Phenylmaleimide linkers

Abstract

Compounds are disclosed that are useful as linker-payload compounds. The compounds have the following structure (I) as stereoisomers, enantiomers or tautomers thereof or mixtures thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein X1, X2, X3, X4, X5, R4a and R4b are as defined herein. Additional compounds, conjugates, methods of preparation, pharmaceutical compositions, and methods of treatment related to conjugates of a compound of structure (I) and a targeting moiety or binding fragment thereof are also provided. # imgabs0 #

Description

Background Art

[0001] Antibody-drug conjugates (ADCs) are a class of interesting bioconjugates. ADCs used for the treatment of certain cancers combine the targeting characteristics of monoclonal antibodies with the cancer killing ability of cytotoxic drugs to provide several advantages over other non-targeted chemotherapeutic agents. However, challenges associated with the complexity of ADC molecules, particularly the linker between the targeting peptide and the cytotoxic drug, have caused significant complexity to the progress and development of new and effective therapeutic agents. Although the first ADC was approved in 2001, it took nearly a decade for the next ADC to be approved. The next generation of ADC molecules now contain non-cytotoxic payloads, such as immuno-oncology drugs, to stimulate the innate and adaptive immune systems, thereby attacking tumors from the inside.

[0002] There is a need for chemical linkers that can enhance the effectiveness of targeted therapeutic molecules (e.g., ADCs). The present disclosure satisfies this need and provides additional related advantages. BRIEF DESCRIPTION OF THE DRAWINGS

[0003] In the drawings, the same reference numerals identify similar elements (e.g., the conjugates of Table 2). The sizes and relative positions of the elements in the drawings are not necessarily drawn to scale. For example, the shapes of the various elements and angles are not drawn to scale, and some of the elements are enlarged and positioned to improve the readability of the drawings. In addition, the particular shapes of the elements depicted are not intended to convey any information about the actual shapes of the particular elements, and they are selected only for ease of identification in the drawings.

[0004] Figure 1A It was shown that the conjugate II-12 induced TNF-α production by human PBMCs in the presence of the HER2-expressing SK-BR-3 tumor cell line in a dose-dependent manner.

[0005] Figure 1B It was shown that conjugate II-12 induced insufficient TNF-α production in human PBMCs in the presence of the MDA-MB-48 cell line, which does not express HER2.

[0006] Figure 2A It was shown that conjugates II-15, II-1, II-16 and II-17 induced TNF-α production by human PBMCs in a dose-dependent manner in the presence of the MDA-MB-175-VII tumor cell line expressing fibronectin-4.

[0007] Figure 2BIt is shown that in the presence of HEK-293 cells lacking fibronectin-4 expression, the production of TNF-α in human PBMCs induced by conjugates II-15, II-1, II-16 and II-17 is deficient.

[0008] Figure 3A It was shown that the conjugates II-18, II-19, II-19, II-20, II-21 induced TNF-α production by human PBMCs in a dose-dependent manner in the presence of the MDA-MB-175-VII tumor cell line expressing fibronectin-4.

[0009] Figure 3B It is shown that the TNF-α production in human PBMCs induced by the conjugates II-18, II-19, II-19, II-20, II-21 is deficient in the presence of HEK-293 cells lacking fibronectin-4 expression.

[0010] Figure 4A It was shown that conjugate II-3 induced TNF-α production by human PBMCs in the presence of the MDA-MB-175-VII tumor cell line expressing fibronectin-4 in a dose-dependent manner.

[0011] Figure 4B It is shown that in the presence of HEK-293 cells lacking fibronectin-4 expression, the production of TNF-α in human PBMCs induced by conjugate II-3 is deficient.

[0012] Figure 5A It was shown that conjugate II-2 induced TNF-α production by human PBMCs in a dose-dependent manner in the presence of the MDA-MB-175-VII tumor cell line expressing fibronectin-4.

[0013] Figure 5B It is shown that in the presence of HEK-293 cells lacking fibronectin-4 expression, the TNF-α production in human PBMCs induced by conjugate II-2 is deficient. DETAILED DESCRIPTION

[0014] The present disclosure provides a branched phenylmaleimide compound (e.g., a compound of structure (I)), to facilitate loading of a bioactive molecule (also referred to herein as a payload) onto a targeting compound, such as an antibody or a ligand. The present disclosure further provides the use of a linker compound for preparing a linker-payload to be conjugated with a targeting moiety (e.g., an antibody, a fusion protein, a ligand). Compared with being connected in series (or linearly) to a payload, the advantage of using a hydrophilic part (e.g., polar, hydrophilic, charged functional group, etc.) to bind to a phenylmaleimide by being parallel to the payload topology is that stability is increased when used as part of a conjugate, and off-target uptake or activity is minimized by hiding or covering the payload hydrophobicity. In addition, when the conjugate is present in the systemic circulation, such a structure seems to maximize the possibility that the drug-antibody ratio (DAR) will generally remain fixed (that is, the conjugate stability will be maintained while maintaining favorable pharmacokinetic properties). For example, when the thiol present in cysteine ​​is conjugated to the compound of structure (I), a succinimide substituted by a thioether is formed. While not wishing to be bound by theory, it is believed that due to the above-mentioned substituted phenyl groups of structure (I), this succinimide hydrolyzes rapidly, and this hydrolyzed succinimide is no longer able to deconjugate via a retro-Michael reaction. Therefore, the linker-payload is "locked" to the protein and tends to better retain the DAR.

[0015] Before describing the present disclosure in more detail, it may be helpful to provide definitions of certain terms used herein. Additional definitions are set forth throughout the present disclosure.

[0016] As used in the specification and claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. It should be understood that, as used herein, the terms "a" and "an" refer to "one or more" of the listed components.

[0017] The use of alternatives (eg, "or") should be understood to mean one, two, or any combination of the alternatives.

[0018] As used herein in the context of a number, the term "about" refers to a range centered on the number and spanning less than 15% of the number and greater than 15% of the number. The term "about" used in the context of a range refers to an extended range that is 15% less than the lowest value listed in the range and 15% greater than the maximum value listed in the range. In some embodiments, a given value refers to a range of values ​​(i.e., "about" a given value). For example, in some embodiments, pH 7.4 refers to approximately pH 7.4 (i.e., a range of 6.3 to 8.5).

[0019] Throughout the disclosure, unless otherwise stated, any concentration range, percentage range, ratio range, or integer range should be understood to include the value of any integer within the range, and where appropriate, fractions thereof (e.g., tenths and hundredths of integers). Furthermore, unless otherwise stated, any numerical range of the disclosure relating to any physical characteristic such as polymer subunits, size, or thickness should be understood to include any integer within the stated range. Throughout the disclosure, numerical ranges are inclusive of their cited endpoints unless otherwise specifically stated.

[0020] Unless the context requires otherwise, throughout this specification and claims, the word "comprise" and variations thereof (such as "comprises" and "comprising") are to be interpreted in an open, inclusive sense, i.e., to mean "including, but not limited to." As used herein, the terms "comprise" and "comprising" are used synonymously.

[0021] The phrase "at least one of" when followed by a list of items or elements refers to an open-ended set of one or more elements in the list, which may, but does not necessarily, contain more than one element.

[0022] As used herein, "variant" proteins or polypeptides include one or more non-natural amino acids, one or more amino acid substitutions, one or more amino acid insertions, one or more amino acid deletions, or any combination thereof, which may occur at one or more sites relative to a reference polypeptide disclosed herein, and wherein the variant protein or polypeptide has substantially similar activity (e.g., enzymatic function, immunogenicity) relative to the reference polypeptide. As determined by the sequence alignment programs and parameters disclosed herein, the variant proteins or polypeptides disclosed herein have at least about 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher sequence identity to the amino acid sequence of the reference polypeptide disclosed herein. Variant polypeptides may be produced, for example, by genetic polymorphism or artificial manipulation. Conservative substitutions of amino acids are well known and may occur naturally or may be engineered when a protein is produced recombinantly. Amino acid substitutions, deletions and additions can be introduced into proteins using mutagenesis methods known in the art (see, e.g., Sambrook et al., Molecular Cloning: A Laboratory Manual, 3rd ed., Cold Spring Harbor Laboratory Press, NY, 2001). Oligonucleotide-directed site-specific (or fragment-specific) mutagenesis procedures can be used to generate polynucleotides with altered codons that provide the desired substitutions, deletions or insertions. Alternatively, random or saturation mutagenesis techniques, such as alanine scanning mutagenesis, error-prone polymerase chain reaction mutagenesis or oligonucleotide-directed mutagenesis, can be used to prepare polypeptide variants (see, e.g., Sambrook et al., supra).

[0023] In some embodiments, a "binding domain" or "binding region" or "targeting moiety" refers to a protein, polypeptide, oligopeptide, peptide, carbohydrate, nucleic acid, or combination thereof that is capable of specifically binding to one or more targets (e.g., adhesion protein-4, MSLN, HER2, LRRC15, ASGR1, CD40 or BAFF and / or APRIL). A binding domain comprises any naturally occurring, synthetic, semisynthetic, or recombinantly produced binding partner of a biological molecule of interest or another target. Exemplary binding domains of the present disclosure include, for example, Fab', F(ab'), 2 , Fab, Fv, rIgG, scFv, hcAb (heavy chain antibody), single domain antibody, V HH 、V NAR, sdAb, nanobodies, receptor extracellular domains or their ligand binding portions or ligands (e.g., cytokines, chemokines). "Fab" (antigen binding fragment) is the part of an antibody that binds to an antigen and comprises the variable region and CH1 of a heavy chain connected to a light chain by an interchain disulfide bond. Various assays for identifying binding domains of the present disclosure that specifically bind to a particular target are known, including Western blots, ELISAs, and Analysis). Particularly preferred binding domains include immunoglobulin light and heavy chain variable domains (e.g., scFv, Fab), and are referred to herein as "immunoglobulin binding domains" or "immunoglobulin binding proteins". Immunoglobulin binding domains can be incorporated into a variety of protein scaffolds or structures as described herein (such as antibodies or antigen-binding fragments thereof, scFv-Fc fusion proteins, or fusion proteins comprising two or more such immunoglobulin binding domains).

[0024] Throughout the disclosure, the term "antibody" refers to an immunoglobulin molecule that specifically binds to an antigen or is immunoreactive to an antigen. The portion of an antibody that binds to an antigen may be referred to as an "antigen binding domain". In certain embodiments, an antibody is a complete antibody, including at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds, and an antigen-binding portion of a complete antibody that has or retains the ability to bind to a target molecule. The antibody or antigen-binding fragment thereof disclosed herein may include, for example, a polyclonal antibody, a monoclonal antibody, and a genetically engineered antibody. The monoclonal antibody or antigen-binding portion thereof disclosed herein may be non-human (e.g., mouse, rabbit), chimeric, humanized, or human. In certain embodiments, the antibody disclosed herein is a heterologous conjugate antibody, a bispecific antibody, a multispecific antibody, a double antibody, a tri-antibody, or a tetra-antibody. Immunoglobulin structure and function are reviewed in, for example, Greenfield, ed., Antibodies: A Laboratory Manual, Chapters 2 and 3 (Cold Spring Harbor Laboratory, Cold Spring Harbor, 2014).

[0025] For example, the term "V L ” and “V H ” refer to the variable binding regions of the antibody light chain and heavy chain, respectively. The variable binding region is composed of discrete, well-defined subregions called “complementarity determining region” (CDR) and “framework region” (FR). The term “CL” refers to “immunoglobulin light chain constant region” or “light chain constant region”, i.e., the constant region from the antibody light chain. The term “CH” refers to “immunoglobulin heavy chain constant region” or “heavy chain constant region”, which can be further divided into CH1, CH2 and CH3 (IgA, IgD, IgG) or CH1, CH2, CH3 and CH4 domains (IgE, IgM), depending on the antibody isotype.

[0026] If the binding domain and its fusion protein are equal to or greater than 10 5 M -1 Affinity or K a (i.e., the equilibrium binding constant for a specific binding interaction in units of 1 / M) to a target without significantly binding to other components present in the test sample, then it "specifically binds" to a target. Binding domains (or their fusion proteins) can be classified as "high affinity" binding domains (or their fusion proteins) and "low affinity" binding domains (or their fusion proteins). A "high affinity" binding domain refers to a K a For at least 10 8 M -1 , at least 10 9 M -1 , at least 10 10 M -1 , at least 10 11 M -1 , at least 10 12 M -1 or at least 10 13 M -1 , preferably at least 10 8 M -1 or at least 10 9 M -1 "Low affinity" binding domains are those with a K a For up to 10 8 M -1 , up to 10 7 M -1 , up to 10 6 M -1 , up to 10 5 M -1 Alternatively, affinity can be defined as the equilibrium dissociation constant (K d ), in M ​​(for example, 10 -5 M to 10 -13 M). The affinity of binding domain polypeptides and fusion proteins according to the present disclosure can be readily determined using conventional techniques (see, e.g., Scatchard et al., Ann. NY Acad. Sci. 51:660, 1949; and U.S. Pat. Nos. 5,283,173, 5,468,614, or equivalents).

[0027] As used herein, "derivative" refers to a chemically or biologically modified form of a compound that is structurally similar to a parent compound and (actually or theoretically) can be derived from the parent compound. Generally, the difference between a "derivative" and an "analog" is that a parent compound can be the starting material for producing a "derivative", while the parent compound is not necessarily the starting material for producing an "analog". An analog may have different chemical or physical properties from the parent compound. For example, a derivative may be more hydrophilic than the parent compound, or it may have altered reactivity (e.g., a CDR with an amino acid change that alters its affinity for a target).

[0028] As used herein, "same" or "identity" refers to the similarity between a DNA, RNA, nucleotide, amino acid or protein sequence and another DNA, RNA, nucleotide, amino acid or protein sequence. Identity can be expressed as the percentage of the sequence identity of the first sequence and the second sequence. The percentage of sequence identity (%) relative to the reference DNA sequence can be the percentage of DNA nucleotides in the candidate sequence that are identical to the DNA nucleotides in the reference DNA sequence after sequence alignment. The percentage of sequence identity (%) relative to the reference amino acid sequence can be the percentage of amino acid residues in the candidate sequence that are identical to the amino acid residues in the reference amino acid sequence after aligning the sequences and introducing gaps (if necessary) to achieve the maximum percentage of sequence identity and without considering any conservative substitutions as part of the sequence identity. For example, percent sequence identity values ​​for the sequences provided herein can be generated using NCBIBLAST 2.0 software as defined in Altschul et al., "Gapped BLAST and PSI-BLAST: a new generation of protein database search programs", Nucleic Acids Res. 2007, 25, 3389-3402, with the parameters set to the default values.

[0029] "Oxo" refers to a radical of formula =0.

[0030] "Nitro" refers to the molecular formula -NO 2 The group.

[0031] "Thio" refers to a radical of formula =S.

[0032] "Cyano" refers to a group of formula -CN.

[0033] "Amino" refers to the molecular formula -NH 2The group.

[0034] "Hydroxyl" or "hydroxyl" refers to a radical of the molecular formula -OH.

[0035] "Thiol" refers to a group of the molecular formula -SH.

[0036] "Aldehyde" refers to a radical of the molecular formula -C(=O)H.

[0037] "Carboxyl" refers to a functional group of the molecular formula -C(=O)OH.

[0038] "Halo" refers to a halogen group, such as -F, -Cl, -Br, -I, and the like.

[0039] "Alkyl" refers to a saturated, straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, having from one to twelve carbon atoms (C 1 -C 12 Alkyl), one to eight carbon atoms (C 1 -C 8 Alkyl) or one to six carbon atoms (C 1 -C 6 alkyl) or any value within these ranges, such as C 4 -C 6 Alkyl, alkyl, etc., and it is connected to the rest of the molecule by a single bond (e.g., methyl, ethyl, n-propyl, 1 methylethyl (isopropyl), n-butyl, n-pentyl, 1,1 dimethylethyl (tert-butyl), 3 methylhexyl, 2 methylhexyl, etc.). The carbon number referred to relates to the carbon backbone and carbon branches, but does not include carbon atoms belonging to any substituent. Similarly, alkenyl refers to a straight or branched hydrocarbon chain group consisting of carbon and hydrogen with at least one carbon-carbon double bond. "Alkynyl" contains at least one carbon-carbon triple bond. Unless otherwise specifically stated in the specification, alkyl, alkenyl or alkynyl are optionally substituted.

[0040] "Haloalkyl" refers to an alkyl group as defined above substituted with one or more halo groups as defined above, for example, trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl, etc. Unless stated otherwise specifically in the specification, a haloalkyl group is optionally substituted.

[0041] "Alkoxy" refers to a group of the formula -OR a A group in which R a is an alkyl group as defined above containing from one to twelve carbon atoms (C 1 -C 12 Alkoxy), one to eight carbon atoms (C 1 -C 8Alkoxy) or one to six carbon atoms (C 1 -C 6 Unless stated otherwise specifically in the specification, an alkoxy group is optionally substituted.

[0042] "Haloalkoxy" refers to an alkoxy group as defined above substituted with one or more halo groups as defined above. Unless stated otherwise specifically in the specification, a haloalkoxy group is optionally substituted.

[0043] "Aminyl" refers to a group of the formula -NR a R b A group in which R a and R b are each independently H or C as defined above 1 -C 6 Alkyl. When R a and R b When both are H, "amino" is the same as "amino" as defined above. Unless otherwise specified, the C 1 -C 6 The alkyl moiety is optionally substituted.

[0044] "Amindyl" refers to a group of the formula -C(=O)NR a R b A group in which R a and R b are each independently H or C as defined above 1 -C 6 Alternatively, amide may also refer to a group or -N(R a )C(=O)R b , where R a Is H or C 1 -C 6 Alkyl, and R b It is C 1 -C 6 Unless otherwise specified, the C 1 -C 6 The alkyl moiety is optionally substituted.

[0045] "Carbohydrate" refers to a group consisting of carbon atoms, hydrogen atoms, and oxygen atoms. In some embodiments, the carbohydrate has a hydrogen to oxygen ratio of 2:1. In some embodiments, the carbohydrate has an empirical formula of C m (H 2 O) n, wherein m and n are integers that may be the same or different. Exemplary carbohydrates include sugars (e.g., monosaccharides, disaccharides, oligosaccharides, polysaccharides), starch, and cellulose. In some embodiments, the carbohydrate is selected from the group consisting of glucose, fructose, sucrose, ribose, straight-chain starch, lactose, galactose, xylose, maltose, isomaltulose, trehalose, sorbitol, mannitol, maltodextrin, raffinose, stachyose, straight-chain starch, amylopectin, glycogen, cellulose, hemicellulose, pectin, hydrocolloids, etc.

[0046] "Haloalkyl" refers to an alkyl group as defined above that is substituted with one or more halo groups. The haloalkyl group is attached to the backbone via an alkyl carbon atom. Unless specifically stated otherwise in the specification, a haloalkyl group is optionally substituted.

[0047] "Carboxyalkyl" refers to an alkyl group as defined above substituted by one or more carboxyl groups. The carboxyalkyl group is attached to the backbone via an alkyl carbon atom. Unless otherwise specifically stated in the specification, the carboxyalkyl group is optionally substituted.

[0048] "Cycloalkyl" or "carbocycle" refers to a stable non-aromatic monocyclic or polycyclic hydrocarbon radical consisting only of carbon and hydrogen atoms, which may contain a fused or bridged ring system, having three to fifteen carbon atoms, preferably three to ten carbon atoms, and which is saturated or unsaturated and is connected to the rest of the molecule by a single bond. Monocyclic cycloalkyls include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic groups include, for example, adamantyl, norbornyl, decahydronaphthyl, 7,7-dimethyl-bicyclo[2.2.1]heptyl, etc. Unless otherwise specifically stated in the specification, cycloalkyls may be optionally substituted.

[0049] "Aryl" refers to any stable monocyclic, bicyclic or polycyclic carbocyclic ring system having from 4 to 12 atoms in each ring, wherein at least one ring is aromatic. Some examples of aryl include phenyl (sometimes represented as "Ph"), naphthyl, tetrahydronaphthyl, indanyl, anthracenyl and biphenyl. In the case where the aryl substituent is bicyclic and one ring is non-aromatic, it is understood that attachment is through the aromatic ring. Aryl can be substituted or unsubstituted.

[0050] The term "heterocycle" or "heterocyclyl" refers to an aromatic or non-aromatic ring system of five to twenty-two atoms, wherein 1 to 4 ring atoms are heteroatoms selected from oxygen, nitrogen and sulfur. Thus, the heterocycle can be a heteroaryl or its dihydro or tetrahydro form. Heterocycles include pyrrolidine, tetrahydrofuran, thiolane, indolinyl, 3H-indolyl, azetidine, oxetane, thietane, diazetidine, dioxetane, dithietane, piperidine, tetrahydrofuran, pyran, tetrahydropyran, thiocyclohexane, tetrahydrothiophene, pyridine, pyrimidine, etc.

[0051] "Heteroaryl" refers to any stable monocyclic, bicyclic or polycyclic carbocyclic ring system having 4 to 12 atoms in each ring, wherein at least one ring is aromatic and contains 1 to 4 heteroatoms selected from oxygen, nitrogen and sulfur. Some examples of heteroaryl include acridinyl, quinoxalinyl, pyrazolyl, indolyl, benzotriazolyl, furanyl, thienyl, benzothienyl, benzofuranyl, quinolyl, isoquinolyl, oxazolyl, isoxazolyl, pyrazinyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl and tetrahydroquinolyl. Heteroaryl includes N-oxide derivatives of nitrogen-containing heteroaryl.

[0052] "Alkylene" or "alkylene chain" refers to a straight or branched divalent hydrocarbon chain connecting the rest of the molecule to a group consisting of only carbon and hydrogen, containing no unsaturation and having from one to twelve carbon atoms, for example, methylene, ethylene, propylene, n-butene, etc. The alkylene chain is connected to the rest of the molecule by a single bond and to the group by a single bond. The points of attachment of the alkylene chain to the rest of the molecule and to the group can be through one carbon or any two carbons within the chain. Unless otherwise specifically stated in the specification, alkylene is optionally substituted.

[0053] Similarly, "alkenylene" refers to a straight or branched divalent hydrocarbon chain that connects the rest of the molecule to a radical, consists solely of carbon and hydrogen, and contains at least one carbon-carbon double bond. The alkenylene chain may contain from two to twelve carbon atoms. "Alkynylenyl" also refers to a straight or branched divalent hydrocarbon chain that connects the rest of the molecule to a radical, consists solely of carbon and hydrogen, but contains at least one carbon-carbon triple bond.

[0054] "Heteroalkylene" refers to an alkylene group as defined above, including at least one heteroatom (e.g., Si, N, O, P, or S) within the alkylene chain or at the terminus of the alkylene chain. In some embodiments, the heteroatom is within the alkylene chain (i.e., the heteroalkylene group includes at least one carbon-[heteroatom] x-carbon bond, wherein x is 1, 2 or 3). In other embodiments, the heteroatom is located at the end of the alkylene group, and is therefore used to join the alkylene group to the remainder of the molecule (e.g., M1-HA-M2, wherein M1 and M2 are part of the molecule, H is a heteroatom, and A is an alkylene group). Unless otherwise specifically stated in the specification, sub-assorted alkylene is optionally substituted. Exemplary sub-assorted alkylenes include ethylene oxide (e.g., polyethylene oxide), propylene oxide, amino acid chains (i.e., peptides short to medium length containing 1-15 amino acids) and alkylene chains connected by various functional groups, such as amides, disulfides, phosphates, sulfates, sulfonamides, esters, ethers, -S-, carbamates, ureas, thioureas, anhydrides, etc. (including combinations thereof). In some embodiments, sub-assorted alkylenes include polyamino acids with 1-10 amino acids. In some embodiments, sub-assorted alkylenes include polyamino acids with 1-5 amino acids.

[0055] "Heteroalkenylene" refers to a heteroalkylene group as defined above containing at least one carbon-carbon double bond. "Heteroalkynylene" refers to a heteroalkylene group as defined above containing at least one carbon-carbon triple bond.

[0056] "Heteroatom linker" refers to a continuous heteroatom chain or heteroatom that connects a portion of a molecule to a group. A heteroatom linker can be multivalent (e.g., divalent, trivalent, etc.). A heteroatom linker consists only of non-carbon atoms (e.g., H, O, N, S, Si, P, etc.). Unless otherwise specifically stated in the specification, a heteroatom linker can be optionally substituted.

[0057] "Cycloalkylene" is a polyvalent (eg, divalent, trivalent, etc.) cycloalkyl group. Unless specifically stated otherwise in the specification, a cycloalkylene group may be optionally substituted.

[0058] "Arylene" is a polyvalent (eg, divalent, trivalent, etc.) aromatic group. Unless specifically stated otherwise in the specification, an arylene group may be optionally substituted.

[0059] "Heterocyclylene" is a polyvalent (eg, divalent, trivalent, etc.) heterocyclyl group. Unless stated otherwise specifically in the specification, a heterocyclylene group may be optionally substituted.

[0060] "Heteroarylene" is a polyvalent (eg, divalent, trivalent, etc.) heteroarylene group. Unless specifically stated otherwise in the specification, a heteroarylene group may be optionally substituted.

[0061] "Joint" refers to a continuous chain of at least one atom (such as carbon, oxygen, silicon, nitrogen, sulfur, phosphorus and combinations thereof), which connects a part of a molecule to another part of the same molecule or different molecules or fragments thereof by a covalent bond (e.g., a single bond, a double bond or a triple bond). In some embodiments, the joint is an optionally substituted alkylene joint, an optionally substituted alkenylene joint, an optionally substituted alkynylene joint, an optionally substituted sub-assorted alkyl joint, an optionally substituted sub-assorted alkenyl joint, an optionally substituted sub-assorted alkynyl joint, a heteroatom joint, a cycloalkylene joint, an arylene joint, a sub-heterocyclyl joint, a sub-heteroaryl joint or a combination thereof. Unless otherwise specifically stated in the specification, the joint may be optionally substituted.

[0062] "Trigger element" refers to a molecular motif (e.g., acid-labile) that is recognized by a biomolecule (e.g., a protease, such as a cathepsin) or is sensitive to a chemical reaction in a biological environment. Typically, enzymatic recognition of a trigger element produces a catalytic cleavage reaction that releases the payload from the antibody+linker.

[0063] "Sacrificial element" or "self-sacrificial spacer" refers to a chemically unstable group that promotes the release or cleavage between the parts of the molecule (e.g., between the antibody-linker and the payload). The sacrificial element is typically a bivalent linker motif that is subject to one or more chemical reactions, ultimately resulting in the cleavage of the covalent bond between the two ends of the group. The reaction occurs due to a certain type of stimulation of the system (e.g., when the antibody-drug conjugate is subjected to the intracellular environment, the protease catalyzes the reaction during its recognition of the trigger element or pH reduction), which then leads to the controlled release of the payload. Commonly used sacrificial elements are p-aminobenzyloxycarbonyl (PABC) and aminal. In some embodiments, the sacrificial element has a structure in the following structures:

[0064]

[0065]

[0066] The following scheme depicts the fragmentation of p-aminobenzyloxycarbonyl (PABC) and release of the payload:

[0067]

[0068] Where D represents the unmodified payload.

[0069] In some embodiments, the sacrificial element has the following structure:

[0070]

[0071] in:

[0072] R 6a , R6b , R 6c and R 6d is independently hydrogen, optionally substituted alkyl, optionally substituted aryl or optionally substituted heteroaryl, or

[0073] R 6a and R 6c together with the nitrogen and carbon atoms to which they are attached form an azetidinyl, pyrrolidinyl, piperidinyl or homopiperidinyl group, and R 6d is hydrogen; and

[0074] Y 1 Is –O–, –S–, or –NR 6b –.

[0075] In some embodiments, the sacrificial element has the following structure:

[0076]

[0077] in:

[0078] R 6e , R 6f , R 6g and R 6h is independently hydrogen, optionally substituted alkyl, optionally substituted aryl or optionally substituted heteroaryl, or

[0079] R 6a and R 6c together with the nitrogen and carbon atoms to which they are attached form an azetidinyl, pyrrolidinyl, piperidinyl or homopiperidinyl group, and R 6d is hydrogen; and

[0080] Y 2 Is –O–, –S–, or –NR 6f –.

[0081] In certain embodiments, the sacrificial element has the following structure:

[0082]

[0083] in:

[0084] R 10 is independently at each occurrence alkyl, alkoxy or halo;

[0085] R 11 is hydrogen, alkyl or –(CH 2 CH 2 O) z3 -CH 3 ;

[0086] R 12 is hydrogen or alkyl;

[0087] R 13 is hydrogen or alkyl;

[0088] z1 is 0 or 1;

[0089] z2 is 0, 1, 2, 3 or 4; and

[0090] z3 is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0091] In some embodiments, the sacrificial element has one of the following structures:

[0092]

[0093] in:

[0094] R 14a , R 14b , R 14c , R 14d , R 14e and R 14f are each independently hydrogen, alkyl, hydroxyalkyl or alkoxyalkyl;

[0095] z4, z5, z6 and z7 are each independently 1, 2, 3, 4, 5 or 6.

[0096] The aforementioned terms include all systems that include effective cleavage elements in the endolysosomal processing of antibody-drug conjugates. Exemplary systems include enzymatic peptide sequences based on cathepsins, such as Val-Cit, Val-Ala, Ala-Ala, Gly-Gly-Phe-Gly, etc. Also included are trigger elements sensitive to b-glucuronidase, which contain glucuronic acid as a trigger element. In some embodiments, the trigger element is directly and covalently bound to the sacrificial element. In some embodiments, the sacrificial element is directly and covalently bound to the payload, and once cleavage occurs, the payload is irreversibly released.

[0097] A "non-cleavable element" refers to a linker that contains neither a trigger element nor a sacrificial element and is not cleaved under normal physiological conditions. Depending on the target, many conjugates are very effective without a formal cleavage release element. Therefore, in some embodiments, the payload can be directly linked to the phenylmaleimide via a non-cleavable linker.

[0098] As defined above, "sub-heteroalkyl element" refers to a joint comprising one or more sub-heteroalkyl groups. In some embodiments, sub-heteroalkyl elements can be used to optimize the properties of joint-payload. In some embodiments, sub-heteroalkyl elements increase the linear connection between other elements (e.g., charged elements, hydrophilic elements, etc.). In some embodiments, sub-heteroalkyl elements increase the distance between the polar cap or the payload of the phenyl-maleimide part of structure (I). Sub-heteroalkyl elements vary in length, structure, polarity, degree of branching, etc. The above variables allow joint-payload and then antibody-joint-drug conjugates to have optimal overall properties (e.g., solubility, monodispersity, etc.).

[0099] In some embodiments, the heteroalkylene element includes a PEG having two ethylene glycol units (ie, PEG 2 ) and a linear polyethylene glycol of one or more amino acids. In another embodiment, the heteroalkylene element comprises a Gly-Gly dipeptide. In some embodiments, the heteroalkylene element comprises PEG 1-24 , dipeptides, tripeptides and tetrapeptides or combinations thereof. In other embodiments, the heteroalkylene element comprises an amino acid. In another embodiment, the heteroalkylene element comprises a PEG. 1-24 In another embodiment, the heteroalkylene element comprises PEG 1-12 In another embodiment, the heteroalkylene element comprises PEG 2-6 In some embodiments, PEG x The two are combined with amino acids to form heteroalkylene elements. Exemplary linear PEG moieties can be found, for example, in PCT Publication No. WO 2021 / 207701, which is hereby incorporated by reference in its entirety.

[0100] "Hydrophilic element" refers to a part of structure (I) that effectively promotes the solubility of the entire molecule in an aqueous solvent (e.g., water, phosphate buffered saline). In fact, the hydrophilic element hides or offsets the hydrophobic part of the linker-payload, making the resulting molecule stable in an aqueous environment (e.g., pH 7.4 buffered water). In some embodiments, the hydrophilic element produces a stable and soluble linker-payload with improved overall physicochemical properties. The hydrophilic element provides a suitable chemical function that enables stable conjugation with a protein. In some embodiments, incorporation of an appropriate hydrophilic element makes the resulting protein conjugate have a more protein-like ADME / DMPK distribution.

[0101] In some embodiments, the hydrophilic element comprises a medium length PEG linker (10-14 units or PEG 10-14 In some embodiments, the hydrophilic element comprises 2-24 units of PEG (ie, PEG2-24 In some embodiments, the length and number of branches included in the hydrophilic element can be adjusted based on the other elements present in structure (I). In some embodiments, the hydrophilic element is C 1 -C 6 Alkoxy (e.g., methoxy). In some embodiments, the hydrophilic element comprises polysarcosine (PSAR). On the one hand, the hydrophilic element imparts polarity and hydrophilicity to the entire conjugate. In some embodiments, the hydrophilic element is large enough in size and flexible enough in structure to mask the hydrophobicity with or without a trigger element. In some embodiments, the hydrophilic element is divalent. In some embodiments, the hydrophilic element is a monovalent group. In some embodiments, the hydrophilic element is multivalent. In some embodiments, the hydrophilic element is branched. In some embodiments, the hydrophilic element is linear.

[0102] Hydrophilic groups (e.g., forming part of a hydrophilic element) are known to those of ordinary skill in the art. For example, hydrophilic groups can be found in PCT Publication Nos. WO 2019 / 217591, WO 2018 / 089373, and the hydrophilic groups from each disclosure are hereby incorporated by reference in their entirety.

[0103] A "polar cap" refers to a structure having one or more polar or hydrophilic functional groups (e.g., one or more -OH, -NH 2 、-C(=O)OH、-S(O) 3 、-OP(O) 3In some embodiments, the polar cap is connected to the end of structure (I). In some embodiments, the polar cap is connected to more than one end of structure (I) (for example, if the polar cap is connected to multiple ends of a branched heteroalkylene element or a branched hydrophilic element). The polar cap imparts hydrophilicity to the entire compound of structure (I). In some embodiments, the polar cap includes one or more parts that are negatively charged under physiological conditions (for example, phosphates, carboxylates, sulfates, sulfonic acids, etc.). In some embodiments, the polar cap includes one or more parts that are positively charged under physiological conditions (for example, quaternary amines). In some embodiments, under physiological conditions, the polar cap itself is zwitterionic, with an overall negative charge, an overall positive charge, or an overall neutral charge. In some embodiments, the polar cap is used to increase both polarity and charge by incorporating amino acids, which may have additional functions to increase further polarity and charge. In some embodiments, the polar cap comprises L-glutamic acid. In some embodiments, the polar cap has two carboxyl groups. In some embodiments, the polar cap comprises one or more diacids. In some embodiments, the glutamic acid is further modified such that one or both acids are amidated with a polyol-containing amine. In some embodiments, the polar cap includes an amino acid with a glycosidic moiety (eg, L-serine-β-D-glucoside, etc.).

[0104] "Stable compound" and "stable structure" are meant to indicate a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture and formulation into an efficacious therapeutic agent.

[0105] "Salt" includes both acid addition salts (e.g., salts formed with inorganic or organic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, acetic acid, ascorbic acid, etc.) and base addition salts (e.g., salts formed with inorganic or organic bases such as sodium, potassium, amine bases, etc.).

[0106] Crystallization can produce solvates of the compounds described herein (e.g., compounds of structure (I)). Embodiments of the present disclosure include all solvates of the described compounds. As used herein, the term "solvate" refers to an aggregate comprising one or more molecules of a compound of the present disclosure and one or more molecules of a solvent.

[0107] Embodiments of the compounds of the present disclosure (e.g., compounds of structure (I)) or salts, tautomers or solvates thereof may contain one or more stereocenters and may therefore give rise to enantiomers, diastereomers and other stereoisomeric forms, which may be defined in terms of absolute stereochemistry as (R)- or (S)- or (D)- or (L)- of an amino acid. Embodiments of the present disclosure are intended to encompass all such possible isomers, as well as racemic and optically pure forms thereof. Optically active (+) and (-), (R)- and (S)- or (D)- and (L)-isomers may be prepared using chiral synthons or chiral reagents or resolved using conventional techniques, e.g., chromatography and fractional crystallization. Conventional techniques for preparing / isolating individual enantiomers include chiral synthesis from suitable optically pure precursors or resolution of the racemate (or racemate of a salt or derivative) using, for example, chiral high pressure liquid chromatography (HPLC). When the compounds described herein contain olefinic double bonds or other features that result in geometric asymmetry, and unless specified otherwise, it is intended that the compounds include both E and Z geometric isomers. Likewise, all tautomeric forms are intended to be included.

[0108] "Stereoisomers" refer to compounds composed of the same atoms joined by the same bonds but with different three-dimensional structures, which are not interchangeable. The present disclosure contemplates various stereoisomers and mixtures thereof and includes "enantiomers," which refer to two stereoisomers whose molecules are non-superimposable mirror images of each other.

[0109] "Tautomer" refers to a proton transfer from one atom of a molecule to another atom of the same molecule. The present disclosure includes tautomers of any of the compounds. Various tautomeric forms of the compounds can be easily derived by one of ordinary skill in the art.

[0110] The chemical nomenclature schemes and structure diagrams used herein are a modified form of the IUPAC nomenclature system using the ACD / Name Version 9.07 software program and / or the ChemDraw Ultra Version 11.0 software nomenclature program (Cambridge Software). Common names familiar to those of ordinary skill in the art are also used.

[0111] For ease of illustration, various compounds of structure (I) or conjugates of structure (II) with sulfur- or phosphorus-containing moieties (e.g., sulfonates, phosphates, etc.) can be present in an anionic state (e.g., -S(O) 3 or –OP(O) 3Those skilled in the art will readily appreciate that the charge is dependent on pH, and that uncharged forms (e.g., protonated forms or salt forms, such as sodium or other cations) are also included within the scope of the embodiments of the present disclosure.

[0112] Joint Compounds

[0113] In certain embodiments, the disclosure provides a branched phenylmaleimide compound (e.g., a compound of structure (I)) that is capable of forming a covalent bond between a linker-payload and a protein. As described above, in certain embodiments of the disclosure, a compound that can be used as a linker between a payload (including a fragment thereof) and a targeting peptide (including a fragment thereof, e.g., an antibody) is provided.

[0114] Therefore, some embodiments provide a compound having the following structure (I):

[0115]

[0116] in:

[0117] X 1 , X 2 , X 3 , X 4 and X 5 One of them is CL 1 -R 1 , X 1 , X 2 , X 3 , X 4 and X 5 The other one is CL 2 -R 2 , and X 1 , X 2 , X 3 , X 4 and X 5 The other three are N, CR, 3 or CL 3 -R 3a ;

[0118] R 1 , R 2 and R 3a Each independently comprises one or more moieties selected from the group consisting of amino acid elements, charged elements, heteroalkylene elements, hydrophilic elements, trigger elements, sacrificial elements, polar caps, payloads, and combinations thereof;

[0119] The condition is R 1 and R 2 At least one of includes a payload;

[0120] R 3 is independently selected at each occurrence from the group consisting of hydrogen, deuterium, alkyl, haloalkyl, halo, alkoxy, haloalkoxy, amino, amine, amido, aldehyde, hydroxyl, cyano, nitro, thiol, carboxyl, carboxyalkyl, alkyl-S(O) 3 H, Alkyl-OP(O) 3 H, Alkyl-P(O) 3 H, -O-carboxyalkyl, -O-alkyl-S(O) 3 H, -O-alkyl-OP(O) 3 H, -O-alkyl-P(O) 3 H, -S(O) 3 H, -OP(O) 3 H, -P(O) 3 H, Alkyl-OP(O) 3 -Alkyl, Alkyl-P(O) 3 -alkyl, -O-alkyl-S(O) 3 -alkyl, -O-alkyl-OP(O) 3 -alkyl, -O-alkyl-P(O) 3 -alkyl, -S(O) 3 -alkyl, -OP(O) 3 -Alkyl, -P(O) 3 -Alkyl groups, sulfamides, sulfenamides and carbohydrates;

[0121] R 4a and R 4b are independently hydrogen, deuterium, halogenated or -SR 4c , where R 4c is substituted or unsubstituted C 6 -C 10 aryl or substituted or unsubstituted 5-12 membered heteroaryl; and

[0122] L 1 , L 2 and L 3 each independently is a linker comprising an optionally substituted alkylene, an optionally substituted alkenylene, an optionally substituted alkynylene, an optionally substituted heteroalkylene, an optionally substituted heteroalkenylene, an optionally substituted heteroalkynylene, a heteroatom linker, an optionally substituted cycloalkylene, an optionally substituted arylene, an optionally substituted heterocyclylene, an optionally substituted heteroarylene, or a combination thereof;

[0123] As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0124] Certain embodiments provide a compound having the following structure (I):

[0125]

[0126] in:

[0127] X 1 , X 2 , X 3 , X 4 and X 5 One of them is CL 1 -R 1 , X 1 , X 2 , X 3 , X 4 and X 5 The other one is CL 2 -R 2 , and X 1 , X 2 , X 3 , X 4 and X 5 The other three are N, CR, 3 or CL 3 -R 3a ;

[0128] R 1 , R 2 and R 3a Each independently comprises one or more moieties selected from the group consisting of amino acid elements, charged elements, heteroalkylene elements, hydrophilic elements, trigger elements, sacrificial elements, polar caps, payloads, and combinations thereof;

[0129] The condition is R 1 and R 2 At least one of includes a payload;

[0130] R 3 is independently selected at each occurrence from the group consisting of hydrogen, deuterium, alkyl, haloalkyl, halo, alkoxy, haloalkoxy, amino, amine, amido, aldehyde, hydroxyl, cyano, nitro, thiol, carboxyl, carboxyalkyl, alkyl-S(O) 3 H, Alkyl-OP(O) 3 H, Alkyl-P(O) 3 H, -O-carboxyalkyl, -O-alkyl-S(O) 3 H, -O-alkyl-OP(O) 3 H, -O-alkyl-P(O) 3 H, -S(O) 3 H, -OP(O) 3 H, -P(O) 3 H, Alkyl-OP(O)3 -Alkyl, Alkyl-P(O) 3 -alkyl, -O-alkyl-S(O) 3 -alkyl, -O-alkyl-OP(O) 3 -alkyl, -O-alkyl-P(O) 3 -alkyl, -S(O) 3 -alkyl, -OP(O) 3 -Alkyl, -P(O) 3 -Alkyl groups, sulfamides, sulfenamides and carbohydrates;

[0131] R 4a and R 4b are independently hydrogen, deuterium, halogenated or -SR 4c , where R 4c is substituted or unsubstituted C 6 -C 10 aryl or substituted or unsubstituted 5-12 membered heteroaryl; and

[0132] L 1 , L 2 and L 3 each independently is a direct bond or a linker comprising an optionally substituted alkylene, an optionally substituted alkenylene, an optionally substituted alkynylene, an optionally substituted heteroalkylene, an optionally substituted heteroalkenylene, an optionally substituted heteroalkynylene, a heteroatom linker, an optionally substituted cycloalkylene, an optionally substituted arylene, an optionally substituted heterocyclylene, an optionally substituted heteroarylene, or a combination thereof;

[0133] As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0134] In some embodiments, R 4a and R 4b In some embodiments, R 4a is halogenated, or R 4b In some embodiments, R 4a , R 4b or both have one of the following structures:

[0135]

[0136] In some embodiments, R 1 , R 2 and / or R 3aThe invention comprises an element selected from the group consisting of an amino acid element, a charged element, an alkylene element, a hydrophilic element, a trigger element, a sacrificial element, a polar cap, a payload, and combinations thereof. It should be understood that these elements can be connected in any order and in a linear manner or through branched connections. In some embodiments, R 1 , R 2 and / or R 3a Multiple occurrences of an element are included (eg, two or more heteroalkylene elements, two or more hydrophilic elements, two or more polar caps, etc.).

[0137] In some embodiments, R 1 , R 2 or R 3a A branch point is included as part of an amino acid element (e.g., lysine), wherein additional elements are attached via the epsilon amine of the lysine, and other additional elements are attached to the amino acid element via one or more peptide bonds at the alpha carbon of the lysine. A similar motif can be used with glutamic acid of the amino acid element. In some embodiments, the amino acid element includes one of the following structures:

[0138]

[0139] In some embodiments, the compound of structure (I) comprises one of the following structures:

[0140]

[0141]

[0142] In certain embodiments, R 1 Has the following structure:

[0143]

[0144] in:

[0145] L 1a It is an amino acid element;

[0146] L 1b It is a live component;

[0147] L 1c is a heteroalkylene element;

[0148] L 1d It is a hydrophilic element;

[0149] L 1e is a triggering element; and

[0150] L 1f It is a sacrificial component;

[0151] Where L 1a , L 1b , L 1c , L 1d , L 1e and L 1f When present in one or more instances, optionally with L 1a , L 1b , L 1c , L 1d , L 1e and L 1f One or more of the other of the are joined to form one or more rings;

[0152] n1, n2, n3, n4, n5 and n6 are independently an integer from 0 to 3 at each occurrence, provided that n1+n2+n3+n4+n5+n6=1 or greater;

[0153] n7 is 1, 2, 3, 4, 5 or 6; and

[0154] R 1a Yes and L 1a , L 1b , L 1c , L 1d , L 1e or L 1f A payload is covalently bound in one occurrence and is optionally replaced by a polar cap.

[0155] In some embodiments, n7 is 1, 2, or 3. In some embodiments, n7 is 1 or 2. In some embodiments, n7 is 1.

[0156] In some embodiments, R 1 Has the following structure:

[0157]

[0158] in:

[0159] L 1a It is an amino acid element;

[0160] L 1b It is a live component;

[0161] L 1c is a heteroalkylene element;

[0162] L 1d It is a hydrophilic element;

[0163] L 1e is a triggering element; and

[0164] L 1f It is a sacrificial component;

[0165] Where L 1a , L 1b , L 1c , L 1d , L 1e and L 1f When present in one or more instances, optionally with L 1a , L 1b , L 1c , L 1d , L 1e and L 1f One or more of the other of the are joined to form one or more rings;

[0166] n1, n2, n3, n4, n5 and n6 are independently an integer from 0 to 3 at each occurrence, provided that n1+n2+n3+n4+n5+n6=1 or greater;

[0167] n7 is 1, 2, 3, 4, 5 or 6; and

[0168] R 1a is the payload that is optionally replaced by the polar cap.

[0169] In further embodiments, R 2 Has the following structure:

[0170]

[0171] in:

[0172] L 2a It is an amino acid element;

[0173] L 2b It is a live component;

[0174] L 2c is a heteroalkylene element;

[0175] L 2d It is a hydrophilic element;

[0176] L 2e It is the trigger element;

[0177] m1, m2, m3, m4 and m5 are independently an integer from 0 to 3 at each occurrence, provided that m1+m2+m3+m4+m5=1 or greater;

[0178] m6 is 1, 2, 3, 4, or 5; and

[0179] R 2a is hydrogen, alkyl, payload, or polar cap.

[0180] In some embodiments, m6 is 1, 2, or 3. In some embodiments, m6 is 1 or 2. In some embodiments, m6 is 1.

[0181] In still more embodiments, R 3a Has the following structure:

[0182]

[0183] in:

[0184] L 3a It is an amino acid element;

[0185] L 3b It is a live component;

[0186] L 3c is a heteroalkylene element;

[0187] L 3d It is a hydrophilic element;

[0188] L 3e It is the trigger element;

[0189] p1, p2, p3, p4 and p5 are independently an integer from 0 to 3 at each occurrence, provided that p1+p2+p3+p4+p5=1 or greater;

[0190] p6 is 1, 2, 3, 4, or 5; and

[0191] R 3b is hydrogen, alkyl or polar cap.

[0192] In some embodiments, p6 is 1, 2, or 3. In some embodiments, p6 is 1 or 2. In some embodiments, p6 is 1.

[0193] In certain embodiments, n7 is 1, and each of n1 to n6 is 1. In certain embodiments, n7 is 1, and each of n1 to n4 is 0, n5 is 1, and n6 is 1. In certain embodiments, n7 is 1, and n1 is 0, n2 is 0, n3 is 1, n4 is 0, n5 is 1, and n6 is 1. In certain embodiments, n7 is 1, and n1 is 1, n2 is 0, n3 is 1, n4 is 0, n5 is 1, and n6 is 1. In certain embodiments, n7 is 1, and n1 is 1, n2 is 1, n3 is 1, n4 is 0, n5 is 1, and n6 is 1. In certain embodiments, n7 is 1, and n1 is 1, n2 is 1, n3 is 1, n4 is 0, n5 is 1, and n6 is 1. In certain embodiments, n7 is 1, and n1 is 1, n2 is 0, n3 is 1, n4 is 0, n5 is 1, and n6 is 1. In certain embodiments, n7 is 2. In certain embodiments, n7 is 3.

[0194] In some embodiments, m6 is 1, and each of m1 to m5 is 1. In some embodiments, m6 is 1, m1 is 1, m2 is 0, m3 is 0, m4 is 1, and m5 is 0. In some embodiments, m6 is 1, m1 is 1, m2 is 1, m3 is 0, m4 is 1, and m5 is 0. In some embodiments, m6 is 1, m1 is 1, m2 is 0, m3 is 1, m4 is 1, and m5 is 0. In some embodiments, m6 is 1, m1 is 1, m2 is 0, m3 is 1, m4 is 1, and m5 is 0. In some embodiments, m6 is 1, m1 is 1, m2 is 0, m3 is 0, m4 is 1, and m5 is 0. In some embodiments, m6 is 1, m1 is 1, m2 is 0, m3 is 0, m4 is 1, and m5 is 1. In some embodiments, m6 is 2. In certain embodiments, m6 is 3.

[0195] In some embodiments, p6 is 1, and each of p1 to p5 is 1. In certain embodiments, p6 is 1, p1 is 1, p2 is 0, p3 is 0, p4 is 1, and p5 is 0. In some embodiments, p6 is 1, p1 is 1, p2 is 1, p3 is 0, p4 is 1, and p5 is 0. In some embodiments, p6 is 1, p1 is 1, p2 is 0, p3 is 1, p4 is 1, and p5 is 0. In some embodiments, p6 is 1, p1 is 1, p2 is 0, p3 is 0, p4 is 1, and p5 is 0. In some embodiments, p6 is 1, p1 is 1, p2 is 0, p3 is 0, p4 is 1, and p5 is 1. In some embodiments, p6 is 2, and p1 is 1 at least once, p2 is 1, p3 is 0, p4 is 1, and p5 is 0. In some embodiments, p6 is 2. In certain embodiments, p6 is 3.

[0196] In some embodiments, the amino acid element includes one or more amino acids selected from the group consisting of glycine, alanine, serine, threonine, cysteine, valine, leucine, isoleucine, methionine, proline, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, asparagine, glutamine, histidine, lysine, arginine, sarcosine, and β-alanine.

[0197] In certain embodiments, the amino acid element is selected from the group consisting of glycine, sarcosine, β-alanine, and glutamic acid.

[0198] In some embodiments, the amino acid element comprises a dipeptide, a tripeptide, a tetrapeptide, or a pentapeptide.

[0199] In further embodiments, the amino acid element has one of the following structures:

[0200]

[0201] in:

[0202] R 5a is independently at each occurrence hydrogen, alkyl, hydroxyalkyl or alkoxyalkyl.

[0203] In some embodiments, the charged element includes a moiety having a negative charge at pH 7.4 (ie, a range of 6.3 to 8.5). In certain embodiments, the charged element includes a moiety having a positive charge at pH 7.4 (ie, a range of 6.3 to 8.5).

[0204] In some embodiments, the charged element includes one or more charged amino acids, one or more carboxylic acids, one or more sulfonic acids, one or more sulfonamides, one or more sulfates, one or more phosphates, one or more quaternary amines, one or more sulfamides, one or more sulfenamides, or combinations thereof.

[0205] In certain embodiments, the charged amino acid is aspartic acid, glutamic acid, histidine, lysine, or arginine.

[0206] In some embodiments, R 1 , R 2 or R 3a Non-cleavable linkers (eg, linkers or segments thereof that do not include a trigger element or a sacrificial element) are included.

[0207] In some embodiments, R 1 , R 2 or R 3a Includes one of the following structures:

[0208]

[0209] in:

[0210] R 5b , R 5c , R 5d and R 5e is independently selected at each occurrence from the group consisting of hydrogen, deuterium, alkyl, haloalkyl, halo, alkoxy, haloalkoxy, amino, hydroxy, cyano, nitro, thiol, carboxyalkyl, alkyl-S(O) 3 H, Alkyl-OP(O) 3 H, Alkyl-P(O) 3 H, -O-carboxyalkyl, -O-alkyl-S(O) 3 H, -O-alkyl-OP(O) 3 H, -O-alkyl-P(O) 3 H, -S(O) 3 H, -OP(O) 3 H, -P(O) 3 H, Alkyl-OP(O) 3 -Alkyl, Alkyl-P(O) 3 -alkyl, -O-alkyl-S(O) 3 -alkyl, -O-alkyl-OP(O) 3 -alkyl, -O-alkyl-P(O) 3 -alkyl, -S(O) 3 -alkyl, -OP(O) 3 -Alkyl, -P(O) 3 -Alkyl, sulfamide, sulfenamide;

[0211] R 5f is independently at each occurrence hydrogen, alkyl, hydroxyalkyl or alkoxyalkyl;

[0212] R 9 is independently at each occurrence hydrogen or alkyl;

[0213] q2 is independently an integer from 1 to 25 at each occurrence; and

[0214] q3 is independently an integer from 5 to 15 at each occurrence.

[0215] In some embodiments, the hydrophilic element comprises polyethylene glycol, polysarcosine, cyclodextrin, c-glycoside or a combination thereof. In some embodiments, the hydrophilic element comprises one of the following structures:

[0216]

[0217] in:

[0218] R 5b , R5c , R 5d and R 5e is independently selected at each occurrence from the group consisting of hydrogen, deuterium, alkyl, haloalkyl, halo, alkoxy, haloalkoxy, amino, hydroxy, cyano, nitro, thiol, carboxyalkyl, alkyl-S(O) 3 H, Alkyl-OP(O) 3 H, Alkyl-P(O) 3 H, -O-carboxyalkyl, -O-alkyl-S(O) 3 H, -O-alkyl-OP(O) 3 H, -O-alkyl-P(O) 3 H, -S(O) 3 H, -OP(O) 3 H and -P(O) 3 H,

[0219] R 5g is independently at each occurrence hydrogen, alkyl, hydroxyalkyl, or alkoxyalkyl; and

[0220] q4 is independently an integer from 1 to 24 at each occurrence.

[0221] In some embodiments, the hydrophilic element comprises one of the following structures:

[0222]

[0223] In some embodiments, the hydrophilic element comprises one of the following structures:

[0224]

[0225]

[0226] In some embodiments, the hydrophilic element has one of the following structures:

[0227]

[0228] In some embodiments, the hydrophilic element has the following structure:

[0229]

[0230] In some embodiments, the hydrophilic element has one of the following structures:

[0231]

[0232] In some embodiments, the hydrophilic element has one of the following structures:

[0233]

[0234]

[0235] In some embodiments, the hydrophilic element comprises polysarcosine. In some embodiments, the hydrophilic element is polysarcosine comprising the following structure:

[0236]

[0237] In some embodiments, the hydrophilic element is a polysarcosine having one of the following structures:

[0238]

[0239] In some embodiments, the molecular weight of the hydrophilic element is greater than 150 g / mole, greater than 200 g / mole, greater than 300 g / mole, greater than 400 g / mole, greater than 500 g / mole, greater than 600 g / mole, greater than 700 g / mole, greater than 800 g / mole, greater than 900 g / mole, or greater than 1000 g / mole. In some embodiments, the molecular weight of the hydrophilic element is less than 150 g / mole, less than 200 g / mole, less than 300 g / mole, less than 400 g / mole, less than 500 g / mole, less than 600 g / mole, less than 700 g / mole, less than 800 g / mole, less than 900 g / mole, or less than 1000 g / mole.

[0240] In certain embodiments, L 1 In some embodiments, L 1 It is C 1 -C 6 In certain embodiments, L 2 In some embodiments, L 2 It is C 1 -C 6 In certain embodiments, L 3 In some embodiments, L 3 It is C 1 -C 6 Alkylene.

[0241] In certain embodiments, L 1 In some embodiments, L 1 It is C 1 -C 6 In certain embodiments, L 2 In some embodiments, L 2 It is C 1-C 6 In certain embodiments, L 3 In some embodiments, L 3 It is C 1 -C 6 Heteroalkylene.

[0242] In further embodiments, L 1 , L 2 or L 3 It is C 1 -C 6 and N. In some embodiments, L 3 It is a direct key.

[0243] In further embodiments, L 1 , L 2 or L 3 Has one of the following structures:

[0244]

[0245] In some embodiments, the trigger element comprises a dipeptide, a tripeptide, a tetrapeptide, a pentapeptide, a glucuronide, a disulfide, a phosphate, a diphosphate, a triphosphate, a hydrazone, or a combination thereof. In some other embodiments, the trigger element comprises β-glucuronic acid. In certain embodiments, the trigger element comprises a dipeptide, a tripeptide, a tetrapeptide, or a pentapeptide. In some embodiments, the trigger element comprises two or more amino acids selected from the group consisting of: valine, citrulline, alanine, glycine, phenylalanine, lysine, or a combination thereof. In certain embodiments, the trigger element comprises a sequence of amino acids selected from the group consisting of: valine-citrulline, valine-alanine, glycine-glycine-phenylalanine-glycine, and a combination thereof. In some embodiments, the trigger element comprises a structure in the following structures, including a combination thereof:

[0246]

[0247]

[0248] In some embodiments, the molecular weight of the trigger element is greater than 150 g / mole, greater than 200 g / mole, greater than 300 g / mole, greater than 400 g / mole, greater than 500 g / mole, greater than 600 g / mole, greater than 700 g / mole, greater than 800 g / mole, greater than 900 g / mole, or greater than 1000 g / mole. In some embodiments, the molecular weight of the trigger element is less than 150 g / mole, less than 200 g / mole, less than 300 g / mole, less than 400 g / mole, less than 500 g / mole, less than 600 g / mole, less than 700 g / mole, less than 800 g / mole, less than 900 g / mole, or less than 1000 g / mole.

[0249] In some embodiments, the trigger element has the following structure:

[0250]

[0251] In some embodiments, the trigger element is specifically cut by an enzyme. For example, the trigger element can be cut by a lysosomal enzyme. The trigger element can be based on a peptide, or can include a peptide region that can serve as a substrate for the enzyme. Peptide-based trigger elements are more stable in plasma and extracellular environments than chemically unstable linkers.

[0252] An exemplary disulfide-containing trigger element may comprise the following structure:

[0253]

[0254] wherein D is the payload and R is independently selected at each occurrence from, for example, hydrogen or C 1 -C 6 Alkyl. Increasing the steric hindrance near the disulfide bond can increase the stability of the joint. When one or more R groups are selected from lower alkyl groups such as methyl, the above structure can increase the in vivo stability.

[0255] The peptide bond can have good serum stability because the activity of lysosomal proteolytic enzymes in the blood is very low due to endogenous inhibitors and unfavorable high pH values ​​of blood compared to lysosomes. Due to the effect of lysosomal proteases, such as cathepsin and plasmin, the payload can be released from the conjugate of structure (II). These proteases may exist at high levels in certain tumor tissues. The trigger element can be cut by lysosomal enzymes. Lysosomal enzymes can be, for example, cathepsin B, β-glucuronidase or β-galactosidase.

[0256] The cleavable peptide of the trigger element can be selected from a tetrapeptide such as Gly-Phe-Leu-Gly, Ala-Leu-Ala-Leu, a tripeptide such as Glu-Val-Cit or a dipeptide such as Val-Cit, Val-Ala, Ala-Ala and Phe-Lys. Compared with the longer peptide, the dipeptide can have a lower hydrophobicity.

[0257] In some embodiments, the trigger element can be a single amino acid residue. In some embodiments, the trigger element comprises Asn (eg, legumin cleavable).

[0258] The enzymatically cleavable trigger element can be combined with a sacrificial element to provide additional spatial separation between the payload and the enzymatic cleavage site. Direct connection of the payload to the peptide trigger element may result in proteolytic release of the payload or the amino acid adduct of the payload, thereby impairing its activity. The use of a sacrificial element can allow the release of a fully active, chemically unmodified payload upon amide bond hydrolysis.

[0259] The trigger element may contain chemically labile groups, such as hydrazone and / or disulfide groups. Trigger elements comprising chemically labile groups may exploit the differential properties between plasma and some cytoplasmic compartments. Intracellular conditions that may promote the release of payloads containing hydrazone trigger elements may be the acidic environment of endosomes and lysosomes, while disulfide-containing trigger elements may be reduced in the cytosol containing high thiol concentrations, such as glutathione. The plasma stability of trigger elements containing chemically labile groups may be improved by introducing steric hindrance using substituents near the chemically labile groups.

[0260] Acid labile groups, such as hydrazones, can remain intact during systemic circulation in the neutral pH environment of blood (pH 7.3-7.5), and can undergo hydrolysis, and once the conjugate of structure (II) is internalized into the weakly acidic endosomal (pH 5.0-6.5) and lysosomal (pH 4.5-5.0) compartments of the cell, the payload can be released. This pH-dependent release mechanism can be associated with non-specific release of the payload. In order to increase the stability of the hydrazone group of the trigger element, the trigger element can be altered by chemical modification, such as substitution, thereby allowing regulation to achieve more efficient release in the lysosome while minimizing losses in the circulation.

[0261] In some embodiments, the trigger element includes a hydrazone portion having one of the following structures:

[0262]

[0263] Where R is selected from C 1 -C 6 Alkyl, aryl and -OC1 -C 6 alkyl.

[0264] The hydrazone-containing trigger element may contain additional cleavage sites, such as additional acid-labile cleavage sites and / or enzyme-labile cleavage sites (e.g., disulfides). Conjugates and compounds including exemplary hydrazone-containing trigger elements may include, for example, the following structures:

[0265]

[0266] Where R is selected from C 1 -C 6 Alkyl, aryl and -OC 1 -C 6 alkyl.

[0267] Other acid labile groups that may be included in the trigger element include cis-aconityl containing linkers. Cis-aconityl chemistry can use a carboxylic acid juxtaposed to the amide bond to accelerate amide hydrolysis under acidic conditions.

[0268] The trigger element may also include a disulfide bond. Disulfide is thermodynamically stable at physiological pH, and releases a payload when the conjugate of structure (II) is internalized into the cell, wherein the cytosol can provide a significantly more reductive environment compared to the extracellular environment. The rupture of the disulfide bond may require the presence of a cytoplasmic sulfhydryl cofactor, such as (reduced) glutathione (GSH), so that the trigger element containing the disulfide bond is quite stable in the circulation, selectively releasing the payload in the cytosol. Intracellular enzyme protein disulfide isomerase or similar enzymes capable of cutting disulfide bonds may also contribute to the preferential cutting of intracellular disulfide bonds. GSH may be present in cells at a concentration range of 0.5-10mM, and by contrast, the concentration of about 5 μM of GSH or cysteine ​​(the most abundant low molecular weight thiol) in the circulation is significantly lower. Irregular blood flow may cause tumor cells in anoxic states to cause enhanced reductase activity, and therefore lead to even higher glutathione concentrations. The in vivo stability of disulfide bond-containing trigger elements can be enhanced by chemical modification of the trigger elements, for example, by exploiting steric hindrance near the disulfide bonds.

[0269] The trigger element can also be a β-glucuronic acid-based linker. The easy release of the payload can be achieved by the cleavage of the β-glucuronide glycosidic bond by the lysosomal enzyme β-glucuronidase. This enzyme can be present in large quantities in lysosomes and can be overexpressed in some tumor types, while the enzyme activity outside the cell may be low. Linkers based on β-glucuronic acid can be used to avoid the tendency of the conjugate to aggregate due to the hydrophilicity of β-glucuronide. In some embodiments, the trigger element includes β-glucuronic acid.

[0270] The following scheme depicts the release of payload (D) from a conjugate of structure (II) containing a β-glucuronic acid based trigger element:

[0271]

[0272] A variety of cleavable β-glucuronic acid-based linkers have been described that can be used to link drugs such as aurisin, camptothecin analogs, doxorubicin analogs, CBI minor groove binders, and psymberin to antibodies. Therefore, these β-glucuronic acid-based trigger elements are used in the conjugates of structure (II). In some embodiments, the trigger element includes a β-galactoside-based linker. β-galactoside is present in large quantities in lysosomes, while the enzyme activity outside the cell is very low.

[0273] The trigger element may comprise one or more peptides. In some embodiments, a peptide containing natural amino acids, unnatural amino acids, or any combination thereof may be selected. In some embodiments, the peptide may be a tripeptide or a dipeptide. In certain embodiments, the dipeptide comprises L-amino acids such as Val-Cit; Cit-Val; Ala-Ala; Ala-Cit; Cit-Ala; Asn-Cit; Cit-Asn; Cit-Cit; Val-Glu; Glu-Val; Ser-Cit; Cit-Ser; Lys-Cit; Cit-Lys; Asp-Cit; Cit-Asp; Ala-Val; Val-Ala; Phe-Lys; Lys-Phe; Val-Lys; Lys-Val; Ala-Lys; Lys-Ala; Phe-Cit; Cit-Phe; Leu-Cit; Cit-Leu; Ile-Cit; Cit-Ile; Phe-Arg; Arg-Phe; Cit-Trp; and Trp-Cit or a salt thereof.

[0274] Trigger elements and sacrificial groups are known in the art, for example in International Application No. PCT / US2021 / 054296; the trigger elements and sacrificial elements of the international application are hereby incorporated by reference in their entirety.

[0275] A sacrificial element can be a bifunctional p-aminobenzyl alcohol group, which can be linked to the trigger element via the amino group to form an amide bond, while an amine-containing payload can be linked to the benzyl hydroxyl group of p-aminobenzyl alcohol via a carbamate functional group (to give p-aminobenzyl carbamate). The resulting precursor compound can be activated under protease-mediated cleavage, resulting in a 1,6-elimination reaction, releasing the unmodified payload and the residue of the antibody-linker.

[0276] In some embodiments, the sacrificial element comprises p-aminobenzyloxycarbonyl, aminal, hydrazine, disulfide, amide, ester, hydrazine, phosphotriester, diester, β-glucuronide, double bond, triple bond, ether bond, ketone, diol, cyano, nitro, quaternary amine, or a combination thereof. In certain embodiments, the sacrificial element comprises p-methoxybenzyl, dialkyldialkoxysilane, diaryldialkoxysilane, orthoester, acetal, optionally substituted β-thiopropionate, ketal, phosphoramidate, hydrazone, vinyl ether, imine, aconityl, trityl, polyketal, bisarylhydrazone, diazobenzene, vicinal diol, pyrophosphate diester, or a combination thereof.

[0277] In certain embodiments, the sacrificial element comprises one of the following structures:

[0278]

[0279] In some embodiments, the sacrificial element comprises the following structure:

[0280]

[0281] in:

[0282] R 6a , R 6b , R 6c and R 6d is independently hydrogen, optionally substituted alkyl, optionally substituted aryl or optionally substituted heteroaryl, or

[0283] R 6a and R 6c together with the nitrogen and carbon atoms to which they are attached form an azetidinyl, pyrrolidinyl, piperidinyl or homopiperidinyl group, and R 6d is hydrogen; and

[0284] Y 1 Is –O–, –S–, or –NR 6b –

[0285] In certain embodiments, the sacrificial element comprises the following structure:

[0286]

[0287] in:

[0288] R 6e , R 6f , R 6g and R 6h is independently hydrogen, optionally substituted alkyl, optionally substituted aryl or optionally substituted heteroaryl, or

[0289] R 6a and R 6ctogether with the nitrogen and carbon atoms to which they are attached form an azetidinyl, pyrrolidinyl, piperidinyl or homopiperidinyl group, and R 6d is hydrogen; and

[0290] Y 2 Is –O–, –S–, or –NR 6f –

[0291] In some embodiments, the sacrificial element comprises the following structure:

[0292]

[0293] in:

[0294] R 10 is independently at each occurrence alkyl, alkoxy or halo;

[0295] R 11 is hydrogen, alkyl or –(CH 2 CH 2 O) z3 -CH 3 ;

[0296] R 12 is hydrogen or alkyl;

[0297] R 13 is hydrogen or alkyl;

[0298] z1 is 0 or 1;

[0299] z2 is 0, 1, 2, 3 or 4; and

[0300] z3 is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; or

[0301] In some embodiments, the sacrificial element comprises one of the following structures:

[0302]

[0303] in:

[0304] R 14a , R 14b , R 14c , R 14d , R 14e and R 14f are each independently hydrogen, alkyl, hydroxyalkyl or alkoxyalkyl;

[0305] z4, z5, z6 and z7 are each independently 1, 2, 3, 4, 5 or 6;

[0306] In some embodiments, the sacrificial element comprises one of the following structures:

[0307]

[0308] in:

[0309] z8 and z9 are each independently 1, 2, 3, 4, 5 or 6; or

[0310] In certain embodiments, the sacrificial element comprises one of the following structures:

[0311]

[0312] in:

[0313] R 15 is independently at each occurrence H, methyl, ethyl, isopropyl, tert-butyl or phenyl;

[0314] Y 3 is O or CH 2 ;and

[0315] q5 is an integer in the range of 1-5.

[0316] In some embodiments, the molecular weight of the sacrificial element is greater than 150 g / mole, greater than 200 g / mole, greater than 300 g / mole, greater than 400 g / mole, greater than 500 g / mole, greater than 600 g / mole, greater than 700 g / mole, greater than 800 g / mole, greater than 900 g / mole, or greater than 1000 g / mole. In some embodiments, the molecular weight of the sacrificial element is less than 150 g / mole, less than 200 g / mole, less than 300 g / mole, less than 400 g / mole, less than 500 g / mole, less than 600 g / mole, less than 700 g / mole, less than 800 g / mole, less than 900 g / mole, or less than 1000 g / mole.

[0317] In some embodiments, the sacrificial element and the triggering element together have the following structure:

[0318]

[0319] wherein the trigger element is represented by "peptide" and comprises one to ten amino acids, and Indicates the point of attachment to the payload. In some embodiments, the peptide includes Val-Cit or Val-Ala. Heterocyclic variants of this sacrificial element (eg, pyridinyl, pyrimidinyl, etc.) can also be used.

[0320] In some embodiments, the sacrificial element contains a phenolic group that is covalently attached to the remainder of the molecule via the phenolic oxygen. One such sacrificial element relies on an approach where diaminoethane "steric ligation" is used in conjunction with traditional "PABO" based sacrificial elements to deliver phenolics.

[0321] In some embodiments, the trigger element may comprise a non-cleavable portion or segment. Polyethylene glycol (PEG) and related polymers may be included with a cleavable group, such as a disulfide, hydrazone, or dipeptide, to form a sacrificial group and / or trigger element.

[0322] Other degradable bonds that can be included in the sacrificial element can include esters. Esters can be formed by the reaction of PEG carboxylic acid or activated PEG carboxylic acid with alcohol groups on the payload, and such ester groups can be hydrolyzed under physiological conditions to release the payload. Other hydrolytically degradable bonds can include carbonate bonds; imine bonds generated by the reaction of amines and aldehydes; phosphate bonds formed by the reaction of alcohols with phosphate groups; acetal bonds as the reaction product of aldehydes and alcohols; orthoester bonds as the reaction product of formate and alcohols; and oligonucleotide bonds formed by phosphoramidite groups (contained at the ends of the polymer) and the 5' hydroxyl group of the oligonucleotide.

[0323] In some embodiments, the trigger element, sacrificial group, and payload together have the following structure:

[0324]

[0325] in Indicates the attachment site to the remainder of the molecule (i.e., a compound of structure (I) or a conjugate of structure (II)), and the payload is indicated by text. In addition to other amino acids, the amino acids of the above embodiments may be substituted or used. In some embodiments, the trigger element is Asn-Cit, Arg-Cit, Val-Glu, Ser-Cit, Lys-Cit, Asp-Cit, Phe-Lys, Glu-Val-Cit, Glu-Val-Cit, Glu-Glu-Val-Cit, or Glu-Glu-Glu-Val-Cit, and the sacrificial element is PABC.

[0326] In some embodiments, the phenyl portion of PABC is substituted with one or more substituents. In some embodiments, the substituents have one of the following structures:

[0327]

[0328]

[0329] In some embodiments, the sacrificial group comprises one of the following structures:

[0330]

[0331] In some embodiments, the trigger element, sacrificial element, and payload together have one of the following structures, where the payload is represented by words:

[0332]

[0333] In some embodiments, the sacrificial element has the following structure:

[0334]

[0335]

[0336] The above structure shows a substitution pattern of 1, 3, 4 on the benzene ring of the sacrificial element. In some embodiments, the substitution pattern can be 1, 2, 4 (i.e., 1 is a bond to the payload, 2 is a bond to the rest of the molecule, and 4 is a bond to the carbohydrate) or 1, 3, 5 (i.e., 1 is a bond to the payload, 3 is a bond to the rest of the molecule, and 4 is a bond to the carbohydrate).

[0337] Although cleavable joints (e.g., joints with trigger elements or sacrificial elements) can provide certain advantages, joints do not have to be cleavable. For non-cleavable joints, payload release may not rely on different properties between plasma and some cytoplasmic compartments. The release of payload can occur after the conjugate of structure (II) is internalized and delivered to the lysosomal compartment by antigen-mediated endocytosis, in which the targeting moiety (or its binding fragment) can be degraded to the amino acid level by intracellular proteolysis. This process can release payload or payload derivatives. Payload or payload derivatives can be more hydrophilic and less membrane permeability, which can lead to less bystander effect and less non-specific toxicity compared to conjugates with cleavable joints. Conjugates with non-cleavable joints have higher stability in circulation than conjugates with cleavable joints. Non-cleavable joints can include alkylene chains, or can be polymerized, such as based on polyalkylene glycol polymers, amide polymers, or can include segmentation of alkylene chains, polyalkylene glycols and / or amide polymers. The linker may contain a polyethylene glycol segment having 1 to 6 ethylene glycol units. 1 -R 1 or L 2 -R 2 Includes linkers that are not cleavable in vivo.

[0338] In some embodiments, the trigger element and the sacrificial element together include one of the following structures:

[0339]

[0340]

[0341] In some embodiments, the heteroalkylene element comprises polyethylene glycol or polypropylene glycol. In some embodiments, the heteroalkylene element comprises one of the following structures:

[0342]

[0343]

[0344] in:

[0345] R 5b , R 5c , R 5d and R 5e is independently selected at each occurrence from the group consisting of hydrogen, deuterium, alkyl, haloalkyl, halo, alkoxy, haloalkoxy, amino, hydroxy, cyano, nitro, thiol, carboxyalkyl, alkyl-S(O) 3 H, Alkyl-OP(O) 3 H, Alkyl-P(O) 3 H, -O-carboxyalkyl, -O-alkyl-S(O) 3 H, -O-alkyl-OP(O) 3 H, -O-alkyl-P(O) 3 H, -S(O) 3 H, -OP(O) 3 H and -P(O) 3 H,

[0346] q1 is independently an integer from 1 to 24 at each occurrence.

[0347] In some embodiments, R 5b , R 5c , R 5d and R 5e It's all hydrogen.

[0348] In some embodiments, the polar cap includes one or more charged amino acids, one or more polyols, or a combination thereof. In certain embodiments, the polar cap includes diols, triols, tetraols, or a combination thereof. In some embodiments, the polar cap includes glycerol, trimethylolpropane, pentaerythritol, maltitol, sorbitol, xylitol, erythritol, isomalt, or a combination thereof. In certain embodiments, the polar cap includes one or more natural amino acids. In some embodiments, the polar cap includes one or more non-natural amino acids. In some embodiments, the polar cap includes one or more non-natural amino acids and one or more natural amino acids. In certain embodiments, the polar cap includes serine, threonine, cysteine, proline, asparagine, glutamine, lysine, arginine, histidine, aspartate, glutamate, 4-hydroxyproline, 5-hydroxylysine, homoserine, homocysteine, ornithine, β-alanine, statine, or γ-aminobutyric acid. In certain embodiments, the polar cap includes aspartic acid, serine, glutamic acid, serine-β-glucose, or a combination thereof.

[0349] In some embodiments, the polar cap comprises one of the following structures:

[0350]

[0351] In further embodiments, the polar cap has one of the following structures, including combinations thereof:

[0352]

[0353]

[0354] In some embodiments, L 1 , L 2 or L 3 A linker comprising: alkylene, alkylene-L a -, alkenylene, alkenylene-L a -, alkynylene, alkynylene-L a -、-L a -Alkylene-L a -、-L a -Alkenylene-L a -、-L a -Alkynylidene-L a - and combinations thereof, wherein each alkylene, alkenylene and alkynylene is optionally substituted, and L a is independently selected at each occurrence from -O-, -S-, -N(R 7 )-, -C(O)-, -C(S)-, -C(O)O-, -OC(O)-, -OC(O)O-, -C(O)N(R 7)-、-N(R 7 )C(O)-、-C(O)N(R 7 )C(O)-、-C(O)N(R 7 )C(O)N(R 7 )、-N(R 7 )C(O)N(R 7 )-、-N(R 7 )C(O)O-、-OC(O)N(R 7 )-、-C(NR 7 )-、-N(R 7 )C(NR 7 )-、-C(NR 7 )N(R 7 )-、-N(R 7 )C(NR 7 )N(R 7 )-、-S(O) 2 -, -OS(O)-, -S(O)O-, -S(O), -OS(O) 2 -、-S(O) 2 O、-N(R 7 )S(O) 2 -、-S(O) 2 N(R 7 )-、-N(R 7 )S(O)-、-S(O)N(R 7 )-、-N(R 7 )S(O) 2 N(R 7 )- and -N(R 7 )S(O)N(R 7 )-, and R 7 is independently selected at each occurrence from hydrogen, -NH 2 、-C(O)OCH 2 C 6 H 5 ; and C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-12 Cycloalkyl and 3 to 12 membered heterocyclic rings, each of which is optionally substituted with substituents independently selected from the group consisting of halo, hydroxy, cyano, nitro, amino, oxo, thio, -C(O)OCH 2 C 6 H 5 、-NHC(O)OCH 2 C 6 H 5 , C 1-10 Alkyl, C 1-10Haloalkyl, C 1-10 Alkoxy, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-12 Cycloalkyl and 3- to 12-membered heterocyclyl.

[0355] In some embodiments, each L 1 , L 2 or L 3 Optionally substituted with one or more substituents selected from the group consisting of alkyl, alkenyl, alkynyl, halo, hydroxy, cyano, -OR 8 、-SR 8 , amino, amine, acylamino, cycloalkyl, aryl, heterocyclyl, heteroaryl, cycloalkylalkyl, arylalkyl, heterocycloalkylalkyl, heteroarylalkyl, -C(O)R 8 、-C(O)N(R 8 ) 2 、-N(R 8 )C(O)R 8 、-C(O)OR 8 、-OC(O)R 8 、-S(O)R 8 、-S(O) 2 R 8 、-P(O)(OR 8 ) 2 、-OP(O)(OR 8 ) 2 , nitro, oxo, thio, =N(R 8 ) or cyano, and R 8 is hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heterocyclyl, heteroaryl, cycloalkylalkyl, arylalkyl, heterocycloalkylalkyl or heteroarylalkyl.

[0356] In some embodiments, L 1 , L 2 or L 3 Independently selected from the following structures:

[0357]

[0358] in:

[0359] R a is hydrogen or alkyl;

[0360] L b is independently at each occurrence a direct bond, an optionally substituted alkylene linker, an optionally substituted heteroalkylene linker, a heteroatom linker, or a combination thereof; and L c is independently at each occurrence an optionally substituted alkylene linker, and provided that L 1 , L2 or L 3 At least one of has the following structure:

[0361]

[0362] In some embodiments, L 1 and L 2 Independently selected from the following structures:

[0363]

[0364] in:

[0365] R a is hydrogen or alkyl;

[0366] L b is independently at each occurrence a direct bond, an optionally substituted alkylene linker, an optionally substituted heteroalkylene linker, a heteroatom linker, or a combination thereof;

[0367] L c is independently at each occurrence an optionally substituted alkylene linker;

[0368] The condition is L 1 or L 2 At least one of has the following structure;

[0369]

[0370] In further embodiments, L 2 Has the following structure:

[0371]

[0372] In some embodiments, L c is unsubstituted. In some embodiments, L c It is C 1 -C 6 In some more embodiments, L c It is C 2 -C 4 In some embodiments, L c It is a straight chain C 1 -C 6 In more embodiments, L c It is a straight chain unsubstituted C 1 -C 6 In more embodiments, L c It is a straight chain unsubstituted C 2 -C 4 Alkylene.

[0373] In further embodiments, L 1 , L 2 and L 3 Each independently has one of the following structures:

[0374]

[0375] in:

[0376] *Indicates a direct bond to a substitutable position on the phenyl group of the following structure:

[0377]

[0378] In some embodiments, L 1 or L 2 Has one of the following structures:

[0379]

[0380] in:

[0381] *Indicates a direct bond to a substitutable position on the phenyl group of the following structure:

[0382]

[0383] In some embodiments, L c Substituted with one or more substituents selected from the group consisting of halo, haloalkyl, alkoxy, cyano, nitro, carboxyl, sulfonamide, sulfonic acid, or a combination thereof.

[0384] In some embodiments, the compound has one of the following structures (Ia) or (Ib):

[0385]

[0386] In some embodiments, the compound has one of the following structures (Ic') or (Ic"):

[0387]

[0388] In certain embodiments, X 2 , X 3 or both are CH or CF. In some embodiments, X 1 , X 5 or both are CR 3 , and R 3 is H or halo. In some embodiments, X 1 and X 5 All are CR 3 , and R3 is H or halo. In some embodiments, X 1 Yes CR 3 , and R 3 In certain embodiments, X 5 Yes CR 3 , and R 3 In some embodiments, halo is fluoro. In some embodiments, X 1 In some embodiments, X 1 is CH. In some embodiments, X 5 is CH. In some embodiments, X 5 In some embodiments, X 3 Yes CR 3 , and R 3 is H. In some embodiments, X 3 Yes CR 3 , and R 3 is halogenated (eg, fluorinated).

[0389] In some embodiments, the compound has one of the following structures: (Ia-1), (Ia-2), (Ia-3), (Ia-4), (Ia-5), or (Ia-6):

[0390]

[0391]

[0392] in:

[0393] L b is independently at each occurrence a direct bond, an optionally substituted alkylene linker, an optionally substituted heteroalkylene linker, a heteroatom linker, or a combination thereof; and

[0394] q6 is 0, 1 or 2.

[0395] In some embodiments, the compound has one of the following structures: (Ia-1), (Ia-2), (Ia-3), (Ia-4), (Ia-5), (Ia-6), (Ia-7), or (Ia-8):

[0396]

[0397]

[0398] in:

[0399] L bis independently at each occurrence a direct bond, an optionally substituted alkylene linker, an optionally substituted heteroalkylene linker, a heteroatom linker, or a combination thereof; and

[0400] q6 is 0, 1 or 2.

[0401] In some embodiments, q6 is 1, and L b It's gly-gly.

[0402] In certain embodiments, the compound has one of the following structures: (Ic-1), (Ic-2), (Ic-3), or (Ic-4):

[0403]

[0404]

[0405] in:

[0406] L b is independently at each occurrence a direct bond, an optionally substituted alkylene linker, an optionally substituted heteroalkylene linker, a heteroatom linker, or a combination thereof; and

[0407] q7 is 1, 2 or 3.

[0408] In some embodiments, q7 is 2.

[0409] In some embodiments, the compound has the following structure (Id), (Ie), (If), or (Ig):

[0410]

[0411] in:

[0412] L b is independently at each occurrence a direct bond, an optionally substituted alkylene linker, an optionally substituted heteroalkylene linker, a heteroatom linker, or a combination thereof;

[0413] q8 is 0, 1, or 2; and

[0414] q9 is 0, 1, or 2.

[0415] In some embodiments, q9 is 0 and q8 is 1.

[0416] In some embodiments, the compound has one of the following structures (Ih) or (Ii):

[0417]

[0418] in:

[0419] Lb is independently at each occurrence a direct bond, an optionally substituted alkylene linker, an optionally substituted heteroalkylene linker, a heteroatom linker, or a combination thereof.

[0420] In some embodiments, L b is a direct bond, an optionally substituted alkylene linker, or an optionally substituted heteroalkylene linker.

[0421] In some embodiments, L b is a direct key or has one of the following structures:

[0422]

[0423] in:

[0424] R b is independently at each occurrence hydrogen, alkyl, hydroxyalkyl or alkoxyalkyl.

[0425] In some embodiments, R b In each occurrence is –CH 3 .

[0426] In some embodiments, L 1 or L 2 Has the following structure:

[0427]

[0428] in:

[0429] ** Shown with X 1 , X 2 , X 3 , X 4 or X 5 key.

[0430] In certain embodiments, X 2 It is CL 1 -R 1 , and X 3 It is CL 2 -R 2 In some embodiments, X 3 It is CL 1 -R 1 , and X 2 It is CL 2 -R 2 In some embodiments, X 1 , X 4 and X 5 All CR 3 In some embodiments, X 1 , X 4and X 5 All are CH.

[0431] In some embodiments, the compound has one of the following structures:

[0432]

[0433]

[0434]

[0435] in:

[0436] R 1b is a chemotherapeutic, cytotoxic, or myeloid cell agonist; R 2b Has one of the following structures:

[0437]

[0438]

[0439] L 1g Has one of the following structures:

[0440]

[0441]

[0442] In some embodiments, the compound has one of the following structures:

[0443]

[0444]

[0445]

[0446] in:

[0447] R 1b is a chemotherapeutic agent, cytotoxic agent, or myeloid cell agonist; R 2b Has one of the following structures:

[0448]

[0449] L 1g Has one of the following structures:

[0450]

[0451]

[0452] In some embodiments, the compound has one of the following structures:

[0453]

[0454] In some embodiments, the compound has one of the following structures:

[0455]

[0456]

[0457]

[0458] In some embodiments, the compound has one of the following structures:

[0459]

[0460]

[0461]

[0462]

[0463]

[0464]

[0465]

[0466]

[0467] In some embodiments, the compound has one of the following structures:

[0468]

[0469] As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0470] In some embodiments, the compound has one of the following structures:

[0471]

[0472]

[0473]

[0474]

[0475]

[0476]

[0477]

[0478]

[0479] As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0480] In some embodiments, the payload is a chemotherapeutic agent, a cytotoxic agent, or a myeloid cell agonist.

[0481] In some embodiments, the payload is a chemotherapeutic agent selected from the group consisting of camptothecin, paclitaxel, doxorubicin, vinblastine, dacarbazine, irinotecan, topotecan, xinotecan, cositect, exitecan, lortotecan, SN-38, Dxd (i.e., CAS No. 1599440-33-1), gimatitecan, belotecan, and rubitecan.

[0482] In certain embodiments, the payload is a cytotoxic agent selected from the group consisting of calicheamicin, anthramycin, abemycin, cheikamycin, DC-81, methylaminomycin, neoaminomycin A, neoaminomycin B, puromycin, prolactin, sibamicin, sibirinomycin, tomamycin, auristatin F, monomethyl auristatin F, auristatin E, monomethyl auristatin E, dolestatin, monomethyl dolestatin, maytansine, and maytansine.

[0483] In certain embodiments, the payload is a myeloid cell agonist selected from the group consisting of a STING agonist, a ligand of TLR2, a ligand of TLR3, a ligand of TLR4, a ligand of TLR5, a ligand of TLR6, a ligand of TLR7, a ligand of TLR8, a ligand of TLR9, a ligand of TLR10, a ligand of a nucleotide oligomerization domain (NOD), a ligand of a RIG-I-like receptor (RLR), a ligand of a C-type lectin receptor (CLR), a cytoplasmic DNA sensor The invention relates to a ligand of a CD4+ / CD5+ receptor (CDS) and a ligand of an inflammasome inducer, preferably wherein the myeloid cell agonist is selected from the group consisting of sergantomod, motomod, resiquimod, 3M-051, 3M-052, MCT-465, IMO-4200, VTX-763, VTX-1463, RG7854, ADU-S100, MK-1454, MK-2118, BMS-986301, GSK3745417, SB-11285 and IMSA-101.

[0484] In some embodiments, the payload is an alkylating agent, an antimetabolite, a microtubule inhibitor, a topoisomerase inhibitor, a myeloid agonist, a glucocorticoid receptor agonist, or a cytotoxic antibiotic. In certain embodiments, the payload is a nitrogen mustard, a nitrosourea, a tetrazine, an aziridine, cisplatin or a cisplatin derivative, or a non-classical alkylating agent.

[0485] In certain embodiments, the payload is nitrogen mustard, cyclophosphamide, melphalan, chlorambucil, ifosfamide, busulfan, N-nitroso-N-methylurea (MNU), carmustine (BCNU), lomustine (CCNU), semustine (MeCCNU), fotemustine, streptozotocin, dacarbazine, mitozolomide, temozolomide, thiotepa, mitomycin, diazinon (AZQ), cisplatin, carboplatin, oxaliplatin, procarbazine or hexamethylmelamine. In some embodiments, the payload is antifolate, fluoropyrimidine, deoxynucleoside analog or thiopurine.

[0486] In certain embodiments, the payload is methotrexate, pemetrexed, fluorouracil, capecitabine, cytarabine, gemcitabine, decitabine, azacitidine, fludarabine, nelarabine, cladribine, clofarabine, pentostatin, thioguanine and mercaptopurine. In some embodiments, the payload is auristatin, a vinca alkaloid or a taxane. In certain embodiments, the payload is auristatin F, auristatin E, vincristine, vinblastine, vinorelbine, vindesine, vinflunine, paclitaxel, docetaxel, etoposide or teniposide. In other embodiments, the payload is hydrocortisone (e.g., prednisone, fluocinolone), acetonide (e.g., budesonide, fluocinolone), methotrexate (e.g., dexamethasone, betamethasone, fluticasone).

[0487] In some embodiments, the molecular weight of the payload is greater than 150 g / mole, greater than 200 g / mole, greater than 300 g / mole, greater than 400 g / mole, greater than 500 g / mole, greater than 600 g / mole, greater than 700 g / mole, greater than 800 g / mole, greater than 900 g / mole, or greater than 1000 g / mole. In some embodiments, the molecular weight of the payload is less than 150 g / mole, less than 200 g / mole, less than 300 g / mole, less than 400 g / mole, less than 500 g / mole, less than 600 g / mole, less than 700 g / mole, less than 800 g / mole, less than 900 g / mole, or less than 1000 g / mole.

[0488] The payload portion is known in the art. For example, the payload can be found in PCT Publication Nos. WO2021 / 207701, WO 2019 / 217591, WO 2018 / 089373, WO 2019 / 136487, and U.S. Pat. No. 11,179,473, and references cited therein, and the payloads from each of the references are incorporated herein by reference in their entirety.

[0489] In some embodiments, the payload is a STING agonist having the following structure (III):

[0490]

[0491] in:

[0492] L 1 is a divalent linker having a single bond to X 1 The first end is covalently bound to X via a single bond 2 The second end of the covalent bond, where L 1 It is C 2 -C 8 Alkylene, C 2 -C 8 Alkenylene, C 2 -C 8 Alkynylidene, C 2 -C 8 Heteroalkylene, C 2 -C 8 Heteroalkenylene, C 2 -C 8 Heteroalkynylene or -(L 1a ) x -L 1b -(L 1c ) y -, each of which is optionally substituted by one or more R, wherein:

[0493] L 1a and L 1c Each independently is C 1 -C 6 Alkylene, C 1 -C 6 Alkenylene, C 2 -C 6 Alkynylidene, C 1 -C 6 Heteroalkylene, C 2 -C 6 Heteroalkenylene, C 2 -C 6 heteroalkynylene, each of which is optionally substituted with one or more R;

[0494] L 1b It is C 6 -C 10 Arylene, C 3 -C 6 cycloalkylene, 3-6 membered heterocyclylene or 5-6 membered heteroarylene, each of which is optionally substituted with one or more R; and

[0495] x and y are each independently 0 or 1;

[0496] R is independently R*, D, -OR at each occurrence a 、-N(R a ) 2 、-NR a C(=O)R b 、-C(=O)N(R a ) 2 , -CO 2 R a 、-OC(=O)R b 、-OC(=O)N(R a ) 2 、-OC(=O)N(R a ) 2 、-SR a 、-S(O) 2 N(R a ) 2 、-S(O) 2 R a , C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, C 6 -C 10 Aryl, C 3 -C 6 Cycloalkyl, 3-6 membered heterocyclyl or 5-6 membered heteroaryl, wherein each C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, C 6 -C 10 Aryl, C 3 -C 6 Cycloalkyl, 3-6 membered heterocyclyl and 5-6 membered heteroaryl are independently optionally substituted by one or more substituents selected from the following: D, -OR a 、-N(R a ) 2 、-NR a C(=O)R b 、-C(=O)N(Ra ) 2 , -CO 2 R a 、-OC(=O)R b 、-OC(=O)N(R a ) 2 、-OC(=O)N(R a ) 2 、-SR a 、-S(=O) 2 N(R a ) 2 or -S(O) 2 R a ;

[0497] X 1 and X 2 Each independently is O or NR a ;

[0498] (i)W 1 and W 2 Independently OR c or SR c ;

[0499] Y 1 and Y 2 Each is N;

[0500] R 1 It is H, CN, OH, Cl, Br, N 3 , C 1 -C 6 Alkyl or C 1 -C 6 alkoxy; and

[0501] R 2 It is H, CN, OH, Cl, Br, N 3 , C 1 -C 6 Alkyl or C 1 -C 6 Alkoxy;

[0502] (ii)W 1 and W 2 Independently OR c , SR c or BHR c ;

[0503] Y 1 and Y 2 At least one of is CH, CF or CCl, and Y 1 and Y 2 the other of which is CH, CF, CCl or N; and

[0504] R 1 and R 2 are independently H, CN, OH, Cl, Br, N 3 , C 1 -C 6 Alkyl or C 1 -C 6 Alkoxy; or

[0505] (iii)W 1 and W 2 Independently SR c or BHR c ;

[0506] Y 1 and Y 2 each is N; and

[0507] R 1 It is H, CN, Cl, Br, N 3 , C 1 -C 6 Alkyl or C 1 -C 6 alkoxy; and

[0508] R 2 It is H, CN, OH, F, Cl, Br, N 3 , C 1 -C 6 Alkyl or C 1 -C 6 Alkoxy;

[0509] R 3 and R 4 are each independently H or -N(R a ) 2 ;

[0510] R a In each occurrence, it is independently H, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 1 -C 6 Aminoalkyl, C 2 -C 6 Amine alkenyl, C 3 -C 6 Cycloalkyl, C 3 -C 6 Aminocycloalkyl, C 3 -C 6 Cycloalkyl-C 1 -C 6 Alkyl, C3 -C 6 Aminocycloalkyl-C 1 -C 6 Alkyl, 3-6 membered heterocyclic group, 3-6 membered amino heterocyclic group, 3-6 membered heterocyclic group-C 1 -C 6 Alkyl or 3-6 membered amino heterocyclic group-C 1 -C 6 Alkyl, wherein each substitutable R a Optionally substituted with R*;

[0511] R b C independently at each occurrence 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 1 -C 6 Aminoalkyl, C 2 -C 6 Amine alkenyl, C 3 -C 6 Cycloalkyl, C 3 -C 6 Aminocycloalkyl, C 3 -C 6 Cycloalkyl-C 1 -C 6 Alkyl, C 3 -C 6 Aminocycloalkyl-C 1 -C 6 Alkyl, 3-6 membered heterocyclic group, 3-6 membered amino heterocyclic group, 3-6 membered heterocyclic group-C 1 -C 6 Alkyl or 3-6 membered amino heterocyclic group-C 1 -C 6 Alkyl, where each R b Optionally substituted with R*;

[0512] R c In each occurrence, it is independently H, C 1 -C 6 Alkyl, -CH 2 OC(=O)R d or -R e R f , where each substitutable R c Optionally substituted with R*;

[0513] R d C independently at each occurrence 1 -C 6 Alkyl, where each R d Optionally substituted with R*;

[0514] R e Independently absent at each occurrence, C 1 -C 6 Alkylene or C 1 -C 6 wherein each alkylene and heteroalkylene is optionally substituted by one or more R g replace;

[0515] R f In each occurrence, it is independently H, C 1 -C 20 Alkyl, C 1 -C 20 Heteroalkyl, -OC(O)OC 1 -C 20 alkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein each alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally substituted by one or more R h replace;

[0516] R g is independently at each occurrence R*, halo, -CN, C 1 -C 20 Alkyl, -OR i , oxo, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein each alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally replaced by one or more R j replace;

[0517] R h R*, C independently at each occurrence 1 -C 20 Alkyl, C 1 -C 20 Heteroalkyl, -C(=O)C 1 -C 20 Alkyl, -OC(=O)C 1 -C 20 Alkyl, -C(=O)OC 1 -C 20 Alkyl, -OC(=O)OC 1 -C 20 Alkyl, -C(=O)N(R k )C 1 -C 20 Alkyl, -N(R k )C(=O)C 1 -C 20 Alkyl, -OC(=O)N(R k )C 1 -C 20 Alkyl, -C(=O)C1 -C 20 Heteroalkyl, -OC(=O)C 1 -C 20 Heteroalkyl, -C(=O)OC 1 -C 20 Heteroalkyl, -OC(=O)OC 1 -C 20 heteroalkyl, -C(=O)N(R k )C 1 -C 20 Heteroalkyl, -N(R k )C(=O)C 1 -C 20 heteroalkyl, -OC(=O)N(R k )C 1 -C 20 heteroalkyl, -Oaryl, -Oheteroaryl, -C(=O)aryl, -C(=O)heteroaryl, -OC(=O)aryl, -C(=O)Oaryl, -OC(=O)heteroaryl, -C(=O)Oheteroaryl, -C(=O)N(R k )aryl, -C(=O)N(R k )heteroaryl, -N(R k )C(=O)aryl, -N(R k )C(=O)heteroaryl, -OC(=O)N(R k )aryl, -OC(=O)N(R k )heteroaryl, -N(R k )C(=O)Oaryl, -N(R k )C(=O)Oheteroaryl, -S(=O) 2 N(R k )aryl, -S(=O) 2 N(R k )heteroaryl or -N(R k )-*Peptide-L 2 , where * is the C-terminus of the peptide, and L 2 Does not exist, -C 1 -C 12 Alkyl, -C 1 -C 12 Heteroalkyl, -C(=O)C 1 -C 12 Alkyl or -C(=O)C 1 -C 12 wherein each alkyl, heteroalkyl, aryl and heteroaryl is optionally substituted by one or more R j replace;

[0518] R i In each occurrence, it is independently H, C1 -C 20 Alkyl, C 1 -C 20 heteroalkylcycloalkyl, heterocyclyl, aryl or heteroaryl, wherein each alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally substituted by one or more R j replace;

[0519] R j R*, C independently at each occurrence 1 -C 20 Alkyl, -OC 1 -C 20 Alkyl, C 1 -C 20 heteroalkyl, halo, -CN, -OH, oxo, aryl, heteroaryl, -Oaryl, or -Oheteroaryl;

[0520] R k is H or C independently at each occurrence 1 -C 20 Alkyl; and

[0521] R* is a divalent linker having a first end covalently bonded to the remainder of structure (I) or structure (II) via a single bond and a second end covalently bonded to L via a single bond. 1 , X 1 , X 2 , R 3 , R 4 , W 1 or W 2 At least one of is covalently bound and replaced by R*.

[0522] In other embodiments, the STING agonist has the following structure (IIIA):

[0523]

[0524] In still different embodiments, the STING agonist is a compound having the following structure (IIIA'):

[0525]

[0526] In other embodiments, the present disclosure provides a compound having activity as a STING agonist and having the following structure (IV):

[0527]

[0528] or a pharmaceutically acceptable salt, tautomer, prodrug or stereoisomer thereof, wherein:

[0529] L 1is a divalent linker having a single bond to X 1 The first end is covalently bound to X via a single bond 2 The second end of the covalent bond, where L 1 It is C 2 -C 8 Alkylene, C 2 -C 8 Alkenylene, C 2 -C 8 Alkynylidene, C 2 -C 8 Heteroalkylene, C 2 -C 8 Heteroalkenylene, C 2 -C 8 Heteroalkynylene or -(L 1a ) x -L 1b -(L 1c ) y -, each of which is optionally substituted by one or more R, wherein:

[0530] L 1a and L 1c Each independently is C 1 -C 6 Alkylene, C 1 -C 6 Alkenylene, C 2 -C 6 Alkynylidene, C 1 -C 6 Heteroalkylene, C 2 -C 6 Heteroalkenylene, C 2 -C 6 heteroalkynylene, each of which is optionally substituted with one or more R;

[0531] L 1b It is C 6 -C 10 Arylene, C 3 -C 6 cycloalkylene, 3-6 membered heterocyclylene or 5-6 membered heteroarylene, each of which is optionally substituted with one or more R; and

[0532] x and y are each independently 0 or 1;

[0533] R is independently R*, D, -OR at each occurrence a 、-N(R a ) 2 、-NR a C(=O)R b 、-C(=O)N(R a) 2 , -CO 2 R a 、-OC(=O)R b 、-OC(=O)N(R a ) 2 、-OC(=O)N(R a ) 2 、-SR a 、-S(O) 2 N(R a ) 2 、-S(O) 2 R a , C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, C 6 -C 10 Aryl, C 3 -C 6 Cycloalkyl, 3-6 membered heterocyclyl or 5-6 membered heteroaryl, wherein each C 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, C 6 -C 10 Aryl, C 3 -C 6 Cycloalkyl, 3-6 membered heterocyclyl and 5-6 membered heteroaryl are independently optionally substituted by one or more substituents selected from the following: D, -OR a 、-N(R a ) 2 、-NR a C(=O)R b 、-C(=O)N(R a ) 2 , -CO 2 R a 、-OC(=O)R b 、-OC(=O)N(R a ) 2 、-OC(=O)N(R a ) 2 、-SR a 、-S(=O) 2 N(R a ) 2 or -S(O) 2 R a ;

[0534] W 1 and W 2 Each independently is OR c , SRc or BHR c ;

[0535] X 1 and X 2 Each independently is O or NR a ;

[0536] (i)Y 1 and Y 2 Each is N;

[0537] R 5 It is H, CN, OH, Cl, Br, N 3 , C 1 -C 6 Alkyl or C 1 -C 6 alkoxy; and

[0538] R 6 It is H, CN, F, Cl, Br, N 3 , C 1 -C 6 Alkyl or C 1 -C 6 Alkoxy;

[0539] or

[0540] (ii) Y 1 and Y 2 At least one of is CH, CF or CCl, and Y 1 and Y 2 the other of which is CH, CF, CCl or N; and

[0541] R 5 and R 6 are independently H, CN, OH, Cl, Br, N 3 , C 1 -C 6 Alkyl or C 1 -C 6 Alkoxy;

[0542] R 3 and R 4 are each independently H or -N(R a ) 2 ;

[0543] R a In each occurrence, it is independently H, C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 1 -C 6 Aminoalkyl, C2 -C 6 Amine alkenyl, C 3 -C 6 Cycloalkyl, C 3 -C 6 Aminocycloalkyl, C 3 -C 6 Cycloalkyl-C 1 -C 6 Alkyl, C 3 -C 6 Aminocycloalkyl-C 1 -C 6 Alkyl, 3-6 membered heterocyclic group, 3-6 membered amino heterocyclic group, 3-6 membered heterocyclic group-C 1 -C 6 Alkyl or 3-6 membered amino heterocyclic group-C 1 -C 6 Alkyl, wherein each substitutable R a Optionally substituted with R*;

[0544] R b C independently at each occurrence 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 1 -C 6 Aminoalkyl, C 2 -C 6 Amine alkenyl, C 3 -C 6 Cycloalkyl, C 3 -C 6 Aminocycloalkyl, C 3 -C 6 Cycloalkyl-C 1 -C 6 Alkyl, C 3 -C 6 Aminocycloalkyl-C 1 -C 6 Alkyl, 3-6 membered heterocyclic group, 3-6 membered amino heterocyclic group, 3-6 membered heterocyclic group-C 1 -C 6 Alkyl or 3-6 membered amino heterocyclic group-C 1 -C 6 Alkyl, where each R b Optionally substituted with R*;

[0545] R c In each occurrence, it is independently H, C 1 -C 6 Alkyl, -CH 2 OC(=O)R d or -Re R f , where each substitutable R c Optionally substituted with R*;

[0546] R d C independently at each occurrence 1 -C 6 Alkyl, where each R d Optionally substituted with R*;

[0547] R e Independently absent at each occurrence, C 1 -C 6 Alkylene or C 1 -C 6 wherein each alkylene and heteroalkylene is optionally substituted by one or more R g replace;

[0548] R f In each occurrence, it is independently H, C 1 -C 20 Alkyl, C 1 -C 20 Heteroalkyl, -OC(O)OC 1 -C 20 alkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein each alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally substituted by one or more R h replace;

[0549] R g is independently at each occurrence R*, halo, -CN, C 1 -C 20 Alkyl, -OR i , oxo, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein each alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally replaced by one or more R j replace;

[0550] R h R*, C independently at each occurrence 1 -C 20 Alkyl, C 1 -C 20 Heteroalkyl, -C(=O)C 1 -C 20 Alkyl, -OC(=O)C 1 -C 20 Alkyl, -C(=O)OC 1 -C 20 Alkyl, -OC(=O)OC 1 -C 20Alkyl, -C(=O)N(R k )C 1 -C 20 Alkyl, -N(R k )C(=O)C 1 -C 20 Alkyl, -OC(=O)N(R k )C 1 -C 20 Alkyl, -C(=O)C 1 -C 20 Heteroalkyl, -OC(=O)C 1 -C 20 Heteroalkyl, -C(=O)OC 1 -C 20 Heteroalkyl, -OC(=O)OC 1 -C 20 heteroalkyl, -C(=O)N(R k )C 1 -C 20 Heteroalkyl, -N(R k )C(=O)C 1 -C 20 heteroalkyl, -OC(=O)N(R k )C 1 -C 20 heteroalkyl, -Oaryl, -Oheteroaryl, -C(=O)aryl, -C(=O)heteroaryl, -OC(=O)aryl, -C(=O)Oaryl, -OC(=O)heteroaryl, -C(=O)Oheteroaryl, -C(=O)N(R k )aryl, -C(=O)N(R k )heteroaryl, -N(R k )C(=O)aryl, -N(R k )C(=O)heteroaryl, -OC(=O)N(R k )aryl, -OC(=O)N(R k )heteroaryl, -N(R k )C(=O)Oaryl, -N(R k )C(=O)Oheteroaryl, -S(=O) 2 N(R k )aryl, -S(=O) 2 N(R k )heteroaryl or -N(R k )-*Peptide-L 2 , where * is the C-terminus of the peptide, and L 2 Does not exist, -C 1 -C 12 Alkyl, -C 1 -C 12Heteroalkyl, -C(=O)C 1 -C 12 Alkyl or -C(=O)C 1 -C 12 wherein each alkyl, heteroalkyl, aryl and heteroaryl is optionally substituted by one or more R j replace;

[0551] R i In each occurrence, it is independently H, C 1 -C 20 Alkyl, C 1 -C 20 heteroalkylcycloalkyl, heterocyclyl, aryl or heteroaryl, wherein each alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally substituted by one or more R j replace;

[0552] R j R*, C independently at each occurrence 1 -C 20 Alkyl, -OC 1 -C 20 Alkyl, C 1 -C 20 heteroalkyl, halo, -CN, -OH, oxo, aryl, heteroaryl, -Oaryl, or -Oheteroaryl;

[0553] R k is H or C independently at each occurrence 1 -C 20 Alkyl; and

[0554] R* is a divalent linker having a first end covalently bonded to the remainder of structure (I) or structure (II) via a single bond and a second end covalently bonded to L via a single bond. 1 , X 1 , X 2 , R 3 , R 4 , W 1 or W 2 At least one of is covalently bound and replaced by R*.

[0555] In other embodiments, the STING agonists of the present disclosure have the following structure (IVA):

[0556]

[0557] In other embodiments, the STING agonist has the following structure (IVA'):

[0558]

[0559] In some embodiments, R 1a Has one of the following structures:

[0560]

[0561]

[0562] In some embodiments, R 1a Has one of the following structures:

[0563]

[0564]

[0565]

[0566] In some embodiments, R 1a Has one of the following structures:

[0567]

[0568]

[0569]

[0570] In some embodiments, the payload or R 1a Has one of the following structures:

[0571]

[0572]

[0573] in:

[0574] R' is hydrogen or has one of the following structures:

[0575]

[0576] in:

[0577] R a ' is H or C 1-6 alkyl;

[0578] R b 'It's C 1-6 Alkyl or C 1-6 Alkoxy;

[0579] R c 'It is H, C 1-6 Alkyl, -CH 2 OH or C 1-6 Alkoxy;

[0580] R d ' is H or C 1-6 Alkyl; or

[0581] R e ' is H or C 1-6 alkyl.

[0582] In some embodiments, the payload has the following structure:

[0583]

[0584] In some embodiments, the payload has the following structure:

[0585]

[0586] In some embodiments, the payload has the following structure:

[0587]

[0588] In certain embodiments, the payload has the following structure:

[0589]

[0590] In some embodiments, the payload has the following structure:

[0591]

[0592] In certain embodiments, the payload has the following structure:

[0593]

[0594] In some embodiments, the payload has the following structure:

[0595]

[0596] In certain embodiments, the payload has the following structure:

[0597]

[0598] In some embodiments, the payload has the following structure:

[0599]

[0600] In certain embodiments, the payload has the following structure:

[0601]

[0602] In some embodiments, the payload has the following structure:

[0603]

[0604] In certain embodiments, the payload has the following structure:

[0605]

[0606] In certain embodiments, the payload has the following structure:

[0607]

[0608] In some embodiments, the payload has the following structure:

[0609]

[0610] In certain embodiments, the payload has the following structure:

[0611]

[0612] In some embodiments, the payload has the following structure:

[0613]

[0614] In certain embodiments, the payload has the following structure:

[0615]

[0616] A specific embodiment provides a compound having a structure selected from Table 1 (below), as a stereoisomer, enantiomer or tautomer thereof, or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0617]

[0618]

[0619]

[0620]

[0621]

[0622]

[0623]

[0624]

[0625]

[0626]

[0627]

[0628]

[0629]

[0630]

[0631]

[0632]

[0633]

[0634]

[0635]

[0636]

[0637]

[0638]

[0639]

[0640]

[0641]

[0642]

[0643]

[0644]

[0645]

[0646]

[0647]

[0648]

[0649]

[0650]

[0651]

[0652]

[0653]

[0654]

[0655]

[0656] The characterization data of compounds I-35-55 and I-57-62 are as follows:

[0657] Compound I-35

[0658] MS m / z[MH] - (ESI): 1977.90; 1 H NMR (300 MHz, methanol-d 4 )δ0.79(d,J=7.1Hz,3H),0.89(s,3H),1.08(t,J=7.1Hz,2H),1.13-1.3 7(m,7H),1.47(s,4H),1.62-1.76(m,3H),1.73-1.89(m,1H),2.11(d,J= 12.7Hz,3H),2.24(t,J=7.2Hz,3H),2.31-2.45(m,2H),2.49-2.62(m,2H ),2.78-3.16(m,4H),3.28(d,J=5.4Hz,2H),3.40-3.48(m,2H),3.48-3. 58(m,36H),3.56-3.74(m,6H),3.79(d,J=11.7Hz,3H),3.98(s,2H),4. 15-4.30(m,2H),4.30-4.45(m,2H),4.55-4.77(m,4H),4.95(d,J=4.3Hz ,1H),5.39(s,1H),5.51(s,1H),6.13-6.24(m,2H),6.88(s,1H),7.01(t ,J=8.9Hz,1H),7.07-7.25(m,7H),7.23-7.32(m,1H),7.32-7.49(m,2H)

[0659] Compound I-36

[0660] MS m / z[MH] - (ESI): 1834.50; 1H NMR (300 MHz, methanol-d 4 )δ:0.75-1.12(m,3H),1.20-1.43(m,1H),1.51-1.68(m,4H),1.70-1.84(m,2H),1.85-2.06(m,2H),2.05-2.42(m,6H),2.45-2 .61(m,4H),2.69(d,J=31.9Hz,1H),3.08(s,2H),3.41(t,J=5.7Hz,2H),3.53-3.59(m,2H),3.59-3.66(m,43H),3.74(t,J=6.2H z,2H),3.90(d,J=21.1Hz,4H),4.11(d,J=4.1Hz,2H),4.23(dd,J=7.9Hz,4.2Hz,1H),4.32(d,J=9.7Hz,1H),4.92-5.16(m,4H), 5.34-5.54(m,1H),5.60(s,1H),5.90-6.03(m,1H),6.26-6.63(m,3H),6.82-7.46(m,6H),7.56-7.66(m,1H),7.73-7.88(m,1H)

[0661] Compound I-37

[0662] MS m / z[MH] - (ESI): 1801.60; 1 H NMR (300 MHz, methanol-d 4 )δ:0.98(s,3H),1.27-1.34(m,6H),1.57(s,4H),1.80-1.91(m,4H),2.10-2.20(m,2H),2.37-2.39(m,4 H),2.63-2.70(m,5H),3.13-3.20(m,2H),3.37-3.40(m,3H),3.52-3.59(m,6H),3.61-3.62(m,37H),3.7 2-3.74(m,2H),3.92(s,3H),4.09(s,2H),4.17-4.23(m,3H),4.26-4.43(m,2H),4.66-4.75(m,4H),5.03 -5.04(m,1H),5.50-5.58(m,2H),6.31-6.34(m,2H),6.97(s,2H),7.10-7.20(m,2H),7.32-7.47(m,6H).

[0663] Compound I-38

[0664] MS m / z[MH] - (ESI): 977.30; 1 H NMR (300 MHz, methanol-d 4 )0.99(s,3H),1.56 -1.60(m,4H),1.76-1.78(m,2H),1.91(d,J=13.4Hz,1H),2.31-2.34(m,3H),2. 49-2.53(m,2H),2.84-2.93(m,3H),3.41-3.44(m,2H),3.80(s,3H),4.02-4.04( m,2H),4.30-4.33(m,1H),4.86-4.99(m,1H),5.02-5.05(m,2H),5.50-5.59(m,2 H),6.30-6.34(m,3H),6.81-6.93(m,3H),7.01-7.21(m,3H),7.33-7.38(m,3H).

[0665] Compound I-39

[0666] MS m / z[MH] - (ESI): 949.10; 1 H NMR (300 MHz, methanol-d 4 )δ:0.99(s,3H),1.20-1.30(m,1H),1.57(s,4H),1.70-1.78(m,2H),1.90-2.00(m, 1H),2.15-2.35(m,3H),2.60-2.70(m,1H),3.60-3.66(m,2H),3.97-3.99(m,3H),4 .13-4.15(m,2H),4.30-4.35(m,1H),5.01-5.05(m,2H),5.50-5.59(m,2H),6.30-6 .34(m,3H),6.52-6.60(m,1H),7.16-7.34(m,6H),7.61(s,1H),7.89-7.92(m,1H).

[0667] Compound I-40

[0668] MS m / z[MH] - (ESI): 1121.35; 1 H NMR (300 MHz, methanol-d 4)δ:0.98(s,3H),1.48-1.62(m,4H),1.72-1.88(m,3H),2.12-2.27(m,1H),2.28-2.39(m,2 H),2.46-2.79(m,3H),2.81-2.92(m,2H),2.93-3.03(m,1H),3.14-3.26(m,1H),3.69-3.99 (m,9H),4.21-4.54(m,3H),4.64-4.82(m,3H),4.99-5.09(m,1H),5.38-5.70(m,2H),6.19- 6.36(m,3H),6.43-6.59(m,1H),6.89-7.02(m,1H),7.03-7.16(m,2H),7.17-7.42(m,10H).

[0669] Compound I-41

[0670] MS m / z[MH]-(ESI): 945.30; 1H NMR (400 MHz, methanol-d4) δ: 0.99 (s, 3H), 1.27-1.29 (m, 3H), 1.35-1.36 (m, 3H), 1.56 (s, 4H), 1.78-1.81 (m, 3H), 2.15-2.25 (m, 1H), 2.30-2.34 (m, 2H), 2.50-2.54 (m, 2H), 2.60-2.70 (m, 1H), 2.91-2.99 (m, 2H), 3.81 (s, 3H ),4.21-4.28(m,3H),4.39-4.44(m,1H),4.68-4.77(m,3H),5.04-5.05(m,1H),5.50-5.58(m,2H),6.30-6. 33(m,2H),6.79-6.81(m,1H),6.89-6.94(m,3H),7.11-7.18(m,1H),7.20-7.29(m,2H),7.31-7.39(m,3H).

[0671] Compound I-42

[0672] MS m / z[M / 2-H] - (ESI): 1050.05; 1 H NMR (300 MHz, methanol-d 4)δ: 0.71 - 0.93 (m, 4H), 1.08 (t, J = 7.1 Hz, 4H), 1.17 - 1.31 (m, 1H), 1.47 (s, 4H), 1.67 (s, 3H), 1.76 - 1.95 (m, 2H), 2.00 - 2.17 (m, 2H), 2.19 - 2.33 (m, 4H), 2.32 - 2.55 (m, 6H), 3.04 (t, J = 7.3 Hz, 2H), 3.25 - 3.39 (m, 6H), 3.47 - 3.57 (m, 42H), 3.59 - 3.72 (m, 4H), 3.82 (s, 2H), 4.00 (s, 2H), 4.15 - 4.37 (m, 4H), 4.63 (s, 2H), 4.76 - 5.07 (m, 6H), 5.41 (q, J = 23.0, 21.6 Hz, 3H), 6.24 (d, J = 19.0 Hz, 3H), 6.80 (s, 2H), 6.88 - 7.09 (m, 3H), 7.11 - 7.39 (m, 7H), 8.09 (s, 1H)

[0673] Compound I-43

[0674] MS m / z [M-H] - (ESI): 1105.40; 1 H NMR (300 MHz, methanol-d 4 )δ: 0.88 - 0.98 (m, 6H), 0.99 - 1.02 (m, 1H), 1.03 - 1.19 (m, 1H), 1.27 (s, 1H), 1.39 - 1.44 (m, 2H), 1.47 (s, 3H), 1.51 - 1.64 (m, 4H), 1.66 - 1.73 (m, 1H), 1.81 - 1.89 (m, 3H), 2.02 - 2.33 (m, 4H), 2.31 - 2.39 (m, 1H), 2.46 - 2.53 (m, 2H), 2.58 - 2.69 (m, 3H), 2.91 - 3.09 (m, 3H), 3.11 - 3.19 (m, 1H), 3.21 - 3.29 (m, 1H), 3.71 - 2.91 (m, 6H), 3.92 - 4.11 (m, 2H), 4.39 - 4.41 (m, 1H), 4.48 - 4.58 (m, 2H), 4.61 - 4.66 (m, 1H), 4.69 - 4.79 (m, 2H), 5.99 (s, 1H), 6.12 - 6.28 (m, 1H), 6.76 - 6.88 (m, 1H), 6.89 - 6.99 (m, 3H), 7.11 - 7.31 (m, 6H), 7.32 - 7.44 (m, 1H)

[0675] Compound I-44

[0676] [MH] - (ESI):1193.40. 1 H NMR (300 MHz, methanol-d4) δ 0.98 (s, 3H), 1.57 (s, 4H), 1.64-1.90 (m, 3H), 1.99-2.23 (m, 3H), 2.24-2.4 (m, 2H), 2.42-2.82 (m, 5H), 2.86-3.08 (m, 3H), 3.20 (dd, J = 13.9 Hz, 5.8 Hz, 1H), 3.68-3.95 (m, 6H), 4.02 (t, J = 6.1 Hz, 2H), 4.24-4 .38(m,1H),4.39-4.52(m,2H),4.62(d,J=28.1Hz,1H),4.70-4.81(m,2H),5.04(d,J=4.4Hz,1H),5.37-5.71(m,2 H), 6.16-6.35 (m, 2H), 6.81 (dd, J = 8.0Hz, 1.9Hz, 1H), 6.88-6.98 (m, 3H), 7.03-7.17 (m, 1H), 7.14-7.44 (m, 10H).

[0677] Compound I-45

[0678] MS m / z[MH] - (ESI): 1816.80; 1 H NMR (400 MHz, methanol-d 4)δ:0.79-0.88(m,6H),0.89-0.92(m,1H),0.94-1.04(m,1H),1.13-1.24( m,5H),1.26-1.34(m,3H),1.37(s,3H),1.42-1.51(m,4H),1.58-1.62(m, 1H),1.69-1.71(m,2H),1.91-2.19(m,5H),2.21-2.29(m,1H),2.46-2.61 (m,4H),2.79-2.88(m,15H),2.89-2.99(m,13H),3.11-3.18(m,1H),3.66- 3.81(m,6H),3.91-4.01(m,4H),4.02-4.18(m,7H),4.21-4.24(m,3H),4. 26-4.32(m,5H),4.39-4.48(m,2H),4.49-4.58(m,3H),4.61-4.68(m,3H) ,4.71-4.76(m,2H),5.89(s,1H),5.99-6.22(m,1H),6.61-6.78(m,1H),6 .79-6.98(m,3H),7.01-7.18(m,2H),7.19-7.24(m,4H),7.26-7.38(m,1H)

[0679] Compound I-46

[0680] [MH] - (ESI): 1904.75; 1 H NMR (300 MHz, methanol-d 4 )δ0.98(s,4H),1.29(s,1H),1.56(s,4H),1.79(d,J=13.8Hz,3H),2.01-2.24(m ,4H),2.33(s,2H),2.52-2.78(m,5H),2.81-3.24(m,33H),3.61-3.90(m,6H),3 .90-4.23(m,12H),4.24-4.53(m,11H),4.66-4.81(m,3H),5.03(d,J=4.4Hz,1H ), 5.60 (s, 2H), 6.28 (d, J = 11.7Hz, 2H), 6.74-7.16 (m, 5H), 7.16-7.42 (m, 10H).

[0681] Compound I-47

[0682] MS m / z[M+H] + (ESI): 1019.38

[0683] Compound I-48

[0684] MS m / z[M+H] + (ESI):1728.05; 1 H NMR (400 MHz, methanol-d 4 )δ:0.86-0.90(m,1H),1.02(s,3H),1.22-1.40(m,6H),1.56(s,3H),1.56-1.70(m,1H),1.75- 1.9(m,3H),2.05-2.40(m,3H),2.25-2.45(m,2H),2.50-2.61(m,2H),2.62-2.75(m,2H),2.90 -3.15(m,33H),3.95-4.55(m,24H),4.61-4.82(m,3H),5.05-5.11(m,1H),5.40-5.70(m,2H), 6.25-6.40(m,2H),6.70-6.82(m,1H),6.85-7.00(m,3H),7.05-7.28(m,3H),7.31-7.5(m,3H).

[0685] Compound I-49

[0686] MS m / z[M+H] + (ESI): 2008.9; 1 H NMR (400 MHz, DMSO-d 6)δ9.97 - 9.80 (m, 1H), 8.70 - 8.62 (m, 1H), 8.27 - 8.16 (m, 2H), 7.92 - 7.85 (m, 2H), 7.64 - 7.16 (m, 12H), 7.14 - 7.08 (m, 2H), 6.29 (dd, J=1.6, 10.4 Hz, 1H), 6.12 (s, 1H), 5.72 - 5.47 (m, 2H), 5.05 - 4.89 (m, 3H), 4.86 - 4.57 (m, 3H), 4.38 - 4.24 (m, 2H), 4.19 - 4.17 (m, 2H), 4.12 (t, J=7.6 Hz, 1H), 3.92 - 3.86 (m, 2H), 3.74 - 3.69 (m, 2H), 3.60 - 3.54 (m, 3H), 3.49 - 3.44 (m, 44H), 3.38 (t, J=5.6 Hz, 3H), 3.31 - 3.27 (m, 1H), 3.22 - 3.19 (m, 2H), 3.01 - 2.91 (m, 2H), 2.38 - 2.26 (m, 4H), 2.20 - 2.09 (m, 1H), 1.99 - 1.87 (m, 3H), 1.78 - 1.64 (m, 4H), 1.47 (s, 3H), 1.27 (d, J=7.2 Hz, 3H), 1.06 (t, J=6.8 Hz, 3H), 0.87 - 0.74 (m, 9H).

[0687] Compound I - 50

[0688] MS m / z [M - H] - (ESI): 1670.0; 1 H NMR (400 MHz, DMSO - d 6 )δ8.75 - 8.57 (m, 1H), 8.39 - 7.95 (m, 1H), 7.50 - 7.19 (m, 6H), 7.18 - 6.97 (m, 2H), 6.28 (d, J=10.4 Hz, 1H), 6.12 (s, 1H), 5.75 - 5.50 (m, 2H), 4.93 (d, J=4.0 Hz, 1H), 4.74 - 4.50 (m, 3H), 4.43 - 3.86 (m, 23H), 3.05 - 2.59 (m, 32H), 2.60 - 2.54 (m, 1H), 2.43 (s, 2H), 2.35 - 2.27 (m, 1H), 2.23 - 2.10 (m, 1H), 2.04 - 1.91 (m, 1H), 1.80 - 1.61 (m, 3H), 1.58 - 1.38 (m, 4H), 1.29 - 1.09 (m, 6H), 0.85 (s, 3H)

[0689] Compound I - 51

[0690] MS m / z [M+H] + (ESI): 993.4; 1 H NMR (DMSO-d 6 , 400 MHz) δ 7.73 (d, J = 1.6 Hz, 1H), 7.46 - 7.38 (m, 3H), 7.31 - 7.19 (m, 4H), 7.12 (s, 2H), 6.27 (dd, J = 1.6, 10.0 Hz, 1H), 6.12 (s, 1H), 5.74 - 5.52 (m, 2H), 5.05 (dd, J = 9.6, 18.8 Hz, 1H), 4.94 (d, J = 4.0 Hz, 1H), 4.74 (dd, J = 8.4, 18.8 Hz, 1H), 4.19 (d, J = 8.8 Hz, 1H), 3.91 (q, J = 6.0 Hz, 2H), 3.25 (t, J = 5.6 Hz, 2H), 3.13 (t, J = 8.0 Hz, 2H), 2.45 - 2.38 (m, 2H), 2.31 - 2.26 (m, 1H), 2.20 - 2.12 (m, 1H), 2.02 - 1.92 (m, 1H), 1.82 (d, J = 14.0 Hz, 1H), 1.77 - 1.66 (m, 2H), 1.47 (s, 4H), 0.88 (s, 3H)

[0691] Compound I-52

[0692] MS m / z [M-H] - (ESI): 1701.5896; 1 H NMR (DMSO-d 6 , 400 MHz) δ 7.46 - 7.38 (m, 2H), 7.31 - 7.14 (m, 6H), 7.08 - 7.01 (m, 2H), 6.26 (d, J = 9.6 Hz, 1H), 6.13 (s, 1H), 5.70 - 5.48 (m, 2H), 5.07 - 4.92 (m, 2H), 4.73 (dd, J = 8.8 Hz, 18.0 Hz, 1H), 4.32 - 3.91 (m, 25H), 3.00 - 2.74 (m, 34H), 2.43 (t, J = 7.6 Hz, 2H), 2.25 - 2.15 (m, 1H), 2.04 (d, J = 14.0 Hz, 1H), 1.89 (d, J = 14.0 Hz, 1H), 1.72 (d, J = 8.0 Hz, 2H), 1.50 (s, 4H), 0.92 (s, 3H)

[0693] Compound I-53

[0694] MS m / z [M-H] -(ESI): 1846.1; 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.58 (s, 1H), 8.39 - 8.28 (m, 1H), 8.21 - 8.03 (m, 2H), 8.20 - 8.03 (m, 2H), 7.33 - 7.07 (m, 11H), 6.25 (d, J = 9.6 Hz, 1H), 6.11 (s, 1H), 5.75 - 5.53 (m, 2H), 4.94 (d, J = 4.8 Hz, 1H), 4.72 - 4.56 (m, 3H), 4.50 - 3.85 (m, 22H), 3.78 - 3.65 (m, 6H), 3.11 - 2.68 (m, 30H), 2.54 - 2.52 (m, 1H), 2.27 - 2.25 (m, 1H), 2.23 - 2.10 (m, 2H), 1.99 (d, J = 12.8 Hz, 1H), 1.82 - 1.63 (m, 3H), 1.48 (s, 4H), 0.86 (s, 3H).

[0695] Compound I-54

[0696] MS m / z [M-H] - (ESI): 1678.1; 1 H NMR (400 MHz, DMSO-d 6 ) δ 7.31 (d, J = 10.0 Hz, 1H), 7.22 - 7.13 (m, 1H), 7.05 (s, 2H), 6.85 (d, J = 10.4 Hz, 1H), 6.78 - 6.70 (m, 1H), 6.21 - 6.11 (m, 1H), 6.05 - 5.96 (m, 1H), 5.92 (s, 1H), 5.32 - 5.22 (m, 1H), 4.91 (d, J = 6.8 Hz, 1H), 4.71 (d, J = 3.6 Hz, 1H), 4.65 - 4.40 (m, 3H), 4.38 - 3.96 (m, 24H), 3.01 - 2.66 (m, 36H), 2.45 - 2.26 (m, 4H), 2.09 - 1.90 (m, 6H), 1.82 - 1.64 (m, 6H), 1.61 - 1.47 (m, 3H), 1.35 (s, 3H), 1.27 - 1.11 (m, 7H), 1.05 - 0.88 (m, 2H), 0.79 (s, 3H).

[0697] Compound I-55

[0698] MS m / z [M+Na] + (ESI): 1665.8; 1 H NMR (400 MHz, DMSO-d6 ) δ 8.72 - 8.59 (m, 1H), 7.45 - 7.35 (m, 1H), 7.34 - 7.10 (m, 7H), 7.02 (s, 2H), 6.31 - 6.23 (m, 1H), 6.13 (s, 1H), 5.69 - 5.60 (m, 1H), 5.57 - 5.52 (m, 1H), 4.89 (s, 1H), 4.63 - 4.50 (m, 4H), 4.32 - 4.15 (m, 12H), 2.98 - 2.67 (m, 35H), 2.44 - 2.28 (m, 5H), 2.18 - 2.05 (m, 1H), 1.73 - 1.63 (m, 3H), 1.46 - 1.43 (m, 4H), 0.82 (s, 3H)

[0699] Compound I - 57

[0700] MS m / z [M + H] + (ESI): 1969.9; 1 H NMR (400 MHz, DMSO - d 6 ) δ 8.16 (s, 1H), 7.52 - 6.93 (m, 11H), 6.29 (dd, J = 1.6, 10.0 Hz, 1H), 6.12 (s, 1H), 5.75 - 5.47 (m, 2H), 5.09 - 4.90 (m, 3H), 4.89 - 4.59 (m, 4H), 4.44 - 3.77 (m, 22H), 3.39 - 3.25 (m, 7H), 3.03 - 2.63 (m, 31H), 2.62 - 2.53 (m, 4H), 2.44 - 2.26 (m, 4H), 2.23 - 2.12 (m, 1H), 2.02 - 1.97 (m, 1H), 1.77 - 1.65 (m, 3H), 1.52 - 1.45 (m, 4H), 1.15 - 1.02 (m, 3H), 0.86 (s, 3H)

[0701] Compound I - 58

[0702] MS m / z [M + H] + (ESI): 1672.0; 1 H NMR (400 MHz, DMSO - d 6)δ8.76 - 8.59 (m, 1H), 7.33 - 7.24 (m, 1H), 7.22 - 7.15 (m, 1H), 7.11 (s, 2H), 6.88 (br d, J = 8.8 Hz, 1H), 6.76 (br dd, J = 1.6, 7.6 Hz, 1H), 6.22 (dd, J = 1.6, 10.0 Hz, 1H), 6.01 (s, 1H), 4.74 (br s, 1H), 4.69 - 4.42 (m, 4H), 4.38 - 4.14 (m, 13H), 4.13 - 3.86 (m, 12H), 3.06 - 2.67 (m, 32H), 2.61 (dt, J = 5.2, 13.2 Hz, 1H), 2.47 - 2.37 (m, 3H), 2.36 - 2.21 (m, 2H), 2.07 - 1.64 (m, 13H), 1.55 (br d, J = 8.4 Hz, 2H), 1.46 (s, 3H), 1.36 - 1.25 (m, 1H), 1.25 - 1.19 (m, 3H), 1.19 - 1.12 (m, 3H), 0.81 (s, 3H).

[0703] Compound I - 59

[0704] MS m / z [M + Na] + (ESI): 1675.9; 1 H NMR (400 MHz, DMSO - d 6 )δ8.75 - 8.57 (m, 1H), 7.30 (br d, J = 10.0 Hz, 1H), 7.24 - 7.14 (m, 1H), 7.08 (s, 2H), 6.87 (br d, J = 9.2 Hz, 1H), 6.75 (brdd, J = 1.6, 7.6 Hz, 1H), 6.20 - 6.10 (m, 1H), 5.91 (s, 1H), 4.72 (br d, J = 3.6 Hz, 1H), 4.67 - 4.39 (m, 4H), 4.37 - 4.14 (m, 13H), 4.12 - 3.82 (m, 12H), 3.06 - 2.62 (m, 33H), 2.47 - 2.37 (m, 3H), 2.34 - 2.19 (m, 2H), 2.13 - 1.66 (m, 13H), 1.65 - 1.43 (m, 3H), 1.35 (s, 3H), 1.25 - 1.18 (m, 3H), 1.18 - 1.10 (m, 3H), 1.01 - 0.82 (m, 2H), 0.79 (s, 3H).

[0705] Compound I - 60

[0706] MS m / z [M + Na] + (ESI): 1722.0;1 H NMR (400 MHz, DMSO-d 6 ) δ 8.74 - 8.60 (m, 1H), 7.57 (s, 1H), 7.32 (d, J = 9.6 Hz, 1H), 7.18 (dd, J = 3.2, 10.4 Hz, 1H), 7.12 (s, 2H), 6.97 - 6.86 (m, 2H), 6.80 - 6.71 (m, 1H), 6.21 - 6.14 (m, 1H), 5.96 (d, J = 18.8 Hz, 2H), 5.73 (s, 1H), 4.87 (d, J = 1.6 Hz, 1H), 4.71 - 4.50 (m, 4H), 4.31 (s, 6H), 4.22 (d, J = 14.4 Hz, 9H), 2.94 - 2.75 (m, 35H), 2.43 - 2.38 (m, 3H), 2.34 - 2.29 (m, 2H), 2.06 (s, 5H), 1.98 - 1.88 (m, 3H), 1.82 - 1.68 (m, 4H), 1.65 (s, 3H), 1.38 (s, 4H), 1.24 - 1.11 (m, 9H), 0.85 (s, 4H).

[0707] Compound I-61

[0708] MS m / z [M-H] - (ESI): 1724.1; 1 H NMR (400 MHz, DMSO-d 6 ) δ 7.51 - 7.43 (m, 1H), 7.30 (d, J = 10.0 Hz, 1H), 7.21 - 7.12 (m, 1H), 7.06 (s, 2H), 6.95 (d, J = 6.0 Hz, 1H), 6.90 (d, J = 16.0 Hz, 1H), 6.84 (d, J = 10.0 Hz, 1H), 6.76 - 6.65 (m, 1H), 6.14 (d, J = 9.6 Hz, 1H), 5.91 (s, 1H), 5.87 - 5.76 (m, 1H), 5.12 (d, J = 6.4 Hz, 1H), 4.84 - 4.72 (m, 1H), 4.64 - 4.48 (m, 3H), 4.31 - 4.24 (m, 5H), 4.24 - 4.13 (m, 9H), 4.11 - 3.92 (m, 11H), 2.97 - 2.71 (m, 34H), 2.44 - 2.36 (m, 2H), 2.34 - 2.23 (m, 2H), 2.11 - 1.86 (m, 5H), 1.72 (s, 2H), 1.64 - 1.52 (m, 3H), 1.35 (s, 3H), 1.22 - 1.13 (m, 6H), 1.04 - 0.91 (m, 2H), 0.80 (s, 3H).

[0709] Compound I-62

[0710] MS m / z[MH] - (ESI): 1696.1; 1 H NMR (400 MHz, DMSO-d 6 )δ7.33-7.27(m,1H),7.24-7.17(m,1H),7.10(s,2H),6.88(d,J=9.6Hz,1H),6.83-6.72(m,1H),6.28-6.20( m,1H),6.09-6.00(m,2H),5.27-5.17(m,1H),4.99-4.89(m,1H),4.82-4.74(m,1H),4.67-4.44(m,3H),4.32 (s,4H),4.28-4.15(m,10H),4.14-3.90(m,12H),3.03-2.72(m,34H),2.6-2.58(m,1H),2.45-2.29(m,4H),2 .07-1.89(m,6H),1.88-1.51(m,9H),1.50-1.46(m,3H),1.41-1.29(m,1H),1.27-1.15(m,6H),0.82(s,3H).

[0711] Conjugate

[0712] The compounds of the present disclosure (e.g., compounds of structure (I)) are useful, in part, because they can be linked to a targeting moiety or its targeting fragment (e.g., an antibody). Such a link can be achieved by reducing the disulfide bonds of the protein with an appropriate reagent (e.g., TCEP) and coupling with a compound of structure (I) under appropriate conditions.

[0713] Therefore, another embodiment provides a conjugate having the following structure (II):

[0714]

[0715] in:

[0716] A is a targeting moiety or a binding fragment thereof;

[0717] L 4 Has one of the following structures:

[0718]

[0719] *** indicates the connection point with A;

[0720] g is an integer from 1 to 20;

[0721] X 1 , X 2 , X 3 , X 4 and X 5 One of them is CL 1 -R 1 , X 1 , X 2 , X 3 , X 4 and X 5 The other one is CL 2 -R 2 , and X 1 , X 2 , X 3 , X 4 and X 5 The other three are N, CR, 3 or CL 3 -R 3a ;

[0722] R 1 , R 2 and R 3a Each independently comprises one or more moieties selected from the group consisting of amino acid elements, charged elements, heteroalkylene elements, hydrophilic elements, trigger elements, sacrificial elements, polar caps, payloads, and combinations thereof;

[0723] The condition is R 1 and R 2 At least one of includes a payload;

[0724] R 3 is independently selected at each occurrence from the group consisting of hydrogen, deuterium, alkyl, haloalkyl, halo, alkoxy, haloalkoxy, amino, amine, amido, aldehyde, hydroxyl, cyano, nitro, thiol, carboxyl, carboxyalkyl, alkyl-S(O) 3 H, Alkyl-OP(O) 3 H, Alkyl-P(O) 3 H, -O-carboxyalkyl, -O-alkyl-S(O) 3 H, -O-alkyl-OP(O) 3 H, -O-alkyl-P(O) 3 H, -S(O) 3 H, -OP(O) 3 H, -P(O) 3 H, Alkyl-OP(O) 3 -Alkyl, Alkyl-P(O) 3 -alkyl, -O-alkyl-S(O) 3 -alkyl, -O-alkyl-OP(O)3 -alkyl, -O-alkyl-P(O) 3 -alkyl, -S(O) 3 -alkyl, -OP(O) 3 -Alkyl, -P(O) 3 -Alkyl groups, sulfamides, sulfenamides and carbohydrates;

[0725] R 4a is hydrogen, deuterium, halogenated or -SR 4c , where R 4c is substituted or unsubstituted C 6 -C 10 aryl or substituted or unsubstituted 5-12 membered heteroaryl; and

[0726] L 1 , L 2 and L 3 each is independently a linker comprising an optionally substituted alkylene, an optionally substituted alkenylene, an optionally substituted alkynylene, an optionally substituted heteroalkylene, an optionally substituted heteroalkenylene, an optionally substituted heteroalkynylene, a heteroatom linker, an optionally substituted cycloalkylene, an optionally substituted arylene, an optionally substituted heterocyclylene, an optionally substituted heteroarylene, or a combination thereof, as a stereoisomer, enantiomer or tautomer thereof, or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0727] Another embodiment provides a conjugate having the following structure (II):

[0728]

[0729] in:

[0730] A is a targeting moiety or a binding fragment thereof;

[0731] L 4 Has one of the following structures:

[0732]

[0733] *** indicates the connection point with A;

[0734] g is an integer from 1 to 20;

[0735] X 1 , X 2 , X 3 , X 4 and X 5 One of them is CL 1 -R 1 , X 1 , X 2 , X3 , X 4 and X 5 The other one is CL 2 -R 2 , and X 1 , X 2 , X 3 , X 4 and X 5 The other three are N, CR, 3 or CL 3 -R 3a ;

[0736] R 1 , R 2 and R 3a Each independently comprises one or more moieties selected from the group consisting of amino acid elements, charged elements, heteroalkylene elements, hydrophilic elements, trigger elements, sacrificial elements, polar caps, payloads, and combinations thereof;

[0737] The condition is R 1 and R 2 At least one of includes a payload;

[0738] R 3 is independently selected at each occurrence from the group consisting of hydrogen, deuterium, alkyl, haloalkyl, halo, alkoxy, haloalkoxy, amino, amine, amido, aldehyde, hydroxyl, cyano, nitro, thiol, carboxyl, carboxyalkyl, alkyl-S(O) 3 H, Alkyl-OP(O) 3 H, Alkyl-P(O) 3 H, -O-carboxyalkyl, -O-alkyl-S(O) 3 H, -O-alkyl-OP(O) 3 H, -O-alkyl-P(O) 3 H, -S(O) 3 H, -OP(O) 3 H, -P(O) 3 H, Alkyl-OP(O) 3 -Alkyl, Alkyl-P(O) 3 -alkyl, -O-alkyl-S(O) 3 -alkyl, -O-alkyl-OP(O) 3 -alkyl, -O-alkyl-P(O) 3 -alkyl, -S(O) 3 -alkyl, -OP(O) 3 -Alkyl, -P(O) 3 -Alkyl groups, sulfamides, sulfenamides and carbohydrates;

[0739] R4a is hydrogen, deuterium, halogenated or -SR 4c , where R 4c is substituted or unsubstituted C 6 -C 10 aryl or substituted or unsubstituted 5-12 membered heteroaryl; and

[0740] L 1 , L 2 and L 3 each is independently a direct bond or a linker comprising an optionally substituted alkylene, an optionally substituted alkenylene, an optionally substituted alkynylene, an optionally substituted heteroalkylene, an optionally substituted heteroalkenylene, an optionally substituted heteroalkynylene, a heteroatom linker, an optionally substituted cycloalkylene, an optionally substituted arylene, an optionally substituted heterocyclylene, an optionally substituted heteroarylene, or a combination thereof, as a stereoisomer, enantiomer or tautomer thereof, or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0741] In some embodiments, R 1 Has the following structure:

[0742]

[0743] in:

[0744] L 1a It is an amino acid element;

[0745] L 1b It is a live component;

[0746] L 1c is a heteroalkylene element;

[0747] L 1d It is a hydrophilic element;

[0748] L 1e is a triggering element; and

[0749] L 1f It is a sacrificial component;

[0750] Where L 1a , L 1b , L 1c , L 1d , L 1e and L 1f When present in one or more instances, optionally with L 1a , L 1b , L 1c , L 1d , L 1e and L 1fOne or more of the other of the are joined to form one or more rings;

[0751] n1, n2, n3, n4, n5 and n6 are independently an integer from 0 to 3 at each occurrence, provided that n1+n2+n3+n4+n5+n6=1 or greater;

[0752] n7 is 1, 2, 3, 4, 5 or 6; and

[0753] R 1a Yes and L 1a , L 1b , L 1c , L 1d , L 1e or L 1f A payload is covalently bound in one occurrence and is optionally replaced by a polar cap.

[0754] In certain embodiments, R 1 Has the following structure:

[0755]

[0756] in:

[0757] L 1a It is an amino acid element;

[0758] L 1b It is a live component;

[0759] L 1c is a heteroalkylene element;

[0760] L 1d It is a hydrophilic element;

[0761] L 1e is a triggering element; and

[0762] L 1f It is a sacrificial component;

[0763] Where L 1a , L 1b , L 1c , L 1d , L 1e and L 1f When present in one or more instances, optionally with L 1a , L 1b , L 1c , L 1d , L 1e and L 1f One or more of the other of the are joined to form one or more rings;

[0764] n1, n2, n3, n4, n5 and n6 are independently an integer from 0 to 3 at each occurrence, provided that n1+n2+n3+n4+n5+n6=1 or greater;

[0765] n7 is 1, 2, 3, 4, 5 or 6; and

[0766] R 1a is the payload that is optionally replaced by the polar cap.

[0767] In some embodiments, R 2 Has the following structure:

[0768]

[0769] in:

[0770] L 2a It is an amino acid element;

[0771] L 2b It is a live component;

[0772] L 2c is a heteroalkylene element;

[0773] L 2d It is a hydrophilic element;

[0774] L 2e It is the trigger element;

[0775] m1, m2, m3, m4 and m5 are independently an integer from 0 to 3 at each occurrence, provided that m1+m2+m3+m4+m5=1 or greater;

[0776] m6 is 1, 2, or 3; and

[0777] R 2a is hydrogen, alkyl, payload, or polar cap.

[0778] In certain embodiments, R 3a Has the following structure:

[0779]

[0780] in:

[0781] L 3a It is an amino acid element;

[0782] L 3b It is a live component;

[0783] L 3c is a heteroalkylene element;

[0784] L 3d It is a hydrophilic element;

[0785] L 3e It is the trigger element;

[0786] p1, p2, p3, p4 and p5 are independently an integer from 0 to 3 at each occurrence, provided that p1+p2+p3+p4+p5=1 or greater;

[0787] p6 is 1, 2, or 3; and

[0788] R 3b is hydrogen, alkyl or polar cap.

[0789] In some embodiments, n7 is 1, and each of n1 to n6 is 1. In certain embodiments, n7 is 1, and each of n1 to n4 is 0, n5 is 1, and n6 is 1. In some embodiments, n7 is 1, and n1 is 0, n2 is 0, n3 is 1, n4 is 0, n5 is 1, and n6 is 1.

[0790] In some embodiments, m6 is 1, and each of m1 to m5 is 1. In certain embodiments, m6 is 1, m1 is 1, m2 is 0, m3 is 0, m4 is 1, and m5 is 0.

[0791] In certain embodiments, p6 is 1, and each of p1 to p5 is 1. In some embodiments, p6 is 1, p1 is 1, p2 is 0, p3 is 0, p4 is 1, and p5 is 0.

[0792] In some embodiments, the amino acid element includes one or more amino acids selected from the group consisting of glycine, alanine, serine, threonine, cysteine, valine, leucine, isoleucine, methionine, proline, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, asparagine, glutamine, histidine, lysine, arginine, sarcosine and beta-alanine. In some embodiments, the amino acid element includes one or more amino acids selected from the group consisting of glycine, sarcosine, beta-alanine and glutamic acid.

[0793] In some embodiments, the amino acid element comprises a dipeptide, a tripeptide, a tetrapeptide or a pentapeptide. In some embodiments, the amino acid element has one of the following structures:

[0794]

[0795] in:

[0796] R 5a is independently at each occurrence hydrogen, alkyl, hydroxyalkyl or alkoxyalkyl.

[0797] In some embodiments, the charged element includes a moiety having a negative charge at pH 7.4 (ie, a range of 6.3 to 8.5). In some embodiments, the charged element includes a moiety having a positive charge at pH 7.4 (ie, a range of 6.3 to 8.5).

[0798] In some embodiments, the charged element includes one or more charged amino acids, one or more carboxylic acids, one or more sulfonic acids, one or more sulfonamides, one or more sulfates, one or more phosphates, one or more quaternary amines, one or more sulfamides, one or more sulfenamides, or combinations thereof.

[0799] In some embodiments, the charged amino acid is aspartic acid, glutamic acid, histidine, lysine, or arginine.

[0800] In some embodiments, the heteroalkylene element comprises polyethylene glycol or polypropylene glycol. In some embodiments, the heteroalkylene element comprises one of the following structures:

[0801]

[0802] in:

[0803] R 5b , R 5c , R 5d and R 5e is independently selected at each occurrence from the group consisting of hydrogen, deuterium, alkyl, haloalkyl, halo, alkoxy, haloalkoxy, amino, hydroxy, cyano, nitro, thiol, carboxyalkyl, alkyl-S(O) 3 H, Alkyl-OP(O) 3 H, Alkyl-P(O) 3 H, -O-carboxyalkyl, -O-alkyl-S(O) 3 H, -O-alkyl-OP(O) 3 H, -O-alkyl-P(O) 3 H, -S(O) 3 H, -OP(O) 3 H and -P(O) 3 H,

[0804] q1 is independently an integer from 1 to 24 at each occurrence.

[0805] In some embodiments, q1 is 8, 10, 12 or 14.

[0806] In some embodiments, R 1 , R 2 or R 3a Includes non-cleavable linkers.

[0807] In further embodiments, R 1 , R 2 or R 3a Includes one of the following structures:

[0808]

[0809] in:

[0810] R 5b , R 5c , R 5d and R 5e is independently selected at each occurrence from the group consisting of hydrogen, deuterium, alkyl, haloalkyl, halo, alkoxy, haloalkoxy, amino, hydroxy, cyano, nitro, thiol, carboxyalkyl, alkyl-S(O) 3 H, Alkyl-OP(O) 3 H, Alkyl-P(O) 3 H, -O-carboxyalkyl, -O-alkyl-S(O) 3 H, -O-alkyl-OP(O) 3 H, -O-alkyl-P(O) 3 H, -S(O) 3 H, -OP(O) 3 H, -P(O) 3 H, Alkyl-OP(O) 3 -Alkyl, Alkyl-P(O) 3 -alkyl, -O-alkyl-S(O) 3 -alkyl, -O-alkyl-OP(O) 3 -alkyl, -O-alkyl-P(O) 3 -alkyl, -S(O) 3 -alkyl, -OP(O) 3 -Alkyl, -P(O) 3 -Alkyl, sulfamide, sulfenamide;

[0811] R 5f is independently at each occurrence hydrogen, alkyl, hydroxyalkyl or alkoxyalkyl;

[0812] R 9 is independently at each occurrence hydrogen or alkyl;

[0813] q2 is independently an integer from 1 to 25 at each occurrence; and

[0814] q3 is independently an integer from 5 to 15 at each occurrence.

[0815] In some embodiments, q2 is 8, 10, 12, or 14. In some embodiments, q3 is 6, 8, 10, 12, or 14.

[0816] In some embodiments, the hydrophilic element comprises polyethylene glycol, polysarcosine, cyclodextrin, c-glycoside or a combination thereof. In some embodiments, the hydrophilic element comprises one of the following structures:

[0817]

[0818] in:

[0819] R 5b , R 5c , R 5d and R 5e is independently selected at each occurrence from the group consisting of hydrogen, deuterium, alkyl, haloalkyl, halo, alkoxy, haloalkoxy, amino, hydroxy, cyano, nitro, thiol, carboxyalkyl, alkyl-S(O) 3 H, Alkyl-OP(O) 3 H, Alkyl-P(O) 3 H, -O-carboxyalkyl, -O-alkyl-S(O) 3 H, -O-alkyl-OP(O) 3 H, -O-alkyl-P(O) 3 H, -S(O) 3 H, -OP(O) 3 H and -P(O) 3 H,

[0820] R 5g is independently at each occurrence hydrogen, alkyl, hydroxyalkyl, or alkoxyalkyl; and

[0821] q4 is independently an integer from 1 to 24 at each occurrence.

[0822] In some embodiments, R 5g In each occurrence is –CH 3 .

[0823] In some embodiments, the trigger element comprises a dipeptide, a tripeptide, a tetrapeptide, a pentapeptide, a glucuronide, a disulfide, a phosphate, a diphosphate, a triphosphate, a hydrazone, or a combination thereof. In some embodiments, the trigger element comprises β-glucuronic acid. In certain embodiments, the trigger element comprises a dipeptide, a tripeptide, a tetrapeptide, or a pentapeptide. In certain embodiments, the trigger element comprises two or more amino acids selected from the group consisting of: valine, citrulline, alanine, glycine, phenylalanine, lysine, or a combination thereof. In some embodiments, the trigger element comprises a sequence of amino acids selected from the group consisting of: valine-citrulline, valine-alanine, glycine-glycine-phenylalanine-glycine, and a combination thereof. In certain embodiments, the trigger element comprises a structure in the following structures, including a combination thereof:

[0824]

[0825] In some embodiments, the sacrificial element comprises p-aminobenzyloxycarbonyl, aminal, hydrazine, disulfide, amide, ester, hydrazine, phosphotriester, diester, β-glucuronide, double bond, triple bond, ether bond, ketone, diol, cyano, nitro, quaternary amine, or a combination thereof. In certain embodiments, the sacrificial element comprises p-methoxybenzyl, dialkyldialkoxysilane, diaryldialkoxysilane, orthoester, acetal, optionally substituted β-thiopropionate, ketal, phosphoramidate, hydrazone, vinyl ether, imine, aconityl, trityl, polyketal, bisarylhydrazone, diazobenzene, vicinal diol, pyrophosphate diester, or a combination thereof.

[0826] In some embodiments, the sacrificial element comprises one of the following structures:

[0827]

[0828] In certain embodiments, the sacrificial element comprises the following structure:

[0829]

[0830] in:

[0831] R 6a , R 6b , R 6c and R 6d is independently hydrogen, optionally substituted alkyl, optionally substituted aryl or optionally substituted heteroaryl, or

[0832] R 6a and R 6c together with the nitrogen and carbon atoms to which they are attached form an azetidinyl, pyrrolidinyl, piperidinyl or homopiperidinyl group, and R 6d is hydrogen; and

[0833] Y 1 Is –O–, –S–, or –NR 6b –.

[0834] In some embodiments, the sacrificial element comprises the following structure:

[0835]

[0836] in:

[0837] R 6e , R 6f , R 6g and R 6h is independently hydrogen, optionally substituted alkyl, optionally substituted aryl or optionally substituted heteroaryl, or

[0838] R 6a and R 6c together with the nitrogen and carbon atoms to which they are attached form an azetidinyl, pyrrolidinyl, piperidinyl or homopiperidinyl group, and R 6d is hydrogen; and

[0839] Y 2 Is –O–, –S–, or –NR 6f –.

[0840] In some embodiments, the sacrificial element comprises the following structure:

[0841]

[0842] in:

[0843] R 10 is independently at each occurrence alkyl, alkoxy or halo;

[0844] R 11 is hydrogen, alkyl or –(CH 2 CH 2 O) z3 -CH 3 ;

[0845] R 12 is hydrogen or alkyl;

[0846] R 13 is hydrogen or alkyl;

[0847] z1 is 0 or 1;

[0848] z2 is 0, 1, 2, 3 or 4; and

[0849] z3 is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0850] In some embodiments, the sacrificial element comprises one of the following structures:

[0851]

[0852] in:

[0853] R 14a , R 14b , R 14c , R 14d , R 14e and R 14f each is independently hydrogen, alkyl, hydroxyalkyl or alkoxyalkyl; z4, z5, z6 and z7 are each independently 1, 2, 3, 4, 5 or 6.

[0854] In certain embodiments, the sacrificial element comprises one of the following structures:

[0855]

[0856] in:

[0857] z8 and z9 are each independently 1, 2, 3, 4, 5 or 6; or

[0858] In some embodiments, the sacrificial element comprises one of the following structures:

[0859]

[0860] in:

[0861] R 15 is independently at each occurrence H, methyl, ethyl, isopropyl, tert-butyl or phenyl;

[0862] Y 3 is O or CH 2 ;and

[0863] q5 is an integer in the range of 1-5.

[0864] In some embodiments, R 4a is hydrogen, and is a single bond. In some embodiments, R 4a is hydrogen, and Does not exist.

[0865] In some embodiments, the trigger element and the sacrificial element together include one of the following structures:

[0866]

[0867]

[0868] In some embodiments, the polar cap includes one or more charged amino acids, one or more polyols, or a combination thereof. In certain embodiments, the polar cap includes diols, triols, tetraols, or a combination thereof. In some embodiments, the polar cap includes glycerol, trimethylolpropane, pentaerythritol, maltitol, sorbitol, xylitol, erythritol, isomalt, or a combination thereof. In certain embodiments, the polar cap includes one or more natural amino acids. In some embodiments, the polar cap includes one or more non-natural amino acids. In certain embodiments, the polar cap includes one or more non-natural amino acids and one or more natural amino acids. In some embodiments, the polar cap includes serine, threonine, cysteine, proline, asparagine, glutamine, lysine, arginine, histidine, aspartate, glutamate, 4-hydroxyproline, 5-hydroxylysine, homoserine, homocysteine, ornithine, β-alanine, stadin or γ-aminobutyric acid. In certain embodiments, the polar cap includes aspartic acid, serine, glutamic acid, serine-β-glucose, or a combination thereof.

[0869] In some embodiments, the polar cap has one of the following structures, including combinations thereof:

[0870]

[0871]

[0872] In some embodiments, the payload is a chemotherapeutic agent, a cytotoxic agent, or a myeloid cell agonist.

[0873] In some embodiments, the payload is a chemotherapeutic agent selected from the group consisting of camptothecin, paclitaxel, doxorubicin, vinblastine, dacarbazine, irinotecan, topotecan, xinotecan, cositect, exitecan, lortotecan, SN-38, Dxd, gimatecan, belotecan, and rubitecan.

[0874] In still other embodiments, the payload is a cytotoxic agent selected from the group consisting of calicheamicin, anthramycin, abemycin, cheikamycin, DC-81, methylaminomycin, neoamimycin A, neoamimycin B, puromycin, prolactin, sibamicin, sibirinomycin, tomamycin, auristatin F, monomethyl auristatin F, auristatin E, monomethyl auristatin E, dolestatin, monomethyl dolestatin, maytansine and maytansine.

[0875] In some embodiments, the payload is a myeloid cell agonist selected from a STING agonist, a ligand of TLR2, a ligand of TLR3, a ligand of TLR4, a ligand of TLR5, a ligand of TLR6, a ligand of TLR7, a ligand of TLR8, a ligand of TLR9, a ligand of TLR10, a ligand of a nucleotide oligomerization domain (NOD), a ligand of a RIG-I-like receptor (RLR), a ligand of a C-type lectin receptor (CLR), a cytoplasmic DNA sensor (CD S) and a ligand of an inflammasome inducer, preferably wherein the myeloid cell agonist is selected from the group consisting of sergantomod, motomod, resiquimod, 3M-051, 3M-052, MCT-465, IMO-4200, VTX-763, VTX-1463, RG7854, ADU-S100, MK-1454, MK-2118, BMS-986301, GSK3745417, SB-11285 and IMSA-101.

[0876] In some embodiments, L 1 , L 2 or L 3 A linker comprising a linker selected from the group consisting of: alkylene, alkylene-L a -, alkenylene, alkenylene-L a -, alkynylene, alkynylene-L a -、-L a -、-L a -Alkylene-L a -、-L a -Alkenylene-L a -、-L a -Alkynylidene-L a - and combinations thereof, wherein each alkylene, alkenylene and alkynylene is optionally substituted;

[0877] L a is independently selected at each occurrence from -O-, -S-, -N(R 7 )-, -C(O)-, -C(S)-, -C(O)O-, -OC(O)-, -OC(O)O-, -C(O)N(R 7 )-、-N(R 7 )C(O)-、-C(O)N(R 7 )C(O)-、-C(O)N(R 7 )C(O)N(R 7 )、-N(R 7 )C(O)N(R 7 )-、-N(R 7 )C(O)O-、-OC(O)N(R 7)-、-C(NR 7 )-、-N(R 7 )C(NR 7 )-、-C(NR 7 )N(R 7 )-、-N(R 7 )C(NR 7 )N(R 7 )-、-S(O) 2 -, -OS(O)-, -S(O)O-, -S(O), -OS(O) 2 -、-S(O) 2 O、-N(R 7 )S(O) 2 -、-S(O) 2 N(R 7 )-、-N(R 7 )S(O)-、-S(O)N(R 7 )-、-N(R 7 )S(O) 2 N(R 7 )- and -N(R 7 )S(O)N(R 7 )-;

[0878] R 7 is independently selected at each occurrence from hydrogen, -NH 2 、-C(O)OCH 2 C 6 H 5 ; and C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-12 Cycloalkyl and 3 to 12 membered heterocyclic rings, each of which is optionally substituted with one or more substituents independently selected from the group consisting of halo, hydroxy, cyano, nitro, amino, oxo, thioxo, -C(O)OCH 2 C 6 H 5 、-NHC(O)OCH 2 C 6 H 5 , C 1-10 Alkyl, C 1-10 Haloalkyl, C 1-10 Alkoxy, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-12 Cycloalkyl and 3- to 12-membered heterocyclyl.

[0879] In some embodiments, each L 1 , L 2 or L 3Optionally substituted with one or more substituents selected from the group consisting of alkyl, alkenyl, alkynyl, halo, hydroxy, cyano, -OR 8 、-SR 8 , amino, amine, acylamino, cycloalkyl, aryl, heterocyclyl, heteroaryl, cycloalkylalkyl, arylalkyl, heterocycloalkylalkyl, heteroarylalkyl, -C(O)R 8 、-C(O)N(R 8 ) 2 、-N(R 8 )C(O)R 8 、-C(O)OR 8 、-OC(O)R 8 、-S(O)R 8 、-S(O) 2 R 8 、-P(O)(OR 8 ) 2 、-OP(O)(OR 8 ) 2 , nitro, oxo, thio, =N(R 8 ) or cyano; and

[0880] R 8 is hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heterocyclyl, heteroaryl, cycloalkylalkyl, arylalkyl, heterocycloalkylalkyl or heteroarylalkyl.

[0881] In some embodiments, L 1 , L 2 or L 3 Independently selected from the following structures:

[0882]

[0883] in:

[0884] R a is hydrogen or alkyl;

[0885] L b is independently at each occurrence a direct bond, an optionally substituted alkylene linker, an optionally substituted heteroalkylene linker, a heteroatom linker, or a combination thereof;

[0886] L c is independently at each occurrence an optionally substituted alkylene linker;

[0887] The condition is L 1 , L 2 or L 3 At least one of has the following structure;

[0888]

[0889] In some embodiments, L 1 and L 2 Independently selected from the following structures:

[0890]

[0891] in:

[0892] R a is hydrogen or alkyl;

[0893] L b is independently at each occurrence a direct bond, an optionally substituted alkylene linker, an optionally substituted heteroalkylene linker, a heteroatom linker, or a combination thereof;

[0894] L c is independently at each occurrence an optionally substituted alkylene linker;

[0895] The condition is L 1 or L 2 At least one of has the following structure;

[0896]

[0897] In some embodiments, L 2 Has the following structure:

[0898]

[0899] In some embodiments, L 1 , L 2 and L 3 Each independently has one of the following structures:

[0900]

[0901] in:

[0902] *Indicates a direct bond to a substitutable position on the phenyl group of the following structure:

[0903]

[0904] In some embodiments, L 1 or L 2 Has one of the following structures:

[0905]

[0906] in:

[0907] *Indicates a direct bond to a substitutable position on the phenyl group of the following structure:

[0908]

[0909] In some embodiments, L c It is C 1 -C 6 Alkylene.

[0910] In some embodiments, L c is substituted with one or more substituents selected from the group consisting of halo, haloalkyl, alkoxy, cyano, nitro, carboxyl, sulfonamide, sulfonic acid, or a combination thereof. c is unreplaced.

[0911] In some embodiments, the conjugate has one of the following structures (IIa) or (IIb):

[0912]

[0913] In some embodiments, the conjugate has one of the following structures (IIc') or (IIc"):

[0914]

[0915] In certain embodiments, X 2 , X 3 or both are CH or CF. In some embodiments, X 1 , X 5 or both are CR 3 , and R 3 is H or halo. In some embodiments, X 1 and X 5 All are CR 3 , and R 3 is H or halo. In some embodiments, X 1 Yes CR 3 , and R 3 In certain embodiments, X 5 Yes CR 3 , and R 3 In some embodiments, halo is fluoro. In some embodiments, X 1 In some embodiments, X 1 is CH. In some embodiments, X 5 is CH. In some embodiments, X 5 In some embodiments, X 3 Yes CR 3 , and R 3 is H. In some embodiments, X 3Yes CR 3 , and R 3 is halogenated (eg, fluorinated).

[0916] In some embodiments, the conjugate has one of the following structures: (IIa-1), (IIa-2), (IIa-3), (IIa-4), (IIa-5), or (IIa-6):

[0917]

[0918]

[0919] in:

[0920] L b is independently at each occurrence a direct bond, an optionally substituted alkylene linker, an optionally substituted heteroalkylene linker, a heteroatom linker, or a combination thereof; and

[0921] q6 is 0, 1 or 2.

[0922] In some embodiments, the conjugate has one of the following structures: (IIa-1), (IIa-2), (IIa-3), (IIa-4), (IIa-5), (IIa-6), (IIa-7), or (IIa-8):

[0923]

[0924]

[0925] in:

[0926] L b is independently at each occurrence a direct bond, an optionally substituted alkylene linker, an optionally substituted heteroalkylene linker, a heteroatom linker, or a combination thereof; and

[0927] q6 is 0, 1 or 2.

[0928] In certain embodiments, the conjugate has one of the following structures: (IIc-1), (IIc-2), (IIc-3), or (IIc-4):

[0929]

[0930] in:

[0931] L b is independently at each occurrence a direct bond, an optionally substituted alkylene linker, an optionally substituted heteroalkylene linker, a heteroatom linker, or a combination thereof; and

[0932] q7 is 1, 2 or 3.

[0933] In some embodiments, the conjugate has the following structure (IId), (IIe), (IIf), or (IIg):

[0934]

[0935]

[0936] in:

[0937] L b is independently at each occurrence a direct bond, an optionally substituted alkylene linker, an optionally substituted heteroalkylene linker, a heteroatom linker, or a combination thereof;

[0938] q8 is 0, 1, or 2; and

[0939] q9 is 0, 1, or 2.

[0940] In some embodiments, the conjugate has one of the following structures (IIh) or (IIi):

[0941]

[0942] in:

[0943] L b is independently at each occurrence a direct bond, an optionally substituted alkylene linker, an optionally substituted heteroalkylene linker, a heteroatom linker, or a combination thereof.

[0944] In some embodiments, L b is a direct bond, an optionally substituted alkylene linker, or an optionally substituted heteroalkylene linker.

[0945] In certain embodiments, L b is a direct key or has one of the following structures:

[0946]

[0947] in:

[0948] R b is independently at each occurrence hydrogen, alkyl, hydroxyalkyl or alkoxyalkyl.

[0949] In some embodiments, the conjugate has one of the following structures:

[0950]

[0951]

[0952]

[0953] in:

[0954] R 1b is a chemotherapeutic, cytotoxic, or myeloid cell agonist; R 2b Has one of the following structures:

[0955]

[0956]

[0957] L 1g Has one of the following structures:

[0958]

[0959]

[0960]

[0961] In some embodiments, the conjugate has one of the following structures:

[0962]

[0963]

[0964]

[0965] in:

[0966] R 1b is a chemotherapeutic, cytotoxic, or myeloid cell agonist; R 2b Has one of the following structures:

[0967]

[0968]

[0969] L 1g Has one of the following structures:

[0970]

[0971]

[0972] In certain embodiments, the conjugate has one of the following structures:

[0973]

[0974] In some embodiments, the conjugate has one of the following structures:

[0975]

[0976]

[0977]

[0978] In some embodiments, the conjugate has one of the following structures:

[0979]

[0980]

[0981]

[0982]

[0983]

[0984]

[0985]

[0986] In some embodiments, the conjugate has one of the following structures:

[0987]

[0988]

[0989] In certain embodiments, the conjugate has one of the following structures:

[0990]

[0991]

[0992]

[0993]

[0994]

[0995]

[0996]

[0997]

[0998] In certain embodiments, R1a Has one of the following structures:

[0999]

[1000]

[1001] In some embodiments, R 1a Has one of the following structures:

[1002]

[1003]

[1004] As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[1005] In some embodiments, R 1a Has one of the following structures:

[1006]

[1007]

[1008]

[1009]

[1010] In some embodiments, the payload or R 1a Has one of the following structures:

[1011]

[1012]

[1013] in:

[1014] R' is hydrogen or has one of the following structures:

[1015]

[1016] in:

[1017] R a ' is H or C 1-6 alkyl;

[1018] R b 'It's C 1-6 Alkyl or C 1-6 Alkoxy;

[1019] R c 'It is H, C1-6 Alkyl, -CH 2 OH or C 1-6 Alkoxy;

[1020] R d ' is H or C 1-6 Alkyl; or

[1021] R e ' is H or C 1-6 alkyl.

[1022] In certain embodiments, L 1 or L 2 Has the following structure:

[1023]

[1024] in:

[1025] ** Shown with X 1 , X 2 , X 3 , X 4 or X 5 key.

[1026] In some embodiments, X 2 It is CL 1 -R 1 , and X 3 It is CL 2 -R 2 In some embodiments, X 3 It is CL 1 -R 1 , and X 2 It is CL 2 -R 2 .

[1027] In some embodiments, L 4 Has the following structure:

[1028]

[1029] In some embodiments, L 4 Has the following structure:

[1030]

[1031] In certain embodiments, L 4 Has the following structure:

[1032]

[1033] In some embodiments, the targeting moiety is an anti-CD40 antibody, an antibody selected from the group consisting of an anti-LRRC15 antibody, an anti-CTSK antibody, an anti-ADAM12 antibody, an anti-ITGA11 antibody, an anti-FAP antibody, an anti-NOX4 antibody, an anti-SGCD antibody, an anti-SYNDIG1 antibody, an anti-CDH11 antibody, an anti-PLPP4 antibody, an anti-SLC24A2 antibody, an anti-PDGFRB antibody, an anti-THY1 antibody, an anti-ANTXR1 antibody, an anti-GAS1 antibody, an anti-CALHM5 antibody, an anti-SDC1 antibody, an anti-HER2 antibody, an anti-TROP2 antibody, an anti-MSLN antibody, an anti-fibronectin 4 antibody, an anti-ASGR1 antibody, and an anti-MUC16 antibody.

[1034] In some embodiments, the antibody is a full-length antibody. In certain embodiments, the antibody is an antigen-binding fragment. In some embodiments, the antibody is a humanized antibody. In some embodiments, g is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[1035] Certain embodiments provide a conjugate having one of the structures in Table 2, wherein A is a targeting moiety or a binding fragment thereof, as a stereoisomer, enantiomer or tautomer thereof, or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

[1036]

[1037]

[1038]

[1039]

[1040]

[1041]

[1042]

[1043]

[1044]

[1045]

[1046]

[1047]

[1048]

[1049]

[1050]

[1051]

[1052]

[1053]

[1054]

[1055]

[1056]

[1057]

[1058]

[1059]

[1060]

[1061]

[1062]

[1063]

[1064]

[1065]

[1066]

[1067]

[1068]

[1069]

[1070]

[1071]

[1072]

[1073]

[1074]

[1075]

[1076]

[1077]

[1078]

[1079]

[1080]

[1081] In some embodiments, the targeting moiety is an antibody. The antibody can be of any class, such as IgA, IgD, IgE, IgG and IgM. Several classes in these classes can be further subdivided into isotypes, such as IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2. The heavy chain constant region (Fc) corresponding to different classes of immunoglobulins can be α, δ, ε, γ or μ. Based on the amino acid sequence of the constant domain, the light chain can be one of κ (κ) or λ (λ). The antibody construct can also include an antibody fragment or its recombinant form, including a single chain variable fragment (scFv).

[1082] In certain embodiments, the antibody construct or targeting moiety may include an antigen-binding antibody fragment. The antibody fragment may include (i) a Fab fragment, a monovalent fragment consisting of VL, VH, CL and CH1 domains; (ii) a F(ab')2 fragment, a bivalent fragment comprising two Fab fragments connected by a disulfide bond at the hinge region; and (iii) a Fv fragment consisting of the VL domain and the VH domain of a single arm of an antibody. Although the two domains of the Fv fragment, VL and VH, may be encoded by different genes, they can be connected by a synthetic linker to form a single protein chain in which the VL region and the VH region are paired to form a monovalent molecule.

[1083] In other embodiments, antibody construct or targeting moiety can contain for example two, three, four, five, six, seven, eight, nine, ten or more antigen binding domains.Antibody construct or targeting moiety can contain two antigen binding domains, wherein each antigen binding domain can recognize the same antigen.Antibody construct or targeting moiety can contain two antigen binding domains, wherein each antigen binding domain can recognize different antigens.In some embodiments, antibody construct or targeting moiety can include Fc fusion protein.In other embodiments, antibody construct includes antigen binding domain and Fc region or domain.In still other embodiments, described antibody is chimeric antibody, humanized antibody or people's antibody.

[1084] As used throughout this disclosure, "human antibodies" may include antibodies having, for example, human immunoglobulin amino acid sequences, and antibodies isolated from human immunoglobulin libraries or from one or more human immunoglobulin transgenic animals and that generally do not express endogenous immunoglobulins. Transgenic mice that cannot express functional endogenous immunoglobulins but can express human immunoglobulin genes may be used to produce human antibodies. Fully human antibodies that recognize selected epitopes may be produced using guided selection. In this method, the selected non-human monoclonal antibodies, such as mouse antibodies, are used to guide the selection of fully human antibodies that recognize the same epitope.

[1085] The antibodies or antigen-binding fragments thereof described herein can be derivatized or otherwise modified. For example, the derivatized antibodies can be modified by glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization with known protecting / blocking groups, proteolytic cleavage, etc.

[1086] The antibody, antibody construct or targeting moiety can be a derived antibody. For example, the derived antibody can be modified by glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization with known protection / blocking groups, proteolytic cleavage, connection to a cellular ligand or other protein.

[1087] Antibody constructs may include light chains with at least one, two, three, four, five, six, seven, eight, nine or ten modified amino acid sequences relative to the native or original amino acid sequence, and in some embodiments, no more than 40, 35, 30, 25, 20, 15 or 10 amino acid sequence modifications. Antibody constructs may include heavy chains with at least one, two, three, four, five, six, seven, eight, nine or ten modified amino acid sequences relative to the native or original amino acid sequence, and in some embodiments, no more than 40, 35, 30, 25, 20, 15 or 10 amino acid sequence modifications.

[1088] In any of the embodiments disclosed herein, the antigen binding domain can specifically bind to a tumor antigen. In some embodiments, the tumor antigen is ASGR2, LRRC15, mesothelin (MSLN), HER2, CEA, TROP2, EPHA2, p-cadherin, UPK1B, FOLH1, LYPD3 or PVRL4 (adhesion protein-4). The antigen binding domain can specifically bind to a molecule on an antigen presenting cell (APC). In certain embodiments, the antigen binding domain specifically binds to a human tumor antigen.

[1089] In some embodiments, the antibody (or its antigen binding domain) specifically binds to ASGR1, CTLA4 (also known as CD152), PD-1 (or CD279), PD-L1 (or CD274), TNFR2 (or TNFRSF1B), OX40 (or TNFRSF4), CD27, IL2RA, TNFRSF18, LAG-3, GARP, 4-1BB, ICOS, CD70, PDGFRβ, CD73, CD38, integrin αvβ3, CD248, FAP, integrin αv, or integrin αvβ6.

[1090] In some related embodiments, the targeting moiety is an antibody (e.g., pertuzumab, brentuximab, gemtuzumab, trastuzumab, inotuzumab, polatuzumab, enfortumab, trastuzumab, sacituzumab, belantamab, or moxetumomab).

[1091] In some embodiments, the targeting moiety is an anti-CD40 antibody, an antibody selected from the group consisting of an anti-LRRC15 antibody, an anti-CTSK antibody, an anti-ADAM12 antibody, an anti-ITGA11 antibody, an anti-FAP antibody, an anti-NOX4 antibody, an anti-SGCD antibody, an anti-SYNDIG1 antibody, an anti-CDH11 antibody, an anti-PLPP4 antibody, an anti-SLC24A2 antibody, an anti-PDGFRB antibody, an anti-THY1 antibody, an anti-ANTXR1 antibody, an anti-GAS1 antibody, an anti-CALHM5 antibody, an anti-SDC1 antibody, an anti-HER2 antibody, an anti-TROP2 antibody, an anti-MSLN antibody, an anti-nectin 4 antibody, an anti-ASGR1 antibody, an anti-nectin-4 antibody, and an anti-MUC16 antibody.

[1092] In some embodiments, the targeting moiety is an anti-MSR1 antibody (e.g., as disclosed in PCT Publication No. WO 2019 / 217591, which is incorporated herein by reference).

[1093] Other exemplary antibodies within the scope of the present disclosure will be apparent to those of ordinary skill in the art. For example, exemplary targeting moieties (e.g., antibodies) can be found in PCT Publication Nos. WO 2019 / 217591, WO 2018 / 089373, PCT / US2021 / 054296, or U.S. Patent No. 11,179,473.

[1094] Antibody constructs can be conjugated with joints by cysteine-based bioconjugation. Antibody constructs can be exchanged into appropriate buffers, for example, phosphates, borates, PBS, histidine, triacetates at a concentration of about 2 mg / mL to about 10 mg / mL, and reducing agents of appropriate equivalents, for example, dithiothreitol (DTT) or tris (2-carboxyethyl) phosphine (TCEP). The resulting solution can be stirred for an appropriate amount of time and temperature to achieve the desired reduction. The compound of structure (I) can be added as a solution under stirring. Depending on the physical properties of the compound of structure (I), a cosolvent can be introduced before adding the compound of structure (I) to promote solubility. Depending on the observed reactivity, the reaction can be stirred at room temperature for about 1 hour to about 12 hours. The progress of the reaction can be monitored by liquid chromatography-mass spectrometry (LC-MS). Once it is considered that the reaction is complete, the remaining free structure (I) compound can be removed by an applicable method, and the conjugate of structure (II) can be exchanged into the desired formulation buffer. Such conjugates can be synthesized starting from a targeting moiety (e.g., an antibody or mAb) and a compound of structure (I) (e.g., 7 equivalents) using the conditions described in the conjugation reaction scheme below. The monomer content and drug-antibody ratio can be determined by methods described herein and known in the art.

[1095] Conjugation reaction scheme

[1096]

[1097] Pharmaceutical composition

[1098] The compositions and methods described herein can be considered as pharmaceutical compositions for administration to subjects in need thereof. The pharmaceutical composition can include at least the composition described herein and one or more pharmaceutically acceptable carriers, diluents, excipients, stabilizers, dispersants, suspending agents and / or thickeners. The composition can include a conjugate of structure (II). In some embodiments, the targeting moiety is an anti-LRRC15 antibody. In some embodiments, the targeting moiety is an anti-ASGR1 antibody. The pharmaceutical composition can include a conjugate of structure (II) and one or more of the following: a buffer, an antibiotic, a steroid, a carbohydrate, a drug (e.g., a chemotherapeutic drug), a radioactive substance, a polypeptide, a chelating agent, an adjuvant and / or a preservative.

[1099] One or more physiologically acceptable carriers including excipients and adjuvants can be used to formulate the pharmaceutical composition. The formulation can be adjusted according to the selected route of administration. The pharmaceutical composition comprising the conjugate can be prepared by, for example, lyophilizing the conjugate, mixing, dissolving, emulsifying, encapsulating or embedding the conjugate. The pharmaceutical composition can also contain the conjugate in free base form or in pharmaceutically acceptable salt form.

[1100] The method for preparing the conjugate may include preparing any conjugate with one or more inert, pharmaceutically acceptable excipients or carriers into a solid, semisolid or liquid composition. The solid composition may include, for example, powders, tablets, dispersible granules and capsules, and in some embodiments, the solid composition further contains non-toxic auxiliary substances, such as wetting agents or emulsifiers, pH buffers and other pharmaceutically acceptable additives. Alternatively, the conjugate of structure (II) may be lyophilized or in powder form for reconstitution with a suitable vehicle, such as sterile pyrogen-free water, before use.

[1101] The pharmaceutical composition of the conjugate of structure (II) may include at least one active ingredient (e.g., a compound, salt or conjugate and other agents). The active ingredient may be encapsulated in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization (e.g., hydroxymethylcellulose or gelatin microcapsules and poly(methyl methacrylate) microcapsules, respectively), in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules) or in macroemulsions.

[1102] The pharmaceutical composition may further include more than one active compound as required for the particular indication being treated. The active compounds may have complementary activities and may not adversely affect each other. For example, the composition may further include a chemotherapeutic agent, a cytotoxic agent, a cytokine, a growth inhibitor, an antihormone agent, an antiangiogenic agent and / or a cardioprotectant. Such molecules may be combined in an amount effective for the intended purpose.

[1103] Compositions and formulations may be sterilized. Sterilization may be accomplished by sterile filtration.

[1104] The composition can be formulated as an injection for use. Non-limiting examples of formulations for injection can include sterile suspensions, solutions or emulsions in oily or aqueous vehicles. Suitable oily vehicles can include lipophilic solvents or vehicles, such as fatty oils or synthetic fatty acid esters or liposomes. Aqueous injection suspensions can contain substances that increase the viscosity of the suspension. The suspension can also contain suitable stabilizers. Injections can be formulated for bolus injection or continuous injection. Alternatively, the composition can be lyophilized or in powder form for reconstitution with suitable vehicles, such as sterile pyrogen-free water, before use.

[1105] The phrases "parenteral administration" and "administered parenterally" as used throughout this disclosure mean modes of administration other than enteral and topical administration, usually by injection, and include, but are not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, and intrasternal injection and infusion.

[1106] The phrase "pharmaceutically acceptable" is used herein to refer to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problems or complications commensurate with a reasonable benefit / risk ratio.

[1107] As used throughout this disclosure, the phrase "pharmaceutically acceptable excipient" or "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials that can serve as pharmaceutically acceptable carriers include: (a) sugars, such as lactose, glucose, and sucrose; (b) starches, such as corn starch and potato starch; (c) cellulose and its derivatives, such as sodium carboxymethylcellulose, ethyl cellulose, and cellulose acetate; (d) powdered tragacanth; (e) malt; (f) gelatin; (g) talc; (h) excipients, such as cocoa butter and suppository waxes; (i) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, etc. olive oil, corn oil, and soybean oil; (j) glycols such as propylene glycol; (k) polyols such as glycerol, sorbitol, mannitol, and polyethylene glycol; (l) esters such as ethyl oleate and ethyl laurate; (m) agar; (n) buffers such as magnesium hydroxide and aluminum hydroxide; (o) alginic acid; (p) pyrogen-free water; (q) isotonic saline; (r) Ringer's solution; (s) ethanol; (t) phosphate buffered saline; and (u) other nontoxic compatible substances used in pharmaceutical formulations.

[1108] The term "salt" or "pharmaceutically acceptable salt" refers to salts derived from various organic and inorganic counterions well known in the art. Pharmaceutically acceptable acid addition salts can be formed with inorganic and organic acids. Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like. Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like. Organic bases from which salts can be derived include, for example, primary, secondary and tertiary amines, substituted amines, including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like, specifically such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine and ethanolamine. In some embodiments, the pharmaceutically acceptable base addition salt is selected from ammonium, potassium, sodium, calcium and magnesium salts.

[1109] For parenteral administration, the compound, salt or conjugate can be formulated into an injectable form (e.g., solution, suspension, emulsion) of a unit dose together with a pharmaceutically acceptable parenteral vehicle. Such vehicles may be essentially nontoxic and non-therapeutic. The vehicle can be water, saline, Ringer's solution, dextrose solution, and 5% human serum albumin. Non-aqueous vehicles such as fixed oils and ethyl oleate can also be used. Liposomes can be used as carriers. The vehicle can contain a small amount of additives, such as substances that enhance isotonicity and chemical stability (e.g., buffers and preservatives).

[1110] Sustained release formulations can also be prepared. Examples of sustained release formulations can include semipermeable matrices of solid hydrophobic polymers that can contain compounds, salts, or conjugates, and these matrices can be in the form of molded articles (e.g., films or microcapsules). Examples of sustained release matrices can include polyesters, hydrogels (e.g., poly(2-hydroxyethyl-methacrylate) or poly(vinyl alcohol)), polylactides, copolymers of L-glutamic acid and γ-ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, such as LUPRON DEPO TM (ie, injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate) and other degradable lactic acid-glycolic acid copolymers and poly-D-(-)-3-hydroxybutyric acid.

[1111] A pharmaceutical formulation for storage can be prepared by mixing the conjugate of structure (II) with a pharmaceutically acceptable carrier, excipient and / or stabilizer. This formulation can be a lyophilized formulation or an aqueous solution. Acceptable carriers, excipients and / or stabilizers may be non-toxic to the recipient at the dose and concentration used. Acceptable carriers, excipients and / or stabilizers may include buffers such as phosphates, citrates and other organic acids; antioxidants including ascorbic acid and methionine; preservatives; polypeptides; proteins such as serum albumin or gelatin; hydrophilic polymers; amino acids; monosaccharides, disaccharides and other carbohydrates including glucose, mannose or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counterions such as sodium; metal complexes; and / or nonionic surfactants or polyethylene glycols.

[1112] The pharmaceutical formulation of the conjugate may have an average payload-antibody construct ratio ("DAR") selected from about 1 to about 20, about 1 to about 10, about 1 to about 5, about 1 to about 2, or about 1 to about 1. In some embodiments, the average DAR of the formulation is about 2 to about 8, about 3 to about 8, about 3 to about 7, about 3 to about 5, or about 2. In some embodiments, the average DAR of the pharmaceutical formulation is about 3, about 3.5, about 4, about 4.5, or about 5.

[1113] Treatment

[1114] The conjugates disclosed herein can be used to treat diseases (i.e., conjugates of structure (II)). Those conjugates disclosed herein provide a targeted approach for drug delivery strategies. Therefore, certain embodiments provide a method of treating a disease (or its symptoms), the method comprising administering to a mammal (e.g., a human) in need thereof a therapeutically effective amount of a conjugate of structure (II) or a pharmaceutical composition comprising the conjugate.

[1115] Treat and / or treat refers to any indication of successful treatment or improvement of a disease or condition. Treatment may include, for example, reducing the severity of one or more symptoms of a disease or condition, delaying or alleviating the one or more symptoms, or the treatment may include reducing the frequency with which a patient experiences symptoms of a disease, defect, disorder, or adverse condition. Treatment may be used herein to refer to a method that results in a certain level of treatment or improvement of a disease or condition, and a range of outcomes for the purpose may be considered, including but not limited to preventing the condition entirely.

[1116] Prevent, preventing, and the like refer to preventing a disease or condition, such as tumor formation, in a patient. For example, if an individual at risk for developing a tumor or other form of cancer is treated with the methods of the present disclosure and subsequently does not develop a tumor or other form of cancer, the disease is prevented in the individual, at least for a period of time. Preventing may also refer to preventing the recurrence of a disease or condition in a patient who has previously been treated for the disease or condition, such as by preventing recurrence.

[1117] A therapeutically effective amount (also referred to as an effective amount) can be an amount of a composition comprising a conjugate of structure (II) that is sufficient to provide a beneficial effect or reduce harmful non-beneficial events to an individual to whom the composition is administered. A therapeutically effective dose can be a dose that produces one or more desired or desired (e.g., beneficial) effects after administration, such administration occurring once or multiple times within a given time period. The exact dose can depend on the purpose of the treatment and can be determined by those skilled in the art using known techniques and the teachings provided herein.

[1118] The conjugates that can be used for therapy can be formulated and dosed in a manner consistent with good medical practice, taking into account the disease or condition to be treated, the condition of the individual patient, the site of delivery of the composition, the method of administration, and other factors known to practitioners. The composition can be prepared according to the formulation description described herein.

[1119] The pharmaceutical composition can be used in the methods described herein and can be administered to a subject in need thereof using techniques known to those of ordinary skill in the art, which techniques can be applicable to treating a disease or condition affecting a subject. One of ordinary skill in the art will appreciate that the amount, duration, and frequency of administering the pharmaceutical composition to a subject in need thereof depends on several factors, including, for example, the health of the subject, the specific disease or condition of the patient, the grade or level of the specific disease or condition of the patient, additional treatment that the subject is receiving or has received, etc.

[1120] The conjugates, compositions and methods of the present disclosure can be used as a single agent to treat or prevent diseases such as infection, autoimmune diseases (e.g., multiple sclerosis), inflammation (including autoinflammatory diseases) and cancer. Alternatively, the conjugates, compositions and methods of the present disclosure can be used in combination therapy with a second therapeutic agent for the treatment or prevention of diseases such as infection, autoimmune disorders (e.g., multiple sclerosis), inflammation (including autoinflammatory diseases) and cancer.

[1121] Some embodiments provide a pharmaceutical composition comprising a conjugate of structure (II) and a pharmaceutically acceptable excipient.

[1122] Another embodiment provides a method for treating cancer, comprising administering an effective amount of a conjugate of structure (II) or a pharmaceutical composition thereof to a subject in need thereof. Another embodiment provides a method for treating infection, inflammation, autoimmune disorders, or a combination thereof, comprising administering an effective amount of a conjugate of structure (II) or a pharmaceutical composition thereof to a subject in need thereof. In some of the foregoing embodiments, an effective amount of the conjugate is administered intravenously, subcutaneously, or intratumorally.

[1123] The antigen targeted by the conjugate of structure (II) can be derived from the following specific pathologies and / or families of pathologies, including cancer, such as cancer associated fibroblasts (CAF), brain cancer, skin cancer, lymphoma, sarcoma, lung cancer, liver cancer, leukemia, uterine cancer, breast cancer (including triple negative breast cancer), ovarian cancer, cervical cancer, uterine cancer, bladder cancer, gastric cancer, esophageal cancer, kidney cancer, angiosarcoma, bone cancer, blood cancer, testicular cancer, prostate cancer, gastric cancer, colon cancer, intestinal cancer, pancreatic cancer, head and neck cancer (including head and neck squamous cell carcinoma), mesothelioma, melanoma and other types of cancer as well as precancerous conditions such as hyperplasia, etc.

[1124] Non-limiting examples of additional cancers include acute lymphocytic leukemia (ALL); acute myeloid leukemia (AML); adrenocortical carcinoma; astrocytoma, cerebellum or cerebrum in childhood; basal cell carcinoma; bone tumor, osteosarcoma / malignant fibrous histiocytoma; brain cancer; brain tumors (e.g., cerebellar astrocytoma, malignant glioma, ependymoma, medulloblastoma, visual pathway and hypothalamic glioma); brain stem glioma; breast cancer (including triple negative breast cancer); bronchial adenoma / carcinoid; Burkitt's lymphoma; cerebellar astrocytoma; cervical cancer; bile duct cancer; chondrosarcoma; chronic lymphocytic leukemia; chronic myeloid leukemia; leukemia; chronic myeloproliferative disorder; colon cancer; cutaneous T-cell lymphoma; endometrial cancer; ependymoma; esophageal cancer; eye cancers, such as intraocular melanoma and retinoblastoma; gallbladder cancer; stomach cancer; gliomas; hairy cell leukemia; head and neck cancer (including head and neck squamous cell carcinoma); heart cancer; hepatocellular (liver) cancer; Hodgkin's lymphoma; hypopharyngeal cancer; islet cell carcinoma (endocrine pancreas); Kaposi's sarcoma; kidney cancer (renal cell carcinoma); laryngeal cancer; leukemias, such as acute lymphoblastic, acute myeloid, chronic lymphocytic, chronic myeloid, and hairy cell leukemia; lip and oral cancer; liposarcoma; lung cancers, such as non-small cell Lung cancer and small cell lung cancer; lymphomas, such as AIDS-related Burkitt lymphoma; lymphoma, cutaneous T-cell, Hodgkin and non-Hodgkin, macroglobulinemia, malignant fibrous histiocytoma of bone / osteosarcoma; melanoma (including cutaneous melanoma); Merkel cell carcinoma; mesothelioma; multiple myeloma / plasma cell neoplasms; mycosis fungoides; myelodysplastic syndrome; myelodysplastic / myeloproliferative disorders; myeloproliferative disorders, chronic; nasal and paranasal sinus cancer; nasopharyngeal tumors; neuroblastoma; oligodendroglioma; oropharyngeal cancer; osteosarcoma / malignant fibrous histiocytoma of bone; ovarian cancer; pancreatic cancer; parathyroid cancer; pharyngeal cancer ; pheochromocytoma; pituitary adenoma; plasmacytoma; pleuropulmonary blastoma; prostate cancer; rectal cancer; renal cell carcinoma (kidney cancer); transitional cell carcinoma of the renal pelvis and ureter; rhabdomyosarcoma; salivary gland cancer; sarcoma, Ewing family of tumors; Kaposi's sarcoma; sarcoma, soft tissue; sarcoma, uterine; Sezary syndrome; skin cancer (non-melanoma); skin cancer; small intestine cancer; soft tissue sarcoma; squamous cell carcinoma; primary metastatic squamous neck cancer; gastric cancer; testicular cancer; laryngeal cancer; thymoma and thymic carcinoma; thymoma; thyroid cancer; thyroid cancer, childhood; uterine cancer; vaginal cancer; Waldenstrom's macroglobulinemia; Wilms' tumor, or any combination thereof.

[1125] Additional therapeutic agents that can be combined with the conjugates of the present disclosure can be found in Goodman and Gilman, "The Pharmacological Basis of Therapeutics," Tenth Edition, edited by Hardman, Limbird and Gilman, or the Physician's Desk Reference, both of which are incorporated herein by reference in their entirety.

[1126] The conjugate of structure (II) described herein can be used in combination with the medicament disclosed herein or other suitable medicament, depending on the condition treated. Therefore, in some embodiments, one or more conjugates of the present disclosure will be co-administered with other medicaments as described herein. When used for combination therapy, the conjugate described herein is administered simultaneously or separately with the second medicament. This combination administration may include the simultaneous administration of two medicaments in the same dosage form, the simultaneous administration in a separate dosage form, and the separate administration. That is, the conjugate described herein and any medicament described above can be formulated together in the same dosage form and administered simultaneously. Alternatively, the conjugate of the present disclosure and any of the drugs described above can be administered simultaneously, wherein both medicaments are present in separate formulations. In another alternative, the conjugate of the present disclosure can be administered after any medicament described above, or vice versa. In some embodiments of the single administration scheme, the conjugate of the present disclosure and any medicament described above are administered a few minutes, or hours, or days apart.

[1127] In some embodiments of the present disclosure, including those in which the conjugates and pharmaceutical compositions of the present disclosure are intended for use in treating or preventing inflammation (including autoinflammatory diseases) or autoimmune diseases, the second therapeutic agent comprises an anti-inflammatory composition; a steroid composition, a nonsteroidal anti-inflammatory drug (NSAID) composition, a cyclooxygenase (COX) composition (e.g., a COX1 and / or COX2 inhibitor); or a regulatory T cell antagonist composition.

[1128] In some embodiments of the present disclosure, including those in which the conjugates and pharmaceutical compositions of the present disclosure are intended for the treatment or prevention of cancer, the second therapeutic agent comprises one or more of the following: a second conjugate or pharmaceutical composition of the present disclosure; a chemotherapeutic composition; radiation therapy; an immunoconjugate composition specific for LRRC15, HER2 or MSLN; an immunoconjugate composition specific for an antigen other than LRRC15, HER2 or MSLN; or a modified T cell composition (e.g., CAR-T and / or TCR-T composition).

[1129] In some embodiments, the treatment methods provided herein include administering to the subject an additional therapeutic agent, such as an anticancer agent or an antifibrotic agent. In some embodiments, the additional therapeutic agent is an anticancer agent selected from the group consisting of a chemotherapeutic agent, a cytotoxic agent, a cytokine, a growth inhibitory agent, an antihormonal agent, an antiangiogenic agent, and / or a cardioprotective agent.

[1130] Examples of chemotherapeutic agents contemplated as additional therapeutic agents include alkylating agents, such as nitrogen mustards (e.g., nitrogen mustard, cyclophosphamide, ifosfamide Phenylalanine mustard and chlorambucil); bifunctional chemotherapeutic agents (e.g., bendamustine); nitrosoureas (e.g., carmustine (BCNU, Polyphenylpropanone 20 implant ), lomustine (CCNU), and semustine (methyl-CCNU)); ethyleneimine and methylmelamine (e.g., triethylenemelamine (TEM), triethylenethiophosphoramide (thiotepa), and hexamethylmelamine (HMM, hexamethylmelamine)); alkyl sulfonates (e.g., busulfan Busulfan Injection ); and triazines (e.g., dacarbazine (DTIC)); antimetabolites, such as folic acid analogs (e.g., methotrexate Trimetrexed and pemetrexed (multi-targeted antifolates) and capecitabine Pyrimidine analogs (such as 5-fluorouracil (5-FU, ), fluorodeoxyuridine, tezacitabine, gemcitabine, cytarabine (AraC, cytarabine ); Cytarabine liposome injection ), 5-azacytidine and 2,2'-difluorodeoxycytidine); purine analogs (e.g., 6-mercaptopurine 6-thioguanine, azathioprine, 2'-deoxycoenzyme (pentostatin), erythrohydroxynonyl adenine (EHNA), fludarabine phosphate 2-Chlorodeoxyadenosine (cladribine, 2-CdA); type I topoisomerase inhibitors, such as camptothecin (CPT), topotecan and irinotecan Natural products, such as epipodophyllotoxins (e.g. etoposide and teniposide ); Vinca alkaloids (e.g., vinblastine Vincristine Vinorelbine ); antitumor antibiotics, such as actinomycin D (dacromycin, ), doxorubicin hydrochloride Mitoxantrone and bleomycin sulfate Radiosensitizers, such as 5-bromodeoxyuridine, 5-iododeoxyuridine, and bromodeoxycytidine; platinum coordination complexes, such as cisplatin Carboplatin and oxaliplatin Substituted ureas, such as hydroxyurea Microtubule inhibitors, such as paclitaxel and docetaxel Immunosuppressants, such as cyclophosphamide ( or ); hormone-based compounds, such as anastrozole Exemestane Letrozole Fulvestrant Bicalutamide and tamoxifen citrate Anti-inflammatory agents, such as dexamethasone; anti-androgenic compounds, such as flutamide Anthracyclines, such as idarubicin and epirubicin Bioreductive anticancer agents, such as tirapazamine Serine / threonine kinase inhibitors, such as the CDK4 / 6 inhibitor abemaciclib Pabocini Ribociclib and methylhydrazine derivatives, such as N-methylhydrazine (MIH) and methylbenzylhydrazine.

[1131] Preparation method

[1132] The examples and formulations provided below further illustrate and exemplify the compounds of the present disclosure and methods for preparing such compounds and conjugates thereof. It should be understood that the scope of the present disclosure is not limited in any way by the scope of the following examples and formulations. In the following examples, as well as throughout the specification and claims, unless otherwise indicated, molecules and moieties having a single stereocenter exist as racemic mixtures. Unless otherwise indicated, molecules and moieties having two or more stereocenters exist as racemic mixtures of diastereomers. Single enantiomers / diastereomers can be obtained by methods known to those skilled in the art.

[1133] Those skilled in the art will also appreciate that in the process described herein, the functional groups of the intermediate compounds may need to be protected by suitable blocking groups. Such functional groups include hydroxyl, amino, sulfhydryl and carboxylic acid. Suitable blocking groups for hydroxyl include trialkylsilyl or diarylalkylsilyl (e.g., tert-butyldimethylsilyl, tert-butyldiphenylsilyl or trimethylsilyl), tetrahydropyranyl, benzyl, etc. Suitable blocking groups for amino, amidino and guanidino include tert-butyloxycarbonyl, benzyloxycarbonyl, etc. Suitable blocking groups for sulfhydryl include-C (O) -R "(wherein R " is alkyl, aryl or aralkyl), p-methoxybenzyl, trityl, etc. Suitable blocking groups for carboxylic acid include alkyl, aryl or arylalkyl esters. Blocking groups can be added or removed according to standard techniques known to those skilled in the art and as described herein. The use of protecting groups is described in detail in Green, TW and PGM Wutz, Protective Groups in Organic Synthesis (1999), 3rd edition, Wiley. As will be appreciated by those skilled in the art, the protecting group may also be a polymer resin, such as Wang resin, Rink resin or 2-chlorotrityl-chloride resin.

[1134] In addition, all compounds in the present disclosure that exist in free base or acid form can be converted into their salts by methods known to those skilled in the art by treating with appropriate inorganic or organic bases or acids. Salts of compounds of the present disclosure can be converted into free base or acid forms by standard techniques.

[1135] The following reaction schemes and examples show an exemplary method for preparing compounds of the present disclosure. It should be appreciated that those skilled in the art can prepare these compounds by a similar method or by combining other methods known to those skilled in the art. It should also be appreciated that those skilled in the art can prepare other compounds of structure (I) not specifically described below in a similar manner as described below, by using appropriate starting components and changing synthesis parameters as required. Typically, starting components can be obtained from sources (such as Sigma Aldrich, Lancaster Synthesis, Inc., Maybridge, Matrix Scientific, TCI and Fluorochem, Inc., U.S.), or synthesized according to sources known to those skilled in the art (see, for example, Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 5th edition (Wiley Press, December 2000)), or prepared as described in the present disclosure. The compound 4-((S)-2-((S)-2-amino-3-methylbutanamido)-5-ureidopentanamido)benzyl 3-(2-amino-4-(dipropylcarbamoyl)-3H-benzo[b]azepine-8-carboxamido)-7,8-dihydro-1,6-naphthyridine-6(5H)-carboxylate can be synthesized according to U.S. Pat. No. 10,239,862, the synthesis method of which is hereby incorporated by reference.

[1136] General conjugation protocol

[1137]

[1138] The above general reaction scheme illustrates the conjugation reaction between the compound of structure (I) and a targeting moiety or a binding fragment thereof, wherein mAb-SH represents a targeting moiety or a binding fragment thereof (e.g., a monoclonal antibody) having a free thiol (-SH), and X 1 , X 2 , X 2 , X 4 , X 5 , R 4a and R 4b As defined herein. A targeting moiety or binding fragment thereof having a free thiol can be prepared using methods known in the art and as described herein (see, e.g., Conjugation Example 1) and conjugated to a compound of structure (I). After the initial conjugation reaction, the resulting conjugate of structure (II) can undergo an irreversible hydrolysis reaction to form an alternative conjugate of structure (II) as shown.

[1139] Reaction Scheme 1

[1140]

[1141]

[1142]

[1143]

[1144] Intermediate Example 1

[1145] Preparation of tert-butyl 2-[[2-(benzyloxycarbonylamino)acetyl]amino]acetate

[1146]

[1147] To a solution of 2-(benzyloxycarbonylamino)acetic acid (5.50 g, 26.3 mmol, 1.0 eq.) in DMF (100 mL) were added triethylamine (TEA; 7.98 g, 78.9 mmol, 11.0 mL, 3.0 eq.), HATU (12.0 g, 31.5 mmol, 1.2 eq.) and tert-butyl 2-aminoacetate (3.45 g, 26.3 mmol, 1.0 eq.), and then stirred at 25° C. for 2 hours. The reaction mixture was stirred at 0° C. by adding H 2 O (150 mL), and then extracted with ethyl acetate (100 mL x 3). The combined organic layers were washed with H 2 O (80 mL × 3), the fuel oil was washed with brine (100 mL), and then 2 SO 4 The residue was dried, filtered and concentrated under reduced pressure to give a residue. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, ethyl acetate / methanol=1 / 1) to give tert-butyl 2-[[2-(benzyloxycarbonylamino)acetyl]amino]acetate (8 g, 24.82 mmol, 94.40% yield) obtained as a yellow oil.

[1148] 1 H NMR (CDCl 3 ,400MHz)δ7.40-7.31(m,5H),5.14(s,2H),3.98-3.89(m,4H),1.47(s,9H)

[1149] Intermediate Example 2

[1150] Preparation of tert-butyl 2-[(2-aminoacetyl)amino]acetate

[1151]

[1152] In N 2 To a solution of tert-butyl 2-[[2-(benzyloxycarbonylamino)acetyl]amino]acetate (8.80 g, 27.3 mmol, 1.0 equiv) in methanol (150 mL) was added Pd / C (10%, 2.5 g) under atmospheric pressure. The suspension was degassed and heated with H 2 Purge 3 times. The mixture was stirred under H 2 The mixture was stirred at 25° C. for 2 hours under reduced pressure (50 Psi). The reaction mixture was filtered and concentrated under reduced pressure to give a residue. Compound tert-butyl 2-[(2-aminoacetyl)amino]acetate (4.2 g, 22.31 mmol, 81.74% yield) was obtained as a white solid.

[1153] 1 H NMR (CDCl 3 ,400MHz)δ7.67(s,1H),3.96(d,J=5.6Hz,2H),3.39(s,2H),1.75(s,2H),1.46(s,9H)

[1154] Intermediate Example 3

[1155] Preparation of (E)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)prop-2-enoic acid methyl ester

[1156]

[1157] To a solution of CuCl (1.06 g, 10.7 mmol, 256 μL, 0.03 eq.) and Xantphos (6.19 g, 10.7 mmol, 0.03 eq.) in THF (450 mL) was added t-BuONa (2.06 g, 21.4 mmol, 0.06 eq.) at 0°C. The reaction solution was stirred at 0°C for 1 hour, and then Pin was added. 2 B 2(90.6g, 357mmol, 1.0 equivalent) in THF (150mL). The reaction solution was stirred at 20°C for one hour under a nitrogen atmosphere. To the above reaction solution, methyl prop-2-ynoate (30.0g, 357mmol, 29.7mL, 1.0 equivalent) and methanol (22.9g, 714mmol, 28.9mL, 2.0 equivalent) were added. And the reaction solution was stirred at 20°C for another 12 hours. The resulting mixture was poured into ice water (w / w=1 / 1) (300mL) and stirred for 5 minutes. The aqueous phase was extracted with ethyl acetate (200mL×3). The combined organic phase was washed with brine (50mL×2) and washed with anhydrous Na 2 SO 4 The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether / ethyl acetate = 1 / 0, 3 / 1) to give (E)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)prop-2-enoic acid methyl ester (70 g, 330 mmol, 92.51% yield) as a colorless oil.

[1158] 1 H NMR (400 MHz, CDCl 3 )δ6.83-6.75(m,1H),6.68-6.59(m,1H),3.77(s,3H),1.29(s,12H).

[1159] Intermediate Example 4

[1160] Preparation of tert-butyl 2-bromo-5-nitrobenzoate

[1161]

[1162] At 0 ° C, to a mixture of 2-bromo-5-nitrobenzoic acid (9.70 g, 39.4 mmol, 1.0 equiv) in DCM (100 mL), DCC (8.95 g, 43.4 mmol, 8.77 mL, 1.1 equiv) and DMAP (2.41 g, 19.7 mmol, 0.5 equiv) were added. The mixture was stirred at 0 ° C for 10 minutes, then t-BuOH (4.38 g, 59.1 mmol, 5.66 mL, 1.5 equiv) was added and stirred at 20 ° C for 12 hours. The mixture was poured into ice water (w / w=1 / 1) (50 mL) and stirred for 10 minutes. The aqueous phase was extracted with DCM (50 mL×2). The combined organic phase was washed with brine (10 mL×2) and washed with anhydrous Na 2 SO 4Dry, filter and concentrate under vacuum. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether / ethyl acetate=1 / 0, 5 / 1) to give tert-butyl 2-bromo-5-nitrobenzoate (9.2 g, 30.5 mmol, 77.23% yield) as a white solid.

[1163] 1 H NMR (400 MHz, CDCl 3 )δ8.52(d,J=2.8Hz,1H),8.13(dd,J=2.8,8.8Hz,1H),7.83(d,J=8.8Hz,1H),1.65(s,9H).

[1164] Intermediate Example 5

[1165] Preparation of tert-butyl 2-[(E)-3-methoxy-3-oxo-1-enyl]-5-nitro-benzoate

[1166]

[1167] (E)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)prop-2-enoic acid methyl ester (11.2 g, 53.0 mmol, 2.5 equivalents), tert-butyl 2-bromo-5-nitrobenzoate (6.40 g, 21.2 mmol, 1.0 equivalents), K 3 PO 4 (6.74 g, 31.8 mmol, 1.5 eq.), s-Phos (870 mg, 2.12 mmol, 0.1 eq.), Pd 2 (dba) 3 (970 mg, 1.06 mmol, 0.05 equiv) in dioxane (120 mL) and H 2 The mixture was degassed with N 2 Purge 3 times, and then at N 2 The mixture was stirred at 90°C for 12 hours under a 30% atmosphere. The mixture was poured into ice water (w / w=1 / 1) (30 mL) and stirred for 10 minutes. The aqueous phase was extracted with ethyl acetate (50 mL×3). The combined organic phases were washed with brine (10 mL×3) and washed with anhydrous Na 2 SO 4The residue was dried, filtered and concentrated under vacuum. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether / ethyl acetate=1 / 0, 3 / 1) to give 2-[(E)-3-methoxy-3-oxo-prop-1-enyl]-5-nitro-benzoic acid tert-butyl ester (6 g, 19.5 mmol, 92.17% yield) as a yellow solid.

[1168] 1 H NMR (400 MHz, CDCl 3 )δ8.74(d,J=2.4Hz,1H),8.41(d,J=15.6Hz,1H),8.34(dd,J=2.4,8.4Hz,1H ), 7.72 (d, J = 8.4Hz, 1H), 6.38 (d, J = 15.6Hz, 1H), 3.85 (s, 3H), 1.65 (s, 9H).

[1169] Intermediate Example 6

[1170] Preparation of tert-butyl 5-amino-2-(3-methoxy-3-oxo-propyl)benzoate

[1171]

[1172] A mixture of 2-[(E)-3-methoxy-3-oxo-prop-1-enyl]-5-nitro-benzoic acid tert-butyl ester (4.00 g, 13.0 mmol, 1.0 equiv), Pd / C (400 mg, 13.0 mmol, 10% purity, 1.0 equiv) in methanol (50 mL) was degassed and heated with H 2 (26.2 mg, 13.0 mmol) was purged 3 times, and the mixture was then stirred under H 2 Stir under atmosphere at 25° C. for 4 hours. The mixture was filtered and concentrated in vacuo to give tert-butyl 5-amino-2-(3-methoxy-3-oxo-propyl)benzoate (3.50 g, 12.5 mmol, 96.26% yield) as a yellow oil.

[1173] 1 H NMR (400 MHz, CDCl 3 )δ7.14(d,J=2.4Hz,1H),7.04(d,J=8.0Hz,1H),6.73(dd,J=2.4,8.0Hz,1H),3.67(s,3H),3.12(t,J=8.0Hz,2H),2.61(t,J=8.0Hz,2H),1.59(s,9H).

[1174] Intermediate Example 7

[1175] Preparation of 5-amino-2-(3-methoxy-3-oxopropyl)benzoic acid

[1176]

[1177] To a solution of tert-butyl 5-amino-2-(3-methoxy-3-oxo-propyl)benzoate (3.50 g, 12.5 mmol, 1.0 equiv) in ethyl acetate (10 mL) was added HCl / ethyl acetate (4 M, 50 mL, 16.0 equiv) and then stirred at 25° C. for 12 hours. The mixture was concentrated in vacuo to give 5-amino-2-(3-methoxy-3-oxo-propyl)benzoic acid (3.20 g, 12.3 mmol, 98.35% yield, HCl) as a white solid.

[1178] 1 H NMR (400 MHz, DMSO-d 6 )δ7.69(d,J=2.0Hz,1H),7.39-7.31(m,2H),3.57(s,3H),3.15(t,J=7.6Hz,2H),2.59(t,J=7.6Hz,2H).

[1179] Intermediate Example 8

[1180] Preparation of methyl 3-[4-amino-2-[[2-[(2-tert-butoxy-2-oxo-ethyl)amino]-2-oxo-ethyl]carbamoyl]phenyl]propanoate

[1181]

[1182] At 0 ° C, to a solution of 5-amino-2-(3-methoxy-3-oxo-propyl)benzoic acid (3.20 g, 12.3 mmol, 1.0 eq., HCl) in DMF (40 mL), NMM (3.74 g, 37.0 mmol, 4.06 mL, 3.0 eq.), HOBt (833 mg, 6.16 mmol, 0.5 eq.), EDCI (4.72 g, 24.7 mmol, 2.0 eq.) and tert-butyl 2-[(2-aminoacetyl)amino]acetate (2.78 g, 14.8 mmol, 1.2 eq.) were added, and then stirred at 20 ° C. for 1 hour. The mixture was poured into ice water (w / w=1 / 1) (40 mL) and stirred for 10 minutes. The aqueous phase was extracted with ethyl acetate (50 mL×3). The combined organic phase was washed with brine (20 mL×2) and washed with anhydrous Na 2 SO 4The residue was dried, filtered and concentrated under vacuum. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether / ethyl acetate=1 / 0, 0 / 1) to give methyl 3-[4-amino-2-[[2-[(2-tert-butoxy-2-oxo-ethyl)amino]-2-oxo-ethyl]carbamoyl]phenyl]propanoate (3.8 g, 9.66 mmol, 78.38% yield) as a yellow oil.

[1183] 1 H NMR (400 MHz, CDCl 3 )δ7.04(d,J=8.0Hz,2H),6.79-6.66(m,3H),4.15(d,J=5.6Hz,2H),3.97(d,J=5. 2Hz, 2H), 3.61 (s, 3H), 2.99 (t, J = 7.2Hz, 2H), 2.68 (t, J = 7.2Hz, 2H), 1.47 (s, 9H).

[1184] Intermediate Example 9

[1185] Preparation of 2-[[2-[[5-amino-2-(3-methoxy-3-oxo-propyl)benzoyl]amino]acetyl]amino]acetic acid

[1186]

[1187] To a solution of methyl 3-[4-amino-2-[[2-[(2-tert-butoxy-2-oxo-ethyl)amino]-2-oxo-ethyl]carbamoyl]phenyl]propanoate (0.60 g, 1.53 mmol, 1.0 equiv) in ethyl acetate (5 mL) was added HCl / ethyl acetate (4 M, 10 mL, 26.2 equiv) and then stirred at 20° C. for 12 hours. The mixture was concentrated in vacuo to give 2-[[2-[[5-amino-2-(3-methoxy-3-oxo-propyl)benzoyl]amino]acetyl]amino]acetic acid (550 mg, 1.47 mmol, 96.48% yield, HCl) as a white solid.

[1188] 1 H NMR (400 MHz, DMSO-d 6)δ8.57(t,J=6.0Hz,1H),8.25(t,J=6.4Hz,1H),7.28(d,J=8.0Hz,1H),7.20-7.10(m,2H),3.89(d ,J=6.0Hz,2H),3.81(d,J=6.0Hz,2H),3.57(s,3H),2.90(t,J=8.0Hz,2H),2.60(t,J=8.0Hz,2H).

[1189] Intermediate Example 10

[1190] Preparation of intermediate 1a

[1191]

[1192] 2-[[2-[[5-amino-2-(3-methoxy-3-oxo-propyl)benzoyl]amino]acetyl]amino]acetic acid (320 mg, 856 μmol, 1.0 equiv, HCl), 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2-aminoethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy] A mixture of tert-butyl]ethoxy]ethoxy]propanoate (576 mg, 856 μmol, 1.0 equiv), EDCI (328 mg, 1.71 mmol, 2.0 equiv), HOBt (57.8 mg, 428 μmol, 0.5 equiv) and 4-methylmorpholine (259 mg, 2.57 mmol, 282 μL, 3.0 equiv) in DMF (5 mL) was stirred at 25 °C for 2 h. The reaction mixture was heated at 0 °C by adding H 2 O (10 mL) quenched, and then extracted with DCM / i-PrOH (v: v = 3: 1, 10 mL x 5). The combined organic layer was concentrated under reduced pressure to give a residue. The desired product (950 mg, crude product) was obtained as a yellow oil. [M + H] + (ESI):994.10.

[1193] Intermediate Example 11

[1194] Preparation of intermediate 2a

[1195]

[1196] To a solution of intermediate 1a (950 mg, 956 μmol, 1.0 equiv) in ethyl acetate (5 mL) was added HCl / ethyl acetate (15 mL, 4 M). The mixture was stirred at 25 °C for 2 hours. The reaction mixture was concentrated under reduced pressure. The desired product (890 mg, crude product) was obtained as a yellow solid.

[1197] In another reaction, hydrogen chloride (gas) / ethyl acetate (15 mL, 4 mol / L) was added to a solution of intermediate 1a (850 mg, 956 μmol, 1.0 equiv) in dioxane (5.0 mL). The mixture was stirred at 25 °C for 2 hours. The reaction mixture was concentrated under reduced pressure. Intermediate 2a (800 mg, crude product) was obtained as a yellow solid. [M+H] + (ESI):936.20.

[1198] Intermediate Example 12

[1199] Preparation of intermediate 3a

[1200]

[1201] A mixture of Intermediate 2 (880 mg, 903 μmol, 1.0 eq., HCl), di-tert-butyl (2S)-2-aminopentanedioate (588 mg, 1.99 mmol, 2.2 eq., HCl), EDCI (346 mg, 1.81 mmol, 2.0 eq.), HOBt (61.0 mg, 451 μmol, 0.5 eq.) and 4-methylmorpholine (274 mg, 2.71 mmol, 298 μL, 3.0 eq.) in DMF (10 mL) was stirred at 25 °C for 2 h. The reaction mixture was heated at 0 °C by adding H 2 O (20 mL) quenched, and then extracted with DCM / i-PrOH (v:v = 3: 1, 20 mL x 5). The combined organic layer was concentrated under reduced pressure to give a residue. The desired product (900 mg, 763 μmol, 84.49% yield) was obtained as a yellow oil.

[1202] In another reaction, a mixture of intermediate 2a (800 mg, 0.903 mmol, 1.0 equiv), (2S)-2-aminopentanedioic acid di-tert-butyl ester (588 mg, 1.99 mmol, 2.2 equiv, HCl), 1-ethyl-3 (3-dimethylpropylamine) carbodiimide (346 mg, 1.81 mmol, 2.0 equiv), 1-hydroxybenzotriazole (61.0 mg, 0.451 mmol, 0.5 equiv) and 4-methylmorpholine (274 mg, 2.71 mmol, 3.0 equiv) in N, N-dimethylformamide (10.0 mL) was stirred at 25 ° C for 2 hours. The reaction mixture was quenched at 0 ° C by adding water (20 mL), and then extracted with dichloromethane / isopropanol (v:v=3:1, 50 mL×5). The combined organic layer was concentrated under reduced pressure to give a residue. Column, C 18Silica gel; mobile phase, acetonitrile in water (0.1% trifluoroacetic acid / formic acid), gradient 10% to 70% in 20 minutes; detector, UV 254 nm. 900 mg (89%) of intermediate 3a were obtained as a yellow oil. [M+H] + (ESI): 1179.05

[1203] Intermediate Example 13

[1204] Preparation of intermediate 4a

[1205]

[1206] Intermediate 3a (800 mg, 679 μmol, 1.0 eq.) was dissolved in methanol (2.5 mL) and H 2 To a solution in 4% 4-nitropropene (2.5 mL) was added NaOH (136 mg, 3.39 mmol, 5.0 equiv), and then stirred at 20° C. for 1 hour. The mixture was concentrated in vacuo to remove methanol, and then washed with H 2 O (20 mL), the pH of the aqueous phase was adjusted to about 7 with HCl (2 M), and extracted with DCM / i-PrOH (v:v=3:1, 15 mL×5). The combined organic layers were purified by Na 2 SO 4 Dry, filter and concentrate under reduced pressure to give a residue to afford the desired product (700 mg, crude) as a red oil.

[1207] In another reaction, sodium hydroxide (136 mg, 3.39 mmol, 5.0 equiv) was added to a solution of intermediate 3a (800 mg, 0.68 mmol, 1.0 equiv) in methanol (8.0 mL) and water (2.5 mL), and then stirred at 0 ° C for 2 hours. The mixture was concentrated in vacuo to remove methanol, and then diluted with water (20 mL), the pH of the aqueous phase was adjusted to about 7 with hydrochloric acid (2 mol / L), and extracted with dichloromethane / isopropanol (v / v = 3: 1, 30 mL × 5). The combined organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure to give a residue to give intermediate 4a (700 mg, crude product) as a red oil. [M + H] + (ESI):1165.10.

[1208] Intermediate Example 14

[1209] Preparation of intermediate 5a

[1210]

[1211] To a solution of intermediate 4a (250 mg, 214 μmol, 1.0 eq) and methyl 2,5-dioxopyrrole-1-carboxylate (49.9 mg, 322 μmol, 49.9 μL, 1.5 eq) in DCM (3 mL) was added TEA (65.1 mg, 644 μmol, 89.6 μL, 3.0 eq) and then stirred at 50° C. for 4 h. The pH of the mixture was adjusted to about 6 with TFA and quenched by adding H 2 The reaction mixture was diluted with 4% paraformaldehyde (2% paraformaldehyde) and 4% paraformaldehyde (1% paraformaldehyde) (20% paraformaldehyde, 1 ...

[1212] In another reaction, trimethylamine (182.2 mg, 1.80 mmol, 3.0 equiv) was added to a solution of intermediate 4a (700 mg, 0.60 mmol, 1.0 equiv) and methyl 2,5-dioxopyrrole-1-carboxylate (139.8 mg, 0.92 mmol, 1.5 equiv) in dichloromethane (7.0 mL), and then stirred at 50 ° C for 4 hours. The pH of the mixture was adjusted to about 6 with trifluoroacetic acid, and diluted by adding water (10 mL), and extracted with dichloromethane: isopropanol (v / v=3:1, 30 mL×5). The combined organic layers were concentrated under reduced pressure to give a residue. Column, C 18 Silica gel; mobile phase, acetonitrile in water (0.1% trifluoroacetic acid / formic acid), gradient 10% to 70% in 20 minutes; detector, UV 254 nm. 306 mg (41%) of intermediate 5a was obtained as a colorless oil. [M+H] + (ESI):1245.32.

[1213] Intermediate Example 15

[1214] Preparation of intermediate 6a

[1215]

[1216] A mixture of intermediate 5a (270 mg, 216 μmol, 1.0 equiv) and EDCI (166 mg, 867 μmol, 4.0 equiv) in DCM (2 mL) and DMA (2 mL) was stirred at 25 ° C for 2 hours. The reaction mixture was concentrated under reduced pressure to remove DCM. The residue was purified by preparative HPLC (column: Phenomenex Luna 80 30 mm 3 μm; mobile phase: [water (TFA)-acetonitrile]; B%: 35%-70%, 8 minutes). The eluent was removed under freeze drying. The desired product (90 mg, 64.6 μmol, 29.79% yield) was obtained as a colorless oil.

[1217] 1 H NMR (DMSO-d 6 ,400MHz)δ7.94-7.84(m,1H),7.50(d,J=8.4Hz,1H),7.43-7.34(m,2H),7.18(s,2 H),4.13(dd,J=5.2,9.1Hz,1H),3.89(s,2H),3.70(s,2H),3.60-3.58(m,3H),3.5 1-3.47(m,44H),3.39(t,J=6.0Hz,2H),3.22-3.17(m,2H),3.14(s,3H),2.42-2.3 2(m,2H),2.27-2.19(m,2H),1.95-1.83(m,1H),1.77-1.65(m,1H),1.38(s,18H).

[1218] In another reaction, a mixture of 2,3,5,6-tetrafluorophenol (108 mg, 0.65 mmol, 3.0 equiv), intermediate 5a (270 mg, 216 μmol, 1.0 equiv) and 1-ethyl-3 (3-dimethylpropylamine) carbodiimide (166 mg, 867 μmol, 4.0 equiv) in N,N-dimethylformamide (3.0 mL) was stirred at 25 ° C for 2 hours. The reaction mixture was concentrated under reduced pressure. The crude product was purified by flash preparative HPLC under the following conditions (IntelFlash-1): column, C 18 Silica gel; mobile phase, water and acetonitrile (10.0% acetonitrile increased to 100.0% in 20 minutes); detector, UV 254nm. 90 mg (30%) of intermediate 6a was obtained as a colorless oil. [M+H] + (ESI):1394.45.

[1219] Alternatively, intermediate 6a can be deprotected according to the following reaction scheme and protocol:

[1220]

[1221] At 0°C, under an inert nitrogen atmosphere, trifluoroacetic acid (0.25 mL) was added to a solution of intermediate 6a (90 mg, 0.07 mmol, 1.0 equiv) in dichloromethane (1.0 mL). The resulting solution was stirred at 0°C for 4 hours and concentrated in vacuo and under reduced pressure. 90 mg (crude product) of intermediate 6a' was obtained as a yellow oil. [M+H] + (ESI):1279.95.

[1222] Intermediate Example 16

[1223] Preparation of intermediate 7a

[1224]

[1225] To a solution of 3-[[2-amino-4-(dipropylcarbamoyl)-3H-1-benzazepine-8-carbonyl]amino]-7,8-dihydro-5H-1,6-naphthyridine-6-carboxylic acid [4-[[(2S)-2-[[(2S)-2-amino-3-methyl-butanoyl]amino]-5-ureido-pentanoyl]amino]phenyl]methyl ester (20.0 mg, 23.0 μmol, 1.0 equiv) in DMF (0.5 mL) was added diisopropylethylamine (DIEA) (8.95 mg, 69.2 μmol, 12.0 μL, 3.0 equiv) and intermediate 6a (32.1 mg, 23.0 μmol, 1.0 equiv). The mixture was stirred at 25 °C for 1 hour. At 0 °C, the pH of the mixture was adjusted to about 6 with trifluoroacetic acid (TFA) and the reaction mixture was heated to 40 °C by adding H 2 The reaction mixture was diluted with 4% paraformaldehyde (5% ethanol) and extracted with dichloromethane (DCM):isopropyl alcohol (i-PrOH) (v:v=3:1, 5 mL×5). The combined organic layers were concentrated under reduced pressure to give a residue. The desired product (55 mg, crude product) was obtained as a yellow oil.

[1226] Intermediate Example 17

[1227] Synthesis of (19S,22R,23R,23AR,25R,27AR,29R,210R,210AR,212S,214AR,37R,E)-23,210-difluoro-25,212-dimercapto-23,23A,27A,29,210,210A,214,214A-octahydro-19H,22H,27H,37H-4,9-diaza-1(9,6)-purine-3(7,4)-pyrrolo[2,3-D]pyrimidine-2(2,9)-difluoro[3,2-D:3',2'-J][1,3,7,9]tetraoxa[2,8]diphosphocyclododecanoylnonane-6-ene 25,212-dioxide

[1228]

[1229]

[1230]

[1231]

[1232]

[1233]

[1234] Intermediate 1b (alternatively its ammonium salt form) is obtained using the reaction conditions and synthesis scheme shown above. Reaction methods, reagents, purification techniques, etc. are known in the art. For example, certain aspects of the method for preparing the compounds of the present disclosure can be found in PCT Publication Nos. 2018 / 152453 and / or 2018 / 152450, which are hereby incorporated by reference in their entirety.

[1235] (2R,3R,4R,5R)-5-(4-amino-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4-fluoro-2-(hydroxymethyl)tetrahydrofuran-3-ol was protected with Bz-Cl and pyridine to afford N-(7-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)benzamide, then protected with TBS-Cl and imidazole in DMF and cooled to 0 °C before heating the reaction mixture to 40 °C. The resulting product is then reacted with N-(9-((2R,3R,4R,5R)-5-((bis(4-methoxyphenyl)(phenyl)methoxy)methyl)-4-((tert-butyldimethylsilyl)oxy)-3-fluorotetrahydrofuran-2-yl)-9H-purin-6-yl)-N-((E)-4-hydroxybut-2-en-1-yl)benzamide using DIAD and triphenylphosphine in THF, and the reaction is cooled to 0°C and the reaction mixture is allowed to warm to room temperature over time. The free OH group of the resulting product is then converted to a phosphonate ester using the reagent shown under basic conditions. The DMT protecting group of the resulting product is then removed using dichloroacetic acid, water, and aqueous sodium bicarbonate. The deprotected product is then reacted and forms an intramolecular connection to the phosphonate group as shown. The sulfur functional group is then added to the resulting product using sulfur and trimethylamine. The TBS protecting group is then removed using the reagent shown. The resulting product is then functionalized with a 2-nitrobenzyl group. Then the product therefrom is converted into phosphoramidite (diluted with acetonitrile) in dichloromethane. The phosphoramidite product is then formed intramolecularly connected by free OH groups, as shown in the figure. The connected product is then converted into sulfur-containing product. Benzenethiol, trimethylamine and dioxane are used to react the product therefrom. Then the product therefrom is deprotected to obtain the desired compound, which is used as starting material in reaction scheme 2 below.

[1236] Reaction Scheme 2

[1237]

[1238]

[1239]

[1240] Intermediate Example 18 Preparation of Intermediate 2b

[1241]

[1242]

[1243] To a solution of intermediate 1b (100 mg, 134 μmol, 1 eq) in DCM (6 mL) was added pyridine (318 mg, 4.02 mmol, 325 μL, 30 eq) and 2-[tert-butoxycarbonyl(methyl)amino]ethylphosgene (542 mg, 2.28 mmol, 17 eq). The mixture was stirred at 25 °C for 1 hour. The mixture was concentrated in vacuo. The residue was purified by preparative HPLC (column: C 18-4 150·30mm·5μm; mobile phase: [A: 0.1M water (TFA) B: acetonitrile]; B%: 15%-45%, 20 minutes) to give the desired product (50 mg, 47.7 μmol, 35.57% yield, TEA) as a white solid.

[1244] Intermediate Example 19

[1245] Preparation of intermediate 3b

[1246]

[1247] Intermediate 2b (100 mg, 95.4 μmol, 1.0 equiv., TEA) was dissolved in acetonitrile (0.5 mL) and H 2 To a solution of 4% paraformaldehyde (50 μL, 1% paraformaldehyde) in 4% paraformaldehyde (0.5 mL) was added TFA (544 mg, 4.77 mmol, 353 μL, 50 equiv). The mixture was stirred at 50° C. for 1 hour. The mixture was concentrated in vacuo to give the desired product (84 mg, 87.4 μmol, 91.63% yield, TFA) as a white solid.

[1248] Intermediate Example 20

[1249] Preparation of intermediate 5b

[1250]

[1251] To a solution of intermediate 3b (40 mg, 41.6 μmol, 1 eq., TFA) in DMF (0.2 mL) was added DIEA (16.1 mg, 125 μmol, 21.8 μL, 3.0 eq.) and intermediate 4b (40.3 mg, 62.5 μmol, 1.5 eq.). The mixture was stirred at 0 °C for 1 hour. The pH of the mixture was adjusted to about 6 with TFA and then concentrated in vacuo. The residue was purified by HPLC with H 2 The mixture was diluted with 1% ethyl acetate (5 mL) and the aqueous phase was extracted with ethyl acetate (5 mL) to remove by-products. The aqueous phase was freeze-dried to give the desired product (50 mg, 37.0 μmol, 88.81% yield) as a white solid.

[1252] Intermediate Example 21

[1253] Preparation of intermediate 6b

[1254]

[1255] Intermediate 5b (50 mg, 37.0 μmol, 1.0 equiv) was dissolved in acetonitrile (0.5 mL) and H 2 To the solution in 4% 4-nitropropene (0.5 mL) was added TFA (211 mg, 1.85 mmol, 137 μL, 50 equiv). The mixture was stirred at 50° C. for 2 h. The mixture was concentrated in vacuo. The residue was purified by preparative HPLC (column: C 18-4 150·30mm·5μm; mobile phase: [A: 0.1M water (TFA) B: acetonitrile]; B%: 10%-40%, 20 minutes) to give the desired product (20 mg, 14.6 μmol, 39.59% yield, TFA) as a white solid.

[1256] Intermediate Example 22

[1257] Preparation of intermediate 8b

[1258]

[1259] To a solution of intermediate 6b (9.00 mg, 6.59 μmol, 1.0 eq., TFA) in DMF (0.1 mL) was added DIEA (2.55 mg, 19.8 μmol, 3.44 μL, 3.0 eq.) and intermediate 7b (9.18 mg, 6.59 μmol, 1 eq.) at 0°C. The mixture was stirred at 20°C for 1 hour. The pH of the mixture was adjusted to about 6 with TFA at 0°C. The mixture was concentrated in vacuo to give the desired product (15 mg crude product) as a colorless oil.

[1260] Intermediate Example 23

[1261] Synthesis of 4-[[tert-Butyl(dimethyl)silyl]oxymethyl]aniline

[1262]

[1263] To a solution of (4-aminophenyl)methanol (5.00 g, 40.6 mmol, 1.0 eq) in DCM (50 mL) at 0°C, imidazole (4.15 g, 60.9 mmol, 1.5 eq) and TBSCl (7.34 g, 48.7 mmol, 5.97 mL, 1.2 eq) were added and then stirred at 25°C for 1 hour. The reaction mixture was stirred at 0°C by adding 100 mL of H 2 O, and then extracted with DCM (30 mL x 3). The combined organic layers were washed with 50 mL of brine and 2 SO 4 The residue was purified by flash silica gel chromatography ( 80g Purification by silica flash column, eluting with a gradient of 50 to 70% ethyl acetate / petroleum ether at 80 mL / min). 4-[[tert-butyl(dimethyl)silyl]oxymethyl]aniline (9.6 g, 40.44 mmol, 99.60% yield) was obtained as a colorless oil: 1 H NMR (400 MHz, CDCl 3 )δ7.13 (d, J=8.0 Hz, 2H), 6.67 (d, J=8.0 Hz, 2H), 4.64 (s, 2H), 0.94 (s, 9H), 0.10 (s, 6H); LC / MS [M+H] 238.2 (calculated); LC / MS [M+H] 238.1 (observed).

[1264] Intermediate Example 24

[1265] Synthesis of tert-butyl N-[(1S)-2-[4-[[tert-butyl(dimethyl)silyl]oxymethyl]anilino]-1-methyl-2-oxo-ethyl]carbamate

[1266]

[1267] To a solution of 4-[[tert-butyl(dimethyl)silyl]oxymethyl]aniline (9.34 g, 39.3 mmol, 1 eq) and (2S)-2-(tert-butoxycarbonylamino)propanoic acid (8.93 g, 47.2 mmol, 1.2 eq) in DCM (50 mL) and methanol (50 mL) was added EEDQ (29.2 g, 118 mmol, 3.0 eq) and then stirred at 25° C. for 1 hour. The reaction mixture was stirred at 0° C. by adding 100 mL of H 2 O quenched, then extracted with ethyl acetate (50 mL × 3). The combined organic layer was washed with 50 mL of brine and 2 SO 4 The residue was purified by flash silica gel chromatography ( 120g The mixture was purified by silica flash column with a gradient of 15 to 30% ethyl acetate / petroleum ether at 80 mL / min. Tert-butyl N-[(1S)-2-[4-[[tert-butyl(dimethyl)silyl]oxymethyl]anilino]-1-methyl-2-oxo-ethyl]carbamate was obtained as an orange oil (25 g, crude product).

[1268] Intermediate Example 25

[1269] Synthesis of tert-butyl N-[(2S)-2-[bis(tert-butoxycarbonyl)amino]propionyl-1]-N-[4-[[tert-butyl-1(dimethyl)silyl]oxymethyl]phenyl]carbamate

[1270]

[1271] To a solution of tert-butyl N-[(1S)-2-[4-[[tert-butyl(dimethyl)silyl]oxymethyl]anilino]-1-methyl-2-oxo-ethyl]carbamate (10.0 g, 24.5 mmol, 1.0 equiv) in DCM (100 mL) were added DMAP (2.99 g, 24.5 mmol, 1.0 equiv), DIEA (9.49 g, 73.4 mmol, 12.8 mL, 3.0 equiv) and Boc 2 O (16.0 g, 73.4 mmol, 16.9 mL, 3.0 equiv). The mixture was stirred at 40 °C for 24 h. The reaction mixture was heated at 0 °C by adding 100 mL of H 2 O quenched, and then extracted with DCM (50 mL x 2). The combined organic layers were washed with 100 mL of brine, 2 SO 4 The residue was purified by flash silica gel chromatography ( 120g Silica flash column, eluting with a gradient of 10 to 20% ethyl acetate / petroleum ether at 80 mL / min) was used to purify. Tert-butyl N-[(2S)-2-[bis(tert-butoxycarbonyl)amino]propanoyl-1]-N-[4-[[tert-butyl-1(dimethyl)silyl]oxymethyl]phenyl]carbamate (4.70 g, 7.72 mmol, 31.54% yield) was obtained as a colorless oil: 1 HNMR (CDCl 3 , 400 MHz) δ7.31 (d, J = 8.4 Hz, 2H), 7.11 (d, J = 8.4 Hz, 2H), 5.48-5.43 (m, 1H), 4.75 (s, 2H), 1.55-1.50 (m, 27H), 1.02 (d, J = 6.4 Hz, 3H), 0.94 (s, 9H), 0.08 (s, 6H); LC / MS [M+Na] 631.35 (calculated); LC / MS [M+Na] 631.3 (observed).

[1272] Intermediate Example 26

[1273] Synthesis of tert-butyl N-[(2S)-2-[bis(tert-butoxycarbonyl)amino]propionyl]-N-[4-(hydroxymethyl)phenyl]carbamate

[1274]

[1275] To a solution of tert-butyl N-[(2S)-2-[bis(tert-butoxycarbonyl)amino]propanoyl]-N-[4-[[tert-butyl(dimethyl)silyl]oxymethyl]phenyl]carbamate (4.68 g, 7.69 mmol, 1.0 equiv) in THF (50 mL) was added TBAF (1 M, 15.4 mL, 2.0 equiv) and then stirred at 25° C. for 1 hour. The reaction mixture was heated at 0° C. by adding H 2 O (100 mL) was quenched, and then extracted with ethyl acetate (50 mL × 3). The combined organic layer was washed with 50 mL of brine and 2 SO 4 The residue was purified by flash silica gel chromatography ( 40g Purification was performed by silica flash column with a gradient of 50 to 70% ethyl acetate / petroleum ether at 60 mL / min. tert-Butyl N-[(2S)-2-[bis(tert-butoxycarbonyl)amino]propanoyl]-N-[4-(hydroxymethyl)phenyl]carbamate (2.10 g, 4.25 mmol, 55.24% yield) was obtained as a colorless oil: 1H NMR (400 MHz, CDCl 3 )δ7.36(d,J=8.4Hz,2H),7.15(d,J=8.4Hz,2H),5.49-5.44(m,1H),4.70(d,J=2.8Hz,2H),1.53(s,21H),1.35(s,9H); LC / MS[M+H]517.2(calculated); LC / MS[M+H]517.2(observed).

[1276] Intermediate Example 27

[1277] [4-[[(2S)-2-[bis(tert-butoxycarbonyl)amino]propionyl]-tert-butoxycarbonylamino]phenyl]methyl(4-nitrophenyl)

[1278] Synthesis of Carbonate

[1279]

[1280] To a solution of tert-butyl N-[(2S)-2-[bis(tert-butoxycarbonyl)amino]propanoyl]-N-[4-(hydroxymethyl)phenyl]carbamate (1.00 g, 2.02 mmol, 1.0 equiv) in DCM (15 mL) was added DIEA (784 mg, 6.07 mmol, 1.06 mL, 3.0 equiv) and bis(4-nitrophenyl)carbonate (1.23 g, 4.04 mmol, 2.0 equiv), and then stirred at 25° C. for 1 hour. The reaction mixture was stirred at 0° C. by adding H 2 O (30 mL) and then extracted with DCM (10 mL x 3). The combined organic layers were washed with 20 mL of brine and purified by Na 2 SO 4 The residue was purified by flash silica gel chromatography ( 20g The mixture was purified by silica flash column with a gradient of 10 to 30% ethyl acetate / petroleum ether at 40 mL / min. [4-[[(2S)-2-[bis(tert-butoxycarbonyl)amino]propanoyl]-tert-butoxycarbonylamino]phenyl]methyl(4-nitrophenyl)carbonate (1.71 g, crude product) was obtained as a colorless oil: LC / MS [M+Na] 682.3 (calculated); LC / MS [M+Na] 682.2 (observed).

[1281] Intermediate Example 28

[1282] Synthesis of tert-butyl N-[(2S)-2-[bis(tert-butoxycarbonyl)amino]propionyl]-N-[4-(ethylcarbamoyloxymethyl)phenyl]carbamate

[1283]

[1284] To a solution of ethylamine (315 mg, 3.87 mmol, 457 μL, 1.5 eq., HCl) in DMF (15 mL) was added DIEA (1.67 g, 12.9 mmol, 2.24 mL, 5.0 eq.) and [4-[[(2S)-2-[bis(tert-butoxycarbonyl)amino]propanoyl]-tert-butoxycarbonyl-amino]phenyl]methyl (4-nitrophenyl)carbonate (1.70 g, 2.58 mmol, 1.0 eq.) at 0°C. The mixture was stirred at 25°C for 1 hour. The reaction mixture was heated at 0°C by adding H 2 O (20 mL) was quenched, and then extracted with ethyl acetate (20 mL×3). The combined organic layers were washed with 20 mL of brine and purified by Na 2 SO 4 The residue was purified by flash silica gel chromatography ( 20g The mixture was purified by silica flash column with a gradient of 30 to 50% ethyl acetate / petroleum ether at 40 mL / min. tert-Butyl N-[(2S)-2-[bis(tert-butoxycarbonyl)amino]propanoyl]-N-[4-(ethylcarbamoyloxymethyl)phenyl]carbamate (1.14 g, 2.02 mmol, 78.21% yield) was obtained as a white solid: LC / MS [M+Na] 588.3 (calculated); LC / MS [M+Na] 588.3 (observed).

[1285] Intermediate Example 29

[1286] Synthesis of tert-butyl N-[(2S)-2-[bis(tert-butoxycarbonyl)amino]propionyl]-N-[4-[[chloromethyl(ethyl)carbamoyl]oxymethyl]phenyl]carbamate

[1287]

[1288] To a solution of tert-butyl N-[(2S)-2-[bis(tert-butoxycarbonyl)amino]propanoyl]-N-[4-(ethylcarbamoyloxymethyl)phenyl]carbamate (1.00 g, 1.77 mmol, 1.0 equiv) in toluene (10 mL) was added paraformaldehyde (3.15 g, 2.65 mmol, 1.5 equiv) and 1-chloro-N,N,2-trimethylprop-1-en-1-amine (1.18 g, 8.84 mmol, 1.17 mL, 5.0 equiv). The mixture was stirred at 80 °C for 1 hour. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The compound tert-butyl N-[(2S)-2-[bis(tert-butoxycarbonyl)amino]propanoyl]-N-[4-[[chloromethyl(ethyl)carbamoyl]oxymethyl]phenyl]carbamate (2 g, crude product) was obtained as a colorless oil.

[1289] Intermediate Example 30

[1290] N-[(2S)-2-[bis(tert-butoxycarbonyl)amino]propanoyl]-N-[4-[[[2-[(1

[1291] S,2S,4R,6R,8S,9S,11S,12R,13S,19S)-12,19-difluoro-6-(3-fluorophenyl)-11-hydroxy-9,13-dimethyl-16-oxo-5,7-dioxapentacyclo[10.8.0.02,9.04,8.013,18]eicosane-14,17-dien-8-yl]-2-oxo-ethoxy]methyl

[1292] Synthesis of tert-butyl [ethyl-carbamoyl]oxymethyl]phenyl]carbamate

[1293]

[1294] To a solution of tert-butyl N-[(2S)-2-[bis(tert-butoxycarbonyl)amino]propanoyl]-N-[4-[[chloromethyl(ethyl)carbamoyl]oxymethyl]phenyl]carbamate (1.18 g, 1.93 mmol, 2.0 equiv) in DCM (15 mL) was added DIEA (371 mg, 2.87 mmol, 0.5 mL, 2.98 equiv) and (1S,2S,4R,6R,8S,9S, 11S,12R,13S,19S)-12,19-difluoro-6-(3-fluorophenyl)-11-hydroxy-8-(2-hydroxyacetyl)-9,13-dimethyl-5,7-dioxapentacyclo[10.8.0.02,9.04,8.013,18]eicosane-14,17-dien-16-one (500 mg, 964.28 μmol, 1.0 equiv) was added and then stirred at 25° C. for 12 hours. The reaction mixture was heated at 0° C. by adding H2 O (50 mL) and then extracted with DCM (20 mL x 3). The combined organic layers were washed with 20 mL of brine and purified by Na 2 SO 4 The residue was purified by flash silica gel chromatography ( 20g The mixture was purified by silica flash column with a gradient of 50 to 70% ethyl acetate / petroleum ether at 45 mL / min. N-[(2S)-2-[bis(tert-butoxycarbonyl)amino]propanoyl]-N-[4-[[[2-[(1S,2S,4R,6R,8S,9S,11S,12R,13S,19S)-12,19-difluoro-6-(3-fluorophenyl)-11-hydroxy-9,13-dimethyl-16-oxo-5,7-dioxolane[10.8.0.02, 9.04,8.013,18]eicosane-14,17-dien-8-yl]-2-oxo-ethoxy]methyl-ethyl-carbamoyl]oxymethyl]phenyl]carbamic acid tert-butyl ester (658 mg, 600.26 μmol, 62.25% yield): LC / MS [M+Na] 1118.5 (calculated); LC / MS [M+Na] 1118.3 (observed).

[1295] Intermediate Example 31

[1296] Synthesis of N-[[2-[(1S,2S,4R,6R,8S,9S,11S,12R,13S,19S)-12,19-difluoro-6-(3-fluorophenyl)-11-hydroxy-9,13-dimethyl-16-oxo-5,7-dioxapentacyclo[10.8.0.02,9.04,8.013,18]eicosane-14,17-dien-8-yl]-2-oxo-ethoxy]methyl]-N-ethyl-carbamic acid [4-[[(2S)-2-aminopropionyl]amino]phenyl]methyl ester

[1297]

[1298] To a solution of tert-butyl N-[(2S)-2-[bis(tert-butoxycarbonyl)amino]propanoyl]-N-[4-[[[2-[(1S,2S,4R,6R,8S,9S,11S,12R,13S,19S)-12,19-difluoro-6-(3-fluorophenyl)-11-hydroxy-9,13-dimethyl-16-oxo-5,7-dioxopentacyclo[10.8.0.02,9.04,8.013,18]eicosane-14,17-dien-8-yl]-2-oxo-ethoxy]methyl-ethyl-carbamoyl]oxymethyl]phenyl]carbamate (0.96 g, 875 μmol, 1.0 equiv) in toluene (15 mL) was added SiO 2 (10.0 g, 166 mmol, 190 equiv.). The mixture was stirred at 120°C for 1 hour. The reaction mixture was filtered and SiO 2 It was washed with methanol (20 mL x 2), and the filtrate was concentrated under reduced pressure to obtain a residue. N-[[2-[(1S,2S,4R,6R,8S,9S,11S,12R,13S,19S)-12,19-difluoro-6-(3-fluorophenyl)-11-hydroxy-9,13-dimethyl-16-oxo-5,7-dioxacyclo[10.8.0.02,9.04,8.013,18]eicos-14,17-dien-8-yl]-2-oxo-ethoxy]methyl]-N-ethyl-carbamic acid [4-[[(2S)-2-aminopropanoyl]amino]phenyl]methyl ester (500 mg, 628.27 μmol, 71.74% yield) was obtained as a colorless oil: LC / MS [M+H] 796.3 (calculated); LC / MS [M+H] 796.4 (observed).

[1299] Intermediate Example 32

[1300] N-[[2-[(1S,2S,4R,6R,8S,9S,11S,12R,13S,19S)-12,19-difluoro-6-(3-fluorophenyl)-11-hydroxy-9,13-dimethyl-16-oxo-5,7-dioxapentacyclo[10.8.0.02,9.04,8.013,18]eicosane-14,17-dien-8-yl]-2-oxo-ethoxy]methyl]-N-ethyl-carbamic acid [4-[[(2S)-2-[[(2S)-2-amino-3-methyl-butanoyl]amino]propanoyl]amino]

[1301] Synthesis of phenyl]methyl ester

[1302]

[1303] To N-[[2-[(1S,2S,4R,6R,8S,9S,11S,12R,13S,19S)-12,19-difluoro-6-(3-fluorophenyl)-11-hydroxy-9,13-dimethyl-16-oxo-5,7-dioxapentacyclo[10.8.0.02,9.04,8.013,18]eicosane-14,17-dien-8-yl]-2-oxo-ethoxy]methyl]-N-ethyl-carbamic acid [4-[[(2S)-2 To a solution of [2-aminopropionyl]amino]phenyl]methyl ester (200 mg, 251 μmol, 1.0 equiv) in THF (2 mL) was added DIEA (97.4 mg, 754 μmol, 131 μL, 3.0 equiv) and (2,5-dioxopyrrolidin-1-yl)(2S)-2-(9H-fluoren-9-ylmethoxycarbonylamino)-3-methyl-butanoate (164 mg, 377 μmol, 1.5 equiv), and then stirred at 25 ° C for 1 hour. Piperidine (86.2 mg, 1.01 mmol, 0.10 mL, 5.66 equiv) was added. The mixture was stirred at 25 ° C for another 1 hour. The mixture was purified by preparative HPLC (column: Phenomenex Luna 80×30 mm×3 μm; mobile phase: [water (TFA)-ACN]; B%: 30%-65%, 8 minutes). N-[[2-[(1S,2S,4R,6R,8S,9S,11S,12R,13S,19S)-12,1 9-Difluoro-6-(3-fluorophenyl)-11-hydroxy-9,13-dimethyl-16-oxo-5,7-dioxapentacyclo[10.8.0.02,9.04,8.013,18]eicosane-14,17-dien-8-yl]-2-oxo-ethoxy]methyl]-N-ethyl-carbamic acid [4-[[(2S)-2-[[(2S)-2-amino-3-methyl-butanoyl]amino]propanoyl]amino]phenyl]methyl ester (100 mg, 44.69 μmol, 24.97% yield, 40% purity): LC / MS [M+H] 895.4 (calculated); LC / MS [M+H] 895.5 (observed).

[1304] Intermediate Example 33

[1305] Synthesis of (((9H-fluoren-9-yl)methoxy)carbonyl)-L-alanyl-L-alanylglycine tert-butyl ester

[1306]

[1307] To a stirred solution of (2S)-2-[(2S)-2-{[(9H-fluoren-9-ylmethoxy)carbonyl]amino}propionamido]propanoic acid (1.0 g, 2.6 mmol, 1.0 equiv) in N,N-dimethylformamide (5.0 mL) at 0° C. under an inert nitrogen atmosphere were added tert-butyl 2-aminoacetate (343.0 mg, 2.6 mmol, 1.0 equiv), N,N-diisopropylethylamine (675.9 mg, 5.2 mmol, 2.0 equiv), benzotriazol-1-yl-hexafluorophosphate oxytripyrrolidinophosphonium (530.0 mg, 3.9 mmol, 1.5 equiv). The resulting solution was stirred at 25° C. for 4 hours and diluted with water (100 mL). The resulting mixture was extracted with dichloromethane (3×100 mL), and the organic layer was washed with saturated sodium chloride solution (3×100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by flash preparative HPLC under the following conditions (IntelFlash-1): column, C 18 Silica gel; mobile phase, water and acetonitrile (10.0% acetonitrile increased to 100.0% in 20 minutes); detector, UV 254nm. 1.0 g (77%) of (((9H-fluoren-9-yl)methoxy)carbonyl)-L-alanyl-L-alanylglycine tert-butyl ester was obtained as a white solid. MS m / z [M+H] + (ESI):496.23.

[1308] Intermediate Example 34

[1309] Synthesis of (((9H-fluoren-9-yl)methoxy)carbonyl)-L-alanyl-L-alanylglycine

[1310]

[1311] At 25 ° C, under an inert nitrogen atmosphere, trifluoroacetic acid (10.0 mL) was added to a solution of (((9H-fluorene-9-yl)methoxy)carbonyl)-L-alanyl-L-alanylglycine tert-butyl ester (1.0 g, 2.0 mmol, 1.0 equiv) in dichloromethane (20.0 mL). The resulting solution was stirred at 25 ° C for 5 hours and concentrated in vacuo. The crude product was purified by flash preparative HPLC under the following conditions (IntelFlash-1): column, C 18 Silica gel; mobile phase, water and acetonitrile (10.0% acetonitrile increased to 100.0% in 20 minutes); detector, UV 254nm. 800 mg (90%) of (((9H-fluoren-9-yl)methoxy)carbonyl)-L-alanyl-L-alanylglycine was obtained as a yellow oil. MS m / z [MH] - (ESI):438.17.

[1312] Intermediate Example 35

[1313] Synthesis of (5S,8S)-1-(9H-fluoren-9-yl)-5,8-dimethyl-3,6,9-trioxo-2-oxa-4,7,10-triazaundecane-11-yl acetate

[1314]

[1315] To a stirred solution of (((9H-fluoren-9-yl)methoxy)carbonyl)-L-alanyl-L-alanylglycine (800.0 mg, 1.8 mmol, 1.0 equiv) and copper acetate monohydrate (33.0 mg, 0.2 mmol, 0.1 equiv) in N,N-dimethylformamide (10.0 mL) was added acetic acid (164.0 mg, 2.7 mmol, 1.5 equiv) and lead tetraacetate (4.0 g, 9.0 mmol, 5.0 equiv) at room temperature under air atmosphere. The resulting mixture was stirred at 60 ° C for another 5 hours. The resulting solution was cooled to 25 ° C and diluted with water (100 mL). The resulting mixture was extracted with dichloromethane (3×100 mL), and the organic layer was washed with saturated sodium chloride solution (3×100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by flash preparative HPLC under the following conditions (IntelFlash-1): column, C 18 Silica gel; mobile phase, water and acetonitrile (10.0% acetonitrile increased to 100.0% in 20 minutes); detector, UV 254nm. 425 mg (51%) of (5S,8S)-1-(9H-fluoren-9-yl)-5,8-dimethyl-3,6,9-trioxo-2-oxa-4,7,10-triazaundecane-11-yl acetate was obtained as a white solid. MS m / z [M+Na] + (ESI):476.17.

[1316] Intermediate Example 36

[1317] Synthesis of (5S,8S)-1-(9H-fluoren-9-yl)-5,8-dimethyl-3,6,9-trioxo-2-oxa-4,7,10-triazaundecane-11-yl acetate

[1318]

[1319] To the reaction mixture was added (5S,8S)-1-(9H-fluoren-9-yl)-5,8-dimethyl-3,6,9-trioxo-2-oxa-4,7,10-triazaundec-11-yl acetate (425.0 mg, 0.94 mmol, 1.0 equiv) and (2S,6aS,6bR,7S,8aS,8bS,10R,11aR,12aS,12bS)-2,6b-difluoro-10-(3-fluorophenyl)-7-hydroxy-8b-( To a stirred solution of 2-hydroxyacetyl)-6a,8a-dimethyl-1,2,6a,6b,7,8,8a,8b,11a,12,12a,12b-dodecahydro-4H-naphtho[2',1':4,5]indeno[1,2-d][1,3]dioxol-4-one (243.0 mg, 0.5 mmol, 0.5 equiv) in dichloromethane (5.0 mL) was added p-toluenesulfonic acid (32.2 mg, 0.2 mmol, 0.20 equiv) in portions. The resulting mixture was stirred at 25 °C for another 12 hours. The crude product was purified by reverse phase flash evaporation with the following conditions (column, C 18 Silica gel; mobile phase, acetonitrile in water (10 mmol / L ammonium bicarbonate), gradient from 10% to 100% in 20 minutes; detector, UV 254 nm). 200 mg (47%) of ((S)-1-(((S)-1-(((2-((2S,6aS,6bR,7S,8aS,8bS,10R,11aR,12aS,12bS)-2,6b-difluoro-10-(3-fluorophenyl)-7-hydroxy-6a,8a-dimethyl-4-oxo-1,2,4,6a ,6b,7,8,8a,11a,12,12a,12b-dodecahydro-8bH-naphtho[2',1':4,5]indeno[1,2-d][1,3]dioxol-8b-yl)-2-oxoethoxy)methyl)amino)-1-oxopropan-2-yl)amino)-1-oxopropan-2-yl)carbamic acid (9H-fluoren-9-yl)methyl ester. MS m / z[M+H] + (ESI):912.36.

[1320] Intermediate Example 37

[1321] (5S,8S)-1-(9H-fluoren-9-yl)-5,8-dimethyl-3,6,9-trioxo-2-oxa-4,7,10-triazaundecane-11-ylacetic acid

[1322] Synthesis of esters

[1323]

[1324] ((S)-1-(((S)-1-(((2-((2S,6aS,6bR,7S,8aS,8bS,10R,11aR,12aS,12bS)-2,6b-difluoro-10-(3-fluorophenyl)-7-hydroxy-6a,8a-dimethyl-4-oxo-1,2,4,6a,6b,7,8,8a,11a,12,12a,12b-dodecahydro-8 To a stirred solution of (9H-fluoren-9-yl)methyl ester (200 mg, 0.2 mmol, 1.0 equiv.) of ((1H-naphtho[2',1':4,5]indeno[1,2-d][1,3]dioxol-8b-yl)-2-oxoethoxy)methyl)amino)-1-oxopropan-2-yl)amino)-1-oxopropan-2-yl)carbamate (200 mg, 0.2 mmol, 1.0 equiv.) in N,N-dimethylformamide (2.0 mL) was added morpholine (0.3 mL). The resulting mixture was stirred at 25 °C for another hour. The crude product was purified by reverse phase flash distillation with the following conditions (column, C 18 Silica gel; mobile phase, acetonitrile in water (0.1% trifluoroacetic acid), gradient 10% to 100% in 20 minutes; detector, UV 254 nm). 100 mg (66%) of (S)-2-amino-N-((S)-1-(((2-((2S,6aS,6bR,7S,8aS,8bS,10R,11aR,12aS,12bS)-2,6b-difluoro-10-(3-fluorophenyl)-7-hydroxy-6a,8a-dimethyl-4-oxo-1,2,4,6a,6b,7,8,8a,11a,12,12a,12b-dodecahydro-8bH-naphtho[2′,1′:4,5]indeno[1,2-d][1,3]dioxol-8b-yl)-2-oxoethoxy)methyl)amino)-1-oxopropan-2-yl)propanamide were obtained as a white solid. MS m / z [M+H] + (ESI):690.29.

[1325] Intermediate Example 38

[1326] Synthesis of (((9H-fluoren-9-yl)methoxy)carbonyl)-L-phenylalanylglycine tert-butyl ester

[1327]

[1328] To a solution of (2S)-2-{[(9H-fluoren-9-ylmethoxy)carbonyl]amino}-3-phenylpropanoic acid (5.0 g, 12.9 mmol, 1.0 equiv) in N,N-dimethylformamide (50.0 mL) at 0° C. under an inert nitrogen atmosphere were added tert-butyl 2-aminoacetate hydrochloride (2.6 g, 15.5 mmol, 1.2 equiv), benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (10.1 g, 19.4 mmol, 1.5 equiv), 1-hydroxybenzotriazole (2.6 g, 19.4 mmol, 1.5 equiv), N,N-diisopropylethylamine (4.9 g, 38.7 mmol, 3.0 equiv). The resulting solution was stirred at 25° C. for 4 hours and diluted with water (1000 mL). The resulting mixture was extracted with dichloromethane (3×1000 mL), and the organic layer was washed with saturated sodium chloride solution (3×1000 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by flash preparative HPLC under the following conditions (IntelFlash-1): column, C 18 Silica gel; mobile phase, water and acetonitrile (10.0% acetonitrile increased to 100.0% in 20 minutes); detector, UV 254nm. 5.2 g (80%) of (((9H-fluoren-9-yl)methoxy)carbonyl)-L-phenylalanylglycine tert-butyl ester was obtained as a white solid. MS m / z [M+H] + (ESI):501.23.

[1329] Intermediate Example 39

[1330] Synthesis of L-phenylalanine tert-butyl ester

[1331]

[1332] Under an inert nitrogen atmosphere, to a solution of (((9H-fluorene-9-yl)methoxy)carbonyl)-L-phenylalanylglycine tert-butyl ester (5.2 g, 10.4 mmol, 1.0 equiv) in morpholine / N,N-dimethylformamide (7.0 mL / 49.0 mL), the resulting solution was stirred at 25 ° C for 1 hour and diluted with water (1000 mL). The resulting mixture was extracted with dichloromethane (3×1000 mL), and the organic layer was washed with saturated sodium chloride solution (3×1000 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by flash preparative HPLC under the following conditions (IntelFlash-1): column, C 18Silica gel; mobile phase, water and acetonitrile (10.0% acetonitrile increased to 100.0% in 20 minutes); detector, UV 254nm. 2.7 g (93%) of L-phenylalanylglycine tert-butyl ester was obtained as a white solid. MS m / z [M+H] + (ESI):279.16.

[1333] Intermediate Example 40

[1334] Synthesis of (((9H-fluoren-9-yl)methoxy)carbonyl)-glycylglycyl-L-phenylalanylglycine tert-butyl ester

[1335]

[1336] To a solution of L-phenylalanylglycine tert-butyl ester (2.7 g, 9.7 mmol, 1.0 equiv) in N,N-dimethylformamide (30.0 mL) at 0° C. under an inert nitrogen atmosphere, (2-{[(9H-fluoren-9-ylmethoxy)carbonyl]amino}acetamido)acetic acid (3.4 g, 9.7 mmol, 1.0 equiv), benzotriazol-1-yl-hexafluorophosphate oxytripyrrolidinophosphonium (7.6 g, 14.6 mmol, 1.5 equiv), 1-hydroxybenzotriazole (1.9 g, 14.6 mmol, 1.5 equiv), N,N-diisopropylethylamine (3.8 g, 29.1 mmol, 3.0 equiv) were added. The resulting solution was stirred at 25° C. for 4 hours and diluted with water (500 mL). The resulting mixture was extracted with dichloromethane (3×500 mL), and the organic layer was washed with saturated sodium chloride solution (3×500 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by flash preparative HPLC under the following conditions (IntelFlash-1): column, C 18 Silica gel; mobile phase, water and acetonitrile (10.0% acetonitrile increased to 100.0% in 20 minutes); detector, UV 254nm. 3.8 g (64%) of (((9H-fluoren-9-yl)methoxy)carbonyl)glycylglycyl-L-phenylalanylglycine tert-butyl ester was obtained as a white solid. MS m / z [M+H] + (ESI):615.27.

[1337] Intermediate Example 40

[1338] Synthesis of (((9H-fluoren-9-yl)methoxy)carbonyl)glycylglycyl-L-phenylalanylglycylglycine tert-butyl ester

[1339]

[1340] To a solution of (((9H-fluoren-9-yl)methoxy)carbonyl)glycylglycyl-L-phenylalanylglycine tert-butyl ester (3.8 g, 6.2 mmol, 1.0 equiv) in dichloromethane (40.0 mL) was added trifluoroacetic acid (10.0 mL) under an inert nitrogen atmosphere at 0° C. The resulting solution was stirred at 25° C. for 5 hours and concentrated in vacuo. To this solution in N,N-dimethylformamide (30.0 mL) at 0°C was added tert-butyl 2-aminoacetate hydrochloride (1.0 g, 6.2 mmol, 1.0 equiv), benzotriazol-1-yl-hexafluorophosphate oxytripyrrolidinophosphonium (4.8 g, 9.3 mmol, 1.5 equiv), 1-hydroxybenzotriazole (1.3 g, 9.3 mmol, 1.5 equiv), N,N-diisopropylethylamine (2.4 g, 18.6 mmol, 3.0 equiv). The resulting solution was stirred at 25°C for 4 hours and diluted with water (300 mL). The resulting mixture was extracted with dichloromethane (3×500 mL), and the organic layer was washed with saturated sodium chloride solution (3×500 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by flash preparative HPLC under the following conditions (IntelFlash-1): column, C 18 Silica gel; mobile phase, water and acetonitrile (10.0% acetonitrile increased to 100.0% in 20 minutes); detector, UV 254nm. 2.6 g (64%) of (((9H-fluoren-9-yl)methoxy)carbonyl)glycylglycyl-L-phenylalanylglycylglycine tert-butyl ester was obtained as a white solid. MS m / z [M+H] + (ESI): 672.30; 1 H NMR (300 MHz, DMSO-d 6 )δ:1.39(s,9H),2.75-2.82(m,1H),3.03-3.09(m,1H),3.56-3.80(m,8H),4.19-4.31(m,3H),4.49-4.57(m,1H),7.15-7.24 (m,7H),7.25-7.44(m,2H),7.57-7.61(m,1H),7.69-7.72(m,2H),7.88-7.99(m,2H),8.01-8.15(m,3H),8.31-8.35(m,1H).

[1341] Intermediate Example 40

[1342] Synthesis of (((9H-fluoren-9-yl)methoxy)carbonyl)glycylglycyl-L-phenylalanylglycylglycine

[1343]

[1344] At 0 ° C, under an inert nitrogen atmosphere, trifluoroacetic acid (15.0 mL) was added to a solution of (((9H-fluorene-9-yl)methoxy)carbonyl)glycylglycyl-L-phenylalanylglycylglycine tert-butyl ester (2.6 g, 3.9 mmol, 1.0 equiv) in dichloromethane (30.0 mL). The resulting solution was stirred at 25 ° C for 5 hours and concentrated in vacuo. The crude product was purified by flash preparative HPLC under the following conditions (IntelFlash-1): column, C 18 Silica gel; mobile phase, water (containing 0.1% trifluoroacetic acid) and acetonitrile (10.0% acetonitrile rising to 100.0% in 20 minutes); detector, UV 254 nm. 2.1 g (88%) of (((9H-fluoren-9-yl)methoxy)carbonyl)glycylglycyl-L-phenylalanylglycylglycine was obtained as a white solid. MS m / z [M+H] + (ESI):616.20.

[1345] Intermediate Example 41

[1346] Synthesis of (S)-11-Benzyl-1-(9H-fluoren-9-yl)-3,6,9,12,15-pentaoxo-2-oxa-4,7,10,13,16-pentaazaheptadecan-17-yl acetate

[1347]

[1348] To a solution of (((9H-fluoren-9-yl)methoxy)carbonyl)glycylglycyl-L-phenylalanylglycylglycine (2.1 g, 3.4 mmol, 1.0 equiv) in N,N-dimethylformamide (30.0 mL) at 25°C under an inert nitrogen atmosphere, anhydrous copper acetate (254 mg, 1.4 mmol, 0.4 equiv), acetic acid (468 mg, 7.8 mmol, 2.3 equiv), lead tetraacetate (1.8 g, 4.1 mmol, 1.2 equiv) were added. The resulting mixture was stirred at 60°C for 5 hours. The mixture was cooled to 25°C, diluted with water (300 mL), extracted with dichloromethane (3×500 mL), and the organic layer was washed with saturated sodium chloride solution (3×500 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by flash preparative HPLC under the following conditions (IntelFlash-1): column, C 18Silica gel; mobile phase, water and acetonitrile (10.0% acetonitrile increased to 100.0% in 20 minutes); detector, UV 254nm. 870 mg (41%) of (S)-11-benzyl-1-(9H-fluoren-9-yl)-3,6,9,12,15-pentaoxo-2-oxa-4,7,10,13,16-pentaazaheptadecan-17-yl acetate was obtained as a white solid. MS m / z [M+H] + (ESI):630.20.

[1349] Intermediate Example 42

[1350] ((S)-10-Benzyl-1-((2S,6aS,6bR,7S,8aS,8bS,10R,11aR,12aS,12bS)-2,6b-difluoro-10-(3-fluorophenyl)-7-hydroxy-6a,8a-dimethyl-4-oxo-1,2,4,6a,6b,7,8,8a,11a,12,12a,12b-dodecahydro-8bH-naphtho[2',1':4,5]indeno[1,2-d][1,3]dioxol-8b-yl)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecane

[1351] Synthesis of Methyl (9H-fluoren-9-yl)carbamate

[1352]

[1353] To a solution of (S)-11-benzyl-1-(9H-fluoren-9-yl)-3,6,9,12,15-pentaoxo-2-oxa-4,7,10,13,16-pentaazaheptadecan-17-yl (870.0 mg, 1.38 mmol, 3.0 equiv) in dichloromethane (8.0 mL) was added (2S,6aS,6bR,7S,8aS,8bS,10R,11aR,12aS,12bS)-2,6b-difluoro-10- (3-Fluorophenyl)-7-hydroxy-8b-(2-hydroxyacetyl)-6a,8a-dimethyl-1,2,6a,6b,7,8,8a,8b,11a,12,12a,12b-dodecahydro-4H-naphtho[2',1':4,5]indeno[1,2-d][1,3]dioxol-4-one (238.8 mg, 0.46 mmol, 1.0 equiv) was added followed by p-toluenesulfonic acid (31.7 mg, 0.18 mmol, 0.4 equiv) at 25°C. The resulting mixture was stirred at 40°C for 12 hours. The mixture was cooled to 25°C, diluted with water (300 mL), extracted with dichloromethane (3×500 mL), and the organic layer was washed with saturated sodium chloride solution (3×500 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by flash preparative HPLC under the following conditions (IntelFlash-1): column, C 18 Silica gel; mobile phase, water and acetonitrile (10.0% acetonitrile increased to 100.0% in 20 minutes); detector, UV 254nm. 150.0 mg (30%) of ((S)-10-benzyl-1-((2S,6aS,6bR,7S,8aS,8bS,10R,11aR,12aS,12bS)-2,6b-difluoro-10-(3-fluorophenyl)-7-hydroxy-6a,8a-dimethyl-4-oxo-1,2,4,6a,6b,7 ,8,8a,11a,12,12a,12b-dodecahydro-8bH-naphtho[2',1':4,5]indeno[1,2-d][1,3]dioxol-8b-yl)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecane-16-yl)(9H-fluoren-9-yl)carbamic acid methyl ester. MS m / z[M+H] + (ESI):1088.42.

[1354] Intermediate Example 43

[1355] Synthesis of (S)-2-(2-(2-aminoacetamido)acetamido)-N-(2-(((2-((2S,6AS,6BR,7S,8AS,8BS,10R,11AR,12AS,12BS)-2,6B-difluoro-10-(3-fluorophenyl)-7-hydroxy-6A,8A-dimethyl-4-oxo-1,2,4,6A,6B,7,8,8A,11A,12,12A,12B-dodecahydro-8BH-naphtho[2',1':4,5]indeno[1,2-D][1,3]dioxol-8B-yl)-2-oxoethoxy)methyl)amino)-2-oxoethyl)-3-phenylpropanamide

[1356]

[1357] Under an inert nitrogen atmosphere, ((S)-10-benzyl-1-((2S,6aS,6bR,7S,8aS,8bS,10R,11aR,12aS,12bS)-2,6b-difluoro-10-(3-fluorophenyl)-7-hydroxy-6a,8a-dimethyl-4-oxo-1,2,4,6a,6b,7,8,8a,11a,12,12a,12b-dodecahydro-8bH-naphtho[2',1':4,5]indeno[ 1,2-d][1,3]dioxol-8b-yl)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecane-16-yl)(9H-fluoren-9-yl)carbamic acid methyl ester (150.0 mg, 0.14 mmol, 1.0 equiv) in morpholine / N,N-dimethylformamide (0.1 mL / 0.7 mL), the resulting solution was stirred at 25 ° C for 1 hour and diluted with water (10 mL). The resulting mixture was extracted with dichloromethane (3×30 mL), and the organic layer was washed with saturated sodium chloride solution (3×30 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by flash preparative HPLC under the following conditions (IntelFlash-1): column, C 18Silica gel; mobile phase, water and acetonitrile (10.0% acetonitrile rising to 100.0% in 20 minutes); detector, UV 254 nm. 100.6 mg (84%) of (S)-2-(2-(2-aminoacetamido)acetamido)-N-(2-(((2-((2S,6aS,6bR,7S,8aS,8bS,10R,11aR,12aS,12bS)-2,6b-difluoro-10-(3-fluorophenyl)-7-hydroxy-6a,8a-dimethyl-4-oxo-1,2,4,6a,6b,7,8,8a,11a,12,12a,12b-dodecahydro-8bH-naphtho[2′,1′:4,5]indeno[1,2-d][1,3]dioxol-8b-yl)-2-oxoethoxy)methyl)amino)-2-oxoethyl)-3-phenylpropanamide were obtained as a white solid. MS m / z [M+H] + (ESI):866.35.

[1358] Intermediate Example 44

[1359] Synthesis of (2S,6AS,6BR,7S,8AS,8BS,10R,11AR,12AS,12BS)-2,6B-difluoro-10-(3-fluorophenyl)-7-hydroxy-8B-(2-hydroxyacetyl)-6A,8A-dimethyl-1,2,6A,6B,7,8,8A,8B,11A,12,12A,12B-dodecahydro-4H-naphtho[2',1':4,5]indeno[1,2-D][1,3]dioxol-4-one

[1360]

[1361] Under an inert nitrogen atmosphere, magnesium sulfate (2.0 g, 16.9 mmol, 3.5 equiv) and 3-fluorobenzaldehyde (903.0 mg, 7.28 mmol, 1.5 equiv) were added to a solution of fluocinolone (2.0 g, 4.85 mmol, 1.0 equiv) in acetonitrile (20.0 mL), followed by trifluoromethanesulfonic acid (2.2 g, 14.6 mmol, 3.0 equiv) at 0 ° C. The resulting solution was stirred at 0 ° C for 3 hours and the pH was adjusted to 7 with saturated sodium bicarbonate solution (1.0 mL). The mixture was diluted with 300 mL of water and extracted with 3×500 mL of dichloromethane. The organic layers were combined, washed with 3×500 mL of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product (>1 g) was purified by achiral SFC under the following conditions: column, Torus 2-PIC column 4.6×100 mm, 5 μm; mobile phase: isopropanol (1% 2 mol / L NH 3 -methanol) and CO 2; Detector, UV 254nm. 800mg (32%) of (2S,6aS,6bR,7S,8aS,8bS,10R,11aR,12aS,12bS)-2,6b-difluoro-10-(3-fluorophenyl)-7-hydroxy-8b-(2-hydroxyacetyl)-6a,8a-dimethyl-1,2,6a,6b,7,8,8a,8b,11a,12,12a,12b-dodecahydro-4H-naphtho[2',1':4,5]indeno[1,2-d][1,3]dioxol-4-one was obtained as a white solid: MS m / z [M+H] + (ESI): 519.20; 1 H NMR (400 MHz, methanol-d 4 )δ:0.99(s,3H),1.28-1.65(m,4H),1.71-1.83(m,3H),2.24-2.39(m,3H),2.66-2.74(m,1H),4.31-4.36(m,2H),4.65(d,J=19 .6Hz,1H),5.08-5.09(m,1H),5.49-5.62(m,2H),6.32-6.35(m,2H),7.09-7.18(m,2H),7.27-7.33(m,2H),7.38-7.41(m,1H).

[1362] Intermediate Example 45

[1363] Synthesis of (9H-fluoren-9-yl)methyl (2-(((((2-((2S,6AS,6BR,7S,8AS,8BS,10R,11AR,12AS,12BS)-2,6B-difluoro-10-(3-fluorophenyl)-7-hydroxy-6A,8A-dimethyl-4-oxo-1,2,4,6A,6B,7,8,8A,11A,12,12A,12B-dodecahydro-8BH-naphtho[2',1':4,5]indeno[1,2-D][1,3]dioxol-8B-yl)-2-oxoethoxy)(hydroxy)phosphoryl)oxy)(hydroxy)phosphoryl)oxy)ethyl)carbamate

[1364]

[1365] Under an inert nitrogen atmosphere, trimethylamine (91.0 mg, 0.90 mmol, 1.2 equiv) and 1,1'-carbonyldiimidazole (183.0 mg, 1.13 mmol, 1.5 equiv) were added to a solution of (9H-fluoren-9-yl)methyl (2-(phosphonooxy)ethyl)carbamate (327.0 mg, 0.90 mmol, 1.5 equiv) in N,N-dimethylformamide (5.0 mL). The resulting mixture was stirred at 25° C. for 30 minutes. To this mixture was added 2-((2S,6aS,6bR,7S,8aS,8bS,10R,11aR,12aS,12bS)-2,6b-difluoro-10-(3-fluorophenyl)-7-hydroxy-6a,8a-dimethyl-4-oxo-1,2,4,6a,6b,7,8,8a,11a,12,12a,12b-dodecahydro-8bH-naphtho[2',1':4,5]indeno[1,2-d][1,3]dioxol-8b-yl)-2-oxoethyl dihydrogen phosphate (450.0 mg, 0.75 mmol, 1.0 equiv) and zinc chloride (816.0 mg, 6.0 mmol, 8.0 equiv). The resulting mixture was stirred at 25°C overnight, diluted with methanol (10.0 mL) and filtered. The filtrate was concentrated under reduced pressure. The crude product was purified by flash evaporation under the following conditions: column, C 18 Silica gel; mobile phase, water (containing 5 mmol / L ammonium hydroxide) and acetonitrile (10.0% acetonitrile increased to 100.0% in 20 minutes); detector, UV 254nm. 310 mg of (9H-fluoren-9-yl)methyl (2-(((((2-((2S,6aS,6bR,7S,8aS,8bS,10R,11aR,12aS,12bS)-2,6b-difluoro-10-(3-fluorophenyl)-7-hydroxy-6a,8a-dimethyl-4-oxo-1,2,4,6a,6b,7,8,8a,11a,12,12a,12b-dodecahydro-8bH-naphtho[2′,1′:4,5]indeno[1,2-d][1,3]dioxol-8b-yl)-2-oxoethoxy)(hydroxy)phosphoryl)oxy)(hydroxy)phosphoryl)oxy)ethyl)carbamate were obtained as a white solid. MS m / z [MH] - (ESI):942.20.

[1366] Intermediate Example 46

[1367] Synthesis of (2-(((((2-((2S,6AS,6BR,7S,8AS,8BS,10R,11AR,12AS,12BS)-2,6B-difluoro-10-(3-fluorophenyl)-7-hydroxy-6A,8A-dimethyl-4-oxo-1,2,4,6A,6B,7,8,8A,11A,12,12A,12B-dodecahydro-8BH-naphtho[2',1':4,5]indeno[1,2-D][1,3]dioxol-8B-yl)-2-oxoethoxy)(hydroxy)phosphoryl)oxy)(hydroxy)phosphoryl)oxy)ethyl)carbamate

[1368]

[1369] Under an inert nitrogen atmosphere, (2-(((((2-((2S,6aS,6bR,7S,8aS,8bS,10R,11aR,12aS,12bS)-2,6b-difluoro-10-(3-fluorophenyl)-7-hydroxy-6a,8a-dimethyl-4-oxo-1,2,4,6a,6b,7,8,8a,11a,12,12a,12b-dodecahydro-8bH-naphtho[2' To a solution of (1,1':4,5]indeno[1,2-d][1,3]dioxol-8b-yl)-2-oxoethoxy)(hydroxy)phosphoryl)oxy)(hydroxy)phosphoryl)oxy)ethyl)carbamic acid (9H-fluoren-9-yl)methyl ester (310.0 mg, 0.33 mmol, 1.0 equiv) in dichloromethane (5.0 mL) was added piperidine (196.0 mg, 2.31 mmol, 7.0 equiv). The resulting solution was stirred at 25 ° C for 3 hours and concentrated under reduced pressure. The crude product was purified by flash evaporation under the following conditions: column, C 18 Silica gel; mobile phase, water (containing 5 mmol / L ammonium hydroxide) and acetonitrile (5.0% acetonitrile increased to 100.0% in 20 minutes); detector, UV 254nm. The eluent was concentrated under reduced pressure. Under an inert nitrogen atmosphere, Dowex 50w × 8 (200.0 mg) was added to a solution of the desired product (ammonium salt) in water (5.0 mL). The resulting solution was stirred at 25 ° C for another hour and filtered. The filtrate was concentrated under reduced pressure. 115 mg (21% over two steps) of the desired product was obtained as a white solid. MS m / z [MH] - (ESI):720.20.

[1370] Intermediate Example 47

[1371] Synthesis of (E)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl acrylate

[1372]

[1373] To a stirred solution of copper(I) chloride (0.09 g, 0.93 mmol, 0.03 eq.) and 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (0.54 g, 0.93 mmol, 0.03 eq.) in tetrahydrofuran (50 mL) was added sodium tert-butoxide (0.18 g, 1.87 mmol, 0.06 eq.) at 0°C. The reaction solution was stirred at 0°C for 1 hour. To the above mixture was added 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1,3,2-dioxaborolane (15.85 g, 62.43 mmol, 2.0 eq.) in portions at room temperature. The resulting mixture was stirred at room temperature for another 1 hour. At room temperature, benzyl prop-2-ynoate (5.0 g, 31.22 mmol, 1.0 equiv) and methanol (1.5 g, 46.82 mmol, 1.5 equiv) were added to the above mixture in batches. The resulting mixture was stirred for another 14 hours at 25 ° C. The aqueous layer was extracted with dichloromethane (3 × 200 mL). The residue was passed through silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether / ethyl acetate = 1 / 0, 3 / 1). 5.0 g (55%) of the desired product was obtained as a colorless liquid.

[1374] Intermediate Example 48

[1375] Synthesis of tert-butyl 4-(2-bromo-5-nitrophenoxy)butyrate

[1376]

[1377] Potassium carbonate (9.5 g, 68.8 mmol, 3 equiv) was added in batches to a stirred solution of 2-bromo-5-nitrophenol (5.0 g, 22.9 mmol, 1.0 equiv) and tert-butyl 4-bromobutyrate (7.7 g, 34.40 mmol, 1.5 equiv) in N,N-dimethylformamide (100.0 mL) at 25 ° C under a nitrogen atmosphere. The resulting mixture was stirred for 3 hours at 25 ° C under a nitrogen atmosphere and diluted with water (500 mL). The resulting mixture was extracted with dichloromethane (3×500 mL), and the organic layer was washed with a saturated sodium chloride solution (3×500 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by flash preparative HPLC under the following conditions (IntelFlash-1): column, C 18Silica gel; mobile phase, water and acetonitrile (10.0% acetonitrile rising to 100.0% in 20 minutes); detector, UV 254 nm. 5 g (60%) of tert-butyl 4-(2-bromo-5-nitrophenoxy)butyrate were obtained as a yellow solid.

[1378] Intermediate Example 49

[1379] Synthesis of tert-butyl (E)-4-(2-(3-(benzyloxy)-3-oxoprop-1-en-1-yl)-5-nitrophenoxy)butyrate

[1380]

[1381] To tert-butyl 4-(2-bromo-5-nitrophenoxy)butyrate (5.0 g, 13.88 mmol, 1.0 equiv), benzyl (2E)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)prop-2-enoate (4.8 g, 16.65 mmol, 1.2 equiv) and tripotassium phosphate (5.89 g, 27.76 mmol, 2.0 2-Dicyclohexylphosphino-2',6'-dimethoxy-1,1'-biphenyl (0.28 g, 0.69 mmol, 0.05 equiv) and tris(dibenzylideneacetone)dipalladium (0.64 g, 0.69 mmol, 0.05 equiv) were added in portions to a stirred solution of N,N-dimethylformamide (50.0 mL), and the reaction solution was stirred at 90 ° C for another 12 hours under a nitrogen atmosphere. The resulting mixture was cooled to 25 ° C and diluted with water (500 mL). The resulting mixture was extracted with dichloromethane (3×500 mL), and the organic layer was washed with saturated sodium chloride solution (3×500 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by flash preparative HPLC under the following conditions (IntelFlash-1): column, C 18 Silica gel; mobile phase, water and acetonitrile (10.0% acetonitrile rising to 100.0% in 20 minutes); detector, UV 254 nm. 3 g (49%) of (E)-tert-butyl 4-(2-(3-(benzyloxy)-3-oxoprop-1-en-1-yl)-5-nitrophenoxy)butanoate were obtained as a yellow oil.

[1382] Intermediate Example 50

[1383] Synthesis of 3-(4-amino-2-(4-(tert-butoxy)-4-oxobutoxy)phenyl)propionic acid

[1384]

[1385] To a solution of tert-butyl (E)-4-(2-(3-(benzyloxy)-3-oxoprop-1-en-1-yl)-5-nitrophenoxy)butanoate (3.0 g, 6.80 mmol, 1.0 equiv) in methanol (30.0 mL) was added palladium on carbon (1.0 g). The flask was evacuated and flushed with hydrogen five times. The resulting solution was stirred at 25 °C for 3 hours, the solid was filtered out and washed with methanol (3×10 mL). The resulting mixture was concentrated under reduced pressure, and the crude product was used directly in the next step without further purification. 2.5 g (81%) of 3-(4-amino-2-(4-(tert-butoxy)-4-oxobutoxy)phenyl)propanoic acid was obtained as a white solid.

[1386] Intermediate Example 51

[1387] 3-(2-(4-(tert-Butyloxy)-4-oxobutoxy)-4-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)phenyl)

[1388] Synthesis of Propionic Acid

[1389]

[1390] To a solution of 3-(4-amino-2-(4-(tert-butoxy)-4-oxobutoxy)phenyl)propanoic acid (2.5 g, 7.73 mmol, 1.0 equiv) and methyl 2,5-dioxopyrrole-1-carboxylate (1.80 g, 11.59 mmol, 1.5 equiv) in dichloromethane (25.0 mL) was added trimethylamine (1.56 g, 15.46 mmol, 2.0 equiv) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 50 ° C for 4 hours. The resulting mixture was cooled to 25 ° C and concentrated under reduced pressure. 1.5 g (48%) of 3-(2-(4-(tert-butoxy)-4-oxobutoxy)-4-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)phenyl)propanoic acid was obtained as a yellow oil. MS m / z [MH] - (ESI):402.16.

[1391] Intermediate Example 52

[1392] 4-(5-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-(3-oxo-3-(2,3,5,6-tetrafluorophenoxy)propyl)phenoxy)

[1393] Synthesis of tert-butyl butyrate

[1394]

[1395] A solution of 3-(2-(4-(tert-butoxy)-4-oxobutoxy)-4-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)phenyl)propanoic acid (1.5 g, 3.71 mmol, 1 eq) in N,N-dimethylformamide (15 mL) was treated with 2,3,5,6-tetrafluorophenol (1.85 g, 11.15 mmol, 3 eq) at room temperature under nitrogen atmosphere, followed by the addition of 1-ethyl-3(3-dimethylpropylamine)carbodiimide (2.14 g, 11.15 mmol, 3 eq) at 25 °C. The resulting mixture was stirred at 25 °C for 2 hours under nitrogen atmosphere. The residue was purified by reverse phase flash chromatography under the following conditions: column, C 18 Silica gel; mobile phase, acetonitrile in water (0.1% formic acid), gradient from 10% to 100% in 10 minutes; detector, UV 254 nm. The resulting mixture was concentrated under reduced pressure. 800 mg (39%) of tert-butyl 4-(5-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-(3-oxo-3-(2,3,5,6-tetrafluorophenoxy)propyl)phenoxy)butanoate was obtained as a colorless oil.

[1396] Intermediate Example 53

[1397] 4-(5-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-(3-oxo-3-(2,3,5,6-tetrafluorophenoxy)propyl)phenoxy)

[1398] Synthesis of butyric acid

[1399]

[1400] At 25 ° C, under an inert nitrogen atmosphere, trifluoroacetic acid (2.0 mL) was added to a solution of tert-butyl 4-(5-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-(3-oxo-3-(2,3,5,6-tetrafluorophenoxy)propyl)phenoxy)butanoate (800 mg, 1.45 mmol, 1 equivalent) in dichloromethane (6.0 mL). The resulting mixture was stirred at 25 ° C for 2 hours. The resulting mixture was concentrated under reduced pressure. 700 mg (97%) of 4-(5-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-(3-oxo-3-(2,3,5,6-tetrafluorophenoxy)propyl)phenoxy)butanoic acid was obtained as a yellow oil.

[1401] Intermediate Example 54

[1402] Synthesis of (2,3,5,6-tetrafluorophenyl) 3-[2-chlorocarbonyl-4-(2,5-dioxypyrrol-1-yl)phenyl] propionate

[1403]

[1404] 1-Chloro-N, N, 2-trimethyl-prop-1-ene-1-amine (1.10 g, 8.23 ​​mmol, 1.09 mL, 2.0 equiv) was added to a solution of 5-(2,5-dioxopyrrole-1-yl)-2-[3-oxo-3-(2,3,5,6-tetrafluorophenoxy)propyl]benzoic acid (1.80 g, 4.12 mmol, 1.0 equiv) in DCM (36 mL). The mixture was stirred at 25 ° C for 1 hour. The reaction mixture was concentrated under reduced pressure to remove the solvent. It was used directly in the next step without further purification. Compound (2,3,5,6-tetrafluorophenyl) 3-[2-chlorocarbonyl-4-(2,5-dioxopyrrole-1-yl) phenyl] propionate (1.88 g, 4.13 mmol, 100% yield) was obtained as a yellow oil.

[1405] Intermediate Example 55

[1406] Synthesis of 2-[[2-[[2-[[2-[[2-[[2-[[2-[[2-[[2-[[2-[[2-[[2-[[2-[[5-(2,5-dioxopyrrol-1-yl)-2-[3-oxo-3-(2,3,5,6-tetrafluorophenoxy)propyl]benzoyl]-methyl-amino]acetyl]-methyl-amino]acetyl]-methyl-amino]acetyl]-methyl-amino]acetyl]-methyl-amino]acetyl]-methyl-amino]acetyl]-methyl-amino]acetyl]-methyl-amino]acetyl]-methyl-amino]acetyl]-methyl-amino]acetyl]-methyl-amino]acetyl]

[1407]

[1408] To a solution of 2-[methyl-[2-[methyl-[2-[methyl-[2-[methyl-[2-[methyl-[2-[methyl-[2-[methyl-[2-[methyl-[2-[methyl-[2-[methyl-[2-[methyl-[2-(methylamino)acetyl]amino]acetyl]amino]acetyl]amino]-acetyl]amino]acetyl]amino]acetyl]amino]acetyl]amino]acetyl]amino]acetyl]amino]acetyl]amino]acetyl]amino]acetyl]amino]acetyl]acetic acid (3.01 g, 4.13 mmol, 1.0 equiv) PSAR in DCM (36 mL) at 0 °C was added DIEA (1.60 g, 12.4 mmol, 2.16 mL, 3.0 equiv). After 10 minutes, to a solution of (2,3,5,6-tetrafluorophenyl) 3-[2-chlorocarbonyl-4-(2,5-dioxopyrrol-1-yl) phenyl] propanoate (1.88 g, 4.13 mmol, 1.0 equiv) in DCM (18 mL) was added and then stirred at 0°C for 1 hour. The pH of the resulting mixture was adjusted to about 6 with TFA at 0°C, diluted by adding MeCN (2 mL), and concentrated under reduced pressure to remove DCM. The residue was purified by preparative HPLC (column: Phenomenex Luna C 18 (250×70 mm, 15 μm; mobile phase: [water (TFA)-ACN]; B%: 25%-55%, 20 minutes) purification. 2-[[2-[[2-[[2-[[2-[[2-[[2-[[2-[[2-[[2-[[2-[[5-(2,5-dioxopyrrol-1-yl)-2-[3-oxo-3-(2,3,5,6-tetrafluorophenoxy)propyl]benzoyl]-methyl-amino]acetyl]-methyl-amino]acetyl]-methyl-amino]acetyl]-methyl-amino]acetyl]-methyl-amino]acetyl]-methyl-amino]acetyl]-methyl-amino]acetyl]-methyl-amino]acetyl]-methyl-amino]acetyl]-methyl-amino]acetyl]-methyl-amino]acetyl]-methyl-amino]acetyl]-methyl-amino]acetyl]-methyl-amino]acetyl]-methyl-amino]acetyl]acetic acid was obtained as a white solid (360 mg, 314 μmol, 7.60% yield): LC / MS [M+H] 1148.4 (calculated); LC / MS [M+H] 1148.5 (observed).

[1409] Intermediate Example 56

[1410] Synthesis of (2S,3S,4S,5R,6S)-2-(Methoxycarbonyl)-6-(2-nitro-4-((((4-nitrophenoxy)carbonyl)oxy)methyl)phenoxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate

[1411]

[1412] To a stirred solution of (2S,3S,4S,5R,6S)-3,4,5-tris(acetoxy)-6-[4-(hydroxymethyl)-2-nitrophenoxy]oxane-2-carboxylic acid methyl ester (5.0 g, 10.3 mmol, 1.0 equiv) and diisopropylethylamine (4.0 g, 31.0 mmol, 3.0 equiv) in N,N-dimethylformamide (50.0 mL) was added bis(4-nitrophenyl) carbonate (4.7 g, 15.5 mmol, 1.5 equiv) at room temperature under air atmosphere. The resulting solution was stirred at 25 °C for 16 hours and diluted with water (500 mL). The resulting mixture was extracted with dichloromethane (3×500 mL), and the organic layer was washed with saturated sodium chloride solution (3×500 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by flash preparative HPLC under the following conditions (IntelFlash-1): column, C 18 Silica gel; mobile phase, water and acetonitrile (10.0% acetonitrile increased to 100.0% in 20 minutes); detector, UV 254nm. 5.5 g (82%) of (2S,3S,4S,5R,6S)-2-(methoxycarbonyl)-6-(2-nitro-4-((((4-nitrophenoxy)carbonyl)oxy)methyl)phenoxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate was obtained as a white solid. MS m / z [M+NH 4 ] + (ESI):668.12.

[1413] Intermediate Example 57

[1414] (2S,3R,4S,5S,6S)-2-(4-(((ethylcarbamoyl)oxy)methyl)-2-nitrophenoxy)-6-(methoxycarbonyl)tetrahydro

[1415] Synthesis of 2H-pyran-3,4,5-triyl triacetate

[1416]

[1417] To a stirred solution of (2S,3S,4S,5R,6S)-2-(methoxycarbonyl)-6-(2-nitro-4-((((4-nitrophenoxy)carbonyl)oxy)methyl)phenoxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate (5.5 g, 8.5 mmol, 1.0 equiv) and ethylamine (1.4 g, 16.9 mmol, 2.0 equiv) in N,N-dimethylformamide (50.0 mL) at 25° C. under air atmosphere was added diisopropylethylamine (3.3 g, 25.4 mmol, 3.0 equiv). The resulting solution was stirred at 25° C. for 2 hours, diluted with water (500 mL). The resulting mixture was extracted with dichloromethane (3×500 mL), and the organic layer was washed with saturated sodium chloride solution (3×500 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by flash preparative HPLC under the following conditions (IntelFlash-1): column, C 18 Silica gel; mobile phase, water and acetonitrile (10.0% acetonitrile increased to 100.0% in 20 minutes); detector, UV 254nm. 3.5 g (75%) of (2S,3R,4S,5S,6S)-2-(4-(((ethylcarbamoyl)oxy)methyl)-2-nitrophenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate was obtained as a white solid. MS m / z [M+NH 4 ] + (ESI):574.15.

[1418] Intermediate Example 58

[1419] Synthesis of (2S,3R,4S,5S,6S)-2-(4-((((chloromethyl)(ethyl)carbamoyl)oxy)methyl)-2-nitrophenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate

[1420]

[1421] To a stirred solution of (2S,3R,4S,5S,6S)-2-(4-(((ethylcarbamoyl)oxy)methyl)-2-nitrophenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (3.5 g, 6.3 mmol, 1.0 equiv) and paraformaldehyde (1.7 g, 18.9 mmol, 3.0 equiv) in dichloromethane (35.0 mL) was added chlorotrimethylsilane (2.1 g, 18.9 mmol, 3.0 equiv) at 25° C. under air atmosphere. The resulting mixture was stirred at 25° C. for another 2 hours. The resulting mixture was concentrated under reduced pressure to give (2S,3R,4S,5S,6S)-2-(4-((((chloromethyl)(ethyl)carbamoyl)oxy)methyl)-2-nitrophenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (3.0 g, 79%) as a white solid.

[1422] Intermediate Example 59

[1423] (2S,3R,4S,5S,6S)-2-(4-(((((2-((2S,6AS,6BR,7S,8AS,8BS,10R,11AR,12AS,12BS)-2,6B-difluoro-10-(3-fluorophenyl)-7-hydroxy-6A,8A-dimethyl-4-oxo-1,2,4,6A,6B,7,8,8A,11A,12,12A,12B-dodecahydro-8BH-naphtho[2',1':4,5]indeno[1,2-D][1,3]dioxol-8B-yl)-2-oxoethoxy)methyl)(ethyl)carbamoyl)oxy)methyl)-2-nitrophenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate

[1424] become

[1425]

[1426] To a stirred solution of (2S,3R,4S,5S,6S)-2-(4-((((chloromethyl)(ethyl)carbamoyl)oxy)methyl)-2-nitrophenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (3.0 g, 5.0 mmol, 2 eq) and diisopropylethylamine (51.28 mg, 0.397 mmol, 3 eq) in N,N-dimethylformamide (20.0 mL) was added (2S,6aS, 6bR,7S,8aS,8bS,10R,11aR,12aS,12bS)-2,6b-difluoro-10-(3-fluorophenyl)-7-hydroxy-8b-(2-hydroxyacetyl)-6a,8a-dimethyl-1,2,6a,6b,7,8,8a,8b,11a,12,12a,12b-dodecahydro-4H-naphtho[2',1':4,5]indeno[1,2-d][1,3]dioxol-4-one (1.3 g, 2.5 mmol, 1.0 equiv). The resulting solution was stirred at 25 °C for 2 hours and diluted with water (300 mL). The resulting mixture was extracted with dichloromethane (3×300 mL), and the organic layer was washed with saturated sodium chloride solution (3×300 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by flash preparative HPLC under the following conditions (IntelFlash-1): column, C 18 Silica gel; mobile phase, water and acetonitrile (10.0% acetonitrile increased to 100.0% in 20 minutes); detector, UV 254 nm. 1.1 g (41%) of (2S,3R,4S,5S,6S)-2-(4-(((((2-((2S,6aS,6bR,7S,8aS,8bS,10R,11aR,12aS,12bS)-2,6b-difluoro-10-(3-fluorophenyl)-7-hydroxy-6a,8a-dimethyl-4-oxo-1,2,4,6a,6 b,7,8,8a,11a,12,12a,12b-dodecahydro-8bH-naphtho[2',1':4,5]indeno[1,2-d][1,3]dioxol-8b-yl)-2-oxoethoxy)methyl)(ethyl)carbamoyl)oxy)methyl)-2-nitrophenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate. MSm / z[M+H] + (ESI):1087.35.

[1427] Intermediate Example 60

[1428] Synthesis of (2S,3R,4S,5S,6S)-2-(2-amino-4-(((((2-((2S,6AS,6BR,7S,8AS,8BS,10R,11AR,12AS,12BS)-2,6B-difluoro-10-(3-fluorophenyl)-7-hydroxy-6A,8A-dimethyl-4-oxo-1,2,4,6A,6B,7,8,8A,11A,12,12A,12B-dodecahydro-8BH-naphtho[2',1':4,5]indeno[1,2-D][1,3]dioxol-8B-yl)-2-oxoethoxy)methyl)(ethyl)carbamoyl)oxy)methyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate

[1429]

[1430] At 25 °C, (2S,3R,4S,5S,6S)-2-(4-(((((2-((2S,6aS,6bR,7S,8aS,8bS,10R,11aR,12aS,12bS)-2,6b-difluoro-10-(3-fluorophenyl)-7-hydroxy-6a,8a-dimethyl-4-oxo-1,2,4,6a,6b,7,8,8a,11a,12,12a,12b-dodecahydro-8bH-naphtho[2',1':4,5]indeno[1,2- To a stirred solution of [1,3]dioxol-8b-yl)-2-oxoethoxy)methyl) (ethyl)carbamoyl)oxy)methyl)-2-nitrophenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (1.1 g, 1.0 mmol, 1.0 equiv) and ammonium chloride (536.8 mg, 10.1 mmol, 10.0 equiv) in methanol (11.0 mL) was added iron powder (1.1 g, 20.0 mmol, 20.0 equiv). The resulting mixture was stirred at 70 ° C for 5 hours. The solid was filtered out and washed with methanol (3×10 mL). The resulting mixture was concentrated under reduced pressure. The crude product was purified by flash preparative HPLC under the following conditions (IntelFlash-1): column, C 18Silica gel; mobile phase, water and acetonitrile (10.0% acetonitrile increased to 100.0% in 20 minutes); detector, UV 254nm. 810mg (76%) of (2S,3R,4S,5S,6S)-2-(2-amino-4-(((((2-((2S,6aS,6bR,7S,8aS,8bS,10R,11aR,12aS,12bS)-2,6b-difluoro-10-(3-fluorophenyl)-7-hydroxy-6a,8a-dimethyl-4-oxo-1,2, 4,6a,6b,7,8,8a,11a,12,12a,12b-Dodecahydro-8bH-naphtho[2',1':4,5]indeno[1,2-d][1,3]dioxol-8b-yl)-2-oxoethoxy)methyl)(ethyl)carbamoyl)oxy)methyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate. MS m / z[M+H] + (ESI):1057.37.

[1431] Intermediate Example 61

[1432] (2S,3R,4S,5S,6S)-2-(2-(3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propionamido)-4-(((((2-((2S,6AS,6BR,7S,8AS,8BS,10R,11AR,12AS,12BS)-2,6B-difluoro-10-(3-fluorophenyl)-7-hydroxy-6A,8A-dimethyl-4-oxo-1,2,4,6A,6B,7,8,8A,11A,12,12A,12B-dodecahydro-8BH-naphtho[2',1':4,5]indeno[1,2-D][1,3]dioxol-8B-yl)-2-oxoethoxy)methyl)(ethyl)carbamoyl)oxy)methyl)

[1433] Synthesis of (phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate

[1434]

[1435]

[1436] At 25 °C, (2S,3R,4S,5S,6S)-2-(2-amino-4-(((((2-((2S,6aS,6bR,7S,8aS,8bS,10R,11aR,12aS,12bS)-2,6b-difluoro-10-(3-fluorophenyl)-7-hydroxy-6a,8a-dimethyl-4-oxo-1,2,4,6a,6b,7,8,8a,11a,12,12a,12b-dodecahydro-8bH-naphtho[2',1':4,5]indeno[1,2-d][1,3]dioxole-8 To a stirred solution of (1,2-(2-((9H-fluoren-9-ylmethoxy)carbonyl]amino}propionic acid (357.8 mg, 1.2 mmol, 1.5 equiv) and ethyl 2-ethoxy-2H-quinoline-1-carboxylate (378.9 mg, 1.5 mmol, 2.0 equiv) in dichloromethane (10.0 mL) was added 3-{[(9H-fluoren-9-ylmethoxy)carbonyl]amino}propionic acid (357.8 mg, 1.2 mmol, 1.5 equiv). The resulting solution was stirred at 25 °C for 3 hours and diluted with water (200 mL). The resulting mixture was extracted with dichloromethane (3×300 mL), and the organic layer was washed with saturated sodium chloride solution (3×300 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by flash preparative HPLC under the following conditions (IntelFlash-1): column, C 18 Silica gel; mobile phase, water (10 mmol / L ammonium bicarbonate) and acetonitrile (10.0% acetonitrile increased to 100.0% in 20 minutes); detector, UV 254 nm. 600.0 mg (58%) of (2S,3R,4S,5S,6S)-2-(2-(3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propionamido)-4-(((((2-((2S,6aS,6bR,7S,8aS,8bS,10R,11aR,12aS,12bS)-2,6b-difluoro-10-(3-fluorophenyl)-7-hydroxy-6a,8 a-dimethyl-4-oxo-1,2,4,6a,6b,7,8,8a,11a,12,12a,12b-dodecahydro-8bH-naphtho[2',1':4,5]indeno[1,2-d][1,3]dioxol-8b-yl)-2-oxoethoxy)methyl)(ethyl)carbamoyl)oxy)methyl)phenoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate. MS m / z[M+H] + (ESI):1050.48.

[1437] Intermediate Example 62

[1438] Synthesis of (2S,3S,4S,5R,6S)-6-(2-(3-aminopropylamino)-4-(((((2-((2S,6AS,6BR,7S,8AS,8BS,10R,11AR,12AS,12BS)-2,6B-difluoro-10-(3-fluorophenyl)-7-hydroxy-6A,8A-dimethyl-4-oxo-1,2,4,6A,6B,7,8,8A,11A,12,12A,12B-dodecahydro-8BH-naphtho[2',1':4,5]indeno[1,2-D][1,3]dioxol-8B-yl)-2-oxoethoxy)methyl)(ethyl)carbamoyl)oxy)methyl)phenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid

[1439]

[1440] At 0°C under an inert nitrogen atmosphere, (2S,3R,4S,5S,6S)-2-(2-(3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propionamido)-4-(((((2-((2S,6aS,6bR,7S,8aS,8bS,10R,11aR,12aS,12bS)-2,6b-difluoro-10-(3-fluorophenyl)-7-hydroxy-6a,8a-dimethyl-4-oxo-1,2,4,6a,6b,7,8,8a,11a,12,12a To a stirred solution of (600.0 mg, 0.4 mmol, 1.0 equiv) in methanol (10.0 mL) and water (5.0 mL) was added lithium hydroxide (31.9 mg, 1.2 mmol, 3.0 equiv). The resulting solution was stirred at 25 °C for 12 hours and the pH was adjusted to 7 with hydrochloric acid (1 mol / L). The crude product was purified by flash preparative HPLC under the following conditions (IntelFlash-1): column, C 18Silica gel; mobile phase, water and acetonitrile (10.0% acetonitrile increased to 100.0% in 20 minutes); detector, UV 254nm. 180 mg (41%) of (2S,3S,4S,5R,6S)-6-(2-(3-aminopropylamino)-4-(((((2-((2S,6aS,6bR,7S,8aS,8bS,10R,11aR,12aS,12bS)-2,6b-difluoro-10-(3-fluorophenyl)-7-hydroxy-6a,8a-dimethyl-4- Oxo-1,2,4,6a,6b,7,8,8a,11a,12,12a,12b-dodecahydro-8bH-naphtho[2',1':4,5]indeno[1,2-d][1,3]dioxol-8b-yl)-2-oxoethoxy)methyl)(ethyl)carbamoyl)oxy)methyl)phenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid. MS m / z[M+H] + (ESI):988.36.

[1441] Intermediate Example 63

[1442] Synthesis of Methyl (E)-3-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)acrylate

[1443]

[1444] To a solution of copper(I) chloride (1.06 g, 10.7 mmol, 0.03 equiv) and 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (6.19 g, 10.7 mmol, 0.03 equiv) in tetrahydrofuran (450.0 mL) at 0°C was added sodium tert-butoxide (2.06 g, 21.4 mmol, 0.06 equiv). The reaction solution was stirred at 0°C for 1 hour, followed by the addition of a solution of bis(pinacolato)diboron (90.6 g, 357 mmol, 1.0 equiv) in tetrahydrofuran (150.0 mL). The reaction solution was stirred at 20°C under a nitrogen atmosphere for one hour. Methyl prop-2-ynoate (30.0 g, 357.0 mmol, 1.0 equiv) and methanol (22.9 g, 714 mmol, 2.0 equiv) were added to the above reaction solution. The reaction solution was stirred for another 12 hours at 20 ° C. The resulting mixture was poured into ice water (w / w=1 / 1) (300 mL) and stirred for 5 minutes. The aqueous phase was extracted with ethyl acetate (200 mL×3). The combined organic phase was washed with brine (50 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether / ethyl acetate=1 / 0, 3 / 1) to obtain (E)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)methyl acrylate (70 g, 92%) as a colorless oil: 1 H NMR (400 MHz, CDCl 3 )δ6.83-6.75(m,1H),6.68-6.59(m,1H),3.77(s,3H),1.29(s,12H).

[1445] Intermediate Example 64

[1446] Synthesis of (Benzyloxycarbonyl)glycylglycine tert-butyl ester

[1447]

[1448] To a solution of 2-(benzyloxycarbonylamino)acetic acid (5.50 g, 26.3 mmol, 1.0 equiv) in N,N-dimethylformamide (100.0 mL) was added triethylamine (7.98 g, 78.9 mmol, 3.0 equiv), 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (12.0 g, 31.5 mmol, 1.2 equiv) and tert-butyl 2-aminoacetate (3.45 g, 26.3 mmol, 1.0 equiv), and then stirred at 25° C. for 2 hours. The reaction mixture was quenched at 0° C. by adding water (150 mL), and then extracted with ethyl acetate (100 mL×3). The combined organic layers were washed with water (80 mL×3), washed with brine (100 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, ethyl acetate / petroleum ether=1 / 1) to give ((benzyloxy)carbonyl)glycylglycine tert-butyl ester (7.7 g, 91%) obtained as a yellow oil: 1 H NMR (CDCl 3 ,400MHz) δ7.40-7.31(m,5H),5.14(s,2H),3.98-3.89(m,4H),1.47(s,9H).

[1449] Intermediate Example 65

[1450] The synthesis of

[1451]

[1452] To a solution of ((benzyloxy)carbonyl)glycylglycine tert-butyl ester (8.8 g, 27.3 mmol, 1.0 equiv) in methanol (150 mL) was added palladium / carbon (10%, 2.5 g) under a nitrogen atmosphere. The suspension was degassed and purged with hydrogen 3 times. The mixture was stirred at 25 ° C under hydrogen (50 Psi) for 2 hours. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. Glycylglycine tert-butyl ester (4.2 g, 81%) was obtained as a white solid. MS m / z[M+H] + (ESI): 189.12; 1 HNMR (CDCl 3 ,400MHz) δ7.67(s,1H),3.96(d,J=5.6Hz,2H),3.39(s,2H),1.75(s,2H),1.46(s,9H).

[1453] Intermediate Example 66

[1454] Synthesis of tert-butyl 2-bromo-5-nitrobenzoate

[1455]

[1456] At 0 ° C, dicyclohexylcarbodiimide (8.95 g, 43.4 mmol, 1.1 equiv) and 4-dimethylaminopyridine (2.41 g, 19.7 mmol, 0.5 equiv) were added to a mixture of 2-bromo-5-nitrobenzoic acid (9.7 g, 39.4 mmol, 1.0 equiv) in dichloromethane (100 mL). The mixture was stirred at 0 ° C for 10 minutes, then tert-butyl alcohol (4.4 g, 59.1 mmol, 1.5 equiv) was added and stirred at 20 ° C for 12 hours. The mixture was poured into ice water (w / w=1 / 1) (50 mL) and stirred for 10 minutes. The aqueous phase was extracted with dichloromethane (500 mL×3). The combined organic phase was washed with brine (100 mL×3), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether / ethyl acetate=1 / 0, 5 / 1) to give tert-butyl 2-bromo-5-nitrobenzoate (9.2 g, 77%) as a white solid: 1 H NMR (400 MHz, CDCl 3 )δ8.52(d,J=2.8Hz,1H),8.13(dd,J=2.8,8.8Hz,1H),7.83(d,J=8.8Hz,1H),1.65(s,9H).

[1457] Intermediate Example 67

[1458] Synthesis of tert-butyl (E)-2-(3-methoxy-3-oxoprop-1-en-1-yl)-5-nitrobenzoate

[1459]

[1460] A mixture of (E)-methyl 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)acrylate (11.2 g, 53.0 mmol, 2.5 equiv), tert-butyl 2-bromo-5-nitrobenzoate (6.40 g, 21.2 mmol, 1.0 equiv), tripotassium phosphate (6.74 g, 31.8 mmol, 1.5 equiv), 2-dicyclohexylphosphino-2',6'-dimethoxy-1,1'-biphenyl (870 mg, 2.12 mmol, 0.1 equiv), tris(dibenzylideneacetone)dipalladium (970 mg, 1.06 mmol, 0.05 equiv) in dioxane (120.0 mL) and water (25.0 mL) was degassed and purged with nitrogen three times, and then the mixture was stirred at 90° C. under a nitrogen atmosphere for 12 hours. The mixture was poured into ice water (w / w=1 / 1) (30 mL) and stirred for 10 minutes. The aqueous phase was extracted with ethyl acetate (500 mL×3). The combined organic phase was washed with brine (100 mL×3), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether / ethyl acetate=1 / 0, 3 / 1) to obtain (E)-2-(3-methoxy-3-oxoprop-1-ene-1-yl)-5-nitrobenzoic acid tert-butyl ester (6 g, 92%) as a yellow solid: 1 H NMR (400 MHz, CDCl 3 )δ8.74(d,J=2.4Hz,1H),8.41(d,J=15.6Hz,1H),8.34(dd,J=2.4,8.4Hz,1H ), 7.72 (d, J = 8.4Hz, 1H), 6.38 (d, J = 15.6Hz, 1H), 3.85 (s, 3H), 1.65 (s, 9H).

[1461] Intermediate Example 68

[1462] Synthesis of tert-butyl 5-amino-2-(3-methoxy-3-oxopropyl)benzoate

[1463]

[1464] A mixture of (E)-tert-butyl 2-(3-methoxy-3-oxoprop-1-en-1-yl)-5-nitrobenzoate (4.0 g, 13.0 mmol, 1.0 equiv), palladium / carbon (10%, 400.0 mg) in methanol (50.0 mL) was degassed and purged with hydrogen three times, and then the mixture was stirred at 25° C. under a hydrogen atmosphere for 3 hours. The mixture was filtered and concentrated in vacuo to give tert-butyl 5-amino-2-(3-methoxy-3-oxopropyl)benzoate (3.50 g, 96%) as a yellow oil: [M+H] + (ESI): 280.30; 1 H NMR (400 MHz, CDCl 3 )δ7.14(d,J=2.4Hz,1H),7.04(d,J=8.0Hz,1H),6.73(dd,J=2.4,8.0Hz,1H),3.67(s,3H),3.12(t,J=8.0Hz,2H),2.61(t,J=8.0Hz,2H),1.59(s,9H).

[1465] Intermediate Example 69

[1466] Synthesis of 5-amino-2-(3-methoxy-3-oxopropyl)benzoic acid

[1467]

[1468] To a solution of tert-butyl 5-amino-2-(3-methoxy-3-oxopropyl)benzoate (3.5 g, 12.5 mmol, 1.0 equiv) in ethyl acetate (10.0 mL) was added hydrogen chloride (gas) / ethyl acetate (4 mol / L, 50 mL, 16.0 equiv) and then stirred at 25 °C for 12 hours. The mixture was concentrated in vacuo to give 5-amino-2-(3-methoxy-3-oxopropyl)benzoic acid (3.20 g, 98%) as a white solid: [M+H] + (ESI): 224.23; 1 H NMR (400 MHz, DMSO-d 6 )δ7.69(d,J=2.0Hz,1H),7.39-7.31(m,2H),3.57(s,3H),3.15(t,J=7.6Hz,2H),2.59(t,J=7.6Hz,2H).

[1469] Intermediate Example 70

[1470] Synthesis of methyl 3-(4-amino-2-((2-((2-(tert-butoxy)-2-oxoethyl)amino)-2-oxoethyl)carbamoyl)phenyl)propanoate

[1471]

[1472] At 0°C, 4-methylmorpholine (3.74 g, 37.0 mmol, 3.0 equiv.), 1-hydroxybenzotriazole (833 mg, 6.16 mmol, 0.5 equiv.), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (4.72 g, 24.7 mmol, 2.0 equiv.) and tert-butyl 2-[(2-aminoacetyl)amino]acetate (2.78 g, 14.8 mmol, 1.2 equiv.) were added to a solution of 5-amino-2-(3-methoxy-3-oxopropyl)benzoic acid (3.20 g, 12.3 mmol, 1.0 equiv.) in N,N-dimethylformamide (40 mL), and then stirred at 20°C for 2 hours. The mixture was poured into ice water (w / w=1 / 1) (40 mL) and stirred for 10 minutes. The aqueous phase was extracted with ethyl acetate (500 mL×3). The combined organic phases were washed with brine (200 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether / ethyl acetate = 1 / 0, 0 / 1) to give methyl 3-(4-amino-2-((2-((2-(tert-butoxy)-2-oxoethyl)amino)-2-oxoethyl)carbamoyl)phenyl)propanoate (3.8 g, 78%) as a yellow oil: [M+H] + (ESI): 394.35; 1 H NMR (400 MHz, CDCl 3 )δ7.04(d,J=8.0Hz,2H),6.79-6.66(m,3H),4.15(d,J=5.6Hz,2H),3.97(d,J=5. 2Hz, 2H), 3.61 (s, 3H), 2.99 (t, J = 7.2Hz, 2H), 2.68 (t, J = 7.2Hz, 2H), 1.47 (s, 9H).

[1473] Intermediate Example 71

[1474] Synthesis of (5-amino-2-(3-methoxy-3-oxypropyl)benzoyl)glycylglycine

[1475]

[1476] To a solution of methyl 3-(4-amino-2-((2-((2-(tert-butoxy)-2-oxoethyl)amino)-2-oxoethyl)carbamoyl)phenyl)propanoate (0.60 g, 1.53 mmol, 1.0 equiv) in ethyl acetate (5.0 mL) was added hydrogen chloride (gas) / ethyl acetate (4 mol / L, 10 mL, 26.2 equiv) and then stirred at 25 °C for 12 hours. The mixture was concentrated in vacuo to give (5-amino-2-(3-methoxy-3-oxopropyl)benzoyl)glycylglycine (550 mg, 96% yield) as a white solid: [M+H] + (ESI): 338.14; 1 H NMR (400 MHz, DMSO-d 6 )δ8.57(t,J=6.0Hz,1H),8.25(t,J=6.4Hz,1H),7.28(d,J=8.0Hz,1H),7.20-7.10(m,2H),3.89(d ,J=6.0Hz,2H),3.81(d,J=6.0Hz,2H),3.57(s,3H),2.90(t,J=8.0Hz,2H),2.60(t,J=8.0Hz,2H).

[1477] Intermediate Example 72

[1478] 1-(5-amino-2-(3-methoxy-3-oxypropyl)phenyl)-1,4,7-trioxo-11,14,17,20,23,26,29,32,35,38,41,44-

[1479] Synthesis of tert-butyl dodeca-2,5,8-triazaheptadecane-47-oate

[1480]

[1481] (5-amino-2-(3-methoxy-3-oxopropyl)benzoyl)glycylglycine (320.0 mg, 0.86 mmol, 1.0 equiv), tert-butyl 3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2-aminoethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propanoate (576.0 mg, 0 A mixture of 4-(2-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (328.0 mg, 1.71 mmol, 2.0 equivalents), 1-hydroxybenzotriazole (57.8 mg, 0.43 mmol, 0.5 equivalents) and 4-methylmorpholine (259.0 mg, 2.57 mmol, 3.0 equivalents) in N,N-dimethylformamide (5.0 mL) was stirred at 25 ° C for 2 hours. The reaction mixture was quenched at 0 ° C by adding water (10 mL), and then extracted with dichloromethane / isopropanol (v / v=3:1, 100 mL×5). The combined organic layer was concentrated under reduced pressure to give a residue. Column, C 18 Silica gel; mobile phase, acetonitrile in water (0.1% trifluoroacetic acid / formic acid), gradient from 10% to 70% in 20 minutes; detector, UV 254 nm. 950 mg (90%) of 1-(5-amino-2-(3-methoxy-3-oxopropyl)phenyl)-1,4,7-trioxo-11,14,17,20,23,26,29,32,35,38,41,44-dodeca-2,5,8-triazatetraheptadecane-47-oic acid tert-butyl ester was obtained as a yellow oil. [M+H] + (ESI):994.10.

[1482] Intermediate Example 73

[1483] Synthesis of (S)-3-(4-amino-2-((47-(tert-butoxycarbonyl)-52,52-dimethyl-2,5,45,50-tetraoxo-9,12,15,18,21,24,27,30,33,36,39,42,51-trideca-3,6,46-triazatripentyloxy)carbamoyl)phenyl)propionic acid

[1484]

[1485]

[1486] To a solution of tert-butyl 1-(5-amino-2-(3-methoxy-3-oxopropyl)phenyl)-1,4,7-trioxo-11,14,17,20,23,26,29,32,35,38,41,44-dodeca-2,5,8-triazatetraheptadecane-47-ate (850 mg, 956 μmol, 1.0 equiv) in dioxane (5.0 mL) was added hydrogen chloride (gas) / ethyl acetate (15 mL, 4 mol / L). The mixture was stirred at 25° C. for 2 hours. The reaction mixture was concentrated under reduced pressure. 1-(5-amino-2-(3-methoxy-3-oxopropyl)phenyl)-1,4,7-trioxo-11,14,17,20,23,26,29,32,35,38,41,44-dodeca-2,5,8-triazatetraheptadecane-47-oic acid (800 mg, crude product) was obtained as a yellow solid. [M+H] + (ESI):936.20.

[1487] 1-(5-amino-2-(3-methoxy-3-oxopropyl)phenyl)-1,4,7-trioxo-11,14,17,20,23,26,29,32,35,38,41,44-dodeca-2,5,8-triazatetraheptadecane-47-oic acid (800 mg, 0.903 mmol, 1.0 equivalent), (2S)-2-aminopentanedioic acid di-tert-butyl ester (588 mg, 1.99 mmol) 1, 2.2 eq., HCl), 1-ethyl-3 (3-dimethylpropylamine) carbodiimide (346 mg, 1.81 mmol, 2.0 eq.), 1-hydroxybenzotriazole (61.0 mg, 0.451 mmol, 0.5 eq.) and 4-methylmorpholine (274 mg, 2.71 mmol, 3.0 eq.) in N, N-dimethylformamide (10.0 mL) were stirred at 25 ° C for 2 hours. The reaction mixture was quenched at 0 ° C by adding water (20 mL) and then extracted with dichloromethane / isopropanol (v:v=3:1, 50 mL×5). The combined organic layers were concentrated under reduced pressure to give a residue. Column, C 18 Silica gel; mobile phase, acetonitrile in water (0.1% trifluoroacetic acid / formic acid), gradient from 10% to 70% in 20 minutes; detector, UV 254 nm. 900 mg (89%) of (1-(2-(5-amino-2-(3-methoxy-3-oxopropyl)benzamido)acetamido)-2-oxo-6,9,12,15,18,21,24,27,30,33,36,39-dodeca-3-azatetradodecane-42-yl)-L-glutamic acid di-tert-butyl ester was obtained as a yellow oil. [M+H] + (ESI):1179.05.

[1488] To a solution of (1-(2-(5-amino-2-(3-methoxy-3-oxopropyl)benzamido)acetamido)-2-oxo-6,9,12,15,18,21,24,27,30,33,36,39-dodeca-3-azatetradodecane-42-yl)-L-glutamic acid di-tert-butyl ester (800 mg, 0.68 mmol, 1.0 equiv) in methanol (8.0 mL) and water (2.5 mL), sodium hydroxide (136 mg, 3.39 mmol, 5.0 equiv) was added, and then stirred at 0° C. for 2 hours. The mixture was concentrated in vacuo to remove methanol, and then diluted with water (20 mL), the pH of the aqueous phase was adjusted to about 7 with hydrochloric acid (2 mol / L), and extracted with dichloromethane / isopropanol (v / v=3:1, 30 mL×5). The combined organic layers were dried over sodium sulfate, filtered and concentrated under reduced pressure to give a residue to afford (S)-3-(4-amino-2-((47-(tert-butoxycarbonyl)-52,52-dimethyl-2,5,45,50-tetraoxo-9,12,15,18,21,24,27,30,33,36,39,42,51-trideca-3,6,46-triazatripentyloxy)carbamoyl)phenyl)propanoic acid (700 mg, crude) as a red oil. [M+H] + (ESI):1165.10.

[1489] Intermediate Example 74

[1490] (1-(2-(5-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-(3-oxo-3-(2,3,5,6-tetrafluorophenoxy)propyl)benzamido)acetamido)-2-oxo-6,9,12,15,18,21,24,27,30,33,36,39-dodeca-3-azatetradecane

[1491] Synthesis of di-tert-butyl-4-(2-acyl)-L-glutamic acid

[1492]

[1493]

[1494] To a solution of (S)-3-(4-amino-2-((47-(tert-butoxycarbonyl)-52,52-dimethyl-2,5,45,50-tetraoxo-9,12,15,18,21,24,27,30,33,36,39,42,51-trideca-3,6,46-triazatripentyloxy)carbamoyl)phenyl)propanoic acid (700 mg, 0.60 mmol, 1.0 equiv) and methyl 2,5-dioxopyrrole-1-carboxylate (139.8 mg, 0.92 mmol, 1.5 equiv) in dichloromethane (7.0 mL) was added trimethylamine (182.2 mg, 1.80 mmol, 3.0 equiv) and then stirred at 50 °C for 4 hours. The pH of the mixture was adjusted to about 6 with trifluoroacetic acid, and diluted by adding water (10 mL), and extracted with dichloromethane:isopropanol (v / v=3:1, 30 mL×5). The combined organic layers were concentrated under reduced pressure to give a residue. Column, C 18 Silica gel; mobile phase, acetonitrile in water (0.1% trifluoroacetic acid / formic acid), gradient from 10% to 70% in 20 minutes; detector, UV 254 nm. 306 mg (41%) of (S)-3-(2-((47-(tert-butoxycarbonyl)-52,52-dimethyl-2,5,45,50-tetraoxo-9,12,15,18,21,24,27,30,33,36,39,42,51-trideca-3,6,46-triazatripentyloxy)carbamoyl)-4-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)phenyl)propanoic acid was obtained as a colorless oil. [M+H] + (ESI):1245.32.

[1495] 2,3,5,6-tetrafluorophenol (108 mg, 0.65 mmol, 3.0 equivalents), (S)-3-(2-((47-(tert-butoxycarbonyl)-52,52-dimethyl-2,5,45,50-tetraoxo-9,12,15,18,21,24,27,30,33,36,39,42,51-trideca-3,6,46-triazatripent A mixture of 4-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)phenyl)propanoic acid (270 mg, 216 μmol, 1.0 equiv) and 1-ethyl-3(3-dimethylpropylamine)carbodiimide (166 mg, 867 μmol, 4.0 equiv) in N,N-dimethylformamide (3.0 mL) was stirred at 25 °C for 2 hours. The reaction mixture was concentrated under reduced pressure. The crude product was purified by flash preparative HPLC under the following conditions (IntelFlash-1): column, C 18Silica gel; mobile phase, water and acetonitrile (10.0% acetonitrile increased to 100.0% in 20 minutes); detector, UV 254nm. 90 mg (30%) of (1-(2-(5-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-(3-oxo-3-(2,3,5,6-tetrafluorophenoxy)propyl)benzamido)acetamido)-2-oxo-6,9,12,15,18,21,24,27,30,33,36,39-dodeca-3-azatetradodecane-42-yl)-L-glutamic acid di-tert-butyl ester was obtained as a colorless oil. [M+H] + (ESI): 1394.45; 1 HNMR (DMSO-d 6 ,400MHz)δ7.94-7.84(m,1H),7.50(d,J=8.4Hz,1H),7.43-7.34(m,2H),7.18(s,2 H),4.13(dd,J=5.2,9.1Hz,1H),3.89(s,2H),3.70(s,2H),3.60-3.58(m,3H),3.5 1-3.47(m,44H),3.39(t,J=6.0Hz,2H),3.22-3.17(m,2H),3.14(s,3H),2.42-2.3 2(m,2H),2.27-2.19(m,2H),1.95-1.83(m,1H),1.77-1.65(m,1H),1.38(s,18H).

[1496] Intermediate Example 75

[1497] (1-(2-(5-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-(3-oxo-3-(2,3,5,6-tetrafluorophenoxy)propyl)benzamido)acetamido)-2-oxo-6,9,12,15,18,21,24,27,30,33,36,39-dodeca-3-azatetradecane

[1498] Synthesis of 4-(2-acyl)-L-glutamic acid

[1499]

[1500] To a solution of (1-(2-(5-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-(3-oxo-3-(2,3,5,6-tetrafluorophenoxy)propyl)benzamido)acetamido)-2-oxo-6,9,12,15,18,21,24,27,30,33,36,39-dodeca-3-azatetradodecane-42-yl)-L-glutamic acid di-tert-butyl ester (90 mg, 0.07 mmol, 1.0 equiv) in dichloromethane (1.0 mL) was added trifluoroacetic acid (0.25 mL) under an inert nitrogen atmosphere at 0° C. The resulting solution was stirred at 0° C. for 4 hours and concentrated under vacuum and reduced pressure. 90 mg (crude product) of (1-(2-(5-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-(3-oxo-3-(2,3,5,6-tetrafluorophenoxy)propyl)benzamido)acetamido)-2-oxo-6,9,12,15,18,21,24,27,30,33,36,39-dodeca-3-azatetradodecane-42-yl)-L-glutamic acid were obtained as a yellow oil. [M+H] + (ESI):1279.95.

[1501] Synthesis Example 1

[1502] Preparation of Compound I-15

[1503]

[1504] To a solution of intermediate 7 (55.0 mg, 26.2 μmol, 1.0 equiv.) in DCM (1 mL) was added TFA (1.54 g, 13.5 mmol, 1.00 mL, 513 equiv.). The mixture was stirred at 25 °C for 2 hours. The reaction mixture was concentrated in vacuo to give a residue, and the residue was purified by preparative HPLC (column: Phenomenex Luna 80 × 30 mm × 3 μm; mobile phase: [water (TFA)-acetonitrile]; B%: 15%-45%, 8 minutes). The eluent was removed under freeze drying. The desired product (8.50 mg, 4.04 μmol, 15.3% yield, 99.4% purity, TFA) was obtained as a white solid.

[1505] 1 H NMR (D 2O,400MHz)δ9.05(s,1H),8.31(s,1H),7.91-7.76(m,2H),7.63(d,J=8.6Hz,1H),7.46-7.16(m,7H),7.09(s,1H),6.93(s,2H),5.14(s,2 H),4.43-4.34(m,1H),4.32-4.24(m,1H),4.17-4.02(m,2H),3.97-3.79(m,5H),3.72(t,J=6.0Hz,2H),3.68-3.55(m,46H),3.53(t,J=6 .0Hz,2H),3.48-3.30(m,8H),3.16-3.11(m,2H),3.08-2.97(m,4H),2.66-2.61(m,2H),2.58-2.48(m,2H),2.42(t,J=7.2Hz,2H),2.21- 2.07(m,1H),1.98-1.85(m,2H),1.85-1.75(m,1H),1.73-1.56(m,5H),1.54-1.34(m,2H),0.89(t,J=7.2Hz,3H),0.79(t,J=6.0Hz,9H).

[1506] HPLC: 99.45% (220nm), 100.00% (254nm)

[1507] MS(ESI):C 95 H 134 N 16 O 30 Calculated mass 1978.95, observed m / z 1979.9641 [M+H] + .

[1508] Synthesis Example 2

[1509] Preparation of Compound I-13

[1510]

[1511] To a solution of intermediate 8b (14 mg, 5.65 μmol, 1.0 eq) in DCM (0.5 mL) was added TFA (770 mg, 6.75 mmol, 0.5 mL, 1196 eq). The mixture was stirred at 25 °C for 1 hour. The mixture was concentrated in vacuo. The residue was purified by preparative HPLC (column: C 18-4150·30mm·5μm; mobile phase: [A: 0.1M water (TFA) B: acetonitrile]; B%: 15%-45%, 20 minutes) to give the desired product (2 mg, 8.45e-1 μmol, 14.96% yield) as a white solid.

[1512] 1 H NMR (400 MHz, DMSO-d 6 )δ9.71(s,1H),8.99(s,1H),8.74(s,1H),8.40-8.32(m,1H),8.10-7.92(m,1H),7.84-7.75(m,2H) ,7.65-7.53(m,4H),7.43-7.29(m,3H),7.21(d,J=8.4Hz,2H),7.11(s,2H),6.47-6.37(m,2H),5.75 5.61(m,2H),4.90(s,2H),4.87-4.70(m,3H),4.69-4.59(m,1H),4.49-4.40(m,1H),4.39-4.11(m,8H),3.94(d ,J=6.0Hz,2H),3.87-3.80(m,1H),3.78-3.69(m,3H),3.65-3.62(m,2H),3.56-3.49(m,44H),3.47-3.43(m,2H) ,3.27-3.22(m,2H),2.68-2.66(m,2H),2.60(s,3H),2.40-2.36(m,2H),2.34-2.31(m,3H),2.30-2.24(m,3H),2 .04-1.97(m,3H),1.87-1.74(m,3H),1.70-1.60(m,1H),1.49-1.39(m,2H),1.27(s,3H),0.85(t,J=7.2Hz,6H).

[1513] MS(ESI):C 98 H 136 F 2 N 20 O 38 P 2 S 2 Calculated mass 2364.82, observed m / z 1184.1 [M / 2+H] + . HPLC: 96.65% (220nm), 95.84% (254nm).

[1514] Synthesis Example 3

[1515] Preparation of Compound I-49

[1516]

[1517] To N-[[2-[(1S,2S,4R,6R,8S,9S,11S,12R,13S,19S)-12,19-difluoro-6-(3-fluorophenyl)-11-hydroxy-9,13-dimethyl-16-oxo-5,7-dioxolato[10.8.0.02,9.04,8.013,18]eicosane-14,17-dien-8-yl]-2-oxo-ethoxy]methyl]-N-ethyl-carbamic acid [4-[[(2S)-2-[[(2S)-2-amino-3-methyl-butanoyl]amino]propanoyl]amino]phenyl]methyl ester (94.3 mg, 51.4 μmol, 55% purity, 1.0 equiv, TFA) in DMF (0.7 mM) at 0°C was added 4% paraformaldehyde (pH 7.0). L) was added DIEA (19.9 mg, 154 μmol, 26.8 μL, 3.0 equiv) and (2S)-2-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[5-(2,5-dioxopyrrol-1-yl)-2-[3-oxo-3-(2,3,5,6-tetrafluorophenoxy)propyl]benzoyl]amino]ethylamino]acetyl]amino]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]propionylamino]pentanedioic acid (Intermediate 6a', 78.0 mg, 56.5 μmol, 1.1 equiv, TFA). The mixture was stirred at 25° C. for 1 hour. The reaction mixture was quenched with HCOOH until pH=about 7. The reaction mixture was purified by preparative HPLC (column: Phenomenex Luna C 18200×40mm×10μm; mobile phase: [water (FA)-ACN]; B%: 35%-65%, 8 minutes) purification. (2S)-2-[3-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[...

Claims

1. A compound having the following structure (I): in: X 1 , X 2 , X 3 , X 4 and X 5 One of them is CL 1 -R 1 , X 1 , X 2 , X 3 , X 4 and X 5 The other one is CL 2 -R 2 , and X 1 , X 2 , X 3 , X 4 and X 5 The other three are N, CR, 3 or CL 3 -R 3a ; R 1 , R 2 and R 3a Each independently comprises one or more moieties selected from the group consisting of amino acid elements, charged elements, heteroalkylene elements, hydrophilic elements, trigger elements, sacrificial elements, polar caps, payloads, and combinations thereof; provided that R 1 and R 2 At least one of the comprises a payload; R 3 and independently at each occurrence selected from the group consisting of hydrogen, deuterium, alkyl, haloalkyl, halo, alkoxy, haloalkoxy, amino, amine, amido, aldehyde, hydroxyl, cyano, nitro, thiol, carboxyl, carboxyalkyl, alkyl-S(O)3H, alkyl-OP(O)3H, alkyl-P(O)3H, -O-carboxyalkyl, -O-alkyl-S(O)3H, -O-alkyl-OP(O)3H, -O-alkyl-P(O)3H (O)3H, -S(O)3H, -OP(O)3H, -P(O)3H, alkyl-OP(O)3-alkyl, alkyl-P(O)3-alkyl, -O-alkyl-S(O)3-alkyl, -O-alkyl-OP(O)3-alkyl, -O-alkyl-P(O)3-alkyl, -S(O)3-alkyl, -OP(O)3-alkyl, -P(O)3-alkyl, sulfamides, sulfenamides, and carbohydrates; R 4a and R 4b are independently hydrogen, deuterium, halogenated or -SR 4c , where R 4c is a substituted or unsubstituted C6-C 10 aryl or substituted or unsubstituted 5-12 membered heteroaryl; and L 1 , L 2 and L 3 each independently is a direct bond or a linker comprising an optionally substituted alkylene, an optionally substituted alkenylene, an optionally substituted alkynylene, an optionally substituted heteroalkylene, an optionally substituted heteroalkenylene, an optionally substituted heteroalkynylene, a heteroatom linker, an optionally substituted cycloalkylene, an optionally substituted arylene, an optionally substituted heterocyclylene, an optionally substituted heteroarylene, or a combination thereof; as a stereoisomer, enantiomer or tautomer thereof, or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

2. The compound according to claim 1, wherein R 1 Has the following structure: in: L 1a It is an amino acid element; L 1b It is a live component; L 1c is a heteroalkylene element; L 1d It is a hydrophilic element; L 1e is a triggering element; and L 1f It is a sacrificial component; Where L 1a , L 1b , L 1c , L 1d , L 1e and L 1f When present in one or more instances, optionally with L 1a , L 1b , L 1c , L 1d , L 1e and L 1f One or more of the other of the are joined to form one or more rings; n1, n2, n3, n4, n5 and n6 are independently an integer from 0 to 3 at each occurrence, provided that n1+n2+n3+n4+n5+n6=1 or greater; n7 is 1, 2, 3, 4, 5 or 6; and R 1a Yes and L 1a , L 1b , L 1c , L 1d , L 1e or L 1f a covalently bound payload in one occurrence, and said payload is optionally replaced by a polar cap; As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

3. A compound according to any one of claims 1 to 2, wherein R 1 Has the following structure: in: L 1a It is an amino acid element; L 1b It is a live component; L 1c is a heteroalkylene element; L 1d It is a hydrophilic element; L 1e is a triggering element; and L 1f It is a sacrificial component; Where L 1a , L 1b , L 1c , L 1d , L 1e and L 1f When present in one or more instances, optionally with L 1a , L 1b , L 1c , L 1d , L 1e and L 1f One or more of the other of the are joined to form one or more rings; n1, n2, n3, n4, n5 and n6 are independently an integer from 0 to 3 at each occurrence, provided that n1+n2+n3+n4+n5+n6=1 or greater; n7 is 1, 2, 3, 4, 5 or 6; and R 1a is the payload optionally replaced by the polar cap; As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

4. A compound according to any one of claims 1 to 3, wherein R 2 Has the following structure: in: L 2a It is an amino acid element; L 2b It is a live component; L 2c is a heteroalkylene element; L 2d It is a hydrophilic element; L 2e It is the trigger element; m1, m2, m3, m4 and m5 are independently an integer from 0 to 3 at each occurrence, provided that m1+m2+m3+m4+m5=1 or greater; m6 is 1, 2, 3, 4, or 5; and R 2a is hydrogen, alkyl, payload or polar cap, As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

5. A compound according to any one of claims 1 to 4, wherein R 3a Has the following structure: in: L 3a It is an amino acid element; L 3b It is a live component; L 3c is a heteroalkylene element; L 3d It is a hydrophilic element; L 3e It is the trigger element; p1, p2, p3, p4 and p5 are independently an integer from 0 to 3 at each occurrence, provided that p1+p2+p3+p4+p5=1 or greater; p6 is 1, 2, 3, 4, or 5; and R 3b is hydrogen, alkyl or polar cap, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

6. A compound according to any one of claims 2 to 5, wherein: a) n7 is 1, and each of n1 to n6 is 1; b) n7 is 1, and each of n1 to n4 is 0, n5 is 1, and n6 is 1; or c) n7 is 1, and n1 is 0, n2 is 0, n3 is 1, n4 is 0, n5 is 1, and n6 is 1, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

7. A compound according to any one of claims 4 to 6, wherein: a) m6 is 1, and each of m1 to m5 is 1; or b) m6 is 1, m1 is 1, m2 is 0, m3 is 0, m4 is 1, and m5 is 0, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

8. A compound according to any one of claims 5 to 7, wherein: a) p6 is 1, and each of p1 to p5 is 1; or b) p6 is 1, p1 is 1, p2 is 0, p3 is 0, p4 is 1, and p5 is 0, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

9. The compound according to any one of claims 1 to 8, wherein the amino acid element comprises: a) one or more amino acids selected from the group consisting of glycine, alanine, serine, threonine, cysteine, valine, leucine, isoleucine, methionine, proline, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, asparagine, glutamine, histidine, lysine, arginine, sarcosine and β-alanine; or b) one or more amino acids selected from the group consisting of glycine, sarcosine, β-alanine and glutamic acid, as stereoisomers, enantiomers or tautomers thereof, or mixtures thereof; or pharmaceutically acceptable salts, solvates or prodrugs thereof.

10. A compound according to any one of claims 1 to 9, wherein the amino acid element comprises a dipeptide, tripeptide, tetrapeptide or pentapeptide, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

11. The compound according to any one of claims 1 to 10, wherein the amino acid element has one of the following structures: in: R 5a is independently at each occurrence hydrogen, alkyl, hydroxyalkyl or alkoxyalkyl, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

12. A compound according to any one of claims 1 to 11, wherein the charged element comprises a moiety having a negative charge at pH 7.4, as a stereoisomer, enantiomer or tautomer thereof, or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

13. A compound according to any one of claims 1 to 12, wherein the charged element comprises a moiety having a positive charge at pH 7.4, as a stereoisomer, enantiomer or tautomer thereof, or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

14. A compound according to any one of claims 1 to 13, wherein the charged element comprises one or more charged amino acids, one or more carboxylic acids, one or more sulfonic acids, one or more sulfonamides, one or more sulfates, one or more phosphates, one or more quaternary amines, one or more sulfamides, one or more sulfenamides or combinations thereof, as stereoisomers, enantiomers or tautomers thereof or mixtures thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

15. The compound of claim 14, wherein the charged amino acid is aspartic acid, glutamic acid, histidine, lysine or arginine, as a stereoisomer, enantiomer or tautomer thereof, or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

16. The compound of any one of claims 1 to 15, wherein the heteroalkylene element comprises polyethylene glycol or polypropylene glycol, as a stereoisomer, enantiomer or tautomer thereof, or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

17. The compound according to any one of claims 1 to 16, wherein the heteroalkylene element comprises one of the following structures: in: R 5b , R 5c , R 5d and R 5e is independently selected at each occurrence from the group consisting of hydrogen, deuterium, alkyl, haloalkyl, halo, alkoxy, haloalkoxy, amino, hydroxy, cyano, nitro, thiol, carboxyalkyl, alkyl-S(O)3H, alkyl-OP(O)3H, alkyl-P(O)3H, -O-carboxyalkyl, -O-alkyl-S(O)3H, -O-alkyl-OP(O)3H, -O-alkyl-P(O)3H, -S(O)3H, -OP(O)3H, and -P(O)3H, q1 is independently an integer from 1 to 24 at each occurrence, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

18. A compound according to any one of claims 1 to 17, wherein R 1 , R 2 or R 3a Non-cleavable linkers are included as stereoisomers, enantiomers or tautomers or mixtures thereof; or pharmaceutically acceptable salts, solvates or prodrugs thereof.

19. A compound according to any one of claims 1 to 18, wherein R 1 , R 2 or R 3a Includes one of the following structures: in: R 5b , R 5c , R 5d and R 5e is independently selected at each occurrence from the group consisting of hydrogen, deuterium, alkyl, haloalkyl, halo, alkoxy, haloalkoxy, amino, hydroxy, cyano, nitro, thiol, carboxyalkyl, alkyl-S(O)3H, alkyl-OP(O)3H, alkyl-P(O)3H, -O-carboxyalkyl, -O-alkyl-S(O)3H, -O-alkyl-OP(O)3H, -O-alkyl-P(O)3H, -S(O)3H, -OP(O)3H, -P(O)3H, alkyl-OP(O)3-alkyl, alkyl-P(O)3-alkyl, -O-alkyl-S(O)3-alkyl, -O-alkyl-OP(O)3-alkyl, -O-alkyl-P(O)3-alkyl, -S(O)3-alkyl, -OP(O)3-alkyl, -P(O)3-alkyl, sulfamide, sulfenamide; R 5f is independently at each occurrence hydrogen, alkyl, hydroxyalkyl or alkoxyalkyl; R 9 is independently at each occurrence hydrogen or alkyl; q2 is independently an integer from 1 to 25 at each occurrence; and q3 is independently an integer from 5 to 15 at each occurrence, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

20. The compound according to any one of claims 1 to 19, wherein the hydrophilic element comprises polyethylene glycol, polysarcosine, cyclodextrin, c-glycoside or a combination thereof, as a stereoisomer, enantiomer or tautomer thereof or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

21. The compound according to any one of claims 1 to 20, wherein the hydrophilic element comprises one of the following structures: in: R 5b , R 5c , R 5d and R 5e is independently selected at each occurrence from the group consisting of hydrogen, deuterium, alkyl, haloalkyl, halo, alkoxy, haloalkoxy, amino, hydroxy, cyano, nitro, thiol, carboxyalkyl, alkyl-S(O)3H, alkyl-OP(O)3H, alkyl-P(O)3H, -O-carboxyalkyl, -O-alkyl-S(O)3H, -O-alkyl-OP(O)3H, -O-alkyl-P(O)3H, -S(O)3H, -OP(O)3H, and -P(O)3H, R 5g is independently at each occurrence hydrogen, alkyl, hydroxyalkyl, or alkoxyalkyl; and q4 is independently an integer from 1 to 24 at each occurrence, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

22. The compound according to any one of claims 1 to 21, wherein the trigger element comprises: a) a dipeptide, a tripeptide, a tetrapeptide, a pentapeptide, a glucuronide, a disulfide, a phosphate, a diphosphate, a triphosphate, a hydrazone or a combination thereof; b) β-glucuronic acid; c) a dipeptide, a tripeptide, a tetrapeptide or a pentapeptide; d) two or more amino acids selected from the group consisting of valine, citrulline, alanine, glycine, phenylalanine, lysine, or a combination thereof; e) a sequence of amino acids selected from the group consisting of valine-citrulline, valine-alanine, glycine-glycine-phenylalanine-glycine, and combinations thereof; f) One of the following structures, including their combinations: As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

23. The composition according to any one of claims 1 to 22, wherein the sacrificial element comprises: a) p-aminobenzyloxycarbonyl, aminal, hydrazine, disulfide, amide, ester, hydrazine, phosphotriester, diester, β-glucuronide, double bond, triple bond, ether bond, ketone, diol, cyano, nitro, quaternary amine or a combination thereof; b) p-methoxybenzyl, dialkyldialkoxysilane, diaryldialkoxysilane, orthoester, acetal, optionally substituted β-thiopropionate, ketal, phosphoramidate, hydrazone, vinyl ether, imine, aconityl, trityl, polyketal, bisarylhydrazone, diazobenzene, vicinal diol, pyrophosphate diester or a combination thereof, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof; c) One of the following structures: d) The following structure: in: R 6a , R 6b , R 6c and R 6d is independently hydrogen, optionally substituted alkyl, optionally substituted aryl or optionally substituted heteroaryl, or R 6a and R 6c together with the nitrogen and carbon atoms to which they are attached form an azetidinyl, pyrrolidinyl, piperidinyl or homopiperidinyl group, and R 6d is hydrogen; and Y 1 Is –O–, –S–, or –NR 6b – e) The following structure: in: R 6e , R 6f , R 6g and R 6h is independently hydrogen, optionally substituted alkyl, optionally substituted aryl or optionally substituted heteroaryl, or R 6a and R 6c together with the nitrogen and carbon atoms to which they are attached form an azetidinyl, pyrrolidinyl, piperidinyl or homopiperidinyl group, and R 6d is hydrogen; and Y 2 Is –O–, –S–, or –NR 6f – f) The following structure: in: R 10 is independently at each occurrence alkyl, alkoxy or halo; R 11 is hydrogen, alkyl or –(CH2CH2O) z3 -CH3; R 12 is hydrogen or alkyl; R 13 is hydrogen or alkyl; z1 is 0 or 1; z2 is 0, 1, 2, 3 or 4; and z3 is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; or g) One of the following structures: in: R 14a , R 14b , R 14c , R 14d , R 14e and R 14f are each independently hydrogen, alkyl, hydroxyalkyl or alkoxyalkyl; z4, z5, z6 and z7 are each independently 1, 2, 3, 4, 5 or 6; h) One of the following structures: in: z8 and z9 are each independently 1, 2, 3, 4, 5 or 6; or i) One of the following structures: in: R 15 is independently at each occurrence H, methyl, ethyl, isopropyl, tert-butyl or phenyl; Y 3 is O or CH2; and q5 is an integer in the range of 1-5, as a stereoisomer, enantiomer or tautomer thereof, or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

24. A compound according to any one of claims 1 to 23, wherein R 4a and R 4b are all hydrogen, as stereoisomers, enantiomers or tautomers or mixtures thereof; or pharmaceutically acceptable salts, solvates or prodrugs thereof.

25. The compound according to any one of claims 1 to 24, wherein the trigger element and the sacrificial element together comprise one of the following structures: As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

26. The compound of any one of claims 1 to 25, wherein the polar cap comprises: a) one or more charged amino acids, one or more polyols, or a combination thereof; b) diols, triols, tetraols or combinations thereof; c) glycerol, trimethylolpropane, pentaerythritol, maltitol, sorbitol, xylitol, erythritol, isomalt or a combination thereof; d) one or more naturally occurring amino acids; e) one or more unnatural amino acids; f) one or more unnatural amino acids and one or more natural amino acids; g) serine, threonine, cysteine, proline, asparagine, glutamine, lysine, arginine, histidine, aspartate, glutamate, 4-hydroxyproline, 5-hydroxylysine, homoserine, homocysteine, ornithine, β-alanine, statine or γ-aminobutyric acid; or h) aspartic acid, serine, glutamic acid, serine-β-glucose or a combination thereof, as a stereoisomer, enantiomer or tautomer thereof or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

27. The compound of any one of claims 1 to 26, wherein the polar cap has one of the following structures, including combinations thereof: As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

28. A compound according to any one of claims 1 to 27, wherein the payload is a chemotherapeutic agent, a cytotoxic agent or a myeloid cell agonist, as a stereoisomer, enantiomer or tautomer thereof, or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

29. The compound according to any one of claims 1 to 28, wherein the payload is: a) a chemotherapeutic agent selected from the group consisting of camptothecin, paclitaxel, doxorubicin, vinblastine, dacarbazine, irinotecan, topotecan, silatecan, cositecan, exatecan, lurtotecan, SN-38, Dxd, gimatecan, Belotecan and rubitecan; b) a cytotoxic agent selected from the group consisting of calicheamicin, anthramycin, abbeymycin, chicamycin, DC-81, mazethramycin, neothramycin A, neothramycin B, porothramycin, prothracarcin, sibanomicin, sibiromycin, tomamycin, auristatin F, monomethyl auristatin F, auristatin E, monomethyl auristatin E, dolastatin, monomethyl dolastatin, mertansine, and emtansine; or c) a myeloid cell agonist selected from the group consisting of a STING agonist, a ligand of TLR2, a ligand of TLR3, a ligand of TLR4, a ligand of TLR5, a ligand of TLR6, a ligand of TLR7, a ligand of TLR8, a ligand of TLR9, a ligand of TLR10, a ligand of a nucleotide oligomerization domain (NOD), a ligand of a RIG-I-like receptor (RLR), a ligand of a C-type lectin receptor (CLR), a ligand of a cytoplasmic DNA sensor (CDS), and a ligand of an inflammasome inducer, preferably wherein the myeloid cell agonist is selected from the group consisting of a Sergan selgantolimod, motolimod, resiquimod, 3M-051, 3M-052, MCT-465, IMO-4200, VTX-763, VTX-1463, RG7854, ADU-S100, MK-1454, MK-2118, BMS-986301, GSK3745417, SB-11285 and IMSA-101, as stereoisomers, enantiomers or tautomers thereof or mixtures thereof; or pharmaceutically acceptable salts, solvates or prodrugs thereof.

30. A compound according to any one of claims 1 to 29, wherein L 1 , L 2 or L 3 A linker comprising a linker selected from the group consisting of: alkylene, alkylene-L a -, alkenylene, alkenylene-L a -, alkynylene, alkynylene-L a -、-L a -、-L a -Alkylene-L a -、-L a -Alkenylene-L a -、-L a -Alkynylidene-L a - and combinations thereof, wherein each alkylene, alkenylene and alkynylene is optionally substituted; L a is independently selected at each occurrence from -O-, -S-, -N(R 7 )-, -C(O)-, -C(S)-, -C(O)O-, -OC(O)-, -OC(O)O-, -C(O)N(R 7 )-、-N(R 7 )C(O)-、-C(O)N(R 7 )C(O)-、 -C(O)N(R 7 )C(O)N(R 7 )、-N(R 7 )C(O)N(R 7 )-、-N(R 7 )C(O)O-、-OC(O)N(R 7 )-、-C(NR 7 )-、-N(R 7 )C(NR 7 )-、-C(NR 7 )N(R 7 )-、-N(R 7 )C(NR 7 )N(R 7 )-、-S(O)2-、-OS(O)-、-S(O)O-、-S(O)、-OS(O)2-、-S(O)2O、-N(R 7 )S(O)2-、-S(O)2N(R 7 )-、-N(R 7 )S(O)-、-S(O)N(R 7 )-、-N(R 7 )S(O)2N(R 7 )- and -N(R 7 )S(O)N(R 7 )-; R 7 is independently selected at each occurrence from hydrogen, -NH2, -C(O)OCH2C6H5; and C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-12 Cycloalkyl and 3 to 12 membered heterocyclic rings, each of which is optionally substituted by one or more substituents independently selected from the group consisting of halo, hydroxy, cyano, nitro, amino, oxo, thioxo, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 Alkyl, C 1-10 Haloalkyl, C 1-10 Alkoxy, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-12 Cycloalkyl and 3 to 12 membered heterocyclyl, as stereoisomers, enantiomers or tautomers or mixtures thereof; or pharmaceutically acceptable salts, solvates or prodrugs thereof.

31. A compound according to any one of claims 1 to 30, wherein each L 1 , L 2 or L 3 Optionally substituted with one or more substituents selected from the group consisting of alkyl, alkenyl, alkynyl, halo, hydroxy, cyano, -OR 8 、-SR 8 , amino, amine, acylamino, cycloalkyl, aryl, heterocyclyl, heteroaryl, cycloalkylalkyl, arylalkyl, heterocycloalkylalkyl, heteroarylalkyl, -C(O)R 8 、-C(O)N(R 8 )2、-N(R 8 )C(O)R 8 、-C(O)OR 8 、-OC(O)R 8 、-S(O)R 8 、-S(O)2R 8 、-P(O)(OR 8 )2.-OP(O)(OR 8 )2, nitro, oxo, thio, =N(R 8 ) or cyano; and R 8 is hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heterocyclyl, heteroaryl, cycloalkylalkyl, arylalkyl, heterocycloalkylalkyl or heteroarylalkyl, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

32. A compound according to any one of claims 1 to 31, wherein L 1 , L 2 or L 3 Independently selected from the following structures: in: R a is hydrogen or alkyl; L b is independently at each occurrence a direct bond, an optionally substituted alkylene linker, an optionally substituted heteroalkylene linker, a heteroatom linker, or a combination thereof; L c is independently at each occurrence an optionally substituted alkylene linker; The condition is L 1 , L 2 or L 3 At least one of has the following structure; As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

33. A compound according to any one of claims 1 to 32, wherein L 1 and L 2 Independently selected from the following structures: in: R a is hydrogen or alkyl; L b is independently at each occurrence a direct bond, an optionally substituted alkylene linker, an optionally substituted heteroalkylene linker, a heteroatom linker, or a combination thereof; L c is independently at each occurrence an optionally substituted alkylene linker; The condition is L 1 or L 2 At least one of has the following structure; As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

34. A compound according to any one of claims 1 to 33, wherein L 2 Has the following structure: As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

35. A compound according to any one of claims 1 to 34, wherein L 1 , L 2 and L 3 Each independently has at least one of the following structures: in: *Indicates a direct bond to a substitutable position on the phenyl group of the following structure: As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

36. A compound according to any one of claims 1 to 35, wherein L 1 or L 2 Has one of the following structures: in: *Indicates a direct bond to a substitutable position on the phenyl group of the following structure: As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

37. A compound according to any one of claims 1 to 36, wherein L c is a C1-C6 alkylene group, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

38. A compound according to any one of claims 1 to 37, wherein L c Substituted with one or more substituents selected from the group consisting of halo, haloalkyl, alkoxy, cyano, nitro, carboxyl, sulfonamide, sulfonic acid or a combination thereof, as stereoisomers, enantiomers or tautomers or mixtures thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

39. A compound according to any one of claims 1 to 38, wherein the compound has one of the following structures (Ia) or (Ib): As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

40. A compound according to any one of claims 1 to 38, wherein the compound has one of the following structures (Ic') or (Ic"): As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

41. A compound according to any one of claims 1 to 39, wherein X 2 , X 3 Or both are CH or CF.

42. A compound according to any one of claims 1 to 41, wherein X 1 , X 5 Or both are CR 3 , and R 3 is H or halo, preferably wherein halo is F.

43. A compound according to any one of claims 1 to 42, wherein X 1 , X 5 Or both are CR 3 , and R 3 is halo, preferably wherein halo is F.

44. A compound according to any one of claims 1 to 42, wherein X 1 is CH, and X 5 It is CH.

45. A compound according to any one of claims 1 to 43, wherein X 1 is CF, and X 5 It is CH.

46. ​​A compound according to any one of claims 1 to 43, wherein X 1 is CH, and X 5 It's CF.

47. A compound according to any one of claims 1 to 38, wherein the compound has one of the following structures: (Ia-1), (Ia-2), (Ia-3), (Ia-4), (Ia-5), (Ia-6), (Ia-7) or (Ia-8): in: L b is independently at each occurrence a direct bond, an optionally substituted alkylene linker, an optionally substituted heteroalkylene linker, a heteroatom linker, or a combination thereof; and q6 is 0, 1, or 2, As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

48. The compound according to any one of claims 1 to 38, wherein the compound has one of the following structures: (Ic-1), (Ic-2), (Ic-3) or (Ic-4): in: L b is independently at each occurrence a direct bond, an optionally substituted alkylene linker, an optionally substituted heteroalkylene linker, a heteroatom linker, or a combination thereof; and q7 is 1, 2 or 3, as a stereoisomer, enantiomer or tautomer thereof or a mixture thereof; or A pharmaceutically acceptable salt, solvate or prodrug thereof.

49. A compound according to any one of claims 1 to 38, wherein the compound has the following structure: (Id), (Ie), (If) or (Ig): in: L b is independently at each occurrence a direct bond, an optionally substituted alkylene linker, an optionally substituted heteroalkylene linker, a heteroatom linker, or a combination thereof; q8 is 0, 1, or 2; and q9 is 0, 1 or 2, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or A pharmaceutically acceptable salt, solvate or prodrug thereof.

50. A compound according to any one of claims 1 to 38, wherein the compound has one of the following structures (Ih) or (Ii): in: L b is independently at each occurrence a direct bond, an optionally substituted alkylene linker, an optionally substituted heteroalkylene linker, a heteroatom linker, or a combination thereof, as a stereoisomer, enantiomer or tautomer, or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

51. according to the compound described in any one of claims 32 to 50, wherein L b is a direct bond, an optionally substituted alkylene linker or an optionally substituted heteroalkylene linker, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

52. according to the compound described in any one of claims 32 to 51, wherein L b is a direct key or has one of the following structures: in: R b is independently at each occurrence hydrogen, alkyl, hydroxyalkyl or alkoxyalkyl, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

53. A compound according to any one of claims 1 to 38, wherein the compound has one of the following structures: in: R 1b is a chemotherapeutic, cytotoxic, or myeloid cell agonist; R 2b Has one of the following structures: L 1g Has one of the following structures: As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

54. A compound according to any one of claims 2 to 53, wherein the compound has one of the following structures: As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

55. according to the compound described in any one of claims 2 to 54, wherein R 1a Has one of the following structures: As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

56. according to the compound described in any one of claims 2 to 54, wherein R 1a Has one of the following structures: in: R' is hydrogen or has one of the following structures: in: R a ' is H or C 1-6 alkyl; R b 'It's C 1-6 Alkyl or C 1-6 Alkoxy; R c 'It is H, C 1-6 Alkyl, -CH2OH or C 1-6 Alkoxy; R d ' is H or C 1-6 Alkyl; or R e ' is H or C 1-6 alkyl.

57. A compound according to any one of claims 1 to 56, wherein L 1 or L 2 Has the following structure: in: ** Shown with X 1 , X 2 , X 3 , X 4 or X 5 a bond of , as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

58. A compound according to any one of claims 1 to 31, wherein X 2 It is CL 1 -R 1 , and X 3 It is CL 2 -R 2 , as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

59. A compound according to any one of claims 1 to 31, wherein X 3 It is CL 1 -R 1 , and X 2 It is CL 2 -R 2 , as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

60. A compound having a structure selected from Table 1 as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

61. A conjugate having the following structure (II): in: A is a targeting moiety or a binding fragment thereof; L 4 Has one of the following structures: *** indicates the connection point with A; g is an integer from 1 to 20; X 1 , X 2 , X 3 , X 4 and X 5 One of them is CL 1 -R 1 , X 1 , X 2 , X 3 , X 4 and X 5 The other one is CL 2 -R 2 , and X 1 , X 2 , X 3 , X 4 and X 5 The other three are N, CR, 3 or CL 3 -R 3a ; R 1 , R 2 and R 3a Each independently comprises one or more moieties selected from the group consisting of amino acid elements, charged elements, heteroalkylene elements, hydrophilic elements, trigger elements, sacrificial elements, polar caps, payloads, and combinations thereof; The condition is R 1 and R 2 At least one of the comprises a payload; R 3 and independently at each occurrence selected from the group consisting of hydrogen, deuterium, alkyl, haloalkyl, halo, alkoxy, haloalkoxy, amino, amine, amido, aldehyde, hydroxyl, cyano, nitro, thiol, carboxyl, carboxyalkyl, alkyl-S(O)3H, alkyl-OP(O)3H, alkyl-P(O)3H, -O-carboxyalkyl, -O-alkyl-S(O)3H, -O-alkyl-OP(O)3H, -O-alkyl-P(O)3H (O)3H, -S(O)3H, -OP(O)3H, -P(O)3H, alkyl-OP(O)3-alkyl, alkyl-P(O)3-alkyl, -O-alkyl-S(O)3-alkyl, -O-alkyl-OP(O)3-alkyl, -O-alkyl-P(O)3-alkyl, -S(O)3-alkyl, -OP(O)3-alkyl, -P(O)3-alkyl, sulfamides, sulfenamides, and carbohydrates; R 4a is hydrogen, deuterium, halogenated or -SR 4c , where R 4c is a substituted or unsubstituted C6-C 10 aryl or substituted or unsubstituted 5-12 membered heteroaryl; and L 1 , L 2 and L 3 each is independently a direct bond or a linker comprising an optionally substituted alkylene, an optionally substituted alkenylene, an optionally substituted alkynylene, an optionally substituted heteroalkylene, an optionally substituted heteroalkenylene, an optionally substituted heteroalkynylene, a heteroatom linker, an optionally substituted cycloalkylene, an optionally substituted arylene, an optionally substituted heterocyclylene, an optionally substituted heteroarylene, or a combination thereof, as a stereoisomer, enantiomer or tautomer thereof, or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

62. The conjugate according to claim 61, wherein R 1 Has the following structure: in: L 1a It is an amino acid element; L 1b It is a live component; L 1c is a heteroalkylene element; L 1d It is a hydrophilic element; L 1e is a triggering element; and L 1f It is a sacrificial component; Where L 1a , L 1b , L 1c , L 1d , L 1e and L 1f When present in one or more instances, optionally with L 1a , L 1b , L 1c , L 1d , L 1e and L 1f One or more of the other of the are joined to form one or more rings; n1, n2, n3, n4, n5 and n6 are independently an integer from 0 to 3 at each occurrence, provided that n1+n2+n3+n4+n5+n6=1 or greater; n7 is 1, 2, 3, 4, 5 or 6; and R 1a Yes and L 1a , L 1b , L 1c , L 1d , L 1e or L 1f A covalently bound payload in one occurrence, and the payload is optionally substituted with a polar cap, as a stereoisomer, enantiomer or tautomer, or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

63. The conjugate according to any one of claims 61 to 62, wherein R 1 Has the following structure: in: L 1a It is an amino acid element; L 1b It is a live component; L 1c is a heteroalkylene element; L 1d It is a hydrophilic element; L 1e is a triggering element; and L 1f It is a sacrificial component; Where L 1a , L 1b , L 1c , L 1d , L 1e and L 1f When present in one or more instances, optionally with L 1a , L 1b , L 1c , L 1d , L 1e and L 1f One or more of the other of the are joined to form one or more rings; n1, n2, n3, n4, n5 and n6 are independently an integer from 0 to 3 at each occurrence, provided that n1+n2+n3+n4+n5+n6=1 or greater; n7 is 1, 2, 3, 4, 5 or 6; and R 1a is an optionally polar-cap substituted payload, as a stereoisomer, enantiomer or tautomer, or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

64. The conjugate of any one of claims 61 to 63, wherein R 2 Has the following structure: in: L 2a It is an amino acid element; L 2b It is a live component; L 2c is a heteroalkylene element; L 2d It is a hydrophilic element; L 2e It is the trigger element; m1, m2, m3, m4 and m5 are independently an integer from 0 to 3 at each occurrence, provided that m1+m2+m3+m4+m5=1 or greater; m6 is 1, 2, 3, 4, or 5; and R 2a is hydrogen, alkyl, payload or polar cap, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

65. The conjugate of any one of claims 61 to 64, wherein R 3a Has the following structure: in: L 3a It is an amino acid element; L 3b It is a live component; L 3c is a heteroalkylene element; L 3d It is a hydrophilic element; L 3e It is the trigger element; p1, p2, p3, p4 and p5 are independently an integer from 0 to 3 at each occurrence, provided that p1+p2+p3+p4+p5=1 or greater; p6 is 1, 2, 3, 4, or 5; and R 3b is hydrogen, alkyl or polar cap, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

66. The conjugate according to any one of claims 62 to 65, wherein: a) n7 is 1, and each of n1 to n6 is 1; b) n7 is 1, and each of n1 to n4 is 0, n5 is 1, and n6 is 1; or c) n7 is 1, and n1 is 0, n2 is 0, n3 is 1, n4 is 0, n5 is 1, and n6 is 1, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

67. The conjugate according to any one of claims 64 to 66, wherein: a) m6 is 1, and each of m1 to m5 is 1; or b) m6 is 1, m1 is 1, m2 is 0, m3 is 0, m4 is 1, and m5 is 0, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

68. The conjugate according to any one of claims 65 to 67, wherein: a) p6 is 1, and each of p1 to p5 is 1; or b) p6 is 1, p1 is 1, p2 is 0, p3 is 0, p4 is 1, and p5 is 0, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

69. The conjugate of any one of claims 61 to 67, wherein the amino acid element comprises: a) one or more amino acids selected from the group consisting of glycine, alanine, serine, threonine, cysteine, valine, leucine, isoleucine, methionine, proline, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, asparagine, glutamine, histidine, lysine, arginine, sarcosine and β-alanine; or b) one or more amino acids selected from the group consisting of glycine, sarcosine, β-alanine and glutamic acid, as stereoisomers, enantiomers or tautomers thereof, or mixtures thereof; or pharmaceutically acceptable salts, solvates or prodrugs thereof.

70. The conjugate of any one of claims 61 to 69, wherein the amino acid element comprises a dipeptide, tripeptide, tetrapeptide or pentapeptide, as a stereoisomer, enantiomer or tautomer thereof, or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

71. The conjugate of any one of claims 61 to 70, wherein the amino acid element has one of the following structures: in: R 5a is independently at each occurrence hydrogen, alkyl, hydroxyalkyl or alkoxyalkyl, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

72. The conjugate of any one of claims 61 to 71, wherein the charged element comprises a moiety having a negative charge at pH 7.4, as a stereoisomer, enantiomer or tautomer thereof, or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

73. The conjugate of any one of claims 61 to 72, wherein the charged element comprises a moiety having a positive charge at pH 7.4, as a stereoisomer, enantiomer or tautomer thereof, or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

74. The conjugate of any one of claims 61 to 73, wherein the charged element comprises one or more charged amino acids, one or more carboxylic acids, one or more sulfonic acids, one or more sulfonamides, one or more sulfates, one or more phosphates, one or more quaternary amines, one or more sulfamides, one or more sulfenamides, or a combination thereof, as stereoisomers, enantiomers or tautomers thereof, or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

75. The conjugate of claim 74, wherein the charged amino acid is aspartic acid, glutamic acid, histidine, lysine or arginine, as a stereoisomer, enantiomer or tautomer thereof, or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

76. The conjugate of any one of claims 61 to 75, wherein the heteroalkylene element comprises polyethylene glycol or polypropylene glycol, as a stereoisomer, enantiomer or tautomer thereof, or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

77. The conjugate of any one of claims 61 to 76, wherein the heteroalkylene element comprises one of the following structures: in: R 5b , R 5c , R 5d and R 5e is independently selected at each occurrence from the group consisting of hydrogen, deuterium, alkyl, haloalkyl, halo, alkoxy, haloalkoxy, amino, hydroxy, cyano, nitro, thiol, carboxyalkyl, alkyl-S(O)3H, alkyl-OP(O)3H, alkyl-P(O)3H, -O-carboxyalkyl, -O-alkyl-S(O)3H, -O-alkyl-OP(O)3H, -O-alkyl-P(O)3H, -S(O)3H, -OP(O)3H, and -P(O)3H, q1 is independently an integer from 1 to 24 at each occurrence, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

78. The conjugate of any one of claims 61 to 77, wherein R 1 , R 2 or R 3a Non-cleavable linkers are included as stereoisomers, enantiomers or tautomers or mixtures thereof; or pharmaceutically acceptable salts, solvates or prodrugs thereof.

79. The conjugate of any one of claims 61 to 78, wherein R 1 , R 2 or R 3a Includes one of the following structures: in: R 5b , R 5c , R 5d and R 5e is independently selected at each occurrence from the group consisting of hydrogen, deuterium, alkyl, haloalkyl, halo, alkoxy, haloalkoxy, amino, hydroxy, cyano, nitro, thiol, carboxyalkyl, alkyl-S(O)3H, alkyl-OP(O)3H, alkyl-P(O)3H, -O-carboxyalkyl, -O-alkyl-S(O)3H, -O-alkyl -OP(O)3H, -O-alkyl-P(O)3H, -S(O)3H, -OP(O)3H, -P(O)3H, alkyl-OP(O)3-alkyl, alkyl-P(O)3-alkyl, -O-alkyl-S(O)3-alkyl, -O-alkyl-OP(O)3-alkyl, -O-alkyl-P(O)3-alkyl, -S(O)3-alkyl, -OP(O)3-alkyl, -P(O)3-alkyl, sulfamide, sulfenamide; R 5f is independently at each occurrence hydrogen, alkyl, hydroxyalkyl or alkoxyalkyl; R 9 is independently at each occurrence hydrogen or alkyl; q2 is independently an integer from 1 to 25 at each occurrence; and q3 is independently an integer from 5 to 15 at each occurrence, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

80. The conjugate of any one of claims 61 to 79, wherein the hydrophilic element comprises polyethylene glycol, polysarcosine, cyclodextrin, c-glycoside, or a combination thereof, as a stereoisomer, enantiomer, or tautomer, or a mixture thereof; or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

81. The conjugate of any one of claims 61 to 80, wherein the hydrophilic element comprises one of the following structures: in: R 5b , R 5c , R 5d and R 5e is independently selected at each occurrence from the group consisting of hydrogen, deuterium, alkyl, haloalkyl, halo, alkoxy, haloalkoxy, amino, hydroxy, cyano, nitro, thiol, carboxyalkyl, alkyl-S(O)3H, alkyl-OP(O)3H, alkyl-P(O)3H, -O-carboxyalkyl, -O-alkyl-S(O)3H, -O-alkyl-OP(O)3H, -O-alkyl-P(O)3H, -S(O)3H, -OP(O)3H, and -P(O)3H, R 5g is independently at each occurrence hydrogen, alkyl, hydroxyalkyl, or alkoxyalkyl; and q4 is independently an integer from 1 to 24 at each occurrence, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

82. The conjugate of any one of claims 61 to 81, wherein the trigger element comprises: a) a dipeptide, a tripeptide, a tetrapeptide, a pentapeptide, a glucuronide, a disulfide, a phosphate, a diphosphate, a triphosphate, a hydrazone or a combination thereof; b) β-glucuronic acid; c) a dipeptide, a tripeptide, a tetrapeptide or a pentapeptide; d) two or more amino acids selected from the group consisting of valine, citrulline, alanine, glycine, phenylalanine, lysine, or a combination thereof; e) a sequence of amino acids selected from the group consisting of valine-citrulline, valine-alanine, glycine-glycine-phenylalanine-glycine, and combinations thereof; f) One of the following structures, including their combinations: As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

83. The conjugate of any one of claims 61 to 82, wherein the sacrificial element comprises: a) p-aminobenzyloxycarbonyl, aminal, hydrazine, disulfide, amide, ester, hydrazine, phosphotriester, diester, β-glucuronide, double bond, triple bond, ether bond, ketone, diol, cyano, nitro, quaternary amine or a combination thereof; b) p-methoxybenzyl, dialkyldialkoxysilane, diaryldialkoxysilane, orthoester, acetal, optionally substituted β-thiopropionate, ketal, phosphoramidate, hydrazone, vinyl ether, imine, aconityl, trityl, polyketal, bisarylhydrazone, diazobenzene, vicinal diol, pyrophosphate diester or a combination thereof, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof; c) One of the following structures: d) The following structure: in: R 6a , R 6b , R 6c and R 6d is independently hydrogen, optionally substituted alkyl, optionally substituted aryl or optionally substituted heteroaryl, or R 6a and R 6c together with the nitrogen and carbon atoms to which they are attached form an azetidinyl, pyrrolidinyl, piperidinyl or homopiperidinyl group, and R 6d is hydrogen; and Y 1 Is –O–, –S–, or –NR 6b – e) The following structure: in: R 6e , R 6f , R 6g and R 6h is independently hydrogen, optionally substituted alkyl, optionally substituted aryl or optionally substituted heteroaryl, or R 6a and R 6c together with the nitrogen and carbon atoms to which they are attached form an azetidinyl, pyrrolidinyl, piperidinyl or homopiperidinyl group, and R 6d is hydrogen; and Y 2 Is –O–, –S–, or –NR 6f – f) The following structure: in: R 10 is independently at each occurrence alkyl, alkoxy or halo; R 11 is hydrogen, alkyl or –(CH2CH2O) z3 -CH3; R 12 is hydrogen or alkyl; R 13 is hydrogen or alkyl; z1 is 0 or 1; z2 is 0, 1, 2, 3 or 4; and z3 is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; or g) One of the following structures: in: R 14a , R 14b , R 14c , R 14d , R 14e and R 14f are each independently hydrogen, alkyl, hydroxyalkyl or alkoxyalkyl; z4, z5, z6 and z7 are each independently 1, 2, 3, 4, 5 or 6; h) One of the following structures: in: z8 and z9 are each independently 1, 2, 3, 4, 5 or 6; or i) One of the following structures: in: R 15 is independently H, methyl, ethyl, isopropyl, tert-butyl or phenyl at each occurrence; Y 3 is O or CH2; and q5 is an integer in the range of 1-5, as a stereoisomer, enantiomer or tautomer thereof, or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

84. The conjugate of any one of claims 61 to 83, wherein R 4a is hydrogen, and ---- is a single bond, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

85. The conjugate of any one of claims 61 to 84, wherein the trigger element and the sacrificial element together comprise one of the following structures: As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

86. The conjugate of any one of claims 61 to 85, wherein the polar cap comprises: a) one or more charged amino acids, one or more polyols, or a combination thereof; b) diols, triols, tetraols or combinations thereof; c) glycerol, trimethylolpropane, pentaerythritol, maltitol, sorbitol, xylitol, erythritol, isomalt or a combination thereof; d) one or more naturally occurring amino acids; e) one or more unnatural amino acids; f) one or more unnatural amino acids and one or more natural amino acids; g) serine, threonine, cysteine, proline, asparagine, glutamine, lysine, arginine, histidine, aspartate, glutamate, 4-hydroxyproline, 5-hydroxylysine, homoserine, homocysteine, ornithine, β-alanine, statin or γ-aminobutyric acid; h) aspartic acid, serine, glutamic acid, serine-β-glucose or a combination thereof, as a stereoisomer, enantiomer or tautomer thereof or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

87. The conjugate of any one of claims 61 to 86, wherein the polar cap has one of the following structures, including combinations thereof: As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

88. The conjugate of any one of claims 61 to 87, wherein the payload is a chemotherapeutic agent, a cytotoxic agent, or a myeloid cell agonist, as a stereoisomer, enantiomer, or tautomer, or a mixture thereof; or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

89. The conjugate of any one of claims 61 to 88, wherein the payload is: a) a chemotherapeutic agent selected from the group consisting of camptothecin, paclitaxel, doxorubicin, vinblastine, dacarbazine, irinotecan, topotecan, xilatecan, cositect, exitecan, lortotecan, SN-38, Dxd, gematitecan, belotecan, and rubitecan; b) a cytotoxic agent selected from the group consisting of calicheamicin, anthramycin, abemycin, cheikamycin, DC-81, methylaminomycin, neoaminomycin A, neoaminomycin B, puromycin, ploracomycin, sibamicin, sibirinomycin, tomamycin, auristatin F, monomethyl auristatin F, auristatin E, monomethyl auristatin E, dolestatin, monomethyl dolestatin, maytansine, and maytansine; or c) a myeloid cell agonist selected from the group consisting of a STING agonist, a ligand of TLR2, a ligand of TLR3, a ligand of TLR4, a ligand of TLR5, a ligand of TLR6, a ligand of TLR7, a ligand of TLR8, a ligand of TLR9, a ligand of TLR10, a ligand of a nucleotide oligomerization domain (NOD), a ligand of a RIG-I-like receptor (RLR), a ligand of a C-type lectin receptor (C LR), a ligand of a cytoplasmic DNA sensor (CDS) and a ligand of an inflammasome inducer, preferably wherein the myeloid cell agonist is selected from the group consisting of: sergantomod, motomod, resiquimod, 3M-051, 3M-052, MCT-465, IMO-4200, VTX-763, VTX-1463, RG7854, ADU-S100, MK-1454, MK-2118, BMS-986301, GSK3745417, SB-11285 and IMSA-101, as stereoisomers, enantiomers or tautomers or mixtures thereof; or pharmaceutically acceptable salts, solvates or prodrugs thereof.

90. The conjugate of any one of claims 61 to 89, wherein L 1 , L 2 or L 3 A linker comprising a linker selected from the group consisting of: alkylene, alkylene-L a -, alkenylene, alkenylene-L a -, alkynylene, alkynylene-L a -、-L a -、-L a -Alkylene-L a -、-L a -Alkenylene-L a -、-L a -Alkynylidene-L a - and combinations thereof, wherein each alkylene, alkenylene and alkynylene is optionally substituted; L a is independently selected at each occurrence from -O-, -S-, -N(R 7 )-, -C(O)-, -C(S)-, -C(O)O-, -OC(O)-, -OC(O)O-, -C(O)N(R 7 )-、-N(R 7 )C(O)-、-C(O)N(R 7 )C(O)-、 -C(O)N(R 7 )C(O)N(R 7 )、-N(R 7 )C(O)N(R 7 )-、-N(R 7 )C(O)O-、-OC(O)N(R 7 )-、-C(NR 7 )-、-N(R 7 )C(NR 7 )-、-C(NR 7 )N(R 7 )-、-N(R 7 )C(NR 7 )N(R 7 )-、-S(O)2-、-OS(O)-、-S(O)O-、-S(O)、-OS(O)2-、-S(O)2O、-N(R 7 )S(O)2-、-S(O)2N(R 7 )-、-N(R 7 )S(O)-、-S(O)N(R 7 )-、-N(R 7 )S(O)2N(R 7 )- and -N(R 7 )S(O)N(R 7 )-; R 7 is independently selected at each occurrence from hydrogen, -NH2, -C(O)OCH2C6H5; and C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-12 Cycloalkyl and 3 to 12 membered heterocyclic rings, each of which is optionally substituted by one or more substituents independently selected from the group consisting of halo, hydroxy, cyano, nitro, amino, oxo, thioxo, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 Alkyl, C 1-10 Haloalkyl, C 1-10 Alkoxy, C 2-10 Alkenyl, C 2-10 Alkynyl, C 3-12 Cycloalkyl and 3 to 12 membered heterocyclyl, as stereoisomers, enantiomers or tautomers or mixtures thereof; or pharmaceutically acceptable salts, solvates or prodrugs thereof.

91. The conjugate of any one of claims 61 to 90, wherein each L 1 , L 2 or L 3 Optionally substituted with one or more substituents selected from the group consisting of alkyl, alkenyl, alkynyl, halo, hydroxy, cyano, -OR 8 、-SR 8 , amino, amine, acylamino, cycloalkyl, aryl, heterocyclyl, heteroaryl, cycloalkylalkyl, arylalkyl, heterocycloalkylalkyl, heteroarylalkyl, -C(O)R 8 、-C(O)N(R 8 )2、-N(R 8 )C(O)R 8 、-C(O)OR 8 、-OC(O)R 8 、-S(O)R 8 、-S(O)2R 8 、-P(O)(OR 8 )2.-OP(O)(OR 8 )2, nitro, oxo, thio, =N(R 8 ) or cyano; and R 8 is hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heterocyclyl, heteroaryl, cycloalkylalkyl, arylalkyl, heterocycloalkylalkyl or heteroarylalkyl, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

92. The conjugate of any one of claims 61 to 91, wherein L 1 , L 2 or L 3 Independently selected from the following structures: in: R a is hydrogen or alkyl; L b is independently at each occurrence a direct bond, an optionally substituted alkylene linker, an optionally substituted heteroalkylene linker, a heteroatom linker, or a combination thereof; L c is independently at each occurrence an optionally substituted alkylene linker; The condition is L 1 , L 2 or L 3 At least one of has the following structure; As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

93. The conjugate of any one of claims 61 to 92, wherein L 1 and L 2 Independently selected from the following structures: in: R a is hydrogen or alkyl; L b is independently at each occurrence a direct bond, an optionally substituted alkylene linker, an optionally substituted heteroalkylene linker, a heteroatom linker, or a combination thereof; L c is independently at each occurrence an optionally substituted alkylene linker; provided that L 1 or L 2 At least one of has the following structure; As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

94. The conjugate of any one of claims 61 to 93, wherein L 2 Has the following structure: As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

95. The conjugate of any one of claims 61 to 94, wherein L 1 , L 2 and L 3 Each independently has at least one of the following structures: in: *Indicates a direct bond to a substitutable position on the phenyl group of the following structure: As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

96. The conjugate of any one of claims 61 to 95, wherein L 1 or L 2 Has one of the following structures: in: *Indicates a direct bond to a substitutable position on the phenyl group of the following structure: As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

97. The conjugate of any one of claims 61 to 96, wherein L c is a C1-C6 alkylene group, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

98. The conjugate of any one of claims 61 to 97, wherein L c Substituted with one or more substituents selected from the group consisting of halo, haloalkyl, alkoxy, cyano, nitro, carboxyl, sulfonamide, sulfonic acid or a combination thereof, as stereoisomers, enantiomers or tautomers or mixtures thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

99. The conjugate of any one of claims 61 to 98, wherein the conjugate has one of the following structures (IIa) or (IIb): As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

100. The conjugate according to any one of claims 61 to 98, wherein the conjugate has one of the following structures (IIc') or (IIc"): As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

101. The conjugate of any one of claims 61 to 99, wherein X 2 , X 3 Or both are CH or CF.

102. The conjugate of any one of claims 61 to 101, wherein X 1 , X 5 Or both are CR 3 , and R 3 is H or halogenated.

103. The conjugate of any one of claims 61 to 102, wherein X 1 , X 5 Or both are CR 3 , and R 3 It is halogenated.

104. The conjugate of any one of claims 61 to 102, wherein X 1 is CH, and X 5 It is CH.

105. The conjugate of any one of claims 61 to 102, wherein X 1 is CF, and X 5 It is CH.

106. The conjugate of any one of claims 61 to 102, wherein X 1 is CH, and X 5 It's CF.

107. The conjugate of any one of claims 61 to 98, wherein the conjugate has one of the following structures: (IIa-1), (IIa-2), (IIa-3), (IIa-4), (IIa-5), (IIa-6), (IIa-7) or (IIa-8): in: L b is independently at each occurrence a direct bond, an optionally substituted alkylene linker, an optionally substituted heteroalkylene linker, a heteroatom linker, or a combination thereof; and q6 is 0, 1 or 2, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or A pharmaceutically acceptable salt, solvate or prodrug thereof.

108. The conjugate of any one of claims 61 to 98, wherein the conjugate has one of the following structures: (IIc-1), (IIc-2), (IIc-3) or (IIc-4): in: L b is independently at each occurrence a direct bond, an optionally substituted alkylene linker, an optionally substituted heteroalkylene linker, a heteroatom linker, or a combination thereof; and q7 is 1, 2 or 3, as a stereoisomer, enantiomer or tautomer thereof or a mixture thereof; or A pharmaceutically acceptable salt, solvate or prodrug thereof.

109. The conjugate of any one of claims 61 to 98, wherein the conjugate has the following structure: (IId), (IIe), (IIf) or (IIg): in: L b is independently at each occurrence a direct bond, an optionally substituted alkylene linker, an optionally substituted heteroalkylene linker, a heteroatom linker, or a combination thereof; q8 is 0, 1, or 2; and q9 is 0, 1 or 2, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or A pharmaceutically acceptable salt, solvate or prodrug thereof.

110. The conjugate of any one of claims 61 to 98, wherein the conjugate has one of the following structures (IIh) or (IIi): in: L b is independently at each occurrence a direct bond, an optionally substituted alkylene linker, an optionally substituted heteroalkylene linker, a heteroatom linker, or a combination thereof, as a stereoisomer, enantiomer or tautomer, or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

111. The conjugate of any one of claims 92 to 110, wherein L b is a direct bond, an optionally substituted alkylene linker or an optionally substituted heteroalkylene linker, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

112. The conjugate of any one of claims 92 to 111, wherein L b is a direct key or has one of the following structures: in: R b is independently at each occurrence hydrogen, alkyl, hydroxyalkyl or alkoxyalkyl, as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

113. The conjugate of any one of claims 61 to 98, wherein the conjugate has one of the following structures: in: R 1b is a chemotherapeutic, cytotoxic, or myeloid cell agonist; R 2b Has one of the following structures: L 1g Has one of the following structures: As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

114. The conjugate of any one of claims 62 to 110, wherein the conjugate has one of the following structures: As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

115. The conjugate of claim 114, wherein R 1a Has one of the following structures: As a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

116. The conjugate of claim 114, wherein R 1a Has one of the following structures: in: R' is hydrogen or has one of the following structures: in: R a' Is H or C 1-6 alkyl; R b' It is C 1-6 Alkyl or C 1-6 Alkoxy; R c' It is H, C 1-6 Alkyl, -CH2OH or C 1-6 Alkoxy; R d' Is H or C 1-6 Alkyl; or R e ' is H or C 1-6 alkyl.

117. The conjugate of any one of claims 61 to 116, wherein L 1 or L 2 Has the following structure: in: ** Shown with X 1 , X 2 , X 3 , X 4 or X 5 a bond of , as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

118. The conjugate of any one of claims 61 to 91, wherein X 2 It is CL 1 -R 1 , and X 3 It is CL 2 -R 2 , as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

119. The conjugate of any one of claims 61 to 91, wherein X 3 It is CL 1 -R 1 , and X 2 It is CL 2 -R 2 , as a stereoisomer, enantiomer or tautomer or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

120. The conjugate of any one of claims 61 to 119, wherein L 4 Has the following structure:

121. The conjugate of any one of claims 61 to 119, wherein L 4 Has the following structure:

122. The conjugate of any one of claims 61 to 119, wherein L 4 Has the following structure:

123. A conjugate having a structure selected from Table 2 as a stereoisomer, enantiomer or tautomer thereof, or a mixture thereof; or a pharmaceutically acceptable salt, solvate or prodrug thereof.

124. The conjugate of any one of claims 61 to 123, wherein the targeting moiety is an anti-CD40 antibody, an antibody selected from the group consisting of an anti-LRRC15 antibody, an anti-CTSK antibody, an anti-ADAM12 antibody, an anti-ITGA11 antibody, an anti-FAP antibody, an anti-NOX4 antibody, an anti-SGCD antibody, an anti-SYNDIG1 antibody, an anti-CDH11 antibody, an anti-PLPP4 antibody, an anti-SLC24A2 antibody, an anti-PDGFRB antibody, an anti-THY1 antibody, an anti-ANTXR1 antibody, an anti-GAS1 antibody, an anti-CALHM5 antibody, an anti-SDC1 antibody, an anti-HER2 antibody, an anti-TROP2 antibody, an anti-MSLN antibody, an anti-nectin 4 antibody, an anti-ASGR1 antibody, and an anti-MUC16 antibody.

125. The conjugate of claim 124, wherein the antibody is a full-length antibody.

126. The conjugate of claim 124, wherein the antibody is an antigen binding fragment.

127. The conjugate of claim 121, wherein the antibody is a humanized antibody.

128. The conjugate of any one of claims 61 to 127, wherein g is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

129. A pharmaceutical composition comprising the conjugate according to any one of claims 61 to 128 and a pharmaceutically acceptable excipient.

130. A method for treating cancer, comprising administering an effective amount of the conjugate of any one of claims 61 to 128 or the pharmaceutical composition of claim 115 to a subject in need thereof.

131. A method for treating infection, inflammation, autoimmune disorder or a combination thereof, the method comprising administering to a subject in need thereof an effective amount of the conjugate of any one of claims 61 to 128 or the pharmaceutical composition of claim 129.

132. The method of any one of claims 130 to 131, wherein the effective amount of the conjugate is administered intravenously, subcutaneously, or intratumorally.

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