituted aryl, optionally substituted amide, optionally substituted sulfonamide, optionally substituted urea, and optionally substituted thiourea; Z 12 -CHO-, -O-, -S-, -NR 26 - and -C(R 22 )2- is selected from R 25 and R 26 are each independently H, optionally substituted (C1-C6) alkyl (e.g., C such as methyl), (1~3) -alkyl), and optionally substituted acyl; Each R 22 are independently selected from H, halogen (e.g., F), and optionally substituted (C1-C6) alkyl. It can be expressed as:

[0067] In some embodiments, Z 2 is absent. In some embodiments, Z 2 is C(R 22 )2(However, R 22 is H or optionally substituted (C1-3) alkyl. In some embodiments, Z 2 is -CH-. In some embodiments, Z 2 is NR 25 (However, R 25 is selected from H, optionally substituted (C1-3) alkyl, and optionally substituted acyl. In some embodiments, Z2 is —N(COCH)—. In some embodiments, Z 2 is -NH-. In some embodiments, Z 2 is -S-. In some embodiments, Z 2 is O.

[0068] In some embodiments, the compound of formula (Ie) has the formula (If)-(Ii): [ka] (In the formula, rings A, R 1 , R 4 , R 5 , R 11 , Z 3 , and R 25 is as defined herein) It is one of the following.

[0069] In some embodiments of any one of Formulas (Ie)-(Ii), Ring A is a 5- or 6-membered aryl or heteroaryl. Optionally, Ring A is a 5-membered heteroaryl selected from triazole, thiadiazole, thiophene, oxazole, isoxazole, isothiazole, thiazole, oxadiazole, and furan. Optionally, Ring A is a 6-membered heteroaryl selected from pyridine, pyrimidine, pyridazine, pyrazine, and triazine. Optionally, Ring A is triazole. Optionally, Ring A is pyridine. Optionally, Ring A is pyrimidine. Optionally, Ring A is thiadiazole.

[0070] In some embodiments of any one of Formulas (Ie)-(Ii), ring A is absent.

[0071] In some embodiments of any one of Formulas (Ie)-(Ii), ring A is phenyl or substituted phenyl.

[0072] In some embodiments of Formula (If), Ring A is a 5- or 6-membered heteroaryl. In certain cases of Formula (If), Ring A is a 5-membered heteroaryl. In certain embodiments of Formula (If), Ring A is triazole. In certain embodiments of (If), Ring A is absent.

[0073] In some embodiments of Formula (Ig), ring A is a 5- or 6-membered heteroaryl and R 25 is H. In certain embodiments of Formula (Ig), Ring A is triazole. In certain cases of Formula (Ig), Ring A is pyridine. In certain cases of Formula (Ig), Ring A is pyrimidine. In certain cases of Formula (Ig), Ring A is thiadiazole. In some embodiments of Formula (Ig), Ring A is absent and R 25 is H or optionally substituted acyl. 25 is -COCH3. In some cases, R 25 is H.

[0074] In some embodiments of Formula (Ih), ring A is a 5- or 6-membered heteroaryl. In certain cases of Formula (Ih), ring A is a 5-membered heteroaryl. In certain embodiments of Formula (Ih), ring A is triazole. In certain embodiments of (Ih), ring A is absent.

[0075] In some embodiments of Formula (Ii), Ring A is a 5- or 6-membered heteroaryl. In certain cases of Formula (Ii), Ring A is a 5-membered heteroaryl. In certain embodiments of Formula (Ii), Ring A is a triazole. In certain embodiments of (Ii), Ring A is absent.

[0076] In some cases, the ASGPR binding moiety (X) of the compounds of the present disclosure is represented by formula (Ij)-(Im): [ka] (In the formula, R 1 , R4 , R 5 , R 11 , Z 3 , and R 25 is as defined herein; Y 1 ~Y 3 are each independently N or CR 27 and R 24 and R 27 each independently represents H, optionally substituted C (1~6) -alkyl, optionally substituted fluoroalkyl, and halogen It can be represented by one of the following:

[0077] In some embodiments of any one of Formulas (Ie)-(Im), Z 3 is selected from —O—, —CHO—, —OCH—, optionally substituted —OCH-heteroaryl, optionally substituted —OCH-aryl, optionally substituted —CHO-heteroaryl, and optionally substituted —CHO-aryl.

[0078] In some embodiments, Z 3 teeth, [ka] is selected from.

[0079] In some embodiments of any one of Formulas (Ie)-(Im), Z 3 is -C(R 22 )2-, optionally substituted alkyl, optionally substituted amide, optionally substituted sulfonamide, optionally substituted urea, and optionally substituted thiourea. In some embodiments, Z 3 is -CH-. In some embodiments, Z 3 is -CH2CH2-. In some embodiments, Z 3 is -CH2CH2CH2-. In some embodiments, Z3 -NHSO2-(C 1~3 -alkyl). In some embodiments, Z 3 -N(Ac)-(C 1~3 -alkyl).

[0080] In some embodiments of any one of Formulas (Ie)-(Im), Z 3 -S- and NR 26 -(However, R 26 is selected from H and optionally substituted (C1-C3) alkyl.

[0081] In some embodiments of Formula (Ik), Y 1 ~Y 3 At least one of Y is N. 1 ~Y 3 At least two of the following are N. In some cases, Y 1 and Y 3 is N and Y 2 is CR 25 In some cases, Y 1 and Y 2 is N and Y 3 is CR 25 In some cases, Y 1 and Y 2 is CR 25 and Y 3 is N. In some cases, R 25 is H. In some cases, R 25 is C (1~3) -Alkyl or C (1~3) -fluoroalkyl. Optionally, the fluoroalkyl is CF3.

[0082] In certain embodiments of any one of formulas (Ie)-(Im), R 1 is OH.

[0083] In some embodiments of any one of Formulas (Ie)-(Im), R 4 and R 5 are H. In some cases, R4 and R 5 At least one of R is a pro-moiety. 4 and R 5 are cyclically linked to form a promoiety (eg, as described herein).

[0084] In certain embodiments, the compound of formula (Ie) has the following structure: [ka] is selected from one of the following:

[0085] In some embodiments, the ASGPR binding moiety (X) of a compound of the present disclosure has the formula (In′): [ka] (In the formula, R 1 , R 4 , R 5 , and Z 1 is as defined herein; Y 1 and Y 2 are each independently -O-, -S-, or NR 28 - and -C(R 22 )2 is selected, R 28 is H, optionally substituted (C1-C6) alkyl, and —C(O)R 22 is selected from Each R 22 are independently selected from H, halogen, and optionally substituted (C1-C6) alkyl; Ring B is a 5- or 6-membered optionally substituted cyclic group. In some embodiments of formula (In′), Y 1 is attached to the sugar ring via an alpha configuration. In some embodiments of formula (In′), Y 1 is attached to the sugar ring via a beta configuration.

[0086] In some embodiments, the ASGPR binding moiety (X) of a compound of the present disclosure has the formula (In): [ka] (In the formula, R 1 , R 4 , R 5 , and Z 1 is as defined herein; Y 1 and Y 2 are each independently -O-, -S-, or -NR 28 , and -C(R 22 )2 is selected, R 28 is H, optionally substituted (C1-C6) alkyl, and —C(O)R 22 is selected from Each R 22 are independently selected from H, halogen, and optionally substituted (C1-C6) alkyl; Ring B is a 5- or 6-membered optionally substituted cyclic group. It can be expressed as:

[0087] In some embodiments of formula (In) and (In′), Y 1 is O. In some cases, Y 1 is S. In some cases, Y 1 Ha-NR 28 - In some cases, Y 1 -C(R 22 )2, and each R 22 is H.

[0088] In some embodiments of formula (In) and (In′), Y 2 Ha-NR 28 -(However, R 28 is H). In some cases, Y 2 Ha-NR 28 -(However, R 28 HA-C(O)R 22 In some cases, R 22 is methyl.

[0089] In some embodiments of formula (In) and (In'), ring B is a 5- or 6-membered heterocycle. Optionally, ring B is a 5-membered heterocycle. Optionally, ring B is a 6-membered heterocycle.

[0090] In some embodiments of formula (In) and (In′), Z 1 is Z 11 (However, Z 11 -O-, -S-, NR 21 - and -C(R 22 )2) is selected from Z 1 is -O-. In some cases, Z 1 is -S-. In some cases, Z 1 is NR 21 -(However, R 21 is H). In some cases, Z 1 -C(R 22 )2 (However, each R 22 is H).

[0091] In some embodiments of formula (In) and (In′), Z 1 is an optionally substituted Z 11 -heteroaryl or optionally substituted Z 11 In some embodiments, Z 1 is CH2-heteroaryl or CH2-aryl. In some embodiments of formula (In) and (In'), Z 1 is an optionally substituted amide. In some embodiments of formula (In) and (In′), Z 1 is an optionally substituted sulfonamide. In some embodiments of formula (In) and (In′), Z 1 is an optionally substituted urea or an optionally substituted thiourea.

[0092] In some embodiments, the compounds of formula (In) and (In′) have the following structure: [ka] It has one of the following.

[0093] In certain embodiments of any one of formulas (Ia) through (In), n is 1 and L is Z 1 In certain embodiments, the linker L comprises a linear linker having a main chain of 20 or more consecutive atoms, e.g., 25 or more consecutive atoms, or 30 or more consecutive atoms, and in some cases up to 100 consecutive atoms, covalently linking X to Y (or Z) via a chain of 20 to 50 consecutive atoms. 1 In certain embodiments, the linker L separates X and Y (or Z) by a chain of 21 to 50 consecutive atoms, by a chain of 22 to 50 consecutive atoms, by a chain of 23 to 50 consecutive atoms, by a chain of 24 to 50 consecutive atoms, by a chain of 25 to 50 consecutive atoms, by a chain of 26 to 50 consecutive atoms, by a chain of 27 to 50 consecutive atoms, by a chain of 28 to 50 consecutive atoms, or by a chain of 29 to 50 consecutive atoms. 1 In certain embodiments, the linker L separates X and Y (or Z) by a chain of 30 to 60 contiguous atoms. 1 In certain embodiments, the linker L separates X and Y (or Z) by a chain of 31 to 60 contiguous atoms. 1 In certain embodiments, the linker L separates X and Y (or Z) by a chain of 32 to 60 contiguous atoms. 1 In certain embodiments, the linker L separates X and Y (or Z) by a chain of 33 to 60 contiguous atoms. 1 In certain embodiments, the linker L separates X and Y (or Z) by a chain of 34 to 60 contiguous atoms. 1 In certain embodiments, the linker L separates X and Y (or Z) by a chain of 35 to 50 contiguous atoms. 1 In certain embodiments, the linker L separates X and Y (or Z) by a chain of 36 to 50 contiguous atoms. 1In certain embodiments, the linker L separates X and Y (or Z) by a chain of 41 to 50 contiguous atoms. 1 In certain embodiments, the linker L separates X and Y (or Z) by a chain of 46 to 50 contiguous atoms. 1 ) and separate.

[0094] In certain embodiments of any one of formulas (Ia) through (In), n is 2 or greater and L is a linking moiety Z 1 is a branched linker that covalently links two or more X moieties to Y via

[0095] In certain embodiments of any one of formulas (Ia) through (In), n is 2 or greater and each branched chain of L is such that each X moiety is Z 1 Optionally, each branched chain of L comprises a linear linker of 14 to 50 consecutive atoms, e.g., 14 to 45, 14 to 40, 14 to 35, or 14 to 30 consecutive atoms. Optionally, each branched chain of L comprises a linear linker of 14 to 30 consecutive atoms, e.g., 14 to 29, 14 to 28, 14 to 27, 14 to 26, 14 to 25, 14 to 24, 14 to 23, 14 to 22, 14 to 21, or 14 to 20 consecutive atoms. Optionally, L comprises a linear linker of 14 or more consecutive atoms, e.g., 15 or more consecutive atoms, 16 or more consecutive atoms, or 17 or more consecutive atoms, optionally up to 50 consecutive atoms, covalently linked to a branch point of the linker L via a linker. Optionally, each branched chain of L comprises a linear linker of 14 to 50 consecutive atoms, e.g., 14 to 45, 14 to 40, 14 to 35, or 14 to 30 consecutive atoms. Optionally, each branched chain of L comprises a linear linker of 14 to 30 consecutive atoms, e.g., 14 to 29, 14 to 28, 14 to 27, 14 to 26, 14 to 25, 14 to 24, 14 to 23, 14 to 22, 14 to 21, or 14 to 20 consecutive atoms. Optionally, L comprises a linear linker of 14 or more consecutive atoms, e.g., 14 to 29, 14 to 28, 14 to 27, 14 to 26, 14 to 25, 14 to 24, 14 1 Optionally, L comprises more than 14 consecutive atoms covalently linked to the branch points of the linker (through a group). 1 Optionally, L may be a group consisting of 15 consecutive atoms separating the group from the branch point of L. 1 Optionally, L may be a group consisting of 16 consecutive atoms separating the group from the branch point of L. 1 Optionally, L may be a group consisting of 17 consecutive atoms separating the group from the branch point of L. 1Optionally, L may be a group consisting of 18 consecutive atoms separating the group from the branch point of L. 1 Optionally, L may be a group consisting of 19 consecutive atoms separating the group from the branch point of L. 1 Optionally, L may be a group consisting of 20 consecutive atoms separating the group from the branch point of L. 1 The group comprises a linear linker of 20 or more consecutive atoms separating the L group from the branch point.

[0096] In certain embodiments of any one of formulas (Ia) through (In), n is 2 and L is Z of each X. 1 Branched linkers having 14 or more consecutive atoms separating the group from the branch point of L are included.

[0097] In certain embodiments of any one of formulas (Ia) through (In), n is 3 and L is Z of each X. 1 Branched linkers having 14 or more consecutive atoms separating the group from the branch point of L are included.

[0098] In certain embodiments of any one of Formulas (Ia)-(In), the linker L is of Formula (II) (eg, as described herein).

[0099] In certain embodiments of any one of Formulas (Ia), (Ib), or (Id) through (In), R 1 is OH. In certain other cases, R 1 is -OC(O)R. In some cases, R 1 is —C(O)NHR, where R is an optionally substituted alkyl. In some cases, R terminates in an alkenyl or alkynyl group. In other certain cases, R 1 is an optionally substituted triazole. Optionally, the triazole has the following structure: [ka] It is of the type.

[0100] In certain embodiments of (Ia)-(Ic), R 2 is —NHCOCH. In certain other embodiments, R 2 is —NHCOCF 3 . In certain other embodiments, R 2 is -NHCOCH2CF3. In some cases, R 2 is —OH. In certain other cases, R 2 is an optionally substituted triazole. Optionally, the triazole has the following structure: [ka] It is of the type.

[0101] In certain embodiments, R 1 or R 2 When is a substituted triazole, the triazole is a 1,2,3-triazole and the substituent is at the 4- or 5-position. Optionally, the substituent on the triazole moiety can be an optionally substituted (C 1~6 ) alkyl, optionally substituted (C 1~6 ) alkoxy, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted alkaryl, and optionally substituted alkyheteroaryl. It will be understood that the triazole moiety may include any convenient substituent. See, for example, the triazole moieties disclosed in Mamidayala et al., J. Am. Chem. Soc. 2012, 134, 1978-1981.

[0102] In certain embodiments of any one of formulas (Ia)-(In), R 4 and R 5 At least one of R is a promoiety. Optionally, the promoiety is an ester. In certain embodiments, the ester is of formula -OCOCH, -OCOCH(CH), or -OCOC(CH). Optionally, R 4 and R 5At least one of R is of formula -COCH3, -COCH(CH3)2, or -COC(CH3)3. 4 and R 5 At least one of R is of the formula -CHOCOC(CH). 4 is a pro component and R 5 is H. In certain other cases, R 5 is H and R 4 is a pro component. In some cases, R 4 and R 5 Both are pro-components. 4 and R 5 are linked in a ring to form a pro moiety. 4 and R 5 are linked in a ring to form a ring of formula (Io) or (Ip): [ka] (In the formula, R 1 ~R 3 and Y 4 is as defined herein) Forming a pro component of.

[0103] In certain embodiments of any one of formulas (Ia)-(In), R 4 and R 5 Both are H.

[0104] In certain embodiments of Formula (I), n is 2 or 3 and X has the following structure: [ka] (In the formula, R 5 and R 23 are independently H or (C 1~3 ) alkyl) is selected from one of the following:

[0105] In certain embodiments of Formula (I), n is 1, 2, or 3 and X has the following structure: [ka] (In the formula, R 4 and R 5 are independently H or a pro-moiety, or R 5 and R 4 are linked in a ring to form a pro moiety, n1 and n2 each independently represent an integer of 1 to 6, and Y 4 is an appropriate counterion) In some embodiments, Y 4 is sodium.

[0106] In certain embodiments of Formula (I), n is 1, 2, or 3, and X has the following structure: [ka] is selected from one of the following:

[0107] In certain embodiments of Formula (I), n is 1, 2, or 3 and X has the following structure: [ka] is.

[0108] In certain embodiments of Formula (I), n is 1, 2, or 3 and X has the following structure: [ka] is.

[0109] In certain embodiments of Formula (I), n is 1 and X is [ka] is.

[0110] In certain embodiments of any one of formulas (Ia) to (Ip), -Z 1 -Ha-L 1-moiety (e.g., of a linker of any of formula (II), (IIa), or (IIb) described herein). In some embodiments, the subject compounds are [ka] (In the formula, o, p, q, r, s, t, u, v, w, x, y, z, and z1 each independently represent an integer of 1 to 6.) Selected from -Z 1 -L 1 - group.

[0111] In certain embodiments, the Z 1 -L 1 -The base is [ka] and o is 1 or 2.

[0112] In certain embodiments, the Z 1 -L 1 -The base is [ka] and each R 22 is H and p is 1 or 2.

[0113] In certain embodiments, the Z 1 -L 1 -The base is [ka] (wherein q is 1 to 3) is.

[0114] In certain embodiments, the Z 1 -L 1 -The base is [ka] (wherein r is 1 to 3) is.

[0115] In certain embodiments, the Z 1 -L 1 -The base is [ka] (wherein r is 1 to 3) is.

[0116] In certain embodiments, the Z 1 -L 1 -The base is [ka] (wherein s and t each independently represent 1 to 3) is.

[0117] In certain embodiments, the Z 1 -L 1 -The base is [ka] (wherein u is 1 to 3) is.

[0118] In certain embodiments, the Z 1 -L 1 -The base is [ka] (wherein v and w each independently represent 1 to 3). is.

[0119] In certain embodiments, the Z 1 -L 1 -The base is [ka] (wherein x is 0 to 3) is.

[0120] In certain embodiments, the Z 1 -L 1-The base is [ka] (wherein y is 1 to 3) is.

[0121] In certain embodiments, the Z 1 -L 1 -The base is [ka] (In the formula, R 21 is H and z is 1 to 4) is.

[0122] In certain embodiments, the Z 1 -L 1 -The base is [ka] (In the formula, R 21 is H and z1 is 1 to 4) is.

[0123] In certain embodiments, the subject compound is [ka] Selected from -Z 1 Contains the -L- group.

[0124] 5.1.1. Exemplary ASGPR Ligands Exemplary moieties that bind to ASGPR and synthons that can be utilized to prepare compounds of the present disclosure that contain an ASGPR ligand of interest are provided in Tables 1-4.

[0125] In certain embodiments, the compound of formula (Ib) is a compound shown in Table 1. [Table 1]

[0126] In certain embodiments, the compound of formula (Ic) is a compound shown in Table 2. [Table 2]

[0127] In certain embodiments, the compound of formula (Id) is a compound shown in Table 3. [Table 3]

[0128] In certain embodiments, the compound of formula (Id) is a compound shown in Table 4. [Table 4]

[0129] Exemplary building blocks that can be used to prepare compounds of the present disclosure, including the desired ASGPR ligand (X), are shown in Table 5. [Table 5-1] [Table 5-2] [Table 5-3] [Table 5-4] [Table 5-5]

[0130] In some embodiments of the building blocks of the ASGPR ligands (X) that can be used to prepare the compounds of the present disclosure, R 3 is H, so that the ASGPR ligand (X) contains a CH2 at position 1 and R 2 is the connecting part Z 1Exemplary building blocks that can be used to prepare compounds of the present disclosure containing the desired ASGPR ligand (X) are shown in Table 6. [Table 6-1] [Table 6-2] [Table 6-3] [Table 6-4] [Table 6-5]

[0131] In some embodiments, the building blocks of the ASGPR ligands (X) that can be used to prepare the compounds of the present disclosure are bicyclic structures. Exemplary building blocks for use in preparing compounds of the present disclosure that contain the desired ASGPR ligands (X) are shown in Table 7. [Table 7-1] [Table 7-2]

[0132] Prodrugs Aspects of the present disclosure include prodrugs of any of the ASGPR-binding moieties described herein that are incorporated into the compounds and conjugates of the present disclosure.

[0133] The term "prodrug" refers to a substance that is converted into a drug in vivo by some physiological or chemical process (e.g., a prodrug is converted into the desired drug form when at physiological pH).

[0134] Prodrug forms of any of the ASGPR binding moieties described herein may be useful because they may offer certain therapeutic benefits, for example, as a result of increasing the half-life of the resulting compound or conjugate in the body or reducing the effective dosage required.

[0135] This may be useful in some cases because a prodrug may be easier to administer than the parent drug. For example, a prodrug may be bioavailable by oral administration while the parent drug may not. The prodrug may also have improved solubility in pharmacological compositions over the parent drug.

[0136] Prodrug derivatives of ASGPR binding moieties generally contain a promoiety substituent at an appropriate labile site in the compound, the promoiety being a group that is removed by enzymatic or chemical reaction when the prodrug is converted to a drug in vivo.

[0137] In some embodiments, the promoiety is a group attached to a hydroxy group of the compound or drug via an ester bond.

[0138] In some embodiments, prodrug derivatives of one or more hydroxyl groups of the sugar ring may be incorporated into the compound. For example, an ester promoiety may be incorporated into one or more hydroxyl groups at the 3- and / or 4-position of a sugar (e.g., as described herein). In some embodiments, the hydroxyl groups at the 3- and 4-positions of the sugar are cyclically linked to form a promoiety (e.g., as described herein).

[0139] Linker The terms "linker," "linking moiety," and "linking group" are used interchangeably and refer to a linking moiety that covalently links two or more moieties or compounds, such as ligands and other moieties of interest. In some cases, the linker is divalent and connects two moieties. In some cases, the linker is a branched linking group that is a trivalent or higher polyvalent linker. In some cases, the linker connecting two or more moieties has a linear or branched backbone that is 500 atoms or less (e.g., 400 atoms or less, 300 atoms or less, 200 atoms or less, 100 atoms or less, 80 atoms, 60 atoms or less, 50 atoms or less, 40 atoms or less, 30 atoms or less, or even 20 atoms or less) in length, for example, as measured between the two or more moieties. The linking moiety can be a covalent bond connecting two groups, or a straight or branched chain of 1 to 500 atoms in length, e.g., about 1, 2, 3, 4, 5, 6, 8, 10, 12, 14, 16, 18, 20, 30, 40, 50, 100, 150, 200, 300, 400, or 500 carbon atoms in length, and the linker can be straight, branched, cyclic, or a single atom. In some cases, 1, 2, 3, 4, 5 or more, 10 or more, or even more carbon atoms in the linker backbone are optionally substituted with heteroatoms, e.g., sulfur, nitrogen, or oxygen heteroatoms. In some cases, when the linker includes a PEG group, every third atom in that segment of the linker backbone is substituted with oxygen. The bond between the backbone atoms can be saturated or unsaturated, and typically, there are one, two, or no more than three unsaturated bonds in the linker backbone. The linker may include one or more substituents, such as alkyl, aryl, or alkenyl groups. The linker may include, but is not limited to, one or more of oligo(ethylene glycol), ether, thioether, disulfide, amide, carbonate, carbamate, tertiary amine, alkyl (which may be linear or branched), such as methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl), and the like.The linker backbone may include a cyclic group, such as an aryl, heterocycle, cycloalkyl group, or heterocyclic group, where two or more atoms of the cyclic group, such as two, three, or four atoms, are included in the backbone.

[0140] In some embodiments, the "linker" or linking moiety is derived from a molecule having two reactive ends, one reactive end for conjugation to a moiety of interest (Y), e.g., a biomolecule (e.g., an antibody), and the other reactive end for conjugation to a moiety that binds to an ASGPR cell surface receptor (designated as X). When Y is a polypeptide, the polypeptide-binding reactive end of the linker is a site that is capable of conjugation to the polypeptide, as the case may be, via a cysteine ​​thiol or lysine amine group on the polypeptide, and thus may be a thiol-reactive group, e.g., maleimide or dibromomaleimide, or as defined herein, or an amine-reactive group such as an active ester (e.g., perfluorophenyl ester or tetrafluorophenyl ester), or as defined herein.

[0141] In certain embodiments of the formulas described herein, the linker L comprises one or more straight-chain or branched-chain carbon moieties and / or polyether (e.g., ethylene glycol) moieties (e.g., repeating units of -CHCHO-), and combinations thereof. In certain embodiments, these linkers optionally have amide, urea, or thiourea, carbamate, ester, amino, ether, thioether, sulfhydryl, heteroaryl, or other heterofunctional bonds. In certain embodiments, the linker comprises one or more carbon atoms, nitrogen atoms, sulfur atoms, oxygen atoms, and combinations thereof. In certain embodiments, the linker comprises one or more ether bonds, thioether bonds, amine bonds, amide bonds, carbon-carbon bonds, carbon-nitrogen bonds, carbon-oxygen bonds, carbon-sulfur bonds, and combinations thereof. In certain embodiments, the linker comprises a linear structure. In certain embodiments, the linker comprises a branched structure. In certain embodiments, the linker comprises a cyclic structure. Optionally, the linker includes one or more heteroaryl ring structures, such as a triazole, for example, a 1,2,3-triazole.

[0142] In certain embodiments, the length of L is about 10 Å to about 20 Å. In certain embodiments, the length of L is about 15 Å to about 20 Å. In certain embodiments, the length of L is about 15 Å. In certain embodiments, the length of L is about 16 Å. In certain embodiments, the length of L is about 17 Å.

[0143] In certain embodiments, L is a linker of about 5 Å to about 500 Å. In certain embodiments, L is about 10 Å to about 400 Å. In certain embodiments, L is about 10 Å to about 300 Å. In certain embodiments, L is about 10 Å to about 200 Å. In certain embodiments, L is about 10 Å to about 100 Å. In certain embodiments, L is about 10 Å to about 20 Å, about 20 Å to about 30 Å, about 30 Å to about 40 Å, about 40 Å to about 50 Å, about 50 Å to about 60 Å, about 60 Å to about 70 Å, about 70 Å to about 80 Å, about 80 Å to about 90 Å, or about 90 Å to about 100 Å. In certain embodiments, L is a linker of about 5 Å to about 500 Å, wherein L comprises an optionally substituted arylene linked to X, an optionally substituted heteroarylene linked to X, an alkylene group linked to X, or a heteroatom linked to X. In certain embodiments, L is a linker of about 10 Å to about 500 Å, wherein the linker comprises an optionally substituted arylene linked to X, or an optionally substituted heteroarylene linked to X, an alkylene group linked to X, or a heteroatom linked to X. In certain embodiments, L is a linker of about 10 Å to about 400 Å, wherein the linker comprises an optionally substituted arylene linked to X, or an optionally substituted heteroarylene linked to X, an alkylene group linked to X, or a heteroatom linked to X. In certain embodiments, L is a linker of about 10 Å to about 200 Å, wherein the linker comprises an optionally substituted arylene linked to X, or an optionally substituted heteroarylene linked to X, an alkylene group linked to X, or a heteroatom linked to X.

[0144] In certain embodiments, the linker L connects X and Y (or Z) by a chain of 4 to 500 contiguous atoms. 1 In certain embodiments, the linker L separates X and Y (or Z) by a chain of 4 to 50 contiguous atoms. 1In certain embodiments, the linker L separates X and Y (or Z) by a chain of 6 to 50 contiguous atoms, by a chain of 11 to 50 contiguous atoms, by a chain of 16 to 50 contiguous atoms, by a chain of 21 to 50 contiguous atoms, by a chain of 26 to 50 contiguous atoms, by a chain of 31 to 50 contiguous atoms, by a chain of 36 to 50 contiguous atoms, by a chain of 41 to 50 contiguous atoms, or by a chain of 46 to 50 contiguous atoms. 1 In certain embodiments, the linker L separates X and Y (or Z) by a chain of 6 to 50 contiguous atoms. 1 In certain embodiments, the linker L separates X and Y (or Z) by a chain of 11 to 50 contiguous atoms. 1 In certain embodiments, the linker L separates X and Y (or Z) by a chain of 16 to 50 contiguous atoms. 1 In certain embodiments, the linker L separates X and Y (or Z) by a chain of 21 to 50 contiguous atoms. 1 In certain embodiments, the linker L separates X and Y (or Z) by a chain of 26 to 50 contiguous atoms. 1 In certain embodiments, the linker L separates X and Y (or Z) by a chain of 31 to 50 contiguous atoms. 1 In certain embodiments, the linker L separates X and Y (or Z) by a chain of 36 to 50 contiguous atoms. 1 In certain embodiments, the linker L separates X and Y (or Z) by a chain of 41 to 50 contiguous atoms. 1 In certain embodiments, the linker L separates X and Y (or Z) by a chain of 46 to 50 contiguous atoms. 1 ) and separate.

[0145] In certain embodiments, the linker L connects X and Y (or Z) by a chain of 4 or 5 consecutive atoms, by a chain of 6-10 consecutive atoms, by a chain of 11-15 consecutive atoms, by a chain of 16-20 consecutive atoms, by a chain of 21-25 consecutive atoms, by a chain of 26-30 consecutive atoms, by a chain of 31-35 consecutive atoms, by a chain of 36-40 consecutive atoms, by a chain of 41-45 consecutive atoms, or by a chain of 46-50 consecutive atoms. 1 ) and separate.

[0146] In certain embodiments, the linker L is a linker between X and Y (or Z 1 ) and comprises an optionally substituted arylene linked to X, an optionally substituted heteroarylene linked to X, an optionally substituted alkylene linked to X, or a heteroatom linked to X. In certain embodiments, the linker L is a chain of 5 to 500 contiguous atoms separating X and Y (or Z). 1 ) and comprises an optionally substituted arylene linked to X, an optionally substituted heteroarylene linked to X, an optionally substituted alkylene linked to X, or a heteroatom linked to X. In certain embodiments, the linker L is a chain of 7 to 500 contiguous atoms separating X and Y (or Z). 1 ) and comprises an optionally substituted arylene linked to X, an optionally substituted heteroarylene linked to X, an optionally substituted alkylene linked to X, or a heteroatom linked to X. In certain embodiments, the linker L is a chain of 10 to 500 contiguous atoms separating X and Y (or Z). 1 ), and comprises an optionally substituted arylene linked to X, an optionally substituted heteroarylene linked to X, an optionally substituted alkylene linked to X, or a heteroatom linked to X.

[0147] In certain embodiments, the linker L is a linker between X and Y (or Z 1) and comprises an alkylene, heteroatom, or optionally substituted heteroarylene linked to X. In certain embodiments, the linker L is a chain of 5 to 500 contiguous atoms separating X and Y (or Z). 1 ) and comprises an alkylene, heteroatom, or optionally substituted heteroarylene linked to X. In certain embodiments, the linker L is a chain of 7 to 500 contiguous atoms separating X and Y (or Z). 1 ) and comprises an alkylene, heteroatom, or optionally substituted heteroarylene linked to X. In certain embodiments, the linker L is a chain of 10 to 500 contiguous atoms separating X and Y (or Z). 1 ) and comprises an alkylene, heteroatom, or optionally substituted heteroarylene linked to X.

[0148] In certain embodiments, the linker L is a linker between X and Y (or Z 1 ) and comprises an optionally substituted triazole linked to X. In certain embodiments, the linker L is a chain of 5 to 500 contiguous atoms separating X and Y (or Z). 1 ) and comprises an optionally substituted triazole linked to X. In certain embodiments, the linker L is a chain of 7 to 500 contiguous atoms separating X and Y (or Z). 1 ) and comprises an optionally substituted triazole linked to X. In certain embodiments, the linker L is a chain of 10 to 500 contiguous atoms separating X and Y (or Z). 1 ) and includes an optionally substituted triazole linked to X.

[0149] In certain embodiments, the linker L is a chain of 16 to 400 contiguous atoms separating X and Y (or Z) and comprises an optionally substituted arylene linked to X, an optionally substituted heteroarylene linked to X, an optionally substituted alkylene linked to X, or a heteroatom linked to X.

[0150] The linker may be a linker between the Z of the ASGPR binding moiety (X) (e.g., as described herein). 1 It is understood that the linker may be considered to be directly attached to the Z group. In some embodiments of any of Formulas (Ia)-(Ip), the linker 1 Alternatively, the -Z group may be considered to be directly bonded to the -Z group. 1 -L 1 The - group (e.g., as described herein) may be considered part of the linking moiety connecting L to Y. The present disclosure is intended to include all such configurations of ASGPR binding moieties (X) and linkers (L).

[0151] In some embodiments of Formula (I)-(Ia), L is a group represented by Formula (II): [ka] (In the formula, L 1 and L 3 are independently linkers, and L 2 is a branched chain linking moiety, where L 1 ~L 3 together provide a linear or branched linker between X and Y, a, b, and c are independently 0 or 1; ** is X's Z 1 via L 1 represents the point of attachment to *** represents the point of attachment to Y; when n is 1, a is 1 and b is 0; If n is >1, then a is 1 and b is 1. is a linker.

[0152] In certain embodiments of the linker of formula (II), L 1 ~L 3 are each independently, -C 1~20 -Alkylene-, -NHCO-C 1~6 -Alkylene-, -CONH-C 1~6 -Alkylene-, -NHC 1~6-Alkylene-, -NHCONH-C 1~6 -Alkylene-, -NHCSNH-C 1~6 -Alkylene-, -C 1~6 -Alkylene-NHCO-, -C 1~6 -Alkylene-CONH-, -C 1~6 -Alkylene-NH-, -C 1~6 -Alkylene-NHCONH-, -C 1~6 -Alkylene-NHCSNH-, -O(CH2) p -, -(OCH2CH2) p and one or more linking moieties independently selected from -, -NHCO-, -CONH-, -NHSO2-, -SONH-, -CO-, -SO2-, -O-, -S-, monocyclic heteroaryl (e.g., 1,2,3-triazole), monocyclic aryl (e.g., phenyl, e.g., 1,4-linked phenyl or 1,3-linked phenyl), monocyclic heterocycle (e.g., a pyrrolidine-2,5-dione, piperazine, or piperidine ring, as described herein), amino acid residue (natural or non-naturally occurring amino acid residue), -NH-, and -NMe- (wherein each p is independently 1 to 50).

[0153] In certain embodiments of the linker of formula (II), L 1 ~L 3 contains repeating ethylene glycol moieties (e.g., -CH2CHO- or -OCH2CH2-). In certain cases, the linker of formula (II) above contains 1 to 25 ethylene glycol moieties, e.g., 3 to 25, 5 to 25, 7 to 25, 10 to 25, 15 to 25, 17 to 25, 20 to 25, or 22 to 25 ethylene glycol moieties. In some cases, the linker of formula (II) above contains 3 or more ethylene glycol moieties, e.g., 5 or more, 7 or more, 10 or more, 15 or more, 20 or more, or even more ethylene glycol moieties.

[0154] In certain embodiments of the linker of formula (II), L 1 ~L 3In some cases, the linker comprises one or more 1,2,3-triazole linking moieties. In certain cases, the one or more 1,2,3-triazole moieties have the following structure: [ka] (wherein w1, u1, and q1 are independently selected from one of 1 to 25 (eg, 1 to 12, for example, 1 to 6)).

[0155] In certain embodiments of the linker of formula (II) above, n is 1 and therefore b is 0, and the linker has formula (IIa): [ka] (In the formula, L 1 and L 3 are independently a linker (e.g., as described herein), provided that L 1 ~L 3 together provide a linear linker between X and Y, a is "1" c is 0 or 1, ** is X's Z 1 via L 1 represents the point of attachment to *** indicates the point of attachment to Y) is a linker.

[0156] In certain embodiments of the linker of formula (IIa) above, the linear linker is Z 1 In certain embodiments of Formula (IIa), the linear linker has a main chain of 20 or more consecutive atoms, e.g., 25 or more consecutive atoms, or 30 or more consecutive atoms, and in some cases, up to 100 consecutive atoms, covalently linking X to Y (or Z) via a chain of 20 to 50 consecutive atoms. 1In certain embodiments, the linear linker separates X and Y (or Z) by a chain of 21 to 50 consecutive atoms, by a chain of 22 to 50 consecutive atoms, by a chain of 23 to 50 consecutive atoms, by a chain of 24 to 50 consecutive atoms, by a chain of 25 to 50 consecutive atoms, by a chain of 26 to 50 consecutive atoms, by a chain of 27 to 50 consecutive atoms, by a chain of 28 to 50 consecutive atoms, or by 29 to 50 consecutive atoms. 1 In certain embodiments of Formula (IIa), the linear linker separates X and Y (or Z) by a chain of 30 to 60 contiguous atoms. 1 In certain embodiments, the linear linker separates X and Y (or Z) by a chain of 31 to 60 contiguous atoms. 1 In certain embodiments, the linear linker separates X and Y (or Z) by a chain of 32 to 60 contiguous atoms. 1 In certain embodiments, the linear linker separates X and Y (or Z) by a chain of 33 to 60 contiguous atoms. 1 In certain embodiments, the linear linker separates X and Y (or Z) by a chain of 34 to 60 contiguous atoms. 1 In certain embodiments, the linear linker L separates X and Y (or Z) by a chain of 35 to 50 consecutive atoms. 1 In certain embodiments, the linear linker L separates X and Y (or Z) by a chain of 36 to 50 consecutive atoms. 1 In certain embodiments, the linear linker L separates X and Y (or Z) by a chain of 41 to 50 consecutive atoms. 1 In certain embodiments, the linear linker L separates X and Y (or Z) by a chain of 46 to 50 consecutive atoms. 1 ) and separate.

[0157] In certain other embodiments of Formula (II), n is 2 or greater and L 1 ~L 3 together provide a branched linker between X and Y.

[0158] In certain embodiments of Formula (II), n is 2 or greater and L 2 teeth, [ka] (wherein each x and y independently represents 1 to 10) is selected from.

[0159] In certain embodiments of Formula (II), L 1 ~L 2 comprises a main chain of 14 or more consecutive atoms between X and the branch atom, for example, 14 to 50, 14 to 40, 14 to 35, or 14 to 30 consecutive atoms between X and the branch atom.

[0160] In certain embodiments of Formula (II) or (IIa), L 3 comprises a main chain of 10 to 80 consecutive atoms, for example, 12 to 70, 12 to 60, or 12 to 50 consecutive atoms.

[0161] In certain embodiments of Formula (II) or (IIa), L 3 is C 10 ~C 20 -Alkylene (e.g., C 12 -alkylene), or -(OCH2CH2) p -(wherein p is 1 to 25, e.g., 3 to 25, 5 to 24, 7 to 25, 10 to 25, 15 to 25, or 20 to 24).

[0162] In certain embodiments, L has formula (IIb): [ka] (In the formula, Each L 1 ~L 5 are independently, and both are Z 1 and Y, a, b, c, d, and e are each independently 0, 1, or 2; ** is X's Z 1 via L 1 represents the point of attachment to *** represents the point of attachment to Y; when n is 1, a is 1 and c is 0; If n>1, then a is 1 and c is 1) It is L.

[0163] In certain embodiments of the linker of formula (IIb) above, L 1 ~L 5 are each independently, -C 1~20 -Alkylene-, -NHCO-C 1~6 -Alkylene-, -CONH-C 1~6 -Alkylene-, -NHC 1~6 -Alkylene-, -NHCONH-C 1~6 -Alkylene-, -NHCSNH-C 1~6 -Alkylene-, -C 1~6 -Alkylene-NHCO-, -C 1~6 -Alkylene-CONH-, -C 1~6 -Alkylene-NH-, -C 1~6 -Alkylene-NHCONH-, -C 1~6 -Alkylene-NHCSNH-, -O(CH2) p -, -(OCH2CH2) p and one or more linking moieties independently selected from -, -NHCO-, -CONH-, -NHSO2-, -SONH-, -CO-, -SO2-, -O-, -S-, monocyclic heteroaryl (e.g., 1,2,3-triazole), monocyclic aryl (e.g., phenyl, e.g., 1,4-linked phenyl or 1,3-linked phenyl), monocyclic heterocycle (e.g., a pyrrolidine-2,5-dione, piperazine, or piperidine ring, as described herein), amino acid residue (natural or non-naturally occurring amino acid residue), -NH-, and -NMe- (wherein each p is independently 1 to 50).

[0164] In certain embodiments of Formula (IIb), -(L 1 ) a- includes an optionally substituted alkyl or ethylene glycol linking moiety. 1 is optionally replaced by -C 1~6 In certain cases, L 1 contains an ethylene glycol linking moiety.

[0165] In certain embodiments of Formula (IIb), L 1 are independent, -C 1~6 -Alkylene-, -(CH2CH2O) t -, -C 1~6 -Alkylene-NR 4 CO-, -C 1~6 -alkylene CONH-, or OCH2, where t is 1 to 20; R 4 is independently selected from H and optionally substituted (C1-C6) alkyl. In certain cases, L 1 Ha-C 1~6 -Alkylene-, for example, -C 1~3 -alkylene-. In certain cases, L 1 Ha-(CH2CH2O) t -, where t is 1 to 20, e.g., 1 to 15, 1 to 10, 1 to 8, 1 to 6, or 1 to 4. In certain cases, L 1 Ha-C 1~6 -Alkylene-NR 4 CO-. In certain cases, L 1 Ha-C 1~6 -alkyleneCONH-. In certain cases, L 1 is OCH2.

[0166] In some embodiments of Formula (IIb), one or more L 1 are independently -CH2O-, -(CH2CH2O) t -, -NR 4 CO-, -C 1~6 -alkylene-, [ka] (In the formula, R 13is H, halogen, OH, optionally substituted (C1-C6) alkyl, optionally substituted (C1-C6) alkoxy, COOH, NO2, CN, NH2, -N(R 21 )2, -OCOR 21 , -COOR 21 , -CONHR 21 , and -NHCOR 21 is selected from Each r is independently 0 to 20, and L 1 any of the moieties is optionally further substituted) is.

[0167] In certain embodiments of Formula (IIb), L 2 are independently, -NR 4’ CO-C 1~6 -Alkylene-, -CONR 4’ -C 1~6 alkylene, [ka] -OCH2- and -(OCH2CH2) q wherein q is 1 to 10, u is 0 to 10, w is 1 to 10, and R 4’ is independently selected from H and optionally substituted (C1-C6) alkyl. In certain cases, L 2 Ha-NR 4’ CO-C 1~6 -alkylene-. In certain cases, L 2 -CONR 4’ -C 1~6 - alkylene.

[0168] In certain cases, L 2 teeth [ka] (wherein w is 1 and u is 0 or 1) is.

[0169] In certain cases, L 2 teeth [ka] (wherein w is 1 and u is 0 or 1) is.

[0170] In certain cases, L 2 teeth [ka] (wherein w is 1, u is 0 or 1, and q is 1) is.

[0171] In certain cases, L 2 teeth [ka] (wherein u is 0 or 1) is.

[0172] In certain cases, L 2 teeth [ka] is.

[0173] In certain embodiments, L 2 is —OCH—. In certain other embodiments, L 2 is (OCH2CH2) q - and q is 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, or 1 to 2. In certain cases, q is 2 to 8, for example, 2 to 6, 4 to 6, or 2 to 4.

[0174] In certain embodiments of Formula (IIb), L 4 is absent or independently, -C 1~6 -Alkylene-, -(CH2CH2O) t -, -C 1~6 -Alkylene-NHCO-, -C 1~6-alkyleneCONH-, or OCH2, where t is 1 to 20. 4 does not exist. In certain cases, L 4 Ha-C 1~6 -alkylene-. In certain cases, L 4 Ha-(CH2CH2O) t -, and t is 1 to 20, e.g., 1 to 15, 1 to 12, 1 to 10, 1 to 8, 1 to 6, 1 to 4, or 1 to 3. In certain cases, L 4 Ha-C 1~6 -alkylene-NHCO-. In certain cases, L 4 Ha-C 1~6 -alkyleneCONH-. In certain cases, L 4 is OCH2.

[0175] In some embodiments of the subject compounds, n is 1 and L in formula (IIb) 3 does not exist.

[0176] In certain embodiments of the subject compounds, n is 2 or greater and L of formula (IIb) 3 is a branched chain linking moiety.

[0177] Thus, in some embodiments of formula (IIb), L 3 is a branched linking moiety, e.g., a divalent or trivalent linking moiety. For example, L 3 The linking moiety has the following general formula: [ka] It may be one of the linking moieties.

[0178] In some embodiments of Formula (IIb), the branched linking moiety may be a higher valency linking moiety and has the general formula: [ka] (Wherein, any two L 3The groups may be directly linked or attached via an optional linear linking moiety (e.g., as described herein). etc.

[0179] In some embodiments of Formula (IIb), the branched linking moiety may include one, two, or more L 3 and optionally containing linking moieties, each of which is a trivalent moiety that, when linked together, provides multiple branching points for covalent attachment of ligands and has the general formula: [ka] (wherein t is 0 to 500, for example, 0 to 100, 0 to 20, or 0 to 10) It may be represented by:

[0180] In some embodiments, the branched chain linking moiety (e.g., L 3 ) includes one or more amino acid residues (e.g., Asp, Lys, Orn, Glu), N-substituted amide (-N(-)C(=O)-), tertiary amino, polyol (e.g., O-substituted glycerol), and the like.

[0181] In some embodiments of Formula (IIb), one or more L 3 teeth, [ka] wherein each x and y is independently 1 to 10, e.g., 1 to 6, 1 to 3, e.g., 1 or 2. In some cases, each x is 1, 2, or 3, e.g., 2. is a branched chain moiety selected from

[0182] In some embodiments of Formula (IIb), L 5 is -CH2O-, -(CH2CH2O) t -, -NR 4 CO-, -C 1~6 -alkylene-, [ka] (In the formula, R 13 is H, halogen, OH, optionally substituted (C1-C6) alkyl, optionally substituted (C1-C6) alkoxy, COOH, NO2, CN, NH2, -N(R 21 )2, -OCOR 21 , -COOR 21 , -CONHR 21 , and -NHCOR 21 is selected from Each r is independently 0 to 20, and L 5 any of the moieties is optionally further substituted) is selected from.

[0183] In certain cases, L 5 is -CH2O-. In certain cases, L 5 Ha-(CH2CH2O) t -, and t is 1 to 20, e.g., 1 to 15, 1 to 12, 1 to 10, 1 to 8, 1 to 6, or 1 to 4. In certain cases, L 5 Ha-NR 4 CO- and R 4 is H or optionally substituted (C1-C6) alkyl. In certain cases, L 5 Ha-C 1~6 -alkylene-.

[0184] In certain cases, L 5 teeth [ka] (wherein r is 0 to 20, for example, 0 to 15, 0 to 10, 0 to 8, or 0 to 5). is.

[0185] In certain cases, L 5 teeth [ka] wherein each r is independently 0 to 20, e.g., 0 to 15, 0 to 10, 0 to 8, or 0 to 5; R 13 is H or optionally substituted (C1-C6) alkyl is.

[0186] In certain cases, L 5 teeth [ka] (wherein r is 0 to 20, e.g., 0 to 15, 0 to 10, 0 to 8, or 0 to 5; R 13 is H or optionally substituted (C1-C6) alkyl is.

[0187] In certain cases, L 5 teeth [ka] (wherein r is 0 to 20, e.g., 0 to 15, 0 to 10, 0 to 8, or 0 to 5; R 13 is H or optionally substituted (C1-C6) alkyl is.

[0188] In certain cases, L 5 teeth [ka] (wherein r is 0 to 20, e.g., 0 to 15, 0 to 10, 0 to 8, or 0 to 5; R 13 is H or optionally substituted (C1-C6) alkyl is.

[0189] In certain cases, L 5 teeth [ka] (wherein r is independently 0 to 20, for example, 0 to 15, 0 to 10, 0 to 8, or 0 to 5). is.

[0190] In certain cases, L 5 teeth [ka] (wherein each r is independently 0 to 20, for example, 0 to 15, 0 to 10, 0 to 8, or 0 to 5). is.

[0191] In certain cases, L 5 teeth [ka] (wherein each r is independently 0 to 20, for example, 0 to 15, 0 to 10, 0 to 8, or 0 to 5). is.

[0192] In certain cases, L 5 teeth [ka] (wherein each r is independently 0 to 20, for example, 0 to 15, 0 to 10, 0 to 8, or 0 to 5). is.

[0193] In certain cases, L 5 teeth [ka] (wherein r is 0 to 20, for example, 0 to 15, 0 to 10, 0 to 8, or 0 to 5). is.

[0194] In some embodiments of Formula (IIb), L 5includes one or more amino acid residues (e.g., Asp, Lys, Orn, Glu), amino acid analogs, N-substituted amides (-N(-)C(=O)-), tertiary amino acids, polyols (e.g., O-substituted glycerol), etc. Amino acid analogs include, but are not limited to, unnatural amino acids, as well as other modifications known in the art. The amino acids may include L-amino acids, D-amino acids, or both, and may include any of a variety of amino acid modifications or analogs known in the art.

[0195] In some embodiments of Formula (IIb), L 1 ~L 5 is in the following units:

[0196] [ka] (In the formula, R a is (C1-C6) alkyl or substituted (C1-C6) alkyl, for example, (C1-C6) alkyl optionally substituted with an amine, a tertiary amine, optionally substituted alkoxy, optionally substituted carboxyl, optionally substituted aryl, or optionally substituted heteroaryl. Contains one or more of the following: R a It is to be understood that may be linked to the M6PR binding moiety.

[0197] In some embodiments, the linker comprises a polypeptide scaffold in which some or all of the side chain groups of its amino acid residues have been modified to bind to an ASGPR-binding moiety (e.g., as described herein). It should be understood that an ASGPR-binding moiety (e.g., as described herein) can be conjugated to amino acid residues such as Asp, Lys, Orn, Glu, and Ser of a polypeptide comprising the linker via convenient conjugation chemistry. In some embodiments, the linker comprises a polylysine polypeptide. In some embodiments, the linker comprises a polyornithine polypeptide. In some embodiments, the linker comprises a polyserine polypeptide. In some embodiments, the linker comprises a polyaspartic acid polypeptide. The polypeptide can be a randomly polymerized polymer having an average length, or a polymer of a defined length, for example, prepared in a controlled, stepwise manner. In some cases, the length of the polypeptide linker segment is 10 to 100 amino acid residues, e.g., 20 to 90 amino acid residues, or 20 to 50 amino acid residues. In some embodiments, the N- or C-terminus of the polypeptide linker segment is modified to include a linking unit to an additional M6PR binding moiety (e.g., as described herein). In some embodiments, the N- or C-terminus of the polypeptide linker segment is modified with one or more linking units (e.g., as described herein) suitable for attachment to a Y, moiety of interest.

[0198] In certain embodiments of Formula (IIb), a is 1. In certain cases, at least one of b, c, d, and e is not 0. In certain cases, b is 1 or 2. In certain cases, c is 1 or 2. In certain cases, e is 1 or 2. In certain cases, b, d, and e are independently 1 or 2. In certain cases, a, b, d, and e are each 1 and c is 0.

[0199] In certain embodiments of Formula (II), (IIa), or (IIb), the linker comprises 20 to 100 consecutive atoms, e.g., 20 to 90, 20 to 80, 20 to 70, 20 to 60, 20 to 50, 20 to 40, or 20 to 30 consecutive atoms. In certain cases, the linker comprises 25 to 100 consecutive atoms, e.g., 30 to 100, 35 to 100, 40 to 100, 45 to 100, 50 to 100, 55 to 100, 60 to 100, 65 to 100, 70 to 100, 75 to 100, 80 to 100, 85 to 100, 90 to 100, or 95 to 100 consecutive atoms.

[0200] In certain embodiments of Formula (II), (IIa), or (IIb), the linker comprises 25 or more consecutive atoms, e.g., 26 or more, 27 or more, 28 or more, 29 or more, or 30 or more consecutive atoms. In certain embodiments of Formula (II), (IIa), or (IIb), the linker comprises 30 or more consecutive atoms, e.g., 31 or more, 32 or more, 33 or more, 34 or more, 35 or more, 36 or more, 37 or more, 38 or more, 39 or more, 40 or even more consecutive atoms.

[0201] The present inventors have demonstrated that the ASGPR-binding compounds of the present disclosure, with specific configurations and linkers of desired valency and length, can simultaneously bind specifically to both the ASGPR and the target with high affinity and exhibit high target uptake activity. Thus, the conjugates of the present disclosure can sequester and degrade the target protein in the lysosomes of cells. For example, the ASGPR-binding moiety (e.g., Z 1 Conjugates of trivalent ASGPR-binding compounds having 14 or more atoms between the ASGPR-binding compound (e.g., Z 1These conjugates can exhibit superior cellular uptake compared to conjugates of trivalent ASGPR-binding compounds having a shorter linker (e.g., a linker of less than 14 atoms) between the 1-triazole moiety and the branch point. For example, the inventors observed that a conjugate having a 1-triazole moiety and a short bond (e.g., 6 atoms) from the ASGPR ligand to the branch point of the ligand (I-157, linker length to the branch point: 6 atoms) exhibited lower uptake activity in HepG2 cells than a conjugate having a 1-triazole moiety and a longer bond (e.g., 14 atoms) from the ASGPR ligand to the branch point (I-143, length: 14 atoms) (see, e.g., Figure 2A). Based on this finding, the present specification describes multivalent ASGPR-binding compounds having a specific range of linker lengths between the ASGPR-binding moiety and the linker branch point that desirably bind to targets and desirably internalize the bound targets into cells.

[0202] Furthermore, conjugates of trivalent ASGPR-binding compounds (e.g., compounds of Formula (I) where n=3) can exhibit superior cellular uptake activity compared to conjugates of divalent or monovalent ASGPR-binding compounds (e.g., compounds of Formula (I) where n=2 or 1). The inventors have observed that conjugate I-124 (n=3) exhibits superior uptake activity in HepG2 cells compared to divalent conjugate I-144 (n=2) (see, e.g., Figure 2B). Still further, conjugates of multivalent ASGPR-binding compounds having 12 or more atoms between the branch point of the linker and the Y moiety of interest can exhibit superior cellular uptake compared to conjugates of multivalent ASGPR-binding compounds having shorter linkers (e.g., linkers of less than 12 atoms) between the branch point of the linker and the Y moiety of interest. The present inventors have also demonstrated that conjugates of ASGPR-binding compounds having more than 12 atoms between the branch point of the linker and Y exhibit comparable uptake activity. For example, conjugates having longer linkers between the ASGPR linker and Y (e.g., the conjugate of compound I-137, which has 81 atoms between the branch point and Y, and the conjugate of compound I-129, which has 33 atoms between the branch point and Y) were observed to exhibit comparable activity to a reference conjugate (e.g., the conjugate of compound I-124, which has 12 atoms between the branch point and Y) (see, e.g., Figure 2B).

[0203] Thus, in certain embodiments where the linker of formula (II) or (IIb) is a branched linker, each branch of the linker may be Z 1In certain cases, each branched chain of the linker comprises a linear linker of 14 or more consecutive atoms covalently linking each X moiety to a branch point of the linker via a linear linker of 15 or more consecutive atoms to the branch point. In certain cases, each branched chain of the linker comprises a linear linker of 16 or more consecutive atoms to the branch point. In certain cases, each branched chain of the linker comprises a linear linker of 17 or more consecutive atoms to the branch point. In certain cases, each branched chain of the linker comprises a linear linker of 18 or more consecutive atoms to the branch point. In certain cases, each branched chain of the linker comprises a linear linker of 19 or more consecutive atoms to the branch point.

[0204] In certain embodiments of Formula (II) or (IIb), the linker is Z 1 and a linear linker covalently connecting the branch point to Y. In certain cases, the linear linker covalently connecting the branch point to Y is 12 or more consecutive atoms. In certain cases, the linear linker covalently connecting the branch point to Y is 15 or more consecutive atoms. In certain cases, the linear linker covalently connecting the branch point to Y is 20 or more consecutive atoms. In certain cases, the linear linker covalently connecting the branch point to Y is 25 or more consecutive atoms. In certain cases, the linear linker covalently connecting the branch point to Y is 30 or more consecutive atoms. In certain cases, the linear linker covalently connecting the branch point to Y is 40 or more consecutive atoms. In certain cases, the linear linker covalently connecting the branch point to Y is 50 or more consecutive atoms. In certain cases, the linear linker covalently connecting the branch point to Y is 60 or more consecutive atoms. In certain cases, the linear linker covalently connecting the branch point to Y is 70 or more consecutive atoms. In certain cases, the linear linker covalently connecting the branch point to Y is 80 or more consecutive atoms.

[0205] 5.2.1. Exemplary Linkers and Linking Moieties Exemplary linkers and linking moieties (eg, linkers and linking moieties that connect an ASGPR ligand (X) to a moiety of interest (Y)) that can be utilized in preparing compounds of the present disclosure are shown in Tables 8-10.

[0206] In certain embodiments, the linker is a linear linker or linking moiety shown in Table 8. [Table 8-1] [Table 8-2] [Table 8-3]

[0207] Table 9 includes synthetic precursors of various linker components (eg, precursors of linear and branched linkers) that can be utilized in the preparation of the subject compounds. [Table 9-1] [Table 9-2] [Table 9-3] [Table 9-4] [Table 9-5] [Table 9-6] [Table 9-7]

[0208] In certain embodiments, the linker is a branched linker or linking moiety shown in Table 10. [Table 10-1] [Table 10-2] [Table 10-3] [Table 10-4]

[0209] 5.3. Chemoselective Linking Groups In certain embodiments of Formula (I), Y is a chemoselective linking group or a precursor thereof. A chemoselective linking group is a group or functional group having a reactive functionality that allows conjugation to a compatible group of a second moiety. For example, the chemoselective linking group (or a precursor thereof) can be one of a pair of groups associated with conjugation chemistries such as azide-alkyne click chemistry, copper-free click chemistry, Staudinger ligation, tetrazine ligation, hydrazine-iso-Pictet-Spengler (HIPS) ligation, cysteine-reactive ligation chemistry (e.g., thiol-maleimide, thiol-haloacetamide, or alkyne hydrothiolation), amine-activated ester coupling, tyrosine-specific conjugation chemistry (e.g., eY-CLICK), methionine-specific conjugation chemistry (e.g., oxaziridine-based or ReACT chemistry), reductive amination, dialkyl squarate chemistry, etc.

[0210] Chemoselective linking groups that can be used to link two moieties include, but are not limited to, amino (e.g., the N-terminal amino or lysine side chain group of a polypeptide), azide, arylazide, alkynyl (e.g., ethynyl or cyclooctyne or derivatives), active ester (e.g., N-hydroxysuccinimide (NHS) ester, sulfo-NHS ester, or PFP ester or thioester), haloacetamide (e.g., iodoacetamide or bromoacetamide), chloroacetyl, bromoacetyl, hydrazide, maleimide, vinyl sulfone, 2-sulfonylpyridine, cyano-alkyne, thiol (e.g., cysteine ​​residue), disulfide or protected thiol, isocyanate, isothiocyanate, aldehyde, ketone, alkoxyamine, hydrazide, aminooxy, phosphine, HIPS hydrazinyl-indolyl group, or aza-HIPS hydrazinyl-pyrrolo-pyridinyl group, tetrazine, cyclooctene, squarate, and the like.

[0211] In some cases, the chemoselective linking group is capable of spontaneously conjugating to a compatible chemical group when the two groups are contacted under suitable conditions (e.g., copper-free click chemistry conditions). In some cases, the chemoselective linking group is capable of conjugating to a compatible chemical group when the two groups are contacted in the presence of a catalyst or other reactant (e.g., copper-catalyzed click chemistry conditions).

[0212] In some embodiments, the chemoselective linking group is a photoactive linking group. For example, upon irradiation with ultraviolet light, the diazirine group can form a reactive carbene that can insert into the C-H, N-H, and O-H bonds of a second moiety.

[0213] In some cases, Y is a reactive functional group or a precursor of a functional group capable of conjugation to a compatible group on a second moiety. For example, a carboxylic acid is a precursor to an active ester chemoselective linking group.

[0214] In certain embodiments of Formula (I), Y is a reactive moiety capable of forming a covalent bond to a polypeptide (e.g., with an amino acid side chain of the polypeptide bearing a compatible reactive group), which reactive moiety may be referred to as a chemoselective linking group.

[0215] In certain embodiments of Formula (I), Y is a thio-reactive chemoselective linking group (e.g., as described in Table 11). Optionally, Y may generate a residual moiety, Z, resulting from the covalent attachment of the thiol-reactive chemoselective linking group to one or more cysteine ​​residue(s) of a protein (e.g., an Ab).

[0216] In certain embodiments of Formula (I), Y is an amino-reactive chemoselective linking group (e.g., as described in Table 11). Optionally, Y may generate a residual moiety, Z, resulting from the covalent attachment of the amine-reactive chemoselective linking group to one or more lysine residue(s) of a protein (e.g., Ab).

[0217] Exemplary chemoselective linking groups, and their synthetic precursors, that can be adapted for use in the compounds of the present disclosure are shown in Table 11. [Table 11-1] [Table 11-2] [Table 11-3]

[0218] In Table 11, [ka] may represent the linking moiety or the point of attachment of Y to the linked X moiety.

[0219] 5.3.1. Exemplary Compounds with Chemoselective Linking Groups This disclosure includes: (1) one or more specific ASGPR ligands (X) (e.g., ligands X1-X20 in Tables 1-4, as described herein) or specific ASGPR ligands (X) (e.g., as described herein); (2) a linker comprising one or more linking moieties (e.g., any one or more of the linking moieties in Tables 8-10 described herein), and (3) a chemoselective linking group (Y), such as any one of the groups in Table 11 described herein. Compounds of formula (I) include compounds of formula (I) which may comprise:

[0220] Table 12 shows various monovalent ligand-linker compounds for use in the conjugates of the present disclosure. [Table 12]

[0221] Table 13 shows various multivalent ligand-linker compounds for use in the conjugates of the present disclosure. [Table 13]

[0222] Tables 14-17 show some exemplary ASGPR-binding compounds of the present disclosure that include a chemoselective linking group or precursor thereof. It should be understood that the present disclosure includes conjugates of each Y (e.g., as described herein) of the exemplary compounds of Tables 12-18. For example, conjugates in which the chemoselective linking group is attached to a different Y, such as a biomolecular or small molecule ligand for a target protein.

[0223] The chemoselective linking group of such compounds can be utilized to attach to another Y moiety of interest (e.g., as described below). It should be understood that any of these compounds can also be prepared a priori to include another Y moiety of interest (e.g., as described below) rather than a chemoselective linking group. In some embodiments, such compounds are referred to as conjugates, e.g., biomolecular conjugates that specifically bind to target proteins. [Table 14-1] [Table 14-2] [Table 14-3] [Table 15-1] [Table 15-2] [Table 15-3] [Table 15-4] [Table 15-5] [Table 15-6] [Table 15-7] [Table 15-8] [Table 15-9] [Table 15-10] [Table 15-11] [Table 15-12] [Table 15-13] [Table 15-14] [Table 15-15] [Table 15-16] [Table 15-17] [Table 16-1] [Table 16-2] [Table 17-1] [Table 17-2]

[0224] The present disclosure is intended to encompass any one stereoisomer of the compounds described herein. In some cases, the compounds include an enantiomer of DN-acetylgalactosamine (GalNAc), or an analog or derivative of GalNAc.

[0225] 5.3.2. Other Exemplary Compounds Table 18 shows exemplary ASGPR-binding compounds of the present disclosure that include a binding moiety or precursor thereof. [Table 18-1] [Table 18-2]

[0226] Table 19 shows exemplary trivalent ASGPR binding intermediate compounds of the present disclosure that include the X group of formula (Ie). [Table 19-1] [Table 19-2] [Table 19-3] [Table 19-4] [Table 19-5] [Table 19-6] [Table 19-7] [Table 19-8] [Table 19-9]

[0227] Table 20 shows exemplary monovalent ASGPR-binding intermediate compounds of the present disclosure that include a promoiety and an X group that is of formula (Ib). [Table 20]

[0228] Table 21 shows exemplary ASGPR-binding intermediate compounds of the present disclosure that include an X group that is of formula (In). [Table 21]

[0229] 5.4. Conjugation with a Moiety of Interest The compounds of the present disclosure may be referred to as conjugates, for example, when the moiety of interest (Y) is a molecule (e.g., as described herein). Such conjugates can be prepared by conjugation of a chemoselective linking group of any one of the compounds described herein with a compatible reactive group of molecule Y. The compatible group of molecule Y may be introduced by modification prior to conjugation or may be a group present in the molecule. Alternatively, such conjugates can be prepared de novo via modification of the starting molecule of interest Y to, for example, introduce a linker to which ligand X can be attached.

[0230] In some embodiments, the moiety of interest to which the ASGPR-binding moiety is linked is a biomolecule. In some embodiments, the moiety of interest is a biomolecule. In some embodiments, the biomolecule is selected from a peptide, a protein, a polynucleotide, a polysaccharide, a glycan, a glycoprotein, a lipid, an enzyme, an antibody, and an antibody fragment. In some embodiments, the moiety of interest Y is selected from a small molecule, a small molecule drug, a chemotherapeutic agent, a cytotoxic agent, a diagnostic agent, a dye, a fluorophore, etc.

[0231] In a preferred embodiment, the moiety of interest is a molecule that specifically binds to a target of interest, i.e., a target-binding moiety. In such cases, the conjugate of the present disclosure can internalize and / or degrade the target after the target is non-covalently bound to the conjugate. The present inventors have demonstrated that the conjugate of the present disclosure, which has a specific configuration of an ASGPR-binding moiety with a desired affinity and a linker with a desired valency and length, can simultaneously specifically bind to both the ASGPR and the target with high affinity. Therefore, the conjugate of the present disclosure can sequester the target protein in the lysosome of the cell and degrade the target protein.

[0232] In some embodiments, the moiety of interest is a molecule that does not bind to an extracellular target, but rather is itself a molecule that is desirable for intracellular delivery, hi some embodiments, the moiety of interest is selected from an enzyme (e.g., a lysosomal enzyme), a nanoparticle, a viral composition (e.g., a viral particle), a therapeutic protein, a therapeutic antibody, and a cytotoxic drug.

[0233] In some embodiments, the moiety of interest is a lysosomal enzyme for delivery to cells for use in enzyme replacement therapy, such as acid α-glucosidase (GAA). Lysosomal enzymes of interest that can be adapted for use in the conjugates of the present disclosure include acid α-glucosidase, acid β-galactosidase-1, acid sphingomyelinase, α-D-mannosidase, α-fucosidase, α-galactosidase A, α-glucosaminide acetyltransferase, α-glucosidase, α-L-iduronidase, α-N-acetylgalactosaminidase, α-acetylglucosaminidase, α-D-neuraminidase, arylsulfatase A, arylsulfatase B, β-galactosidase, β-glucuronidase, β-mannosidase, cathepsin D, cathepsin K, ceramidase, cystinosin, ganglioside activator GM2, These include, but are not limited to, galactocerebrosidase, glucocerebrosidase, heparan sulfatase, hexosaminidase A, hexosaminidase B, hyaluronidase, iduronate-2-sulfatase, LAMP2, lysosomal acid lipase, N-acetylglucosamine-1-phosphotransferase, N-acetylgalactosamine-6-sulfatase, N-acetylglucosamine-1-phosphotransferase, N-acetylglucosamine-6-sulfatase, N-aspartyl-β-glucosaminidase, palmitoylthioesterase-1, acid phosphatase, protective protein / cathepsin A (PPCA), sialin, and tripeptidyl peptidase 1.

[0234] Aspects of the present disclosure include compounds of formula (I), wherein the moiety of interest Y is selected from a small molecule, a dye, a fluorophore, a monosaccharide, a disaccharide, a trisaccharide, and a biomolecule. In some embodiments, Y is a small molecule that specifically binds to a target molecule, such as a target protein.

[0235] In some embodiments of the compounds of the present disclosure, Y is a biomolecule. In some embodiments, the biomolecule is selected from a protein, a polynucleotide, a polysaccharide, a peptide, a glycoprotein, a lipid, an enzyme, an antibody, and an antibody fragment. In some embodiments, Y is a biomolecule that specifically binds to a target molecule, such as a target protein.

[0236] The compounds of the present disclosure may sometimes be referred to as conjugates, for example, when the moiety of interest (Y) is a molecule, such as a biomolecule, in which case the conjugate can be derived from a conjugation or coupling reaction between a chemoselective linking group and a compatible group on the biomolecule. In some embodiments, the biomolecule is conjugated via a naturally occurring group on the biomolecule. In some embodiments, the biomolecule is conjugated via a compatible functional group introduced into the biomolecule prior to chemoselective conjugation. In such cases, the linking moiety between X and Y incorporates a residue (e.g., Z) that is the product of the chemoselective conjugation chemistry.

[0237] Aspects of the present disclosure include compounds of formula (I), wherein the moiety of interest Y is a moiety that specifically binds to a target molecule, such as a target protein. The target protein may be a membrane-bound protein or an extracellular protein. In some embodiments of the compounds of the present disclosure, Y is a biomolecule that specifically binds to the target protein. The present disclosure provides conjugates of certain ASGPR-binding compounds with the conjugate. In some embodiments, the conjugates include a moiety of interest Y that specifically binds to a target protein and can be used in methods for cellular uptake or internalization of the target protein via binding to the cell surface receptor, and ultimately for degradation of the target protein.

[0238] In some embodiments, Y is an aptamer that specifically binds to a target molecule such as a target protein. In some embodiments, Y is a peptide or protein (e.g., a peptidic binding motif, a protein domain, an engineered polypeptide, or a glycoprotein) that specifically binds to a target molecule such as a target protein. In some embodiments, Y is an antibody or antibody fragment that specifically binds to a target molecule such as a target protein. In some embodiments, Y is a polynucleotide or oligonucleotide that specifically binds to a target molecule such as a target protein or a target nucleic acid.

[0239] In some embodiments, one Y biomolecule is conjugated via a linker, L, to a single moiety (X) that specifically binds to a cell surface receptor (e.g., ASGPR). In some embodiments, one Y biomolecule is conjugated to one (Xn-L)-group, where when n=1, the (Xn-L)-group is referred to as monovalent, and when n>1, the (Xn-L)-group is referred to as multivalent (e.g., divalent, trivalent, etc.). In some embodiments of Formula (I) where Y is a biomolecule, it is understood that Y may be conjugated to two or more (Xn-L)-groups, where each (Xn-L)-group may itself be monovalent or multivalent (e.g., divalent, trivalent, etc.). In such cases, the ratio of linked (Xn-L)-groups to biomolecule may be referred to as 2 or more.

[0240] In some embodiments, Y is an antibody or antibody fragment that specifically binds to a target protein, and the compound has formula (III): [ka] (In the formula, n is 1 to 20, m is the average load number from 1 to 80, Each X is a moiety that binds to cell surface ASGPR, each L is a linker, each Z is a residue resulting from the covalent attachment of a chemoselective linking group to a compatible group on an Ab; Ab is an antibody or antibody fragment that specifically binds to the target protein. It is a conjugate of

[0241] In certain embodiments of Formula (III), each X is independently of Formula (Ib) (e.g., as described herein). Optionally, each X is independently selected from the compounds in Table 1. Optionally, each X is independently selected from the following compounds: [ka] is selected from one of the following:

[0242] In certain embodiments of Formula (III), each X is independently selected from the following compounds: [ka] (In the formula, R 5 and R 4 are independently H or a pro-moiety, or R 5 and R 4 are linked in a ring to form a pro moiety, n1 and n2 are each independently an integer of 1 to 6, and Y 4 is an appropriate counterion) In some embodiments, Y 4 is sodium.

[0243] In certain embodiments of Formula (III), n is 1 and X is [ka] is.

[0244] In certain other embodiments of Formula (III), each X is independently of Formula (Ic) (e.g., as described herein). Optionally, each X is independently selected from the compounds of Table 2.

[0245] In certain other embodiments of Formula (III), each X is independently of Formula (Id) (e.g., as described herein). Optionally, each X is independently selected from a compound in Table 3. Optionally, each X is independently selected from a compound in Table 4.

[0246] In certain embodiments of Formula (III), each X is independently selected from the following compounds: [ka] is selected from one of the following:

[0247] In certain embodiments of the conjugate of Formula (III), L is a linker of Formula (II) (eg, as described herein).

[0248] In certain embodiments of the conjugate of formula (III), n is 1 to 6. Optionally, n is 1, such that the antibody is conjugated to a monovalent ASGPR ligand, and the linker is of formula (IIa) (e.g., as described herein). Optionally, n is at least 2, such that the antibody is conjugated to a multivalent ASGPR ligand. Optionally, n is 2. Optionally, n is 3.

[0249] In certain embodiments of the conjugate of Formula (III), Z is a residual moiety resulting from the covalent attachment of a chemoselective linking moiety of Table 8.

[0250] In certain embodiments of the conjugate of Formula (III), Z is the residual moiety resulting from the covalent attachment of the thiol-reactive chemoselective linking group to one or more cysteine ​​residue(s) of the Ab.

[0251] In certain other embodiments of the conjugate of Formula (III), Z is a residual moiety resulting from the covalent attachment of an amine-reactive chemoselective linking group to one or more lysine residue(s) of Ab.

[0252] In certain embodiments, conjugates having the linker structures described herein have weaker binding affinity to cell surface receptors. Without being bound by any particular mechanism or theory, such weaker binding affinity may be associated with a longer half-life of the conjugate and may be useful for adjusting (e.g., modifying) the pharmacokinetic properties of the conjugates described herein. In certain embodiments, even such weakly bound conjugates have sufficiently strong uptake.

[0253] Conjugates of a polypeptide, e.g., an antibody (Ab), and a compound (Xn-LY) can be made using a variety of bifunctional protein coupling agents, such as BMPS, EMCS, GMBS, HBVS, LC-SMCC, MBS, MPBH, SBAP, SIA, SIAB, SMCC, SMPB, SMPH, sulfo-EMCS, sulfo-GMBS, sulfo-KMUS, sulfo-MBS, sulfo-SIAB, sulfo-SMCC, sulfo-SMPB, and SVSB (succinimidyl-(4-vinylsulfone)benzoate). The present disclosure further contemplates that the conjugates described herein may be prepared using any suitable method disclosed in the art (see, e.g., Bioconjugate Techniques (Hermanson ed., 2d ed. 2008)).

[0254] In certain embodiments of the conjugates described herein, L is attached to a lysine residue of the polypeptide via an amide bond. In certain embodiments of the conjugates described herein, L is attached to a cysteine ​​residue of the polypeptide via a thioether bond. In certain embodiments of the conjugates described herein, L is attached to a lysine residue of the Ab via an amide bond. In certain embodiments of the conjugates described herein, L is attached to a cysteine ​​residue of the Ab via a thioether bond. In certain embodiments of the conjugates described herein, L is attached to two cysteine ​​residues of the Ab via two thioether bonds, the two cysteine ​​residues being derived from cleaved cysteine-cysteine ​​disulfide bonds of the Ab. In certain embodiments, the cleaved cysteine-cysteine ​​disulfide bonds are interchain disulfide bonds.

[0255] In certain embodiments of the conjugates described herein, when L is attached to a lysine residue of a polypeptide (e.g., an antibody) via an amide bond, m is an integer from 1 to 80. In certain embodiments of the conjugates described herein, when L is attached to a cysteine ​​residue of P via a thioether bond, m is an integer from 1 to 8.

[0256] In certain embodiments, conjugation to a polypeptide or antibody Ab may be via site-specific conjugation. Site-specific conjugation can, for example, result in uniform loading and minimize conjugate subpopulations with potentially altered antigen binding or pharmacokinetics. In certain embodiments, for example, conjugation can involve engineering cysteine ​​substitutions at positions on the polypeptide or antibody, such as positions on the heavy and / or light chains of an antibody, that provide reactive thiol groups and do not disrupt folding and assembly of the polypeptide or antibody, or that alter binding of the polypeptide or antigen (see, e.g., Junutula et al., J. Immunol. Meth. 2008;332:41-52, and Junutula et al., Nature Biotechnol. 2008;26:925-32, and see also WO2006 / 034488, which is incorporated by reference in its entirety. In another non-limiting approach, selenocysteine ​​is co-translationally inserted into a polypeptide or antibody sequence by recoding the stop codon UGA from a termination to a selenocysteine ​​insertion, allowing site-specific covalent conjugation with the nucleophilic selenol group of selenocysteine ​​in the presence of other natural amino acids (see, e.g., Hofer et al., Proc. Natl. Acad. Sci. USA 2008;105:12451-56, and Hofer et al., Biochemistry 2009;48(50):12047-57). Still other non-limiting techniques that allow for site-specific conjugation to polypeptides or antibodies include, for example, engineering unnatural amino acids, including p-acetylphenylalanine (p-acetyl-Phe), p-azidomethyl-N-phenylalanine (p-azidomethyl-Phe), and azidolysine (azido-Lys), at specific linkage sites, and may further include engineering unique functional tags, for example, LPXTG, LLQGA, sialic acid, and GlcNac, for enzyme-mediated conjugation.See Jackson, Org. Process Res. Dev. 2016;20:852-866, and Tsuchikama and An, Protein Cell 2018;9(1):33-46, the contents of each of which are incorporated by reference in their entirety. See also US2019 / 0060481A1 and US2016 / 0060354A1, the contents of each of which are incorporated by reference in their entirety. All such methodologies are contemplated for use in connection with making the conjugates described herein.

[0257] The loading number of a compound of Formula (I) onto a polypeptide (e.g., an antibody) described herein is represented by "m" in Formula (III) and is the average number of "Xn-L-" or "Xn-" units per conjugate molecule. As used herein, the term "DAR" refers to the average value of "m" or the loading number of a conjugate. The number of "X" moieties (e.g., folate moieties) per each unit of "Xn-L-" or "Xn-" is represented by "n" in Formula (III). As used herein, the term "valency(ies)" refers to the number of "X" moieties ("n") per unit. It will be understood that the loading number or DAR is not necessarily equivalent to the number of "X" moieties per conjugate molecule. For example, if there is one "X" moiety per unit (n=1, valency is "1") and one "Xn-L-" unit (m=1) per conjugate, then there is 1 x 1 = 1 "X" moiety per conjugate. However, if there are two "X" moieties per unit (n=2, valency is "2") and four "Xn-L-" units per conjugate (m=4), then there are 2 x 4 = 8 "X" moieties per conjugate. Thus, for the conjugates described herein, the total number of "X" moieties per conjugate molecule is n x m. As used herein, the term "total valency(ies)" refers to the total number of "X" moieties (n x m; total valency) per conjugate molecule.

[0258] The DAR (loading number) ranges from 1 to 80 units per conjugate. The conjugates provided herein can include a collection of polypeptides, antibodies, or antigen-binding fragments conjugated with a range of units, e.g., 1 to 80. The average number of units per polypeptide or antibody in a conjugate preparation from a conjugation reaction can be characterized by conventional means, such as mass spectrometry. The quantitative distribution of DAR (loading number) with respect to m can also be measured. In some cases, separation, purification, and characterization of homogeneous conjugates with a particular value of m can be achieved by means, such as electrophoresis.

[0259] In certain embodiments, the DAR of the conjugates provided herein ranges from 1 to 80. In certain embodiments, the DAR of the conjugates provided herein ranges from 1 to 70. In certain embodiments, the DAR of the conjugates provided herein ranges from 1 to 60. In certain embodiments, the DAR of the conjugates provided herein ranges from 1 to 50. In certain embodiments, the DAR of the conjugates provided herein ranges from 1 to 40. In certain embodiments, the DAR of the conjugates provided herein ranges from 1 to 35. In certain embodiments, the DAR of the conjugates provided herein ranges from 1 to 30. In certain embodiments, the DAR of the conjugates provided herein ranges from 1 to 25. In certain embodiments, the DAR of the conjugates provided herein ranges from 1 to 20. In certain embodiments, the DAR of the conjugates provided herein ranges from 1 to 18. In certain embodiments, the DAR of the conjugates provided herein ranges from 1 to 15. In certain embodiments, the DAR of the conjugates provided herein ranges from 1 to 12. In certain embodiments, the DAR of the conjugates provided herein ranges from 1 to 10. In certain embodiments, the DAR of the conjugates provided herein ranges from 1 to 9. In certain embodiments, the DAR of the conjugates provided herein ranges from 1 to 8. In certain embodiments, the DAR of the conjugates provided herein ranges from 1 to 7. In certain embodiments, the DAR of the conjugates provided herein ranges from 1 to 6. In certain embodiments, the DAR of the conjugates provided herein ranges from 1 to 5. In certain embodiments, the DAR of the conjugates provided herein ranges from 1 to 4. In certain embodiments, the DAR of the conjugates provided herein ranges from 1 to 3. In certain embodiments, the DAR of the conjugates provided herein ranges from 2 to 12.In certain embodiments, the DAR of the conjugates provided herein ranges from 2 to 10. In certain embodiments, the DAR of the conjugates provided herein ranges from 2 to 9. In certain embodiments, the DAR of the conjugates provided herein ranges from 2 to 8. In certain embodiments, the DAR of the conjugates provided herein ranges from 2 to 7. In certain embodiments, the DAR of the conjugates provided herein ranges from 2 to 6. In certain embodiments, the DAR of the conjugates provided herein ranges from 2 to 5. In certain embodiments, the DAR of the conjugates provided herein ranges from 2 to 4. In certain embodiments, the DAR of the conjugates provided herein ranges from 3 to 12. In certain embodiments, the DAR of the conjugates provided herein ranges from 3 to 10. In certain embodiments, the DAR of the conjugates provided herein ranges from 3 to 9. In certain embodiments, the DAR of the conjugates provided herein ranges from 3 to 8. In certain embodiments, the DAR of the conjugates provided herein ranges from 3 to 7. In certain embodiments, the DAR of the conjugates provided herein ranges from 3 to 6. In certain embodiments, the DAR of the conjugates provided herein ranges from 3 to 5. In certain embodiments, the DAR of the conjugates provided herein ranges from 3 to 4.

[0260] In certain embodiments, the DAR of the conjugates provided herein ranges from 1 to about 8, from about 2 to about 6, from about 3 to about 5, from about 3 to about 4, from about 3.1 to about 3.9, from about 3.2 to about 3.8, from about 3.2 to about 3.7, from about 3.2 to about 3.6, from about 3.3 to about 3.8, or from about 3.3 to about 3.7.

[0261] In certain embodiments, the DAR of the conjugates provided herein is about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, or more. In some embodiments, the DAR of the conjugates provided herein is about 3.1, about 3.2, about 3.3, about 3.4, about 3.5, about 3.6, about 3.7, about 3.8, or about 3.9.

[0262] In some embodiments, the DAR of the conjugates provided herein is in the range of 2 to 20, 2 to 19, 2 to 18, 2 to 17, 2 to 16, 2 to 15, 2 to 14, or 2 to 13. In some embodiments, the DAR of the conjugates provided herein is in the range of 3 to 20, 3 to 19, 3 to 18, 3 to 17, 3 to 16, 3 to 15, 3 to 14, or 3 to 13. In some embodiments, the DAR of the conjugates provided herein is about 1. In some embodiments, the DAR of the conjugates provided herein is about 2. In some embodiments, the DAR of the conjugates provided herein is about 3. In some embodiments, the DAR of the conjugates provided herein is about 4. In some embodiments, the DAR of the conjugates provided herein is about 3.8. In some embodiments, the DAR of the conjugates provided herein is about 5. In some embodiments, the DAR of the conjugates provided herein is about 6. In some embodiments, the DAR of the conjugates provided herein is about 7. In some embodiments, the DAR of the conjugates provided herein is about 8. In some embodiments, the DAR of the conjugates provided herein is about 9. In some embodiments, the DAR of the conjugates provided herein is about 10. In some embodiments, the DAR of the conjugates provided herein is about 11. In some embodiments, the DAR of the conjugates provided herein is about 12. In some embodiments, the DAR of the conjugates provided herein is about 13. In some embodiments, the DAR of the conjugates provided herein is about 14. In some embodiments, the DAR of the conjugates provided herein is about 15. In some embodiments, the DAR of the conjugates provided herein is about 16. In some embodiments, the DAR of the conjugates provided herein is about 17. In some embodiments, the DAR of the conjugates provided herein is about 18.In some embodiments, the DAR of the conjugates provided herein is about 19. In some embodiments, the DAR of the conjugates provided herein is about 20.

[0263] In some embodiments, the DAR of the conjugates provided herein is about 25. In some embodiments, the DAR of the conjugates provided herein is about 30. In some embodiments, the DAR of the conjugates provided herein is about 35. In some embodiments, the DAR of the conjugates provided herein is about 40. In some embodiments, the DAR of the conjugates provided herein is about 50. In some embodiments, the DAR of the conjugates provided herein is about 60. In some embodiments, the DAR of the conjugates provided herein is about 70. In some embodiments, the DAR of the conjugates provided herein is about 80.

[0264] In certain embodiments, fewer than the theoretical maximum number of units are conjugated to a polypeptide, e.g., an antibody, during a conjugation reaction. The polypeptide may contain, for example, lysine residues that do not react with a compound or a linker reagent. Generally, for example, antibodies do not contain many free and reactive cysteine ​​thiol groups that can be linked to a Drug unit; in fact, most cysteine ​​thiol residues of antibodies exist as disulfide bridges. In certain embodiments, antibodies can be reduced with a reducing agent such as dithiothreitol (DTT) or tricarbonylethylphosphine (TCEP) under partial or total reducing conditions to generate reactive cysteine ​​thiol groups. In certain embodiments, the antibody is subjected to denaturing conditions to reveal reactive nucleophilic groups such as lysine or cysteine. In some embodiments, a compound is conjugated via a lysine residue on the antibody. In some embodiments, a linker unit or Drug unit is conjugated via a cysteine ​​residue on the antibody.

[0265] In certain embodiments, the amino acid that binds to the unit is present in the heavy chain of the antibody. In certain embodiments, the amino acid that binds to the unit is present in the light chain of the antibody. In certain embodiments, the amino acid that binds to the unit is present in the hinge region of the antibody. In certain embodiments, the amino acid that binds to the unit is present in the Fc region of the antibody. In certain embodiments, the amino acid that binds to the unit is present in the constant region of the antibody (e.g., CH1, CH2, or CH3 of the heavy chain, or CH1 of the light chain). In yet other embodiments, the amino acid that binds to the unit or Drug Unit is present in the VH framework region of the antibody. In yet other embodiments, the amino acid that binds to the unit is present in the VL framework region of the antibody.

[0266] The DAR (loading number) of the conjugate can be controlled in different ways, for example, by (i) limiting the molar excess of the compound or conjugation reagent compared to the polypeptide, (ii) limiting the conjugation reaction time or temperature, (iii) partial or limited reducing conditions for cysteine ​​thiol modifications, or (iv) manipulating the amino acid sequence of the polypeptide by recombinant techniques such that the number and position of cysteine ​​residues are altered to adjust the number and / or position of linker-drug bonds (such as thiomab prepared as disclosed in WO2006 / 034488, which is incorporated herein by reference in its entirety).

[0267] It should be understood that the preparation of the conjugates described herein results in a mixture of conjugates having a distribution of one or more units bound to a polypeptide, for example, an antibody.The individual conjugate molecules in the mixture can be identified by mass spectrometry and separated by HPLC, for example, hydrophobic interaction chromatography (including methods known in the art).In certain embodiments, a homogeneous conjugate having a single DAR (loading number) value can be isolated from the conjugation mixture by electrophoresis or chromatography.

[0268] In certain embodiments of the conjugate of Formula (III), m is 1 to 20, e.g., 2 to 10, 2 to 8, or 2 to 6. Optionally, m is 10 or less. Optionally, m is 2 to 8. Optionally, m is 2 to 6. Optionally, m is an average loading number of about 4.

[0269] It should be understood that the preparation of the conjugate described herein can result in a mixture of conjugates having a distribution of one or more units bound to a polypeptide, for example, an antibody.The individual conjugate molecules in the mixture can be identified by mass spectrometry and separated by HPLC, for example, hydrophobic interaction chromatography (including methods known in the art).In certain embodiments, a homogeneous conjugate having a single DAR (loading number) value can be isolated from the conjugation mixture by electrophoresis or chromatography.

[0270] 5.4.1.Target binding moiety The target binding moiety can be any moiety that has an affinity for the target of less than 1 μM, e.g., 300 nM or less, 100 nM or less, 30 nM or less, 10 nM or less, 3 nM or less, or 1 nM or less, as measured, for example, in an in vitro binding assay.

[0271] In some embodiments, the target binding moiety is a biomolecule. In some embodiments, the target binding moiety is a biomolecule that specifically binds to a target protein. In some embodiments, the biomolecule is selected from a peptide, a protein, a polynucleotide, a polysaccharide, a glycan, a glycoprotein, a lipid, an enzyme, an antibody, and an antibody fragment.

[0272] In some embodiments, the target-binding moiety is a polypeptide (e.g., a peptide or protein-binding motif, a protein domain, a modified polypeptide, or a glycoprotein) that specifically binds to a target molecule, such as a target protein. In some embodiments, the target-binding moiety of the bifunctional compound comprises a polypeptide that binds to a soluble (e.g., secreted) target protein of interest. In some embodiments, the target binding moiety is a polypeptide ligand that comprises a receptor ligand, or a receptor-binding portion or fragment of a receptor ligand, that binds to a target cell surface receptor. Target-binding polypeptides may contain L-amino acids, D-amino acids, or both, and may contain any of a variety of natural amino acids, unnatural amino acids, and / or amino acid modifications or analogs known in the art. Useful modifications include, for example, N-terminal acetylation, amidation, methylation, etc.

[0273] In some embodiments, the target binding moiety is a polynucleotide that specifically binds to a target molecule, such as a target protein or a target nucleic acid. The terms polynucleotide and nucleic acid can be used interchangeably. In some embodiments, the target binding moiety is a nucleic acid aptamer that specifically binds to a target molecule, such as a target protein.

[0274] In some embodiments, the target binding moiety is a glycan. In some embodiments, the target binding moiety is a glycan epitope of an autoantibody.

[0275] 5.4.1.1. Antibodies In some embodiments, the target binding moiety is an antibody or antibody fragment that specifically binds to a target moiety, such as a target protein.

[0276] The ASGPR-binding moiety may be site-specifically covalently linked to the antibody or antibody fragment via an optional linking moiety. The ASGPR-binding moiety may be covalently linked to the antibody or antibody fragment via a site-specific cysteine ​​modification (e.g., L443C) on the antibody or antibody fragment and a thiol-reactive chemoselective linking group. The ASGPR-binding moiety may be covalently linked to the antibody or antibody fragment via one or more lysine residues on the antibody or antibody fragment and an amine-reactive chemoselective linking group.

[0277] In some embodiments, the bifunctional conjugate of the present disclosure comprises an antibody (Ab). In some embodiments, the Ab is a monoclonal antibody. In some embodiments, the Ab is a human antibody. In some embodiments, the Ab is a humanized antibody. In some embodiments, the Ab is a chimeric antibody. In some embodiments, the Ab is a full-length antibody comprising two heavy chains and two light chains. In some embodiments, the Ab is an IgG antibody, e.g., an IgG1, IgG2, IgG3, or IgG4 antibody. In some embodiments, the Ab is a single-chain antibody. In some embodiments, the target-binding moiety is an antigen-binding fragment of an antibody, e.g., a Fab fragment.

[0278] In some embodiments, the antibody or antibody fragment specifically binds to a cancer antigen.

[0279] In some embodiments, the antibody or antibody fragment specifically binds to a hepatocyte antigen.

[0280] In some embodiments, the antibody or antibody fragment specifically binds to an antigen presented on a macrophage.

[0281] In some embodiments, the antibody or antibody fragment specifically binds to intact complement or a fragment thereof, hi some embodiments, the antibody or antibody fragment specifically binds to one or more immunodominant epitope(s) within intact complement or a fragment thereof.

[0282] In some embodiments, the antibody or antibody fragment specifically binds to a cell surface receptor. In some embodiments, the antibody or antibody fragment specifically binds to a cell surface receptor ligand.

[0283] In some embodiments, the antibody or antibody fragment specifically binds to an epidermal growth factor (EGF) protein, e.g., human EGF, hi some embodiments, the antibody or antibody fragment specifically binds to one or more immunodominant epitopes within the EGF protein.

[0284] In some embodiments, the antibody or antibody fragment specifically binds to an epidermal growth factor receptor (EGFR) protein, e.g., human EGFR. In some embodiments, the antibody or antibody fragment specifically binds to one or more immunodominant epitopes within the EGFR protein. In some embodiments, the antibody or antibody fragment comprises CDRs present in cetuximab. In some embodiments, the antibody or antibody fragment comprises a variable light chain and a variable heavy chain present in cetuximab. In some embodiments, the antibody is cetuximab. In some embodiments, the antibody or antibody fragment comprises CDRs present in matuzumab. In some embodiments, the antibody or antibody fragment comprises a variable light chain and a variable heavy chain present in matuzumab. In some embodiments, the antibody is matuzumab.

[0285] In some embodiments, the antibody or antibody fragment specifically binds to a vascular endothelial growth factor (VEGF) protein, such as a human VEGF protein, hi some embodiments, the antibody or antibody fragment specifically binds to one or more immunodominant epitopes within the VEGF protein.

[0286] In some embodiments, the antibody or antibody fragment specifically binds to a vascular endothelial growth factor receptor (VEGFR) protein, such as a human VEGFR protein. In some embodiments, the antibody or antibody fragment specifically binds to a vascular endothelial growth factor receptor 2 (VEGFR2) protein, such as a human VEGFR2 protein. In some embodiments, the antibody or antibody fragment specifically binds to a vascular endothelial growth factor receptor 3 (VEGFR3) protein, such as a human VEGFR3 protein. In some embodiments, the antibody or antibody fragment specifically binds to one or more immunodominant epitopes within a VEGFR protein, a VEGFR2 protein, or a VEGFR3 protein.

[0287] In some embodiments, the antibody or antibody fragment specifically binds to fibroblast growth factor (FGF), e.g., human FGF, hi some embodiments, the antibody or antibody fragment specifically binds to one or more immunodominant epitopes within the FGF protein.

[0288] In some embodiments, the antibody or antibody fragment specifically binds to a fibroblast growth factor receptor (FGFR), such as human FGFR. In some embodiments, the antibody or antibody fragment specifically binds to a fibroblast growth factor receptor 2 (FGFR2) protein, such as human FGFR2 protein, such as FGFR2b protein. In some embodiments, the antibody or antibody fragment specifically binds to a fibroblast growth factor receptor 3 (FGFR3) protein, such as human FGFR3 protein. In some embodiments, the antibody or antibody fragment specifically binds to one or more immunodominant epitopes within an FGFR protein, FGFR2 protein, or FGFR3 protein.

[0289] In some embodiments, the antibody specifically binds to the receptor tyrosine kinase cMET protein. In some embodiments, the antibody specifically binds to one or more immunodominant epitope(s) within the receptor tyrosine kinase cMET protein.

[0290] In some embodiments, the antibody specifically binds to a CD47 protein, e.g., a human CD47 protein, hi some embodiments, the antibody specifically binds to one or more immunodominant epitope(s) within the CD47 protein.

[0291] In some embodiments, the antibody specifically binds to an immune checkpoint inhibitor. In some embodiments, the antibody binds to one or more immunodominant epitopes within the immune checkpoint inhibitor. In some embodiments, the antibody specifically binds to a programmed cell death protein, such as human PD-1. In some embodiments, the antibody specifically binds to one or more immunodominant epitopes within the PD-1 protein.

[0292] In some embodiments, the antibody specifically binds to a programmed death-ligand-1 (PD-L1) protein, such as human PD-L1, hi some embodiments, the antibody specifically binds to one or more immunodominant epitope(s) within the PD-L1 protein.

[0293] In some embodiments, the antibody binds to TIM3, hi some embodiments, the antibody binds to one or more immunodominant epitope(s) within TIM3.

[0294] In some embodiments, the antibody specifically binds to a lectin. In some embodiments, the antibody specifically binds to one or more immunodominant epitopes within a lectin. In some embodiments, the antibody binds to SIGLEC. In some embodiments, the antibody binds to one or more immunodominant epitopes within SIGLEC. In some embodiments, the antibody binds to a cytokine receptor. In some embodiments, the antibody binds to one or more immunodominant epitopes within a cytokine receptor. In some embodiments, the antibody binds to sIL6R. In some embodiments, the antibody binds to one or more immunodominant epitopes within sIL6R. In some embodiments, the antibody binds to a cytokine. In some embodiments, the antibody binds to one or more immunodominant epitopes within a cytokine. In some embodiments, the antibody binds to MCP-1, TNF (e.g., TNF-α), IL1a, IL1b, IL4, IL5, IL6, IL12 / IL23, IL13, IL17, or p40. In some embodiments, the antibody binds to one or more immunodominant epitope(s) within MCP-1, TNF (e.g., TNF-α), IL1a, IL1b, IL4, IL5, IL6, IL12 / IL23, IL13, IL17, or p40.

[0295] In some embodiments, the antibody binds to a major histocompatibility protein (e.g., an MHC class I or class II molecule). In some embodiments, the antibody binds to one or more immunodominant epitope(s) within a major histocompatibility protein (e.g., an MHC class I or class II molecule). In some embodiments, the antibody binds to β2 microglobulin. In some embodiments, the antibody binds to one or more immunodominant epitope(s) within β2 microglobulin.

[0296] 5.4.1.2. Modified Virus Compositions In certain embodiments, Y is a viral particle, a viral capsid, a viral envelope, or a viral protein. In some embodiments, the viral composition is a viral particle comprising a transgene. In some embodiments, the viral protein is a viral capsid protein or a viral envelope protein.

[0297] In certain aspects, provided herein are modified viral compositions comprising a viral composition, e.g., a viral particle, a viral capsid, or a viral protein (e.g., a viral capsid protein or an envelope protein), linked (e.g., directly or indirectly, e.g., via an intervening linker sequence) to an ASGPR-binding moiety that binds to a cell surface receptor. In certain embodiments, the modified viral composition comprises a viral particle that optionally includes a polynucleotide comprising a transgene, e.g., a transgene useful for therapeutic applications.

[0298] The modified viral compositions, e.g., viral conjugates, provided herein can include any viral composition described herein, e.g., any viral particle, capsid, or viral protein, e.g., capsid protein or envelope protein, or fragment thereof, described herein.

[0299] In certain embodiments, the viral compositions described herein may comprise viral particles. The terms "virus particle," "viral particle," "virus vector," or "viral vector" are used interchangeably herein. A "viral particle" refers to a viral capsid and a polynucleotide (DNA or RNA), which may comprise the viral genome, a portion of the viral genome, or a polynucleotide derived from the viral genome (e.g., one or more ITRs), which optionally comprises a transgene. In certain cases, a viral particle further comprises an envelope (generally comprising a lipid portion and envelope proteins) surrounding or partially surrounding the capsid.

[0300] Viral particles may also be referred to as "recombinant viral particles" or "recombinant virus particles," as that term is used herein to refer to viral particles that have been genetically altered, for example, by deletion or other mutation of endogenous viral genes and / or the addition or insertion of a heterologous nucleic acid construct into the polynucleotide of the viral particle. Thus, a recombinant viral particle generally refers to a viral particle that includes a capsid coat or shell (and optional outer envelope) within which is encapsulated a polynucleotide sequence that includes sequences of viral and non-viral origin (i.e., a polynucleotide heterologous to the virus). This polynucleotide sequence is generally the sequence that is the subject of cellular genetic alteration.

[0301] In certain aspects, the viral compositions described herein, when referred to herein in the context of viruses, may comprise a "viral capsid," "empty viral particle," "empty virus particle," or "capsid," or "empty particle," which terms as used herein refer to a three-dimensional shell or coat comprising viral capsid proteins, optionally surrounded or partially surrounded by an outer envelope. In certain embodiments, the viral composition is a viral particle or fragment thereof, a viral capsid or fragment thereof, a viral protein, e.g., a viral capsid protein or fragment thereof, or an envelope protein or fragment thereof.

[0302] In some embodiments, the viruses used in the modified virus compositions provided herein are adenoviruses (AV); adeno-associated viruses (AAV); retroviruses (e.g., lentiviruses (LV), rhabdoviruses, murine leukemia viruses); herpes simplex viruses, coronaviruses, reoviruses, etc. In some embodiments, the viral vectors, viral particles, or viral proteins used in the present disclosure are derived from non-enveloped viruses, such as adeno-associated viruses (AAV).

[0303] In some embodiments, lentiviral vectors can be used for CAR-T gene delivery, vaccines, or research tools (e.g., to introduce genes into mature T cells to elicit immunity against cancer through delivery of chimeric antigen receptors (CARs) or cloned T cell receptors).

[0304] Naturally occurring AAV forms viral particles that include a three-dimensional capsid coat or shell ("capsid") composed of capsid proteins (VP1, VP2, and VP3) and the AAV viral genome contained within the capsid.

[0305] The modified AAV compositions provided herein, e.g., AAV conjugates or fusions, may comprise any AAV composition described herein, e.g., any AAV particle, capsid, or capsid protein described herein, or a fragment thereof. The term "AAV capsid protein" or "AAV cap protein" refers to a protein encoded by an AAV capsid gene (e.g., VP1, VP2, and VP3) or a variant or fragment thereof. The term includes capsid proteins expressed by or derived from AAV, e.g., recombinant AAV, such as chimeric AAV. For example, the term includes, but is not limited to, capsid proteins from any AAV serotype, such as AAV1, AAV2, AAV2i8, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAVrhlO, AAV11, AAV12, AAV13, AAV-DJ, AAV3b, AAVLK03, AAVrh74, AAVAnc81, Anc82, Anc83, Anc84, AncllO, Ancll3, Anc126, or Anc127, AAV_go.1, AAVhu.37, or AAVrh.8, or variants thereof.

[0306] 5.4.1.3. Bridging Moieties Binding to Viral Compositions In some embodiments, Y is a bridging moiety that specifically binds to the viral composition, e.g., a viral particle, a viral capsid, a viral envelope, or a viral protein (e.g., a viral capsid protein or envelope protein), provided that the binding is not via a covalent bond.

[0307] Any suitable moiety that binds to a viral particle, viral capsid, viral envelope, or viral protein (e.g., a viral capsid protein or envelope protein) may be adapted for use in the conjugates of the present disclosure.

[0308] In certain embodiments, the bridging moiety is a polypeptide that specifically binds to a viral composition. In some embodiments, the bridging moiety is a polypeptide that binds to a viral composition, such as a viral particle, a viral capsid, a viral envelope, or a viral protein, such as a viral capsid protein or a viral envelope protein. In certain aspects, when the viral protein is part of a viral particle, the bridging composition binds to a viral capsid protein or a viral envelope protein.

[0309] In certain embodiments, the bridging moiety is an antibody or antibody fragment (e.g., an antigen-binding fragment of an antibody) that specifically binds to the viral composition. In certain embodiments, a bridging moiety that binds to a viral protein may also bind to a viral particle, for example, via binding to the viral protein incorporated into the viral particle. Similarly, in certain embodiments, a bridging moiety that binds to a viral particle may also bind to the viral protein even if the viral protein is not incorporated into the viral particle. The viral particle may be an AAV viral particle. The viral protein may be an AAV capsid protein.

[0310] In some embodiments, a cross-linking moiety of the present disclosure specifically binds to an AAV composition, e.g., an AAV particle, an AAV capsid, or an AAV viral protein (e.g., an AAV capsid protein, e.g., a VP1, VP2, or VP3 protein).

[0311] Antibodies or antigen-binding fragments that can be utilized in connection with the modified virus compositions provided herein, for example, in connection with the crosslinking compositions and crosslinking moieties provided herein, include, but are not limited to, monoclonal antibodies, antibody compositions with specificity for multiple epitopes or for a single epitope, polyclonal or monovalent antibodies, multivalent antibodies, multispecific antibodies (e.g., bispecific antibodies, so long as they exhibit the desired biological activity) formed from at least two intact antibodies, single-chain antibodies, and fragments thereof (e.g., domain antibodies).

[0312] 5.4.1.4.Small molecules In some embodiments, the target-binding moiety of the bifunctional compound of the present disclosure is a small molecule that specifically binds to a target molecule, such as a target protein. In some embodiments, the bifunctional compound includes a small molecule inhibitor or a ligand of the target protein. The small molecule target-binding moiety may be covalently linked to one or more ASGPR-binding moieties via a linker. The linker may be covalently linked to the small molecule via substitution at any suitable site on the small molecule such that binding to the target protein is substantially maintained.

[0313] In some embodiments, the target binding moiety is a small molecule inhibitor or antagonist of the target protein (e.g., as described herein). Any convenient small molecule known to bind to a target of interest may be adapted for use in the subject compounds and conjugates.

[0314] In some embodiments, the target binding moiety is a small molecule inhibitor or antagonist of VEGF.

[0315] In some embodiments, the target-binding moiety is a small molecule inhibitor or antagonist of PD-L1.

[0316] In some embodiments, the target binding moiety is a small molecule inhibitor or antagonist of an EGFR protein, a VEGFR protein, an FGFR2 protein, or an FGFR3 protein.

[0317] In some embodiments, the target binding moiety is a small molecule inhibitor or antagonist of a TNF protein (e.g., TNF-alpha). TNF-alpha (TNFα) is a soluble cytokine produced by monocytes and macrophages as part of immune and inflammatory processes and is involved in a diverse range of cellular responses, including differentiation, proliferation, inflammation, and cell death. TNFα is a type II transmembrane protein that may be cleaved and secreted in a soluble form. Both the transmembrane and soluble biologically active forms of TNFα are homotrimeric complexes that can signal through TNF receptors 1 and 2 (TNF-R1 and TNF-R2). TNFα is directly involved in systemic inflammation through regulation of intracellular NF-κB, JNK, and p38-MAPK signaling pathways.

[0318] The TNFα binding moiety can be a TNFα inhibitor, such as a competitive inhibitor of TNF receptor binding or an allosteric inhibitor of TNF signaling. Compounds of the present disclosure can include potent TNFα inhibitors, e.g., inhibitors with submicromolar inhibitory activity. In some embodiments, the TNFα inhibitor is an allosteric inhibitor. In some embodiments, the TNFα binding moiety is an allosteric desymmetrizing TNFα inhibitor. An allosteric desymmetrizing TNFα inhibitor refers to a compound that binds to an allosteric site in TNFα and stabilizes the trimer unit in an asymmetric conformation that causes the TNFα trimer to recruit only two of the three copies of a TNF receptor (TNFR, e.g., TNFR1), resulting in an incompetent TNFα-TNFR signaling complex.

[0319] See, for example, Xiao et al. in Journal of Medicinal Chemistry 2020 63(23), 15050-15071 and McMillan et al. in Nature Communications (2021) 12:582, which discloses an analysis of the X-ray co-crystal structure of exemplary inhibitors bound to TNFα. Allosteric desymmetrized TNFα inhibitors act via specific mechanisms of action and can exhibit potent inhibitory activity. For example, (a) the binding site of TNFα inhibitors is a cavity within the TNFα trimer created by the displacement of monomer A, (b) the inhibitor stabilizes the TNFα trimer in an inactive conformation by forming critical π-π and hydrogen-bonding interactions, (c) allosteric desymmetrizing TNFα inhibitors bind to the TNFα trimer, largely disrupting one TNFR-binding site and slightly disrupting a second site, while leaving the third site intact, and (d) allosteric desymmetrizing TNFα inhibitors regulate TNF-R activity through an allosteric mechanism rather than direct competition with TNFR. Thus, binding of allosteric desymmetrizing TNFα inhibitors to a symmetric TNFα trimer can result in the formation of an asymmetric trimer that prevents the recruitment of the three TNF receptor molecules required for signal transduction.

[0320] 5.4.2.Target As summarized above, the bifunctional compounds of the present disclosure may include a moiety of interest (Y) that specifically binds to a target molecule, which may be a cell surface molecule or an extracellular molecule.

[0321] In some embodiments of the compounds and methods of the present disclosure, the target molecule is a cell surface molecule. By "cell surface molecule" is meant a target molecule that is associated with a cell membrane, for example, because the molecule has a domain that is inserted into or spans the cell membrane, such as a cell membrane anchoring domain or a transmembrane domain. The cell surface molecule may be any cell surface molecule for which targeted degradation via the endosomal / lysosomal pathway is desirable. In some embodiments, the cell surface molecule is a cell surface receptor.

[0322] Cell surface receptors of interest include, but are not limited to, stem cell receptors, immune cell receptors, growth factor receptors, cytokine receptors, hormone receptors, receptor tyrosine kinases, receptors of the epidermal growth factor receptor (EGFR) family (e.g., HER2 (human epidermal growth factor receptor 2)), receptors of the fibroblast growth factor receptor (FGFR) family, receptors of the vascular endothelial growth factor receptor (VEGFR) family, receptors of the platelet-derived growth factor receptor (PDGFR) family, receptors of the rearranged during transfection (RET) receptor family, receptors of the Eph receptor family, receptors of the discoidin domain receptor (DDR) family, and mucin proteins (e.g., MUC1). In some embodiments, the cell surface molecule is CD71 (transferrin receptor). In certain aspects, the cell surface receptor is an immune cell receptor selected from T cell receptors, B cell receptors, natural killer (NK) cell receptors, macrophage receptors, monocyte receptors, neutrophil receptors, dendritic cell receptors, mast cell receptors, basophil receptors, and eosinophil receptors.

[0323] In some embodiments, the moiety of interest (Y) specifically binds to a cell surface molecule that mediates its effect through bulk biophysical or aggregation effects rather than through specific molecular interactions (and therefore less susceptible to blocking). Non-limiting examples of such cell surface molecules are mucins. Examples of mucins include, but are not limited to, MUC1, MUC16, MUC2, MUC5AC, MUC4, CD43, CD45, GPIb, etc.

[0324] In some embodiments, when the moiety of interest specifically binds to a cell surface molecule, the cell surface molecule is present on a cancer cell. "Cancer cell" refers to a cell that exhibits a neoplastic cell phenotype, which can be characterized by, for example, one or more of aberrant cell growth, aberrant cellular proliferation, loss of density-dependent growth inhibition, anchorage-independent growth potential, the ability to promote tumor growth and / or development in immunocompromised non-human animal models, and / or any suitable indicator of cellular transformation. "Cancer cell" may be used interchangeably herein with "tumor cell," "malignant cell," or "cancerous cell" and includes cancer cells such as solid tumors, semi-solid tumors, hematological malignancies (e.g., leukemia cells, lymphoma cells, myeloma cells, etc.), primary tumors, and metastatic tumors. In some embodiments, the cell surface molecule present on the cancer cell is a tumor-associated antigen or a tumor-specific antigen. In certain aspects, when the moiety of interest (Y) specifically binds to a cell surface molecule, the cell surface molecule is present on an immune cell. In some embodiments, the cell surface molecule is present on an immune cell selected from T cells, B cells, natural killer (NK) cells, macrophages, monocytes, neutrophils, dendritic cells, mast cells, basophils, and eosinophils. In certain aspects, the cell surface molecule present on the immune cell is an inhibitory immunoreceptor. As used herein, an "inhibitory immunoreceptor" is a receptor present on an immune cell that negatively regulates an immune response. Examples of inhibitory immunoreceptors that may be inhibited according to the methods of the present disclosure include inhibitory immunoreceptors of the Ig superfamily, including, but not limited to, CD200R, CD300a (IRp60; mouse MAIR-I), CD300f (IREM-1), CEACAM1 (CD66a), FcyRIIb, ILT-2 (LIR-1; LILRB1; CD85j), ILT-3 (LIR-5; CD85k; LILRB4), ILT-4 (LIR-2; LILRB2), ILT-5 (LIR-3; LILRB3; mouse PIR-B); LAIR-1, PECAM-1 (CD31), PILR-a (FDF03), SIRL-1, and SIRP-a.Further examples of inhibitory immunoreceptors that may be inhibited according to the methods of the present disclosure include sialic acid-binding Ig-like lectin (Siglec) receptors, such as Siglec 7 and Siglec 9. Further examples of inhibitory immunoreceptors that may be inhibited according to the methods of the present disclosure include C-type lectins, including, but not limited to, CLEC4A (DCIR), Ly49Q, and MICL. More information regarding inhibitory immunoreceptors can be found, for example, in Steevels et al. (2011) Eur. J. Immunol. 41(3):575-587. In some embodiments, the cell surface molecule present on the immune cell is a ligand of an inhibitory immunoreceptor. In certain aspects, the cell surface molecule present on the immune cell is an immune checkpoint molecule. Non-limiting examples of immune checkpoint molecules to which the moiety of interest (Y) may specifically bind include PD-1, PD-L1, CTLA4, TIM3, LAG3, TIGIT, and members of the B7 family.

[0325] In some embodiments of the compounds and methods of the present disclosure, the target molecule is an extracellular molecule. By "extracellular molecule" is meant a soluble molecule that is located outside the cell membrane of any cell in the vicinity of the soluble molecule. The extracellular molecule may be any extracellular molecule for which targeted degradation via the endosomal / lysosomal pathway is desirable.

[0326] In some embodiments, the extracellular molecule is a soluble target protein. In some embodiments, the extracellular molecule is a secreted protein that accumulates in disease (e.g., alpha-synuclein), a cholesterol carrier (e.g., ApoB), an infectious disease toxin (e.g., AB toxin, ESAT-6), an infectious particle (e.g., whole virus, whole bacteria, etc.), a coagulation factor (e.g., factor IX), a target of an FDA-approved antibody that binds to the extracellular molecule (e.g., TNFα), any chemokine or cytokine (e.g., mediator of sepsis or chronic inflammation such as IL-1), a protein hormone (e.g., insulin, ACTH, etc.), a protein mediator of mood disorder, a protein mediator of energy homeostasis (e.g., leptin, ghrelin, etc.), a protein allergen present in the bloodstream or an antibody against such an allergen (e.g., for peanut allergy), a protein toxin (e.g., snake venom hyaluronidase, etc.), an autoantibody, etc.

[0327] In some embodiments, the target molecule is an extracellular molecule that is an antibody, e.g., an antibody that specifically binds to a cell surface molecule or a different extracellular molecule. In some embodiments, the antibody is an autoantibody. In some embodiments, the target is human immunoglobulin A (IgA). In some embodiments, the IgA is a specific antibody that plays an important role in mucosal immune function. In the blood, IgA interacts with an Fc receptor called CD89 expressed on immune effector cells to initiate inflammatory responses. Abnormal IgA expression is involved in many autoimmune and immune-mediated diseases. In some embodiments, the target is human immunoglobulin G (IgG). The Fc region of IgG contains a conserved N-glycosylation site at asparagine 297 in the constant region of the heavy chain. Various N-glycans can bind to this site. N-glycan IgG composition has been associated with several autoimmune, infectious, and metabolic diseases. Furthermore, overexpression of IgG4 has been associated with IG4-associated diseases. In some embodiments, the target is human immunoglobulin E (IgE). IgE is a type of immunoglobulin that plays an important role in type I hypersensitivity, which can manifest in a variety of allergic disorders and diseases.

[0328] In some embodiments, the extracellular molecule is a ligand for a cell surface receptor. Cell surface receptor ligands of interest include, but are not limited to, growth factors (e.g., epidermal growth factor (EGF), vascular endothelial growth factor (VEGF), etc.), cytokines (e.g., interleukins, interferons, tumor necrosis factor (TNF), transforming growth factor b (TGF-b) (including certain subtypes of such cytokines)), hormones, etc. In certain embodiments, the moiety of interest (Y) specifically binds to apolipoprotein E4 (ApoE4).

[0329] Pharmaceutical Compositions In another embodiment, provided herein is a pharmaceutical composition comprising one or more conjugates disclosed herein and a pharmaceutically acceptable carrier.

[0330] In certain embodiments, the pharmaceutical compositions provided herein comprise a therapeutically effective amount of one or more of the conjugates provided herein, and optionally one or more additional prophylactic or therapeutic agents, in a pharmaceutically acceptable carrier.

[0331] Pharmaceutical carriers suitable for administration of the conjugates provided herein include any such carriers known to those skilled in the art to be suitable for the particular mode of administration.

[0332] The conjugates described herein may be formulated as the sole pharmaceutically active ingredient in the composition or may be combined with other active ingredients.

[0333] In certain embodiments, the conjugates are formulated into one or more suitable pharmaceutical formulations, such as solutions, suspensions, powders, sustained-release formulations, or elixirs in sterile solutions or suspensions for parenteral administration, or as transdermal patches and dry powder inhalants.

[0334] In the compositions provided herein, the conjugates described herein may be mixed with a suitable pharmaceutical carrier, and the concentration of the conjugate in the composition may be, for example, effective to deliver an amount upon administration that treats, prevents, or ameliorates a disease or disorder described herein, or a symptom thereof.

[0335] In certain embodiments, the pharmaceutical compositions provided herein are formulated for single-dose administration. To formulate the compositions, a weight fraction of the conjugate is dissolved, suspended, dispersed, or otherwise mixed in a selected carrier at an effective concentration such that the disease being treated is alleviated, prevented, or one or more symptoms are ameliorated.

[0336] The concentration of the conjugate in the pharmaceutical compositions provided herein will depend, for example, on the physicochemical properties of the conjugate, the administration schedule and amount administered, as well as other factors known to those of skill in the art.

[0337] The pharmaceutical compositions described herein are provided for administration to a subject, e.g., a human or an animal (e.g., a mammal), in unit dosage forms, such as sterile parenteral (e.g., intravenous) solutions or suspensions containing a suitable amount of the compound or a pharmaceutically acceptable derivative thereof. Pharmaceutical compositions are also provided for administration to humans and animals in unit dosage forms, including oral or nasal solutions or suspensions and oil-in-water emulsions containing an appropriate amount of the conjugate or a pharmaceutically acceptable derivative thereof. In certain embodiments, the conjugate is formulated and administered in unit dosage or multiple dosage forms. As used herein, unit dosage form refers to an individually packaged, physically discrete unit suitable for human or animal (e.g., mammalian) subjects, as known in the art. Each unit dose contains a predetermined amount of the conjugate sufficient to produce the desired therapeutic effect, together with the necessary pharmaceutical carrier, vehicle, or diluent. Examples of unit dosage forms include ampoules and syringes, and individually packaged capsules. Unit dosage forms may be administered in fractions or multiples thereof. A multiple-dose form is a plurality of identical unit-dosage forms packaged in a single container, and administered in separate unit-dosage form.Examples of multiple-dose forms include vials, bottles of capsules, or bottles.Therefore, in certain embodiments, a multiple-dose form is a plurality of unit doses that are not separated by packaging.

[0338] In certain embodiments, the conjugates herein are in the form of liquid pharmaceutical formulations. Liquid pharmaceutically administrable formulations can be prepared, for example, by dissolving, dispersing, or otherwise mixing the conjugate and any pharmaceutical adjuvants in a carrier, such as water, saline, aqueous dextrose, glycerol, glycols, etc., to form a solution or suspension. In certain embodiments, the pharmaceutical compositions provided herein to be administered may also contain small amounts of non-toxic auxiliary substances, such as wetting agents, emulsifying agents, solubilizing agents, and pH buffering agents.

[0339] Actual methods for preparing such dosage forms are known or will be apparent to those skilled in the art; see, for example, Remington: The Science and Practice of Pharmacy (2012) 22nd ed., Pharmaceutical Press, Philadelphia, Pa. Dosage forms or compositions containing between 0.005% and 100% antibody can be made, with the remainder consisting of a non-toxic carrier.

[0340] Parenteral administration, in certain embodiments, is characterized by subcutaneous, intramuscular, or intravenous injection, and is also contemplated herein. Injectables can be prepared in conventional forms as liquid solutions or suspensions, in solid forms suitable for solution or suspension in liquid before injection, or as emulsions. Injectables, solutions, and emulsions also contain one or more excipients. Suitable excipients are, for example, water, saline, dextrose, glycerol, or ethanol. Other routes of administration may include enteral administration, intracerebral administration, intranasal administration, intraarterial administration, intracardiac administration, intraosseous injection, intrathecal administration, and intraperitoneal administration.

[0341] Preparations for parenteral administration include sterile solutions ready for injection, sterile dry soluble products, such as lyophilized powders ready to be mixed with a solvent immediately before use, such as subcutaneous tablets, sterile suspensions ready for injection, sterile dry insoluble products ready to be mixed with a vehicle immediately before use, and sterile emulsions. The solutions may be either aqueous or non-aqueous.

[0342] If administered intravenously, suitable carriers include saline or phosphate buffered saline (PBS), as well as solutions containing thickening and solubilizing agents, such as glucose, polyethylene glycol, and polypropylene glycol, and mixtures thereof.

[0343] Pharmaceutically acceptable carriers used in parenteral formulations include aqueous vehicles, non-aqueous vehicles, antibacterial agents, isotonic agents, buffers, antioxidants, local anesthetics, suspending and dispersing agents, emulsifying agents, sequestering or chelating agents, and other pharmaceutically acceptable substances.

[0344] Pharmaceutical carriers also include ethyl alcohol, polyethylene glycol, and propylene glycol for water miscible vehicles, and sodium hydroxide, hydrochloric acid, citric acid, or lactic acid for pH adjustment.

[0345] In certain embodiments, intravenous or intraarterial infusion of a sterile aqueous solution containing a conjugate described herein is an effective mode of administration. Another embodiment is a sterile aqueous or oily solution or suspension containing a conjugate described herein, injected as needed to produce the desired pharmacological effect.

[0346] In certain embodiments, the pharmaceutical formulations are lyophilized powders, which can be reconstituted for administration as solutions, emulsions, and other mixtures. They may also be reconstituted and formulated as solids or gels.

[0347] The lyophilized powder is prepared by dissolving the conjugate provided herein in a suitable solvent. In some embodiments, the lyophilized powder is sterile. The suitable solvent may contain an excipient that improves the stability or other pharmacological properties of the powder or a reconstituted solution prepared from the powder. Excipients that can be used include, but are not limited to, dextrose, sorbital, fructose, corn syrup, xylitol, glycerin, glucose, sucrose, or other suitable agents. The suitable solvent may also contain a buffer such as phosphate citrate, sodium phosphate, or potassium phosphate, or, in certain embodiments, other such buffers known to those of skill in the art at approximately neutral pH. Subsequent sterile filtration of the solution followed by lyophilization under standard conditions known to those of skill in the art provides an exemplary formulation. In certain embodiments, the resulting solution is dispensed into vials for lyophilization. The lyophilized powder can be stored under appropriate conditions, for example, at about 4°C to room temperature.

[0348] Reconstitution of this lyophilized powder with water for injection provides a formulation for parenteral administration. For reconstitution, the lyophilized powder is added to sterile water or other suitable carrier.

[0349] In certain embodiments, the conjugates provided herein may be formulated for topical administration or application, for example, in the eye, for example, for topical application to the skin and mucous membranes, in the form of gels, creams, and lotions, as well as for application to the eye, or for intracisternal or intrathecal application. Topical administration is also contemplated for transdermal delivery, and for administration to the eye or mucous membranes, or for inhalation therapy. Nasal solutions of the active compound alone or in combination with other pharmaceutically acceptable excipients can also be administered.

[0350] 5.6. Use and Methods In one aspect, provided herein is a method for removing a polypeptide of interest (target protein) from the surface of a cell using a conjugate described herein. In one aspect, provided herein is a method for removing a polypeptide of interest (target protein) from the extracellular environment using a conjugate described herein. For example, in one embodiment, provided herein is a method for removing a polypeptide of interest (target protein) from the surface of a cell by sequestering the target protein in the lysosomes of the cell using a conjugate described herein. In another embodiment, provided herein is a method for removing a polypeptide of interest (target protein) from the extracellular space (extracellular environment) of a cell by sequestering the target protein in the lysosomes of the cell using a conjugate described herein. In another embodiment, provided herein is a method for removing a polypeptide of interest (target protein) from the surface of a cell by sequestering the target protein in the lysosomes of the cell and degrading the target protein using a conjugate described herein. In another embodiment, provided herein is a method for removing a polypeptide of interest (target protein) from the extracellular space (extracellular environment) of a cell by sequestering the target protein in the lysosomes of the cell using a conjugate described herein.

[0351] Removal of a target protein can refer to the reduction or depletion of the target protein from the cell surface or extracellular space, or the extracellular environment, i.e., the reduction or depletion of the amount of the target protein on the cell surface or in the extracellular environment.

[0352] In one aspect, provided herein is a method for sequestering a polypeptide of interest (target protein) in the lysosomes of a cell using the conjugates described herein. In one aspect, provided herein is a method for sequestering a polypeptide of interest (target protein) in the lysosomes of a cell using the conjugates described herein and degrading the polypeptide of interest.

[0353] In one aspect, provided herein are methods for degrading a polypeptide of interest (target protein) using the conjugates described herein.

[0354] In one aspect, provided herein is a method for depleting a polypeptide of interest (target protein) described herein by degradation via the lysosomal pathway of a cell.

[0355] In another aspect, the present invention provides a method for depleting the polypeptide of interest (target protein) described herein by administering to a subject in need thereof an effective amount of the conjugate or pharmaceutically acceptable salt described herein, or the pharmaceutical composition described herein.In certain embodiments, the subject is a mammal (e.g., human).

[0356] In certain embodiments, the target protein is a VEGF protein, an EGFR protein, a VEGFR protein, a PD-L1 protein, an FGFR2 protein, or an FGFR3 protein.

[0357] In another aspect, provided herein is a method of treating a disease or disorder by administering to a subject, e.g., a human in need thereof, an effective amount of a conjugate or pharmaceutically acceptable salt described herein, or a pharmaceutical composition described herein.

[0358] The terms "administer," "administration," or "administering" refer to the act of injecting or otherwise physically delivering a substance (e.g., a conjugate or pharmaceutical composition provided herein) to a subject or patient (e.g., a human), for example, by mucosal, topical, intradermal, parenteral, intravenous, intramuscular delivery, and / or any other method of physical delivery described herein or known in the art. In certain embodiments, administration is by intravenous infusion.

[0359] The term "effective amount" or "therapeutically effective amount" refers to an amount of a therapeutic agent (e.g., a conjugate or pharmaceutical composition provided herein) sufficient to treat, diagnose, prevent, delay the onset of, reduce the severity and / or duration of, and / or ameliorate a given disease, disorder, or condition, and / or its associated symptoms. These terms also encompass the amount necessary to alleviate, slow, or ameliorate the progression or progression of a given disease, alleviate, slow, or ameliorate the recurrence, development, or onset of a given disease, and / or improve or enhance the prophylactic or therapeutic effect(s) of another therapy, or act as a bridge to another therapy. In some embodiments, "effective amount" as used herein also refers to the amount of a conjugate described herein to achieve a particular result.

[0360] In certain embodiments, when the disorder or disease is cancer, an "effective amount" or "therapeutically effective amount" refers to the amount of a conjugate or pharmaceutical composition provided herein that, when administered to a human suffering from cancer, is sufficient to treat the cancer. "Treatment" or "treatment" of cancer includes one or more of the following: (1) limiting / inhibiting cancer growth, e.g., limiting its occurrence; (2) reducing / preventing the spread of cancer, e.g., reducing / preventing metastasis; (3) Reducing cancer, e.g., causing cancer regression; (4) reducing / preventing cancer recurrence, and (5) Relieving cancer symptoms.

[0361] The terms "subject" and "patient" are used interchangeably. A subject may be a mammal, such as a non-primate (e.g., cows, pigs, horses, cats, dogs, goats, rabbits, rats, mice, etc.) or a primate (e.g., monkeys and humans), e.g., a human. In certain embodiments, the subject is a mammal, e.g., a human, diagnosed with a disease or disorder provided herein. In another embodiment, the subject is a mammal, e.g., a human, at risk of developing a disease or disorder provided herein. In certain embodiments, the subject is a human.

[0362] The term "therapy(s)" can refer to any protocol(s), method(s), composition(s), formulation(s), and / or agent(s) that can be used in the prevention, treatment, management, or amelioration of a disease or disorder or a symptom thereof (e.g., a disease or disorder provided herein, or one or more symptoms or conditions associated therewith). In certain embodiments, the term "therapy(s)" refers to drug therapy, adjuvant therapy, radiation, surgery, biological therapy, supportive therapy, and / or other therapies useful in the treatment, management, prevention, or amelioration of a disease or disorder or one or more symptoms thereof. In certain embodiments, the term "therapy" refers to a therapy other than a conjugate described herein or a pharmaceutical composition thereof.

[0363] In certain embodiments, the disease or disorder is treated by depletion of the target protein by degradation via the lysosomal pathway.

[0364] In certain embodiments, the disease or disorder is treated by depletion of certain proteins, e.g., soluble proteins, e.g., secreted proteins, cell surface proteins (e.g., cell surface receptor proteins, e.g., tyrosine kinase receptors, soluble cytokine receptors, and immune checkpoint receptors, e.g., EGFR, VEGFR, FGFR, and PD-L1), lectins, complement, lipoproteins, transport proteins, MHC class I and class II molecules, cytokines, chemokines, and / or receptors, or fragments or subunits of any of the foregoing.

[0365] In certain embodiments, the disease or disorder is cancer.

[0366] In certain embodiments, the cancer is selected from the group consisting of bladder cancer, breast cancer, cervical cancer, cholangiocarcinoma, endometrial cancer, hepatocellular carcinoma, renal cancer, melanoma, myeloid neoplasms, non-small cell lung cancer (NSCLC), Ewing's sarcoma, and Hodgkin's lymphoma.

[0367] In certain embodiments, the cancer is a solid tumor.

[0368] In certain embodiments, the disease or disorder is an inflammatory or autoimmune disease.

[0369] In certain embodiments, the disease or disorder is an inflammatory disease.

[0370] In certain embodiments, the disease or disorder is an autoimmune disease. In certain embodiments, the disease or disorder is a viral disease. Optionally, the viral disease is hepatitis B.

[0371] 5.7.Definition It is to be understood that the present disclosure is not limited to particular embodiments described, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present disclosure will be limited only by the appended claims.

[0372] Unless otherwise defined, 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. Any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of embodiments of the present disclosure.

[0373] It should be noted that, as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to a "compound" includes a single compound as well as a combination of two or more compounds, reference to a "substituent" includes a single substituent as well as two or more substituents, etc.

[0374] In describing and claiming the present invention, certain terminology will be used in accordance with the definitions set out below. It will be understood that the definitions provided herein are not intended to be mutually exclusive. Accordingly, some chemical moieties may be included within more than one definition of a term.

[0375] As used herein, the phrases "for example," "for instance," "such as," or "including" are intended to introduce examples that further clarify a more general subject matter. These examples are presented solely as an aid in understanding the disclosure and are not intended to be limiting in any way.

[0376] The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein should be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. Further, the publication dates provided may be different from the actual publication dates, which may need to be independently confirmed.

[0377] The terms "protein" and "polypeptide" are used interchangeably. A protein may include moieties other than amino acids (e.g., it may be a glycoprotein, etc.) and / or may be otherwise processed or modified. Those of skill in the art will understand that a "protein" may refer to an entire protein chain (with or without a signal sequence) produced by a cell, or a protein portion thereof. Those of skill in the art will understand that a protein may comprise multiple protein chains, for example, linked noncovalently or covalently, e.g., by one or more disulfide bonds, or associated by other means. In certain embodiments, a polypeptide may exist as a single chain or as two or more associated chains, e.g., as a multimer, e.g., a dimer, a trimer. The term also encompasses amino acid polymers that have been modified, either naturally or by intervention, for example, by disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or other manipulation or modification. Also included within the definition are polypeptides containing one or more analogs of an amino acid, including, but not limited to, unnatural amino acids, as well as other modifications known in the art. Polypeptides may contain L-amino acids, D-amino acids, or both, and may contain any of a variety of amino acid modifications or analogs known in the art. Useful modifications include, for example, terminal acetylation, amidation, methylation, and the like. In some embodiments, proteins may contain natural amino acids, unnatural amino acids, synthetic amino acids, and combinations thereof. In some embodiments, proteins are antibodies, antibody fragments, biologically active portions thereof, and / or characteristic portions thereof.

[0378] The terms "antibody" and "immunoglobulin" are terms of art and may be used interchangeably herein in their broadest sense, including a specific type of immunoglobulin molecule that contains one or more antigen-binding domains that specifically bind to an antigen or epitope.

[0379] In certain embodiments, an isolated antibody (e.g., a monoclonal antibody) described herein, or an antigen-binding fragment thereof that specifically binds to a protein of interest, e.g., EGFR, is conjugated to one or more lysosomal targeting moieties, e.g., via a linker.

[0380] An "antigen" is a moiety or molecule containing an epitope to which an antibody can specifically bind. Thus, an antibody also specifically binds to an antigen. In certain embodiments, the antigen to which the antibodies described herein bind is a protein of interest, such as EGFR (e.g., human EGFR), or a fragment thereof, or, for example, the extracellular domain of EGFR (e.g., human EGFR).

[0381] "Epitope" is a term known in the art and refers to a localized region of an antigen to which an antibody can specifically bind. An epitope may be a linear epitope of contiguous amino acids or may include two or more non-contiguous regions of amino acids of the antigen.

[0382] The terms "binds," "binds to," "specifically binds," or "specifically binds to," in the context of antibody binding, refer to an antibody that binds to an antigen (e.g., an epitope) as such binding is understood by one of skill in the art. For example, a molecule that specifically binds to an antigen may generally bind to other polypeptides with lower affinity, as measured, for example, by immunoassay, Biacore™, KinExA 3000 instrument (Sapidyne Instruments, Boise, ID), or other assays known in the art. In certain embodiments, a molecule that specifically binds to an antigen refers to the affinity (K) with which the molecule binds to another antigen. d ) at least 2, 2.5, 3, or 4 logs lower than dIn another specific embodiment, a molecule that specifically binds to an antigen does not cross-react with other proteins. In another specific embodiment, where EGFR is the protein of interest, a molecule that specifically binds to an antigen does not cross-react with other non-EGFR proteins.

[0383] Antibodies include, in particular, full-length antibodies (e.g., intact immunoglobulins), antibody fragments, monoclonal antibodies, polyclonal antibodies, recombinantly produced antibodies, monospecific antibodies, multispecific antibodies (including bispecific antibodies), human antibodies, humanized antibodies, chimeric antibodies, synthetic antibodies, tetrameric antibodies comprising two heavy chain and two light chain molecules, antibody light chain monomers, antibody heavy chain monomers, antibody light chain dimers, antibody heavy chain dimers, antibody light chain / antibody heavy chain pairs, antibodies having two light chain / heavy chain pairs (e.g., identical pairs), antibody fragments, and the like. Examples of antibodies include, but are not limited to, intrabodies, heteroconjugate antibodies, single domain antibodies, monovalent antibodies, bivalent antibodies (including monospecific or bispecific bivalent antibodies), single chain antibodies or single chain Fvs (scFvs), camelized antibodies, affibodies, Fab fragments, F(ab') fragments, F(ab')2 fragments, disulfide-linked Fvs (sdFvs), anti-idiotypic (anti-Id) antibodies (including, for example, anti-anti-Id antibodies), and epitope-binding fragments of any of the above.

[0384] An antibody may be of any type (e.g., IgG, IgE, IgM, IgD, IgA, or IgY), any class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, or IgA2), or any subclass (e.g., IgG2a or IgG2b) of immunoglobulin molecule. In certain embodiments, the antibodies described herein are IgG antibodies (e.g., human IgG), or classes thereof (e.g., human IgG1, IgG2, IgG3, or IgG4), or subclasses thereof.

[0385] In certain embodiments, the antibody is a four-chain antibody unit comprising two heavy (H) chain / light (L) chain pairs, wherein the amino acid sequences of the H chains are identical and the amino acid sequences of the L chains are identical. In certain embodiments, the H chain and L chain comprise a constant region, e.g., a human constant region. In even more particular embodiments, the L chain constant region of such an antibody is a kappa or lambda light chain constant region, e.g., a human kappa or lambda light chain constant region. In another specific embodiment, the H chain constant region of such an antibody comprises a gamma heavy chain constant region, e.g., a human gamma heavy chain constant region. In certain embodiments, such an antibody comprises an IgG constant region, e.g., a human IgG constant region.

[0386] The term "constant region" or "constant domain" is a well-known antibody term in the art (sometimes referred to as "Fc") and refers to portions of antibodies, e.g., the carboxyl-terminal portions of the light and / or heavy chains, that are not directly involved in binding the antibody to an antigen, but may exhibit various effector functions, such as interaction with Fc receptors. These terms refer to portions of immunoglobulin molecules that generally have more conserved amino acid sequences compared to immunoglobulin variable domains.

[0387] The term "heavy chain" when used in reference to an antibody can refer to any of the different types, e.g., alpha (α), delta (δ), epsilon (ε), gamma (γ), and mu (μ), which give rise to the IgA, IgD, IgE, IgG, and IgM classes of antibodies, including the IgG subclasses, e.g., IgG1, IgG2, IgG3, and IgG4, respectively, based on the amino acid sequence of the constant domain.

[0388] The term "light chain" when used in reference to an antibody can refer to any of different types, e.g., lambda (λ) or kappa (κ), based on the amino acid sequence of its constant domain. Light chain amino acid sequences are well known in the art. In certain embodiments, the light chain is a human light chain.

[0389] The term "monoclonal antibody" is a well-known term of art that refers to an antibody obtained from a homogeneous or substantially homogeneous population of antibodies. The term "monoclonal" is not limited to any particular method for producing the antibody. Generally, a population of monoclonal antibodies can be produced by a cell, a population of cells, or a cell line. In certain embodiments, a "monoclonal antibody" as used herein is an antibody produced by a single cell (e.g., a hybridoma or a host cell producing a recombinant antibody), which specifically binds to an epitope, as measured, for example, by ELISA or other antigen-binding or competitive binding assays known in the art or in the Examples provided herein. In certain embodiments, a monoclonal antibody may be a chimeric or humanized antibody. In certain embodiments, a monoclonal antibody is a monovalent antibody or a multivalent (e.g., bivalent) antibody. In certain embodiments, a monoclonal antibody is a monospecific or multispecific antibody (e.g., a bispecific antibody).

[0390] The term "variable region" or "variable domain" refers to a portion of an antibody, generally a portion of either the light or heavy chain, usually from the amino terminus, approximately 110-120 amino acids in the mature heavy chain and approximately 90-100 amino acids in the mature light chain. The variable region comprises complementarity-determining regions (CDRs) flanked by framework regions (FRs). Generally, the spatial orientation of the CDRs and FRs, from N-terminus to C-terminus, is as follows: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4. Without wishing to be bound by any particular mechanism or theory, it is believed that the CDRs of the light and heavy chains are primarily responsible for the interaction of the antibody with the antigen and the specificity of the antibody for the epitope. In certain embodiments, the numbering of the amino acid positions of the antibodies described herein is according to the EU index, such as in Kabat et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, USDapartment of Health and Human Services, NIH Publication No. 91-3242. In certain embodiments, the variable regions are human variable regions.

[0391] In certain embodiments, the CDRs of the antibody are numbered according to (i) the Kabat numbering system (Kabat et al. (1971) Ann. NY Acad. Sci. 190:382-391 and Kabat et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242), or (ii) the Chothia numbering scheme (e.g., Chothia and Lesk, 1987, J. Mol. Biol., 196:901-917; Al-Lazikani et al., 1997, J. Mol. Biol., 273:927-948; Chothia et al., 1992, J. Mol. Biol., 227:799-817; Tramontano et al., 1992, J. Mol. Biol., 227:799-817), herein referred to as "Chothia CDRs." al., 1990, J. Mol. Biol. 215(1):175-82; U.S. Patent No. 7,709,226; and Martin, A., “Protein Sequence and Structure Analysis of Antibody Variable Domains,” in Antibody Engineering, Kontermann and Dubel, eds., Chapter 31, pp. 422-439, Springer-Verlag, Berlin (2001)); or (iii) the ImMunoGeneTics (IMGT) numbering system, as described, for example, in Lefranc, 1999, The Immunologist, 7:132-136 and Lefranc et al., 1999, Nucleic Acids Res., 27:209-212 (“IMGT CDRs”); or (iv) the ImMunoGeneTics (IMGT) numbering system, as described, for example, in MacCallum et al., 1996, J. Mol. Biol., 262: The AbM CDRs can be determined according to the AbM numbering system, set forth in pp. 732-745, herein referred to as "AbM CDRs."See also, for example, Martin, A., "Protein Sequence and Structure Analysis of Antibody Variable Domains," in Antibody Engineering, Kontermann and Dubel, eds., Chapter 31, pp. 422-439, Springer-Verlag, Berlin (2001); or (v) the contact numbering system, referred to herein as "contact CDRs," where the contact definitions are based on analysis of available complex crystal structures (bioinf.org.uk / abs) (see, for example, MacCallum et al., 1996, J. Mol. Biol., 262:732-745).

[0392] The terms "full length antibody," "intact antibody," and "whole antibody" are used interchangeably herein to refer to an antibody in a substantially intact form, and not an antibody fragment as defined below. These terms particularly refer to antibodies having heavy chains that include an Fc region.

[0393] An "antibody fragment" comprises only a portion of an intact antibody, which portion retains at least one, two, three, or most or all of the functions normally associated with that portion when present in an intact antibody. In one embodiment, an antibody fragment comprises the antigen-binding site of an intact antibody and thus retains the ability to bind to an antigen. In another embodiment, an antibody fragment, such as an antibody fragment comprising an Fc region, retains at least one of the biological functions normally associated with the Fc region when present in an intact antibody. Such functions may include FcRn binding, antibody half-life modulation, conjugation function, and complement fixation. In another embodiment, an antibody fragment is a monovalent antibody with an in vivo half-life substantially similar to that of an intact antibody. For example, such an antibody fragment may comprise a single antigen-binding arm linked to an Fc sequence, which may confer in vivo stability to the fragment. Antibody fragments suitable for use in the compounds of the present disclosure include, for example, Fv fragments, Fab fragments, F(ab')2 fragments, Fab' fragments, scFv (sFv) fragments, and scFv-Fc fragments.

[0394] "Polynucleotide" or "nucleic acid" are used interchangeably herein and refer to a polymeric form of nucleotides of any length, either deoxyribonucleotides or ribonucleotides, or their analogs. Polynucleotides can have any three-dimensional structure and can perform any function, known or unknown. Non-limiting examples of polynucleotides include genes, gene fragments, exons, introns, messenger RNA (mRNA), transfer RNA, ribosomal RNA, ribozymes, cDNA, recombinant polynucleotides, branched polynucleotides, plasmids, vectors, isolated DNA of any sequence, control regions, isolated RNA of any sequence, nucleic acid probes, and primers. Nucleic acid molecules can be linear or circular. The nucleotides can be deoxyribonucleotides, ribonucleotides, modified nucleotides or bases, and / or their analogs, or any substrate that can be incorporated into a polymer by DNA or RNA polymerase or by a synthetic reaction. Polynucleotides can also contain modified nucleotides, such as methylated nucleotides and their analogs. The nucleic acid molecule can be an aptamer.

[0395] The term "purified" refers to the isolation of a substance (compound, polynucleotide, protein, polypeptide, polypeptide composition) of interest such that the substance constitutes a majority of a sample containing said substance. Typically, a substantially purified component of a sample constitutes 50%, 80%-85%, 90-99%, e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the sample. Techniques for purifying polynucleotides, polypeptides, and viral particles of interest are well known in the art and include, for example, ion exchange chromatography, affinity chromatography, and sedimentation by density.

[0396] The terms "treatment," "treating," and the like refer to obtaining a desired pharmacological and / or physiological effect, such as reducing tumor burden. The effect may be prophylactic, in that a disease or its symptoms are completely or partially prevented, and / or therapeutic, in that the disease and / or side effects caused by the disease are partially or completely cured. As used herein, "treatment" encompasses any treatment of a disease in a mammal, particularly a human, and includes (a) preventing the onset of the disease or symptoms of the disease in a subject who is susceptible to the disease but has not yet been diagnosed with the disease (including, for example, diseases that may be associated with or caused by the primary disease, such as liver fibrosis that may occur in the context of chronic HCV infection); (b) inhibiting the disease, i.e., halting the progression of the disease; and (c) relieving the disease, i.e., causing regression of the disease (e.g., reducing tumor burden).

[0397] The terms "individual," "host," "subject," and "patient" are used interchangeably herein and refer to animals including, but not limited to, humans and non-human primates, including apes and humans; rodents, including rats and mice, cows, horses, sheep, cats, dogs, etc. "Mammal" means a member or members of any mammalian species, including, for example, dogs, cats, horses, cows, sheep, rodents, etc., and primates, e.g., non-human primates, and humans. Non-human animal models, e.g., mammals, e.g., non-human primates, rats, rabbits, etc., may be used in experimental studies.

[0398] "Therapeutically effective amount" or "effective amount" means the amount of a compound that, when administered to a mammal or other subject for treating a disease, illness, or disorder, is sufficient to effect such treatment for the disease, illness, or disorder. The "therapeutically effective amount" will vary depending on the compound, the disease and its severity, and the age, weight, etc., of the subject being treated.

[0399] Unless otherwise specified, in cases where a compound can exist in alternative tautomeric, regioisomeric, and / or stereoisomeric forms, all alternative isomers are intended to be encompassed within the scope of the claimed subject matter. For example, if a compound is described as a particular optical isomer, D- or L-, both optical isomers are intended to be encompassed herein. For example, if a compound is described as having one of two tautomeric forms, both tautomers are intended to be encompassed herein. Thus, the compounds provided herein may be enantiomerically pure or may be mixtures of stereoisomers or diastereomers. The compounds provided herein may contain chiral centers. Such chiral centers may be in either the (R) or (S) configuration, or may be mixtures thereof. The chiral centers of the compounds provided herein may undergo epimerization in vivo. Thus, one of skill in the art will recognize that, for compounds that undergo epimerization in vivo, administering a compound in its (R) form is equivalent to administering the compound in its (S) form.

[0400] The present disclosure also encompasses all suitable isotopic variants of the compounds of the present disclosure, whether radioactive or not.Isotopic variants of the compounds of the present disclosure are understood to mean compounds in which at least one atom in the compounds of the present disclosure is replaced with another atom having the same atomic number but an atomic mass different from the atomic mass that is usually or predominantly present in nature.Examples of isotopes that can be incorporated into compounds of the present disclosure include hydrogen, carbon, nitrogen, oxygen, fluorine, chlorine, bromine, and iodine, for example: 2 H (deuterium), 3 H (tritium), 13 C. 14 C. 15 N, 17 O. 18 O. 18 F, 36 Cl, 82 Br, 123 I, 124 I, 125 I, 129 I, and131 I is an isotope of I. Certain isotopic variants of the compounds of the present disclosure, particularly those incorporating one or more radioisotopes, may be useful, for example, in studying the mechanism of action or distribution of the active compound in the body. 3 H, 14 C, and / or 18 Compounds labeled with F isotopes are suitable for this purpose. Additionally, the incorporation of isotopes, such as deuterium, may lead to certain therapeutic advantages as a result of greater metabolic stability of the compound, such as an increased half-life in the body or a reduced required active dose. In some embodiments, hydrogen atoms in the compounds described herein can be replaced with deuterium atoms. In certain embodiments, "deuterated" as applied to a chemical group refers to a chemical group that is isotopically enriched with deuterium in an amount substantially greater than its natural abundance, unless otherwise indicated. Isotopic variants of the compounds according to the present disclosure can be prepared in various ways, for example, by using corresponding isotopic modifications of certain reactants and / or starting compounds therein in the methods and examples described below.

[0401] Therefore, any of the embodiments described herein are intended to encompass salts, single stereoisomers, mixtures of stereoisomers, and / or isotopic forms of the compounds.

[0402] "Pharmaceutically acceptable excipient," "pharmaceutically acceptable diluent," "pharmaceutically acceptable carrier," and "pharmaceutically acceptable adjuvant" refer to excipients, diluents, carriers, and adjuvants that are generally safe, non-toxic, and not biologically or otherwise undesirable and are useful in the preparation of pharmaceutical compositions, including excipients, diluents, carriers, and adjuvants that are acceptable for veterinary as well as human pharmaceutical use. As used in the specification and claims, "pharmaceutically acceptable excipients, diluents, carriers, and adjuvants" includes both one and more of such excipients, diluents, carriers, and adjuvants.

[0403] "Pharmaceutical composition" is meant to encompass compositions suitable for administration to a subject, such as a mammal, particularly a human. Generally, a "pharmaceutical composition" is sterile and preferably free of contaminants that could induce an undesirable reaction in the subject (e.g., the compound(s) in the pharmaceutical composition are of pharmaceutical grade). Pharmaceutical compositions can be designed for administration to a subject or patient in need of such composition via many different routes of administration, including oral, buccal, rectal, parenteral, intraperitoneal, intradermal, intratracheal, intramuscular, subcutaneous, etc.

[0404] The term "pharmaceutically acceptable" means approved by a federal or state regulatory agency or listed in the United States Pharmacopoeia, the European Pharmacopoeia, or other generally recognized pharmacopeia for use in animals, and more particularly in humans.

[0405] The term "pharmaceutically acceptable salt" refers to a salt suitable for use in contact with the tissues of humans and lower animals without causing undue toxicity, irritation, allergic reactions, etc. Pharmaceutically acceptable salts are well known in the art. For example, S.M. Berge, et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66:1-19 (1977). Such salts can be prepared in situ during the final isolation and purification of the conjugate compound, or separately by reacting the free base functional group of the compound with an appropriate organic acid. Examples of pharmaceutically acceptable salts include, but are not limited to, non-toxic acid addition salts or salts of amino groups formed with inorganic acids.

[0406] "Acyl" refers to the groups HC(O)-, alkyl-C(O)-, substituted alkyl-C(O)-, alkenyl-C(O)-, substituted alkenyl-C(O)-, alkynyl-C(O)-, substituted alkynyl-C(O)-, cycloalkyl-C(O))-, substituted cycloalkyl-C(O)-, cycloalkenyl-C(O)-, substituted cycloalkenyl-C(O)-, aryl-C(O)-, substituted aryl-C(O)-, heteroaryl-C(O)-, "C(O)-," "heterocyclyl-C(O)-," and "substituted heterocyclyl-C(O)-," where alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocycle, and substituted heterocycle are as defined herein. For example, acyl includes the "acetyl" group CHC(O)-.

[0407] The term "alkyl" refers to a branched or unbranched saturated hydrocarbon group (i.e., a monoradical) typically (but not necessarily) containing from 1 to about 24 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, octyl, decyl, and the like, as well as cycloalkyl groups such as cyclopentyl and cyclohexyl. Alkyl groups herein generally (but not necessarily) may contain from 1 to about 18 carbon atoms, and such groups may contain from 1 to about 12 carbon atoms. The term "lower alkyl" refers to an alkyl group of 1 to 6 carbon atoms. "Substituted alkyl" refers to an alkyl substituted with one or more substituents, including instances in which two hydrogen atoms are replaced from the same carbon atom in an alkyl substituent, such as in a carbonyl group (i.e., a substituted alkyl group may contain a -C(=O)- moiety). The terms "heteroatom-containing alkyl" and "heteroalkyl" refer to an alkyl substituent in which at least one carbon atom is replaced with a heteroatom, as described in more detail below. Unless otherwise indicated, the terms "alkyl" and "lower alkyl" include straight-chain, branched-chain, cyclic, unsubstituted, substituted, and / or heteroatom-containing alkyl or lower alkyl, respectively.

[0408] The term "substituted alkyl" refers to an alkyl group, as defined herein, wherein one or more carbon atoms in the alkyl chain are optionally replaced with a heteroatom, such as -O-, -N-, -S-, -S(O)n- (n is 0-2), -NR- (wherein R is hydrogen or alkyl), and includes alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, oxo, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thio is meant to include the above alkyl groups having 1 to 5 substituents selected from the group consisting of all, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO-aryl, -SO-heteroaryl, -SO2-alkyl, -SO2-aryl, -SO2-heteroaryl, and NRaRb (wherein R' and R'' are the same or different and are selected from hydrogen, optionally substituted alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl, and heterocycle).

[0409] The term "alkenyl" refers to a straight-chain, branched-chain, or cyclic hydrocarbon group containing at least one double bond and having 2 to about 24 carbon atoms, such as ethenyl, n-propenyl, isopropenyl, n-butenyl, isobutenyl, octenyl, decenyl, tetradecenyl, hexadecenyl, eicosenyl, tetracosenyl, and the like. Alkenyl groups herein generally (although not necessarily) may contain from 2 to about 18 carbon atoms, e.g., from 2 to 12 carbon atoms. The term "lower alkenyl" refers to an alkenyl group of 2 to 6 carbon atoms. The term "substituted alkenyl" refers to an alkenyl substituted with one or more substituents, and the terms "heteroatom-containing alkenyl" and "heteroalkenyl" refer to an alkenyl in which at least one carbon atom is replaced with a heteroatom. Unless otherwise indicated, the terms "alkenyl" and "lower alkenyl" include straight-chain, branched, cyclic, unsubstituted, substituted, and / or heteroatom-containing alkenyl and lower alkenyl, respectively.

[0410] The term "alkynyl" refers to a straight- or branched-chain hydrocarbon group containing 2 to 24 carbon atoms, such as ethynyl, n-propynyl, and the like, and containing at least one triple bond. Alkynyl groups herein generally (although not necessarily) may contain from 2 to about 18 carbon atoms, although such groups may also contain from 2 to 12 carbon atoms. The term "lower alkynyl" contemplates an alkynyl group of 2 to 6 carbon atoms. The term "substituted alkynyl" refers to an alkynyl substituted with one or more substituents, and the terms "heteroatom-containing alkynyl" and "heteroalkynyl" refer to an alkynyl having at least one carbon atom replaced with a heteroatom. Unless otherwise indicated, the terms "alkynyl" and "lower alkynyl" include straight-chain, branched, unsubstituted, substituted, and / or heteroatom-containing alkynyl and lower alkynyl, respectively.

[0411] The term "alkoxy" refers to an alkyl group attached through a single, terminal ether linkage. That is, an "alkoxy" group can be represented as -O-alkyl, where alkyl is as defined above. A "lower alkoxy" group refers to an alkoxy group having from 1 to 6 carbon atoms, and includes, for example, methoxy, ethoxy, n-propoxy, isopropoxy, t-butyloxy, and the like. Substituents herein identified as "C1-C6 alkoxy" or "lower alkoxy" can contain, for example, 1 to 3 carbon atoms; as a further example, such substituents can contain 1 or 2 carbon atoms (i.e., methoxy and ethoxy).

[0412] The term "substituted alkoxy" refers to the groups substituted alkyl-O-, substituted alkenyl-O-, substituted cycloalkyl-O-, substituted cycloalkenyl-O-, and substituted alkynyl-O-, where substituted alkyl, substituted alkenyl, substituted cycloalkyl, substituted cycloalkenyl, and substituted alkynyl are as defined herein.

[0413] The term "aryl," unless otherwise specified, generally (though not necessarily) refers to an aromatic substituent containing 5 to 30 carbon atoms and containing a single aromatic ring or multiple aromatic rings fused, directly linked, or indirectly linked together (such that different aromatic rings are bonded to a common group, such as a methylene or ethylene moiety). An aryl group may contain, for example, 5 to 20 carbon atoms; as a further example, an aryl group may contain 5 to 12 carbon atoms. For example, an aryl group may contain one aromatic ring or two or more fused or linked aromatic rings (i.e., biaryl, aryl-substituted aryl, etc.). Examples include phenyl, naphthyl, biphenyl, diphenyl ether, diphenylamine, benzophenone, etc. "Substituted aryl" refers to an aryl moiety substituted with one or more substituents, and the terms "heteroatom-containing aryl" and "heteroaryl" refer to an aryl substituent in which at least one carbon atom has been replaced with a heteroatom, as described in more detail below. Aryl includes stable cyclic, heterocyclic, polycyclic, and polyheterocyclic unsaturated C-C 14 Moieties are intended to include, but are not limited to, phenyl, biphenyl, naphthyl, pyridyl, furyl, thiophenyl, imidazolyl, pyrimidinyl, and oxazolyl, which may be further substituted with 1 to 5 members selected from the group consisting of hydroxy, C1-C8 alkoxy, C1-C8 branched or straight chain alkyl, acyloxy, carbamoyl, amino, N-acylamino, nitro, halogen, trifluoromethyl, cyano, and carboxyl (see, e.g., Katritzky, Handbook of Heterocyclic Chemistry). Unless otherwise indicated, the term "aryl" includes unsubstituted, substituted, and / or heteroatom-containing aromatic substituents.

[0414] The term "aralkyl" refers to an alkyl group having an aryl substituent, and the term "alkaryl" refers to an aryl group having an alkyl substituent, where "alkyl" and "aryl" are defined above. Generally, aralkyl and alkaryl groups herein contain 6 to 30 carbon atoms. Aralkyl and alkaryl groups can, for example, contain 6 to 20 carbon atoms, and as a further example, such groups can contain 6 to 12 carbon atoms.

[0415] The term "alkylene" refers to a diradical alkyl group. Unless otherwise indicated, such groups include saturated hydrocarbon chains containing 1 to 24 carbon atoms, which may be substituted or unsubstituted, may contain one or more alicyclic groups, and may contain heteroatoms. "Lower alkylene" refers to an alkylene linkage containing 1 to 6 carbon atoms. Examples include methylene (--CH--), ethylene (--CHCH--), propylene (--CHCHCH--), 2-methylpropylene (--CH--CH(CH)--CH--), hexylene (--(CH)--), and the like.

[0416] Similarly, the terms "alkenylene," "alkynylene," "arylene," "aralkylene," and "alkarylene" refer to the diradical alkenyl, alkynyl, aryl, aralkyl, and alkaryl groups, respectively.

[0417] The term "amino" refers to the group -NRR', where R and R' are independently hydrogen or non-hydrogen substituents, including, for example, alkyl, aryl, alkenyl, aralkyl, and substituted and / or heteroatom-containing variants thereof.

[0418] The terms "halo" and "halogen" are used in the conventional sense to refer to a chloro, bromo, fluoro, or iodo substituent.

[0419] "Carboxyl", "carboxy", or "carboxylate" refers to -CO2H or its salts.

[0420] "Cycloalkyl" refers to cyclic alkyl groups of 3 to 10 carbon atoms having single or multiple cyclic rings, including fused, bridged, and spiro ring systems. Examples of suitable cycloalkyl groups include, for example, adamantyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, and the like. Such cycloalkyl groups include, for example, single ring structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, and the like, or multiple ring structures such as adamantanyl.

[0421] The term "substituted cycloalkyl" refers to a cycloalkyl group having from 1 to 5 substituents, or from 1 to 3 substituents, selected from alkyl, substituted alkyl, alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, oxo, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -SO2-alkyl, -SO2-substituted alkyl, -SO2-aryl, and -SO2-heteroaryl.

[0422] The term "heteroatom-containing," as in "heteroatom-containing alkyl group" (also referred to as "heteroalkyl" group) or "heteroatom-containing aryl group" (also referred to as "heteroaryl" group), refers to a molecule, bond, or substituent in which one or more carbon atoms have been replaced with an atom other than carbon, such as nitrogen, oxygen, sulfur, phosphorus, or silicon, typically nitrogen, oxygen, or sulfur. Similarly, the term "heteroalkyl" refers to an alkyl substituent containing a heteroatom; the term "heterocycloalkyl" refers to a cycloalkyl substituent containing a heteroatom; the terms "heterocyclic" or "heterocycle" refer to cyclic substituents containing heteroatoms; the terms "heteroaryl" and "heteroaromatic" refer to "aryl" and "aromatic" substituents, respectively, that contain heteroatoms; and so forth. Examples of heteroalkyl groups include alkoxyaryl, alkylsulfanyl-substituted alkyls, N-alkylated aminoalkyls, and the like. Examples of heteroaryl substituents include pyrrolyl, pyrrolidinyl, pyridinyl, quinolinyl, indolyl, furyl, pyrimidinyl, imidazolyl, 1,2,4-triazolyl, tetrazolyl, and the like; examples of heteroatom-containing alicyclic groups are pyrrolidino, morpholino, piperazino, piperidino, tetrahydrofuranyl, and the like.

[0423] "Heteroaryl" refers to an aromatic group having 1 to 15 carbon atoms in the ring, e.g., 1 to 10 carbon atoms, and 1 to 10 heteroatoms selected from the group consisting of oxygen, nitrogen, and sulfur. Such heteroaryl groups can have a single ring (such as pyridinyl, imidazolyl, or furyl) or multiple condensed rings in the ring system (e.g., in groups such as indolizinyl, quinolinyl, benzofuran, benzimidazolyl, or benzothienyl), provided that at least one ring in the ring system is aromatic, provided that the point of attachment is through an atom in the aromatic ring. In certain embodiments, the nitrogen and / or sulfur ring atom(s) of a heteroaryl group are optionally oxidized to provide N-oxide (N→O), sulfinyl, or sulfonyl moieties. The term includes, by way of example, pyridinyl, pyrrolyl, indolyl, thiophenyl, and furanyl. Unless otherwise constrained in the definition of a heteroaryl substituent, such heteroaryl groups are optionally substituted with 1 to 5 substituents, or 1 to 3 substituents, selected from acyloxy, hydroxy, thiol, acyl, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl, substituted alkoxy, substituted alkenyl, substituted alkynyl, substituted cycloalkyl, substituted cycloalkenyl, amino, substituted amino, aminoacyl, acylamino, alkaryl, aryl, aryloxy, azido, carboxyl, carboxylalkyl, cyano, halogen, nitro, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy, aminoacyloxy, oxyacylamino, thioalkoxy, substituted thioalkoxy, thioaryloxy, thioheteroaryloxy, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -SO2-alkyl, -SO2-substituted alkyl, -SO2-aryl, and -SO2-heteroaryl, and trihalomethyl.

[0424] The terms "heterocycle," "heterocyclic," and "heterocyclyl" refer to saturated or unsaturated groups having a single ring or multiple fused rings (including fused bridged and spiro ring systems) and having 3 to 15 ring atoms, including 1 to 4 heteroatoms. These ring heteroatoms are selected from nitrogen, sulfur, and oxygen; in fused ring systems, one or more of the rings may be cycloalkyl, heterocycloalkyl, aryl, or heteroaryl, provided that the point of attachment is through a non-aromatic ring. In certain embodiments, the nitrogen and / or sulfur atom(s) of a heterocyclic group are optionally oxidized to provide an N-oxide, -S(O)-, or -SO2- moiety.

[0425] Examples of heterocycles and heteroaryls include azetidine, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, dihydroindole, indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine, naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, phenanthroline, isothiazole, phenazine, isoxazole, phenanthroline, phenanthi ... These include, but are not limited to, phenoxazine, phenothiazine, imidazolidine, imidazoline, piperidine, piperazine, indoline, phthalimide, 1,2,3,4-tetrahydroisoquinoline, 4,5,6,7-tetrahydrobenzo[b]thiophene, thiazole, thiazolidine, thiophene, benzo[b]thiophene, morpholinyl, thiomorpholinyl (also called thiamorpholinyl), 1,1-dioxothiomorpholinyl, piperidinyl, pyrrolidine, tetrahydrofuranyl, and the like.

[0426] Unless otherwise constrained in the definition of a heterocyclic substituent, such heterocyclic groups are optionally substituted with 1 to 5 or 1 to 3 substituents selected from alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, oxo, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -SO2-alkyl, -SO2-substituted alkyl, -SO2-aryl, -SO2-heteroaryl, and fused heterocycle.

[0427] "Hydrocarbyl" refers to a monovalent hydrocarbyl radical, such as an alkyl group, an alkenyl group, or an aryl group, containing 1 to about 30 carbon atoms, including 1 to about 24 carbon atoms, including 1 to about 18 carbon atoms, and including about 1 to 12 carbon atoms, and including straight-chain, branched-chain, cyclic, saturated, and unsaturated species. The hydrocarbyl may be substituted with one or more substituents. The term "heteroatom-containing hydrocarbyl" refers to a hydrocarbyl in which at least one carbon atom has been replaced with a heteroatom. Unless otherwise indicated, the term "hydrocarbyl" should be interpreted as including substituted and / or heteroatom-containing hydrocarbyl moieties.

[0428] "Substituted," as in "substituted hydrocarbyl," "substituted alkyl," "substituted aryl," etc., as mentioned in some of the definitions above, means that at least one hydrogen atom bonded to a carbon (or other) atom in a hydrocarbyl, alkyl, aryl, or other moiety has been replaced with one or more non-hydrogen substituents. Examples of such substituents include, but are not limited to, functional groups and hydrocarbyl moieties, C1-C24 alkyl (including C1-C18 alkyl, further including C1-C12 alkyl, and further including C1-C6 alkyl), C2-C24 alkenyl (including C2-C18 alkyl, further including C2-C12 alkenyl, and further including C2-C6 alkenyl), C2-C24 alkynyl (including C2-C18 alkynyl, further including C2-C12 alkynyl, and further including C2-C6 alkynyl), C5-C30 aryl (including C5-C20 aryl, further including C5-C12 aryl), and C6-C30 aralkyl (including C6-C20 aralkyl, and further including C6-C12 aralkyl). The aforementioned hydrocarbyl moieties may be further substituted with one or more functional groups or additional hydrocarbyl moieties such as those specifically listed. Unless otherwise indicated, any group described herein should be interpreted as including substituted and / or heteroatom-containing moieties in addition to unsubstituted moieties.

[0429] "Sulfonyl" refers to the groups SO-alkyl, SO-substituted alkyl, SO-alkenyl, SO-substituted alkenyl, SO-cycloalkyl, SO-substituted cycloalkyl, SO-cycloalkenyl, SO-substituted cycloalkenyl, SO-aryl, SO-substituted aryl, SO-heteroaryl, SO-substituted heteroaryl, SO-heterocyclic, and SO-substituted heterocyclic, where alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocycle, and substituted heterocycle are as defined herein. Sulfonyl, by way of example, includes methyl-SO-, phenyl-SO-, and 4-methylphenyl-SO-.

[0430] The term "functional group" refers to halo, hydroxyl, sulfhydryl, C1-C24 alkoxy, C2-C24 alkenyloxy, C2-C24 alkynyloxy, C5-C20 aryloxy, acyl (e.g., C2-C24 alkylcarbonyl (-CO-alkyl) and C6-C20 arylcarbonyl (-CO-aryl)), acyloxy (-O-acyl), C2-C24 alkoxycarbonyl (-(CO)-O-alkyl), C6-C20 aryloxycarbonyl (-(CO)-O-aryl), halocarbonyl (-CO)-X (where X is halo). )), C2-C24 alkylcarbonato (-O-(CO)-O-alkyl), C6-C20 arylcarbonato (-O-(CO)-O-aryl), carboxy (-COOH), carboxylato (-COO-), carbamoyl (-(CO)-NH2), monosubstituted C1-C24 alkylcarbamoyl (-(CO)-NH(C1-C24 alkyl)), disubstituted alkylcarbamoyl (-(CO)-N(C1-C24 alkyl)2), monosubstituted arylcarbamoyl (-(CO)-NH-aryl), thiocarbamoyl (-(CS)-NH2), carbami amide (-NH-(CO)-NH2), cyano (-C≡N), isocyano (-N≡C-), cyanato (-OC≡N), isocyanato (-O-N≡C-), isothiocyanato (-SC≡N), azido (-N=N≡N-), formyl (-(CO)-H), thioformyl (-(CS)-H), amino (-NH2), mono- and di-(C1-C24 alkyl) substituted amino, mono- and di-(C5-C20 aryl) substituted amino, C2-C24 alkylamido (-NH-(CO)-alkyl), C5-C20 arylamido (-NH-(CO)-aryl) aryl), imino (-CR=NH (where R=hydrogen, C1-C24 alkyl, C5-C20 aryl, C6-C20 alkaryl, C6-C20 aralkyl, etc.)), alkylimino (-CR=N(alkyl) (where R=hydrogen, alkyl, aryl, alkaryl, etc.)), arylimino (-CR=N(aryl) (where R=hydrogen, alkyl, aryl, alkaryl, etc.)), nitro (-NO2), nitroso (-NO), sulfo (-SO2-OH), sulfonato (-SO2-O-), C1-C24 alkylsulfanyl (-S-alkyl;"Ci-C alkylsulfinyl" refers to chemical groups such as C1-C24 alkylsulfinyl (-(SO)-alkyl), C5-C20 arylsulfinyl (-(SO)-aryl), C1-C24 alkylsulfonyl (-SO2-alkyl), C5-C20 arylsulfonyl (-SO2-aryl), phosphono (-P(O)(OH)2), phosphonato (-P(O)(O-)2), phosphinato (-P(O)(O-)), phospho (-PO2), and phosphino (-PH2), mono- and di-(C1-C24 alkyl)-substituted phosphino, mono- and di-(C5-C20 aryl)-substituted phosphines. Furthermore, the aforementioned functional groups may be further substituted with one or more additional functional groups, or one or more hydrocarbyl moieties, such as those specifically listed above, if the particular group allows.

[0431] "Linkage" or "linker," as in "linking group," "linker moiety," etc., refers to a linking moiety that connects two groups via a covalent bond. The linker may be linear, branched, cyclic, or a single atom. Examples of such linking groups include alkyl, alkenylene, alkynylene, arylene, alkarylene, aralkylene, and linking moieties that contain functional groups, including, but not limited to, amide (-NH-CO-), ureylene (-NH-CO-NH-), imide (-CO-NH-CO-), epoxy (-O-), epithio (-S-), epidioxy (-OO-), carbonyldioxy (-O-CO-O-), alkyldioxy (-O-(CH2)nO-), epoxyimino (-O-NH-), epimino (-NH-), carbonyl (-CO-), and the like. In certain cases, one, two, three, four, or five or more carbon atoms of the linker backbone may be optionally substituted with sulfur, nitrogen, or oxygen heteroatoms. The bonds between the backbone atoms may be saturated or unsaturated, and typically there will be no more than one, two, or three unsaturated bonds in the linker backbone. The linker may include one or more substituents, such as alkyl, aryl, or alkenyl groups. Linkers may include, but are not limited to, poly(ethylene glycol) unit(s) (e.g., —(CH—CH—O)—), ether, thioether, amine, alkyl (e.g., (C1-C 12 ) alkyl) (which may be straight or branched, e.g., methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl), etc.). The linker backbone may comprise a cyclic group, e.g., an aryl, heterocycle, or cycloalkyl group, where two or more atoms, e.g., 2, 3, or 4 atoms, of the cyclic group are included in the backbone. Linkers may be cleavable or non-cleavable. Any convenient arrangement and / or attachment of the linker to the groups to be linked may be used.

[0432] When the term "substituted" appears before a list of possible substituents, it is intended that the term apply to all groups that make up that group. For example, the phrase "substituted alkyl and aryl" should be interpreted as "substituted alkyl and substituted aryl."

[0433] Further to the disclosure herein, the term "substituted," when used to modify a specified group or radical, can also mean that one or more hydrogen atoms of the specified group or radical are each independently replaced with the same or different substituents, as defined below.

[0434] In addition to the groups disclosed for each individual term herein, substituents replacing one or more hydrogens on a saturated carbon atom in a specified group or radical (any two hydrogens on a single carbon can be replaced by ═O, ═NR, etc.) are also included. 70 , =N-OR 70 , =N2, or =S) is, unless otherwise specified, -R 60 , halo, =O, -OR 70 , -SR 70 , -NR 80 R 80 , trihalomethyl, -CN, -OCN, -SCN, -NO, -NO2, =N2, -N3, -SO2R 70 , -SO2O - M + , -SO2OR 70 , -OSO2R 70 , -OSO2O - M + , -OSO2OR 70 , -P(O)(O - )2(M + )2, -P(O)(OR 70 )O - M + , -P(O)(OR 70 )2, -C(O)R 70 , -C(S)R 70 , -C(NR 70 )R 70 , -C(O)O - M + , -C(O)OR 70, -C(S)OR 70 , -C(O)NR 80 R 80 , -C(NR 70 )NR 80 R 80 , -OC(O)R 70 , -OC(S)R 70 , -OC(O)O - M + , -OC(O)OR 70 , -OC(S)OR 70 , -NR 70 C(O)R 70 , -NR 70 C(S)R 70 , -NR 70 CO2 - M + , -NR 70 CO2R 70 , -NR 70 C(S)OR 70 , -NR 70 C(O)NR 80 R 80 , -NR 70 C(NR 70 )R 70 , and -NR 70 C(NR 70 )NR 80 R 80 where R 60 is selected from the group consisting of optionally substituted alkyl, cycloalkyl, heteroalkyl, heterocycloalkylalkyl, cycloalkylalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl; each R 70 are independently hydrogen or R 60 and each R 80 are independently R 70 Or, two R 80 together with the nitrogen atom to which they are attached form a 5-, 6-, or 7-membered heterocycloalkyl, which optionally contains 1 to 4 identical or different additional heteroatoms selected from the group consisting of O, N, and S, wherein N may have —H or C1-C3 alkyl substitution; each M + is a counterion with a net single positive charge.+ are independent, e.g., K + , Na + , Li + Alkaline ions such as; + N(R 60 ) 4; or [Ca 2+ ] 0.5 , [Mg 2+ ] 0.5 , or [Ba 2+ ] 0.5 (The "subscript 0.5" contemplates that one of the counterions to such divalent alkaline earth ions may be an ionized form of a compound of the invention and the other may be a common counterion such as chloride, or a doubly ionized compound disclosed herein may serve as a counterion to such divalent alkaline earth ion, or a doubly ionized compound of the invention may serve as a counterion to such divalent alkaline earth ion.) Specific examples include -NR 80 R 80 is intended to include -NH2, -NH-alkyl, N-pyrrolidinyl, N-piperazinyl, N-methylpiperazin-1-yl, and N-morpholinyl.

[0435] Further to the disclosure herein, the substitution of hydrogen on an unsaturated carbon atom in a "substituted" alkene, alkyne, aryl, and heteroaryl group is represented by -R unless otherwise specified. 60 , halo, -O - M + , -OR 70 , -SR 70 , -S - M + , -NR 80 R 80 , trihalomethyl, -CF3, -CN, -OCN, -SCN, -NO, -NO2, -N3, -SO2R 70 , -SO3 - M + , -SO3R 70 , -OSO2R 70 , -OSO3 - M + , -OSO3R 70 , -PO3-2 (M + )2, -P(O)(OR 70 )O - M + , -P(O)(OR 70 )2, -C(O)R 70 , -C(S)R 70 , -C(NR 70 )R 70 , -CO2 - M + , -CO2R 70 , -C(S)OR 70 , -C(O)NR 80 R 80 , -C(NR 70 )NR 80 R 80 , -OC(O)R 70 , -OC(S)R 70 , -OCO2 - M + , -OCO2R 70 , -OC(S)OR 70 , -NR 70 C(O)R 70 , -NR 70 C(S)R 70 , -NR 70 CO2 - M + , -NR 70 CO2R 70 , -NR 70 C(S)OR 70 , -NR 70 C(O)NR 80 R 80 , -NR 70 C(NR 70 )R 70 , and -NR 70 C(NR 70 )NR 80 R 80 where R 60 , R 70 , R 80 , and M + is as defined above, except that in the case of a substituted alkene or alkyne, the substituent may be -O - M + , -OR 70 , -SR 70 , or -S - M+ isn't it.

[0436] In addition to the groups disclosed for each individual term herein, the substituents of the hydrogen on the nitrogen atom in "substituted" heteroalkyl and cycloheteroalkyl groups are, unless otherwise specified, -R 60 , -O - M + , -OR 70 , -SR 70 , -S - M + , -NR 80 R 80 , trihalomethyl, -CF3, -CN, -NO, -NO2, -S(O)2R 70 , -S(O)2O - M + , -S(O)2OR 70 , -OS(O)2R 70 , -OS(O)2O - M + , -OS(O)2OR 70 , -P(O)(O - )2(M + )2, -P(O)(OR 70 )O - M + , -P(O)(OR 70 )(OR 70 ), -C(O)R 70 , -C(S)R 70 , -C(NR 70 )R 70 , -C(O)OR 70 , -C(S)OR 70 , -C(O)NR 80 R 80 , -C(NR 70 )NR 80 R 80 , -OC(O)R 70 , -OC(S)R 70 , -OC(O)OR 70 , -OC(S)OR 70 , -NR 70 C(O)R 70 , -NR 70 C(S)R 70 , -NR 70 C(O)OR 70 , -NR 70C(S)OR 70 , -NR 70 C(O)NR 80 R 80 , -NR 70 C(NR 70 )R 70 , and -NR 70 C(NR 70 )NR 80 R 80 where R 60 , R 70 , R 80 , and M + is as defined above.

[0437] Further to the disclosure herein, in certain embodiments, a substituted group has 1, 2, 3, or 4 substituents, 1, 2, or 3 substituents, 1 or 2 substituents, or 1 substituent.

[0438] Unless otherwise indicated, substituents not explicitly defined herein are named by naming the terminal portion of the functional group, followed by the adjacent functional group toward the point of attachment. For example, the substituent "arylalkyloxycarbonyl" refers to the group (aryl)-(alkyl)-OC(O)-.

[0439] With respect to any of the groups disclosed herein that contain one or more substituents, it is of course understood that such groups do not include substitutions or patterns that are sterically impractical and / or synthetically impractical. Furthermore, the subject compounds include all stereochemical isomers arising from the substitution of these compounds.

[0440] In certain embodiments, substituents may contribute to the optical isomerism and / or stereoisomerism of the compound. Salts, solvates, hydrates, and prodrug forms of the compound are also of interest. All such forms are encompassed by the present disclosure. Thus, the compounds described herein encompass salts, solvates, hydrates, prodrugs, and isomers of the compound, including pharmaceutically acceptable salts, solvates, hydrates, prodrugs, and isomers of the compound. In certain embodiments, the compound may be metabolized to a pharmaceutically active derivative.

[0441] Unless otherwise specified, a reference to an atom is intended to include isotopes of that atom. For example, a reference to H is 1 H, 2 H (i.e., D), and 3 is intended to encompass H (i.e., T), and reference to C is 12 C and all isotopes of carbon ( 13 C, etc.)

[0442] Unless otherwise indicated, the term "about" or "approximately" refers to an acceptable error of a particular value as determined by one of ordinary skill in the art, which depends in part on how the value is measured or determined. In certain embodiments, the term "about" or "approximately" means within 1, 2, or 3 standard deviations. In certain embodiments, the term "about" or "approximately" means within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.25%, 0.2%, 0.1%, or 0.05% of a given value or range. In certain embodiments requiring an integer, the term "about" means within ±10% of a given value or range, rounded up or down to the nearest integer.

[0443] In the description herein, if there is any discrepancy between a chemical name and a chemical structure, the chemical structure shall prevail.

[0444] Definitions of other terms and concepts are set forth throughout the detailed description.

[0445] ASGPR-binding compounds and conjugates are described in International Application No. PCT / US2021 / 012846, filed January 8, 2021, the disclosure of which is incorporated herein by reference in its entirety.

[0446] 5.8. Further Embodiments Further embodiments of the present disclosure are also described in the following clauses.

[0447] Clause 1. Formula (I): [ka] (In the formula, X is a moiety that binds to the cell surface asialoglycoprotein receptor (ASGPR), n is 1 to 500 (e.g., n is 1 to 20, 1 to 10, 1 to 6, or 1 to 5); L is a linker, Y is a biomolecule that specifically binds to the target protein A cell surface receptor-binding conjugate or a salt thereof.

[0448] Clause 2. Formula (V): [ka] (In the formula, n is 1 to 20, m is the average load number from 1 to 80, Ab is an antibody or antibody fragment that specifically binds to a target protein; Z is the residue resulting from the covalent attachment of the chemoselective linking group to a compatible group on Ab. 2. The conjugate according to clause 1, or a pharmaceutically acceptable salt thereof,

[0449] Clause 3. X is a moiety that binds to ASGPR and has formula (III-a) to (III-j): [ka] (In the formula, R 1 -OH, -OC(O)R, and [ka] (However, R is C 1~6 alkyl) is selected from R 2 are -NHCOCH3, -NHCOCF3, -NHCOCH2CF3, -OH, and [ka] is selected from R 3 is selected from -H, -OH, -CH3, -OCH3, and -OCH2CH=CH2 3. The conjugate according to clause 1 or 2, selected from:

[0450] Clause 4.X [ka] 4. The conjugate according to clause 3, wherein

[0451] Clause 5.X [ka] 4. The conjugate according to clause 3, wherein

[0452] Clause 6. The linker L has formula (IIa): [ka] (In the formula, Each L 1 ~L 7 are independently linking moieties and together provide a linear or branched linker between X and Y; a is 1 or 2, b, c, d, e, f, and g are each independently 0, 1, or 2; n is 1 to 6 (e.g., n is 1 to 5, or 2 to 6, or 1, 2, or 3) 6. The conjugate according to clauses 1 to 5,

[0453] Article 6a. If d is 0, then n is 1; When d is 1, n is 1 to 3; When d is 2, n is 1 to 6. A conjugate according to clause 6.

[0454] Article 7.-(L 1 ) a The conjugate of clause 6, wherein - comprises an optionally substituted aryl or heteroaryl linking moiety.

[0455] Article 8. Each L 1 Independently, [ka] (wherein v is 0 to 10, and z is 0 to 10) 7. The conjugate according to clause 6, selected from:

[0456] Article 9. Each L 2 independently, -C 1~6 -Alkylene-, -NHCO-C 1~6 -Alkylene-, -CONH-C 1~6 -Alkylene-, -O(CH2) p - and -(OCH2CH2) p - (wherein p is 1 to 10), Each L 3 Independently, [ka] , and -(OCH2CH2) q-wherein q is 1 to 10, u is 0 to 10, and w is 1 to 10.

[0457] If clause 10.n is 2 or more, at least one L 4 The conjugate of any one of clauses 6 to 9, wherein is present and is a branched chain linking moiety.

[0458] Article 11. Each L 4 Independently, -OCH2CH2-, [ka] (wherein each x and y independently represents an integer of 1 to 10) 11. The conjugate of any one of clauses 6 to 10, selected from:

[0459] Article 12. Each L 5 are independently, -NHCO-C 1~6 -Alkylene-, -CONH-C 1~6 -Alkylene-, -C 1~6 -alkylene-, [ka] , or -(OCH2CH2) r - and Each L 6 are independently, -NHCO-C 1~6 -Alkylene-, -CONH-C 1~6 -Alkylene-, -C 1~6 -Alkylene-, or -(OCH2CH2) s - and Each L 7 are independently, -NHCO-C 1~6 -Alkylene-, -CONH-C 1~6 -Alkylene-, -C 1~6 -Alkylene-, -(OCH2CH2) t - or -OCH2-, 12. The conjugate of any one of clauses 6 to 11, wherein r, s, and t are each independently 1 to 20.

[0460] The conjugate of any one of clauses 6 to 12, wherein clause 13.a is 1.

[0461] Clause 14. The conjugate of any one of clauses 6 to 13, wherein at least one of b, c, e, f, and g is not 0.

[0462] Clause 15. A conjugate according to any one of clauses 6 to 14, wherein at least one of b or c is not 0 and at least one of e, f and g is not 0.

[0463] Clause 16. The conjugate of any one of clauses 6 to 15, wherein a, b, and c are each independently 1 or 2.

[0464] Clause 17. The conjugate of any one of clauses 6-16, wherein the linker L is selected from any one of the structures in Tables 2-3.

[0465] Article 18. i) a conjugate resulting from the conjugation of any one of the compounds of the structures in the table of compounds described herein with a biomolecule, ii) a conjugate resulting from the conjugation of any one of the compounds of the structures in the table of compounds described herein with a polypeptide, or iii) A conjugate derived from the conjugation of any one of the compounds of the structure in the table of compounds described herein with an antibody or antibody fragment. 18. The conjugate of clause 1 or 17 selected from:

[0466] Clause 19. The conjugate of clause 18, wherein said antibody or antibody fragment is an IgG antibody.

[0467] Clause 20. The conjugate of clause 18, wherein said antibody or antibody fragment is a humanized antibody.

[0468] Clause 21. The conjugate of any one of clauses 18 to 20, wherein said antibody or antibody fragment specifically binds to a secreted or soluble protein.

[0469] Clause 22. The conjugate of any one of clauses 18 to 20, wherein said antibody or antibody fragment specifically binds to a cell surface receptor.

[0470] Clause 23. A method for internalizing a target protein into a cell containing an ASGPR cell surface receptor, comprising contacting said cell and a cell sample containing said target protein with an effective amount of a conjugate according to any one of clauses 1 to 22, wherein said conjugate specifically binds to said target protein and specifically binds to said cell surface receptor to promote cellular uptake of said target protein.

[0471] Clause 24. The method of Clause 23, wherein said target protein is a membrane-bound protein.

[0472] Clause 25. The method of clause 24, wherein said target protein is an extracellular protein.

[0473] Clause 26. The method of any one of clauses 23 to 25, wherein said compound or conjugate comprises an antibody or antibody fragment (Ab) that specifically binds to said target protein.

[0474] Clause 27. A method for reducing the level of a target protein in a biological system, comprising contacting said biological system with an effective amount of a conjugate according to any one of clauses 1 to 22, wherein said conjugate specifically binds to said target protein and specifically binds to an ASGPR cell surface receptor of cells in said biological system to promote cellular uptake and degradation of said target protein.

[0475] Clause 28. The method of claim 27, wherein said biological system is a human subject.

[0476] Clause 29. The method of claim 27, wherein said biological system is an in vitro cell sample.

[0477] Clause 30. The method of claim 28, wherein said target protein is a membrane-bound protein.

[0478] Clause 31. The method of claim 29, wherein said target protein is an extracellular protein.

[0479] Clause 32. A method for treating a disease or disorder associated with a target protein, comprising administering to a subject in need thereof an effective amount of a conjugate according to any one of clauses 1 to 22, wherein said conjugate specifically binds to said target protein.

[0480] Clause 33. The method of clause 32, wherein said disease or disorder is an inflammatory disease.

[0481] Clause 34. The method of clause 32, wherein said disease or disorder is an autoimmune disease.

[0482] Clause 35. The method of clause 32, wherein said disease or disorder is cancer.

[0483] Article 36. The following formula (I): [ka] (In the formula, X is the moiety that binds to the ASGPR cell surface receptor, L is of the following formula (IIa): [ka] (In the formula, Each L 1 independently, [ka] and Each L 2 are independent, -C 1~6 -Alkylene-, -NHCO-C 1~6 -Alkylene-, -CONH-C 1~6 -Alkylene-, -(OCH2) p - or -(OCH2CH2) p - and Each L 3 independently, [ka] , or -(OCH2CH2) q - and Each L 4 are independently -OCH2CH2-, [ka] and Each L 5 are independently -NHCO-C 1~6 -Alkylene-, -CONH-C 1~6 -Alkylene-, -C 1~6 -alkylene-, [ka] , or -(OCH2CH2) r - and Each L 6 are independently -NHCO-C 1~6 -Alkylene-, -CONH-C 1~6 -Alkylene-, -C 1~6 -Alkylene- or -(OCH2CH2) s - and Each L 7 are independently -NHCO-C 1~6 -Alkylene-, -CONH-C 1~6 -Alkylene-, -C 1~6 -Alkylene-, -(OCH2CH2) t - or -OCH2-, p, q, r, s, and t each independently represent an integer of 1 to 20; a is 1 or 2; b, c, d, e, f, and g each independently represent 0, 1, or 2; and u, v, w, x, y, and z each independently represent an integer of 1 to 10; n is an integer from 1 to 5, and when d is 0, n is 1, when d is 1, n is an integer from 1 to 3, and when d is 2, n is an integer from 1 to 5. is a linker for Y is [ka] (In the formula, [ka] represents the point of attachment to L, R is hydrogen or fluorine; each R' is independently hydrogen or halo; G is selected from -F, -Cl, -Br, -I, -O-mesyl, and -O-tosyl; J is -Cl, -Br, -I, -F, -OH, -ON-succinimide, -O-(4-nitrophenyl), -O-pentafluorophenyl, -O-tetrafluorophenyl, and -OC(O)-OR; J’ Selected from R J’ is -C1-C8 alkyl or -aryl) is a moiety selected from the group consisting of or a salt, single stereoisomer, mixture of stereoisomers, or isotopic form thereof.

[0484] The compound according to clause 36, wherein clause 37.a is 1.

[0485] Clause 38. The compound according to clause 36, wherein at least one of b, c, e, f and g is not 0.

[0486] Clause 39. The compound according to clause 36, wherein at least one of b or c is not 0 and at least one of e, f and g is not 0.

[0487] Clause 40. The compound according to clause 36, wherein a, b, and c are each independently 1 or 2.

[0488] Clause 41. Each X independently represents a group of formula (III-a) to (III-j): [ka] (In the formula, R 1 -OH, -OC(O)R, and [ka] (However, R is C 1~6 alkyl) is selected from R 2 are -NHCOCH3, -NHCOCF3, -NHCOCH2CF3, -OH, and [ka] is selected from R 3 is selected from -H, -OH, -CH3, -OCH3, and -OCH2CH=CH2 37. The compound according to clause 36, selected from:

[0489] Clause 42.X [ka] That is, the conjugate of Article 41.

[0490] Clause 43.X [ka] That is, the conjugate of Article 41.

[0491] Article 44. The following formula (IVa): [ka] (In the formula, X is the moiety that binds to the ASGPR cell surface receptor, L is of the following formula (IIa): [ka] (In the formula, Each L 1 independently, [ka] and Each L 2 are independent, -C 1~6 -Alkylene-, -NHCO-C 1~6 -Alkylene-, -CONH-C 1~6 -Alkylene-, -(OCH2) p - or -(OCH2CH2) p - and Each L 3 independently, [ka] , or -(OCH2CH2) q - and Each L 4 are independently -OCH2CH2-, [ka] and Each L 5 are independently -NHCO-C 1~6 -Alkylene-, -CONH-C 1~6 -Alkylene-, -C 1~6 -alkylene-, [ka] , or -(OCH2CH2) r - and Each L 6 are independently -NHCO-C 1~6 -Alkylene-, -CONH-C 1~6 -Alkylene-, -C 1~6 -Alkylene- or -(OCH2CH2)s - and Each L 7 are independently -NHCO-C 1~6 -Alkylene-, -CONH-C 1~6 -Alkylene-, C 1~6 -Alkylene-, -(OCH2CH2) t - or -OCH2-, p, q, r, s, and t each independently represent an integer of 1 to 20; a is 1 or 2; b, c, d, e, f, and g each independently represent an integer of 0, 1, or 2; and u, v, w, x, y, and z each independently represent an integer of 1 to 10. is a linker for n is an integer of 1 to 5, and when d is 0, n is 1, and when d is 1, n is an integer of 1 to 3, and when d is 2, n is an integer of 1 to 5; Z is [ka] (In the formula, [ka] represents the point of attachment to L, [ka] represents the point of attachment to P, X is CH2, NH, O, or S. is selected from the group consisting of P is a polypeptide or a pharmaceutically acceptable salt thereof.

[0492] Clause 45. A conjugate according to clause 44 wherein P comprises an antibody or an antigen-binding fragment of an antibody.

[0493] Article 46. The following formula (Va): [ka] (In the formula, X is the moiety that binds to the ASGPR cell surface receptor, L is of the following formula (IIa): [ka] (In the formula, Each L 1 independently, [ka] and Each L 2 are independent, -C 1~6 -Alkylene-, -NHCO-C 1~6 -Alkylene-, -CONH-C 1~6 -Alkylene-, -(OCH2) p - or -(OCH2CH2) p - and Each L 3 independently, [ka] , or -(OCH2CH2) q - and Each L 4 are independently -OCH2CH2-, [ka] and Each L 5 is -NHCO-C 1~6 -Alkylene-, -CONH-C 1~6 -Alkylene-, -C 1~6 -alkylene-, [ka] , or -(OCH2CH2) r - and Each L 6 is -NHCO-C 1~6 -Alkylene-, -CONH-C 1~6 -Alkylene-, -C 1~6 -Alkylene- or -(OCH2CH2) s - and Each L 7 is -NHCO-C 1~6 -Alkylene-, -CONH-C 1~6 -Alkylene-, C 1~6 -Alkylene-, -(OCH2CH2) t - or -OCH2-, p, q, r, s, and t are each independently an integer of 1 to 20; a is 1 or 2; b, c, d, e, f, and g are each independently 0, 1, or 2; and u, v, w, x, y, and z are each independently 1, 2, 3, 4, 5, or 6. is a linker for n is an integer of 1 to 5, and when d is 0, n is 1, and when d is 1, n is an integer of 1 to 3, and when d is 2, n is an integer of 1 to 5; m is an integer from 1 to 8; Z is [ka] (In the formula, [ka] represents the point of attachment to L, [ka] teeth [ka] represents the point of attachment to [ka] is an antibody) or a pharmaceutically acceptable salt thereof.

[0494] Clause 47. Each X independently represents a group of formula (III-a) to (III-j): [ka] (In the formula, R 1 -OH, -OC(O)R, and [ka] (However, R is C 1~6 alkyl) is selected from R 2 are -NHCOCH3, -NHCOCF3, -NHCOCH2CF3, -OH, and [ka] is selected from R 3 is selected from -H, -OH, -CH3, -OCH3, and -OCH2CH=CH2 47. The conjugate of any one of clauses 44 to 46, selected from:

[0495] Clause 48.X [ka] 48. The conjugate according to clause 47, wherein

[0496] Clause 49.X [ka] 48. The conjugate according to clause 47, wherein

[0497] Clause 50. A pharmaceutical composition comprising the conjugate or pharmaceutically acceptable salt according to any one of clauses 44 to 50 and a pharmaceutically acceptable carrier.

[0498] Clause 51. The pharmaceutical composition according to clause 50, wherein m is an integer from 4 to 8.

[0499] Clause 52. A pharmaceutical composition comprising the conjugate or pharmaceutically acceptable salt according to clause 51, wherein m is 4.

[0500] Clause 53. The conjugate of any one of clauses 44 to 49, wherein said antibody is an IgG antibody.

[0501] Clause 54. The conjugate of any one of clauses 44 to 49, wherein said antibody is a humanized antibody.

[0502] Clause 55. The conjugate of any one of clauses 44 to 49, wherein said antibody specifically binds to a secreted or soluble protein.

[0503] Clause 56. The conjugate of any one of clauses 44 to 49, wherein said antibody specifically binds to a cell surface receptor.

[0504] Clause 57. The conjugate of any one of clauses 44 to 49, wherein said antibody specifically binds to the Programmed Death Ligand-1 (PD-L1) protein.

[0505] Clause 58. The conjugate of any one of clauses 44 to 49, wherein said antibody specifically binds to vascular endothelial growth factor (VEGF) protein.

[0506] Clause 59. The conjugate of any one of clauses 44 to 49, wherein said antibody specifically binds to Fibroblast Growth Factor Receptor 2 (FGFR2) protein or Fibroblast Growth Factor Receptor 3 (FGFR3) protein.

[0507] Clause 60. The conjugate of any one of clauses 44 to 49, wherein said antibody is cetuximab.

[0508] Clause 61. The conjugate of any one of clauses 44 to 49, wherein said antibody is matuzumab.

[0509] Clause 62. The conjugate of any one of clauses 44 to 49, wherein said antibody is atezolizumab.

[0510] Clause 63. A method for treating a disease or disorder by administering to a subject in need thereof an effective amount of a conjugate or a pharmaceutically acceptable salt thereof according to any one of clauses 44 to 49, or a pharmaceutical composition according to any one of clauses 50 to 52.

[0511] Clause 64. The method of clause 63, wherein said disease or disorder is an inflammatory disease.

[0512] Clause 65. The method of clause 63, wherein said disease or disorder is an autoimmune disease.

[0513] Clause 66. The method of clause 63, wherein said disease or disorder is cancer.

[0514] Article 67. Formula (I): [ka] (In the formula, Y is the objective part, n is 1 to 500; L is a linker, X is a group represented by formula (Ia): [ka] (In the formula, R 1 -OH, -OC(O)R, -C(O)NHR, -Z 1 -*, and optionally substituted triazole, provided that R is optionally substituted C 1~6 alkyl or optionally substituted aryl; R 2 is -NHCOCH3, -NHCOCF3, -NHCOCH2CF3, -OH, optionally substituted triazole, and -Z 1 - Selected from *, R 3 -H, -OH, -CH3, -OCH3, -OCH2CH=CH, and -Z 1 - Selected from *, R 1 ~R3 One of them is -Z 1 -*, where "*" is Z for the linker (L). 1 represents the point of attachment of R 4 and R 5 are each independently selected from H and a promoiety (e.g., an ester promoiety); Z 1 is Z 11 , optionally replaced by Z 11 -heteroaryl, optionally substituted Z 11 - a linking moiety selected from aryl, optionally substituted heteroaryl, optionally substituted aryl, optionally substituted amide, optionally substituted sulfonamide, optionally substituted urea, and optionally substituted thiourea; Z 11 -O-, -S-, NR 21 - and -C(R 22 )2 is selected, Each R 21 are independently selected from H and optionally substituted (C1-C6) alkyl; Each R 22 are independently selected from H, halogen (e.g., F), and optionally substituted (C1-C6) alkyl. is the portion of the α-asialoglycoprotein receptor (ASGPR) that binds to the cell surface, i) n is 3 and R 1 is OH and R 2 is -NHCOCH3 and R 4 and R 5 is H and R 3 Z 1 If Z 1 is not O, ii) n is 2 or 3, and R 1 is OAc and R 2 is -NHCOCH3 and R 4 and R 5 is Ac and R 3 Z 1 If Z 1is not O, iii) n is 2 or 3, and R 1 is OBz and R 2 is -NHCOCH3 and R 4 and R 5 is Bz and R 3 Z 1 If Z 1 is not O, iv) n is 3 and R 1 is OH and R 2 is -NHCOCH3 and R 4 and R 5 is H and R 3 Z 1 and Z 1 is O, then L comprises a main chain of at least 16 consecutive atoms up to the branch point; and / or v) n is 3 and R 1 Z 1 (However, Z 1 is O) and R 4 and R 5 If is H, then R 3 is not -CH3) A cell surface ASGPR-binding compound, or a prodrug or salt thereof. [Example]

[0515] 6. Working Example The examples in this section are offered by way of illustration, not by way of limitation.

[0516] 6.1. Preparation of Compounds Below are illustrative schemes and examples of how the compounds described herein can be prepared and tested. While the examples may represent only some embodiments, it should be understood that the following examples are illustrative and not limiting. All substituents are as defined above unless otherwise indicated. Reagents and starting materials are readily available to one of ordinary skill in the art. Certain synthetic steps for each of the routes described may be combined in different forms or with steps from different schemes to prepare the compounds described herein.

[0517] Examples 1-54: Preparation of exemplary ASGPR ligand-linkers

[0518] Example 1: Synthesis of [(2R,3R,4R,5R,6R)-3,4-bis(acetyloxy)-6-(but-3-yn-1-yloxy)-5-acetamidooxan-2-yl]methyl acetate (Intermediate A) [ka]

[0519] To activated 4 Å molecular sieves (5.0 g) and [(2R,3R,4R,5R,6S)-3,4,6-tris(acetyloxy)-5-acetamidooxan-2-yl]methyl acetate (A-1) (5.0 g, 12.8 mmol) was added dichloromethane (50 mL) and stirred at room temperature for 5 minutes, followed by the addition of but-3-yn-1-ol (2.92 mL, 3.0 equiv., 38.5 mmol). The reaction mixture was stirred at room temperature for 10 minutes and then cooled to 0° C. Diethyl trifluoroborate (4.75 mL, 38.5 mmol) was added dropwise to the reaction mixture, which was again stirred at room temperature for 10 minutes and then refluxed at 51° C. for 5 hours. After confirming the completion of the reaction by TLC, the diethyl trifluoroborate was quenched by adding triethylamine (until a neutral pH was reached), filtered through a bed of Celite, and then concentrated on a rotary evaporator. The resulting viscous residue was purified by silica gel column purification using 60-75% ethyl acetate / dichloromethane as the eluent to give Intermediate A-2 as an off-white foam. Yield: 4.50 g, 87%; R f = 0.45 (7.5% methanol / dichloromethane); LC-MS m / z 400.0 [M+1] + ; 1 H NMR (400 MHz, CDCl3) δ 5.44 (d, J = 8.6 Hz, 1H), 5.35 (d, J = 7.0 Hz, 1H), 5.30 (dd, J = 11.2, 3.0 Hz, 1H), 4.79 (d, J = 8.2 Hz, 1H), 4.14 - 4.09 (m, 2H), 3.99 - 3.90 (m, 3H), 3.71 - 3.65 (m, 1H), 2.49 - 2.47 (m, 2H), 2.14 (s, 3H), 2.05 (s, 3H), 2.00 (s, 3H), 1.96 (s, 3H).

[0520] Intermediate A-2 (7.8 g, 17.5 mmol) was dissolved in methanol (50 mL) and cooled to 0 °C. To this solution, a 25% w / v solution of sodium methoxide in methanol (2.48 mL, 11.3 mmol) was added dropwise, and the reaction was maintained at room temperature for 3 h. After confirming the completion of the reaction by TLC, 1N HCl was added dropwise to quench the sodium methoxide. Methanol was evaporated, and the resulting residue was washed with diethyl ether (30 mL × 4). The resulting crude residue was purified by preparative HPLC (5-20% acetonitrile / water with 0.1% TFAH) to give Intermediate A as a white solid. Yield: 2.6 g, 84%; LC-MS m / z 274.0 [M+1] + ; 1 H NMR (400 MHz, D2O) δ 4.58 (d, J = 8.4 Hz, 1H), 3.97-3.86 (m, 3H), 3.82-3.73 (m, 5H), 2.49-2.44 (m, 2H), 2.04 (s, 3H).

[0521] Example 2: Synthesis of N-((2R,3R,4R,5R,6R)-6-((but-3-yn-1-yloxy)methyl)-2,4,5-trihydroxytetrahydro-2H-pyran-3-yl)acetamide (Intermediate B) [ka]

[0522] A solution of p-toluenesulfonyl chloride (1.1 equivalents) in dichloromethane is slowly added to a stirred solution of N-((2R,3R,4R,5R,6R)-2,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)acetamide (B-1) (1 equivalent) in dichloromethane at 0° C. The reaction mixture is warmed to room temperature and monitored by LC-MS until the formation of the tosylate ester of the desired primary alcohol is complete. Pyridine (3.5 equivalents) is added, followed by acetic anhydride (3.1 equivalents). The reaction mixture is stirred at room temperature and monitored by LC-MS until the formation of intermediate B-2 is complete, which is isolated by silica gel chromatography. To a stirred solution of but-3-yn-1-ol (1.1 equivalents) in tetrahydrofuran at 0° C. is added sodium hydride (1.1 equivalents) under 50° C. After stirring at 0°C for 10 minutes, a solution of intermediate B-2 (1 equivalent) in tetrahydrofuran is added. The resulting mixture is warmed to room temperature and monitored until LC-MS indicates complete formation of intermediate B-3, which is isolated by silica gel chromatography. To a stirred solution of intermediate B-3 (1 equivalent) in methanol at 0°C is added a solution of sodium methoxide in methanol (3 equivalents). The resulting mixture is stirred at 0°C until LC-MS indicates complete conversion to intermediate B, which is isolated by reverse-phase chromatography.

[0523] Example 3: Synthesis of trivalent GalNAc ligand A perfluorophenyl ester (compound I-107) [ka] [ka]

[0524] To a stirred solution of 2-(2-(2-azidoethoxy)ethoxy)ethan-1-ol (3A) (1 equivalent) and pyridine (1.2 equivalents) in dichloromethane is added a solution of p-toluenesulfonyl chloride (1.1 equivalents). The resulting mixture is stirred at room temperature and monitored until LC-MS indicates complete formation of compound 3B, which is isolated by silica gel chromatography. To a stirred mixture of tert-butyl (1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl)carbamate (1 equivalent) and compound 3B (3.3 equivalents) in THF at −78° C. is added sodium hydride. The cooling bath is removed, and the resulting mixture is stirred at room temperature until LC-MS indicates complete conversion to compound 3C, which is isolated by silica gel chromatography. To a stirred solution of compound 3C (1 equivalent) in dichloromethane at room temperature, a solution of HCl in diethyl ether (3 equivalents) is added. The resulting mixture is stirred at room temperature until LC-MS indicates complete conversion, and then the volatiles are removed on a rotary evaporator to give compound 3D. To a stirred solution of compound 3D (1 equivalent) in dichloromethane at room temperature, diisopropylethylamine (2 equivalents) is added. 3,3'-(ethane-1,2-diylbis(oxy))dipropionic acid bis(perfluorophenyl) (3E) (1.1 equivalents) is added, and the resulting mixture is stirred at room temperature until LC-MS indicates complete conversion to compound 3F, which is isolated by silica gel chromatography. Compound 3F (1 equivalent) and intermediate A (1 equivalent) are dissolved in DMSO at room temperature with stirring. Tetrakis(acetonitrile)copper(I) tetrafluoroborate (3 equivalents) is added and the resulting mixture is stirred at room temperature until LC-MS indicates complete conversion to compound I-107, which is purified by reverse-phase preparative HPLC and then lyophilized.

[0525] Example 4: Synthesis of trivalent GalNAc ligand B perfluorophenyl ester (compound I-108) [ka]

[0526] Compound 3F (1 equivalent) and intermediate B (1 equivalent) are dissolved in DMSO at room temperature under stirring. Tetrakis(acetonitrile)copper(I) tetrafluoroborate (3 equivalents) is added, and the resulting mixture is stirred at room temperature until LC-MS indicates complete conversion to compound I-108, which is purified by reverse-phase preparative HPLC and subsequently lyophilized.

[0527] Example 5: Synthesis of Divalent GalNAc Ligand A Perfluorophenyl Ester (Compound I-109) [ka]

[0528] Sodium hydride is added to a mixture of tert-butyl (1,3-dihydroxypropan-2-yl)carbamate (5A) (1 equivalent) and compound 3B (3.3 equivalents) in THF at −78° C. under stirring. The cooling bath is removed, and the resulting mixture is stirred at room temperature until LC-MS indicates complete conversion to compound 5B, which is isolated by silica gel chromatography. A solution of HCl in diethyl ether (3 equivalents) is added to a solution of compound 5B (1 equivalent) in dichloromethane under stirring at room temperature. The resulting mixture is stirred at room temperature until LC-MS indicates complete conversion, and then the volatiles are removed on a rotary evaporator to give compound 5C. Diisopropylethylamine (2 equivalents) is added to a solution of compound 5C (1 equivalent) in dichloromethane under stirring at room temperature. Bis(perfluorophenyl) 3,3'-(ethane-1,2-diylbis(oxy))dipropionate (compound 3E) (1.1 equivalents) is added, and the resulting mixture is stirred at room temperature until LC-MS indicates complete conversion to compound 5D, which is isolated by silica gel chromatography. Compound 5D (1 equivalent) and intermediate A (1 equivalent) are dissolved in DMSO at room temperature with stirring. Tetrakis(acetonitrile)copper(I) tetrafluoroborate (3 equivalents) is added, and the resulting mixture is stirred at room temperature until LC-MS indicates complete conversion to compound I-5, which is purified by reverse-phase preparative HPLC and then lyophilized.

[0529] Following the above synthesis, 41 mg of compound I-109 was obtained. LC-MS m / z 1336.7 [M+1] + ; 1HNMR (400 MHz, D2O) d 7.87 (s, 2H), 4.65-4.61 (m, 4H), 4.47 (d, J = 8.0 Hz, 2H), 4.23-4.11 (m, 2H), 4.01-3.91 (m, 10H), 3.88-3.82 (m, 10 H), 3.81 (s, 1H), 3.79-3.77 (m, 4H), 3.76-3.73 (m, 12 H), 3.72-3.68 (m, 14H). 3.63-3.55 (m, 6H), 3.09 (t, J = 6.0 Hz, 2H), 3.00 (t, J = 6.4 Hz, 4H), 1.88 (s, 6H).

[0530] Example 6: Synthesis of Divalent GalNAc Ligand B Perfluorophenyl Ester (Compound I-110) [ka]

[0531] Compound 5D (1 equivalent) and intermediate B (1 equivalent) are dissolved in DMSO at room temperature under stirring. Tetrakis(acetonitrile)copper(I) tetrafluoroborate (3 equivalents) is added, and the resulting mixture is stirred at room temperature until LC-MS indicates complete conversion to compound I-110, which is purified by reverse-phase preparative HPLC and subsequently lyophilized.

[0532] Example 7: 1-(4-(2-(((2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-1,2,3-triazol-1-yl)-12-(2-(2-(2-(4-(2-(((2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy- Synthesis of 6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethoxy)ethyl)-3,6,9,15,18,21-hexaoxa-12-azatetracosan-24-oic acid perfluorophenyl (divalent GalNAc ligand A perfluorophenyl ester, compound I-111) [ka]

[0533] 1-Azido-12-(2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethyl)-3,6,9,15,18,21-hexaoxa-12-azatetracosan-24-oic acid perfluorophenyl ester (7B, 1.0 equiv., 0.050 g, 0.063 mmol) and N-((2R,3R,4R,5R,6R)-2-(but-3-yn-1-yloxy)-4,5 To a solution of 2-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)acetamide (Intermediate A, 2.0 equiv., 0.034 g, 0.127 mmol) in dimethyl sulfoxide (2 mL) was added tetrakis(acetonitrile)copper(I) hexafluorophosphate (5.0 equiv., 0.118 g, 0.317 mmol), and the reaction mixture was stirred at room temperature for 1 h. After completion of the reaction, the reaction mixture was diluted with acetonitrile and purified by preparative HPLC (45-75% acetonitrile / water with 0.1% trifluoroacetic acid). Fractions containing the desired compound were combined and lyophilized to give 1-(4-(2-(((2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-1,2,3-triazol-1-yl)-12-(2-(2-(2-(4-(2-(((2R,3R,4R,5R,6R)-3- Acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)ethyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethoxy)ethyl)-3,6,9,15,18,21-hexaoxa-12-azatetracosan-24-oic acid perfluorophenyl ester (compound I-111) was obtained as a thick, syrup-like liquid. Yield: 0.044 g, 52.01%; LCMS m / z 1336.70 [M+1] + ; 1H-NMR (400 MHz, D2O) δ 7.86 (s, 2H), 4.65-4.61 (m, 4H), 4.47 (d, J = 8.0 Hz, 2H), 4.22-4.16 (m, 2H), 4.10-3.91 (m, 10H), 3.92-3.82 (m, 10H), 3.82-3.79 (m, 4H), 3.75-3.62 (m, 26H), 3.59-3.57 (m, 6H), 4.47 (t, J = 6.0 Hz, 2H), 2.99 (t, J = 5.6 Hz, 4H), 1.87 (s, 6H).

[0534] Example 8: Synthesis of GalNac Ligand A Perfluorophenyl Ester (Compound I-112) [ka]

[0535] To a solution of compound 3E (1.0 equiv., 0.50 g, 0.929 mmol) in tetrahydrofuran (5 mL, 10 vol) was added compound 8A (1.0 equiv., 0.203 g, 0.929 mmol) and N,N-diisopropylethylamine (2.0 equiv., 0.34 mL, 1.86 mmol). The reaction mixture was stirred at room temperature for 2 h. The reaction progress was monitored by LCMS. Upon completion, the reaction mixture was diluted with acetonitrile and purified by reverse-phase preparative HPLC (55–65% acetonitrile / water with 0.1% TFA). Fractions containing the desired product were combined and lyophilized to dryness to give perfluorophenyl 1-azido-13-oxo-3,6,9,16,19-pentaoxa-12-azadocosan-22-oic acid (compound 8B) as a colorless viscous liquid. Yield: 0.130 g, 23%; LCMS m / z 573.25 [M+1] + .

[0536] To a solution of compound 8B (1.0 equiv., 0.070 g, 0.122 mmol) in dimethyl sulfoxide (2 mL) was added intermediate A (1.0 equiv., 0.0334 g, 0.122 mmol). After stirring the reaction mixture for 5 minutes, tetrakis(acetonitrile)copper(I) hexafluorophosphate (2.5 equiv., 0.100 g, 0.306 mmol) was added. The reaction mixture was stirred at room temperature for 1 hour. The reaction progress was monitored by LCMS. After completion of the reaction, the reaction mixture was diluted with acetonitrile and purified by preparative HPLC (35-55% acetonitrile / water with 0.1% TFA). Fractions containing the desired product were combined and lyophilized to dryness to give compound I-112 as a colorless viscous liquid. Yield: 0.015 g, 14.5%; LCMS m / z 846.33 [M+1] + ; 1 H NMR (400 MHz, D2O) 7.83 (s, 1H), 4.60-4.58 (m, 2H), 4.43 (d, J = 8.4 Hz, 1H), 4.17-4.13 (m, 1H), 3.97-3.90 (m, 5H), 3.88-3.72 (m, 6H), 3.70-3.49 (m, 16H), 3.37-3.34 (m, 2H), 3.05 (t, J = 6.0 Hz, 2H), 2.96 (t, J = 6.0 Hz, 2H), 2.50 (t, J = 6.0 Hz, 2H), 1.84 (s, 3H).

[0537] Example 9 Synthesis of perfluorophenyl 1-(4-(2-(((2R,3R,4R,5R,6R)-5-acetamido-3,4,6-trihydroxytetrahydro-2H-pyran-2-yl)methoxy)ethyl)-1H-1,2,3-triazol-1-yl)-13-oxo-3,6,9,16,19-pentaoxa-12-azadocosan-22-oic acid (Compound I-113) [ka]

[0538] Compound I-113 is prepared by adapting the synthetic procedure for I-112, substituting intermediate A for intermediate B.

[0539] Example 10: Synthesis of Compounds I-114 to I-118 [ka]

[0540] To a solution of 1-azido-3,6,9,12,15,18-hexaoxahenicosan-21-oic acid perfluorophenyl (10A) (1.0 equiv., 0.0998 g, 0.183 mmol) in dimethyl sulfoxide (1 mL) was added Intermediate A (1.0 equiv., 0.050 g, 0.183 mmol) and stirred for 5 min. Then, tetrakis(acetonitrile)copper(I) hexafluorophosphate (2.5 equiv., 0.170 g, 0.457 mmol) was added. (mmol) was added and the reaction mixture was stirred at room temperature for 1 hour. The progress of the reaction was monitored by LC-MS. After the reaction was complete, the reaction mixture was diluted with acetonitrile and purified by preparative HPLC (30-45% acetonitrile / water with 0.1% acetic acid). Fractions containing the desired product were combined and lyophilized to dryness to give compound I-114 as an off-white solid. Yield: 0.022 g, 14.1%; LC-MS m / z 819.24 [M+1] + ; 1 H NMR (400 MHz, D2O) δ 7.81 (s, 1H), 4.57-4.55 (m, 2H), 4.40 (d, J = 19.2 Hz, 1H), 4.16-4.11 (m, 1H), 3.94-3.85 (m, 6H), 3.80-3.73 (m, 3H), 3.71-3.59 (m, 22H), 3.04 (t, J = 5.6 Hz, 2H), 2.94 (t, J = 6.0 Hz, 2H), 1.81 (s, 3H).

[0541] Compounds I-115 to I-118 were prepared by adapting the procedure for compound I-114. The structures and corresponding data for each of compounds I-115 to I-118 are shown below. [Table 33]

[0542] Example 11: Synthesis of Intermediate C [ka]

[0543] Dichloromethane is added to activated 4 Å molecular sieves and [(2R,3R,4R,5R,6S)-3,4,6-tris(acetyloxy)-5-acetamidooxan-2-yl]methyl acetate (C-1) (1 equivalent). But-3-yn-1-amine (3 equivalents) is added to the reaction solution. After cooling the reaction mixture to 0 °C, diethyl trifluoroborate (2 equivalents) is added. The reaction is stirred at room temperature and then heated to reflux for 16 h. Aqueous NaHCO3 is added to quench the diethyl trifluoroborate, and the DCM layer is partitioned and dried over MgSO4. The solution is filtered and concentrated on a rotary evaporator. Silica gel column purification using 60-75% ethyl acetate / dichloromethane as the eluent affords intermediate C-2.

[0544] Intermediate C-2 (1 equivalent) is dissolved in methanol and cooled to 0°C. To this solution, a 25% w / v solution of sodium methoxide in methanol (10 equivalents) is added dropwise. The reaction is maintained at room temperature for 3 hours. After the reaction is complete, 1N HCl is added dropwise to quench the sodium methoxide. The methanol is evaporated, and the resulting residue is washed with diethyl ether. The resulting crude residue is purified by preparative HPLC (5-20% acetonitrile / water with 0.1% TFAH) to give Intermediate C.

[0545] Example 12: Synthesis of Compound I-120 [ka]

[0546] Compound 5D (1 equivalent) and intermediate C (1 equivalent) are dissolved in DMSO at room temperature under stirring. Tetrakis(acetonitrile)copper(I) tetrafluoroborate (3 equivalents) is added, and the resulting mixture is stirred at room temperature until LC-MS indicates complete conversion to compound I-120, which is purified by reverse-phase preparative HPLC and subsequently lyophilized.

[0547] Example 13: Synthesis of Compound I-146 [ka]

[0548] Compound 17B was synthesized using the procedure described for compound 8B, substituting compound 17A for compound 8A. Compound I-146 was synthesized using the procedure described for compound I-8, substituting compound 17B for compound 8B (32 mg). LC-MS m / z 978.3 [M+1] + .

[0549] Example 14: Synthesis of Compound I-122 [ka] [ka]

[0550] To a solution of compound A-1 (1.0 equiv., 5.05 g, 13.0 mmol) and benzyl N-[3-(5-hydroxypentanamido)propyl]carbamate (compound 18A) (1.0 equiv., 4.00 g, 13.0 mmol) in dichloromethane (50.0 mL) was added trimethylsilyl trifluoromethanesulfonate (1.1 equiv., 2.52 mL, 14.3 mmol) at room temperature. The reaction mixture was stirred at 40 °C for 5 h. After completion of the reaction, the reaction mixture was quenched with saturated sodium bicarbonate solution and extracted with dichloromethane. The organic layer was dried over sodium sulfate, filtered, and concentrated under high vacuum to give the crude product. The crude product was purified by reverse-phase chromatography using 0-30% acetonitrile / water to give compound 18B as a yellow viscous liquid. Yield: (5.80g, 70.12%); LCMS m / z 638.2 [M+1] + .

[0551] To a solution of compound 18B (1.0 equiv., 4.80 g, 7.53 mmol) in methanol (40.0 mL), 10% palladium on carbon (1.60 g) was added and stirred under a hydrogen atmosphere at room temperature for 4 hours. After the reaction was completed, the reaction mixture was filtered through a syringe filter, and the filtrate was concentrated and dried to give the crude product. The crude product was triturated with diethyl ether to give compound 18C as a pale yellow viscous liquid. Yield: (3.4 g, 80.73%); LCMS m / z 504.37 [M+1] + .

[0552] A solution of 2,3,4,5,6-pentafluorophenyl 3-(2-{[(benzyloxy)carbonyl]amino}-3-[3-oxo-3-(2,3,4,5,6-pentafluorophenoxy)propoxy]-2-{[3-oxo-3-(2,3,4,5,6-pentafluorophenoxy)propoxy]methyl}propoxy)propanoate (18D) (1.0 equiv., 1.20 g, 1.24 mmol) and compound 18C (3.0 equiv., 1.87 g, 3.71 mmol) in N,N-dimethylformamide (30.0 mL) was stirred at room temperature for 1 hour. After completion of the reaction, the reaction mixture was concentrated and dried to give the crude product. The crude product was purified by flash column chromatography using 20% ​​methanol / dichloromethane to give compound 18E as a pale yellow viscous liquid. Yield: (1.60g; 67.05%); LCMS m / z 1926.78 [M-1] - .

[0553] To a solution of compound 18E (1.0 equiv., 1.60 g, 0.830 mmol) in methanol (20 mL) and acetic acid (1.0 mL) was added 10% palladium on carbon (250 mg). The reaction mixture was stirred under a hydrogen atmosphere at room temperature for 16 hours. After the reaction was completed, the reaction mixture was filtered through a celite bed, and the filtrate was concentrated and dried to give compound 18F as a pale yellow viscous liquid. Yield: 1.45 g (crude); LCMS m / z 1794.05 [M+1] + .

[0554] To a solution of compound 18F (1.0 equiv., 1.45 g, 0.808 mmol) in methanol (10 mL) was added 25% sodium methanolate solution (8.0 equiv., 1.45 mL, 6.47 mmol) at 0° C. The reaction mixture was stirred at room temperature for 1 hour. After the reaction was completed, the reaction mixture was concentrated and dried to give the crude product. This crude product was diluted with acetonitrile and purified by preparative HPLC (30% acetonitrile / water with 0.1% TFA). Fractions containing the desired product were combined and lyophilized to dryness to give compound 18G as an off-white semi-solid. Yield: (0.20 g, 17.4%); LCMS m / z 1415.77 [M+1]+ .

[0555] To a solution of compound 18G (1.0 equiv., 0.090 g, 0.0636 mmol) in dimethyl sulfoxide (1.00 mL), compound 3E (1.0 equiv., 0.030 g, 0.0636 mmol) was added and stirred at room temperature for 16 h. After completion of the reaction, the reaction mixture was diluted with acetonitrile and purified by preparative HPLC (42% acetonitrile / water with 0.1% acetic acid (0–13 min)). Fractions containing the desired product were combined and lyophilized to dryness to give compound I-122 as an off-white solid. Yield: 0.004 g, 3.55%; LC-MS m / z 1769.93 [M+1] + ; 1 H NMR (400 MHz, DMSO-d6) δ 7.84 (bs, 3H), 7.73 (bs, 3H), 7.63 (d, J = 9.2 Hz, 3H), 7.13 (s, 1H), 4.58-4.54 (m, 4H), 4.47 (bs, 3H), 4.22 (d, J = 8.8 Hz, 3H), 3.77-3.67 (m, 12H), 3.53-3.52 (m, 30H), 3.32-3.27 (m, 4H), 3.02 (bs, 14H), 2.29 (t, J = 6.0 Hz, 6H), 2.05 (t, J = 7.2 Hz, 6H), 1.79 (s, 9H), 1.50-1.41 (m, 18H).

[0556] Example 15 Synthesis of N-[1,3-bis(2-{[3-(5-{[(2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)])oxan-2-yl]oxy}pentanamido)propyl]carbamoyl}ethoxy)-2-[(2-{[3-(5-{[(2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}pentanamido)propyl]carbamoyl}ethoxy)methyl]propan-2-yl]-12-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)dodecanamide (Compound I-123) [ka]

[0557] To a solution of 12-aminododecanoic acid (19A) (2.00 g, 9.29 mmol) in acetic acid (15.00 ml) was added 2,5-dihydrofuran-2,5-dione (1.09 g, 11.1 mmol), and the reaction mixture was refluxed at 120° C. for 16 hours. After the reaction was completed, the reaction mixture was concentrated under vacuum to give the crude compound, which was purified by flash column chromatography using silica gel and 5% methanol / dichloromethane as the eluent to give compound 19B as an off-white solid. Yield: 1.60 g (57.17%); LCMS m / z 294.3 [M-1] - .

[0558] To a solution of compound 19B (0.300 g, 1.02 mmol) in tetrahydrofuran (15.00 mL) at 0 °C, pentafluorophenol (168 mg, 0.914 mmol) and diisopropylmethanediimine (0.192 mL, 1.22 mmol) were added. The reaction mixture was then stirred at room temperature for 1 hour. After the reaction was complete, the reaction mixture was concentrated to give the crude product, which was purified by flash column chromatography using silica gel and 5% to 7% ethyl acetate / hexane as the eluent to give compound 19C as an off-white solid. Yield: 0.250 g (53.34%), LCMS m / z 479.0 [M+18]. + .

[0559] To a solution of compound 18G (0.060 g, 0.04 mmol) in dimethyl sulfoxide (1.0 mL), N,N-diisopropylethylamine (0.015 mL, 0.084 mmol) and compound 19C (0.019 g, 0.04 mmol) were added, and the reaction mixture was stirred at room temperature for 16 h. After completion of the reaction, the reaction mixture was diluted with acetonitrile and purified by preparative HPLC (25-45% acetonitrile / water with 0.1% TFA). Fractions containing the desired product were combined and lyophilized to dryness to give compound I-123 as an off-white solid. Yield: 0.0035 g, 4.88%; LC-MS m / z 1692.93 [M+1] + ; 1 H NMR (400 MHz, DMSO-d6) δ 7.83 (t, J = 5.2 Hz, 3H), 7.73 (t, J = 6.0 Hz, 3H), 7.61 (d, J = 9.6 Hz, 3H), 6.98 (s, 3H), 4.57-4.53 (m, 7H), 4.22 (d, J = 8.4 Hz, 3H), 3.72-3.63 (m, 9H), 3.55-3.51 (m, 20H), 3.37-3.27 (m, 10H), 3.04-3.01 (m, 12H), 2.29 (t, J = 6.4 Hz, 6H), 2.05 (t, J = 6.8 Hz, 6H), 1.81 (bs, 8H), 1.51-1.41 (m, 24H), 1.21 (s, 14H).

[0560] Example 16: Synthesis of Compounds I-124 and I-132 [ka]

[0561] To a solution of dodecanedioic acid (20A) (1.00 g, 4.34 mmol) in ethyl acetate (10.00 mL) at 0° C., pentafluorophenol (1.60 g, 8.68 mmol) and diisopropylmethanediimine (1.91 mL, 13.0 mmol) were added, and the reaction mixture was stirred at room temperature for 1 hour. After the reaction was completed, the reaction mixture was filtered through a Celite bed, and the filtrate was concentrated under reduced pressure to obtain the crude compound. The obtained crude compound was purified by flash column chromatography on a silica gel column using 5% ethyl acetate / hexane as the eluent to obtain compound 20B as an off-white solid. Yield: 1.00 g (40.95%); LCMS m / z 580.39 [M+18] + .

[0562] To a solution of compound 18G (45.0 mg, 0.031 mmol) in dimethyl sulfoxide (1.0 mL) were added N,N-diisopropylethylamine (0.016 mL, 0.093 mmol) and compound 20B (17.9 mg, 0.031 mmol). The reaction mixture was stirred at room temperature for 2 hours. After completion of the reaction, the reaction mixture was purified by preparative HPLC (40-60% acetonitrile / water with 0.1% trifluoroacetic acid). Fractions containing the desired product were combined and lyophilized to dryness to give compound I-124 as an off-white solid. Yield: 0.006 g (10.52%); LCMS m / z 1793.94 [M+1] + , 897.99 [M / 2+1] + . 1H NMR (400 MHz, DMSO-d6) δ 7.83 (t, J = 5.6 Hz, 3H), 7.73 (t, J = 5.2 Hz, 3H), 7.60 (d, J = 9.2 Hz, 3H), 6.99 (s, 1H), 4.57-4.47 (m, 6H), 4.46 (d, J = 4.4 Hz, 3H), 4.21 (d, J = 8.4 Hz, 3H), 3.70-3.63 (m, 9H), 3.55-3.49 (m, 21H), 3.32-3.28 (m, 4H), 3.02 (t, J = 5.6 Hz, 12H), 2.76 (t, J = 5.6 Hz, 2H), 2.27 (t, J = 6.4 Hz, 6H), 2.03 (t, J = 7.2 Hz, 8H), 1.79 (s, 9H), 1.70-1.67 (m, 2H), 1.52-1.41 (m, 20H), 1.23 (bs, 14H).

[0563] Compound I-132 is synthesized by adapting the procedure for compound I-124. The structure of compound I-132 is shown below. [ka]

[0564] Example 17: Synthesis of Compound I-163 [ka]

[0565] To a mixture of compound I-124 (1 equiv., 17.2 mg, 0.00959 mmol) and 1-(2-aminoethyl)pyrrole-2,5-dione hydrochloride (1.1 equiv., 1.86 mg, 0.00105 mmol) in NMP (0.5 mL) was added DIEA (3 equiv., 5 μL, 0.0288 mmol). The mixture was stirred at room temperature for 10 min, and acetic acid (4 μL) was added. The mixture was purified by preparative HPLC (10-40% MeCN / water with 0.1% TFA) to give compound I-163 (10.1 mg, 64% yield) as a white solid (purity: 99%). LCMS m / z 1751.0 [M+H] + .

[0566] Example 18: Synthesis of Compounds I-125 and I-145 [ka]

[0567] To a solution of compound 18G (1.0 equiv., 0.10 g, 0.070 mmol) in dimethyl sulfoxide (1.00 mL), ethylbis(propan-2-yl)amine (3.0 equiv., 39.1 μL, 0.212 mmol) and bis(2,3,4,5,6-pentafluorophenyl) 4,7,10,13,16,19,22,25,28-nonaoxahentriacontanedioate (21A) (1.0 equiv., 0.0598 g, 0.070 mmol) were added and stirred at room temperature for 16 h. After completion of the reaction, the reaction mixture was diluted with acetonitrile and purified by preparative HPLC (50% acetonitrile / water with 0.1% acetic acid (0–10 min)). Fractions containing the desired product were combined and lyophilized to dryness to give compound I-125 as an off-white solid. Yield: 0.006g, 4.09%; LC-MS m / z 1039.74 [M / 2+1] + ; 1HNMR (400 MHz, D2O) δ 4.45 (d, J = 8.4 Hz, 3H), 3.96-3.83 (m, 11H), 3.80-3.58 (m, 61H), 3.24-3.19 (m, 12H), 3.10 (t, J = 5.6 Hz, 2H), 2.52-2.47 (m, 8H), 2.27 (t, J = 6.0 Hz, 6H), 2.02 (s, 9H), 1.75-1.70 (m, 6H), 1.58-1.50 (m, 12H), 1.35-1.34 (m, 1H).

[0568] Compound I-145 was synthesized by adapting the synthetic procedure of compound I-125. The structure and data of compound I-145 are shown below. [ka]

[0569] MS(ESI) m / z 1858 [M+1] + , 729 [M / 2+1] + .

[0570] 1 H NMR (400 MHz, DMSO-d6) δ 7.83 (t, J = 5.6 Hz, 3H), 7.35 (t, J = 5.2 Hz, 3H), 7.61 (d, J = 8.8 Hz, 3H), 7.13 (s, 1H), 4.59-4.54 (m, 6H), 4.46 (d, J = 4.4 Hz, 3H), 4.21 (d, J = 8.4 Hz, 3H), 3.76-3.70 (m, 2H), 3.67-3.63 (m, 10H), 3.55-3.46 (m, 34H), 3.14 (s, 2H), 3.32-3.28 (m, 2H), 3.02 (t, J = 6 Hz, 16H), 2.27 (t, J = 6 Hz, 6H), 2.03 (t, J = 7.2 Hz, 6H), 1.79 (s, 9H), 1.51-139 (m, 20H).

[0571] Example 19: Synthesis of Compounds I-159 and I-170 Compound I-159 was synthesized according to the procedure for compound I-125, substituting intermediate 22-B for intermediate 21-A. [ka]

[0572] The structure and data of compound I-159 are shown below. [ka]

[0573] LCMS m / z 1698.9 [M+H] + .

[0574] Compound I-170 was synthesized according to the procedures for compounds I-125 and I-159, substituting 3-[2-[2-[2-[2-[2-[2-(2,5-dioxopyrrol-1-yl)ethoxy]ethoxy]ethoxy]propanoic acid(2,3,4,5,6-pentafluorophenyl) for intermediate 22-B (3-[2-[2-[2-(2,5-dioxopyrrol-1-yl)ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid(2,3,4,5,6-pentafluorophenyl).

[0575] The structure and data of compound I-170 are shown below. [ka]

[0576] LCMS m / z 1831.7 [M+H] + .

[0577] Example 20: Synthesis of Compounds I-127 and I-129 [ka]

[0578] To a solution of bis(perfluorophenyl) 3,3'-((2-((3-oxo-3-(perfluorophenoxy)propoxy)methyl)-2-(pent-4-ynamido)propane-1,3-diyl)bis(oxy))dipropionic acid ester (23A) (1.0 equiv., 0.500 g, 0.54 mmol) and compound 18C (4.0 equiv., 1.3 g, 2.16 mmol) in N,N-dimethylformamide (10 mL) was added N,N-diisopropylethylamine (6.0 equiv., 0.59 mL, 3.24 mmol), and the reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction, the reaction mixture was concentrated and dried to give compound 23B as a pale brown viscous liquid. Yield: 3.0 g (crude), LCMS m / z 937.4 [M+2H] ++ .

[0579] To a solution of compound 23B (1.0 equiv., 3.0 g, 1.60 mmol) in methanol (10 mL) was added sodium methoxide (25% in methanol) (10.0 equiv., 3.92 mL, 16.0 mmol), and the reaction mixture was stirred at room temperature for 1 hour. The reaction was monitored by electroluminescence (ELSD). After completion of the reaction, the reaction mixture was neutralized with Dowex 50WX8 hydrogen form (200-400 mesh) and filtered. The filtrate was concentrated to give the crude product, which was diluted with acetonitrile and purified by preparative HPLC (13-25% acetonitrile / water). Fractions containing the desired product were combined and lyophilized to dryness to give compound 23C as an off-white solid. Yield: 0.380 g, 15.45%; LCMS m / z 748.35 [M+2H] ++ .

[0580] To a solution of compound 23C (1.0 equiv., 0.040 g, 0.026 mmol) in dimethyl sulfoxide (1.0 mL), compound 13A (1.2 equiv., 0.010 g, 0.032 mmol) was added and stirred for 5 minutes. Tetrakis(acetonitrile)copper(I) hexafluorophosphate (2.8 equiv., 0.027 g, 0.074 mmol) was then added, and the reaction mixture was stirred at room temperature for 15 minutes. After completion of the reaction, the reaction mixture was diluted with acetonitrile and purified by preparative HPLC (20-45% acetonitrile / water with 0.1% TFA). Fractions containing the desired product were combined and lyophilized to dryness to give compound I-127 as an off-white solid. Yield: 0.008 g, 16.6%; LCMS m / z 911.31 [M+2H] ++ ; 1 H NMR (400 MHz, D2O) δ 7.71 (s, 1H), 4.57-4.54 (m, 3H), 4.39 (d, J = 8.4 Hz, 4H), 3.94 (t, J = 8.4 Hz, 2H), 3.91-3.79 (m, 10H), 3.76-3.72 (m, 5H), 3.69-3.67 (m, 2H), 3.65-3.63 (m, 10H), 3.58 (bs, 5H), 3.55-3.52 (m, 4H), 3.18-3.13 (m, 12H), 2.95 (t, J = 5.2 Hz, 2H), 2.81 (t, J = 6.8 Hz, 2H), 2.49-2.46 (m, 2H), 2.44-2.41 (m, 6H), 2.19-2.17 (m, 6H), 1.98 (s, 9H), 1.69-1.62 (m, 6H), 1.60-1.49 (m, 12H).

[0581] Compound I-129 was synthesized by adapting the synthetic procedure for compound I-127. The structure and data for compound I-129 are shown below. [ka]

[0582] LCMS m / z 1065.25 [M+2H]++ .

[0583] 1 H NMR (400 MHz, D2O) δ 7.81 (s, 1H), 4.55 (bs, 2H), 4.39 (d, J = 8.4 Hz, 3H), 3.89-3.82 (m, 12H), 3.78-3.74 (m, 5H), 3.71-3.58 (m, 51H), 3.19-3.14 (m, 12H), 3.04 (t, J = 5.2 Hz, 2H), 2.91 (t, J = 7.2 Hz, 2H), 2.56 (t, J = 7.6 Hz, 2H), 2.42 (bs, 6H), 2.21-2.10 (m, 6H), 1.98 (s, 9H), 1.66 (t, J = 6.8Hz, 6H), 1.53 (BS, 12H).

[0584] Example 21 Synthesis of perfluorophenyl 1-(((2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-16-((3-((3-(5-(((2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)pentanamido)propyl)amino)-3-oxopropoxy)methyl)-5,11,18-trioxo-14-oxa-6,10,17-triazanonacosan-29-oic acid (Compound I-144) [ka] [ka]

[0585] To a stirred solution of 2-aminopropane-1,3-diol (21-1, 1.00 g, 11.0 mmol) in dimethyl sulfoxide (27 mL) was added aqueous sodium hydroxide (3.62 mmol / L, 4 mL) at 0 °C. tert-Butyl acrylate (3.52 g, 27.4 mmol) dissolved in dimethyl sulfoxide (4.0 mL) was slowly added, and the reaction mixture was stirred at room temperature for 4 hours. Water was added to the reaction mixture, followed by extraction with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the crude product, which was purified by silica gel column chromatography using 30% ethyl acetate / hexane to give di-tert-butyl 3,3'-((2-aminopropane-1,3-diyl)bis(oxy))dipropionate (21-2). Yield: 1.20 g, 32%. ESI-MS m / z: 348.0 (M+H) + .

[0586] To a stirred solution of di-tert-butyl 3,3'-((2-aminopropane-1,3-diyl)bis(oxy))dipropionate (21-2, 1.20 g, 3.45 mmol) in 1,4-dioxane (20.0 mL) was added sodium carbonate (0.54 g, 5.18 mmol) dissolved in water (4.0 mL). The reaction mixture was then placed in an ice bath, and benzyl chloroformate (0.74 mL, 5.18 mmol) was added. After 10 minutes, the ice bath was removed and the mixture was stirred at room temperature for an additional 3 hours. The reaction mixture was diluted with water and extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the crude product, which was purified by flash column chromatography using 4-7% methanol / dichloromethane to give di-tert-butyl 3,3'-((2-(((benzyloxy)carbonyl)amino)propane-1,3-diyl)bis(oxy))dipropionate (21-3). Yield: 1.0 g, 60.12%; LCMS m / z 482.0 [M+1] + .

[0587] A solution of di-tert-butyl 3,3'-((2-(((benzyloxy)carbonyl)amino)propane-1,3-diyl)bis(oxy))dipropionate (21-3, 1.00 g, 11.0 mmol) in formic acid (5.0 mL) was stirred at room temperature for 48 hours. The reaction mixture was concentrated under reduced pressure to give crude 3,3'-((2-(((benzyloxy)carbonyl)amino)propane-1,3-diyl)bis(oxy))dipropionic acid (21-4). The crude product obtained was used directly in the next reaction. Yield: 0.380 g, 49.29%. 1 H NMR (400 MHz, DMSO-d6) δ 7.38-7.29 (m, 5H), 7.14 (d, J= 8 Hz, 1H), 5.01 (s, 1H), 3.72 (t, J= 6.8 Hz, 4H) 3.34 (d, J= 6 Hz., 4H), 2.42(t, J = 6 Hz 4H).

[0588] To a stirred solution of 3,3'-((2-(((benzyloxy)carbonyl)amino)propane-1,3-diyl)bis(oxy))dipropionic acid (21-4, 0.750 g, 2.03 mmol) in ethyl acetate (10.0 mL) was added a solution of diisopropylmethanediimine (0.894 mL, 3.0 equiv., 6.09 mmol) and pentafluorophenol (5.86 g, 5.0 equiv., 31.8 mmol) in ethyl acetate (10.0 mL), and the reaction mixture was stirred at room temperature for 3 hours. After completion of the reaction, the reaction mixture was filtered through a pad of Celite, and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by column chromatography using silica gel (100-200 mesh) eluting with 20-30% ethyl acetate / hexane to give bis(perfluorophenyl) 3,3'-((2-(((benzyloxy)carbonyl)amino)propane-1,3-diyl)bis(oxy))dipropionate (21-5) as an off-white solid. Yield: 0.69 g, 47.96%. 1H NMR (400 MHz, DMSO-d6) δ 7.32 (s, 5H), 7.13 (d, J = 8 Hz, 1H), 4.99 (s, 2H), 4.10-3.94 (m, 5H ) 3.42(t, J = 6 Hz., 4H), 3.00 (t, J = 5.6 Hz 4H).

[0589] To a stirred solution of bis(perfluorophenyl) 3,3'-((2-(((benzyloxy)carbonyl)amino)propane-1,3-diyl)bis(oxy))dipropionate (21-5, 0.680 g, 1.0 equiv., 0.969 mmol) and (2R,3R,4R,5R,6R)-5-acetamido-2-(acetoxymethyl)-6-((5-((3-aminopropyl)amino)-5-oxopentyl)oxy)tetrahydro-2H-pyran-3,4-diyl diacetate (21-6, 1.95 g, 4.0 equiv., 3.88 mmol) in acetonitrile, DIPEA was added, and the reaction mixture was stirred at room temperature for 2 h. The solvent was evaporated under reduced pressure to give the crude product, which was purified by flash chromatography using 10% methanol / dichloromethane to give (2R,2'R,3R,3'R,4R,4'R,5R,5'R,6R,6'R)-((16-(((benzyloxy)carbonyl)amino)-5,11,21,27-tetraoxo-14,18-dioxa-6,10,22,26-tetraazahentriacontane-1,31-diyl)bis(oxy))bis(5-acetamido-2-(acetoxymethyl)tetrahydro-2H-pyran-6,3,4-triyl)tetraacetic acid (21-7) as a colorless liquid. Yield: 1.10 g, 62.94%; LCMS m / z 670.8 [M / 2+1] + .

[0590] To a stirred solution of ((2R,2'R,3R,3'R,4R,4'R,5R,5'R,6R,6'R)-((16-(((benzyloxy)carbonyl)amino)-5,11,21,27-tetraoxo-14,18-dioxa-6,10,22,26-tetraazahentriacontane-1,31-diyl)bis(oxy))bis(5-acetamido-2-(acetoxymethyl)tetrahydro-2H-pyran-6,3,4-triyl)tetraacetic acid (21-7, 1.10 g, 0.821 mmol) in methanol (10.0 mL) was added acetic acid (0.0469 mL, 0.821 mmol) and 10% palladium on carbon (0.30 0 g) was added, and the reaction mixture was stirred under a hydrogen gas atmosphere at room temperature for 2 hours. The reaction mixture was then passed through a Celite pad, and the filtrate was evaporated under reduced pressure to give crude (2R,2'R,3R,3'R,4R,4'R,5R,5'R,6R,6'R)-((16-amino-5,11,21,27-tetraoxo-14,18-dioxa-6,10,22,26-tetraazahentriacontane-1,31-diyl)bis(oxy))bis(5-acetamido-2-(acetoxymethyl)tetrahydro-2H-pyran-6,3,4-triyl)tetraacetic acid (21-8) as a colorless liquid. Yield: 0.850 g (crude); LCMS m / z 1206.20 [M+1] + .

[0591] To a stirred solution of (2R,2'R,3R,3'R,4R,4'R,5R,5'R,6R,6'R)-((16-amino-5,11,21,27-tetraoxo-14,18-dioxa-6,10,22,26-tetraazahentriacontane-1,31-diyl)bis(oxy))bis(5-acetamido-2-(acetoxymethyl)tetrahydro-2H-pyran-6,3,4-triyl)tetraacetic acid (21-8, 0.830 g, 0.688 mmol) in methanol (10.0 mL) was added sodium methanolate (25% methanol solution, 0.330 mL, 12.0 equivalents, 8.26 mmol) at room temperature, and the reaction mixture was stirred at the same temperature for 30 minutes. The reaction mixture was then quenched with 1N hydrochloric acid, and the solution was lyophilized to dryness to give N,N'-(10-amino-5,15-dioxo-8,12-dioxa-4,16-diazanonadecan-1,19-diyl)bis(5-(((2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)pentanamide) (21-9) as a light brown solid. Yield: 0.700 g, 98.10%. LCMS m / z 954.2 [M+1] + .

[0592] Dimethyl N,N'-(10-amino-5,15-dioxo-8,12-dioxa-4,16-diazanonadecan-1,19-diyl)bis(5-(((2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)pentanamide) (21-9, 0.200 g, 0.210 mmol) was added under stirring. To a solution of 2,3,4,5,6-pentafluorophenyl 12-oxo-12-(2,3,4,5,6-pentafluorophenyl)dodecanoate (9a, 0.172 g, 1.5 equivalents, 0.314 mmol) and N,N-diisopropylethylamine (0.0366 mL, 0.210 mmol) in methyl sulfoxide (2.00 mL) were added at room temperature, and the reaction mixture was stirred at the same temperature for 1 hour. The progress of the reaction was monitored by LC-MS, and after completion, the reaction mixture was purified by preparative HPLC (45-55% acetonitrile / water with 0.5% TFA buffer) to give 1-(((2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-16-((3-((3-(5-(((2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)pentanamido)propyl)amino)-3-oxopropoxy)methyl)-5,11,18-trioxo-14-oxa-6,10,17-triazanonacosan-29-oic acid perfluorophenyl ester (compound I-144) as a white solid. Yield: 0.026g, 9.0%; LCMS m / z 1332.98 [M+1] + ; 1H NMR (400 MHz, DMSO-d6 / D2O) δ 4.20 (d, J = 8.4 Hz, 2H), 3.92-3.90 (m, 2H), 3.67-3.62 (m, 6H), 3.55-3.50 (m, 3H), 3.41-3.38 (m, 3H), 3.35-3.25 (m, 8H), 3.05-2.95 (m, 8H), 2.74 (t, J = 7.2 Hz, 2H), 2.30-2.25 (m, 4H), 2.17 (t, J = 6.8 Hz, 1H), 2.05-1.98 (m, 6H), 1.85 (s, 6H), 1.68-1.60 (m, 2H), 1.55-1.38 (m, 16H), 1.37-1.14 (m, 15H).

[0593] Example 22: Synthesis of Compounds I-131 and I-133

[0594] N,N'-(10-(12-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)dodecanamido)-5,15-dioxo-8,12-dioxa-4,16-diazanonadecan-1,19-diyl)bis(5-(((2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)pentanamide) (Compound I-131) [ka]

[0595] Compound I-131 was synthesized by using the procedure described for compound I-144, substituting compound 27A for compound 9A, to give N,N'-(10-(12-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)dodecanamido)-5,15-dioxo-8,12-dioxa-4,16-diazanonadecan-1,19-diyl)bis(5-(((2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)pentanamide) (compound I-131) as a white solid. Yield: 0.026 g, 9.0%; LCMS m / z 1231.91 [M+1] + , 1 H NMR (400 MHz, DMSO-d6, D2O exchange) δ 6.94 (s, 1H), 4.20 (d, J = 8.4 Hz, 2H), 3.90 (t, J = 10.8 Hz, 1H), 3.70-3.63 (m, 4H ) 3.56-3.40 (m, 8H), 3.37-3.29 (m, 13H) 3.02-3.00 (m, 8H) 2.27 (t, J = 11.6 Hz, 4H), 2. 05 (t, J = 6.4 Hz, 4H), 1.79 (s, 6H), 1.49-1.41(m, 16H), 1.19 (s, 15H).

[0596] N-(2-(3-((3-(5-(((2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)pentanamido)propyl)amino)-3-oxopropoxy)ethyl)-12-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)dodecanamide (compound I-133) can be synthesized by adapting the procedure for compound I-131. The compound structure and data for compound I-133 are shown below. [ka]

[0597] LC-MS; m / z 770.43 [M+1] + .

[0598] 1 H NMR (400 MHz, DMSO-d6 (with D2O exchange)) δ 6.93 (s, 2H), 4.19 (d, J = 8.4 Hz, 1H), 3.60-3.57 (m, 2H), 3.57 (t, J = 6.4 Hz, 2H), 3.53-3.44 (m, 2H), 3.40-3.27 (m, 7H), 3.15-3.13 (m, 2H), 3.01 (brs, 4H), 2.28 (t, J= 6 Hz,2H), 2.02 (br t, J= 7 Hz, 4H), 1.78 (s, 3H), 1.46 (m, 9H), 1.18 (br m, 15H).

[0599] Example 23: Perfluorophenyl 1-(((2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-5,16-dioxo-9,12-dioxa-6,15-diazaheptacosane-27-oic acid (I-150) [ka]

[0600] A mixture of 5-[(2R,3R,4R,5R,6R)-3-acetamido-4,5-diacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl]oxypentanoic acid (Intermediate (Int)C) (1.00 equiv., 109 mg, 0.244 mmol) and HATU (1.45 equiv., 134 mg, 0.353 mmol) was dissolved in DMF (1.2 mL), followed by the addition of N,N-diisopropylethylamine (2.00 equiv., 85 μL, 0.487 mmol), followed by azido-PEG2-amine (1.10 equiv., 42 μL, 0.268 mmol). After 30 min, the reaction was filtered and then directly purified by reverse-phase HPLC (10-80% acetonitrile / water with 0.1% formic acid) to give [(2R,3R,4R,5R,6R)-5-acetamido-3,4-diacetoxy-6-[5-[2-[2-(2-azidoethoxy)ethoxy]ethylamino]-5-oxopentoxy]tetrahydropyran-2-yl]methyl acetate (Intermediate D). Yield: 136 mg, 92.5%. LCMS m / z 604.3 [M+H] + .

[0601] A solution of [(2R,3R,4R,5R,6R)-5-acetamido-3,4-diacetoxy-6-[5-[2-[2-(2-azidoethoxy)ethoxy]ethylamino]-5-oxo-pentoxy]tetrahydropyran-2-yl]methyl acetate (Intermediate D) (1.00 equiv., 130 mg, 0.215 mmol) in acetonitrile (1 mL) and THF (0.500 mL) was treated with triphenylphosphine (3.00 equiv., 169 mg, 0.646 mmol) for 2 hours, after which water (0.500 mL) was added. The reaction was stirred at room temperature overnight. The volatiles were removed under reduced pressure, and the residue was purified by reverse-phase HPLC (10-100% acetonitrile / water with 0.1% TFA) to give [(2R,3R,4R,5R,6R)-5-acetamido-3,4-diacetoxy-6-[5-[2-[2-(2-aminoethoxy)ethoxy]ethylamino]-5-oxopentoxy]tetrahydropyran-2-yl]methyl acetate (Intermediate E) as the TFA salt. Yield: 40 mg, 26%. LCMS m / z 578.2 [M+H] + .

[0602] A solution of [(2R,3R,4R,5R,6R)-5-acetamido-3,4-diacetoxy-6-[5-[2-[2-(2-aminoethoxy)ethoxy]ethylamino]-5-oxopentoxy]tetrahydropyran-2-yl]methyl 2,2,2-trifluoroacetate (Intermediate E) (1.00 equiv., 40.0 mg, 0.0463 mmol) in methanol (116 μL) was treated with sodium methoxide solution (0.75 M in methanol) (1.00 equiv., 62 μL, 0.0463 mmol) until a pH > 8 was reached (5 equiv.). Then, another portion of sodium methoxide solution (0.75 M in methanol) (1.00 equiv., 62 μL, 0.0463 mmol) was added, and the reaction was stirred at room temperature. After 24 hours, the reaction was acidified with a 4M solution of HCl in dioxane (1.00 equiv, 0.046 mL, 0.0463 mmol) and then the volatile components were removed under reduced pressure. The crude material was used in the next step without further purification. LCMS m / z 488.2 [M+H] + .

[0603] A slurry of 5-(((2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-N-(2-(2-(2-aminoethoxy)ethoxy)ethyl)pentanamide hydrochloride (Intermediate F) (1.00 equiv., 20.0 mg, 0.0410 mmol) and triethylamine (2.00 equiv., 11 μL, 0.0820 mmol) in DMF (0.439 mL) was added to a solution of bis(2,3,4,5,6-pentafluorophenyl)dodecanedioate (2.50 equiv., 57.6 mg, 0.102 mmol) in DMF (0.439 mL) and the reaction was stirred at room temperature. After 30 min, the reaction was diluted with DMSO, filtered, and then purified by reverse-phase HPLC (20-80% acetonitrile / water) to afford the title compound 12-[2-[2-[2-[5-[(2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]oxypentanoylamino]ethoxy]ethoxy]ethylamino]-12-oxododecanoic acid (2,3,4,5,6-pentafluorophenyl) (I-150) as a white solid. Yield: 13 mg, 34%. LCMS m / z 830.2 [M+H] + .

[0604] Compounds I-149 and I-151 can be obtained by adapting the synthetic procedure of compound I-150. The structures and LCMS data of I-149 and I-151 are shown below. [ka] LCMS m / z 786.2 [M+H] + . [ka] LCMS m / z 874.2 [M+H] + .

[0605] Example 24: Synthesis of perfluorophenyl 12-((3-(5-(((2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)pentanamido)propyl)amino)-12-oxododecanoate (I-164) [ka]

[0606] A solution of (2R,3R,4R,5R,6R)-5-acetamido-2-(acetoxymethyl)-6-((5-((3-aminopropyl)amino)-5-oxopentyl)oxy)tetrahydro-2H-pyran-3,4-diyl diacetate (ISP2-99-3a) (1.00 equiv., 80.0 mg, 0.159 mmol) in methanol (530 μL) was treated with a 1 M solution of sodium methoxide in methanol (1.40 equiv., 223 μL, 0.223 mmol). After 18 h, the reaction was acidified with 1 M aqueous HCl (0.500 equiv., 79 μL, 0.0794 mmol) until the solution was acidic (pH ∼3). The reaction was concentrated under reduced pressure to give crude 5-(((2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-N-(3-aminopropyl)pentanamide (Intermediate 3b) as the HCl salt. This material was used in the next step without further purification. Yield: 90 mg. LCMS m / z 378.2 [M+H] + .

[0607] A solution of 5-(((2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-N-(3-aminopropyl)pentanamide (Intermediate 3b) (1.00 equiv., 10.0 mg, 0.0265 mmol) and N,N-diisopropylethylamine (1.00 equiv., 4.6 μL, 0.0265 mmol) in DMF (265 μL) was added to an ice-cold solution of bis(2,3,4,5,6-pentafluorophenyl)dodecanedioate (2.00 equiv., 29.8 mg, 0.0530 mmol) in DMF (265 μL), and the reaction was stirred at 0° C. for 1 hour. The reaction solution was acidified with a drop of formic acid and then directly purified by reverse-phase HPLC (10-100% acetonitrile / water with 0.1% formic acid) to give perfluorophenyl 12-((3-(5-(((2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)pentanamido)propyl)amino)-12-oxododecanoate (I-164) as a white solid. Yield: 6.5 mg, 32%. LCMS m / z 778.34 [M+Na] + .

[0608] Example 25: 1-(4-(3-(((2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)propyl)-1H-1,2,3-triazol-1-yl)-13,13-bis(3-((2-(2-(2-(4-(3-(((2R,3R,4R,5R,6R)-3-acetamido Perfluorophenyl 4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)propyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl)amino)-3-oxopropyl)-10,15-dioxo-3,6-dioxa-9,14-diazahexacosane-26-oic acid (Compound No. I-135) [ka]

[0609] To a solution of perfluorophenyl 1-azido-13,13-bis(3-((2-(2-(2-azidoethoxy)ethoxy)ethyl)amino)-3-oxopropyl)-10,15-dioxo-3,6-dioxa-9,14-diazahexacosane-26-oic acid (131A, 1.0 equiv., 0.095 g, 0.086 mmol) in dimethyl sulfoxide (2.0 mL), N-((2R,3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)-2-(pent-4-yn-1-yloxy)tetrahydro-2H-pyran-3-yl)acetamide (XB7, 3.0 equiv., 0.074 g, 0.26 mmol) was added and stirred for 5 minutes. Tetrakis(acetonitrile)copper(I) hexafluorophosphate (8.4 equiv., 0.272 g, 0.729 mmol) was then added, and the reaction mixture was stirred at room temperature for 1 hour. After the reaction was complete, the reaction mixture was diluted with acetonitrile and purified by preparative HPLC (33-53% acetonitrile / water with 0.1% TFA). Fractions containing the desired product were combined and lyophilized to dryness to give 1-(4-(3-(((2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)propyl)-1H-1,2,3-triazol-1-yl)-13,13-bis(3-((2-(2-(2-(4-(3-(((2R,3R,4R,5R,6R)- 3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)propyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl)amino)-3-oxopropyl)-10,15-dioxo-3,6-dioxa-9,14-diazahexacosane-26-oic acid perfluorophenyl ester (compound number I-135) was obtained as an off-white solid. Yield: 0.036 g, 19.2%; LCMS m / z 978.89 [M+2H] ++ ; 1H NMR (400 MHz, DMSO-d6 / D2O) δ 7.76 (s, 3H), 4.42 (t, J = 5.2 Hz, 6H), 4.22 (d, J = 8.8 Hz, 3H), 3.75-3.68 (m, 11H), 3.63-3.62 (m, 3H), 3.54-3.46 (m, 13H), 3.43-3.42 (m, 8H), 3.40-3.37 (m, 4H), 3.35-3.24 (m, 10H), 3.12 (t, J = 5.6 Hz, 6H), 2.71 (t, J = 7.2 Hz, 2H), 2.61-2.57 (m, 6H), 2.05-1.92 (m, 7H), 1.79 (s, 9H), 1.76-1.73 (m, 10H), 1.62-1.60 (m, 2H), 1.45-1.41 (m, 2H), 1.36-1.29 (m, 2H), 1.25-1.16 (m, 10H).

[0610] Example 26: Perfluorophenyl 1-(4-((((2R,3R,4R,5R,6R)-5-acetamido-3,4-dihydroxy-6-methoxytetrahydro-2H-pyran-2-yl)methoxy)methyl)-1H-1,2,3-triazol-1-yl)-13,13-bis(3-((2-(2-(2-(4-((((2R,3R,4R,5R,6R)-5-acetamido-3,4-dihydroxy-6-methoxytetrahydro-2H-pyran-2-yl)methoxy)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl)amino)-3-oxopropyl)-10,15-dioxo-3,6-dioxa-9,14-diazahexacosane-26-oic acid (Compound No. I-136) [ka]

[0611] To a solution of 1-azido-13,13-bis(3-((2-(2-(2-azidoethoxy)ethoxy)ethyl)amino)-3-oxopropyl)-10,15-dioxo-3,6-dioxa-9,14-diazahexacosane-26-oic acid perfluorophenyl ester (132A, 1.0 equiv., 0.160 g, 0.146 mmol) in dimethyl sulfoxide (3 mL) was added N-((2R,3R,4R,5 (R,6R)-4,5-Dihydroxy-2-methoxy-6-((prop-2-yn-1-yloxy)methyl)tetrahydro-2H-pyran-3-yl)acetamide (XB3, 3.0 equiv., 0.120 g, 0.439 mmol) and tetrakis(acetonitrile)copper(I) hexafluorophosphate (8.4 equiv., 0.458 g, 1.23 mmol) were added, and the reaction mixture was stirred at room temperature for 1 h. After completion of the reaction, the reaction mixture was diluted with acetonitrile and purified by preparative HPLC (eluting from a C18 column with 30–57% acetonitrile / water with 0.1% TFA). Fractions containing the desired product were combined and lyophilized to dryness to give 1-(4-((((2R,3R,4R,5R,6R)-5-acetamido-3,4-dihydroxy-6-methoxytetrahydro-2H-pyran-2-yl)methoxy)methyl)-1H-1,2,3-triazol-1-yl)-13,13-bis(3-((2-(2-(2-(4-((((2R,3R,4R,5R,6R)-5- Acetamido-3,4-dihydroxy-6-methoxytetrahydro-2H-pyran-2-yl)methoxy)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl)amino)-3-oxopropyl)-10,15-dioxo-3,6-dioxa-9,14-diazahexacosane-26-oic acid perfluorophenyl ester (compound number I-136) was obtained as an off-white solid. Yield: 0.055 g, 19.6%; LCMS m / z 957.74 [M+2H] ++ ; 1H NMR (400 MHz, DMSO-d6) δ 8.04 (s, 3H), 7.81-7.80 (m, 2H), 7.63 (d, J = 8.8 Hz, 2H), 7.11 (s, 1H), 4.54 (d, J = 4.4 Hz, 5H), 4.50 (t, J = 5.2 Hz, 6H), 4.16 (d, J = 8.4 Hz, 3H), 3.80 (t, J = 5.2 Hz, 8H), 3.76-3.69 (m, 4H), 3.63-3.56 (m, 12H), 3.52-3.49 (m, 14H), 3.47-3.44 (m, 11H), 3.29 (s, 9H), 3.20 (s, 1H), 3.15 (d, J = 6.0 Hz, 8H), 2.76 (t, J = 6.8 Hz, 2H), 2.03-1.96 (m, 9H), 1.83-1.76 (m, 11H), 1.71-1.63 (m, 2H), 1.45-1.40 (m, 2H), 1.36-1.32 (m, 2H), 1.28-1.20 (m, 12H).

[0612] Example 27: 1-[4-(2-{[1,3-bis(2-{[3-(5-{[(2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}pentanamido)propyl]carbamoyl}ethoxy)-2-[(2-{[3-(5-{[(2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy} Pentanamido)propyl]carbamoyl}ethoxy)methyl]propan-2-yl]carbamoyl}ethyl)-1H-1,2,3-triazol-1-yl]-3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60,63,66,69,72-tetracosaoxapentaheptacontan-75-oic acid 2,3,4,5,6-pentafluorophenyl (Compound No. I-137) [ka]

[0613] To a solution of 1-azido-3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60,63,66,69,72-tetracosaoxapentaheptacontan-75-oic acid (27-1, 1.0 equiv., 0.050 g, 0.042 mmol) in dichloromethane (1.0 mL), pentafluorophenol (27-1a, 1.1 equiv., 0.008 g, 0.046 mmol) and N,N'-diisopropylcarbodiimide (1.5 equiv., 0.008 g, 0.064 mmol) were added, and the reaction mixture was stirred at room temperature for 2 h. After the reaction was complete, the reaction mixture was diluted with dichloromethane and filtered through a syringe filter. The filtrate was concentrated and dried to give 2,3,4,5,6-pentafluorophenyl 1-azido-3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60,63,66,69,72-tetracosaoxapentaheptacontan-75-oic acid (27-2) as a colorless sticky solid. Yield: 0.070 g (crude), LCMS m / z 669.8 [M+2H] ++ .

[0614] Acetic acid [(2R,3R,4R,5R,6R)-3,4-bis(acetyloxy)-6-{4-[(3-{3-[3-(2-{[3-(5)-{[(2R,3R,4R,5R,6R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}pentanamido)propyl]carbamoyl}ethoxy)-2-[(2-{[3-(5-{[(2R,3R,4R,5R,6R)-4,5-bis(acetyloxy)-6-[(acetyloxy)methyl]-3-acetamidooxan-2-yl]oxy}pentanamido)propyl]carbamoyl}ethoxy)methyl]-2-( To a solution of {pent-4-ynamido)propoxy]propanamido}propyl)carbamoyl]butoxy}-5-acetamidooxan-2-yl]methyl (27-2a, 1.0 equiv., 0.030 g, 0.016 mmol) in dimethyl sulfoxide (0.5 mL), 2,3,4,5,6-pentafluorophenyl 1-azido-3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60,63,66,69,72-tetracosaoxapentaheptacontan-75-oic acid 2,3,4,5,6-pentafluorophenyl (27-2, 2.0 equiv., 0.042 g, 0.032 mmol) was added and stirred for 5 min. Then, tetrakis(acetonitrile)copper(I) hexafluorophosphate (2.8 equiv., 0.016 g, 0.044 mmol) was added, and the reaction mixture was stirred at room temperature for 15 min. After the reaction was complete, the reaction mixture was diluted with acetonitrile and purified by preparative HPLC (27-62% acetonitrile / water with 0.1% TFA).Fractions containing the desired product were combined and lyophilized to dryness to give 1-[4-(2-{[1,3-bis(2-{[3-(5-{[(2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}pentanamido)propyl]carbamoyl}ethoxy)-2-[(2-{[3-(5-{[(2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl 2,3,4,5,6-Pentafluorophenyl tetracosaoxapentaheptacontan-75-oic acid (compound number I-137) was obtained as a colorless sticky solid. Yield: 0.012 g, 25.93%; LCMS m / z 1417.18 [M+2H]. ++ ; 1 H NMR (400 MHz, D2O) δ 7.82 (s, 1H), 4.56 (bs, 3H), 4.40 (d, J = 8.4 Hz, 4H), 3.90-3.82 (m, 14H), 3.75-3.70 (m, 5H), 3.67-3.49 (m, 111H), 3.17 (d, J = 6.4 Hz, 12H), 3.05 (t, J = 5.2 Hz, 2H), 2.92 (t, J = 7.6 Hz, 2H), 2.57 (t, J = 6.0 Hz, 2H), 2.43 (bs, 6H), 2.20 (bs, 6H), 1.99 (s, 9H), 1.71-1.66 (m, 6H), 1.54 (bs, 12H).

[0615] Example 28: 11-{[1,5-bis({[2-(2-{2-[4-(3-{[(...

Claims

1. Formula (I): 【Chemical 238】 (wherein,[ Y is the target moiety, n is from 1 to 500, L is a linker, X is of formula (Ia): 【Chemical 239】 (wherein,[ R １ is selected from -OH, -OC(O)R, -C(O)NHR, -Z １ -*, and optionally substituted triazole, provided that R is optionally substituted C １～６ alkyl or optionally substituted aryl, and R ２ is selected from -NHCOCH ３ , -NHCOCF ３ , -NHCOCH ２ CF ３ , -OH, optionally substituted triazole, and -Z １ -*, and R ３ is selected from -H, -OH, -CH ３ , -OCH ３ , -OCH ２ CH=CH, and -Z １ -*, and is selected from R １ ~R ３ One of them is -Z １ -*, or contains -Z １ -*, where "*" represents the bonding point of Z to the linker (L) １ and R ４ and R ５ are each independently selected from H and a promoiety, or R ４ and R ５ are cyclically linked to form a promoiety, R １１ is a group that forms a crosslink to H or the carbon atom in the 1-position, Z １ is Z １１ , optionally substituted Z １１ -heteroaryl, optionally substituted Z １１ -aryl, optionally substituted heteroaryl, optionally substituted aryl, optionally substituted alkyl, optionally substituted amide, optionally substituted sulfonamide, optionally substituted urea, and optionally substituted thiourea, and is a linking moiety selected from Z １１ is selected from -O-, -S-, NR ２１ -, and -C(R ２２ ) ２ and is selected from Each R ２１ is independently selected from H and optionally substituted (C １ ~C ６ ) alkyl Each R ２２ is independently selected from H, halogen (e.g., F), and optionally substituted (C １ -C ６ ) alkyl) is the moiety that binds to the cell surface asialoglycoprotein receptor (ASGPR), i) n is 3, and R １ is OH, and R ２ is -NHCOCH ３ and R ４ and R ５ are H, and when R ３ is Z １ then Z １ is not O, ii) n is 2 or 3, and R １ is OAc, and R ２ is -NHCOCH ３ and R ４ and R ５ are Ac, and R ３ is Z １ when it is the case, Z １ is not O, iii) n is 2 or 3, and R １ is Obz, and R ２ is -NHCOCH ３ and R ４ and R ５ are Bz, and R ３ is Z １ when it is the case, Z １ is not O, iv) n is 3, R １ is OH, R ２ is -NHCOCH ３ and R ４ and R ５ are H, R ３ is Z １ and when Z １１ is O, L contains a main chain of at least 16 consecutive atoms up to the branch point, v) n is 3, and R １ is Z １ wherein, Z １ is O, and when R ４ and R ５ are H, R ３ is -CH ３ -OCH ３ or -OCH ２ CH=CH, instead vi) R １１ is a group of the formula -CH ２ O- that forms a crosslink to the second carbon atom, and R ２ is -NHCOCH ３ and when R ４ and R ５ are H, then R １ and R ３ are not Z １ (not in Z) a cell surface ASGPR-binding compound, or a prodrug thereof, or a salt thereof.

2. Each X is independently of the formula: 【Chemical 240】 one of, the compound according to claim 1, or a prodrug thereof, or a salt thereof.

3. Z １ is Z １１ -Ar, provided that Ar is heteroaryl optionally substituted or aryl optionally substituted, Optionally Z11 is O, S, or C(R22)2, Ar is a monocyclic 5- or 6-membered heteroaryl or aryl, Z1 is preferably -C(R22)2-triazole-, more preferably 【Chemical 241】 or Optionally Z1 is a monocyclic 5- or 6-membered heteroaryl or aryl, preferably Z1 is 【Chemical 242】 or Optionally Z1 is -O-, -S-, -C(R22)2-, -NR21-, -CONR21-, and 【Chemical Formula 243】 (wherein,[ X1 is O or S, t is 0 or 1, R21 and each R23 are independently selected from H and optionally substituted (C1-C6)alkyl, each R22 is independently selected from H, halogen, and optionally substituted (C1-C6)alkyl) selected from, preferably Z1 is -S-, -O- or -CH2-, the compound according to claim 2, or a prodrug thereof, or a salt thereof.

4. n is 1, and L contains a straight-chain linker having a backbone of 20 or more consecutive atoms that covalently link X to Y via Z １ and L preferably contains a backbone of 20 to 100 consecutive atoms, preferably 25 or more consecutive atoms, preferably 30 or more consecutive atoms, or n is 2 or more, preferably 2 or 3, and L is a branched-chain linker that covalently links two or more X moieties to Y via a linking moiety Z1, each branched chain of L covalently links each X moiety to the branch point of the linker L via Z1, optionally containing a straight-chain linker of 14 or more contiguous atoms, each branched chain of L preferably containing a straight-chain linker of 14 to 50 contiguous atoms, preferably 20 or more contiguous atoms, the compound according to claim 2, or a prodrug thereof, or a salt thereof.

5. L is of formula (II): 【Chemical 244】 L as in, wherein,[ L １ and L ３ are each independently a linker, and L ２ is a branched-chain linking moiety, and L １ to L ３ collectively provide a straight-chain or branched-chain linker between X and Y a, b, and c are independently 0 or 1, ** is Z １ The junction of X to L １ via is represented by, *** represents the point of attachment to Y, when n is 1, a is 1 and b is 0, when n > 1, a is 1 and b is 1, Optionally, each of L1 to L3 is independently selected from -C1-20-alkylene-, -NHCO-C1-6-alkylene-, -CONH-C1-6-alkylene-, -NHC1-6-alkylene-, -NHCONH-C1-6-alkylene-, -NHCSNH-C1-6-alkylene-, -C1-6-alkylene-NHCO-, -C1-6-alkylene-CONH-, -C1-6-alkylene-NH-, -C1-6-alkylene-NHCONH-, -C1-6-alkylene-NHCSNH-, -O(CH2)p-, -(OCH2CH2)p-, -NHCO-, -CONH-, -NHSO2-, -SO2NH-, -CO-, -SO2-, -O-, -S-, pyrrolidine-2,5-dione, 1,2,3-triazole, -NH-, and -NMe- (where each p is independently 1 to 50), and includes one or more linking moieties independently selected therefrom, Optionally, L is a repetition of ethylene glycol moieties, preferably including 1 to 25 ethylene glycol moieties, Optionally, L is one or more 1,2,3-triazole linking moieties, and optionally has the following structure: 【Chemical 245】 (wherein w1, u1, and q1 are independently 1 to 25) and includes one or more 1,2,3-triazole moieties selected therefrom, When n is 2 or more, L2 is optionally 【Chemical 246】 (wherein each x and y is independently 1 to 10) selected from, Optionally, L1 to L2 include a main chain of 14 or more consecutive atoms between X and the branching atom, Optionally, L3 includes a main chain of 10 to 80 consecutive atoms, preferably including a linking moiety selected from (C10-C20)-alkylene or -(OCH2CH2)p- (where p is 1 to 25), Optionally, the linker of the formula (II) includes 20 to 100 consecutive atoms, preferably 25 or more consecutive atoms, preferably 30 or more consecutive atoms, Optionally, -Z1-L1- is 【Chemical 247】 (wherein o, p, q, r, s, t, u, v, w, x, y, z, and z1 are each independently 1 to 6) selected from the groups, preferably -Z1-L- includes 【Chemical 248】 a group selected from the groups, the compound according to claim 1, or its prodrug, or its salt.

6. At least one X is of formula (Ic): 【Chemical 249】 and is Optionally, Z1 is selected from -O-, -S-, -CONR21-, and optionally substituted -(C(R22)2)q-heteroaryl (where q is 0 or 1), preferably Z1 is -O- or optionally substituted -(C(R22)2)q-triazole (where q is 0 or 1), preferably Z1 is 【Chemical 250】 and Optionally, n is 1, and L contains a linear linker having a main chain of 20 or more consecutive atoms that covalently links X to Y via Z1, Optionally, n is 2 or more, and L is a branched-chain linker that covalently links two or more X moieties to Y, and each branched chain of L covalently links each X moiety to the branch point of the linker L via Z1 and contains a linear linker of 14 or more consecutive atoms. The compound according to claim 1, or a prodrug thereof, or a salt thereof.

7. At least one X is of formula (Id): 【Chemical 251】 and Optionally, Z1 is selected from optionally substituted -(C(R22)2)q-heteroaryl (where q is 0 or 1) and 【Chemical 252】 preferably Z1 is optionally substituted -(C(R22)2)q-triazole (wherein q is 0 or 1), preferably Z1 is 【Chemical 253】 and Optionally, Z1 is 【Chemical 254】 (wherein R23 is H or C(1-3)-alkyl) and Optionally, R3 is H; or At least one X is of formula (Ie): 【Chemical 255】 wherein Z2 is absent or selected from -O-, -S-, NR25-, and -C(R22)2, Ring A is absent or selected from 5- or 6-membered optionally substituted aryl and 5- or 6-membered optionally substituted heteroaryl, Z3 is a linking moiety selected from Z12, optionally substituted alkyl, optionally substituted Z12-alkyl, optionally substituted Z12-heteroaryl, optionally substituted Z12-aryl, optionally substituted heteroaryl, optionally substituted aryl, optionally substituted amide, optionally substituted sulfonamide, optionally substituted urea, and optionally substituted thiourea. Z12 is selected from -CH2O-, -O-, -S-, -NR26-, and -C(R22)2-, R25 and R26 are each independently selected from H, optionally substituted (C1-C6) alkyl (e.g., C(1-3)-alkyl such as methyl), and optionally substituted acyl, each R22 is independently selected from H, halogen (e.g., F), and optionally substituted (C1-C6) alkyl, optionally formula (Ie) is one of formulae (If)-(Ih): 【Chemical 256】 being any one of them, optionally said ring A is a 5- or 6-membered heteroaryl, preferably selected from triazole, pyridine, pyrimidine, pyridazine, pyrazine, triazine, thiadiazole, thiophene, oxazole, isoxazole, isothiazole, thiazole, oxadiazole, and furan, preferably selected from triazole, pyrimidine, and thiadiazole, optionally formula (Ie) is one of formulae (Ij)-(Im): 【Chemical 257】 being any one of them, wherein, Y1 to Y3 are each independently N or CR27, R24 and R27 are each independently selected from H, optionally substituted C(1-6)-alkyl, optionally substituted fluoroalkyl, and halogen, Z3 is preferably selected from -O-, -CH2O-, -OCH2-, optionally substituted -OCH2-heteroaryl, optionally substituted -OCH2-aryl, optionally substituted -CH2O-heteroaryl, and optionally substituted -CH2O-aryl, more preferably 【Chemical 258】 selected from, optionally Z1 is -NR23CO- (wherein R23 is H or C(1-3)-alkyl), optionally n is 1, and L contains a linear linker having a main chain of 20 or more consecutive atoms that covalently links X to Y via Z1, Optionally, n is 2 or more, and L is a branched-chain linker that covalently links two or more X moieties to Y, and each branched chain of L covalently links each X moiety to the branch point of the linker L via Z1, The compound according to claim 1, or a prodrug thereof, or a salt thereof, comprising a linear linker of 14 or more consecutive atoms.

8. R １ is OH, or R1 is -OC(O)R or 【Chemical 259】 The compound according to claim 1, or a prodrug thereof, or a salt thereof.

9. R ２ is -NHCOCH ３ or R2 is -NHCOCF3, or R2 is -NHCOCH2CF3, or R2 is -OH, or R2 is 【Chemical 260】 The compound according to claim 1, or a prodrug thereof, or a salt thereof.

10. R ４ and R ５ at least one of which is a pro component, the pro component being optionally an ester, the ester being optionally of the formula -OCOCH3, -OCOCH(CH3)2, or -OCOC(CH3)3, or R4 and R5 being cyclically linked to form a pro component, optionally of the formula (Io) or (Ip): 【Chemical 261】 (wherein Y4 is a counter ion) forms a pro component, or The compound according to claim 1, or a prodrug thereof, or a salt thereof, wherein both R4 and R5 are H.

11. Each X has the following structure: 【Chemical 262】 selected from one of, wherein R ６ and R ２３ are independently H or (C １～３ ) alkyl, optionally n is 1, and X is 【Chemical 263】 or Each X has the following structure: 【Chemical 264】 is selected from one of, wherein R5 and R4 are independently H or a pro component, or R5 and R4 are cyclically linked to form a pro component, n1 and n2 are each independently an integer from 1 to 6, Y4 is a counter ion, or Each X has the following structure: 【Chemical 265】 The compound according to claim 1, or a prodrug thereof, or a salt thereof, which is selected from one of.

12. Y is selected from a small molecule, a dye, a fluorophore, a monosaccharide, a polysaccharide, a lipid, a protein, a polynucleotide, an enzyme, an enzyme substrate, a polymer, and a chemoselective linking group, or a precursor thereof, or Y is a moiety that specifically binds to an extracellular target protein, and the target protein is optionally a membrane-bound protein or a soluble extracellular protein, Optionally, Y is a target-binding small molecule, Optionally, Y is a target-binding biomolecule, and the biomolecule is optionally selected from a peptide, a protein, a glycoprotein, a polynucleotide, an aptamer, and an antibody or antibody fragment, or Y is optionally an antibody, an antibody fragment, a chimeric fusion protein, a modified protein domain, and a D-protein binder of a target protein, Optionally, Y is an antibody or antibody fragment that specifically binds to a target protein, and the compound is a conjugate of formula (III): 【Chemical 266】 wherein, n is from 1 to 20, optionally from 1 to 6, optionally at least 2, optionally 2, optionally 3, and m is from 1 to 80, optionally from 1 to 20, optionally 10 or less, optionally from 2 to 8, optionally from 2 to 6, optionally an average load number of about 4, and each X is a moiety that binds to the cell surface ASGPR, each L is a linker, each Z is a residual moiety derived from the covalent linking of a chemoselective linking group to a compatible group of the Ab, optionally a residual moiety derived from the covalent linking of a thiol-reactive chemoselective linking group to one or more cysteine residues of the Ab, or a residual moiety derived from the covalent linking of an amine-reactive chemoselective linking group to one or more lysine residues of the Ab, Ab is an antibody or antibody fragment that specifically binds to the target protein, optionally the antibody or antibody fragment is an IgG antibody and / or a humanized antibody, and optionally the antibody or antibody fragment specifically binds to a secreted or soluble target protein or specifically binds to a cell surface receptor target, optionally each X is independently of the formula (Ib): 【Chemical 267】 or each X is optionally 【Chemical 268】 selected from or each X is optionally 【Chemical 269】 wherein R5 and R4 are independently H or a pro component, or R5 and R4 are cyclically linked to form a pro component), or each X is independently of the formula (Ie): 【Chemical 270】 or n is 1 and X is 【Chemical 271】 The compound according to claim 1, or a prodrug thereof, or a salt thereof.

13. A composition for use in a method of internalizing a target protein into a cell comprising a cell surface asialoglycoprotein receptor (ASGPR), said composition comprising a compound according to any one of claims 1 to 12, or a prodrug thereof, or a salt thereof, said method comprising contacting a cell sample comprising said cell and said target protein with said compound, said prodrug, or said salt, said compound, said prodrug, or said salt specifically binding to said target protein and specifically binding to said cell surface receptor to promote cellular uptake of said target protein, optionally said target protein is a membrane-bound protein or an extracellular protein; The composition, optionally, wherein the compound comprises an antibody or antibody fragment (Ab) that specifically binds to the target protein.

14. A composition for use in a method of reducing the level of a target protein in a biological system, the composition comprising a compound according to any one of claims 1 to 12, or a prodrug thereof, or a salt thereof, the method comprising contacting the biological system with the compound, the prodrug, or the salt, wherein the compound, the prodrug, or the salt specifically binds to the target protein and specifically binds to a cell surface receptor of a cell in the biological system to promote cellular uptake and degradation of the target protein, the biological system optionally comprising cells comprising a cell surface asialoglycoprotein receptor (ASGPR), the biological system optionally being a human subject, the biological system optionally being an in vitro cell sample, the target protein optionally being a membrane-bound protein, the target protein optionally being an extracellular protein, the composition.

15. A composition for treating a disease or disorder associated with a target protein in a subject in need thereof, the composition comprising a compound according to any one of claims 1 to 12, or a prodrug thereof, or a salt thereof, wherein the compound, the prodrug, or the salt specifically binds to the target protein, the disease or disorder optionally being selected from the group consisting of an inflammatory disease, an autoimmune disease, or cancer, the composition.

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