Cycling Lysosome-Targeted Antibody Conjugates

Abstract

The present disclosure provides conjugates comprising a moiety that targets a lysosomal targeting molecule, a linker, and an antibody that specifically binds to a cell surface or extracellular target molecule targeted for degradation. The present disclosure provides a class of conjugate compounds comprising a ligand moiety that binds to a lysosomal targeting molecule (e.g., a receptor) and an antibody or antibody fragment that binds to a cell surface containing a transmembrane or extracellular target molecule. The conjugates promote degradation of the target and can be repeatedly "cycled" into and out of the cell.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. 63 / 480,510, filed January 18, 2023, under 35 U.S.C. §119(e), the entire contents of which are incorporated herein by reference. [Background technology]

[0002] Many therapeutic drugs act by binding to functionally important sites in target proteins to modulate their activity or by recruiting immune effectors to act on target proteins, as with many monoclonal antibody drugs. However, there is an untapped reservoir of medically important human proteins that are considered "undruggable" because they do not easily fit into currently available therapeutic targeting approaches. Therefore, there is a need for therapeutics that can target a broader range of proteins. Summary of the Invention [Means for solving the problem]

[0003] The present disclosure provides a class of conjugate compounds comprising a ligand moiety that binds to a lysosomal targeting molecule (e.g., a receptor) and an antibody or antibody fragment that binds to a cell surface, including a transmembrane or extracellular target molecule. The conjugates can be repeatedly "cycled" into and out of cells, promoting target degradation. Such cycling allows for sustained activity on the order of hours to days, allowing a single conjugate to promote lysosomal degradation of multiple targets. In some embodiments, binding of the ligand moiety to the lysosomal targeting molecule can cause internalization of the lysosomal targeting molecule and the conjugate. In one embodiment, a ligand moiety conjugated to a target binding site via a linker is provided, wherein the ligand moiety binds to the lysosomal targeting molecule extracellularly, the target binding moiety binds to the target molecule extracellularly, the target binding moiety dissociates from the target molecule in an endosome, and the conjugate is externalized from the cell. In some embodiments, the conjugate comprises a target-binding moiety that has a higher binding affinity for the target molecule at extracellular pH than at endosomal pH, and simultaneously has one or more of: a) a target-binding moiety that has a higher affinity for FcRn at endosomal pH than at extracellular pH; b) a ligand moiety X that has equal binding affinity for the lysosomal-targeted molecule extracellularly and in the endosome; c) a ligand moiety X that has equal binding affinity for the lysosomal-targeted molecule at extracellular and endosomal pH; and / or d) a ligand moiety X that has equal binding affinity for the lysosomal-targeted molecule at extracellular and endosomal Ca2+ concentrations. In some embodiments, the conjugate exhibits a duration of activity on the order of hours to days.

[0004] In some embodiments, the conjugates are configured to exhibit enhanced activity and / or superstoichiometric clearance of the target in a biological system. The conjugates described herein can facilitate transport of the target molecule into a cell and can also facilitate sequestration and / or degradation of the target molecule of interest in the lysosomes of the cell.

[0005] Also provided herein are compositions comprising such conjugates and methods of using the conjugates to sequester and / or target a polypeptide or molecule of interest for lysosomal degradation, as well as methods of using the conjugates.

[0006] These and other features, aspects, and advantages of the present invention will become better understood with reference to the following description and accompanying drawings. [Brief explanation of the drawings]

[0007] [Figure 1] (FIG. 1A) Schematic of the mechanism hypothesized to occur after administration of the conjugates described herein, where externalization of the conjugates from cells is mediated by binding to FcRn. (FIG. 1B) Schematic of the mechanism hypothesized to occur after administration of the conjugates described herein, where externalization of the conjugates from cells is mediated by persistent binding to a lysosomal targeting molecule. [Figure 2] 1 shows a graph of serum human IgE (hIgE) concentration versus time following administration of Oma or the Oma conjugate Oma-1226 to mice that had been treated with hIgE 24 hours previously. [Figure 3] 1 shows a graph of hIgE concentration in mouse serum versus time after hIgE administration in mice treated with Oma or Oma-1226 24 hours previously. [Figure 4] 1 shows a graph of hIgE concentration in mouse serum versus time following administration of hIgE to untreated mice or mice treated 24 hours prior with Oma or Oma-1228. [Figure 5] 1 shows a graph of hIgE concentration in mouse serum versus time following administration of a superstoichiometric amount of hIgE to untreated mice or to mice treated 24 hours previously with Oma or Oma-L443C-1117. [Figure 6]Graph of hIgE concentration in mouse serum versus time after administration of a superstoichiometric amount of hIgE to untreated mice or to mice treated 24 hours prior with Oma or Oma-1119 (DAR4 lysine conjugates). [Figure 7] 1 shows a graph of hIgE concentration in mouse serum versus time after administration of two doses of hIgE to untreated mice or mice previously treated with Oma-L443C or Oma-L443C-1117. [Figure 8] A–B show graphs of serum hIgE concentrations versus time after administration of IgE to FcRn wild-type mice ( Fig. 8A ) or FcRn knockout mice ( Fig. 8B ) that were untreated or previously treated with Oma-L443C or Oma-L443C-1120. [Figure 9] 1 shows the cycling of various ASGPR-based conjugates. [Figure 10] 1 shows the cycling of various M6PR-based conjugates. [Figure 11] 1 shows the mechanism of FcRn-mediated cycling. [Figure 12] 1 shows a graph of the percentage ratio of hIgE concentration in the serum of myeloma-implanted mice to the initial hIgE concentration versus time after administration of Oma S35H / Y57H-2328 and Oma S35H / Y57H-2327. [Figure 13] 1 shows a graph of hC5 concentration in mouse serum versus time after administration of hC5 followed by administration of Ecu-2405, Ecu-1117, ALXN1210-2405, and ALXN1210-1117. DETAILED DESCRIPTION OF THE INVENTION

[0008] Target-binding conjugates The present disclosure provides a class of conjugates comprising a ligand moiety X that binds to a lysosomal targeting molecule conjugated to a target-binding moiety Y, and an antibody that specifically binds to a cell surface or extracellular target molecule. In some embodiments, binding of the ligand moiety to the lysosomal targeting molecule can cause internalization of the lysosomal targeting molecule and the conjugate. In some embodiments, the antibody conjugate is configured to exhibit enhanced activity and / or superstoichiometric clearance of the target in a biological system. "Superstoichiometric clearance" refers to a conjugate in which a greater than stoichiometric ratio of target molecule is removed from the extracellular environment relative to the amount of conjugate administered, i.e., the conjugate promotes degradation of a molar excess of the target relative to the conjugate. The compound or conjugate binds to the target molecule extracellularly, internalizes the target molecule, and releases the target molecule, resulting in destruction of the target molecule in the lysosome and release of the compound or conjugate to the outside, allowing the process to be repeated. Thus, the proportion of compound or conjugate administered will be less than the amount of target molecule cleared or removed from the extracellular environment.

[0009] In one embodiment, a ligand moiety X conjugated to a target-binding moiety Y via a linker L is provided, wherein the ligand moiety X binds to a lysosomal targeting molecule extracellularly, the target-binding moiety Y binds to the target molecule extracellularly, the target-binding moiety Y dissociates from the target molecule in an endosome, and the conjugate is externalized from the cell. In some embodiments, Y is an antibody or antibody fragment.

[0010] In some embodiments, the target-binding conjugate has formula (I'): [ka] or a prodrug thereof, or a pharmaceutically acceptable salt thereof, wherein: X is a ligand moiety that binds to a lysosomal targeting molecule; n is 1 to 20; L is a linker, m is 1 to 10; Y is an antibody or antibody fragment that specifically binds to a cell surface or extracellular target molecule; The conjugate is configured to extend the duration of the conjugate in its active form.

[0011] In some embodiments, the conjugate is configured to promote degradation of the target molecule for at least 1 day, or at least 2 days, or at least 4 days, or at least 7 days after administration of the conjugate.

[0012] Previously reported lysosome-targeting chimeras are based on O-linked GalNAc structures (see, e.g., G. Ahn et al., Nat. Chem. Biol. 2021, 17(9)937-46), and while they are capable of promoting lysosomal degradation of target molecules, they are chemically and enzymatically unstable in the endosomal environment and are rapidly degraded. In contrast, the conjugates described herein are stable in the endosomal environment, as evidenced, for example, by their ability to promote target degradation for more than 24 hours. Compare the results shown in Figures 3 and 4, described below.

[0013] The conjugates described herein are believed to exhibit enhanced activity or superstoichiometric clearance of targets in biological systems, or both. In some embodiments, the loading and / or stability of antibody conjugates can be provided by the use of conjugation chemistries to specific sites on the antibody, e.g., cysteine-reactive chemoselective conjugation chemistries.

[0014] In some embodiments, the ligand is M6PR or ASGPR.

[0015] In some embodiments of Formula (I'), n is 1 to 10, e.g., 1 to 6, 1 to 4, or 1 to 3. In some embodiments, X is an M6PR ligand moiety and n is 1 to 6, e.g., n is about 4 (i.e., the average loading is about 4).

[0016] In some embodiments, X is an ASGPR ligand moiety and n is 1 to 3, for example, 1 to 2. In some embodiments, n is 1. In some embodiments, n is 2.

[0017] In some embodiments of Formula (I'), m is 1 to 6, or 1 to 4. It is understood that depending on the conjugation chemistry, m can refer to a discrete number, or m can refer to an average, for example, the loading of ligand linkers on an antibody (i.e., the DAR ratio).

[0018] In some embodiments, Y is selected from a human antibody, a humanized antibody, or a chimeric antibody. The Y antibody can have an Fc region or Fc domain that enables binding to FcRn (neonatal fragment crystallizable receptor) in a target cell of interest to promote internalization and / or degradation activity of the conjugate within the cell. In some embodiments, the Fc region of an antibody in a conjugate of the present disclosure comprises one or more mutations that improve binding affinity to FcRn compared to a reference Fc region lacking the one or more mutations.

[0019] In some embodiments, antibody Y may bind to calcium (Ca), e.g., to promote release of the conjugate from an intracellular molecule. 2+ In some embodiments, antibody Y is selected to have a binding affinity for the cell surface or extracellular target molecule that is pH- or pH-dependent. In some embodiments, antibody Y is selected to have a higher binding affinity for the cell surface or extracellular target molecule at neutral pH than at low pH.

[0020] In some embodiments of Formula (I), the conjugate has Formula (II): [ka] wherein: L 1 and L 3 are independently a linker (e.g., as described herein), and L 2 is a branched linking moiety (e.g., as described herein), and L 1 ~L 3 together provide a linear or branched linker between X and Y, a, b, and c are independently 0 or 1; Z is a residue moiety resulting from the covalent bond between a chemoselective ligation group and a compatible group of Y (e.g., as described herein), wherein: If n is 1, a is 1 and b is 0, If n>1, a is 1 and b is 1.

[0021] In some embodiments, the target-binding conjugate has the formula (IIa'): [ka] or a prodrug thereof, or a salt thereof, During the ceremony, n is 1 to 3, m is 1 to 3; X and Y are each independently as defined herein; L 1 ~L 6 are each independently a linking moiety, and together they form Z 1 and Y, forming a linear or branched linker therebetween; a, b, c, d, and e are each independently 1, 2, 3, 4, or 5.

[0022] Sosomal targeting molecules and ligand moieties Lysosomal targeting molecules are cell surface receptors that allow for internalization of the conjugated compounds of the present disclosure.

[0023] The term "ligand moiety" refers to the portion of a conjugate described herein that binds to a lysosomal targeting molecule.

[0024] In some embodiments, the lysosomal targeting molecule is selected from asialoglycoprotein receptor (ASGPR), cation-independent mannose-6-phosphate receptor (CI-M6PR, also referred to herein as M6PR), folate receptor, CD63, sortilin, IFITM3, a molecule in the endosomal / lysosomal pathway, LIMP-1, and LIMP-2. In some embodiments, the lysosomal targeting molecule is ASGPR. In some embodiments, the lysosomal targeting molecule is CI-M6PR. In some embodiments, the lysosomal targeting molecule is a folate receptor.

[0025] A variety of ligand moieties (compounds or moieties that bind to lysosomal targeting molecules) can be utilized in the conjugate compounds of the present disclosure.

[0026] In some embodiments, the ligand moiety comprises a monosaccharide. In some embodiments, the ligand moiety comprises galactose. In some embodiments, the ligand moiety comprises mannose. In some embodiments, the ligand moiety comprises pyranose. In some embodiments, the ligand moiety is not an antibody or antibody fragment.

[0027] Ligand moieties that bind to ASGPR are described in International Publication WO2023 / 288033, filed July 14, 2022, the entire disclosure of which is incorporated herein by reference.

[0028] Ligand moieties that bind to CI-M6PR are described in International Publication WO2023 / 288015, filed July 14, 2022, the entire disclosure of which is incorporated herein by reference.

[0029] Ligand moieties that bind to folate receptors are described in International Publication WO2022 / 150721, filed January 10, 2022, the entire disclosure of which is incorporated herein by reference.

[0030] In some embodiments of any of the conjugates disclosed herein, the targeting moiety is located at the C1 carbon atom of X (i.e., the R 1 When X is bonded to the linker (L) at the anomeric carbon (e.g., via an -O-), the atom directly bonded to the C1 carbon atom of X is not -O-.

[0031] ASGPR ligand moiety The present disclosure provides a class of compounds that comprise a ligand moiety that specifically binds to ASGPR.The ASGPR ligand moiety of the present disclosure can be linked to various target moieties without affecting the specific binding and function of cell surface ASGPR.The present inventors have demonstrated that the compounds of the present disclosure can utilize the function of cell surface ASGPR in biological systems, for example, by internalizing and sequestrating compounds into lysosomes of cells, and in some cases subsequently degrading them in lysosomes.The compounds of the present disclosure can be used in various applications.

[0032] The term "asialoglycoprotein receptor" (ASGPR), also known as the 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).

[0033] A compound or moiety or conjugate comprising such an ASGPR-binding moiety (X) (e.g., as described herein) may bind to other receptors, or may bind with lower affinity, for example, as determined by immunoassay or other assays known in the art. In one embodiment, X, or a compound, moiety, or conjugate described herein comprising such X, specifically binds to a cell surface ASGPR with an affinity of 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 one embodiment, X, or a compound or moiety described herein comprising X, specifically binds to a cell surface ASGPR with an affinity (K) of 20 mM or less. d ) specifically binds to ASGPR. In some embodiments, such binding is 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. The terms "bind," "bind," "specifically bind," or "specifically bind to" are used interchangeably.

[0034] The ASGPR-binding compounds or ASGPR ligand moieties of the present disclosure comprise a moiety (X) that specifically binds to the cell surface receptor ASGPR. The ASGPR-binding compounds or ASGPR ligand moieties may be monovalent or multivalent (e.g., bivalent or trivalent or higher valency), with monovalent compounds comprising a single ASGPR ligand moiety and multivalent compounds comprising two or more such moieties.

[0035] In certain embodiments, the ASGPR-binding moiety or ASGPR-ligand moiety X can bind to a specific cell-surface ASGPR and direct (or target) the molecule to this receptor. In certain embodiments, the ASGPR-binding moiety or ASGPR-ligand moiety X can bind to an ASGPR and direct (or target) a compound or conjugate described herein to lysosomes for internalization and sequestration and / or subsequent lysosomal degradation.

[0036] In some embodiments, the ASGPR binding moiety or ASGPR ligand moiety, X, comprises an amino sugar ring derivative of galactose (e.g., N-acetylgalactosamine and its analogs) linked via a linking moiety to the 1-, 6-, or 2-position of the sugar ring. The linking moiety can be 1 to 10 atoms, e.g., 1 to 6, or 1 to 5, 1 to 4, or 1 to 3 atoms in length. In some embodiments, the amino sugar ring derivative of galactose is linked via a linking moiety to the oxygen, sulfur, nitrogen, or carbon atom at the 1-position of the ring. In some embodiments, the amino sugar ring derivative of galactose is linked via a linking moiety to the oxygen, sulfur, nitrogen, or carbon atom at the 6-position of the ring. In some embodiments, the amino sugar ring derivative of galactose is linked via a linking moiety to the oxygen, sulfur, nitrogen, or carbon atom at the 2-position of the ring. In certain embodiments, the amino sugar derivative of galactose is linked via a linking moiety to a heteroaryl group at the 1-, 6-, or 2-position of the ring. In certain embodiments, the amino sugar derivative of galactose is a bicyclic structure.

[0037] In some embodiments, the ASGPR binding compound or ASGPR ligand moiety is monovalent (e.g., in Formula (I), n is 1), such that the ASGPR binding compound or ASGPR ligand moiety comprises a single ASGPR ligand moiety (X) linked to a moiety of interest or target binding moiety (Y) via a linking moiety L at position 1, 6, or 2 of (X). In certain embodiments of Formula (I), n is 1 and L comprises a linear linker having a backbone of 20 or more consecutive atoms covalently linking the ASGPR ligand (X) to Y via a linking moiety at either position 1, 2, or 6 of X. In some instances, n is 20 to 100, e.g., 25 to 80, 25 to 60, or 25 to 50 consecutive atoms. In certain embodiments of Formula (I), n is 1 and L comprises a backbone of 25 or more consecutive atoms covalently linking the ASGPR ligand moiety (X) to Y.

[0038] In some embodiments, the ASGPR-binding compound is multivalent (e.g., in Formula (I), n is 2 or more, such that the ASGPR-binding compound comprises two or more ASGPR ligand-binding moieties (X), each covalently attached to a moiety of interest (Y) via a branched linker (e.g., L is a branched linker). In some embodiments, the ASGPR-binding compound is bivalent (e.g., in Formula 1, n is 2). In some embodiments, the ASGPR-binding compound is trivalent (e.g., in Formula 1, n is 3). In some embodiments, each branch of the branched linker is linked to a respective X to a branch point of the linker. In some embodiments, each branch of the linker comprises 14 to 50 consecutive atoms, e.g., 14 to 40, 14 to 30, or 14 to 20 atoms. In some embodiments, each branch of the linker comprises a linear linker of 20 or more consecutive atoms. In some embodiments, the linker comprises 12 or more consecutive atoms covalently bonding the branch point of L to the moiety of interest (Y), e.g., a linear linker of 15 or more, 20 or more, 30 or more, or even more consecutive atoms covalently bonding the branch point of L to Y.

[0039] ASGPR ligand moieties that can be adapted for use in the conjugates of the present disclosure are described in WO / 2023288033, filed July 14, 2022, the entire disclosure of which is incorporated herein by reference. Exemplary ASGPR ligand moieties are described as follows:

[0040] In some embodiments, the ASGPR ligand portion of the bifunctional molecule (e.g., X n -L or (XL) n , and other formulas described herein) have an affinity (K d ) specifically binds to ASGPR. The terms "bind," "bind," "specifically bind," or "specifically bind to" are used interchangeably.

[0041] In some embodiments of the conjugates described herein, X is a group represented by formula (II): [ka] an asialoglycoprotein receptor (ASGPR) binding portion of the formula: R 1 -Z 1 selected from -*, -H, -OH, optionally substituted (C1-C6)alkyl, -OCH3, -OCH2CH=CH, optionally substituted -S-(C1-C6)alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted -S-aryl, and optionally substituted -S-heteroaryl; R 2 -Z 1 -*, -NHCOCH3, -NHCOCF3, -NHCOCH2CF3, -OH, -NHR, and optionally substituted triazole; R 6 -Z 1 -*, -OH, -OR, optionally substituted (C1-C6) alkyl, -OC(O)R, -C(O)NHR, -NR xx R yy , optionally substituted aryl, optionally substituted heteroaryl, -NHCOR, and -NRCOR; each R is independently an optionally substituted (C1-C6) alkyl, an optionally substituted aryl, or an optionally substituted heteroaryl; R xx and R yy are independently H, optionally substituted (C1-C6) alkyl, or R xx and R yy can cyclize to form an optionally substituted heterocyclyl; In the formula, R 1 , R 2 , and R 6 One of them is -Z 1 -*, where "*" represents the connection point between Z1 and the linker (L), R3 and R 4 are each independently H, or a pro-moiety, or R 3 and R 4 are linked in a ring to form a pro-moiety, R 11 is H or a bridging moiety connecting the 5-carbon to the 1-carbon of the ring, Z 1 -Z 11 -, -Z 11 -A 1 -, -A 2 -, -NR 21 CO-, -CONR 21 -, -NR 21 SO2-, -SO2NR 21 -, -NR 21 C(=O)NR 21 -, and -NR 21 C(=S)NR 21 - a linking moiety selected from -Z 11 - is -O-, -S-, -N(R 21 )-, or -C(R 22 )2, where R 1 Ga-Z 1 -*, Z 1 Ga-Z 11 -Z if 11 - is not -O-, -A 1 -and-A 2 - is an optionally substituted heterocyclylene, an optionally substituted arylene, or an optionally substituted heteroarylene; Each R 21 are independently selected from H, optionally substituted (C1-C6) alkyl, -COR, and optionally substituted heteroaryl; Each R 22 are independently selected from H, halogen, and optionally substituted (C1-C6) alkyl.

[0042] In some embodiments, -LY is [ka] wherein R Y teeth, [ka] is.

[0043] In some embodiments, X is a group represented by formula (a-II): [ka] It is expressed as:

[0044] In some embodiments, R 1 -Z 1 In some embodiments, R is -Z 1 -*, -H, or n-propyl.

[0045] In some embodiments, R 2 is Z 1 -* or -NHCOCH3.

[0046] In some embodiments, R 3 and R 4 are -H, respectively.

[0047] In some embodiments, L contains 10 to 60 consecutive branched or linear atoms.

[0048] In some embodiments, L is a group of formula (IIb'): [ka] wherein: L 1 ~L 6 are each independently a linking moiety, and together they form Z 1 and Y, forming a linear or branched linker therebetween; a, b, c, d, and e are each independently 1, 2, 3, 4, or 5; ** is Z 1X's L via 1 represents the attachment point to *** represents the attachment point to Y.

[0049] In some embodiments, L is a group of formula (IIb'): [ka] wherein: n is 1, 2, or 3; L 1 ~L 6 are each independently a linking moiety, and together they form Z 1 and Y, forming a linear or branched linker therebetween; a, b, c, d, and e are each independently 1, 2, 3, 4, or 5; ** is Z 1 X's L via 1 represents the attachment point to *** represents the attachment point to Y.

[0050] In some embodiments, each L 1 ~L 5 -C 1~20 -Alkylene-, -NHC(O)-C 1~6 -Alkylene-, -C(O)NH-C 1~6 -Alkylene-, -NH-C 1~6 -Alkylene-, -NHC(O)NH-C 1~6 -Alkylene-, -NHC(S)NH-C 1~6 -Alkylene-, -C 1~6 -Alkylene-NHC(O)-, -C 1~6 -Alkylene-C(O)NH-, -C 1~6 -Alkylene-NH-, -C 1~6 -Alkylene-NHC(O)NH-, -C 1~6 -Alkylene-NHC(S)NH-, -O(CH2) p -, -(OCH2CH2) pand independently comprise one or more linking moieties independently selected from -, -NHC(O)-, -C(O)NH-, -NHS(O)2-, -S(O)2NH-, -C(O)-, -S(O)2-, -O-, -S-, monocyclic heteroaryl, monocyclic aryl, monocyclic heterocycle, monocyclic carbocycle, an amino acid residue, -NH-, and -NMe-, wherein L 1 ~L 5 are each independently optionally substituted with 1 to 5 halo; each p is independently 1 to 50; L 6 -C 1~20 -Alkylene-, -NR 16 C(O)-C 1~6 -Alkylene-, -C(O)NR 16 -C 1~6 -Alkylene-, -NR 16 -C 1~6 -Alkylene-, -NR 16 C(O)NR 16 -C 1~6 -Alkylene-, -NR 16 C(S)NR 16 -C 1~6 -Alkylene-, -C 1~6 -Alkylene-NR 16 C(O)-, -C 1~6 -Alkylene-C(O)NR 16 -, -C 1~6 -Alkylene-NR 16 -, -C 1~6 -Alkylene-NR 16 C(O)NR 16 -, -C 1~6 -Alkylene-NR 16 C(S)NR 16 -, -O(CH2) p -, -(OCH2CH2) p -, -NR 16 C(O)-, -C(O)NR 16 a linking group comprising one or more linking moieties independently selected from -, -NHS(O)2-, -S(O)2NH-, -C(O)-, -S(O)2-, -O-, -S-, monocyclic heteroaryl, monocyclic aryl, monocyclic heterocycle, an amino acid residue, or -NR16-; Each R 16are independently -H, optionally substituted (C1-C6) alkyl, optionally substituted aryl, optionally substituted monocyclic heteroaryl, or monocyclic heteroaryl.

[0051] In some embodiments, each L 1 ~L 5 -C 1~20 -Alkylene-, -NHC(O)-C 1~6 -Alkylene-, -C(O)NH-C 1~6 -Alkylene-, -NH-C 1~6 -Alkylene-, -NHC(O)NH-C 1~6 -Alkylene-, -NHC(S)NH-C 1~6 -Alkylene-, -C 1~6 -Alkylene-NHC(O)-, -C 1~6 -Alkylene-C( )NH-, -C 1~6 -Alkylene-NH-, -C 1~6 -Alkylene-NHC(O)NH-, -C 1~6 -Alkylene-NHC(S)NH-, -O(CH2) p -, -(OCH2CH2) p -, -NHC(O)-, -C(O)NH-, -NHS(O)2-, -S(O)2NH-, -C(O)-, -S(O)2-, -O-, -S-, monocyclic heteroaryl, monocyclic aryl, monocyclic heterocycle, monocyclic carbocycle, an amino acid residue, -NH-, and -NMe-; 1 ~L 5 are each independently optionally substituted with 1 to 5 halo; each p is independently 1 to 50; L 6 teeth, [ka] where: R z teeth, [ka] is.

[0052] 1. Linked ASGPR Ligand Moiety In some embodiments, the ASGPR binding moiety (X) of the compounds of the present disclosure has formula (Iia): [ka] where R 2 , R 3 , R 4 , R 6 and Z 1 is as defined herein. In some embodiments of Formula (Iia), R 6 is selected from -OH, -OC(O)R, and -C(O)NHR, and R 2 is selected from -NHCOCH3, -NHCOCF3, and -NHCOCH2CF3.

[0053] In some embodiments of Formula (II), Z 1 is in the β configuration, and the formula (Iia-1) [ka] It can be described as:

[0054] In some embodiments of Formula (II), Z 1 is in the α configuration, and the formula (Iia-2) [ka] It can be described as:

[0055] In certain embodiments of formula (Iia), (Iia-1), or (Iia-2), Z 1 -Z 11 -A 1 - in which A 1- is an optionally substituted arylene or an optionally substituted heteroarylene. In certain embodiments, A is an optionally substituted heteroarylene. In certain embodiments, the heteroarylene is a 5- or 6-membered heteroarylene. In certain embodiments, the heteroarylene is a 5-membered heteroarylene. In certain embodiments, the 5-membered heteroarylene is a triazole. In certain embodiments, the triazole is a 1,2,3-triazole moiety.

[0056] In some embodiments, the ASGPR binding moiety (X) of the compounds of the present disclosure has formula (IIIa) or (IIIb): [ka] where: -Z 11 - is -O-, -S-, -N(R 21 )-, or -C(R 22 )2-, wherein each R 22 are independently selected from H, halogen (e.g., F), and optionally substituted (C1-C6) alkyl; R 21 is H or optionally substituted (C1-C6) alkyl; -A 1 - is arylene, substituted arylene, heteroarylene, or substituted heteroarylene.

[0057] In some embodiments of formula (IIIa) or (IIIb), Z 11 is -S-.

[0058] In some embodiments, Z 11 is -C(R 22 )2-. In some embodiments, Z 11 is -CH2-.

[0059] In certain embodiments, Z 11 is -C(R 22 )2, wherein at least one R 22is H. In certain embodiments, both R 22 is H. In certain embodiments, Z 11 is —O—. In certain embodiments, Z 11 In certain embodiments, Z is -S-. 11 is -N(R 21 ) where R 21 is H or (C1-C3) alkyl.

[0060] In certain embodiments, -A 1 - is a triazole.

[0061] In certain embodiments, Z 1 is -C(R 22 ) 2-triazole. In certain embodiments, Z 1 teeth, [ka] In certain embodiments, Z 1 teeth, [ka] is.

[0062] In certain embodiments of formula (Iia), (Iia-1), or (Iia-2), Z 1 is Z 11 In certain embodiments, Z 11 is -C(R 22 )2. In certain embodiments, at least one R 22 is H. In certain embodiments, both R 22 is H, Z 11 is -CH2-. In some cases, Z 11 is —O—. In certain embodiments, Z 11 is -S-. In certain other cases, Z 11 is -N(R 21 ) where R 21 is H or (C1-C3) alkyl.

[0063] In certain embodiments of formula (Iia), (Iia-1), or (Iia-2), Z 1 is a monocyclic 5- or 6-membered heteroaryl or aryl. In certain embodiments, Z 1 teeth, [ka] In certain embodiments, Z 1 teeth, [ka] is.

[0064] In certain embodiments of formula (Iia), (Iia-1), or (Iia-2), Z 1 -O-, -S-, -C(R 22 )2-, -NR 21 -,-CONR 21 - and [ka] is selected from: X 1 is O or S, t is 0 or 1, R 21 and each R 23 is 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.

[0065] In certain embodiments of formula (Iia), (Iia-1), or (Iia-2), Z 1 is an optionally substituted (C1-C6) alkyl. Z 1 In certain embodiments, alkyl is methyl. 1 In certain embodiments, alkyl is ethyl.1 In certain embodiments, alkyl is propyl. 1 In certain embodiments, alkyl is butyl. 1 In certain embodiments, alkyl is pentyl. 1 In certain embodiments, alkyl is hexyl.

[0066] In certain embodiments, the ASGPR binding moiety (X) of formula (Iia-1) is selected from one of the following structures: [ka]

[0067] In some embodiments of formula (Iia-2), Z 1 is in the β configuration, and X is represented by the formula (IIIb-2) [ka] wherein: -A 1 - is arylene, substituted arylene, heteroarylene, or substituted heteroarylene.

[0068] In some embodiments of formula (IIIb-2), A 1 In some embodiments of formula (IIIb-2), X is a group represented by formula (X A -4).

[0069] In some embodiments of formula (Iia-1), Z 1 is in the alpha configuration at the 1 carbon of the galactosamine ring. In some embodiments of formula (Iia-1), Z 1 is S, and each X is a function of the formula (X A In some embodiments of formula (Iia-1), each X is of formula (X A In some embodiments of formula (Iia-1), each X is of formula (X AIn some embodiments of formula (Iia-1), each X is of formula (X A In some embodiments of formula (Iia-1), each X is of formula (X A -5).

[0070] In certain embodiments, the compound of formula (Iia-2) is selected from one of the following structures: [ka]

[0071] In some embodiments of formula (Iia-2), each X is a group represented by the formula (X B -1).

[0072] In some embodiments of formula (Iia-2), each X is a group represented by the formula (X B -2).

[0073] In some embodiments of formula (Iia-2), each X is a group represented by the formula (X B -3).

[0074] In some embodiments of formula (Iia-2), each X is a group represented by the formula (X B -4).

[0075] In some embodiments of formula (Iia-2), Z1 is in the α configuration and X is a group represented by formula (IIIb-1): [ka] wherein -A 1 - is arylene, substituted arylene, heteroarylene, or substituted heteroarylene.

[0076] In certain embodiments of Formula (IIIb-1), A 1is an optionally substituted heteroarylene. In certain embodiments, the heteroarylene is a 5- or 6-membered heteroarylene. In certain embodiments, the heteroarylene is a 5-membered heteroarylene. In certain embodiments, the 5-membered heteroarylene is a triazole. In certain embodiments, the triazole is a 1,2,3-triazole moiety.

[0077] In certain embodiments, X in formula (IIIb-1) is selected from one of the following structures: [ka]

[0078] In some embodiments of formula (IIIb-1), each X is a group represented by the formula (X C -1).

[0079] In some embodiments of formula (IIIb-1), each X is a group represented by the formula (X C -2).

[0080] Exemplary ligand moieties that bind to ASGPR and their synthons that can be utilized in compounds of the present disclosure are shown in Tables 1-5. In certain embodiments, the compound of formula (Iia) is a compound shown in Table 1. [Table 1]

[0081] In some embodiments of any one of X1 to X5.1, Z 1 is in the α-configuration, so that the ASGPR binding moieties X1 to X5.1 are represented by the formula (Iia-2): [ka] is derived from

[0082] Two linked ASGPR ligand moieties In some embodiments, the ASGPR binding moiety (X) is linked to the 2-position of the sugar analog. In some embodiments, the ASGPR binding moiety (X) has a reduced ring carbon at the 1-position relative to the galactosamine-derived sugar. In some embodiments, the ASGPR binding moiety (X) of the bifunctional molecules of the present disclosure has formula (Iib): [ka] where R 1 , R 3 , R 4 , R 6 , R 11 and Z 1 is as defined herein.

[0083] In some embodiments, the ASGPR binding moiety (X) of the compounds of the present disclosure has the formula (Iib'): [ka] where R 3 ~R 4 , R 6 , and Z 1 is as defined herein.

[0084] In some embodiments, the ASGPR binding moiety (X) of the compounds of the present disclosure has the formula (Iva): [ka] where R 1 , R 11 , and Z 1 is as defined herein.

[0085] In some embodiments of formula (Iib), (Iib') or (Iva), Z 1 is an optionally substituted -(C(R 22 )2) q heteroarylene, and [ka] wherein q is 0 or 1.

[0086] In some embodiments of formula (Iib), (Iib') or (Iva), Z 1 is an optionally substituted -(C(R 22 )2) q -triazole, where q is 0 or 1.

[0087] In some embodiments of formula (Iib), (Iib') or (Iva), Z 1 teeth, [ka] In some embodiments, Z 1 teeth, [ka] is.

[0088] In some embodiments of formula (Iib), (Iib') or (Iva), Z 1 teeth, [ka] where R 23 is H or C (1~3) - alkyl.

[0089] In some embodiments of formula (Iib), (Iib') or (Iva), Z 1 is -NR 23 CO—, where R 23 is H or C (1~3) - alkyl.

[0090] In some embodiments of formula (Iib), (Iib') or (Iva), Z 1 is an optionally substituted -(C(R 22 )2) q heteroaryl, and [ka] wherein q is 0 or 1.

[0091] In certain embodiments of formula (Iib), (Iib') or (Iva), Z 1 is an optionally substituted -(C(R 22 )2) q -triazole, where q is 0 or 1. In certain embodiments, Z 1 teeth, [ka] is.

[0092] In certain cases of formula (Iib), (Iib') or (Iva), Z 1 teeth, [ka] where R 23 is H or C (1~3) - alkyl.

[0093] In certain cases of formula (Iib), (Iib') or (Iva), Z 1 is -NR 23 CO—, where R 23 is H or C (1~3) - alkyl.

[0094] In certain embodiments of formula (Iib), (Iib') or (Iva), Z 1 is a monocyclic 5- or 6-membered heteroarylene or arylene. In certain embodiments, Z 1 teeth, [ka] is.

[0095] In certain embodiments of formula (Iib), (Iib'), or (Iva), Z 1-O-, -S-, -C(R 22 )2-, -NR 21 -,-CONR 21 - and [ka] is selected from: X 1 is O or S, t is 0 or 1, R 21 and each R 23 is 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.

[0096] In certain embodiments, the compound of formula (Iib), (Iib') or (Iva) is selected from one of the following structures: [ka] In the formula, R 1A are independently H or (C 1~3 ) alkyl.

[0097] In some embodiments, the ASGPR binding moiety (X) of the compounds of the present disclosure has formula (Ivb) or (Ivc): [ka] where: -Z 11 - is -O-, -S-, -N(R 21 )-, or -C(R 22 )2, -A 1 -and-A 2 - is an optionally substituted arylene or an optionally substituted heteroarylene; Each R 21are independently selected from H and optionally substituted (C1-C6) alkyl; Each R 22 is independently selected from H, halogen (e.g., F), and optionally substituted (C1-C6) alkyl. In some embodiments of formula (Ivb) or (Ivc), R 1 is H.

[0098] In some embodiments, R 2 -Z 1 -* and R 11 is a group of formula -CH2O- that bridges (i.e., cyclically joins) the 1-carbon atom of the sugar ring.

[0099] In some embodiments, -Z 1 -* or -Z 1 -L- is [ka] Includes.

[0100] In certain embodiments of Formula (Iib), R 11 is H and the compounds are those in Table 2. [Table 2]

[0101] In certain embodiments, the compound of formula (Iib) is a compound shown in Table 3. In certain embodiments, the compound of formula (Iib), C1 (i.e., R 1 In certain embodiments, the configuration in the compound of formula (Iib), C1 (i.e., R 1 ) is configured as β. [Table 3-1] [Table 3-2] [Table 3-3]

[0102] In certain embodiments, the compound of formula (Id') is a compound shown in Table 4. [Table 4]

[0103] In some embodiments, the ASGPR binding moiety (X) of a compound of the present disclosure has formula (Ivb-1) or (Ivc-1): [ka] where R 11 is the bridging moiety connecting the 5 carbon to the 1 carbon.

[0104] In some embodiments of formula (Ivb), or (Ivb-1), Z 11 is -C(R 22 )2. In certain embodiments, at least one R 22 is H. In certain embodiments, both R 22 is H. In certain embodiments, Z 11 is —O—. In certain embodiments, Z 11 is -S-. In certain embodiments, Z 11 is -N(R 21 ) where R 21 is H or (C1-C3) alkyl.

[0105] In certain embodiments of formula (Ivb), (Ivc), (Ivb-1) or (Ivc-1), -A 1 -and-A 2 Each - is independently an optionally substituted heteroarylene. In certain embodiments, the heteroarylene is a 5- or 6-membered heteroarylene. In certain embodiments, the heteroarylene is a 5-membered heteroarylene. In certain embodiments, the heteroarylene is a 6-membered heteroarylene.

[0106] In some embodiments of formula (Ivb) or (Ivb-1), A 1 The ring is a 5-membered heteroarylene selected from triazole, thiadiazole, thiophene, oxazole, isoxazole, isothiazole, thiazole, oxadiazole, and furan. 1 The ring is a 6-membered heteroarylene selected from pyridine, pyrimidine, pyridazine, pyrazine, and triazine. 1 The ring is a triazole. 1 The ring is pyridine. 1 The ring is pyrimidine. 1 In certain embodiments, the ring is a thiadiazole. 1 The ring is a 5- or 6-membered arylene or heteroarylene, further substituted with one or more substituents. 1 The ring is further substituted with one or more substituents selected from halogen, (C1-C6) alkyl, and substituted (C1-C6) alkyl (eg, CF3).

[0107] In some embodiments of any one of formulas (Ivc) or (Ivc-1), A 2 The ring is a 5-membered heteroarylene selected from triazole, thiadiazole, thiophene, oxazole, isoxazole, isothiazole, thiazole, oxadiazole, and furan. 2 The ring is a 6-membered heteroarylene selected from pyridine, pyrimidine, pyridazine, pyrazine, and triazine. 2 The ring is a triazole. 2 The ring is pyridine. 2 The ring is pyrimidine. 2 In certain embodiments, the ring is a thiadiazole. 2The ring is a 5- or 6-membered arylene or heteroarylene, further substituted with one or more substituents. 2 The ring is further substituted with one or more substituents selected from halogen, (C1-C6) alkyl, and substituted (C1-C6) alkyl (eg, CF3).

[0108] In certain embodiments of formula (Ivb) or (Ivb-1), -Z 11 -A 1 - is a monocyclic 5- or 6-membered heteroarylene of one of the following structures: [ka]

[0109] In certain embodiments of formula (Ivc) or (Ivc-1), -A 2 - is a monocyclic 5- or 6-membered heteroarylene of the structure: [ka]

[0110] Specific -Z 11 -A 1 It is understood that a variety of substituents can be utilized to connect the - group to the adjacent linker. In certain embodiments of formula (Ivb) or (Ivb-1), -Z 11 -A 1 - is a monocyclic 5- or 6-membered heteroarylene attached to a linking moiety as shown in one of the following structures: [ka]

[0111] In certain embodiments of formula (Ivc) or (Ivc-1), -Z 11 -A 1 - is a monocyclic 5- or 6-membered heteroarylene attached to a linking moiety as shown in one of the following structures: [ka]

[0112] In some embodiments of the compound of formula (Iib), or (Iva)-(Ivc), R 1 is H, and as a result, the compounds of formula (Iib) or (Iva) to (Ivc) have no substituent other than hydrogen at the 1-position of the sugar ring.

[0113] In some embodiments, the compound of Formula (Iib) is any one of Formulas (Ivd)-(Ivg): [ka] In the formula, A 1 Ring and A 2 Ring, R 6 , R 4 , R 3 , R 11 , and R 21 is as defined herein.

[0114] In some embodiments of any one of formulas (Ivd)-(Ivg), A 1 The ring is a 5- or 6-membered arylene or heteroarylene. 1 The ring is a 5-membered heteroarylene selected from triazole, thiadiazole, thiophene, oxazole, isoxazole, isothiazole, thiazole, oxadiazole, imidazole, and furan. 1 The ring is a 6-membered heteroarylene selected from pyridine, pyrimidine, pyridazine, pyrazine, and triazine. 1 The ring is a triazole. 1 The ring is pyridine. 1 The ring is pyrimidine. 1 In certain embodiments, the ring is a thiadiazole. 1The ring is pyrazine. 1 The ring is a 5- or 6-membered arylene or heteroarylene, further substituted with one or more substituents. 1 The ring is further substituted with one or more substituents selected from halogen, (C1-C6) alkyl, and substituted (C1-C6) alkyl (eg, CF3).

[0115] In some embodiments of any one of formulas (Ivd)-(Ivg), A 1 The ring is a 5- or 6-membered arylene or heteroarylene. 2 The ring is a 5-membered heteroarylene selected from triazole, thiadiazole, thiophene, oxazole, isoxazole, isothiazole, thiazole, oxadiazole, and furan. 2 The ring is a 6-membered heteroarylene selected from pyridine, pyrimidine, pyridazine, pyrazine, and triazine. 2 The ring is a triazole. 2 The ring is pyridine. 2 The ring is pyrimidine. 2 In certain embodiments, the ring is a thiadiazole. 2 The ring is a 5- or 6-membered arylene or heteroarylene, further substituted with one or more substituents. 2 The ring is further substituted with one or more substituents selected from halogen, (C1-C6) alkyl, and substituted (C1-C6) alkyl (eg, CF3).

[0116] In some embodiments of any one of formulas (Ivd)-(Ivg), A 1 Ring or A 2 There is no ring.

[0117] In some embodiments of any one of formulas (Ivd)-(Ivg), A 1 Ring or A 2 The ring is phenylene or substituted phenylene.

[0118] In some embodiments of Formula (Ivd), A 2 The ring is a 5- or 6-membered heteroarylene. In certain cases of formula (Ivd), A 2 The ring is a 5-membered heteroarylene. In certain embodiments of Formula (Ivd), A 2 In certain embodiments of (Ivd), A 2 There is no ring.

[0119] In some embodiments of Formula (Ive), A 1 The ring is a 5- or 6-membered heteroarylene, and R 21 is H. In certain embodiments of Formula (Ive), the A ring is triazole. In certain cases of Formula (Ive), A 1 In certain cases of formula (Ive), A 1 In certain cases of formula (Ive), A 1 In some embodiments of Formula (Ive), A 1 The ring is absent, and R 21 is H or optionally substituted acyl. In certain embodiments, R 21 is —COCH. In certain embodiments, R 21 is H.

[0120] In some embodiments of Formula (Ivf), A 1 The ring is a 5- or 6-membered heteroarylene. In certain cases of formula (Ivf), A 1 The ring is a 5-membered heteroarylene. In certain embodiments of Formula (Ivf), A 1 In certain embodiments of (Ivf), A 1 There is no ring.

[0121] In some embodiments of Formula (Ivg), A 2 The ring is a 5- or 6-membered heteroarylene. In certain cases of formula (Ivg), A 2 The ring is a 5-membered heteroarylene. In certain embodiments of Formula (Ivg), A 2 The ring is a triazole. (In certain embodiments of Ivdg, A 2 There is no ring.

[0122] In certain embodiments, the ASGPR binding moiety (X) of the compounds of the present disclosure is represented by formula (Ivh)-(Ivk): [ka] wherein: R 6 , R 4 , R 3 , and R 21 is as defined herein; Y 1 ~Y 3 are each independently N or CR 25 and R 24 and R 25 are H, optionally substituted C (1~6) -alkyl, optionally substituted fluoroalkyl, and halogen.

[0123] In certain embodiments, the ASGPR binding moiety (X) of the compounds of the present disclosure is represented by the formula ((Ivl)-(Ivm): [ka] wherein: R 6 , R 4 , R 3 , and R 21 is as defined herein; Y 1 ~Y 3 are each independently N or CR 25and Y 4 is N or CR 24 and Y 5 is S, O, or NH, R 24 and R 25 are H, optionally substituted C (1~6) -alkyl, optionally substituted fluoroalkyl, and halogen.

[0124] In some embodiments of Formula (Ivi), Y 1 ~Y 3 At least one of is N.

[0125] In certain embodiments, Y 1 ~Y 3 At least two of the are N.

[0126] In certain embodiments, Y 1 and Y 4 is N.

[0127] In certain embodiments, Y 1 and Y 3 is N and Y 2 is CR 25 is.

[0128] In certain embodiments, Y 1 and Y 2 is N and Y 3 is CR 25 is.

[0129] In certain embodiments, Y 1 and Y 2 is CR 25 and Y 3 is N.

[0130] In certain embodiments of any one of Formulas (Ivd)-(Ivk), or (Ivd)-(Ivm), R 6 is H.

[0131] In some embodiments of any one of Formulas (Ivd)-(Ivk), or (Ivd)-(Ivm), R 4 and R 3 are each H. In certain embodiments, R 4 ~R 3 At least one of R is a pro-moiety. 4 and R 3 are cyclically linked to form a promoiety (eg, as described herein).

[0132] In some embodiments, the compound of formula (Ivi) has the formula (Ivi-1): [ka] It is of

[0133] In the formula, R 24 and R 25 is independently selected from H, halogen, (C1-C6) alkyl, and substituted (C1-C6) alkyl (e.g., CF3). In some embodiments of Formulas (Ivi)-(Ivi-1), R 25 is H. In certain embodiments, R 25 is C (1~3) -Alkyl, or C (1~3) -fluoroalkyl. In certain embodiments, fluoroalkyl is CF. In some embodiments of Formula (Ivi) or (Ivi-1), R 24 is H. In certain embodiments, R 24 is C (1~3) -Alkyl, or C (1~3) -fluoroalkyl. In certain embodiments, fluoroalkyl is CF.

[0134] In some embodiments, the compound of formula (Ivi-1) has the formula (XD): [ka] It is of the type.

[0135] In some embodiments, the compound of formula (Ivk-1) is a compound of formula (X E ) [ka] It is of the type.

[0136] In certain embodiments, the compound of formula (Ivl) has the formula (Ivl-1): [ka] wherein: R 6 , R 4 , R 3 , and R 21 is as defined herein; Y 1 ~Y 4 are each independently N or CR 25 and Y 5 is S, O, or NH, Each R 25 is H, optionally substituted C (1~6) -alkyl, optionally substituted fluoroalkyl, and halogen.

[0137] In certain embodiments, each R 25 is H.

[0138] In certain embodiments, the ASGPR binding moiety (X) of the compounds of the present disclosure can be described by one of the following structures: [ka]

[0139] In certain embodiments, the ASGPR binding moiety (X) of the compounds of the present disclosure can be described by one of the following structures: [ka]

[0140] In certain embodiments, the ASGPR binding moiety (X) of the compounds of the present disclosure can be described by one of the following structures: [ka] In certain embodiments, the ASGPR binding moiety (X) of the compounds of the present disclosure can be described by one of the following structures: [ka]

[0141] In certain embodiments of Formula (Iib), R 1 R 3 , R 4 , and R 11 is H, R 6 is OH, [ka] In the formula, Z 1 is -NH-, -CH2-, -S- or -O-.

[0142] In certain embodiments of Formula (Iib′), R 3 , R 4 is H, R 6 is OH, [ka] In the formula, Z 1 is -NH-, -CH2-, -S-, -O-, triazole, e.g. [ka] is.

[0143] 6-linked ASGPR ligand moiety In some embodiments, the ASGPR binding moiety (X) is linked to position 6 of the sugar analog. In some embodiments, the ASGPR binding moiety (X) has a reduced ring carbon at position 1 relative to the galactosamine-derived sugar.

[0144] In some embodiments, the ASGPR binding moiety (X) of the compounds of the present disclosure has formula (Iic): [ka] where R 1 ~R 4 , and Z 1 is as defined herein.

[0145] In certain embodiments of Formula (Iic), Z1 is selected from -O-, -S-, -CONR21-, and optionally substituted -(C(R22)2)q-heteroarylene, where q is 0 or 1. In certain other cases, Z1 is an optionally substituted -(C(R22)2)q-triazole, where q is 0 or 1. In certain embodiments, Z1 is -O-. 1 teeth, [ka] is.

[0146] In certain embodiments of Formula (Iic), Z 1 -Z 11 -A 1 - in which -A 1 - is an optionally substituted -A 1 - or optionally substituted arylene. In certain embodiments, -A 1- is an optionally substituted heteroarylene. In certain embodiments, the heteroarylene is a 5- or 6-membered heteroarylene. In certain embodiments, the heteroarylene is a 5-membered heteroarylene. In certain embodiments, the 5-membered heteroarylene is a triazole. In certain embodiments, the triazole is a 1,2,3-triazole moiety. In certain embodiments, Z 11 is -C(R 22 )2. In certain embodiments, at least one R 22 is H. In certain embodiments, both R 22 is H. In certain cases, Z 11 is —O—. In certain embodiments, Z 11 is -S-. In certain other cases, Z 11 is -N(R 21 ) where R 21 is H or (C1-C3) alkyl. In certain embodiments, Z 1 is -C(R 22 ) 2-triazole. In certain embodiments, Z 1 teeth, [ka] is.

[0147] In certain embodiments of Formula (Iic), Z 1 is Z 11 In certain embodiments, Z 11 is -C(R 22 )2. In certain embodiments, at least one R 22 is H. In certain embodiments, both R 22 is H, Z 11 is -CH2-. In some cases, Z 11 is —O—. In certain embodiments, Z 11 is -S-. In certain other cases, Z 11 is -N(R 21 ) where R 21is H or (C1-C3) alkyl.

[0148] In certain embodiments of Formula (Iic), Z 1 is a monocyclic 5- or 6-membered heteroarylene or arylene. In certain embodiments, Z 1 teeth, [ka] is.

[0149] In certain embodiments of Formula (Iic), Z1 is -O-, -S-, -C(R22)2-, -N(R21)-CON(R 21 )-, and [ka] is selected from: X 1 is O or S, t is 0 or 1, R 21 and each R 23 is 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.

[0150] In certain embodiments, the compound of formula (Iic) has the following structure: [ka]

[0151] In certain embodiments, the compound of formula (Iic) has the following structure: [ka]

[0152] In certain embodiments of Formula (Iic), R 11 is H and the compounds are those in Table 5. [Table 5]

[0153] In some embodiments, the ASGPR binding moiety (X) of the compounds of the present disclosure has the formula (Iid): [ka] where: R 6 , R 4 , R 3 , and Z 1 is as defined herein; Y 6 and Y 5 are -O-, -S-, and NR, respectively. 21 - and -C(R 22 )2 are independently selected from R 21 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 (Iid), Y 5 is attached to the sugar ring via an α-configuration. In some embodiments of formula (Iid), Y 5 is connected to the sugar ring via a β configuration.

[0154] In some embodiments, the ASGPR binding moiety (X) of the compounds of the present disclosure has the formula (Iid'): [ka] where: R 6 , R 4 , R 3, and Z 1 is as defined herein; Y 5 and Y 6 are -O-, -S-, and NR, respectively. 21 - and -C(R 22 )2 are independently selected from R 21 is H, optionally substituted (C1-C6) alkyl, and -C(O)R 22 is selected from Each R 22 is H, halogen, and optionally substituted (C 1 ~C 6 ) alkyl; Ring B is a 5- or 6-membered optionally substituted cyclic group.

[0155] In some embodiments of formula (Iid)-(Iid′), Y 5 is O. In certain embodiments, Y 5 is S. In certain embodiments, Y 5 is -NR 21 In certain embodiments, Y 5 is -C(R 22 )2, and each R 22 is H.

[0156] In some embodiments of formula (Iid)-(Iid′), Y 6 is -NR 21 -, wherein R 21 is H. In certain embodiments, Y 6 is -NR 21 -, wherein R 21 is -C(O)R 22 In certain embodiments, R 22 is methyl.

[0157] In some embodiments of Formula (Iid)-(Iid'), the B ring is a 5-membered or 6-membered heterocycle. In certain embodiments, the B ring is a 5-membered heterocycle. In certain embodiments, the B ring is a 6-membered heterocycle.

[0158] In some embodiments of formulas (Iid)-(Iid′), Z 1 is Z 11 wherein Z 11 -O-, -S-, NR 21 - and -C(R 22 )2. In certain embodiments, Z 1 is —O—. In certain embodiments, Z 1 is -S-. In certain embodiments, Z 1 is NR 21 wherein R21 is H. In certain embodiments, Z 1 is -C(R 22 )2, wherein each R 22 is H.

[0159] In some embodiments of formulas (Iid)-(Iid′), Z 1 is an arbitrarily substituted Z 11 -heteroarylene or optionally substituted Z 11 -arylene. In some embodiments, Z 1 is CH2-heteroarylene or CH2-arylene. In some embodiments of Formula (Iid)-(Iid'), Z 1 is an optionally substituted amide. In some embodiments of Formula (Iid)-(Iid'), Z 1 is an optionally substituted sulfonamide. In some embodiments of Formula (Iid)-(Iid'), Z 1 is an optionally substituted urea or an optionally substituted thiourea.

[0160] In some embodiments, the compound of Formula (Iid)-(Iid') has one of the following structures: [ka]

[0161] In certain embodiments of any one of formulas (Iia), (Iib), or (Iid), R6 is OH. In certain other cases, R 6 is —OC(O)R. In certain embodiments, R 6 is —C(O)NHR, where R is an optionally substituted alkyl. In certain embodiments, R terminates in an alkenyl or alkynyl group. In certain other cases, R 6 is an optionally substituted triazole. In certain embodiments, the triazole has the structure [ka] It is of the type.

[0162] In certain embodiments of formula (Iia) and (Iic), R 2 is —NHCOCH. In certain other embodiments, R 2 is -NHCOCF. In certain other embodiments, R 2 is —NHCOCH2CF3. In certain embodiments, R 2 is —OH. In certain other cases, R 2 is an optionally substituted triazole. In certain embodiments, the triazole has the structure [ka] It is of the type.

[0163] In certain embodiments, R 6 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. In certain embodiments, the substituent on the triazole moiety is optionally substituted (C 1~6 ) alkyl, optionally substituted (C 1~6) alkoxy, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted alkaryl, and optionally substituted alkylheteroaryl. It will be understood that any convenient substituent can be included in the triazole moiety. See, for example, the triazole moieties disclosed in Mamidayala et al., J. Am. Chem. Soc. 2012, 134, 1978-1981.

[0164] Z 1 , Z 11 , and Z 11 It is understood that the -Ar linking moiety can be considered part of the X group of formula (I). In the ASGPR binding moieties (X) described herein, -Z 1 - can be connected to -L (e.g., in a linker described herein) via various bonds and linking moieties depending on the method of preparation. 1 In some embodiments, the subject compound can be linked to a - moiety. [ka] Selected from -Z 1 -L 1 - includes parts, In the formula, each R 21 is independently selected from H, and optionally substituted (C-C) alkyl, and each R 22 are independently selected from H, halogen (e.g., F), and optionally substituted (C1-C6) alkyl, and o, p, q, r, s, t, u, v, w, x, y, z, and z1 are each independently 0-6.

[0165] In some embodiments, the subject compound is [ka] Selected from -Z 1 -L 1 - includes parts, In the formula, each R 21 is independently selected from H, and optionally substituted (C-C) alkyl, and each R22 are independently selected from H, halogen (e.g., F), and optionally substituted (C1-C6) alkyl, and o, p, q, r, s, t, u, v, w, x, y, z, and z1 are each independently 1 to 6.

[0166] In certain embodiments, Z 1 -L 1 -The group is [ka] and o is 1 or 2.

[0167] In certain embodiments, Z 1 -L 1 -The group is [ka] and each R 22 is H and p is 1 or 2.

[0168] In certain embodiments, Z 1 -L 1 -The group is [ka]

[0169] In certain embodiments, Z 1 -L 1 -The group is [ka] wherein r is 1 to 3.

[0170] In certain embodiments, Z 1 -L 1 -The group is [ka] wherein r is 1 to 3.

[0171] In certain embodiments, Z 1 -L 1 -The group is [ka] wherein s and t each independently represent an integer of 1 to 3.

[0172] In certain embodiments, Z 1 -L 1 -The group is [ka] wherein u is 1 to 3.

[0173] [ka] In the formula, v and w each independently represent an integer of 1 to 3.

[0174] In certain embodiments, Z 1 -L 1 -The group is [ka] In the formula, x is 0 to 3.

[0175] In certain embodiments, Z 1 -L 1 -The group is [ka] where y is 1 to 3.

[0176] In certain embodiments, Z 1 -L 1 -The group is [ka] where R 21 is H and z is 1 to 4.

[0177] In certain embodiments, Z 1 -L 1 -The group is [ka] where R 21 is H, and z1 is 1 to 4.

[0178] In certain embodiments, Z 1 -L 1 -The group is [ka] and each R 22 is H and q is 0 to 3. In certain embodiments, Z 1 -L 1 -The group is [ka]

[0179] In certain embodiments, Z 1 -L 1 -The group is [ka] wherein q is 1 to 3.

[0180] In some embodiments, the subject compound is [ka] Selected from -Z 1 Contains the -L- group.

[0181] In certain embodiments, Z 1 -L 1 -The group is [ka] wherein q is 1 to 3. In certain embodiments, q is 1. In certain embodiments, q is 2. In certain embodiments, q is 3.

[0182] In certain embodiments, Z 1 -L 1 -The group is [ka] is.

[0183] In certain embodiments, Z 1 -L 1 -The group is [ka] is.

[0184] In certain embodiments, Z 1 -L 1 -The group is [ka] is.

[0185] In certain embodiments, -Z 1 -L 1 - includes optionally substituted -NH-heteroarylene-. In certain embodiments, a heteroarylene is triazole. In certain embodiments, a heteroarylene is pyridine. In certain embodiments, a heteroarylene is pyrimidine. In certain embodiments, a heteroarylene is thiadiazole.

[0186] In certain embodiments, -Z 1 -L 1 -teeth, [ka] wherein: Each R 21are independently selected from H, optionally substituted (C-C) alkyl, and optionally substituted acyl; R 24 and R 25 are H, optionally substituted C, (1~6) -alkyl, optionally substituted fluoroalkyl, and halogen.

[0187] In certain embodiments, -Z 1 -L 1 -teeth, [ka] wherein: R 24 and R 25 are H, optionally substituted C, (1~6) - independently selected from alkyl, optionally substituted fluoroalkyl, and halogen; 21 are independently selected from H, optionally substituted (C1-C6) alkyl, and optionally substituted acyl.

[0188] In certain embodiments, R 21 is H. In certain embodiments, R 24 is C (1~3) -Alkyl, or C (1~3) -fluoroalkyl. In certain embodiments, fluoroalkyl is CF. In certain embodiments, R 25 is C (1~3) -Alkyl, or C (1~3) -fluoroalkyl. In certain embodiments, fluoroalkyl is CF.

[0189] In certain embodiments, -Z 1 -L 1 -teeth, [ka]

[0190] It is understood that a variety of substituents and chemistries can be utilized to connect a particular X ligand moiety (e.g., as described herein) to an adjacent linker. In some embodiments, the linking moiety of the linker comprises a triazole derived from a click chemistry bond. In certain embodiments, the ASGPR ligand moiety (X) is attached to the linking moiety as shown in any one of the following structures: [ka]

[0191] In certain embodiments of Formula (Iib), R 1 R 3 , R 4 , and R 11 is H, R 6 is OH, [ka]

[0192] In certain embodiments of Formula (Iib), R 1 R 3 , R 4 , and R 11 is H, R 6 is OH, [ka] In the formula, Z 1 is attached to a linking moiety as shown in one of the following structures: [ka]

[0193] M6PR ligand moiety As summarized above, the M6PR-binding moieties (also referred to as M6PR ligand moieties) of the present disclosure can be linked to a variety of moieties of interest without affecting their specific binding to cell-surface M6PR and its function. The present inventors have demonstrated that M6PR-binding moieties having the specific structures described below provide high-affinity binding to cell-surface M6PR and, when configured via a linker according to the bifunctional compounds of the present disclosure, can exploit the function of cell-surface M6PR in biological systems, such as internalization and / or degradation of target molecules.

[0194] The terms "mannose-6-phosphate receptor" and "M6PR" refer to receptors in the mannose-6-phosphate receptor family. M6PR is a transmembrane glycoprotein receptor that targets enzymes to lysosomes within cells. M6R endogenously transports proteins bearing N-glycans covered with mannose-6-phosphate (M6P) residues to lysosomes and cycles between endosomes, the cell surface, and the Golgi apparatus. See, for example, Ghosh et al., Nat. Rev. Mol. Cell Biol. 2003;4:202-213. The M6PR family includes the cation-independent mannose-6-phosphate receptor (CI-M6PR). CI-M6PR, also known as insulin-like growth factor 2 receptor (IGF2R), is encoded by the IGF2R gene in humans (see, for example, NCBI Reference Sequence: NM_000876.3 and NCBI Gene ID: 3482). CI-M6PR binds to insulin-like growth factor 2 (IGF-2) and mannose-6-phosphate (M6P) tagged proteins. The compounds of the present disclosure can specifically bind to cell surface M6PR, for example, internalized CI-M6PR cell surface receptors. In certain embodiments, the surface CI-M6PR is human CI-M6PR. It is understood that the terms M6PR and CI-M6PR are used interchangeably when referring to the binding properties of the M6PR binding moieties and compounds of the present disclosure.

[0195] Compounds comprising such an M6PR binding moiety (X) (e.g., as described herein) may bind to other receptors, or may bind with lower affinity, e.g., as determined by immunoassays or other assays known in the art. In certain embodiments, X, or a compound described herein comprising such X, specifically binds to cell surface CI-M6PR with an affinity of at least 2, 2.5, 3, 4, or more logs greater than the affinity with which X or the compound binds to another cell surface receptor. In certain embodiments, X, or a compound described herein comprising X, specifically binds to cell surface CI-M6PR with an affinity (K d ) specifically binds to CI-M6PR. In certain embodiments, such binding is characterized by an 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. The terms "bind," "bind," "specifically bind," or "specifically bind to" are used interchangeably.

[0196] The M6PR-binding compounds of the present disclosure include a moiety (X) (e.g., as described herein) that is a D-mannopyranose analog that specifically binds to the cell surface receptor M6PR. M6PR-binding compounds can be monovalent or multivalent (e.g., bivalent or trivalent or higher), with monovalent compounds containing a single M6PR ligand moiety and monovalent compounds containing two or more such moieties.

[0197] α-linked pyranose ring The M6PR binding portion of the compounds of the present disclosure has formula (II″): [ka] and wherein: W is a hydrophilic head group; Z 1 is selected from optionally substituted (C1-C3) alkylene and optionally substituted ethenylene; Z 2 are O, S, NR 21and C(R 22 )2, wherein each R 21 is independently selected from H, and optionally substituted (C-C) alkyl, and each R 22 are independently selected from H, halogen (e.g., F), and optionally substituted (C1-C6) alkyl.

[0198] In some embodiments, Z 2 is not O.

[0199] In some embodiments of Formula (II″), Z 2 is a linking group attached in the alpha configuration to the pyranose sugar ring at the anomeric or 1-position, as shown in formula (IIa″) below: [ka]

[0200] β-linked pyranose ring The present inventors have demonstrated that although M6PR-binding compounds having an M6PR-binding moiety with the anomeric α-configuration of formula (IIa″) can provide good binding and internalization activity at the receptor, in some cases, it is possible to confer stronger binding and internalization activity at M6PR by configuring the central pyranose sugar ring of the M6PR-binding moiety with a β-configuration at the anomeric position. In some embodiments, such M6PR-binding moieties can increase the stability of the pyranose ring.

[0201] Thus, in some embodiments of formula (II″), Z 2 is a linking group attached in a β configuration to the pyranose sugar ring at the anomeric or 1-position, as shown in formula (IIb″) below: [ka]

[0202] Further M6PR ligand moieties Although the moiety of formula (II″) can exhibit binding activity to M6PR, the present inventors have discovered that certain types of cyclic groups are involved in the linking moiety Z 2 It has been demonstrated that when linked in a specific configuration adjacent to the pyranose ring of formula (II″) via

[0203] Thus, in some embodiments of formula (II″), the M6PR binding moiety (X) is of formula (III″): [ka] or a prodrug thereof, or a salt thereof, wherein: W is a hydrophilic head group; Z 1 is selected from optionally substituted (C1-C3) alkylene and optionally substituted ethenylene; Z 2 are O, S, NR 21 and C(R 22 )2, wherein each R 21 is independently selected from H, and optionally substituted (C-C) alkyl, and each R 22 are independently selected from H, halogen (e.g., F), and optionally substituted (C1-C6) alkyl; A is independently an optionally substituted cyclic group; Z 3 is independently a linking moiety.

[0204] In some embodiments of Formulas (II'')-(III''), W is a non-hydrolyzable hydrophilic head group.

[0205] In some embodiments of Formulas (II″)-(III″), Z 2 is optionally substituted ethylene. In some embodiments of Formulas (II″)-(III″), Z 2 is optionally substituted ethenylene.

[0206] In some embodiments of Formulas (II″)-(III″), Z 2 is O. In some embodiments of Formulas (II″)-(III″), Z 2 is S. In some embodiments of Formulas (II″)-(III″), Z 2 is -NR 21 In some embodiments of Formulas (II″)-(III″), Z 2 is -C(R 22 )2-, wherein each R 22 is independently selected from H, halogen (e.g., F), and optionally substituted (C1-C6) alkyl. In some embodiments of Formulas (II'')-(III''), Z 2 is -CH2-.

[0207] In some embodiments of Formulas (II")-(III"), A is an optionally substituted aryl, an optionally substituted heteroaryl, an optionally substituted heterocycle, or an optionally substituted cycloalkyl. In some embodiments of Formulas (II")-(III"), A is independently an optionally substituted aryl or heteroaryl linking moiety (e.g., an optionally substituted monocyclic or bicyclic aryl or heteroaryl).

[0208] Exemplary Z groups of formulas (II″) to (III″) 3 Linking moieties are described herein.

[0209] Such M6PR-binding moieties of formula (III") can be attached to a moiety or molecule of interest to generate bifunctional compounds that undergo efficient M6PR-mediated cellular internalization. The inventors have further demonstrated that when the moiety or molecule of interest is a target protein-binding moiety, the M6PR-binding compounds also provide M6PR-mediated internalization and / or degradation of the bound target protein.

[0210] Thus, the M6PR binding compound has the formula (XII) [ka] or a prodrug thereof, or a salt thereof, During the ceremony, W is a hydrophilic head group; Z 1 is selected from optionally substituted (C1-C3) alkylene and optionally substituted ethenylene; Z 2 are O, S, NR 21 and C(R 22 )2, wherein each R 21 is independently selected from H, and optionally substituted (C-C) alkyl, and each R 22 are independently selected from H, halogen (e.g., F), and optionally substituted (C1-C6) alkyl; A is independently an optionally substituted cyclic group; Z 3 are independently linking moieties, n is 1 to 500; L is a linker, Y is the desired part, m is 1 to 100.

[0211] In some embodiments, the cell surface M6PR binding compound has formula (XIII): [ka] or a prodrug thereof, or a salt thereof, During the ceremony, W is a hydrophilic head group; Z 1 is selected from optionally substituted (C1-C3) alkylene and optionally substituted ethenylene; Z 2 are O, S, NR 21 and C(R 22 )2, wherein each R 21 is independently selected from H, and optionally substituted (C-C) alkyl, and each R 22 are independently selected from H, halogen (e.g., F), and optionally substituted (C1-C6) alkyl; A is independently an optionally substituted cyclic group; Z 3 are independently linking moieties, n is 1 to 500; L is a linker, Y is a moiety of interest (e.g., as described herein).

[0212] In some embodiments of Formula (XIII), Y is a chemoselective ligation group. In some embodiments of Formula (XIII), n is 1. In some embodiments of Formula (XIII), Y is a chemoselective ligation group connected to "n" M6PR binding moieties (Xn-) via a single linker -L-. In some embodiments of Formula (XIII), n is 2, 3, 4, or 5. In some embodiments of Formula (XIII), n is 5 to 10. In some embodiments of Formula (XIII), n is 10 to 100, e.g., 20 to 80, 20 to 50. In some embodiments of Formula (XIII), when n is 5 or greater, L is a linker-containing polypeptide (e.g., as described herein).

[0213] In some embodiments of Formulas (XII)-(XIII), when n is 1 and A is phenyl, i) L comprises a backbone of at least 16 contiguous atoms (e.g., at least 18 contiguous atoms, or at least 20 contiguous atoms, in some cases up to about 200 contiguous atoms), ii) Y is a biomolecule, and / or ii) Z 3 is an amide, sulfonamide, urea, or thiourea linking moiety to the linker L.

[0214] In some embodiments of Formula (XII), Z 2 is a linking group attached in the alpha configuration to the sugar ring at the anomeric or 1-position as shown in formula (IIa), so that the compound has the formula (XIIa) [ka] It is of the type.

[0215] In some embodiments of formula (XII), Z is a linking group attached to the sugar ring in a β configuration at the anomeric or 1 position, as shown in formula (IIb), and the compound has formula (XIIb): [ka] It is of the type.

[0216] In some embodiments of Formula (XI)-(XIIb), multiple M6PR binding moieties, such as those of Formula (III), are linked via multiple linkers L to different ligation sites on a moiety of interest Y. In some embodiments, when Y is a biomolecule, the compounds of Formula (XI)-(XIIb) can be referred to as conjugates.

[0217] Hydrophilic Head Group and Linking Group In some embodiments of Formulas (II)-(XIII), the M6PR binding moiety (X) comprises an analog of a D-mannopyranose ring having a hydrophilic head group, or a precursor or prodrug thereof, which is linked to a linking moiety (Z 1 ) to the 5-position of the sugar ring. The length of the linking moiety may be, for example, 1 or 2 atoms, such as 1 to 6 atoms, 1 to 5, 1 to 4, or 1 to 3 atoms. It is understood that the length of the linking moiety can be selected in combination with the hydrophilic head group.

[0218] The hydrophilic head group (W) may be any suitable negatively charged group or salt thereof. In some embodiments, the hydrophilic head group is a neutral, polar, hydrophilic group. Generally, the hydrophilic head group is capable of hydrogen bonding or electrostatic interactions with M6PR under aqueous or physiological conditions, similar to the phosphate group of M6P. The hydrophilic head group may be a bioequivalent (e.g., a structural or functional mimetic) of the 6-phosphate group of a naturally occurring mannose-6-phosphate ligand. In some embodiments, the hydrophilic head group is non-hydrolyzable, i.e., under physiological conditions, the Z to which the hydrophilic head group is attached is non-hydrolyzable. 1The functional group is stable to cleavage (eg, chemical or enzymatic) from the linking moiety and / or the pyranose ring of X.

[0219] The hydrophilic head group is generally a small group such as a heteroatom-containing functional group or a single heterocycle, optionally with a MW of less than 200, eg, less than 150, or less than 100.

[0220] In some embodiments, the hydrophilic head group is a phosphonate, or a bioisostere thereof, such as a carboxylate or a malonate ester. In some embodiments, the hydrophilic head group is a thiophosphonate.

[0221] In some embodiments of Formulas (II)-(XIII), the hydrophilic head group is not a phosphate, thiophosphate, or dithiophosphate; such groups would render the compound unstable and susceptible to cleavage under physiological conditions (e.g., by phosphatases in biological systems or chemically) due to the phosphate ester linkage. For example, the 6-phosphate group of M6P exhibits undesirable stability compared to phosphonate analogs or other more stable head groups. The present disclosure also provides alternative non-hydrolyzable head groups in addition to phosphonates that retain the binding and internalization activity of the resulting M6PR-binding compounds.

[0222] In any one of the embodiments of formulas (II)-(XIII), the hydrophilic head group W is selected from the group consisting of -OH, -CR 2 R 2 OH, -NR 3 P=O(OH)2, -P=O(OH) 2、 -P=S(OH) 2、 -P=O(SH)(OH), -P=S(SH)(OH), P(=O)R 1 OH, -PH(=O)OH, -(CR 2 R 2 )-P=O(OH)2, -SO2OH (i.e., -SO3H), -S(O)OH, -OSO2OH, -COOH, -CN, -CONH2, -CONHR 3 , -CONR 3 R4 , -CONH(OH), -CONH(OR 3 ), -CONHSO2R 3、 -CONHSO2NR 3 R 4 , -CH(COOH)2, -CR 1 R 2 COOH, -SO2R 3 , -SOR 3 R 4 , -SO2NH 2、 -SO2NHR 3 , -SO2NR 3 R 4 , -SO2NHCOR 3 , -NHCOR 3 , -NHC(O)CO2H, -NHSO2NHR 3 , -NHC(O)NHS(O)2R 3 , -NHSO2R 3 , -NHSO3H, [ka] or salts thereof, wherein: R 1 and R 2 are independently hydrogen, SR 3 , halo, or CN, R 3 and R 4 are independently H, C 1~6 Alkyl or substituted C 1~6 alkyl (e.g., —CF or —CHCF), A, B, and C are each independently CH or N; Each D is independently O or S.

[0223] In some embodiments of Formulas (II)-(XIII), the hydrophilic head group W is a phosphate or thiophosphate group, e.g., -OP=O(OH), -SP=O(OH), -OP=O(SH)(OH), -SP=O(SH)(OH), -OP=S(OH), -OP=O(N(R 3 )2)(OH), or -OP=O(R 3)(OH), or a salt thereof. In some embodiments of Formulas (II)-(XIII), the hydrophilic head group W is non-hydrolyzable and therefore is a phosphate or thiophosphate, e.g., -OP=O(OH), -SP=O(OH), -OP=O(SH)(OH), -SP=O(SH)(OH), -OP=S(OH), -OP=O(N(R 3 )2)(OH), or -OP=O(R 3 )(OH), or salts thereof.

[0224] In some embodiments of Formulas (II)-(XIII), the hydrophilic head group W is charged and capable of forming salts, e.g., under aqueous or physiological conditions. In some embodiments of Formulas (II)-(XIII), the hydrophilic head group W is -NR 3 P=O(OH)2, -P=O(OH)2, -P=S(OH)2, -P=O(SH)(OH), -P=S(SH)(OH), P(=O)R 1 OH, -PH(=O)OH, -(CR 2 R 2 )-P=O(OH)2, -COOH, -CH(COOH)2, -CR 1 R 2 COOH, and —NHC(O)COH.

[0225] In some embodiments of Formulas (II)-(XIII), the hydrophilic head group W is a phosphonate or thiophosphonate (e.g., -P=O(OH)2, -P=S(OH)2, -P=O(SH)(OH), or -P=S(SH)(OH), or a salt thereof). In some embodiments of Formulas (II)-(XIII), the hydrophilic head group W is a phosphonate or a salt thereof. In some embodiments of Formulas (II)-(XIII), the hydrophilic head group W is -CO2H or a salt thereof. In some embodiments of Formulas (II)-(XIII), the hydrophilic head group W is a malonate ester (e.g., -CH(COOH)2 or a salt thereof).

[0226] In some embodiments of Formulas (II)-(XIII), the hydrophilic head group W is selected from -SO2OH (i.e., -SO3H), -S(O)OH, -OSO2OH, and -NHSO3H. In some embodiments of Formulas (II)-(XIII), the hydrophilic head group W is a sulfonate (e.g., -SO3H or a salt thereof).

[0227] In some embodiments, the hydrophilic head group W is neutrally hydrophilic. In some embodiments of Formulas (II)-(XIII), the hydrophilic head group W is -OH, -CR 2 R 2 OH, -CN, -CONH2, -CONHR 3 , -CONR 3 R 4 , -CONH(OH), -CONH(OR 3 ), -CONHSO2R 3 , -SO2R 3 , -SOR 3 R 4 , -SO2NH2, -SO2NHR3, -SO2NR 3 R 4 , -SO2NHCOR 3 , -NHCOR 3 , -NHSO2NHR 3 , -NHC(O)NHS(O)2R 3 , and -NHSO2R 3 is selected from.

[0228] In some embodiments of Formulas (II)-(XIII), the hydrophilic head group W is a heterocycle, e.g., [ka] or salts thereof, wherein: A, B, and C are each independently CH or N, and D is each independently O or S.

[0229] In some embodiments of Formulas (II)-(XIII), the hydrophilic head group W is a five-membered heterocycle, such as [ka] or salts thereof.

[0230] In some embodiments of Formulas (II)-(XIII), the hydrophilic head group W is selected from optionally substituted (C1-C2) alkylene and optionally substituted ethenylene. 1 It is connected to the pyranose ring via Z. 1 Z can be selected in combination with W to provide the desired spacing between the 5-position of the ring and the charged or polar center of W. For example, if W is a malonate having a C-H atom connecting the two carboxylic acid groups, then Z 1 may be methylene, which together provide the desired two-carbon spacer between the ring and the COOH group.

[0231] In some embodiments of Formulas (II)-(XIII), Z 1 is methylene or substituted methylene. In some embodiments of Formulas (II)-(XIII), Z 1 is ethyl or substituted ethyl. In some embodiments of Formulas (II)-(XIII), Z 1 is ethenylene or substituted ethenylene. In some embodiments of Formulas (II)-(XIII), Z 1 is substituted with one or more halogens, for example fluorine.

[0232] In some embodiments of formula (III), the M6PR binding moiety (X) is selected from the group consisting of formulas (IV-1) to (IV-3): [ka] wherein R a , R b , R c and R d are independently H or F.

[0233] In some embodiments of formulas (IV-1) to (IV-3), Z 2 is O.

[0234] In some embodiments of formulas (IV-1) to (IV-3), Z 2 is S.

[0235] In some embodiments of formulas (IV)-1 to (IV-3), Z 2 is -NR 21 -It is.

[0236] In some embodiments of formulas (IV-1) to (IV-3), Z 2 is -C(R 22 )2-, wherein each R 22 is independently selected from H, halogen (e.g., F), and optionally substituted (C1-C6) alkyl. In some embodiments of formulas (IV-1) to (IV-3), Z 2 is -CH2-.

[0237] In some embodiments of formulas (IV-1) to (IV-3), R a , R b , R c and R d are H respectively.

[0238] In some embodiments of formula (IV-1), R a is H, R b is F. In some embodiments of formula (IV-1), R a and R b are F respectively.

[0239] In some embodiments of formula (IV-2), R c is H. In some embodiments of formula (IV-2), R c is F.

[0240] In some embodiments of formula (IV-3), R d is H. In some embodiments of formula (IV-3), R d is F.

[0241] In some embodiments of Formula (IV-1) and (IV-3), W is selected from -P=O(OH)2, -P=S(OH)2, -P=O(SH)(OH), -P=S(SH)(OH), and -COOH, or a salt thereof. In some embodiments of Formula (IV-1) and (IV-3), W is -P=O(OH)2, or a salt thereof. In some embodiments of Formula (IV-1) and (IV-3), W is COOH, or a salt thereof.

[0242] In some embodiments of formula (IV-1), R a and R b are each F, and W is -P=O(OH)2, or a salt thereof. In some embodiments of formula (IV-1), R a and R b are each H, and W is -P=O(OH)2, or a salt thereof. In some embodiments of formula (IV-1), R a is F and R b is H and W is -P=O(OH)2, or a salt thereof.

[0243] In some embodiments of formulas (IV-1) to (IV-3), Z 2 is linked to the anomeric position of the pyranose ring in the α-configuration. In such cases, the M6PR binding moieties (X) of (IV-1) to (IV-3) are represented by formulas (IV-A1) to (IV-A3), respectively.

[0244] In some embodiments of formulas (IV-A1) to (IV-A3), Z 2 is S. In some embodiments of formulas (IV-A1) to (IV-A3), Z 2 is O. In some embodiments of formulas (IV-A1) to (IV-A3), Z 2 In some embodiments of formulas (IV-A1) to (IV-A3), Z 2 is -CF2-.

[0245] In some embodiments of Formula (IV-A1) and (IV-A3), W is selected from -P=O(OH)2, -P=S(OH)2, -P=O(SH)(OH), -P=S(SH)(OH), and -COOH, or a salt thereof. In some embodiments of Formula (IV-A1) and (IV-A3), W is -P=O(OH)2, or a salt thereof. In some embodiments of Formula (IV-A1) and (IV-A3), W is COOH, or a salt thereof.

[0246] In some embodiments of formula (IV-A1), R a and R b are each F and W is -P=O(OH)2, or a salt thereof. In some embodiments of formula (IV-A1), R a and R b are each H and W is -P=O(OH)2, or a salt thereof. In some embodiments of formula (IV-A1), R a is F and R b is H and W is -P=O(OH)2, or a salt thereof.

[0247] In some embodiments of formulas (IV-1) to (IV-3), Z 2 is linked in a β-configuration to the anomeric position of the pyranose ring. The inventors have demonstrated that compounds comprising M6PR-binding moieties with a β-glycosidic configuration can have at least equivalent binding activity and / or cellular uptake activity compared to conjugates with corresponding α-glycosidic configurations. In some embodiments, such M6PR-binding moieties with a β-glycosidic configuration can have improved stability compared to reference compounds with a β-glycosidic configuration. Thus, in some embodiments of formula (IV), the M6PR-binding moiety (X) is represented by formulas (IV-B1) to (IV-B3): [ka] wherein R a , R b , R c and R dare independently H or F.

[0248] In some embodiments of formulas (IV-B1) to (IV-B3), Z 2 is S. In some embodiments of formulas (IV-B1) to (IV-B3), Z 2 is O. In some embodiments of formulas (IV-B1) to (IV-B3), Z 2 In some embodiments of formulas (IV-B1) to (IV-B3), Z 2 is -CF2-.

[0249] In some embodiments of Formula (IV-B1) and (IV-B3), W is selected from -P=O(OH)2, -P=S(OH)2, -P=O(SH)(OH), -P=S(SH)(OH), and -COOH, or a salt thereof.

[0250] In some embodiments of Formula (IV-B1) and (IV-B3), W is selected from -P=O(OH)2, -P=S(OH)2, -P=O(SH)(OH), -P=S(SH)(OH), and -COOH, or a salt thereof. In some embodiments of Formula (IV-B1) and (IV-B3), W is -P=O(OH)2, or a salt thereof. In some embodiments of Formula (IV-B1) and (IV-B3), W is COOH, or a salt thereof.

[0251] In some embodiments of formula (IV-B1), R a and R b are each F and W is -P=O(OH)2, or a salt thereof. In some embodiments of Formula (IV-B1), R a and R b are each H and W is -P=O(OH)2, or a salt thereof. In some embodiments of Formula (IV-B1), R a is F and R b is H and W is -P=O(OH)2, or a salt thereof.

[0252] The present inventors have demonstrated that conjugates comprising M6PR-binding moieties with a β-S-glycoside configuration can have at least equivalent or superior binding and / or cellular uptake activity compared to conjugates with the corresponding α-S-glycoside configuration or α-O-glycoside configuration.

[0253] Thus, in some embodiments of formulas (IV-B1) to (IV-B3), the M6PR binding moiety (X) is represented by formula (IV-BS1) to (IV-BS3): [ka] wherein R a , R b , R c and R d are independently H or F.

[0254] In some embodiments of formulas (IV-BS1) to (IV-BS3), R a , R b , R c and R d are H respectively.

[0255] In some embodiments of Formula (IV-BS1), R a is H, R b is F. In some embodiments of formula (IV-BS1), R a and R b are F respectively.

[0256] In some embodiments of Formula (IV-BS2), R c is H. In some embodiments of formula (IV-B2), R c is F.

[0257] In some embodiments of Formula (IV-BS3), R d is H. In some embodiments of formula (IV-BS3), R d is F.

[0258] In some embodiments of formulas (IV-BS1) to (IV-BS3), Z 2 is S. In some embodiments of formulas (IV-BS1) to (IV-BS3), Z 2 is O. In some embodiments of formulas (IV-BS1) to (IV-BS3), Z 2 In some embodiments of formulas (IV-BS1) to (IV-BS3), Z 2 is -CF2-.

[0259] In some embodiments of Formula (IV-BS1) and (IV-BS3), W is selected from -P=O(OH)2, -P=S(OH)2, -P=O(SH)(OH), -P=S(SH)(OH), and -COOH, or a salt thereof. In some embodiments of Formula (IV-BS1) and (IV-BS3), W is -P=O(OH)2, or a salt thereof. In some embodiments of Formula (IV-BS1) and (IV-BS3), W is COOH, or a salt thereof.

[0260] In some embodiments of Formula (IV-BS1), R a and R b are each F and W is -P=O(OH)2, or a salt thereof. In some embodiments of Formula (IV-BS1), R a and R b are each H and W is -P=O(OH)2, or a salt thereof. In some embodiments of Formula (IV-BS1), R a is F and R b is H and W is -P=O(OH)2, or a salt thereof.

[0261] In some embodiments, the mannose ring or analogue thereof of the M6PR binding moiety has a Z attached at the anomeric or 1 position of the sugar ring. 2 Linking moieties can be attached to groups to be incorporated into compounds of the present disclosure.

[0262] In some embodiments, the M6PR binding moiety is a Z attached to a cyclic group A. 3In the compounds of formula (III), Z is incorporated into the compounds of the present disclosure by attaching a linker to the group. 2 The cyclic group attached to is considered part of the M6PR binding moiety (X) and is believed to provide the desired binding properties to M6PR.

[0263] Cyclic Group A The A cyclic group of formulas (III)-(XIII) may be a monocyclic group or a bicyclic group. The bicyclic group of interest may be a fused bicyclic group or a bicyclic group containing two monocyclic groups linked by a covalent bond. The A cyclic group of formulas (III)-(XIII) may be an optionally substituted aryl, an optionally substituted heteroaryl, an optionally substituted heterocycle (e.g., a saturated heterocycle), or an optionally substituted cycloalkyl.

[0264] The A cyclic group in Formulas (III)-(XIII) may be a monocyclic aryl group or a monocyclic heteroaryl group. In some embodiments of Formulas (III)-(XIII), A is a 5-membered monocyclic heteroaryl group. In some embodiments of Formulas (III)-(XIII), A is a 6-membered monocyclic aryl or heteroaryl group. In some embodiments of Formulas (III)-(XIII), A may be a polycyclic aryl or polycyclic heteroaryl group, such as a bicyclic aryl or bicyclic heteroaryl group. In some embodiments of Formulas (III)-(XIII), A is a fused bicyclic group. In some embodiments of Formulas (III)-(XIII), A is a bicyclic group comprising two aryl and / or heteroaryl monocyclic rings connected by a covalent bond. In some embodiments of Formulas (III)-(XIII), A is a bicyclic aryl or bicyclic heteroaryl group having two 6-membered rings. In some embodiments of Formulas (III)-(XIII), A is a bicyclic aryl or bicyclic heteroaryl group having one 6-membered ring covalently attached or fused to a 5-membered ring.

[0265] In some embodiments of Formulas (III)-(XIII), A is selected from optionally substituted phenyl, optionally substituted pyridyl, optionally substituted biphenyl, optionally substituted naphthalene, optionally substituted quinoline, optionally substituted triazole, and optionally substituted phenylene-triazole.

[0266] In some embodiments of Formula (III)-(XIII), A is not phenyl (also called phenylene in the context of Formula (III), eg, 1,4-phenylene).

[0267] In some embodiments of Formulas (III)-(XIII), A is substituted with at least one OH substituent. In some embodiments of Formulas (III)-(XIII), A is substituted with one, two, or more OH groups. In some embodiments of Formulas (III)-(XIII), A is substituted with at least one optionally substituted (C1-C6) alkyl.

[0268] In some embodiments of Formulas (III)-(XIII), A is optionally substituted 1,4-phenylene, optionally substituted 1,3-phenylene, or optionally substituted 2,5-pyridylene.

[0269] In some embodiments of Formulas (III)-(XIII), A is [ka] is selected from: R 11 ~R 14 is H, halogen, OH, optionally substituted (C1-C6) alkyl, optionally substituted (C1-C6) alkoxy, COOH, NO2, CN, NH2, -N(R 25 )2, -OCOR 25 , -COOR 25 , -CONHR 25 , and -NHCOR 25 are independently selected from R 25are independently selected from H, and optionally substituted (C1-C6) alkyl.

[0270] In some embodiments of Formulas (III)-(XIII), A is an optionally substituted fused bicyclic aryl or an optionally substituted fused bicyclic heteroaryl.

[0271] In some embodiments of Formulas (III)-(XIII), A is an optionally substituted naphthalene or an optionally substituted quinoline.

[0272] In some embodiments of Formulas (III)-(XIII), A is [ka] is selected from: R 11 and R 13 ~R 14 is H, halogen, OH, optionally substituted (C1-C6) alkyl, optionally substituted (C1-C6) alkoxy, COOH, NO2, CN, NH2, -N(R 25 )2, -OCOR 25 , -COOR 25 , -CONHR 25 , and -NHCOR 25 are independently selected from s is 0 to 3, Each R 25 are independently selected from H, and optionally substituted (C1-C6) alkyl.

[0273] In some embodiments of Formulas (III)-(XIII), A is [ka] is selected from.

[0274] In some embodiments of formulas (III)-(XIII), A is an optionally substituted bicyclic aryl or an optionally substituted bicyclic heteroaryl of the following formula: [ka] or a salt thereof, wherein: Cy is independently a monocyclic aryl or a monocyclic heteroaryl; R 11 ~R 15 is H, halogen, OH, optionally substituted (C1-C6) alkyl, optionally substituted (C1-C6) alkoxy, COOH, NO2, CN, NH2, -N(R 25 )2, -OCOR 25 , -COOR 25 , -CONHR 25 , and -NHCOR 25 are independently selected from s is 0 to 4, Each R 25 are independently selected from H, and optionally substituted (C1-C6) alkyl.

[0275] In some embodiments, when Cy is optionally substituted phenyl, A is a group of formula [ka] is an optionally substituted biphenyl of the formula:

[0276] In some embodiments of Formulas (III)-(XIII), A is [ka] is selected from.

[0277] In some embodiments, when Cy is triazole, A is [ka] is selected from.

[0278] In some embodiments, R 11 ~R 15 At least one of is OH (eg, at least two are OH).

[0279] In some embodiments, R 11 ~R 15 are H, respectively.

[0280] Connecting part Z 3 Connecting part Z 3 may be any convenient linking moiety that connects the linker L to the cyclic ring A. In some embodiments of Formulas (III)-(XIII), Z 3 has a backbone of three or fewer atoms.

[0281] In some embodiments of Formulas (III)-(XIII), Z 3 is a covalent bond, -O-, -NR 23 -, -NR 23 CO-, -CONR 23 -, -NR 23 CO2-, -OCONR 23 , -NR 23 C(=X 1 )NR 23 -, -CR 24 =N-, -CR 24 =NX 2 , -N(R 23 )SO2- and -SO2N(R 23 )-, wherein X 1 and X 2 are O, S and NR 23 Selected from R 23 and R 24 is H, C (1~3) -Alkyl (e.g., methyl) and substituted C (1~3) -alkyl.

[0282] In some embodiments of Formulas (III)-(XIII), Z 3 is the covalent bond connecting A to L.

[0283] In some embodiments of Formulas (III)-(XIII), Z 3 is an optionally substituted amide, urea or thiourea.

[0284] In some embodiments of Formulas (III)-(XIII), Z 3 teeth, [ka] where: X 1 is O or S, t is 0 or 1, Each R 23 is H, C (1~3) -Alkyl (e.g., methyl or ethyl) and substituted C (1~3) -alkyl. 3 In some embodiments, X 1 is O. Z 3 In some embodiments, X 1 is S. Z 3 In some embodiments, t is 0, X 1 is O, and as a result, Z 3 is an amide. 3 In some embodiments, t is 1, such that Z 3 is urea or thiourea.

[0285] In some embodiments of Formulas (III)-(XIII), Z 3 is -N(R 23 )SO2- or -SO2N(R 23 In some embodiments of Formulas (III)-(XIII), Z 3 is -NHSO2- or -SO2NH-.

[0286] In some embodiments of Formulas (III)-(XIII), Z 3 is -N(R 23 )CO- or -CON(R 23 In some embodiments of Formulas (III)-(XIII), Z 3 is -NHCO- or -CONH-.

[0287] In some embodiments of Formulas (III)-(XIII), Z 3 is -NHC(=X 1 )NH—, wherein X 1 is O or S. In some embodiments, X 1 is O (i.e., Z 3 is —NHC(═O)NH—). In some embodiments, X 1 is S.

[0288] In some embodiments of Formulas (III)-(XIII), Z 3 is an optionally substituted triazole. 3 When is an optionally substituted triazole, it can be synthetically derived from click chemistry coupling of an azide-containing precursor with an alkyne-containing precursor of the compound.

[0289] In some embodiments, Z 3 represents the cyclic group A and / or the linking moiety Z 1 are selected in combination with each other to confer desirable M6PR binding and internalization properties on X.

[0290] In some embodiments of Formulas (III)-(XIII), -AZ 3 -teeth, [ka] is selected from.

[0291] In some embodiments of Formulas (III)-(XIII), -AZ 3 -teeth, [ka] is selected from.

[0292] In some embodiments of Formulas (III)-(XIII), -AZ 3 -teeth, [ka] is selected from.

[0293] In some embodiments of Formulas (II)-(XIb), -AZ 3 -teeth, [ka] is selected from.

[0294] In some embodiments of Formulas (III)-(XIII), -AZ 3 -teeth, [ka] [ka] is selected from.

[0295] In some embodiments of Formulas (III)-(XIII), Z 2 is O.

[0296] In some embodiments of Formulas (III)-(XIII), Z 2 is S.

[0297] In some embodiments of Formulas (III)-(XIII), Z 2 is -NR 21 -It is.

[0298] In some embodiments of Formulas (III)-(XIII), Z 2 is -C(R 22 )2-, wherein each R 22 is independently selected from H, halogen (e.g., F), and optionally substituted (C1-C6) alkyl. In some embodiments, Z 2 is -CH-. In some embodiments, Z 2 is CHF-. In some embodiments, Z 2 is -CF2-.

[0299] In some embodiments of Formulas (III)-(XIII), Z 2 -A-Z 3 - is as follows: [ka] During the ceremony, Z 21 - is O, S, or -C(R 22 )2- and R 16 is OH or CH3, w is 0 to 4 (for example, w is 0, 1, or 2).

[0300] In some embodiments, Z 21 is S or O. In some embodiments, Z 21 is -CH-. In some embodiments, Z 21 is -CHF-. In some embodiments, Z 21 is -CF-. In some embodiments, R 16 is OH and w is 1. In some embodiments, R 16 is CH3 and w is 1. In some embodiments, w is 0.

[0301] In some embodiments of Formulas (III)-(XIII), -Z 2 -A-Z 3 -teeth, [ka] is.

[0302] In some embodiments of Formulas (III)-(XII), -Z 2 -A-Z 3 -teeth, [ka] is.

[0303] In some embodiments of Formulas (III)-(XIII), -Z 2 -A-Z3 -teeth, [ka] is.

[0304] In some embodiments of Formulas (III)-(XIII), -Z 2 -A-Z 3 -teeth, [ka] is.

[0305] In some embodiments of Formulas (III)-(XIII), -Z 2 -A-Z 3 -teeth, [ka] is.

[0306] In some embodiments of Formulas (III)-(XIII), -Z 2 -A-Z 3 -teeth, [ka] is.

[0307] Prodrug Aspects of the present disclosure include any prodrugs of the ASGPR and M6PR binding moieties described herein that are incorporated into the compounds and conjugates of the present disclosure.

[0308] The term "prodrug" refers to a substance that is converted into a drug within the body by some physiological or chemical process (e.g., a prodrug is converted into the desired drug form when adjusted to physiological pH).

[0309] Prodrug forms of any of the ASGPR or M6PR binding moieties described herein can be useful because they can provide 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 required active dose.

[0310] Prodrugs may be useful in some situations because they may be easier to administer than the parent drug. For example, they may be oral bioavailable whereas the parent drug is not. The prodrug may also have improved solubility in pharmacological compositions over the parent drug.

[0311] Prodrug derivatives of ASGPR or M6PR binding moieties generally contain a promoiety substituent at an appropriate labile site on the compound, where the promoiety refers to a group that is removed by enzymatic or chemical reaction when the prodrug is converted to a drug in the body.

[0312] In some embodiments, the promoiety is a group attached to a hydroxyl group of a compound or drug via an ester linkage.

[0313] In some embodiments, prodrug derivatives of one or more hydroxyl groups of the sugar ring can be incorporated into the compound. For example, an ester promoiety can be incorporated into one or more of the 3- and / or 4-hydroxyl groups of the sugar (e.g., as described herein). In some embodiments, the 3- and 4-hydroxyl groups of the sugar are cyclically linked to form a promoiety (e.g., as described herein).

[0314] The terms "linker," "linking moiety," and "linking group" are used interchangeably and refer to a linking moiety that covalently bonds two or more moieties, compounds, or other biomolecules, such as a ligand and a protein of interest. In some cases, the linker is divalent and connects two moieties. In certain cases, the linker is a trivalent or higher polyvalent, branched linking group. In some cases, the linker connecting two or more moieties has a linear or branched backbone with a length measured between the two or more moieties of 500 atoms or less (e.g., 400 atoms or less, 300 atoms or less, 200 atoms or less, 100 atoms or less, 80 atoms or less, 60 atoms or less, 50 atoms or less, 40 atoms or less, 30 atoms or less, or 20 atoms or less). The linking moiety can be a covalent bond connecting two groups, or a linear or branched chain between 1 and 500 atoms in length, e.g., a chain of 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 linear, branched, cyclic, or single-atom. In certain cases, 1, 2, 3, 4, 5, or more, 10, or even more carbon atoms in the linker backbone can be optionally substituted with heteroatoms, such as sulfur, nitrogen, or oxygen heteroatoms. In certain situations, the linker includes an ethylene glycol or longer polyethylene glycol (PEG) linking group, for example, when every third atom in that segment of the linker backbone is substituted with oxygen. The bonds between the backbone atoms of the linker can be saturated or unsaturated, and typically there are no more than one, two, or three unsaturated bonds in the linker backbone. Linkers can include one or more substituents, such as, for example, alkyl groups, aryl groups, alkenyl groups, etc. Linkers include, but are not limited to, one or more of oligo(ethylene glycol) (also known as PEG), ether, thioether, disulfide, amide, carbonate, carbamate, urea, sulfonamide, thiourea, tertiary amine, linear or branched alkyl (e.g., methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl), etc.).The linker backbone comprises a cyclic group, such as an aryl, heterocycle, cycloalkyl group or hetero group, and more than one atom of the cyclic group, such as 2, 3 or 4 atoms, is included in the backbone.

[0315] In some embodiments, the "linker" or linking moiety is derived from a molecule having a reactive terminus suitable for conjugation to, for example, a protein of interest. In certain circumstances, the reactive terminus of the linker precursor comprises a chemoselective ligation group capable of conjugation to an amino acid residue(s) of a polypeptide. In certain circumstances, the chemoselective ligation group is conjugated to an accessible cysteine ​​thiol group or lysine side chain amine group of a polypeptide. A variety of conjugation chemistries can be utilized in the conjugates of the present disclosure (e.g., as described herein). In some embodiments, the chemoselective ligation group is a thiol-reactive group such as maleimide or dibromomaleimide. In some embodiments, the chemoselective ligation group is an active ester, such as a perfluorophenyl ester or tetrafluorophenyl ester, or N-hydroxysuccinimidyl ester (NHS), or sulfo-NHS, or an amine-reactive group as defined herein.

[0316] 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—), as well as combinations thereof. In certain embodiments, these linkers optionally have an amide bond, a urea or thiourea linkage, a carbamate linkage, an ester linkage, an amino linkage, an ether linkage, a thioether linkage, a sulfhydryl linkage, a heteroaryl linkage, or other heterofunctional linkage. In certain embodiments, the linker backbone 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. In certain cases, the linker includes one or more heteroaryl ring structures, for example, a triazole, such as a 1,2,3-triazole.

[0317] In certain embodiments, L is a linker between about 5 Å and about 500 Å. In certain embodiments, L is between about 10 Å and about 400 Å. In certain embodiments, L is between about 10 Å and about 300 Å. In certain embodiments, L is between about 10 Å and about 200 Å. In certain embodiments, L is between about 10 Å and about 100 Å.

[0318] In certain embodiments, the linker L is connected to X (or Z) by a chain of 10 to 100 contiguous atoms. 1 ) and Y. In certain embodiments, the linker L separates X (or Z 1) and Y are separated by a chain of 10 to 60 consecutive atoms, a chain of 12 to 60 consecutive atoms, a chain of 16 to 50 consecutive atoms, a chain of 20 to 50 consecutive atoms, a chain of 30 to 50 consecutive atoms, or a chain of 40 to 50 consecutive atoms.

[0319] The linker is Z of the ASGPR ligand portion (X). 1 It is understood that the linker may be considered to be directly connected to the Z group (e.g., as described herein). In some embodiments of Formula II (or any formula described herein for the ASGPR ligand moiety (X)), the linker is 1 Alternatively, the -Z 1 -L 1 The - group (e.g., as described herein) can be considered part of the linking moiety connecting L to Y. The present disclosure is meant to include all such arrangements of ASGPR ligand moieties (X) and linkers (L).

[0320] In some embodiments of formula (I), L is a group represented by formula (LXI): [ka] is a linker of the formula: L 1 and L 3 are each independently a linear linking moiety, and L 2 is a branched linking moiety, and in the formula, 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 Z 1 X's L via 1 represents the attachment point to ** represents the conjugation point between the linker L and Y; During the ceremony, When n is 1, b is 0 and at least one of a and c is 1; When n is 2 or 3, a, b, and c are each 1.

[0321] In some embodiments of the linker of formula (LXI), n is 1, a is 1, b is 0, and c is 1, such that the linker L has the formula (LXia): [ka] It is of the type.

[0322] In some embodiments of a linker of formula (LXI), n is 1, a is 1, b is 0, and c is 1, such that the linker L has the formula [ka] It is of the type.

[0323] In certain embodiments, the linear linker of formula (LXia) comprises a Z backbone, such as a backbone of 12 or more consecutive atoms, 14 or more consecutive atoms, or 16 or more consecutive atoms, in some cases up to 100 consecutive atoms. 1 In certain embodiments of Formula (LXia), the linear linker has a backbone of 10 or more consecutive atoms covalently linking X to Y via a chain of 20 to 50 consecutive atoms. 1 ) and Y. In certain embodiments of Formula (LXa), the linear linker L separates X (or Z) from Y by a chain of 30 to 60 contiguous atoms. 1 ) and Y.

[0324] In some embodiments of the linker of formula (LXI), n is 2, a is 1, b is 1, and c is 1, such that the linker L has the formula (LXib): [ka] It is of the type.

[0325] In some embodiments of the linker of formula (LXI), n is 3, a is 1, b is 1, and c is 1, such that the linker L has the formula (LXic): [ka] It is of the type.

[0326] In some embodiments of a linker of any one of formulas (LXI) or (LXia) through (LXic), each L 1 is represented by formula (LXII) [ka] wherein: L 10 is the linking part, * is Z 1 X's L via 1 represents the attachment point to L 11 ~L 19 is independently absent or a linking moiety, In the formula, each L 1 L 10 ~L 19 -C 1~6 -Alkylene-, -CF2-, -C 1~12 -Alkylene-, -C 1~20 -Alkylene-, -NHCO-C 1~6 -Alkylene-, -CONH-C 1~6 -Alkylene-, -NH-C 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 -, -NHCO-, -CONH-, -NHSO2-, -SO2NH-, -NHCONH-, -NHCSNH-, -CO-, -SO2-, -O-, -S-, pyrrolidine-2,5-dione, 1,2,3-triazole, -NH-, -N(C 1~6-alkyl)-, and -N(CH3)-, where each p is independently 1 to 50, e.g., 1 to 20, 1 to 12, 1 to 10, 1 to 8, or 1 to 6, e.g., 1, 2, 3, 4, 5, or 6.

[0327] In certain embodiments of Formula (LXII), the linking moiety L 1 comprises a linear backbone of 6 to 40 consecutive atoms, e.g., 10 to 40, 10 to 30, 16 to 30, or 20 to 30 consecutive atoms. In certain embodiments of Formula (LXII), the linking moiety L 1 comprises a linear backbone of 6 to 20 consecutive atoms, for example 6 to 16 consecutive atoms, for example 8, 9, 10, 11, 12, 13, 14, 15, or 16 consecutive atoms.

[0328] In certain embodiments, the linking moiety of formula (LXII) comprises one or more repeating ethylene glycol moieties (e.g., -CH2CHO- or -OCH2CH2-). In certain cases, the linking moiety of formula (XII) comprises 1 to 10 ethylene glycol moieties, e.g., 1, 2, 3, 4, 5, or 6 ethylene glycol moieties.

[0329] In certain embodiments, the linking moiety of formula (LXII) comprises one or more triazole (e.g., 1,2,3-triazole) containing linking moieties. It is understood that the triazoles can be derived from azide-alkyne click chemistry and thus have two possible directions depending on the synthetic method: [ka]

[0330] In certain embodiments, the triazole-containing linking moiety is: [ka] where w1 and u1 are independently a value from 0 to 12, such as 0, 1, 2, 3, 4, 5, or 6.

[0331] In some embodiments of the linker of formula (LXI), b is 1 and L 2 is represented by formula (LXIIIa) or (LXIIIb) [ka] wherein: L 20 is an amino acid residue (e.g., residues such as Gly, Ala, beta-Ala, Lys, Orn, Asp, Glu, Ser, Cys, or derivatives thereof), -NH-CH[(CH2) q ]2O- or -NH-C[(CH2) q ]3O-, [ka] -C 1~6 -Alkylene-, -NHCO-, -CONH-, -NHSO2-, -SO2NH-, -CO-, -SO2-, -O-, -S-, pyrrolidine-2,5-dione, 1,2,3-triazole, -NH-, and -Nme-, -NHC(O)NH-, -NHC(S)NH-, -O(CH2) p - and -(OCH2CH2) p - a branched linking moiety comprising one or more linking moieties independently selected from In the formula, each p is independently 1 to 50, and q is 1 to 6.

[0332] In some embodiments of the linker of formula (LXI), b is 1 and the linking moiety L 2 is selected from one of (L2A) to (L2D), [ka] During the ceremony, Z 2 and Z 3 are each independently selected from absent or -NHCO-, -CONH-, -CO-, -O-, -NH-, and -Nme-; x is 1 to 12 (e.g., 1 to 6, or 1 to 3), y is 0 to 12 (for example, 1 to 6, or 1 to 3).

[0333] In some embodiments of any one of L2A to L2D, Z 2 In some embodiments of any one of L2A-L2D, Z 2 In some embodiments of any one of L2A to L2D, Z 2 In some embodiments of any one of L2A to L2D, Z 2 is —O—. In some embodiments of any one of L2A-L2D, Z 2 In some embodiments of any one of L2A-L2D, Z 2 In some embodiments of any one of L2A to L2D, Z 2 does not exist.

[0334] In some embodiments of any one of L2A to L2D, Z 3 In some embodiments of any one of L2A-L2D, Z 3 In some embodiments of any one of L2A to L2D, Z 3 In some embodiments of any one of L2A to L2D, Z 3 is —O—. In some embodiments of any one of L2A-L2D, Z 3 In some embodiments of any one of L2A-L2D, Z 3 In some embodiments of any one of L2A to L2D, Z 3 does not exist.

[0335] In some embodiments of L2A, Z 2 is -O-, y is 0, and the linking moiety is of the structure L2Ai [ka] It is of the type.

[0336] In some embodiments of L2B, Z 2 is -O- or -CO-, and the linking moiety is structure L2Bi or L2Bii [ka] It is of the type.

[0337] In some embodiments of L2C, Z 2 is -O-, -CO-, -NHCO-, or -NH-, and the linking moiety is of structure L2Ci, L2Cii, L2Ciii, or L2Civ [ka] It is of the type.

[0338] In some embodiments of L2D, Z 2 does not exist, and the linking part has the structure L2Di [ka] It is of the type.

[0339] In some embodiments of any one of formulas L2A through L2Di, x is 1 to 6. Optionally, x is 1 to 3. Optionally, x is 1. Optionally, x is 2. Optionally, x is 3.

[0340] In some embodiments of any one of Formulas L2A through L2Di, y is 0 to 6. In some cases, y is 0 to 3. In some cases, y is 0. In some cases, y is 1. In some cases, y is 2. In some cases, y is 3.

[0341] In some embodiments of Formula (LXI), b is 1 and the linking moiety L 2 teeth, [ka] is selected from.

[0342] In some embodiments of the linker of formula (LXI), b is 1 and the linking moiety L 2 is represented by formula (LXIV) [ka] wherein: r is 1 or 2, If n is 2, then r is 1; If n is 3, then r is 2.

[0343] In some embodiments of the linker of formula (LXI), b is 1 and the linking moiety L 2 is the formula (LXva) or (LXVb) [ka] wherein: r is 1 or 2, If n is 2, then r is 1; If n is 3, then r is 2.

[0344] In some embodiments, L 2 is of formula (LXIIIa) or (LXIIIb), and L 2 comprises two or more amino acid residues (e.g., three or more, or four or more amino acid residues, linear or dendrimer). 2 comprises four or more amino acid residues that are branched linking moieties selected from Lys, Orn, Asp, Glu, Ser, and Cys (e.g., where the side chain, amino, and carboxylic acid are each linked to adjacent moieties).

[0345] In some embodiments of the linker of any one of Formulas (LXI) or (LXa)-(LXc), each L 3 is represented by formula (LXVI) [ka] wherein: L 30 ~L 39 is independently absent or a linking moiety, Z is a residue moiety resulting from the covalent bond between the chemoselective ligation group of the linker and the compatible group of Y; In the formula, L 30 ~L 39 are -C respectively. 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 -,-(OCH2CH 2 ) p and independently selected from -, -NHCO-, -CONH-, -NHSO2-, -SON2NH-, -NHCONH-, -NHCSNH-, -CO-, -SO2-, -O-, -S-, pyrrolidine-2,5-dione, 1,2,3-triazole, -NH-, and -NMe-, wherein each p is independently 1 to 50.

[0346] In certain embodiments, the linking moiety of formula (LXVI) comprises a linear backbone of 6 to 40 consecutive atoms, for example, 10 to 40, 10 to 30, or 20 to 30 consecutive atoms.

[0347] In certain embodiments, the linking moiety of formula (LXVI) includes repeating ethylene glycol moieties (e.g., -CHCHO- or -OCHCH-). In certain cases, the linking moiety of formula (XVI) includes 2 to 20 ethylene glycol moieties, e.g., 2 to 15, 2 to 10, 3 to 20, 3 to 15, 3 to 10, 4 to 15, 5 to 15, or 5 to 10 ethylene glycol moieties. In some situations, the linking moiety of formula (XVI) includes two or more ethylene glycol moieties, e.g., 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, or even more ethylene glycol moieties.

[0348] In certain embodiments, the linking moiety of formula (LXVI) comprises one or more triazole linking moieties. In some cases, the linker comprises one or more 1,2,3-triazole linking moieties. In certain cases, the one or more 1,2,3-triazolyl moieties are selected from one of the following structures: [ka] In the formula, w1, u1, and q1 are independently 1 to 25 (e.g., 1 to 12, 1 to 6, etc.).

[0349] In certain embodiments, the linking moiety L 3 is (C 10 ~C 20 -Alkylene (e.g., C 12 -alkylene), or -(OCH2CH2) p -, where p is 1 to 25, for example, 3 to 25, 5 to 24, 7 to 25, 10 to 25, 15 to 25, or 20 to 24.

[0350] In some embodiments, the linker L has formula LXVII [ka] wherein: a is 0 to 12 (e.g., 2 to 6, or 2, or 3), b is 1 to 6 (e.g., 1, 2, or 3); c is 1 to 6 (e.g., 1, 2, or 3); r is 1 or 2, d is 1 to 6 (e.g., 1, 2, or 3); e is 1 to 6 (e.g., 1, 2, or 3); f is 1 to 6 (e.g., 1, 2, or 3); Z is the residue moiety resulting from the covalent bond between a chemoselective ligation group (eg, as described herein) of the linker precursor and a compatible group of Y.

[0351] In some embodiments of Formula LXVII, Z is a residue moiety resulting from the covalent bond (e.g., via a thioether bond) between a thiol-reactive chemoselective ligation group and one or more cysteine ​​residue(s) of Y. In some embodiments, the thiol-reactive chemoselective ligation group comprises a maleimide, a maleimide bromide, a haloacetamide, a vinyl sulfone, or a thiolactone. In some embodiments, the thiol-reactive group is selected from one of the following structures: [ka] During the ceremony, u is 1 to 11 (e.g., 1 to 5), v is 1 to 11 (e.g., 1 to 5), X is H or Br.

[0352] In some embodiments of Formula LXVII, Z is a residue moiety resulting from the covalent attachment (e.g., via an amide bond) of an amine-reactive chemoselective ligation group to one or more lysine residue(s) of Y. In some embodiments, the amine-reactive chemoselective ligation group comprises an active ester (e.g., N-hydroxysuccinimidyl (NHS) ester, sulfo-NHS ester, pentafluorophenyl (PFP) ester, tetrafluorophenyl (TFP) ester, etc.).

[0353] In some embodiments, the linker L is (LXVIIIa) to (LXVIIIc): [ka] wherein: a is 0 to 12 (e.g., 2 to 6, or 2, or 3), b is 1 to 6 (e.g., 1, 2, or 3); c is 1 to 6 (e.g., 1, 2, or 3); r is 1 or 2, d is 1 to 6 (e.g., 1, 2, or 3); e is 1 to 6 (e.g., 1, 2, or 3); f is 1 to 6 (for example, 1, 2, or 3).

[0354] In some embodiments of any one of Formulas (LXVII) or (LXVIIIa)-(LXVIIIc), a is 2 to 6, for example, 2 to 3. In some embodiments, a is 2. In some embodiments, a is 3. In some embodiments, a is 4. In some embodiments, a is 5. In some embodiments, a is 6.

[0355] In some embodiments of any one of Formulas (LXVII) or (LXVIIIa)-(LXVIIIc), b is 1 to 4, e.g., 1 to 3. In some embodiments, b is 1. In some embodiments, b is 2. In some embodiments, b is 3.

[0356] In some embodiments of any one of Formulas (LXVII) or (LXVIIIa) through (LXVIIIc), c is 1 to 4, e.g., 1 to 3. In some embodiments, c is 1. In some embodiments, c is 2. In some embodiments, c is 3.

[0357] In some embodiments of any one of Formulas (LXVII) or (LXVIIIa) through (LXVIIIc), r is 1. In some embodiments, r is 2.

[0358] In some embodiments of any one of Formulas (LXVII) or (LXVIIIa)-(LXVIIIc), d is 1 to 4, e.g., 1 to 3. In some embodiments, d is 1. In some embodiments, d is 2. In some embodiments, d is 3.

[0359] In some embodiments of any one of Formulas (LXVII) or (LXVIIIa) through (LXVIIIc), e is 1 to 5, e.g., 1 to 3. In some embodiments, e is 1. In some embodiments, e is 2. In some embodiments, e is 3. In some embodiments, e is 4. In some embodiments, e is 5.

[0360] In some embodiments of any one of Formulas (LXVII) or (LXVIIIa)-(LXVIIIc), f is 1 to 4, e.g., 1 to 3. In some embodiments, f is 1. In some embodiments, f is 2. In some embodiments, f is 3.

[0361] In some embodiments of any one of formulas (LXVII) or (LXVIIIa)-(LXVIIIc), a is 1-4, b is 1-4, c is 1-3, r is 1, d is 1-3, e is 1-6, and f is 1-3.

[0362] In some embodiments of any one of formulas (LXVII) or (LXVIIIa)-(LXVIIIc), a is 1-4, b is 1-4, c is 1-3, r is 2, d is 1-3, e is 1-6, and f is 1-3.

[0363] In some embodiments of any one of formulas (LXVII) or (LXVIIIa) through (LXVIIIc), a is 2, b is 1, c is 2, r is 1, d is 2, e is 3, and f is 2.

[0364] In some embodiments of any one of formulas (LXVII) or (LXVIIIa) through (LXVIIIc), a is 2, b is 1, c is 2, r is 2, d is 2, e is 3, and f is 2.

[0365] In some embodiments of any one of formulas (LXVII) or (LXVIIIa) through (LXVIIIc), a is 4, b is 1, c is 2, r is 1, d is 2, e is 3, and f is 2.

[0366] In some embodiments of any one of formulas (LXVII) or (LXVIIIa) through (LXVIIIc), a is 4, b is 1, c is 2, r is 2, d is 2, e is 3, and f is 2.

[0367] In some embodiments of any one of formulas (LXVII) or (LXVIIIa) through (LXVIIIc), a is 2, b is 2, c is 2, r is 1, d is 2, e is 3, and f is 2.

[0368] In some embodiments of any one of formulas (LXVII) or (LXVIIIa) through (LXVIIIc), a is 2, b is 2, c is 2, r is 2, d is 2, e is 3, and f is 2.

[0369] In some embodiments of any one of formulas (LXVII) or (LXVIIIa) through (LXVIIIc), a is 0, b is 3, c is 2, r is 2, d is 2, e is 3, and f is 2.

[0370] In some embodiments of any one of formulas (LXVII) or (LXVIIIa) through (LXVIIIc), a is 2, b is 4, c is 2, r is 2, d is 2, e is 3, and f is 2.

[0371] In some embodiments of any one of formulas (LXVII) or (LXVIIIa) through (LXVIIIc), a is 2, b is 4, c is 2, r is 1, d is 2, e is 3, and f is 2.

[0372] In some embodiments, the linker L is L A [ka] wherein Z 4 is selected from -NHC(O)NH-, -NHC(O)-, -C(O)NH-, -O-, -NH-; a is 0 to 12 (e.g., 2 to 6, or 2, or 3), b is 1 to 6 (e.g., 1, 2, or 3); c is 1 to 6 (e.g., 1, 2, or 3); d is 1 to 6 (e.g., 1, 2, or 3); e is 1 to 6 (e.g., 1, 2, or 3); f is 1 to 6 (for example, 1, 2, or 3).

[0373] L A In some embodiments, Z 4 is -NHC(O)NH-. In some cases, Z 4 is -NHC(O)-. In some cases, Z 4 is -C(O)NH-. In some cases, Z 4 is -O-. In some cases, Z 4 is -NH-.

[0374] L A In some embodiments, a is 1 to 4, b is 1 to 4, c is 1 to 3, d is 1 to 3, e is 1 to 6, and f is 1 to 3. In some embodiments, a is 4, b is 1, c is 2, d is 2, e is 5, and f is 2.

[0375] In some embodiments, Z 4 is -NHC(O)NH-, a is 1 to 4, b is 1 to 4, c is 1 to 3, r is 1, d is 1 to 3, e is 1 to 6, and f is 1 to 3. In some embodiments, Z 4 is -NHC(O)-, a is 1 to 4, b is 1 to 4, c is 1 to 3, r is 1, d is 1 to 3, e is 1 to 6, and f is 1 to 3.

[0376] In some embodiments, the linker L is L B [ka] wherein a is 0 to 12 (e.g., 2 to 6, or 2, or 3), b is 1 to 6 (e.g., 1, 2, or 3); c is 1 to 6 (e.g., 1, 2, or 3); r is 1 or 2, d is 1 to 6 (e.g., 1, 2, or 3); e is 1 to 6 (e.g., 1, 2, or 3); f is 1 to 6 (for example, 1, 2, or 3).

[0377] L B In some embodiments, a is 1 to 4, b is 1 to 4, c is 1 to 3, r is 1, d is 1 to 3, e is 1 to 6, and f is 1 to 3. In some embodiments, a is 4, b is 1, c is 2, r is 1, d is 2, e is 5, and f is 2. In some embodiments, a is 2, b is 1, c is 2, r is 1, d is 2, e is 3, and f is 2. In some embodiments, a is 4, b is 1, c is 2, r is 1, d is 2, e is 3, and f is 2. In some embodiments, a is 1, b is 2, c is 2, r is 1, d is 2, e is 3, and f is 2. In some embodiments, a is 0, b is 3, c is 2, r is 1, d is 2, e is 3, and f is 2.

[0378] L B In some embodiments, a is 1 to 4, b is 1 to 4, c is 1 to 3, r is 2, d is 1 to 3, e is 1 to 6, and f is 1 to 3. In some embodiments, a is 2, b is 1, c is 2, r is 2, d is 2, e is 3, and f is 2. In some embodiments, a is 4, b is 1, c is 2, r is 2, d is 2, e is 3, and f is 2. In some embodiments, a is 1, b is 2, c is 2, r is 2, d is 2, e is 3, and f is 2. In some embodiments, a is 0, b is 3, c is 2, r is 2, d is 2, e is 3, and f is 2.

[0379] In some embodiments, the linker L is L C [ka] wherein a is 0 to 12 (e.g., 1 to 6, 2 to 6, or 2, or 3); b is 1 to 6 (e.g., 1 to 4, 1, 2, or 3, etc.); c is 1 to 6 (e.g., 1 to 3, 1, 2, or 3, etc.); r is 1 or 2, d is 1 to 6 (e.g., 1 to 3, 1, 2, or 3, etc.); e is 1 to 6 (e.g., 1, 2, or 3); f is 1 to 6 (e.g., 1 to 3, 1, 2, or 3, etc.).

[0380] L C In some embodiments, a is 1 to 4, b is 1 to 4, c is 1 to 3, r is 1, d is 1 to 3, e is 1 to 6, and f is 1 to 3. In some embodiments, a is 2, b is 4, c is 2, r is 1, d is 2, e is 5, and f is 2.

[0381] L c In some embodiments, a is 1 to 4, b is 1 to 4, c is 1 to 3, r is 2, d is 1 to 3, e is 1 to 6, and f is 1 to 3. In some embodiments, a is 2, b is 4, c is 2, r is 2, d is 2, e is 5, and f is 2.

[0382] In certain embodiments of the ASGPR binding moiety (X) described herein, -Z 1 -L 1 - moiety (e.g., one of the linkers described herein). In some embodiments, the subject compounds are [ka] -Z comprising a linking moiety selected from 1 -L 1 - includes parts,

[0383] In the formula, each R 21 is independently selected from H, and optionally substituted (C-C) alkyl, and each R 22 are independently selected from H, halogen (e.g., F), and optionally substituted (C1-C6) alkyl, and o, p, q, r, s, t, u, v, w, x, y, z, and z1 are each independently 1 to 6.

[0384] In certain embodiments, Z 1 -L 1 -The group is [ka] and o is 1 or 2.

[0385] In certain embodiments, Z 1 -L 1 -The group is [ka] and each R 22 is H and p is 1 or 2.

[0386] In certain embodiments, Z 1 -L 1 -The group is [ka] wherein q is 1 to 3.

[0387] In certain embodiments, Z 1 -L 1 -The group is [ka] wherein r is 1 to 3.

[0388] In certain embodiments, Z 1 -L 1 -The group is [ka] wherein r is 1 to 3.

[0389] In certain embodiments, Z 1 -L 1 -The group is [ka] wherein s and t each independently represent an integer of 1 to 3.

[0390] In certain embodiments, Z 1 -L 1 -The group is [ka] wherein u is 1 to 3.

[0391] In certain embodiments, Z 1 -L 1 -The group is [ka] In the formula, v and w each independently represent an integer of 1 to 3.

[0392] In certain embodiments, Z 1 -L 1 -The group is [ka] In the formula, x is 0 to 3.

[0393] In certain embodiments, Z 1 -L 1 -The group is [ka] where y is 1 to 3.

[0394] In certain embodiments, Z 1 -L 1-The group is [ka] where R 21 is H and z is 1 to 4.

[0395] In certain embodiments, Z 1 -L 1 -The group is [ka] where R 21 is H, and z1 is 1 to 4.

[0396] In certain embodiments, Z 1 -L 1 -The group is [ka] and each R 22 is H and q is 1 to 3.

[0397] In certain embodiments, Z 1 -L 1 -The group is [ka] wherein q is 1 to 3.

[0398] In some embodiments, the subject compound is [ka] -Z comprising a linking moiety selected from 1 -L- group, where R 21 is independently selected from H, and optionally substituted (C-C) alkyl (e.g., methyl), and each R 22 is independently selected from H, halogen (e.g., F), and optionally substituted (C1-C6) alkyl (e.g., methyl). In certain embodiments, R 21is H. In certain embodiments, each R 22 is H.

[0399] In certain embodiments, -Z 1 -L 1 -The group is [ka] wherein q is 1 to 3. In certain cases, q is 1. In certain cases, q is 2. In certain cases, q is 3.

[0400] In certain embodiments, -Z 1 -L 1 -The group is [ka] is.

[0401] In certain embodiments, -Z 1 -L 1 - includes optionally substituted -NH-heteroarylene-. In certain embodiments, heteroarylene is triazole. In certain cases, heteroarylene is pyridine. In certain cases, heteroarylene is pyrimidine. In certain cases, heteroarylene is thiadiazole.

[0402] In certain embodiments, -Z 1 -L 1 -teeth, [ka] wherein R 24 and R 25 are H, optionally substituted C, (1~6) - independently selected from alkyl, optionally substituted fluoroalkyl, and halogen; 21 is independently selected from H, optionally substituted (C1-C6) alkyl, and optionally substituted alkanoyl. In certain cases, R21 is H. In certain cases, R 24 is C (1~3) -Alkyl, or C (1~3) -fluoroalkyl. In some cases, the fluoroalkyl is CF. In some cases, R 25 is C (1~3) -Alkyl, or C (1~3) -fluoroalkyl. Optionally, the fluoroalkyl is CF.

[0403] In some embodiments, the linker comprises a polypeptide scaffold, in which some or all of the side chain groups of the amino acid residues of such polypeptide scaffold have been modified to attach to X-linked moieties (e.g., as described herein). It is understood that the X-linked moieties (e.g., as described herein) can be conjugated to amino acid residues such as Asp, Lys, Orn, Glu, and Ser of the linker-containing polypeptide via any 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 backbone of such a linker can be a randomly polymerized polymer having an average length, or a polymer of a defined length, prepared, for example, in a controlled, stepwise manner. In some cases, the polypeptide linker has a length of 10 to 100 amino acid residues, e.g., 20 to 90, or 20 to 50 amino acid residues. In some embodiments, the N- or C-terminus of the polypeptide linker is modified to include a linking moiety to an additional X-linked 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 moieties (e.g., as described herein) suitable for attachment to a protein construct (Y) comprising a polypeptide that specifically binds to an autoantibody.

[0404] In some embodiments, the terms "linker," "linking moiety," and "linking group" are used interchangeably and refer to a linking moiety that covalently bonds 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 certain cases, the linker is a trivalent or higher polyvalent branched linking group. In some cases, the linker connecting two or more moieties has a linear or branched backbone with a length measured between the two or more moieties of 500 atoms or less (e.g., 400 atoms or less, 300 atoms or less, 200 atoms or less, 100 atoms or less, 80 atoms or less, 60 atoms or less, 50 atoms or less, 40 atoms or less, 30 atoms or less, or 20 atoms or less). The linking moiety can be a covalent bond connecting two groups, or a linear or branched chain 1 to 500 atoms in length, e.g., a chain of 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 linear, branched, cyclic, or single-atom. In certain cases, 1, 2, 3, 4, 5, or more, 10, or more, carbon atoms in the linker backbone can be optionally substituted with heteroatoms, such as sulfur, nitrogen, or oxygen heteroatoms. In certain situations, when the linker contains a PEG group, every third atom in that segment of the linker backbone is substituted with oxygen. The bonds between the backbone atoms can be saturated or unsaturated, and typically there are no more than one, two, or three unsaturated bonds in the linker backbone. The linker may include one or more substituents, such as, for example, an alkyl group, an aryl group, or an alkenyl group. Linkers include, but are not limited to, one or more of oligo(ethylene glycol), ether, thioether, disulfide, amide, carbonate, carbamate, tertiary amine, linear or branched alkyl (e.g., methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl), etc.). The linker backbone includes a cyclic group, such as, for example, an aryl, heterocycle, cycloalkyl group, or hetero group, wherein two or more atoms of the cyclic group, for example, 2, 3, or 4 atoms, are included in the backbone.

[0405] In some embodiments, a "linker" or linking moiety is derived from a molecule having two reactive ends, one for conjugation to a moiety of interest (Y), such as a biomolecule (e.g., an antibody), and one for conjugation to a moiety (designated X) that binds to a cell surface receptor (e.g., ASGPR). When Y is a polypeptide, the polypeptide-conjugation reactive end of the linker is a site that can be conjugated to a polypeptide via a cysteine ​​thiol or lysine amine group on the polypeptide, as the case may be, and thus may be a thiol-reactive group, such as 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.

[0406] 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 an amide bond, a urea or thiourea linkage, a carbamate linkage, an ester linkage, an amino linkage, an ether linkage, a thioether linkage, a sulfhydryl linkage, a heteroaryl linkage, or other heterofunctional linkage. 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. In certain cases, the linker includes one or more heteroaryl ring structures, for example, a triazole, such as a 1,2,3-triazole.

[0407] 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.

[0408] 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 and 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 and 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 400 Å and 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 200 Å and 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.

[0409] In certain embodiments, the linker L connects X and Y (or Z) by a chain of 4 to 500 contiguous atoms. 1In certain embodiments, the linker L separates X and Y (or Z) by a chain of 4 to 50 contiguous atoms. 1 In certain embodiments, the linker L separates X and Y (or Z) by a chain of 6 to 50 consecutive atoms, a chain of 11 to 50 consecutive atoms, a chain of 16 to 50 consecutive atoms, a chain of 21 to 50 consecutive atoms, a chain of 26 to 50 consecutive atoms, a chain of 31 to 50 consecutive atoms, a chain of 36 to 50 consecutive atoms, a chain of 41 to 50 consecutive atoms, or a chain of 46 to 50 consecutive 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 46-50 contiguous atoms. 1 ) and separate.

[0410] In certain embodiments, the linker L connects X and Y (or Z) by a chain of 4 or 5 consecutive atoms, a chain of 6-10 consecutive atoms, a chain of 11-15 consecutive atoms, a chain of 16-20 consecutive atoms, a chain of 21-25 consecutive atoms, a chain of 26-30 consecutive atoms, a chain of 31-35 consecutive atoms, a chain of 36-40 consecutive atoms, a chain of 41-45 consecutive atoms, or a chain of 46-50 consecutive atoms. 1 ) and separate.

[0411] 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.

[0412] 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 includes an alkylene, heteroatom, or optionally substituted heteroarylene linked to X.

[0413] 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 an optionally substituted triazole linked to X.

[0414] In some 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.

[0415] The linker is X to Z 1 It is understood that the linker may be considered to be directly connected to a group (e.g., as described herein). In some embodiments of any one of Formulas (Ia)-(Ip), the linker is Z1 Alternatively, -Z 1 -L 1 The - group (e.g., as described herein) can be considered part of the linking moiety connecting L to Y. The present disclosure is meant to include all such arrangements of X and linker (L).

[0416] In some embodiments of Formula (I), L is a group represented by Formula (II'''): [ka] wherein: L 1 and L 3 are independently linkers, and L 2 is a branched linking moiety, and in the formula, 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 Z 1 X's L via 1 represents the attachment point to *** represents the attachment point to Y, During the ceremony, If n is 1, a is 1 and b is 0, If n>1, a is 1 and b is 1.

[0417] In certain embodiments of the linker of formula (II), L 1 ~L 3 are -C respectively. 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 independently comprise 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 described herein), amino acid residue (natural or unnatural amino acid residue), -NH-, and -NMe-, wherein each p is independently 1 to 50.

[0418] 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) 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 situations, the linker of formula (II) 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.

[0419] In certain embodiments of the linker of formula (II), L 1 ~L 3 In some cases, the linker comprises one or more 1,2,3-triazole linking moieties. In certain cases, the one or more 1,2,3-triazolyl moieties are selected from one of the following structures: [ka] In the formula, w1, u1, and q1 are independently 1 to 25 (e.g., 1 to 12, 1 to 6, etc.).

[0420] In certain embodiments of a linker of formula (II), n is 1, such that b is 0, and the linker has formula (IIa'''): [ka] wherein: L 1 and L 3 are independently a linker (e.g., as described herein), and L 1 ~L 3 together provide a linear linker between X and Y, a is 1, c is 0 or 1, ** is Z 1 X's L via 1 represents the attachment point to *** represents the attachment point to Y.

[0421] In certain embodiments of the linker of Formula (IIa), the linear linker has a Z backbone, such as a backbone of 25 or more consecutive atoms, or 30 or more consecutive atoms, and in some cases up to 100 consecutive atoms. 1 In certain embodiments of Formula (IIa), the linear linker has a backbone of 20 or more consecutive atoms covalently linking X and Y (or Z) by a chain of 20 to 50 consecutive atoms. 1 In certain embodiments, the linear linker L separates X and Y (or Z) by a chain of 21 to 50 consecutive atoms, a chain of 22 to 50 consecutive atoms, a chain of 23 to 50 consecutive atoms, a chain of 24 to 50 consecutive atoms, a chain of 25 to 50 consecutive atoms, a chain of 26 to 50 consecutive atoms, a chain of 27 to 50 consecutive atoms, a chain of 28 to 50 consecutive atoms, or a chain of 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. 1In certain embodiments, a linear linker separates X and Y (or Z) by a chain of 31 to 60 contiguous atoms. 1 In certain embodiments, a linear linker separates X and Y (or Z) by a chain of 32 to 60 contiguous atoms. 1 In certain embodiments, a linear linker separates X and Y (or Z) by a chain of 33 to 60 contiguous atoms. 1 In certain embodiments, a 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 35-50 contiguous atoms. 1 In certain embodiments, the linear linker L separates X and Y (or Z) by a chain of 36 to 50 contiguous atoms. 1 In certain embodiments, the linear linker L separates X and Y (or Z) by a chain of 41 to 50 contiguous atoms. 1 In certain embodiments, the linear linker L separates X and Y (or Z) by a chain of 46-50 contiguous atoms. 1 ) and separate.

[0422] 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.

[0423] In some embodiments of the linker of formula (XI), n is 1, a is 1, b is 0, and c is 1, such that the linker L has the formula [ka] It is of the type.

[0424] In certain embodiments, the linear linker of formula (Xia) comprises a Z backbone, such as a backbone of 12 or more consecutive atoms, 14 or more consecutive atoms, or 16 or more consecutive atoms, and in certain embodiments, up to 100 consecutive atoms. 1 In certain embodiments of formula (Xia), the linear linker has a backbone of 10 or more consecutive atoms covalently linking X to Y via a chain of 20 to 50 consecutive atoms. 1 ) and Y. In certain embodiments of Formula (Xa), the linear linker L separates X (or Z) from Y by a chain of 30 to 60 contiguous atoms. 1 ) and Y.

[0425] In some embodiments of the linker of formula (XI), n is 2, a is 1, b is 1, and c is 1, such that the linker L has the formula (Xib): [ka] It is of the type.

[0426] In some embodiments of the linker of formula (XI), n is 3, a is 1, b is 1, and c is 1, such that the linker L has the formula (Xic): [ka] It is of the type.

[0427] In some embodiments of the linker of any one of formulas (XI) or (Xia) through (Xic), each L 1 is represented by formula (XII) [ka] wherein: L 10 is the linking part, * is Z 1 X's L via 1 represents the attachment point to L 11 ~L 19 is independently absent or a linking moiety, In the formula, each L 1 L 10 ~L 19 -C 1~6 -Alkylene-, -C 1~12 -Alkylene-, -C 1~20 -Alkylene-, -NHCO-C 1~6 -Alkylene-, -CONH-C 1~6 -Alkylene-, -NH-C 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-, -CC 1~6 -Alkylene-NHCSNH-, -O(CH2) p -, -(OCH2CH2) p -, -NHCO-, -CONH-, -NHSO2-, -SO2NH-, -NHCONH-, -NHCSNH-, -CO-, -SO2-, -O-, -S-, arylene, heteroarylene, heteroalkylene, cycloalkylene, -NH-, -NC 1~6 -N(CH3)-, and -N(CH3)-, wherein each L 1 L 10 ~L 19 are each independently substituted with one or more halo (e.g., 1 to 3, or 1 to 5), and p is independently 1 to 50, e.g., 1 to 20, 1 to 12, 1 to 10, 1 to 8, or 1 to 6, e.g., 1, 2, 3, 4, 5, or 6.

[0428] In some embodiments of the linker of any one of formulas (XI) or (Xia) through (Xic), each L 1 is represented by formula (XII) [ka] wherein: L 10 is the linking part, * is Z 1 X's L via1 represents the attachment point to L 11 ~L 19 is independently absent or a linking moiety, In the formula, each L 1 L 10 ~L 19 -C 1~6 -Alkylene-, -CF2-, -C 1~12 -Alkylene-, -C 1~20 -Alkylene-, -NHCO-C 1~6 -Alkylene-, -CONH-C 1~6 -Alkylene-, -NH-C 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 -, -NHCO-, -CONH-, -NHSO2-, -SO2NH-, -NHCONH-, -NHCSNH-, -CO-, -SO2-, -O-, -S-, pyrrolidine-2,5-dione, 1,2,3-triazole, -NH-, -N(C 1~6 -alkyl)-, and -N(CH3)-, where each p is independently 1 to 50, e.g., 1 to 20, 1 to 12, 1 to 10, 1 to 8, or 1 to 6, e.g., 1, 2, 3, 4, 5, or 6.

[0429] In certain embodiments of Formula (XII), the linking moiety L 1 comprises a linear backbone of 6 to 40 consecutive atoms, e.g., 10 to 40, 10 to 30, 16 to 30, or 20 to 30 consecutive atoms. In certain embodiments of Formula (XII), the linking moiety L 1comprises a linear backbone of 6 to 20 consecutive atoms, for example 6 to 16 consecutive atoms, for example 8, 9, 10, 11, 12, 13, 14, 15, or 16 consecutive atoms.

[0430] In certain embodiments, the linking moiety of formula (XII) comprises one or more repeating ethylene glycol moieties (e.g., -CH2CHO- or -OCH2CH2-). In certain embodiments, the linking moiety of formula (XII) comprises 1 to 10 ethylene glycol moieties, e.g., 1, 2, 3, 4, 5, or 6 ethylene glycol moieties.

[0431] In certain embodiments, the linking moiety of formula (XII) comprises one or more triazole (e.g., 1,2,3-triazole) containing linking moieties, which can be derived from azide-alkyne click chemistry and thus have two possible directions depending on the synthetic method.

[0432] In certain embodiments of Formula (II'''), n is 2 or greater and L 2 teeth, [ka] wherein x and y are each independently 1 to 10.

[0433] In certain embodiments of Formula (II'''), L 1 ~L 2 comprises a backbone 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.

[0434] In certain embodiments of formula (II''') or (IIa), L 3 comprises a backbone of 10 to 80 consecutive atoms, such as 12 to 70, 12 to 60, or 12 to 50 consecutive atoms. In some embodiments, L comprises 12 to 70, 12 to 60, 12 to 50, or 10 to 60 consecutive straight or branched chain atoms.

[0435] 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, for example, 3 to 25, 5 to 24, 7 to 25, 10 to 25, 15 to 25, or 20 to 24.

[0436] In certain embodiments, L is a group of formula (IIb): [ka] wherein: L 1 ~L 5 are each independently a linking moiety, and together they form Z 1 and Y, forming a linear or branched linker therebetween; a, b, c, d, and e are each independently 0, 1, or 2; ** is Z 1 X's L via 1 represents the attachment point to *** represents the attachment point to Y, During the ceremony, If n is 1, a is 1 and c is 0, If n>1, a is 1 and c is 1.

[0437] In some embodiments, L is a group of formula (IIb'): [ka] wherein: L 1 ~L 6 are each independently a linking moiety, and together they form Z 1 and Y, forming a linear or branched linker therebetween; a, b, c, d, and e are each independently 1, 2, 3, 4, or 5; ** is Z1 X's L via 1 represents the attachment point to *** represents the attachment point to Y.

[0438] In certain embodiments of the linker of formula (IIb), L 1 ~L 5 are -C respectively. 1~20 -Alkylene-, -NHCO-C 1~6 -Alkylene-, -CONH-C 1~6 -Alkylene-, -NH C 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 independently comprise 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 described herein), amino acid residue (natural or unnatural amino acid residue), -NH-, and -NMe-, wherein each p is independently 1 to 50.

[0439] In certain embodiments, L is a group of formula (IIb'): [ka] wherein: L 1 ~L 6 are each independently a linking moiety, and together they form Z 1and Y, forming a linear or branched linker therebetween; a, b, c, d, and e are each independently 1, 2, 3, 4, or 5; ** is Z 1 X's L via 1 represents the attachment point to *** represents the attachment point to Y.

[0440] In certain embodiments, each L 1 ~L 5 -C 1~20 -Alkylene-, -NHC(O)-C 1~6 -Alkylene-, -C(O)NH-C 1~6 -Alkylene-, -NH-C 1~6 -Alkylene-, -NHC(O)NH-C 1~6 -Alkylene-, -NHC(S)NH-C 1~6 -Alkylene-, -C 1~6 -Alkylene-NHC(O)-, -C 1~6 -Alkylene-C(O)NH-, -C 1~6 -Alkylene-NH-, -C 1~6 -Alkylene-NHC(O)NH-, -C 1~6 -Alkylene-NHC(S)NH-, -O(CH2) p -, -(OCH2CH2) p and independently comprise one or more linking moieties independently selected from -, -NHC(O)-, -C(O)NH-, -NHS(O)2-, -S(O)2NH-, -C(O)-, -S(O)2-, -O-, -S-, monocyclic heteroaryl, monocyclic aryl, monocyclic heterocycle, monocyclic carbocycle, an amino acid residue, -NH-, and -NMe-, wherein L 1 ~L 5 are each independently optionally substituted with 1 to 5 halo; each p is independently 1 to 50; L 6 -C 1~20 -Alkylene-, -NR 16 C(O)-C 1~6 -Alkylene-, -C(O)NR 16 -C 1~6 -Alkylene-, -NR 16 -C1~6 -Alkylene-, -NR 16 C(O)NR 16 -C 1~6 -Alkylene-, -NR 16 C(S)NR 16 -C 1~6 -Alkylene-, -C 1~6 -Alkylene-NR 16 C(O)-, -C 1~6 -Alkylene-C(O)NR 16 -, -C 1~6 -Alkylene-NR 16 -, -C 1~6 -Alkylene-NR 16 C(O)NR 16 -, -C 1~6 -Alkylene-NR 16 C(S)NR 16 -, -O(CH2) p -, -(OCH2CH2) p -, -NR 16 C(O)-, -C(O)NR 16 a linking group comprising one or more linking moieties independently selected from -, -NHS(O)2-, -S(O)2NH-, -C(O)-, -S(O)2-, -O-, -S-, monocyclic heteroaryl, monocyclic aryl, monocyclic heterocycle, an amino acid residue, or -NR16-; Each R 16 is independently —H, (C1-C6) alkyl, or monocyclic heteroaryl.

[0441] In certain embodiments, each L 1 ~L 5 -C 1~20 -Alkylene-, -NHC(O)-C 1~6 -Alkylene-, -C(O)NH-C 1~6 -Alkylene-, -NH-C 1~6 -Alkylene-, -NHC(O)NH-C 1~6 -Alkylene-, -NHC(S)NH-C 1~6 -Alkylene-, -C 1~6 -Alkylene-NHC(O)-, -C 1~6 -Alkylene-C(O)NH-, -C 1~6 -Alkylene-NH-, -C1~6 -Alkylene-NHC(O)NH-, -C 1~6 -Alkylene-NHC(S)NH-, -O(CH2) p -, -(OCH2CH2) p -, -NHC(O)-, -C(O)NH-, -NHS(O)2-, -S(O)2NH-, -C(O)-, -S(O)2-, -O-, -S-, monocyclic heteroaryl, monocyclic aryl, monocyclic heterocycle, monocyclic carbocycle, an amino acid residue, -NH-, and -NMe-; 1 ~L 5 are each independently optionally substituted with 1 to 5 halo; each p is independently 1 to 50; L 6 teeth, [ka] where R z teeth, [ka] is.

[0442] In certain embodiments of Formula (IIb), -(L 1 ) a - includes an optionally substituted alkyl or ethylene glycol linking moiety. In certain cases, L 1 is an optionally substituted -C 1~6 -alkylene. In certain cases, L 1 contains an ethylene glycol linking moiety.

[0443] In certain embodiments of Formula (IIb), L 1 teeth, -C 1~6 -Alkylene-, -(CH2CH2O) t -, --C 1~6 -Alkylene-NR 4 CO-, -C 1~6 -alkyleneCONH-, or OCH2, where t is 1 to 20; 4is independently selected from H and optionally substituted (C1-C6) alkyl. In certain cases, L 1 -C 1~6 -Alkylene-, e.g., --C 1~3 -alkylene-. In certain cases, L 1 is -(CH2CH2O) t -, where t is 1 to 20, for example, 1 to 15, 1 to 10, 1 to 8, 1 to 6, or 1 to 4. In certain cases, L 1 is --C 1~6 -Alkylene-NR 4 CO-. In certain cases, L 1 -C 1~6 -alkyleneCONH-. In certain cases, L 1 is OCH2.

[0444] In some embodiments of Formula (IIb), one or more L 1 are independently -CH2O-, -(CH2CH2O) t -, -NR 4 CO-, -C 1~6 -alkylene-, [ka] where 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 1 Any of the moieties are optionally further substituted.

[0445] In certain embodiments of Formula (IIb), L 2 teeth, -NR 4’ CO-C 1~6 -Alkylene-, -CONR4’ -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. 2 is -NR 4’ CO-C 1~6 -alkylene-. In certain cases, L 2 -CONR 4’ -C 1~6 - alkylene.

[0446] In certain cases, L 2 teeth, [ka] wherein w is 1 and u is 0 or 1.

[0447] In certain cases, L 2 teeth, [ka] wherein w is 1 and u is 0 or 1.

[0448] In certain cases, L 2 teeth, [ka] wherein w is 1, u is 0 or 1, and q is 1.

[0449] In certain cases, L 2 teeth, [ka] where u is 0 or 1.

[0450] In certain cases, L 2 teeth, [ka] is.

[0451] 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, 2 to 4.

[0452] In certain embodiments of Formula (IIb), L 4 does not exist or -C 1~6 -Alkylene-, -(CH2CH2O) t -, -C 1~6 -Alkylene-NHCO-, -C 1~6 -alkyleneCONH-, or OCH2. In certain cases, L 4 does not exist. In certain cases, L 4 -C 1~6 -alkylene-. In certain cases, L 4 is -(CH2CH2O) t -, where 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 is --C 1~6 -alkylene-NHCO-. In certain cases, L 4 -C 1~6 -alkyleneCONH-. In certain cases, L 4 is OCH2.

[0453] In some embodiments of the subject compounds, n is 1 and L in formula (IIb) 3does not exist.

[0454] In certain embodiments of the subject compounds, n is 2 or greater, and L in formula (IIb) 3 is a branched linking moiety.

[0455] Thus, in some embodiments of formula (IIb), L 3 is a branched linking moiety, e.g., a bivalent or trivalent linking moiety. For example, L 3 The linking moiety may be one of the following general formulas: [ka]

[0456] In some embodiments of Formula (IIb), the branched linking moiety has a higher valence and can be described by one of the following general formulas: [ka] etc. In the formula, any two L 3 The groups can be directly linked or connected via any linear linking moiety (eg, as described herein).

[0457] In some embodiments of Formula (IIb), the branched linking moiety comprises one, two, or more L, each of which is a trivalent moiety. 3 Linking moieties, which when linked together provide multiple branching points for covalent attachment of ligands, can be represented by the general formula: [ka] In the formula, t is 0 to 500, for example, 0 to 100, 0 to 20, or 0 to 10.

[0458] In some embodiments, a branched linking moiety (e.g., L 3) includes one or more of 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.

[0459] In some embodiments of Formula (IIb), one or more L 3 teeth, [ka] a branched moiety selected from wherein each x and y is independently 1 to 10, 1 to 6, 1 to 3, etc., for example, 1 or 2. In some cases, each x is 1, 2, or 3, for example, 2.

[0460] In some embodiments of Formula (IIb), L 5 are independently -CH2O-, -(CH2CH2O) t -, -NR 4 CO-, -C 1~6 -alkylene-, [ka] is selected from: 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 are optionally further substituted.

[0461] In certain cases, L 5 is -CH2O-. In certain cases, L 5 is -(CH2CH2O) t-, where t is 1 to 20, for example, 1 to 15, 1 to 12, 1 to 10, 1 to 8, 1 to 6, or 1 to 4. In certain cases, L 5 is -NR 4 CO—, where R 4 is H or optionally substituted (C1-C6) alkyl. In certain cases, L 5 -C 1~6 -alkylene-.

[0462] In certain cases, L 5 teeth, [ka] In the formula, r is 0 to 20, for example, 0 to 15, 0 to 10, 0 to 8, or 0 to 5.

[0463] 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.

[0464] 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; 13 is H or optionally substituted (C1-C6) alkyl.

[0465] 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; 13is H or optionally substituted (C1-C6) alkyl.

[0466] 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; 13 is H or optionally substituted (C1-C6) alkyl.

[0467] 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.

[0468] 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.

[0469] 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.

[0470] 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.

[0471] In certain cases, L 5 teeth, [ka] In the formula, r is 0 to 20, for example, 0 to 15, 0 to 10, 0 to 8, or 0 to 5.

[0472] In some embodiments of Formula (IIb), L 5 comprises one or more of 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), and the like. Amino acid analogs include, but are not limited to, unnatural amino acids, as well as other modifications known in the art. Amino acids include 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.

[0473] In some embodiments of Formula (IIb), L 1 ~L 5 is the following unit

[0474] [ka] wherein R a is (C1-C6) alkyl or substituted (C1-C6) alkyl, for example, (C1-C6) alkyl optionally substituted with an amine, a tertiary amine, an optionally substituted alkoxy, an optionally substituted carboxyl, an optionally substituted aryl, or an optionally substituted heteroaryl. R a It is understood that can be linked to the M6PR binding moiety.

[0475] In some embodiments, the linker comprises a polypeptide scaffold in which some or all of the side groups of the amino acid residues have been modified to attach X-linking moieties (e.g., as described herein). It is understood that the X-linking moieties (e.g., as described herein) can be conjugated to amino acid residues such as Asp, Lys, Orn, Glu, and Ser of the linker-containing polypeptide via convenient conjugation chemistry. In some embodiments, the linker contains a polylysine polypeptide. In some embodiments, the linker contains a polyornithine polypeptide. In some embodiments, the linker contains a polyserine polypeptide. In some embodiments, the linker contains a polyaspartic acid polypeptide. The polypeptide may be a randomly polymerized polymer having an average length, or a polymer of a defined length, prepared, for example, in a controlled, stepwise manner. In some cases, the polypeptide linker segment has a length of 10 to 100 amino acid residues, e.g., 20 to 90, or 20 to 50 amino acid residues. In some embodiments, the N-terminus 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 suitable for attachment to a Yi moiety of interest (e.g., as described herein).

[0476] 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.

[0477] In certain embodiments of Formula (II), (IIa), or (IIb), the linker comprises 20 to 100 consecutive atoms, such as 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, such as 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.

[0478] In certain embodiments of Formula (II), (IIa), or (IIb), the linker comprises 25 or more consecutive atoms, such as 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, such as 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 more consecutive atoms.

[0479] It is believed that the compound of the present disclosure, which has a specific configuration with a linker of desired valency and length, can simultaneously specifically bind to both receptor and target with high affinity, and show high target uptake activity.Therefore, the conjugate of the present disclosure can sequester target protein in cell lysosome and degrade target protein.

[0480] Exemplary Linkers and Linking Moieties Exemplary linkers and linking moieties that can be utilized in preparing compounds of the present disclosure are shown in Tables 6-8.

[0481] In certain embodiments, the linker is a linear linker or linking moiety as shown in Table 6. [Table 6-1] [Table 6-2] [Table 6-3]

[0482] Table 7 includes various linker component synthetic precursors (eg, linear and branched linker precursors) that can be utilized to prepare the subject compounds. [Table 7-1] [Table 7-2] [Table 7-3] [Table 7-4] [Table 7-5] [Table 7-6] [Table 7-7] [Table 7-8] [Table 7-9] [Table 7-10]

[0483] In certain embodiments, the linker is a branched linker or linking moiety as shown in Table 8. [Table 8-1] [Table 8-2] [Table 8-3] [Table 8-4] [Table 8-5] [Table 8-6]

[0484] Table 9 provides exemplary synthetic precursors of linker components used to prepare compounds of the present disclosure, for example, via conjugation chemistry. It is understood that various homologs of the structures shown in Table 9 are also included in the present disclosure, providing linkers of various lengths. It is understood that alternative chemoselective ligation groups and other chemical functionalities can also be incorporated as needed to prepare desired linkers. [Table 9-1] [Table 9-2] [Table 9-3] [Table 9-4] [Table 9-5] [Table 9-6] [Table 9-7] [Table 9-8]

[0485] Chemoselective Ligation Groups A chemoselective ligation group is a group having a reactive functional group or group that can be conjugated with a compatible group on a second moiety. For example, the chemoselective ligation group (or its precursor) 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-Picteth-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, dialkylsquarate chemistry, etc.

[0486] Chemoselective ligation groups available for joining two moieties include, but are not limited to, amino (e.g., the N-terminal amino group or lysine side chain group of a polypeptide), azide, aryl azide, alkynyl (e.g., ethynyl or cyclooctyne or derivative), 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, cyanoalkyne, thiol (e.g., cysteine ​​residue), disulfide or protected thiol, isocyanate, isothiocyanate, aldehyde, ketone, alkoxyamine, hydrazide, aminooxy, phosphine, HPS hydrazinyl-indolyl group, or aza-HPS hydrazinyl-pyrrolo-pyridinyl group, tetrazine, cyclooctene, squarate, and the like.

[0487] In some situations, a chemoselective ligation group can spontaneously conjugate to a compatible chemical group when the two groups are contacted under appropriate conditions (e.g., copper-free click chemistry conditions). In some situations, a chemoselective ligation group can conjugate to a compatible chemical group when the two groups are contacted in the presence of a catalyst or other reagent (e.g., copper-catalyzed click chemistry conditions).

[0488] In some embodiments, the chemoselective ligation group is a photoactive ligation 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.

[0489] In some situations, Y is a reactive functional group or a precursor of a functional group capable of conjugating with a compatible group on a second moiety, for example, a carboxylic acid is a precursor of an active ester chemoselective ligation group.

[0490] In certain embodiments of Formula (I), Y is a reactive moiety that can form a covalent bond with a polypeptide (e.g., with an amino acid side chain of the polypeptide that has a compatible reactive group). The reactive moiety can be referred to as a chemoselective ligation group.

[0491] In certain embodiments of Formula (I), Y is a thio-reactive chemoselective ligation group (e.g., as described in Table 10). In some cases, Y can generate a residue moiety Z resulting from the covalent attachment of a thiol-reactive chemoselective ligation group to one or more cysteine ​​residue(s) of a protein, e.g., an Ab.

[0492] In certain embodiments of Formula (I), Y is a Cys-reactive chemoselective ligation group (e.g., a maleimide derivative described in Table 10). In some cases, the Cys-reactive chemoselective ligation group comprises a maleimide group. In some embodiments, the chemoselective ligation group comprises a maleimide group from Table 22, e.g., mal-1 through mal-7.

[0493] In certain embodiments of Formula (I), Y is an amino-reactive chemoselective ligation group (e.g., as described in Table 10). In some cases, Y can generate a residue moiety Z obtained by covalently attaching the amine-reactive chemoselective ligation group to one or more lysine residue(s) of a protein, e.g., an Ab.

[0494] In certain embodiments of Formula (I), Y is a Lys-reactive chemoselective ligation group (e.g., an active ester described in Table 10). In some embodiments, the Lys-reactive chemoselective ligation group is a PFP ester.

[0495] Exemplary chemoselective ligation groups, and their synthetic precursors, that can be adapted for use in the compounds of the present disclosure are shown in Table 10. [Table 10-1] [Table 10-2] [Table 10-3] [Table 10-4] [Table 10-5]

[0496] In Table 10, [ka] can represent the linking moiety of Y or the point of attachment to the linked X moiety.

[0497] Table 11 shows exemplary residue moieties, where "***" indicates the attachment point of Y. [Table 11-1] [Table 11-2]

[0498] Exemplary ASGPR-binding compounds with chemoselective ligation groups for preparing conjugates The present disclosure provides: (1) one or more specific ASGPR ligands (X) (e.g., as described herein, e.g., ligands X1-X20 in Tables 1-5) 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 described herein, e.g., in Tables 6-8), and (3) Compounds of formula (I) include those that can be prepared from precursor ligand linker compounds that include a chemoselective ligation group (Y), such as those described herein, e.g., any one of the groups in Table 10.

[0499] Table 12 shows various monovalent ASGPR ligand linker compounds for use in preparing the conjugates of the present disclosure. [Table 12-1] [Table 12-2]

[0500] Table 13 shows various multivalent ASGPR ligand linker compounds for use in the conjugates of the present disclosure. [Table 13-1] [Table 13-2]

[0501] Tables 14-17 show several exemplary ASGPR-binding compounds of the present disclosure that include a chemoselective ligation group or precursor thereof. It is understood that the present disclosure includes Y (e.g., as described herein) conjugates of each exemplary compound in Tables 14-17. For example, conjugates are included in which the chemoselective ligation group is conjugated to a different Y, such as an antibody or antibody fragment against a target protein.

[0502] The chemoselective ligation group of such compounds can be utilized to connect to another Y moiety of interest (e.g., as described below). It will be appreciated that any of these compounds can also be prepared de novo to include an alternative Y moiety of interest (e.g., as described below) rather than the chemoselective ligation group. In some embodiments, such compounds are referred to as conjugates, such as biomolecular conjugates that specifically bind to target proteins. [Table 14-1] [Table 14-2] [Table 14-3] [Table 14-4] [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 15-18 Table 15-19 Table 15-20 [Table 15-21] [Table 15-22] [Table 15-23] [Table 15-24] [Table 15-25] [Table 15-26] [Table 16-1] [Table 16-2] [Table 17-1] [Table 17-2]

[0503] The present disclosure is intended to encompass any one stereoisomer of the compounds described herein. In some circumstances, the compounds include enantiomers of DN-acetylgalactosamine (GalNAc), or analogs or derivatives of GalNAc.

[0504] 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]

[0505] 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] [Table 19-10] [Table 19-11] [Table 19-12]

[0506] Table 20 shows exemplary monovalent ASGPR-binding intermediate compounds of the present disclosure that include a promoiety and an X group of formula (Ib). [Table 20]

[0507] Table 21 shows exemplary ASGPR-binding intermediate compounds of the present disclosure that include an X group of formula (In). [Table 21]

[0508] Table 22 shows exemplary ASGPR binding intermediate compounds. [Table 22-1] [Table 22-2] [Table 22-3] [Table 22-4] [Table 22-5] [Table 22-6] [Table 22-7] [Table 22-8] [Table 22-9] [Table 22-10] [Table 22-11] [Table 22-12] [Table 22-13] [Table 22-14] Table 22-15 Table 22-16 Table 22-17 Table 22-18 Table 22-19 Table 22-20 Table 22-21 Table 22-22 Table 22-23 Table 22-24 Table 22-25 Table 22-26 Table 22-27 Table 22-28 Table 22-29 Table 22-30

[0509] The present disclosure is intended to encompass any one stereoisomer of the compounds described herein. In some circumstances, the compounds include enantiomers of DN-acetylgalactosamine (GalNAc), or analogs or derivatives of GalNAc.

[0510] Exemplary CI-M6PR Binding Compounds with Chemoselective Ligation Groups for Preparing Conjugates Exemplary M6PR binding moieties X of Formulas (I)-(XIII) that can be used to prepare compounds and conjugates of the present disclosure are shown in Table 23. [Table 23-1] [Table 23-2] [Table 23-3] [Table 23-4] [Table 23-5] [Table 23-6] [Table 23-7] [Table 23-8]

[0511] Exemplary synthons or synthetic precursors that can be utilized in the preparation of compounds of the present disclosure to incorporate a desired M6PR binding moiety are shown in Table 24. It is understood that depending on the M6PR binding moiety and linker selected, alternative synthons are possible, including homologs and analogs of those shown in Table 24. The synthons in Table 24 are linked to a linking moiety Z 3and structural elements that become part of the linker (L) in the compounds and conjugates of the present disclosure. Based on the exemplary synthetic precursors in Table 24, it is understood that synthons corresponding to any of the M6PR binding moieties in Table 23 can be utilized to prepare compounds of the present disclosure. [Table 24-1] [Table 24-2] [Table 24-3] [Table 24-4] [Table 24-5] [Table 24-6] [Table 24-7] [Table 24-8] [Table 24-9] [Table 24-10]

[0512] Other M6PR binding moieties of interest and their synthons or synthetic precursors are shown in Table 25. X101-X103 represent compounds with a phosphate or thiophosphate head group. X109-X110 represent exemplary compounds of formula (V). In some embodiments, such M6PR binding moieties are used in reference compounds for evaluation of compounds of formula (XII). [Table 25-1] [Table 25-2]

[0513] Conjugates A compound of the present disclosure can be referred to as a conjugate, for example, when the moiety of interest (Y) is an antibody or antibody fragment (e.g., as described herein). In one embodiment, the conjugate comprises a ligand moiety conjugated to a target binding site via a linker, wherein the ligand moiety binds to a lysosomal targeting molecule extracellularly, the target binding moiety binds to the target molecule extracellularly, the target binding moiety dissociates from the target molecule in an endosome, and the conjugate is externalized from the cell. Such a conjugate can be prepared by conjugating a chemoselective ligation group of any one of the compounds described herein with a compatible reactive group on molecule Y. The compatible group on molecule Y can be introduced by modification prior to conjugation or can be a group present within the molecule. Alternatively, such a conjugate can be prepared de novo, for example, by modifying the molecule of interest Y as a starting material and introducing a linker to which, for example, a ligand or lysosomal targeting molecule binding moiety (X) can be attached.

[0514] In some embodiments, the moiety of interest to which the lysosomal targeting molecule binding moiety is attached is a biomolecule. In some embodiments, the moiety of interest is a biomolecule. In some embodiments, Y is a biomolecule that specifically binds to a target molecule, such as a target protein. In some embodiments, the biomolecule is an antibody or an antibody fragment.

[0515] In some embodiments, the desired portion is a molecule that specifically binds to a desired target, i.e., a target binding portion.In this case, the conjugate of the present disclosure provides for the target to be taken up into cells and / or degraded after the target is non-covalently bound to the conjugate.The present inventors have demonstrated that the conjugate of the present disclosure has a specific arrangement of a lysosome targeting molecule binding portion with desired affinity and a linker with desired valency and length, which can specifically bind to both the lysosome targeting molecule and the target at the same time with high affinity.Therefore, the conjugate of the present disclosure can sequester target protein in the lysosome of cells and degrade the target protein.

[0516] The compounds of the present disclosure may also be referred to as conjugates in some cases, 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 ligation 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 that is 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 ligation chemistry.

[0517] Aspects of the present disclosure include compounds of formula (I) in which 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. In some embodiments, the conjugate comprises a region of interest Y that specifically binds to the target protein and can be used in a method of cellular uptake or internalization of the target protein via binding to a cell surface receptor and eventual degradation of the target protein.

[0518] In one embodiment, the conjugate promotes degradation of the target and can repeatedly "cycle" in and out of the cell. It is hypothesized that one of two cycling mechanisms occurs, although in some cases, both mechanisms may occur for a given conjugate. Referring to FIG. 1A, conjugate 10 includes a target-binding moiety 11, a linker 12, and a ligand moiety 13. In step A, conjugate 10 binds to a lysosomal targeting molecule 20 extracellularly and also to a target 30 extracellularly. Such binding can occur in any temporal order. While target 30 is depicted as a soluble target, it may also be a transmembrane target or other target, as described herein. The resulting ternary conjugate is internalized into intracellular vesicles (step B), forming endosomes 40 in step C. Within endosomes 40, a first cycling mechanism occurs, in which events (i) the conjugate dissociates from the lysosomal targeting molecule, (ii) the conjugate dissociates from the target, and (iii) the conjugate binds to FcRn50. Events (i), (ii), and (iii) can occur in any temporal order. As shown in steps D1, E1, and F, the conjugate and target have distinct fates: in step D1, the conjugate remains in the endosome; in step E1, it is externalized from the cell; and in step F, the target is degraded in lysosomes 60.

[0519] Figure 1B shows some of the same components, functions, and steps as shown in Figure 1A. However, in this second cycling mechanism, the conjugate remains bound to the lysosomal targeting molecule 20 in the endosome 40 and is externalized from the cell in steps D2 and E2. The target is degraded in the lysosome 60 in step F, as in the previous mechanism.

[0520] Using these mechanisms, the conjugates of the present disclosure "cycle" (internalize and externalize) inside and outside the cell to promote lysosomal degradation of targets. Such cycling allows for sustained activity on the order of hours to days, allowing a single conjugate to promote lysosomal degradation of multiple targets. In other words, the conjugates described herein can be used to repeatedly bind to targets, internalize with them to promote lysosomal degradation, and return to the cell membrane to bind additional targets, thereby degrading superstoichiometric ratios of targets.

[0521] In some embodiments, for example, after the conjugate binds to a lysosomal transport receptor, the conjugate is taken up into the cell via a lysosomal targeting molecule. Receptor-mediated internalization is described, for example, in G.Ahn et al., Nat.Chem.Biol.2021,17(9)937-46 and the references cited therein. As described elsewhere herein, in various embodiments, the lysosomal targeting molecule is ASGPR. In other embodiments, the lysosomal targeting molecule is M6PR. In other embodiments, the lysosomal targeting molecule is LDLR or CD63.

[0522] In some embodiments, the ligand moiety X remains bound to the lysosomal targeting molecule within the endosome. In some embodiments, the conjugate is externalized from the cell via the lysosomal targeting molecule. In some embodiments, the ligand moiety has equal binding affinity for the lysosomal targeting molecule extracellularly and within the endosome. In some embodiments, the ligand moiety X has equal binding affinity for the lysosomal targeting molecule at extracellular pH and intraendosomal pH. In some embodiments, the ligand moiety X binds to extracellular Ca 2+ Concentration and intraendosomal Ca 2+ have equal binding affinity for the lysosomal targeting molecule at different concentrations.

[0523] Externalization of the conjugate can also be mediated by FcRn. In some embodiments, for example, after target binding moiety Y binds to FcRn, the conjugate is externalized from the cell via FcRn. In some embodiments, ligand moiety X binds to FcRn within an endosome. In some embodiments, the conjugate dissociates from the lysosomal targeting molecule within an endosome. In some embodiments, ligand moiety X has a higher binding affinity for the lysosomal targeting molecule outside the cell than within the endosome. In some embodiments, ligand moiety X has a higher binding affinity for the lysosomal targeting molecule at extracellular pH than at intraendosomal pH. In some embodiments, ligand moiety X binds to intraendosomal Ca. 2+ Extracellular Ca concentration 2+ The ligand moiety X has high binding affinity to the lysosome-targeting molecule at high concentrations. In some embodiments, the ratio of the binding affinity between the ligand moiety X and the lysosome-targeting molecule within the endosome to the extracellular region is 1:2 to 2:1. In some embodiments, the ratio of the binding affinity between the ligand moiety X and the lysosome-targeting molecule within the endosome to the extracellular region is 1:5 to 5:1. In some embodiments, the ratio of the binding affinity between the ligand moiety X and the lysosome-targeting molecule within the endosome to the extracellular region is 1:10 to 10:1.

[0524] In some embodiments, the target-binding moiety Y has a higher binding affinity for FcRn in endosomes than extracellularly. In some embodiments, the ratio of the binding affinity of the target-binding moiety Y to FcRn in endosomes to extracellularly is 1:100 to 1:10. In some embodiments, the target-binding moiety Y has a higher binding affinity for FcRn at endosomal pH than at extracellular pH. In some embodiments, Y has enhanced binding to FcRn compared to wild-type at endosomal pH. In some embodiments, the target-binding moiety Y has approximately the same binding affinity for FcRn extracellularly as the binding affinity that wild-type IgG has for FcRn extracellularly. In some embodiments, the target-binding moiety Y has approximately the same binding affinity for FcRn as wild-type IgG at pH 7.4.

[0525] In some embodiments, Y is a mutant form of the antibody, wherein Y has a YTE mutation.

[0526] In one method, a method for degrading a target molecule in a subject in need thereof is provided, the method comprising: a means for binding to a lysosomal targeting molecule extracellularly; a means for binding to a target molecule extracellularly; administering an effective amount of a conjugate comprising a means for dissociating from the target molecule in an endosome; The conjugate is externalized from the cell.

[0527] In some embodiments, the means for binding to the lysosome targeting molecule remains bound to the lysosome targeting molecule in the endosome. In one embodiment, the means for binding to the targeting molecule also binds to FcRn in the endosome. In one embodiment, the conjugate dissociates from the lysosome targeting molecule in the endosome.

[0528] 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, M6PR, FR). In some embodiments, one Y biomolecule is conjugated to one (Xn-L)-group; 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, tetravalent, etc.). In some embodiments of Formula (I), when Y is a biomolecule, it is understood that Y can be conjugated to more than one (Xn-L)-group, and each (Xn-L)-group itself may be monovalent or multivalent (e.g., divalent, trivalent, tetravalent, etc.). In such cases, the ratio of linked (Xn-L)-groups to biomolecules is said to be 2 or greater.

[0529] In some embodiments, one or more (Xn-L)- groups (e.g., m is 1, 2, or 3) are conjugated to Y to generate a residue moiety resulting from the covalent bonding of the chemoselective ligation group to a compatible group on Y. For example, conjugates of the present disclosure can be prepared using the building blocks described herein, as illustrated in Scheme 1. In Scheme 1, a conjugate of formula (I') [ka] Formula (II') [ka] wherein: n is 1 to 3, m is 1 to 20; a, b, c, d, and e are each independently 1, 2, 3, 4, or 5; L 1 ~L 6 are each independently a linking moiety, and together they form Z 1 and Y, forming a linear or branched linker therebetween; X and Y are as defined herein.

[0530] Scheme I is for illustrative purposes and is not intended to limit the scope of the disclosure in any way. However, as will be appreciated by those of skill in the art, compounds of the present disclosure have a variety of L moieties, which may be selected from the group consisting of X, -L ... 1 -part) to Z 1 and provides an exemplary target binding moiety, or component of X. In Scheme I, R M1 and R M2 are each independently a reactive functional group for coupling reactions (e.g., alkyne, -N, -C(O)OH, -NH, etc.), and Y' is a chemoselective ligation group that can be conjugated to the amino acid residue(s) of Y. [ka]

[0531] Methods and exemplary reagents and starting materials for each step (ie, compounds of Formulas 1-1, 1-2, 1-3) are described throughout or can be derived from the art.

[0532] In some embodiments, Y is an antibody or antibody fragment that specifically binds to a target protein, and the compound has formula (III): [ka] is a conjugate of n is 1 to 20; m is the average load from 1 to 80; each X is a ligand moiety that binds to a lysosomal targeting molecule; each L is a linker; each Z is a residue moiety resulting from the covalent bond between the chemoselective ligation group and a compatible group on the Ab; An Ab is an antibody or antibody fragment that specifically binds to a target protein.

[0533] In certain embodiments of the conjugate of Formula (III), L is a linker of Formula (II) (e.g., as described herein).

[0534] In certain embodiments of the conjugate of Formula (III), n is 1 to 6. In certain cases, n is 1, such that the antibody is conjugated to a monovalent ligand and the linker is of Formula (IIa) (e.g., as described herein). In certain cases, n is at least 2, such that the antibody is conjugated to a multivalent ligand. In certain cases, n is 2. In certain cases, n is 3.

[0535] In certain embodiments of the conjugate of Formula (III), Z is a residue moiety resulting from the covalent attachment of a chemoselective ligation moiety (eg, Tables 10 and 11).

[0536] In certain embodiments of the conjugate of formula (III), Z is a residue moiety resulting from the covalent bond between a thiol-reactive chemoselective ligation group and one or more cysteine ​​residue(s) of the Ab.

[0537] In certain other embodiments of the conjugate of Formula (III), Z is a residue moiety resulting from the covalent bond between the amine-reactive chemoselective ligation group and one or more lysine residue(s) of the Ab.

[0538] In certain embodiments, conjugates having the linker structures described herein have weak binding affinity to cell surface receptors. Without being bound by any particular mechanism or theory, such weaker binding affinity may be compensated for to increase the 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, such weaker binding conjugates are still capable of sufficiently strong uptake.

[0539] 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 can be prepared using any suitable method disclosed in the art (see, e.g., Bioconjugate Techniques (Hermanson ed., 2d ed. 2008)).

[0540] 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 from open cysteine-cysteine ​​disulfide bonds of the Ab. In certain embodiments, the open cysteine-cysteine ​​disulfide bonds are interchain disulfide bonds.

[0541] 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.

[0542] In certain embodiments, conjugation to a polypeptide or antibody Ab can be performed via site-specific conjugation. For example, site-specific conjugation can achieve uniform loading and minimize conjugate subpopulations that may alter antigen binding or pharmacokinetics. In certain embodiments, for example, conjugation can involve engineering cysteine ​​substitutions at positions on the polypeptide or antibody, such as on the heavy and / or light chains of the antibody, that provide reactive thiol groups, do not interfere with folding and assembly of the polypeptide or antibody, and do not alter polypeptide or antigen binding (see, e.g., Junutula et al., J. Immunol. Meth. 2008;332:41-52; and Junutula et al., Nature Biotechnol. 2008;26:925-32; see also WO2006 / 034488, the entire contents of which are incorporated herein by reference). In another non-limiting approach, selenocysteine ​​is co-translationally inserted into a polypeptide or antibody sequence by recoding the stop codon UGA from the terminus to insert a selenocysteine, allowing site-specific covalent attachment at 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 engineering unnatural amino acids, including, for example, 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, including, 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 entire contents of each of which are incorporated by reference. See also US 2019 / 0060481 A1 and US 2016 / 0060354 A1, the entire contents of each of which are incorporated by reference. All of these methodologies are contemplated for use in connection with making the conjugates described herein.

[0543] The loading 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 of a conjugate. The number of "X" moieties (e.g., folate moieties) per "Xn-L-" or "Xn-" unit is represented by "n" in Formula (III). As used herein, the term "valency" refers to the number of "X" moieties ("n") per unit. It will be understood that the loading, i.e., DAR, is not necessarily the same as 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, there will be 1x1=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 will be 2 x 4 = 8 "X" moieties per conjugate. Thus, for the conjugates described herein, the total number of "X" groups per conjugate molecule is n x m. As used herein, the term "total valency" refers to the total number of "X" moieties (n x m, total valency) per conjugate molecule.

[0544] The DAR(loading) can range from 1 to 80 units per conjugate. The conjugates provided herein can include a population of polypeptides, antibodies, or antigen-binding fragments conjugated with, for example, units ranging from 1 to 80. The average number of units per polypeptide or antibody in a preparation of conjugates resulting from a conjugation reaction can be characterized by conventional means, such as mass spectrometry. The quantitative distribution of DAR(loading) in terms of m can also be determined. In some cases, isolation, purification, and characterization of homogeneous conjugates with a particular value of m can be achieved by means such as electrophoresis.

[0545] In certain embodiments, the DAR for the conjugates provided herein ranges from 1 to 80. In certain embodiments, the DAR for the conjugates provided herein ranges from 1 to 70. In certain embodiments, the DAR for the conjugates provided herein ranges from 1 to 60. In certain embodiments, the DAR for the conjugates provided herein ranges from 1 to 50. In certain embodiments, the DAR for the conjugates provided herein ranges from 1 to 40. In certain embodiments, the DAR for the conjugates provided herein ranges from 1 to 35. In certain embodiments, the DAR for the conjugates provided herein ranges from 1 to 30. In certain embodiments, the DAR for the conjugates provided herein ranges from 1 to 25. In certain embodiments, the DAR for the conjugates provided herein ranges from 1 to 20. In certain embodiments, the DAR for the conjugates provided herein ranges from 1 to 18. In certain embodiments, the DAR for the conjugates provided herein ranges from 1 to 15. In certain embodiments, the DAR for the conjugates provided herein ranges from 1 to 12. In certain embodiments, the DAR for the conjugates provided herein ranges from 1 to 10. In certain embodiments, the DAR for the conjugates provided herein ranges from 1 to 9. In certain embodiments, the DAR for the conjugates provided herein ranges from 1 to 8. In certain embodiments, the DAR for the conjugates provided herein ranges from 1 to 7. In certain embodiments, the DAR for the conjugates provided herein ranges from 1 to 6. In certain embodiments, the DAR for the conjugates provided herein ranges from 1 to 5. In certain embodiments, the DAR for the conjugates provided herein ranges from 1 to 4. In certain embodiments, the DAR for the conjugates provided herein ranges from 1 to 3.In certain embodiments, the DAR for the conjugates provided herein ranges from 2 to 12. In certain embodiments, the DAR for the conjugates provided herein ranges from 2 to 10. In certain embodiments, the DAR for the conjugates provided herein ranges from 2 to 9. In certain embodiments, the DAR for the conjugates provided herein ranges from 2 to 8. In certain embodiments, the DAR for the conjugates provided herein ranges from 2 to 7. In certain embodiments, the DAR for the conjugates provided herein ranges from 2 to 6. In certain embodiments, the DAR for the conjugates provided herein ranges from 2 to 5. In certain embodiments, the DAR for the conjugates provided herein ranges from 2 to 4. In certain embodiments, the DAR for the conjugates provided herein ranges from 3 to 12. In certain embodiments, the DAR for the conjugates provided herein ranges from 3 to 10. In certain embodiments, the DAR for the conjugates provided herein ranges from 3 to 9. In certain embodiments, the DAR range for the conjugates provided herein is 3 to 8. In certain embodiments, the DAR range for the conjugates provided herein is 3 to 7. In certain embodiments, the DAR range for the conjugates provided herein is 3 to 6. In certain embodiments, the DAR range for the conjugates provided herein is 3 to 5. In certain embodiments, the DAR range for the conjugates provided herein is 3 to 4.

[0546] In certain embodiments, the DAR for 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 for 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. In some embodiments, the DAR for the conjugates provided herein is about 2.2.

[0547] In some embodiments, the DAR range for the conjugates provided herein is 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 range for the conjugates provided herein is 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 for the conjugates provided herein is about 1. In some embodiments, the DAR for the conjugates provided herein is about 2. In some embodiments, the DAR for the conjugates provided herein is about 3. In some embodiments, the DAR for the conjugates provided herein is about 4. In some embodiments, the DAR for the conjugates provided herein is about 3.8. In some embodiments, the DAR for the conjugates provided herein is about 5. In some embodiments, the DAR for the conjugates provided herein is about 6. In some embodiments, the DAR for a conjugate provided herein is about 7. In some embodiments, the DAR for a conjugate provided herein is about 8. In some embodiments, the DAR for a conjugate provided herein is about 9. In some embodiments, the DAR for a conjugate provided herein is about 10. In some embodiments, the DAR for a conjugate provided herein is about 11. In some embodiments, the DAR for a conjugate provided herein is about 12. In some embodiments, the DAR for a conjugate provided herein is about 13. In some embodiments, the DAR for a conjugate provided herein is about 14. In some embodiments, the DAR for a conjugate provided herein is about 15. In some embodiments, the DAR for a conjugate provided herein is about 16. In some embodiments, the DAR for a conjugate provided herein is about 17.In some embodiments, the DAR for the conjugates provided herein is about 18. In some embodiments, the DAR for the conjugates provided herein is about 19. In some embodiments, the DAR for the conjugates provided herein is about 20.

[0548] In some embodiments, the DAR for a conjugate provided herein is about 25. In some embodiments, the DAR for a conjugate provided herein is about 30. In some embodiments, the DAR for a conjugate provided herein is about 35. In some embodiments, the DAR for a conjugate provided herein is about 40. In some embodiments, the DAR for a conjugate provided herein is about 50. In some embodiments, the DAR for a conjugate provided herein is about 60. In some embodiments, the DAR for a conjugate provided herein is about 70. In some embodiments, the DAR for a conjugate provided herein is about 80.

[0549] In certain embodiments, fewer than the theoretical maximum number of units are conjugated to a polypeptide, e.g., an antibody, during the conjugation reaction. The polypeptide may contain, for example, lysine residues that do not react with a compound or linker reagent. Generally, for example, antibodies do not contain many free reactive cysteine ​​thiol groups that can be linked to a drug unit; in fact, most cysteine ​​thiol residues in antibodies exist as disulfide bonds. In certain embodiments, antibodies can be reduced with a reducing agent such as dithiothreitol (DTT) or tricarbonylethylphosphine (TCEP) under partial or complete reducing conditions to generate reactive cysteine ​​thiol groups. In certain embodiments, the antibody is subjected to denaturing conditions to expose reactive nucleophilic groups, such as lysine or cysteine. In some embodiments, the 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.

[0550] In certain embodiments, the amino acid attached to the unit is in the heavy chain of the antibody. In certain embodiments, the amino acid attached to the unit is in the light chain of the antibody. In certain embodiments, the amino acid attached to the unit is in the hinge region of the antibody. In certain embodiments, the amino acid attached to the unit is in the Fc region of the antibody. In certain embodiments, the amino acid attached to the unit is in a constant region of the antibody (e.g., CH1, CH2, or CH3 of the heavy chain, or CH1 of the light chain). In still other embodiments, the amino acid attached to the unit or Drug Unit is in a VH framework region of the antibody. In still other embodiments, the amino acid attached to the unit is in a VL framework region of the antibody.

[0551] The DAR (loading) of the conjugate can be controlled in various ways, such as (i) limiting the molar excess of compound or coupling reagent relative to the polypeptide, (ii) limiting the coupling reaction time or temperature, (iii) partial or limiting reducing conditions for cysteine ​​thiol modification, or (iv) recombinantly modifying the amino acid sequence of the polypeptide to alter the number and position of cysteine ​​residues to control the number and / or position of linker-drug attachments (e.g., in the case of thiomab prepared as disclosed in WO2006 / 034488, which is incorporated herein by reference in its entirety).

[0552] It should be understood that preparation of the conjugates described herein may result in a mixture of multiple conjugates having a distribution of one or more units attached to a polypeptide, e.g., an antibody. Individual conjugate molecules in the mixture can be identified by mass spectrometry and separated by HPLC, e.g., hydrophobic interaction chromatography, including methods known in the art. In certain embodiments, homogeneous conjugates with a single DAR (loading) value can be separated from the conjugation mixture by electrophoresis or chromatography.

[0553] 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. In certain cases, m is 10 or less. In certain cases, m is 2 to 8. In certain cases, m is 2 to 6. In certain cases, m has an average weight of about 4.

[0554] It should be understood that preparation of the conjugates described herein may result in a mixture of multiple conjugates having a distribution of one or more units attached to a polypeptide, e.g., an antibody. Individual conjugate molecules in the mixture can be identified by mass spectrometry and separated by HPLC, e.g., hydrophobic interaction chromatography, including methods known in the art. In certain embodiments, homogeneous conjugates with a single DAR (loading) value can be separated from the conjugation mixture by electrophoresis or chromatography.

[0555] antibody In some embodiments, the target binding moiety Y is an antibody or antibody fragment that specifically binds to a target molecule, such as a target protein.

[0556] The ligand moiety can be site-specifically covalently bound to the antibody or antibody fragment via any linker. The ligand moiety can be covalently bound to the antibody or antibody fragment via a site-specific cysteine ​​modification on the antibody or antibody fragment (e.g., L443C) and a thiol-reactive chemoselective ligation group. The lysosomal molecule-binding moiety can be covalently bound to the antibody or antibody fragment via one or more lysine residues on the antibody or antibody fragment and an amine-reactive chemoselective ligation group.

[0557] In some embodiments, the target-binding moiety is an antibody, and the ligand moiety is conjugated to the antibody via a linker using a thiol-reactive conjugate at L443C. In some embodiments, the target-binding moiety is a mutant of omalizumab, and the linker is conjugated to L443C. In some embodiments, the target-binding moiety is a mutant of ligelizumab, and the linker is conjugated to L443C. Other sites that support thiol-reactive conjugation include heavy chain A118C, A140C, K392C, K290C, S293C, and light chain K183C, V205C, and K149C. Other sites suitable for conjugation can also be used, for example, as reported in R. Ohri et al., Bioconjugate Chem. 2018, 19, 473-85.

[0558] In some embodiments, the conjugates of the present disclosure comprise an antibody (Ab), i.e., the target-binding moiety Y is 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.

[0559] In some embodiments, the antibody or antibody fragment comprises an Fc region, hi some embodiments, the antibody or antibody fragment comprises an Fc region comprising one or more mutations that increase the binding affinity of the antibody or antibody fragment to FcRn.

[0560] In some embodiments, the antibody or antibody fragment comprises an Fc region with one or more mutations selected from methionine (Met) to tyrosine (Tyr), serine (Ser) to threonine (Thr), and threonine (Thr) to glutamic acid (Glu) amino acid substitutions. For example, the antibody or antibody fragment can comprise one or more of M252Y, S254T, and T256E (commonly referred to as YTE mutations).

[0561] In some embodiments, the antibody or antibody fragment has pH-dependent binding affinity for the target molecule. In some embodiments, the antibody or antibody fragment has higher binding affinity for the target molecule at neutral pH than at low pH. In some embodiments, the antibody or antibody fragment has calcium-dependent binding affinity for the target molecule. In some embodiments, the antibody or antibody fragment has one or more mutations that confer pH-dependent binding affinity for the target molecule. In some embodiments, the target-binding moiety Y is an antibody or antibody fragment mutated from wild-type with one or more histidine substitutions. In some embodiments, the one or more histidine substitutions are located in the CDR regions of the antibody or antibody fragment. For example, certain antibodies or fragments thereof can include one or more mutations in the table below. [Table 26]

[0562] In the examples described herein, the numbering of Omalizumab follows a simple sequence numbering scheme, i.e., for Oma S35H-LC, Y57H-LC, residue 35 corresponds to Kabat number 31 and residue 57 corresponds to Kabat number 53. The numbering of Ligelizumab (e.g., Lige W33H-HC / Y50H-LC / N100bH-HC / W94H-LC) follows the Kabat numbering scheme.

[0563] In some embodiments, the target-binding moiety Y has a higher binding affinity for the target molecule outside the cell than within the endosome. In some embodiments, the target-binding moiety Y has a higher binding affinity for the target molecule at extracellular pH than at intraendosomal pH. In some embodiments, the target-binding moiety Y has a higher binding affinity for the target molecule at a pH of about 6.5 to about 7.5 than at a pH of about 6.5 to 4.5. In some embodiments, the target-binding moiety Y has a higher binding affinity for the target molecule at a pH of about 7.0 to about 7.5 than at a pH of about 6.5 to 4.5. In some embodiments, the target-binding moiety Y has a higher binding affinity for the target molecule at a pH of about 7.4 than at a pH of about 6.0. In some embodiments, the target-binding moiety Y binds to intraendosomal Ca 2+ Extracellular Ca concentration 2+ In some embodiments, the target binding moiety Y has a high binding affinity to the target molecule at Ca 2+ than when the concentration is about 0-1 μM. 2+ It has high binding affinity to the target molecule when its concentration is about 1 to 2 mM.

[0564] In some embodiments, the target binding moiety Y has a higher K for the target molecule in the endosome than outside the cell. D In some embodiments, the target binding moiety Y has an endosomal:extracellular K of 2:1 to 10,000:1 relative to the target molecule. D In some embodiments, the target binding moiety Y has an intraendosomal:extracellular K ratio of 2:1 to 1,000:1 relative to the target molecule. D In some embodiments, the target binding moiety Y has an intraendosomal:extracellular K ratio of 2:1 to 500:1 relative to the target molecule. D In some embodiments, the target binding moiety Y has an intraendosomal:extracellular K ratio of 2:1 to 100:1 relative to the target molecule. D In some embodiments, the target binding moiety Y has an intraendosomal:extracellular K ratio of 2:1 to 50:1 relative to the target molecule. DIn some embodiments, the target binding moiety Y has an intraendosomal:extracellular K ratio of 2:1 to 40:1 relative to the target molecule. D In some embodiments, the target binding moiety Y has an intraendosomal:extracellular K ratio of 2:1 to 30:1 relative to the target molecule. D In some embodiments, the target binding moiety Y has an intraendosomal:extracellular K ratio of 2:1 to 20:1 relative to the target molecule. D In some embodiments, the target binding moiety Y has an intraendosomal:extracellular K ratio of 10:1 to 1,000:1 relative to the target molecule. D In some embodiments, the target binding moiety Y has an intraendosomal:extracellular K ratio of 10:1 to 100:1 relative to the target molecule. D In some embodiments, the target binding moiety Y has a K ratio of 2:1 to 10,000:1 at pH 6.0:7.4 relative to the target molecule. D In some embodiments, the target binding moiety Y has a K ratio of 2:1 to 1,000:1 at pH 6.0:7.4 relative to the target molecule. D In some embodiments, the target binding moiety Y has a K ratio of 2:1 to 500:1 at pH 6.0:7.4 relative to the target molecule. D In some embodiments, the target binding moiety Y has a K ratio of 2:1 to 100:1 relative to the target molecule at pH 6.0:7.4. D In some embodiments, the target binding moiety Y has a K ratio of 2:1 to 50:1 relative to the target molecule at pH 6.0:7.4. D In some embodiments, the target binding moiety Y has a K ratio of 2:1 to 40:1 relative to the target molecule at pH 6.0:7.4. D In some embodiments, the target binding moiety Y has a K ratio of 2:1 to 30:1 relative to the target molecule at pH 6.0:7.4. D In some embodiments, the target binding moiety Y has a K ratio of 2:1 to 20:1 relative to the target molecule at pH 6.0:7.4. D In some embodiments, the target binding moiety Y has a K ratio of 10:1 to 1,000:1 at pH 6.0:7.4 relative to the target molecule. DIn some embodiments, the target binding moiety Y has a K ratio of 10:1 to 100:1 at pH 6.0:7.4 relative to the target molecule. D It has a ratio.

[0565] In some embodiments, the target binding moiety Y has a higher k for the target molecule in the endosome than outside the cell. off In some embodiments, the target binding moiety Y has a higher k for the target molecule at endosomal pH than at extracellular pH. off In some embodiments, the target binding moiety Y has a higher k for the target molecule at pH 6.0 than the corresponding wild type. off In some embodiments, the target binding moiety Y is mutated to have a binding efficiency of 10 to the target molecule in the endosome. -4 ~10 -1 (1 / s) k off In some embodiments, the target binding moiety Y has a binding ratio of 10 to the target molecule in the endosome. -3 ~10 -1 (1 / s) k off In some embodiments, the target binding moiety Y has a binding ratio of 10 to the target molecule in the endosome. -2 ~10 -1 (1 / s) k off has a rate.

[0566] In some embodiments, the antibody or antibody fragment specifically binds to a cancer antigen.

[0567] In some embodiments, the antibody or antibody fragment specifically binds to a hepatocyte antigen.

[0568] In some embodiments, the antibody or antibody fragment specifically binds to an antigen displayed on a macrophage.

[0569] In some embodiments, the antibody or antibody fragment specifically binds to the full complement or a fragment thereof, hi some embodiments, the antibody or antibody fragment specifically binds to one or more immunodominant epitope(s) within the full complement or a fragment thereof.

[0570] 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.

[0571] 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.

[0572] 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.

[0573] In some embodiments, the antibody or antibody fragment specifically binds to a vascular endothelial growth factor (VEGF) protein, e.g., a human VEGF protein. In some embodiments, the antibody or antibody fragment specifically binds to one or more immunodominant epitopes within the VEGF protein.

[0574] 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.

[0575] In some embodiments, the antibody or antibody fragment specifically binds to a fibroblast growth factor (FGF), such as human FGF, hi some embodiments, the antibody or antibody fragment specifically binds to one or more immunodominant epitopes within the FGF protein.

[0576] In some embodiments, the antibody or antibody fragment specifically binds to a fibroblast growth factor receptor (FGFR), e.g., human FGFR. In some embodiments, the antibody or antibody fragment specifically binds to a fibroblast growth factor receptor 2 (FGFR2) protein, e.g., human FGFR2 protein, e.g., FGFR2b protein. In some embodiments, the antibody or antibody fragment specifically binds to a fibroblast growth factor receptor 3 (FGFR3) protein, e.g., 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.

[0577] 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.

[0578] 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.

[0579] In some embodiments, the antibody specifically binds to an immune checkpoint inhibitor. In some embodiments, the antibody binds to one or more immunodominant epitope(s) 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 epitope(s) within the PD-1 protein.

[0580] In some embodiments, the antibody specifically binds to a programmed death-ligand 1 (PD-L1) protein, e.g., human PD-L1, hi some embodiments, the antibody specifically binds to one or more immunodominant epitope(s) within the PD-L1 protein.

[0581] In some embodiments, the antibody binds to TIM3, hi some embodiments, the antibody binds to one or more immunodominant epitope(s) within TIM3.

[0582] In some embodiments, the antibody specifically binds to a lectin. In some embodiments, the antibody specifically binds to one or more immunodominant epitope(s) within the lectin. In some embodiments, the antibody binds to SIGLEC. In some embodiments, the antibody binds to one or more immunodominant epitope(s) within SIGLEC. In some embodiments, the antibody binds to a cytokine receptor. In some embodiments, the antibody binds to one or more immunodominant epitope(s) within a cytokine receptor. In some embodiments, the antibody binds to sIL6R. In some embodiments, the antibody binds to one or more immunodominant epitope(s) within sIL6R. In some embodiments, the antibody binds to a cytokine. In some embodiments, the antibody binds to one or more immunodominant epitope(s) 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.

[0583] 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 beta2 microglobulin. In some embodiments, the antibody binds to one or more immunodominant epitope(s) within beta2 microglobulin. In some embodiments, the antibody binds to IgE. In some embodiments, the antibody is omalizumab. In some embodiments, the antibody is a mutant form of omalizumab. In some embodiments, the antibody is ligelizumab. In some embodiments, the antibody is a mutant form of ligelizumab. In some embodiments, the antibody binds to hC5. In some embodiments, the antibody is eculizumab or a mutant thereof. In some embodiments, the antibody is ALXN1210 or a mutant thereof.

[0584] In some embodiments, the antibody is omalizumab, rigelzimab, eculizumab, ALXN1210, or a mutant thereof. Mutant means that the antibody retains at least about 90%, 95%, or 97% of the functionality or binding affinity for its intended target compared to the wild-type antibody, but includes one or more mutations described herein.

[0585] In some embodiments, the antibody is omalizumab or a mutant thereof. The sequence of wild-type omalizumab is publicly known. For example, the DrugBank Accession No. for omalizumab is DB00043 (go.drugbank.com / drugs / DB00043). In some embodiments, omalizumab contains one or more of the following mutations: S35H(LC), Y57H(LC), L443C(HC), M252Y(HC), S254T(HC), and T256E(HC). In some embodiments, omalizumab contains the following mutations: S35H(LC), Y57H(LC), L443C(HC), M252Y(HC), S254T(HC), and T256E(HC).

[0586] In some embodiments, the antibody is ligelizumab or a mutant thereof. The sequence of wild-type ligelizumab is publicly known. For example, the KEGG Entry ID for ligelizumab is D11761 (www.kegg.jp / entry / D11761). In some embodiments, ligelizumab contains one or more of the following mutations: W33H(HC), Y50H(LC), N100bH(HC), W94H(LC), L443C(HC), M252Y(HC), S254T(HC), and T256E(HC). In some embodiments, ligelizumab contains the following mutations: W33H(HC), Y50H(LC), N100bH(HC), W94H(LC), L443C(HC), M252Y(HC), S254T(HC), and T256E(HC). W33H(HC), Y50H(LC), N100bH(HC), W94H(LC), L443C(HC), M252Y(HC), S254T(HC), and T256E(HC).

[0587] In some embodiments, the antibody is eculizumab or a derivative thereof. The sequence of wild-type eculizumab is publicly known. For example, the NCATS No. of eculizumab is A3ULP0F556 (drugs.ncats.io / substance / A3ULP0F556). In some embodiments, eculizumab contains one or more of the following mutations: L443C(HC), M252Y(HC), S254T(HC), and T256E(HC). In some embodiments, eculizumab contains the following mutations: L443C(HC), M252Y(HC), S254T(HC), and T256E(HC).

[0588] In some embodiments, the antibody is ALXN1210 or a derivative thereof. The sequence of wild-type ALXN1210 is publicly known. For example, the DrugBank Accession No. for ALXN1210 is DB11580 (go.drugbank.com / drugs / DB11580). In some embodiments, ALXN1210 contains one or more of the following mutations: L443C(HC), M252Y(HC), S254T(HC), and T256E(HC). In some embodiments, ALXN1210 contains the following mutations: L443C(HC), M252Y(HC), S254T(HC), and T256E(HC).

[0589] target As summarized above, the bifunctional compounds of the present disclosure can include an antibody (Y) specifically bound to a target molecule, which can be a cell surface molecule or an extracellular molecule.

[0590] In some embodiments of the compounds and methods of the present disclosure, the target molecule is a cell surface molecule. "Cell surface molecule" refers to a target molecule associated with a cell membrane, for example, 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 can be any cell surface molecule for which targeted degradation via the endosomal / lysosomal pathway is desired. In some embodiments, the cell surface molecule is a cell surface receptor.

[0591] 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.

[0592] In some embodiments, antibody (Y) specifically binds to a cell surface molecule, and its effect is mediated through a bulk biophysical or collective effect rather than through a specific molecular interaction (and therefore less susceptible to inhibition). 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.

[0593] In some embodiments, when an antibody 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 that can be characterized by one or more of aberrant cell growth, aberrant cell 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 suitable indicators of cellular transformation. As used herein, "cancer cell" is used interchangeably with "tumor cell," "malignant cell," or "cancerous cell" and encompasses cancer cells such as solid tumors, semi-solid tumors, hematopoietic malignancies (e.g., leukemia cells, lymphoma cells, myeloma cells, etc.), primary tumors, and metastatic tumors. In some embodiments, the cell surface molecule present on a cancer cell is a tumor-associated antigen or a tumor-specific antigen. In certain aspects, when an antibody (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 an 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 can 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 can be inhibited according to the methods of the present disclosure include C-type lectins, including, but not limited to, CLEC4A (DCIR), Ly49Q, and MICL.Further details regarding inhibitory immunoreceptors are described, for example, in Steevels et al. (2011) Eur. J. Immunol. 41(3):575-587. In some embodiments, the cell surface molecule present on an immune cell is a ligand of an inhibitory immunoreceptor. In certain aspects, the cell surface molecule present on an immune cell is an immune checkpoint molecule. Non-limiting examples of immune checkpoint molecules to which the moiety of interest (Y) can specifically bind include PD-1, PD-L1, CTLA4, TIM3, LAG3, TIGIT, and members of the B7 family.

[0594] In some embodiments of the compounds and methods of the present disclosure, the target molecule is an extracellular molecule. "Extracellular molecule" refers to a soluble molecule that is located outside the cell membrane of any cell in the vicinity of a soluble molecule. The extracellular molecule can be any extracellular molecule for which targeted degradation via the endosomal / lysosomal pathway is desired.

[0595] 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), a chemokine or cytokine (e.g., a mediator of sepsis or chronic inflammation such as IL-1), a protein hormone (e.g., insulin, ACTH, etc.), a protein mediator of mood disorders, 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., in the case of peanut allergy), a protein toxin (e.g., snake venom hyaluronidase, etc.), an autoantibody, etc.

[0596] In some embodiments, the target molecule is an extracellular molecule that is an antibody, e.g., an antibody specifically bound 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, 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, which is expressed on immune effector cells, initiating an inflammatory response. Abnormal IgA expression is thought to be associated with 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 be attached 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 IgG4-associated diseases. In some embodiments, the target is human immunoglobulin E (IgE). IgE is a type of immunoglobulin that plays a key role in type I hypersensitivity, which can lead to a variety of allergic diseases and conditions.

[0597] 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 β (TGF-β), including any specific subtypes of such cytokines), hormones, etc. In certain aspects, antibody (Y) specifically binds to IgE.

[0598] Pharmaceutical Composition In another embodiment, provided herein is a pharmaceutical composition comprising one or more conjugates disclosed herein and a pharmaceutically acceptable carrier.

[0599] In certain embodiments, the pharmaceutical compositions provided herein contain a therapeutically effective amount of one or more conjugates provided herein, and optionally one or more additional prophylactic or therapeutic agents, in a pharmaceutically acceptable carrier.

[0600] Pharmaceutical carriers suitable for administration of the conjugates provided herein include any carriers known to those skilled in the art to be suitable for the particular method of administration.

[0601] The conjugates described herein can be formulated as the sole pharmaceutically active ingredient in the composition or can be combined with other active ingredients.

[0602] 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 transdermal patch formulations and dry powder inhalants.

[0603] In the compositions provided herein, the conjugates described herein can be mixed with a suitable pharmaceutical carrier. The concentration of the conjugate in the composition can be, for example, effective to deliver an amount that, upon administration, treats, prevents, or ameliorates a condition or disorder described herein or a symptom thereof.

[0604] In certain embodiments, the pharmaceutical compositions provided herein are formulated for single-dose administration. To formulate the composition, the weight fraction of the conjugate is dissolved, suspended, dispersed, or otherwise mixed in the selected carrier at an effective concentration such that the symptoms to be treated are alleviated, prevented, or one or more symptoms are ameliorated.

[0605] The concentration of the conjugate in the pharmaceutical compositions provided herein depends, for example, on the physicochemical properties of the conjugate, the administration schedule, the dosage, as well as other factors known to those of skill in the art.

[0606] 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 an appropriate 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, such as 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 forms or multi-dose forms. As used herein, unit dosage form refers to a physically discrete unit suitable for human or animal (e.g., mammalian) subjects and individually packaged 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, syringes, and individually packaged capsules. The unit dosage form can be administered in divided doses or multiple times. A multiple-dosage form is a dosage form in which a plurality of identical unit-dosage forms are packaged in one container and administered in separate unit-dosage forms.Examples of multiple-dosage forms include vials, capsule bottles, or bottles.Therefore, in certain embodiments, a multiple-dosage form is a multiple of the unit-dosage form that is not separated in packaging.

[0607] In certain embodiments, the conjugates herein are in the form of liquid pharmaceutical formulations. Liquid pharmaceutical formulations can be prepared, for example, by dissolving, dispersing, or mixing the conjugates and optional auxiliary agents 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 can also contain small amounts of non-toxic auxiliary substances such as wetting agents, emulsifiers, solubilizers, pH buffering agents, etc.

[0608] 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 can be prepared containing antibodies in the range of 0.005% to 100%, with the remainder consisting of non-toxic carriers.

[0609] In certain embodiments, parenteral administration is characterized by subcutaneous, intramuscular, or intravenous injection, although parenteral administration is also contemplated herein. Injectables can be prepared in conventional forms, such as liquid solutions or suspensions, solid forms suitable for dissolving or suspending in liquid before injection, or emulsions. Injectables, solutions, and emulsions also contain one or more excipients. Suitable excipients are, for example, water, saline, glucose, glycerol, or ethanol. Other administration routes may include enteral administration, intracerebral administration, nasal administration, intraarterial administration, intracardiac administration, intraosseous injection, intraspinal administration, and intraperitoneal administration.

[0610] Preparations for parenteral administration include sterile solutions for injection, sterile dry soluble products (including subcutaneous tablets) such as lyophilized powders to be combined with a solvent immediately before use, sterile suspensions for injection, sterile dry insoluble products to be combined with a vehicle immediately before use, and sterile emulsions. The solutions may be aqueous or non-aqueous.

[0611] For intravenous administration, suitable carriers include saline or phosphate buffered saline (PBS), as well as solutions containing thickening and solubilizing agents, such as glucose, polyethylene glycol, polypropylene glycol, and mixtures thereof.

[0612] 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.

[0613] Pharmaceutical carriers also include ethyl alcohol, polyethylene glycol, propylene glycol for water miscible vehicles and sodium hydroxide, hydrochloric acid, citric acid or lactic acid for pH adjustment.

[0614] In certain embodiments, intravenous or intraarterial infusion of a sterile aqueous solution containing a conjugate described herein is an effective method 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.

[0615] In certain embodiments, the pharmaceutical formulation is a lyophilized powder that can be reconstituted for administration as a solution, emulsion, or other mixture, or can be formulated as a solid or gel upon reconstitution.

[0616] The lyophilized powder is prepared by dissolving the conjugate described herein in a suitable solvent. In some embodiments, the lyophilized powder is sterilized. The suitable solvent may contain excipients or other pharmacological ingredients that improve the stability of the powder or a reconstitution solution prepared from the powder. Excipients that can be used include, but are not limited to, dextrose, sorbitol, fructose, corn syrup, xylitol, glycerin, glucose, sucrose, or other suitable agents. The suitable solvent may also contain a buffer solution such as citrate, sodium phosphate, potassium phosphate, or other buffers known to those of skill in the art (in certain embodiments, 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 divided into vials for lyophilization. The lyophilized powder can be stored under appropriate conditions, such as at about 4°C to room temperature.

[0617] This lyophilized powder is reconstituted with water for injection to provide a formulation for parenteral administration. To reconstitute, the lyophilized powder is added to sterile water or other suitable carrier.

[0618] In certain embodiments, the conjugates provided herein can be formulated for topical administration or topical application, for example, in the form of gel, cream, and lotion, for topical application to the skin and mucous membranes, such as the eye, and for application to the eye, or for intravesical or intrathecal application.Topical administration is considered for transdermal delivery, administration to the eye or mucous membrane, or for inhalation therapy.Nasal drops of the active compound alone or in combination with other pharmaceutically acceptable excipients can also be administered.

[0619] Uses 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. 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.

[0620] Removal of a target protein can refer to the reduction or depletion of the target protein from the cell surface or extracellular space or extracellular environment, i.e., the reduction or depletion of the amount of the target protein on the cell surface or in the extracellular environment.

[0621] In one aspect, provided herein is a method for sequestering a polypeptide of interest (target protein) in the lysosomes of a cell using a conjugate 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 a conjugate described herein and degrading the polypeptide of interest.

[0622] In one aspect, provided herein are methods for degrading a polypeptide of interest (target protein) using the conjugates described herein.

[0623] 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.

[0624] In another aspect, provided herein is a method for depleting a polypeptide of interest (target protein) described herein by administering to a subject in need of treatment an effective amount of a conjugate or pharmaceutically acceptable salt described herein, or a pharmaceutical composition described herein. In certain embodiments, the subject is a mammal (e.g., a human).

[0625] In one embodiment, a method for degrading a target molecule in a subject in need thereof is provided, comprising administering to the subject an effective amount of a conjugate described herein. In one embodiment, at least 90% of the target is degraded 4 days after administration. In one embodiment, at least 90% of the target is degraded 7 days after administration. In one embodiment, the superstoichiometric target:conjugate ratio is reduced. In one embodiment, the ratio is about 5. In one embodiment, the ratio is about 10. In one embodiment, the ratio is between 10 and 100. In one embodiment, the ratio is between 100 and 1000. In one embodiment, the target-binding moiety Y has a higher binding affinity for the target molecule extracellularly than in the endosome. In one embodiment, the target is IgE. In one embodiment, the conjugate promotes degradation of a molar excess of the target relative to the conjugate. In one embodiment, the molar excess is about 5. In one embodiment, the molar excess is about 10. In one embodiment, the molar excess is between 10 and 100. In one embodiment, the molar excess is about 100-1000.

[0626] definition It is to be understood that the present disclosure is not limited to particular embodiments described, which 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.

[0627] 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.

[0628] It should be noted that as used in this specification and the appended claims, the singular forms "a," "and," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to a "compound" includes not only a single compound but also a combination of two or more compounds, reference to a "substituent" includes not only a single substituent but also two or more substituents, etc.

[0629] In describing and claiming the present invention, certain terms will be used in accordance with the definitions set forth below. It will be understood that the definitions provided herein are not intended to be mutually exclusive. Accordingly, some chemical moieties may fall under more than one term definition.

[0630] As used herein, the words "for example," "for instance," "such as," or "including" are intended to introduce examples that further clarify a more general subject matter. These examples are provided solely as an aid in understanding the disclosure and are not intended to be limiting in any way.

[0631] 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 dates of publication provided may be different from the actual publication dates, which may need to be independently confirmed.

[0632] The terms "protein" and "polypeptide" are used interchangeably. Proteins may contain moieties other than amino acids (e.g., glycoproteins, etc.) and / or may be otherwise processed or modified. Those of skill in the art will understand that a "protein" can be 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 can include one or more protein chains attached non-covalently or covalently, e.g., linked by one or more disulfide bonds, or associated by other means. In certain embodiments, polypeptides can exist as a single chain or as two or more associated chains, and can exist as multimers, e.g., dimers, trimers, etc. These terms also include amino acid polymers that are modified naturally or by intervention (e.g., by disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or other manipulation or modification). The definition also includes 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, etc. 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.

[0633] The terms "antibody" and "immunoglobulin" are terms of art and may be used interchangeably herein in their broadest sense to include a specific type of immunoglobulin molecule that contains one or more antigen-binding domains that specifically bind to an antigen or epitope.

[0634] In certain embodiments, an isolated antibody (e.g., a monoclonal antibody), or antigen-binding fragment thereof, described herein specifically binds to a protein of interest, e.g., EGFR, and is conjugated to one or more lysosomal targeting moieties, e.g., via a linker.

[0635] An "antigen" is a moiety or molecule that contains an epitope to which an antibody can specifically bind. Thus, an antigen is also specifically bound by an antibody. 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).

[0636] The terms "clear," "degrade," "remove," and their synonyms refer to the degradation of a target or protein of interest, such as in a lysosome or late endosome. Synonyms such as "clear" or "clearance" can also refer to the removal of a target (or protein of interest) from the extracellular environment, such as serum or medium surrounding a cell.

[0637] "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 amino acids from two or more non-contiguous regions of the antigen.

[0638] The terms "bind," "binds," "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 also bind to other polypeptides, which will generally result in lower affinity as determined, 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 has a Kd At least 2 log, 2.5 log, 3 log, or 4 log lower (higher affinity) affinity (K d ) binds to the antigen. In another specific embodiment, a molecule that specifically binds to an antigen does not cross-react with other proteins. In another specific embodiment, when EGFR is the protein of interest, a molecule that specifically binds to an antigen does not cross-react with other non-EGFR proteins.

[0639] Antibodies include full-length antibodies (e.g., complete immunoglobulins), antibody fragments, monoclonal antibodies, polyclonal antibodies, recombinant antibodies, monospecific antibodies, multispecific antibodies (including bispecific antibodies), human antibodies, humanized antibodies, chimeric antibodies, synthetic antibodies, tetrameric antibodies comprising two heavy chain molecules 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, and antibodies having two light chain / heavy chain pairs (e.g., identical pairs). Antibodies include, but are not limited to, antibodies, 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.

[0640] An antibody can be any type (e.g., IgG, IgE, IgM, IgD, IgA, or IgY), any class (IgG1, IgG2, IgG3, IgG4, IgA1, or IgA2), or any subclass (IgG2a or IgG2b) of immunoglobulin molecule. In certain embodiments, the antibodies described herein are IgG antibodies (e.g., human IgG), or classes (e.g., human IgG1, IgG2, IgG3, or IgG4) or subclasses thereof.

[0641] In certain embodiments, the antibody is a four-chain antibody unit comprising two heavy (H) / 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 and L chains comprise constant regions, e.g., human constant regions. In even more specific 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.

[0642] The term "constant region" or "constant domain" is a well-known antibody terminology (sometimes referred to as "Fc") that 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 which may exert various effector functions, such as interacting with Fc receptors. These terms refer to portions of immunoglobulin molecules that generally have more conserved amino acid sequences than immunoglobulin variable domains.

[0643] The term "heavy chain" as used in reference to an antibody can refer to any of the different types, such as alpha (α), delta (δ), epsilon (ε), gamma (γ), mu (μ), etc., based on the amino acid sequence of the constant domain, which give rise to antibodies of the IgA, IgD, IgE, IgG, and IgM classes, respectively, and also include subclasses of IgG, such as IgG1, IgG2, IgG3, and IgG4.

[0644] The term "light chain" as 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 the constant domain. Light chain amino acid sequences are well known in the art. In certain embodiments, the light chain is a human light chain.

[0645] 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 a particular method for making the antibody. Generally, a population of monoclonal antibodies can be produced by a cell, a cell population, 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 host cell producing a recombinant antibody), and the antibody specifically binds to an epitope as determined, for example, by ELISA or other antigen-binding or competitive binding assays known in the art or shown in the Examples described 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 antibody or a multispecific antibody (e.g., a bispecific antibody).

[0646] The term "variable region" or "variable domain" refers to a portion of an antibody, generally a light or heavy chain, typically approximately 110-120 amino acids at the amino terminus of a mature heavy chain and approximately 90-100 amino acids at the amino terminus of a mature light chain. The variable region contains complementarity-determining regions (CDRs) flanked by framework regions (FRs). Generally, the spatial orientation of the CDRs and FRs, from N- to C-terminus, is as follows: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4. While not 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 antibody-antigen interactions and the specificity of the antibody for its epitope. In certain embodiments, the numbering of amino acid positions in the antibodies described herein follows the EU Index as set forth in Kabat et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242. In certain embodiments, the variable region is a human variable region.

[0647] 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, USDapartment of Health and Human Services, NIH Publication No. 91-3242); or (ii) the Chothia numbering scheme (hereinafter referred to as "Chothia CDRs") (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). 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 Duebel, 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. al., 1996, J. Mol. Biol., 262:732-745 (referred to herein as "AbM CDRs").See also, for example, Martin, A., "Protein Sequence and Structure Analysis of Antibody Variable Domains," in Antibody Engineering, Kontermann and Duebel, eds., Chapter 31, pp. 422-439, Springer-Verlag, Berlin (2001); or (v) according to the Contact numbering system, referred to herein as "Contact CDRs" (Contact definitions are based on analysis of available complex crystal structures (bioinf.org.uk / abs) (see, e.g., MacCallum et al., 1996, J. Mol. Biol., 262:732-745)).

[0648] As used herein, the terms "full-length antibody," "complete antibody," and "whole antibody" are used interchangeably to refer to an antibody in substantially its entire form, and not an antibody fragment as defined below. These terms specifically refer to antibodies having heavy chains that contain an Fc region.

[0649] An "antibody fragment" includes only a portion of an intact antibody, which portion retains at least one, two, three, and as many or all of the functions normally associated with that portion when present in an intact antibody. In one embodiment, an antibody fragment contains the antigen-binding site of an intact antibody and thus retains the ability to bind to an antigen. In another embodiment, an antibody fragment, e.g., 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, modulation of antibody half-life, 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 contain an 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.

[0650] "Polynucleotide" or "nucleic acid," used interchangeably herein, refers 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. 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 contain modified nucleotides, such as methylated nucleotides and their analogs. The nucleic acid molecule can be an aptamer.

[0651] The term "purification" refers to the separation of a substance (e.g., a compound, polynucleotide, protein, polypeptide, polypeptide composition) such that the substance of interest constitutes a majority of the sample. Typically, a substantially purified component of a sample comprises at least 50%, 80%-85%, 90-99%, such as at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, etc. 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.

[0652] The terms "treatment," "treating," and the like refer to achieving a desired pharmacological and / or physiological effect, such as a reduction in tumor burden. This effect can be preventative, meaning that a disease or its symptoms are completely or partially prevented, and / or therapeutic, meaning that a disease and / or harmful effects resulting from the disease are partially or completely cured. As used herein, "treatment" encompasses any treatment of a mammalian, particularly a human, disease, including (a) preventing the onset of a disease or disease symptoms in a subject who may be predisposed to the disease but has not yet been diagnosed with the disease (e.g., diseases that may be associated with or caused by a primary disease, such as liver fibrosis, which may occur in the setting 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., a reduction in tumor burden).

[0653] As used herein, the terms "individual," "host," "subject," and "patient" are used interchangeably and refer to animals, including, but not limited to, humans and non-human primates (including monkeys and humans), rodents (including rats and mice), cattle, horses, sheep, cats, dogs, etc. "Mammal" means a member or members of any mammalian species, including, by way of example, canines, felines, equines, bovines, ovines, rodents, etc., as well as primates (such as non-human primates) and humans. Non-human animal models, such as mammals, e.g., non-human primates, murines, lagomorphs, etc., can be used in experimental investigations.

[0654] A "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, condition, or disorder, is sufficient to effect treatment for the disease, condition, or disorder. The "therapeutically effective amount" will vary depending on the compound, the disease and its severity, and the age, weight, etc., of the patient being treated.

[0655] Unless otherwise specified, when a compound may 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, when a compound is described as a particular optical isomer, D- or L-, it is intended herein to include both optical isomers. For example, when a compound is described as having one of two tautomeric forms, it is intended herein to include both tautomers. 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 described herein may undergo epimerization in vivo. Thus, those skilled in the art will recognize that, for compounds that undergo epimerization in vivo, administering the compound in the (R) form is equivalent to administering the compound in the (S) form.

[0656] The present disclosure also encompasses all suitable isotopic variants of the compounds of the present disclosure, whether they are 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 that has the same atomic number but has an atomic mass that is different from the atomic mass that is usually or mainly found in nature.Examples of isotopes that can be incorporated into compounds of the present disclosure include isotopes of 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 and 131I. Certain isotopic variants of the compounds according to 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. For this purpose, 3 H, 14 C, and / or 18 Compounds labeled with the F isotope are suitable. Furthermore, the incorporation of an isotope (e.g., deuterium) may increase the metabolic stability of the compound, resulting in certain therapeutic benefits, such as an increased half-life in the body or a reduced effective dose. In some embodiments, hydrogen atoms in the compounds described herein may 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 amounts significantly greater than its natural abundance, unless otherwise specified. Isotopic variants of compounds according to the present disclosure can be prepared by a variety of methods, including, for example, those described below and in the Examples, using specific reagents and / or corresponding isotopic modifications of the starting compound.

[0657] Therefore, any embodiment described herein is meant to include salts, single stereoisomers, mixtures of stereoisomers, and / or isotopic forms of the compounds.

[0658] "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 preparing pharmaceutical compositions, and include excipients, diluents, carriers, and adjuvants that are acceptable for veterinary and human pharmaceutical use. As used herein and in the claims, "pharmaceutically acceptable excipients, diluents, carriers, and adjuvants" includes both one and more of such excipients, diluents, carriers, and adjuvants.

[0659] A "pharmaceutical composition" is meant to encompass compositions suitable for administration to a subject, such as a mammalian species, particularly a human. Generally, a "pharmaceutical composition" is sterile and preferably free of contaminants that could cause an undesirable reaction in the subject (e.g., the compound(s) in the pharmaceutical composition are of pharmaceutical grade). Pharmaceutical compositions can be designed to be administered to a subject or patient in need of treatment via a variety of routes of administration, including oral, buccal, rectal, parenteral, intraperitoneal, intradermal, intratracheal, intramuscular, and subcutaneous.

[0660] The term "pharmaceutically acceptable" means approved by a regulatory agency of the Federal or state government, or listed in the United States Pharmacopoeia, the European Pharmacopoeia, or other generally recognized pharmacopoeias, for use in animals, and especially in humans.

[0661] The term "pharmaceutically acceptable salt" refers to a salt suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic reaction, 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). Salts can be prepared in situ during the final isolation and purification of the conjugate compound, or can be prepared separately by reacting the free base functional group or groups of the compound with a suitable 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.

[0662] "Acyl" means 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)-, substituted hetero "C(O)-," "heterocyclyl-C(O)-," and "substituted heterocyclyl-C(O)-" groups, 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 CH3C(O)-.

[0663] The term "alkyl" refers to a branched or unbranched saturated hydrocarbon group (i.e., a monoradical) typically, but not necessarily, containing 1 to about 24 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, octyl, decyl, and the like, and cycloalkyl groups such as cyclopentyl and cyclohexyl. Generally, although not necessarily, alkyl groups herein can contain 1 to about 18 carbon atoms, and such groups can contain 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 group substituted with one or more substituents, including when two hydrogen atoms from the same carbon atom within the alkyl substituent are replaced, such as a carbonyl group (i.e., a substituted alkyl group can contain a -C(=O)- moiety). The terms "heteroatom-containing alkyl" and "heteroalkyl" refer to an alkyl substituent in which at least one carbon atom has been replaced with a heteroatom, as described in more detail below. Unless otherwise specified, the terms "alkyl" and "lower alkyl" include linear, branched, cyclic, unsubstituted, substituted, and / or heteroatom-containing alkyl or lower alkyl, respectively.

[0664] The term "substituted alkyl" is meant to include alkyl groups, 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- (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, thioheteroaryl. 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.

[0665] The term "alkenyl" refers to a linear, branched, or cyclic hydrocarbon group of 2 to about 24 carbon atoms containing at least one double bond, such as ethenyl, n-propenyl, isopropenyl, n-butenyl, isobutenyl, octenyl, decenyl, tetradecenyl, hexadecenyl, eicosenyl, tetracosenyl, etc. Generally, although not necessarily, an alkenyl group herein can 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 has been replaced with a heteroatom. Unless otherwise specified, the terms "alkenyl" and "lower alkenyl" include linear, branched, cyclic, unsubstituted, substituted, and / or heteroatom-containing alkenyl and lower alkenyl, respectively.

[0666] The term "alkynyl" refers to a linear or branched hydrocarbon group of 2 to 24 carbon atoms containing at least one triple bond, such as ethynyl, n-propynyl, etc. Generally, although not necessarily, alkynyl groups herein can contain from 2 to about 18 carbon atoms, and such groups may further contain from 2 to 12 carbon atoms. The term "lower alkynyl" refers to 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 in which at least one carbon atom has been replaced with a heteroatom. Unless otherwise specified, the terms "alkynyl" and "lower alkynyl" include linear, branched, unsubstituted, substituted, and / or heteroatom-containing alkynyl and lower alkynyl, respectively.

[0667] The term "alkoxy" refers to an alkyl group attached through a single terminal ether linkage, i.e., an "alkoxy" group is represented as -O-alkyl, where alkyl is as defined above. A "lower alkoxy" group refers to an alkoxy group containing from 1 to 6 carbon atoms and includes, for example, methoxy, ethoxy, n-propoxy, isopropoxy, t-butyloxy, and the like. As used herein, a substituent identified as a "C1-C6 alkoxy" or a "lower alkoxy" can contain, for example, 1 to 3 carbon atoms; as a further example, such a substituent can contain 1 or 2 carbon atoms (i.e., methoxy and ethoxy).

[0668] The term "substituted alkoxy" refers to substituted alkyl-O-, substituted alkenyl-O-, substituted cycloalkyl-O-, substituted cycloalkenyl-O-, and substituted alkynyl-O- groups, where substituted alkyl, substituted alkenyl, substituted cycloalkyl, substituted cycloalkenyl, and substituted alkynyl are as defined herein.

[0669] The term "aryl," unless otherwise specified, generally refers to an aromatic substituent containing 5 to 30 carbon atoms and including a single aromatic ring or multiple aromatic rings fused, directly linked, or indirectly linked to one another (different aromatic rings are bonded to a common group, such as a methylene or ethylene moiety). Aryl groups may contain, for example, 5 to 20 carbon atoms; as a further example, aryl groups may contain 5 to 12 carbon atoms. For example, aryl groups 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 aryl substituents in which at least one carbon atom has been replaced with a heteroatom, as described in more detail below. Aryl is a stable cyclic, heterocyclic, polycyclic, and polyheterocyclic unsaturated C-C ring aryl, exemplified by, but not limited to, phenyl, biphenyl, naphthyl, pyridyl, furyl, thiophenyl, imidazoyl, pyrimidinyl, and oxazoyl. 14 moieties, 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 specified, the term "aryl" includes unsubstituted, substituted, and / or heteroatom-containing aromatic substituents.

[0670] 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.

[0671] The term "alkylene" refers to polyvalent (e.g., dialkyl, trialkyl, tetraalkyl, etc.) groups. Unless otherwise specified, 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.

[0672] Similarly, the terms "alkenylene," "alkynylene," "arylene," "aralkylene," and "alkarylene" refer to the diradical alkenyl, alkynyl, aryl, aralkyl, and alkaryl groups, respectively.

[0673] In some embodiments, for example, in branched structures, "alkylene" refers to polyvalent (e.g., divalent alkyl groups, trivalent alkyl groups, tetravalent alkyl groups, etc.) Similarly, the terms "alkenylene," "alkynylene," "arylene," "aralkylene," and "alkarylene" can refer to polyvalent alkenyl, polyvalent alkynyl, polyvalent aryl, polyvalent aralkyl, and polyvalent alkaryl groups, respectively.

[0674] 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.

[0675] The terms "halo" and "halogen" are used in the conventional sense to refer to a chloro, bromo, fluoro, or iodo substituent.

[0676] "Carboxyl", "carboxy" or "carboxylate" refers to -CO2H or salts thereof.

[0677] "Cycloalkyl" refers to cyclic alkyl groups of 3 to 10 carbon atoms having single or multiple cyclic rings, including fused, bridged, and spirocyclic 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, and cyclooctyl, or multiple ring structures such as adamantanyl.

[0678] 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, carboxyalkyl, 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.

[0679] 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, linkage, 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 a heteroatom-containing alkyl substituent, the term "heterocycloalkyl" refers to a heteroatom-containing cycloalkyl substituent, the terms "heterocyclic" or "heterocycle" refer to a heteroatom-containing cyclic substituent, the terms "heteroaryl" and "heteroaromatic" refer to heteroatom-containing "aryl" and "aromatic" substituents, respectively, and so forth. Examples of heteroalkyl groups include alkoxyaryl, alkylsulfanyl-substituted alkyl, N-alkylated aminoalkyl, 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.

[0680] "Heteroaryl" refers to an aromatic group having 1 to 15 carbon atoms, e.g., 1 to 10 carbon atoms, and 1 to 10 heteroatoms selected from the group consisting of oxygen, nitrogen, and sulfur within the ring. Such heteroaryl groups can have a single ring (e.g., pyridinyl, imidazolyl, or furyl) or multiple condensed rings within the ring system (e.g., groups such as indolizinyl, quinolinyl, benzofuran, benzimidazolyl, or benzothienyl), where at least one ring within the ring system is aromatic and the point of attachment is through an aromatic ring atom. 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. This term includes, for example, pyridinyl, pyrrolyl, indolyl, thiophenyl, and furanyl. Unless otherwise limited by the definition of a heteroaryl substituent, such heteroaryl groups can be optionally substituted with one to five substituents, or one to three 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.

[0681] 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, and in fused ring systems, one or more of the rings may be cycloalkyl, heterocycloalkyl, aryl, or heteroaryl, so long as the point of attachment is through a non-aromatic ring. In certain embodiments, the nitrogen and / or sulfur atoms of a heterocyclic group are optionally oxidized to form an N-oxide, -S(O)-, or -SO2- moiety.

[0682] Examples of heterocycles and heteroaryls are 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 ... including, but 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.

[0683] Unless otherwise limited by the definition of a heterocycle substituent, such a heterocycle group can be optionally substituted with one to five, or one to three, 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.

[0684] "Hydrocarbyl" refers to a monovalent hydrocarbyl radical containing 1 to about 30 carbon atoms, preferably 1 to about 24 carbon atoms, further 1 to about 18 carbon atoms, further about 1 to 12 carbon atoms, and includes linear, branched, cyclic, saturated, and unsaturated species such as alkyl, alkenyl, and aryl groups. Hydrocarbyls can 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 specified, the term "hydrocarbyl" is intended to include substituted hydrocarbyl moieties and / or heteroatom-containing hydrocarbyl moieties.

[0685] "Substituted," as in "substituted hydrocarbyl," "substituted alkyl," "substituted aryl," and the like, referred to in some of the above definitions, 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 alkenyl, 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 above hydrocarbyl moieties may be further substituted with one or more functional groups or additional hydrocarbyl moieties, such as those specifically enumerated. Unless otherwise specified, all groups described herein are understood to include substituted and / or heteroatom-containing moieties in addition to unsubstituted groups.

[0686] "Sulfonyl" refers to 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 groups, where alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, and substituted heterocyclic are as defined herein. Sulfonyl includes, for example, methyl-SO-, phenyl-SO-, and 4-methylphenyl-SO-.

[0687] "Functional group" refers to a group selected from the group consisting of halo, hydroxyl, sulfhydryl, C1-C24 alkoxy, C2-C24 alkenyloxy, C2-C24 alkynyloxy, C5-C20 aryloxy, acyl (including 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( 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), carbamide (-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 aryl It refers to chemical groups such as amide (-NH-(CO)-aryl), imino (-CR=NH, R=hydrogen, C1-C24 alkyl, C5-C20 aryl, C6-C20 alkaryl, C6-C20 aralkyl, etc.), alkylimino (-CR=N(alkyl), 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-C 24alkylsulfanyl (-S-alkyl, also referred to as alkylthio), arylsulfanyl (-S-aryl, also referred to as arylthio), 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. Additionally, the foregoing functional groups may be further substituted, where the particular group permits, with one or more additional functional groups, or one or more hydrocarbyl moieties, such as those specifically enumerated above.

[0688] "Linkage" or "linker," as used herein, refers to a "linking group," "linker moiety," or the like, that connects two groups via a covalent bond. Linkers can be linear, branched, cyclic, or single atom. Examples of such linking groups include alkyl, alkenylene, alkynylene, arylene, alkarylene, aralkylene, and linking moieties containing 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 circumstances, 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 are no more than one, two, or three unsaturated bonds in the linker backbone. The linker may include one or more substituents, such as, for example, alkyl, aryl, or alkenyl groups. The linker may be a poly(ethylene glycol) unit (e.g., —(CH—CH—O)—), an ether, a thioether, an amine, an alkyl (e.g., (C1-C 12 ) alkyl), which may be linear or branched, such as, but not limited to, 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, for example, an aryl, heterocycle, or cycloalkyl group, where two or more atoms of the cyclic group, for example, 2, 3, or 4 atoms, are included in the backbone. The linker may be cleavable or non-cleavable. Any convenient orientation and / or connection between the linker and the linked group may be used.

[0689] When the term "substituted" appears before a list of possible substituents, it is intended that the term apply to all members of that group. For example, the phrase "substituted alkyl and aryl" is interpreted as "substituted alkyl and substituted aryl."

[0690] Further to the disclosure herein, the term "substituted," when used to modify a particular group or radical, can also mean that one or more hydrogen atoms of the particular group or radical are each, independently of one another, replaced with the same or different substituents, as defined below.

[0691] In addition to the groups disclosed herein for individual terms, one or more hydrogens on a saturated carbon atom in a particular group or radical (any two hydrogens on a single carbon can be ═O, ═NR 70 , =N-OR 70 , =N2 or =S) is substituted with -R unless otherwise specified. 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; and each R 70 are independently hydrogen or R 60 and each R 80 is independently R 70 or, alternatively, 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 of the same or different additional heteroatoms selected from the group consisting of O, N, and S,...

Claims

1. A conjugate comprising a ligand moiety X conjugated to a target binding moiety Y via a linker L, the ligand moiety X binds to a lysosomal targeting molecule extracellularly; said target binding moiety Y binds to a target molecule extracellularly; the target binding moiety Y dissociates from the target molecule in the endosome; The conjugate is externalized from the cell.

2. 2. The conjugate of claim 1, wherein the ligand moiety X remains bound to the lysosomal targeting molecule within the endosome.

3. 3. The conjugate of claim 1 or 2, wherein the target-binding moiety Y binds to FcRn in an endosome.

4. The conjugate of any one of claims 1 to 3, wherein the conjugate dissociates from the lysosome-targeting molecule within an endosome.

5. The conjugate of claim 1 , wherein the lysosomal targeting molecule is ASGPR.

6. 2. The conjugate of claim 1, wherein Y is an antibody or an antibody fragment.

7. The conjugate of claim 2 , wherein the conjugate is externalized from the cell via the lysosomal targeting molecule.

8. 3. The conjugate of claim 2, wherein the ligand moiety X has equal binding affinity for the lysosomal targeting molecule extracellularly and in endosomes.

9. 3. The conjugate of claim 2, wherein the ligand moiety X has equal binding affinity for the lysosome-targeting molecule at extracellular and intraendosomal pH.

10. The ligand moiety X is a ligand of extracellular Ca ２＋ Concentration and intraendosomal Ca ２＋ The conjugate of claim 2, having equal binding affinity for the lysosomal targeting molecule at different concentrations.

11. The conjugate of claim 3 , wherein the conjugate is externalized from the cell via FcRn.

12. 5. The conjugate of claim 4, wherein the target-binding moiety Y binds to FcRn in an endosome.

13. The conjugate of claim 12, wherein the conjugate is externalized from the cell via FcRn.

14. 13. The conjugate of claim 12, wherein the ligand moiety X has a higher binding affinity for the lysosome-targeted molecule outside the cell than within the endosome.

15. 13. The conjugate of claim 12, wherein the ligand moiety X has a higher binding affinity for the lysosome-targeted molecule at extracellular pH than at intraendosomal pH.

16. The ligand moiety X is a ligand of intraendosomal Ca ２＋ Extracellular Ca concentration ２＋ The conjugate of claim 12, having a high binding affinity for the lysosome-targeting molecule at a concentration of

17. 13. The conjugate of claim 12, wherein the target-binding moiety Y has a higher binding affinity for FcRn in endosomes than outside the cell.

18. 13. The conjugate of claim 12, wherein the target-binding moiety Y has a higher binding affinity for FcRn at endosomal pH than at extracellular pH.

19. 13. The conjugate of claim 12, wherein the target binding moiety Y has enhanced binding to FcRn compared to wild type at endosomal pH.

20. 13. The conjugate of claim 12, wherein the target-binding moiety Y has approximately the same binding affinity for FcRn extracellularly as that of wild-type IgG for FcRn extracellularly.

21. 13. The conjugate of claim 12, wherein the target-binding moiety Y has approximately the same binding affinity to FcRn as wild-type IgG at pH 7.

4.

22. 13. The conjugate of claim 12, wherein Y is an antibody.

23. 2. The conjugate of claim 1, wherein the target-binding moiety Y has a higher binding affinity for the target molecule outside the cell than inside the endosome.

24. 2. The conjugate of claim 1, wherein the target-binding moiety Y has a higher binding affinity for the target molecule at extracellular pH than at endosomal pH.

25. The target binding moiety Y has a K of 2:1 to 10,000:1 with respect to the target molecule at pH 6.0:7.4 Ｄ The conjugate of claim 1 having a ratio:

26. The target binding moiety Y has a higher k ｏｆｆ The conjugate of claim 1 having a ratio of

27. 10. The conjugate of claim 1, wherein the conjugate can be cycled over a period of several hours to several days.

28. 2. The conjugate of claim 1, wherein the target binding moiety Y is an antibody or antibody fragment mutated from the wild type by one or more histidine substitutions.

29. The conjugate has the formula (I'): 【Chemical 325】 or a prodrug thereof, or a pharmaceutically acceptable salt thereof; During the ceremony, n is 1 to 500; m is 1 to 20; X is the ligand moiety; 10. A conjugate according to any one of the preceding claims, wherein Y is the target binding moiety.

30. 10. The conjugate of any one of the preceding claims, wherein the lysosomal targeting molecule is a cell surface receptor that allows internalization of the conjugate.

31. 10. The conjugate of any one of the preceding claims, wherein the lysosomal targeting molecule is selected from the asialoglycoprotein receptor (ASGPR), the cation-independent mannose-6-phosphate receptor (CI-M6PR, also referred to herein as M6PR), the folate receptor, LDLR, CD63, sortilin, IFITM3, molecules in the endosomal / lysosomal pathway, LIMP-1, and LIMP-2.

32. 10. A conjugate according to any one of the preceding claims, wherein X is a moiety that binds to ASGPR or M6PR.

33. 10. A conjugate according to any one of the preceding claims, wherein X is a moiety that binds to ASGPR.

34. The conjugate has the formula (I'): 【Chemistry 326】 or a prodrug thereof, or a pharmaceutically acceptable salt thereof; During the ceremony, n is 1 to 500; m is 1 to 20; L is a linker, X is a group represented by formula (II) 【Chemistry 327】 an asialoglycoprotein receptor (ASGPR) binding portion of the formula: R １ Is, -Z １ -*, -H, -OH, optionally substituted (C １ ~C ６ ) alkyl, —OCH ３ , -OCH ２ CH═CH, optionally substituted —S—(C １ ~C ６ ) selected from alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted —S-aryl, and optionally substituted —S-heteroaryl; R ２ Is, -Z １ -*, -NHCOCH ３ , -NHCOCF ３ , -NHCOCH ２ CF ３ , —OH, —NHR, and optionally substituted triazole; R ６ Is, -Z １ -*, -OH, -OR, optionally substituted (C １ ~C ６ ) alkyl, —OC(O)R, —C(O)NHR, —NR ｘｘ R ｙｙ , optionally substituted aryl, optionally substituted heteroaryl, —NHCOR, and —NRCOR; Each R is independently an optionally substituted (C １ ~C ６ ) alkyl, optionally substituted aryl, or optionally substituted heteroaryl; R ｘｘ and R ｙｙ are independently H, optionally substituted (C １ ~C ６ ) alkyl, or R ｘｘ and R ｙｙ can cyclize to form an optionally substituted heterocyclyl; In the formula, R １ , R ２ , and R ６ One of them is -Z １ -*, and "*" is Z １ and the linker (L), R ３ and R ４ are each independently H, or a pro-moiety, or R ３ and R ４ are linked in a ring to form a pro-moiety, R １１ is H or a bridging moiety connecting the 5-carbon to the 1-carbon of the ring, Z １ Is, -Z １１ -, -Z １１ -A １ -, -A ２ -, -NR ２１ CO-, -CONR ２１ -, -NR ２１ SO ２ -, -SO ２ NR ２１ -, -NR ２１ C(=O)NR ２１ - and -NR ２１ C(=S)NR ２１ - is a linking moiety selected from -Z １１ - is -O-, -S-, -N(R ２１ ) - or -C(R ２２ ) ２ where R １ Ga-Z １ - *, Z １ Ga-Z １１ If -, then -Z １１ - is not -O-, -A １ - and - A ２ - is an optionally substituted heterocyclylene, an optionally substituted arylene, or an optionally substituted heteroarylene; Each R ２１ is H, optionally substituted (C １ ~C ６ ) independently selected from alkyl, —COR, and optionally substituted heteroaryl; Each R ２２ is H, halogen, and optionally substituted (C １ ~C ６ 34. The conjugate of claim 33, wherein each of the groups is independently selected from: alkyl;

35. -L-Y is 【Chemical 328】 Including, In the formula, R Ｙ teeth, 【Chemistry 329】 35. The conjugate of claim 34, wherein:

36. X is a group represented by formula (a-II) 【Chemistry 330】 36. The conjugate of claim 34 or 35, wherein:

37. R １ Is, -Z １ -*, -H, or (C １ ~C ６ 37. The conjugate of any one of claims 34 to 36, wherein:

38. R ２ Is, -Z １ -* or -NHCOCH ３ The conjugate according to any one of claims 34 to 36, wherein

39. R ３ and R ４ The conjugate of any one of claims 34 to 38, wherein each is -H.

40. 33. The conjugate of claim 32, wherein X is a moiety that binds to M6PR.

41. The conjugate has the formula (IV): 【Chemistry 331】 wherein: W is a non-hydrolyzable hydrophilic head group; Z １ is optionally substituted (C １ ~C ３ ) alkylene and optionally substituted ethenylene; Z ２ is S, NR ２１ and C(R ２２ ) ２ wherein each R ２１ is H and optionally substituted (C １ ~C ６ ) alkyl, and each R ２２ is H, halogen and optionally substituted (C １ ~C ６ ) alkyl; each A is independently an optionally substituted aryl or heteroaryl linking moiety; Each Z ３ is independently a linking moiety.

42. Z ２ The conjugate of claim 41, wherein is S.

43. 43. The conjugate of claim 41 or 42, wherein W is a phosphonate, thiophosphonate, carboxylic acid, or malonic acid, or a salt thereof.

44. X is as follows: 【Chemistry 332】 In the formula, R ａ , R ｂ , R ｃ and R ｄ The conjugate of any one of claims 45 to 47, wherein: is independently H or F.

45. X is as follows: 【Chemical 333】 In the formula, R ａ , R ｂ , R ｃ and R ｄ The conjugate of any one of claims 41 to 44, wherein

46. 46. ​​The conjugate of any one of claims 41 to 45, wherein A is optionally substituted aryl or optionally substituted heteroaryl, preferably A is independently selected from optionally substituted phenyl, optionally substituted pyridyl, optionally substituted biphenyl, optionally substituted naphthalene, optionally substituted triazole and optionally substituted phenylene-triazole.

47. 10. The conjugate of any one of the preceding claims, wherein L comprises 10 to 60 consecutive branched or straight chain atoms.

48. L is a group represented by the formula (IIb'): 【Chemistry 334】 wherein: n is 1, 2, or 3; L １ ~L ６ are each independently a linking moiety, and together they form Z １ and forming a linear or branched linker between Y; a, b, c, d, and e are each independently 1, 2, 3, 4, or 5; ** is Z １ X's L via １ represents the attachment point to 10. A conjugate according to any one of the preceding claims, wherein *** represents the point of attachment to Y.

49. Each L １ ~L ５ is -C １～２０ -Alkylene-, -NHC(O)-C １～６ -Alkylene-, -C(O)NH-C １～６ -Alkylene-, -NH-C １～６ -Alkylene-, -NHC(O)NH-C １～６ -Alkylene-, -NHC(S)NH-C １～６ -Alkylene-, -C １～６ -Alkylene-NHC(O)-, -C １～６ -Alkylene-C(O)NH-, -C １～６ -Alkylene-NH-, -C １～６ -Alkylene-NHC(O)NH-, -C １～６ -Alkylene-NHC(S)NH-, -O(CH ２ ) ｐ -, -(OCH ２ CH ２ ) ｐ -, -NHC(O)-, -C(O)NH-, -NHS(O) ２ -, -S(O) ２ NH-, -C(O)-, -S(O) ２ independently comprise one or more linking moieties independently selected from -, -O-, -S-, monocyclic heteroaryl, monocyclic aryl, monocyclic heterocycle, monocyclic carbocycle, an amino acid residue, -NH-, and -NMe-, wherein L １ ~L ５ are each independently optionally substituted with 1 to 5 halo; each p is independently 1 to 50; L ６ is -C １～２０ -Alkylene-, -NR １６ C(O)-C １～６ -Alkylene-, -C(O)NR １６ -C １～６ -Alkylene-, -NR １６ -C １～６ -Alkylene-, -NR １６ C(O)NR １６ -C １～６ -Alkylene-, -NR １６ C(S)NR １６ -C １～６ -Alkylene-, -C １～６ -Alkylene-NR １６ C(O)-, -C １～６ -Alkylene-C(O)NR １６ -, -C １～６ -Alkylene-NR １６ -, -C １～６ -Alkylene-NR １６ C(O)NR １６ -, -C １～６ -Alkylene-NR １６ C(S)NR １６ -, -O(CH ２ ) ｐ -, -(OCH ２ CH ２ ) ｐ -, -NR １６ C(O)-, -C(O)NR １６ -, -NHS(O) ２ -, -S(O) ２ NH-, -C(O)-, -S(O) ２ a linking group comprising one or more linking moieties independently selected from -, -O-, -S-, monocyclic heteroaryl, monocyclic aryl, monocyclic heterocycle, an amino acid residue, or -NR16-; Each R １６ are independently —H, optionally substituted (C １ ~C ６ 49. The conjugate of claim 48, wherein: R is an alkyl, an optionally substituted aryl, an optionally substituted monocyclic heteroaryl, or a monocyclic heteroaryl.

50. Each L １ ~L ５ is -C １～２０ -Alkylene-, -NHC(O)-C １～６ -Alkylene-, -C(O)NH-C １～６ -Alkylene-, -NH-C １～６ -Alkylene-, -NHC(O)NH-C １～６ -Alkylene-, -NHC(S)NH-C １～６ -Alkylene-, -C １～６ -Alkylene-NHC(O)-, -C １～６ -Alkylene-C(O)NH-, -C １～６ -Alkylene-NH-, -C １～６ -Alkylene-NHC(O)NH-, -C １～６ -Alkylene-NHC(S)NH-, -O(CH ２ ) ｐ -, -(OCH ２ CH ２ ) ｐ -, -NHC(O)-, -C(O)NH-, -NHS(O) ２ -, -S(O) ２ NH-, -C(O)-, -S(O) ２ independently selected from -, -O-, -S-, monocyclic heteroaryl, monocyclic aryl, monocyclic heterocycle, monocyclic carbocycle, an amino acid residue, -NH-, and -NMe-; １ ~L ５ are each independently optionally substituted with 1 to 5 halo; each p is independently 1 to 50; L ６ teeth, 【Chemistry 335】 and In the formula, R ｚ teeth, 【Chemistry 336】 49. The conjugate of claim 48, wherein:

50. 10. A method for degrading a target molecule in a subject in need thereof, the method comprising administering to the subject an effective amount of a conjugate according to any one of the preceding claims.

51. 51. The method of claim 50, wherein at least 90% of the target is degraded 4 days after administration.

52. 51. The method of claim 50, wherein at least 90% of the target is degraded 7 days after administration.

53. 51. The method of claim 50, wherein the extracellular concentration of the target is substantially maintained at an amount at least 90% lower than the initial extracellular concentration of the target prior to administering an effective amount of the conjugate, for a period of at least 4 days.

54. 51. The method of claim 50, wherein the period is seven days or longer.

55. 55. The method of any one of claims 50 to 54, wherein the superstoichiometric target:conjugate ratio is reduced.

56. 56. The method of claim 55, wherein the ratio is at least about 5.

57. 57. The method of any one of claims 50 to 56, wherein the target binding moiety Y has a higher binding affinity for the target molecule outside the cell than inside the endosome.

58. 58. The method of any one of claims 50 to 57, wherein the target is IgE.

59. 1. A method for degrading a target molecule in a subject in need thereof, comprising: a means for binding to a lysosomal targeting molecule extracellularly; a means for binding to a target molecule extracellularly; and means for dissociating from said target molecule in an endosome, The method wherein the conjugate is externalized from the cell.

60. 60. The method of claim 59, wherein said means for binding to a lysosomal targeting molecule remains bound to said lysosomal targeting molecule within an endosome.

61. 61. The method of claim 59 or 60, wherein the means for binding to a target molecule also binds to FcRn in an endosome.

62. 62. The method of claim 61, wherein the conjugate dissociates from the lysosome-targeting molecule within an endosome.

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