Cell surface receptor binding compounds and conjugates

Abstract

To provide cell surface receptor-binding compounds and conjugates. [Solution] This disclosure provides a class of compounds comprising a ligand moiety that specifically binds to cell surface receptors such as mannose-6-phosphate receptor (M6PR) or cell surface asialoglycoprotein receptor (ASGPR). Cell surface M6PR or ASGPR-binding compounds can trigger the receptor to take up the bound compound into the cell. The ligand moieties of this disclosure can be conjugated to various parts of interest without affecting the specific binding to cell surface receptors, e.g., M6PR or ASGPR, and their function. Compounds that are conjugates of the ligand moiety conjugated to biomolecules such as antibodies are also provided, and the conjugate can use cellular pathways to remove specific proteins of interest from the cell surface or extracellular environment.

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims the interests of U.S. applications 62 / 959,877, 62 / 959,862, 62 / 959,882, 63 / 043,749, 63 / 043,752, and 63 / 043,754, filed on 24 June 2020, which are incorporated herein by reference in their entirety.

[0002] Reference to electronically submitted sequence list This application was prepared on 17 June 2020 and is incorporated by reference in a text file titled 47970WO Seqlist, which was submitted with this application and has a size of 15,951 bytes. [Background technology]

[0003] introduction Many therapeutic agents act by binding to functionally important sites on target proteins and thereby regulating their activity, or by recruiting immune effectors, as is the case with many monoclonal antibody drugs. However, there is an undeveloped reservoir of these proteins that are considered "undruggable" because they do not readily conform to currently available therapeutic targeting approaches. Therefore, there is a need for therapies that can target a wider range of proteins.

[0004] Mannose-6-phosphate is a monosaccharide ligand that plays a crucial role in the intracellular retention and secretion of lysosomal hydrolases to which it binds. When this sugar residue is incorporated into newly synthesized enzymes, the enzymes are transported from the Golgi apparatus to lysosomes, where they are activated. Cell surface mannose-6-phosphate receptors (M6PRs), bound to the cell membrane, play a role in many biological processes, including the secretion and internalization of such lysosomal enzymes. Endocytosis by M6PRs enables the internalization of compounds containing mannose-6-phosphate (M6P) ligands into cells and their transport to lysosomes.

[0005] Alternative ligands that provide binding to cell surface M6PR and subsequent transport across the cell membrane are of great interest. [Overview of the project] [Means for solving the problem]

[0006] This disclosure provides a class of compounds comprising ligand moieties that specifically bind to cell surface receptors. In some embodiments, the ligand moiety binds to mannose-6-phosphate receptors (M6PR). In some embodiments, the ligand moiety binds to cell surface asialoglycoprotein receptors (ASGPR). Cell surface M6PR or ASGPR-binding compounds can trigger the receptor to take up the bound compound into the cell. The ligand moieties of this disclosure can be conjugated to various parts of interest without affecting the specific binding to cell surface receptors, e.g., M6PR or ASGPR, and their function. Compounds that are conjugates of ligand moieties conjugated to biomolecules such as antibodies are also provided, and the conjugates can utilize cellular pathways to remove specific proteins of interest from the cell surface or extracellular environment. For example, the conjugates described herein can sequester and / or degrade target molecules of interest within cellular lysosomes. Compositions comprising such conjugates, methods for targeting polypeptides of interest for sequester and / or lysosomal degradation using the conjugates, and methods for treating disorders or diseases using the conjugates are also provided herein.

[0007] A first aspect of this disclosure relates to a cell surface mannose-6-phosphate receptor (M6PR) binding compound of formula (XI): [ka] or a salt thereof, in the formula, Each W is independently a hydrophilic head group. each Z 1 However, independently, they are selected from arbitrarily substituted (C1-C3) alkylenes and arbitrarily substituted ethenylenes. each Z 2 However, independently, O, S, NR 21 , and C(R 22 ) Selected from 2, each R 21 However, each R is independently selected from H and optionally substituted (C1-C6) alkyl groups. 22However, independently selected from H, halogens (e.g., F), and optionally substituted (C1-C6) alkyl groups, Each Ar is independently an optionally substituted aryl or heteroaryl linking moiety (e.g., an optionally substituted monocyclic or bicyclic aryl or heteroaryl), each Z 3 However, it is an independent connecting part, n is between 1 and 500. L is the linker, Y is the part of interest.

[0008] A second aspect of this disclosure relates to a cell surface receptor-binding conjugate of formula (I): [ka] or a salt thereof During the ceremony, X is a portion that binds to the asialoclycoprotein receptor (ASGPR) on the cell surface, or to the mannose-6-phosphate receptor (M6PR) on the cell surface. n is between 1 and 500 (for example, n is between 1 and 20, 1 and 10, 1 and 6, or 1 and 5), L is the linker, Y is a biomolecule that specifically binds to target proteins.

[0009] In some embodiments of formula (I), Y is an antibody or antibody fragment that specifically binds to a target protein, and the compound is the compound of formula (V): [ka] or a pharmaceutically acceptable salt thereof During the ceremony, n is between 1 and 20. m is the average load from 1 to 80. Ab is an antibody or antibody fragment that specifically binds to a target protein. Z is the residue portion resulting from the covalent bonding of a chemoselective ligation group to the compatible group of Ab.

[0010] A third aspect of this disclosure is a method for internalizing a target protein into a cell containing a cell surface receptor selected from M6PR and ASGPR, comprising contacting a cell sample containing the cell and the target protein with an effective amount of a compound or conjugate (e.g., as described herein), which specifically binds to the target protein and specifically binds to the cell surface receptor to promote cellular uptake of the target protein.

[0011] A fourth aspect of this disclosure is a method for reducing the level of a target protein in a biological system, comprising contacting the biological system with an effective amount of a compound or conjugate (e.g., as described herein) that specifically binds to the target protein and specifically binds to cell surface receptors of cells in the biological system, thereby promoting the cellular uptake and degradation of the target protein. [Brief explanation of the drawing]

[0012] [Figure 1] A typical SEC chromatogram of matsuzumab-(compound A) conjugate. [Figure 2] Native mass spectrometry (MS) analysis of deglycosylated matsuzumab and matsuzumab-(compound A) conjugate. [Figure 3] A typical SEC chromatogram of matsuzumab-(compound I-7) conjugate. [Figure 4] Native MS analysis of deglycosylated matsuzumab and matsuzumab-(compound I-7) conjugate. [Figure 5] A typical SEC chromatogram of the atezolizumab-(compound A) conjugate. [Figure 6] Native MS analysis of deglycosylated atezolizumab and atezolizumab-(compound A) conjugate. [Figure 7] A typical SEC chromatogram of the cetuximab-(compound A) conjugate. [Figure 8] Native MS analysis of deglycosylated cetuximab and cetuximab-(compound A) conjugate. [Figure 9] A typical SEC chromatogram of the cetuximab-(compound I-7) conjugate. [Figure 10] Native MS analysis of deglycosylated cetuximab and cetuximab-(compound I-7) conjugate. [Figure 11] A representative SEC chromatogram of the anti-PD-L1 antibody (29E.2A3)-(compound A) conjugate. [Figure 12-1] Native MS analysis of deglycosylated anti-PD-L1 antibody (29E.2A3) and anti-PD-L1 antibody (29E.2A3)-(compound A) conjugate. [Figure 12-2] Native MS analysis of deglycosylated anti-PD-L1 antibody (29E.2A3) and anti-PD-L1 antibody (29E.2A3)-(compound A) conjugate. [Figure 13] A typical SEC chromatogram of the IgG2a-UNLB-(compound I-7) conjugate. [Figure 14] Native MS analysis of deglycosylated IgG2a-UNLB and IgG2a-UNLB-(compound I-7) conjugate. [Figure 15] Time-course activity of cetuximab-(compound A) and cetuximab-(compounds I-7) conjugates in surface EGFR levels of Hela parental cells and M6PR KO cells, as measured by surface staining. [Figure 16] Time-dependent activity of matsuzumab-(compound A) and matsuzumab-(compounds I-7) conjugates in surface EGFR levels of Hela parental and M6PR KO cells, as measured by surface staining. [Figure 17]Dose-response of cetuximab-(compound A), cetuximab-(compounds I-7), matsuzumab-(compound A), and matsuzumab-(compounds I-7) conjugates in total EGFR levels of Hela parental and M6PR KO cells as measured by intracellular Western blotting. [Figure 18] Time-dependent activity of cetuximab-(compound A), cetuximab-(compounds I-7), matsuzumab-(compound A), and matsuzumab-(compound I-7) conjugates in relative EGFR normalization levels of Hela parent and M6PR KO cells. [Figure 19A] Binding affinity of matsuzumab conjugated to M6PR with unlabeled control (Figure 19A), compound I-7 (Figure 19B), compound I-8 (Figure 19C), compound I-9 (Figure 19D), compound I-11 (Figure 19E), and compound I-12 (Figure 19F) to M6PR. Binding to M6PR was determined by ELISA. Compound I-7 (dar8) and compound I-11 (dar4) showed the highest and lowest binding affinity, respectively. RFU: Relative fluorescence units. [Figure 19B] Binding affinity of matsuzumab conjugated to M6PR with unlabeled control (Figure 19A), compound I-7 (Figure 19B), compound I-8 (Figure 19C), compound I-9 (Figure 19D), compound I-11 (Figure 19E), and compound I-12 (Figure 19F) to M6PR. Binding to M6PR was determined by ELISA. Compound I-7 (dar8) and compound I-11 (dar4) showed the highest and lowest binding affinity, respectively. RFU: Relative fluorescence units. [Figure 19C] Binding affinity of matsuzumab conjugated to M6PR with unlabeled control (Figure 19A), compound I-7 (Figure 19B), compound I-8 (Figure 19C), compound I-9 (Figure 19D), compound I-11 (Figure 19E), and compound I-12 (Figure 19F) to M6PR. Binding to M6PR was determined by ELISA. Compound I-7 (dar8) and compound I-11 (dar4) showed the highest and lowest binding affinity, respectively. RFU: Relative fluorescence units. [Figure 19D]Binding affinity of matsuzumab conjugated to M6PR with unlabeled control (Figure 19A), compound I-7 (Figure 19B), compound I-8 (Figure 19C), compound I-9 (Figure 19D), compound I-11 (Figure 19E), and compound I-12 (Figure 19F) to M6PR. Binding to M6PR was determined by ELISA. Compound I-7 (dar8) and compound I-11 (dar4) showed the highest and lowest binding affinity, respectively. RFU: Relative fluorescence units. [Figure 19E] Binding affinity of matsuzumab conjugated to M6PR with unlabeled control (Figure 19A), compound I-7 (Figure 19B), compound I-8 (Figure 19C), compound I-9 (Figure 19D), compound I-11 (Figure 19E), and compound I-12 (Figure 19F) to M6PR. Binding to M6PR was determined by ELISA. Compound I-7 (dar8) and compound I-11 (dar4) showed the highest and lowest binding affinity, respectively. RFU: Relative fluorescence units. [Figure 19F] Binding affinity of matsuzumab conjugated to M6PR with unlabeled control (Figure 19A), compound I-7 (Figure 19B), compound I-8 (Figure 19C), compound I-9 (Figure 19D), compound I-11 (Figure 19E), and compound I-12 (Figure 19F) to M6PR. Binding to M6PR was determined by ELISA. Compound I-7 (dar8) and compound I-11 (dar4) showed the highest and lowest binding affinity, respectively. RFU: Relative fluorescence units. [Figure 20A] Serum PK analysis of individual rIgG1 antibody conjugates. Intracellular levels of aIgG2a conjugated with compounds I-7 (dar8) and (dar4) (Figure 20A), aIgG2a conjugated with compound I-11 and aIgG2a conjugated with compound I-11 (Figure 20B), and aIgG2a conjugated with compound I-9 and aIgG2a conjugated with compound I-12 (Figure 20C) in mouse serum were measured using ELISA at 0.5, 1, 2, 6, and 24 hours. [Figure 20B]Serum PK analysis of individual rIgG1 antibody conjugates. Intracellular levels of aIgG2a conjugated with compounds I-7 (dar8) and (dar4) (Figure 20A), aIgG2a conjugated with compound I-11 and aIgG2a conjugated with compound I-11 (Figure 20B), and aIgG2a conjugated with compound I-9 and aIgG2a conjugated with compound I-12 (Figure 20C) in mouse serum were measured using ELISA at 0.5, 1, 2, 6, and 24 hours. [Figure 20C] Serum PK analysis of individual rIgG1 antibody conjugates. Intracellular levels of aIgG2a conjugated with compounds I-7 (dar8) and (dar4) (Figure 20A), aIgG2a conjugated with compound I-11 and aIgG2a conjugated with compound I-11 (Figure 20B), and aIgG2a conjugated with compound I-9 and aIgG2a conjugated with compound I-12 (Figure 20C) in mouse serum were measured using ELISA at 0.5, 1, 2, 6, and 24 hours. [Figure 21] Intracellular uptake of anti-IgG2a conjugates in Jurkat cells. Conjugates were detected using an antibody conjugated with Alex488, and intracellular fluorescence levels were determined by FACS at 1 hour and 24 hours. [Figure 22] Intracellular uptake of anti-IgG2a is measured as a percentage of the uptake of the aIgG2a conjugate compound I-7(dar8) after 24 hours, and conjugation to Jurkat cells at 10 nM is performed. [Figure 23] Graphs showing the results of M6PR binding assays for various antibody conjugates of exemplary compounds with varying DAR loadings. [Figure 24] The graph of cell fluorescence versus antibody conjugate concentrations shows that various antibody conjugates of exemplary M6PR-binding compounds exhibited robust uptake into Jurkat cells after 1 hour of incubation. [Figure 25]The graph of cell fluorescence versus antibody conjugate concentrations shows that various antibody conjugates of exemplary M6PR or ASGPR-conjugated compounds showed robust uptake into HepG2 cells after 1 hour of incubation. [Figure 26] A graph showing CI-M6PR-dependent cellular uptake of exemplary antibody conjugates in wild-type (WT) K562 cells versus CI-M6PR knockout (KO) cells. [Modes for carrying out the invention]

[0013] As summarized above, this disclosure provides a class of compounds comprising ligand moieties that specifically bind to cell surface receptors. Also provided herein are conjugates comprising a moiety X that binds to such cell surface receptors, e.g., internalized cell surface receptors, for sequesteration and / or lysosomal degradation. In certain embodiments, the cell surface receptor is a mannose-6-phosphate receptor (M6PR). In certain embodiments, the cell surface receptor is an asialoglycoprotein receptor (ASGPR).

[0014] This disclosure concerns compounds of formula (I): [ka] or a salt thereof, in the formula, X is a portion that binds to a cell surface receptor selected from M6PR and ASGPR (for example, as described herein), n is between 1 and 500. L is a linker of a defined length (e.g., monovalent or polyvalent, as described herein), Y is the part of interest (for example, as described herein).

[0015] The compounds, conjugates, and methods of this disclosure are described in more detail below. Specific classes of M6PR-binding compounds are described. Biomolecular conjugates comprising a cell surface receptor-binding moiety (X) that binds to M6PR or ASGPR are also described. Linkers (L) and moieties of interest (Y) that find applications in M6PR-binding compounds, as well as biomolecular conjugates, are also described. Methods by which the compounds and conjugates of this disclosure find applications are also described.

[0016] M6PR binding compound As summarized above, this disclosure provides a class of compounds comprising a ligand moiety that specifically binds to the cell surface mannose-6-phosphate receptor (M6PR). The M6PR ligand moiety of this disclosure can be ligated to various parts of interest without affecting the specific binding to the cell surface receptor M6PR and its function. We have demonstrated that the compounds of this disclosure can utilize the function of cell surface M6PR in biological systems, for example, internalization and sequestration of the compounds into cellular lysosomes, and possibly subsequent lysosomal degradation. The compounds of this disclosure have been used in a variety of applications.

[0017] The compounds of this disclosure can specifically bind to cell surface M6PR, for example, to the internalized M6PR cell surface receptor. In certain embodiments, the surface M6PR is human M6PR. In certain embodiments, the M6PR is the Homo sapiens insulin-like growth factor 2 receptor (IGF2R) (see, for example, NCBI reference sequence: NM_000876.3), also known as the cation-independent mannose-6-phosphate receptor (CI-MPR). MP6R endogenously transports proteins with N-linked glycans capped with mannose-6-phosphate (M6P) residues to lysosomes, where they circulate between endosomes, the cell surface, and the Golgi complex. For example, Ghosh et al., Nat. Rev. Mol. Cell. See Biol. 2003;4:202-213.

[0018] The M6PR-binding compounds of the present disclosure include a moiety (X) that specifically binds to the cell surface receptor M6PR. For example, mannose-6-phosphate (M6P) or an M6P analog or derivative that specifically binds to cell surface M6PR (e.g., as described herein). The M6PR-binding compound can be monovalent or multivalent (e.g., divalent or trivalent or higher valency), a monovalent compound contains a single M6PR ligand moiety, and a multivalent compound contains two or more such moieties.

[0019] Compounds containing such X (e.g., as described herein) can bind to other receptors and can bind with lower affinity, e.g., as determined by an immunoassay or other assay known in the art. In certain embodiments, X, or a compound or conjugate described herein containing such X, specifically binds to cell surface M6PR with an affinity that is at least 2 logs, 2.5 logs, 3 logs, 4 logs or more greater than the affinity when X or the compound or conjugate binds to another cell surface receptor. In certain embodiments, X, e.g., M6P or an M6P analog or derivative, or a compound described herein containing X specifically binds to M6PR with an affinity (K d ) of 20 mM or less. In certain embodiments, such binding has an affinity (K d ) of about 20 mM or less, about 10 mM, about 1 mM, about 100 μM, about 10 μM, about 1 μM, about 100 nM, about 10 nM, or 1 nM or less. Unless otherwise specified, "binds", "binds to", "specifically binds", or "specifically binds to" in this context are used interchangeably.

[0020] In certain embodiments, the M6PR-binding moiety X can bind to an M6PR-specific cell surface receptor and direct (or target) the molecule to this receptor. In certain embodiments, the M6PR-binding moiety X can bind to M6PR and direct (or target) the compounds or conjugates described herein for internalization and sequestration into lysosomes, and / or subsequent lysosomal degradation.

[0021] In some embodiments, the M6PR bond X comprises a mannose sugar ring or an analogue thereof having a hydrophilic head group linked to the 5th position of the ring via a linking portion. The linking portion may be 1 to 6 atoms in length, such as 1 to 5, 1 to 4, or 1 to 3 atoms. The hydrophilic head group may be any convenient group that is charged or readily capable of hydrogen bonding or electrostatic interaction under aqueous or physiological conditions. The hydrophilic head group may be a structural or functional mimic of the 6-phosphate group of M6P having desirable stability. The hydrophilic head group may have a MW of less than 200, e.g., less than 150 or less than 100. In some embodiments, the hydrophilic head group is a phosphonate. In some embodiments, the hydrophilic head group is a thiophosphonate. In some embodiments, the hydrophilic head group is a phosphate, thiophosphate, or dithiophosphate.

[0022] In some embodiments, the mannose sugar ring of X is linked to an optionally substituted aryl or heteroaryl group, which together provide a moiety with desirable binding affinity and activity at the M6P receptor of interest. Multiple M6PR binding moieties X can be linked together to provide a polyvalent bond to M6PR. The M6PR binding moiety or multiple moieties X can be further linked to any convenient moiety or molecule of interest (as described herein, for example).

[0023] Therefore, the M6PR bonded compound of formula (Ia): [ka] or a salt thereof provided herein During the ceremony, X is a portion that binds to cell surface M6PR (e.g., M6PR ligand or binding site, e.g., as described herein), n is between 1 and 500. L is a linker of a defined length, Y is the part of interest.

[0024] The M6PR bond (X) of the compounds disclosed herein may include a mannose ring or an analog thereof as described by the following structure: [ka] During the ceremony, W is a hydrophilic head group, Z 1 However, it is selected from arbitrarily substituted (C1-C3) alkylenes and arbitrarily substituted ethenylenes. Z 2 However, O, S, NR 21 , and C(R 22 ) Selected from 2, each R 21 However, independently, H is selected from optionally substituted (C1-C6) alkyl groups, and each R 22 However, these are independently selected from H, halogens (e.g., F), and optionally substituted (C1-C6) alkyl groups.

[0025] The mannose ring or analogue of the M6PR bonding portion is connected to Z via the connecting portion. 2 By bonding to the base, it can be incorporated into the compounds of this disclosure. In the compound of formula (Ia), Z 2 It is understood that the bonded group or linkage portion may, in some cases, be considered part of the M6PR bond portion (X) and provide the desired bond to M6PR. In other specific cases, Z 2 The bonded group or linkage can be considered part of the linker L in formula (Ia).

[0026] In one embodiment, a cell surface mannose-6-phosphate receptor (M6PR) binding compound of formula (XI): [ka] or a salt thereof provided herein During the ceremony, Each W is independently a hydrophilic head group. each Z 1 However, independently, they are selected from arbitrarily substituted (C1-C3) alkylenes and arbitrarily substituted ethenylenes. each Z 2 However, independently, O, S, NR 21 , and C(R 22 ) Selected from 2, each R 21 However, independently, H is selected from optionally substituted (C1-C6) alkyl groups, and each R 22 However, independently selected from H, halogens (e.g., F), and optionally substituted (C1-C6) alkyl groups, Each Ar is independently an arbitrarily substituted aryl or heteroaryl group or linking moiety. each Z 3 However, it is an independent connecting part, n is between 1 and 500. L is the linker, Y is the part of interest.

[0027] In some embodiments of formula (XI), when n is 1 and Ar is phenyl, then i) L contains a skeleton of at least 16 consecutive atoms (e.g., at least 20 consecutive atoms, possibly up to about 200 consecutive atoms), ii) Y is a biomolecule, and / or ii) Z 3 However, the bonded portion is an amide, sulfonamide, urea, or thiourea.

[0028] The Ar group linkage in formula (XI) can be a monocyclic aryl or monocyclic heteroaryl group. In some embodiments of formula (XI), Ar is a five-membered monocyclic heteroaryl group. In some embodiments of formula (XI), Ar is a six-membered monocyclic aryl or heteroaryl group. The Ar group linkage in formula (XI) can be a polycyclic aryl or polycyclic heteroaryl group, such as a bicyclic aryl or bicyclic heteroaryl group. In some embodiments of formula (XI), Ar is a fused bicyclic group. In some embodiments of formula (XI), Ar is a bicyclic group comprising two aryl and / or heteroaryl monocyclic rings linked via a covalent bond. In some embodiments of formula (XI), Ar is a bicyclic aryl or bicyclic heteroaryl group having two six-membered rings. In some embodiments of formula (XI), Ar is a bicyclic aryl or bicyclic heteroaryl group having one six-membered ring linked via a covalent bond or fused to a five-membered ring.

[0029] In some embodiments of formula (XI), each Ar is independently 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. In some embodiments of formula (XI), Ar is substituted with at least one OH substituent. In some embodiments of formula (XI), Ar is substituted with one, two, or more OH groups. In some embodiments of formula (XI), Ar is substituted with at least one optionally substituted (C1-C6) alkyl.

[0030] In some embodiments of formula (XI), Ar is optionally substituted with 1,4-phenylene, optionally substituted with 1,3-phenylene, or optionally substituted with 2,5-pyridylene.

[0031] In some embodiments of formula (XI), the compound is a compound of formula (XIIa) or (XIIb): [ka] or a salt thereof, During the ceremony, Each R 11 ~R 14 However, independently, 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 Selected from, Each R 25 However, these are independently selected from H and optionally substituted (C1-C6) alkyl groups.

[0032] In some embodiments of equations (XIIa) to (XIIb), R 11 ~R 14 Each of these is H. In some embodiments of equations (XIIa) to (XIIb), R 11 ~R 14 At least one of them is OH, for example, R 11 ~R 14 One, two, or more of these are OH groups.

[0033] In some embodiments of equations (XIIa) to (XIIb), 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 -, -NR 23 SO2- and -SO2NR 23 - Select from, X 1 and X 2However, O, S, and NR 23 Selected from, R 23 and R 24 However, independently, H, C (1-3) -alkyl (e.g., methyl), and substituted C (1-3) - Selected from alkyl groups.

[0034] In some embodiments of equations (XI) to (XIIb), Z 3 This is a covalent bond to L.

[0035] In some embodiments of equations (XI) to (XIIb), Z 3 is an optionally substituted amide, urea, or thiourea. In some embodiments of formulas (XI) to (XIIb), Z 3 The following is true: [ka] During the ceremony, X 1 However, it is either O or S, t is 0 or 1, Each R 23 However, independently, H, C (1-3) -Alkyl (e.g., methyl or ethyl), and substituted C (1-3) - Selected from alkyl groups. Z 3 In some embodiments of X 1 It is O. 3 In some embodiments of X 1 S is. Z 3 In some embodiments, t is 0 and X 1 is O, and therefore Z 3 Z is an amide. 3 In some embodiments, t is 1, and therefore Z 3 It is urea or thiourea.

[0036] In some embodiments of equations (XI) to (XIIb), Z 3 is -N(R 23)SO2- or -SO2N(R 23 )-.

[0037] In some embodiments of formulas (XI) to (XIIb), Z 3 is -N(R 23 )CO- or -CON(R 23 )-.

[0038] In some embodiments of formulas (XI) to (XIIb), Z 3 is -NHC(=X 1 )NH-, and X 1 is O or S. In some embodiments, X 1 is O. In some embodiments, X 1 is S.

[0039] In some embodiments of formulas (XI) to (XIIb), -Ar-Z 3 - is selected from the following.

Chemical formula

[0040] In some embodiments of formulas (XI) to (XIIb), Z 3 is an optionally substituted triazole. When Z 3 is an optionally substituted triazole, it can be synthetically derived from the click chemical conjugation of an azide-containing precursor and an alkyne-containing precursor of the compound. Thus, in some embodiments of formulas (XIIa) to (XIIb), the compound is a compound of formula (XIIc) or (XIId):

Chemical formula

[0041] 11 ~R 14 are each H. In some embodiments of formulas (XIIc)-(XIId), at least one of R 11 ~R 14 is OH, for example, one, two or more of R 11 ~R 14 3 are OH.

[0042] In some embodiments of formulas (XIIc)-(XIId), -Ar-Z 3 - is selected from the following.

Chemical formula

[0043] In some embodiments of formula (XI), Ar is optionally substituted fused bicyclic aryl or heteroaryl. In some embodiments of formula (XI), Ar is optionally substituted naphthalene or optionally substituted quinoline. In some embodiments of formula (XI), the compound is a compound of formula (XIIIa), (XIIIb), or (XIIIb’):

Chemical formula

[0044] In some embodiments of formulas (XIIIa) to (XIIIb'), the compound is the compound of formulas (XIIIc) to (XIIIh): [ka] or its salt.

[0045] In some embodiments of equations (XIIIa) to (XIIIh), R 11 ~R 14 In some embodiments of equations (XIIIa) to (XIIIh), R 11 ~R 15 At least one of them is OH, for example, R 11 ~R 15 One, two, or more of these are OH groups.

[0046] In some embodiments of equations (XIIIa) to (XIIIh), 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 =NX2 , -N(R 23 )SO2-, and -SO2N(R 23 )- Selected from, X 1 and X 2 O, S, and NR 23 Selected from, R 23 and R 24 H and C are independent of each other. (1-3) -alkyl (e.g., methyl), and substituted C (1-3) - Selected from alkyl groups.

[0047] In some embodiments of equations (XIIIa) to (XIIIh), Z 3 This is a covalent bond to L.

[0048] In some embodiments of equations (XIIIa) to (XIIIh), Z 3 is an optionally substituted amide, urea, or thiourea. In some embodiments of formulas (XIIIa) to (XIIIh), Z 3 The following is true: [ka] During the ceremony, X 1 However, it is either O or S, t is 0 or 1, Each R 23 However, independently, H, C (1-3) -Alkyl (e.g., methyl or ethyl), and substituted C (1-3) - Selected from alkyl groups. Z 3 In some embodiments of X 1 It is O. 3 In some embodiments of X 1 S is Z 3 In some embodiments, t is 0 and X 1 is O, and therefore Z 3 Z is an amide. 3 In some embodiments, t is 1, and therefore Z 3 It is urea or thiourea.

[0049] In some embodiments of equations (XIIIa) to (XIIIh), Z 3 is -N(R 23 )SO2- or -SO2N(R 23 )-is.

[0050] In some embodiments of equations (XIIIa) to (XIIIh), Z 3 is -N(R 23 )CO- or -CON(R 23 )-is.

[0051] In some embodiments of equations (XIIIa) to (XIIIh), Z 3 -NHC(=X 1 )NH- and X 1 is O or S. In some embodiments, X 1 In some embodiments, X 1 S is.

[0052] In some embodiments of equations (XIIIa) to (XIIIh), Z 3 This is a triazole with arbitrary substitutions. 3 However, if it is an optionally substituted triazole, it can be synthetically derived from click chemical conjugation of azide-containing and alkyne-containing precursors of the compound.

[0053] In some embodiments of equations (XIIIa) to (XIIIh), -Ar-Z 3 - is selected from the following: [ka] [ka]

[0054] In some embodiments of formula (XI), Ar is an optionally substituted bicyclic aryl or optionally substituted bicyclic heteroaryl, and the compound is a compound of formula (XIVa), [ka] or a salt thereof, During the ceremony, Each Cy is independently a monocyclic aryl or monocyclic heteroaryl. Each R 11 ~R 15 However, independently, 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 Selected from, s is between 0 and 4, Each R 25 However, these are independently selected from H and optionally substituted (C1-C6) alkyl groups.

[0055] In some embodiments of formula (XIVa), Ar is an optionally substituted biphenyl, Cy is an optionally substituted phenyl, and the compound is the compound of formula (XIVb): [ka] or its salt.

[0056] In some embodiments of formula (XIVb), the compound is a compound of formula (XIVc) or (XIVd): [ka] or its salt.

[0057] In some embodiments of formulas (XI) to (XIVd), Ar is substituted with at least one OH substituent. In some embodiments of formulas (XI) to (XIVd), R 11 ~R 15 Each of these is H. In some embodiments of equations (XI) to (XIVd), R 11 ~R 15 At least one of them is OH, for example, R 11 ~R 15 One, two, or more of these are OH groups.

[0058] In some embodiments of equations (XI) to (XIVd), 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 )- Selected from, X 1 and X 2 O, S, and NR 23 Selected from, R 23 and R 24 H and C are independent of each other. (1-3) -alkyl (e.g., methyl), and substituted C (1-3) - Selected from alkyl groups.

[0059] In some embodiments of equations (XI) to (XIVd), Z 3 This is a covalent bond to L.

[0060] In some embodiments of equations (XI) to (XIVd), Z 3 is an optionally substituted amide, urea, or thiourea. In some embodiments of formulas (XI) to (XIVd), Z3 The following is true: [ka] During the ceremony, X 1 However, it is either O or S, t is 0 or 1, Each R 23 However, independently, H, C (1-3) -Alkyl (e.g., methyl or ethyl), and substituted C (1-3) - Selected from alkyl groups. Z 3 In some embodiments of X 1 It is O. 3 In some embodiments of X 1 S is Z 3 In some embodiments, t is 0 and X 1 is O, and therefore Z 3 Z is an amide. 3 In some embodiments, t is 1, and therefore Z 3 It is urea or thiourea.

[0061] In some embodiments of equations (XI) to (XIVd), Z 3 -NHC(=X 1 )NH- and X 1 is O or S. In some embodiments, X 1 In some embodiments, X 1 S is.

[0062] In some embodiments of equations (XI) to (XIVd), Z 3 is -N(R 23 )SO2- or -SO2N(R 23 )-is.

[0063] In some embodiments of equations (XI) to (XIVd), Z 3 This is a triazole with arbitrary substitutions. 3However, if it is an optionally substituted triazole, it can be synthetically derived from click chemical conjugation of azide-containing and alkyne-containing precursors of the compound.

[0064] In some embodiments of equations (XI) to (XIVd), -Ar-Z 3 - is selected from the following: [ka]

[0065] In some embodiments of formula (XI), Ar is an optionally substituted monocyclic heteroaryl. In some embodiments of formula (XI), Ar is a triazole, and the compound is a compound of formula (XVa) or (XVb). [ka]

[0066] In some embodiments of formula (XVa) or (XVb), Z 2 is O or S. In some embodiments of formula (XVa) or (XVb), Z 2 This is CH2.

[0067] In some embodiments of equations (XI) to (XVb), n is at least 2, L is a branched linker that covalently bonds each Ar group to Y. In some embodiments of formulas (XI) to (XVb), n is 2 to 20, for example n is 2 to 10, 2 to 6, for example 2 or 3.

[0068] In some embodiments of formulas (XI) to (XVb), n is 20 to 500 (e.g., 20 to 400, 20 to 300, or 20 to 200, or 50 to 500, or 100 to 500), L is an α-amino acid polymer (e.g., poly-L-lysine), and numerous -Ar-Z 3- The group is covalently bonded to the polymer backbone via a side chain group (for example, via conjugation of a lysine residue to a side chain amino group).

[0069] In some embodiments of equations (XI) to (XVb), n is at least 2, and each Z 3 The linkage is through linker L by at least 16 consecutive atomic chains (for example, by at least 20, at least 25, or at least 30 consecutive atomic chains, and optionally by up to 100 consecutive atomic chains), and all other Z 3 It is separated from the joint.

[0070] In some embodiments of formulas (XI) to (XVb), the compound is the compound of formula (XVI): [ka] or a salt thereof, During the ceremony, n is between 1 and 500. Each L 1 ~L 7 However, independently, n Z 2 A linking portion that provides a linear or branched linker together between the base and Y, -(L 1 ) a - comprises a linking moiety Ar which is an optionally substituted aryl or heteroaryl group, a is either 1 or 2, b, c, d, e, f, and g are each independently 0, 1, or 2.

[0071] In some embodiments of formula (XVI), the linear or branched linker is formed by a chain of at least 16 consecutive atoms (e.g., at least 20 consecutive atoms, at least 30 consecutive atoms, or 16 to 100 consecutive atoms) in each Z 2 And separate Y.

[0072] In some embodiments of formula (XVI), n is 1 to 20, for example, 1 to 10, 1 to 6, or 1 to 5. In some embodiments of formula (XVI), n is at least 2, for example, n is 2 or 3. In some embodiments of formula (XVI), if d is > 0, L 4 This is a branched connection point, and each L 1 The connecting parts are covalently bonded.

[0073] In some embodiments of formula (XVI), the compound is the compound of formula (XVIa), [ka] (XVIa) During the ceremony, Ar is an optionally substituted aryl or heteroaryl group. Z 11 However, it is a connecting part, r is 0 or 1, n is between 1 and 6.

[0074] In some embodiments of equation (XVIa), Z 11 However, these are covalent bonds, heteroatoms, groups with a skeleton of 1 to 3 atoms in length (e.g., -NH-, urea, thiourea, ether, amide), or triazoles.

[0075] In some embodiments of formula (XVIa), Ar is a monocyclic aryl or heteroaryl group. In some embodiments of formula (XVIa), Ar is a bicyclic aryl or heteroaryl group. In some embodiments of formula (XVIa), Ar is a tricyclic aryl or heteroaryl group. In some embodiments of formula (XVIa), Ar is selected from optionally substituted phenyl, optionally substituted biphenyl, optionally substituted naphthalene, optionally substituted triazole, optionally substituted phenyl-triazole, optionally substituted biphenyl-triazole, and optionally substituted naphthalene-triazole. In a particular embodiment, Ar is optionally substituted 1,4-phenylene.

[0076] In some embodiments of formula (XVIa), Ar is substituted with at least one hydroxyl group.

[0077] In some embodiments of equations (XVI) to (XVIa), L 1 or -Ar-(Z 11 ) r -teeth, [ka] , selected from, During the ceremony, Cy is a monocyclic aryl or heteroaryl compound. r is 0 or 1, s is between 0 and 4 (for example, 0 to 3, or 0, 1, or 2), R 11 ~R 14 and each R 15 However, independently, 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 Selected from, each R 25However, independently, H, C (1-6) -alkyl and substituted C (1-6) - Selected from alkyl groups, Z 11 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 -, and selected from optionally substituted triazoles, X 1 and X 2 However, O, S, and NR 23 Selected from, R 23 and R 24 However, independently, H, C (1-3) -alkyl (e.g., methyl), and substituted C (1-3) - Selected from alkyl groups.

[0078] In some embodiments, r is 0, Z 11 However, it does not exist. In some embodiments, r is 1.

[0079] In some embodiments of equations (XVI) to (XVIa), L 1 or -Ar-(Z 11 ) r - is as follows: [ka] In some embodiments, r is 0, Z 11 However, it does not exist. In some embodiments, r is 1.

[0080] In some embodiments of equations (XVI) to (XVIa), L 1 or -Ar-(Z 11 ) r - is as follows: [ka] In some embodiments, r is 0, Z 11 However, it does not exist. In some embodiments, r is 1.

[0081] In some embodiments of equations (XVI) to (XVIa), L 1 or -Ar-(Z 11 ) r - is as follows: [ka] In some embodiments, r is 0, Z 11 However, it does not exist. In some embodiments, r is 1.

[0082] In some embodiments of equations (XVI) to (XVIa), L 1 or -Ar-(Z 11 ) r - is as follows: [ka] In some embodiments, r is 0, Z 11 However, it does not exist. In some embodiments, r is 1.

[0083] In some embodiments of equations (XVI) to (XVIa), L 1 or -Ar-(Z 11 ) r - is as follows: [ka]

[0084] In some embodiments, r is 0, Z 11 However, it does not exist. In some embodiments, r is 1, and Z 11 However, -O-, -NR 23 -, -NR 23 CO-, CONR23 -, -NR 23 CO2-, -OCONR 23 -, -NR 23 C(=X 1 )NR 23 -, -CR 24 =N-, -CR 24 =NX 2 -, -NR 23 SO2- and -SO2NR 23 - Select from, X 1 and X 2 However, O, S and NR 23 Selected from, each R 23 and R 24 However, independently, H, C (1-3) -alkyl (e.g., methyl), and substituted C (1-3) - Selected from alkyl groups.

[0085] In some embodiments, r is 1, Z 11 The following is true: [ka] During the ceremony, X 1 However, it is either O or S, t is 0 or 1, Each R 23 However, independently, H, C (1-3) -alkyl (e.g., methyl), and substituted C (1-3) -Selected from alkyl. In some embodiments, Z 11 -NHC(=X 1 )NH- and X 1 is O or S. In some embodiments, r is 1 and Z 11 It is a triazole.

[0086] In some embodiments of equations (XI) to (XVIa), Z 3 is -N(R 23 )SO2- or -SO2N(R 23 )-is.

[0087] In some embodiments of equations (XI) to (XVIa), Z 3 is -N(R 23 )CO- or -CON(R 23 )-is.

[0088] In some embodiments of formulas (XI) to (XVIa), the hydrophilic head group W is charged, for example, can form salts under aqueous or physiological conditions. In some embodiments of formulas (XI) to (XVIa), the hydrophilic head group W is neutral.

[0089] In any one of the embodiments of formulas (XI) to (XVIa) described herein, the hydrophilic head group W is -OH, -CR 2 R 2 OH, -OP=O(OH)2, -SP=O(OH)2, -NR 3 P=O(OH)2, -OP=O(SH)(OH), -SP=O(SH)(OH), -OP=S(OH)2, -OP=O(N(R 3 )2)(OH), -OP=O(R 3 )(OH), -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 R 4 -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 -SO2NH2, -SO2NHR 3 -SO2NR 3 R 4 , -SO2NHCOR3 , -NHCOR 3 -NHC(O)CO2H, -NHSO2NHR 3 ,-NHC(O)NHS(O)2R 3 , -NHSO2R 3 -NHSO3H, [ka] , or their salts, selected from During the ceremony, R 1 and R 2 However, independently, hydrogen, SR 3 , halo, or CN, R 3 and R 4 However, independently, H, C 1-6 Alkyl or substituted C 1-6 It is an alkyl group (e.g., -CF3 or -CH2CF3), A, B, and C are each independently CH or N. Each D is independently either O or S.

[0090] In some embodiments of formulas (XI) to (XVIa), the hydrophilic head group W is a phosphate or thiophosphate (e.g., -OP=O(OH)2, -SP=O(OH)2, -OP=O(SH)(OH), -SP=O(SH)(OH), or -OP=S(OH)2). In some embodiments of formulas (XI) to (XVIa), 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 (XI) to (XVIa), the hydrophilic head group W is a sulfonate (e.g., -SO3H or a salt thereof). In some embodiments of formulas (XI) to (XVIa), the hydrophilic head group W is -CO2H or a salt thereof. In some embodiments of formulas (XI) to (XVIa), the hydrophilic head group W is a malonate (e.g., -CH(COOH)2 or a salt thereof).

[0091] In some embodiments of formulas (XI) to (XVIa), the hydrophilic head group W is a five-membered heterocycle as follows: [ka] , or including their salts.

[0092] Exemplary hydrophilic head groups W are shown in the X group of Table 1 and in the compounds of Tables 5-7B.

[0093] In some embodiments of formulas (XI) to (XVIa), a connecting portion (Z) connects the hydrophilic head group W to the mannose ring. 1 ) is -(CH2) j - and j is 1 to 3. In some embodiments, j is 2. In some embodiments of formulas (XI) to (XVIa), the connecting portion (Z) connects the hydrophilic head group W to the mannose ring. 1 ) is -CH=CH-.

[0094] In some embodiments of formulas (XI) to (XVIa), a connecting portion (Z) connects the mannose ring to the Ar group. 2 ) is either O or S. In some embodiments of equations (XI) to (XVIa), Z 2 -NR 21 - and R 21 is selected from H, optionally substituted (C1-C6) alkyl groups. In some embodiments of formulas (XI) to (XVIa), Z 2 is -NH-. In some embodiments of equations (XI) to (XVIa), Z 2 -C(R 22 )2-, and each R 22 Z is independently selected from H, a halogen (e.g., F), and optionally substituted (C1-C6) alkyl groups. In some embodiments of formulas (XI) to (XVIa), Z 2 is CH2. In some embodiments of equations (XI) to (XVIa), Z 2 This is -CF2- or -C(CH3)2-.

[0095] In some embodiments of equations (XI) to (XVIa), Z 1 is, -(CH2) j - and -CH=CH- are selected, and j is 1 to 3, Z 2 It is selected from O and CH2.

[0096] In some embodiments of equations (XI) to (XVIa), Z 1 is, -(CH2) j - and j is 2, Z 2 It is O.

[0097] In some embodiments of equations (XI) to (XVIa), Z 1 is, -(CH2) j - and j is 2, Z 2 This is CH2.

[0098] In some embodiments of equations (XI) to (XVIa), Z 1 -CH=CH- and Z 2 It is O.

[0099] In some embodiments of equations (XI) to (XVIa), Z 1 -CH=CH- and Z 2 This is CH2.

[0100] As summarized above, the M6PR bond (X) of the compounds of this disclosure (e.g., of formula (Ia)) may include a mannose ring or an analogue thereof as described by the following structure: [ka] During the ceremony, W is a hydrophilic head group, Z 1 However, it is selected from arbitrarily substituted (C1-C3) alkylenes and arbitrarily substituted ethenylenes. Z 2 However, O, S, NR 21 , and C(R 22 ) Selected from 2, each R21 However, independently, H is selected from optionally substituted (C1-C6) alkyl groups, and each R 22 However, these are independently selected from H, halogens (e.g., F), and optionally substituted (C1-C6) alkyl groups.

[0101] The mannose ring or analogue of the M6PR bonding portion is connected to Z via the connecting portion. 2 By bonding to the base, it can be incorporated into the compounds of this disclosure. In the compound of formula (Ia), Z 2 It is understood that the bonded group or linkage may, in some cases, be considered part of the M6PR linkage (X) and provide the desired bond to M6PR. For example, see formulas (XI) to (XVIa), where the aryl or heteroaryl linkage is Z 2 It is bonded to the mannose ring or analogue via a group. In other specific cases, Z 2 The bonded group or linkage can be considered part of the linker L in formula (Ia).

[0102] In some embodiments of the M6PR-binding compounds of this disclosure, for example, the compound of formula (Ia), the M6PR-binding moiety X includes the group of formulas (IIIa), (IIIb), (IIIc), or (IIId): [ka] In the formula, R'' (e.g., hydrophilic head group) is -OH, -CR 1 R 2 OH, -P=O(OH)2, P(=O)R 1 OH, -PH(=O)OH, -(CR 1 R 2 )-P=O(OH)2, -SO2OH, -S(O)OH, -OSO2OH, -COOH, -CONH2, -CONHR 3 ,-CONR 3 R 4 -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 -SO2NH2, -SO2NHR 3 -SO2NR 3 R 4 , -SO2NHCOR 3 , -NHCOR 3 ,-NHC(O)NHS(O)2R 3 , -NHSO2R 3 , [ka] Selected from the group consisting of, j is an integer between 1 and 3. R 1 and R 2 However, each is independently hydrogen, halo, or CN. R 3 and R 4 However, each is independent of C 1-6 It is alkyl, A, B, and C are each independently CH or N, and D is each independently O or S.

[0103] In some embodiments of formulas (IIIa), (IIIb), (IIIc), or (IIId), R'' is -OH, -CR 1 R 2 OH, -P=O(OH)2, P(=O)R 1 OH, -(CR 1 R 2 )-P=O(OH)2, -SO2OH, -OSO2OH, -COOH, -CONH2, -CONHR 1 ,-CONR 3 R 4 -CONHSO2R 3 -CH(COOH)2, -CR 1 R 2 COOH, -SO2R 3 -SOR 3 R 4 -SO2NH2, -SO2NHR 3 -SO2NR3 R 4 , -NHCOR 3 , -NHSO2R 3 , [ka] Selected from the group consisting of, j is an integer between 1 and 3. R 1 and R 2 However, each is independently hydrogen, halo, or CN. R 3 and R 4 However, each is independent of C 1-6 It is alkyl, A, B, and C are each independently CH or N.

[0104] In a particular embodiment, X is a group of formulas (IIIa'), (IIIa''), (IIIb'), (IIIb''), (IIIc'), (IIIc''), (IIId'), or (IIId''): [ka] R'' is as defined herein, and j is an integer between 1 and 3.

[0105] In a particular embodiment, X is X of formula (IIIa'), (IIIa''), (IIIb'), or (IIIb''). In a particular embodiment, X is X of formula (IIIc'), (IIIc''), (IIId'), or (IIId''). In a particular embodiment, X is X of formula (IIIa') or (IIIa''). In a particular embodiment, X is X of formula (IIIb') or (IIIb''). In a particular embodiment, X is X of formula (IIIc') or (IIIc''). In a particular embodiment, X is X of formula (IIId') or (IIId''). In a particular embodiment, X is X of formula (IIIa'). In one embodiment, X is X of formula (IIIa''). In a particular embodiment, X is X of formula (IIIb'). In one embodiment, X is X of formula (IIIb''). In a particular embodiment, X is X in formula (IIIc'). In one embodiment, X is X in formula (IIIc''). In a particular embodiment, X is X in formula (IIId'). In one embodiment, X is X in formula (IIId''). In a particular embodiment, X is X in formula (IIIe).

[0106] In one embodiment, j is 1 or 2. In another embodiment, j is 2 or 3. In yet another embodiment, j is 1. In yet another embodiment, j is 2. In yet another embodiment, j is 3.

[0107] In certain embodiments, R'' is -OH, -CR 1 R 2 OH, -P=O(OH)2, P(=O)R 1 OH, -(CR 1 R 2 )-P=O(OH)2, -SO2OH, -OSO2OH, -COOH, -CONH2, -CONHR 1 ,-CONR 3 R 4 -CONHSO2R 3 -CH(COOH)2, -CR 1 R2 COOH, -SO2R 3 -SOR 3 R 4 -SO2NH2, -SO2NHR 3 -SO2NR 3 R 4 , -NHCOR 3 , -NHSO2R 3 , [ka] Selected from the group consisting of, R 1 and R 2 However, each is independently hydrogen, halo, or CN. R 3 and R 4 However, each is independent of C 1-6 It is alkyl, A, B, and C are each independently CH or N. In a particular embodiment, R'' is not OH.

[0108] In certain embodiments, R'' is -OH, -CR 1 R 2 OH, -P(=O)R 1 OH, -(CR 1 R 2 )-P=O(OH)2, -SO2OH, -OSO2OH, -COOH, -CONH2, -CONHR 1 ,-CONR 3 R 4 -CONHSO2R 3 -CH(COOH)2, -CR 1 R 2 COOH, -SO2R 3 -SOR 3 R 4 -SO2NH2, -SO2NHR 3 -SO2NR 3 R 4 , -NHCOR 3 , -NHSO2R 3 , [ka] It is selected from the group consisting of the following.

[0109] In a particular embodiment, R'' is -CR 1 R 2 OH, -P(=O)R 1 OH, -(CR 1 R 2 )-P=O(OH)2, -SO2OH, -OSO2OH, -COOH, -CONH2, -CONHR 1 ,-CONR 3 R 4 -CONHSO2R 3 -CH(COOH)2, -CR 1 R 2 COOH, -SO2R 3 -SOR 3 R 4 -SO2NH2, -SO2NHR 3 -SO2NR 3 R 4 , -NHCOR 3 , -NHSO2R 3 , [ka] It is selected from the group consisting of the following.

[0110] In a particular embodiment, R'' is -P=O(OH)2, P(=O)R 1 OH, and -(CR 1 R 2 ) is selected from the group consisting of -P=O(OH)2. In a particular embodiment, R'' is -SO2OH, -OSO2OH, -CONHSO2R 3 , -SO2R 3 -SOR 3 R 4 -SO2NH2, -SO2NHR 3 -SO2NR 3 R 4 , and -NHSO2R 3 Selected from the group consisting of -OH or -CR. In a particular embodiment, R'' is -OH or -CR 1 R 2It is OH. In certain embodiments, R'' is -COOH, -CONH2, -CONHR 1 ,-CONR 3 R 4 -CH(COOH)2, -CR 1 R 2 COOH and -NHCOR 3 It is selected from the group consisting of the following.

[0111] In a particular embodiment of formula (Ia), X includes the group of formulas (IIIa-1) or (IIIb-1): [ka] During the ceremony, R L However, it is -O-, -NH-, or -CH2-, R'' is -OH, -CR 1 R 2 OH, -P=O(OH)2, P(=O)R 1 OH, -PH(=O)OH, -(CR 1 R 2 )-P=O(OH)2, -SO2OH, -S(O)OH, -OSO2OH, -COOH, -CONH2, -CONHR 3 ,-CONR 3 R 4 -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 -SO2NH2, -SO2NHR 3 -SO2NR 3 R 4 , -SO2NHCOR 3 , -NHCOR 3 ,-NHC(O)NHS(O)2R 3 , -NHSO2R 3 , [ka] Selected from the group consisting of, j is an integer between 1 and 3. R 1 and R 2 However, each is independently hydrogen, halo, or CN. R 3 and R 4 However, each is independent of C 1-6 It is alkyl, A, B, and C are each independently CH or N. Each D is independently either O or S.

[0112] In a particular embodiment of formula (Ia), X is X of formula (IIIa-1) or (IIIb-1), and R L However, if it is --O-, then R'' is [ka] Therefore, B and C are N, and j is 2.

[0113] In a particular embodiment of formula (Ia), X is X of formula (IIIa-1) or (IIIb-1), and R L is -O- and R'' is -CR 1 R 2 If it is COOH, R 1 and R 2 Neither of them is hydrogen.

[0114] In a particular embodiment of formula (Ia), X is X of formula (IIIa-1) or (IIIb-1), and R L is --O-, and R'' is, [ka] If B and C are N, then j is 2 and R L is -O- and R'' is -CR 1 R 2 If it is COOH, then R 1 and R2 Neither of them is hydrogen.

[0115] In a particular embodiment of formula (Ia), X is X of formula (IIIa-1) or (IIIb-1), and R L is -NH- or -CH2-, and R'' and the remaining variables are as described for formula (Ia).

[0116] In a particular embodiment of formula (Ia), X is X of formula (IIIa-1) or (IIIb-1), where R' is -O- and R'' is [ka] If B and C are N, then j is 2, but R' is -O- and R'' is -CR 1 R 2 If it is COOH, then R 1 and R 2 Neither of them is hydrogen.

[0117] In certain embodiments, a compound of formula (Ia) is provided herein, where X is X of formula (IIIa-1) or (IIIb-1), R' is -O-, -NH-, or -CH2-, and R'' is -P=O(OH)2, P(=O)R 1 OH, -PH(=O)OH, -(CR 1 R 2 )-P=O(OH)2, -S(O)OH, -OSO2OH, -CONH(OH), -CONH(OR 3 )-CONHSO2R 3 -CONHSO2NR 3 R 4 , -SO2R 3 -SOR 3 R 4 -SO2NH2, -SO2NHR 3 -SO2NR 3 R 4 , -SO2NHCOR 3 , -NHCOR 3 ,-NHC(O)NHS(O)2R 3 , -NHSO2R3 , [ka] The variables are selected from the group consisting of the following, and the remaining variables are as described for equation (Ia).

[0118] Table 1 shows exemplary moieties (X1-X27) that bind to M6PR, and synthons that may be used to prepare the compounds of this disclosure containing the M6PR ligand of interest. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] [Table 1-8] [Table 1-9]

[0119] ASGPR binding compounds As summarized above, this disclosure provides a class of compounds comprising a ligand moiety that specifically binds to cell surface asialoclycoprotein receptors (ASGPRs).

[0120] The term "asialoclycoprotein receptor" (ASGPR), also known as the "Ashwell Morell receptor," refers to a transmembrane glycoprotein receptor, primarily found in hepatocytes, that plays a crucial role in serum glycoproteins by mediating endocytosis and lysosomal degradation using exposed terminal galactose or N-acetylgalactosamine (GalNAc) residues. ASGPRs circulate between endosomes and the cell surface. In certain embodiments, ASGPR is Homo sapiens asialoclycoprotein receptor 1 (ASGR1) (see, for example, NCBI reference sequence: NM_001197216).

[0121] Therefore, the ASGPR-binding compound of formula (Ib): [ka] or a salt thereof provided herein During the ceremony, X is a portion that binds to a cell surface ASGPR (e.g., an ASGPR ligand or binding site, e.g., as described herein), n is between 1 and 500. L is a linker of a defined length, Y is the part of interest.

[0122] The ASGPR-binding moiety (X) of the compounds and conjugates of this disclosure may be N-acetylgalactosamine (GalNAc), or an analog or derivative of GalNAc. Various ligands capable of binding to ASGPR can be adapted for use in the compounds and conjugates of this disclosure.

[0123] In a particular embodiment, each X is independently selected from the group consisting of formulas (IIIj), (IIIk), (IIIl), and (IIIm): [ka] , R 1 However, -OH, -OC(O)R, or [ka] And R is C 1-6 It is alkyl, R 2 However, -NHCOCH3, -NHCOCF3, -NHCOCH2CF3, -OH, and [ka] Selected from the group consisting of, R 3 However, it is selected from the group consisting of -H, -OH, -CH3, -OCH3, and -OCH2CH=CH2.

[0124] In a particular embodiment, X is X in equation (IIIo). [ka]

[0125] In a particular embodiment, X is X in the formula. [ka]

[0126] In a particular embodiment, X is X in equation (IIIp). [ka]

[0127] In a particular embodiment, X is X in equation (IIIo). [ka]

[0128] In a particular embodiment, X is X in the formula. [ka]

[0129] In a particular embodiment, X is selected from the group consisting of formulas (IIIj'), (IIIk'), (IIIl'), and (IIIm'): [ka] R 1 However, -OH, -OC(O)R, or [ka] And R is C 1-6 It is alkyl, R 2 However, -NHCOCH3, -NHCOCF3, -NHCOCH2CF3, -OH, and [ka] Selected from the group consisting of, R 3 However, it is selected from the group consisting of -H, -OH, -CH3, -OCH3, and -OCH2CH=CH2.

[0130] In a particular embodiment, X is X in equation (IIIo'). [ka]

[0131] In a particular embodiment, X is X in equation (IIIp'). [ka]

[0132] In certain embodiments of the compounds described herein, each X is independently selected from the group consisting of formulas (IIIa), (IIIb), (IIIc), (IIId), (IIIe), (IIIj), (IIIk), (IIIl), (IIIm), (IIIp), (IIIj'), (IIIk'), (IIIl'), (IIIm'), and (IIIp').

[0133] In one embodiment, the compound of formula (Ib) is selected from the compounds in Table 8. In another embodiment, the compound of formula (Ib) is selected from the compounds in Table 9.

[0134] Examples of ASGPR-binding compounds of formula (Ib) are shown in Tables 8-9.

[0135] Linker The terms “linker,” “linking portion,” and “linking group” are used interchangeably and refer to a linking portion that covalently bonds two or more parts or compounds, such as a ligand and other parts of interest. In some cases, a linker is divalent and connects two parts. In some cases, a linker is a branched linking group, which is a trivalent or higher polyvalent linker. In some cases, a linker connecting two or more parts has a linear or branched skeleton with a length, measured between the two or more parts, 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, 60 atoms or less, 50 atoms or less, 40 atoms or less, 30 atoms or less, or even 20 atoms or less). The linking portion can be a covalent bond connecting two groups, or a linear or branched chain of carbon atoms with a length of 1 to 500 atoms, for example, approximately 1, 2, 3, 4, 5, 6, 8, 10, 12, 14, 16, 18, 20, 30, 40, 50, 100, 150, 200, 300, 400, or 500 atoms, and the linker can be linear, branched, cyclic, or a single atom. In some cases, one, two, three, four, five or more, ten or more, or more carbon atoms in the linker skeleton can be optionally substituted with heteroatoms, such as sulfur, nitrogen, or oxygen heteroatoms. In some cases, if the linker contains a PEG group, every third atom in that segment of the linker skeleton is substituted with oxygen. Bonds between skeleton atoms can be saturated or unsaturated, and typically there are one, two, or three or fewer unsaturated bonds in the linker skeleton. The linker may contain one or more substituents, such as alkyl, aryl, or alkenyl groups. The linker may contain, but is not limited to, one or more of the following: oligo(ethylene glycol), ether, thioether, disulfide, amide, carbonate, carbamate, tertiary amine, alkyl (which may be linear or branched), such as methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl). The linker skeleton may contain cyclic groups, such as aryl, heterocyclic, cycloalkyl, or heterocyclic groups, with two or more atoms of the cyclic group, such as two, three, or four atoms, included in the skeleton.

[0136] In some embodiments, the “linker” or linking portion originates from a molecule having two reactive ends, one for conjugation to a portion of interest (Y), such as a biomolecule (e.g., an antibody), and the other for conjugation to a portion that binds to a cell surface receptor (denoted as X). For example, if the cell surface receptor is a mannose-6-phosphate receptor (M6PR), the portion may be mannose-6-phosphate or an analogue of the mannose-6-phosphate portion. If Y is a polypeptide, the polypeptide-binding reactive end of the linker may optionally be a site that can be conjugated to the polypeptide via a cysteine-thiol or lysineamine group on the polypeptide, and therefore 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.

[0137] In certain embodiments of the formulas described herein, linker L comprises one or more linear or branched carbon moieties and / or polyether (e.g., ethylene glycol) moieties (e.g., repeating units of -CH2CH2O-), and / or combinations thereof. In certain embodiments, these linkers optionally have amide linkages, urea or thiourea linkages, carbamate linkages, ester linkages, amino linkages, ether linkages, thioether linkages, sulfhydryl linkages, or other heterofunctional linkages. 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 linkages, thioether linkages, amine linkages, amide linkages, carbon-carbon linkages, carbon-nitrogen linkages, carbon-oxygen linkages, carbon-sulfur linkages, 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.

[0138] In a particular embodiment, L is approximately 10 Å to 20 Å in length. In a particular embodiment, L is approximately 15 Å to 20 Å in length. In a particular embodiment, L is approximately 15 Å in length. In a particular embodiment, L is approximately 16 Å in length. In a particular embodiment, L is approximately 17 Å in length.

[0139] In a particular embodiment, L is a linker of approximately 5 Å to approximately 500 Å. In a particular embodiment, L is approximately 10 Å to approximately 400 Å. In a particular embodiment, L is approximately 10 Å to approximately 300 Å. In a particular embodiment, L is approximately 10 Å to approximately 200 Å. In a particular embodiment, L is approximately 10 Å to approximately 100 Å. In a particular embodiment, L is approximately 10 Å to approximately 20 Å, approximately 20 Å to approximately 30 Å, approximately 30 Å to approximately 40 Å, approximately 40 Å to approximately 50 Å, approximately 50 Å to approximately 60 Å, approximately 60 Å to approximately 70 Å, approximately 70 Å to approximately 80 Å, approximately 80 Å to approximately 90 Å, or approximately 90 Å to approximately 100 Å. In certain embodiments, L is a linker of about 5 Å to about 500 Å, and L comprises an optionally substituted arylene linked to X, an optionally substituted heteroarylene linked to X, an optionally substituted heterocyclene linked to X, or an optionally substituted cycloalkylene linked to X. In certain embodiments, L is a linker of about 10 Å to about 500 Å, and L comprises an optionally substituted arylene linked to X, an optionally substituted heteroarylene linked to X, an optionally substituted heterocyclene linked to X, or an optionally substituted cycloalkylene linked to X. In certain embodiments, L is a linker of about 10 Å to about 400 Å, and L comprises an optionally substituted arylene linked to X, an optionally substituted heteroarylene linked to X, an optionally substituted heterocyclene linked to X, or an optionally substituted cycloalkylene linked to X. In a particular embodiment, L is a linker of about 10 Å to about 200 Å, and L includes an optionally substituted arylene linked to X, an optionally substituted heteroarylene linked to X, an optionally substituted heterocyclene linked to X, or an optionally substituted cycloalkylene linked to X.

[0140] In a particular embodiment, linker L separates X and Y (or Z) by chains of 4 to 500 consecutive atoms. In a particular embodiment, linker L separates X and Y (or Z) by chains of 4 to 50 consecutive atoms. In a particular embodiment, linker L separates X and Y (or Z) by chains of 6 to 50 consecutive atoms, by chains of 11 to 50 consecutive atoms, by chains of 16 to 50 consecutive atoms, by chains of 21 to 50 consecutive atoms, by chains of 26 to 50 consecutive atoms, by chains of 31 to 50 consecutive atoms, by chains of 36 to 50 consecutive atoms, by chains of 41 to 50 consecutive atoms, or by chains of 46 to 50 consecutive atoms. In a particular embodiment, linker L separates X and Y (or Z) by chains of 6 to 50 consecutive atoms. In a particular embodiment, linker L separates X and Y (or Z) by a chain of 11 to 50 consecutive atoms. In a particular embodiment, linker L separates X and Y (or Z) by a chain of 16 to 50 consecutive atoms. In a particular embodiment, linker L separates X and Y (or Z) by a chain of 21 to 50 consecutive atoms. In a particular embodiment, linker L separates X and Y (or Z) by a chain of 26 to 50 consecutive atoms. In a particular embodiment, linker L separates X and Y (or Z) by a chain of 31 to 50 consecutive atoms. In a particular embodiment, linker L separates X and Y (or Z) by a chain of 36 to 50 consecutive atoms. In a particular embodiment, linker L separates X and Y (or Z) by a chain of 41 to 50 consecutive atoms. In a particular embodiment, linker L separates X and Y (or Z) by a chain of 46 to 50 consecutive atoms.

[0141] In a particular embodiment, linker L separates X and Y (or Z) by chains of 4 or 5 consecutive atoms, 6 to 10 consecutive atoms, 11 to 15 consecutive atoms, 16 to 20 consecutive atoms, 21 to 25 consecutive atoms, 26 to 30 consecutive atoms, 31 to 35 consecutive atoms, 36 to 40 consecutive atoms, 41 to 45 consecutive atoms, or 46 to 50 consecutive atoms. .

[0142] In a particular embodiment, linker L is a chain of 5 to 500 consecutive atoms separating X and Y (or Z), and includes optionally substituted arylenes linked to X, optionally substituted heteroarylenes linked to X, optionally substituted heterocyclenes linked to X, or optionally substituted cycloalkylenes linked to X. In a particular embodiment, linker L is a chain of 7 to 500 consecutive atoms separating X and Y (or Z), and includes optionally substituted arylenes linked to X, optionally substituted heteroarylenes linked to X, optionally substituted heterocyclenes linked to X, or optionally substituted cycloalkylenes linked to X. In a particular embodiment, linker L is a chain of 10 to 500 consecutive atoms separating X and Y (or Z), and includes optionally substituted arylenes linked to X, optionally substituted heteroarylenes linked to X, optionally substituted heterocyclenes linked to X, or optionally substituted cycloalkylenes linked to X. In a particular embodiment, linker L is a chain of 15 to 400 consecutive atoms separating X and Y (or Z), and includes optionally substituted arylenes linked to X, optionally substituted heteroarylenes linked to X, optionally substituted heterocyclenes linked to X, or optionally substituted cycloalkylenes linked to X.

[0143] In a particular embodiment, linker L is a chain of 5 to 500 consecutive atoms separating X and Y (or Z), and includes optionally substituted arylenes linked to X, or optionally substituted heteroarylenes linked to X. In a particular embodiment, linker L is a chain of 7 to 500 consecutive atoms separating X and Y (or Z), and includes optionally substituted arylenes linked to X, or optionally substituted heteroarylenes linked to X. In a particular embodiment, linker L is a chain of 10 to 500 consecutive atoms separating X and Y (or Z), and includes optionally substituted arylenes linked to X, or optionally substituted heteroarylenes linked to X. In a particular embodiment, linker L is a chain of 15 to 400 consecutive atoms separating X and Y (or Z), and includes optionally substituted arylenes linked to X, or optionally substituted heteroarylenes linked to X.

[0144] In a particular embodiment, linker L is a chain of 5 to 500 consecutive atoms separating X and Y (or Z), and includes optionally substituted phenylene linked to X. In a particular embodiment, linker L is a chain of 7 to 500 consecutive atoms separating X and Y (or Z), and includes optionally substituted phenylene linked to X. In a particular embodiment, linker L is a chain of 10 to 500 consecutive atoms separating X and Y (or Z), and includes optionally substituted phenylene linked to X. In a particular embodiment, linker L is a chain of 15 to 400 consecutive atoms separating X and Y (or Z), and includes optionally substituted phenylene linked to X.

[0145] In a particular embodiment, the linker L is a chain of 16 to 400 consecutive atoms separating X and Y (or Z), and includes optionally substituted arylenes linked to X, optionally substituted heteroarylenes linked to X, optionally substituted heteroarylenes linked to X, optionally substituted heterocycloenes linked to X, or optionally substituted cycloalkylenes linked to X.

[0146] The linker is located at the Z of the M6PR joint portion (X) (for example, as described herein). 2 It is understood that it can be considered to be directly connected to the base. In some embodiments of formula (XI), the linker is Z 3 It can be considered to be directly connected to the base. Alternatively, the -Ar-Z of equation (XI) 3 -The base (for example, as described herein) is Z 2 It can be considered as part of the connecting portion that connects to Y. This disclosure includes all such arrangements of the M6PR connecting portion (X) and linker (L).

[0147] In some embodiments of equations (I) to (Ia), L is the linker of equation (IIa): [ka] During the ceremony, Each L 1 ~L 7 However, it is an independent connecting part, a is either 1 or 2, b, c, d, e, f, and g are each independently 0, 1, or 2. n is between 1 and 500.

[0148] In some embodiments of equation (IIa), n is an integer from 1 to 5 and d is 0; n is 1 and d is 1; n is an integer from 1 to 3 and d is 2; and n is an integer from 1 to 5.

[0149] In some embodiments of formula (IIa), L1 This includes optionally substituted aryl or heteroaryl groups or linking moieties, as shown in formula (XI), for example. In some embodiments of formula (IIa), L 1 This includes optionally substituted monocyclic, bicyclic, or tricyclic aryl or heteroaryl groups (for example, as described herein). In some embodiments of formula (IIa), L 1 It further includes one or more connecting parts, each independently of C (1-10) Alkyl, -O-, -S-, -NH-, -NHCO-, -CONH-, -NHC(=O)NH-, -NHC(=S)NH-, -NHCO2-, -OC(=O)NH-, -OC(=O)-, -CO2-, -(OCH2) p -, and -(OCH2CH2) p - is selected from and p is 1-20, for example 1-10, 1-6, or 1-3, for example 1 or 2.

[0150] In some embodiments of formula (IIa), each L 1 Independently, [ka] And, z and v are independently between 0 and 10, for example, between 0 and 6 or between 0 and 3, for example, 0, 1, or 2.

[0151] In a particular embodiment of formula (IIa), L 1 teeth, [ka] That is the case.

[0152] In a particular embodiment of formula (IIa), L 1 teeth, [ka] That is the case.

[0153] In a particular embodiment of formula (IIa), L 1 teeth, [ka] That is the case.

[0154] In a particular embodiment of formula (IIa), L 1 teeth, [ka] That is the case.

[0155] In a particular embodiment of formula (IIa), L 1 teeth, [ka] That is the case.

[0156] In a particular embodiment of formula (IIa), each L 2 -C 1-6 -Alkilen-, -NHCO-C 1-6 -Alkilen-, -CONH-C 1-6 -Alkylene-, -(OCH2) p -, or -(OCH2CH2) p - and p is between 1 and 20, for example, 1 to 10, 1 to 6, or 1 to 3, for example, 1 or 2.

[0157] In a particular embodiment of formula (IIa), each L 3 Independently, [ka] , or -(OCH2CH2) q - and w and u are independently 0 to 10, e.g., 1 to 10, 1 to 6, or 1 to 3, e.g., 1 or 2, and q is 1 to 20, e.g., 1 to 10, 1 to 6, or 1 to 3, e.g., 1 or 2.

[0158] In some embodiments of formula (IIa), each L4 This refers to a linear or branched connecting section.

[0159] In some embodiments of formula (IIa), L 4 This is a branched connection, for example, a trivalent connection. For example, L 4 The connecting portion may be one of the following general formulas. [ka]

[0160] In some embodiments of formula (IIa), the branched linkage portion may be of higher valence and may be represented by one of the following general formulas: [ka] , etc. Any two L 4 The bases can be directly linked or connected via any linear linking portion (for example, as described herein).

[0161] In some embodiments of formula (IIa), the branched linkage portion may include one, two, or more L4 linkage portions, each being a trivalent portion, which, when linked together, provide multiple branching points for covalent bonding of ligands, and may be described by one of the following general formulas: [ka] t is a range from 0 to 500, for example, 0 to 100, 0 to 20, or 0 to 10.

[0162] In some embodiments, branched connecting portion (for example, L 4 ) contains one or more of the following: amino acid residues (e.g., Asp, Lys, Orn, Glu), N-substituted amides (-N(-)C(=O)-), tertiary amino acids, polyols (e.g., O-substituted glycerols), etc.

[0163] In some embodiments of formula (IIa), one or more L 4 teeth, [ka] Selected from, Each x and y is independently between 1 and 20. In some cases, each x is 1, 2, or 3, for example, 2.

[0164] In some embodiments of formula (IIa), each L 4 Independently, -OCH2CH2-, [ka] , and Each x and y is independently between 1 and 10, for example, 1 and 6, or between 1 and 3, for example, 1 or 2.

[0165] In some embodiments of formula (IIa), each L 5 It is independently, -NHCO-C 1-6 -Alkilen-, -CONH-C 1-6 -Alkilen-, -C 1-6 -Alkilen-, [ka] , or -(OCH2CH2) r - and each r is independently between 1 and 20, for example, 1 to 10, 1 to 6, or 1 to 3, for example, 1 or 2.

[0166] In some embodiments of formula (IIa), each L 6 It is independently, -NHCO-C 1-6 -Alkilen-, -CONH-C 1-6 -Alkilen-, -C 1-6 -Alkylene-, or -(OCH2CH2)- s - and s is between 1 and 20, for example, 1 to 10, 1 to 6, or 1 to 3, for example, 1 or 2.

[0167] In some embodiments of formula (IIa), each L 7 It is independently, -NHCO-C 1-6 -Alkilen-, -CONH-C 1-6 -Alkilen-, -C 1-6 -Alkilen-, -(OCH2CH2) t -, or -OCH2-, where t is 1 to 20, e.g., 1 to 10, 1 to 6, or 1 to 3, e.g., 1 or 2.

[0168] In some embodiments of formula (IIa), Each L 1 However, they became independent, [ka] And, Each L 2 However, independently, -C 1-6 -Alkilen-, -NHCO-C 1-6 -Alkilen-, -CONH-C 1-6 -Alkylene-, -(OCH2) p -, or -(OCH2CH2) p -and, Each L 3 However, they became independent, [ka] , or -(OCH2CH2) q -and, Each L 4 However, independently, -OCH2CH2-, [ka] And, Each L 5 However, it became independent, -NHCO-C 1-6 -Alkilen-, -CONH-C 1-6 -Alkilen-, -C 1-6 -Alkilen-, [ka] , or -(OCH2CH2) r -and, Each L 6 However, it became independent, -NHCO-C 1-6 -Alkilen-, -CONH-C 1-6 -Alkilen-, -C 1-6 -Alkylene-, or -(OCH2CH2)- s -and, Each L 7 However, it became independent, -NHCO-C 1-6 -Alkilen-, -CONH-C 1-6 -Alkilen-, -C 1-6 -Alkilen-, -(OCH2CH2) t -, or -OCH2- p, q, r, s, and t are each independent integers between 1 and 20. a is either 1 or 2, b, c, d, e, f, and g are each independently 0, 1, or 2. u, v, w, x, y, and z are each independent integers between 1 and 10. If n is an integer between 1 and 5 and d is 0, if n is 1 and d is 1, if n is an integer between 1 and 3 and d is 2, then n is an integer between 1 and 5.

[0169] In some embodiments of formula (IIa), Each L 1 However, they became independent, [ka] , -C 1-6 -Alkilen-, -(OCH2CH2) k -, or -(OCH2CH2) k -(CH2) v -and, Each L 2 However, independently, -C 1-6 -Alkilen-, -NHCO-C 1-6 -Alkylene-, -(OCH2) p -, or -(OCH2CH2) p -and, Each L 3 However, they became independent, [ka] , or -(OCH2CH2) q -and, Each L 4 However, independently, -OCH2CH2-, [ka] And, Each L 5 However, it became independent, -NHCO-C 1-6 -Alkylene- or -(OCH2CH2) r -and, Each L 6 However, it became independent, -NHCO-C 1-6 -Alkylene- or -(OCH2CH2) s -and, Each L 7 However, it became independent, -NHCO-C 1-6 -Alkilen-, -(OCH2CH2) t -, or -OCH2- k, p, q, r, s, and t are each independently integers between 1 and 20, a is 1 or 2, b, c, d, e, f, and g are each independently 0, 1, or 2, and u, v, w, x, y, and z are each independently integers between 1 and 10. If n is an integer between 1 and 5 and d is 0, if n is 1 and d is 1, if n is an integer between 1 and 3 and d is 2, then n is an integer between 1 and 5.

[0170] In some embodiments of formula (IIa), Each L 1 However, they became independent, [ka] And, Each L 2 However, independently, -C 1-6 -Alkilen-, -NHCO-C 1-6 -Alkylene-, -(OCH2) p-, or -(OCH2CH2) p -and, Each L 3 However, they became independent, [ka] , or -(OCH2CH2) q -and, Each L 4 However, independently, -OCH2CH2-, [ka] And, Each L 5 However, it became independent, -NHCO-C 1-6 -Alkylene- or -(OCH2CH2) r -and, Each L 6 However, it became independent, -NHCO-C 1-6 -Alkylene- or -(OCH2CH2) s -and, Each L 7 However, it became independent, -NHCO-C 1-6 -Alkilen-, -(OCH2CH2) t -, or -OCH2- p, q, r, s, and t are each independently integers between 1 and 20, a is 1 or 2, b, c, d, e, f, and g are each independently 0, 1, or 2, and u, v, w, x, y, and z are each independently integers between 1 and 10. If n is an integer between 1 and 5 and d is 0, if n is 1 and d is 1, if n is an integer between 1 and 3 and d is 2, then n is an integer between 1 and 5.

[0171] In a particular embodiment of formula (IIa), a is 1. In a particular embodiment of formula (IIa), a is 1 and b, c, d, e, f, and g are 0.

[0172] In a particular embodiment of formula (IIa), at least one of b, c, e, f, and g is not 0. In a particular embodiment of formula (IIa), a, b, c, and d are 1, and e, f, and g are 0. In a particular embodiment of formula (IIa), a, b, c, d, and g are 1, and e and f are 0. In a particular embodiment of formula (IIa), a, b, d, e, and f are 1, c and g are 0, z is an integer between 2 and 10, and n is an integer between 1 and 5.

[0173] In a particular embodiment of formula (IIa), at least one of b or c is not 0, and at least one of e, f, and g is not 0. In a particular embodiment of formula (IIa), a, b, c, d, e, and f are 1, and g is 0 or 1. In a particular embodiment of formula (IIa), a, b, c, d, e, f, and g are 1.

[0174] In a particular embodiment of formula (IIa), a, b, and c are each independently 1 or 2.

[0175] In a particular embodiment, k, p, q, r, s, and t are each independently integers between 1 and 20. In a particular embodiment, k, p, q, r, s, and t are each independently integers between 1 and 10. In a particular embodiment, k, p, q, r, s, and t are each independently integers between 1 and 5. In a particular embodiment, k, p, q, r, s, and t are each independently integers between 1 and 3.

[0176] In a particular embodiment, p, q, r, s, and t are each independently integers between 1 and 20. In a particular embodiment, p, q, r, s, and t are each independently integers between 1 and 10. In a particular embodiment, p, q, r, s, and t are each independently integers between 1 and 5. In a particular embodiment, p, q, r, s, and t are each independently integers between 1 and 3.

[0177] In a particular embodiment, u, v, w, x, y, and z are each independently integers from 1 to 10. In a particular embodiment, u, v, w, x, y, and z are each independently integers from 1 to 5. In a particular embodiment, u, v, w, x, y, and z are each independently integers from 1 to 3.

[0178] In a particular embodiment of formula (IIa), n is 1. In a particular embodiment of formula (IIa), n is 2. In a particular embodiment of formula (IIa), n is 3. In a particular embodiment of formula (IIa), n is 4. In a particular embodiment of formula (IIa), n is 5.

[0179] In yet another embodiment, a compound of formula (Ia) or (IIa) is provided herein, where L is a linker of formula (IIe): [ka] , where L 1 , L 2 , L 3 , L 4 , L 5 , and n are as defined herein.

[0180] In a particular embodiment of formula (IIe), L 1 teeth, [ka] And L 3 teeth, [ka] Therefore, a is 1, b is 0, c is 1, u is 2, and the sum of v and w is 4.

[0181] In a particular embodiment of formula (IIe), L 1 teeth, [ka] And L 2 It is methylene, and L 3 teeth, [ka] Therefore, a is 1, b is 1, c is 1, u is 2, and the sum of v and w is 3.

[0182] In a particular embodiment of formula (IIe), L 1 teeth, [ka] And L 2 It is methylene, and L 3 teeth, [ka] And a is 1, b is 2, c is 1, u is 2, v is 1, and w is 1.

[0183] In a particular embodiment of formula (IIe), L 1 teeth, [ka] And L 2 It is ethylene, L 3 teeth, [ka] And a is 1, b is 1, c is 1, u is 2, v is 1, and w is 1.

[0184] In a particular embodiment of formula (IIe), L 1 teeth, [ka] And L 2 It is methylene, and L3 teeth, [ka] And a is 1, b is 2, c is 1, u is 2, v is 1, and w is 1.

[0185] In a particular embodiment of formula (IIe), L 1 teeth, [ka] And L 3 teeth, [ka] And L 5 is -(OCH2CH2) r -, and a is 1, b is 0, c is 1, d is 0, u is 2, e is 1, and f and g are 0.

[0186] In a particular embodiment of formula (IIe), L 1 teeth, [ka] And L 3 teeth, [ka] And L 5 is -(OCH2CH2) r -, a is 1, b is 0, c is 1, d is 1, u is 2, e is 1, f and g are 0, and n is 1.

[0187] In a particular embodiment of formula (IIe), L 1 teeth, [ka] And L 3 teeth, [ka] And L 5 is -(OCH2CH2) r -, a is 1, b is 0, c is 1, d is 1, u is 2, e is 1, f and g are 0, and n is 2.

[0188] In a particular embodiment of formula (IIe), L 1 teeth, [ka] And L 3 teeth, [ka] And L 5 is -(OCH2CH2) r -, a is 1, b is 0, c is 1, d is 1, u is 2, e is 1, f and g are 0, and n is 3.

[0189] In a particular embodiment of formula (IIe), L 1 teeth, [ka] And L 3 teeth, [ka] And L 5 is -(OCH2CH2) r -, a is 1, b is 0, c is 1, d is 1, u is 2, the sum of v and w is 4, and n is 1, 2, or 3.

[0190] In a particular embodiment of formula (IIe), L 1 teeth, [ka] And L 2 It is methylene, and L 3 teeth, [ka] And L 5 is -(OCH2CH2) r -, a is 1, b is 1, c is 1, u is 2, the sum of v and w is 3, and n is 1, 2, or 3.

[0191] In a particular embodiment of formula (IIe), L 3 teeth, [ka] And L 5 is -(OCH2CH2) r -, a is 1, b is 0, c is 1, u is 2, the sum of v and w is 4, and n is 1, 2, or 3.

[0192] In a particular embodiment of formula (IIe), L 2 It is methylene, and L 3 teeth, [ka] And L 5 is -(OCH2CH2) r -, a is 1, b is 1, c is 1, u is 2, the sum of v and w is 3, and n is 1, 2, or 3.

[0193] In another embodiment, the compound of formula (Ib): [ka] Or a salt thereof, a single stereoisomer, a mixture of stereoisomers, or an isotopic form thereof is provided herein. During the ceremony, X is the part that binds to the mannose-6-phosphate receptor (M6PR) on the cell surface. Y [ka] This is a part that has the structure of [this].

[0194] In another embodiment, a compound of formula (Ia) is provided herein, in which, L is the linker in equation (IIb) below: [ka] During the ceremony, L 1 but, [ka] And, L 2 However, -(OCH2CH2) p -and, L 3 However, -NHCO-C 1-6 -Alkilen-, p is an integer between 1 and 20, a is 1, and b and c are independently either 0 or 1. n is 2, [ka] This represents the connection point to X, [ka] However, L 2 This represents a connection point to [a specific location].

[0195] In some embodiments, Y is a chemoselective ligation group (e.g., an active ester, maleimide, or isothiocyanate). In some embodiments, L 1 teeth, [ka] That is the case.

[0196] In another embodiment, a compound of formula (Ia) is provided herein, where L is a linker of the following formula (IIc): [ka] During the ceremony, L 1 but, [ka] And, L 2 but, [ka] And, L 3 However, -NHCO-C 1-6 -Alkylene- or -(OCH2CH2) p -and, L 4 However, -NHCO-C 1-6 -Alkylene- or -(OCH2CH2) q -and, p and q are each independent integers between 1 and 20, a is 1, b, c, and d are each independent 0 or 1, and w and u are each independent integers between 1 and 10. [ka] This represents the connection point to X, [ka] However, L 2 This represents a connection point to where n is 2.

[0197] In some embodiments, Y is a chemoselective ligation group (e.g., an active ester, maleimide, or isothiocyanate). In some embodiments, L 1 teeth, [ka] That is the case.

[0198] In another embodiment, a compound of formula (Ia) is provided herein, where L is a linker of formula (IId): [ka] During the ceremony, L 1 but, [ka] And, L 2 but, [ka] And, L 3 but, [ka] And, L 4 However, -CH2CH2(OCH2CH2) q -and, p is an integer between 1 and 20, c is 1, a, b, and d are each independently 0 or 1, and u, v, w, and z are each independently integers between 1 and 10. [ka] However, H or L 2 Represents the connection point to, [ka] However, L 4 Represents the connection point to, n is an integer between 1 and 5.

[0199] In some embodiments of formula (IId), Y is a chemoselective ligation group.

[0200] In a particular embodiment of formula (IId), L 3 The following applies:

change

[0201] In certain embodiments of formula (IId), X is X of formula (IIIa), (IIIb), (IIIc), or (IIId), for example, as described herein. In certain embodiments of formula (IId), X is X of formula (IIIa'), (IIIa''), (IIIb'), (IIIb''), (IIIc'), (IIIc''), (IIId'), or (IIId''), for example, as described herein. In certain embodiments, X is X of formula (IIIa'), (IIIa''), (IIIb'), or (IIIb''). In certain embodiments of formula (IId), X is X of formula (IIIc'), (IIIc''), (IIId'), or (IIId''). In certain embodiments of formula (IId), X is X of formula (IIIa') or (IIIa''). In a particular embodiment of formula (IId), X is X of formula (IIIb') or (IIIb''). In a particular embodiment of formula (IId), X is X of formula (IIIc') or (IIIc''). In a particular embodiment of formula (IId), X is X of formula (IIId') or (IIId''). In a particular embodiment of formula (IId), X is X of formula (IIIa'). In one embodiment of formula (IId), X is X of formula (IIIa''). In a particular embodiment of formula (IId), X is X of formula (IIIb'). In one embodiment of formula (IId), X is X of formula (IIIb''). In a particular embodiment of formula (IId), X is X of formula (IIIc'). In one embodiment of formula (IId), X is X of formula (IIIc''). In a particular embodiment of formula (IId), X is X of formula (IIId'). In one embodiment of formula (IId), X is X of formula (IIId''). In a particular embodiment of formula (IId), X is X of formula (IIIe). In one embodiment, j is 1 or 2. In another embodiment, j is 2 or 3. In yet another embodiment, j is 1. In yet another embodiment, j is 2. In yet another embodiment, j is 3.

[0202] In certain embodiments of formula (IId), X (e.g., as described above) comprises a hydrophilic head group (e.g., R'') as described in any one of the embodiments described herein. In certain embodiments of formula (IId), X comprises -OH, -CR 1 R 2 OH, -P=O(OH)2, P(=O)R 1 OH, -(CR 1 R 2 )-P=O(OH)2, -SO2OH, -OSO2OH, -COOH, -CONH2, -CONHR 1 ,-CONR 3 R 4 -CONHSO2R 3 -CH(COOH)2, -CR 1 R 2 COOH, -SO2R 3 -SOR 3 R 4 -SO2NH2, -SO2NHR 3 -SO2NR 3 R 4 , -NHCOR 3 , -NHSO2R 3 , [ka] It contains an R'' group selected from the group consisting of, R 1 and R 2 However, each is independently hydrogen, halo, or CN. R 3 and R 4 However, each is independent of C 1-6 It is alkyl, A, B, and C are each independently CH or N.

[0203] In a particular embodiment, R'' is -P=O(OH)2, P(=O)R 1 OH, and -(CR 1 R 2 ) is selected from the group consisting of -P=O(OH)2. In a particular embodiment, R'' is -SO2OH, -OSO2OH, -CONHSO2R3 , -SO2R 3 -SOR 3 R 4 -SO2NH2, -SO2NHR 3 -SO2NR 3 R 4 , and -NHSO2R 3 Selected from the group consisting of -OH or -CR. In a particular embodiment, R'' is -OH or -CR 1 R 2 It is OH. In certain embodiments, R'' is -COOH, -CONH2, -CONHR 1 ,-CONR 3 R 4 -CH(COOH)2, -CR 1 R 2 COOH and -NHCOR 3 It is selected from the group consisting of the following.

[0204] Tables 2 and 3 show various exemplary linkers or connecting parts used in the compounds described herein. In some embodiments of formulas (I) to (IIe) or (XI) to (XVIa), the compound comprises one of the linkers or connecting parts shown in Tables 2 and 3. [Table 2] [Table 3-1] [Table 3-2]

[0205] Part of interest (Y) As summarized above, the M6PR or ASGPR-conjugated compounds of this disclosure generally comprise a ligating portion Y of interest. In some embodiments, the portion Y of interest is a chemoselective ligation group or its precursor, and the compounds can find use in the preparation of various conjugates via the conjugation of the chemoselective ligation group to a compatible reactive group of another portion of interest, as described herein, for example.

[0206] Chemoselective ligation groups In certain embodiments of formulas (I) to (XVIa), Y is a chemoselective ligation group or its precursor. A chemoselective ligation group is a group or functional group having a reactive function that enables conjugation of a second part to a compatible group. For example, a chemoselective ligation group (or its precursor) may be one of a pair of groups associated with conjugation chemistry, e.g., azido-alkyne click chemistry, copper-free click chemistry, Staudinger ligation, tetrazine ligation, hydrazine-iso-Pictet-Spengler (HIPS) ligation, cysteine ​​reactive ligation chemistry (e.g., thiol-maleimide, thiol-haloacetamide, or alkyne hydrothiolation), amine-active ester coupling, reductive amination, dialkyl squalate chemistry, etc.

[0207] Chemoselective ligation groups that can be used to link two parts include, but are not limited to, aminos (e.g., N-terminal amino or lysine side chain groups of polypeptides), azides, aryl azides, alkynyls (e.g., ethinyl or cyclooctin or derivatives), active esters (e.g., N-hydroxysuccinimide (NHS) esters, sulfo-NHS esters, or PFP esters or thioesters), haloacetamides (e.g., iodoacetamide or bromoacetamide), chloroacetyl, bromoacetyl, hydrazides, maleimides, vinyl sulfones, 2-sulfonylpyridine, cyano-alkynes, thiols (e.g., cysteine ​​residues), disulfides or protected thiols, isocyanates, isothiocyanates, aldehydes, ketones, alkoxyamines, hydrazides, aminooxy, phosphines, HIPS hydrazinyl-indolyl groups, or aza-HIPS hydrazinyl-pyrrolo-pyridinyl groups, tetrazines, cyclooctene, squalates, and others.

[0208] In some cases, chemoselective ligation groups enable spontaneous conjugation to compatible chemical groups when the two groups are in contact under favorable conditions (e.g., copper-free click chemical conditions). In other cases, chemoselective ligation groups enable conjugation to compatible chemical groups when the two groups are in contact in the presence of a catalyst or other reagent (e.g., copper-catalyzed click chemical conditions).

[0209] 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 and be inserted into the CH, NH, and OH bonds of the second portion.

[0210] In some cases, Y is a reactive functional group or a precursor of a functional group that enables conjugation to a compatible group of the second part. For example, carboxylic acids are precursors of chemoselective ligation groups of active esters.

[0211] In certain embodiments of formulas (Ia) to (XVIa), Y is a reactive moiety capable of forming a covalent bond to the polypeptide (for example, using an amino acid side chain of the polypeptide having a compatible reactive group). The reactive moiety may be referred to as a chemoselective ligation group.

[0212] In certain embodiments of formulas (Ia) to (XVIa), Y is a thioreactive chemoselective ligation group (as shown, for example, in Table 4). In some cases, Y may generate a residue moiety Z resulting from the covalent bonding of a thiol-reactive chemoselective ligation group to one or more cysteine ​​residues of a protein, e.g., Ab.

[0213] In certain embodiments of formulas (Ia) to (XVIa), Y is an amino-reactive chemoselective ligation group (as shown, for example, in Table 4). In some cases, Y may generate a residue moiety Z resulting from the covalent bonding of an amine-reactive chemoselective ligation group to one or more cysteine ​​residues of a protein, e.g., Ab.

[0214] Table 4 shows exemplary chemoselective ligation groups that can be adapted for use in the compounds of this disclosure, and their synthetic precursors. [Table 4-1] [Table 4-2] [Table 4-3]

[0215] Table 4 shows, [ka] This can represent a connecting portion or a point of connection of Y to a connected X portion.

[0216] Exemplary compounds having chemoselective ligation groups This disclosure includes compounds of formulas (Ia) to (Ib) that may include the following: (1) One or more specific M6PR ligands (X) (e.g., ligands X1 to X42 in Table 1, as described herein) or specific ASGPR ligands (X) (e.g., as described herein), (2) A linker including one or more connecting parts (for example, one or more of the connecting parts in Tables 2-3, as described herein), (3) Chemoselective ligation group (Y) (for example, one of the groups in Table 4, as described herein).

[0217] Tables 5–7B show some exemplary M6PR-conjugated compounds of the present disclosure, or their precursors, that contain chemoselective ligation groups. It is understood that the present disclosure includes each of the exemplary compounds in Tables 5–7B with a Y (e.g., as described herein). For example, a conjugate in which the chemoselective ligation group is conjugated to a different Y, such as a biomolecule or small molecule ligand of a target protein.

[0218] Tables 8-9 show some exemplary ASGPR-conjugated compounds of the present disclosure, or their precursors, that contain chemoselective ligation groups. It is understood that the present disclosure includes each of the exemplary compounds in Tables 8-9 with a Y (e.g., as described herein). For example, a conjugate in which the chemoselective ligation group is conjugated to a different Y, such as a biomolecule or small molecule ligand of a target protein.

[0219] The chemoselective ligation group of such a compound can be used to ligate to another Y-molecule of interest (e.g., as described below). It is understood that any of these compounds can be newly prepared to contain an alternative Y-molecule of interest (e.g., as described below) instead of a chemoselective ligation group. In some embodiments, such compounds are referred to as conjugates, e.g., biomolecular conjugates that specifically bind to a target protein. [Table 5-1] [Table 5-2] [Table 5-3] [Table 5-4] [Table 5-5] [Table 5-6] Table 5-7 Table 5-8 Table 5-9 Table 5-10 Table 5-11 Table 6-1 Table 6-2 Table 6-3 Table 6-4 Table 6-5 Table 6-6 Table 6-7 Table 6-8 Table 6-9 Table 6-10 Table 6-11

[0220] In certain embodiments of formulas (Ia) to (Ib), n is 2. In certain embodiments of formulas (Ia) to (Ib), n is 2 and Y is a chemoselective ligation group. In certain embodiments of formulas (Ia) to (Ib), n is 3. In certain embodiments of formulas (Ia) to (Ib), n is 3 and Y is a chemoselective ligation group.

[0221] Exemplary polyvalent M6PR-binding compounds are shown in Tables 7A-7B.

[0222] Examples of polyvalent ASGPR-binding compounds are shown in Tables 8-9. [Table 7-1] [Table 7-2] [Table 7-3] [Table 7-4] [Table 7-5] [Table 7-6] [Table 7-7]

[0223] In certain embodiments of formulas (Ia) to (Ib), n is 2 or greater (e.g., 3 or greater, e.g., 3, 4, 5, or 6 or more), and the linker includes amino acid linking moieties that can be linked together in the sequence to branch and provide linkage to multiple X ligands via their side chains (and optionally terminal groups). In certain embodiments of formula (Ia), n is 3 or greater, and Y is a chemoselective ligation group. In certain embodiments of formula (Ia), n is 4 or greater, and Y is a chemoselective ligation group.

[0224] Table 7B shows exemplary polyvalent compounds containing amino acid residue linkages. [Table 8-1] [Table 8-2] [Table 8-3] [Table 8-4] [Table 8-5] [Table 8-6]

[0225] This disclosure is intended to encompass any one stereoisomer of the compounds described herein. In some cases, the compounds include enantiomers or analogues of the D-mannopyranose ring.

[0226] In a particular embodiment, the compound comprises an L-mannose ring analog, and its structure is: [ka] It has.

[0227] In a particular embodiment, the compound comprises an L-mannose ring and has one of the following structures. [ka]

[0228] Examples of ASGPR-binding compounds of formula (Ib) are shown in Tables 8-9. [Table 9-1] [Table 9-2] [Table 9-3] [Table 9-4] [Table 9-5] [Table 10-1] [Table 10-2]

[0229] Conjugate The compounds of this disclosure may be referred to as conjugates, for example, when the moiety of interest (Y) is a molecule (as described herein). Such conjugates can be prepared by conjugating a chemoselective ligation group of any one of the compounds described herein with a compatible reactive group of molecule Y. The compatible group of child Y can be introduced by modification prior to conjugation, or it may be a group present in the molecule. Alternatively, such a conjugate can be newly prepared, for example, by modifying the starting material of the Y molecule of interest to introduce a linker capable of binding ligand X.

[0230] Aspects of this disclosure include compounds of formula (I) in which the portion of interest Y is selected from small molecules, dyes, fluorophores, monosaccharides, disaccharides, trisaccharides, and biomolecules. In some embodiments, Y is a small molecule that specifically binds to a target molecule, such as a target protein.

[0231] In some embodiments of the compounds of this disclosure, Y is a biomolecule. In some embodiments, the biomolecule is selected from proteins, polynucleotides, polysaccharides, peptides, glycoproteins, lipids, enzymes, antibodies, and antibody fragments. In some embodiments, Y is a biomolecule that specifically binds to a target molecule, such as a target protein.

[0232] The compounds of this disclosure may, in some cases, be referred to as conjugates, for example, when the moiety of interest (Y) is a molecule such as a biomolecule, the conjugate may be derived from a conjugation or coupling reaction between a chemoselective ligation group and a compatible group of the biomolecule. In some embodiments, the biomolecule is conjugated via a naturally occurring group of the biomolecule. In some embodiments, the biomolecule is conjugated via a compatible functional group introduced into the biomolecule prior to chemoselective conjugation. In such cases, the linkage moiety between X and Y incorporates a residual group (e.g., Z) which is a product of the chemoselective ligation chemistry.

[0233] Aspects of the present disclosure include compounds of formula (Ia), wherein the portion of interest Y is a portion that specifically binds to a target molecule, such as a target protein. The target protein may be a membrane-bound protein or an extracellular protein. In some embodiments of the compounds of the present disclosure, Y is a biomolecule that specifically binds to the target protein. The present disclosure provides specific M6PR or ASGPR-binding compounds and their conjugates. In some embodiments, the conjugate comprises the portion of interest Y that specifically binds to the target protein and can be found to be used in methods of cellular uptake or internalization of the target protein via binding to a cell surface receptor and, ultimately, degradation of the target protein.

[0234] In some embodiments, Y is an aptamer that specifically binds to a target molecule, such as a target protein. In some embodiments, Y is a peptide or protein (e.g., a peptide-bonded motif, a protein domain, an engineered polypeptide, or a glycoprotein) that specifically binds to a target molecule, such as a target protein. In some embodiments, Y is an antibody or antibody fragment that specifically binds to a target molecule, such as a target protein. In some embodiments, Y is a polynucleotide or oligonucleotide that specifically binds to a target molecule, such as a target protein or a target nucleic acid.

[0235] 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., M6PR or ASGPR). In some embodiments, one Y biomolecule is conjugated to a single moiety (X) n -L)- is conjugated, and when n=1, (X n -L)- groups are called monovalent, and when n>1, (X n The -L)- group is referred to as polyvalent (e.g., divalent, trivalent, etc.). In some embodiments of formula (Ia) where Y is a biomolecule, Y has two or more (X n -L)- may be conjugated, each (X nIt is understood that the -L)- group can be monovalent or polyvalent (e.g., divalent, trivalent, etc.). In such cases, the linked (X) group to the biomolecule n The ratio of -L)- groups can be said to be 2 or more.

[0236] Therefore, the following equation (IVa) is conjugated: [ka] Or a pharmaceutically acceptable salt thereof is provided herein. During the ceremony, X is a portion that binds to cell surface M6PR (e.g., as described herein) or to cell surface ASGPR (e.g., as described herein), L is a linker (for example, as described herein), n is an integer between 1 and 500 (for example, between 1 and 5). m is an integer between 1 and 80. Z is a residue portion resulting from the covalent bonding of a chemoselective ligation group (Y) to P. P is a biomolecule (for example, a biomolecule that specifically binds to a target protein as described herein).

[0237] In some embodiments of formula (IVa), L is a linker of formulas (IIa) to (IId) (e.g., as described herein). In some embodiments of formula (IVa), Xn-LZ is derived from a compound of formulas (XI) to (XVIa) (e.g., as described herein), and Y is a chemoselective ligation group.

[0238] In formula (IVa), Z can be any convenient residue portion resulting from the covalent bonding or conjugation of a chemoselective ligation group (Y) to a compatible reactive group of a biomolecule (P). In some cases, the compatible reactive group of a biomolecule (P) is a group that can naturally become part of the biomolecule. In some cases, the compatible reactive group of a biomolecule (P) is introduced or incorporated into the biomolecule before conjugation. In such cases, the biomolecule (P) can be a modified version of the biomolecule. For example, a functional group of a biomolecule (e.g., an amino group, a carboxylic acid group, or a thiol group) can be modified to introduce a compatible chemoselective ligation group (e.g., using a chemical reagent such as a 2-haloacetyl reagent, or 2-iminothiolane, or via coupling of a linker group containing a chemoselective ligation group such as an azide or alkyne).

[0239] In some embodiments of formula (IVa), L is the linker of formula (IIa) (for example, as described herein). In a particular embodiment of formula (IVa), Z is [ka] Selected from the group consisting of, [ka] However, this represents the connection point to linker L, [ka] This represents the connection point to P, W is CH2, N, O, or S. P is a polypeptide.

[0240] In a particular embodiment of formula (IVa), Z is [ka] Selected from the group consisting of, [ka] However, this represents the connection point to L, [ka] This represents the connection point to P, P is a polypeptide.

[0241] In a particular embodiment of formula (IVa), Z is [ka] Selected from the group consisting of, [ka] However, this represents the connection point to L, [ka] This represents a binding site to P. In some embodiments, P is a polypeptide.

[0242] In a particular embodiment, n is 1. In a particular embodiment, n is 2. In a particular embodiment, n is 3. In a particular embodiment, n is 4. In a particular embodiment, n is 5.

[0243] In yet another embodiment, a conjugate of formula (IVa) is provided herein, where L is a linker of the following formula (IIe): [ka] In the formula, L 1 , L 2 , L 3 , L 4 , L 5 , and n are defined herein.

[0244] In a particular embodiment, L is selected from the linkers in Tables 1 and 2.

[0245] In another embodiment, the following conjugate of formula (IVb) is provided herein: [ka] During the ceremony, X is a portion that binds to cell surface M6PR (e.g., as described herein) or to cell surface ASGPR (e.g., as described herein), m is an integer between 1 and 80. Z [ka] It is a part that has the structure, [ka] This represents the connection point to X, [ka] represents a binding site to P, where P is a biomolecule (for example, a polypeptide as described herein).

[0246] In certain embodiments of the conjugate of formula (IVa) or (IVb), P is a peptide or protein as defined herein.

[0247] In certain embodiments of the conjugate of formula (IVa) or (IVb), P is selected from an antibody, an antibody fragment (e.g., an antigen-binding fragment of an antibody), a chimeric fusion protein, an engineered protein domain, a D protein binder for a target protein, and a peptide.

[0248] In certain embodiments of the conjugate of formula (IVa) or (IVb), P is an antibody or antibody fragment (Ab) as defined herein.

[0249] Therefore, in another embodiment, the following equation (Va) is conjugated: [ka] Or a pharmaceutically acceptable salt thereof is provided herein. During the ceremony, X is a portion that binds to cell surface M6PR (e.g., as described herein) or to cell surface ASGPR (e.g., as described herein), L is a linker (for example, as described herein), n is an integer between 1 and 5. m is an integer between 1 and 80. Z [ka] Ab is a residue portion resulting from the covalent bonding of a chemoselective ligation group (Y) to a compatible group, where Ab is an antibody or antibody fragment.

[0250] In some embodiments of formula (Va), L is the linker of formula (IIa) (for example, as described herein).

[0251] In a particular embodiment of equation (Va), Z is [ka] Selected from the group consisting of, [ka] However, this represents the connection point to L, [ka] but, [ka] Represents the connection point to, W is CH2, N, O, or S. [ka] However, it is an antibody.

[0252] In a particular embodiment, Z is [ka] Selected from the group consisting of, [ka] However, this represents the connection point to L, [ka] but, [ka] Represents the connection point to, [ka] However, it is an antibody.

[0253] In a particular embodiment, Z is [ka] Selected from the group consisting of, [ka] However, this represents the connection point to L, [ka] but, [ka] Represents the connection point to, [ka] However, it is an antibody.

[0254] In another embodiment, the following equation (Va) is conjugated: [ka] Or a pharmaceutically acceptable salt thereof is provided herein. X is a portion that binds to cell surface M6PR (e.g., as described herein) or to cell surface ASGPR (e.g., as described herein), L is the linker in equation (IIa) below: [ka] , and in the formula, Each L 1 However, they became independent, [ka] , -C 1-6 -Alkilen-, -(OCH2CH2) k -, or -(OCH2CH2) k -(CH2) v -and, Each L 3 However, they became independent, [ka] , or -(OCH2CH2) q -and, Each L 4 However, independently, -OCH2CH2-, [ka] And, Each L 5 However, it became independent, -NHCO-C 1-6 -Alkylene- or -(OCH2CH2) r -and, Each L 6 However, it became independent, -NHCO-C 1-6 -Alkylene- or -(OCH2CH2) s -and, Each L 7 However, it became independent, -NHCO-C 1-6 -Alkilen-, -(OCH2CH2) t-, or -OCH2- p, q, r, s, and t are each independently integers between 1 and 20, a is 1 or 2, b, c, d, e, f, and g are each independently 0, 1, or 2, and u, v, w, x, y, and z are each independently integers between 1 and 10. If n is an integer from 1 to 5 and d is 0, if n is 1 and d is 1, if n is an integer from 1 to 3 and d is 2, if n is an integer from 1 to 5, m is an integer between 1 and 80. Z [ka] Selected from the group consisting of, [ka] However, this represents the connection point to L, [ka] but, [ka] Represents the connection point to, [ka] However, it is an antibody.

[0255] In another embodiment, the following equation (Va) is conjugated: [ka] Or a pharmaceutically acceptable salt thereof is provided herein, in the formula, X is a portion that binds to cell surface M6PR (e.g., as described herein) or to cell surface ASGPR (e.g., as described herein), L is the linker in equation (IIb) below: [ka] During the ceremony, L 1 but, [ka] And, L 2 However, -(OCH2CH2) p -and, L 3 However, -NHCO-C 1-6 -Alkilen-, p is an integer between 1 and 20, a is 1, and b and c are independently either 0 or 1. n is 2, m is an integer between 1 and 80. Z [ka] Selected from the group consisting of, [ka] However, this represents the connection point to L, [ka] but, [ka] Represents the connection point to, [ka] However, it is an antibody.

[0256] In another embodiment, the following equation (Va) is conjugated: [ka] Or a pharmaceutically acceptable salt thereof is provided herein, in the formula, X is a portion that binds to cell surface M6PR (e.g., as described herein) or to cell surface ASGPR (e.g., as described herein), L is the linker in equation (IIb) below: [ka] During the ceremony, L 1 but, [ka] And, L 2 However, -(OCH2CH2) p -and, L 3 However, -NHCO-C 1-6 -Alkilen-, p is an integer between 1 and 20, a is 1, and b and c are independently either 0 or 1. n is 2, m is an integer between 1 and 80. Z [ka] Selected from the group consisting of, [ka] However, this represents the connection point to L, [ka] but, [ka] Represents the connection point to, [ka] However, it is an antibody.

[0257] In another embodiment, the following equation (Va) is conjugated: [ka] Or a pharmaceutically acceptable salt thereof is provided herein, in the formula, X is a portion that binds to cell surface M6PR (e.g., as described herein) or to cell surface ASGPR (e.g., as described herein), L is the linker in equation (IIc) below: [ka] During the ceremony, L 1 but, [ka] And, L 2 but, [ka] And, L 3 However, -NHCO-C 1-6 -Alkylene- or -(OCH2CH2) p -and, L 4 However, -NHCO-C 1-6 -Alkylene- or -(OCH2CH2) q -and, p and q are each independent integers between 1 and 20, a is 1, b, c, and d are each independent 0 or 1, and w and u are each independent integers between 1 and 10. [ka] This represents the connection point to X, [ka] However, L 2 Represents the connection point to, n is 2, m is an integer between 1 and 80. Z [ka] Selected from the group consisting of, [Chemical] represents the binding point to L, [Chemical] is [Chemical] represents the binding point to [Chemical] is an antibody.

[0258] In another aspect, a conjugate of the following formula (Va): [Chemical] or a pharmaceutically acceptable salt thereof is provided herein, wherein X is a moiety that binds to cell surface M6PR (e.g., as described herein) or a moiety that binds to cell surface ASGPR (e.g., as described herein), L is a linker of the following formula (IIc): [Chemical] wherein L 1 is [Chemical] is L 2 is [Chemical] is L 3 is -NHCO-C 1-6 -alkylene- or -(OCH2CH2) p - is L 4 is -NHCO-C 1-6 -alkylene- or -(OCH2CH2)q -and, p and q are each independent integers between 1 and 20, a is 1, b, c, and d are each independent 0 or 1, and w and u are each independent integers between 1 and 10. [ka] This represents the connection point to X, [ka] However, L 2 Represents the connection point to, n is 2, m is an integer between 1 and 80. Z [ka] Selected from the group consisting of, [ka] However, this represents the connection point to L, [ka] but, [ka] Represents the connection point to, [ka] However, it is an antibody.

[0259] In another embodiment, the following equation (Va) is conjugated: [ka] Or a pharmaceutically acceptable salt thereof is provided herein, in the formula, X is a portion that binds to cell surface M6PR (e.g., as described herein) or to cell surface ASGPR (e.g., as described herein), L is a linker of the following formula (IId):

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

Chem.

[0260] In another embodiment, the following expression (Vb) is conjugated: [ka] Or a pharmaceutically acceptable salt thereof is provided herein, in the formula, X is a portion that binds to cell surface M6PR (e.g., as described herein) or to cell surface ASGPR (e.g., as described herein), m is an integer between 1 and 80. Z [ka] It is a part that has the structure, [ka] This represents the connection point to X, [ka] but, [ka] Represents the connection point to, [ka] However, it is an antibody.

[0261] In certain embodiments of the conjugates of formulas (IVa), (IVb), (Va), and / or (Vb), X is an M6PR binding moiety as described herein, for example, X of formulas (XI) to (XVIa), or formulas (IIIa), (IIIb), (IIIc), or (IIId). In certain embodiments of the conjugates of formulas (IVa), (IVb), (Va), and / or (Vb), X is an M6PR binding moiety as described in Table 1 and any one of the ligands and compounds in Tables 5 to 7.

[0262] In certain embodiments of the conjugates of formulas (IVa), (IVb), (Va), and / or (Vb), X is an ASGPR binding moiety, for example, as described herein, and is X of formulas (IIIa'), (IIIa''), (IIIb'), (IIIb''), (IIIc'), (IIIc''), (IIId'), or (IIId''). In certain embodiments of the conjugates of formulas (IVa), (IVb), (Va), and / or (Vb), X is an ASGPR binding moiety, as described in any one of the compounds in Tables 8-9. In certain embodiments of the conjugates of formulas (IVa), (IVb), (Va), and / or (Vb), X is X of formulas (IIIa'), (IIIa''), (IIIb'), or (IIIb''). In certain embodiments of the conjugates of formulas (IVa), (IVb), (Va), and / or (Vb), X is X of formula (IIIc'), (IIIc''), (IIId'), or (IIId''). In certain embodiments of the conjugates of formulas (IVa), (IVb), (Va), and / or (Vb), X is X of formula (IIIa') or (IIIa''). In certain embodiments of the conjugates of formulas (IVa), (IVb), (Va), and / or (Vb), X is X of formula (IIIb') or (IIIb''). In certain embodiments of the conjugates of formulas (IVa), (IVb), (Va), and / or (Vb), X is X of formula (IIIc') or (IIIc''). In certain embodiments of the conjugates of formulas (IVa), (IVb), (Va), and / or (Vb), X is X of formula (IIId') or (IIId'').

[0263] In a particular embodiment, the conjugate of formulas (IVa), (IVb), (Va), and / or (Vb) is: [ka] [ka] Selected from the group consisting of, or pharmaceutically acceptable salts thereof, During the ceremony, m is an integer between 1 and 80. [ka] However, it is an antibody.

[0264] In a particular embodiment, the conjugate of formulas (IVa), (IVb), (Va), and / or (Vb) is: [ka] Selected from the group consisting of, or pharmaceutically acceptable salts thereof, During the ceremony, m is an integer between 1 and 80. [ka] However, it is an antibody.

[0265] In a particular embodiment, the conjugate of formulas (IVa), (IVb), (Va), and / or (Vb) is: [ka] Selected from the group consisting of or pharmaceutically acceptable salts thereof, During the ceremony, m is an integer between 1 and 80. [ka] However, it is an antibody.

[0266] In a particular embodiment, formulas (IVa), (IVb), (Va), and / or The conjugate of (Vb) has the following equation (IX), [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, m is an integer between 1 and 4. [ka] However, it is an antibody.

[0267] In a particular embodiment, the conjugate of formulas (IVa), (IVb), (Va), and / or (Vb) has the following formula (X): [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, m is an integer between 1 and 4. [ka] However, it is an antibody.

[0268] In a particular embodiment, the conjugate of formulas (IVa), (IVb), (Va), and / or (Vb) has the following formula (XI), [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, m is an integer between 1 and 4. [ka] However, it is an antibody.

[0269] In a particular embodiment, the conjugate of formulas (IVa), (IVb), (Va), and / or (Vb) has the following formula (XII), [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, m is an integer between 1 and 4. [ka] However, it is an antibody.

[0270] In another embodiment, the following equation (VIa) is conjugated: [ka] Or a pharmaceutically acceptable salt thereof is provided herein. During the ceremony, o is an integer between 1 and 10. m is an integer between 1 and 80. L is the linker, Z [ka] Selected from the group consisting of, [ka] However, this represents the connection point to L, [ka] This represents the connection point to P, P is a polypeptide, X [ka] And, R L However, it is --O-, -NH-, -S-, or -CH2-, R'' is -OH, -CR 1 R 2 OH, -P=O(OH)2, P(=O)R 1 OH, -PH(=O)OH, -(CR 1 R 2 )-P=O(OH)2, -SO2OH, -S(O)OH, -OSO2OH, -COOH, -CONH2, -CONHR 3 ,-CONR 3 R 4 -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 -SO2NH2, -SO2NHR 3 -SO2NR 3 R 4 , -SO2NHCOR 3 , -NHCOR 3 ,-NHC(O)NHS(O)2R 3 , -NHSO2R 3 , [ka] Selected from the group consisting of, j is an integer between 1 and 3. R 1 and R 2 However, each is independently hydrogen, halo, or CN. R 3 and R 4 However, each is independent of C 1-6 It is alkyl, A, B, and C are each independently CH or N. D is either O or S independently, and n, L, and Y are as described for equation (Ia), except R L However, --O-, and R'' is, [ka] And if B and C are N, then j is 2, however R L is -O-, and R'' is -CR 1 R 2 If it is COOH, then R 1 and R 2 However, neither of them is hydrogen.

[0271] In another embodiment, the following equation (VIa) is conjugated: [ka] Or a pharmaceutically acceptable salt thereof is provided herein. During the ceremony, o is an integer between 1 and 10. m is an integer between 1 and 80. L is the linker, Z [ka] Selected from the group consisting of, [ka] However, this represents the connection point to L, [ka] This represents the connection point to P, P is a polypeptide, X [ka] And, R L However, it is --O-, -NH-, -S-, or -CH2-, R'' is -P=O(OH)2, P(=O)R 1 OH, -PH(=O)OH, -(CR 1 R 2 )-P=O(OH)2, -S(O)OH, -OSO2OH, -CONH2, -CONHR 3 ,-CONR 3 R 4 -CONH(OH), -CONH(OR 3 ), -CONHSO2R 3 -CONHSO2NR 3 R 4 ,-CR 1 R 2 COOH, -SO2R 3 -SOR 3 R 4 -SO2NH2, -SO2NHR 3 -SO2NR 3 R 4 , -SO2NHCOR3 , -NHCOR 3 , -NHC(O)NHS(O)2R 3 , -NHSO2R 3 , [Chemical formula] selected from the group consisting of, j is an integer from 1 to 3, R 1 and R 2 are each independently hydrogen, halo, or CN, R 3 and R 4 are each independently C 1-6 alkyl, A, B, and C are each independently CH or N, D is each independently O or S, and n, L, and Y are as described for formula (Ia), provided that R L is -O-, and R’’ is [Chemical formula] and when B and C are N, j is 2, provided that R L is -O-, and R’’ is -CR 1 R 2 COOH, then R 1 and R 2 are not both hydrogen.

[0272] In another aspect, the conjugate of the following formula (VIa): [Chemical formula] or a pharmaceutically acceptable salt thereof is provided herein, wherein, o is an integer from 1 to 10, m is an integer from 1 to 80, L is a linker, Z is [Chemical formula] Selected from the group consisting of, [ka] However, this represents the connection point to L, [ka] This represents the connection point to P, P is a polypeptide, X [ka] or [ka] , and R L However, it is --O-, -NH-, -S-, or -CH2-, R'' is -P=O(OH)2, P(=O)R 1 OH, -PH(=O)OH, -(CR 1 R 2 )-P=O(OH)2, -S(O)OH, -OSO2OH, -CONH(OH), -CONH(OR 3 )-CONHSO2R 3 -CONHSO2NR 3 R 4 , -SO2R 3 -SOR 3 R 4 -SO2NH2, -SO2NHR 3 -SO2NR 3 R 4 , -SO2NHCOR 3 , -NHCOR 3 ,-NHC(O)NHS(O)2R 3 , -NHSO2R 3 , [ka] Selected from the group consisting of, j is an integer between 1 and 3. R 1 and R 2However, each is independently hydrogen, halo, or CN. R 3 and R 4 However, each is independent of C 1-6 It is alkyl, A, B, and C are each independently CH or N. Each D is independently either O or S.

[0273] In certain embodiments, conjugates having the linker structures described herein have a weaker binding affinity to cell surface receptors. While not bound by any particular mechanism or theory, such weaker binding affinity may be modified to a longer half-life of the conjugate, which may be useful for regulating (e.g., modifying) the pharmacokinetic properties of the conjugates described herein. In certain embodiments, such weakly binding conjugates still have sufficiently robust uptake.

[0274] The term "pharmaceutically acceptable" means that it is approved by a federal or state regulatory authority for use in animals, and more specifically in humans, or is listed in the United States Pharmacopeia, the European Pharmacopeia, or any other generally accepted pharmacopoeia.

[0275] The term "pharmaceutically acceptable salt" refers to a salt that is suitable for use in contact with human and lower animal tissues without excessive toxicity, irritation, or allergic reactions. pharmaceutically acceptable salts are well known in the art. For example, SMBerge 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 separately by reacting the free basic functional group 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 such as hydrochloric acid, hydrobromic acid, or phosphoric acid, or organic acids such as acetic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid. Other pharmaceutically acceptable salts include, but are not limited to, adipine, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, citrate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, gluconate, 2-hydroxyethanesulfonate, lactate, laurate, malate, maleate, malonate, methanesulfonate, oleate, oxalate, palmitate, phosphate, propionate, stearate, succinate, sulfate, tartrate, p-toluenesulfonate, and valerate. Typical alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, or magnesium salts. Further pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium, and counterions, such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, and alkyl groups having 1 to 6 carbon atoms (e.g., C 1-6 It includes amine cations formed using alkyl, sulfonate, and aryl sulfonate compounds.

[0276] Polypeptide (P), for example, an antibody (Ab) and a compound (Xn-LY), conjugates can be prepared using various 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. This disclosure further intends that the conjugates described herein can be prepared using any suitable method disclosed in the art (see, for example, Bioconjugate Techniques (Hermanson ed., 2d ed. 2008)).

[0277] In certain embodiments of the conjugate described herein, L is bound to a lysine residue of P via an amide bond. In certain embodiments of the conjugate described herein, L is bound to a cysteine ​​residue of P via a thioether bond. In certain embodiments of the conjugate described herein, L is bound to a lysine residue of Ab via an amide bond, as described above. In certain embodiments of the conjugate described herein, L is bound to a cysteine ​​residue of Ab via a thioether bond, as described above. In certain embodiments of the conjugate described herein, L is bound to two cysteine ​​residues of Ab via two thioether bonds, the two cysteine ​​residues derived from an open cysteine-cysteine ​​disulfide bond of Ab, as described above. In certain embodiments, the open cysteine-cysteine ​​disulfide bond is an interchain disulfide bond.

[0278] In certain embodiments of the conjugate described herein, when L is bound to a lysine residue of P via an amide bond, m is an integer from 1 to 80. In certain embodiments of the conjugate described herein, when L is bound to a cysteine ​​residue of P via a thioether bond, m is an integer from 1 to 8.

[0279] In certain embodiments, conjugation to polypeptide P or antibody Ab may be via site-specific conjugation. Site-specific conjugation may result in, for example, a uniform loading and minimization of a conjugate subpopulation with potentially altered antigen binding or pharmacokinetics. In certain embodiments, for example, the conjugation may involve operations such as cysteine ​​substitution at a position on the polypeptide or antibody, e.g., providing a reactive thiol group, which does not disrupt the folding and assembly of the polypeptide or antibody, or alters the polypeptide or antigen binding (e.g., Junutula et al., J.Immunol.Meth.2008;332:41-52, and Junutula et al., Nature). See Biotechnol.2008;26:925-32 and also see WO2006 / 034488 (which is incorporated herein by reference in its entirety). Another non-limiting approach involves co-translationally inserting selenocysteine ​​into a polypeptide or antibody sequence by recoding the stop codon UGA from termination to selenocysteine ​​insertion, enabling site-specific covalent conjugation of selenocysteine ​​at the nucleophilic selenool group in the presence of other native amino acids (e.g., Hofer). See et al., Proc. Natl. Acad. Sci. USA 2008;105:12451-56, and Hofer et al., Biochemistry 2009;48(50):12047-57. Further non-limiting techniques that enable site-specific conjugation to polypeptides or antibodies include, for example, the manipulation of non-natural amino acids including p-acetylphenylalanine (p-acetyl-Phe), p-azidomethyl-N-phenylalanine (p-azidomethyl-Phe), and azidridine (azido-Lys) at specific linkage sites, and may further include manipulating proprietary functional tags such as LPXTG, LLQGA, sialic acid, and GlcNac for enzyme-mediated conjugation. See Jackson, Org. Process Res. Dev. 2016;20:852-866, and Tsuchikama and An, Protein Cell 2018;9(1):33-46, the contents of each of these are incorporated together by reference. See also US2019 / 0060481A1 and US2016 / 0060354A1, the contents of each of these are incorporated together by reference. All such methodologies are intended to be used in connection with the preparation of the conjugates described herein.

[0280] The loading of compounds of formulas (Ia) and (Ib) onto polypeptides (e.g., antibodies) as described herein is represented by "m" in formulas (IVa), (IVb), (Va), and / or (Vb), 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 the conjugate. The number of "X" parts (e.g., M6P parts) per unit of "Xn-L-" or "Xn-" is represented by "n" in formulas (IVa), (IVb), (Va), and / or (Vb). As used herein, the term "valence(s)" refers to the number of "X" parts ("n") per unit. It should be understood that loading, or DAR, is not necessarily equivalent to the number of "X" parts per conjugate molecule. For example, if there is one "X" part per unit (n=1, valence "1") and one "Xn-L-" unit per conjugate (m=1), then there are 1 × 1 = 1 "X" part per conjugate. However, if there are two "X" parts per unit (n=2, valence "2") and four "Xn-L-" units per conjugate (m=4), then there are 2 × 4 = 8 "X" parts per conjugate. Therefore, for the conjugates described herein, the total number of "X" parts per conjugate molecule is n × m. As used herein, the term "total valence(s)" refers to the total number of "X" parts per conjugate molecule (n x m; total valence).

[0281] The DAR (load) is in the range of 1 to 80 units per conjugate. The conjugates provided herein may include aggregates of conjugated polypeptides, antibodies, or antigen-binding fragments with a unit range, e.g., 1 to 80. The average number of units per polypeptide or antibody in the preparation of conjugates from a conjugation reaction can be characterized by conventional means such as mass spectrometry. The quantitative distribution of DAR (load) with respect to m can also be determined. In some cases, the separation, purification, and characterization of homogeneous conjugates where m is a specific value can be achieved by means such as electrophoresis.

[0282] In certain embodiments, the DAR of the conjugate provided herein is in the range of 1 to 80. In certain embodiments, the DAR of the conjugate provided herein is in the range of 1 to 70. In certain embodiments, the DAR of the conjugate provided herein is in the range of 1 to 60. In certain embodiments, the DAR of the conjugate provided herein is in the range of 1 to 50. In certain embodiments, the DAR of the conjugate provided herein is in the range of 1 to 40. In certain embodiments, the DAR of the conjugate provided herein is in the range of 1 to 35. In certain embodiments, the DAR of the conjugate provided herein is in the range of 1 to 30. In certain embodiments, the DAR of the conjugate provided herein is in the range of 1 to 25. In certain embodiments, the DAR of the conjugate provided herein is in the range of 1 to 20. In certain embodiments, the DAR of the conjugate provided herein is in the range of 1 to 18. In certain embodiments, the DAR of the conjugate provided herein is in the range of 1 to 15. In certain embodiments, the DAR of the conjugate provided herein is in the range of 1 to 12. In certain embodiments, the DAR of the conjugate provided herein is in the range of 1 to 10. In certain embodiments, the DAR of the conjugate provided herein is in the range of 1 to 9. In certain embodiments, the DAR of the conjugate provided herein is in the range of 1 to 8. In certain embodiments, the DAR of the conjugate provided herein is in the range of 1 to 7. In certain embodiments, the DAR of the conjugate provided herein is in the range of 1 to 6. In certain embodiments, the DAR of the conjugate provided herein is in the range of 1 to 5. In certain embodiments, the DAR of the conjugate provided herein is in the range of 1 to 4. In certain embodiments, the DAR of the conjugate provided herein is in the range of 1 to 3. In certain embodiments, the DAR of the conjugate provided herein is in the range of 2 to 12.In certain embodiments, the DAR of the conjugate provided herein is in the range of 2 to 10. In certain embodiments, the DAR of the conjugate provided herein is in the range of 2 to 9. In certain embodiments, the DAR of the conjugate provided herein is in the range of 2 to 8. In certain embodiments, the DAR of the conjugate provided herein is in the range of 2 to 7. In certain embodiments, the DAR of the conjugate provided herein is in the range of 2 to 6. In certain embodiments, the DAR of the conjugate provided herein is in the range of 2 to 5. In certain embodiments, the DAR of the conjugate provided herein is in the range of 2 to 4. In certain embodiments, the DAR of the conjugate provided herein is in the range of 3 to 12. In certain embodiments, the DAR of the conjugate provided herein is in the range of 3 to 10. In certain embodiments, the DAR of the conjugate provided herein is in the range of 3 to 9. In certain embodiments, the DAR of the conjugate provided herein is in the range of 3 to 8. In a particular embodiment, the DAR of the conjugate provided herein is in the range of 3 to 7. In a particular embodiment, the DAR of the conjugate provided herein is in the range of 3 to 6. In a particular embodiment, the DAR of the conjugate provided herein is in the range of 3 to 5. In a particular embodiment, the DAR of the conjugate provided herein is in the range of 3 to 4.

[0283] In a particular embodiment, the DAR of the conjugate provided herein is in the range of 1 to about 8, about 2 to about 6, about 3 to about 5, about 3 to about 4, about 3.1 to about 3.9, about 3.2 to about 3.8, about 3.2 to about 3.7, about 3.2 to about 3.6, about 3.3 to about 3.8, or about 3.3 to about 3.7.

[0284] In certain embodiments, the DAR of the conjugate provided herein is about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12 or more. In some embodiments, the DAR of the conjugate 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.

[0285] In some embodiments, the DAR of the conjugates provided herein is in the range of 2-20, 2-19, 2-18, 2-17, 2-16, 2-15, 2-14, or 2-13. In some embodiments, the DAR of the conjugates provided herein is in the range of 3-20, 3-19, 3-18, 3-17, 3-16, 3-15, 3-14, or 3-13. In some embodiments, the DAR of the conjugates provided herein is about 1. In some embodiments, the DAR of the conjugates provided herein is about 2. In some embodiments, the DAR of the conjugates provided herein is about 3. In some embodiments, the DAR of the conjugates provided herein is about 4. In some embodiments, the DAR of the conjugates provided herein is about 3.8. In some embodiments, the DAR of the conjugates provided herein is about 5. In some embodiments, the DAR of the conjugates provided herein is about 6. In some embodiments, the DAR of the conjugate provided herein is about 7. In some embodiments, the DAR of the conjugate provided herein is about 8. In some embodiments, the DAR of the conjugate provided herein is about 9. In some embodiments, the DAR of the conjugate provided herein is about 10. In some embodiments, the DAR of the conjugate provided herein is about 11. In some embodiments, the DAR of the conjugate provided herein is about 12. In some embodiments, the DAR of the conjugate provided herein is about 13. In some embodiments, the DAR of the conjugate provided herein is about 14. In some embodiments, the DAR of the conjugate provided herein is about 15. In some embodiments, the DAR of the conjugate provided herein is about 16. In some embodiments, the DAR of the conjugate provided herein is about 17. In some embodiments, the DAR of the conjugate provided herein is about 18.In some embodiments, the DAR of the conjugate provided herein is about 19. In some embodiments, the DAR of the conjugate provided herein is about 20.

[0286] In some embodiments, the DAR of the conjugate provided herein is about 25. In some embodiments, the DAR of the conjugate provided herein is about 30. In some embodiments, the DAR of the conjugate provided herein is about 35. In some embodiments, the DAR of the conjugate provided herein is about 40. In some embodiments, the DAR of the conjugate provided herein is about 50. In some embodiments, the DAR of the conjugate provided herein is about 60. In some embodiments, the DAR of the conjugate provided herein is about 70. In some embodiments, the DAR of the conjugate provided herein is about 80.

[0287] In certain embodiments, fewer units than the theoretical maximum are conjugated to a polypeptide, such as an antibody, during the conjugation reaction. The polypeptide may contain, for example, lysine residues that do not react with the compound or linker reagent. Generally, for example, antibodies do not contain many free and reactive cysteinethiol groups that can be linked to drug units; in fact, most cysteinethiol residues in antibodies exist as disulfide crosslinks. In certain embodiments, the antibody may be reduced with a reducing agent such as dithiothreitol (DTT) or tricarbonylethylphosphine (TCEP) under partially or entirely reducing conditions to generate reactive cysteinethiol groups. In certain embodiments, the antibody is subjected to denaturing conditions to reveal reactive nucleophiles such as lysine or cysteine. In some embodiments, the compound is conjugated via lysine residues on the antibody. In some embodiments, the linker unit or drug unit is conjugated via cysteine ​​residues on the antibody.

[0288] In certain embodiments, the amino acid that binds to the unit is located in the heavy chain of the antibody. In certain embodiments, the amino acid that binds to the unit is located in the light chain of the antibody. In certain embodiments, the amino acid that binds to the unit is located in the hinge region of the antibody. In certain embodiments, the amino acid that binds to the unit is located in the Fc region of the antibody. In certain embodiments, the amino acid that binds to the unit is located in the constant region of the antibody (e.g., CH1, CH2, or CH3 of the heavy chain, or CH1 of the light chain). In yet another embodiment, the amino acid that binds to the unit or drug unit is located in the VH framework region of the antibody. In yet another embodiment, the amino acid that binds to the unit is located in the VL framework region of the antibody.

[0289] The DAR (load) of the conjugate can be controlled in different ways, for example, by (i) limiting the molar excess of the compound or conjugation reagent compared to the polypeptide, (ii) limiting the conjugation reaction time or temperature, (iii) partial or limited reduction conditions for cysteine ​​thiol modification, or (iv) manipulating the amino acid sequence of the polypeptide by recombinant technology such as modifying the number and position of cysteine ​​residues to adjust the number and / or position of linker-drug binding (e.g., thiomab prepared as disclosed in WO2006 / 034488 (which is incorporated herein by reference in its entirety)).

[0290] It should be understood that the preparation of conjugates described herein may result in a mixture of conjugates having a distribution of one or more units bound to polypeptides, such as antibodies. Individual conjugate molecules may be identified in the mixture by mass spectrometry and separated by HPLC, for example, hydrophobic interaction chromatography, including such methods known in the art. In certain embodiments, a homogeneous conjugate having a single DAR (load) value may be isolated from the conjugation mixture by electrophoresis or chromatography.

[0291] Polypeptide (P): In certain embodiments, the conjugate polypeptide (P) comprises a polypeptide that binds to a soluble (e.g., secreted) polypeptide of interest. In certain embodiments, for example, the polypeptide of interest is a ligand that binds to a cell surface receptor, and P comprises a ligand-binding portion of the cell surface receptor, for example, the extracellular domain of the cell surface receptor, for example, the ligand-binding domain of the extracellular domain of the cell surface receptor. In certain embodiments, the polypeptide of interest is a cell surface receptor, and P comprises a ligand that binds to the cell surface receptor, or the receptor-binding portion of the ligand.

[0292] It will be understood by those skilled in the art that a polypeptide (P) that binds to a polypeptide of interest is conjugated as “conjugated” in this context. For example, P, e.g., an antibody, or a conjugate described herein containing such P may bind to other polypeptides with a lower affinity, generally determined by immunoassay or other assays known in the art. In certain embodiments, P, or a conjugate described herein containing such P that specifically binds to a polypeptide of interest, binds to a polypeptide of interest having an affinity of at least 2-to-5, 2.5-to-5, 3-to-5, 4-to-5 or more than the affinity when P or the conjugate binds to another polypeptide. In another particular embodiment, P, or a conjugate described herein containing such P, does not specifically bind to polypeptides other than the polypeptide of interest. In certain embodiments, P, or a conjugate described herein containing P, binds with an affinity (K) of 20 mM or less. d ) specifically binds to the polypeptide of interest. In certain embodiments, such binding has affinity (K) of about 20 mM, about 10 mM, about 1 mM, about 100 μM, about 10 μM, about 1 μM, about 100 nM, about 10 nM, or less than about 1 nM. d) has. Unless otherwise specified, "binds," "binds to," "specifically binds," or "specifically binds to" are interchangeable in this context.

[0293] In one particular embodiment, for example, the polypeptide of interest is a cell surface receptor, and P comprises a cell surface protein, such as an antibody that binds to the extracellular domain of the cell surface receptor. In another embodiment, for example, the polypeptide of interest is a soluble (e.g., secreted) polypeptide of interest, such as a ligand for a cell surface receptor polypeptide, and P comprises an antibody that binds to the ligand.

[0294] Polypeptides may contain L-amino acids, D-amino acids, or both, and may contain any of the various amino acid modifications or analogs known in the art. Useful modifications include, for example, terminal acetylation, amidation, and methylation.

[0295] In certain embodiments, the polypeptide (P) contains about 10, about 20, about 30, about 40, about 50, about 100, about 150, about 200, about 250, about 300, about 350, about 400, about 450, about 500, about 550, about 600, about 650, about 700, about 750, about 800, about 850, about 900, or about 950 amino acids.

[0296] In a particular embodiment, the polypeptide (P) contains approximately 10–50, approximately 50–100, approximately 100–150, approximately 150–200, approximately 200–250, approximately 250–300, approximately 300–350, approximately 350–400, approximately 400–450, approximately 450–500, approximately 500–600, approximately 600–700, approximately 700–800, approximately 800–900, or approximately 900–1000 amino acids.

[0297] In certain embodiments, the conjugate comprises an antibody Ab. In certain embodiments, Ab is a monoclonal antibody. In certain embodiments, Ab is a human antibody. In certain embodiments, Ab is a humanized antibody. In certain embodiments, Ab is a chimeric antibody. In certain embodiments, Ab is a full-length antibody comprising two heavy chains and two light chains. In certain embodiments, Ab is an IgG antibody, such as an IgG1, IgG2, IgG3, or IgG4 antibody. In certain embodiments, Ab is a single-chain antibody. In yet another embodiment, Ab is an antigen-binding fragment of an antibody, such as a Fab fragment.

[0298] In a particular embodiment, the antibody specifically binds to the cancer antigen.

[0299] In a particular embodiment, the antibody specifically binds to the hepatocyte antigen.

[0300] In a particular embodiment, the antibody specifically binds to the antigen presented on the macrophage.

[0301] In certain embodiments, the antibody specifically binds to intact complement or a fragment thereof. In certain embodiments, the antibody specifically binds to one or more immunodominant epitopes within intact complement or a fragment thereof.

[0302] In certain embodiments, the antibody specifically binds to a cell surface receptor. In certain embodiments, the antibody specifically binds to a cell surface receptor ligand.

[0303] In certain embodiments, the antibody specifically binds to epidermal growth factor (EGF) protein, for example, human EGF. In certain embodiments, the antibody specifically binds to one or more immunodominant epitopes within the EGF protein.

[0304] In certain embodiments, the antibody specifically binds to the epidermal growth factor receptor (EGFR) protein, for example, human EGFR. In certain embodiments, the antibody specifically binds to one or more immunodominant epitopes within the EGFR protein. In certain embodiments, the antibody contains a CDR present in cetuximab. In another certain embodiment, the antibody contains a variable light chain and a variable heavy chain present in cetuximab. In certain embodiments, the antibody is cetuximab. In certain embodiments, the antibody contains a CDR present in matsuzumab. In another certain embodiment, the antibody contains a variable light chain and a variable heavy chain present in matsuzumab. In certain embodiments, the antibody is matsuzumab.

[0305] In certain embodiments, the antibody specifically binds to vascular endothelial growth factor (VEGF) proteins, such as human VEGF protein. In certain embodiments, the antibody specifically binds to one or more immunodominant epitopes within the VEGF protein.

[0306] In certain embodiments, the antibody specifically binds to vascular endothelial growth factor receptor (VEGFR) protein, for example, human VEGFR protein. In certain embodiments, the antibody specifically binds to vascular endothelial growth factor receptor 2 (VEGFR2) protein, for example, human VEGFR2 protein. In other specific embodiments, the antibody specifically binds to vascular endothelial growth factor receptor 3 (VEGFR3) protein, for example, human VEGFR3 protein. In certain embodiments, the antibody specifically binds to one or more immunodominant epitopes within the VEGFR protein, VEGFR2 protein, or VEGFR3 protein.

[0307] In certain embodiments, the antibody specifically binds to fibroblast growth factor (FGF), for example, human FGF. In certain embodiments, the antibody specifically binds to one or more immunodominant epitopes within the FGF protein.

[0308] In certain embodiments, the antibody specifically binds to fibroblast growth factor receptor (FGFR), for example, human FGFR. In certain embodiments, the antibody specifically binds to fibroblast growth factor receptor 2 (FGFR2) protein, for example, human FGFR2 protein, for example, FGFR2b protein. In other certain embodiments, the antibody specifically binds to fibroblast growth factor receptor 3 (FGFR3) protein, for example, human FGFR3 protein. In certain embodiments, the antibody specifically binds to one or more immunodominant epitopes within the FGFR protein, FGFR2 protein, or FGFR3 protein. In certain embodiments, the antibody contains CDRs present in bofatamab. In another certain embodiment, the antibody contains variable light chains and variable heavy chains present in bofatamab. In certain embodiments, it is bofatamab. In certain embodiments, the antibody contains CDRs present in bemarituzumab. In another certain embodiment, the antibody contains variable light chains and variable heavy chains present in bemarituzumab. In certain embodiments, it is bemalituzumab.

[0309] In certain embodiments, the antibody specifically binds to the receptor tyrosine kinase cMET protein. In certain embodiments, the antibody specifically binds to one or more immunodominant epitopes within the receptor tyrosine kinase cMET protein. In certain embodiments, the antibody contains a CDR present in onartuzumab (MetMAb; see, e.g., CAS number 133766-06-9). In certain embodiments, the antibody contains variable light and heavy chains present in onartuzumab. In certain embodiments, the antibody is onartuzumab. In certain embodiments, the antibody contains a CDR present in emibetuzumab (LY2875358; see, e.g., CAS number 1365287-97-3). In certain embodiments, the antibody contains variable light and heavy chains present in emibetuzumab. In certain embodiments, the antibody is emibetuzumab. In certain embodiments, the antibody specifically binds to the CD47 protein, e.g., human CD47 protein. In certain embodiments, the antibody specifically binds to one or more immunodominant epitopes within the CD47 protein. In certain embodiments, the antibody contains a CDR present in Hu5F9-G4(5F9). In another certain embodiment, the antibody contains a variable light chain and a variable heavy chain present in Hu5F9-G4(5F9). In certain embodiments, it is Hu5F9-G4(5F9).

[0310] In certain embodiments, the antibody specifically binds to an immune checkpoint inhibitor. In certain embodiments, the antibody binds to one or more immunodominant epitopes within the immune checkpoint inhibitor.

[0311] In certain embodiments, the antibody specifically binds to programmed death proteins, such as human PD-1. In certain embodiments, the antibody specifically binds to one or more immunodominant epitopes within the PD-1 protein. In certain embodiments, the antibody contains CDRs present in nivolumab. In another certain embodiment, the antibody contains variable light and heavy chains present in nivolumab. In certain embodiments, the antibody is nivolumab. In certain embodiments, the antibody contains CDRs present in pembrolizumab. In another certain embodiment, the antibody contains variable light and heavy chains present in pembrolizumab. In certain embodiments, the antibody is pembrolizumab.

[0312] In certain embodiments, the antibody specifically binds to programmed death ligand-1 (PD-L1) protein, for example, human PD-L1. In certain embodiments, the antibody specifically binds to one or more immunodominant epitopes within the PD-L1 protein. In certain embodiments, the antibody contains a CDR present in atezolizumab. In another certain embodiment, the antibody contains a variable light chain and a variable heavy chain present in atezolizumab. In certain embodiments, the antibody is atezolizumab. In certain embodiments, the antibody contains a CDR present in 29E.2A3 (BioXCell). In another certain embodiment, the antibody contains a variable light chain and a variable heavy chain present in 29E.2A3. In certain embodiments, the antibody is 29E.2A3.

[0313] In a particular embodiment, the antibody binds to TIM3. In a particular embodiment, the antibody binds to one or more immunodominant epitopes within TIM3.

[0314] In certain embodiments, the antibody specifically binds to a lectin. In certain embodiments, the antibody specifically binds to one or more immunodominant epitopes within the lectin. In certain embodiments, the antibody binds to SIGLEC. In certain embodiments, the antibody binds to one or more immunodominant epitopes within SIGLEC. In certain embodiments, the antibody binds to a cytokine receptor. In certain embodiments, the antibody binds to one or more immunodominant epitopes within the cytokine receptor. In certain embodiments, the antibody binds to sIL6R. In certain embodiments, the antibody binds to one or more immunodominant epitopes within sIL6R. In certain embodiments, the antibody binds to a cytokine. In certain embodiments, the antibody binds to one or more immunodominant epitopes within the cytokine. In further certain embodiments, the antibody binds to MCP-1, TNF (e.g., TNF alpha), IL1a, IL1b, IL4, IL5, IL6, IL12 / IL23, IL13, IL17, or p40. Furthermore, in certain embodiments, the antibody binds to one or more immunodominant epitopes within MCP-1, TNF (e.g., TNF alpha), IL1a, IL1b, IL4, IL5, IL6, IL12 / IL23, IL13, IL17, or p40.

[0315] In certain embodiments, the antibody binds to major histocompatibility genes (e.g., MHC class I or class II molecules). In certain embodiments, the antibody binds to one or more immunodominant epitopes within major histocompatibility genes (e.g., MHC class I or class II molecules). In certain embodiments, the antibody binds to beta-2 microglobulin. In certain embodiments, the antibody binds to one or more immunodominant epitopes within beta-2 microglobulin.

[0316] Table A shows the heavy and light chain sequences of exemplary anti-EGFR antibodies (see, for example, cetuximab, CAS numbers 205923-56-4). Table A: Heavy chain QVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYYDYEFAYWGQG TLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (Sequence ID 1) Light chain DILLTQSPVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLIKYASESISGIPSRFSGSGGTDFTLSINSVESEDIADYYCQQNNNWPTTFGAGTKLELK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (Sequence 2)

[0317] Table B shows the heavy and light chain sequences of exemplary Fab fragments of anti-EGFR antibodies (see, for example, matsuzumab, NCBI accession numbers 3C09H_H and 3C09_L, CAS number 339186-68-4). Table B: Heavy chain Fab QVQLVQSGAEVKKPGASVKVSCKASGYTFTSHWMHWVRQAPGQGLEWIGEFNPSNGRTNYNEKFKSKATMTVDTSTNTAYMELSSLRSEDTAVYYCASRDYDYAGRYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKS (Sequence ID 3) Light chain DIQMTQSPSSLSASVGDRVTITCSASSSVTYMYWYQQKPGKAPKLLIYDTSNLASGVPSRFSGSGSGTDYTFTISSLQPEDIATYYCQQWSSHIFTFGQGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGE (Sequence No. 4)

[0318] Table C shows the heavy and light chain sequences of exemplary anti-PD-L1 antibodies (see, for example, atezolizumab, CAS number 138723-44-3). Table C: Heavy chain EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGT LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKT HTCPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT ISKAKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (Sequence ID 5) Light chain DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (Sequence ID 6)

[0319] Pharmaceutical composition In another embodiment, a pharmaceutical composition comprising one or more conjugates disclosed herein and a pharmaceutically acceptable carrier is provided herein.

[0320] In certain embodiments, the pharmaceutical compositions provided herein comprise a therapeutically effective amount of one or more of the conjugates provided herein, and optionally one or more additional prophylactic or therapeutic agents, in a pharmaceutically acceptable carrier. The pharmaceutical compositions may be useful for the prevention, treatment, management, or improvement of one or more of the diseases or disorders or symptoms described herein.

[0321] Suitable pharmaceutically acceptable carriers for administering the conjugates provided herein include any such carriers known to those skilled in the art to be suitable for a particular mode of administration.

[0322] The conjugates described herein may be formulated as the sole pharmaceutically active ingredient in a composition, or they may be combined with other active ingredients.

[0323] In certain embodiments, the conjugate is formulated into one or more suitable pharmaceutical formulations, such as a solution in a sterile solution or suspension for parenteral administration, a suspension, a powder, a sustained-release formulation, or an elixir, or into a transdermal patch formulation and a dry powder inhalant formulation.

[0324] In the compositions provided herein, the conjugates described herein may be mixed with a suitable pharmaceutical carrier. The concentration of the conjugate in the composition may be effective, for example, for delivering an amount that, at the time of administration, treats, prevents, or improves the condition or disorder or its symptoms described herein.

[0325] In certain embodiments, the pharmaceutical compositions provided herein are formulated for single-dose administration. To formulate the composition, a weight fraction of the conjugate is dissolved, suspended, dispersed, or otherwise mixed in a selected carrier at an effective concentration such that a treated condition is alleviated, prevented, or one or more symptoms are improved.

[0326] The concentration of the conjugate in the pharmaceutical compositions provided herein will depend, for example, on the physicochemical properties of the conjugate, the administration schedule, and the amount administered, as well as other factors known to those skilled in the art.

[0327] The pharmaceutical compositions described herein are provided for administration to subjects, e.g., humans or animals (e.g., mammals), in unit dosage forms such as sterile parenteral (e.g., intravenous) solutions or suspensions containing a suitable amount of the compound or a pharmaceutically acceptable derivative thereof. The pharmaceutical compositions are also provided for administration to humans and animals in unit dosage forms, including oral or nasal solutions or suspensions and oil-water emulsions containing a suitable amount of the conjugate or a pharmaceutically acceptable derivative thereof. The conjugate is formulated and administered in unit or multiple dosage forms in certain embodiments. As used herein, unit dose forms refer to physically distinct units, individually packaged as known in the art, suitable for human or animal (e.g., mammalian) subjects. Each unit dose contains a predetermined amount of conjugate sufficient to produce the desired therapeutic effect in conjunction with the necessary pharmaceutical carrier, vehicle, or diluent. Examples of unit doses include ampoules and syringes, as well as individually packaged capsules. Unit dose forms can be administered in divisions or in multiples thereof. A multi-dose form is a package containing multiple identical unit doses, administered in separate unit doses. Examples of multi-dose forms include vials, capsule bottles, or bottles. Therefore, in certain embodiments, a multi-dose form is a package containing multiple unit doses that are not separated.

[0328] In certain embodiments, the conjugates of this specification are present in liquid pharmaceutical formulations. Liquid pharmaceutically administered formulations can be prepared, for example, by dissolving, dispersing, or otherwise mixing the conjugate and any pharmaceutical adjuvant in a carrier such as water, saline, aqueous dextrose, glycerol, or glycol to form a solution or suspension. In certain embodiments, the pharmaceutical compositions provided herein to be administered may also contain small amounts of non-toxic adjuvants, such as wetting agents, emulsifiers, solubilizers, and pH buffers.

[0329] Practical methods for preparing such dosage forms are publicly known or will become 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 antibody in the range of 0.005% to 100%, with the remainder consisting of a non-toxic carrier.

[0330] Parenteral administration, in certain embodiments, is characterized by subcutaneous, intramuscular, or intravenous injection, and is intended herein. Injectable preparations can be prepared in conventional forms as liquid solutions or suspensions, in solid forms suitable for solutions or suspensions in liquids prior to injection, or as emulsions. Injectable preparations, solutions, and emulsions also contain one or more excipients. Suitable excipients include, for example, water, saline, dextrose, glycerol, or ethanol. Other routes of administration may include intraenteral, intracerebral, nasal, intra-arterial, intracardiac, intraosseous, intrathecal, and intraperitoneal administration.

[0331] Preparations for parenteral administration include sterile solutions prepared for injection, sterile dried soluble products, lyophilized powders prepared to be combined with a solvent immediately before use, subcutaneous tablets, sterile suspensions prepared for injection, sterile dried insoluble products prepared to be combined with a vehicle immediately before use, and sterile emulsions. The solutions may be aqueous or non-aqueous.

[0332] When administered intravenously, suitable carriers include solutions containing physiological saline or phosphate-buffered saline (PBS), as well as thickeners and solubilizers, such as glucose, polyethylene glycol, and polypropylene glycol, and mixtures thereof.

[0333] Pharmaceutically acceptable carriers used in parenteral formulations include aqueous vehicles, non-aqueous vehicles, antimicrobial agents, isotonic agents, buffers, antioxidants, local anesthetics, suspensions and dispersants, emulsifiers, metal ion sequestering agents or chelating agents, and other pharmaceutically acceptable substances.

[0334] Pharmaceutical carriers also include ethyl alcohol, polyethylene glycol, and propylene glycol for water-miscible vehicles, as well as sodium hydroxide, hydrochloric acid, citric acid, or lactic acid for pH adjustment.

[0335] In certain embodiments, intravenous or intra-arterial infusion of a sterile aqueous solution containing the conjugate described herein is an effective mode of administration. Another embodiment is a sterile aqueous or oily solution or suspension containing the conjugate described herein, which is injected as needed to produce the desired pharmacological effect.

[0336] In certain embodiments, the pharmaceutical formulation is a lyophilized powder, which can be reconstituted for administration as a solution, emulsion, and other mixture. They can also be reconstituted and formulated as a solid or gel.

[0337] Lyophilized powders are prepared by dissolving the conjugates provided herein in a suitable solvent. In some embodiments, the lyophilized powders are sterile. Suitable solvents may contain excipients that improve the stability or other pharmacological properties of the powder or the reconstituted solution prepared from the powder. Possible excipients include, but are not limited to, dextrose, sorbital, fructose, corn syrup, xylitol, glycerin, glucose, sucrose, or other suitable agents. Suitable solvents may also contain buffers such as citrate phosphate, sodium phosphate, or potassium phosphate, or in certain embodiments, other such buffers known to those skilled in the art at a nearly neutral pH. Examples of formulations are provided by subsequent sterile filtration of the solution, followed by lyophilization under standard conditions known to those skilled in the art. In certain embodiments, the resulting solution is distributed into vials for lyophilization. Lyophilized powders can be stored under suitable conditions, for example, at about 4°C to room temperature.

[0338] Reconstitution of this lyophilized powder with distilled water for injection yields a formulation for parenteral administration. For reconstitution, the lyophilized powder is added to sterile water or another suitable carrier.

[0339] In certain embodiments, the conjugates provided herein may be formulated for topical administration or application, for example in the eyes, for example on the skin and mucous membranes, in the form of gels, creams, and lotions, as well as for application to the eyes, or for intracisional or intraspinal application. Topical administration is intended for transdermal delivery, and also for administration to the eyes or mucous membranes, or for inhalation therapy. Nasal solutions containing the active compound alone or in combination with other pharmaceutically acceptable excipients may also be administered.

[0340] Usage and Instructions: In one embodiment, a method for removing a polypeptide of interest (target protein) from the surface of a cell using the conjugate described herein is provided. In another embodiment, a method for removing a polypeptide of interest (target protein) from the extracellular environment using the conjugate described herein is provided. For example, in one embodiment, a method for removing a polypeptide of interest (target protein) from the surface of a cell by sequestering the target protein within the lysosome of a cell using the conjugate described herein is provided. In another embodiment, a method for removing a polypeptide of interest (target protein) from the extracellular space (extracellular environment) of a cell by sequestering the target protein within the lysosome of a cell using the conjugate described herein is provided. In yet another embodiment, a method for removing a polypeptide of interest (target protein) from the surface of a cell by sequestering the target protein within the lysosome of a cell and degrading the target protein using the conjugate described herein is provided. In yet another embodiment, a method for removing a polypeptide of interest (target protein) from the extracellular space (extracellular environment) of a cell by sequestering the target protein within the lysosome of a cell and degrading the target protein using the conjugate described herein is provided.

[0341] Removal of a target protein may refer to a reduction or depletion of the target protein from the cell surface or extracellular space, or a reduction or depletion of the amount of the target protein in the extracellular environment, i.e., on the cell surface or in the extracellular environment. In some embodiments, this method is a method for reducing the amount or level of the target protein in a biological system or cell sample.

[0342] In one embodiment, a method for sequestering a polypeptide of interest (target protein) within a cell's lysosome using the conjugate described herein is provided. In one embodiment, a method for sequestering a polypeptide of interest (target protein) within a cell's lysosome and degrading the polypeptide of interest is provided using the conjugate described herein.

[0343] In one embodiment, a method for degrading a polypeptide of interest (target protein) using a conjugate described herein is provided herein.

[0344] In one embodiment, a method for depleting a polypeptide of interest described herein (target protein) by degradation via the cellular lysosomal pathway is provided herein.

[0345] In another embodiment, a method is provided herein for depleting a polypeptide of interest (target protein) described herein by administering an effective amount of a conjugate or pharmaceutically acceptable salt, or a pharmaceutically acceptable composition, described herein, to a subject requiring it. In certain embodiments, the subject is a mammal (e.g., human).

[0346] In certain embodiments, the target protein is a membrane-bound protein. In certain embodiments, the target protein is an extracellular protein.

[0347] In certain embodiments, the target protein is a VEGF protein, an EGFR protein, a VEGFR protein, a PD-L1 protein, a FGFR2 protein, or a FGFR3 protein.

[0348] In another embodiment, a method for treating a disease or disorder is provided herein by administering an effective amount of the conjugate or pharmaceutically acceptable salt or pharmaceutically acceptable composition described herein to a subject, for example, a human being in need thereof.

[0349] The terms “administer,” “administration,” or “administering” refer to the act of injecting or otherwise physically delivering a substance (e.g., a conjugate or pharmaceutical composition provided herein) into a subject or patient (e.g., a human) by, for example, mucosal, topical, intradermal, parenteral, intravenous, intramuscular delivery, and / or any other method of physical delivery described herein or known in the art. In certain embodiments, administration is by intravenous infusion.

[0350] The terms “effective dose” or “therapeutic effective dose” refer to an amount of therapeutic agent (e.g., a conjugate or pharmaceutical composition provided herein) sufficient to treat, diagnose, prevent, delay the onset of, reduce the severity and / or duration of, and / or improve, a given condition, disorder or disease and / or symptoms associated therewith. These terms also encompass amounts necessary to mitigate, slow, or improve the progression or advancement of a given disease, mitigate, slow, or improve the recurrence, onset or initiation of a given disease, and / or improve or enhance the preventive or therapeutic effect of another treatment, or to act as a bridge to another treatment. In some embodiments, “effective dose” as used herein also refers to an amount of the conjugate described herein to achieve a particular result.

[0351] In a particular embodiment, if the disorder or disease is cancer, “effective dose” or “therapeutic effective dose” means the amount of the conjugate or pharmaceutical composition provided herein that, when administered to a person having cancer, is sufficient to treat cancer. “Treatment” or “treatment” of cancer includes one or more of the following: (1) To limit / inhibit the growth of cancer, for example, to limit its development. (2) Reducing / preventing the spread of cancer, for example, reducing / preventing metastasis, (3) To reduce cancer, for example, to induce cancer regression, (4) Reducing / preventing cancer recurrence, (5) To alleviate the symptoms of cancer.

[0352] The terms “subject” and “patient” are used interchangeably. The subject may be a non-primate (e.g., cattle, pigs, horses, cats, dogs, goats, rabbits, rats, mice, etc.) or a primate (e.g., monkeys and humans), such as a mammal, such as a human. In certain embodiments, the subject is a mammal diagnosed with the disease or disorder provided herein, such as a human. In other embodiments, the subject is a mammal at risk of developing the disease or disorder provided herein, such as a human. In certain embodiments, the subject is a human.

[0353] The term “treatment” may refer to any protocol, method, composition, formulation, and / or agent that can be used to prevent, treat, manage, or improve a disease or disorder or its symptoms (e.g., a disease or disorder provided herein, or one or more symptoms or conditions associated therewith). In certain embodiments, the term “treatment” may refer to pharmacotherapy, adjunctive therapy, radiation, surgery, biological therapy, supportive therapy, and / or other treatments useful for treating, managing, preventing, or improving a disease or disorder or one or more symptoms therewith. In certain embodiments, the term “treatment” may refer to treatments other than the conjugate or its pharmaceutical composition described herein.

[0354] In a particular embodiment, a disease or disorder is treated by depletion of a target protein through degradation via the lysosomal pathway.

[0355] In certain embodiments, a disease or disorder is treated by depletion of certain proteins, e.g., soluble proteins, e.g., secreted proteins, cell surface proteins (e.g., cell surface receptor proteins, e.g., tyrosine kinase receptors, soluble cytokine receptors, and immune checkpoint receptors, e.g., EGFR, VEGFR, FGFR, and PD-L1), lectins, complement, lipoproteins, transport proteins, MHC class I and class II molecules, cytokines, chemokines, and / or receptors, or any of the aforementioned fragments or subunits.

[0356] In a particular embodiment, the disease or disorder is cancer.

[0357] In a particular embodiment, cancer is selected from the group consisting of bladder cancer, breast cancer, cervical cancer, bile duct cancer, endometrial cancer, hepatocellular carcinoma, kidney cancer, melanoma, myeloid neoplasm, non-small cell lung cancer (NSCLC), Ewing's sarcoma, and Hodgkin lymphoma.

[0358] In certain embodiments, cancer is a solid tumor.

[0359] In certain embodiments, the disease or disorder is an inflammatory or autoimmune disease.

[0360] In a particular embodiment, the disease or disorder is an inflammatory disease.

[0361] In certain embodiments, the disease or disorder is an autoimmune disease.

[0362] definition The terms “protein” and “polypeptide” are used interchangeably. Proteins may contain non-amino acid portions (which may be glycoproteins, for example) and / or may be otherwise processed or modified. Those skilled in the art will understand that a “protein” may be a complete protein chain (with or without a signal sequence) or a portion thereof, as produced by a cell. Those skilled in the art will understand that a protein may contain more than one protein chain, linked, for example, non-covalently or covalently, by one or more disulfide bonds or by other means. Polypeptides may contain L-amino acids, D-amino acids, or both, and may contain any of the various amino acid modifications or analogs known in the art. Useful modifications include, for example, terminal acetylation, amidation, methylation, etc. In some embodiments, a protein may contain native amino acids, non-native amino acids, synthetic amino acids, and combinations thereof. In some embodiments, a protein may be an antibody, an antibody fragment, a biologically active portion thereof, and / or a characteristic portion thereof.

[0363] The terms “antibody” and “immunoglobulin” are terms of the art and may be used interchangeably herein to refer to molecules having an antigen-binding site that specifically binds to an antigen.

[0364] In certain embodiments, an isolated antibody (e.g., a monoclonal antibody) or an antigen-binding fragment of a protein of interest, such as EGFR, which specifically binds to EGFR, is conjugated, for example, to one or more lysosomal targeting moieties via a linker.

[0365] An "antigen" is a portion or molecule containing an epitope to which an antibody can specifically bind. Therefore, an antigen is also specifically bound by an antibody. In certain embodiments, the antigen to which the antibody described herein binds is a protein of interest, for example, EGFR (e.g., human EGFR), or a fragment thereof, or for example, the extracellular domain of EGFR (e.g., human EGFR).

[0366] "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 adjacent amino acids, or it may consist of amino acids from two or more non-adjacent regions of the antigen.

[0367] In the context of antibody binding, the terms “binds,” “binds to,” “specifically binds,” or “specifically binds to” refer to antibodies that bind to an antigen (e.g., an epitope) in such a way that such binding is understood by those skilled in the art. For example, a molecule that specifically binds to an antigen may bind to other polypeptides with a lower affinity, generally determined by, for example, immunoassays, Biacore®, KinExA 3000 instrument (Sapidyne Instruments, Boise, ID) or other assays known in the art. In a particular embodiment, a molecule that specifically binds to an antigen may bind to another antigen, such as K d Affinity (K) is lower than at least 2-log, 2.5-log, 3-log, and 4-log than the affinity (K) of the two-log, 2.5-log, 3-log, and 4-log scales. d It binds to an antigen having (higher affinity). In another specific embodiment, the molecule that specifically binds to the antigen does not cross-react with other proteins. In another specific embodiment, where EGFR is the protein of interest, the molecule that specifically binds to the antigen does not cross-react with other non-EGFR proteins.

[0368] Antibodies may include, for example, monoclonal antibodies, recombinant antibodies, monospecific antibodies, multispecific antibodies (including bispecific antibodies), human antibodies, humanized antibodies, chimeric antibodies, synthetic antibodies, tetrameric antibodies containing two heavy chains and two light chain molecules, antibody light chain monomers, antibody heavy chain monomers, antibody light chain dimers, antibody heavy chain dimers, antibody light chain / antibody heavy chain pairs, antibodies having two light chain / heavy chain pairs (e.g., identical pairs), intrabodies, heteroconjugate antibodies, single-domain antibodies, monovalent antibodies, bivalent antibodies (including monospecific or bispecific bivalent antibodies), single-chain antibodies, or single-chain Fv(scFv), camelized antibodies, aphibodies, Fab fragments, F(ab') fragments, F(ab')2 fragments, disulfide-linked Fv(sdFv), anti-idiotype (anti-Id) antibodies (e.g., anti-anti-Id antibodies), and any of the above epitope-linked fragments.

[0369] The antibody may be any type of immunoglobulin molecule (e.g., IgG, IgE, IgM, IgD, IgA, or IgY), any class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, or IgA2), or any subclass (e.g., IgG2a or IgG2b). In certain embodiments, the antibody described herein is an IgG antibody (e.g., human IgG), or a class (e.g., human IgG1, IgG2, IgG3, or IgG4), or a subclass thereof.

[0370] In certain embodiments, the antibody is a four-chain antibody unit comprising two heavy (H) chain / light (L) chain pairs, where 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 include a constant region, e.g., a human constant region. 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 yet another specific embodiment, the H chain constant region of such an antibody includes a gamma heavy chain constant region, e.g., a human gamma heavy chain constant region. In certain embodiments, such an antibody includes an IgG constant region, e.g., a human IgG constant region.

[0371] The terms “constant region” or “constant domain” are well-known antibody terms in the relevant art (sometimes referred to as “Fc”) and refer to the light and / or heavy chain carboxyl-terminal portions of the antibody moiety, for example, those that do not directly participate in the binding of the antibody to an antigen but can exhibit various effector functions, such as interaction with Fc receptors. These terms refer to parts of immunoglobulin molecules that generally have more conserved amino acid sequences compared to immunoglobulin variable domains.

[0372] When used in reference to antibodies, the term "heavy chain" can refer to any different types that give rise to the IgA, IgD, IgE, IgG, and IgM classes of antibodies, such as alpha (α), delta (δ), epsilon (ε), gamma (γ), and mu (μ), including the IgG subclasses IgG1, IgG2, IgG3, and IgG4, respectively, based on the amino acid sequence of the constant domain.

[0373] When used in relation to antibodies, the term "light chain" can refer to any different type, for example, a lambda (λ) kappa (κ) light chain based on the amino acid sequence of its constant domain. Light chain amino acid sequences are well known in the art. In certain embodiments, the light chain is a human light chain.

[0374] The term “monoclonal antibody” is a well-known technical term referring to an antibody obtained from a homogeneous or substantially homogeneous population of antibodies. The term “monoclonal” is not limited to any particular method for producing the antibody. Generally, a population of monoclonal antibodies can be produced by a cell, a population of cells, or a cell line. In certain embodiments, “monoclonal antibody” as used herein is an antibody produced by a single cell (e.g., a hybridoma or a host cell producing recombinant antibodies) and the antibody binds specifically to an epitope as determined, for example, by ELISA or other antigen-binding or competitive binding assays known in the art or in the examples provided herein. In certain embodiments, the monoclonal antibody may be a chimeric antibody or a humanized antibody. In certain embodiments, the monoclonal antibody may be a monovalent or polyvalent (e.g., bivalent) antibody. In certain embodiments, the monoclonal antibody may be a monospecific or multispecific (e.g., bispecific) antibody.

[0375] The term "variable region" or "variable domain" refers to a portion of an antibody, generally a portion of the light or heavy chain, typically the approximately 110–120 amino acids from the amino terminus to the mature heavy chain and approximately 90–100 amino acids from the mature light chain. The variable region includes a complementarity-determining region (CDR) adjacent to the framework region (FR). Generally, the spatial positioning of the CDR and FR in the N-terminus to C-terminus direction is as follows: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4. While we do not wish to be bound by any particular mechanism or theory, the CDRs of the light and heavy chains are thought to be primarily involved in antibody-antigen interaction and antibody specificity to epitopes. In certain embodiments, the numbering of amino acid positions of the antibodies described herein follows the EU index, as in Kabat et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, USD Department of Health and Human Services, NIH Publication No. 91-3242. In certain embodiments, the variable region is the human variable region.

[0376] In certain embodiments, the CDRs of antibodies may be (i) numbered using 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, USD Department of Health and Human Services, NIH Publication No. 91-3242), or (ii) numbered using the Chothia numbering scheme, referred to herein 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.) 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 See Variable Domains, in Antibody Engineering, Kontermann and Dubel, eds., Chapter 31, pp. 422-439, Springer-Verlag, Berlin (2001), or (iii) the ImMunoGeneTics (IMGT) numbering system, as described, for example, in Lefranc, 1999, The Immunologist, 7:132-136 and Lefranc et al., 1999, Nucleic Acids Res., 27:209-212 ("IMGT CDRs"), or (iv) the AbM numbering system, as referred to herein as "AbM CDR", as described, for example, in MacCallum et al., 1996, J.Mol.Biol., 262: 732-745. See also, for example, Martin, A., “Protein Sequence and Structure Analysis of Antibody Variable Domains,” in Antibody Engineering, Kontermann and Dubel, eds., Chapter 31, pp. 422-439, Springer-Verlag, Berlin (2001), or (v) the contact numbering system referred to herein as “contact CDR” (the contact definition is based on the analysis of available complex crystal structures (bioinf.org.uk / abs)) (see, for example, MacCallum et al., 1996, J.Mol.Biol., 262:732-745).

[0377] The terms “full-length antibody,” “intact antibody,” and “whole antibody” are used interchangeably herein to refer to an antibody in substantially intact form and not to an antibody fragment as defined below. These terms, in particular, refer to an antibody having a heavy chain containing an Fc region.

[0378] An "antibody fragment" comprises only a portion of an intact antibody, the portion retaining at least one, two, three, and almost all of the functions normally associated with that portion when present in an intact antibody. In one embodiment, the antibody fragment contains the antigen-binding site of an intact antibody and therefore retains the ability to bind to an antigen. In another embodiment, an antibody fragment, such as an antibody fragment containing an Fc region, retains at least one of the biological functions normally associated with the Fc region when present in an intact antibody. Such functions may include FcRn binding, antibody half-life regulation, conjugate function, and complement binding. In yet another embodiment, the antibody fragment is a monovalent antibody having substantially the same in vivo half-life as an intact antibody. For example, such an antibody fragment may contain an antigen-binding arm linked to an Fc sequence that can give the fragment in vivo stability.

[0379] "Alkyl" means a linear or branched saturated hydrocarbon group containing 1 to 10 carbon atoms, and in certain embodiments, containing 1 to 6 carbon atoms. In certain embodiments, alkyl is a group containing 1 to 4 carbon atoms ("C 1-4 Alkyl) contains 1 to 3 carbon atoms ("C"). In certain embodiments, alkyl contains 1 to 3 carbon atoms ("C"). 1-3 The alkyl group includes methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylhexyl, n-heptyl, n-octyl, n-nonyl, or n-decyl.

[0380] "Alkyl" refers to a linear or branched saturated divalent hydrocarbon group containing 1 to 10 carbon atoms. In certain embodiments, alkylene is a group containing 1 to 6 carbon atoms ("C 1-6 Contains alkylene.

[0381] "Halo" refers to a fluoro, chloro, bromo, or iodine group.

[0382] "CN" stands for cyano group.

[0383] Unless otherwise specified, if a compound can take on alternative tautomers, positional isomers, and / or stereoisomers, all alternative isomers are intended to be included within the scope of the claimed subject matter. For example, if a compound is described as having a particular optical isomer D- or L-, both optical isomers are intended to be included herein. For example, if a compound is described as having one of two tautomer forms, both tautomers are intended to be included herein. Thus, the compounds provided herein may be enantiomerically pure or may be stereoisomers or mixtures of diastereomers. The compounds provided herein may contain a chiral center. Such a chiral center may be in either an (R) or (S) configuration, or a mixture thereof. The chiral centers of the compounds provided herein may undergo epimerization in vivo. Thus, those skilled in the art will recognize that a dose of the compound in its (R) form is equivalent to a dose of the compound in its (S) form with respect to a compound that undergoes epimerization in vivo.

[0384] This disclosure also encompasses all suitable isotopic variants of the compounds according to this disclosure, whether radioactive or not. An isotopic variant of a compound according to this disclosure is understood to mean a compound in which at least one atom in the compound according to this disclosure is replaced with another atom having the same atomic number but a different atomic mass than that normally or predominantly found in nature. Examples of isotopes that can be incorporated into the compounds according to this disclosure are 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 131 It is an isotope of I. Certain isotopic variants of the compounds according to this disclosure, in particular isotopic variants incorporating one or more radioactive isotopes, may be useful, for example, for testing the mechanism of action or the distribution of active compounds in the body. 3 H, 14 C, and / or 18 Compounds labeled with 1F isotopes are suitable for this purpose. In addition, the incorporation of deuterium isotopes, for example, can result in certain therapeutic benefits as a result of greater metabolic stability of the compound, such as an extended half-life in the body or a reduction in the required active dose. In some embodiments, hydrogen atoms in the compounds described herein can be replaced with deuterium atoms. In certain embodiments, “deuterated” as applied to a chemical group refers to a chemical group in which a substantially greater amount of deuterium than its natural abundance is isotope-enriched, unless otherwise indicated. Isotope variants of the compounds according to this disclosure can be prepared in various ways, for example, by using certain reagents and / or corresponding isotope modifications of the starting compounds in the methods and examples described below.

[0385] Therefore, any of the embodiments described herein means that include salts, single stereoisomers, mixtures of stereoisomers, and / or isotopic forms of the compound.

[0386] Unless otherwise specified, the terms “about” or “approximately” mean an acceptable error of a particular value as determined by those skilled in the art, which depends in part on how that value is measured or determined. In certain embodiments, the terms “about” or “approximately” mean a standard deviation of 1, 2, or 3. In certain embodiments, the terms “about” or “approximately” mean within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.25%, 0.2%, 0.1%, or 0.05% of a given value or range. In certain embodiments where an integer is required, the term “about” means within plus or minus 10% of a given value or range, rounded up or down to the nearest integer.

[0387] In the event of any discrepancy between a chemical name and a chemical structure in this specification, the chemical structure shall prevail.

[0388] Additional Embodiments The nature of this disclosure is described in the following clauses.

[0389] Clause 1. Cell surface mannose-6-phosphate receptor (M6PR) binding compound of formula (XI): [ka] or a salt thereof, During the ceremony, Each W is independently a hydrophilic head group. each Z 1 However, independently, they are selected from arbitrarily substituted (C1-C3) alkylenes and arbitrarily substituted ethenylenes. each Z 2 However, independently, O, S, NR 21 , and C(R 22 ) Selected from 2, each R 21 However, each R is independently selected from H and optionally substituted (C1-C6) alkyl groups. 22However, independently selected from H, halogens (e.g., F), and optionally substituted (C1-C6) alkyl groups, Each Ar is independently an optionally substituted aryl or heteroaryl linking moiety (e.g., an optionally substituted monocyclic or bicyclic aryl or heteroaryl), each Z 3 However, it is an independent connecting part, n is between 1 and 500. L is the linker, Y is the part of interest, If m is 1 and Ar is phenyl, then i) L contains a backbone of at least 16 consecutive atoms, ii) Y is a biomolecule, and / or ii) Z 3 However, it is a compound that is an amide, sulfonamide, urea, or thiourea.

[0390] Clause 2. The compound according to Clause 1, wherein each Ar is independently selected from optionally substituted phenyl, optionally substituted pyridyl, optionally substituted biphenyl, optionally substituted naphthalene, optionally substituted triazole, and optionally substituted phenylene-triazole.

[0391] Clause 3. The compound according to Clause 2, wherein Ar is selected from optionally substituted 1,4-phenylene, optionally substituted 1,3-phenylene, or optionally substituted 2,5-pyridylene.

[0392] Clause 4. If the compound is a compound of formula (XIIa) or (XIIb): [ka] or a salt thereof, During the ceremony, Each R 11 ~R 14 However, independently, 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 Selected from, Each R 25 The compound according to Clause 3, independently selected from H and optionally substituted (C1-C6) alkyl groups.

[0393] Clause 5. The compound according to Clause 1, wherein Ar is optionally substituted with a condensed bicyclic aryl or condensed bicyclic heteroaryl.

[0394] Clause 6. The compound according to Clause 5, wherein Ar is optionally substituted naphthalene or optionally substituted quinoline.

[0395] Clause 7. If the compound is a compound of formula (XIIIa) or (XIIIb): [ka] or a salt thereof, During the ceremony, Each R 11 and R 13 ~R 14 However, independently, 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 Selected from, s is between 0 and 3. Each R 25 The compound according to Clause 6, independently selected from H and optionally substituted (C1-C6) alkyl groups.

[0396] Clause 8. The compound is one of the compounds of formula (XIIIc) to (XIIIh): [ka] The compound described in Clause 7, or a salt thereof.

[0397] Clause 9. Ar is an optionally substituted bicyclic aryl or optionally substituted bicyclic heteroaryl, and the compound is a compound of formula (XIVa): [ka] or a salt thereof, During the ceremony, Each Cy is independently a monocyclic aryl or monocyclic heteroaryl. Each R 11 ~R 15 However, independently, 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 Selected from, s is between 0 and 4, Each R 25 The compound according to Clause 1, independently selected from H and optionally substituted (C1-C6) alkyl groups.

[0398] Clause 10. Ar is an optionally substituted biphenyl, Cy is an optionally substituted phenyl, and the compound is a compound of formula (XIVb): [ka] The compound described in Clause 9, or a salt thereof.

[0399] Clause 11. If the compound is a compound of formula (XIVc) or (XIVd): [ka] The compound described in Clause 10, or a salt thereof.

[0400] Clause 12. A compound according to any one of Clauses 1 to 10, wherein Ar is substituted with at least one OH substituent.

[0401] Clause 13.R 11 ~R 15 A compound according to any one of the clauses 4, 6, 7, 9, and 10, wherein each is H.

[0402] Clause 14.R 11 ~R 15 A compound according to any one of the clauses 4, 6, 7, 9, and 10, wherein at least one of them is an OH group (for example, at least two are OH groups).

[0403] Clause 15.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 )- Selected from, X 1 and X 2 However, O, S, and NR 23 Selected from, R 23 and R 24 However, independently, H, C (1-3) -alkyl (e.g., methyl), and substituted C (1-3) - A compound selected from alkyl groups, as described in any one of clauses 1 to 14.

[0404] Clause 16.Z 3 However, the following applies: [ka] During the ceremony, X1 However, it is either O or S, t is 0 or 1, Each R 23 However, independently, H, C (1-3) -alkyl (e.g., methyl), and substituted C (1-3) - A compound selected from alkyl groups, as described in any one of clauses 1 to 15.

[0405] Clause 17.Z 3 However, -NHC(=X 1 )NH- and X 1 The compound described in Clause 16, wherein the compound is O or S.

[0406] Clause 18. A compound according to any one of Clauses 1 to 14, wherein Ar is a triazole and the compound is a compound of formula (XIIc) or (XIId). [ka]

[0407] Clause 19.Z 3 However, it is an arbitrarily substituted triazole, and the compound is a compound of formula (XIIc) or (XIId): [ka] or a salt thereof, During the ceremony, Each R 11 ~R 14 However, independently, 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 Selected from, Each R 25 The compounds described in Clause 18, independently selected from H and optionally substituted (C1-C6) alkyl groups.

[0408] Article 20.-Ar-Z 3 -A compound as described in any one of clauses 1 to 19, selected from the following: [ka] [ka]

[0409] The compound according to any one of the clauses 1 to 20, wherein m is at least 2 and L is a branched linker that covalently bonds each Ar group to Y.

[0410] The compound described in Clause 21, where m is 2 to 20 (for example, m is 2 to 6, such as 2 or 3).

[0411] Clause 23.m is 20-500 (for example, 20-400, 20-300, or 20-200, or 50-500, or 100-500), L is an α-amino acid polymer (e.g., poly-L-lysine), and there are numerous -Ar-Z 3 - The compound according to Clause 21, wherein the group is covalently bonded to the polymer skeleton via a side chain group (for example, via conjugation of a lysine residue to a side chain amino group).

[0412] Clause 24.m is at least 2, and each Z 3 The linkage portion, via the linker L, is connected to all other Z by at least 16 consecutive atomic chains (for example, by at least 20, at least 25, or at least 30 consecutive atomic chains, and optionally by up to 100 consecutive atomic chains). 3 A compound described in any one of clauses 21 to 23, separated from the binding portion.

[0413] Clause 25. Compound is a compound of formula (XV): [ka] or a salt thereof, During the ceremony, n is between 1 and 500 (for example, n is between 1 and 20, 1 and 10, 1 and 6, or 1 and 5), Each L 1 ~L 7 However, independently, n Z 2 A linking portion that provides a linear or branched linker together between the base and Y, -(L 1 ) a - comprises a linking moiety Ar which is an optionally substituted aryl or heteroaryl group, a is either 1 or 2, A compound according to any one of clauses 1 to 24, wherein b, c, d, e, f, and g are each independently 0, 1, or 2.

[0414] Clause 26. A linear or branched linker is formed by a chain of at least 16 consecutive atoms (e.g., at least 20 consecutive atoms, at least 30 consecutive atoms, or 16 to 100 consecutive atoms) in each Z 2 The compound described in Clause 25, which separates Y.

[0415] The compound described in Clause 25 or 26, wherein Clause 27.n is 1 to 20.

[0416] A compound according to any one of the clauses 25 to 27, wherein n is at least 2 (for example, n is 2 or 3).

[0417] Clause 29.d is >0, L 4 However, it is a branched connecting section, and each L 1 A compound as described in Clause 28, which is covalently bonded to the bonding portion.

[0418] Clause 30. The compound is a compound of formula (XVIa), [ka] During the ceremony, Ar is an optionally substituted aryl or heteroaryl group (e.g., a monocyclic, bicyclic, or tricyclic aryl or heteroaryl group), Z 11 However, the connecting part (for example, a covalent bond, a heteroatom, a group having a skeleton of 1 to 3 atoms in length, or a triazole) r is 0 or 1, A compound according to any one of clauses 25 to 29, wherein n is 1 to 6.

[0419] Clause 31. The compound according to Clause 30, wherein Ar is selected from optionally substituted phenyl, optionally substituted pyridyl, optionally substituted biphenyl, optionally substituted naphthalene, optionally substituted quinoline, optionally substituted triazole, optionally substituted phenyl-triazole, optionally substituted biphenyl-triazole, and optionally substituted naphthalene-triazole.

[0420] Clause 32. The compound according to Clause 31, wherein Ar is optionally substituted 1,4-phenylene.

[0421] Clause 33. The compound according to any one of Clauses 30 to 32, wherein Ar is substituted with at least one hydroxyl group.

[0422] Clause 34.L 1 or -Ar-(Z 11 ) r -The following options are selected: [ka] During the ceremony, Cy is a monocyclic aryl or heteroaryl compound. r is 0 or 1, s is between 0 and 4, R 11 ~R 14 and each R 15However, independently, 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 Selected from, each R 25 However, independently, H, C (1-6) -alkyl and substituted C (1-6) - Selected from alkyl groups, Z 11 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 -, and selected from optionally substituted triazoles, X 1 and X 2 However, O, S, and NR 23 Selected from, R 23 and R 24 However, independently, H, C (1-3) -alkyl (e.g., methyl), and substituted C (1-3) - Compounds selected from alkyl groups, as described in any one of clauses 25 to 33.

[0423] Clause 35.L 1 However, the compounds described in Article 34 are as follows: [ka]

[0424] Clause 36.L 1 However, the compounds described in Article 34 are as follows: [ka]

[0425] Clause 37.L 1 However, the compounds described in Clause 34 are selected from the following: [ka]

[0426] A compound described in any one of clauses 34 to 37, wherein clause 38.r is 0.

[0427] Clause 39.r is 1, Z 11 However, -O-, -NR 23 -, -NR 23 CO-, CONR 23 -, -NR 23 CO2-, -OCONR 23 -, -NR 23 C(=X 1 )NR 23 -, -CR 24 =N- and -CR 24 =NX 2 - Select from, X 1 and X 2 However, O, S, and NR 23 Selected from, each R 23 and R 24 However, independently, H, C (1-3) -alkyl (e.g., methyl), and substituted C (1-3) - Compounds selected from alkyl groups, as described in any one of clauses 34 to 37.

[0428] Clause 40.r is 1, Z 11 However, the following applies: [ka] During the ceremony, X 1 However, it is either O or S, t is 0 or 1, Each R 23 However, independently, H, C (1-3) -alkyl (e.g., methyl), and substituted C(1-3) - Compounds selected from alkyl groups, as described in any one of clauses 34 to 37.

[0429] Clause 41.Z 11 However, -NHC(=X 1 )NH- and X 1 The compound described in Clause 40, wherein the compound is O or S.

[0430] Clause 42.r is 1, Z 11 However, the compound is a triazole, as described in any one of clauses 34 to 37.

[0431] A compound according to any one of Clauses 1 to 42, wherein Y is selected from small molecules, dyes, fluorophores, monosaccharides, disaccharides, trisaccharides, and chemoselective ligation groups, or precursors thereof.

[0432] Clause 44.Y is a biomolecule, and is a compound as described in any one of Clauses 1 to 42.

[0433] Clause 45. The compound described in Clause 44, wherein the biomolecule is selected from peptides, proteins, polynucleotides, polysaccharides, glycoproteins, lipids, enzymes, antibodies, and antibody fragments.

[0434] A compound according to any one of Clauses 1 to 45, wherein Clause 46.Y is a portion that specifically binds to a target protein.

[0435] Clause 47. The compound described in Clause 46, wherein the target protein is a membrane-bound protein.

[0436] Clause 48. The compound described in Clause 46, wherein the target protein is an extracellular protein.

[0437] Clause 49.Y is a compound according to any one of Clauses 46 to 49, selected from antibodies, antibody fragments (e.g., antigen-binding fragments of antibodies), chimeric fusion proteins, engineered protein domains, D protein binders for target proteins, aptamers, peptides, enzyme substrates, and small molecule inhibitors or ligands.

[0438] Clause 50.Y is an antibody or antibody fragment that specifically binds to a target protein, and the compound is a compound of formula (Va): [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, n is between 1 and 20. m is the average load from 1 to 80. Ab is an antibody or antibody fragment that specifically binds to a target protein. The compound according to Clause 49, wherein Z is a residue portion resulting from the covalent bonding of a chemoselective ligation group to a compatible group of Ab.

[0439] The compound described in Clause 49, wherein Clause 51.Y is a small molecule inhibitor or ligand of the target protein.

[0440] Clause 52. The hydrophilic head group W is -OH, -CR 2 R 2 OH, -OP=O(OH)2, -SP=O(OH)2, -NR 3 P=O(OH)2, -OP=O(SH)(OH), -SP=O(SH)(OH), -OP=S(OH)2, -OP=O(N(R 3 )2)(OH), -OP=O(R 3 )(OH), -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, -CONHR3 ,-CONR 3 R 4 -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 -SO2NH2, -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 their salts, selected from During the ceremony, R 1 and R 2 However, independently, hydrogen, SR 3 , halo, or CN, R 3 and R 4 However, independently, H, C 1-6 Alkyl or substituted C 1-6 It is an alkyl group (e.g., -CF3 or -CH2CF3), A, B, and C are each independently CH or N. A compound according to any one of clauses 1 to 51, wherein D is independently either O or S.

[0441] Clause 53. A compound according to Clause 52, wherein W is selected from -P=O(OH)2, -SO3H, -COOH, and -CH(COOH)2, or salts thereof.

[0442] Clause 54.Z 1 However, -(CH2)j -or-(C(R 22 )2) j - and each R 22 However, independently selected from H, halogens (e.g., F), and optionally substituted (C1-C6) alkyl groups, A compound as described in any one of clauses 1 to 53, wherein j is 1 to 3.

[0443] Clause 55.Z 1 However, a compound as described in any one of clauses 1 to 53, wherein the compound is -CH=CH-.

[0444] Clause 56.Z 2 A compound as described in any one of clauses 1 to 55, wherein the compound is O or S.

[0445] Clause 57.Z 2 However, -NR 21 -A compound as described in any one of the clauses 1 to 55.

[0446] Clause 58.Z 2 However, -C(R 22 )2-, and each R 22 A compound according to any one of clauses 1 to 55, independently selected from H, a halogen (e.g., F), and an optionally substituted (C1-C6) alkyl group.

[0447] Clause 59.Z 1 However, -(CH2) j -, selected from substituted (C1-C3) alkylenes and -CH=CH-, j is 1 to 3, Z 2 However, a compound selected from O and CH2, as described in any one of clauses 1 to 53.

[0448] Article 60. Z 1 However, these are -(CH2)2-, -CH2-CF2-, or -CH2-CHF-, Z 2 However, the compound described in clause 60 is O.

[0449] Clause 61.Z 1 However, these are -(CH2)2-, -CH2-CF2-, or -CH2-CHF-, Z 2 However, the compound is CH2, as described in Clause 60.

[0450] Clause 62.Z 1 However, -CH=CH-, Z 2 However, the compound described in clause 60 is O.

[0451] Clause 63.Z 1 However, -CH=CH-, Z 2 However, the compound is CH2, as described in Clause 60.

[0452] Clause 64.X is a compound described in any one of Clauses 1 to 63, selected from the following: [ka]

[0453] Clause 65.n is 1 to 6 (for example, n is 1 to 5, or 2 to 6, or 1, 2, or 3), If d is 0, then n is 1, If d is 1, then n is between 1 and 3. If d is 2, then n is 1 to 6, the compound according to any one of clauses 25 to 64.

[0454] Article 66. Each L 2 However, independently, -C 1-6 -Alkilen-, -NHCO-C 1-6 -Alkilen-, -CONH-C 1-6 -Alkylene-, -O(CH2) p -, and -(OCH2CH2) p - is selected from, and p is between 1 and 10. Each L 3 However, they became independent, [ka] , and -(OCH2CH2) q - A compound according to any one of clauses 25 to 65, selected from, where q is 1 to 10, u is 0 to 10, and w is 1 to 10.

[0455] If clause 67.n is 2 or more, then at least one L 4 However, a compound described in any one of clauses 25 to 66, which is a branched linkage portion that exists.

[0456] Clause 68. Each L 4 However, they became independent, -OCH2CH2-, [ka] , selected from, A compound according to any one of the clauses 25 to 67, wherein each x and y is independently 1 to 10.

[0457] Article 69. Each L 5 However, it became independent, -NHCO-C 1-6 -Alkilen-, -CONH-C 1-6 -Alkilen-, -C 1-6 -Alkilen-, [ka] , or -(OCH2CH2) r -and, Each L 6 However, it became independent, -NHCO-C 1-6 -Alkilen-, -CONH-C 1-6 -Alkilen-, -C 1-6 -Alkylene-, or -(OCH2CH2)- s -and, Each L 7 However, it became independent, -NHCO-C 1-6 -Alkilen-, -CONH-C 1-6 -Alkilen-, -C 1-6 -Alkilen-, -(OCH2CH2) t -, or -OCH2- A compound according to any one of the clauses 25 to 68, wherein r, s, and t are each independently 1 to 20.

[0458] A compound described in any one of the clauses 25 to 69, wherein clause 70.a is 1.

[0459] A compound described in any one of Clauses 25 to 70, wherein at least one of Clauses 71.b, c, e, f, and g is not 0.

[0460] A compound described in any one of clauses 25 to 71, wherein at least one of clauses 72.b or c is not 0, and at least one of e, f, and g is not 0.

[0461] A compound as described in any one of the clauses 25 to 72, wherein clause 73.a, b, and c are each independently 1 or 2.

[0462] Clause 74. A compound according to any one of Clauses 1 to 73, wherein linker L is selected from any one of the structures in Tables 2 to 3.

[0463] Clause 75. A compound selected from the compounds in Tables 5-9, as described in any one of Clauses 1-74.

[0464] Clause 76. Cell surface receptor-binding conjugate of formula (I): [ka] or a salt thereof, During the ceremony, X is a portion that binds to the asialoclycoprotein receptor (ASGPR) on the cell surface, or to the mannose-6-phosphate receptor (M6PR) on the cell surface. n is between 1 and 500 (for example, n is between 1 and 20, 1 and 10, 1 and 6, or 1 and 5), L is the linker, Y is a conjugate, a biomolecule that specifically binds to a target protein.

[0465] Article 77. Conjugate is, formula (V): [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, n is between 1 and 20. m is the average load from 1 to 80. Ab is an antibody or antibody fragment that specifically binds to a target protein. The conjugate as described in Clause 76, wherein Z is a residue portion resulting from the covalent bonding of a chemoselective ligation group to the compatible group of Ab.

[0466] Conjugates described in Clause 76 or 77, where Clause 78.n is 1-6.

[0467] A conjugate as defined in Clause 76 or 77, wherein Clause 79.n is 2 or less.

[0468] The conjugate described in Clause 79, where Clause 80.n is 1.

[0469] A conjugate as described in Clause 76 or 77, wherein Clause 81.n is at least 2.

[0470] The conjugate described in Clause 81, where Clause 82.n is 2.

[0471] The conjugate described in Clause 81, where Clause 83.n is 3.

[0472] The conjugate described in Clause 81, where Clause 84.n is 4.

[0473] A conjugate as defined in any one of the clauses 76-84, which is 1-20, as defined in Clause 85.m.

[0474] A conjugate as described in any one of the clauses 76-84, which is 1-12 of Clause 86.m.

[0475] A conjugate as described in any one of the clauses 76-86, where clause 87.m is at least approximately 2.

[0476] A conjugate as described in any one of the clauses 76-86, where clause 88.m is at least approximately 3.

[0477] A conjugate as described in any one of the clauses 76-86, where clause 89.m is at least approximately 4.

[0478] The conjugate according to any one of the clauses 77-89, wherein clause 90.Z is a residue portion resulting from the covalent bonding of a thiol-reactive chemoselective ligation group to one or more cysteine ​​residues of Ab.

[0479] The conjugate according to any one of the clauses 76 to 89, wherein clause 91.Z is a residue portion resulting from the covalent bonding of an amine-reactive chemoselective ligation group to one or more cysteine ​​residues of Ab.

[0480] Clause 92.X is the part that joins to M6PR, and the expression X: [ka] or a salt thereof, During the ceremony, Each W is independently a hydrophilic head group. each Z 1 However, independently, they are selected from arbitrarily substituted (C1-C3) alkylenes and arbitrarily substituted ethenylenes. each Z 2 However, independently, O, S, NR 21 , and C(R 22 ) Selected from 2, each R 21 However, independently, H is selected from optionally substituted (C1-C6) alkyl groups, and each R 22 However, the conjugate described in any one of the clauses 76 to 91 is independently selected from H, a halogen (e.g., F), and an optionally substituted (C1-C6) alkyl.

[0481] Article 93. The hydrophilic head group W is -OH, -CR 2 R 2 OH, -OP=O(OH)2, -SP=O(OH)2, -NR 3 P=O(OH)2, -OP=O(SH)(OH), -SP=O(SH)(OH), -OP=S(OH)2, -OP=O(N(R 3 )2)(OH), -OP=O(R 3 )(OH), -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 R 4 -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 -SO2NH2, -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 their salts, selected from During the ceremony, R 1 and R 2However, independently, hydrogen, SR 3 , halo, or CN, R 3 and R 4 However, independently, H, C 1-6 Alkyl or substituted C 1-6 It is an alkyl group (e.g., -CF3 or -CH2CF3), A, B, and C are each independently CH or N. A conjugate as described in Clause 92, where D is independently either O or S.

[0482] Clause 94. W is selected from -P=O(OH)2, -SO3H, -CO2H, and -CH(CO2H)2, or salts thereof, as described in Clause 93.

[0483] Clause 95.Z 1 However, -(CH2) j - and j is 1 to 3, a conjugate as described in any one of clauses 92 to 94.

[0484] Clause 96.Z 1 However, a conjugate as described in any one of clauses 92 to 95, where -CH=CH-.

[0485] Clause 97.Z 2 However, a conjugate as defined in any one of clauses 92-96, which is either O or S.

[0486] Clause 98.Z 2 However, -NR 21 -A conjugate as described in any one of the clauses 92 to 96.

[0487] Clause 99.Z 2 However, -C(R 22 )2- A conjugate as described in any one of the clauses 92 to 96.

[0488] Clause 100.Z 1 However, -(CH2) j -, selected from substituted (C1-C3) alkylenes and -CH=CH-, j is 1 to 3, Z 2 However, a conjugate as described in any one of clauses 92-94, selected from O and CH2.

[0489] Clause 101.Z 1 However, these are -(CH2)2-, -CH2-CF2-, or -CH2-CHF-, Z 2 However, the conjugate described in clause 100 is O.

[0490] Clause 102.Z 1 However, these are -(CH2)2-, -CH2-CF2-, or -CH2-CHF-, Z 2 However, CH2 is the conjugate described in Clause 100.

[0491] Clause 103.Z 1 However, -CH=CH-, and Z 2 However, the conjugate described in clause 100 is O.

[0492] Clause 104.Z 1 However, -CH=CH-, and Z 2 However, CH2 is the conjugate described in Clause 100.

[0493] Clause 105.X is a conjugate as defined in any one of Clauses 92-104, selected from the following: [ka]

[0494] Clause 106.X is the part that joins to ASGPR, selected from formulas (III-a) to (III-j): [ka] During the ceremony, R 1 However, -OH, -OC(O)R, and [ka] Selected from, R is C 1-6 It is alkyl, R 2 However, -NHCOCH3, -NHCOCF3, -NHCOCH2CF3, -OH, and [ka] Selected from, R 3 However, a conjugate as described in any one of clauses 76 to 91, selected from -H, -OH, -CH3, -OCH3, and -OCH2CH=CH2.

[0495] The conjugates described in Clause 106, where Clause 107.X is as follows: [ka]

[0496] The conjugates described in Clause 106, where Clause 108.X is as follows: [ka]

[0497] Article 109. Linker L is a linker of formula (IIa), [ka] During the ceremony, Each L 1 ~L 7 However, independently, it is a connecting part, providing a linear or branched linker together between X and Y. a is either 1 or 2, b, c, d, e, f, and g are each independently 0, 1, or 2. Conjugates as described in clauses 76-108, where n is between 1 and 6 (for example, n is between 1 and 5, or 2 and 6, or 1, 2, or 3).

[0498] If clause 110.d is 0, then n is 1, If d is 1, then n is between 1 and 3. If d is 2, then n is 1 to 6, as described in Clause 109.

[0499] Clause 111.-(L 1 ) a - The conjugates described in Clause 109 or 110, which include optionally substituted aryl or heteroaryl conjugate portions.

[0500] Clause 112. Each L 1 However, they became independent, [ka] A conjugate as described in Clause 111, selected from, where v is between 0 and 10 and z is between 0 and 10.

[0501] Clause 113. Each L 2 However, independently, -C 1-6 -Alkilen-, -NHCO-C 1-6 -Alkilen-, -CONH-C 1-6 -Alkylene-, -O(CH2) p -, and -(OCH2CH2) p - is selected from, and p is between 1 and 10. Each L 3 However, they became independent, [ka] , and -(OCH2CH2) q - A conjugate as described in any one of clauses 109 to 112, selected from, where q is between 1 and 10, u is between 0 and 10, and w is between 1 and 10.

[0502] If clause 114.n is 2 or more, then at least one L 4 However, a conjugate described in any one of clauses 109 to 113, which is an existing and branched connecting portion.

[0503] Clause 115. Each L4 However, they became independent, -OCH2CH2-, [ka] , selected from, A conjugate as described in any one of clauses 109 to 114, where each x and y is independently between 1 and 10.

[0504] Article 116. Each L 5 However, it became independent, -NHCO-C 1-6 -Alkilen-, -CONH-C 1-6 -Alkilen-, -C 1-6 -Alkilen-, [ka] , or -(OCH2CH2) r -and, Each L 6 However, it became independent, -NHCO-C 1-6 -Alkilen-, -CONH-C 1-6 -Alkilen-, -C 1-6 -Alkylene-, or -(OCH2CH2)- s -and, Each L 7 However, it became independent, -NHCO-C 1-6 -Alkilen-, -CONH-C 1-6 -Alkilen-, -C 1-6 -Alkilen-, -(OCH2CH2) t -, or -OCH2- A conjugate as described in any one of clauses 109 to 115, where r, s, and t are each independently 1 to 20.

[0505] A conjugate as defined in any one of the clauses 109-116, wherein clause 117.a is 1.

[0506] A conjugate described in any one of the clauses 109-117, where at least one of clauses 118.b, c, e, f, and g is not zero.

[0507] A conjugate as described in any one of Clauses 109-118, wherein at least one of Clauses 119.b or c is not 0, and at least one of e, f, and g is not 0.

[0508] A conjugate as described in any one of the clauses 109-119, wherein clauses 120.a, b, and c are each independently 1 or 2.

[0509] Clause 121. A conjugate described in any one of Clauses 109-120, wherein linker L is selected from any one of the structures in Tables 2-3.

[0510] Clause 122. Conjugates ii) A conjugate derived from the conjugation of any one of the structures in Tables 5-9 and a biomolecule, iii) A conjugate derived from the conjugation of any one of the structures in Tables 5-9 and a polypeptide, or iv) A conjugate as described in Clause 76 or 77, selected from a conjugate derived from a conjugation of any one compound from the structures in Tables 5 to 9 and an antibody or antibody fragment.

[0511] Clause 123. A conjugate as described in any one of Clauses 77-122, wherein the antibody or antibody fragment is an IgG antibody.

[0512] Clause 124. A conjugate as described in any one of Clauses 77-122, wherein the antibody or antibody fragment is a humanized antibody.

[0513] Clause 125. A conjugate according to any one of Clauses 77-124, wherein an antibody or antibody fragment specifically binds to a secreted or soluble protein.

[0514] Clause 126. A conjugate according to any one of Clauses 77 to 124, wherein an antibody or antibody fragment specifically binds to a cell surface receptor.

[0515] Clause 127. A method for internalizing a target protein into a cell comprising a cell surface receptor selected from M6PR and ASGPR, comprising contacting a cell sample comprising cells and a target protein with an effective amount of a compound according to any one of claims 1 to 75 or a conjugate according to any one of claims 76 to 132, wherein the compound or conjugate specifically binds to the target protein and specifically binds to the cell surface receptor to promote cellular uptake of the target protein.

[0516] Clause 128. The method according to Clause 127, wherein the target protein is a membrane-bound protein.

[0517] Clause 129. The method according to Clause 127, wherein the target protein is an extracellular protein.

[0518] Clause 130. The method according to any one of Clauses 127 to 129, wherein the compound or conjugate comprises an antibody or antibody fragment (Ab) that specifically binds to a target protein.

[0519] Clause 131. A method for reducing the level of a target protein in a biological system, comprising contacting the biological system with an effective amount of a compound described in any one of Clauses 1 to 75 or a conjugate described in any one of Clauses 76 to 126, wherein the compound or conjugate specifically binds to the target protein and specifically binds to cell surface receptors of cells in the biological system, thereby promoting the cellular uptake and degradation of the target protein.

[0520] Clause 132. The method according to Clause 131, wherein the biological system comprises cells containing the cell surface receptor M6PR.

[0521] Clause 133. The method according to Clause 131, wherein the biological system includes cells containing the cell surface receptor ASGPR.

[0522] Clause 134. The method described in any one of Clauses 131 to 133, wherein the biological system is a human subject.

[0523] Clause 135. The method described in any one of Clauses 131 to 133, wherein the biological system is an in vitro cell sample.

[0524] Clause 136. The method according to any one of Clauses 131 to 135, wherein the target protein is a membrane-bound protein.

[0525] Clause 137. The method according to any one of Clauses 137 to 135, wherein the target protein is an extracellular protein.

[0526] Clause 138. A method for treating a disease or disorder related to a target protein, comprising administering an effective amount of a compound described in any one of Clauses 1 to 75 or a conjugate described in any one of Clauses 76 to 126 to a subject in need thereof, wherein the compound or conjugate specifically binds to the target protein.

[0527] Clause 139. The method described in Clause 138, wherein the disease or disorder is an inflammatory disease.

[0528] Clause 140. The method described in Clause 138, wherein the disease or disorder is an autoimmune disease.

[0529] Clause 141. The method described in Clause 138, where the disease or disorder is cancer.

[0530] Article 151. Compounds of formula (I): [ka] or a salt thereof, a single stereoisomer, a mixture of stereoisomers, or an isotopic form, During the ceremony, X is the part that binds to the cell surface, L is the linker in equation (IIa) below: [ka] During the ceremony, Each L 1 However, they became independent, [ka] And, Each L 2 However, independently, -C 1-6 -Alkilen-, -NHCO-C 1-6 -Alkilen-, -CONH-C 1-6 -Alkylene-, -(OCH2) p -, or -(OCH2CH2) p -and, Each L 3 However, they became independent, [ka] , or -(OCH2CH2) q -and, Each L 4 However, independently, -OCH2CH2-, [ka] And, Each L 5 However, it became independent, -NHCO-C 1-6 -Alkilen-, -CONH-C 1-6 -Alkilen-, -C 1-6 -Alkilen-, [ka] , or -(OCH2CH2) r -and, Each L 6 However, it became independent, -NHCO-C 1-6 -Alkilen-, -CONH-C 1-6 -Alkilen-, -C 1-6 -Alkylene- or -(OCH2CH2)5- Each L 7 However, it became independent, -NHCO-C 1-6-Alkilen-, -CONH-C 1-6 -Alkilen-, -C 1-6 -Alkilen-, -(OCH2CH2) t -, or -OCH2- p, q, r, s, and t are each independently integers between 1 and 20, a is 1 or 2, b, c, d, e, f, and g are each independently 0, 1, or 2, and u, v, w, x, y, and z are each independently integers between 1 and 10. If n is an integer from 1 to 5 and d is 0, if n is 1 and d is 1, if n is an integer from 1 to 3 and d is 2, if n is an integer from 1 to 5, Y [ka] It is a part selected from the group consisting of, [ka] However, this represents the connection point to L, R is either hydrogen or fluorine. Each R' is independently either a hydrogen atom or a halo. G is selected from -F, -Cl, -Br, -I, -O-mesyl, and -O-tosyl, and J is selected from -Cl, -Br, -I, -F, -OH, -ON-succinimide, -O-(4-nitrophenyl), -O-pentafluorophenyl, -O-tetrafluorophenyl, and -OC(O)-OR J’ Selected from, R J’ However, a compound that is C1-C8 alkyl or aryl.

[0531] Clause 152. The compound according to Clause 151, wherein the cell surface receptor is the cell surface mannose-6-phosphate receptor (M6PR).

[0532] Clause 153. The compound according to Clause 151, wherein the cell surface receptor is a cell surface asialoclycoprotein receptor (ASGPR).

[0533] The compound described in Clause 151, wherein Clause 154.a is 1.

[0534] A compound as described in Clause 151, wherein at least one of b, c, e, f, and g is not 0.

[0535] A compound as described in Clause 151, wherein at least one of Clause 156.b or c is not 0, and at least one of e, f, and g is not 0.

[0536] The compound described in Clause 151, wherein Clause 157.a, b, and c are each independently 1 or 2.

[0537] Clause 158. Each X is independently selected from the group consisting of formulas (IIIa), (IIIb), (IIIc), (IIId), (IIIj), (IIIk), (IIIl), and (IIIm), [ka] In equations (IIIa), (IIIb), (IIIc), or (IIId), R'' is -OH, -CR 1 R 2 OH, -P=O(OH)2, P(=O)R 1 OH, -PH(=O)OH, -(CR 1 R 2 )-P=O(OH)2, -SO2OH, -S(O)OH, -OSO2OH, -COOH, -CONH2, -CONHR 3 ,-CONR 3 R 4 -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 -SO2NH2, -SO2NHR 3 -SO2NR 3 R4 , -SO2NHCOR 3 , -NHCOR 3 ,-NHC(O)NHS(O)2R 3 , -NHSO2R 3 , [ka] Selected from the group consisting of, j is an integer between 1 and 3. R 1 and R 2 However, each is independently hydrogen, halo, or CN. R 3 and R 4 However, each is independent of C 1-6 It is alkyl, A, B, and C are each independently CH or N. Each D is independently either O or S. In equations (IIIj), (IIIk), (IIIl), or (IIIm), R 1 However, -OH, -OC(O)R, or [ka] And R is C 1-6 It is alkyl, R 2 However, -NHCOCH3, -NHCOCF3, -NHCOCH2CF3, -OH, and [ka] Selected from the group consisting of, R 3 The compound described in Clause 151, selected from the group consisting of -H, -OH, -CH3, -OCH3, and -OCH2CH=CH2.

[0538] Clause 159. Each X is independently selected from the group consisting of formulas (IIIa), (IIIb), (IIIc), and (IIId). [ka] During the ceremony, R'' is -OH, -CR 1 R 2 OH, -P=O(OH)2, P(=O)R 1 OH, -PH(=O)OH, -(CR 1 R 2 )-P=O(OH)2, -SO2OH, -S(O)OH, -OSO2OH, -COOH, -CONH2, -CONHR 3 ,-CONR 3 R 4 -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 -SO2NH2, -SO2NHR 3 -SO2NR 3 R 4 , -SO2NHCOR 3 , -NHCOR 3 ,-NHC(O)NHS(O)2R 3 , -NHSO2R 3 , [ka] Selected from the group consisting of, j is an integer between 1 and 3. R 1 and R 2 However, each is independently hydrogen, halo, or CN. R 3 and R 4 However, each is independent of C 1-6 It is alkyl, A, B, and C are each independently CH or N. The compound according to Clause 151, wherein D is independently either O or S.

[0539] Clause 160. Each X is independently selected from the group consisting of formulas (IIIj), (IIIk), (IIIl), and (IIIm), [ka] During the ceremony, R 1 However, -OH, -OC(O)R, or [ka] And R is C 1-6 It is alkyl, R 2 However, -NHCOCH3, -NHCOCF3, -NHCOCH2CF3, -OH, and [ka] Selected from the group consisting of, R 3 The compound described in Clause 151, selected from the group consisting of -H, -OH, -CH3, -OCH3, and -OCH2CH=CH2.

[0540] Article 161. Conjugate of the following formula (IVa): [ka] or a pharmaceutically acceptable salt thereof During the ceremony, X is the part that binds to the cell surface receptor, L is the linker in equation (IIa) below: [ka] During the ceremony, Each L 1 However, they became independent, [ka] And, Each L 2 However, independently, -C 1-6 -Alkilen-, -NHCO-C1-6 -Alkilen-, -CONH-C 1-6 -Alkylene-, -(OCH2) p -, or -(OCH2CH2) p -and, Each L 3 However, they became independent, [ka] , or -(OCH2CH2) q -and, Each L 4 However, independently, -OCH2CH2-, [ka] And, Each L 5 However, it became independent, -NHCO-C 1-6 -Alkilen-, -CONH-C 1-6 -Alkilen-, -C 1-6 -Alkilen-, [ka] , or -(OCH2CH2) r -and, Each L 6 However, it became independent, -NHCO-C 1-6 -Alkilen-, -CONH-C 1-6 -Alkilen-, -C 1-6 -Alkylene- or -(OCH2CH2)5- Each L 7 However, it became independent, -NHCO-C 1-6 -Alkylene-, -CONH-C1-6-Alkylene-, C 1-6 -Alkilen-, -(OCH2CH2) t -, or -OCH2- p, q, r, s, and t are each independently integers between 1 and 20, a is 1 or 2, b, c, d, e, f, and g are each independently 0, 1, or 2, and u, v, w, x, y, and z are each independently integers between 1 and 10. If n is an integer from 1 to 5 and d is 0, if n is 1 and d is 1, if n is an integer from 1 to 3 and d is 2, if n is an integer from 1 to 5, Z [ka] Selected from the group consisting of, [ka] However, this represents the connection point to L, [ka] This represents the connection point to P, X is CH2, NH, O, or S. P stands for polypeptide, or conjugate.

[0541] Clause 162.P comprises an antibody or an antigen-binding fragment of an antibody, as described in Clause 161.

[0542] Article 163. Conjugate the following expression (Va): [ka] or a pharmaceutically acceptable salt thereof During the ceremony, X is the part that binds to the cell surface receptor, L is the linker in equation (IIa) below: [ka] , and in the formula, Each L 1 However, they became independent, [ka] And, Each L 2 However, independently, -C 1-6 -Alkilen-, -NHCO-C 1-6 -Alkilen-, -CONH-C1-6 -Alkylene-, -(OCH2) p -, or -(OCH2CH2) p -and, Each L 3 However, they became independent, [ka] , or -(OCH2CH2) q -and, Each L 4 However, independently, -OCH2CH2-, [ka] And, Each L 5 However, -NHCO-C 1-6 -Alkilen-, -CONH-C 1-6 -Alkilen-, -C 1-6 -Alkilen-, [ka] , or -(OCH2CH2) r -and, Each L 6 However, -NHCO-C 1-6 -Alkilen-, -CONH-C 1-6 -Alkilen-, -C 1-6 -Alkylene-, or -(OCH2CH2)- s -and, Each L 7 However, -NHCO-C 1-6 -Alkilen-, -CONH-C 1-6 -Alkilen-, -C 1-6 -Alkilen-, -(OCH2CH2) t- or -OCH2-, where p, q, r, s, and t are each independently integers from 1 to 20, a is 1 or 2, b, c, d, e, f, and g are each independently 0, 1, or 2, u, v, w, x, y, and z are each independently 1, 2, 3, 4, 5, or 6, n is an integer from 1 to 5, if d is 0, n is 1, if d is 1, n is an integer from 1 to 3, if d is 2, n is an integer from 1 to 5, m is an integer between 1 and 8. Z [ka] Selected from the group consisting of, [ka] However, this represents the connection point to L, [ka] but, [ka] Represents the connection point to, [ka] However, it is an antibody, a conjugate.

[0543] Clause 164. A conjugate as described in any one of Clauses 161 to 163, wherein the cell surface receptor is a cell surface mannose-6-phosphate receptor (M6PR).

[0544] Clause 165. A conjugate as described in any one of Clauses 161 to 163, wherein the cell surface receptor is a cell surface asialoclycoprotein receptor (ASGPR).

[0545] Clause 166. Each X is independently selected from the group consisting of formulas (IIIa), (IIIb), (IIIc), (IIId), (IIIj), (IIIk), (IIIl), and (IIIm), [ka] In equations (IIIa), (IIIb), (IIIc), or (IIId), R'' is -OH, -CR 1 R 2 OH, -P=O(OH)2, P(=O)R 1 OH, -PH(=O)OH, -(CR 1 R 2 )-P=O(OH)2, -SO2OH, -S(O)OH, -OSO2OH, -COOH, -CONH2, -CONHR 3 ,-CONR 3 R 4 -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 -SO2NH2, -SO2NHR 3 -SO2NR 3 R 4 , -SO2NHCOR 3 , -NHCOR 3 ,-NHC(O)NHS(O)2R 3 , -NHSO2R 3 , [ka] Selected from the group consisting of, j is an integer between 1 and 3. R 1 and R 2 However, each is independently hydrogen, halo, or CN. R 3 and R 4 However, each is independent of C 1-6 It is alkyl, A, B, and C are each independently CH or N. Each D is independently either O or S. In equations (IIIj), (IIIk), (IIIl), or (IIIm), R 1 However, -OH, -OC(O)R, or [ka] And R is C 1-6 It is alkyl, R 2 However, -NHCOCH3, -NHCOCF3, -NHCOCH2CF3, -OH, and [ka] Selected from the group consisting of, R 3 A conjugate as described in any one of clauses 161 to 165, selected from the group consisting of -H, -OH, -CH3, -OCH3, and -OCH2CH=CH2.

[0546] Clause 167. Each X is independently selected from the group consisting of formulas (IIIa), (IIIb), (IIIc), and (IIId). [ka] During the ceremony, R'' is -OH, -CR 1 R 2 OH, -P=O(OH)2, P(=O)R 1 OH, -PH(=O)OH, -(CR 1 R 2 )-P=O(OH)2, -SO2OH, -S(O)OH, -OSO2OH, -COOH, -CONH2, -CONHR 3 ,-CONR 3 R 4 -CONH(OH), -CONH(OR 3 ), -CONHSO2R 3 -CONHSO2NR 3 R 4 -CH(COOH)2, -CR 1 R 2 COOH, -SO2R 3 -SOR3 R 4 -SO2NH2, -SO2NHR 3 -SO2NR 3 R 4 , -SO2NHCOR 3 , -NHCOR 3 ,-NHC(O)NHS(O)2R 3 , -NHSO2R 3 , [ka] Selected from the group consisting of, j is an integer between 1 and 3. R 1 and R 2 However, each is independently hydrogen, halo, or CN. R 3 and R 4 However, each is independent of C 1-6 It is alkyl, A, B, and C are each independently CH or N. A conjugate as described in any one of clauses 161-165, where D is independently either O or S.

[0547] Clause 168. Each X is independently selected from the group consisting of formulas (IIIj), (IIIk), (IIIl), and (IIIm), [ka] , in the formula, R 1 However, -OH, -OC(O)R, or [ka] And R is C 1-6 It is alkyl, R 2 However, -NHCOCH3, -NHCOCF3, -NHCOCH2CF3, -OH, and [ka] Selected from the group consisting of, R 3 A conjugate as described in any one of clauses 161 to 165, selected from the group consisting of -H, -OH, -CH3, -OCH3, and -OCH2CH=CH2.

[0548] Clause 169. A pharmaceutical composition comprising a conjugate or pharmaceutically acceptable salt as described in any one of Clauses 161 to 168, and a pharmaceutically acceptable carrier.

[0549] The pharmaceutical composition described in Clause 169, wherein Clause 170.m is an integer between 4 and 8.

[0550] A pharmaceutical composition comprising a conjugate or pharmaceutically acceptable salt described in Clause 170, wherein Clause 171.m is 4.

[0551] Clause 172. A conjugate as described in any one of Clauses 163-168, wherein the antibody is an IgG antibody.

[0552] Clause 173. A conjugate as described in any one of Clauses 163 to 168, wherein the antibody is a humanized antibody.

[0553] Clause 174. A conjugate according to any one of Clauses 163-168, wherein the antibody specifically binds to a secreted or soluble protein.

[0554] Clause 175. A conjugate according to any one of Clauses 163 to 168, wherein the antibody specifically binds to a cell surface receptor.

[0555] Clause 176. A conjugate as described in any one of Clauses 163 to 168, wherein the antibody specifically binds to the programmed death ligand-1 (PD-L1) protein.

[0556] Clause 177. A conjugate as described in any one of Clauses 163-168, wherein the antibody specifically binds to vascular endothelial growth factor (VEGF) protein.

[0557] Clause 178. A conjugate according to any one of Clauses 163 to 168, wherein the antibody specifically binds to fibroblast growth factor receptor 2 (FGFR2) protein or fibroblast growth factor receptor 3 (FGFR3) protein.

[0558] Clause 179. A conjugate as described in any one of Clauses 163-168, wherein the antibody is cetuximab.

[0559] Clause 180. A conjugate as described in any one of Clauses 163-168, wherein the antibody is matsuzumab.

[0560] Clause 181. A conjugate as described in any one of Clauses 163-168, wherein the antibody is atezolizumab.

[0561] Clause 182. A method for treating a disease or disorder by administering an effective amount of a conjugate or pharmaceutically acceptable salt described in any one of Clauses 163 to 168 or a pharmaceutical composition described in Clause 169 to a subject in need of the disease or disorder.

[0562] Clause 183. The method described in Clause 182, wherein the disease or disorder is an inflammatory disease.

[0563] Clause 184. The method described in Clause 182, wherein the disease or disorder is an autoimmune disease.

[0564] Clause 185. The method described in Clause 182, where the disease or disorder is cancer. [Examples]

[0565] The examples in this section are provided for illustrative purposes only, and not as an extension. [Table 11-1] [Table 11-2] [Table 11-3]

[0566] Preparation of compounds The following are illustrative schemes and examples of how the compounds described herein may be prepared and tested. While examples may represent only a limited number of embodiments, it should be understood that the following examples are illustrative and not limiting. All substituents are as previously defined unless otherwise indicated. Reagents and starting materials are readily available to those skilled in the art. Specific synthesis steps for each of the described routes may be combined in different ways or in combination with steps from different schemes to prepare the compounds described herein.

[0567] Mannose-6-phosphate (M6P) ligand Synthesis of Compound A: (2-((2R,3S,4S,5S,6R)-3,4,5-trihydroxy-6-(4-isothiocyanatophenoxy)tetrahydro-2H-pyran-2-yl)ethyl)phosphonic acid (Compound A) [ka]

[0568] (((2R,3S,4S,5R,6R)-2-(4-nitrophenoxy)-6(((trimethylsilyl)oxy)methyl)tetrahydro-2H-pyran-3,4,5-triyl)tris(oxy))tris(trimethylsilane)(A-2)

[0569] A solution of (2R,3S,4S,5S,6R)-2-(hydroxymethyl)-6-(4-nitrophenoxy)tetrahydro-2H-pyran-3,4,5-triol (A-1) (1.0 equivalent, 26.0 g, 86.37 mmol) in DMF (500 mL) was cooled to 0°C. Then, triethylamine (6.4 equivalents, 288 mL, 552.0 mmol) and trimethylsilyl chloride (24.0 equivalents, 70 mL, 2071.0 mmol) were added to the above solution under a nitrogen atmosphere. The resulting mixture was stirred under nitrogen at room temperature for 24 hours. The reaction mixture was partitioned between ethyl acetate and water. The aqueous layer was extracted again with ethyl acetate. The combined organic layers were dried over sodium sulfate, filtered, and purified by silica gel chromatography (0-5% ethyl acetate in hexane) to obtain intermediate A-2 as a colorless oil. Yield: 36.8g (72.3%) 1 H NMR(400MHz,CDCl3)δ8.18(dd,J=12.36,3.16Hz,2H),7.16(dd,J=12.4,3.12Hz,2H),5.37(d ,J=2.36Hz,1H),3.99-3.87(m,3H),3.72-3.69(m,2H),3.50-3.48(m,1H),0.2-0.07(m,36H).

[0570] ((2R,3R,4S,5S,6R)-6-(4-nitrophenoxy)-3,4,5-tris((trimethylsilyl)oxy)tetrahydro-2H-pyran-2-yl)methanol(A-3)

[0571] To a stirred solution of intermediate A-2 (1.0 equivalent, 10.0 g, 16.97 mmol) in a mixture of DCM:methanol (8:2 ratio, 100 mL), ammonium acetate (1.5 equivalents, 1.96 g, 25.46 mmol) was added under nitrogen at room temperature. The resulting mixture was stirred under nitrogen at room temperature for 16 hours. The reaction mixture was partitioned between ethyl acetate and water. The aqueous layer was extracted again with ethyl acetate. The combined organic layers were dried over sodium sulfate, filtered, concentrated under vacuum, and purified by silica gel chromatography (20-30% ethyl acetate in hexane) to obtain intermediate A-3 as a white solid. Yield: 7.0 g (80%); LC-MS m / z 516.13 [M-1] - .

[0572] (2S,3R,4S,5S,6R)-6-(4-nitrophenoxy)-3,4,5-tris((trimethylsilyl)oxy)tetrahydro-2H-pyran-2-carbaldehyde(A-4)

[0573] To a stirred solution of oxalyl chloride (1.1 equivalents, 0.5 mL, 5.31 mmol) in DCM (5 mL), a solution of DMSO (2.2 equivalents, 0.76 mL, 10.62 mmol) in DCM (5 mL) was added over 5 minutes at -78°C. After stirring at -78°C for 20 minutes, a solution of intermediate A-3 (1.0 equivalent, 2.5 g, 4.83 mmol) in DCM (10 mL) was added to the mixture. The reaction mixture was further stirred at -78°C for 60 minutes, followed by the addition of triethylamine (5.0 equivalents, 3.4 mL, 24.15 mmol). The resulting mixture was allowed to reach room temperature over 1 hour. The turbid mixture was diluted with DCM, washed with water, and then washed with brine. The organic layer was dried over sodium sulfate, filtered, and concentrated under high vacuum to obtain intermediate A-4 (2.2 g, crude) as a light brown gel, which was used in the next step without further purification.

[0574] Diethyl((E)-2-((2R,3R,4S,5S,6R)-6-(4-nitrophenoxy)-3,4,5-tris((trimethylsilyl)oxy)tetrahydro-2H-pyran-2-yl)vinyl)phosphonate(A-5)

[0575] A stirred suspension of tetraethylmethylenebis(phosphonate) (1.5 equivalents, 1.85 g, 6.40 mmol) in dry THF (20 mL) was cooled to -78°C, and n-BuLi (1.25 equivalents, 2.6 ml, 5.33 mmol) in 2.0 M hexane was added. The resulting mixture was stirred at -78°C for 1 hour, and then intermediate A-4 (1.0 equivalent, 2.2 g, 4.27 mmol) in dry THF (10 mL) was added at -78°C. The bath was removed, the reaction mixture was allowed to come to room temperature, and stirred for 12 hours. Saturated aqueous NH4Cl solution was added, and the mixture was extracted with ethyl acetate. The ethyl acetate layer was washed with water, followed by washing with saturated brine. The organic layer was dried over sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel chromatography (30-40% ethyl acetate in hexane) to obtain intermediate A-5 as a colorless gel. Yield (1.3g, 48%); LC-MS m / z 650.57[M+1] + .

[0576] Diethyl((E)-2-((2R,3S,4S,5S,6R)-3,4,5-trihydroxy-6-(4-nitrophenoxy)tetrahydro-2H-pyran-2-yl)vinyl)phosphonate(A-6)

[0577] To a stirred solution of intermediate A-5 (1.0 equivalent, 1.3 g, 1.54 mmol) in methanol (15 mL), Dowex 50WX8 hydrogen type was added at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at room temperature under nitrogen for 2 hours. The reaction mixture was filtered, washed with methanol, and the filtrate was concentrated under vacuum to obtain diethyl((E)-2-((2R,3S,4S,5S,6R)-3,4,5-trihydroxy-6-(4-nitrophenoxy)tetrahydro-2H-pyran-2-yl)vinyl)phosphonate (6) as a white solid. Yield: 0.78 g (90%); LC-MS m / z 434.17 [M+1] + .

[0578] (2R,3R,4S,5S,6R)-2-((E)-2-(diethoxyphosphoryl)vinyl)-6-(4-nitrophenoxy)tetrahydro-2H-pyran-3,4,5-triyltriacetate(A-7)

[0579] To a stirred solution of intermediate A-6 (1.00 equivalent, 0.78 g, 1.80 mmol) in pyridine (10 mL), acetic anhydride (10.0 equivalent, 1.8 mL, 18.0 mmol) was added dropwise at 0°C under nitrogen. The cooling bath was removed, and the resulting mixture was stirred under nitrogen at room temperature for 16 hours. Pyridine was removed under high vacuum, and the residue was partitioned between ethyl acetate and aqueous 1N HCl. The aqueous layer was extracted again with ethyl acetate. The combined organic layers were dried over sodium sulfate, filtered, concentrated, and purified by silica gel chromatography (2.5% methanol in dichloromethane) to obtain intermediate A-7 as a white solid. Yield: 1.0 g (100%); LC-MS m / z 560.17 [M+1] + .

[0580] (2R,3S,4S,5R,6R)-2-(4-aminophenoxy)-6-(2-(diethoxyphosphoryl)ethyl)tetrahydro-2H-pyran-3,4,5-triyltriacetate(A-8)

[0581] To a stirred solution of intermediate A-7 (1.0 equivalent, 1.0 g, 1.78 mmol) in methanol (15 mL), 10% palladium carbon (0.200 g) was added under nitrogen at room temperature. The resulting mixture was stirred at room temperature for 16 hours under hydrogen gas pressure (100 psi). The reaction mixture was filtered through a Celite bed, washed with methanol, and the filtrate was concentrated under vacuum to obtain intermediate A-8 as a brown, viscous gel. Yield: 0.700 g (73.6%); LC-MS m / z 532.21 [M+1] + .

[0582] (2-((2R,3R,4S,5S,6R)-3,4,5-triacetoxy-6-(4-aminophenoxy)tetrahydro-2H-pyran-2-yl)ethyl)phosphonic acid (A-9)

[0583] To a stirred solution of intermediate A-8 (1.00 equivalent, 2.0 g, 5.73 mmol) in acetonitrile (15 mL), bromotrimethylsilane (5.0 equivalent, 3.8 mL, 28.65 mmol) was added dropwise at 0°C under nitrogen. The cooling bath was removed, and the resulting mixture was stirred under nitrogen at room temperature for 16 hours. Volatile substances were removed using a rotary evaporator, and the residue was dried under high vacuum. The crude residue was pulverized with diethyl ether and dried under high vacuum to obtain intermediate A-9 as a brown solid. Yield: 2.2 g, crude. LC-MS m / z 476.0 [M+1] + .

[0584] (2-((2R,3S,4S,5S,6R)-6-(4-aminophenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)ethyl)phosphonic acid (A-10)

[0585] To a stirred solution of intermediate A-9 (1.0 equivalent, 2.0 g, 4.21 mmol) in a methanol:water mixture (8:2, 15 mL), triethylamine (5.0 equivalent, 2.93 mL, 21.05 mmol) was added dropwise at 0°C under nitrogen. The cooling bath was removed, and the resulting mixture was stirred at room temperature for 16 hours. Methanol was removed using a rotary evaporator, and the residue was dried under high vacuum. The residue was taken into water and purified by preparative HPLC (2-10% acetonitrile in water containing 5 mM ammonium acetate). The fractions containing the desired product were combined and freeze-dried to obtain intermediate A-10 as a brown solid. Yield: 0.350 g (25%); LC-MS m / z 348.0 [MH] - .

[0586] (2-((2R,3S,4S,5S,6R)-3,4,5-trihydroxy-6-(4-isothiocyanatophenoxy)tetrahydro-2H-pyran-2-yl)ethyl)phosphonic acid (compound A)

[0587] To a stirred solution of intermediate A-10 (1.0 equivalent, 1.75 g, 5.01 mmol) in a mixture of ethanol:water (7:3) (20 ml), thiophosgene (5.00 equivalent, 1.92 mL, 25.05 mmol) was added dropwise at 0°C under nitrogen. The cooling bath was removed, and the resulting mixture was stirred under nitrogen at room temperature for 3 hours. Volatile substances were removed using a rotary evaporator, and the residue was dried under high vacuum. The residue was taken into water and purified by preparative HPLC (20-40% acetonitrile in water containing 5.0 mmol of ammonium acetate). The fractions containing the desired product were combined and freeze-dried to obtain compound A as a white solid. Yield: 0.135 g (6.8%) LC-MS m / z 392.08 [M+1] + ; 1 H NMR(400MHz,D2O)δ7.32(d,J=8.92Hz,2H),7.12(d,J=8.96Hz,2H),5.57(s,1H),4.13(s,1H),3.96(d d,J=9.16,3.44Hz,1H),3.59-3.48(m,2H),2.03-1.88(m,1H),1.68-1.54(m,2H),1.27-1.15(m,1H).

[0588] Example 1: Synthesis of Compound I-1 [ka]

[0589] A solution of 3,3'-(ethane-1,2-diyrbis(oxy))dipropionic acid (1A) (1.0 equivalent, 0.200 g, 0.96 mmol) and 2,3,5,6-tetrafluorophenol (2.0 equivalent, 0.315 g, 1.9 mmol) in ethyl acetate (4 mL) was cooled to 0°C. N,N'-diisopropylcarbodiimide (3.0 equivalent, 0.44 mL, 2.8 mmol) was added, and the reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was filtered through a Celite bed, and the Celite bed was washed with ethyl acetate. The filtrate was concentrated to obtain the crude product, which was purified by column chromatography using silica gel (100-200 mesh) and 0-10% ethyl acetate in hexane to obtain compound 1B as a colorless viscous liquid. Yield: 0.370 g, 76.1%; LC-MS m / z 500.96 [M-1] - .

[0590] Intermediate A-10 (1.0 equivalent, 0.040 g, 0.11 mmol) was dissolved in dimethyl sulfoxide (1 mL), and triethylamine (10.0 equivalent, 0.15 mL, 1.1 mmol) was added. In a separate vial, compound 1B (5.0 equivalent, 0.276 g, 0.55 mmol) was dissolved in dimethyl sulfoxide (1 mL), and the previous mixture was added dropwise to this mixture (over 30 minutes). The reaction mixture was stirred at room temperature for 5 minutes. After completion, the reaction mixture was diluted with acetonitrile and purified by preparative HPLC (25-45% acetonitrile in water containing 0.1% TFA). The fractions containing the desired product were combined and lyophilized to dryness to obtain compound I-1 as an off-white solid. Yield: 0.002 g, 2.5%; LC-MS m / z 686.25 [M+1] + ; 1H NMR(400MHz,D2O)δ7.35(d,J=8.88Hz,2H),7.29-7.23(m,1H),7.07(d,J=8.96Hz ,2H),5.50(s,1H),4.13(bs,1H),3.98-3.95(m,1H),3.91(t,J=5.64Hz,2H),3.8 6(t,J=5.72Hz,2H),3.72(s,4H),3.58(d,J=7.32Hz,2H),2.96(t,J=5.76Hz,2H) ,2.66(t,J=5.8Hz,2H),2.03-2.00(m,1H),1.74-1.63(m,2H),1.32-1.26(m,1H).

[0591] Example 2: Synthesis of Compound I-2 [ka]

[0592] To a stirred solution of 1-(9H-fluoren-9-yl)-3-oxo-2,7,10-trioxa-4-azatridecane-13-euic acid (2A) (2.0 g, 5.00 mmol) in acetonitrile (16 mL), piperidine (4 mL) was added, and the reaction mixture was stirred for 1 hour. The progress of the reaction was monitored by TLC. After the completion of the reaction, the reaction mixture was concentrated to obtain the crude product. The crude product was washed with hexane and dried to obtain compound 2B as an off-white semi-solid. Yield: 0.85 g, 96%; LC-MS m / z 178.06 [M+1] + .

[0593] To a stirred solution of 2,5-dioxopyrrolidine-1-yl-6-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)hexanoate (2C) (1.0 g, 3.24 mmol) and compound 2B (0.86 g, 4.87 mmol) in N,N-dimethylformamide (20 mL), N,N-diisopropylethylamine (1.46 mL, 8.11 mmol) was added, and the reaction mixture was stirred for 3 hours. The progress of the reaction was monitored by LC-MS. After the completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain the crude product. The crude product was purified by preparative HPLC (XBS column using 30% ACN in 70% 5 mM ammonium acetate) to obtain compound 2D as brown oil. Yield: 0.6 g, 43%; LC-MS m / z 371.22 [M+1] + .

[0594] To a stirred solution of compound 2D (0.35 g, 0.945 mmol) and pentafluorophenol (0.17 g, 0.945 mmol) in ethyl acetate (10 mL), N,N'-diisopropylcarbodiimide (0.13 g, 1.04 mmol) was added, and the reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by TLC and LC-MS. After the reaction was complete, the reaction mixture was filtered through a filter cartridge, washed with a small amount of ethyl acetate (2 mL), and concentrated under reduced pressure under an inert atmosphere to obtain crude compound 2E, which was used in the next step without further purification. Yield: 0.2 g, (crude); LC-MS m / z 537.19 [M+1] + .

[0595] To a stirred solution of intermediate A-10 (0.05 g, 0.14 mmol), powder molecular sieve, and compound 2E (0.11 g, 0.21 mmol) in dimethyl sulfoxide (2 mL), triethylamine (0.04 g, 0.42 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by LC-MS. The reaction mixture was purified by preparative HPLC (XB-C-18 column using 40% ACN in 60% 5 mM ammonium acetate). The fractions containing the desired product were combined and lyophilized to dryness to obtain compound I-2 as a white solid. Yield: 0.03 g, 31%; LC-MS m / z 702.31 [M+1] + . 1 H NMR(400MHz,D2O)δ7.37(d,J=8.8Hz,2H),7.11(d,J=8.9Hz,2H),6.77(s,2H),5.52(d,J=1.4 8Hz,1H),5.20(bs,1H),4.14-4.13(m,1H),3.98-3.95(m,1H),3.85(t,J=5.88Hz,2H),3.72- 3.66(m,6H),3.60-3.55(m,4H),3.42(t,J=6.96Hz,2H),3.29(t,J=5.28Hz,2H),2.66(t,J=5 .84Hz,2H),2.10(t,J=7.32Hz,2H),2.04-1.95(m,1H),1.69-1.57(m,2H),1.54-1.45(m,4H).

[0596] Example 3: Synthesis of Compound I-3 [ka]

[0597] Piperidine (1 mL) was added at room temperature to a stirred solution of 1-(9H-fluoren-9-yl)-3-oxo-2,7,10,13,16,19,22,25,28,31,34,37,40-tridecaoxa-4-azatritetracontane-43-euic acid (3A) (1.0 g, 1.19 mmol) in acetonitrile (9 mL), and the reaction was maintained for 1 hour. The progress of the reaction was monitored by TLC. After completion, the reaction mixture was concentrated to obtain the crude residue. The residue was washed with hexane (10 mL x 4) and dried under vacuum to obtain compound 3B (0.700 g, 95%) as an off-white solid. 1 ¹H NMR (400 MHz, dimethyl sulfoxide-d6): δ 3.60-3.45 (m, 48H), 2.75 (t, J=5.7Hz, 2H), 2.28 (t, J=6.7Hz, 2H).

[0598] At room temperature, a stirring solution of compounds 3B (0.600 g, 0.971 mmol) and 2C (0.449 g, 1.46 mmol) in N,N-dimethylformamide (10 mL) was mixed with N,N-diisopropylethylamine (0.448 mL, 2.43 mmol), and the reaction mixture was stirred for 3 hours. After the reaction was complete, the reaction mixture was concentrated under reduced pressure to obtain a concentrated residue. The residue was purified by preparative HPLC using an XB-C18 (19 x 250 mm) 10 μL column containing 20-45% acetonitrile as the eluent in water containing 5 mM ammonium acetate buffer. The desired fractions were combined and lyophilized to obtain compound 3C as a pale yellow oil. LC-MS m / z 809.5 [M-1] - Yield: 0.433g, 55%.

[0599] To a stirred solution of compound 3C (0.15 g, 0.185 mmol) and pentafluorophenol (0.040 g, 0.222 mmol) in ethyl acetate (5 mL), N,N'-diisopropylcarbodiimide (0.028 g, 0.222 mmol) was added at 0°C, and the reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by TLC and LC-MS. After the completion of the reaction, the solid observed by diisopropylurea in the reaction mixture was filtered through a filter cartridge, washed with a small amount of ethyl acetate (2 mL), and concentrated under vacuum to obtain crude compound 3D, which was not further purified for the next step. LC-MS m / z 994.5 [M + H2O] + Yield: 0.120g, 66%.

[0600] Intermediate A-10 (0.032 g, 0.091 mmol) in dimethyl sulfoxide (0.5 mL) was added dropwise to a stirred solution of compound 3D (0.099 g, 0.101 mmol) in dimethyl sulfoxide (0.5 mL) at room temperature, and the mixture was stirred for 5 minutes. Triethiamine (0.013 g, 0.137 mmol) was added to the reaction mixture, and the reaction was maintained at room temperature for 16 hours. Subsequently, preparative HPLC was performed using a Sunfire C18 (19 × 250 mm) 10 μF column containing 0.1% TFA as the eluent in water and 40-60% acetonitrile. The fractions containing the desired product were combined and lyophilized to dryness to obtain the desired compound I-3 (0.011 g, yield 10%) as a concentrated syrup. LC-MS m / z 1142.6 [M+1] + . 1 H NMR(400MHz,D2O)δ7.27(d,J=9.0Hz,2H),7.16(d,J=9.0Hz,2H)6.83(s,2H),5.56( s,1H),4.20-4.15(m,1H),4.05-3.98(m,1H),3.88(t,J=6.0Hz,2H),3.75-3.58(m,4 9H),3.49(t,J=6.8Hz,2H),3.37(t,J=5.6Hz,2H),2.69(t,J=6.0Hz,2H),2.23(t,J =7.2Hz,2H),2.10-1.98(m,1H),1.75-1.55(m,6H),5.56(s,1H),1.38-1.20(m,2H).

[0601] Example 4: Synthesis of Compound I-4 [ka]

[0602] A solution of 6-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)hexanoic acid (4A) (1.0 equivalent, 2.5 g, 11.8 mmol) and 2,3,4,5,6-pentafluorophenol (1.0 equivalent, 2.17 g, 11.8 mmol) in ethyl acetate (50 mL) was cooled to 0°C. N,N'-diisopropylcarbodiimide (1.1 equivalent, 2.0 mL, 12.9 mmol) was added, and the reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was filtered through a Celite bed and washed with ethyl acetate. The filtrate was concentrated to obtain the crude product, which was purified by column chromatography using silica gel (100-200 mesh) and 0-25% ethyl acetate in hexane to obtain compound 4B as a white solid. Yield: 3.50 g, 79.5%; LC-MS m / z 377.99 [M+1] + .

[0603] Intermediate A-10 (1.0 equivalent, 0.050 g, 0.14 mmol) was dissolved in dimethyl sulfoxide (1 mL), and triethylamine (3.0 equivalent, 0.06 mL, 0.42 mmol) and compound 4B (2.0 equivalent, 0.105 g, 0.28 mmol) were added. The reaction mixture was stirred at room temperature for 16 hours. After completion, the reaction mixture was diluted with acetonitrile and purified by preparative HPLC (8-15% acetonitrile in water containing 5 mM ammonium acetate). The fractions containing the desired product were combined and freeze-dried to obtain compound I-4 as an off-white solid. Yield: 0.006 g, 8.0%; LC-MS m / z 543.27 [M+1] + ; 1H NMR(400MHz,D2O)δ7.35(d,J=8.96Hz,2H),7.16(d,J=9.0Hz,2H),6.77(s,2H),5.58(d,J=1.64Hz,1H),4.17-4.16(m,1H),4.02-3.95 (m,1H),3.63-3.57(m,2H),3.52(t,J=6.84Hz,2H),2.39(t,J=7.24Hz,2H),2.06-1.98(m,1H),1.74-1.58(m,6H),1.37-1.22(m,3H).

[0604] Example 5: Synthesis of Compound I-5 [ka]

[0605] In dimethyl sulfoxide (1.0 mL), a molecular sieve (powder, catalyst support, sodium Y zeolite, Aldrich catalog number 334448) was added, followed by intermediate A-10 (1.0 equivalent, 0.060 g, 0.172 mmol), triethylamine (3.0 equivalent, 0.074 mL, 0.515 mmol), and 2,5-dioxopyrrolidine-1-yl 3-(2-(2-(prop-2-in-1-yloxy)ethoxy)ethoxy)propanoate (5A) (1.0 equivalent, 0.053 g, 0.172 mmol). The reaction mixture was stirred at room temperature for 3 hours. After completion, the reaction mixture was diluted with acetonitrile and purified by preparative HPLC (14-33% acetonitrile in water containing 0.1% TFA). The fractions containing the desired product were combined and freeze-dried to obtain compound 5B as an off-white, sticky solid. Yield: 0.018 g, 17.93%; LC-MS m / z 548.32 [M+1] + .

[0606] A solution of compound 5B (1.0 equivalent, 0.018 g, 0.032 mmol) and perfluorophenyl 3-(2-(2-azidoethoxy)ethoxy)propanoate (5C) (1.2 equivalents, 0.014 g, 0.039 mmol) in dimethyl sulfoxide (0.6 mL) was stirred at room temperature for 5 minutes. Then, tetrakis(acetonitrile)copper(I)hexafluorophosphate (2.8 equivalents, 0.034 g, 0.092 mmol) was added, and the reaction mixture was stirred at room temperature for 1 hour. After completion, the reaction mixture was diluted with acetonitrile and purified by preparative HPLC (40-60% acetonitrile in water containing 0.1% TFA). The fractions containing the desired product were combined and freeze-dried to dryness to obtain compound I-5 as a white solid. Yield: 0.015g, 48.67%; LC-MS m / z 917.37[M+1] + ; 1 H NMR(400MHz,D2O)δ7.97(s,1H),7.36(d,J=9.2Hz,2H),7.08(d,J=9.2Hz,2H),5.51(s,1H),4.59-4.55(m,2H),4.15-4.14(m,1H),3.97-3.92 (m,3H),3.87-3.81(m,4H),3.70-3.57(m,14H),2.97(t,J=6.0Hz,2H),2.66(t,J=6.0Hz,2H),2.00(bs,1H),1.71-1.64(m,2H),1.33(bs,1H).

[0607] Example 6: Synthesis of Compound I-6 [ka]

[0608] To a stirred solution of intermediate A-10 (0.02 g, 0.057 mmol) in dimethyl sulfoxide (2 mL), filtered by powder molecular sieving, and 2,5-dioxopyrrolidine-1-yl-1-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)-3,6,9,12,15,18,21,24,27,30,33,36-dodecaoxanonateacontan-39-oate (6A) 0.07 g, 0.085 mmol), triethylamine (0.018 g, 0.172 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by LC-MS. The reaction mixture was purified by preparative HPLC (Xselect-phenylhexyl using 0.1% TFA in 30% ACN and 70% H2O). The fractions containing the desired product were combined and freeze-dried to obtain compound I-6 as a white solid. Yield: 0.0065 g, 11%; LC-MS m / z 1029.58 [M+1] + . 1 H NMR(400MHz,D2O)δ7.42(d,J=8.8Hz,2H),7.16(d,J=9.2Hz,2H),6.86(s,2H),5.56(s,1H),4.16(d,J=1.6Hz 1H),4.00(t,J=9.6Hz 1H),3.87(t,J=5.88Hz,2H),3.71-3.61(m,50H),2.69(t,J=11.6Hz,2H),2.15-1.95(m,1H),1.75-1.61(m,2H),1.43-1.25(m,1H).

[0609] Example 7: Synthesis of Compound I-7 [ka]

[0610] A solution of hexy-5-en-1-amine (7A) (1.20 equivalents, 3.9 mg, 0.0405 mmol) in NMP (0.15 mL) was added to compound A (1.00 equivalent, 13.2 mg, 0.0337 mmol) in a drum vial equipped with a stirring rod. The resulting mixture was capped and stirred at room temperature for 30 minutes (the solid was slowly dissolved to obtain a clear yellow solution). A solution of azide-PEG4-pentafluorophenol ester (7B) (1.50 equivalents, 23.1 mg, 0.0506 mmol) in NMP (0.20 mL) was added, followed by tetrakis(acetonitrile)copper(I) hexafluorophosphate (3.00 equivalents, 37.7 mg, 0.101 mmol). The resulting clear dark yellow solution was capped and stirred at room temperature for 30 minutes. The reaction mixture was diluted with a mixture of NMP, ethanol, and acetic acid, filtered, and purified by preparative HPLC (15-60% acetonitrile in water containing 0.1% TFA). The fractions containing the desired product were combined and freeze-dried to obtain compound I-7 as a white solid. Yield: 11.1 mg, 35%; LC-MS m / z 946.5[M+1]+; 1 H NMR (300MHz, DMSO-d6 containing D2O) δ7.80(s,1H),7.25(d,J=8.4Hz,2H),6.98(d,J=8.4Hz,2H),5.32(s,1H),4.44(s,2H),3.86-3.6 8(m,5H),3.67-3.23(m,17H),3.05-2.91(m,2H),2.67-2.56(m,2H),2.00-1.81(m,1H),1.69-1.41(m,6H),1.30-1.07(m,1H).

[0611] Example 8: Synthesis of Compound I-8 [ka]

[0612] DBU (0.05 equivalents, 0.025 mL, 0.168 mmol) was added at 0°C under nitrogen to a stirred solution of (2R,3R,4S,5S,6S)-2-(2-(diethoxyphosphoryl)ethyl)-6-hydroxytetrahydro-2H-pyran-3,4,5-triyltriacetate (8A) (1.00 equivalents, 1.48 g, 3.36 mmol) and trichloroacetonitrile (10.0 equivalents, 3.4 mL, 33.6 mmol) in DCM (30 mL). The resulting mixture was stirred at 0°C under nitrogen. Further, DBU (0.0500 equivalents, 0.025 mL, 0.168 mmol) was added, and the cooling bath was removed. The resulting mixture was stirred at room temperature for 45 minutes. Most of the solvent was removed using a rotary evaporator. The residue was loaded onto a silica gel loading column pre-equilibrated with 0.1% triethylamine in dichloromethane and purified by silica gel chromatography (column pre-equilibrated with 0.1% triethylamine in 30% ethyl acetate / hexane) (30-100% ethyl acetate in hexane). The fractions containing the desired product were combined and concentrated using a rotary evaporator. The residue was removed twice from dry dichloromethane, dried under high vacuum for 30 minutes, and then stored at -80°C under nitrogen to obtain compound 8B as a colorless semi-solid. Yield: 1.26 g, 64%; 1 ¹H NMR (300MHz, chloroform-d): δ 8.74 (s, 1H), 6.21 (s, 1H), 5.45 (s, 1H), 5.34 (t, J=11.2Hz, 1H), 5.20 (t, J=10.0Hz, 1H), 4.16-4.00 (m, 4H), 4.00-3.88 (m, 1H), 2.18 (s, 3H), 2.07 (s, 3H), 2.00 (s, 3H), 1.95-1.64 (m, 4H), 1.31 (t, J=7.3Hz, 6H).

[0613] Compound 8B (1.00 equivalent, 1.25 g, 2.14 mmol) was dissolved in dry DCM (10 mL) under nitrogen with stirring. Buta-3-en-1-ol (2.00 equivalent, 0.32 mL, 4.28 mmol) was added, and the resulting mixture was cooled to -78°C under nitrogen with stirring. A solution of boron trifluoride diethyl ether (0.500 equivalent, 0.13 mL, 1.07 mmol) in dichloromethane (5 mL) was slowly added. The -78°C cooling bath was removed, and the reaction mixture was slowly warmed under nitrogen for 50 minutes. The reaction mixture was cooled in a water / ice bath, stirred at 0°C under nitrogen for a further 30 minutes, and then work-up was performed. The reaction mixture was partitioned between dichloromethane and saturated sodium bicarbonate aqueous solution. The aqueous layer was extracted again with dichloromethane. The combined organic matter was dried over sodium sulfate, filtered, and purified by silica gel chromatography (20-100% ethyl acetate in dichloromethane) to obtain compound 8C as a colorless viscous oil. Yield: 408 mg, 39%; LC-MS m / z 493.4[M+1]+; 1 H NMR(300MHz,chloroform-d)δ5.35-5.19(m,2H),5.09(t,J=9.9Hz,1H),4.79(s,1H),4.21-3.98(m,4H),3.91-3.68(m,2H), 3.64-3.50(m,1H),2.55-2.44(m,2H),2.15(s,3H),2.05(s,3H),1.98(s,3H),2.07-1.62(m,5H),1.32(t,J=7.2Hz,6H).

[0614] Bromotrimethylsilane (5.00 equivalents, 0.47 mL, 3.57 mmol) was slowly added at 0°C under nitrogen to a stirred solution of compound 8C (1.00 equivalent, 352 mg, 0.715 mmol) in MeCN (7 mL). The cooling bath was removed, and the resulting mixture was stirred under nitrogen at room temperature for 3.5 hours. Volatile substances were removed using a rotary evaporator, and the residue was dried briefly under high vacuum. The residue was dissolved in methanol (7 mL) while stirring under nitrogen, and sodium methoxide (25% by weight in methanol) (2.50 equivalents, 0.41 mL, 1.79 mmol) was added. The resulting mixture was stirred under nitrogen at room temperature for 1 hour. Acetic acid (3.00 equivalents, 0.12 mL, 2.14 mmol) was added, and then volatile substances were removed using a rotary evaporator. The residue was taken into water and purified by preparative HPLC (0-15% acetonitrile in water containing 0.1% TFA). Most of the solvent was removed using a rotary evaporator at 30°C, and the remainder was freeze-dried to obtain compound 8D as a white solid. Yield: 208 mg, 94%; LC-MS m / z 311.3[M+1]+; 1 H NMR (300MHz, deuterium oxide) δ4.88-4.80(m,1H),3.93(s,1H),3.84-3.70(m,2H),3.70-3 .56(m,2H),3.48(t,J=9.7Hz,1H),2.57-2.44(m,2H),2.37(s,1H),2.15-1.61(m,4H).

[0615] Compound 8D (1.00 equivalent, 10.0 mg, 0.032 mmol) and azide-PEG4-pentafluorophenol ester 7B (1.20 equivalent, 17.7 mg, 0.039 mmol) were dissolved in NMP (0.3 mL) with stirring. After 2 minutes, tetrakis(acetonitrile)copper(I)hexafluorophosphate (2.80 equivalent, 33.6 mg, 0.090 mmol) was added. The resulting pale yellow solution was covered and stirred at room temperature for 30 minutes (it slowly turned greener). The reaction mixture was diluted with a mixture of NMP, ethanol, and acetic acid, filtered, and purified by preparative HPLC (15-65% acetonitrile in water containing 0.1% TFA). The fractions containing the desired product were combined and lyophilized to dryness to obtain compound I-8 as a white solid. Yield: 12.3 mg, 50%; LC-MS m / z 768.5[M+1]+; 1 H NMR(300MHz,DMSO-d6)δ7.81(s,1H),4.59(s,1H),4.44(bs,2H),3.60-3.30(m,1 7H), 3.27-2.76 (m, 9H), 2.01-1.84 (m, 1H), 1.77-1.58 (m, 1H), 1.56-1.32 (m, 2H).

[0616] Example 9: Synthesis of Compound I-9 [ka]

[0617] Compound 8D (1.00 equivalent, 9.8 mg, 0.0316 mmol) and azide-PEG8-pentafluorophenol ester (9A) (1.20 equivalent, 24.0 mg, 0.0379 mmol) were dissolved in NMP (0.3000 mL) with stirring. After 2 minutes, tetrakis(acetonitrile)copper(I)hexafluorophosphate (2.80 equivalent, 33.0 mg, 0.0884 mmol) was added. The resulting pale yellow solution was covered and stirred at room temperature for 30 minutes (it slowly turned greener). The reaction mixture was diluted with a mixture of NMP, ethanol, and acetic acid, filtered, and purified by preparative HPLC (15-65% acetonitrile in water containing 0.1% TFA). The fractions containing the desired product were combined and lyophilized to dryness to obtain compound I-9 as a white solid. Yield: 18.9 mg, 63%; LC-MS m / z 944.6[M+1]+; 1 1H NMR (300MHz, DMSO-d6 containing D2O): δ 7.81 (s,1H), 4.59 (s,1H), 4.44 (s,2H), 3.86-3.29 (m,34H), 3.29-2.69 (m,8H), 2.01-1.80 (m,1H), 1.80-1.57 (m,1H), 1.56-1.30 (m,2H).

[0618] Example 10: Synthesis of Compound I-10 [ka]

[0619] A solution of azido-PEG3-amine (10B) (1.30 equivalents, 14.3 mg, 0.0654 mmol) in NMP (0.3000 mL) was added to 2,5-dioxopyrrolidine-1-yl3-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)propanoate (10A) (1.30 equivalents, 17.4 mg, 0.0654 mmol) in a 1-drum vial equipped with a stirring bar. The resulting colorless, clear solution was capped and stirred at room temperature for 30 minutes, then added to compound 8D (1.00 equivalents, 15.6 mg, 0.0503 mmol) in a 1-drum vial equipped with a stirring bar. After 2 minutes, tetrakis(acetonitrile)copper(I)hexafluorophosphate (2.80 equivalents, 52.5 mg, 0.141 mmol) was added. The resulting pale yellow solution was capped and stirred at room temperature for 30 minutes. The reaction mixture was diluted with a mixture of NMP, ethanol, and acetic acid, filtered, and purified by preparative HPLC (5-40% acetonitrile in water containing 0.1% TFA). The fractions containing the desired product were combined and freeze-dried to obtain compound I-10 as a white solid. Yield: 17.7 mg, 52%; LC-MS m / z 680.5[M+1]+; 1 H NMR (300MHz, DMSO-d6 containing D2O) δ7.81(s,1H),6.92(s,2H),4.59(s,1H),4.44(s,2H),3.63-3.26(m,1 5H), 3.26-2.70 (m, 9H), 2.36-2.21 (m, 2H), 2.05-1.83 (m, 1H), 1.79-1.60 (m, 1H), 1.54-1.30 (m, 2H).

[0620] Example 11: Synthesis of Compound I-11 [ka]

[0621] Compound 8D (1.00 equivalent, 13.4 mg, 0.0432 mmol) and azide-PEG1-pentafluorophenol ester (11A) (1.20 equivalent, 16.9 mg, 0.0518 mmol) were dissolved in NMP (0.3000 mL) with stirring. After 2 minutes, tetrakis(acetonitrile)copper(I)hexafluorophosphate (2.80 equivalent, 45.1 mg, 0.121 mmol) was added. The resulting pale yellow solution was covered and stirred at room temperature for 30 minutes. The reaction mixture was diluted with a mixture of NMP, ethanol, and acetic acid, filtered, and purified by preparative HPLC (10-50% acetonitrile in water containing 0.1% TFA). The fractions containing the desired product were combined and freeze-dried to dryness to obtain compound I-11 as a white solid. Yield: 14.9 mg, 54%; LC-MS m / z 636.4[M+1]+; 1 H NMR (300MHz, DMSO-d6 containing D2O) δ7.75(s,1H),4.57(s,1H),4.51-4.35(m,2H),3.84-3.65(m,5H),3.60-3.45(m,2H),3.41-3.29(m,1H),3 .21(t,J=9.3Hz,1H),3.15-3.03(m,1H),3.03-2.88(m,2H),2.88-2.74(m,2H),2.02-1.82(m,1H),1.79-1.59(m,1H),1.56-1.28(m,2H).

[0622] Example 12: Synthesis of Compound I-4 [ka]

[0623] N-(acid-PEG3)-N-bis(PEG3-azide)(12A) (1.00 equivalent, 18.3 mg, 0.0293 mmol) and N,N'-dicyclohexylcarbodiimide (DCC) (1.00 equivalent, 6.1 mg, 0.0293 mmol) were dissolved in NMP (0.1 mL) with stirring. After 5 minutes, a solution of 2,3,4,5,6-pentafluorophenol (1.50 equivalent, 8.1 mg, 0.0440 mmol) in NMP (0.2 mL) was added. The resulting clear solution was covered and stirred at room temperature for 2 hours, at which point a catalytic amount of DMAP was added (a white precipitate slowly formed). After 16 hours and 23 hours, DCC (3 mg + 1 mg) was added further. After 24 hours, the resulting mixture was added to compound 8D (2.00 equivalents, 18.2 mg, 0.0587 mmol) in a drum vial equipped with a stirring bar. After 2 minutes, tetrakis(acetonitrile)copper(I) hexafluorophosphate (5.00 equivalents, 54.7 mg, 0.147 mmol) was added. The resulting pale yellow solution was capped and stirred at room temperature for 30 minutes. The reaction mixture was diluted with a mixture of NMP, ethanol, and acetic acid, filtered, and purified by preparative HPLC (10-40% acetonitrile in water containing 0.1% TFA). The fractions containing the desired product were combined and lyophilized to dryness to obtain compound I-12 as a white solid. Yield: 8.7 mg, 21%; LC-MS m / z 1410.9[M+1] + ; 1 H NMR (300MHz, DMSO-d6 containing D2O) δ7.81(s,2H),4.60(s,2H),4.45(s,4H),3.87-2.76(m,50H),2.03-1.83(m,2H),1.79-1.59(m,2H),1.55-1.29(m,4H).

[0624] Example 13: Synthesis of Compound I-13 [ka]

[0625] A solution of azide-PEG1-amine (13A) (1.30 equivalents, 8.5 mg, 0.0649 mmol) in NMP (0.3000 mL) was added to compound 10A (1.30 equivalents, 17.3 mg, 0.0649 mmol) in a drum vial equipped with a stirring rod. The resulting colorless, transparent solution was capped and stirred at room temperature for 30 minutes, then added to compound 8D (1.00 equivalents, 15.5 mg, 0.0500 mmol) in a drum vial equipped with a stirring rod. After 2 minutes, tetrakis(acetonitrile)copper(I) hexafluorophosphate (2.80 equivalents, 52.1 mg, 0.140 mmol) was added. The resulting pale yellow solution was capped and stirred at room temperature for 30 minutes. The reaction mixture was diluted with a mixture of NMP, ethanol, and acetic acid, filtered, and purified by preparative HPLC (5-30% acetonitrile in water containing 0.1% TFA). The fractions containing the desired product were combined and freeze-dried to obtain compound I-13 as a white solid. Yield: 15.0 mg, 51%; LC-MS m / z 592.4[M+1]+; 1 H NMR (300MHz, DMSO-d6 containing D2O) δ7.81(s,1H),6.95(s,2H),4.60(s,1H),4.52-4.36(m,2H),3.80-3.51(m,6H),3.42-3.29( m,3H),3.27-3.03(m,5H),2.91-2.78(m,2H),2.37-2.23(m,2H),2.01-1.85(m,1H),1.79-1.60(m,1H),1.54-1.33(m,2H).

[0626] Example 14: Synthesis of Compound I-14 [ka]

[0627] A solution of azide-PEG7-amine (14A) (1.00 equivalent, 16.9 mg, 0.0429 mmol) in NMP (0.3000 mL) was added to compound 10A (1.00 equivalent, 11.4 mg, 0.0429 mmol) in a drum vial equipped with a stirring rod. The resulting colorless, transparent solution was capped and stirred at room temperature for 30 minutes, then added to compound 8D (1.00 equivalent, 13.3 mg, 0.0429 mmol) in a drum vial equipped with a stirring rod. After 2 minutes, tetrakis(acetonitrile)copper(I) hexafluorophosphate (2.80 equivalents, 44.7 mg, 0.120 mmol) was added. The resulting pale yellow solution was capped and stirred at room temperature for 30 minutes. The reaction mixture was diluted with a mixture of NMP, ethanol, and acetic acid, filtered, and purified by preparative HPLC (5-35% acetonitrile in water containing 0.1% TFA). The fractions containing the desired product were combined and freeze-dried to obtain compound I-14 as a white solid. Yield: 8.6 mg, 23%; LC-MS m / z 856.5[M+1]+; 1 H NMR (300MHz, DMSO-d6 containing D2O) δ8.04(bs,1H),7.83(s,1H),6.97(s,2H),4.60(s,1H),4.52-4.38(m,2H),3.84-3.66(m,4H),3.52-3.2 8(m,29H),3.28-3.04(m,5H),2.85(t,J=6.7Hz,2H),2.31(t,J=7.4Hz,2H),2.03-1.86(m,1H),1.80-1.60(m,1H),1.56-1.29(m,2H).

[0628] Example 15: Synthesis of Compound I-15 [ka]

[0629] A solution of azide-PEG11-amine (15A) (1.00 equivalent, 24.8 mg, 0.0435 mmol) in NMP (0.3000 mL) was added to compound 10A (1.00 equivalent, 11.6 mg, 0.0435 mmol) in a drum vial equipped with a stirring rod. The resulting colorless, transparent solution was capped and stirred at room temperature for 30 minutes, then added to compound 8D (1.00 equivalent, 13.5 mg, 0.0435 mmol) in a drum vial equipped with a stirring rod. After 2 minutes, tetrakis(acetonitrile)copper(I) hexafluorophosphate (2.80 equivalents, 45.4 mg, 0.122 mmol) was added. The resulting pale yellow solution was capped and stirred at room temperature for 30 minutes. The reaction mixture was diluted with a mixture of NMP, ethanol, and acetic acid, filtered, and purified by preparative HPLC (5-35% acetonitrile in water containing 0.1% TFA). The fractions containing the desired product were combined and freeze-dried to dryness to obtain compound I-15 as a colorless semi-solid. Yield: 17.2 mg, 38%; LC-MS m / z 1032.6[M+1]+; 1 H NMR (300MHz, DMSO-d6 containing D2O) δ7.81(s,1H),6.91(s,2H),4.59(s,1H),4.50-4.36(m,2H),3.91-3.65(m,19H),3.62-3.27(m, 30H),3.27-3.03(m,5H),2.91-2.78(m,2H),2.30(t,J=7.4Hz,2H),1.99-1.85(m,1H),1.80-1.60(m,1H),1.55-1.33(m,2H).

[0630] Example 16: Synthesis of Compound I-16 [ka]

[0631] To a solution of perfluorophenyl 3-(2-(2-azidoethoxy)ethoxy)propanoate (16A) (1.0 equivalent, 0.200 g, 0.542 mmol) in dimethyl sulfoxide (4 mL), 3-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)-N-(3,6,9,12-tetraoxapentadeca-14-in-1-yl)propanamide (16B) (1.5 equivalents, 0.311 g, 0.812 mmol) was added and the mixture was stirred for 5 minutes. Then, tetrakis(acetonitrile)copper(I) hexafluorophosphate (2.8 equivalents, 0.565 g, 1.52 mmol) was added, and the reaction mixture was stirred at room temperature for 1 hour. After completion, the reaction mixture was diluted with acetonitrile and purified by preparative HPLC (45-75% acetonitrile in water containing 0.1% TFA). The fractions containing the desired product were combined and freeze-dried to obtain compound 16C as a colorless viscous liquid. Yield: 0.045 g, 10.88%; LC-MS m / z 752.33 [M+1] + .

[0632] In dimethyl sulfoxide (0.6 mL), a molecular sieve (powder, catalyst support, sodium Y zeolite, Aldrich catalog number 334448) was added, followed by intermediate A-10 (1.0 equivalent, 0.019 g, 0.054 mmol), triethylamine (3.0 equivalent, 0.023 mL, 0.163 mmol), and compound 16C (1.1 equivalent, 0.045 g, 0.059 mmol). The reaction mixture was stirred at room temperature for 3 hours. After completion, the reaction mixture was diluted with acetonitrile and purified by preparative HPLC (13-23% acetonitrile in water containing 0.1% TFA). The fractions containing the desired product were combined and freeze-dried to dryness to obtain compound I-16 as an off-white solid. Yield: 0.008 g, 15.82%; LC-MS m / z 917.33 [M+1] + ; 1H NMR(400MHz,D2O)δ7.98(s,1H),7.37(d,J=8.8Hz,2H),7.13(d,J=8.8Hz,2H),6.83(s,2H),5.55 (s,1H),4.61(s,2H),4.56-4.54(m,2H),4.17-4.16(m,1H),4.00-3.98(m,1H),3.94(t,J=4.8Hz, 2H),3.82-3.75(m,4H),3.68-3.58(m,18H),3.53(t,J=5.2Hz,2H),3.29(t,J=5.6Hz,2H),2.64( t,J=6.0Hz,2H),2.48(t,J=6.4Hz,2H),2.15-1.90(m,1H),1.80-1.60(m,2H),1.40-1.25(m,1H).

[0633] Example 17: Synthesis of Compound I-17 [ka]

[0634] Compound I-17 is synthesized using the procedure described for compound I-7, but with 1-(14-azido-3,6,9,12-tetraoxatetradecyl)-1H-pyrrole-2,5-dione (17A) instead of compound 7B.

[0635] Example 18: Synthesis of Compound I-18 [ka]

[0636] Compound I-18 is synthesized using the procedure described for compound I-7, but with 1-(14-azido-3,6,9,12-tetraoxatetradecyl)-3,4-dibromo-1H-pyrrole-2,5-dione (18A) instead of compound 7B.

[0637] Example 19: Synthesis of intermediates XA, XB, and XC [ka]

[0638] Intermediate XA is synthesized using the procedure described for intermediate A, with compound XH as the starting material instead of mannose 6-phosphate. [ka]

[0639] Intermediate XB is synthesized using the procedure described for intermediate A-10, with XH as the starting material instead of mannose 6-phosphate. [ka]

[0640] Intermediate XC is synthesized using the procedure described for compound 8D, with XH as the starting material instead of mannose 6-phosphate.

[0641] Example 20 [ka]

[0642] Compound 20B is synthesized by using compound 20A instead of compound 1A, and following the procedure described for compound 1B.

[0643] Compound I-20 is synthesized using the procedure described for Compound 1, but with Compound 20B and intermediate XB instead of Compound 1B and intermediate A-10.

[0644] Example 21 [ka]

[0645] Compound 21B is synthesized by using compound 21A and pentafluorophenol instead of compound 1A and 2,3,5,6-tetrafluorophenol, using the procedure described for compound 1B.

[0646] Compound I-21 is synthesized using the procedure described for Compound 1, but with Compound 21B and intermediate XB instead of Compound 1B and intermediate A-10.

[0647] Example 22 [ka]

[0648] Compound 22B is synthesized by using compound 22A and pentafluorophenol instead of compound 1A and 2,3,5,6-tetrafluorophenol, using the procedure described for compound 1B.

[0649] Compound I-22 is synthesized using the procedure described for Compound 1, but with Compound 22B and intermediate XB instead of Compound 1B and intermediate A-10.

[0650] Example 23 [ka]

[0651] Compounds 23B and 23C are synthesized by using compounds 23A and 23B instead of compounds 2C and 2D, and following the procedure described for compounds 2D and 2E.

[0652] Compound I-23 is synthesized using the procedure described for compound 2, but with compound 23C and intermediate XB instead of compound 2E and intermediate A-10.

[0653] Example 24 [ka]

[0654] Compounds 24B and 24C are synthesized using the same procedure described for compounds 2D and 2E, but with compounds 24A, 23A, and 24B instead of compounds 2B, 2C, and 2D.

[0655] Compound I-24 is synthesized using the procedure described for Compound 2, but with Compound 24C and intermediate XB instead of Compound 2E and intermediate A-10.

[0656] Example 25 [ka]

[0657] Compound I-25 is synthesized using the procedure described for compound I-6, but with compound 25A and intermediate XB instead of compound 6A and intermediate A-10.

[0658] Example 26 [ka]

[0659] Compound I-26 is synthesized using the procedure described for compound I-13, but with compound 26A and intermediate XA instead of compounds 13A, 8D, and tetrakis(acetonitrile)copper(I)hexafluorophosphate.

[0660] Example 27 [ka]

[0661] Compound I-27 is synthesized using the procedure described for compound I-13, but with compound 27A and intermediate XA instead of compounds 13A, 8D, and tetrakis(acetonitrile)copper(I)hexafluorophosphate. Deprotection of the Boc protecting group is carried out under standard Boc deprotection conditions before adding intermediate XA.

[0662] Example 28 [ka]

[0663] Compound I-28 is synthesized using the procedure described for compound I-13, but with compound 28A and intermediate XA instead of compounds 13A, 8D, and tetrakis(acetonitrile)copper(I)hexafluorophosphate. Deprotection of the Boc protecting group is carried out under standard Boc deprotection conditions before adding intermediate XA.

[0664] Example 29 [ka]

[0665] Compound 29B is synthesized by using compound 29A and intermediate XB instead of compound 5A and intermediate A-10, using the procedure described for compound 5B.

[0666] Compound I-29 is synthesized using the procedure described for compound I-5, but with compounds 29B and 29C instead of compounds 5B and 5C.

[0667] Example 30 [ka]

[0668] Compound 30B is synthesized by using compound 30A instead of compound 12A, and following the procedure described for compound 12B.

[0669] Compound I-30 is synthesized using the procedure described for compound I-12, but with compound 30B and intermediate XC instead of compounds 12B and 8D.

[0670] Example 31 [ka]

[0671] Compound 31B is synthesized by using compound 31A instead of compound 12A, and following the procedure described for compound 12B.

[0672] Compound I-31 is synthesized using the procedure described for compound I-12, but with compound 31B and intermediate XC instead of compounds 12B and 8D.

[0673] Example 32 [ka]

[0674] Compound 32B is synthesized by using compound 32A instead of compound 12A, and following the procedure described for compound 12B.

[0675] Compound I-32 is synthesized using the procedure described for compound I-12, but with compound 32B and intermediate XC instead of compounds 12B and 8D.

[0676] Example 33: Synthesis of Compound I-33 [ka]

[0677] DBU (0.1 equivalent) is added at 0°C under nitrogen to a stirred solution of dibenzyl (2-((2R,3R,4S,5S,6S)-3,4,5-tris(benzyloxy)-6-hydroxytetrahydro-2H-pyran-2-yl)ethyl)phosphonate (33A) (1.00 equivalent) and trichloroacetonitrile (10.0 equivalent) in DCM. The resulting mixture is stirred at 0°C under nitrogen until LC-MS shows complete conversion to compound 33B. Most of the solvent is removed using a rotary evaporator. The residue is purified by silica gel chromatography to obtain compound 33B. Compound 33B (1.00 equivalent) is dissolved in dry DCM with stirring under nitrogen. Add perfluorophenyl 14-hydroxy-3,6,9,12-tetraoxatetradecanoate (33C) (2.00 equivalents), and cool the resulting mixture to -78°C while stirring under nitrogen. Slowly add a solution of boron trifluoride diethyl ether (0.500 equivalents) in dichloromethane. Remove the -78°C cooling bath, and slowly warm the reaction mixture to 0°C under nitrogen, then work up. Purify the crude product by silica gel chromatography to obtain compound 33D. Dissolve compound 33D (1 equivalent) in dry ethyl acetate while stirring. Add palladium carbon (0.05 equivalents), and vigorously stir the resulting mixture under a hydrogen balloon until LC-MS shows complete conversion to compound I-33. Filter the resulting mixture through Celite, concentrate it in a rotary evaporator, and purify it by reverse-phase preparative HPLC to obtain compound I-33.

[0678] Example 34: Synthesis of Compound I-34 [ka]

[0679] Compound 33B (1.00 equivalent) is dissolved in dry DCM while stirring under nitrogen. Perfluorophenyl 14-hydroxytetradecanoate (34A) (2.00 equivalent) is added, and the resulting mixture is cooled to -78°C while stirring under nitrogen. A solution of boron trifluoride diethyl ether (0.500 equivalent) in dichloromethane is slowly added. The -78°C cooling bath is removed, and the reaction mixture is slowly warmed to 0°C under nitrogen, followed by workup. The crude product is purified by silica gel chromatography to obtain compound 34B. Compound 34B (1 equivalent) is dissolved in dry ethyl acetate while stirring. Palladium carbon (0.05 equivalent) is added, and the resulting mixture is vigorously stirred under a hydrogen balloon until LC-MS shows complete conversion to compound I-34. The resulting mixture is filtered through Celite, concentrated in a rotary evaporator, and purified by reverse-phase preparative HPLC to obtain compound I-34.

[0680] Example 35: Synthesis of Compound I-35 [ka]

[0681] Compound 33B (1.00 equivalent) is dissolved in dry DCM with stirring under nitrogen. Perfluorophenyl 8-(2-(2-hydroxyethoxy)ethoxy)octanoate (35A) (2.00 equivalent) is added, and the resulting mixture is cooled to -78°C with stirring under nitrogen. A solution of boron trifluoride diethyl ether (0.500 equivalent) in dichloromethane is slowly added. The -78°C cooling bath is removed, and the reaction mixture is slowly warmed to 0°C under nitrogen, followed by workup. The crude product is purified by silica gel chromatography to obtain compound 35B. Compound 35B (1 equivalent) is dissolved in dry ethyl acetate with stirring. Palladium carbon (0.05 equivalent) is added, and the resulting mixture is vigorously stirred under a hydrogen balloon until LC-MS shows complete conversion to compound I-35. The resulting mixture was filtered through Celite, concentrated using a rotary evaporator, and purified by reverse-phase preparative HPLC to obtain compound I-35.

[0682] Example 36: Synthesis of Compound B [ka]

[0683] Compound B is synthesized using the procedure described for compound 8D, with buto-3-in-1-amine instead of buto-3-in-1-ol.

[0684] Alternatively, intermediate B-2 can be prepared by adding pyridine to a solution of intermediate B-1 in excess acetic anhydride. The resulting mixture is stirred at 20°C for 16 hours. The reaction solution is concentrated under vacuum, and the remaining pyridine is removed by azeotropic distillation with toluene, followed by high-vacuum drying, to obtain intermediate B-2.

[0685] Example 37: Synthesis of Compound I-37 [ka]

[0686] Compound I-37 is synthesized using compound B instead of compound 8D, following the procedure described for compound I-8.

[0687] Example 38: Synthesis of Compound I-38 [ka]

[0688] To a round-bottom flask containing intermediate A-8 (1.00 equivalent, 218 mg, 0.398 mmol), (4-nitrophenyl)N-hexa-5-inylcarbamate (38A) (1.80 equivalent, 188 mg, 0.717 mmol) and anhydrous DCM (4 mL) were added. Triethylamine (2.08 equivalent, 0.11 mL, 0.826 mmol) was added to the reaction solution, and the solution was stirred at 40°C for 16 hours. The reaction mixture was then diluted with dichloromethane (30 mL) and washed with aqueous NaOH, water, and brine. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography and eluted with methanol / chloroform to obtain compound 38B. Yield: 154 mg, 58%); LCMS m / z 655.6[M+1]+.

[0689] Acetonitrile (4 mL) was added to a nitrogen-purged round-bottom flask containing compound 38B (1.00 equivalent, 170 mg, 0.260 mmol). The solution was cooled to 0°C under nitrogen before dropwise addition of TMSBr (5.00 equivalent, 0.18 mL, 1.30 mmol). The cooling bath was removed, and the resulting mixture was stirred at room temperature under nitrogen. LC-MS after 2 hours showed no residual SM and the product M+H=599.6 was observed. The solvent was removed using a rotary evaporator, and the residue was dried under high vacuum. The obtained intermediate, 2-[(2R,3R,4S,5S,6R)-3,4,5-triacetoxy-6-[4-(hexa-5-inylcarbamoylamino)phenoxy]tetrahydropyran-2-yl]ethylphosphonic acid (155 mg, 0.259 mmol, yield 99.72%), was dissolved in methanol (3 mL). 25 wt% NaOMe in MeOH (2.50 equivalents, 0.14 mL, 0.649 mmol) was added to the stirred solution under nitrogen. The resulting mixture was stirred under nitrogen at room temperature for 50 minutes. LC-MS showed that most of the starting material remained. Another aliquot of 25 wt% NaOMe in MeOH (2.50 equivalents, 0.14 mL, 0.649 mmol) was added, and the mixture was stirred at 20°C for at least 1 hour. Acetic acid (13.5 equivalents, 0.20 mL, 3.50 mmol) was added, and the solvent was removed using a rotary evaporator. The residue was taken up in DMSO and purified by preparative HPLC (0-35% acetonitrile in water containing 0.1% TFA). The purified product fractions were combined and freeze-dried to dryness to obtain compound 38C as a white solid. Yield: 45 mg, 37%; LCMS m / z 473.6[M+1]+.

[0690] Compound 38C (1.00 equivalent, 19.0 mg, 0.0402 mmol) was placed in a nitrogen-purged glass vial equipped with a stirring rod. A solution of compound 7B (1.20 equivalent, 22.1 mg, 0.0483 mmol) in NMP (1 mL) was added to the vial, followed by [(CH3CN)4Cu]PF6 (2.50 equivalent, 37.5 mg, 0.101 mmol). The resulting clear yellow solution was capped and stirred at room temperature for 30 minutes. LC-MS analysis confirmed that the reaction was complete. The reaction mixture was diluted with a mixture of NMP, ethanol, and acetic acid, filtered, and purified by preparative HPLC (15-65% acetonitrile in water containing 0.1% TFA) over a 30-minute run. The fractions containing the desired product were combined and lyophilized to dryness to obtain compound I-38 as a white solid. Yield: 12 mg, 37%; LCMS m / z 930.5[M+1]+; 1 H NMR(300MHz,DMSO-d6 D2O)δ7.77(s,1H),7.24(d,J=8.5Hz,2H),6.88(d,J=8.6Hz,2H),5.23(s,1H),4.42(t,J=5.1Hz,2H),3.89-3.25(m,20H),3. 05(t,J=6.2Hz,2H),2.95(t,J=5.8Hz,2H),2.59(t,J=7.5Hz,2H),2.02-1.82(m,1H),1.70-1.33(m,6H),1.30-1.05(m,1H).

[0691] Example 39: Synthesis of Compound I-39 [ka]

[0692] Octo-7-inoic acid (1.66 equivalents, 82.6 mg, 0.589 mmol), DMF (3 mL), and HATU (1.50 equivalents, 203 mg, 0.534 mmol) were added to a nitrogen-purged round-bottom flask. The reaction solution was stirred at 20°C for 20 minutes before adding intermediate A-8 (1.00 equivalents, 195 mg, 0.356 mmol) in 1 mL of DMF. The reaction solution was stirred at 20°C for 24 hours before analysis by LC-MS. The reaction solution was diluted with HCl (30 mL) and washed with saturated NH4Cl aqueous solution (20 mL), followed by saturated NaCl aqueous solution (20 mL). The partitioned HCl phase was dried over Na2SO4, filtered, and concentrated under vacuum to obtain the crude product, which was purified over 15 minutes by silica gel column chromatography using a mobile phase of 100% Hx to 75% HCl / Hx to obtain compound 39A. Yield: 182 mg, 76%; LCMS m / z 653.6[M+1]+.

[0693] In a nitrogen-purged round-bottom flask containing compound 39A (1.00 equivalent, 182 mg, 0.278 mmol) and anhydrous acetonitrile (1 mL), TMSBr (5.00 equivalent, 0.18 mL, 1.39 mmol) was added at 0°C under nitrogen. After removing the cooling bath, the resulting mixture was stirred under nitrogen at room temperature for 3.5 hours. LC-MS analysis showed that no starting reagent remained. Volatile substances were removed using a rotary evaporator, and the residue was dried briefly under high vacuum. The residue was dissolved in methanol (1 mL) while stirring under nitrogen, and 25% by weight of sodium methoxide in MeOH (2.50 equivalent, 0.15 mL, 0.696 mmol) was added. The resulting mixture was stirred under nitrogen at room temperature for 30 minutes. Acetic acid (5.00 equivalent, 0.080 mL, 1.39 mmol) was added to the reaction mixture, and volatile substances were removed under vacuum. The residue was incorporated into DMSO and purified by reverse-phase preparative HPLC (0-35% acetonitrile in water containing 0.1% TFA) to obtain a purified fraction. The combined fraction was freeze-dried to dryness to obtain compound 39B as a white solid. Yield: 65 mg, 50%; LCMS m / z 472.3[M+1]+

[0694] Compound 39B was added to a nitrogen-purged glass vial equipped with a magnetic stirring rod. A solution of compound 7B (1.20 equivalents, 34.9 mg, 0.0764 mmol) in NMP (1 mL) was added to the vial, followed by [(CH3CN)4Cu]PF6 (2.50 equivalents, 59.3 mg, 0.159 mmol). The resulting clear yellow solution was capped and stirred at room temperature for 30 minutes. LC-MS analysis confirmed that no starting material remained. The reaction mixture was diluted with a mixture of NMP (0.3 mL), ethanol (0.3 mL), and acetic acid (0.3 mL), filtered, and purified by preparative HPLC (15-65% acetonitrile in water containing 0.1% TFA) to obtain a purified fraction. The fractions containing the desired product were combined and freeze-dried to dryness to obtain compound I-39 as a white solid. Yield: 55 mg, 59%; LCMS m / z 929.6[M+1]+; 1 H NMR (300MHz, DMSO-d6 containing D2O) δ7.76(s,1H),7.46(d,J=8.8Hz,2H),6.94(d,J=8.2Hz,2H),5.28(s,1H),4.41(t,J=5.1Hz,2 H),3.86-2.87(m,22H),2.64-2.53(m,2H),2.23(t,J=7.5Hz,2H),1.99-1.80(m,1H),1.68-1.40(m,6H),1.37-1.05(m,3H).

[0695] Example 40: Synthesis of Compound I-40 [ka]

[0696] To a stirred solution of compound 12A (1.00 equivalent, 500 mg, 0.802 mmol) in THF (2.5 mL), DCC (1.50 equivalent, 248 mg, 1.20 mmol), a solution of 2,3,4,5,6-pentafluorophenol (1.70 equivalent, 251 mg, 1.36 mmol) in THF (1 mL), and then 4-dimethylaminopyridine (0.0300 equivalent, 2.9 mg, 0.0241 mmol) were added sequentially. The resulting mixture was capped and stirred at room temperature for 17 hours. The reaction mixture was diluted with Et2O and filtered. The filtrate was concentrated using a rotary evaporator. The residue was taken into DCM and purified by silica gel chromatography (0-100% acetonitrile in DCM) to obtain compound 12B as a yellow oil. Yield: 258 mg, 41%; LCMS m / z 790.7[M+1]+; 1 ¹H NMR (300MHz, chloroform-d) δ 3.87 (t, J=6.2Hz, 2H), 3.74-3.56 (m, 16H), 3.39 (t, J=5.1Hz, 4H), 2.94 (t, J=6.2Hz, 2H).

[0697] A solution of compound 40A (2.20 equivalents, 36.1 mg, 0.0738 mmol) in NMP (0.6 mL) was added to compound 12B (1.00 equivalent, 26.5 mg, 0.0336 mmol) in a drum vial equipped with a stirring bar. The resulting solution was stirred, and [(CH3CN)4Cu]PF6 (5.00 equivalents, 62.5 mg, 0.168 mmol) was added. The resulting pale yellow solution was capped and stirred at room temperature for 25 minutes. The reaction mixture was diluted with a mixture of NMP, ethanol, and acetic acid, filtered, and purified by preparative HPLC (15-40% acetonitrile in water containing 0.1% TFA). The fractions containing the desired product were combined and lyophilized to dryness to obtain compound I-40 as a white solid. Yield: 38.9 mg, 66%; LCMS m / z 1765.9[M-1]-; 1H NMR (300MHz, DMSO-d6 containing D2O) δ7.81(s,2H),7.19(d,J=8.5Hz,4H),6.99(d,J=8.8Hz,4H),5.33(s,2H),4.43(t,J=5.2Hz,4H) ),3.90-3.23(m,54H),2.97(t,J=5.8Hz,2H),2.69-2.34(m,4H),2.01-1.81(m,2H),1.73-1.40(m,12H),1.34-1.10(m,2H).

[0698] Example 41: Synthesis of Compound I-41 [ka]

[0699] Compound I-40 (1.00 equivalent, 32.7 mg, 0.0185 mmol) in a vial equipped with a stirring bar was mixed with a solution of 1-(2-aminoethyl)-1H-pyrrole-2,5-dione TFA salt (1.15 equivalent, 5.4 mg, 0.0213 mmol) and DIPEA (3.00 equivalent, 0.0097 mL, 0.0555 mmol) in NMP (1 mL). The resulting clear, pale yellow solution was capped and stirred at room temperature for 30 minutes. The reaction mixture was diluted with acetic acid, filtered, and purified by preparative HPLC (10-30% acetonitrile in water containing 0.1% TFA). The fractions containing the desired product were combined and lyophilized to dryness to obtain compound I-41 as a pale yellow solid. Yield: 18.5 mg, 58%; LCMS m / z 1722.0[M-1]-; 1 H NMR (300MHz, DMSO-d6 containing D2O) δ7.82(s,2H),7.26-7.09(m,4H),7.00(d,J=8.5Hz,4H),6.90(s,2H),5.33(s,2H),4.52-4.32(m ,4H),3.99-2.94(m,58H),2.69-2.57(m,4H),2.20(t,J=6...

Claims

[Claim 1] The invention described in the present specification.

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