Cleavable conjugates of transthyretin protein ligands and uses thereof

Abstract

Provided herein are conjugates of Formula (I): (I), or a pharmaceutically acceptable salt thereof, wherein T, L, L B , E and D 1 are as defined elsewhere herein. Also provided are methods of making compounds of Formula (I).

Description

Cross-reference to related applications

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 572,081, filed March 29, 2024, and U.S. Provisional Application No. 63 / 748,386, filed January 22, 2025, the entire contents of each of which are incorporated herein by reference. Technical Field

[0002] Aspects of the present invention generally relate to an active agent delivery system comprising: a ligand (binding agent) selective for transthyretin (TTR) in the serum of a subject; and a cleavable connector configured to operatively covalently link the ligand to a drug to increase the plasma circulation time and duration of the drug in the serum. Background Technology

[0003] Adverse pharmacokinetic profiles of many drugs represent a significant challenge in their continued development. This problem, in particular, limits a drug's ability to achieve its impressive therapeutic potential. Specifically, the half-life of many drugs in the serum of subjects is of great concern. A short half-life is generally attributed to (i) enzymatic degradation by serum proteases and / or (ii) rapid renal clearance due to the relatively high molecular weight cutoff (approximately 30,000 Daltons) of peptides and proteins to be cleared via glomerular filtration. Those skilled in the art recognize that, for example, the short in vivo half-life of many drug candidates limits their clinical potential by increasing the dose size and frequency required to achieve the desired results.

[0004] Therefore, the art should understand a delivery system that (i) improves the duration of action and plasma circulation of the active drug; (ii) avoids steric hindrance problems that affect the binding affinity and potency of the active agent; (iii) avoids renal tubular vacuolation caused by PEGylation; (iv) avoids the generation of antibodies that cause immune response problems; and (v) has low manufacturing cost. Summary of the Invention

[0005] It has been unexpectedly discovered that the duration of action (DOA) of a drug can be significantly prolonged by covalently linking the drug to a ligand selective for transthyretin (TTR) via a cleavable linker. Therefore, this disclosure relates to a novel prodrug method for enhancing plasma circulation and DOA of a drug without affecting its biological activity.

[0006] In one respect, this paper provides a conjugate of formula (I): Or its pharmaceutically acceptable salt, wherein: T is a ligand selective for transthyretin-TTR; L stands for connector; L B Click products that do not exist or are formed through a click reaction between the first click handle and the second click handle; E is a cleavable ester; and D 1 This refers to the pharmaceutical component.

[0007] In some embodiments, the cleavable ester moiety (E) of the formula (I) conjugate comprises an aromatic ring. In some embodiments, the aromatic ring may optionally be substituted with one or more substituents as set forth herein. In some embodiments, p-aminobenzoic acid (PABA) is used as the basic structural unit for forming an ester with the drug. In some embodiments, p-hydroxybenzoic acid (PHBA) is used as the basic structural unit for forming an ester with the drug moiety. In some embodiments, substituted PABA is used as the basic structural unit for forming an ester with the drug. In some embodiments, the aromatic ester group (e.g., PABA or PHBA) of the formula (I) conjugate is hydrolyzed by carboxylesterase 2 (CES2) to a greater extent than by carboxylesterase 1 (CES1). In some embodiments, the aromatic ester group (e.g., PABA or PHBA) of the formula (I) conjugate is hydrolyzed by CES2, but only minimally by CES1. In some embodiments, the aromatic ester group (e.g., PABA) of the formula (I) conjugate is completely resistant to CES1.

[0008] Surprisingly, in some embodiments, TTR ligands have been found to influence the degree of hydrolysis of aromatic ester groups (e.g., PABA or PHBA groups) in the conjugate and the selectivity of carboxylesterases. For example, in some embodiments, the TTR ligand (binding agent) of the conjugate of formula (I) is able to increase the CES2 selectivity of the conjugate of formula (I). This CES2 selectivity provides a more controlled or prolonged release of the drug from the conjugate. In addition, the hydrolysis rate can be effectively modulated (e.g., increased or decreased) by specific substitutions on the aromatic ester groups (e.g., PABA or PHBA groups) of the conjugate.

[0009] In some embodiments, the formula (I) conjugate comprises the following ester moiety: in Indicates with L or L B The connection point.

[0010] In some embodiments, the formula (I) conjugate comprises the following ester moiety: in Indicates with L or L B The connection point, R1x For H, F, Cl, NO2, CF3 or OMe, and R 1y For H or Me. In some implementations, R 1x For H or Me and R 1y It can be H, F, Cl, Br, NO2, CF3, Me, or OMe. In some implementations, R 1x For H and R 1y For Me.

[0011] In some embodiments, the formula (I) conjugate comprises the following ester moiety: in Indicates with L or L B The connection point.

[0012] In some embodiments, the formula (I) conjugate comprises the following ester moiety: in Indicates with L or L B The connection point.

[0013] In some embodiments, the formula (I) conjugate comprises the following ester moiety: in Indicates with L or L B The connection point, R 1x For H, F, Cl, NO2, CF3 or OMe, and R 1y For H or Me. In some implementations, R 1x For H or Me and R 1y It can be H, F, Cl, Br, NO2, CF3, Me, or OMe. In some implementations, R 1x For H and R 1y For Me.

[0014] In some embodiments, the ligand selective for transthyretin protein TTR (T) is a compound of formula (E). (E) Or a pharmaceutically acceptable salt thereof, wherein variable R 1 R 2 R 3 R a R bX and a are as defined herein. In some embodiments, the formula (I) conjugate comprises a ligand of formula (E) and an aromatic ester group (e.g., PABA or PHBA). In some embodiments, the formula (E) compound increases the CES2 selectivity of the aromatic ester group (e.g., PABA or PHBA) within the compound.

[0015] In one embodiment, this document provides a conjugate of formula (II): (II), Or its pharmaceutically acceptable salt, wherein: Each R 1 Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n can be 0, 1, 2, 3, or 4; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4 alkyl; L stands for connector; Y 1 For key or C 1-4 Alkylene; Y 2 For NR 6 , O or S; R 6 For H or C 1-4 alkyl; Each R 7 Choose independently from the following groups: hydrogen, halogen atoms (e.g., Cl, Br, or F), NO2, CF3, OMe, O(C 1-4 Alkyl), hydroxyl, -OCF3, -NH2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C1-6 Alkoxy and C 3-6 cycloalkyl; p is 0, 1, 2, or 3; and D 1 This refers to the pharmaceutical component.

[0016] Any embodiment of the conjugate of formula (I) or formula (II) provided herein, or a pharmaceutically acceptable salt thereof, shall, where applicable, be applied to any other formula detailed herein, as if each embodiment were specifically and individually enumerated. Therefore, it should be understood and described that each embodiment of the conjugate of formula (I) or formula (II) provided herein is applicable to the conjugate of formulas (III) to (XII) or (III') to (VIII') or variants thereof, or a pharmaceutically acceptable salt thereof, as if each embodiment were specifically and individually enumerated. It should also be understood and described that all such embodiments may be used in any of the pharmaceutical compositions, methods, kits, uses, or other aspects detailed herein.

[0017] In one aspect, this document provides a method for increasing the plasma circulation time and DOA of a drug, said method comprising administering to a subject (e.g., a human patient) a therapeutically effective amount of a conjugate of this disclosure (e.g., a conjugate of formula (I) or (II)). In some embodiments, administration of a conjugate of this disclosure (e.g., a conjugate of formula (I) or (II)) reduces the immunogenicity of the active agent relative to administration of the drug in its free form (unconjugated form) alone.

[0018] In some embodiments, the conjugates of this disclosure are administered parenterally to a subject in need. For example, the conjugates of this disclosure may be administered subcutaneously, intravenously, intramuscularly, or intrathecally. In other embodiments, the conjugates of this disclosure are administered orally.

[0019] In some embodiments of the conjugates disclosed herein, administered subcutaneously, the conjugates form an accumulation in the subcutaneous tissue. In some such embodiments, the solubility of the conjugate at physiological pH is less than that of the unconjugated drug at physiological pH. Therefore, the slow dissolution of the conjugate relative to the unconjugated drug slows the release of the drug into systemic circulation, thus prolonging the duration of action of the drug. Subsequent conjugation of the conjugate with the TTR in systemic circulation further prolongs the duration of action of the drug. As shown in Example 11, in some embodiments, the ability of the conjugate to form an accumulation is attributed to the zwitterionic nature of the conjugate at physiological pH. In some embodiments, despite the reduced solubility of the conjugate relative to the unconjugated drug, the conjugate is rapidly absorbed, resulting in rapid action. Therefore, the conjugates of this disclosure provide a drug with rapid onset of action and prolonged duration of action.

[0020] Partly due to the balanced hydrophilicity of the TTR selective ligand, the conjugates of this disclosure (especially the (II) conjugate) exhibit enhanced tissue (particularly subcutaneous) absorption. Following tissue (e.g., subcutaneous) absorption, the reversible binding of the TTR selective ligand of the conjugate significantly prolongs the duration of action of the active agent. As illustrated herein, the cleavage of the linker between the TTR selective ligand and the active agent releases the active agent, thus allowing it to exert its pharmacological effects.

[0021] Furthermore, it has been found that the release of the active agent can be effectively controlled by appropriately selecting the cleavable moiety in the conjugate. For example, the specific ester moiety described herein undergoes slow hydrolysis after administration, thereby releasing the active agent in a controlled and sustained manner over a prolonged period of time. Slower hydrolysis and high binding to TTR effectively prolong plasma circulation time and duration of action (DOA).

[0022] Therefore, the conjugates of this disclosure exhibit several beneficial properties of novel prodrug methods. The balanced hydrophilicity of the conjugates allows for rapid absorption from tissues into the bloodstream, the binding of TTR prolongs plasma circulation time, and the cleavable moiety allows for modulated drug release. The desired release rate of a specific active drug from the conjugates of this disclosure will vary depending on the active agent.

[0023] In some implementation schemes, D 1 The peptide is present. As shown in the examples, the conjugates of this disclosure containing the peptide as an active pharmaceutical ingredient exhibit a significantly increased half-life duration of action relative to the free peptide pharmaceutical ingredient, thus enabling a reduction in the dosage of the peptide pharmaceutical ingredient.

[0024] In some embodiments, the peptide is RGK-MCA. In some embodiments, the peptide is neurotensin. In some embodiments, the peptide is gonadotropin-releasing hormone (GnRH). In some embodiments, the peptide is GIP (1-42) human peptide. In some embodiments, the peptide is D6-GnRH. In some embodiments, the peptide is insulin. In some embodiments, the peptide is teriparatide. In some embodiments, the peptide is calcitronin salmon. In some embodiments, the peptide is ipamorelin. In some embodiments, the peptide is vasopressin. In some embodiments, the peptide is glucagon. In some embodiments, the peptide is aclarastide. In some embodiments, the peptide is goserelin. In some embodiments, the peptide is leuprolide. In some embodiments, the peptide is teriparatide. In some embodiments, the peptide is lixisenatide. In some embodiments, the peptide is desmopressin. In some embodiments, the peptide is oxytocin. In some embodiments, the peptide is corticotropin. In some embodiments, the peptide is sermorelin. In some embodiments, the peptide is bremelanotide. In some embodiments, the peptide is abaloparatide. In some embodiments, the peptide is deslorelin.

[0025] In some embodiments, the peptide drug is a glucagon-like peptide-1 (GLP-1) receptor agonist used to treat obesity and / or diabetes. In some such embodiments, the peptide drug is semaglutide, liraglutide, dulaglutide, albiglutide, or exenatide. Because the duration of action of the peptide drug is significantly prolonged when incorporated into the conjugates of this disclosure, GLP-1 agonist conjugates can reduce the frequency of administration, such as once weekly, once every two weeks, once every three weeks, or once monthly.

[0026] In some embodiments, the peptide drug is a glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic peptide (GIP) receptor agonist used to treat obesity and / or diabetes. In some such embodiments, the peptide drug is tirzepatide. Because the duration of action of the peptide drug is significantly prolonged when incorporated into the conjugates of this disclosure, the GLP-1 / GIP agonist conjugates can reduce the frequency of administration, such as once weekly, once every two weeks, once every three weeks, or once monthly.

[0027] In some embodiments, the peptide is an endogenous opioid peptide. In some such embodiments, the peptide is endomorphin-1 or endomorphin-2. In other embodiments, the peptide is dermorphin.

[0028] In some embodiments, the active pharmaceutical ingredient (D) delivered via the conjugate of this disclosure 1 The active pharmaceutical ingredient (API) can be a small molecule, nucleotide, oligonucleotide, polynucleotide, amino acid, oligopeptide, polypeptide, or protein. It may include, but is not limited to, antiproliferative agents, antitumor agents, antimitotic agents, anti-inflammatory agents, antiplatelet agents, anticoagulants, antifibrinogens, antithrombins, antibiotics, antiallergens, and antioxidants.

[0029] In some implementation schemes, D 1 It can be peptides, oligopeptides, polypeptides, proteins, antibodies, oligonucleotides, polynucleotides, virus-like particles, small molecules, oligosaccharides, or imaging agents.

[0030] In some implementation schemes, D 1 For proteins. In some implementations, D 1 It is an antibody.

[0031] In some implementation schemes, D 1 The cytokine is used. In some embodiments, the cytokine is interleukin-2 (IL-2). In some embodiments, the cytokine is mutant IL-2, such as the cytokine described in US 9,206,943. In some embodiments, the cytokine is interleukin-15 (IL-15). In some embodiments, the cytokine is mutant IL-15. In one such embodiment, IL-15 is IL-15[N27D]. In some embodiments, the cytokine is interleukin-7 (IL-7). In some embodiments, the cytokine is interleukin-9 (IL-9). In some embodiments, the cytokine is interleukin-10 (IL-10). In some embodiments, the cytokine is interleukin-21 (IL-21). In some embodiments, the cytokine is interleukin-10 (IL-10).

[0032] In some implementation schemes, D 1 It is an anticancer agent. In some such implementations, D 1 For SN-38. In some such implementations, D 1 It is monomethyl auristatin E. In some such embodiments, D 1 For capecitabine. In some such implementations, D 1 It is irinotecan. In some such implementations, D 1 It is trifluridine. In some such embodiments, D 1 It is doxorubicin. In some such implementations, D... 1 For docetaxel. In some such implementations, D 1 It is infigratinib.

[0033] In some implementation schemes, D 1 It is an opioid antagonist. In some embodiments, the opioid antagonist is naloxone, nalmefene, naltrexone, methylnaltrexone, nalbuphine, or nalorphine.

[0034] In some implementation schemes, D 1 It is an opioid agonist. In some embodiments, the opioid agonist is morphine, oxycodone, hydromorphone, or methadone.

[0035] In some embodiments, the plasma circulation and duration of action (DOA) of the drug after administration of the conjugates of this disclosure (e.g., conjugates of formula (I) or (II)) is at least 12 hours. In some such embodiments, the DOA of the drug after administration of the conjugates of this disclosure (e.g., conjugates of formula (I) or (II)) is at least 20 hours. In some such embodiments, the DOA of the drug after administration of the conjugates of this disclosure is at least 96 hours. In some embodiments, the DOA of the drug after administration of the conjugates of this disclosure is at least 1 week. In some embodiments, the DOA of the drug after administration of the conjugates of this disclosure is at least 2 weeks. In some embodiments, the DOA of the drug after administration of the conjugates of this disclosure is at least 1 month.

[0036] In some embodiments, the conjugates disclosed herein can provide controlled and / or prolonged release of a drug. In some such embodiments, the drug can be safely administered from the slow release of the conjugate.

[0037] In some embodiments, the conjugates of this disclosure are administered subcutaneously to human subjects at a dose of about 0.005 mg / kg to about 40 mg / kg. In some embodiments, the conjugates of this disclosure are administered subcutaneously to human subjects at a dose of about 0.01 mg / kg to about 40 mg / kg. In some embodiments, the conjugates of this disclosure are administered subcutaneously to human subjects at a dose of about 10 mg / kg to about 40 mg / kg. In some embodiments, the conjugates of this disclosure are administered subcutaneously to human subjects at a dose of about 10 mg / kg to about 20 mg / kg. Attached Figure Description

[0038] Figure 1 illustrates how the prodrug of this disclosure selectively binds to TTR in human plasma and releases naloxone, other drugs, and peptides in rat plasma. Figure 1a The chemical structures of naloxone, AG10-L, and the prodrug are shown. AG10-L-E1-naloxone is an alkyl ester prodrug. AG10-L-E2-naloxone is a PABA aryl ester prodrug. C12-E2-naloxone is a control fatty acid-conjugated PABA aryl ester prodrug. Several other PABA-based prodrugs of small molecules and peptides are also shown. Figure 1b The water solubility of the compound (incubated at 100 μM) in potassium phosphate buffer (pH 7.4) and the logarithmic properties of the test compound in potassium phosphate buffer (pH 7.4) and n-octanol at physiological pH 7.4 were measured. D 7.4 value. Figure 1c The binding affinity of the compounds to purified human TTR in buffer was evaluated using fluorescence polarization assays. The compounds were tested at concentrations ranging from 0.01 μM to 20 μM. The Cheng-Prusoff equation was used, and IC50 was employed. 50 Value calculation combined with constant ( K d Data are expressed as mean ± sd (). n = 3 separate repetitions). Figure 1d The binding selectivity of the compound (tested at 10 μM) for TTR (approximately 5 μM) in human serum was evaluated. The modification of TTR in human serum by the covalent FPE probe was monitored for 6 h in the presence of the FPE probe alone (DMSO) or in the presence of the probe and the TTR ligand. Ligands exhibiting lower FPE probe fluorescence showed higher binding selectivity for TTR. Figure 1eThe percentage of occupancy was calculated using 3-hour readings for each individual probe. Data are expressed as mean ± SD. n = 4 separate repetitions). Figure 1f The plasma stability of the prodrugs AG10-L-E1-naloxone, AG10-L-E2-naloxone, and C12-E2-naloxone (as measured at 5 μM) in human plasma, and Figure 1g Plasma stability in rat plasma. The residual concentration in individual culture media was determined by HPLC at specified time points. Bar graphs represent the mean ± SD of the percentage of residual compound. n = 3 separate repetitions).

[0039] Figure 2 shows the bioactivation of AG10-L-E2-naloxone via CES2 enzyme hydrolysis. Figure 2a The chemical structures of AG10-L-E2-naloxone and its hydrolysis products AG10-L-E2-COOH and naloxone. Figure 2b AG10-L-E2-naloxone and human TTR (pdb id: 4HIQ) 6 Docking studies between AG10-L-E2-naloxone and mouse CES2 enzyme (pdb id: 8AXC). The linker length of AG10-L-E2-naloxone (approximately 29 Å) is long enough to allow AG10-L-E2-naloxone to bind to both CES2 and TTR simultaneously. Figure 2c AG10-L-E2-naloxone was hydrolyzed by purified CES2 in buffer but not by CES1. Figure 2d Metabolic bioactivation of AG10-L-E2-naloxone in liver microsomes from humans (HLM), rats (RLM), and monkeys (MLM). HLM efficiently generates naloxone from AG10-L-E2-naloxone. The selective CES2 inhibitor loperamide significantly reduced naloxone release, while the CES1 inhibitor digitonin had no effect. The concentration of naloxone released from AG10-L-E2-naloxone in HLM, MLM, and RLM was determined using a validated LC-MS / MS method. Figure 2e The stability of AG10-L-E2-naloxone (tested at 5 μM) in rat plasma in the absence and presence of the CES2 inhibitor loperamide and the CES1 inhibitor digitoxin. Figure 2f The stability of AG10-L-E2-naloxone (tested at 5 μM) in monkey plasma (plasma without CES1 or CES2) and rabbit plasma (plasma with high abundance of CES1 and CES2) was determined by HPLC at specified time points. Figures 2c to 2fThe bar chart represents the average ± sd of the remaining compound percentage. n = 3 separate repetitions).

[0040] Figure 3 shows the pharmacokinetic / pharmacodynamic evaluation of AG10-L-E2-naloxone in rats. Figure 3a Male Sporgdolly rats (each group) n = 3 Apply 15 µmol kg -1 The test compound (equivalent to 5 mg kg) -1 Naloxone and 15.8 mg / kg -1 A single subcutaneous (sc) injection of AG10-L-E2-naloxone was administered. The concentration of the test compound in plasma was determined at different time points and expressed as the mean ± sd of three biological replicates. Figure 3b brain concentration, and Figure 3c Naloxone in cerebrospinal fluid (CSF) (administered in free form) and naloxone released from AG10-L-E2-naloxone. Administered via a single subcutaneous (sc) injection of 15 µmol / kg. -1 The test compound (equivalent to 5 mg kg) -1 Naloxone and 15.8 mg / kg -1 AG10-L-E2-naloxone) in male Sporgodory rats (each group) n = 3 Naloxone concentrations in brain tissue and CSF were measured at 10 and 30 min after administration. Bar graphs show the individual mean ± sd of three biological replicates. Figure 3d Male Sporgdolly rats (per group) n = 9 Morphine overdose mortality data and antagonist rescue in a biologically replicated study. Morphine (subcutaneous dose of 100 mg / kg at 0 hr) -1 And at 6 hours, it was 50 mg / kg. -1 ) and antagonists (naloxone and AG10-L1-E2-naloxone; 15 µmol kg -1 The administration procedure is as follows: (a single subcutaneous dose at 5 min). Figure 3e Male Sporgdolly rats (per group) n = 8 Data on mortality from fentanyl overdose and rescue with antagonists in a biologically replicated study. Fentanyl (subcutaneous dose of 3 mg / kg at 0 hr) -1 And at 6 hours, it was 3 mg / kg. -1 ) and antagonists (naloxone and AG10-L1-E2-naloxone; 15 µmol kg -1 The administration procedure is as follows: (a single subcutaneous dose at 5 min). Figures 3f to 3iNaloxone, AG10-L-E1-naloxone, AG10-L-E2-naloxone, and C12-E2-naloxone (15 µmol kg) -1 (single subcutaneous dose at 1 min) in male Sporgodory rats (each group) n = 6 The reversal of fentanyl OIRD. Figure 3f Dosing regimens for fentanyl and its antagonists. Reversal of OIRD was expressed as minute ventilation relative to control (minute ventilation in rats prior to fentanyl injection was considered baseline). Figure 3g Minute ventilation (mean ± sd) was plotted as a function of time following the first dose of fentanyl and the antagonist; per group. n = 6 (Secondary biological repetition) Figure 3h The minute ventilation rate after the second fentanyl dose 6 hours later, and Figure 3i Minute ventilation was measured 24 hours after the third fentanyl dose. Animals were monitored in a plethysmography chamber 90 minutes after each fentanyl dose. Data were analyzed using GraphPad Prism V9.3.1.471 software (La Jolla, CA, USA).

[0041] Figure 4 illustrates the reversal of fentanyl OIRD by naloxone, nalmefene, and AG10-L-E2-naloxone. Figure 4a Male Sporgodory rats were administered 60 µmol / kg -1 The test compound (equivalent to 20 mg kg) -1 Naloxone and 63 mg kg -1 A single subcutaneous (sc) injection of AG10-L-E2-naloxone (per group) n = 3 The concentrations of the test compound in plasma were determined at different time points and expressed as the mean ± sd of three biological replicates. Figure 4b fentanyl and antagonist (60 µmol kg) -1 The dosing regimen was as follows. Reversal of OIRD was expressed as minute ventilation relative to the control (minute ventilation in rats prior to fentanyl injection was considered as baseline). Figure 4c Minute ventilation (mean ± sd) was plotted as a function of time following the first dose of fentanyl and the antagonist; per group. n = 6 (Secondary biological repetition) Figure 4d The minute ventilation rate after the second fentanyl dose 6 hours later, and Figure 4e The ventilation rate per minute was calculated after the third fentanyl dose, 24 hours later. Animals were monitored in a plethysmography chamber 90 minutes after each fentanyl dose. Figure 4fMale Sporgodory rats were administered 60 µmol / kg -1 The test compound (equivalent to 20 mg kg) -1 Naloxone and 63 mg kg -1 A single intravenous (iv) injection of AG10-L-E2-naloxone (per group) n = 3 The concentrations of the test compound in plasma were determined at different time points and expressed as the mean ± sd of three biological replicates. Figures 4g to 4i Reversal of fentanyl-induced OIRD by intravenous (iv) administration of naloxone and AG10-L-E2-naloxone. Figure 4g The first dose of fentanyl (3 mg kg) was plotted. -1 (intravenous dose) and the antagonist 1 min later (60 µmol kg) -1 Minute ventilation over time (mean ± sd) following a single intravenous dose; per group n = 6 (sub-biological repetition). Figure 4h For the second dose of fentanyl (1.5 mg kg) -1 The minute ventilation rate following the intravenous dose. Figure 4i This is the third dose of fentanyl (1.5 mg kg). -1 Ventilation rate per minute following the intravenous dose. Monitor animals in a plethysmography chamber 90 minutes after each fentanyl dose. (See above...) Figures 3f to 3i The volumetric data described in the text are analyzed.

[0042] Figure 5 shows the preclinical evaluation and pharmacokinetic studies of AG10-L-E2-naloxone in monkeys. Figures 5a to 5d The efficacy of a mixture of AG10-L-E2-naloxone and low-dose free naloxone in fentanyl-induced OIRD in rats was evaluated. AG10-L-E2-naloxone (60 µmol kg -1 Equivalent to 63 mg kg -1 ) and naloxone (3 µmol kg -1 Equivalent to 1 mg / kg -1 The combination of single subcutaneous doses of fentanyl yielded excellent efficacy data, with complete reversal of OIRD after all three fentanyl doses. Minute ventilation (mean ± sd; per group) n = 6 (sub-biological repetition). Figures 5e to 5g To evaluate the potential of using AG10-L-E2-naloxone as a prophylactic agent against fentanyl in rats. Initially, AG10-L-E2-naloxone (60 µmol kg) was used. -1 ) or naloxone (15 µmol kg-1 and 60 µmol kg -1 The rats were administered the drug. Six hours later, a single dose of fentanyl (1.5 mg / kg) was given. -1 Rats were attacked with a subcutaneous dose. Minute ventilation (mean ± SD) per group n = 6 (Secondary biological repetition). As mentioned above... Figures 3f to 3i The volumetric data described in the text are analyzed. Figure 5h AG10-L-E2-naloxone (60 µmol kg) -1 Effects of subcutaneous dose (60 µmol / kg) on ​​rat body weight over four weeks. Three groups of rats were used. The first group was treated with the medium. The second group was treated with AG10-L-E2-naloxone (60 µmol / kg). -1 The third group was treated with fentanyl, followed by AG10-L-E2-naloxone (60 µmol kg). -1 Treatment was initiated with fentanyl at 6 hr and 24 hr. Body weight is expressed as mean ± SD (per group). n = 6 (sub-biological repetition). Figures 5i to 5k After 72 hours, the cytotoxicity of AG10-L-E2-naloxone and its metabolite AG10-L-E2-COOH (100 μM) to various cell lines was assessed using an MTT assay. Doxorubicin (10 μM) was used as a positive control. Figure 5i , HeLa; cervical cancer cell lineage. Figure 5j Hep3B; human liver tumor cell line. Figure 5k MCF7; breast cancer cell line. Each bar shows the mean ± sd ( n = 4 (repeated once). Figure 5l Three male crab-eating macaques were administered a single subcutaneous (sc) injection of AG10-L-E2-naloxone (30 µmol kg). -1 Equivalent to 31 mg kg -1 The concentrations of AG10-L-E2-naloxone and naloxone released from AG10-L-E2-naloxone in the plasma of each monkey were measured at different time points and expressed as the mean ± sd of three biological replicates.

[0043] Figure 6 Display AG10-L-E2-naloxone ( Figure 6 a) AG10-L-E2-COOH ( Figure 6 b) and AG10-L-E2-ethyl ( Figure 6c) Calculated pKa values ​​and their predicted ionization in water (acidic, pH 3.5, obtained by dissolving HCl salts of these conjugates in water) and at physiological pH (7.4).

[0044] Figure 7 This study demonstrates the docking of AG10-L-E2-naloxone and known CES2-selective prodrug esters with CES2 and CES1. Mouse CES2 enzyme (pdb id: 8AXC) 1 The catalytic triplet residues S230, E347, and H459 are shown as black rods. Magnification highlights show a distance of 2.9 Å between the nucleophile S230 and the carbonyl ester of AG10-L-E2-naloxone. The distances between S230 and the carbonyl esters of the prodrugs irinotecan, procaine, and molnupiravir are 3.3 Å, 3.1 Å, and 3.5 Å, respectively. The prodrugs and CES1 (pdb id: 1MX9) are also shown. 2 Docking studies showed that irinotecan, procaine, and monopravir had longer distances, at 9.2 Å, 6.8 Å, and 4.8 Å, respectively. Docking studies also showed that AG10-L-E2-naloxone did not bind to CES1.

[0045] Figure 8 shows the chemical structure and stability of the newly synthesized aryl-based naloxone prodrug. Figure 8a The chemical structures of newly synthesized aryl-based naloxone prodrugs BA naloxone, Boc-PABA-naloxone, PABA-naloxone, and known CES2 selective prodrugs are described. Figure 8b and Figure 8c To assess the stability of the prodrug in human plasma (tested at 100 μM), and Figures 8d to 8i Stability in rat plasma was assessed. The stability of the compound (tested at 100 μM) in rat plasma was obtained in the absence and presence of the CES2 inhibitor loperamide and the CES1 inhibitor digitoxin. Known CES2-selective substrates procaine (local anesthetic), monopravir (therapeutic for COVID-19), and irinotecan (anticancer drug) were included for comparison. The acyl moiety of the ester is shown in red. The concentration remaining in individual culture media was determined by HPLC at specified time points. Bar graphs represent the mean ± SD of the percentage of remaining compound. n = 3 (Repeated individually). Two-way ANOVA (mixed model) was used, followed by Tukey post-hoc test to determine statistical differences.

[0046] Figure 9 To demonstrate the effect of TTR on the in vitro pharmacokinetics of AG10-L-E2-naloxone. Figure 9a. Stability of AG10-L-E2-naloxone (tested at 5 μM) in rat plasma in the absence and presence of the selective TTR ligand AG10 (10 μM). Figure 9 b. Metabolic bioactivation of AG10-L-E2-naloxone (5 μM) in human liver microsomes (HLM) in the presence and absence of human TTR (5 μM). Figure 9 c. The effect of AG10-L-E2-naloxone (5 μM) on the binding of human TTR (5 μM) in buffer was evaluated using rapid equilibration dialysis with a molecular weight cutoff (MWCO) of 8 kDa. Figure 9 a to Figure 9 The bar chart for c represents the average of the remaining / bound compound percentage ± sd ( n = 3 (repeated once). For Figure 9 a, Figure 9 b. Using a two-way ANOVA (mixed model), followed by the Holm-Šídák test to determine statistical differences, and for Figure 9 c. One-way ANOVA was used, followed by Tukey's post-hoc test to determine statistical differences. For Figure 9 a, F(2,8) = 62.29, (compared to AG10-L-E2-naloxone, * P <0.0001). For Figure 9 b, F(1,4) = 223.4, (compared to AG10-L-E2-naloxone, * P <0.0001). For Figure 9 c, F(2,6) = 3216, (compared to AG10-L-E2-naloxone or AG10 without TTR, * P <0.0001).

[0047] Figure 10 Evaluation of blood-brain barrier penetration of AG10-L-E2-naloxone and AG10-L-E2-COOH. Figure 10 a, the brain concentration to plasma concentration ratio of AG10-L-E2-naloxone, AG10-L-E2-COOH, naloxone derived from AG10-L-E2-naloxone, and naloxone administered in free form, and Figure 10 b, CSF to plasma concentration ratio. This was determined by a single subcutaneous injection of 15 µmol / kg. -1 The test compound (equivalent to 5 mg kg) -1 Naloxone and 15.8 mg / kg -1 AG10-L-E2-naloxone) in male Sporgodory rats (each group) n = 3Dosage administration. Bar graphs show the individual means ± sd for three biological replicates. Two-way ANOVA was used, followed by the Holm-Šídák test to determine statistical differences (compared to naloxone, * P <0.0001). For the brain-plasma ratio experiment, F(1,4) = 59.34. P <0.0001, and for the CSF plasma ratio experiment, F(1,4) = 35.87, P <0.0001.

[0048] Figure 11 Presents pharmacodynamic evaluations of repeated doses of AG10-L-E2-naloxone in rats. Includes fentanyl overdose mortality data and rescue effects of multiple low doses of naloxone on male Sporgodory rats (per group). n = 8 biological replicates). Fentanyl dosing regimen (subcutaneous dose of 3 mg / kg at 0 hr). -1 and 3 mg kg at 6 hr -1 Group 1 (red line): Administration via a carrier. Group 2 (gray line): 5 min after the first fentanyl dose, first administered a single subcutaneous dose of 3 µmol / kg. -1 Naloxone (equivalent to 1 mg kg) -1 ) was administered, followed by five smaller doses of naloxone (0.6 µmol / kg per dose) at 1.5, 2.5, 3.5, 4.5, and 5.5 hours. -1 Naloxone; equivalent to 0.2 mg / kg per dose. -1 ) Administration. Group 3 (blue line): 5 min after the first fentanyl dose, administer a single subcutaneous dose of 3 µmol / kg. -1 Naloxone (equivalent to 1 mg / kg) -1 ) was administered, followed by ten smaller doses of naloxone (0.6 µmol / kg per dose) at 1.5, 2.5, 3.5, 4.5, 5.5, 6.5, 7.5, 8.5, 9.5, and 10.5 hours. -1 Naloxone; equivalent to 0.2 mg / kg per dose. -1 Dosage was administered. Statistical differences were determined using the Mantel-Cox test followed by multiple comparisons with Bonferroni correction (α = 0.016). Compared to the medium, * P <0.0001, and compared to the medium, ** P <0.0026.

[0049] Figure 12This study presents a comparison of the pharmacokinetic profiles of low-dose and high-dose subcutaneous administration of naloxone over a period of up to 6 hours. Naloxone pharmacokinetic profiles (data from...) Figure 3a and Figure 5a ) shows with 5 mg kg -1 The difference in dosage is likely due to temporary saturation of the clearance pathway, with higher naloxone doses (20 mg / kg) being more likely to be attributed to this. -1 The sc) showed slower clearance from 0.5 hr to 2 hr. After 2 hr, both doses exhibited similar clearance rates and linear kinetics, indicating that tissue redistribution was limited at higher doses.

[0050] Figure 13 Demonstrates the reversal of intravenous (iv) administration of fentanyl OIRD in male Spögdolly rats with multiple low doses of naloxone or AG10-L-E2-naloxone. Figure 13 a. Dosage protocol for fentanyl and its antagonists. At the first fentanyl dose (3 mg / kg) -1 Administer naloxone (0.6 µmol kg / intravenous dose) over 1 minute. -1 Equivalent to 0.2 mg kg -1 Then, five small intravenous doses were administered at 2, 3, 4, 5, and 6 hours, followed by a second dose of fentanyl (1.5 mg kg) at 6 hours. -1 (Intravenous dose) (per group) n = 6 At the first fentanyl dose (3 mg kg) -1 Administer AG10-L-E2-naloxone (15 µmol kg / intravenous dose) over 1 minute. -1 Equivalent to 5 mg kg -1 Then, administer a second dose of fentanyl (1.5 mg / kg) at 4 hours. A second dose of fentanyl (1.5 mg / kg) is administered at 6 hours. -1 (Intravenous dose). Reversal of OIRD is represented by minute ventilation relative to baseline (minute ventilation in rats prior to fentanyl injection is considered baseline). Figure 13 b plots minute ventilation (mean ± sd) over time after the first dose of fentanyl and the antagonist; per group n = 6 (Secondary biological repetition) Figure 13c represents minute ventilation 6 hours after the second fentanyl dose. Animals were monitored in a plethysmography chamber 90 minutes after each fentanyl dose. Data were analyzed using GraphPad Prism V9.3.1.471 software (La Jolla, CA, USA). Prior analysis revealed that a statistical power of 0.8 was required for our experimental SD value, assuming an α of 0.05. Power analysis was performed using G*Power software (3.0.10). Statistical differences were determined using two-way ANOVA with repeated measures followed by Tukey's post-hoc test. A significant difference was determined at (#) P < 0.05.

[0051] Figures 14a to 14e The results of rat plasma stability assays for AG10-L-E2-naloxone, AG10-L-PHBA-E-naloxone, PEG-linker-E2-naloxone, AG10-short-L-E2-naloxone, and AG10-L-m-amino-E-naloxone are presented respectively.

[0052] Figure 15 Results of human plasma stability assays for AG10-L-E2-naloxone, AG10-L-PHBA-E-naloxone, PEG-linker-E2-naloxone, AG10-short-L-E2-naloxone, and AG10-L-m-amino-E-naloxone are presented.

[0053] Figures 16a to 16f The results of rat plasma stability assays for AG10-L-E2-acetaminophen, AG10-L-E2-propofol, AG10-L-E2-estrone, AG10-L-E2-endorphin-2, AG10-L-E2-Tyr-Ala, and AG10-L-E2-eporphin are presented respectively.

[0054] Figure 17 Results of human plasma stability assays for AG10-L-E2-acetaminophen, AG10-L-E2-propofol, AG10-L-E2-estrone, AG10-L-E2-endorphin-2, AG10-L-E2-Tyr-Ala, and AG10-L-E2-eporphin are presented. Detailed Implementation

[0055] Here, we develop a novel prodrug approach for enhancing the DOA of drugs without affecting their biological activity. Our approach involves conjugating a drug to a selective TTR ligand via an ester moiety. The approach described herein can be used for a variety of drug moieties, including small molecules, peptides, and proteins. The use of a selective TTR ligand, as illustrated herein, utilizes endogenous TTRs, representing a major advantage of this novel prodrug approach. TTRs are 55 kDa homotetrameric proteins primarily secreted from the liver into the bloodstream with a circulating half-life of approximately 2 days. The primary function of TTRs in humans (plasma concentration of approximately 5 μM) is to transport holo-retinol-binding proteins (RBPs bound to retinol or vitamin A) in the blood. Using orthogonal sites to holo-retinol, TTRs act as a backup carrier for thyroxines (<1% thyroxine binding).

[0056] Our approach possesses several unique features that will make it ideal for controlling drug release and prolonging drug DOA. First, the prodrug can bind to the TTR, thereby extending its DOA to 2 days or longer. Second, the prodrug has balanced hydrophilicity, which is crucial for rapid subcutaneous absorption to provide sufficient concentrations of antidote for effective rescue. Third, the balanced hydrophilicity, small size, and ease of synthesis of our conjugate (we do not use polymers) will make it ideal for subcutaneous (sc) administration, offering the advantage of ease of administration while simultaneously providing a prolonged prodrug duration compared to intravenous routes. Fourth, the modular approach will allow us to produce prodrugs at adjustable hydrolysis rates, which can be used for rescue, protection (prophylaxis), or both.

[0057] definition "Individual" or "subject" refers to a mammal and includes both human and non-human mammals. Examples of individuals include, but are not limited to, mice, rats, hamsters, guinea pigs, rabbits, cats, dogs, goats, sheep, cows, monkeys, and humans. In some implementations, an individual or subject refers to a human.

[0058] As used herein, the term "about" includes and describes the parameter or value itself. For example, "about X" includes and describes X itself.

[0059] "Treatment" or "treating" refers to a means of achieving a beneficial or desired outcome, including clinical outcomes. Beneficial or desired outcomes may include one or more of the following: reducing one or more symptoms caused by a disease or condition; weakening the severity of a disease or condition; slowing or halting the development of one or more symptoms associated with a disease or condition (e.g., stabilizing the disease or condition, preventing or delaying the worsening or progression of the disease or condition); and alleviating the disease, for example by eliminating clinical symptoms (e.g., improving the disease state, enhancing the effect of another medication, delaying the progression of the disease, improving quality of life, and / or prolonging survival).

[0060] As used herein, “delaying” the development of a disease or condition means postponing, hindering, slowing, delaying, stabilizing, and / or slowing the progression of a disease or condition. This delay can vary in length depending on the patient’s medical history and / or the individual being treated. As will be apparent to those skilled in the art, sufficient or significant delay can effectively encompass prevention, ensuring that the individual does not develop a disease or condition.

[0061] As used herein, the terms "therapeutic effective amount" or "effective amount" mean the amount of a compound of this disclosure or its pharmaceutically acceptable salt that is sufficient to achieve a therapeutic effect when administered to an individual. As understood in the art, an effective amount can be a single or multiple doses, for example, a single or multiple doses may be required to achieve the desired therapeutic endpoint. An effective amount can be considered in the context of administration of one or more therapeutic agents, and a single agent may be considered to be administered at an effective amount if the desired or beneficial outcome can be achieved or has been achieved in combination with one or more other agents.

[0062] As used herein, “unit dosage form” refers to a compound suitable as a unit dose, each unit containing a predetermined amount of active ingredient or compound in a pharmaceutically acceptable carrier.

[0063] As used herein, “pharmaceutically acceptable” means a material that is biologically or otherwise desirable, for example, that can be incorporated into a pharmaceutical composition administered to an individual without causing significant undesirable biological effects.

[0064] As used herein, the term "half-life" may be used to refer to the loss of half of its activity (e.g., its pharmacological, physiological, or radiological activity). The term "half-life" may be used interchangeably with the terms "biological half-life" or "terminal half-life." In some embodiments, the half-life of a substance is the time it takes for a substance (e.g., a metabolite, drug, signaling molecule, radionuclide, or other substance) to lose half of its pharmacological, physiological, or radiological activity. In some embodiments, the term "half-life" may be used to refer to bodily purification, which eliminates a substance from the body via renal and hepatic function plus excretion. In some embodiments, the term "half-life" may be used to refer to the time it takes for the plasma concentration of a substance to halve to its steady-state level (plasma half-life).

[0065] As used herein, the terms “joint,” “link,” and “linking group” refer to a chemical linking portion that connects two groups and has a main chain length of 50 atoms or less. In some embodiments, the joint or link may be a covalent bond connecting two groups, or a chemical portion having a chain with a length ranging from 1 to 50 atoms, 2 to 20 atoms, 3 to 30 atoms, 4 to 40 atoms, 10 to 50 atoms, 10 to 30 atoms, 12 to 26 atoms, 14 to 30 atoms, or increasing in length by an amount of 1 atom. For example, a joint or a portion thereof may have a length of 1, 2, 3, 4, 5, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 45, or 50 carbon atoms, wherein the joint may be a straight chain, branched chain, cyclic chain, or a single atom. In some embodiments, the length of the joint may be 10 angstroms, 11 angstroms, 12 angstroms, 13 angstroms, 14 angstroms, 15 angstroms, 16 angstroms, 17 angstroms, 18 angstroms, 19 angstroms, 20 angstroms, 22 angstroms, 23 angstroms, 24 angstroms, 25 angstroms, 26 angstroms, 27 angstroms, 28 angstroms, 29 angstroms, 30 angstroms, 32 angstroms, 34 angstroms, 36 angstroms, 38 angstroms, 40 angstroms, 45 angstroms, 50 angstroms, or any amount in increments of 1 angstrom. In some embodiments, one, two, three, four, or five or more carbon atoms in the linker backbone may optionally be substituted with sulfur, nitrogen, or oxygen heteroatoms. The bonds between the backbone atoms may be saturated or unsaturated, typically with no more than one, two, or three unsaturated bonds present in the linker backbone. The linker may include one or more substituents, such as alkyl, aryl, or alkenyl groups. The linker may include, but is not limited to, oligomers (ethylene glycol); ethers; thioethers; tertiary amines; and may be straight-chain or branched alkyl groups, such as methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1,1-dimethylethyl (tert-butyl), etc. The linker backbone may include cyclic groups, such as aryl, heterocyclic, or cycloalkyl groups, wherein the backbone includes two or more atoms of the cyclic group, such as two, three, or four atoms. The linker may be cleavable or non-cleavable.

[0066] The linker can use any suitable functional group (formic acid, amine, alcohol, carbamate, ester, amide, ether, thioether, maleic anhydride, etc.) and linker chemicals conjugated with TTR ligands and Y. For example, the conjugation chemistry described by GT Hermanson (“Bioconjugate Techniques”, Academic Press, 2nd ed., 2008) is readily applicable to the preparation of the subject heterobifunctional compounds and is hereby incorporated herein by reference in its entirety.

[0067] As used herein, the term "alkyl" refers to a non-branched or branched saturated monovalent hydrocarbon chain. As used herein, alkyl groups have 1–20 carbon atoms (i.e., C64-C64). 1-20 Alkyl groups), 1-16 carbons (i.e., C16) 1-16 Alkyl groups), 1-12 carbons (i.e., C46) 1-12 Alkyl groups), 1-10 carbons (i.e., C45) 1-10 Alkyl groups), 1-8 carbons (i.e., C64) 1-8 Alkyl groups), 1-6 carbons (i.e., C64) 1-6 Alkyl groups), 1-4 carbons (i.e., C4) 1-4 Alkyl groups or 1-3 carbons (i.e., C14-C24) 1-3 Alkyl groups. Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, pentyl, 2-pentyl, isopentyl, neopentyl, hexyl, 2-hexyl, 3-hexyl, and 3-methylpentyl. When an alkyl residue having a specific number of carbon atoms is named by its chemical name or molecular formula, it may encompass all positional isomers having that number of carbon atoms; for example, "butyl" includes n-butyl, sec-butyl, isobutyl, and tert-butyl; and "propyl" includes n-propyl and isopropyl. Certain commonly used alternative names may be used, and will be understood by those skilled in the art. For example, a divalent group, such as a divalent "alkyl", may be referred to as an "alkylene".

[0068] As used herein, the term "alkenyl" refers to a branched or unbranched monovalent hydrocarbon chain containing at least one carbon-carbon double bond. As used herein, alkenyl groups have 2–20 carbons (i.e., C60-C60). 2-20 alkenyl), 2-16 carbons (i.e., C) 2-16 alkenyl), 2-12 carbons (i.e., C) 2-12 alkenyl), 2-10 carbons (i.e., C10) 2-10 alkenyl), 2-8 carbons (i.e., C) 2-8 alkenyl), 2-6 carbons (i.e., C10) 2-6 alkenyl), 2-4 carbons (i.e., C10) 2-4 Alkenyl group or 2-3 carbons (i.e., C14-C24) 2-3 Alkenyl groups. Examples of alkenyl groups include, but are not limited to, vinyl, 1-propenyl, 2-propenyl, 1,2-butadienyl, and 1,3-butadienyl. When an alkenyl residue having a specific number of carbon atoms is named by its chemical name or molecular formula, it may encompass all positional isomers having that number of carbon atoms; for example, "propenyl" includes both 1-propenyl and 2-propenyl. Certain commonly used alternative names may be used, and will be understood by those skilled in the art. For example, a divalent group, such as a divalent "alkenyl," may be referred to as an "alkenylene."

[0069] As used herein, the term "alkynyl" refers to a branched or unbranched monovalent hydrocarbon chain containing at least one carbon-carbon triple bond. As used herein, an alkynyl group has 2–20 carbons (i.e., C60-C60). 2-20alkynyl group), 2-16 carbons (i.e., C16) 2-16 alkynyl group), 2-12 carbons (i.e., C12) 2-12 alkynyl group), 2-10 carbons (i.e., C10) 2-10 alkynyl group), 2-8 carbons (i.e., C14-C24) 2-8 alkynyl group), 2-6 carbons (i.e., C64) 2-6 alkynyl group), 2-4 carbons (i.e., C14) 2-4 (alkynyl group) or 2-3 carbons (i.e., C10) 2-3 (Alynyl). Examples of alkynyl groups include, but are not limited to, ethynyl, propyn-1-alkynyl, propyn-2-alkynyl, butyn-1-alkynyl, butyn-2-alkynyl, and butyn-3-alkynyl. When an alkynyl residue having a specific number of carbon atoms is named by its chemical name or molecular formula, it may encompass all positional isomers having that number of carbon atoms; for example, "propynyl" includes both propyn-1-alkynyl and propyn-2-alkynyl. Certain commonly used alternative names may be used, and will be understood by those skilled in the art. For example, a divalent group, such as a divalent "alkynyl," may be referred to as "acetylenyl."

[0070] As used herein, the term "alkoxy" refers to the -O-alkyl moiety. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy, n-hexyloxy, and 1,2-dimethylbutoxy.

[0071] As used herein, the term "aryl" refers to a fully unsaturated carbocyclic moiety. The term "aryl" encompasses both monocyclic and polycyclic fused-ring moieties. As used herein, aryl encompasses moieties containing, for example, 6 to 20 ring carbon atoms (i.e., C24 carbon atoms). 6-20 aryl), 6 to 16 ring carbon atoms (i.e., C 6-16 aryl), 6 to 12 ring carbon atoms (i.e., C 6-12 aryl group or 6 to 10 cyclic carbon atoms (i.e., C46) 6-10 The ring moiety of an aryl group. Examples of aryl moiety include, but are not limited to, phenyl, naphthyl, fluorenyl, and anthracene.

[0072] As used herein, the term "cycloalkyl" refers to a saturated or partially unsaturated carbocyclic moiety. The term "cycloalkyl" encompasses both monocyclic and polycyclic cyclic moieties, wherein the polycyclic moiety may be a fused ring, branched ring, or spirocyclic ring. Cycloalkyl includes cycloalkenyl groups, wherein the cyclic moiety contains at least one cyclic double bond. Cycloalkyl includes any polycyclic carbocyclic moiety containing at least one non-aromatic ring, regardless of its connection point to the rest of the molecule. As used herein, cycloalkyl includes moieties containing, for example, 3 to 20 cyclic carbon atoms (i.e., C14 carbon atoms). 3-20 cycloalkyl groups, 3 to 16 cyclic carbon atoms (i.e., C1646) 3-16 cycloalkyl groups), 3 to 12 cyclic carbon atoms (i.e., C12) 3-12 cycloalkyl groups), 3 to 10 cyclic carbon atoms (i.e., C145-C ... 3-10 cycloalkyl groups), 3 to 8 cyclic carbon atoms (i.e., C14 and C24)3-8 cycloalkyl groups), 3 to 6 cyclic carbon atoms (i.e., C1646) 3-6 cycloalkyl groups or 3 to 5 cyclic carbon atoms (i.e., C14-C5 ... 3-5 The cycloalkyl ring. Monocyclic cycloalkyl ring moiety includes, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic groups include, for example, bicyclic [2.2.1]heptyl, bicyclic [2.2.2]octyl, adamantyl, norcamphenyl, decahydronaphthyl, 7,7-dimethyl-bicyclic [2.2.1]heptyl, etc. In addition, cycloalkyl also includes spirocycloalkyl ring moiety, such as spiro[2.5]octyl, spiro[4.5]decyl, or spiro[5.5]undecyl.

[0073] As used herein, the term "halogen" refers to an atom occupying Group VIIA of the periodic table and includes fluoro, chloro, bromo, and iodo. Additionally, terms such as "halogenated alkyl" are intended to include both monohalogenated and polyhalogenated alkyl groups. For example, the term "C"... 1-4 "Halogenated alkyl" is intended to include trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, difluoromethyl, etc.

[0074] As used herein, the term "heteroaryl" refers to an aromatic (fully unsaturated) ring moiety containing one or more cyclic heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur. The term "heteroaryl" includes both monocyclic and polycyclic fused-ring moieties. As used herein, a heteroaryl contains, for example, 5 to 20 ring atoms (i.e., 5- to 20-membered heteroaryl), 5 to 16 ring atoms (i.e., 5- to 16-membered heteroaryl), 5 to 12 ring atoms (i.e., 5- to 12-membered heteroaryl), 5 to 10 ring atoms (i.e., 5- to 10-membered heteroaryl), 5 to 8 ring atoms (i.e., 5- to 8-membered heteroaryl), or 5 to 6 ring atoms (i.e., 5- to 6-membered heteroaryl). Any monocyclic or polycyclic aromatic ring moiety containing one or more cyclic heteroatoms is considered a heteroaryl, regardless of its connection point to the rest of the molecule (i.e., the heteroaryl moiety may be connected to the rest of the molecule via any cyclic carbon or any cyclic heteroatom of the heteroaryl moiety). Examples of heteroaryl groups include, but are not limited to, oxazolyl, thiazolyl, pyridyl, pyrimidinyl, and pyridazinyl.

[0075] As used herein, the term "heterocyclic group" refers to a saturated or partially unsaturated cyclic moiety encompassing one or more cyclic heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur. The term "heterocyclic group" includes both monocyclic and polycyclic cyclic moieties, wherein polycyclic cyclic moieties can be fused rings, bridged rings, or spirocyclic rings. Any non-aromatic monocyclic or polycyclic cyclic moieties containing at least one cyclic heteroatom are considered heterocyclic groups, regardless of their connection points to the remainder of the molecule (i.e., the heterocyclic group moieties may be connected to the remainder of the molecule via any cyclic carbon or any cyclic heteroatom of the heterocyclic group moieties). Furthermore, the term "heterocyclic group" is intended to encompass any polycyclic cyclic moieties containing at least one cyclic heteroatom, wherein the polycyclic cyclic moieties contain at least one non-aromatic ring, regardless of their connection points to the remainder of the molecule. As used herein, heterocyclic groups comprise, for example, 3 to 20 ring atoms (i.e., 3- to 20-membered heterocyclic groups), 3 to 16 ring atoms (i.e., 3- to 16-membered heterocyclic groups), 3 to 12 ring atoms (i.e., 3- to 12-membered heterocyclic groups), 3 to 10 ring atoms (i.e., 3- to 10-membered heterocyclic groups), 3 to 8 ring atoms (i.e., 3- to 8-membered heterocyclic groups), 3 to 6 ring atoms (i.e., 3- to 6-membered heterocyclic groups), 3 to 5 ring atoms (i.e., 3- to 5-membered heterocyclic groups), 5 to 8 ring atoms (i.e., 5- to 8-membered heterocyclic groups), or 5 to 6 ring atoms (i.e., 5- to 6-membered heterocyclic groups). Examples of heterocyclic groups include, for example, aza-butyl, piperidinyl, piperazine, pyrrolyl, pyrazolyl, thiazolyl, tetrahydrofuranyl, or tetrahydropyranyl. Examples of spiroheterocyclic rings include, but are not limited to, bicyclic and tricyclic ring systems, such as oxabicyclo[2.2.2]octyl, 2-oxa-7-azaspiro[3.5]nonyl, 2-oxa-6-azaspiro[3.4]octyl, and 6-oxa-1-azaspiro[3.3]heptyl. Examples of fused heterocyclic rings include, but are not limited to, 5,6,7,8-tetrahydropyridino[4,3-d]pyrimidinyl, 6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidinyl, 5,7-dihydrofurano[3,4-d]pyrimidinyl, and 5,7-dihydrothieno[3,4-d]pyrimidinyl, wherein the heterocyclic group may be linked via any ring of the fused system.

[0076] As used in this article, the term "oxo group" refers to the =O part.

[0077] As used herein, the terms “optional” and “optionally” mean that the event or situation described below may or may not occur, and the description includes instances where the event or situation occurs and instances where it does not occur. Thus, the term “optionally substituted” means that any one or more (e.g., 1, 2, 1 to 5, 1 to 3, 1 to 2, etc.) hydrogen atoms on a specified atom or part or group may be substituted or not substituted. By way of illustration and not limitation, the phrase “methyl group optionally substituted with one or more chlorine atoms” encompasses the -CH3, -CH2Cl, -CHCl2, and -CCl3 moieties.

[0078] It should be understood that aspects and implementation schemes described herein as “comprising” include “consisting of the implementation scheme” and “substantially consisting of the implementation scheme”.

[0079] As used herein, the term "pharmaceutically acceptable salt" for a given compound refers to a salt that retains the biological efficacy and properties of the given compound and is biologically or otherwise desirable. "Pharmaceutically acceptable salt" includes, for example, salts formed with inorganic acids and salts formed with organic acids. Furthermore, if the compound described herein is obtained as an acid addition salt, the free base can be obtained by alkalizing the acid salt solution. Conversely, if the product is a free base, the addition salt, especially a pharmaceutically acceptable addition salt, can be produced according to the conventional procedure for preparing acid addition salts from base compounds by dissolving the free base in a suitable organic solvent and treating the solution with acid. See, for example... Handbook of Pharmaceutical Salts Properties, Selection, and Use [Reference to John Wiley & Sons (2008), International Union of Pure and Applied Chemistry, which is incorporated herein by reference in its entirety.] Those skilled in the art will recognize the various synthetic methods that can be used to prepare pharmaceutically acceptable addition salts that are non-toxic. Pharmaceutically acceptable acid addition salts can be prepared from inorganic or organic acids. Salts derived from inorganic acids include, for example, salts of hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc. Salts derived from organic acids include, for example, salts of acetic acid, propionic acid, gluconic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, trifluoroacetic acid, etc. Similarly, pharmaceutically acceptable base addition salts can be prepared from inorganic or organic bases. Salts derived from inorganic bases include sodium, potassium, lithium, aluminum, ammonium, calcium, and magnesium salts, for example only. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines. Specific examples of suitable amines, for example only, include isopropylamine, trimethylamine, diethylamine, tris(isopropyl)amine, tris(n-propyl)amine, ethanolamine, 2-dimethylaminoethanol, piperazine, piperidine, morpholine, N-ethylpiperidine, etc.

[0080] Some of the compounds described herein can exist in tautomer form. The tautomers are in equilibrium with each other. For example, amide-containing compounds can exist in equilibrium with imine tautomers. Regardless of the tautomers shown and regardless of the equilibrium nature between the tautomers, those skilled in the art will understand that the compounds disclosed herein comprise both amide and imine tautomers. Therefore, for example, amide-containing compounds should be understood to include their imine tautomers. Similarly, imine-containing compounds should be understood to include their amide tautomers.

[0081] The compounds disclosed herein, or pharmaceutically acceptable salts thereof, may include asymmetric centers and thus may produce enantiomers, diastereomers, and other stereoisomers, which may be defined in absolute stereochemistry as ( R )-or( S (R)- and (S)-, or (D)- and (L)- for amino acids. This disclosure is intended to include all such possible isomers, as well as their racemic and optically pure forms and mixtures thereof in any ratio. Optically active (+) and (-), (R)- and (S)-, or (D)- and (L)- isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques such as chromatography and / or stepwise crystallization. Conventional techniques for preparing / separating individual enantiomers include chiral synthesis from suitable optically pure precursors or resolution of racemic products (or racemic products of salts or derivatives) using, for example, chiral high-performance liquid chromatography (HPLC) and chiral supercritical fluid chromatography (SFC). When the compounds described herein contain alkene double bonds or other geometrically asymmetric centers, this disclosure is intended to include both E and Z geometric isomers unless otherwise specified. Similarly, cis and trans are used in their conventional sense to describe relative spatial relationships.

[0082] Conjugate In one respect, this paper provides a conjugate of formula (I): Or its pharmaceutically acceptable salt, wherein: T is a ligand selective for transthyretin-TTR; L stands for connector; L B Click products that do not exist or are formed through a click reaction between the first click handle and the second click handle; E is a cleavable ester; and D 1 This refers to the pharmaceutical component.

[0083] In some embodiments, the connector (L) has a size of about 10 Å to about 40 Å. In some embodiments, the connector (L) has a size of about 10 Å to about 40 Å. In some embodiments, the connector (L) has a size of about 15 Å to about 30 Å. In some embodiments, the connector (L) has a size of about 20 Å to about 30 Å. In some embodiments, the connector (L) has a size of about 20 Å. In some embodiments, the connector (L) has a size of about 25 Å. In some embodiments, the connector (L) has a size of about 30 Å.

[0084] In some implementations, the connector (L) and the click product (L) B Both have a size of approximately 10 Å to approximately 50 Å. In some embodiments, the connector (L) and the click product (L) have a size of approximately 10 Å to approximately 50 Å. B Both have a size of approximately 15 Å to approximately 45 Å. In some embodiments, the connector (L) and the click product (L) have a size of approximately 15 Å to approximately 45 Å. B Together, they have a size of approximately 25 Å to approximately 40 Å. In some embodiments, the connector (L) and the click product (L) B Together, they have a size of approximately 20 Å. In some embodiments, the connector (L) and the click product (L) B Together, they have a size of approximately 25 Å. In some embodiments, the connector (L) and the click product (L) B Together, they have a size of approximately 30 Å. In some embodiments, the connector (L) and the click product (L) B Together, they have a size of approximately 35 Å. In some embodiments, the connector (L) and the click product (L) B They together have a size of approximately 40 Å.

[0085] In some implementations, L B The following are the click products: (i) Copper-catalyzed reactions between azides and alkynes; (ii) The reaction between azides and dibenzocyclooctene (DBCO); (iii) Inverse electron demand Diels-alder cycloaddition (IEDDA) between the anti-cyclooctene (TCO) moiety and the tetrazine ring, or (iv) Staudinger reaction between azides and phosphine.

[0086] In some implementations, the product L is used to form the click product L. B The first and second clickers can be any suitable click chemical pair (e.g., azide-DBCO, azide-BCN, Tz-TCO, meTz-TCO, etc.). In some embodiments, the click product can be formed using a copper-catalyzed click reaction. One such copper-catalyzed click reaction is the Huisgen 1,3-dipolar cycloaddition (CuAAC) between an azide and an alkyne (see example). people, J. Org. Chem. 2002, 67(9), 3057-3064 and Rostovtsev et al., Angew. Chem. Int. Ed.2002, 41 (14), 2596-2599). In some embodiments, the click product is a triazole moiety. In some embodiments, the click product can be formed using a bisorthogonal chemistry method.

[0087] In some implementations, click product L B It can be formed by any suitable electron-demanding Diels-Alder reaction. In some embodiments, the click product can be formed by an electron-demanding Diels-Alder reaction between the anti-cyclooctene (TCO) moiety on the first or second click shank and the tetrazine (Tz) ring on the first or second click shank (see, for example, Selvaraj et al.). Curr. Opin. Chem. Biol. 2013, 17 (5), 753-760 and Rossin et al., Bioconjugate Chem (2013, 24(7), 1210-1217). In some embodiments, the first click handle comprises a tetrazine ring and the second click handle comprises a TCO moiety. In some embodiments, the tetrazine ring is unsubstituted. In some such embodiments, the tetrazine ring is methyltetrazine. In some embodiments, the tetrazine ring is a 6-methyl-substituted tetrazine. In some embodiments, the click product is a dihydropyridazine moiety.

[0088] In some implementations, click product L B The triazine moiety. In some embodiments, the conjugate of formula (I) comprises one of the following moieties: , where X 2 -CH2-, -NR x -, -S-, or -O-, where R x As defined elsewhere herein. In some embodiments, the conjugate comprises a portion In some implementations, the conjugate comprises a portion .

[0089] In some implementations, L is C 4-20 alkylene, wherein the C 4-20 Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally selected from -O-, -NR. x The - and -S- groups are replaced, where L 1a Halogenated, C 1-6 Alkyl groups, or C groups substituted with halogen groups 1-6 Alkyl group, wherein R x H or C optionally substituted with a halogen group 1-6 Alkyl group. In some embodiments, L is C. 10-20 alkylene, wherein the C 10-20Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally replaced by groups selected from -O-, -NH-, -N(CH3)-, and -S-. In some embodiments, L is C 10-15 alkylene, wherein the C 10-15 Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally replaced by groups selected from -O-, -NH-, and -N(CH3)-. In some embodiments, L is C 5-10 alkylene, wherein the C 5-10 Alkylene is optionally surrounded by one or more L 1a The alkylene units are substituted, and one or more alkylene units are optionally replaced by groups selected from -O- and -NH-.

[0090] In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion .

[0091] In some implementation schemes, LL B Includes part In some implementation schemes, LL B Includes part In some implementations, LL B Includes part In some implementation schemes, LL B Includes part In some implementation schemes, LL B Includes part In some implementations, LL B Includes part In some implementation schemes, LLB Includes part .

[0092] In some implementation schemes, LL B Includes part or , where R 9 For H or C 1-4 Alkyl, R 10 For H or C 1-4 Alkyl group, and Q is -CH2-, -O-, -S-, -NH- or -N(C 1-4 alkyl)-.

[0093] In some embodiments, the formula (I) conjugate contains a portion , , , , , , , or ,in: Ring A is C 6-10 Aryl or 5- to 10-membered heteroaryl groups, among which The C 6-10 Aryl or 5- to 10-membered heteroaryl groups are optionally surrounded by one or more R groups. 7 Replace, where each R 7 Independently selected from the following groups: hydrogen, halogen atoms (e.g., Cl, Br, or F), hydroxyl group, -OCF3, -NH2, -NO2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 cycloalkyl and Indicator and connector (L or LL) B ( ) connection points.

[0094] In some embodiments, ring A is optionally substituted with a 5- to 10-membered heteroaryl group. Examples of heteroaryl groups include, but are not limited to, azacyclic butyl, pyrazolyl, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyrimidinyl, pyridazinyl, indolizyl, isoindolyl, indolyl, dihydroindolyl, indolyl, purine, quinazinyl, isoquinolinyl, quinolinyl, phthalazinyl, naphthylpyridyl, quinoxalinyl, quinazolinyl, terpineyl, pteridinyl, carbazole, carbolinyl, phenanthridine, etc. Acridine, phenanthroline, isothiazolyl, phenazinyl, isoxazolyl, phenoxazinyl, phenthiazinyl, imidazoalkyl, imidazoline, piperidinyl, piperazinyl, indololinyl, phthalimide, 1,2,3,4-tetrahydroisoquinolinyl, 4,5,6,7-tetrahydrobenzo[b]thiophene, thiazolyl, thiazoalkyl, thiophene, benzo[b]thiophene, morpholinyl, thiomorpholinyl, 1,1-dioxothiomorpholinyl, piperidinyl, pyrrolylyl, and tetrahydrofuranyl. In some embodiments, ring A is selected from the group consisting of: phenyl, pyridinyl, pyrimidinyl, oxazolyl, pyrazolyl, and thiazolyl, wherein the phenyl, pyridinyl, pyrimidinyl, oxazolyl, pyrazolyl, or thiazolyl is optionally substituted by one or more of the following: C 1-6 Alkyl, C 1-6 Alkoxy, hydroxy, halogen, -NH2, -NHC 1-4 amino, -N(C 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 3-6 cycloalkyl or -NO2, wherein the C 1-6 Alkyl or C 1-6 The alkoxy group is optionally substituted with one or more halogen groups. In some embodiments, ring A is selected from the group consisting of phenyl, pyridyl, and pyrimidinyl, wherein the phenyl, pyridyl, and pyrimidinyl groups are optionally substituted with one or more of the following: C 1-6 Alkyl, C 1-6 Alkoxy, hydroxy, halogen, -NH2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 3-6 cycloalkyl or nitro, wherein the C 1-6 Alkyl or C 1-6 The alkoxy group may optionally be substituted with one or more halogen groups.

[0095] In some embodiments, ring A is phenyl or a substituted phenyl group. In some such embodiments, ring A is substituted with a methyl, methoxy, halogen (e.g., Cl, Br, F or I), nitro, or CF3 group.

[0096] In some embodiments, the formula (I) conjugate contains a portion , , , , , or .

[0097] In some embodiments, the formula (I) conjugate contains a portion , or .

[0098] In some embodiments, the formula (I) conjugate contains a portion , , , , , , , or ,in: Each R 7 Independently selected from the following groups: hydrogen, halogen atoms (e.g., Cl, Br, or F), hydroxyl group, -OCF3, -NH2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 Cycloalkyl, p is 0, 1, 2 or 3, and Indicator and connector (L or LL) B ( ) connection points.

[0099] In some embodiments, the formula (I) conjugate contains a portion , , or , in: Ring A is C 6-10 Aryl or 5- to 10-membered heteroaryl groups, among which The C 6-10Aryl or 5- to 10-membered heteroaryl groups are optionally surrounded by one or more R groups. 7 Replace, where each R 7 Independently selected from the following groups: hydrogen, halogen atoms (e.g., Cl, Br, or F), hydroxyl group, -OCF3, -NH2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 cycloalkyl; and Indicates the connection point with the connector (L).

[0100] In some embodiments, the formula (I) conjugate contains a portion , , or , Each R 7 Independently selected from the following groups: hydrogen, halogen atoms (e.g., Cl, Br, or F), hydroxyl group, -OCF3, -NH2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 Cycloalkyl, p is 0, 1, 2 or 3, and Indicates the connection point with the connector (L).

[0101] In some embodiments, the formula (I) conjugate contains a portion , In some embodiments, the formula (I) conjugate contains a portion Each methylene group may optionally be substituted with: a halogen atom (e.g., Cl, Br, or F), a hydroxyl group, -OCF3, -NH2, or -NHC. 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy or C 3-6 Cycloalkyl.

[0102] In some embodiments, the formula (I) conjugate contains a portion , or , Each methylene group may optionally be substituted with: a halogen atom (e.g., Cl, Br, or F), a hydroxyl group, -OCF3, -NH2, or -NHC. 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy or C 3-6 cycloalkyl; And among them Indicator and connector (L or LL) B The connection point of ) where Indicates the connection point with the connector (L).

[0103] In some embodiments, the formula (I) conjugate contains a portion , or ,in Indicates the connection point with the connector (L).

[0104] In some implementation schemes, D 1 It is a peptide.

[0105] In some embodiments, the peptide is RGK-MCA. In some embodiments, the peptide is neurotensin. In some embodiments, the peptide is gonadotropin-releasing hormone (GnRH). In some embodiments, the peptide is GIP (1-42) human peptide. In some embodiments, the peptide is D6-GnRH. In some embodiments, the peptide is insulin. In some embodiments, the peptide is teriparatide. In some embodiments, the peptide is salmon calcitonin. In some embodiments, the peptide is iparene. In some embodiments, the peptide is vasopressin. In some embodiments, the peptide is glucagon. In some embodiments, the peptide is alexiside. In some embodiments, the peptide is goserelin. In some embodiments, the peptide is leuprolide. In some embodiments, the peptide is teriparatide. In some embodiments, the peptide is lixinaside. In some embodiments, the peptide is desmopressin. In some embodiments, the peptide is oxytocin. In some embodiments, the peptide is corticotropin. In some embodiments, the peptide is sermorelin. In some embodiments, the peptide is bumenotide. In some embodiments, the peptide is abalotropic peptide. In some embodiments, the peptide is dilorelin.

[0106] In some embodiments, the peptide drug is a glucagon-like peptide-1 (GLP-1) receptor agonist used to treat obesity and / or diabetes. In some such embodiments, the peptide drug is semaglutide, liraglutide, dulaglutide, abiglutide, or exenatide. Because the duration of action of the peptide drug is significantly prolonged when incorporated into the conjugates of this disclosure, GLP-1 agonist conjugates can reduce the frequency of administration, such as once weekly, once every two weeks, once every three weeks, or once monthly.

[0107] In some embodiments, the peptide drug is a glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic peptide (GIP) receptor agonist used to treat obesity and / or diabetes. In some such embodiments, the peptide drug is telposide. Because the duration of action of the peptide drug is significantly prolonged when incorporated into the conjugates of this disclosure, the GLP-1 / GIP agonist conjugates can reduce the frequency of administration, such as once weekly, once every two weeks, once every three weeks, or once monthly.

[0108] In some embodiments, the peptide is an endogenous opioid peptide. In some such embodiments, the peptide is endorphin-1 or endorphin-2. In other embodiments, the peptide is corticomerol.

[0109] In some implementation schemes, D 1 It is a protein.

[0110] In some implementation schemes, D 1The cytokine is used. In some embodiments, the cytokine is interleukin-2 (IL-2). In some embodiments, the cytokine is mutant IL-2, such as the cytokine described in US 9,206,943. In some embodiments, the cytokine is interleukin-15 (IL-15). In some embodiments, the cytokine is mutant IL-15. In one such embodiment, IL-15 is IL-15[N27D]. In some embodiments, the cytokine is interleukin-7 (IL-7). In some embodiments, the cytokine is interleukin-9 (IL-9). In some embodiments, the cytokine is interleukin-10 (IL-10). In some embodiments, the cytokine is interleukin-21 (IL-21). In some embodiments, the cytokine is interleukin-10 (IL-10).

[0111] In some implementation schemes, D 1 It is an anticancer agent. In some such implementations, D 1 For SN-38. In some such implementations, D 1 It is monomethylolpropionate E. In some such embodiments, D 1 For capecitabine. In some such implementations, D 1 For irinotecan. In some such implementations, D 1 It is trofluuridine. In some such embodiments, D 1 For Doxorubic stars. In some such implementations, D 1 For Dorcetathione. In some such implementations, D 1 It is Infiniti.

[0112] In some implementation schemes, D 1 It is an opioid antagonist. In some embodiments, the opioid antagonist is naloxone, nalmefen, natrozone, methylnaltrozone, nalbuphine, or nalprofen.

[0113] In some implementation schemes, D 1 It is an opioid agonist. In some embodiments, the opioid agonist is morphine, oxycodone, hydromorphone, or methadone.

[0114] In some embodiments, T (a ligand selective for transthyretin-TTR) has the structure of compound (A): Or its pharmaceutically acceptable salt, wherein: X a X b and X c Independently selected from C(R) 8a (R)8b ), O, NR 8b Or S; where R 8a and R 8b Independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted aminoacyl, amino, substituted amino, amide, substituted amide, and cyano; The A ring is a 4- to 12-membered ring. In some embodiments, the 4- to 12-membered ring is an aromatic ring or a heteroaromatic ring. Each Y is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted amino, amino, substituted amino, amide, substituted amide, sulfonamide, sulfonyl fluoride, thioester and cyano; c is an integer in the range of 0 to 5; and Ring B is a heterocycle selected from the following (h1-h30): Where R 11 To R 16 Independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted aminoacyl, amino, substituted amino, amide, substituted amide, and cyano; R 17 Selected from hydroxyl, alkyl, amino, and alkylamino; and R 11 To R 16 At least one of them is related to X c Linking groups.

[0115] In some embodiments, T (a ligand selective for transthyretin-TTR) has the structure of compound (B): (B), Or its pharmaceutically acceptable salt, wherein: n is an integer in the range of 0 to 8; R 18 R 19 and R 20Independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted amino, amino, substituted amino, amide, substituted amide, sulfonamide, sulfonyl fluoride, thioester and cyano; X a For C(R) 21 (R) 22 ), O, NR 22 Or S; where R 21 and R 22 Independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted aminoacyl, amino, substituted amino, amide, substituted amide, and cyano; A is a 5- to 12-membered ring; in some embodiments, the 5- to 12-membered ring is an aromatic ring or a heteroaromatic ring. Each Y is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted amino, amino, substituted amino, amide, substituted amide, sulfonamide, sulfonyl fluoride, thioester, and cyano; and, c is a number from 0 to 5.

[0116] In some embodiments, T (a ligand selective for transthyretin-TTR) has the structure of compound (C): (C), Or its pharmaceutically acceptable salt, wherein: n is an integer in the range of 1 to 4; R 23 It is a short-chain alkyl group having 1 to 4 carbon atoms; R 24 It is hydrogen; R 25 It is a short-chain alkyl group having 1 to 4 carbon atoms; X a For C(R) 21 (R) 22 ), O, NR 22 Or S; where R 21 and R22 Independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted aminoacyl, amino, substituted amino, amide, substituted amide, and cyano; Each Y is independently selected from hydrogen, halogen, acyl, substituted acyl, carboxyl, heterocyclic, alkoxycarbonyl, sulfonamide, sulfonyl fluoride, thioester, and substituted alkoxycarbonyl; and c is 2.

[0117] In some embodiments, the ligand of compound (C) has the following structure, wherein R 23 It is methyl and R 24 Xa is methyl; Xa is O; and Y is fluorine or carboxyl.

[0118] In some embodiments, T (a ligand selective for transthyretin-TTR) has the structure of compound (D): (D), Or its pharmaceutically acceptable salt, wherein: n is between 1 and 8; R 23 R 24 and R 25 Independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted aminoacyl, amino, substituted amino, amide, substituted amide, and cyano; X a For C(R) 21 (R) 22 ), O, NR 22 Or S; where R 21 and R 22 Independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted aminoacyl, amino, substituted amino, amide, substituted amide, and cyano; R a For CHO, COOH, COOCH3, COOR 6 CONR 7R 8 Tetrazolyl, CONHOH, B(OH)2, CONHSO2Ar, CONHCH(R) 9 COOH, CF3, hydrogen, halogen, alkyl, substituted alkyl, acyl, substituted acyl, carboxyl, heterocyclic, sulfonamide, sulfonyl fluoride, thioester, alkoxycarbonyl or substituted alkoxycarbonyl; R b For CHO, COOH, COOCH3, COOR2 6 CONR2 7 R2 8 Tetrazolyl, CONHOH, B(OH)2, CONHSO2Ar, CONHCH(R2) 9 COOH, CF3, hydrogen, halogen, alkyl, substituted alkyl, acyl, substituted acyl, carboxyl, heterocyclic, sulfonamide, sulfonyl fluoride, thioester, alkoxycarbonyl or substituted alkoxycarbonyl; R2 6 It is an alkyl, haloalkyl, cycloalkyl, or heterocyclic group; R 27 and R 28 Each is independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, or heteroaryl; and R 29 It is a side chain of a naturally occurring α-aminocarboxylic acid.

[0119] In some embodiments, the ligand of compound (D) has R b The structure is selected from bromine, chlorine and fluorine.

[0120] In some embodiments, the ligand of compound (D) has the structure of compound (E): (E) Or its pharmaceutically acceptable salt.

[0121] In some embodiments, T (a ligand selective for transthyretin-TTR) has the structure of compound (F): (F) Or its pharmaceutically acceptable salt.

[0122] In some implementations, T (a ligand selective for transthyretin-TTR) has the structure of compound (G): (G) Or its pharmaceutically acceptable salt, wherein R a For OH, CHO, COOH, CONH2, CONH(OH), COOR30 CONHR 30 ; R 30 It is a straight-chain or branched alkyl group with 1-3 carbon atoms.

[0123] In some embodiments, T (a ligand selective for transthyretin-TTR) has the structure of compound (H): (H) Or its pharmaceutically acceptable salt.

[0124] In some embodiments, T (a ligand selective for transthyretin-TTR) has the structure of compound (I): (J) Or its pharmaceutically acceptable salt.

[0125] In some implementations, T (a ligand selective for transthyretin-TTR) has the structure of compound (K): (K) Or its pharmaceutically acceptable salt.

[0126] In some embodiments, T (a ligand selective for transthyretin-TTR) has the structure of compound (L): (L) Or its pharmaceutically acceptable salt.

[0127] In some embodiments, T (a ligand selective for transthyretin-TTR) has the structure of compound (M): (M) Or its pharmaceutically acceptable salt.

[0128] In some embodiments, T (a ligand selective for transthyretin-TTR) has the structure of compound (N): (N) Or its pharmaceutically acceptable salt.

[0129] In some embodiments, T (a ligand selective for transthyretin-TTR) has the structure of compound (O): (O) Or its pharmaceutically acceptable salt.

[0130] In some embodiments, T (a ligand selective for transthyretin-TTR) has the structure of compound (P): (P) Or its pharmaceutically acceptable salt.

[0131] In some implementations, T (a ligand selective for transthyretin-TTR) has the structure of compound (Q): (Q) Or its pharmaceutically acceptable salt.

[0132] In some embodiments, T (a ligand selective for transthyretin-TTR) has the structure of compound (R): (R) Or its pharmaceutically acceptable salt.

[0133] In some implementations, T (a ligand selective for transthyretin-TTR) has the structure of compound (S): (S) Or its pharmaceutically acceptable salt.

[0134] In some implementations, T (a ligand selective for transthyretin TTR) has the structure of compound (T): (T) Or its pharmaceutically acceptable salt.

[0135] In some embodiments, T (a ligand selective for transthyretin-TTR) has the structure of compound (U): (U) Or its pharmaceutically acceptable salt.

[0136] In some embodiments, T (a ligand selective for transthyretin-TTR) has the structure of compound (V): (V) Or its pharmaceutically acceptable salt.

[0137] In some embodiments, T (a ligand selective for transthyretin-TTR) has the structure of compound (W): (W) Or its pharmaceutically acceptable salt.

[0138] In some implementations, T (a ligand selective for transthyretin-TTR) has the structure of compound (Y): (Y) Or its pharmaceutically acceptable salt.

[0139] In some embodiments of compound (Y), the linker (L) is attached to the meta-C16 position of the carboxyl carbon at C14 of the ligand, for example, in the conjugate of formula (Ia): (Ia) In some embodiments of compound (Y), the linker (L) is attached to the C15 position adjacent to the carboxyl carbon of the ligand at C14, for example, in the conjugate of formula (Ib): (Ib) In some implementations, T (a ligand selective for transthyretin-TTR) has one of the following structures: .

[0140] In some implementations, T (a ligand selective for transthyretin-TTR) has one of the following structures: In some embodiments of the conjugates of formula (Ia) or (Ib), L is C 4-20 alkylene, wherein the C 4-20 Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally selected from -O-, -NR. x The - and -S- groups are substituted, where L 1a Halogenated, C 1-6 Alkyl groups, or C groups substituted with halogen groups 1-6 Alkyl group. In some embodiments, L is C. 10-20 alkylene, wherein the C 10-20 Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally replaced by groups selected from -O-, -NH-, -N(CH3)-, and -S-. In some embodiments, L is C 10-15 alkylene, wherein the C 10-15 Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally replaced by groups selected from -O-, -NH-, and -N(CH3)-. In some embodiments, L is C 5-10 alkylene, wherein the C 5-10 Alkylene is optionally surrounded by one or more L 1a The alkylene units are substituted, and one or more alkylene units are optionally replaced by groups selected from -O- and -NH-.

[0141] In some embodiments of formula (Ia) or (Ib), L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion .

[0142] In some implementation schemes, LL B Includes part In some implementations, LL B Includes part In some implementations, LL B Includes part In some implementation schemes, LL B Includes part In some implementations, LL B Includes part In some implementations, LL B Includes part .

[0143] In some embodiments of formula (Ia) or formula (Ib), LL B Includes part or , where R 9 For H or C 1-4 Alkyl, R 10 For H or C 1-4 Alkyl group, and Q is -CH2-, -O-, -S-, -NH- or -N(C 1-4 alkyl)-.

[0144] In one embodiment, this document provides a conjugate of formula (II'): (II'), Or its pharmaceutically acceptable salt, wherein: Each R1 Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n can be 0, 1, 2, 3, or 4; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4 alkyl; L stands for connector; Y 1 For key or C 1-4 Alkylene; Y 2 For NR 6 , O or S; R 6 For H or C 1-4 alkyl; Each R 7 Choose independently from the following groups: hydrogen, halogen atoms (e.g., Cl, Br, or F), NO2, CF3, OMe, O(C 1-4 Alkyl), hydroxyl, -OCF3, -NH2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 cycloalkyl; p is 0, 1, 2, or 3; and D 1 This refers to the pharmaceutical component.

[0145] In one embodiment, this document provides a conjugate of formula (II): (II), Or its pharmaceutically acceptable salt, wherein: Each R 1Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n can be 0, 1, 2, 3, or 4; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4 alkyl; L stands for connector; Y 1 For key or C 1-4 Alkylene; Y 2 For NR 6 , O or S; R 6 For H or C 1-4 alkyl; Each R 7 Choose independently from the following groups: hydrogen, halogen atoms (e.g., Cl, Br, or F), NO2, CF3, OMe, O(C 1-4 Alkyl), hydroxyl, -OCF3, -NH2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 cycloalkyl; p is 0, 1, 2, or 3; and D 1 This refers to the pharmaceutical component.

[0146] In some embodiments, the conjugate of formula (II) has formula (IIa): (IIa) Or its pharmaceutically acceptable salt.

[0147] In some embodiments, the conjugate of formula (II) has formula (IIb): (IIb) Or its pharmaceutically acceptable salt.

[0148] In some embodiments, the formula (II) conjugate has formula (IIc): (IIc) Or its pharmaceutically acceptable salt.

[0149] In some embodiments, the formula (II) conjugate has formula (IId): (IId) Or its pharmaceutically acceptable salt.

[0150] In some embodiments, the formula (II) conjugate has formula (IIe): (IIe) Or its pharmaceutically acceptable salt.

[0151] In some embodiments, the conjugate of formula (II) has formula (IIf): (IIf) Or its pharmaceutically acceptable salt.

[0152] In some embodiments, the formula (II) conjugate has the formula (IIg): (IIg) Or its pharmaceutically acceptable salt.

[0153] In some embodiments, the formula (II) conjugate has formula (IIh): (IIh) Or its pharmaceutically acceptable salt.

[0154] In some embodiments, the conjugate of formula (II) has formula (IIi): (IIi) Or its pharmaceutically acceptable salt.

[0155] In some implementations, the formula (II') conjugate has the formula (IIj): (IIj) Or its pharmaceutically acceptable salt.

[0156] In some implementations, the formula (II') conjugate has the formula (IIk): (IIk) Or its pharmaceutically acceptable salt.

[0157] In some embodiments of the conjugates of formula (II) or (II') or any variation thereof (e.g., formulas (IIa) to (IIk)), R 2 For H or C 1-2 Alkyl group. In some embodiments, R 2 It is H or -CH3. In some implementations, R 2 For H.

[0158] In some embodiments of the conjugates of formula (II) or (II') or any variation thereof (e.g., formulas (IIa) to (IIk)), R 3 For H or C 1-2 Alkyl group. In some embodiments, R 3 It is H or -CH3. In some implementations, R 3 For H.

[0159] In some embodiments of the conjugates of formula (II) or (II') or any variation thereof (e.g., formulas (IIa) to (IIk)), R 4 For H or C 1-2 Alkyl group. In some embodiments, R 4 It is H or -CH3. In some implementations, R 4 It is -CH3.

[0160] In some embodiments of the conjugates of formula (II) or (II') or any variation thereof (e.g., formulas (IIa) to (IIk)), R 5 For H or C 1-2 Alkyl group. In some embodiments, R 5 It is H or -CH3. In some implementations, R 5 It is -CH3.

[0161] In some embodiments of conjugates of formula (II) or (II') or any variation thereof (e.g., formulas (IIa) to (IIh), (IIj) or (IIk)), n is 2 or 3. In some embodiments, n is 3.

[0162] In some embodiments of conjugates of formula (II) or (II') or any variation thereof (e.g., formulas (IIa) to (IIh) or (IIj)), m is 0, 1, or 2. In some embodiments, m is 0 or 1. In some embodiments, m is 0. In some embodiments, m is 1.

[0163] In some embodiments of conjugates of formula (II) or (II') or any variation thereof (e.g., formulas (IIa) to (IIh) or (IIj)), p is 0, 1, or 2. In some embodiments, p is 0 or 1. In some embodiments, p is 0. In some embodiments, p is 1.

[0164] In some embodiments of the conjugates of formula (II) or (II') or any variation thereof (e.g., formulas (IIa) to (IIk)), L is C 4-20 alkylene, wherein the C 4-20 Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally selected from -O-, -NR. x The - and -S- groups are substituted, where L 1a Halogenated, C 1-6 Alkyl groups, or C groups substituted with halogen groups 1-6 Alkyl group. In some embodiments, L is C. 10-20 alkylene, wherein the C 10-20 Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally replaced by groups selected from -O-, -NH-, -N(CH3)-, and -S-. In some embodiments, L is C 10-15 alkylene, wherein the C 10-15 Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally replaced by groups selected from -O-, -NH-, and -N(CH3)-. In some embodiments, L is C 5-10 alkylene, wherein the C 5-10 Alkylene is optionally surrounded by one or more L 1a The alkylene units are substituted, and one or more alkylene units are optionally replaced by groups selected from -O- and -NH-.

[0165] In some embodiments of the conjugate of formula (II) or (II') or any variation thereof (e.g., formulas (IIa) to (IIi)), L contains a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion .

[0166] In some embodiments of the conjugates of formula (II) or (II') or any variation thereof (e.g., formulas (IIa) to (IIk)), D 1 For proteins. In some implementations, D 1 The cytokine is used. In some embodiments, the cytokine is interleukin-2 (IL-2). In some embodiments, the cytokine is mutant IL-2, such as the cytokine described in US 9,206,943. In some embodiments, the cytokine is interleukin-15 (IL-15). In some embodiments, the cytokine is mutant IL-15. In one such embodiment, IL-15 is IL-15[N27D]. In some embodiments, the cytokine is interleukin-7 (IL-7). In some embodiments, the cytokine is interleukin-9 (IL-9). In some embodiments, the cytokine is interleukin-10 (IL-10). In some embodiments, the cytokine is interleukin-21 (IL-21). In some embodiments, the cytokine is interleukin-10 (IL-10).

[0167] In some embodiments of the conjugates of formula (II) or (II') or any variation thereof (e.g., formulas (IIa) to (IIk)), D 1 It is an anticancer agent. In some such implementations, D 1 For SN-38. In some such implementations, D 1 It is monomethylolpropionate E. In some such embodiments, D 1 For capecitabine. In some such implementations, D 1 For irinotecan. In some such implementations, D 1 It is trofluuridine. In some such embodiments, D 1 For Doxorubic stars. In some such implementations, D 1 For Dorcetathione. In some such implementations, D 1 It is Infiniti.

[0168] In some embodiments of the conjugates of formula (II) or (II') or any variation thereof (e.g., formulas (IIa) to (IIk)), D 1 It is a peptide.

[0169] In some embodiments, the peptide is RGK-MCA. In some embodiments, the peptide is neurotensin. In some embodiments, the peptide is gonadotropin-releasing hormone (GnRH). In some embodiments, the peptide is GIP (1-42) human peptide. In some embodiments, the peptide is D6-GnRH. In some embodiments, the peptide is insulin. In some embodiments, the peptide is teriparatide. In some embodiments, the peptide is salmon calcitonin. In some embodiments, the peptide is iparene. In some embodiments, the peptide is vasopressin. In some embodiments, the peptide is glucagon. In some embodiments, the peptide is alexiside. In some embodiments, the peptide is goserelin. In some embodiments, the peptide is leuprolide. In some embodiments, the peptide is teriparatide. In some embodiments, the peptide is lixinaside. In some embodiments, the peptide is desmopressin. In some embodiments, the peptide is oxytocin. In some embodiments, the peptide is corticotropin. In some embodiments, the peptide is sermorelin. In some embodiments, the peptide is bumenotide. In some embodiments, the peptide is abalotropic peptide. In some embodiments, the peptide is dilorelin.

[0170] In some embodiments, the peptide drug is a glucagon-like peptide-1 (GLP-1) receptor agonist used to treat obesity and / or diabetes. In some such embodiments, the peptide drug is semaglutide, liraglutide, dulaglutide, abiglutide, or exenatide. Because the duration of action of the peptide drug is significantly prolonged when incorporated into the conjugates of this disclosure, GLP-1 agonist conjugates can reduce the frequency of administration, such as once weekly, once every two weeks, once every three weeks, or once monthly.

[0171] In some embodiments, the peptide drug is a glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic peptide (GIP) receptor agonist used to treat obesity and / or diabetes. In some such embodiments, the peptide drug is telposide. Because the duration of action of the peptide drug is significantly prolonged when incorporated into the conjugates of this disclosure, the GLP-1 / GIP agonist conjugates can reduce the frequency of administration, such as once weekly, once every two weeks, once every three weeks, or once monthly.

[0172] In some embodiments, the peptide is an endogenous opioid peptide. In some such embodiments, the peptide is endorphin-1 or endorphin-2. In other embodiments, the peptide is corticomerol.

[0173] In some embodiments of the conjugates of formula (II) or (II') or any variation thereof (e.g., formulas (IIa) to (IIk)), D 1It is an opioid antagonist. In some embodiments, the opioid antagonist is naloxone, nalmefen, natrozone, methylnaltrozone, nalbuphine, or nalprofen.

[0174] In some embodiments of the conjugates of formula (II) or (II') or any variation thereof (e.g., formulas (IIa) to (IIk)), D 1 It is an opioid agonist. In some embodiments, the opioid agonist is morphine, oxycodone, hydromorphone, or methadone.

[0175] In one embodiment, this document provides a conjugate of formula (III'): (III') Or its pharmaceutically acceptable salt, wherein: Each R 1 Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n can be 0, 1, 2, 3, or 4; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4 alkyl; L A C 3-20 Alkylene, wherein The C 3-20 Alkylene is optionally surrounded by one or more L 1a Replace, and One or more alkylene units are optionally selected from -O-, -NR y Substitution of - and -S- groups; Each L 1a Halogenated, C 1-6 Alkyl groups, or C groups substituted with halogen groups 1-6 alkyl; Each R y For H or C 1-4 alkyl; L BThe click product is formed by the click reaction between the first click handle and the second click handle; Y 1 For key or C 1-4 Alkylene; Y 2 For NR 6 , O or S; R 6 For H or C 1-4 alkyl; Each R 7 Choose independently from the following groups: hydrogen, halogen atoms (e.g., Cl, Br, or F), NO2, CF3, OMe, O(C 1-4 Alkyl), hydroxyl, -OCF3, -NH2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 cycloalkyl; p is 0, 1, 2, or 3; and D 1 This refers to the pharmaceutical component.

[0176] In one embodiment, this document provides a conjugate of formula (III): (III) Or its pharmaceutically acceptable salt, wherein: Each R 1 Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n can be 0, 1, 2, 3, or 4; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4alkyl; L A C 3-20 Alkylene, wherein The C 3-20 Alkylene is optionally surrounded by one or more L 1a Replace, and One or more alkylene units are optionally selected from -O-, -NR y Substitution of - and -S- groups; Each L 1a Halogenated, C 1-6 Alkyl groups, or C groups substituted with halogen groups 1-6 alkyl; Each R y For H or C 1-4 alkyl; L B The click product is formed by the click reaction between the first click handle and the second click handle; Y 1 For key or C 1-4 Alkylene; Y 2 For NR 6 , O or S; R 6 For H or C 1-4 alkyl; Each R 7 Choose independently from the following groups: hydrogen, halogen atoms (e.g., Cl, Br, or F), NO2, CF3, OMe, O(C 1-4 Alkyl), hydroxyl, -OCF3, -NH2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 cycloalkyl; p is 0, 1, 2, or 3; and D 1 This refers to the pharmaceutical component.

[0177] In some embodiments, the formula (III) conjugate has formula (IIIa): (IIIa) Or its pharmaceutically acceptable salt.

[0178] In some embodiments, the formula (III) conjugate has formula (IIIb): (IIIb) Or its pharmaceutically acceptable salt.

[0179] In some embodiments, the formula (III) conjugate has formula (IIIc): (IIIc) Or its pharmaceutically acceptable salt.

[0180] In some embodiments, the formula (III) conjugate has formula (IIId): (IIId) Or its pharmaceutically acceptable salt.

[0181] In some embodiments, the formula (III) conjugate has formula (IIIe): (IIIe) Or its pharmaceutically acceptable salt.

[0182] In some embodiments, the formula (III) conjugate has formula (IIIf): (IIIf) Or its pharmaceutically acceptable salt.

[0183] In some embodiments, the formula (III) conjugate has the formula (IIIg): (IIIg) Or its pharmaceutically acceptable salt.

[0184] In some embodiments, the formula (III) conjugate has formula (IIIh): (IIIh) Or its pharmaceutically acceptable salt.

[0185] In some embodiments, the formula (III) conjugate has formula (IIIi): (IIIi) Or its pharmaceutically acceptable salt.

[0186] In some implementations, the (III') conjugate has the formula (IIIj): (IIIj) Or its pharmaceutically acceptable salt.

[0187] In some implementations, the (III') conjugate has the formula (IIIk): (IIIk) Or its pharmaceutically acceptable salt.

[0188] In some embodiments of the conjugates of formula (III) or (III') or any variation thereof (e.g., formulas (IIIa) to (IIIk)), R 2 For H or C 1-2 Alkyl group. In some embodiments, R 2 It is H or -CH3. In some implementations, R 2 For H.

[0189] In some embodiments of the conjugates of formula (III) or (III') or any variation thereof (e.g., formulas (IIIa) to (IIIk)), R 3 For H or C 1-2 Alkyl group. In some embodiments, R 3 It is H or -CH3. In some implementations, R 3 For H.

[0190] In some embodiments of the conjugates of formula (III) or (III') or any variation thereof (e.g., formulas (IIIa) to (IIIk)), R 4 For H or C 1-2 Alkyl group. In some embodiments, R 4 It is H or -CH3. In some implementations, R 4 It is -CH3.

[0191] In some embodiments of the conjugates of formula (III) or (III') or any variation thereof (e.g., formulas (IIIa) to (IIIk)), R 5 For H or C 1-2 Alkyl group. In some embodiments, R 5 It is H or -CH3. In some implementations, R 5 It is -CH3.

[0192] In some embodiments of conjugates of formula (III) or (III') or any variation thereof (e.g., formulas (IIIa) to (IIIk)), n is 2 or 3. In some embodiments, n is 3.

[0193] In some embodiments of conjugates of formula (III) or (III') or any variation thereof (e.g., formulas (IIIa) to (IIIj)), m is 0, 1, or 2. In some embodiments, m is 0 or 1. In some embodiments, m is 0. In some embodiments, m is 1.

[0194] In some embodiments of conjugates of formula (III) or (III') or any variation thereof (e.g., formulas (IIIa), (IIIb), or (IIId) through (IIIj)), p is 0, 1, or 2. In some embodiments, p is 0 or 1. In some embodiments, p is 0. In some embodiments, p is 1.

[0195] In some embodiments of the conjugates of formula (III) or (III') or any variation thereof (e.g., formulas (IIIa) to (IIIk)), L A C 4-20 alkylene, wherein the C 4-20 Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally selected from -O-, -NR. x The - and -S- groups are substituted, where L 1a Halogenated, C 1-6 Alkyl groups, or C groups substituted with halogen groups 1-6 Alkyl group, wherein R x H or C optionally substituted with a halogen group 1-6 Alkyl group. In some embodiments, L A C 10-20 alkylene, wherein the C 10-20 Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally replaced by groups selected from -O-, -NH-, -N(CH3)-, and -S-. In some embodiments, L A C 10-15 alkylene, wherein the C 10-15 Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally replaced by groups selected from -O-, -NH-, and -N(CH3)-. In some embodiments, L A C 5-10 alkylene, wherein the C 5-10 Alkylene is optionally surrounded by one or more L 1a The alkylene units are substituted, and one or more alkylene units are optionally replaced by groups selected from -O- and -NH-.

[0196] In some embodiments of the conjugates of formula (III) or (III') or any variation thereof (e.g., formulas (IIIa) to (IIIk)), L A Includes part In some implementations, L A Includes part In some implementations, L AIncludes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion .

[0197] In some embodiments of the conjugates of formula (III) or (III') or any variation thereof (e.g., formulas (IIIa) to (IIIk)), the click product L B The triazine moiety. In some embodiments, the conjugate of any of formula (III) or (III'), or formulas (IIIa) to (IIIi), comprises one of the following moieties: , where X 2 -CH2-, -NR x -, -S-, or -O-, where R x As defined elsewhere herein. In some embodiments, the conjugate comprises a portion In some implementations, the conjugate comprises a portion .

[0198] In some embodiments of the conjugates of formula (III) or (III') or any variation thereof (e.g., formulas (IIIa) to (IIIk)), L A -L B Includes part In some implementations, L A -L B Includes part In some implementations, L A -L B Includes part In some implementations, L A -L B Includes part In some implementations, L A -L BIncludes part In some implementations, L A -L B Includes part .

[0199] In some embodiments of the conjugates of formula (III) or (III') or any variation thereof (e.g., formulas (IIIa) to (IIIk)), L A -L B Includes part or , where R 9 For H or C 1-4 Alkyl, R 10 For H or C 1-4 Alkyl group, and Q is -CH2-, -O-, -S-, -NH- or -N(C 1-4 alkyl)-.

[0200] In some embodiments of the conjugates of formula (III) or (III') or any variation thereof (e.g., formulas (IIIa) to (IIIk)), D 1 For proteins. In some implementations, D 1 The cytokine is used. In some embodiments, the cytokine is interleukin-2 (IL-2). In some embodiments, the cytokine is mutant IL-2, such as the cytokine described in US 9,206,943. In some embodiments, the cytokine is interleukin-15 (IL-15). In some embodiments, the cytokine is mutant IL-15. In one such embodiment, IL-15 is IL-15[N27D]. In some embodiments, the cytokine is interleukin-7 (IL-7). In some embodiments, the cytokine is interleukin-9 (IL-9). In some embodiments, the cytokine is interleukin-10 (IL-10). In some embodiments, the cytokine is interleukin-21 (IL-21). In some embodiments, the cytokine is interleukin-10 (IL-10).

[0201] In some embodiments of the conjugates of formula (III) or (III') or any variation thereof (e.g., formulas (IIIa) to (IIIk)), D 1The peptide is a peptide. In some embodiments, the peptide is RGK-MCA. In some embodiments, the peptide is neurotensin. In some embodiments, the peptide is gonadotropin-releasing hormone (GnRH). In some embodiments, the peptide is GIP (1-42) human peptide. In some embodiments, the peptide is D6-GnRH. In some embodiments, the peptide is insulin. In some embodiments, the peptide is teriparatide. In some embodiments, the peptide is salmon calcitonin. In some embodiments, the peptide is iparene. In some embodiments, the peptide is vasopressin. In some embodiments, the peptide is glucagon. In some embodiments, the peptide is alexiside. In some embodiments, the peptide is goserelin. In some embodiments, the peptide is leuprolide. In some embodiments, the peptide is teriparatide. In some embodiments, the peptide is lixinaside. In some embodiments, the peptide is desmopressin. In some embodiments, the peptide is oxytocin. In some embodiments, the peptide is corticotropin. In some embodiments, the peptide is sermorelin. In some embodiments, the peptide is bumenotide. In some embodiments, the peptide is abalotropic peptide. In some embodiments, the peptide is dilorelin.

[0202] In some embodiments, the peptide drug is a glucagon-like peptide-1 (GLP-1) receptor agonist used to treat obesity and / or diabetes. In some such embodiments, the peptide drug is semaglutide, liraglutide, dulaglutide, abiglutide, or exenatide. Because the duration of action of the peptide drug is significantly prolonged when incorporated into the conjugates of this disclosure, GLP-1 agonist conjugates can reduce the frequency of administration, such as once weekly, once every two weeks, once every three weeks, or once monthly.

[0203] In some embodiments, the peptide drug is a glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic peptide (GIP) receptor agonist used to treat obesity and / or diabetes. In some such embodiments, the peptide drug is telposide. Because the duration of action of the peptide drug is significantly prolonged when incorporated into the conjugates of this disclosure, the GLP-1 / GIP agonist conjugates can reduce the frequency of administration, such as once weekly, once every two weeks, once every three weeks, or once monthly.

[0204] In some embodiments, the peptide is an endogenous opioid peptide. In some such embodiments, the peptide is endorphin-1 or endorphin-2. In other embodiments, the peptide is corticomerol.

[0205] In some embodiments of the conjugates of formula (III) or (III') or any variation thereof (e.g., formulas (IIIa) to (IIIk)), D 1 It is an anticancer agent. In some such implementations, D 1For SN-38. In some such implementations, D 1 It is monomethylolpropionate E. In some such embodiments, D 1 For capecitabine. In some such implementations, D 1 For irinotecan. In some such implementations, D 1 It is trofluuridine. In some such embodiments, D 1 For Doxorubic stars. In some such implementations, D 1 For Dorcetathione. In some such implementations, D 1 It is Infiniti.

[0206] In some embodiments of the conjugates of formula (III) or (III') or any variation thereof (e.g., formulas (IIIa) to (IIIk)), D 1 It is an opioid antagonist. In some embodiments, the opioid antagonist is naloxone, nalmefen, natrozone, methylnaltrozone, nalbuphine, or nalprofen.

[0207] In some embodiments of the conjugates of formula (III) or (III') or any variation thereof (e.g., formulas (IIIa) to (IIIk)), D 1 It is an opioid agonist. In some embodiments, the opioid agonist is morphine, oxycodone, hydromorphone, or methadone.

[0208] In one embodiment, this document provides a (IV') conjugate: (IV') Or its pharmaceutically acceptable salt, wherein: Each R 1 Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n can be 0, 1, 2, 3, or 4; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4 alkyl; L A C 3-20 Alkylene, wherein The C 3-20 Alkylene is optionally surrounded by one or more L 1a Replace, and One or more alkylene units are optionally selected from -O-, -NR y Substitution of - and -S- groups; Each L 1a Halogenated, C 1-6 Alkyl groups, or C groups substituted with halogen groups 1-6 alkyl; Each R y For H or C 1-4 alkyl; L B The click product is formed by the click reaction between the first click handle and the second click handle; Y 1 For key or C 1-4 Alkylene; Y 2 For NR 6 , O or S; R 6 For H or C 1-4 alkyl; Each R 7 Choose independently from the following groups: hydrogen, halogen atoms (e.g., Cl, Br, or F), NO2, CF3, OMe, O(C 1-4 Alkyl), hydroxyl, -OCF3, -NH2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 Cycloalkyl; p is 0, 1, 2 or 3; and D 1 This refers to the pharmaceutical component.

[0209] In one embodiment, this document provides a formula (IV) conjugate: (IV) Or its pharmaceutically acceptable salt, wherein: Each R 1 Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n can be 0, 1, 2, 3, or 4; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4 alkyl; L A C 3-20 Alkylene, wherein The C 3-20 Alkylene is optionally surrounded by one or more L 1a Replace, and One or more alkylene units are optionally selected from -O-, -NR y Substitution of - and -S- groups; Each L 1a Halogenated, C 1-6 Alkyl groups, or C groups substituted with halogen groups 1-6 alkyl; Each R y For H or C 1-4 alkyl; L B The click product is formed by the click reaction between the first click handle and the second click handle; Y 1 For key or C 1-4 Alkylene; Y 2 For NR 6 , O or S; R 6 For H or C 1-4 alkyl; Each R 7 Choose independently from the following groups: hydrogen, halogen atoms (e.g., Cl, Br, or F), NO2, CF3, OMe, O(C 1-4 Alkyl), hydroxyl, -OCF3, -NH2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 Cycloalkyl; p is 0, 1, 2 or 3; and D 1 This refers to the pharmaceutical component.

[0210] In some implementations, the (IV) conjugate has the formula (IVa): (IVa) Or its pharmaceutically acceptable salt.

[0211] In some implementations, the (IV) conjugate has the formula (IVb): (IVb) Or its pharmaceutically acceptable salt.

[0212] In some implementations, the (IV) conjugate has the formula (IVc): (IVc) Or its pharmaceutically acceptable salt.

[0213] In some implementations, the (IV) conjugate has the formula (IVd): Or its pharmaceutically acceptable salt.

[0214] In some implementations, the (IV) conjugate has the formula (IVe): (IVe) Or its pharmaceutically acceptable salt.

[0215] In some implementations, the (IV) conjugate has the formula (IVf): (IVf) Or its pharmaceutically acceptable salt.

[0216] In some implementations, the (IV) conjugate has the formula (IVg): (IVg) Or its pharmaceutically acceptable salt.

[0217] In some implementations, the (IV) conjugate has the formula (IVh): (IVh) Or its pharmaceutically acceptable salt.

[0218] In some implementations, the (IV) conjugate has the formula (IVi): (IVi) Or its pharmaceutically acceptable salt.

[0219] In some implementations, the (IV') conjugate has the formula (IVj): (IVj) Or its pharmaceutically acceptable salt.

[0220] In some implementations, the (IV') conjugate has the formula (IVk): (IVk) Or its pharmaceutically acceptable salt.

[0221] In some embodiments of the conjugates of formula (IV) or (IV') or any variation thereof (e.g., formulas (IVa) to (IVk)), R 2 For H or C 1-2 Alkyl group. In some embodiments, R 2 It is H or -CH3. In some implementations, R 2 For H.

[0222] In some embodiments of the conjugates of formula (IV) or (IV') or any variation thereof (e.g., formulas (IVa) to (IVk)), R 3 For H or C 1-2 Alkyl group. In some embodiments, R 3 It is H or -CH3. In some implementations, R 3 For H.

[0223] In some embodiments of the conjugates of formula (IV) or (IV') or any variation thereof (e.g., formulas (IVa) to (IVk)), R 4 For H or C 1-2 Alkyl group. In some embodiments, R 4 It is H or -CH3. In some implementations, R 4 It is -CH3.

[0224] In some embodiments of the conjugates of formula (IV) or (IV') or any variation thereof (e.g., formulas (IVa) to (IVk)), R 5 For H or C 1-2 Alkyl group. In some embodiments, R 5 It is H or -CH3. In some implementations, R 5 It is -CH3.

[0225] In some embodiments of conjugates of formula (IV) or (IV') or any variation thereof (e.g., formulas (IVa) to (IVk)), n is 2 or 3. In some embodiments, n is 3.

[0226] In some embodiments of conjugates of formula (IV) or (IV') or any variation thereof (e.g., formulas (IVa) to (IVj)), m is 0, 1, or 2. In some embodiments, m is 0 or 1. In some embodiments, m is 0. In some embodiments, m is 1.

[0227] In some embodiments of conjugates of formula (IV) or (IV') or any variation thereof (e.g., formulas (IVa) to (IVj)), p is 0, 1, or 2. In some embodiments, p is 0 or 1. In some embodiments, p is 0. In some embodiments, p is 1.

[0228] In some embodiments of the conjugate of formula (IV) or (IV') or any variation thereof (e.g., formulas (IVa) to (IVk)), L A C 4-20 alkylene, wherein the C 4-20 Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally selected from -O-, -NR. x The - and -S- groups are substituted, where L 1a Halogenated, C 1-6 Alkyl groups, or C groups substituted with halogen groups 1-6 Alkyl group, wherein R x H or C optionally substituted with a halogen group 1-6 Alkyl group. In some embodiments, L A C 10-20 alkylene, wherein the C 10-20 Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally replaced by groups selected from -O-, -NH-, -N(CH3)-, and -S-. In some embodiments, L A C 10-15 alkylene, wherein the C 10-15 Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally replaced by groups selected from -O-, -NH-, and -N(CH3)-. In some embodiments, L A C 5-10 alkylene, wherein the C 5-10 Alkylene is optionally surrounded by one or more L 1aThe alkylene units are substituted, and one or more alkylene units are optionally replaced by groups selected from -O- and -NH-.

[0229] In some embodiments of the conjugate of formula (IV) or (IV') or any variation thereof (e.g., formulas (IVa) to (IVk)), L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part .

[0230] In some embodiments of the conjugates of formula (IV) or (IV') or any variation thereof (e.g., formulas (IVa) to (IVk)), D 1 For proteins. In some implementations, D 1The cytokine is used. In some embodiments, the cytokine is interleukin-2 (IL-2). In some embodiments, the cytokine is mutant IL-2, such as the cytokine described in US 9,206,943. In some embodiments, the cytokine is interleukin-15 (IL-15). In some embodiments, the cytokine is mutant IL-15. In one such embodiment, IL-15 is IL-15[N27D]. In some embodiments, the cytokine is interleukin-7 (IL-7). In some embodiments, the cytokine is interleukin-9 (IL-9). In some embodiments, the cytokine is interleukin-10 (IL-10). In some embodiments, the cytokine is interleukin-21 (IL-21). In some embodiments, the cytokine is interleukin-10 (IL-10).

[0231] In some embodiments of the conjugates of formula (IV) or (IV') or any variation thereof (e.g., formulas (IVa) to (IVk)), D 1 It is an anticancer agent. In some such implementations, D 1 For SN-38. In some such implementations, D 1 It is monomethylolpropionate E. In some such embodiments, D 1 For capecitabine. In some such implementations, D 1 For irinotecan. In some such implementations, D 1 It is trofluuridine. In some such embodiments, D 1 For Doxorubic stars. In some such implementations, D 1 For Dorcetathione. In some such implementations, D 1 It is Infiniti.

[0232] In some embodiments of the conjugates of formula (IV) or (IV') or any variation thereof (e.g., formulas (IVa) to (IVk)), D 1 It is a peptide.

[0233] In some embodiments, the peptide is RGK-MCA. In some embodiments, the peptide is neurotensin. In some embodiments, the peptide is gonadotropin-releasing hormone (GnRH). In some embodiments, the peptide is GIP (1-42) human peptide. In some embodiments, the peptide is D6-GnRH. In some embodiments, the peptide is insulin. In some embodiments, the peptide is teriparatide. In some embodiments, the peptide is salmon calcitonin. In some embodiments, the peptide is iparene. In some embodiments, the peptide is vasopressin. In some embodiments, the peptide is glucagon. In some embodiments, the peptide is alexiside. In some embodiments, the peptide is goserelin. In some embodiments, the peptide is leuprolide. In some embodiments, the peptide is teriparatide. In some embodiments, the peptide is lixinaside. In some embodiments, the peptide is desmopressin. In some embodiments, the peptide is oxytocin. In some embodiments, the peptide is corticotropin. In some embodiments, the peptide is sermorelin. In some embodiments, the peptide is bumenotide. In some embodiments, the peptide is abalotropic peptide. In some embodiments, the peptide is dilorelin.

[0234] In some embodiments, the peptide drug is a glucagon-like peptide-1 (GLP-1) receptor agonist used to treat obesity and / or diabetes. In some such embodiments, the peptide drug is semaglutide, liraglutide, dulaglutide, abiglutide, or exenatide. Because the duration of action of the peptide drug is significantly prolonged when incorporated into the conjugates of this disclosure, GLP-1 agonist conjugates can reduce the frequency of administration, such as once weekly, once every two weeks, once every three weeks, or once monthly.

[0235] In some embodiments, the peptide drug is a glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic peptide (GIP) receptor agonist used to treat obesity and / or diabetes. In some such embodiments, the peptide drug is telposide. Because the duration of action of the peptide drug is significantly prolonged when incorporated into the conjugates of this disclosure, the GLP-1 / GIP agonist conjugates can reduce the frequency of administration, such as once weekly, once every two weeks, once every three weeks, or once monthly.

[0236] In some embodiments, the peptide is an endogenous opioid peptide. In some such embodiments, the peptide is endorphin-1 or endorphin-2. In other embodiments, the peptide is corticomerol.

[0237] In some embodiments of the conjugates of formula (IV) or (IV') or any variation thereof (e.g., formulas (IVa) to (IVk)), D 1 It is an opioid antagonist. In some embodiments, the opioid antagonist is naloxone, nalmefen, natrozone, methylnaltrozone, nalbuphine, or nalprofen.

[0238] In some embodiments of the conjugates of formula (IV) or (IV') or any variation thereof (e.g., formulas (IVa) to (IVk)), D 1 It is an opioid agonist. In some embodiments, the opioid agonist is morphine, oxycodone, hydromorphone, or methadone.

[0239] In some implementations, this document provides a conjugate of formula (V'): (V'), Or its pharmaceutically acceptable salt, wherein: Each R 1 Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n is 0, 1, 2, or 3; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4 alkyl; L A C 3-20 Alkylene, wherein The C 3-20 Alkylene is optionally surrounded by one or more L 1a Replace, and One or more alkylene units are optionally selected from -O-, -NR y Substitution of - and -S- groups; Each L 1a Halogenated, C 1-6 Alkyl groups, or C groups substituted with halogen groups 1-6 alkyl; Each R y For H or C 1-4 alkyl; Y 1 For key or C 1-4 Alkylene; Y 2 For NR 6, O or S; R 6 For H or C 1-4 alkyl; Each R 7 Independently selected from the following groups: hydrogen, halogen atoms (e.g., Cl, Br, or F), hydroxyl group, -OCF3, -NH2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 Cycloalkyl; p is 0, 1, 2 or 3; and D 1 This refers to the pharmaceutical component.

[0240] In some implementations, this document provides a conjugate of formula (V): (V), Or its pharmaceutically acceptable salt, wherein: Each R 1 Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n is 0, 1, 2, or 3; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4 alkyl; L A C 3-20 Alkylene, wherein The C 3-20 Alkylene is optionally surrounded by one or more L 1a Replace, and One or more alkylene units are optionally selected from -O-, -NR y Substitution of - and -S- groups; Each L 1a Halogenated, C 1-6 Alkyl groups, or C groups substituted with halogen groups 1-6 alkyl; Each R y For H or C 1-4 alkyl; Y 1 For key or C 1-4 Alkylene; Y 2 For NR 6 , O or S; R 6 For H or C 1-4 alkyl; Each R 7 Independently selected from the following groups: hydrogen, halogen atoms (e.g., Cl, Br, or F), hydroxyl group, -OCF3, -NH2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 Cycloalkyl; p is 0, 1, 2 or 3; and D 1 This refers to the pharmaceutical component.

[0241] In some implementations, the formula (V) conjugate has the formula (Va): (Va) Or its pharmaceutically acceptable salt.

[0242] In some implementations, the (V) conjugate has the formula (Vb): (Vb) Or its pharmaceutically acceptable salt.

[0243] In some implementations, the formula (V) conjugate has the formula (Vc): (Vc) Or its pharmaceutically acceptable salt.

[0244] In some implementations, the formula (V) conjugate has the formula (Vd): (Vd) Or its pharmaceutically acceptable salt.

[0245] In some implementations, the (V) conjugate has the formula (Ve): (Ve) Or its pharmaceutically acceptable salt.

[0246] In some implementations, the (V) conjugate has the formula (Vf): (Vf) Or its pharmaceutically acceptable salt.

[0247] In some implementations, the (V) conjugate has the formula (Vg): (Vg) Or its pharmaceutically acceptable salt.

[0248] In some implementations, the (V') conjugate has the formula (Vh): (Vh) Or its pharmaceutically acceptable salt.

[0249] In some implementations, the formula (V') conjugate has the formula (Vi): (Vi) Or its pharmaceutically acceptable salt.

[0250] In some embodiments of the conjugates of formula (V) or (V') or any variation thereof (e.g., formulas (Va) to (Vi)), R 2 For H or C 1-2 Alkyl group. In some embodiments, R 2 It is H or -CH3. In some implementations, R 2 For H.

[0251] In some embodiments of the conjugates of formula (V) or (V') or any variation thereof (e.g., formulas (Va) to (Vi)), R 3 For H or C 1-2 Alkyl group. In some embodiments, R 3 It is H or -CH3. In some implementations, R 3 For H.

[0252] In some embodiments of the conjugates of formula (V) or (V') or any variation thereof (e.g., formulas (Va) to (Vi)), R 4 For H or C 1-2 Alkyl group. In some embodiments, R 4 It is H or -CH3. In some implementations, R 4 It is -CH3.

[0253] In some embodiments of the conjugates of formula (V) or (V') or any variation thereof (e.g., formulas (Va) to (Vi)), R 5 For H or C 1-2 Alkyl group. In some embodiments, R 5 It is H or -CH3. In some implementations, R 5 It is -CH3.

[0254] In some embodiments of conjugates of formula (V) or (V') or any variation thereof (e.g., formulas (Va) to (Vi)), n is 2 or 3. In some embodiments, n is 3.

[0255] In some embodiments of conjugates of formula (V) or (V') or any variation thereof (e.g., formulas (Va) to (Vi)), m is 0, 1, or 2. In some embodiments, m is 0 or 1. In some embodiments, m is 0. In some embodiments, m is 1.

[0256] In some embodiments of conjugates of formula (V) or (V') or any variation thereof (e.g., formulas (Va) to (Vh)), p is 0, 1, or 2. In some embodiments, p is 0 or 1. In some embodiments, p is 0. In some embodiments, p is 1.

[0257] In some embodiments of the conjugates of formula (V) or (V') or any variation thereof (e.g., formulas (Va) to (Vi)), L A C 4-20 alkylene, wherein the C 4-20 Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally selected from -O-, -NR. x The - and -S- groups are substituted, where L 1a Halogenated, C 1-6 Alkyl groups, or C groups substituted with halogen groups 1-6 Alkyl group, wherein R x H or C optionally substituted with a halogen group 1-6 Alkyl group. In some embodiments, L A C 10-20 alkylene, wherein the C 10-20 Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally replaced by groups selected from -O-, -NH-, -N(CH3)-, and -S-. In some embodiments, L A C 10-15 alkylene, wherein the C 10-15 Alkylene is optionally surrounded by one or more L 1aSubstitution, and one or more alkylene units are optionally replaced by groups selected from -O-, -NH-, and -N(CH3)-. In some embodiments, L A C 5-10 alkylene, wherein the C 5-10 Alkylene is optionally surrounded by one or more L 1a The alkylene units are substituted, and one or more alkylene units are optionally replaced by groups selected from -O- and -NH-.

[0258] In some embodiments of the conjugates of formula (V) or (V') or any variation thereof (e.g., formulas (Va) to (Vi)), L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part .

[0259] In some embodiments of the conjugates of formula (V) or (V') or any variation thereof (e.g., formulas (Va) to (Vi)), D 1 For proteins. In some implementations, D 1The cytokine is used. In some embodiments, the cytokine is interleukin-2 (IL-2). In some embodiments, the cytokine is mutant IL-2, such as the cytokine described in US 9,206,943. In some embodiments, the cytokine is interleukin-15 (IL-15). In some embodiments, the cytokine is mutant IL-15. In one such embodiment, IL-15 is IL-15[N27D]. In some embodiments, the cytokine is interleukin-7 (IL-7). In some embodiments, the cytokine is interleukin-9 (IL-9). In some embodiments, the cytokine is interleukin-10 (IL-10). In some embodiments, the cytokine is interleukin-21 (IL-21). In some embodiments, the cytokine is interleukin-10 (IL-10).

[0260] In some embodiments of the conjugates of formula (V) or (V') or any variation thereof (e.g., formulas (Va) to (Vi)), D 1 It is an anticancer agent. In some such implementations, D 1 For SN-38. In some such implementations, D 1 It is monomethylolpropionate E. In some such embodiments, D 1 For capecitabine. In some such implementations, D 1 For irinotecan. In some such implementations, D 1 It is trofluuridine. In some such embodiments, D 1 For Doxorubic stars. In some such implementations, D 1 For Dorcetathione. In some such implementations, D 1 It is Infiniti.

[0261] In some embodiments of the conjugates of formula (V) or (V') or any variation thereof (e.g., formulas (Va) to (Vi)), D 1 It is a peptide.

[0262] In some embodiments, the peptide is RGK-MCA. In some embodiments, the peptide is neurotensin. In some embodiments, the peptide is gonadotropin-releasing hormone (GnRH). In some embodiments, the peptide is GIP (1-42) human peptide. In some embodiments, the peptide is D6-GnRH. In some embodiments, the peptide is insulin. In some embodiments, the peptide is teriparatide. In some embodiments, the peptide is salmon calcitonin. In some embodiments, the peptide is iparene. In some embodiments, the peptide is vasopressin. In some embodiments, the peptide is glucagon. In some embodiments, the peptide is alexiside. In some embodiments, the peptide is goserelin. In some embodiments, the peptide is leuprolide. In some embodiments, the peptide is teriparatide. In some embodiments, the peptide is lixinaside. In some embodiments, the peptide is desmopressin. In some embodiments, the peptide is oxytocin. In some embodiments, the peptide is corticotropin. In some embodiments, the peptide is sermorelin. In some embodiments, the peptide is bumenotide. In some embodiments, the peptide is abalotropic peptide. In some embodiments, the peptide is dilorelin.

[0263] In some embodiments, the peptide drug is a glucagon-like peptide-1 (GLP-1) receptor agonist used to treat obesity and / or diabetes. In some such embodiments, the peptide drug is semaglutide, liraglutide, dulaglutide, abiglutide, or exenatide. Because the duration of action of the peptide drug is significantly prolonged when incorporated into the conjugates of this disclosure, GLP-1 agonist conjugates can reduce the frequency of administration, such as once weekly, once every two weeks, once every three weeks, or once monthly.

[0264] In some embodiments, the peptide drug is a glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic peptide (GIP) receptor agonist used to treat obesity and / or diabetes. In some such embodiments, the peptide drug is telposide. Because the duration of action of the peptide drug is significantly prolonged when incorporated into the conjugates of this disclosure, the GLP-1 / GIP agonist conjugates can reduce the frequency of administration, such as once weekly, once every two weeks, once every three weeks, or once monthly.

[0265] In some embodiments, the peptide is an endogenous opioid peptide. In some such embodiments, the peptide is endorphin-1 or endorphin-2. In other embodiments, the peptide is corticomerol.

[0266] In some embodiments of the conjugates of formula (V) or (V') or any variation thereof (e.g., formulas (Va) to (Vi)), D 1 It is an opioid antagonist. In some embodiments, the opioid antagonist is naloxone, nalmefen, natrozone, methylnaltrozone, nalbuphine, or nalprofen.

[0267] In some embodiments of the conjugates of formula (V) or (V') or any variation thereof (e.g., formulas (Va) to (Vi)), D 1 It is an opioid agonist. In some embodiments, the opioid agonist is morphine, oxycodone, hydromorphone, or methadone.

[0268] In some implementations, this document provides a conjugate of formula (VI'): (VI') Or its pharmaceutically acceptable salt, wherein: Each R 1 Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n is 0, 1, 2, or 3; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4 alkyl; L A C 3-20 Alkylene, wherein The C 3-20 Alkylene is optionally surrounded by one or more L 1a Replace, and One or more alkylene units are optionally selected from -O-, -NR y Substitution of - and -S- groups; Each L 1a Halogenated, C 1-6 Alkyl groups, or C groups substituted with halogen groups 1-6 alkyl; Each R y For H or C 1-4 alkyl; Y 1 For key or C 1-4 Alkylene; Y 2 For NR 6 , O or S; R 6 For H or C 1-4 alkyl; Each R 7 Independently selected from the following groups: hydrogen, halogen atoms (e.g., Cl, Br, or F), hydroxyl group, -OCF3, -NH2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 cycloalkyl; p is 0, 1, 2, or 3; and D 1 This refers to the pharmaceutical component.

[0269] In some implementations, this document provides a conjugate of formula (VI): (VI) Or its pharmaceutically acceptable salt, wherein: Each R 1 Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n is 0, 1, 2, or 3; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4 alkyl; L A C 3-20 Alkylene, wherein The C 3-20 Alkylene is optionally surrounded by one or more L 1a Replace, and One or more alkylene units are optionally selected from -O-, -NR ySubstitution of - and -S- groups; Each L 1a Halogenated, C 1-6 Alkyl groups, or C groups substituted with halogen groups 1-6 alkyl; Each R y For H or C 1-4 alkyl; Y 1 For key or C 1-4 Alkylene; Y 2 For NR 6 , O or S; R 6 For H or C 1-4 alkyl; Each R 7 Independently selected from the following groups: hydrogen, halogen atoms (e.g., Cl, Br, or F), hydroxyl group, -OCF3, -NH2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 cycloalkyl; p is 0, 1, 2, or 3; and D 1 This refers to the pharmaceutical component.

[0270] In some implementations, the formula (VI) conjugate has formula (VIa): (VIa) Or its pharmaceutically acceptable salt.

[0271] In some implementations, the formula (VI) conjugate has the formula (VIb): (VIb) Or its pharmaceutically acceptable salt.

[0272] In some implementations, the formula (VI) conjugate has the formula (VIc): (VIc) Or its pharmaceutically acceptable salt.

[0273] In some implementations, the formula (VI) conjugate has the formula (VId): (VId) Or its pharmaceutically acceptable salt.

[0274] In some implementations, the formula (VI) conjugate has the formula (VIe): (VIe) Or its pharmaceutically acceptable salt.

[0275] In some implementations, the formula (VI) conjugate has the formula (VIf): (VIf) Or its pharmaceutically acceptable salt.

[0276] In some implementations, the (VI') conjugate has the formula (VIg): (VIg) Or its pharmaceutically acceptable salt.

[0277] In some implementations, the formula (VI') conjugate has the formula (VIh): (VIh) Or its pharmaceutically acceptable salt.

[0278] In some embodiments of the conjugates of formula (VI) or (VI') or any variation thereof (e.g., formulas (VIa) to (VIh)), R 2 For H or C 1-2 Alkyl group. In some embodiments, R 2 It is H or -CH3. In some implementations, R 2 For H.

[0279] In some embodiments of the conjugates of formula (VI) or (VI') or any variation thereof (e.g., formulas (VIa) to (VIh)), R 3 For H or C 1-2 Alkyl group. In some embodiments, R 3 It is H or -CH3. In some implementations, R 3 For H.

[0280] In some embodiments of the conjugates of formula (VI) or (VI') or any variation thereof (e.g., formulas (VIa) to (VIh)), R 4 For H or C 1-2 Alkyl group. In some embodiments, R 4 It is H or -CH3. In some implementations, R 4 It is -CH3.

[0281] In some embodiments of the conjugates of formula (VI) or (VI') or any variation thereof (e.g., formulas (VIa) to (VIh)), R 5 For H or C 1-2 Alkyl group. In some embodiments, R 5 It is H or -CH3. In some implementations, R 5 It is -CH3.

[0282] In some embodiments of conjugates of formula (VI) or (VI') or any variation thereof (e.g., formulas (VIa) to (VIh)), n is 2 or 3. In some embodiments, n is 3.

[0283] In some embodiments of conjugates of formula (VI) or (VI') or any variation thereof (e.g., formulas (VIa) to (VIg)), m is 0, 1, or 2. In some embodiments, m is 0 or 1. In some embodiments, m is 0. In some embodiments, m is 1.

[0284] In some embodiments of conjugates of formula (VI) or (VI') or any variation thereof (e.g., formulas (VIa) to (VIg)), p is 0, 1, or 2. In some embodiments, p is 0 or 1. In some embodiments, p is 0. In some embodiments, p is 1.

[0285] In some embodiments of the conjugates of formula (VI) or (VI') or any variation thereof (e.g., formulas (VIa) to (VIh)), L A C 4-20 alkylene, wherein the C 4-20 Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally selected from -O-, -NR. x The - and -S- groups are substituted, where L 1a Halogenated, C 1-6 Alkyl groups, or C groups substituted with halogen groups 1-6 Alkyl group, wherein R x H or C optionally substituted with a halogen group 1-6 Alkyl group. In some embodiments, L A C 10-20 alkylene, wherein the C 10-20 Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally replaced by groups selected from -O-, -NH-, -N(CH3)-, and -S-. In some embodiments, L A C 10-15 alkylene, wherein the C 10-15 Alkylene is optionally surrounded by one or more L1a Substitution, and one or more alkylene units are optionally replaced by groups selected from -O-, -NH-, and -N(CH3)-. In some embodiments, L A C 5-10 alkylene, wherein the C 5-10 Alkylene is optionally surrounded by one or more L 1a The alkylene units are substituted, and one or more alkylene units are optionally replaced by groups selected from -O- and -NH-.

[0286] In some embodiments of the conjugates of formula (VI) or (VI') or any variation thereof (e.g., formulas (VIa) to (VIh)), L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part .

[0287] In some embodiments of the conjugates of formula (VI) or (VI') or any variation thereof (e.g., formulas (VIa) to (VIh)), D 1 For proteins. In some implementations, D 1The cytokine is used. In some embodiments, the cytokine is interleukin-2 (IL-2). In some embodiments, the cytokine is mutant IL-2, such as the cytokine described in US 9,206,943. In some embodiments, the cytokine is interleukin-15 (IL-15). In some embodiments, the cytokine is mutant IL-15. In one such embodiment, IL-15 is IL-15[N27D]. In some embodiments, the cytokine is interleukin-7 (IL-7). In some embodiments, the cytokine is interleukin-9 (IL-9). In some embodiments, the cytokine is interleukin-10 (IL-10). In some embodiments, the cytokine is interleukin-21 (IL-21). In some embodiments, the cytokine is interleukin-10 (IL-10).

[0288] In some embodiments of the conjugates of formula (VI) or (VI') or any variation thereof (e.g., formulas (VIa) to (VIh)), D 1 It is an anticancer agent. In some such implementations, D 1 For SN-38. In some such implementations, D 1 It is monomethylolpropionate E. In some such embodiments, D 1 For capecitabine. In some such implementations, D 1 For irinotecan. In some such implementations, D 1 It is trofluuridine. In some such embodiments, D 1 For Doxorubic stars. In some such implementations, D 1 For Dorcetathione. In some such implementations, D 1 It is Infiniti.

[0289] In some embodiments of the conjugates of formula (VI) or (VI') or any variation thereof (e.g., formulas (VIa) to (VIh)), D 1 It is a peptide.

[0290] In some embodiments, the peptide is RGK-MCA. In some embodiments, the peptide is neurotensin. In some embodiments, the peptide is gonadotropin-releasing hormone (GnRH). In some embodiments, the peptide is GIP (1-42) human peptide. In some embodiments, the peptide is D6-GnRH. In some embodiments, the peptide is insulin. In some embodiments, the peptide is teriparatide. In some embodiments, the peptide is salmon calcitonin. In some embodiments, the peptide is iparene. In some embodiments, the peptide is vasopressin. In some embodiments, the peptide is glucagon. In some embodiments, the peptide is alexiside. In some embodiments, the peptide is goserelin. In some embodiments, the peptide is leuprolide. In some embodiments, the peptide is teriparatide. In some embodiments, the peptide is lixinaside. In some embodiments, the peptide is desmopressin. In some embodiments, the peptide is oxytocin. In some embodiments, the peptide is corticotropin. In some embodiments, the peptide is sermorelin. In some embodiments, the peptide is bumenotide. In some embodiments, the peptide is abalotropic peptide. In some embodiments, the peptide is dilorelin.

[0291] In some embodiments, the peptide drug is a glucagon-like peptide-1 (GLP-1) receptor agonist used to treat obesity and / or diabetes. In some such embodiments, the peptide drug is semaglutide, liraglutide, dulaglutide, abiglutide, or exenatide. Because the duration of action of the peptide drug is significantly prolonged when incorporated into the conjugates of this disclosure, GLP-1 agonist conjugates can reduce the frequency of administration, such as once weekly, once every two weeks, once every three weeks, or once monthly.

[0292] In some embodiments, the peptide drug is a glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic peptide (GIP) receptor agonist used to treat obesity and / or diabetes. In some such embodiments, the peptide drug is telposide. Because the duration of action of the peptide drug is significantly prolonged when incorporated into the conjugates of this disclosure, the GLP-1 / GIP agonist conjugates can reduce the frequency of administration, such as once weekly, once every two weeks, once every three weeks, or once monthly.

[0293] In some embodiments, the peptide is an endogenous opioid peptide. In some such embodiments, the peptide is endorphin-1 or endorphin-2. In other embodiments, the peptide is corticomerol.

[0294] In some embodiments of the conjugates of formula (VI) or (VI') or any variation thereof (e.g., formulas (VIa) to (VIh)), D 1 This refers to the pharmaceutical component. In some implementations, the opioid antagonist is naloxone, nalmefen, natrozone, methylnaltrozone, nalbuphine, or nalprofen.

[0295] In some implementations, this document provides a conjugate of formula (VII'): (VII'), Or its pharmaceutically acceptable salt, wherein: Each R 1 Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n is 0, 1, 2, or 3; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4 alkyl; Y 1 For key or C 1-4 Alkylene; Y 2 For NR 6 , O or S; L B The click product is formed by the click reaction between the first click handle and the second click handle; R 6 For H or C 1-4 alkyl; Each R 7 Independently selected from the following groups: hydrogen, halogen atoms (e.g., Cl, Br, or F), hydroxyl group, -OCF3, -NH2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 cycloalkyl; p is 0, 1, 2 or 3; r is 1, 2, 3, 4, 5, 6, or 7; s is 1, 2, 3, 4, 5, 6, or 7; and D 1 This refers to the pharmaceutical component.

[0296] In some implementations, this document provides a conjugate of formula (VII): (VII), Or its pharmaceutically acceptable salt, wherein: Each R 1 Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n is 0, 1, 2, or 3; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4 alkyl; Y 1 For key or C 1-4 Alkylene; Y 2 For NR 6 , O or S; L B The click product is formed by the click reaction between the first click handle and the second click handle; R 6 For H or C 1-4 alkyl; Each R 7 Independently selected from the following groups: hydrogen, halogen atoms (e.g., Cl, Br, or F), hydroxyl group, -OCF3, -NH2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 cycloalkyl; p is 0, 1, 2 or 3; r is 1, 2, 3, 4, 5, 6, or 7; s is 1, 2, 3, 4, 5, 6, or 7; and D 1 This refers to the pharmaceutical component.

[0297] In some embodiments, the conjugate of formula (VII) has formula (VIIa): (VIIa) Or its pharmaceutically acceptable salt.

[0298] In some embodiments, the conjugate of formula (VII) has formula (VIIb): (VIIb) Or its pharmaceutically acceptable salt.

[0299] In some embodiments, the conjugate of formula (VII) has formula (VIIc): (VIIc) Or its pharmaceutically acceptable salt.

[0300] In some implementations, the conjugate of formula (VII') has formula (VIId): (VIId) Or its pharmaceutically acceptable salt.

[0301] In some embodiments of the conjugate of formula (VII) or (VII') or any variation thereof (e.g., formulas (VIIa) to (VIId)), R 2 For H or C 1-2 Alkyl group. In some embodiments, R 2 It is H or -CH3. In some implementations, R 2 For H.

[0302] In some embodiments of the conjugate of formula (VII) or (VII') or any variation thereof (e.g., formulas (VIIa) to (VIId)), R 3 For H or C 1-2 Alkyl group. In some embodiments, R 3 It is H or -CH3. In some implementations, R 3 For H.

[0303] In some embodiments of the conjugate of formula (VII) or (VII') or any variation thereof (e.g., formulas (VIIa) to (VIId)), R 4 For H or C1-2 Alkyl group. In some embodiments, R 4 It is H or -CH3. In some implementations, R 4 It is -CH3.

[0304] In some embodiments of the conjugate of formula (VII) or (VII') or any variation thereof (e.g., formulas (VIIa) to (VIId)), R 5 For H or C 1-2 Alkyl group. In some embodiments, R 5 It is H or -CH3. In some implementations, R 5 It is -CH3.

[0305] In some embodiments of conjugates of formula (VII) or (VII') or any variation thereof (e.g., formulas (VIIa) to (VIId)), n is 2 or 3. In some embodiments, n is 3.

[0306] In some embodiments of conjugates of formula (VII) or (VII') or any variation thereof (e.g., formulas (VIIa) to (VIId)), m is 0, 1, or 2. In some embodiments, m is 0 or 1. In some embodiments, m is 0. In some embodiments, m is 1.

[0307] In some embodiments of conjugates of formula (VII) or (VII') or any variation thereof (e.g., formulas (VIIa) to (VIId)), p is 0, 1, or 2. In some embodiments, p is 0 or 1. In some embodiments, p is 0. In some embodiments, p is 1.

[0308] In some embodiments of the conjugates of formula (VII) or (VII') or any variation thereof (e.g., formulas (VIIa) to (VIId)), the click product L B The triazine moiety. In some embodiments, the conjugate of any of formula (VII) or (VII'), or formulas (VIIa) to (VIId), comprises one of the following moieties: , where X 2 -CH2-, -NR x -, -S-, or -O-, where R x As defined elsewhere herein. In some embodiments, the conjugate comprises a portion In some implementations, the conjugate comprises a portion .

[0309] In some embodiments of the conjugate of formula (VII) or (VII') or any variation thereof (e.g., formulas (VIIa) to (VIId)), L A-L B Includes part or , where R 9 For H or C 1-4 Alkyl, R 10 For H or C 1-4 Alkyl group, and Q is -CH2-, -O-, -S-, -NH- or -N(C 1-4 alkyl)-.

[0310] In some embodiments of the conjugate of formula (VII) or (VII') and any variation thereof (e.g., formulas (VIIa) to (VIId)), D 1 For proteins. In some implementations, D 1 The cytokine is used. In some embodiments, the cytokine is interleukin-2 (IL-2). In some embodiments, the cytokine is mutant IL-2, such as the cytokine described in US 9,206,943. In some embodiments, the cytokine is interleukin-15 (IL-15). In some embodiments, the cytokine is mutant IL-15. In one such embodiment, IL-15 is IL-15[N27D]. In some embodiments, the cytokine is interleukin-7 (IL-7). In some embodiments, the cytokine is interleukin-9 (IL-9). In some embodiments, the cytokine is interleukin-10 (IL-10). In some embodiments, the cytokine is interleukin-21 (IL-21). In some embodiments, the cytokine is interleukin-10 (IL-10).

[0311] In some embodiments of the conjugate of formula (VII) or (VII') or any variation thereof (e.g., formulas (VIIa) to (VIId)), D 1 It is an anticancer agent. In some such implementations, D 1 For SN-38. In some such implementations, D 1 It is monomethylolpropionate E. In some such embodiments, D 1 For capecitabine. In some such implementations, D 1 For irinotecan. In some such implementations, D 1 It is trofluuridine. In some such embodiments, D 1 For Doxorubic stars. In some such implementations, D 1 For Dorcetathione. In some such implementations, D 1 It is Infiniti.

[0312] In some embodiments of the conjugate of formula (VII) or (VII') or any variation thereof (e.g., formulas (VIIa) to (VIId)), D 1 It is a peptide.

[0313] In some embodiments, the peptide is RGK-MCA. In some embodiments, the peptide is neurotensin. In some embodiments, the peptide is gonadotropin-releasing hormone (GnRH). In some embodiments, the peptide is GIP (1-42) human peptide. In some embodiments, the peptide is D6-GnRH. In some embodiments, the peptide is insulin. In some embodiments, the peptide is teriparatide. In some embodiments, the peptide is salmon calcitonin. In some embodiments, the peptide is iparene. In some embodiments, the peptide is vasopressin. In some embodiments, the peptide is glucagon. In some embodiments, the peptide is alexiside. In some embodiments, the peptide is goserelin. In some embodiments, the peptide is leuprolide. In some embodiments, the peptide is teriparatide. In some embodiments, the peptide is lixinaside. In some embodiments, the peptide is desmopressin. In some embodiments, the peptide is oxytocin. In some embodiments, the peptide is corticotropin. In some embodiments, the peptide is sermorelin. In some embodiments, the peptide is bumenotide. In some embodiments, the peptide is abalotropic peptide. In some embodiments, the peptide is dilorelin.

[0314] In some embodiments, the peptide drug is a glucagon-like peptide-1 (GLP-1) receptor agonist used to treat obesity and / or diabetes. In some such embodiments, the peptide drug is semaglutide, liraglutide, dulaglutide, abiglutide, or exenatide. Because the duration of action of the peptide drug is significantly prolonged when incorporated into the conjugates of this disclosure, GLP-1 agonist conjugates can reduce the frequency of administration, such as once weekly, once every two weeks, once every three weeks, or once monthly.

[0315] In some embodiments, the peptide drug is a glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic peptide (GIP) receptor agonist used to treat obesity and / or diabetes. In some such embodiments, the peptide drug is telposide. Because the duration of action of the peptide drug is significantly prolonged when incorporated into the conjugates of this disclosure, the GLP-1 / GIP agonist conjugates can reduce the frequency of administration, such as once weekly, once every two weeks, once every three weeks, or once monthly.

[0316] In some embodiments, the peptide is an endogenous opioid peptide. In some such embodiments, the peptide is endorphin-1 or endorphin-2. In other embodiments, the peptide is corticomerol.

[0317] In some embodiments of the conjugate of formula (VII) or (VII') or any variation thereof (e.g., formulas (VIIa) to (VIId)), D 1 It is an opioid antagonist. In some embodiments, the opioid antagonist is naloxone, nalmefen, natrozone, methylnaltrozone, nalbuphine, or nalprofen.

[0318] In some embodiments of the conjugate of formula (VII) or (VII') or any variation thereof (e.g., formulas (VIIa) to (VIId)), D 1 It is an opioid agonist. In some embodiments, the opioid agonist is morphine, oxycodone, hydromorphone, or methadone.

[0319] In some implementations, this document provides a conjugate of formula (VIII'): (VIII') Or its pharmaceutically acceptable salt, wherein: Each R 1 Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n is 0, 1, 2, or 3; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4 alkyl; Y 1 For key or C 1-4 Alkylene; Y 2 For NR 6 , O or S; L B The click product is formed by the click reaction between the first click handle and the second click handle; R 6 For H or C 1-4 alkyl; Each R 7Independently selected from the following groups: hydrogen, halogen atoms (e.g., Cl, Br, or F), hydroxyl group, -OCF3, -NH2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 cycloalkyl; p is 0, 1, 2 or 3; r is 1, 2, 3, 4, 5, 6, or 7; s can be 1, 2, 3, 4, 5, 6, or 7; t is 1, 2, 3, or 4; u is 1, 2, 3, or 4; and D 1 This refers to the pharmaceutical component.

[0320] In some implementations, this document provides a conjugate of formula (VIII): (VIII) Or its pharmaceutically acceptable salt, wherein: Each R 1 Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n is 0, 1, 2, or 3; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4 alkyl; Y 1 For key or C 1-4 Alkylene; Y 2 For NR 6 , O or S; L BThe click product is formed by the click reaction between the first click handle and the second click handle; R 6 For H or C 1-4 alkyl; Each R 7 Independently selected from the following groups: hydrogen, halogen atoms (e.g., Cl, Br, or F), hydroxyl group, -OCF3, -NH2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 cycloalkyl; p is 0, 1, 2 or 3; r is 1, 2, 3, 4, 5, 6, or 7; s can be 1, 2, 3, 4, 5, 6, or 7; t is 1, 2, 3, or 4; u is 1, 2, 3, or 4; and D 1 This refers to the pharmaceutical component.

[0321] In some embodiments, the (VIII) conjugate has the formula (VIIIa): (VIIIa) Or its pharmaceutically acceptable salt.

[0322] In some embodiments, the (VIII) conjugate has the formula (VIIIb): (VIIIb) Or its pharmaceutically acceptable salt.

[0323] In some embodiments, the (VIII) conjugate has the formula (VIIIc): (VIIIc) Or its pharmaceutically acceptable salt.

[0324] In some embodiments, the (VIII) conjugate has the formula (VIIId): (VIIId) Or its pharmaceutically acceptable salt.

[0325] In some embodiments, the (VIII) conjugate has the formula (VIIIe): (VIIIe) Or its pharmaceutically acceptable salt.

[0326] In some implementations, the (VIII') conjugate has the formula (VIIIf): (VIIIf) Or its pharmaceutically acceptable salt.

[0327] In some embodiments of the conjugates of formula (VIII) or (VIII') or any variation thereof (e.g., formulas (VIIIa) to (VIIIf)), R 2 For H or C 1-2 Alkyl group. In some embodiments, R 2 It is H or -CH3. In some implementations, R 2 For H.

[0328] In some embodiments of the conjugates of formula (VIII) or (VIII') or any variation thereof (e.g., formulas (VIIIa) to (VIIIf)), R 3 For H or C 1-2 Alkyl group. In some embodiments, R 3 It is H or -CH3. In some implementations, R 3 For H.

[0329] In some embodiments of the conjugates of formula (VIII) or (VIII') or any variation thereof (e.g., formulas (VIIIa) to (VIIIf)), R 4 For H or C 1-2 Alkyl group. In some embodiments, R 4 It is H or -CH3. In some implementations, R 4 It is -CH3.

[0330] In some embodiments of the conjugates of formula (VIII) or (VIII') or any variation thereof (e.g., formulas (VIIIa) to (VIIIf)), R 5 For H or C 1-2 Alkyl group. In some embodiments, R 5 It is H or -CH3. In some implementations, R 5 It is -CH3.

[0331] In some embodiments of conjugates of formula (VIII) or (VIII') or any variation thereof (e.g., formulas (VIIIa) to (VIIIf)), n is 2 or 3. In some embodiments, n is 3.

[0332] In some embodiments of conjugates of formula (VIII) or (VIII') or any variation thereof (e.g., formulas (VIIIa) to (VIIIf)), m is 0, 1, or 2. In some embodiments, m is 0 or 1. In some embodiments, m is 0. In some embodiments, m is 1.

[0333] In some embodiments of conjugates of formula (VIII) or (VIII') or any variation thereof (e.g., formula (VIIIa) or formulas (VIIIa) through (VIIIf)), p is 0, 1, or 2. In some embodiments, p is 0 or 1. In some embodiments, p is 0. In some embodiments, p is 1.

[0334] In some embodiments of the conjugates of formula (VIII) or (VIII') or any variation thereof (e.g., formulas (VIIIa) to (VIIIb)), the click product L B The triazine moiety. In some embodiments, the conjugate of any one of formulas (VIII), (VIIIa) to (VIIIb) comprises one of the following moieties: , where X 2 -CH2-, -NR x -, -S-, or -O-, where R x As defined elsewhere herein. In some embodiments, the conjugate comprises a portion In some implementations, the conjugate comprises a portion .

[0335] In some embodiments of the conjugates of formula (VIII) or (VIII') or any variation thereof (e.g., formulas (VIIIa) to (VIIIb)), L A -L B Includes part or , where R 9 For H or C 1-4 Alkyl, R 10 For H or C 1-4 Alkyl group, and Q is -CH2-, -O-, -S-, -NH- or -N(C 1-4 alkyl)-.

[0336] In some embodiments of the conjugates of formula (VIII) or (VIII') or any variation thereof (e.g., formulas (VIIIa) to (VIIIf)), D1 For proteins. In some implementations, D 1 The cytokine is used. In some embodiments, the cytokine is interleukin-2 (IL-2). In some embodiments, the cytokine is mutant IL-2, such as the cytokine described in US 9,206,943. In some embodiments, the cytokine is interleukin-15 (IL-15). In some embodiments, the cytokine is mutant IL-15. In one such embodiment, IL-15 is IL-15[N27D]. In some embodiments, the cytokine is interleukin-7 (IL-7). In some embodiments, the cytokine is interleukin-9 (IL-9). In some embodiments, the cytokine is interleukin-10 (IL-10). In some embodiments, the cytokine is interleukin-21 (IL-21). In some embodiments, the cytokine is interleukin-10 (IL-10).

[0337] In some embodiments of the conjugates of formula (VIII) or (VIII') or any variation thereof (e.g., formulas (VIIIa) to (VIIIf)), D 1 It is an anticancer agent. In some such implementations, D 1 For SN-38. In some such implementations, D 1 It is monomethylolpropionate E. In some such embodiments, D 1 For capecitabine. In some such implementations, D 1 For irinotecan. In some such implementations, D 1 It is trofluuridine. In some such embodiments, D 1 For Doxorubic stars. In some such implementations, D 1 For Dorcetathione. In some such implementations, D 1 It is Infiniti.

[0338] In some embodiments of the conjugates of formula (VIII) or (VIII') or any variation thereof (e.g., formulas (VIIIa) to (VIIIf)), D 1 It is a peptide.

[0339] In some embodiments, the peptide is RGK-MCA. In some embodiments, the peptide is neurotensin. In some embodiments, the peptide is gonadotropin-releasing hormone (GnRH). In some embodiments, the peptide is GIP (1-42) human peptide. In some embodiments, the peptide is D6-GnRH. In some embodiments, the peptide is insulin. In some embodiments, the peptide is teriparatide. In some embodiments, the peptide is salmon calcitonin. In some embodiments, the peptide is iparene. In some embodiments, the peptide is vasopressin. In some embodiments, the peptide is glucagon. In some embodiments, the peptide is alexiside. In some embodiments, the peptide is goserelin. In some embodiments, the peptide is leuprolide. In some embodiments, the peptide is teriparatide. In some embodiments, the peptide is lixinaside. In some embodiments, the peptide is desmopressin. In some embodiments, the peptide is oxytocin. In some embodiments, the peptide is corticotropin. In some embodiments, the peptide is sermorelin. In some embodiments, the peptide is bumenotide. In some embodiments, the peptide is abalotropic peptide. In some embodiments, the peptide is dilorelin.

[0340] In some embodiments, the peptide drug is a glucagon-like peptide-1 (GLP-1) receptor agonist used to treat obesity and / or diabetes. In some such embodiments, the peptide drug is semaglutide, liraglutide, dulaglutide, abiglutide, or exenatide. Because the duration of action of the peptide drug is significantly prolonged when incorporated into the conjugates of this disclosure, GLP-1 agonist conjugates can reduce the frequency of administration, such as once weekly, once every two weeks, once every three weeks, or once monthly.

[0341] In some embodiments, the peptide drug is a glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic peptide (GIP) receptor agonist used to treat obesity and / or diabetes. In some such embodiments, the peptide drug is telposide. Because the duration of action of the peptide drug is significantly prolonged when incorporated into the conjugates of this disclosure, the GLP-1 / GIP agonist conjugates can reduce the frequency of administration, such as once weekly, once every two weeks, once every three weeks, or once monthly.

[0342] In some embodiments, the peptide is an endogenous opioid peptide. In some such embodiments, the peptide is endorphin-1 or endorphin-2. In other embodiments, the peptide is corticomerol.

[0343] In some embodiments of the conjugates of formula (VIII) or (VIII') or any variation thereof (e.g., formulas (VIIIa) to (VIIIf)), D 1It is an opioid antagonist. In some embodiments, the opioid antagonist is naloxone, nalmefen, natrozone, methylnaltrozone, nalbuphine, or nalprofen.

[0344] In some embodiments of the conjugates of formula (VIII) or (VIII') or any variation thereof (e.g., formulas (VIIIa) to (VIIIf)), D 1 It is an opioid agonist. In some embodiments, the opioid agonist is morphine, oxycodone, hydromorphone, or methadone.

[0345] In some implementations, this document provides a conjugate of formula (IX): (IX), Or its pharmaceutically acceptable salt, wherein: Each R 1 Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n is 0, 1, 2, or 3; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4 alkyl; L stands for connector; Z 1 C 1-6 Alkyl, C 6-10 Aryl, or 5 to 10-membered heteroaryl, among which The C 1-6 Alkyl groups are optionally surrounded by one or more Z 1a Replace, and The C 6-10 Aryl or 5- to 10-membered heteroaryl groups are optionally surrounded by one or more Z groups. 1b replace; Each Z 1a or Z 1b Independently halogenated, C 1-6 alkoxy or C-group optionally substituted with a halogen group 1-6 Alkyl; and D 1 This refers to the pharmaceutical component.

[0346] In some embodiments of the (IX) conjugate, R 2 For H or C 1-2 Alkyl group. In some embodiments, R 2 It is H or -CH3. In some implementations, R 2 For H.

[0347] In some embodiments of the (IX) conjugate, R 3 For H or C 1-2 Alkyl group. In some embodiments, R 3 It is H or -CH3. In some implementations, R 3 For H.

[0348] In some embodiments of the (IX) conjugate, R 4 For H or C 1-2 Alkyl group. In some embodiments, R 4 It is H or -CH3. In some implementations, R 4 It is -CH3.

[0349] In some embodiments of the formula (IX) conjugate, R 5 For H or C 1-2 Alkyl group. In some embodiments, R 5 It is H or -CH3. In some implementations, R 5 It is -CH3.

[0350] In some embodiments of the conjugate of formula (IX), n is 2 or 3. In some embodiments, n is 3.

[0351] In some embodiments of the conjugate of formula (IX), m is 0, 1, or 2. In some embodiments, m is 0 or 1. In some embodiments, m is 0. In some embodiments, m is 1.

[0352] In some embodiments of the (IX) conjugate, L is C 4-20 alkylene, wherein the C 4-20 Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally selected from -O-, -NR. x The - and -S- groups are substituted, where L 1a Halogenated, C 1-6 Alkyl groups, or C groups substituted with halogen groups 1-6 Alkyl group. In some embodiments, L is C. 10-20 alkylene, wherein the C 10-20Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally replaced by groups selected from -O-, -NH-, -N(CH3)-, and -S-. In some embodiments, L is C 10-15 alkylene, wherein the C 10-15 Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally replaced by groups selected from -O-, -NH-, and -N(CH3)-. In some embodiments, L is C 5-10 alkylene, wherein the C 5-10 Alkylene is optionally surrounded by one or more L 1a The alkylene units are substituted, and one or more alkylene units are optionally replaced by groups selected from -O- and -NH-.

[0353] In some embodiments of the formula (IX) conjugate, L contains a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion In some implementations, L includes a portion .

[0354] In some embodiments of conjugates of formula (IX) or any variation thereof, D 1 For proteins. In some implementations, D 1The cytokine is used. In some embodiments, the cytokine is interleukin-2 (IL-2). In some embodiments, the cytokine is mutant IL-2, such as the cytokine described in US 9,206,943. In some embodiments, the cytokine is interleukin-15 (IL-15). In some embodiments, the cytokine is mutant IL-15. In one such embodiment, IL-15 is IL-15[N27D]. In some embodiments, the cytokine is interleukin-7 (IL-7). In some embodiments, the cytokine is interleukin-9 (IL-9). In some embodiments, the cytokine is interleukin-10 (IL-10). In some embodiments, the cytokine is interleukin-21 (IL-21). In some embodiments, the cytokine is interleukin-10 (IL-10).

[0355] In some embodiments of conjugates of formula (IX) or any variation thereof, D 1 It is an anticancer agent. In some such implementations, D 1 For SN-38. In some such implementations, D 1 It is monomethylolpropionate E. In some such embodiments, D 1 For capecitabine. In some such implementations, D 1 For irinotecan. In some such implementations, D 1 It is trofluuridine. In some such embodiments, D 1 For Doxorubic stars. In some such implementations, D 1 For Dorcetathione. In some such implementations, D 1 It is Infiniti.

[0356] In some embodiments of conjugates of formula (IX) or any variation thereof, D 1 It is a peptide.

[0357] In some embodiments, the peptide is RGK-MCA. In some embodiments, the peptide is neurotensin. In some embodiments, the peptide is gonadotropin-releasing hormone (GnRH). In some embodiments, the peptide is GIP (1-42) human peptide. In some embodiments, the peptide is D6-GnRH. In some embodiments, the peptide is insulin. In some embodiments, the peptide is teriparatide. In some embodiments, the peptide is salmon calcitonin. In some embodiments, the peptide is iparene. In some embodiments, the peptide is vasopressin. In some embodiments, the peptide is glucagon. In some embodiments, the peptide is alexiside. In some embodiments, the peptide is goserelin. In some embodiments, the peptide is leuprolide. In some embodiments, the peptide is teriparatide. In some embodiments, the peptide is lixinaside. In some embodiments, the peptide is desmopressin. In some embodiments, the peptide is oxytocin. In some embodiments, the peptide is corticotropin. In some embodiments, the peptide is sermorelin. In some embodiments, the peptide is bumenotide. In some embodiments, the peptide is abalotropic peptide. In some embodiments, the peptide is dilorelin.

[0358] In some embodiments, the peptide drug is a glucagon-like peptide-1 (GLP-1) receptor agonist used to treat obesity and / or diabetes. In some such embodiments, the peptide drug is semaglutide, liraglutide, dulaglutide, abiglutide, or exenatide. Because the duration of action of the peptide drug is significantly prolonged when incorporated into the conjugates of this disclosure, GLP-1 agonist conjugates can reduce the frequency of administration, such as once weekly, once every two weeks, once every three weeks, or once monthly.

[0359] In some embodiments, the peptide drug is a glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic peptide (GIP) receptor agonist used to treat obesity and / or diabetes. In some such embodiments, the peptide drug is telposide. Because the duration of action of the peptide drug is significantly prolonged when incorporated into the conjugates of this disclosure, the GLP-1 / GIP agonist conjugates can reduce the frequency of administration, such as once weekly, once every two weeks, once every three weeks, or once monthly.

[0360] In some embodiments, the peptide is an endogenous opioid peptide. In some such embodiments, the peptide is endorphin-1 or endorphin-2. In other embodiments, the peptide is corticomerol.

[0361] In some embodiments of conjugates of formula (IX) or any variation thereof, D 1 It is an opioid antagonist. In some embodiments, the opioid antagonist is naloxone, nalmefen, natrozone, methylnaltrozone, nalbuphine, or nalprofen.

[0362] In some embodiments of conjugates of formula (IX) or any variation thereof, D 1 It is an opioid agonist. In some embodiments, the opioid agonist is morphine, oxycodone, hydromorphone, or methadone.

[0363] In some implementations, this document provides a conjugate of formula (X): (X), Or its pharmaceutically acceptable salt, wherein: Each R 1 Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n is 0, 1, 2, or 3; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4 alkyl; L A C 3-20 Alkylene, wherein The C 3-20 Alkylene is optionally surrounded by one or more L 1a Replace, and One or more alkylene units are optionally selected from -O-, -NR y Substitution of - and -S- groups; Each L 1a Halogenated, C 1-6 Alkyl groups, or C groups substituted with halogen groups 1-6 alkyl; Each R y For H or C 1-4 alkyl; L B The click product is formed by the click reaction between the first click handle and the second click handle; Z 1 C 1-6 Alkyl, C 6-10 Aryl, or 5 to 10-membered heteroaryl, among which The C 1-6 Alkyl groups are optionally surrounded by one or more Z 1a Replace, and The C 6-10 Aryl or 5- to 10-membered heteroaryl groups are optionally surrounded by one or more Z groups. 1b replace; Each Z 1a or Z 1b Independently halogenated, C 1-6 alkoxy or C-group optionally substituted with a halogen group 1-6 Alkyl; and D 1 This refers to the pharmaceutical component.

[0364] In some embodiments of the (X) conjugate, R 2 For H or C 1-2 Alkyl group. In some embodiments, R 2 It is H or -CH3. In some implementations, R 2 For H.

[0365] In some embodiments of the (X) conjugate, R 3 For H or C 1-2 Alkyl group. In some embodiments, R 3 It is H or -CH3. In some implementations, R 3 For H.

[0366] In some embodiments of the (X) conjugate, R 4 For H or C 1-2 Alkyl group. In some embodiments, R 4 It is H or -CH3. In some implementations, R 4 It is -CH3.

[0367] In some embodiments of the (X) conjugate, R 5 For H or C 1-2 Alkyl group. In some embodiments, R 5 It is H or -CH3. In some implementations, R 5 It is -CH3.

[0368] In some embodiments of the (X) conjugate, n is 2 or 3. In some embodiments, n is 3.

[0369] In some embodiments of the (X) conjugate, m is 0, 1, or 2. In some embodiments, m is 0 or 1. In some embodiments, m is 0. In some embodiments, m is 1.

[0370] In some embodiments of the (X) conjugate, L A C4-20 Alkylene, wherein C 4-20 Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally selected from -O-, -NR. x The - and -S- groups are substituted, where L 1a Halogenated, C 1-6 Alkyl groups, or C groups substituted with halogen groups 1-6 Alkyl group, wherein R x H or C optionally substituted with a halogen group 1-6 Alkyl group. In some embodiments, L A C 10-20 alkylene, wherein the C 10-20 Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally replaced by groups selected from -O-, -NH-, -N(CH3)-, and -S-. In some embodiments, L A C 10-15 alkylene, wherein the C 10-15 Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally replaced by groups selected from -O-, -NH-, and -N(CH3)-. In some embodiments, L A C 5-10 alkylene, wherein the C 5-10 Alkylene is optionally surrounded by one or more L 1a The alkylene units are substituted, and one or more alkylene units are optionally replaced by groups selected from -O- and -NH-.

[0371] In some embodiments of the (X) conjugate, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part .

[0372] In some embodiments of the formula (X) conjugate, the click product L B The triazine moiety. In some embodiments, the (X) conjugate comprises one of the following moieties: , where X 2 -CH2-, -NR x -, -S-, or -O-, where R x As defined elsewhere herein. In some embodiments, the conjugate comprises a portion In some implementations, the conjugate comprises a portion .

[0373] In some embodiments of the (X) conjugate, L A -L B Includes part or , where R 9 For H or C 1-4 Alkyl, R 10 For H or C 1-4 Alkyl group, and Q is -CH2-, -O-, -S-, -NH- or -N(C 1-4 alkyl)-.

[0374] In some embodiments of conjugates of formula (X) or any variation thereof, D 1 For proteins. In some implementations, D 1The cytokine is used. In some embodiments, the cytokine is interleukin-2 (IL-2). In some embodiments, the cytokine is mutant IL-2, such as the cytokine described in US 9,206,943. In some embodiments, the cytokine is interleukin-15 (IL-15). In some embodiments, the cytokine is mutant IL-15. In one such embodiment, IL-15 is IL-15[N27D]. In some embodiments, the cytokine is interleukin-7 (IL-7). In some embodiments, the cytokine is interleukin-9 (IL-9). In some embodiments, the cytokine is interleukin-10 (IL-10). In some embodiments, the cytokine is interleukin-21 (IL-21). In some embodiments, the cytokine is interleukin-10 (IL-10).

[0375] In some embodiments of conjugates of formula (X) or any variation thereof, D 1 It is an anticancer agent. In some such implementations, D 1 For SN-38. In some such implementations, D 1 It is monomethylolpropionate E. In some such embodiments, D 1 For capecitabine. In some such implementations, D 1 For irinotecan. In some such implementations, D 1 It is trofluuridine. In some such embodiments, D 1 For Doxorubic stars. In some such implementations, D 1 For Dorcetathione. In some such implementations, D 1 It is Infiniti.

[0376] In some embodiments of conjugates of formula (X) or any variation thereof, D 1 It is a peptide.

[0377] In some embodiments, the peptide is RGK-MCA. In some embodiments, the peptide is neurotensin. In some embodiments, the peptide is gonadotropin-releasing hormone (GnRH). In some embodiments, the peptide is GIP (1-42) human peptide. In some embodiments, the peptide is D6-GnRH. In some embodiments, the peptide is insulin. In some embodiments, the peptide is teriparatide. In some embodiments, the peptide is salmon calcitonin. In some embodiments, the peptide is iparene. In some embodiments, the peptide is vasopressin. In some embodiments, the peptide is glucagon. In some embodiments, the peptide is alexiside. In some embodiments, the peptide is goserelin. In some embodiments, the peptide is leuprolide. In some embodiments, the peptide is teriparatide. In some embodiments, the peptide is lixinaside. In some embodiments, the peptide is desmopressin. In some embodiments, the peptide is oxytocin. In some embodiments, the peptide is corticotropin. In some embodiments, the peptide is sermorelin. In some embodiments, the peptide is bumenotide. In some embodiments, the peptide is abalotropic peptide. In some embodiments, the peptide is dilorelin.

[0378] In some embodiments, the peptide drug is a glucagon-like peptide-1 (GLP-1) receptor agonist used to treat obesity and / or diabetes. In some such embodiments, the peptide drug is semaglutide, liraglutide, dulaglutide, abiglutide, or exenatide. Because the duration of action of the peptide drug is significantly prolonged when incorporated into the conjugates of this disclosure, GLP-1 agonist conjugates can reduce the frequency of administration, such as once weekly, once every two weeks, once every three weeks, or once monthly.

[0379] In some embodiments, the peptide drug is a glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic peptide (GIP) receptor agonist used to treat obesity and / or diabetes. In some such embodiments, the peptide drug is telposide. Because the duration of action of the peptide drug is significantly prolonged when incorporated into the conjugates of this disclosure, the GLP-1 / GIP agonist conjugates can reduce the frequency of administration, such as once weekly, once every two weeks, once every three weeks, or once monthly.

[0380] In some embodiments, the peptide is an endogenous opioid peptide. In some such embodiments, the peptide is endorphin-1 or endorphin-2. In other embodiments, the peptide is corticomerol.

[0381] In some embodiments of conjugates of formula (X) or any variation thereof, D 1 It is an opioid antagonist. In some embodiments, the opioid antagonist is naloxone, nalmefen, natrozone, methylnaltrozone, nalbuphine, or nalprofen.

[0382] In some embodiments of conjugates of formula (X) or any variation thereof, D 1 It is an opioid agonist. In some embodiments, the opioid agonist is morphine, oxycodone, hydromorphone, or methadone.

[0383] In some implementations, this document provides a conjugate of formula (XI): (XI), Or its pharmaceutically acceptable salt, wherein: Each R 1 Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n is 0, 1, 2, or 3; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4 alkyl; L A C 3-20 Alkylene, wherein The C 3-20 Alkylene is optionally surrounded by one or more L 1a Replace, and One or more alkylene units are optionally selected from -O-, -NR y Substitution of - and -S- groups; Each L 1a Halogenated, C 1-6 Alkyl groups, or C groups substituted with halogen groups 1-6 alkyl; Each R y For H or C 1-4 alkyl; Z 1 C 1-6 Alkyl, C 6-10 Aryl, or 5 to 10-membered heteroaryl, among which The C 1-6 Alkyl groups are optionally surrounded by one or more Z 1a Replace, and The C 6-10 Aryl or 5- to 10-membered heteroaryl groups are optionally surrounded by one or more Z groups. 1b replace; Each Z 1a or Z 1b Independently halogenated, C 1-6 alkoxy or C-group optionally substituted with a halogen group 1-6 Alkyl; and D 1 This refers to the pharmaceutical component.

[0384] In some embodiments of the conjugate of formula (XI), R 2 For H or C 1-2 Alkyl group. In some embodiments, R 2 It is H or -CH3. In some implementations, R 2 For H.

[0385] In some embodiments of the conjugate of formula (XI), R 3 For H or C 1-2 Alkyl group. In some embodiments, R 3 It is H or -CH3. In some implementations, R 3 For H.

[0386] In some embodiments of the conjugate of formula (XI), R 4 For H or C 1-2 Alkyl group. In some embodiments, R 4 It is H or -CH3. In some implementations, R 4 It is -CH3.

[0387] In some embodiments of the conjugate of formula (XI), R 5 For H or C 1-2 Alkyl group. In some embodiments, R 5 It is H or -CH3. In some implementations, R 5 It is -CH3.

[0388] In some embodiments of the conjugate of formula (XI), n is 2 or 3. In some embodiments, n is 3.

[0389] In some embodiments of the conjugate of formula (XI), m is 0, 1, or 2. In some embodiments, m is 0 or 1. In some embodiments, m is 0. In some embodiments, m is 1.

[0390] In some embodiments of the conjugate of formula (XI), L A C 4-20 Alkylene, wherein C 4-20Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally selected from -O-, -NR. x The - and -S- groups are substituted, where L 1a Halogenated, C 1-6 Alkyl groups, or C groups substituted with halogen groups 1-6 Alkyl group, wherein R x H or C optionally substituted with a halogen group 1-6 Alkyl group. In some embodiments, L A C 10-20 alkylene, wherein the C 10-20 Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally replaced by groups selected from -O-, -NH-, -N(CH3)-, and -S-. In some embodiments, L A C 10-15 alkylene, wherein the C 10-15 Alkylene is optionally surrounded by one or more L 1a Substitution, and one or more alkylene units are optionally replaced by groups selected from -O-, -NH-, and -N(CH3)-. In some embodiments, L A C 5-10 alkylene, wherein the C 5-10 Alkylene is optionally surrounded by one or more L 1a The alkylene units are substituted, and one or more alkylene units are optionally replaced by groups selected from -O- and -NH-.

[0391] In some embodiments of the conjugate of formula (XI), L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L AIncludes part In some implementations, L A Includes part In some implementations, L A Includes part In some implementations, L A Includes part .

[0392] In some embodiments of conjugates of formula (XI) or any variation thereof, D 1 For proteins. In some implementations, D 1 The cytokine is used. In some embodiments, the cytokine is interleukin-2 (IL-2). In some embodiments, the cytokine is mutant IL-2, such as the cytokine described in US 9,206,943. In some embodiments, the cytokine is interleukin-15 (IL-15). In some embodiments, the cytokine is mutant IL-15. In one such embodiment, IL-15 is IL-15[N27D]. In some embodiments, the cytokine is interleukin-7 (IL-7). In some embodiments, the cytokine is interleukin-9 (IL-9). In some embodiments, the cytokine is interleukin-10 (IL-10). In some embodiments, the cytokine is interleukin-21 (IL-21). In some embodiments, the cytokine is interleukin-10 (IL-10).

[0393] In some embodiments of conjugates of formula (XI) or any variation thereof, D 1 It is an anticancer agent. In some such implementations, D 1 For SN-38. In some such implementations, D 1 It is monomethylolpropionate E. In some such embodiments, D 1 For capecitabine. In some such implementations, D 1 For irinotecan. In some such implementations, D 1 It is trofluuridine. In some such embodiments, D 1 For Doxorubic stars. In some such implementations, D 1 For Dorcetathione. In some such implementations, D 1 It is Infiniti.

[0394] In some embodiments of conjugates of formula (XI) or any variation thereof, D 1 It is a peptide.

[0395] In some embodiments, the peptide is RGK-MCA. In some embodiments, the peptide is neurotensin. In some embodiments, the peptide is gonadotropin-releasing hormone (GnRH). In some embodiments, the peptide is GIP (1-42) human peptide. In some embodiments, the peptide is D6-GnRH. In some embodiments, the peptide is insulin. In some embodiments, the peptide is teriparatide. In some embodiments, the peptide is salmon calcitonin. In some embodiments, the peptide is iparene. In some embodiments, the peptide is vasopressin. In some embodiments, the peptide is glucagon. In some embodiments, the peptide is alexiside. In some embodiments, the peptide is goserelin. In some embodiments, the peptide is leuprolide. In some embodiments, the peptide is teriparatide. In some embodiments, the peptide is lixinaside. In some embodiments, the peptide is desmopressin. In some embodiments, the peptide is oxytocin. In some embodiments, the peptide is corticotropin. In some embodiments, the peptide is sermorelin. In some embodiments, the peptide is bumenotide. In some embodiments, the peptide is abalotropic peptide. In some embodiments, the peptide is dilorelin.

[0396] In some embodiments, the peptide drug is a glucagon-like peptide-1 (GLP-1) receptor agonist used to treat obesity and / or diabetes. In some such embodiments, the peptide drug is semaglutide, liraglutide, dulaglutide, abiglutide, or exenatide. Because the duration of action of the peptide drug is significantly prolonged when incorporated into the conjugates of this disclosure, GLP-1 agonist conjugates can reduce the frequency of administration, such as once weekly, once every two weeks, once every three weeks, or once monthly.

[0397] In some embodiments, the peptide drug is a glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic peptide (GIP) receptor agonist used to treat obesity and / or diabetes. In some such embodiments, the peptide drug is telposide. Because the duration of action of the peptide drug is significantly prolonged when incorporated into the conjugates of this disclosure, the GLP-1 / GIP agonist conjugates can reduce the frequency of administration, such as once weekly, once every two weeks, once every three weeks, or once monthly.

[0398] In some embodiments, the peptide is an endogenous opioid peptide. In some such embodiments, the peptide is endorphin-1 or endorphin-2. In other embodiments, the peptide is corticomerol.

[0399] In some embodiments of conjugates of formula (XI) or any variation thereof, D 1 It is an opioid antagonist. In some embodiments, the opioid antagonist is naloxone, nalmefen, natrozone, methylnaltrozone, nalbuphine, or nalprofen.

[0400] In some embodiments of conjugates of formula (XI) or any variation thereof, D 1 It is an opioid agonist. In some embodiments, the opioid agonist is morphine, oxycodone, hydromorphone, or methadone.

[0401] In some embodiments, this document provides a conjugate having the following structure: (XII), Where D 1 This refers to the pharmaceutical component.

[0402] In some embodiments of the (XII) conjugate, D 1 For proteins. In some implementations, D 1 The cytokine is used. In some embodiments, the cytokine is interleukin-2 (IL-2). In some embodiments, the cytokine is mutant IL-2, such as the cytokine described in US 9,206,943. In some embodiments, the cytokine is interleukin-15 (IL-15). In some embodiments, the cytokine is mutant IL-15. In one such embodiment, IL-15 is IL-15[N27D]. In some embodiments, the cytokine is interleukin-7 (IL-7). In some embodiments, the cytokine is interleukin-9 (IL-9). In some embodiments, the cytokine is interleukin-10 (IL-10). In some embodiments, the cytokine is interleukin-21 (IL-21). In some embodiments, the cytokine is interleukin-10 (IL-10).

[0403] In some embodiments of the (XII) conjugate, D 1 It is an anticancer agent. In some such implementations, D 1 For SN-38. In some such implementations, D 1 It is monomethylolpropionate E. In some such embodiments, D 1 For capecitabine. In some such implementations, D 1 For irinotecan. In some such implementations, D 1 It is trofluuridine. In some such embodiments, D 1 For Doxorubic stars. In some such implementations, D 1 For Dorcetathione. In some such implementations, D 1 It is Infiniti.

[0404] In some embodiments of the (XII) conjugate, D 1 It is a peptide.

[0405] In some embodiments, the peptide is RGK-MCA. In some embodiments, the peptide is neurotensin. In some embodiments, the peptide is gonadotropin-releasing hormone (GnRH). In some embodiments, the peptide is GIP (1-42) human peptide. In some embodiments, the peptide is D6-GnRH. In some embodiments, the peptide is insulin. In some embodiments, the peptide is teriparatide. In some embodiments, the peptide is salmon calcitonin. In some embodiments, the peptide is iparene. In some embodiments, the peptide is vasopressin. In some embodiments, the peptide is glucagon. In some embodiments, the peptide is alexiside. In some embodiments, the peptide is goserelin. In some embodiments, the peptide is leuprolide. In some embodiments, the peptide is teriparatide. In some embodiments, the peptide is lixinaside. In some embodiments, the peptide is desmopressin. In some embodiments, the peptide is oxytocin. In some embodiments, the peptide is corticotropin. In some embodiments, the peptide is sermorelin. In some embodiments, the peptide is bumenotide. In some embodiments, the peptide is abalotropic peptide. In some embodiments, the peptide is dilorelin.

[0406] In some embodiments, the peptide drug is a glucagon-like peptide-1 (GLP-1) receptor agonist used to treat obesity and / or diabetes. In some such embodiments, the peptide drug is semaglutide, liraglutide, dulaglutide, abiglutide, or exenatide. Because the duration of action of the peptide drug is significantly prolonged when incorporated into the conjugates of this disclosure, GLP-1 agonist conjugates can reduce the frequency of administration, such as once weekly, once every two weeks, once every three weeks, or once monthly.

[0407] In some embodiments, the peptide drug is a glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic peptide (GIP) receptor agonist used to treat obesity and / or diabetes. In some such embodiments, the peptide drug is telposide. Because the duration of action of the peptide drug is significantly prolonged when incorporated into the conjugates of this disclosure, the GLP-1 / GIP agonist conjugates can reduce the frequency of administration, such as once weekly, once every two weeks, once every three weeks, or once monthly.

[0408] In some embodiments, the peptide is an endogenous opioid peptide. In some such embodiments, the peptide is endorphin-1 or endorphin-2. In other embodiments, the peptide is corticomerol.

[0409] In some embodiments of the (XII) conjugate, D 1 It is an opioid antagonist. In some embodiments, the opioid antagonist is naloxone, nalmefen, natrozone, methylnaltrozone, nalbuphine, or nalprofen.

[0410] In some embodiments of the (XII) conjugate, D 1 It is an opioid agonist. In some embodiments, the opioid agonist is morphine, oxycodone, hydromorphone, or methadone.

[0411] In some embodiments, the conjugate is selected from the conjugates in Table A or their salts.

[0412] Table A.

[0413] Pharmaceutical Composition This document provides pharmaceutical compositions comprising one or more conjugates of this disclosure (e.g., conjugates of formulas (I) to (XII) or (II') to (VIII') or any variations or embodiments thereof described elsewhere herein) or a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, this document provides a pharmaceutical composition comprising (i) a compound of any of formulas (I) to (XII) or (II') to (VIII') or any variations or embodiments thereof described elsewhere herein, or a pharmaceutically acceptable salt thereof, and (ii) one or more pharmaceutically acceptable excipients.

[0414] Suitable pharmaceutically acceptable excipients may include, for example, fillers, diluents, sterile aqueous solutions and various organic solvents, permeation enhancers, solubilizers and adjuvants. The term excipient may cover a wide range of substances, including but not limited to any substance used as a binder, disintegrant, coating, compression / encapsulation aid, cream or lotion, lubricant, solution for parenteral administration, material for chewable tablets, sweetener or flavoring agent, suspending / gelling agent or wet granulation agent. Examples of suitable excipients are well known to those skilled in the art. See, for example... Handbook of Pharmaceutical Excipients ,Pharmaceutical Press (2017), which is incorporated herein by reference in its entirety. Such compositions are prepared in a manner well known in pharmaceutical technology. See, for example Remington's Pharmaceutical Sciences , Academic Press, 23rd edition (2020), which is incorporated in its entirety by reference.

[0415] Treatment In one aspect, this document provides a method for increasing the plasma circulation time of an active agent, the method comprising covalently linking the active agent to a plasma protein conjugate to form a conjugate of the present disclosure (e.g., conjugates of formulas (I) to (XII) or (II') to (VIII') or any variation thereof).

[0416] In one aspect, this document provides a method for increasing the in vivo half-life of an active agent, the method comprising covalently linking the active agent to a plasma protein binder to form a conjugate of the present disclosure (e.g., conjugates of formulas (I) to (XII) or (II') to (VIII') or any variation thereof).

[0417] In one aspect, this document provides a method for increasing the duration of action (DOA) of an active agent, the method comprising covalently linking the active agent to a plasma protein binder to form a conjugate of this disclosure.

[0418] In one aspect, this document provides a method for reducing the immunogenicity of an active agent in a subject, the method comprising obtaining a delivery system having a ligand and a linker, said ligand (i) having high selectivity for an endogenous plasma protein of the subject, the plasma protein having a molecular weight ranging from about 30 kDa to about 80 kDa; (ii) having high binding affinity for the plasma protein, with a Kd of at least 10⁻⁶ M; and (iii) having a molecular weight ranging from about 200 Da to about 2000 Da; said linker having a length ranging from about 10 angstroms to about 50 angstroms or 10 atoms to 50 atoms. The method further comprises covalently linking the delivery system to the active agent to produce a conjugated active agent; and administering said conjugated active agent to the subject; wherein the plasma protein protects the active agent from antibody production in the subject after administration.

[0419] In one aspect, this document provides methods for treating diseases, symptoms, or ailments, the methods comprising administering to an individual in need a conjugate of the present disclosure (e.g., a conjugate of any of formulas (I) to (XII) or (II') to (VIII') or any variation or embodiment thereof, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt thereof), or a pharmaceutical composition comprising a conjugate of the present disclosure, any variation or embodiment thereof, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients.

[0420] In some implementation methods, a therapeutically effective amount of the compound is administered.

[0421] In some embodiments, when administered to a subject using the delivery system provided herein, the half-life of the active agent (opioid antagonist or opioid agonist) can range from about 30 minutes to about 90 days. In some embodiments, when administered to a subject using the delivery system provided herein, the half-life of the active agent (opioid antagonist or opioid agonist) can range from about 30 minutes to about 30 days. For example, in some embodiments, when administered to a subject using a delivery system of the ligand with high transthyretin selectivity as taught herein, the half-life of the active agent can range from about 30 minutes to about 2 days. Similarly, in some embodiments, when administered to a subject using a delivery system of the ligand with high human serum albumin selectivity as taught herein, the half-life of the active agent can range from about 30 minutes to about 2 days. Therefore, in some implementations, when the delivery system taught herein is administered to a subject, the half-life of the active agent may be about 30 minutes, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 6 hours, about 8 hours, about 10 hours, about 12 hours, about 18 hours, about 24 hours, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 10 days, about 12 days, about 16 days, about 20 days, about 30 days, or any amount in increments of 1 hour. Those skilled in the art will understand that the half-life of the activity of an active agent administered to a subject using the delivery system taught herein may be within the following ranges: about 30 minutes to about 2 days, about 1 hour to about 36 hours, about 2 hours to about 20 hours, about 2 hours to about 10 hours, about 3 hours to about 30 hours, about 3 hours to about 15 hours, about 4 hours to about 40 hours, about 4 hours to about 20 hours, about 5 hours to about 50 hours, about 5 hours to about 25 hours, or any range or amount thereof in increments of 1 hour.

[0422] In one aspect, this article provides a method for reducing the risk of recurrent respiratory depression, the method comprising administering the conjugate of the present disclosure to an individual in need prior to exposure to a drug.

[0423] In one aspect, this document provides a preventative method comprising administering the disclosed conjugate to an individual in need prior to exposure to the drug.

[0424] In some embodiments, the conjugates of this disclosure are administered parenterally to a subject in need. For example, the conjugates of this disclosure may be administered subcutaneously, intravenously, intramuscularly, or intrathecally.

[0425] Partly due to the hydrophilicity of the TTR selective ligand, the conjugates of this disclosure (especially those of formulas (II) to (XII) or (II') to (VIII') exhibit enhanced tissue (especially subcutaneous) absorption. Following tissue (e.g., subcutaneous) absorption, the reversible binding of the TTR selective ligand to the conjugate significantly prolongs the duration of action of the active agent. As illustrated herein, cleavage of the linker between the TTR selective ligand and the active agent (opioid antagonist or opioid agonist) releases the active agent, thus allowing it to exert its pharmacological effects.

[0426] Furthermore, it has been found that the release of the active agent can be effectively controlled by appropriately selecting the cleavable moieties in the conjugate. For example, certain ester moieties described herein (e.g., PABA or PHBA) undergo slow hydrolysis upon administration, thereby releasing the active agent in a controlled manner for a prolonged period. Slower hydrolysis and a high binding TTR effectively prolong the plasma circulation time of the active agent.

[0427] Therefore, the conjugates of this disclosure exhibit several beneficial properties of novel prodrug methods. The balanced hydrophilicity of the conjugates allows for rapid absorption from tissues into the bloodstream, the binding of TTR prolongs the half-life, and the cleavable moiety allows for modulated drug release. The desired release rate of a specific active drug from the conjugates of this disclosure will vary depending on the active agent.

[0428] In some embodiments, the DOA of the drug is at least 12 hours after administration of the conjugates of this disclosure. In some embodiments, the DOA of the drug is at least 20 hours after administration of the conjugates of this disclosure. In some embodiments, the DOA of the drug is at least 24 hours after administration of the conjugates of this disclosure. In some embodiments, the DOA of the drug is at least 30 hours after administration of the conjugates of this disclosure. In some such embodiments, the DOA of the drug is at least 40 hours after administration of the conjugates of this disclosure. In some such embodiments, the DOA of the drug is at least 48 hours after administration of the conjugates of this disclosure. In some such embodiments, the DOA of the drug is at least 72 hours after administration of the conjugates of this disclosure. In some such embodiments, the DOA of the drug is at least 96 hours after administration of the conjugates of this disclosure. In some such embodiments, the DOA of the drug is at least 1 week after administration of the conjugates of this disclosure. In some such embodiments, the DOA of the drug is at least 2 weeks after administration of the conjugates of this disclosure. In some such embodiments, the DOA of the drug is at least 3 weeks after administration of the conjugate of this disclosure. In some such embodiments, the DOA of the drug is at least 1 month after administration of the conjugate of this disclosure. In some such embodiments, the DOA of the drug is from about 12 hours to about 48 hours after administration of the conjugate of this disclosure. In some such embodiments, the DOA of the drug is from about 24 hours to about 48 hours after administration of the conjugate of this disclosure. In some embodiments, the DOA of the drug is from about 48 hours to about 96 hours after administration of the conjugate of this disclosure. In some embodiments, the DOA of the drug is from about 24 hours to about 1 week after administration of the conjugate of this disclosure.

[0429] In some embodiments, the plasma circulation of the drug is at least 12 hours after administration of the conjugate of this disclosure. In some embodiments, the plasma circulation of the drug is at least 20 hours after administration of the conjugate of this disclosure. In some embodiments, the plasma circulation of the drug is at least 24 hours after administration of the conjugate of this disclosure. In some embodiments, the plasma circulation of the drug is at least 30 hours after administration of the conjugate of this disclosure. In some such embodiments, the plasma circulation of the drug is at least 40 hours after administration of the conjugate of this disclosure. In some such embodiments, the plasma circulation of the drug is at least 48 hours after administration of the conjugate of this disclosure. In some such embodiments, the plasma circulation of the drug is at least 72 hours after administration of the conjugate of this disclosure. In some such embodiments, the plasma circulation of the drug is at least 96 hours after administration of the conjugate of this disclosure. In some such embodiments, the plasma circulation of the drug is at least 1 week after administration of the conjugate of this disclosure. In some such embodiments, the plasma circulation of the drug is at least 2 weeks after administration of the conjugate of this disclosure. In some such embodiments, the plasma circulation of the drug is at least 3 weeks after administration of the conjugate of this disclosure. In some such embodiments, the plasma circulation of the drug is at least 1 month after administration of the conjugate of this disclosure. In some such embodiments, the plasma circulation of the drug is from about 12 hours to about 48 hours after administration of the conjugate of this disclosure. In some such embodiments, the plasma circulation of the drug is from about 24 hours to about 48 hours after administration of the conjugate of this disclosure. In some embodiments, the plasma circulation of the drug is from about 48 hours to about 96 hours after administration of the conjugate of this disclosure. In some embodiments, the plasma circulation of the drug is from about 24 hours to about 1 week after administration of the conjugate of this disclosure.

[0430] In some embodiments, the active pharmaceutical ingredient (D) delivered via the conjugate of this disclosure 1 The active pharmaceutical ingredient (API) is a small molecule, nucleotide, oligonucleotide, polynucleotide, amino acid, oligopeptide, polypeptide, or protein. It may include, but is not limited to, antiproliferative agents, antitumor agents, antimitotic agents, anti-inflammatory agents, antiplatelet agents, anticoagulants, antifibrinogens, antithrombins, antibiotics, antiallergens, and antioxidants.

[0431] Antiproliferative agents include, for example, actinomycin D, actinomycin IV, actinomycin I1, actinomycin X1, actinomycin C1, and dermatomycin (Cosmegen.RTM., Merck & Co., Inc.). Anti-tumor or anti-mitotic agents include, for example, paclitaxel (TAXOL, Bristol-Myers Squibb Co.), docetaxel (TAXOTERE, Aventis SA), methotrexate, irinotecan, SN-38, azathioprine, vincristine, vinblastine, fluorouracil, doxorubicin hydrochloride (ADRIAMYCIN, Pfizer, Inc.), and mitomycin (MUTAMYCIN, Bristol-Myers Squibb Co.) and any of their prodrugs, codrugs, metabolites, analogues, homologs, derivatives, salts, and combinations. Antiplatelet agents, anticoagulants, antifibrinogens, and antithrombins include, for example, sodium heparin, low molecular weight heparin, heparin-like substances, hirudin, argatroban, forskolin, vapiprost, prostacyclin and prostacyclin analogs, polydextrose, D-phe-pro-arg-chloromethyl ketone (synthetic antithrombin), dipyridamole, glycoprotein IIb / IIIa platelet membrane receptor antagonist antibodies, recombinant hirudin, and thrombin inhibitors (ANGIOMAX, Biogen, Inc.) and any prodrugs, combinations thereof, metabolites, analogs, homologs, derivatives, salts, and combinations thereof.Cell growth inhibitors or antiproliferative agents include, for example, angiopeptin; angiotensin-converting enzyme inhibitors, such as captopril (CAPOTEN and CAPOZIDE, Bristol-Myers Squibb Co.), cilazapril, or lisinopril (PRINVIL ​​and PRINZIDE, Merck & Co., Inc.); calcium channel blockers, such as nifedipine; colchicine; fibroblast growth factor (FGF) antagonists, fish oil (ω3-fatty acids); histamine antagonists; and lovastatin (MEVACOR, Merck & Co., Inc.). Inc.; monoclonal antibodies, including but not limited to antibodies specific to platelet-derived growth factor (PDGF) receptors; nitroprusside; phosphodiesterase inhibitors; prostaglandin inhibitors; suramin; serotonin blockers; steroids; thioprotease inhibitors; PDGF antagonists, including but not limited to triazolopyrimidine; and nitric oxide, and any of their prodrugs, combinations, metabolites, analogs, homologs, homologous drugs, derivatives, salts, and combinations. Antihistamines include, but are not limited to, pemirolast potassium (ALAMAST, Santen, Inc.) and any of its prodrugs, combinations, metabolites, analogs, homologs, homologous drugs, derivatives, salts, and combinations.

[0432] In some embodiments, the conjugates of this disclosure may be administered in combination with a second drug. For example, in some embodiments, the conjugates of this disclosure may be administered with a second drug (D) in its unconjugated form. 2 ) Combined administration. The second drug (D) in its unconjugated form. 2 Drugs that can be conjugated (D) 1 (Same or different.) In some implementations, D 1 and D 2 Both are anti-cancer drugs.

[0433] In some embodiments, the conjugates of this disclosure are administered subcutaneously to human subjects at a dose of about 0.005 mg / kg to about 40 mg / kg. In some embodiments, the conjugates of this disclosure are administered subcutaneously to human subjects at a dose of about 0.01 mg / kg to about 40 mg / kg. In some embodiments, the conjugates of this disclosure are administered subcutaneously to human subjects at a dose of about 0.005 mg / kg to about 10 mg / kg. In some embodiments, the conjugates of this disclosure are administered subcutaneously to human subjects at a dose of about 0.1 mg / kg to about 10 mg / kg. In some embodiments, the conjugates of this disclosure are administered subcutaneously to human subjects at a dose of about 0.5 mg / kg to about 10 mg / kg. In some embodiments, the conjugates of this disclosure are administered subcutaneously to human subjects at a dose of about 5 mg / kg to about 40 mg / kg. In some embodiments, the conjugates of this disclosure are administered subcutaneously to human subjects at a dose of about 10 mg / kg to about 30 mg / kg. In some embodiments, the conjugates disclosed herein are administered subcutaneously to human subjects at a dose of about 15 mg / kg to about 25 mg / kg.

[0434] medicine box This disclosure further provides a pharmaceutical kit for implementing the methods disclosed herein. The kit may contain the conjugates of this disclosure described herein or pharmaceutically acceptable salts thereof, along with suitable packaging. The kit may contain one or more containers containing any of the compounds described herein. In one aspect, the kit includes a compound of this disclosure or a pharmaceutically acceptable salt thereof, and a label and / or instructions for using said compound to treat the disease or condition described herein. The kit may contain a unit dosage form of the compound.

[0435] This document provides a kit comprising (i) a conjugate of the present disclosure, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a compound of formula (I) or any variation or embodiment thereof, or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients, and (ii) instructions for therapeutic use.

[0436] In some implementation schemes, the individual is defined as a human being.

[0437] Articles are also provided, wherein the articles comprise, in a suitable container, a conjugate thereof, any variation or embodiment thereof, a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt thereof.

[0438] In some implementations, the container may be a vial, a wide-mouth bottle, an ampoule, a pre-loaded syringe, or an intravenous bag.

[0439] Preparation method This document provides methods for preparing the conjugates of this disclosure, or any variations or embodiments thereof, or pharmaceutically acceptable salts of any of the foregoing. The synthetic methods described below, as well as those detailed in the examples, are merely illustrative of some methods by which the compounds of this disclosure, or embodiments or aspects thereof, can be synthesized. As will be apparent to those skilled in the art, various modifications can be made to these synthetic reaction schemes. Although certain exemplary embodiments are depicted and described herein, the compounds of this disclosure, or any variations or embodiments thereof, can be prepared using suitable starting materials according to the methods generally described herein and / or methods available to those skilled in the art.

[0440] When a specific stereoisomer or non-specific stereoisomer or mixture of stereoisomers is shown in the following general procedure, it should be understood that similar chemical transformations can be performed on other specific stereoisomers or non-specific stereoisomers or mixtures thereof.

[0441] Representative examples of the preparation of the conjugates of this disclosure are provided in Example 1. In some embodiments, the hydroxyl groups of the pharmaceutical moiety may undergo esterification using methods known in the art. In some embodiments, the reactive hydroxyl group is an alkyl hydroxyl group on a small molecule, peptide, or protein. In some embodiments, the reactive hydroxyl group is a phenolic hydroxyl group on a small molecule, peptide, or protein. In some embodiments, a tyrosine residue of the peptide or protein serves as the esterification site. In some embodiments, a serine residue of the peptide or protein serves as the esterification site.

[0442] Where a particular reagent or solvent is specified for a reaction in a general procedure, those skilled in the art will recognize that other reagents or solvents may be substituted as needed.

[0443] Example The synthetic reaction schemes detailed in the examples are merely illustrative of some methods by which the disclosed compounds or embodiments or aspects thereof may be synthesized. As will be apparent to those skilled in the art, various modifications can be made to these synthetic reaction schemes.

[0444] The starting materials and intermediates of the synthetic reaction scheme can be separated and purified using conventional techniques, including but not limited to filtration, distillation, crystallization, and chromatography, when necessary. These materials can be characterized using conventional methods that include physical constants and spectral data.

[0445] Although certain exemplary embodiments are depicted and described herein, the compounds of this disclosure or any variations or embodiments thereof may be prepared using appropriate starting materials in accordance with the methods generally described herein and / or methods available to those skilled in the art.

[0446] All publications, patent applications, patents and other references mentioned in this article are explicitly incorporated as a whole by reference, as if each reference were incorporated individually by reference.

[0447] Example 1: Synthesis and Characterization of the Conjugates of this Disclosure Materials and methods Materials and reagents. Naloxone hydrochloride (#N285000) was purchased from Toronto Research Chemicals, Toronto, Ontario. Nalmefen hydrochloride (#408550) was purchased from Tocris Bioscience™. Naloxone-D5 100µg / mL (#N-063) was purchased from Cerilliant. Morphine sulfate (CII) (#M1167) and fentanyl citrate (CII) (#F1147) were purchased from Spectrum Chemical Manufacturing Corporation. Digoxin saponins (#D0540) and loperamide hydrochloride (#L0154) were purchased from Fisher Scientific. Dodecyl azide (#A1887) was purchased from Synthonix. Prealbumin from human plasma (human TTR) (#P1742) and prealbumin from human serum (#H4522) were purchased from Sigma Aldrich. Innovative-grade US-derived rat plasma (#IGRTSDPLANAH), rabbit plasma (#IGRBPLANAH), cynomolgus monkey plasma (#IGMNCYPLANAH), and mixed human plasma (#IPLAWBNAH) were purchased from Innovative Research. XTreme 200 mixed-sex human liver microsomes (#1910096), male IGS SD rat liver microsomes (#2210274), male cynomolgus monkey liver microsomes (#2210248), and the NADPHRapidStart regeneration system for prolonging metabolism (#K5100-5) were purchased from Xenotech. Human carboxylesterase 1 (CES1, #E0287-1VL) and carboxylesterase 2 (CES2, #C4749) were purchased from Sigma Aldrich. RPMI-1640 medium and minimum Eagle's medium (HyClone, Utah, USA), fetal bovine serum (Gemini), penicillin / streptomycin (100 units / mL and 100 µg / mL, Gibco, NY, USA), and L-glutamine (Glutamax-100X, #35-050-061, Gibco, NY, USA) were purchased from the companies mentioned in parentheses. CellBIND® 96-well clear plates (#CLS3340-50EA) were purchased from Corning®.The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazol bromide) of the test compound in the cell line was determined using the CellTiter 96 non-radioactive cell proliferation assay kit (#G4000, Promega, WI, USA).

[0448] Chemical synthesis and HPLC purity analysis General chemical procedure. Unless otherwise specified, all reactions were carried out under an argon or nitrogen atmosphere, using anhydrous solvents under anhydrous conditions. Solvents used were ACS grade from Fisher. Unless otherwise specified, yields refer to chromatographic and spectroscopic results. 1 H NMR and 13 Homogeneous material (C10 NMR). Reagents were purchased from Aldrich and Fisher and used directly without further purification. UV light was used, and an iodine chamber was used as the imaging agent, via a coating of fluorescent indicator F1000 nm. 254 Thin-layer chromatography (TLC) was performed on an EMD Millipore® silica gel 60 plastic TLC plate (#1.05735.0001) to monitor the reaction. Normal-phase rapid column chromatography was performed using a Combi Flash® Rf+Lumen instrument (Teledyne ISCO) with a high-performance silica rapid column (RediSep® Rf+ Gold), and preparative thin-layer chromatography (PTLC) separations were performed on an Analtech® 2 mm (60F-254) (#P02015) spectrometer. Recordings were performed on a JEOL JNM-ECA 600 spectrometer. 1 H NMR and 13 C10 NMR spectroscopy was performed, and the residual undeuterated solvent was used as an internal reference for calibration. High-resolution mass spectrometry (HRMS) was performed at the UCBerkley mass spectrometry facility using electrospray ionization from the solution. Coupling constants (J) are expressed in Hertz.

[0449] A preparative HPLC method was used to purify the synthesized compounds. Purification was performed using Waters Chromesope V2.1 software on a Waters 150Q LC system with a PDA and a FlexInject system connected to a photodiode array detector operating in the 190–800 nm UV range. After injecting 5 mL of each sample, HPLC analysis was performed at ambient temperature on a SunFire® Prep C18 OBD column (19 x 250 mm, 5 μm) to obtain a chromatogram at 310 nm UV absorbance. The mobile phase consisted of solvent A and solvent B and was prepared at a flow rate of 20 mL / min. -1The flow rate of the solvent was determined by the following methods: Solvent A consisted of methanol-water (5:95, v / v) containing 0.1% trifluoroacetic acid, and Solvent B consisted of methanol-water (95:5, v / v) containing 0.1% trifluoroacetic acid. The HPLC program was as follows: an isocratic gradient of 0% solvent B for the first 4 min, followed by a linear increase from 0% to 100% solvent B for 4–20 min, then isocratic elution with 100% solvent B until 24 min, followed by a linear decrease to 0% B at 26 min, and then holding at 0% solvent B for 4 min.

[0450] An analytical HPLC method was used to evaluate the purity of synthesized compounds. Detailed HPLC information (trace, retention time, and purity %) for key compounds is included below. Purity (all compounds >95%) of key compounds was analyzed using a C18 reversed-phase HPLC column on an Arc Premier Waters modular HPLC system connected to a Waters 2998 photodiode array detector operating in the 190–800 nm UV range, and quantification was performed using Empower 3 software. After injecting 20 μL of each sample, the purity was measured at 1 mL / min at ambient temperature. -1 Elution was performed by HPLC on a Waters™ XBridge C18 column (4.6 x 250 mm, 5 μm) to obtain chromatograms at 310 nm UV absorbance. The mobile phase consisted of solvent A and solvent B. Solvent A was composed of methanol-water (5:95, v / v) containing 0.1% ammonium hydroxide, and solvent B was composed of methanol-water (95:5, v / v) containing 0.1% ammonium hydroxide. The HPLC program for the C18 column was a gradient method as follows: linear increase from 0% to 100% solvent B over 0–20 min, followed by isocratic elution with 100% solvent B up to 25 min, decreasing to 0% B at 27 min, and then holding at 0% B for 3 min.

[0451] General Procedure A. Preparation of Naloxone Free Base

[0452] At ambient temperature, a tertiary amine base, N,N-diisopropylethylamine (1.25 equivalents), is added to a solution of naloxone hydrochloride (1 equivalent) in dichloromethane. After stirring for a short time (e.g., 5-10 minutes), the solvent is removed under reduced pressure to obtain the corresponding free base of naloxone.

[0453] General Procedure B. Ester Synthesis Under an inert atmosphere (e.g., N2 or argon), the selected drug (D) is... 1The selected carboxylic acid (1.2 equivalents) and a catalytic amount of acylation catalyst (e.g., 4-dimethylaminopyridine, 0.2 equivalents) dissolved in dichloromethane are added. The reaction mixture is stirred briefly to allow pre-activation, followed by the addition of a coupling agent, such as N,N'-dicyclohexylcarbodiimide (approximately 1 equivalent). The reaction mixture is stirred at ambient temperature for an extended period (e.g., 12–24 hr) to ensure complete conversion. The mixture is then filtered and the solvent is removed under reduced pressure. The crude product is purified using chromatographic techniques (e.g., rapid silica gel column chromatography with an appropriate gradient of ethyl acetate and hexane) to obtain the target ester. The structure and purity of the product are confirmed using analytical methods such as nuclear magnetic resonance spectroscopy (NMR) and mass spectrometry (MS).

[0454] General procedure C. Copper (I)-catalyzed azide-alkyne cycloaddition (CuAAC).

[0455] Click (CuAAC) coupling was performed by coupling naloxone-alkyne / drug-alkyne (1.2 equivalents), azide-connector-AG10 / azide-connector (1 equivalent), CuSO4·5H2O (0.5 equivalents), and sodium ascorbate (1.2 equivalents) in a mixture of tetrahydrofuran / water (2:1). The reaction mixture was stirred overnight at room temperature. Tetrahydrofuran and water were removed under reduced pressure. The residue was purified by silica gel chromatography or dissolved in methanol and purified by preparative reversed-phase HPLC to obtain the desired product.

[0456] General procedure D. Removal of Boc protecting groups The Boc-protected conjugate and precursor were treated with a mixture of dichloromethane and trifluoroacetic acid (3:1). The reaction mixture was stirred for 2 hours at room temperature under an inert atmosphere. Next, ammonium hydroxide (28-36%) dissolved in water was slowly added dropwise with methanol. This solution was then concentrated under reduced pressure and purified by preparative reversed-phase HPLC to obtain the desired product.

[0457] General program for E. Fmoc peptide synthesis First, Rink amide resin was swollen in dichloromethane (DCM) to prepare it for peptide synthesis. Fmoc protecting groups were removed using DMF containing 20% ​​piperidine for 25 minutes, followed by thorough washing with DMF. Coupling was performed by pre-activating the Fmoc-protected amino acids (3 equivalents) in DMF with HBTU (3 equivalents) and DIPEA (6 equivalents) for 3 minutes, then adding the mixture to the resin and stirring for 40 minutes. Coupling and Fmoc deprotection were monitored using the Kaiser test to ensure completion. After synthesis, acid-labile protecting groups (e.g., t-Boc and t-Bu) were cleaved and removed from the resin bulk at room temperature for 2 hours using a mixture of 95% trifluoroacetic acid (TFA), 2.5% water, and 2.5% triisopropylsilane (TIPS). The crude peptide / peptide ester conjugate was then precipitated with cold diethyl ether, collected by centrifugation, washed, and dried. The final product was purified using preparative HPLC.

[0458] Synthesis of AG10-L-E2-naloxone Scheme 1 shows a schematic diagram of the synthesis of AG10-L-E2-naloxone.

[0459] Scheme 1. Synthesis of AG10-L-E2-naloxone.

[0460] Ethyl 4-((tert-butoxycarbonyl)(prop-2-yn-1-yl)amino)benzoate (1). Triethylamine (1.2 g, 1.64 mL, density 0.73 g / mL) was added to a solution of ethyl 4-(prop-2-yn-1-yl)benzoate (2 g, 9.84 mmol, 1 equivalent), di-tert-butyl dicarbonate (2.6 g, 11.9 mmol, 1.2 equivalent), and 4-dimethylaminopyridine (0.24 g, 1.96 mmol, 0.2 equivalent) in anhydrous tetrahydrofuran (12 mL) under a nitrogen atmosphere at room temperature. -1 11.86 mmol, 1.2 equivalents). The reaction mixture was stirred overnight at 80°C. The reactants were cooled to room temperature, and then the mixture containing formic acid (906 mg, 0.742 mL, density 1.22 g / mL) was added at room temperature. -1 3 mL of ethyl acetate (19.68 mmol, 2 equivalents) was added dropwise to a stirred reaction vessel. The solvent and excess formic acid were then dried under reduced pressure. The remaining residue was dissolved in 30 mL of ethyl acetate, filtered, and the solvent was removed under reduced pressure. The residue was purified by rapid column chromatography (silica gel, 0-100% ethyl acetate / hexane) to give compound 1 (2.76 g, 92.4% separation yield) as a transparent yellow viscous liquid. 1H NMR (600 MHz, CDCl3) δ 7.98 (d,2H, J = 9), 7.38 (d, 2H, J = 9), 4.39 (s, 2H), 4.33 (q, 2H, J = 7.2), 2.24 (s, 1H), 1.44 (s, 9H), 1.35 (t, 3H, J = 7.2); 13 C-NMR (150 MHz, CDCl3) δ 166.12, 153.39,146.16, 130.16, 127.75, 125.23, 100.00, 81.83, 79.64, 72.22, 61.02, 39.49,28.29, 14.40. HRMS (DART) m / z: C 17 H 21 NO4[M+Na] + The calculated value is 326.1363; the experimental value is 326.1364.

[0461] (4aS,7aR,12bS)-4-((tert-butoxycarbonyl)(prop-2-yn-1-yl)amino)benzoic acid 3-allyl-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methylbridged benzofurano[3,2-e]isoquinoline-9-yl ester (2). Water (3.75 mL) containing sodium hydroxide (729 mg, 18.2 mmol, 2 equivalents) was added to a stirred solution of 1 (2.76 g, 9.1 mmol, 1 equivalent) in methanol (7.5 mL) at room temperature. This reaction mixture was stirred overnight at 50 °C, and then the water and methanol solvent were dried under reduced pressure. The remaining residue was treated with a solution containing formic acid (2.44 g, 2 mL, density 1.22 g / mL). -1 The acetic acid was redissolved in ethyl acetate (30 mL) at 53 mmol, filtered, and the solvent was removed under reduced pressure. The residue was purified by rapid column chromatography (silica gel, 0-100% ethyl acetate / hexane) to give a white powder of the acid hydrolysis product of 1 (2.46 g), which was used directly in the next step. N,N-diisopropylethylamine (0.88 g, 1.2 mL, density 0.742 g / mL) was added to a solution of naloxone hydrochloride (1.32 g, 3.63 mmol, 1 equivalent) in CH2Cl2 (30 mL) at room temperature. -14.5 g (1.25 mmol, 1.25 equivalents) was added to the mixture and stirred for five minutes. The CH2Cl2 was then removed under reduced pressure, leaving a dry residue. At room temperature under a nitrogen atmosphere, a solution of the acid hydrolysis product of 1 (1.2 g, 4.36 mmol, 1.2 equivalents) and 4-dimethylaminopyridine (89 mg, 0.72 mmol, 0.2 equivalents) dissolved in 30 mL of CH2Cl2 was added to this residue. The reaction mixture was stirred for five minutes, followed by the addition of N-N'-dicyclohexylcarbodiimide (0.75 g, 3.63 mL, 3.63 mmol, 1 equivalent) to a 1 M solution of anhydrous CH2Cl2 under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 48 hr, then filtered and excess CH2Cl2 was removed under reduced pressure. The residue was purified by rapid column chromatography (silica gel, 0-100% ethyl acetate / hexane) to give compound 2 (1.5 g, 70% separation yield) as a white powder. 1 H NMR (600MHz, CDCl3) δ 8.16 (d, 2H, J = 7.8), 7.44 (d, 2H, J = 8.4), 6.93 (d, 1H, J = 8.4), 6.71 (d, 1H, J = 8.4), 5.83-5.75 (m, 1H), 5.28-5.15 (m, 2H), 4.68 (s,1H), 4.41(s,2H), 3.20-3.12 (m, 3H), 3.04-2.96 (m, 2H), 2.66-2.57 (m, 2H), 2.43-2.36(m, 1H), 2.31-2.24 (m, 2H), 2.19-2.11 (m, 1H), 1.88-1.82 (m, 1H), 1.67-1.59(m, 2H), 1.46 (s, 9H); 13 C-NMR (150 MHz, CDCl3) δ 207.82, 163.73, 153.47,148.11, 146.93, 135.19, 132.86, 131.27, 130.52, 130.34, 126.30, 125.43,123.20, 119.63, 118.44, 100.00, 90.78, 82.08, 79.62, 72.59, 70.24, 62.06,57.70, 50.80, 43.38, 39.51, 36.25, 31.34, 30.74, 28.42, 23.13; HRMS (DART)m / z： C 34 H 36 N₂O₇[M+H] + The calculated value is 585.2595; the experimental value is 585.2593.

[0462] 3-((6-((2-(2-(2-(4-(((4-((((4aS,7aR,12bS)-3-allyl-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methylbridged benzofurano[3,2-e]isoquinoline-9-yl)oxy)carbonyl)phenyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl)amino)hexyl)oxy)-5-(3-(3,5-dimethyl-1H-pyrazol-4-yl)propoxy)benzoic acid (AG10-L-E2-naloxone). Click (CuAAC) coupling was performed by coupling compound 2 (561 mg, 0.96 mmol, 1.2 equivalents), compound 3 (515 mg, 0.8 mmol, 1 equivalent; synthesized as previously reported), CuSO4·5H2O (100 mg, 0.4 mmol, 0.5 equivalents), and sodium ascorbate (190 mg, 0.96 mmol, 1.2 equivalents) in a) 9 mL of a mixture of tetrahydrofuran / water (2:1). The reaction mixture was stirred overnight at room temperature. Tetrahydrofuran and water were removed under reduced pressure. The residue was dissolved in water and extracted with CH2Cl2. The organic extract was concentrated under reduced pressure and purified by rapid column chromatography (silica gel, 0-100% CH2Cl2 / methanol) to give a di-Boc protected AG10-L-E2-naloxone intermediate, which was directly treated with a mixture of 4 mL CH2Cl2 / trifluoroacetic acid (3:1). The reaction mixture was stirred for 2 hours at room temperature under an inert atmosphere. Next, ammonium hydroxide (28-36%) dissolved in water was slowly added dropwise along with 3 mL of methanol until no hydrogen formation was observed. This solution was then concentrated under reduced pressure and purified by preparative reversed-phase HPLC to obtain AG10-L-E2-naloxone as a viscous solid (633 mg, 64% separation yield). 1H NMR (600 MHz, CD3OD) δ 7.84-7.79 (m, 3H),7.03-6.99 (m, 2H), 6.95-6.87 (m, 1H), 6.84-6.76 (m, 1H), 6.65-6.60 (m, 2H),6.59-6.54 (m, 1H), 5.90-5.77 (m, 1H), 5.64-5.49 (m, 2H), 4.80 (s, 1H), 4.48-4.42 (m, 1H), 4.40-4.37 (m, 1H), 3.94-3.80 (m, 6H), 3.79-3.72 (m, 2H), 3.56-3.34 (m, 6H), 3.23 (s, 1H), 3.1-2.96 (m, 4H), 2.92-2.82 (m, 3H), 2.75-2.59(m, 2H), 2.59-2.5 (m, 3H), 2.16 (s, 6H), 1.94-1.27 (m, 16H); 13 C-NMR (150 MHz, CD3OD) δ 207.09, 167.44, 163.90, 160.28, 160.22, 153.67, 148.33, 145.07, 133.37, 133.27, 132.33, 128.84, 128.72, 128.17, 125.69, 124.44, 123.78,120.62, 114.76, 111.96, 107.85, 107.80, 106.17, 89.73, 89.69, 70.17, 70.07,69.88, 69.25, 68.15, 67.12, 66.13, 61.90, 55.64, 52.98, 49.85, 48.99, 47.38,46.69, 46.10, 38.21, 35.16, 30.86, 29.64, 28.85, 27.72, 26.15, 25.76, 25.49,23.38, 18.86, 10.62; HRMS (DART) m / z: C 56 H 70 N8O 11 [M+H] + The calculated value is 1031.5237; the experimental value is 1031.5241.

[0463] Synthesis of AG10-L-E2-COOH Scheme 2 shows a schematic diagram of the synthesis of AG10-L-E2-COOH. Scheme 2. Synthesis of AG10-L-E2-COOH.

[0464] 3-((6-((2-(2-(2-(4-((((4-carboxyphenyl)amino)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl)amino)hexyl)oxy)-5-(3-(3,5-dimethyl-1H-pyrazole-4-yl)propoxy)benzoic acid (AG10-L-E2-COOH). To a solution of AG10-L-E2-naloxone (1000 mg, 0.97 mmol, 1 equivalent) in methanol (4 mL), 1 mL of water containing sodium hydroxide (77.6 mg, 1.94 mmol, 2 equivalents) was added. The solution was stirred overnight at 50 °C. The solvent was then removed under reduced pressure. The remaining residue was purified by reversed-phase HPLC to give AG10-L-E2-COOH (420 mg, 60% separation yield); HRMS (DART). m / z: C 37 H 51 N7O8[M+H] + The calculated value is 722.3872; the experimental value is 722.3869.

[0465] Synthesis of C12-L-E2-naloxone Scheme 3 shows a schematic diagram of the synthesis of AG10-L-E2-COOH. Scheme 3. Synthesis of C12-E2-naloxone.

[0466] (4aS,7aR,12bS)-4-(((1-dodecyl-1H-1,2,3-triazol-4-yl)methyl)amino)benzoic acid 3-allyl-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methylbridged benzofurano[3,2-e]isoquinoline-9-yl ester (C12-E2-naloxone). Click (CuAAC) coupling was performed by reacting compound 2 (500 mg, 0.86 mmol, 1 equivalent), dodecyl azide (182 mg, 0.85 mmol, 1 equivalent), CuSO4·5H2O (106.7 mg, 0.425 mmol, 0.5 equivalent), and sodium ascorbate (169.4 mg, 0.855 mmol, 1 equivalent) in a mixture of 3 mL THF / H2O (4:1). The reaction mixture was stirred overnight at room temperature. The solution was concentrated under reduced pressure. The residue was extracted with ethyl acetate. The organic extract was concentrated under reduced pressure to give an intermediate, which was used directly in the next step. Then, a mixture (1 mL) containing TFA and CH2Cl2 (1:4 ratio) was added to the solution of the intermediate, and the reaction mixture was stirred at room temperature for 2 hr. The solution was concentrated under reduced pressure and purified by preparative HPLC to give compound C12-E2-naloxone (255 mg, 43% yield after two steps). 1 HNMR (600 MHz, CD3OD) δ 7.87-7.78 (m, 3H), 6.94 (t, 1H, J = 8.4), 6.85-6.78(m, 1H), 6.61 (s, 2H), 5.91-5.79 (m, 1H), 5.64-5.52 (m, 2H), 4.82 (s, 1H), 4.58-4.34 (m, 1H), 4.27 (t, 2H, J = 7.2), 3.92-3.66 (m, 2H), 3.57-3.32 (m, 2H), 3.17-2.50 (m, 3H), 2.21-1.46 (m, 9H), 1.26-1.06 (m, 18H), 0.79 (t, 3H, J= 7.2); 13C-NMR (150 MHz, CD3OD) δ 206.56, 164.97, 148.36, 133.63, 133.04,131.95, 128.21, 127.65, 126.50, 125.52, 125.30, 124.27, 123.05, 120.21,119.64, 111.58, 97.09, 93.49, 89.43, 69.73, 62.55, 55.47, 50.14, 49.02,46.31, 33.42, 31.73, 29.89, 29.39, 29.19, 28.69, 27.59, 26.97, 26.08, 25.41,24.73, 22.93, 22.40, 13.12; HRMS (DART) m / z: C 41 H 53 N5O5[M+H] + The calculated value is 696.4119; the experimental value is 696.4121.

[0467] Synthesis of AG10-L-E1-naloxone Scheme 4 shows a schematic diagram of the synthesis of AG10-L-E1-naloxone. Scheme 4. Synthesis of AG10-L-E1-naloxone.

[0468] 3-((6-((2-(2-(2-(4-(3-(((4aS,7aR,12bS)-3-allyl-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methylbridged benzofurano[3,2-e]isoquinoline-9-yl)oxy)-3-oxopropyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl)amino)hexyl)oxy)-5-(3-(3,5-dimethyl-1H-pyrazol-4-yl)propoxy)benzoic acid (AG10-L-E1-naloxone). N,N-diisopropylethylamine (0.24 mL, 1.374 mmol, 1 equivalent) was added to a solution of naloxone hydrochloride (500 mg, 1.374 mmol, 1 equivalent) in CH₂Cl₂ (4 mL) at room temperature and stirred for 10 min. The solution was concentrated under reduced pressure to obtain naloxone, which was used directly in the next step. 4-pentyneic acid (134.8 mg, 1.374 mmol, 1 equivalent) and DMAP (33.6 mg, 0.275 mmol, 0.2 equivalent) were added to CH₂Cl₂ (3 mL) containing naloxone under an argon atmosphere. Then, DCC (0.46 mL, 2.748 mmol, 2 equivalent) was added to the solution under an argon atmosphere and the reaction mixture was stirred overnight at room temperature. The reaction mixture was purified by rapid column chromatography (silica gel, 0-100% ethyl acetate / hexane) and the intermediate was used directly in the next step. Click (CuAAC) coupling was performed by coupling the residue with deBoc compound 3 (300 mg, 0.549 mmol, 1 equivalent; previously reported), CuSO4·5H2O (68.5 mg, 0.2745 mmol, 0.5 equivalent), and sodium ascorbate (108.8 mg, 0.549 mmol, 1 equivalent) in a mixture of 3 mL THF / H2O (4:1). The reaction mixture was stirred overnight at room temperature. The solution was concentrated under reduced pressure and purified by preparative HPLC to give compound AG10-L-E1-naloxone (250 mg, 47% yield after two steps). 1H NMR (600 MHz, DMSO-d6)δ 7.95 (s, 1H), 7.05-6.95 (m, 3H), 6.84 (d, 1H, J = 8.4), 6.72-6.68 (m, 1H),5.95-5.87 (m, 1H), 5.62-5.54 (m, 2H), 5.11 (s, 1H), 4.52-4.46 (m, 2H), 4.03-3.91 (m, 5H), 3.86-3.76 (m, 3H), 3.65 (d, 1H, J = 6), 3.64-3.61 (m, 2H), 3.59-3.49 (m, 5H), 3.24-3.16 (m, 1H), 3.12-3.04 (m, 3H), 3.04-2.80 (m, 7H), 2.70-2.56 (m, 1H), 2.5 (s, 1H), 2.29-2.08 (m, 8H), 2.06-1.91 (m, 1H), 1.87-1.80 (m, 2H), 1.75-1.66 (m, 2H), 1.65-1.57 (m, 2H), 1.55-1.15 (m, 7H); 13 C-NMR(150 MHz, DMSO-d6) δ 206.93, 170.46, 167.45, 160.28, 158.82, 158.60, 147.92,145.46, 133.38, 132.69, 129.15, 128.96, 128.14, 125.67, 124.18, 123.07,120.70, 107.81, 106.19, 89.91, 70.13, 70.07, 69.91, 69.28, 68.15, 67.10,66.14, 61.85, 55.63, 55.44, 49.81, 49.03, 47.38, 46.65, 46.06, 35.16, 33.41,30.88, 29.49, 28.86, 27.79, 26.15, 25.81, 25.49, 23.38, 21.18, 18.73, 10.42. HRMS (DART) m / z: C 51 H 67 N7O 11 [M+H] + The calculated value is 954.4971; the experimental value is also 954.4971.

[0469] Synthesis of AG10-short-L-E2-naloxone According to general procedure B and Then, according to the general program C, use Then, AG10-short-L-E2-naloxone was prepared according to the general procedure D.

[0470] Synthesis of AG10-L-PHBA-E-naloxone According to general procedure B and Then, according to the general program C, use Then, AG10-L-PHBA-E-naloxone was prepared according to the general procedure D.

[0471] Synthesis of AG10-L-m-amino-L-naloxone According to general procedure B and Then, according to the general program C, use Preparation of AG10-L-m-amino-E-naloxone.

[0472] Synthesis of PEG-connector-E2-naloxone According to general procedure B and Then, according to the general program C, use Then, PEG connector-E2-naloxone was prepared according to general procedure D.

[0473] Synthesis of AG10-L-E2-propofol According to general procedure B and Then, according to the general program C, use Then, AG10-L-E2-propofol was prepared according to the general procedure D.

[0474] Synthesis of AG10-L-E2-acetaminophen According to general procedure B and Then, according to the general program C, use Then, AG10-L-E2-acetaminophen was prepared according to the general procedure D.

[0475] Synthesis of AG10-L-E2-estrone According to general procedure B and Then, according to the general program C, use Then, AG10-L-E2-estrone was prepared according to the general procedure D.

[0476] Synthesis of AG10-L-E2-Tyr-Ala According to general procedure B and Then, according to general procedure D, then according to general procedure C, use... Prepare AG10-L-E2-Tyr-Ala.

[0477] AG10-L-E2 Endorphin-2 Synthesis According to general procedure B and Then, according to general procedure D, then according to general procedure C, use... Preparation of endorphin-2. Endorphin-2 and di-tert-butyl dicarbonate were prepared at approximately 80°C in the presence of THF containing triethylamine and DMAP. .

[0478] Synthesis of AG10-L-E2-Pepto-Peptide AG10-L-E2-picorphin was prepared according to general procedure E. Scheme 5 illustrates the synthesis of AG10-L-E2-picorphin.

[0479] Scheme 5. Synthesis of AG10-L-E2-Pepto-Peptide Analytical HPLC method. The test compounds were analyzed using a Waters™ XBridge C18 column (4.6 x 250 mm, 5 μm) on an Arc Premier Waters modular HPLC system connected to a Waters 2998 photodiode array detector operating in the 190–800 nm UV range. HPLC analysis was performed using a gradient method with linear increases in 0–100% solvent B over 25 min. The mobile phase consisted of solvent A (5:95, v / v) of methanol-water containing 0.1% ammonium hydroxide and solvent B (95:5, v / v) of methanol-water containing 0.1% ammonium hydroxide, at a flow rate of 1.0 mL / min. -1 .

[0480] Solubility of the compound in potassium phosphate buffer (pH 7.4). A stock solution (10 mM) of the test compound was prepared in DMSO and stored at -20 °C. The compound (200 µM) was incubated in 50 mM potassium phosphate buffer (pH 7.4) at 37 °C for 72 hr. The sample was vortexed, centrifuged at 12,000 rpm for 5 min, and the supernatant was filtered. Then, aliquots (100 µL) of the sample were taken and quenched by adding 200 µL of a methanol solution of 0.1% formic acid. The mixture was centrifuged again at 12,000 rpm for 5 min. The clarified supernatant was then analyzed by analytical HPLC as described. Calibration curves of the prodrug were generated by dissolving the DMSO solution of the test compound in a methanol solution of 0.1% formic acid (concentrations of 200, 100, 50, 25, 12.5, and 6.25 µM) and then analyzing by HPLC. The concentration of the soluble test compound in potassium phosphate buffer was quantified using a calibration curve. Data are expressed as mean ± SD. n = 3 separate replicates) and show representative HPLC traces.

[0481] Log using shake flask D 7.4 Assay. Aqueous potassium phosphate buffer (10 mM, pH 7.4) solutions were prepared and then saturated with n-octanol. Similarly, n-octanol was saturated with aqueous potassium phosphate buffer at pH 7.4. Both solutions were vigorously vortexed and then allowed to stand to ensure complete separation of the two phases. Shake-flask experiments were performed at room temperature. Once each phase was prepared, 1 mL of 1-octanol and 1 mL of aqueous potassium phosphate buffer were added to a glass vial. 20 µL of the test compound stock solution (10 mM stock solution in DMSO; final concentration 100 µM in the pre-saturated mixture) was added and the vial was shaken at room temperature for one hour. After one hour, the layers were separated and each aliquot (100 µL) of the n-octanol and aqueous buffer layers was treated by adding 200 µL of a methanol solution of 0.1% formic acid. The mixture was again centrifuged at 12,000 rpm for 5 minutes. The clarified supernatant was then analyzed by analytical HPLC as described. The concentration of the test compound was quantified using a calibration curve. Data are expressed as mean ± SD. n = 3 separate repetitions).

[0482] Calculate Log using the following equation. D 7.4 .

[0483] Example 2: Evaluation of the binding affinity of the conjugate The binding affinity of the compound to TTR in buffer was evaluated. Fluorescence polarization (FP) assays were used to determine the ability of AG10-L, AG10-L-E2-COOH, AG10-L-E2-naloxone, and AG10-L-E1-naloxone to displace the FP probe from TTR in buffer. Serial dilutions of the compound (0.010 µM to 20 µM) were added to 384-well plates containing the FP probe (50 nM) and TTR (300 nM) in assay buffer (PBS pH 7.4, 0.01% Triton X100, 25 μL final volume containing 1% DMSO). The samples were equilibrated by stirring on a plate shaker at room temperature for 20 min. Fluorescence polarization (excitation λ 485 nm, emission λ 525 nm, cutoff λ 515 nm) measurements were performed using a SpectraMax M5 Molecular Devices microplate reader. Fluorescence data were collected using SoftMax® Pro software version 5.4.1 (Molecular Devices, Inc.). IC50 values ​​were obtained by fitting the data to the following equation [y = (AD) / (1 +(x / C)^B) + D], where A = maximum FP signal, B = slope, and C = apparent binding constant (…). K app ), and D = minimum FP signal. Using the Cheng-Prusoff equation, according to IC 50 Value calculation combined with constant ( K d Values. Data are expressed as mean ± sd ( n = 3 separate repetitions).

[0484] The binding affinity and selectivity of the test compounds for TTR in human serum were evaluated. The binding affinity and selectivity of AG10-L, AG10-L-E2-COOH, AG10-L-E2-naloxone, and AG10-L-E1-naloxone to TTR in human serum were determined by their competitive binding with fluorescent probe exclusion (FPE) probes. AG10 and tafamidis were used as controls. Aliquots (98 μL) of human serum were mixed with 1 μL of the test compound (1.0 mM stock solution in DMSO; final serum concentration 10 μM) and 1 μL of the FPE probe (0.36 mM stock solution in DMSO; final serum concentration 3.6 μM). Fluorescence changes (λ) were monitored every 15 min at 25 °C using a SpectraMax M5 microplate reader. ex= 328 nm and λ em = 384 nm), for 6 h. Data are expressed as mean ± sd ( n = 4 separate repetitions).

[0485] Results: Fluorescence polarization (FP) was used to determine (…). K d = 394.3 nM and 258.8 nM respectively; Figure 1c The binding affinity of the prodrugs AG10-L-E1-naloxone and AG10-L-E2-naloxone was evaluated. This binding affinity is related to the binding affinity of AG10-L and AG10-L-E2-COOH (…). K d The binding affinities (262.9 nM and 213 nM, respectively) were similar. We then evaluated the TTR binding selectivity of the prodrug to >4,000 other human serum proteins using a recognized TTR serum fluorescent probe rejection (FPE) selectivity assay. Our data showed that all tested compounds maintained excellent binding selectivity to TTR in human serum (approximately 50% to 70% TTR occupancy). Figure 3d , Figure 3e The performance of AG10-L-E2-COOH (48.1% TTR occupancy) is similar to that of the TTR stabilizer chlorpromazine (an approved drug for TTR amyloidosis; 49.2% TTR occupancy).

[0486] Example 3: Stability Analysis Stability determination of the test compound in human and rat plasma. The test compound (5 μM) was incubated in human or rat plasma at 37 °C. 100 μL plasma samples were analyzed at 0 hr, 0.25 hr, 2 hr, 4 hr, and 24 hr. All samples were processed by adding 200 μL of quenching buffer (0.1% formic acid in methanol) followed by centrifugation at 15,000 rpm for 5 min. The supernatant was incubated at -20 °C for 5 min and then centrifuged again at 15,000 rpm for 5 min. The supernatant was analyzed by analytical HPLC as described. Data are expressed as mean ± SD ( n = 3 separate repetitions).

[0487] Results: We first evaluated the stability of the prodrug in human plasma. AG10-L-E1-naloxone was unstable (0% remaining after 1 min of serum incubation). Figure 1fThis aligns with the instability of alkyl-based ester prodrugs. In contrast, PABA-based esters (AG10-L-E2-naloxone and C12-E2-naloxone) are completely protected from hydrolysis in human plasma. While aryl-based esters are more stable than their alkyl counterparts, we are still surprised by the complete stability of these prodrugs in human plasma. Our modeling studies show that the linker we used is long enough to extend the PABA ester in the TTR T4 binding bag, thereby allowing, in principle, plasma esterases to target our prodrug, even if it is bound to the TTR. Figure 2b To investigate these findings, we evaluated the stability of the prodrug in rat serum. As expected, the alkyl ester prodrug (AG10-L-E1-naloxone) was hydrolyzed most rapidly (remaining at 0% after 1 min of serum incubation). Figure 1g The PABA ester AG10-L-E2-naloxone exhibited high stability (60% remaining after 15 min and 3.8% remaining after 2 hr). The hydrolysis rate of C12-E2-naloxone (34.2% remaining after 15 min and 0% remaining after 2 hr) was slightly faster than that of AG10-L1-E2-naloxone. The difference in prodrug hydrolysis between human and rat plasma points to carboxylesterases 1 and 2 (CES1 and CES2), which are present in rat plasma but not in human plasma.

[0488] Docking with TTR and CES2. Computer simulations of mouse Ces2c docking with AG10-L-E2-naloxone were performed using Dock6. The crystal structures of TTR (pdb id: 4HIQ) and Ces2c (pdb id: 8AXC) were obtained from the RCSB.org protein database website. The geometry of AG10-L-E2-naloxone was optimized at the B3LYP / 6-311G* level using Gaussian09. For docking with TTR and Ces2c, AG10-L-E2-naloxone was split into two: AG10-connector and naloxone-connector. The geometry of both ligands with shorter connectors was optimized at the same B3LYP / 6-311G* level. Molden was used to analyze the ligand geometry and prepare ligand files for docking. The UCSF Chimer program was used for receptor docking preparation and analysis of docking results. AG10-L A and L B -E2-naloxone was docked to TTR and Ces2c, respectively. TTR--- AG10-L A and Ces2c---L BA docking complex of -E2-naloxone was stacked on an optimized structure of AG10-L-E2-naloxone to observe the possibility of forming a ternary complex of TTR-AG10-L-E2-naloxone-Ces2c.

[0489] Example 4: Stability of the conjugate to carboxylesterase hydrolysis Materials and methods Metabolic studies of AG10-L-E2-naloxone were conducted using recombinant human carboxylesterase. The naloxone was synthesized from CES1 (30 µg / mL). -1 Or 100 µg mL -1 ) or CES2 (30 µg mL -1 The reaction mixture, composed of 50 mM potassium phosphate buffer, was incubated at 30 °C for 5 min. The reaction was initiated by adding 5 µM AG10-L-E2-naloxone (or DMSO as a negative control). After incubation, 100 μL of sample from the reaction mixture was quenched at 0 hr and 1 hr by adding 100 μL of quenching buffer (0.1% formic acid in methanol) to the sample to terminate the reaction at each specified time point. The sample was then centrifuged at 12,000 rpm for 5 min at 4 °C. The supernatant (150 μL) was further diluted by adding 75 μL of 0.1% formic acid aqueous solution. The sample was stored at -20 °C until analyzed by HPLC as described above. Data are expressed as mean ± SD. n = 3 separate repetitions).

[0490] Microsomal stability assay. 2 µL of AG10-L-E2-naloxone stock solution (1 mM in DMSO) was added to a microcentrifuge tube containing 3.2 µL acetonitrile and 194.8 µL potassium phosphate buffer (50 mM, pH 7.4, containing 3 mM MgCl2). The tube was pre-incubated at 37°C for 10 min, and then liver microsomes were added. Human, monkey, or rat liver microsomes and NADPH regeneration solution (NADP, glucose-6-phosphate, and glucose-6-phosphate dehydrogenase; concentration 2 mg / mL) were added. -1 And respectively 2 mM, 10 mM and 2 units mL -1Add to potassium phosphate buffer (water is used instead of NADPH in the negative control). Preheat the reaction mixture at 37°C for 10 min. NADPH is generated using the RapidSTART NADPH regeneration system (Xenotech) by adding 0.7 mL of high-purity water to the frozen component in the RapidStart vial, vortexing, and then adding the activated RapidStart solution to the incubator. Add 200 µL of this NADPH regeneration solution to each pre-incubated assay tube containing 200 µL of AG10-L-E2-naloxone in phosphate buffer. The final reaction concentrations of AG10-L-E2-naloxone, microsomes, NADP, glucose-6-phosphate, and glucose-6-phosphate dehydrogenase are 5 µM, 1 mg / mL, and 1 mg / mL, respectively. -1 1 mM, 5 mM and 1 unit mL -1 Incubate the vials at 37°C. At 0 hr and 1 hr, remove 100 µL aliquots and add 100 µL of quenching buffer (containing 15 ng / mL) to each aliquot. -1 The microsomal reaction was terminated with naloxone-D5 as an internal standard in a 0.1% formic acid methanol solution. The sample was centrifuged at 12,000 rpm for 5 min and the supernatant was further diluted (150 µL) by adding 75 µL of 0.1% formic acid aqueous solution. The sample was stored at -20 °C until analysis using a validated LC-MS / MS method with a triple quadrupole mass spectrometer (QQQ). Data are expressed as mean ± SD. n = 3 separate repetitions).

[0491] Microsomal stability assays were performed using loperamide or digitalis saponins. The microsomal assays described above were also performed in HLM in the presence of loperamide or digitalis saponins (final concentration 100 µM). Loperamide or digitalis saponins were added to the microsomal buffer before being added to the test compound.

[0492] Stability assay of AG10-L-E2-naloxone in rat plasma in the presence of CES inhibitors. Rat plasma was pre-incubated at room temperature for 5 min with either the CES2 inhibitor loperamide (100 µM) or the CES1 inhibitor digitoxin (100 µM) or DMSO (negative control). Then, AG10-L-E2-naloxone (5 μM) was added to the incubation mixture. Samples were analyzed at 0 hr, 0.5 hr, and 1 hr. All samples were processed by adding 200 μL of quenching buffer (0.1% formic acid in methanol) followed by centrifugation at 15,000 rpm for 5 min. The supernatant was incubated at -20°C for 5 min and then centrifuged again at 15,000 rpm for 5 min. The supernatant was analyzed by analytical HPLC as described. Data are expressed as mean ± SD. n = 3 separate repetitions).

[0493] Stability determination of AG10-L-E2-naloxone in monkey and rabbit plasma. AG10-L-E2-naloxone (5 μM) was incubated in monkey or rabbit plasma at 37 °C. 100 μL samples were analyzed at 0 hr and 24 hr. All samples were processed by adding 200 μL of quenching buffer (0.1% formic acid in methanol) followed by centrifugation at 15,000 rpm for 5 min. The supernatant was incubated at -20 °C for 5 min and then centrifuged again at 15,000 rpm for 5 min. The supernatant was analyzed by analytical HPLC as described. Data are expressed as mean ± SD ( n = 3 separate repetitions).

[0494] result Carboxylesterases (CESs) play a crucial role in the hydrolytic inactivation of many clinically used ester drugs and the bioactivation of prodrugs. CES1 and CES2 are the two most prevalent forms of CES in humans. Human CES1 plays a major role in the bioactivation of many prodrugs, while CES2 plays a minor role. Significant species differences exist in the tissue distribution and catalytic activity of CESs, making human translation challenging. CES1 is highly expressed in the liver, while CES2 is mainly found in the small intestine, but is also present in the colon, kidney, and liver (at protein levels 10-15 times lower than CES1).

[0495] We investigated the potential role of CES1 / CES2 in the bioactivation of AG10-L-E2-naloxone. C12-E2-naloxone is insoluble in buffer solution. Figure 1bThis prevented us from evaluating its CES selectivity. Our in vitro data on purified human CES clearly demonstrate that AG10-L-E2-naloxone is a substrate specific to CES2. Figure 2c ). AG10-L-E2-Naloxone CES2 Specificity Completely unexpected, because only a few prodrugs (such as procaine and irinotecan) were involved. selectively Procaine is bioactivated by CES2 (which is also bioactivated by CES1 and other plasma proteins). For example, in addition to hepatic CES2, procaine undergoes rapid hydrolysis by butyrylcholinesterase in human plasma. In contrast, CES2 bioactivates irinotecan extremely slowly (less than 2% of the dose is bioactivated by CES2). To further investigate CES2 specificity and evaluate potential human translation, we evaluated the metabolic bioactivation of AG10-L-E2-naloxone in liver microsomes from humans, rats, and monkeys. Figure 2d Human liver microsomes (HLMs) efficiently produce naloxone from AG10-L-E2-naloxone. The selective CES2 inhibitor loperamide significantly reduced naloxone release, while the CES1 inhibitor digitalis saponin had no effect. The amount of naloxone released from AG10-L-E2-naloxone in rat liver microsomes (RLMs) was lower than that formed in HLMs, primarily due to higher metabolic activity (via CYP dealkylation) of naloxone released from RLMs compared to HLMs. Since CES enzyme expression is similar in monkeys and humans, we evaluated the bioactivation of AG10-L-E2-naloxone in monkey liver microsomes (MLMs). The amount of naloxone released from MLMs was similar to that from HLMs, suggesting that monkeys are a suitable animal model for evaluating AG10-L-E2-naloxone.

[0496] The specificity of CES2 for the bioactivation of AG10-L-E2-naloxone was also confirmed by measuring the plasma stability of CES1 and CES2 inhibitors in rats. Figure 2e We also confirmed our results by evaluating the stability of AG10-L-E2-naloxone in monkey plasma (plasma without CES1 or CES2; stability of 100% after 24 hr) and rabbit plasma (plasma with high abundance of CES1 and CES2; 0% remaining after 24 hr).

[0497] Although the crystal structure of human CES1 is available, the first crystal structure of mouse CES2 has recently been reported as a potential ortholog of human CES2. The overall structure of mouse CES2 is similar to that of human CES1, differing mainly in the proximity of the active site. We conducted docking studies of AG10-L-E2-naloxone with both human CES1 and mouse CES2. AG10-L-E2-naloxone did not bind to CES1. Conversely, AG10-L-E2-naloxone showed a highly compatible active site with CES2, and the nucleophilic serine (S230) is adjacent to the carbonyl group of the ester group in AG10-L-E2-naloxone. Figure 2b This data represents the first docking experiment between a specific CES2 substrate and the CES2 enzyme. Insights into the binding mode and selectivity of CES2 to AG10-L-E2-naloxone can help in the design of additional CES2-specific prodrugs. The lower concentration of hepatic CES2 compared to CES1 provides unique CES2 bioactivation for AG10-L-E2-naloxone. This could allow us to control the slow and continuous release of naloxone from AG10-L-E2-naloxone (discussed below in the monkey study in Example 10).

[0498] Example 5: Subcutaneous administration of AG10-L-E2-naloxone to rats resulted in a prolonged and controlled release of naloxone. There was no dramatic increase in plasma and brain concentration. Materials and methods Evaluation of the subcutaneous and intravenous pharmacokinetic profiles of AG10-L-E2-naloxone and naloxone in rats. Subcutaneous (sc) and intravenous (iv) pharmacokinetic (PK) studies were performed using male Sporgodory rats (225–250 g; 7–8 weeks old) cannulated from the Charles River. For the intravenous PK studies, each animal received a single intravenous bolus dose of either naloxone or AG10-L-E2-naloxone (60 µmol kg) in 500 µL of dosing solution (10% DMSO, 20% PEG, 70% H2O). -1 Equivalent to 20 mg kg -1 Naloxone was administered, followed by an injection of 200 µL of sterile saline to flush the jugular vein cannula (n = 3 rats per group). At predetermined time points (0.033, 0.25, 0.5, 1, 2, 4, 6, 8, 12, and 24 hours after administration), 200 µL of blood samples were collected from each rat via the jugular vein cannula into heparinized tubes, and the volume was replaced with sterile standard saline. For subcutaneous pk studies, 15 µmol / kg naloxone was administered to the nape of the neck region. -1 or 60 µmol kg -1 AG10-L-E2-naloxone 15 µmol kg -1 or 60 µmol kg-1 500 µL of the drug solution (per 250 g rat) was administered. In each of the four groups (n = 3 rats per group), blood samples were collected from each rat via jugular vein cannulation into heparinized tubes at predetermined time points (0.033, 0.25, 0.5, 1, 2, 4, 6, 8, 12, 24, 48, 72, and 96 h post-administration), and the same volume was replaced with sterile standard saline. Plasma samples were prepared directly after blood collection by centrifugation at 1,500 g for 10 min at 4 °C. Next, aliquots of 50 µL plasma samples were precipitated with 2 volumes of extraction buffer (a methanol solution of 0.1% formic acid containing 15 ng / mL naloxone-D5). The samples were vortexed for 30 sec and then held at -20 °C for 5 min. The samples were then centrifuged at 15,000 rpm for 5 min; the supernatant was collected and kept in a -20°C freezer for 5 min, followed by another centrifugation at 15,000 rpm for 5 min. On the same day of the study, the samples were analyzed using triple quadrupole mass spectrometry (QQQ) with LC-MS / MS to quantify the concentrations of naloxone and AG10-L-E2-naloxone in all groups.

[0499] Sample preparation and LC-MS / MS analysis of the experimental compounds. Sample preparation. Stock solutions of naloxone (20 mM) and AG10-L-E2-naloxone (10 mM) were prepared in DMSO solution. Working solutions were prepared by serial dilution of the DMSO solutions mentioned above. Calibration curve standards were prepared by incorporating aliquots (1 µL) of each working solution into 49 µL of rat plasma from the blank control group. Quality control (QC) samples prepared at low, medium, and high concentrations were extracted simultaneously with animal samples. Extraction was performed using 2 volumes of extraction buffer (containing 15 ng / mL). -1 Naloxone-D5 was precipitated in 50 µL aliquots of plasma using a methanol solution of 0.1% formic acid. The corresponding double blank samples were extracted with extraction buffer (i.e., methanol solution of 0.1% formic acid) without internal standards. Samples were processed in the same manner described above. To quantify naloxone, 20 µL of sample was injected for LC-MS / MS analysis. Analytes in plasma samples were quantified using a triple quadrupole mass spectrometer (QQQ). The mobile phase consisted of solvent A (0.1% formic acid aqueous solution containing 0.5 mM ammonium formate) and solvent B (methanol solution of 0.1% formic acid) at a flow rate of 0.5 mL / min. -1LC-MS / MS analysis was performed on a Waters™ XBridge C18 (4.6 x 150 mm, 5 µm). The turbine spray ionization source was set to positive ionization mode. The test compounds were identified and quantified using fragmented patterns and peak areas, respectively. LC-MS / MS data were collected from Agilent Masshunter software. Naloxone-D5 was used as an internal standard. Gas flow rate, sprayer, sheath gas temperature, sheath gas flow rate, capillary voltage, nozzle voltage, and gas temperature were set to 7 l / min, 30 psi, 250 °C, 12 l / min, 3500 V, 500 V, and 300 °C, respectively. LC-MS / MS analysis was performed using the following gradient method: linear increase from 0% to 100% solvent B over 0–3 min, holding at 100% solvent B for 5.5 min, then decreasing from 100% to 0% solvent B over 0.5 min, and finally holding at 0% solvent B for 1 min. The retention times for naloxone and naloxone-D5 were 4.62 and 4.62 min, respectively. The total run time was 7 min. The mean ± sd concentration of naloxone was determined based on a calibration curve of naloxone and internal standard in plasma generated by an LC-MS / MS analyst. Samples were diluted when the concentration exceeded the calibration curve range. The table below lists the detailed mass spectrometer conditions for multiple reaction monitoring (MRM) for each compound. The limit of quantitation (LLOQ) for naloxone was 1 nM. The linear range was calculated based on regression analysis between the analyte / internal standard peak area ratio and the analyte concentration. In each case, the accuracy was between 80% and 120%.

[0500] Evaluation of AG10-L-E2-naloxone and naloxone brain uptake in rats. This study was conducted using male Spögdolly rats with jugular vein cannulation. Animals were randomly assigned to four treatment groups (each group... n = 3 animals). Each animal received a single subcutaneous dose of 15 µmol kg. -1 Naloxone or AG10-L-E2-naloxone was administered in 500 µL of dosing solution (10% DMSO, 20% PEG400, and 70% sterile deionized water). 90 mg / kg was injected intraperitoneally at 10 and 30 min after dosing. -1 Chlorobenzophenone and 9 mg kg -1 Rats were anesthetized with xylazine. CSF samples were collected from the cerebellomedullary cistern using a 22-gauge needle. Blood was collected by phlebotomy from the aorta using a 20-gauge needle, followed by decapitation and brain collection. Sample preparation and LC-MS / MS analysis of experimental compounds AnalysisSample Preparation. Immediately freeze the brain in liquid nitrogen. Process and analyze plasma samples in the same manner as plasma samples described in pharmacokinetic studies. Store CSF and brain samples at -80°C for further analysis. Prepare stock solutions of naloxone (20 mM) and AG10-E2-naloxone (10 mM) in DMSO solution. Prepare working solutions by serially diluting the DMSO solution mentioned above. Prepare calibration curve standards by incorporating aliquots (1 µL) of each working solution into 49 µL of artificial CSF (125 mM NaCl, 2.5 mM KCl, 1.26 mM CaCl2, and 1.18 mM MgCl2) or blank rat brain homogenate. Simultaneously extract animal samples with quality control (QC) samples prepared at low, medium, and high concentrations. Prepare brain homogenates by homogenizing brain tissue with 2 volumes (w:v) of homogenization solution (PBS buffer). Use 2 volumes of extraction buffer (containing 0.1% formic acid and 15 ng / mL) -1 Naloxone-D5 was precipitated in 50 µL aliquots of brain homogenate or CSF sample using methanol-water (95:5, v / v). The corresponding double blank samples were extracted with extraction buffer (i.e., methanol-water (95:5, v / v) containing 0.1% formic acid) without an internal standard. The CSF and brain homogenate samples were vortexed for 30 seconds and then held at -20°C for 5 minutes. The samples were then centrifuged at 15,000 rpm for 5 minutes; the supernatant was collected and held at -20°C for 5 minutes and centrifuged again at 15,000 rpm for 5 minutes. The samples were stored at -80°C for further analysis. LC-MS / MS analysis was also performed as described in the pharmacokinetic studies of these molecules. The LLOQ of naloxone and AG10-L-E2-naloxone in brain and CSF matrices was 1 nM. Brain (ng g) -1 ) relative to plasma concentration (ng / mL) -1 The ratio is expressed as a percentage of brain / plasma ratio. CSF (ng / mL) -1 ) relative to plasma concentration (ng / mL) -1 The ratio is expressed as a percentage of the CSF / plasma ratio.

[0501] Rats receiving LD 50 Survival rates following doses of fentanyl or morphine. Male Sporgodory rats (225–250 g; 7–8 weeks old) were obtained from Charles River Laboratories. Rats were randomly assigned to three groups (carrier, naloxone, and AG10-L-E2-naloxone). Rats initially received a subcutaneous dose of 100 mg / kg. -1 Morphine (per group) n = 9Five minutes after the morphine dose, rats were administered a single subcutaneous dose of the carrier (10% DMSO, 20% PEG400, and 70% H2O) and naloxone (15 µmol kg). -1 Equivalent to 5 mg kg -1 ) or AG10-L-E2-naloxone (15 µmol kg -1 Equivalent to 15.8 mg / kg -1 Naloxone and AG10-L-E2-naloxone were dissolved in the same mediator administration solution. Morphine was dissolved in water. The injection volumes of morphine and the antagonist were 200 and 500 µL, respectively. The rats were then returned to their cages under standard conditions. The number of dead rats and their approximate time of death were recorded. Six hours after the first morphine dose, all surviving rats received a second subcutaneous dose of 50 mg / kg. -1 Morphine. Return the rats to their cages and observe to record any deaths. Record the total number of dead rats and calculate the survival rate for each group.

[0502] In the fentanyl mortality study, rats were first given a subcutaneous dose of 3 mg / kg. -1 Fentanyl (per group) n = 8 Five minutes after the fentanyl dose, rats were administered a single subcutaneous dose of the carrier (10% DMSO, 20% PEG400, and 70% H2O) and naloxone (15 µmol / kg). -1 Equivalent to 5 mg kg -1 ) or AG10-L-E2-naloxone (15 µmol kg -1 Equivalent to 15.8 mg kg -1 Naloxone and AG10-L-E2-naloxone were dissolved in the same mediator administration solution. Fentanyl was dissolved in water. The injection volumes of fentanyl and the antagonist were 200 µL and 500 µL, respectively. The rats were then returned to their cages under standard conditions. The number of dead rats and their approximate time of death were recorded. Six hours after the first morphine dose, all surviving rats received a second subcutaneous dose of 3 mg / kg. -1 Fentanyl. Return the rats to their cages and observe to record any deaths. Record the total number of dead rats and calculate the survival rate for each group.

[0503] Reversal and prevention of fentanyl-induced respiratory depression in rats. The efficacy of antagonists in reversing fentanyl-induced respiratory depression in rats was evaluated using a whole-body plethysmography device from DSI Buxco (St. Paul, MN; equipped with FinePointe software). Sporgodori rats (225–250 g; 7–8 weeks old) (each group) n = 6Rats were acclimatized for 30 min in a silencing, enclosed plethysmography chamber under a continuous fresh air flow (2.2 L / min). Rats were awake and allowed free movement within the test chamber during the monitoring period. For baseline measurements, rat respiration was recorded for 20 min prior to injection. Rats were then removed from the test chamber and sequentially administered fentanyl (3 mg / kg). -1 A single subcutaneous or intravenous dose), followed by a single subcutaneous or intravenous dose of the antagonist (15 µmol kg) one minute later. -1 or 60 µmol kg -1 Treatment: Rats were returned to the test chamber, and respiratory parameters were measured for 90 min. Six and 24 hours after antagonist administration, rats were given two additional subcutaneous or intravenous doses of fentanyl (1.5 mg kg each). -1 Rats were attacked with fentanyl. Respiratory parameters were measured for 90 minutes after each fentanyl injection. Ventilation was measured by recording changes in ventricular pressure, and the data were then processed and digitized. The software calculated the mean tidal volume (V) for each rat. T (ml / breath), respiratory rate (f: breaths / minute), and minute ventilation (V) E = V T * f: ml / min). Since the minute ventilation is V... T The product of f and therefore reported is considered a scientifically relevant and quantifiable net physiological mean. Baseline minute ventilation data for each rat were averaged and designated as 100% of the baseline value. The minute ventilation readings for each rat after administration were then normalized and reported at 5-minute intervals based on the following equation: Minute ventilation (percentage relative to control baseline) = Minute ventilation at a specific time / Baseline minute ventilation * 100.

[0504] To administer an antagonist prophylactically to counteract fentanyl-induced respiratory depression: rats (each group) n = 6 Rats were acclimatized to the plethysmography chamber for 30 min, and baseline measurements were recorded as described above. Rats received a single subcutaneous dose of the mediator and naloxone (15 µmol / kg). -1 and 60 µmol kg -1 ) or AG10-L-E2-naloxone (60 µmol kg -1 ), and respiratory parameters were measured for 90 minutes. Six hours later, a single subcutaneous dose of fentanyl (1.5 mg / kg) was administered. -1 Rats were attacked and returned to the plethysmography chamber. Respiratory parameters were measured for 90 min, and data were processed and reported as described above. Treatment doses of morphine and fentanyl were determined in preliminary studies and also based on published literature. The dose of morphine used (100 mg / kg)-1 The reported LD50 of morphine in rats is 64-140 mg / kg. -1 For fentanyl, we use a reported value of 3 mg / kg. -1 These dosages were confirmed in preliminary experiments in which we started with low doses of morphine and fentanyl and no fatalities were observed. Overdose studies of morphine and fentanyl were approved by the University of the Pacific (IACUC).

[0505] Preliminary evaluation of AG10-L-E2-naloxone toxicity in rats. The in vivo toxicity of AG10-L-E2-naloxone in rats was evaluated. Adult male Charles River Sporgori rats weighing 225–250 g were used in the study. Three groups of rats were used. The first group was treated with 500 µL of a medium (10% DMSO, 20% PEG, and 70% H2O). The second group was treated with AG10-L-E2-naloxone dissolved in 500 µL of the medium (60 µmol / kg). -1 The third group was treated with fentanyl (3 mg / kg). -1 Treatment with a subcutaneous dose, followed by AG10-L-E2-naloxone (60 µmol kg) -1 Treatment was initiated, followed by two doses of fentanyl (1.5 mg kg) at 6 hours and 24 hours. -1 Treatment was administered subcutaneously each time. Animal weight was monitored weekly for four weeks. Weight is expressed as mean ± sd (per group). n = 6 biological replicates).

[0506] result AG10-L1-E2-Naloxone for the prevention of death in rats following two doses of morphine or fentanyl We evaluated the pharmacokinetic (PK) properties of naloxone and AG10-L-E2-naloxone in rats. Figure 3a Two compounds (15 µmol kg) were used. -1 Equivalent to 5 mg kg -1 The naloxone dose was administered to rats as a single subcutaneous dose, and blood samples were drawn from the jugular vein cannula at predetermined time points (within the range of 5 min to 72 hr). Figure 3a Consistent with our hypothesis, the PK profile of AG10-L-E2-naloxone differed significantly from that of naloxone. Both compounds were rapidly absorbed (naloxone was detected in plasma 2 min after administration of either free naloxone or AG10-L-E2-naloxone) and T... maxThe time is approximately 15 minutes. The C of naloxone administered in free form... max (1360 nM) significantly higher than the C of naloxone released from AG10-L-E2-naloxone. max (16.9 ± 2.1 nM). However, although no measurable amount of free naloxone was observed 4 hours after administration, AG10-L-E2-naloxone and its released naloxone remained even after 72 hours. Figure 3a The PK profile of naloxone showed a typical spike curve (ranging from approximately 1360 nM at 15 min). max C at 34 nM at 4 hr min Surprisingly, the release of naloxone from AG10-L-E2-naloxone follows a flatter curve (ranging from approximately 17 nM C at 15 min). max C at 2.4 nM at 72 hours min We believe this will be superior to the early concentration spikes observed with naloxone (which may cause withdrawal symptoms). Importantly, human data show that naloxone has a lower C-level concentration after subcutaneous administration. max (3.5 nM for a 2 mg dose and 17 nM for a 5 mg dose, which was verified for IM or SC doses) approximately 10 nM. To the best of our knowledge, this is the first PK profile instance demonstrating a smooth release of a therapeutic molecule without the presence of typical peaks and troughs or burst effects observed on multiple runs. This data strongly supports and validates our novel approach, in which the hydrophilicity of our prodrug controls the release of these molecules from subcutaneous tissue. This combination of slow release and TTR binding results in a smooth and stable plasma concentration of naloxone released in rats.

[0507] Then, we evaluated the blood-brain barrier (BBB) ​​permeability of AG10-L-E2-naloxone (5 mg / kg; subcutaneous) in rats. Figure 3b The concentrations of naloxone released from AG10-L1-E2-naloxone in the brain (7.8 nM and 30.1 nM after 10 and 30 min, respectively) were approximately 30 to 60 times lower than the concentrations of naloxone administered in free form (236.3 nM and 1721.7 nM after 10 and 30 min, respectively). Similar data were observed for the two compounds in cerebrospinal fluid (CSF). Figure 3cThis clearly shows that a significant amount of naloxone released from AG10-L1-E2-naloxone crosses the BBB. No detectable concentrations of parent AG10-L1-E2-naloxone were found in the brain or CSF. These data support our hypothesis that hydrophilicity and selective TTR binding limit the ability of the AG10-conjugate to cross the BBB. The brain / plasma ratio of naloxone released from AG10-L1-E2-naloxone (250.3% after 30 min) and the CSF / plasma ratio (173.3% after 30 min) were remarkably similar to the percentage of naloxone administered in free form. This is significantly higher than 2% (the BBB penetration cutoff; molecules with plasma concentrations <2% in the brain or CSF have been determined not to cross the BBB).

[0508] We evaluated the potential of our prodrug as an antidote for saving lives from opioid overdose. Two opioids were evaluated: morphine and fentanyl. Although the half-lives of fentanyl (approximately 7 hours) and morphine (2 to 3 hours) were... t 1 / 2 The duration of overdose is relatively short, but the duration of overdose can last up to 24 hours for fentanyl and up to 120 hours for morphine (depending on the usually unknown opioid dose). Treatment doses for morphine and fentanyl were determined in preliminary studies and also based on published literature. The dosage of morphine (100 mg / kg) -1 Reported LD50 of morphine in rats 50 64-140 mg kg -1 For fentanyl, we use a reported value of 3 mg / kg. -1 Three groups of rats (each group) n = 9 ) Administer morphine (100 mg / kg) -1 (Single subcutaneous dose). 5 min later: Group 1 received the medium; Group 2 received naloxone (15 µmol kg). -1 Equivalent to 5 mg kg -1 (single subcutaneous dose), and group 3 received a molar equivalent dose of AG10-L-E2-naloxone (15 µmol kg). -1 Equivalent to 15.8 mg kg -1 (Single subcutaneous dose). 15 µmol kg of the selected naloxone and AG10-L-E2-naloxone. -1 The dose is higher than the dose used in the literature (1 mg kg). -1 Equivalent to 3 µmol kg -1This ensures that respiratory depression from morphine and fentanyl doses is almost completely reversed by naloxone, which is released more slowly from our prodrug. All mediator-treated control groups died after the first morphine dose, while the naloxone and AG10-L-E2-naloxone groups remained alive. Figure 3d Six hours later, administer the second dose of morphine (50 mg / kg). -1 A single subcutaneous dose was administered to animals treated with naloxone and AG10-L-E2-naloxone to simulate re-anesthesia. All animals in the naloxone group died within 2 hours of the second morphine dose. Notably, all animals in the AG10-L1-E2-naloxone group survived and remained alive for several weeks after administration.

[0509] We also used two doses of fentanyl (3 mg kg each) -1 Subcutaneous, at times = 0 hr and 6 hr; each group n = 8 Attacking rats to conduct similar experiments Figure 3e Following the first dose of fentanyl, six rats in the mediator group died, and the remaining rats died after the second dose. In the naloxone group, all eight rats died after the second dose of fentanyl. Importantly, after the second dose of fentanyl, only one of the eight rats in the AG10-L-E2-naloxone treatment group died. Figure 3e The data clearly demonstrate that AG10-L1-E2-naloxone is more effective than naloxone in reducing mortality in rats administered two high doses of fentanyl. Given the urgent need to develop an effective antidote for fentanyl, the following studies will focus on evaluating our prodrug against fentanyl overdose.

[0510] Reversal of Opioid-Induced Respiratory Depression (OIRD) in Rats Systemic side effects of opioid toxicity include life-threatening intraepithelial neoplasia (OIRD), which can have potentially fatal consequences for patients. OIRD is also a major obstacle to the safe administration of morphine for acute pain following injury or trauma during emergency situations. OIRD can be properly managed when monitored by trained medical personnel and when appropriate equipment is available. However, this is difficult to achieve in emergency situations where these options are unavailable, such as battlefield environments or weaponized synthetic opioids. Although long-term administration can lead to tolerance to respiratory depression, it can also cause death in opioid-naïve patients, meaning it is one of the most concerning side effects for clinicians in the postoperative setting. We tested our prodrug (15 µmol kg) using a non-invasive whole-body plethysmography method. -1 It is equivalent to 5 mg kg -1 Naloxone; single subcutaneous dose, n = 6) Reversing high-dose fentanyl (LDL) 50 3 mg / kg-1 The ability of OIRD (single subcutaneous dose) to ( Figures 3f to 3i ).

[0511] After the first fentanyl dose ( Figure 3g No reversal of OIRD was observed with subcutaneous administration mediators or lipotropic naloxone prodrugs containing PABA ester C12-E2-naloxone (although hydrolyzed in rat serum); Figure 1g This was predictable because lipogenesis slowed the release of the conjugate from the subcutaneous tissue. We also had difficulty formulating C12-E2-naloxone into the dosing medium, requiring doubling the volume of the dosing medium. Two out of six rats in both the medium-treated and C12-E2-naloxone-treated groups died after the first fentanyl dose, so we did not continue testing the C12-E2-naloxone prodrug. As expected, naloxone and the alkyl ester AG10-L-E1-naloxone provided rapid reversal of OIRD after 10 min (confirmed by a rapid recovery of minute ventilation). Figure 3g For AG10-L-E2-naloxone, the minute ventilation in rats gradually recovered after administration, but did not reach the level close to that of normal rats (the minute ventilation at 90 min was 52.5 ± 4.5% of the control).

[0512] To test the ability of the prodrug to prevent fentanyl-induced respiratory depression (i.e., a state similar to re-anesthesia) after initial reversal, a second dose of fentanyl (1.5 mg / kg) was administered 6 hours after naloxone or the prodrug. -1 A single subcutaneous dose was used to attack rats ( Figure 3h As expected, naloxone did not effectively reverse OIRD, and four out of six rats in the naloxone group died after the second fentanyl dose. AG10-L-E1-naloxone was also ineffective (as confirmed by the stability of rat serum showing rapid hydrolysis of the alkyl ester). Figure 1g AG10-L-E2-naloxone is the only prodrug that showed significant OIRD reversal 6 hours after the second dose of fentanyl. Figure 3h 24 hours after administration of naloxone or the prodrug, administer the third dose of fentanyl (1.5 mg / kg). -1 A single subcutaneous dose was administered to attack rats. This again demonstrated that AG10-L-E2-naloxone was more effective than naloxone and AG10-L-E1-naloxone (one of six rats in the AG10-L-E1-naloxone group died after the third fentanyl dose) in preventing death and reversing OIRD. Figure 3iThe slow onset of action of AG10-L-E2-naloxone suggests that the initial concentration of naloxone released from the prodrug needs to be increased to achieve effective salvage from the first fentanyl dose. However, this slow release profile may be helpful for certain therapeutic applications, such as the treatment of analgesia and neonatal conditions (discussed below).

[0513] Example 6: AG10-L1-E2-Naloxone is more effective than naloxone and nalmefone in preventing fentanyl re-anesthesia. efficient Our data show that low-dose AG10-L-E2-naloxone effectively reduced fentanyl levels after two doses (3 mg kg each). -1 The mortality rate afterward. However, the plethysmography results ( Figure 3g This indicates a need to increase the initial concentration of naloxone released. Therefore, we evaluated a higher dose of AG10-L-E2-naloxone (4-fold higher; 60 µmol kg). -1 Equivalent to 63mg / kg -1 Pharmacokinetics overview of ) Figure 4a For this extremely high dose, the C of naloxone administered in free form... max (6498 ± 2.15 nM) was significantly higher than the C of naloxone released from AG10-L-E2-naloxone. max (approximately 30 nM) Figure 4a In contrast to naloxone, and as observed for lower doses, the release of naloxone from AG10-L-E2-naloxone follows a similarly flatter curve (ranging from approximately 30 nM C at 30 min). max C at 15 nM at 96 hr min (96 hr is very good for supporting monkeys). We then tested higher doses of AG10-L-E2-naloxone (60 µmol kg). -1 Equivalent to 63 mg kg -1 The efficacy of ) Figures 4b to 4e We also included extremely high doses of two FDA-approved antidotes, naloxone (60 µmol kg). -1 Equivalent to 20 mg kg -1 ) and nalmefene (60 µmol kg -1 Equivalent to 21 mg kg -1 As a control, nalmefene effectively reversed OIRD within 2 minutes after the first fentanyl dose (consistent with the reported rapid absorption; T...). max (Approximately 20 minutes) Figure 4cHowever, administration of nalmefone to rats resulted in significant hyperventilation and some spasms or seizures, indicating the deposition of opioid withdrawal symptoms. Three rats in the fentanyl control group died. Higher doses of AG10-L-E2-naloxone (60 µmol / kg) were also observed. -1 The efficacy of the (reversible within 20 min) dose was superior to that of the lower dose (15 µmol kg). -1 (Reversed within 50 minutes) Figure 4c Following a second fentanyl attack on rats, nalmefene and AG10-L-E2-naloxone were equally effective in reversing OIRD, indicating the presence of effective levels of naloxone release 6 hours after administration of AG10-L-E2-naloxone. Figure 4d Naloxone control rats did not effectively reverse OIRD after administration of a second dose of fentanyl (i.e., 6 hours after naloxone administration; 2 rats in the naloxone control group died). We then challenged the rats with a third dose of fentanyl 24 hours after administration of the antagonist (to simulate re-anesthesia). Figure 4e It is noteworthy that while nalmefene did not effectively reverse fentanyl-induced OIRD, AG10-L-E2-naloxone effectively counteracted OIRD. This data demonstrates that we have developed what we consider to be the most potent μ-opioid antagonist prodrug (effective as a reversal agent and potentially an ideal prophylactic agent).

[0514] Example 7: Compared to lower doses, higher doses of naloxone did not provide additional efficacy. Surprisingly, there was a lack of correlation between the pharmacokinetics and pharmacodynamic profile of naloxone. Pharmacokinetic data showed a considerably high plasma concentration of free naloxone (25 nM) at 6 hr, similar to the concentration of naloxone released from AG10-L-E2-naloxone (21 nM). Figure 4a However, rats treated with naloxone performed poorly in plethysmography after a second fentanyl dose, and two rats died despite sufficient naloxone in circulation. Figure 4a and Figure 4d This data suggests that the initial extremely high levels of naloxone (6498 nM at 15 min) may actually be harmful and make animals more susceptible to the toxic effects of a second dose of fentanyl compared to the low sustained levels of naloxone from AG10-L-E2-naloxone (concentration at 15 min = 20 nM).

[0515] To investigate this, we conducted another experiment in which fentanyl and its antagonist (naloxone and AG10-L-E2-naloxone) were administered intravenously at a concentration of 60 µmol kg. -1 Equivalent to 20 mg kg -1(single intravenous dose) Figure 4f Pharmacokinetic data showed that naloxone (concentration at 15 min = 20597 nM) and naloxone versus AG10-L-E2-naloxone (concentration at 15 min = 1599 nM) had extremely high concentrations at early post-administration time points. Although OIRD was reversed in both treatment groups after the first fentanyl dose, rats treated with naloxone exhibited significant hyperventilation and experienced severe convulsions / seizures. No hyperventilation or seizures were observed in animals treated with AG10-L-E2-naloxone. Pharmacokinetic data showed that both free naloxone and naloxone released from AG10-L-E2-naloxone had significantly higher plasma concentrations at 6 hr (750 nM and 60 nM, respectively) and 24 hr (395 nM and 44 nM, respectively). Figure 4f However, although AG10-L-E2-naloxone showed mild activity after the second fentanyl dose, both treatment groups demonstrated effective protection against OIRD after the second (6 hr) and third (24 hr) doses of fentanyl. Figures 4g to 4i This data suggests that exposing rats to very high doses of naloxone following fentanyl administration can actually cause some initial harm or alterations in μ / other receptors, thus reducing the animals' protection from naloxone at later time points. There are reports of pulmonary edema following high / rapid reversal. However, further investigation is needed.

[0516] Example 8: AG10-L1-E2-naloxone as a potential preventative agent against fentanyl-induced death and OIRD. To compensate for the lower efficacy of AG10-L-E2-naloxone in the first 30 minutes after administration, we tested the feasibility of combining the prodrug with a small amount of free naloxone. AG10-L-E2-naloxone (60 µmol kg -1 Equivalent to 63 mg kg -1 ) and naloxone (3 µmol kg -1 Equivalent to 1 mg kg -1 A single subcutaneous dose combination yielded excellent efficacy data, with OIRD completely reversed after all three fentanyl doses. Figures 5a to 5d Importantly, no rats experienced severe hyperventilation or convulsions / seizures, indicating that AG10-L-E2-naloxone poses a low risk in producing opioid-induced withdrawal symptoms.

[0517] Finally, we explored the potential of using AG10-L-E2-naloxone as a prophylactic agent against fentanyl. Figures 5e to 5g First, rats (each treatment group) were given... n = 6) Administer AG10-L-E2-naloxone (60 µmol kg -1 ) or naloxone (15µmol kg) -1 and 60 µmol kg -1 Minute ventilation remained unchanged compared to animals treated with the medium. After 6 hours, a single dose of fentanyl (1.5 mg / kg) was administered. -1 Rats were attacked with a subcutaneous dose. AG10-L-E2-naloxone was the only antidote that significantly protected rats from OIRD, and all animals (…) n = 6 Survival. In contrast, neither of the two naloxone doses was effective in preventing OIR...

Claims

1. A conjugate of formula (I): Or its pharmaceutically acceptable salt, wherein: T is a ligand selective for transthyretin-TTR; L stands for connector; L B Click products that do not exist or are formed through a click reaction between the first click handle and the second click handle; E is a cleavable ester; and D 1 This refers to the pharmaceutical component.

2. The conjugate as claimed in claim 1, wherein: L B The following are the click products: (i) Copper-catalyzed reactions between azides and alkynes; (ii) The reaction between azides and dibenzocyclooctene (DBCO); (iii) Reverse electron-demanding Diels-Alder cycloaddition (IEDDA) between the anti-cyclooctene (TCO) moiety and the tetrazine ring, or (iv) Staudinger reaction between azides and phosphine.

3. The conjugate of claim 1, wherein the conjugate comprises the following portion: , , or , Each R 7 Independently selected from the following groups: hydrogen, halogen atoms (e.g., Cl, Br, or F), hydroxyl group, -OCF3, -NH2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 Cycloalkyl, p is 0, 1, 2 or 3, and Indicates the connection point with the connector (L).

4. The conjugate of claim 1, wherein T has the structure of compound (A): Or its pharmaceutically acceptable salt, wherein: X a X b and X c Independently selected from C(R) 8a (R) 8b ), O, NR 8b Or S; where R 8a and R 8b Independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted aminoacyl, amino, substituted amino, amide, substituted amide and cyano; The A ring is a 4- to 12-membered ring. In some embodiments, the 4- to 12-membered ring is an aromatic ring or a heteroaromatic ring. Each Y is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted amino, amino, substituted amino, amide, substituted amide, sulfonamide, sulfonyl fluoride, thioester and cyano; c is an integer in the range of 0 to 5; and, Ring B is a heterocycle selected from the following (h1-h30): Where R 11 To R 16 Independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted aminoacyl, amino, substituted amino, amide, substituted amide, and cyano; R 17 Selected from hydroxyl, alkyl, amino, and alkylamino; and R 11 To R 16 At least one of them is related to X c Linking groups.

5. The conjugate of claim 1, wherein T has the structure of compound (B): Or its pharmaceutically acceptable salt, wherein: n is an integer in the range of 0 to 8; R 18 R 19 and R 20 Independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted amino, amino, substituted amino, amide, substituted amide, sulfonamide, sulfonyl fluoride, thioester and cyano; X a For C(R) 21 (R) 22 ), O, NR 22 Or S; where R 21 and R 22 Independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted aminoacyl, amino, substituted amino, amide, substituted amide, and cyano; A is a 5- to 12-membered ring; in some embodiments, the 5- to 12-membered ring is an aromatic ring or a heteroaromatic ring. Each Y is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted amino, amino, substituted amino, amide, substituted amide, sulfonamide, sulfonyl fluoride, thioester, and cyano; and c is a number from 0 to 5.

6. The conjugate of claim 1, wherein T has the structure of compound (C): (C)， Or its pharmaceutically acceptable salt, wherein: n is an integer in the range of 1 to 4; R 23 It is a short-chain alkyl group having 1 to 4 carbon atoms; R 24 It is hydrogen; R 25 It is a short-chain alkyl group having 1 to 4 carbon atoms; X a For C(R) 21 (R) 22 ), O, NR 22 Or S; where R 21 and R 22 Independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted aminoacyl, amino, substituted amino, amide, substituted amide, and cyano; Each Y is independently selected from hydrogen, halogen, acyl, substituted acyl, carboxyl, heterocyclic, alkoxycarbonyl, sulfonamide, sulfonyl fluoride, thioester, and substituted alkoxycarbonyl; and c is 2.

7. The conjugate of claim 1, wherein T has the structure of compound (D): (D)， Or its pharmaceutically acceptable salt, wherein: n is between 1 and 8; R 23 R 24 and R 25 Independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted aminoacyl, amino, substituted amino, amide, substituted amide, and cyano; X a For C(R) 21 (R) 22 ), O, NR 22 Or S; where R 21 and R 22 Independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted aminoacyl, amino, substituted amino, amide, substituted amide, and cyano; R a For CHO, COOH, COOCH3, COOR 6 CONR 7 R 8 Tetrazolyl, CONHOH, B(OH)2, CONHSO2Ar, CONHCH(R) 9 COOH, CF3, hydrogen, halogen, alkyl, substituted alkyl, acyl, substituted acyl, carboxyl, heterocyclic, sulfonamide, sulfonyl fluoride, thioester, alkoxycarbonyl or substituted alkoxycarbonyl; R b For CHO, COOH, COOCH3, COOR2 6 CONR2 7 R2 8 Tetrazolyl, CONHOH, B(OH)2, CONHSO2Ar, CONHCH(R2) 9 COOH, CF3, hydrogen, halogen, alkyl, substituted alkyl, acyl, substituted acyl, carboxyl, heterocyclic, sulfonamide, sulfonyl fluoride, thioester, alkoxycarbonyl or substituted alkoxycarbonyl; R2 6 It is an alkyl, haloalkyl, cycloalkyl, or heterocyclic group; R 27 and R 28 Each is independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, or heteroaryl; and, R 29 It is a side chain of a naturally occurring α-aminocarboxylic acid.

8. The conjugate of claim 7, wherein T has the structure of compound (E): (E) Or its pharmaceutically acceptable salt.

9. The conjugate of claim 7, wherein T has the structure of compound (Y): (Y) Or its pharmaceutically acceptable salt.

10. A conjugate of formula (II'): (II'), Or its pharmaceutically acceptable salt, wherein: Each R 1 Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n is 0, 1, 2, 3 or 4; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4 alkyl; L stands for connector; Y 1 For key or C 1-4 Alkylene; Y 2 For NR 6 , O or S; R 6 For H or C 1-4 alkyl; Each R 7 Choose independently from the following groups: hydrogen, halogen atoms (e.g., Cl, Br, or F), NO2, CF3, OMe, O(C 1-4 Alkyl), hydroxyl, -OCF3, -NH2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 cycloalkyl; p is 0, 1, 2, or 3; and D 1 This refers to the pharmaceutical component.

11. A (III') conjugate: (III') Or its pharmaceutically acceptable salt, wherein: Each R 1 Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n is 0, 1, 2, 3 or 4; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4 alkyl; L A C 3-20 Alkylene, wherein The C 3-20 Alkylene is optionally surrounded by one or more L 1a Replace, and One or more alkylene units are optionally selected from -O-, -NR y Substitution of - and -S- groups; Each L 1a Halogenated, C 1-6 Alkyl groups, or C groups substituted with halogen groups 1-6 alkyl; Each R y For H or C 1-4 alkyl; L B The click product is formed by the click reaction between the first click handle and the second click handle; Y 1 For key or C 1-4 Alkylene; Y 2 For NR 6 , O or S; R 6 For H or C 1-4 alkyl; Each R 7 Choose independently from the following groups: hydrogen, halogen atoms (e.g., Cl, Br, or F), NO2, CF3, OMe, O(C 1-4 Alkyl), hydroxyl, -OCF3, -NH2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 cycloalkyl; p is 0, 1, 2, or 3; and D 1 This refers to the pharmaceutical component.

12. A (IV') conjugate: (IV') Or its pharmaceutically acceptable salt, wherein: Each R 1 Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n is 0, 1, 2, 3 or 4; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4 alkyl; L A C 3-20 Alkylene, wherein The C 3-20 Alkylene is optionally surrounded by one or more L 1a Replace, and One or more alkylene units are optionally selected from -O-, -NR y Substitution of - and -S- groups; Each L 1a Halogenated, C 1-6 Alkyl groups, or C groups substituted with halogen groups 1-6 alkyl; Each R y For H or C 1-4 alkyl; L B The click product is formed by the click reaction between the first click handle and the second click handle; Y 1 For key or C 1-4 Alkylene; Y 2 For NR 6 , O or S; R 6 For H or C 1-4 alkyl; Each R 7 Choose independently from the following groups: hydrogen, halogen atoms (e.g., Cl, Br, or F), NO2, CF3, OMe, O(C 1-4 Alkyl), hydroxyl, -OCF3, -NH2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 Cycloalkyl; p is 0, 1, 2 or 3; and D 1 This refers to the pharmaceutical component.

13. A (V') conjugate: (V')， Or its pharmaceutically acceptable salt, wherein: Each R 1 Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n is 0, 1, 2, or 3; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4 alkyl; L A C 3-20 Alkylene, wherein The C 3-20 Alkylene is optionally surrounded by one or more L 1a Replace, and One or more alkylene units are optionally selected from -O-, -NR y Substitution of - and -S- groups; Each L 1a Halogenated, C 1-6 Alkyl groups, or C groups substituted with halogen groups 1-6 alkyl; Each R y For H or C 1-4 alkyl; Y 1 For key or C 1-4 Alkylene; Y 2 For NR 6 , O or S; R 6 For H or C 1-4 alkyl; Each R 7 Independently selected from the following groups: hydrogen, halogen atoms (e.g., Cl, Br, or F), hydroxyl group, -OCF3, -NH2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 Cycloalkyl; p is 0, 1, 2 or 3; and D 1 This refers to the pharmaceutical component.

14. A (VI') conjugate: (WE') Or its pharmaceutically acceptable salt, wherein: Each R 1 Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n is 0, 1, 2, or 3; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4 alkyl; L A C 3-20 Alkylene, wherein The C 3-20 Alkylene is optionally surrounded by one or more L 1a Replace, and One or more alkylene units are optionally selected from -O-, -NR y Substitution of - and -S- groups; Each L 1a Halogenated, C 1-6 Alkyl groups, or C groups substituted with halogen groups 1-6 alkyl; Each R y For H or C 1-4 alkyl; Y 1 For key or C 1-4 Alkylene; Y 2 For NR 6 , O or S; R 6 For H or C 1-4 alkyl; Each R 7 Independently selected from the following groups: hydrogen, halogen atoms (e.g., Cl, Br, or F), hydroxyl group, -OCF3, -NH2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 cycloalkyl; p is 0, 1, 2, or 3; and D 1 The drug component (opioid agonist or opioid antagonist).

15. A conjugate of formula (VII'): (VII'), Or its pharmaceutically acceptable salt, wherein: Each R 1 Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n is 0, 1, 2, or 3; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4 alkyl; Y 1 For key or C 1-4 Alkylene; Y 2 For NR 6 , O or S; L B The click product is formed by the click reaction between the first click handle and the second click handle; R 6 For H or C 1-4 alkyl; Each R 7 Independently selected from the following groups: hydrogen, halogen atoms (e.g., Cl, Br, or F), hydroxyl group, -OCF3, -NH2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 cycloalkyl; p is 0, 1, 2 or 3; r is 1, 2, 3, 4, 5, 6, or 7; s is 1, 2, 3, 4, 5, 6, or 7; and D 1 This refers to the pharmaceutical component.

16. A conjugate of formula (VIII'): (VIII') Or its pharmaceutically acceptable salt, wherein: Each R 1 Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n is 0, 1, 2, or 3; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4 alkyl; Y 1 For key or C 1-4 Alkylene; Y 2 For NR 6 , O or S; L B The click product is formed by the click reaction between the first click handle and the second click handle; R 6 For H or C 1-4 alkyl; Each R 7 Independently selected from the following groups: hydrogen, halogen atoms (e.g., Cl, Br, or F), hydroxyl group, -OCF3, -NH2, -NHC 1-4 Alkyl, -N(C) 1-4 Alkyl group 2, -CONH2, -CONHC 1-4 Alkyl, -CON(C) 1-4 Alkyl)2、-NHCOC 1-4 Alkyl, C 1-6 Alkyl, C 1-6 Alkoxy and C 3-6 cycloalkyl; p is 0, 1, 2 or 3; r is 1, 2, 3, 4, 5, 6, or 7; s can be 1, 2, 3, 4, 5, 6, or 7; t is 1, 2, 3, or 4; u is 1, 2, 3, or 4; and D 1 This refers to the pharmaceutical component.

17. A conjugate of formula (IX): (IX), Or its pharmaceutically acceptable salt, wherein: Each R 1 Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n is 0, 1, 2, or 3; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4 alkyl; L stands for connector; Z 1 C 1-6 Alkyl, C 6-10 Aryl, or 5 to 10-membered heteroaryl, among which The C 1-6 Alkyl groups are optionally surrounded by one or more Z 1a Replace, and The C 6-10 Aryl or 5- to 10-membered heteroaryl groups are optionally surrounded by one or more Z groups. 1b replace; Each Z 1a or Z 1b Independently halogenated, C 1-6 Alkoxy, or C, optionally substituted with a halogen group. 1-6 Alkyl; and D 1 The drug component (opioid agonist or opioid antagonist).

18. A conjugate of formula (X): (X)， Or its pharmaceutically acceptable salt, wherein: Each R 1 Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n is 0, 1, 2, or 3; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4 alkyl; L A C 3-20 Alkylene, wherein The C 3-20 Alkylene is optionally surrounded by one or more L 1a Replace, and One or more alkylene units are optionally selected from -O-, -NR y Substitution of - and -S- groups; Each L 1a Halogenated, C 1-6 Alkyl groups, or C groups substituted with halogen groups 1-6 alkyl; Each R y For H or C 1-4 alkyl; L B The click product is formed by the click reaction between the first click handle and the second click handle; Z 1 C 1-6 Alkyl, C 6-10 Aryl, or 5 to 10-membered heteroaryl, among which The C 1-6 Alkyl groups are optionally surrounded by one or more Z 1a Replace, and The C 6-10 Aryl or 5- to 10-membered heteroaryl groups are optionally surrounded by one or more Z groups. 1b replace; Each Z 1a or Z 1b Independently halogenated, C 1-6 Alkoxy, or C, optionally substituted with a halogen group. 1-6 Alkyl; and D 1 This refers to the pharmaceutical component.

19. A conjugate of formula (XI): (XI), Or its pharmaceutically acceptable salt, wherein: Each R 1 Independently a halogenated or C 1-4 alkyl; m can be 0, 1, 2, or 3; R 2 For H or C 1-4 alkyl; X 1 -O- or -NR x -; n is 0, 1, 2, or 3; R 3 For H or C 1-4 alkyl; R 4 For H or C 1-4 alkyl; R 5 For H or C 1-4 alkyl; R x For H or C 1-4 alkyl; L A C 3-20 Alkylene, wherein The C 3-20 Alkylene is optionally surrounded by one or more L 1a Replace, and One or more alkylene units are optionally selected from -O-, -NR y Substitution of - and -S- groups; Each L 1a Halogenated, C 1-6 Alkyl groups, or C groups substituted with halogen groups 1-6 alkyl; Each R y For H or C 1-4 alkyl; Z 1 C 1-6 Alkyl, C 6-10 Aryl, or 5 to 10-membered heteroaryl, among which The C 1-6 Alkyl groups are optionally surrounded by one or more Z 1a Replace, and The C 6-10 Aryl or 5- to 10-membered heteroaryl groups are optionally surrounded by one or more Z groups. 1b replace; Each Z 1a or Z 1b Independently halogenated, C 1-6 Alkoxy, or C, optionally substituted with a halogen group. 1-6 Alkyl; and D 1 This refers to the pharmaceutical component.

20. The conjugate according to any one of claims 1-19, wherein D 1 It is a peptide.

21. The conjugate according to any one of claims 1-19, wherein D 1 It can be a small molecule, nucleotide, oligonucleotide, polynucleotide, amino acid, oligopeptide, polypeptide or protein.

22. The conjugate according to any one of claims 1-19, wherein D 1 It is a cytokine.

23. The conjugate of claim 22, wherein the cytokine is interleukin-2 (IL-2).

24. The conjugate of claim 22, wherein the cytokine is interleukin-15 (IL-15).

25. The conjugate according to any one of claims 1-19, wherein D 1 It is an anticancer agent.

26. The conjugate of claim 25, wherein the anticancer agent is SN-38, irinotecan, capecitabine, monomethylolpropionate E, trifluuridine, docetaxel, or infinitib.

27. The conjugate according to any one of claims 1-19, wherein D 1 It is insulin.

28. The conjugate according to any one of claims 1-19, wherein D 1 It is a glucagon-like peptide-1 (GLP-1) agonist.

29. The conjugate of claim 28, wherein the GLP-1 agonist is smegglutide, liraglutide, duraglutide, abiglutide, or exenatide.

30. The conjugate according to any one of claims 1-19, wherein D 1 It is an agonist of glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic peptide (GIP) receptors.

31. The conjugate of claim 30, wherein glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic peptide (GIP) receptor agonist are telpoide.

32. A method of treating a disease, the method comprising applying the conjugate according to any one of claims 1-31.

33. The method of claim 32, further comprising administering a second drug (D) in a free (unconjugated) form. 2 ).

34. The method of claim 33, wherein D 1 With D 2 same.

35. The method of claim 33, wherein D 1 With D 2 different.

36. A method of treating cancer, the method comprising applying the conjugate of claim 25 or claim 26.

37. The method of claim 36, further comprising administering a second drug (D) in a free (unconjugated) form. 2 ).

38. The method of claim 37, wherein D 1 With D 2 same.

39. The method of claim 37, wherein D 1 With D 2 different.

40. A method for treating obesity, the method comprising administering the conjugate according to any one of claims 28-31.

41. A method for treating diabetes, the method comprising administering the conjugate according to any one of claims 28-31.

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