WO2026006688A2 - Degrader antibody conjugates and uses thereof - Google Patents

Abstract

The invention provides antibody conjugate compositions of Formula I comprising an antibody linked by conjugation to one or more target protein binder and VHL ligand (TPI-VHL) moieties. The invention also provides TPI-VHL derivative intermediate compositions comprising a reactive functional group. Such intermediate compositions are suitable substrates for formation of the antibody conjugates through a linker or linking moiety. The invention further provides methods of treating diseases and disorders such as cancer with the antibody conjugates.

Description

15077.006WO1 DEGRADER ANTIBODY CONJUGATES AND USES THEREOF CROSS REFERENCE TO RELATED APPLICATIONS 5 This non-provisional application claims the benefit of priority to U.S. Provisional Applications No.63/665,421, filed 28 June 2024, which is incorporated by reference in its entirety. FIELD OF THE INVENTION The disclosure relates generally to antibody conjugate compositions, intermediates for 10 their manufacture, and methods of their use. The compositions are useful for facilitating intracellular degradation of target proteins. BACKGROUND OF THE INVENTION The ubiquitin proteasome system can be manipulated to conduct targeted degradation of specific proteins. Promoting the targeted degradation of pathogenic proteins using small 15 molecule degraders is a new modality in the treatment of diseases, including redirecting the activity of E3 ligases such as cereblon (CRBN) or VHL. Proteolysis targeting chimera compounds called PROTAC (Sakamoto, K. M., et al. (2001) Proc. Natl. Acad. Sci. USA 98:8554–8559; Sun, X. et al. (2019) Signal Transduct. Target. Ther.4:64; Schapira, M., et al (2019) Nat. Rev. Drug Discov.18:949–963) and “molecular glue” compounds (Yang, Z., et al 20 (2021) Cell Research 31:1315–1318; Tan, X. et al. (2007) Nature 446:640–645; Han, T. et al. (2017) Science 356, eaal3755) are two modes of TPD. PROTAC comprise three parts, including a ligand for binding a target protein, another ligand for recruiting an E3 ligase, and a linker to help anchor the target protein to the E3 ubiquitin ligase to promote its ubiquitination and subsequent proteasomal degradation. Similar to PROTAC, molecular glues can also cause 25 ubiquitination and degradation of a target protein. In contrast to PROTAC, molecular glues are small molecular weight compounds that trigger a compact protein–protein interaction between a target protein and an E3 ubiquitin ligase. Molecular glues are typically smaller than PROTAC and may have better pharmacological properties, higher membrane permeability, better cellular uptake, and better penetration of the blood-brain barrier. Molecular glues promote the poly- 30 ubiquitination and proteasomal degradation of various disease-associated protein targets (Chamberlain, P., et al (2019) Drug Disc. Today: Tech.31:29-34; WO 2022/152821). The molecular glue molecules bind to both the E3 ligase and the target protein, thereby mediating an alteration of the ligase surface and enabling an interaction with the target protein. Examples 1 15077.006WO1 include the IMiD (immunomodulatory imide drug) class including thalidomide, lenalidomide and pomalidomide, each approved for use in treating hematological cancers. More efficient targeting strategies are still required. Receptor-interacting protein kinase 2 (RIPK2) is a serine/threonine kinase integral to 5 innate immune signaling via the NOD1/2 pathways, with emerging roles in tumor biology. RIPK2 activation promotes chronic inflammation through NF-κB and MAPK signaling, fostering tumor progression by enhancing immune cell infiltration, RNA methylation, and tumor stemness (Yan et al., (2022) Nat. Comm.13:10384). In gastric cancer, RIPK2 overexpression correlates with immune checkpoint biomarkers (e.g., PD-L1), microsatellite instability, and 10 tumor mutational burden, suggesting its role in immunotherapy resistance (Zhang et al., (2024) Jour. of Cancer 15:176–191). Preclinical studies demonstrate that RIPK2 inhibition suppresses metastasis by destabilizing YAP via ITCH-mediated ubiquitination, highlighting its potential as a therapeutic target (Yan et al., (2025) Cell Death & Disease 16:27594). Its dual role in immune modulation and tumor cell survival underscores its relevance in oncology, particularly for 15 cancers with inflammatory microenvironments. The Bcl-2 family includes more than 20 members of pro-survival and pro-apoptotic proteins. Bcl-2, Bcl-xL, Mcl-1, and Bcl-W are pro-survival proteins which promote cell survival by inhibiting their pro-apoptotic counterparts. Members in the pro-survival subgroup have four BH (Bcl-2 Homology) domains (BH1～BH4), except that Mcl-1 only has BH 1, 2, 3 domains 20 (Li M. et al (2020) Pharm. Res.151:104547). Another subgroup of Bcl-2 family is pro-apoptotic proteins and can be further classified as BH domain proteins (Bax, Bak, and Bok) , which contain three BH domains (BH1～BH3) and function as effectors of apoptosis; the other is BH3- only proteins (Bad, Bid, Bim, Noxa, PUMA, Bmf, Hrk, and Bik) which are the initiators of apoptosis. Bcl-2 family members work cooperatively to govern cell fate, as in healthy condition, 25 the pro-survival members bind to Bax and Bak to restrict the oligomerization of Bax/Bak, impairing their ability to induce apoptotic pores formation and permeabilization of the outer mitochondrial membrane. BH3-only proteins, as induced transcriptionally or post- transcriptionally by apoptotic stimuli, promote apoptosis by binding competitively to pro- survival Bcl-2 family members to release Bax/Bak or by directly activating these effector 30 proteins (Hata, Engelman et al.2015) . Dysregulation of apoptosis, usually caused by the imbalance between pro-survival and pro-apoptotic proteins in the Bcl-2 family, leads to uncontrolled cell growth and tumor development. Pro-survival protein Bcl-xL (B-cell lymphoma-extra large), one of the key regulators of the intrinsic pathway, (Boise, Gonzalez-Garcia et al. (1993) Cell 74(4):597-608), shares 44% 35 homology in amino acid sequence with Bcl-2 and has a similar structural domain to Bcl-2. A 2 15077.006WO1 hydrophobic pocket formed by the BH1–BH3 domains of Bcl-xL interacts with the BH3 domain of the pro-apoptotic proteins to form a heterodimer. In addition, the BH4 domain of Bcl-xL is involved in its anti-apoptotic activity (Lewis, Hayashi et al.2014, Lee and Fairlie 2019). Since Bcl-xL is the most common Bcl-2 family member overexpressed in solid tumors, as well as in 5 some subsets of leukemia and lymphoma, it is highly desirable to develop a therapeutic strategy that can retain the benefit of targeting Bcl-xL. Given the importance of Bcl-xL in regulating apoptosis, there remains a need for therapies that inhibit Bcl-xL activity, particularly selectively, as an approach towards the treatment of diseases in which apoptosis is dysregulated via expression or over-expression of anti-apoptotic Bcl-2 family proteins, such as Bcl-xL. Targeted 10 therapeutic agents to treat hyperproliferative disorders like cancer, and other disease are of interest. KRAS (Kirsten rat sarcoma virus) is a proto-oncogene that encodes the K‑Ras protein, a small GTPase that is a critical component of the RAS/MAPK signaling cascade. This protein normally functions as a molecular switch cycling between an inactive GDP‑bound state and an 15 active GTP‑bound state. When activated, KRas transmits signals from cell surface receptors (e.g. through cRaf and phosphatidylinositol‑3‑kinase [PI3K]) to the nucleus, thereby regulating cell proliferation, differentiation, and survival. KRas also influences metabolic reprogramming in cancer cells (for example, by upregulating the GLUT1 glucose transporter and contributing to the Warburg effect). Mutations, typically single nucleotide substitutions at key codons such as 20 G12, G13, and Q61, lock KRas in its active state and drive the development of a wide range of malignancies, including lung adenocarcinoma, colorectal cancer, and pancreatic ductal adenocarcinoma. It is important to note that the KRAS gene produces two splice variants, KRAS4A and KRAS4B, that differ mainly in their C‑terminal regions and membrane interactions. KRAS4b is typically expressed at much higher levels in most human tissues and is 25 the predominant form in many cancers. Therapeutic strategies and drug development efforts have largely focused on KRAS4B. The prevalence and central role of KRAS mutations in oncogenesis have made it an attractive drug target because of its high affinity for GTP/GDP although the absence of deep, well-defined binding pockets has made it difficult to target with traditional small molecules. 30 WEE1, a tyrosine kinase regulating the G2/M checkpoint, ensures genomic stability by phosphorylating CDK1/2. Overexpression in endometrial cancer and melanoma confers poor prognosis by enabling DNA repair evasion and fostering replication stress (Magnussen et al., (2012) PLOS ONE 7:e38254). WEE1 inhibitors (e.g., adavosertib) synergize with DNA- damaging agents to induce mitotic catastrophe, particularly in TP53-mutant tumors (Liao et al.,35 (2024) Jour. of Cancer 15:545–559). Recent studies reveal WEE1’s role in activating STING- 3 15077.006WO1 dependent innate immune responses, enhancing anti-tumor immunity via IFN-β secretion (Nielsen et al., (2023) Clinical Cancer Research 29:4321–4333). Its dual function in cell cycle control and immune modulation makes WEE1 a promising target for precision oncology, especially in replication-stress-prone cancers. 5 The Von Hippel-Lindau (VHL) protein is among the most widely recruited E3 ligases for protein degradation. Many potent small-molecule VHL binders feature a (R)- hydroxyproline motif (Buckley, D. L.; et al. J. Am. Chem. Soc. (2012) 134 (10):4465−4468; Galdeano, C.; et al J. Med. Chem. (2014) 57 (20), 8657−8663; Testa, A.; et al J. Am. Chem. Soc. (2018) 140 (29), 9299−9313; Han, X.; et al J. Med. Chem. (2019) 62 (24), 11218−11231) which 10 forms an interaction with Ser110 in the HIF1α binding site of VHL but limits passive transport across the cell membrane (Klein, V. G.; et al ACS Med. Chem. Lett. (2020) 11 (9), 1732−1738; Han, X.; et al (2022) Cell Rep. Phys. Sci.3 (10):101062; Shah, R. R.; et al (2020) Bioorg. Med. Chem.28 (5), 115326; Diehl, C. J. et al Chem. Soc. Rev. (2022) 51 (19):8216). Protein degraders which ligand the E3 ligase in a covalent manner offer potential advantages over their reversible 15 counterparts by transforming the ternary complex into a simple binary interaction between modified E3 and the substrate Targeted therapeutic agents to treat hyperproliferative disorders like cancer, and other disease are of interest. SUMMARY OF THE INVENTION 20 The invention is generally directed to an antibody conjugate composition comprising a target protein binder and VHL ligand moiety (TPI-VHL) covalently attached to an antibody by an antibody linker, wherein the antibody binds to a tumor-associated antigen or cell-surface receptor. The antibody conjugate composition of claim 1 has Formula I: 25 Ab−[L−(TPI−Sp−VHL)]_p I or a pharmaceutically acceptable salt thereof, wherein: Ab is the antibody; L is the antibody linker; 30 TPI-VHL is the target protein binder and VHL ligand moiety wherein the target protein binder is covalently attached to the VHL ligand by a spacer unit Sp; and p is an integer from 1 to 12. Another aspect of the invention is the VHL ligand has Formula Ia: 4 15077.006WO1 wherein the substituents are defined herein. Another aspect of the invention is the antibody conjugate composition prepared by conjugation of a cysteine amino acid of an antibody with a TPI-VHL linker compound (TVL). 5 Another aspect of the invention is a process for preparing the antibody conjugate comprising reacting a cysteine amino acid of an antibody with a TPI-VHL linker compound. Another aspect of the invention is a pharmaceutical composition comprising a therapeutically effective amount of the antibody conjugate composition, and one or more pharmaceutically acceptable diluent, vehicle, carrier or excipient. 10 Another aspect of the invention is a method for treating cancer comprising administering a therapeutically effective amount of the pharmaceutical composition to a patient in need thereof, Another aspect of the invention is a use of the antibody conjugate composition in the manufacture of a medicament for the treatment of cancer in a mammal. 15 Another aspect of the invention is the antibody conjugate composition for use in a method for treating cancer. DETAILED DESCRIPTION OF THE INVENTION Reference will now be made in detail to certain embodiments of the invention, examples of which are illustrated in the accompanying structures and formulas. While the invention will 20 be described in conjunction with the enumerated embodiments, it will be understood that they are not intended to limit the invention to those embodiments. On the contrary, the invention is intended to cover all alternatives, modifications, and equivalents, which may be included within the scope of the invention as defined by the claims. One skilled in the art will recognize many methods and materials similar or equivalent to 25 those described herein, which could be used in the practice of the present invention. The invention is in no way limited to the methods and materials described. 5 15077.006WO1 DEFINITIONS The terms “antibody” or “antibody construct” refer to a polypeptide comprising an antigen binding region (including the complementarity determining region (CDRs)) from an immunoglobulin gene or fragments thereof. The term “antibody” specifically encompasses 5 monoclonal antibodies (including full length monoclonal antibodies), polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments that exhibit the desired biological activity. An exemplary immunoglobulin (antibody) structural unit comprises a tetramer. Each tetramer is composed of two identical pairs of polypeptide chains, each pair having one “light” (about 25 kDa) and one “heavy” chain (about 50-70 kDa) connected by 10 disulfide bonds. Each chain is composed of structural domains, which are referred to as immunoglobulin domains. These domains are classified into different categories by size and function, e.g., variable domains or regions on the light and heavy chains (V_L and V_H, respectively) and constant domains or regions on the light and heavy chains (C_L and C_H, respectively). The N-terminus of each chain defines a variable region of about 100 to 110 or 15 more amino acids, referred to as the paratope, primarily responsible for antigen recognition, i.e., the antigen binding domain. Light chains are classified as either kappa or lambda. Heavy chains are classified as gamma, mu, alpha, delta, or epsilon, which in turn define the immunoglobulin classes, IgG, IgM, IgA, IgD and IgE, respectively. IgG antibodies are large molecules of about 150 kDa composed of four peptide chains. IgG antibodies contain two identical class γ heavy 20 chains of about 50 kDa and two identical light chains of about 25 kDa, thus a tetrameric quaternary structure. The two heavy chains are linked to each other and to a light chain each by disulfide bonds. The resulting tetramer has two identical halves, which together form the Y-like shape. Each end of the fork contains an identical antigen binding domain. There are four IgG subclasses (IgG1, IgG2, IgG3, and IgG4) in humans, named in order of their abundance in 25 serum (i.e., IgG1 is the most abundant). Typically, the antigen binding domain of an antibody will be most critical in specificity and affinity of binding to cancer cells. “Bispecific” antibodies (bsAbs) are antibodies that bind two distinct epitopes to cancer (Suurs F.V. et al (2019) Pharmacology & Therapeutics 201:103-119). Bispecific antibodies may engage immune cells to destroy tumor cells, deliver TPI-VHL moieties to tumors, and/or block 30 tumor signaling pathways. An antibody that targets a particular antigen includes a bispecific or multispecific antibody with at least one antigen binding region that targets the particular antigen. In some embodiments, the targeted monoclonal antibody is a bispecific antibody with at least one antigen binding region that targets tumor cells. Such antigens include but are not limited to: mesothelin, prostate specific membrane antigen (PSMA), HER2, TROP2, CEA, CEACAM5, 35 EGFR, 5T4, Nectin4, CCL-1, CCR7, CD19, CD20, CD22, CD30, CD33, CD70, CD79b, 6 15077.006WO1 CD123, CDH3, B7H3, B7H4 (also known as 08E), Integrin-beta6, protein tyrosine kinase 7 (PTK7), glypican-3, GPC-1, LIV-1, Folate receptor alpha, Claudin18.2, RG1, fucosyl-GMl, tissue factor (CD142), cKit (CD117), Axl, , GC-C, CTLA-4, and CD44 (WO 2017/196598). In some embodiments, the antibody construct is an antigen-binding antibody “fragment,” 5 which comprises at least an antigen-binding region of an antibody, alone or with other components that together constitute the antibody construct. Many different types of antibody “fragments” are known in the art, including, for instance, (i) a Fab fragment, which is a monovalent fragment consisting of the VL, VH, CL, and CH1 domains, (ii) a F(ab’)2 fragment, which is a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the 10 hinge region, (iii) a Fv fragment consisting of the V_L and V_H domains of a single arm of an antibody, (iv) a Fab’ fragment, which results from breaking the disulfide bridge of an F(ab’)2 fragment using mild reducing conditions, (v) a disulfide-stabilized Fv fragment (dsFv), and (vi) a single chain Fv (scFv), which is a monovalent molecule consisting of the two domains of the Fv fragment (i.e., VL and VH) joined by a synthetic linker which enables the two domains to be 15 synthesized as a single polypeptide chain. In some embodiments, the antibody construct is an antibody or a fusion protein comprising (i) an antigen binding domain and (ii) an Fc domain. The antibody or antibody fragment can be part of a larger construct, for example, a conjugate or fusion construct of the antibody fragment to additional regions. For instance, in some embodiments, the antibody fragment can be fused to an Fc region as described herein. In 20 other embodiments, the antibody fragment (e.g., a Fab or scFv) can be part of a chimeric antigen receptor or chimeric T-cell receptor, for instance, by fusing to a transmembrane domain (optionally with an intervening linker or “stalk” (e.g., hinge region)) and optional intercellular signaling domain. For instance, the antibody fragment can be fused to the gamma and/or delta chains of a t-cell receptor, so as to provide a T-cell receptor like construct that binds PD-L1. In 25 yet another embodiment, the antibody fragment is part of a bispecific T-cell engager (BiTEs) comprising a CD1 or CD3 binding domain and linker. In some embodiments, the antibody construct comprises an Fc domain. In certain embodiments, the antibody construct is a fusion protein. The antigen binding domain can be a single-chain variable region fragment (scFv). A single-chain variable region fragment (scFv), 30 which is a truncated Fab fragment including the variable (V) domain of an antibody heavy chain linked to a V domain of a light antibody chain via a synthetic peptide, can be generated using routine recombinant DNA technology techniques. Similarly, disulfide-stabilized variable region fragments (dsFv) can be prepared by recombinant DNA technology. The antibody construct or antigen binding domain may comprise one or more variable regions (e.g., two variable regions) 7 15077.006WO1 of an antigen binding domain of an anti-CEA antibody, each variable region comprising a CDR1, a CDR2, and a CDR3. “Cysteine-mutant antibody” is an antibody in which one or more amino acid residues of an antibody are substituted with cysteine residues. A cysteine-mutant antibody may be prepared 5 from the parent antibody by antibody engineering methods (Junutula, J. et al., (2008b) Nature Biotech., 26(8):925-932; Dornan et al. (2009) Blood 114(13):2721-2729; US 7521541; US 7723485; US 2012/0121615; WO 2009/052249). Cysteine residues provide for site-specific conjugation of a TPI-VHL compound to the antibody through the reactive cysteine thiol groups at the engineered cysteine sites but do not perturb immunoglobulin folding and assembly or alter10 antigen binding and effector functions. Cysteine-mutant antibodies can be conjugated to the TPI- VHL-linker compound with uniform stoichiometry of the antibody conjugate (e.g., up to two TPI-VHL moieties per antibody in an antibody that has a single engineered, mutant cysteine site). The TPI-VHL-linker compound has a reactive electrophilic group to react specifically with the free cysteine thiol groups of the cysteine-mutant antibody. 15 “Epitope” means any antigenic determinant or epitopic determinant of an antigen to which an antigen binding domain binds (i.e., at the paratope of the antigen binding domain). Antigenic determinants usually consist of chemically active surface groupings of molecules, such as amino acids or sugar side chains, and usually have specific three dimensional structural characteristics, as well as specific charge characteristics. 20 The terms “Fc receptor” or “FcR” refer to a receptor that binds to the Fc region of an antibody. There are three main classes of Fc receptors: (1) FcγR which bind to IgG, (2) FcαR which binds to IgA, and (3) FcεR which binds to IgE. The FcγR family includes several members, such as FcγI (CD64), FcγRIIA (CD32A), FcγRIIB (CD32B), FcγRIIIA (CD16A), and FcγRIIIB (CD16B). The Fcγ receptors differ in their affinity for IgG and also have different 25 affinities for the IgG subclasses (e.g., IgG1, IgG2, IgG3, and IgG4). Nucleic acid or amino acid sequence “identity,” as referenced herein, can be determined by comparing a nucleic acid or amino acid sequence of interest to a reference nucleic acid or amino acid sequence. The percent identity is the number of nucleotides or amino acid residues that are the same (i.e., that are identical) as between the optimally aligned sequence of interest 30 and the reference sequence divided by the length of the longest sequence (i.e., the length of either the sequence of interest or the reference sequence, whichever is longer). Alignment of sequences and calculation of percent identity can be performed using available software programs. Examples of such programs include CLUSTAL-W, T-Coffee, and ALIGN (for alignment of nucleic acid and amino acid sequences), BLAST programs (e.g., BLAST 2.1, 35 BL2SEQ, BLASTp, BLASTn, and the like) and FASTA programs (e.g., FASTA3x, FASTM, and 8 15077.006WO1 SSEARCH) (for sequence alignment and sequence similarity searches). Sequence alignment algorithms also are disclosed in, for example, Altschul et al., J. Molecular Biol., 215(3): 403-410 (1990), Beigert et al., Proc. Natl. Acad. Sci. USA, 106(10): 3770-3775 (2009), Durbin et al., eds., Biological Sequence Analysis: Probalistic Models of Proteins and Nucleic Acids, 5 Cambridge University Press, Cambridge, UK (2009), Soding, Bioinformatics, 21(7): 951-960 (2005), Altschul et al., Nucleic Acids Res., 25(17): 3389-3402 (1997), and Gusfield, Algorithms on Strings, Trees and Sequences, Cambridge University Press, Cambridge UK (1997)). Percent (%) identity of sequences can be also calculated, for example, as 100 x [(identical positions)/min(TG_A, TG_B)], where TG_A and TG_B are the sum of the number of residues and 10 internal gap positions in peptide sequences A and B in the alignment that minimizes TG_A and TGB. See, e.g., Russell et al., J. Mol Biol., 244: 332-350 (1994). The “antibody construct” or “binding agent” comprises Ig heavy and light chain variable region polypeptides that together form the antigen binding site. Each of the heavy and light chain variable regions are polypeptides comprising three complementarity determining regions 15 (CDR1, CDR2, and CDR3) connected by framework regions. The antibody construct can be any of a variety of types of binding agents known in the art that comprise Ig heavy and light chains. For instance, the binding agent can be an antibody, an antigen-binding antibody “fragment,” or a T-cell receptor. “Amino acid” refers to any monomeric unit that can be incorporated into a peptide, 20 polypeptide, or protein. Amino acids include naturally-occurring α-amino acids and their stereoisomers, as well as unnatural (non-naturally occurring) amino acids and their stereoisomers. “Stereoisomers” of a given amino acid refer to isomers having the same molecular formula and intramolecular bonds but different three-dimensional arrangements of bonds and atoms (e.g., an L-amino acid and the corresponding D-amino acid). The amino acids 25 can be glycosylated (e.g., N-linked glycans, O-linked glycans, phosphoglycans, C-linked glycans, or glypication) or deglycosylated. Amino acids may be referred to herein by either the commonly known three letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Naturally-occurring amino acids are those encoded by the genetic code, as well as those 30 amino acids that are later modified, e.g., hydroxyproline, γ-carboxyglutamate, and O-phosphoserine. Naturally-occurring α-amino acids include, without limitation, alanine (Ala), cysteine (Cys), aspartic acid (Asp), glutamic acid (Glu), phenylalanine (Phe), glycine (Gly), histidine (His), isoleucine (Ile), arginine (Arg), lysine (Lys), leucine (Leu), methionine (Met), asparagine (Asn), proline (Pro), glutamine (Gln), serine (Ser), threonine (Thr), valine (Val), 35 tryptophan (Trp), tyrosine (Tyr), and combinations thereof. Stereoisomers of naturally-occurring 9 15077.006WO1 α-amino acids include, without limitation, D-alanine (D-Ala), D-cysteine (D-Cys), D-aspartic acid (D-Asp), D-glutamic acid (D-Glu), D-phenylalanine (D-Phe), D-histidine (D-His), D- isoleucine (D-Ile), D-arginine (D-Arg), D-lysine (D-Lys), D-leucine (D-Leu), D-methionine (D- Met), D-asparagine (D-Asn), D-proline (D-Pro), D-glutamine (D-Gln), D-serine (D-Ser), D- 5 threonine (D-Thr), D-valine (D-Val), D-tryptophan (D-Trp), D-tyrosine (D-Tyr), and combinations thereof. Naturally-occurring amino acids include those formed in proteins by post-translational modification, such as citrulline (Cit). Unnatural (non-naturally occurring) amino acids include, without limitation, amino acid 10 analogs, amino acid mimetics, synthetic amino acids, N-substituted glycines, and N-methyl amino acids in either the L- or D-configuration that function in a manner similar to the naturally- occurring amino acids. For example, “amino acid analogs” can be unnatural amino acids that have the same basic chemical structure as naturally-occurring amino acids (i.e., a carbon that is bonded to a hydrogen, a carboxyl group, an amino group) but have modified side-chain groups 15 or modified peptide backbones, e.g., homoserine, norleucine, methionine sulfoxide, and methionine methyl sulfonium. “Amino acid mimetics” refer to chemical compounds that have a structure that is different from the general chemical structure of an amino acid, but that functions in a manner similar to a naturally-occurring amino acid. “Linker” refers to a functional group that covalently bonds two or more moieties in an 20 antibody conjugate compound. For example, the linking moiety can serve to covalently bond a drug TPI-VHL moiety to an antibody in an antibody conjugate composition. Useful bonds for connecting linking moieties to proteins and other materials include, but are not limited to, amides, amines, esters, carbamates, ureas, thioethers, thiocarbamates, thiocarbonates, and thioureas. 25 “Divalent” refers to a chemical moiety that contains two points of attachment for linking two functional groups; polyvalent linking moieties can have additional points of attachment for linking further functional groups. Divalent radicals may be denoted by the suffix “diyl”. For example, divalent linking moieties include divalent polymer moieties such as divalent poly(ethylene glycol), divalent cycloalkyl, divalent heterocycloalkyl, divalent aryl, and divalent 30 heteroaryl group. A “divalent cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group” refers to a cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group having two points of attachment for covalently linking two moieties in a molecule or material. Cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups can be substituted or unsubstituted. Cycloalkyl, heterocycloalkyl, aryl, or heteroaryl groups can be substituted with one or more groups selected from halo, hydroxy, 35 amino, alkylamino, amido, acyl, nitro, cyano, alkoxy, and others. 10 15077.006WO1 A _{wavy line (“ ”) and one or more asterisks (*) represents a point of attachment of} the specified chemical moiety to another moiety. If the specified chemical moiety has two wavy l_{ines (“ ”) present, it will be understood that the chemical moiety can be used bilaterally, i.e.,} as read from left to right or from right to left. In some embodiments, a specified moiety having 5 _{two wavy lines (“ ”) present is considered to be used as read from left to right.} “Alkyl” refers to a straight (linear) or branched, saturated, aliphatic radical having the number of carbon atoms indicated. Alkyl can include any number of carbons, for example from one to six, one to eight, one to twelve, one to twenty, or one to forty. Examples of alkyl groups include, but are not limited to, methyl (Me, -CH3), ethyl (Et, -CH2CH3), 1-propyl (n-Pr, n-10 propyl, -CH₂CH₂CH₃), 2-propyl (i-Pr, i-propyl, -CH(CH₃)₂), 1-butyl (n-Bu, n-butyl, - CH2CH2CH2CH3), 2-methyl-1-propyl (i-Bu, i-butyl, -CH2CH(CH3)2), 2-butyl (s-Bu, s-butyl, - CH(CH3)CH2CH3), 2-methyl-2-propyl (t-Bu, t-butyl, -C(CH3)3), 1-pentyl (n-pentyl, - CH₂CH₂CH₂CH₂CH₃), 2-pentyl (-CH(CH₃)CH₂CH₂CH₃), 3-pentyl (-CH(CH₂CH₃)₂), 2-methyl- 2-butyl (-C(CH₃)₂CH₂CH₃), 3-methyl-2-butyl (-CH(CH₃)CH(CH₃)₂), 3-methyl-1-butyl (-15 CH2CH2CH(CH3)2), 2-methyl-1-butyl (-CH2CH(CH3)CH2CH3), 1-hexyl (- CH₂CH₂CH₂CH₂CH₂CH₃), 2-hexyl (-CH(CH₃)CH₂CH₂CH₂CH₃), 3-hexyl (- CH(CH₂CH₃)(CH₂CH₂CH₃)), 2-methyl-2-pentyl (-C(CH₃)₂CH₂CH₂CH₃), 3-methyl-2-pentyl (- CH(CH3)CH(CH3)CH2CH3), 4-methyl-2-pentyl (-CH(CH3)CH2CH(CH3)2), 3-methyl-3-pentyl (- C(CH3)(CH2CH3)2), 2-methyl-3-pentyl (-CH(CH2CH3)CH(CH3)2), 2,3-dimethyl-2-butyl (- 20 C(CH₃)₂CH(CH₃)₂), 3,3-dimethyl-2-butyl (-CH(CH₃)C(CH₃)₃, 1-heptyl, 1-octyl, and the like. Alkyl groups can be substituted or unsubstituted. “Substituted alkyl” groups can be substituted with one or more groups such as halo, hydroxy, amino, oxo (=O), alkylamino, amido, acyl, nitro, cyano, and alkoxy. The term “alkyldiyl” refers to a divalent alkyl radical. Examples of alkyldiyl groups25 include, but are not limited to, methylene (-CH2-), ethylene (-CH2CH2-), propylene (- CH₂CH₂CH₂-), and the like. An alkyldiyl group may also be referred to as an “alkylene” group. “Alkenyl” refers to a straight (linear) or branched, unsaturated, aliphatic radical having the number of carbon atoms indicated and at least one carbon-carbon double bond, sp2. Alkenyl can include from two to about 12 or more carbons atoms. Alkenyl groups are radicals having 30 “cis” and “trans” orientations, or alternatively, “E” and “Z” orientations. Examples include, but are not limited to, ethylenyl or vinyl (-CH=CH2), allyl (-CH2CH=CH2). butenyl, pentenyl, and isomers thereof. Alkenyl groups can be substituted or unsubstituted. “Substituted alkenyl” groups can be substituted with one or more groups selected from halo, hydroxy, amino, oxo (=O), alkylamino, amido, acyl, nitro, cyano, and alkoxy. 11 15077.006WO1 The terms “alkenylene” or “alkenyldiyl” refer to a linear or branched-chain divalent hydrocarbon radical. Examples include, but are not limited to, ethylenylene or vinylene (- CH=CH-), allyl (-CH₂CH=CH-), and the like. “Alkynyl” refers to a straight (linear) or branched, unsaturated, aliphatic radical having 5 the number of carbon atoms indicated and at least one carbon-carbon triple bond, sp. Alkynyl can include from two to about 12 or more carbons atoms. For example, C2-C6 alkynyl includes, but is not limited to ethynyl (-C^CH), propynyl (propargyl, -CH₂C^CH), butynyl, pentynyl, hexynyl, and isomers thereof Alkynyl groups can be substituted or unsubstituted. “Substituted alkynyl” groups can be substituted with one or more groups selected from halo, hydroxy, amino, 10 oxo (=O), alkylamino, amido, acyl, nitro, cyano, and alkoxy. The term “alkynylene” or “alkynyldiyl” refer to a divalent alkynyl radical. "Heteroalkyl" or “heteroalkylene” refer to a monovalent, straight or branched chain alkyl group, as defined above, comprising at least one heteroatom including but not limited to Si, N, O, P or S within the alkyl chain or at a terminus of the alkyl chain. In 15 some embodiments, a heteroatom is within the alkyl chain. In other embodiments, a heteroatom is at a terminus of the alkylene and thus serves to join the alkyl to the remainder of the molecule. In some embodiments, a heteroalkyl group may have 1 to 12 carbon atoms (C₁-C₁₂ heteroalkyl). In some embodiments, a heteroalkyl group may have 1 to 24 carbon atoms (C1-C24 heteroalkyl). In some embodiments, a heteroalkyl group may 20 have 1 to 40 carbon atoms (C₁-C₄₀ heteroalkyl) or 1-60 carbon atoms (C₁-C₆₀ heteroalkyl). Unless stated otherwise specifically in the specification, a heteroalkyl group is optionally substituted. For example, heteroalkyl groups can be substituted with 1-6 fluoro (F) substituents, for example, on the carbon backbone (as −CHF− or −CF2−) or on terminal carbons of straight chain or branched heteroalkyls (such as −CHF₂ or −CF₃). Examples of25 heteroalkyl groups include, but are not limited to, −CH₂CH₂OCH₃, −CH₂CH₂NHCH₃, − CH₂CH₂N(CH₃)₂, −C(=O)NHCH₂CH₂NHCH₃, −C(=O)N(CH₃)CH₂CH₂N(CH₃)₂, − C(=O)NHCH₂CH₂NHC(=O)CH₂CH₃, −C(=O)N(CH₃)CH₂CH₂N(CH₃)C(=O)CH₂CH₃, − OCH₂CH₂CH₂NH(CH₃), −OCH₂CH₂CH₂N(CH₃)₂, −OCH₂CH₂CH₂NHC(=O)CH₂CH₃, − OCH₂CH₂CH₂N(CH₃)C(=O)CH₂CH₃, −CH₂CH₂CH₂NH(CH₃), −OCH₂CH₂CH₂N(CH₃)₂, 30 −CH₂CH₂CH₂NHC(=O)CH₂CH₃, −CH₂CH₂CH₂N(CH₃)C(=O)CH₂CH₃, −CH₂SCH₂CH₃, −CH₂CH₂S(O)CH₃, −NHCH₂CH₂NHC(=O)CH₂CH₃, −CH₂CH₂S(O)₂CH₃, − CH₂CH₂OCF₃, and −Si(CH₃)₃. Up to two heteroatoms may be consecutive, such as, for 12 15077.006WO1 example, −CH2NHOCH3 and −CH2OSi(CH3)3. A terminal polyethylene glycol (PEG) moiety is a type of heteroalkyl group. Exemplary heteroalkyl groups also include ethylene oxide (e.g., polyethylene oxide), propylene oxide, amino acid chains (i.e., short to medium length peptides such as containing 1-15 amino acids), and alkyl chains connected via a 5 variety of functional groups such as amides, disulfides, ketones, phosphonates, phosphates, sulfates, sulfones, sulfonamides, esters, ethers, -S-, carbamates, ureas, thioureas, anhydrides, or the like (including combinations thereof). In some embodiments, a heteroalkyl group includes a polyamino acid having 1-10 amino acids. In some embodiments, a heteroalkyl group includes a polyamino acid having 1-5 amino acids. 10 Heteroalkyl groups include a solubilizing unit comprising one or more groups of polyglycine, polysarcosine, polyethyleneoxy (PEG), and a glycoside, or combinations thereof. "Heteroalkenyl" refers to a heteroalkyl group, as defined above, that contains at least one carbon-carbon double bond. "Heteroalkynyl" refers to a heteroalkyl group, as 15 defined above, that contains at least one carbon-carbon triple bond. “Heteroalkyldiyl” refers to a divalent form of a heteroalkyl group as defined above. In some embodiments, a heteroalkyldiyl group may have 1 to 12 carbon atoms (C1- C₁₂ heteroalkyldiyl). In some embodiments, a heteroalkyldiyl group may have 1 to 24 carbon atoms (C1-C24 heteroalkyldiyl). In some embodiments, a heteroalkyldiyl group 20 may have 1 to 40 carbon atoms (C₁-C₄₀ heteroalkyldiyl) or 1-60 carbon atoms (C₁-C₆₀ heteroalkyldiyl). Examples of heteroalkyldiyl groups include, but are not limited to, − C_{H2CH2OCH2−, −CH2CH2OCF2−, −CH2CH2NHCH2−, −CH2OC(=O)NH−, − CH2OP(=O)(OH)OCH2−, −C(=O)NHCH2CH2NHCH2−, −} C(=O)N(CH3)CH2CH2N(CH3)CH2−, −C(=O)NHCH2CH2NHC(=O)CH2CH2−, − 25 C(=O)N(CH3)CH2CH2N(CH3)C(=O)CH2CH2−, −OCH2CH2OCH2CH2−, − OCH2CH2OCH2C(=O)−, −OCH2CH2OCH2CH2C(=O)−, −OCH2CH2NHCH2−, − OCH2CH2N(CH3)CH2−, −OCH2CH2CH2NHCH2−, −OCH2CH2CH2N(CH3)CH2−, − OCH2CH2CH2NHC(=O)CH2CH2−, −OCH2CH2CH2N(CH3)C(=O)CH2CH2−, − CH2CH2CH2NHCH2−-, −CH2CH2CH2N(CH3)CH2−, −CH2CH2CH2NHC(=O)CH2CH2−,30 −CH₂CH₂CH₂N(CH₃)C(=O)CH₂CH₂−, −CH₂CH₂NHC(=O)−, −CH₂CH₂N(CH₃)CH₂−, − CH₂CH₂N⁺(CH₃)₂−, −NHCH₂CH₂(NH₂)CH₂−, and −NHCH₂CH₂(NHCH₃)CH₂−. A 13 15077.006WO1 divalent polyethylene glycol (PEG) moiety with one to about 50 units of −OCH2CH2− is a type of heteroalkyldiyl group. “Heteroalkenyldiyl” refers to a divalent form of a heteroalkenyl group. “Heteroalkynyldiyl” refers to a divalent form of a heteroalkynyl group. 5 The terms “carbocycle”, “carbocyclyl”, “carbocyclic ring” and “cycloalkyl” refer to a saturated or partially unsaturated, monocyclic, fused bicyclic, bridged polycyclic ring, or linked by a bond to form a bicyclic assembly containing from 3 to 20 ring carbon atoms, or the number of carbon atoms indicated. Saturated monocyclic carbocyclic rings include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl. Saturated bicyclic 10 and polycyclic carbocyclic rings include, for example, norbornyl, [2.2.2] bicyclooctanyl, decahydronaphthalyl and adamantyl. Carbocyclic groups can also be partially unsaturated, having one or more double or triple bonds in the ring. Representative carbocyclic groups that are partially unsaturated include, but are not limited to, cyclobutenyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl (1,3- and 1,4-isomers), cycloheptenyl, cycloheptadienyl, cyclooctenyl, 15 cyclooctadienyl (1,3-, 1,4- and 1,5-isomers), norbornenyl, and norbornadienyl. The term “cycloalkyldiyl” refers to a divalent cycloalkyl radical. “Aryl” refers to a monovalent aromatic hydrocarbon radical of 6-20 carbon atoms (C6− C20) derived by the removal of one hydrogen atom from a single carbon atom of a parent aromatic ring system.. Aryl groups can be monocyclic, fused to form bicyclic or tricyclic 20 groups, or linked by a bond to form a biaryl group. Representative aryl groups include phenyl, naphthyl and biphenyl. Other aryl groups include benzyl, having a methylene linking group. Some aryl groups have from 6 to 12 ring members, such as phenyl, naphthyl or biphenyl. Other aryl groups have from 6 to 10 ring members, such as phenyl or naphthyl. The terms “arylene” or “aryldiyl” mean a divalent aromatic hydrocarbon radical of 6-20 25 carbon atoms (C6−C20) derived by the removal of two hydrogen atom from a two carbon atoms of a parent aromatic ring system. Some aryldiyl groups are represented in the exemplary structures as “Ar”. Aryldiyl includes bicyclic radicals comprising an aromatic ring fused to a saturated, partially unsaturated ring, or aromatic carbocyclic ring. Typical aryldiyl groups include, but are not limited to, radicals derived from benzene (phenyldiyl), substituted benzenes,30 naphthalene, anthracene, biphenylene, indenylene, indanylene, 1,2-dihydronaphthalene, 1,2,3,4- tetrahydronaphthyl, and the like. Aryldiyl groups are also referred to as “arylene”, and are optionally substituted with one or more substituents described herein. The terms “heterocycle,” “heterocyclyl” and “heterocyclic ring” are used interchangeably herein and refer to a saturated or a partially unsaturated (i.e., having one or 14 15077.006WO1 more double and/or triple bonds within the ring) carbocyclic radical of 3 to about 20 ring atoms in which at least one ring atom is a heteroatom selected from nitrogen, oxygen, phosphorus and sulfur, the remaining ring atoms being C, where one or more ring atoms is optionally substituted independently with one or more substituents described below. Heterocycles can be monocyclic, 5 fused to form bicyclic or tricyclic groups, or linked by a bond to form a biheterocyclic such as the 4-(piperidin-4-yl)piperazine group: A heterocycle may be a monocycle having 3 to 7 ring members (2 to 6 carbon atoms and 1 to 4 heteroatoms selected from N, O, P, and S) or a bicycle having 7 to 10 ring members (4 to 10 9 carbon atoms and 1 to 6 heteroatoms selected from N, O, P, and S), for example: a bicyclo [4,5], [5,5], [5,6], or [6,6] system. Heterocycles are described in Paquette, Leo A.; “Principles of Modern Heterocyclic Chemistry” (W.A. Benjamin, New York, 1968), particularly Chapters 1, 3, 4, 6, 7, and 9; “The Chemistry of Heterocyclic Compounds, A series of Monographs” (John Wiley & Sons, New York, 1950 to present), in particular Volumes 13, 14, 16, 19, and 28; and J. 15 Am. Chem. Soc. (1960) 82:5566. “Heterocyclyl” also includes radicals where heterocycle radicals are fused with a saturated, partially unsaturated ring, or aromatic carbocyclic or heterocyclic ring. Examples of heterocyclic rings include, but are not limited to, morpholin-4- yl, piperidin-1-yl, piperazinyl, piperazin-4-yl-2-one, piperazin-4-yl-3-one, pyrrolidin-1-yl, thiomorpholin-4-yl, S-dioxothiomorpholin-4-yl, azocan-1-yl, azetidin-1-yl, 20 octahydropyrido[1,2-a]pyrazin-2-yl, [1,4]diazepan-1-yl, pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino, thiomorpholino, thioxanyl, piperazinyl, homopiperazinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl, diazepinyl, thiazepinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, 25 pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl, pyrazolidinylimidazolinyl, imidazolidinyl, 3-azabicyco[3.1.0]hexanyl, 3- azabicyclo[4.1.0]heptanyl, azabicyclo[2.2.2]hexanyl, 3H-indolyl quinolizinyl and N-pyridyl ureas. Spiro heterocyclyl moieties are also included within the scope of this definition. Examples of spiro heterocyclyl moieties include azaspiro[2.5]octanyl and azaspiro[2.4]heptanyl. 30 Examples of a heterocyclic group wherein 2 ring atoms are substituted with oxo (=O) moieties are pyrimidinonyl and 1,1-dioxo-thiomorpholinyl. The heterocycle groups herein are optionally substituted independently with one or more substituents described herein. The term “heterocyclyldiyl” refers to a divalent, saturated or a partially unsaturated (i.e., having one or more double and/or triple bonds within the ring) carbocyclic radical of 3 to about 15 15077.006WO1 20 ring atoms in which at least one ring atom is a heteroatom selected from nitrogen, oxygen, phosphorus and sulfur, the remaining ring atoms being C, where one or more ring atoms is optionally substituted independently with one or more substituents as described. Examples of 5- membered and 6-membered heterocyclyldiyls include morpholinyldiyl, piperidinyldiyl, 5 piperazinyldiyl, pyrrolidinyldiyl, dioxanyldiyl, thiomorpholinyldiyl, and S- dioxothiomorpholinyldiyl. The term “heteroaryl” refers to a monovalent aromatic radical of 5-, 6-, or 7-membered rings, and includes fused ring systems (at least one of which is aromatic) of 5-20 atoms, containing one or more heteroatoms independently selected from nitrogen, oxygen, and sulfur. 10 Heteroaryls can be monocyclic, fused to form bicyclic or tricyclic groups, or linked by a bond to form a biheteroaryl such as the 4-(pyrazol-3-yl)pyridine group: Examples of heteroaryl groups are pyridinyl (including, for example, 2- hydroxypyridinyl), imidazolyl, imidazopyridinyl, pyrimidinyl (including, for example, 4- 15 hydroxypyrimidinyl), pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxadiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, tetrahydroisoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, thiadiazolyl, furazanyl, benzofurazanyl, benzothiophenyl, benzothiazolyl, 20 benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, imidazo[1,2-a]pyridinyl, and furopyridinyl. Heteroaryl groups are optionally substituted independently with one or more substituents described herein. The term “heteroaryldiyl” refers to a divalent aromatic radical of 5-, 6-, or 7-membered rings, and includes fused ring systems (at least one of which is aromatic) of 5-20 atoms, 25 containing one or more heteroatoms independently selected from nitrogen, oxygen, and sulfur. Examples of 5-membered and 6-membered heteroaryldiyls include pyridyldiyl, imidazolyldiyl, pyrimidinyldiyl, pyrazolyldiyl, triazolyldiyl, pyrazinyldiyl, tetrazolyldiyl, furyldiyl, thienyldiyl, isoxazolyldiyldiyl, thiazolyldiyl, oxadiazolyldiyl, oxazolyldiyl, isothiazolyldiyl, imidazo[1,2- a]pyridindiyl, and pyrrolyldiyl. 30 The heterocycle or heteroaryl groups may be carbon (carbon-linked), or nitrogen (nitrogen-linked) bonded where such is possible. By way of example and not limitation, carbon bonded heterocycles or heteroaryls are bonded at position 2, 3, 4, 5, or 6 of a pyridine, position 3, 4, 5, or 6 of a pyridazine, position 2, 4, 5, or 6 of a pyrimidine, position 2, 3, 5, or 6 of a 16 15077.006WO1 pyrazine, position 2, 3, 4, or 5 of a furan, tetrahydrofuran, thiofuran, thiophene, pyrrole or tetrahydropyrrole, position 2, 4, or 5 of an oxazole, imidazole or thiazole, position 3, 4, or 5 of an isoxazole, pyrazole, or isothiazole, position 2 or 3 of an aziridine, position 2, 3, or 4 of an azetidine, position 2, 3, 4, 5, 6, 7, or 8 of a quinoline or position 1, 3, 4, 5, 6, 7, or 8 of an 5 isoquinoline. By way of example and not limitation, nitrogen bonded heterocycles or heteroaryls are bonded at position 1 of an aziridine, azetidine, pyrrole, pyrrolidine, 2-pyrroline, 3-pyrroline, imidazole, imidazolidine, 2-imidazoline, 3-imidazoline, pyrazole, pyrazoline, 2-pyrazoline, 3- pyrazoline, piperidine, piperazine, indole, indoline, 1H-indazole, position 2 of a isoindole, or 10 isoindolinone, position 4 of a morpholine, and position 9 of a carbazole, or β-carboline. The terms “halo” and “halogen,” by themselves or as part of another substituent, refer to a fluorine, chlorine, bromine, or iodine atom. The term “carbonyl,” by itself or as part of another substituent, refers to C(=O) or – C(=O)–, i.e., a carbon atom double-bonded to oxygen and bound to two other groups in the 15 moiety having the carbonyl. As used herein, the phrase “quaternary ammonium salt” refers to a tertiary amine that has been quaternized with an alkyl substituent (e.g., a C1-C4 alkyl such as methyl, ethyl, propyl, or butyl). The term “fused” refers to a ring which is joint to an adjacent ring and share two adjacent 20 ring atoms that form a covalent bond. The term “bridged” refers to a ring fusion wherein non-adjacent atoms on a ring are joined by a divalent substituent, such as alkylenyl group, an alkylenyl group containing one or two heteroatoms, or a single heteroatom. Quinuclidinyl and admantanyl are examples of bridged ring systems. 25 The term or prefix “spiro” refers to a ring substituent which is joined by two bonds at the same carbon atom. Examples of spiro groups include 1,1 -diethylcyclopentane, dimethyl- dioxolane, and 4-benzyl-4- methylpiperidine, wherein the cyclopentane and piperidine, respectively, are the spiro substituents. The terms “optional” or “optionally” means that the subsequently described event or 30 circumstance may or may not occur, and that the description includes instances where said event or circumstance occurs and instances in which it does not. Also, the term “optionally substituted” refers to any one or more hydrogen atoms on the designated atom or group may or may not be replaced by a moiety other than hydrogen. “Optionally substituted” may be zero to the maximum number of possible substitutions, and each occurrence is independent. When the 35 term “substituted” is used, then that substitution is required to be made at a substitutable 17 15077.006WO1 hydrogen atom of the indicated substituent. An optional substitution may be the same or different from a (required) substitution. The term "chiral" refers to molecules which have the property of non-superimposability of the mirror image partner, while the term "achiral" refers to molecules which are 5 superimposable on their mirror image partner. The term "stereoisomers" refers to compounds which have identical chemical constitution, but differ with regard to the arrangement of the atoms or groups in space. Stereochemical definitions and conventions used herein generally follow S. P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New 10 York; and Eliel, E. and Wilen, S., "Stereochemistry of Organic Compounds", John Wiley & Sons, Inc., New York, 1994. The compounds of the invention may contain asymmetric or chiral centers, and therefore exist in different stereoisomeric forms. It is intended that all stereoisomeric forms of the compounds of the invention, including but not limited to, diastereomers, enantiomers and atropisomers, as well as mixtures thereof such as racemic 15 mixtures, form part of the present invention. Many organic compounds exist in optically active forms, i.e., they have the ability to rotate the plane of plane-polarized light. In describing an optically active compound, the prefixes D and L, or R and S, are used to denote the absolute configuration of the molecule about its chiral center(s). The prefixes d and l or (+) and (-) are employed to designate the sign of rotation of plane-polarized light by the compound, with (-) or 20 1 meaning that the compound is levorotatory. A compound prefixed with (+) or d is dextrorotatory. For a given chemical structure, these stereoisomers are identical except that they are mirror images of one another. A specific stereoisomer may also be referred to as an enantiomer, and a mixture of such isomers is often called an enantiomeric mixture. A 50:50 mixture of enantiomers is referred to as a racemic mixture or a racemate, which may occur 25 where there has been no stereoselection or stereospecificity in a chemical reaction or process. The terms "racemic mixture" and "racemate" refer to an equimolar mixture of two enantiomeric species, devoid of optical activity. "Diastereomer" refers to a stereoisomer with two or more centers of chirality and whose molecules are not mirror images of one another. Diastereomers have different physical 30 properties, e.g. melting points, boiling points, spectral properties, and reactivities. Mixtures of diastereomers may separate under high resolution analytical procedures such as electrophoresis and chromatography. "Enantiomers" refer to two stereoisomers of a compound which are non-superimposable mirror images of one another. 35 The term "tautomer" or "tautomeric form" refers to structural isomers of different 18 15077.006WO1 energies which are interconvertible via a low energy barrier. For example, proton tautomers (also known as prototropic tautomers) include interconversions via migration of a proton, such as keto-enol and imine-enamine isomerizations. Valence tautomers include interconversions by reorganization of some of the bonding electrons. 5 The term "salt" refers to acid or base salts of the compounds of the disclosed herein. Illustrative examples of pharmaceutically acceptable salts are mineral acid (hydrochloric acid, hydrobromic acid, phosphoric acid, and the like) salts, organic acid (acetic acid, propionic acid, glutamic acid, citric acid and the like) salts, quaternary ammonium (methyl iodide, ethyl iodide, and the like) salts. It is understood that the pharmaceutically acceptable salts are non-toxic. 10 Pharmaceutically acceptable salts of the acidic compounds disclosed herein are salts formed with bases, namely cationic salts such as alkali and alkaline earth metal salts, such as sodium, lithium, potassium, calcium, magnesium, as well as ammonium salts, such as ammonium, trimethyl-ammonium, diethylammonium, and tris-(hydroxymethyl)-methyl-ammonium salts. Similarly acid addition salts, such as of mineral acids, organic carboxylic and organic sulfonic 15 acids, e.g., hydrochloric acid, methanesulfonic acid, maleic acid, are also possible provided a basic group, such as pyridyl, constitutes part of the structure. The neutral forms of the compounds can be regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents, but 20 otherwise the salts are equivalent to the parent form of the compound for the purposes of the present disclosure. Any compound or Formula given herein, is intended to represent unlabeled forms as well as isotopically labeled forms of the compounds (i.e., "isotopic analogs"). Isotopically labeled compounds have structures depicted by the formulas given herein except that one or more atoms 25 are replaced by an atom having a selected atomic mass or mass number. Examples of isotopes that can be incorporated into the disclosed compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, chlorine and iodine, such as ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ¹³N, 1⁵N, ¹⁵O, ¹⁷O, ¹⁸O, ³¹P, ³²P, ³⁵S, ¹⁸F, ³⁶Cl, ¹²³I and ¹²⁵I, respectively. Various isotopically labeled compounds of the present disclosure, for example those into which radioactive isotopes such as 30 ³H, ¹³C and ¹⁴C are incorporated. Such isotopically labeled compounds may be useful for enhanced therapeutic activity, in metabolic studies, reaction kinetic studies, detection or imaging techniques, such as positron emission tomography (PET) or single-photon emission computed tomography (SPECT) including drug or substrate tissue distribution assays or in radioactive treatment of patients. 19 15077.006WO1 The disclosure also includes "deuterated analogs" of compounds described herein in which from 1 to n hydrogens attached to a carbon atom is/are replaced by deuterium (²H), in which n is the number of hydrogens in the molecule. Such compounds may exhibit increased resistance to metabolism and are thus useful for increasing the half-life of any compound when 5 administered to a mammal, particularly a human. See, for example, Foster, "Deuterium Isotope Effects in Studies of Drug Metabolism," Trends Pharmacol. Sci.5(12):524-527 (1984). Such compounds are synthesized by means well known in the art, for example by employing starting materials in which one or more hydrogens have been replaced by deuterium. Deuterium labeled or substituted therapeutic compounds of the disclosure may have improved DMPK (drug 10 metabolism and pharmacokinetics) properties, relating to distribution, metabolism and excretion (ADME). Substitution with heavier isotopes such as deuterium may afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life, reduced dosage requirements and/or an improvement in therapeutic index. An ¹⁸F, ³H, or ¹¹C labeled compound may be useful for PET or SPECT or other imaging studies. Isotopically 15 labeled compounds of this disclosure and prodrugs thereof can generally be prepared by carrying out the procedures disclosed in the schemes or in the examples and preparations described below by substituting a readily available isotopically labeled reagent for a non-isotopically labeled reagent. It is understood that deuterium in this context is regarded as a substituent in a compound described herein. The concentration of such a heavier isotope, specifically deuterium, 20 may be defined by an isotopic enrichment factor. In the compounds of this disclosure any atom not specifically designated as a particular isotope is meant to represent any stable isotope of that atom. Unless otherwise stated, when a position is designated specifically as "H" or "hydrogen", the position is understood to have hydrogen at its natural abundance isotopic composition. Accordingly, in the compounds of this disclosure any atom specifically designated as a 25 deuterium (D) is meant to represent deuterium. The terms “treat,” “treatment,” and “treating” refer to any indicia of success in the treatment or amelioration of an injury, pathology, condition (e.g., cancer), or symptom (e.g., cognitive impairment), including any objective or subjective parameter such as abatement; remission; diminishing of symptoms or making the symptom, injury, pathology, or condition 30 more tolerable to the patient; reduction in the rate of symptom progression; decreasing the frequency or duration of the symptom or condition; or, in some situations, preventing the onset of the symptom. The treatment or amelioration of symptoms can be based on any objective or subjective parameter, including, for example, the result of a physical examination. The terms “cancer,” “neoplasm,” and “tumor” are used herein to refer to cells which 35 exhibit autonomous, unregulated growth, such that the cells exhibit an aberrant growth 20 15077.006WO1 phenotype characterized by a significant loss of control over cell proliferation. Cells of interest for detection, analysis, and/or treatment in the context of the invention include cancer cells (e.g., cancer cells from an individual with cancer), malignant cancer cells, pre-metastatic cancer cells, metastatic cancer cells, and non-metastatic cancer cells. Cancers of virtually every tissue are 5 known. The phrase “cancer burden” refers to the quantum of cancer cells or cancer volume in a subject. Reducing cancer burden accordingly refers to reducing the number of cancer cells or the cancer cell volume in a subject. The term “cancer cell” as used herein refers to any cell that is a cancer cell (e.g., from any of the cancers for which an individual can be treated, e.g., isolated from an individual having cancer) or is derived from a cancer cell, e.g., clone of a cancer cell. 10 For example, a cancer cell can be from an established cancer cell line, can be a primary cell isolated from an individual with cancer, can be a progeny cell from a primary cell isolated from an individual with cancer, and the like. In some embodiments, the term can also refer to a portion of a cancer cell, such as a sub-cellular portion, a cell membrane portion, or a cell lysate of a cancer cell. Many types of cancers are known to those of skill in the art, including solid 15 tumors such as carcinomas, sarcomas, glioblastomas, melanomas, lymphomas, and myelomas, and circulating cancers such as leukemias. As used herein, the term “cancer” includes any form of cancer, including but not limited to, solid tumor cancers (e.g., skin, lung, prostate, breast, gastric, bladder, colon, ovarian, pancreas, kidney, liver, glioblastoma, medulloblastoma, leiomyosarcoma, head & neck 20 squamous cell carcinomas, melanomas, and neuroendocrine) and liquid cancers (e.g., hematological cancers); carcinomas; soft tissue tumors; sarcomas; teratomas; melanomas; leukemias; lymphomas; and brain cancers, including minimal residual disease, and including both primary and metastatic tumors. The phrases “effective amount” and “therapeutically effective amount” refer to a dose or 25 amount of a substance such as an antibody conjugate that produces therapeutic effects for which it is administered. The exact dose will depend on the purpose of the treatment, and will be ascertainable by one skilled in the art using known techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms (vols.1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999); Goodman & 30 Gilman’s The Pharmacological Basis of Therapeutics, 11^th Edition (McGraw-Hill, 2006); and Remington: The Science and Practice of Pharmacy, 22^nd Edition, (Pharmaceutical Press, London, 2012)). In the case of cancer, the therapeutically effective amount of the antibody conjugate may reduce the number of cancer cells; reduce the tumor size; inhibit (i.e., slow to some extent and preferably stop) cancer cell infiltration into peripheral organs; inhibit (i.e., slow 35 to some extent and preferably stop) tumor metastasis; inhibit, to some extent, tumor growth; 21 15077.006WO1 and/or relieve to some extent one or more of the symptoms associated with the cancer. To the extent the antibody conjugate may prevent growth and/or kill existing cancer cells, it may be cytostatic and/or cytotoxic. For cancer therapy, efficacy can, for example, be measured by assessing the time to disease progression (TTP) and/or determining the response rate (RR) 5 “Recipient,” “individual,” “subject,” “host,” and “patient” are used interchangeably and refer to any mammalian subject for whom diagnosis, treatment, or therapy is desired (e.g., humans). “Mammal” for purposes of treatment refers to any animal classified as a mammal, including humans, domestic and farm animals, and zoo, sports, or pet animals, such as dogs, horses, cats, cows, sheep, goats, pigs, camels, etc. In certain embodiments, the mammal is 10 human. As used herein, the term “administering” refers to parenteral, intravenous, intraperitoneal, intramuscular, intratumoral, intralesional, intranasal, or subcutaneous administration, oral administration, administration as a suppository, topical contact, intrathecal administration, or the implantation of a slow-release device, e.g., a mini-osmotic pump, to the 15 subject. The terms “about” and “around,” as used herein to modify a numerical value, indicate a close range surrounding the numerical value. Thus, if “X” is the value, “about X” or “around X” indicates a value of from 0.9X to 1.1X, e.g., from 0.95X to 1.05X or from 0.99X to 1.01X. A reference to “about X” or “around X” specifically indicates at least the values X, 0.95X, 0.96X, 20 0.97X, 0.98X, 0.99X, 1.01X, 1.02X, 1.03X, 1.04X, and 1.05X. Accordingly, “about X” and “around X” are intended to teach and provide written description support for a claim limitation of, e.g., “0.98X.” ANTIBODIES The antibody conjugate compositions of the invention comprise an antibody. Included in 25 the scope of the embodiments of the invention are functional variants of the antibody constructs or antigen binding domain described herein. The term “functional variant” as used herein refers to an antibody construct having an antigen binding domain with substantial or significant sequence identity or similarity to a parent antibody construct or antigen binding domain, which functional variant retains the biological activity of the antibody construct or antigen binding 30 domain of which it is a variant. Functional variants encompass, for example, those variants of the antibody constructs or antigen binding domain described herein (the parent antibody construct or antigen binding domain) that retain the ability to recognize target cells expressing a tumor-associated antigen or cell surface receptor to a similar extent, the same extent, or to a higher extent, as the parent antibody construct or antigen binding domain. 22 15077.006WO1 In reference to the antibody construct or antigen binding domain, the functional variant can, for instance, be at least about 30%, about 50%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or more identical in amino acid sequence to the antibody construct or antigen 5 binding domain. A functional variant can, for example, comprise the amino acid sequence of the parent antibody construct or antigen binding domain with at least one conservative amino acid substitution. Alternatively, or additionally, the functional variants can comprise the amino acid sequence of the parent antibody construct or antigen binding domain with at least one non- 10 conservative amino acid substitution. In this case, it is preferable for the non-conservative amino acid substitution to not interfere with or inhibit the biological activity of the functional variant. The non-conservative amino acid substitution may enhance the biological activity of the functional variant, such that the biological activity of the functional variant is increased as compared to the parent antibody construct or antigen binding domain. 15 A functional variant can, for example, comprise the amino acid sequence of the parent antibody construct or antigen binding with at least one non-canonical amino acid (ncAA) substitution (L Wang, et al, (2001) Science , 292(5516):498-500, CC Liu, PG Schultz, (2010) Annu Rev Biochem.79:413-44). The antibodies comprising the antibody conjugate compositions of the invention include 20 Fc engineered variants. In some embodiments, the mutations in the Fc region that result in modulated binding to one or more Fc receptors can include one or more of the following mutations: YTE (M252Y/S254T/T256E), LALAPA (L234A/L235A/P329A), SD (S239D), SDIE (S239D/I332E), SE (S267E), SELF (S267E/L328F), SDIE (S239D/I332E), SDIEAL (S239D/I332E/A330L), GA (G236A), ALIE (A330L/I332E), GASDALIE 25 (G236A/S239D/A330L/I332E), V9 (G237D/P238D/P271G/A330R), and V11 (G237D/P238D/H268D/P271G/A330R), and/or one or more mutations at the following amino acids: E345R, E345R/E430G, E345K, E233, G237, P238, H268, P271, L328 and A330. Additional Fc region modifications for modulating Fc receptor binding are described in, for example, US 2016/0145350, US 7416726 and US 5624821, which are hereby incorporated by 30 reference in their entireties herein. The antibodies comprising the antibody conjugate compositions of the invention include glycan variants, such as afucosylation. In some embodiments, the Fc region of the binding agents are modified to have an altered glycosylation pattern of the Fc region compared to the native non-modified Fc region. 23 15077.006WO1 In some embodiments, the antibodies in the antibody conjugate compositions contain a modified Fc region, wherein the modification modulates the binding of the Fc region to one or more Fc receptors. In some embodiments, the antibodies in the antibody conjugate contain one or more 5 modifications (e.g., amino acid insertion, deletion, and/or substitution) in the Fc region that results in modulated binding (e.g., increased binding or decreased binding) to one or more Fc receptors (e.g., FcγRI (CD64), FcγRIIA (CD32A), FcγRIIB (CD32B), FcγRIIIA (CD16a), and/or FcγRIIIB (CD16b)) as compared to the native antibody lacking the mutation in the Fc region. In some embodiments, the antibodies in the antibody conjugate compositions contain 10 one or more modifications (e.g., amino acid insertion, deletion, and/or substitution) in the Fc region that reduce the binding of the Fc region of the antibody to FcγRIIB. In some embodiments, the antibodies in the antibody conjugate compositions contain one or more modifications (e.g., amino acid insertion, deletion, and/or substitution) in the Fc region of the antibody that reduce the binding of the antibody to FcγRIIB while maintaining the same binding 15 or having increased binding to FcγRI (CD64), FcγRIIA (CD32A), and/or FcRγIIIA (CD16a) as compared to the native antibody lacking the mutation in the Fc region. In some embodiments, the antibodies in the antibody conjugate compositions contain one of more modifications in the Fc region that increase the binding of the Fc region of the antibody to FcγRIIB. In some embodiments, the modulated binding is provided by mutations in the Fc region 20 of the antibody relative to the native Fc region of the antibody. The mutations can be in a CH2 domain, a CH3 domain, or a combination thereof. A “native Fc region” is synonymous with a “wild-type Fc region” and comprises an amino acid sequence that is identical to the amino acid sequence of an Fc region found in nature or identical to the amino acid sequence of the Fc region found in the native antibody (e.g., cetuximab). Native sequence human Fc regions 25 include a native sequence human IgG1 Fc region, native sequence human IgG2 Fc region, native sequence human IgG3 Fc region, and native sequence human IgG4 Fc region, as well as naturally occurring variants thereof. Native sequence Fc includes the various allotypes of Fcs (Jefferis et al., (2009) mAbs, 1(4):332-338). In some embodiments, the Fc region of the antibodies of the antibody conjugate 30 compositions are modified to have an altered glycosylation pattern of the Fc region compared to the native non-modified Fc region. Human immunoglobulin is glycosylated at the Asn297 residue in the Cγ2 domain of each heavy chain. This N-linked oligosaccharide is composed of a core heptasaccharide, N-acetylglucosamine4Mannose3 (GlcNAc4Man3). Removal of the heptasaccharide with endoglycosidase or PNGase F is known to lead to conformational changes 35 in the antibody Fc region, which can significantly reduce antibody-binding affinity to activating 24 15077.006WO1 FcγR and lead to decreased effector function. The core heptasaccharide is often decorated with galactose, bisecting GlcNAc, fucose, or sialic acid, which differentially impacts Fc binding to activating and inhibitory FcγR. Additionally, it has been demonstrated that α2,6-sialyation enhances anti-inflammatory activity in vivo, while afucosylation leads to improved FcγRIIIa 5 binding and a 10-fold increase in antibody-dependent cellular cytotoxicity and antibody- dependent phagocytosis. Specific glycosylation patterns, therefore, can be used to control inflammatory effector functions. In some embodiments, the modification to alter the glycosylation pattern is a mutation. For example, a substitution at Asn297. In some embodiments, Asn297 is mutated to glutamine10 (N297Q). Methods for controlling immune response with antibodies that modulate FcγR- regulated signaling are described, for example, in US 7416726, US 2007/0014795 and US 2008/0286819, which are hereby incorporated by reference in their entireties. In some embodiments, the antibodies of the antibody conjugate compositions are modified to contain an engineered Fab region with a non-naturally occurring glycosylation 15 pattern. For example, hybridomas can be genetically engineered to secrete afucosylated mAb, desialylated mAb or deglycosylated Fc with specific mutations that enable increased FcRγIIIa binding and effector function. In some embodiments, the antibodies of the antibody conjugate compositions are engineered to be afucosylated or glycosylated. In some embodiments, the antibodies in the antibody conjugate compositions are a 20 cysteine-engineered antibody which provides for site-specific conjugation of an adjuvant, label, or drug moiety to the antibody through cysteine substitutions at sites where the engineered cysteines are available for conjugation but do not perturb immunoglobulin folding and assembly or alter antigen binding and effector functions (Junutula, et al., (2008) Nature Biotech., 26(8):925-932; Dornan et al. (2009) Blood 114(13):2721-2729; US 7521541; US 7723485; US 25 2012/0121615; WO 2009/052249). A “cysteine engineered antibody” or “cysteine engineered antibody variant” is an antibody in which one or more residues of an antibody are substituted with cysteine residues. Cysteine-engineered antibodies can be conjugated to the TPI-VHL moiety with uniform stoichiometry (e.g., up to two TPI-VHL moieties per antibody in an antibody that has a single engineered cysteine site). 30 In some embodiments, cysteine-engineered antibodies are used to prepare antibody conjugate compositions with a reactive cysteine thiol residue introduced at a site on the light chain, such as the 149-lysine site (LC K149C), or on the heavy chain such as the 122-serine site (HC S122C), as numbered by Kabat numbering. In other embodiments, the cysteine-engineered antibodies have a cysteine residue introduced at the 375-serine site (EU numbering) of the heavy 35 chain (HC S375C). In other embodiments, the cysteine-engineered antibodies have a cysteine 25 15077.006WO1 residue introduced at the 118-alanine site (EU numbering) of the heavy chain (HC A118C). This site is alternatively numbered 121 by Sequential numbering or 114 by Kabat numbering. In other embodiments, the cysteine-engineered antibodies have a cysteine residue introduced in: (i) the light chain at G64C, R142C, K188C, L201C, T129C, S114C, or E105C according to Kabat 5 numbering; (ii) the heavy chain at D101C, V184C, T205C, or S122C according to Kabat numbering; or (iii) other cysteine-mutant antibodies, and as described in Bhakta, S. et al, (2013) “Engineering THIOMABs for Site-Specific Conjugation of Thiol-Reactive Linkers”, Laurent Ducry (ed.), Antibody-Drug Conjugates, Methods in Molecular Biology, vol.1045, pages 189- 203; WO 2011/156328; US 9000130. 10 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, the antibody is an anti-CD40 antibody, an antibody selected from an anti-LRRC15 antibody, an anti-CTSK antibody, an anti-ADAM12 antibody, an anti-ITGA1115 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-Nectin4 antibody, an anti- 20 ASGR1 antibody, and an anti-MUC16 antibody. In some embodiments, the antibody or Fc fusion protein is selected from: abagovomab, abatacept (also known as ORENCIA®), abciximab (also known as REOPRO®), c7E3 Fab), adalimumab (also known as HUMIRA®), adecatumumab, alemtuzumab (also known as CAMPATH®), MabCampath or Campath-1H), altumomab, afelimomab, panitumumab, 25 mafenatox, anrukizumab, apolizumab, arcitumomab, aselizumab, atlizumab, atorolimumab, bapineuzumab, basiliximab (also known as SIMULECT®), bavituximab, bectumomab (also known as LYMPHOSCAN®), belimumab (also known as LYMPHO-STAT-B®), bertilimumab, besilesomab, bevacizumab (also known as AVASTIN®), biciromab brallobarbital, bivatuzumab mertansine, campath, canakinumab (also known as ACZ885), cantuzumab mertansine, capromab 30 (also known as PROSTASCINT®), catumaxomab (also known as REMOVAB®), cedelizumab (also known as CIMZIA®), certolizumab pegol, cetuximab (also known as ERBITUX®), clenoliximab, dacetuzumab, dacliximab, daclizumab (also known as ZENAPAX®), datopotamab, denosumab (also known as AMG 162), detumomab, dorlimomab aritox, dorlixizumab, duntumumab, durimulumab, durmulumab, ecromeximab, eculizumab (also 35 known as SOLIRIS®), edobacomab, edrecolomab (also known as Mab17-1A, PANOREX®), 26 15077.006WO1 efalizumab (also known as RAPTIVA®), efungumab (also known as MYCOGRAB®), elsilimomab, enapotamab, enfortumab, enlimomab pegol, epitumomab cituxetan, efalizumab, epitumomab, epratuzumab, erlizumab, ertumaxomab (also known as REXOMUN®), etanercept (also known as ENBREL®), etaracizumab (also known as etaratuzumab, VITAXIN®, 5 ABEGRIN®), exbivirumab, fanolesomab (also known as NEUTROSPEC®), faralimomab, felvizumab, fontolizumab (also known as HUZAF®), galiximab, gantenerumab, gavilimomab (also known as ABXCBL®), gemtuzumab ozogamicin (also known as MYLOTARG®), golimumab (also known as CNTO 148), gomiliximab, ibalizumab (also known as TNX-355), ibritumomab tiuxetan (also known as ZEVALIN®), igovomab, imciromab, indusatumab, 10 infliximab (also known as REMICADE®), inolimomab, inotuzumab ozogamicin, ipilimumab (also known as MDX-010, MDX-101), iratumumab, keliximab, labetuzumab, ladiratuzumab, lemalesomab, lebrilizumab, lerdelimumab, lexatumumab (also known as, HGS-ETR2, ETR2- ST01), lexitumumab, libivirumab, lintuzumab, loncastuximab, losatuxizumab, lucatumumab, lumiliximab, mapatumumab (also known as HGSETR1, TRM-1), maslimomab, matuzumab 15 (also known as EMD72000), mepolizumab (also known as BOSATRIA®), metelimumab, milatuzumab, miltuximab, minretumomab, mirvetuximab, mitumomab, morolimumab, motavizwnab (also known as NUMAX®), muromonab (also known as OKT3), nacolomab tafenatox, naptumomab estafenatox, natalizumab (also known as TYSABRI®, ANTEGREN®), nebacumab, nerelimomab, nimotuzumab (also known as THERACIM hR3®, THERA-CIM- 20 hR3®, THERALOC®), nofetumomab merpentan (also known as VERLUMA®), ocrelizumab, odulimomab, ofatumumab, omalizumab (also known as XOLAIR®), oregovomab (also known as OVAREX®), otelixizumab, pagibaximab, palivizumab (also known as SYNAGIS®), panitumumab (also known as ABX-EGF, VECTIBIX®), pascolizumab, pemtumomab (also known as THERAGYN®), pertuzumab (also known as 2C4, OMNITARG®), pexelizumab, 25 pinatuzumab, pintumomab, polatuzumab, priliximab, pritumumab, ranibizumab (also known as LUCENTIS®), raxibacumab, regavirumab, reslizumab, rituximab (also known as RITUXAN®, MabTHERA®), rovelizumab, ruplizumab, sacituzumab, satumomab, sevirumab, sibrotuzumab, siplizumab (also known as MEDI-507), sontuzumab, stamulumab (also known as MYO-029), sulesomab (also known as LEUKOSCAN®), tacatuzumab tetraxetan, tadocizumab, talizumab, 30 taplitumomab paptox, tefibazumab (also known as AUREXIS®), telimomab aritox, teneliximab, teplizumab, ticilimumab, tocilizumab (also known as ACTEMRA®), tisotumab, toralizumab, tositumomab, trastuzumab (also known as HERCEPTIN®), tremelimumab (also known as CP- 675,206), tucotuzumab celmoleukin, tusamitamab, tuvirumab, urtoxazumab, ustekinumab (also known as CNTO 1275), vapaliximab, veltuzumab, vepalimomab, visilizumab (also known as 35 NUVION®), volociximab (also known as M200), votumumab (also known as 27 15077.006WO1 HUMASPECT®), zalutumumab, zanolimumab (also known as HuMAX-CD4), ziralimumab, zolimomab aritox, daratumumab, elotuxumab, obintunzumab, olaratumab, brentuximab vedotin, afibercept, abatacept, belatacept, afibercept, etanercept, romiplostim, SBT-040 (sequences listed in US 2017/0158772. In some embodiments, the antibody is rituximab. 5 In an exemplary embodiment, the antibody conjugate composition of the invention comprises an antibody construct that comprises an antigen binding domain that specifically recognizes and binds HER2. In certain embodiments, the antibody conjugate composition comprises an anti-HER2 antibody. In one embodiment of the invention, an anti-HER2 antibody of an antibody conjugate composition of the invention comprises a humanized anti-HER2 10 antibody, e.g., huMAb4D5-1, huMAb4D5-2, huMAb4D5-3, huMAb4D5-4, huMAb4D5-5, huMAb4D5-6, huMAb4D5-7 and huMAb4D5-8, as described in Table 3 of US 5821337, which is specifically incorporated by reference herein. Those antibodies contain human framework regions with the complementarity-determining regions of a murine antibody (4D5) that binds to HER2. The humanized antibody huMAb4D5-8 is also referred to as trastuzumab, commercially15 available under the tradename HERCEPTIN™ (Genentech, Inc.). Trastuzumab (CAS 180288- 69-1, HERCEPTIN^, huMAb4D5-8, rhuMAb HER2, Genentech) is a recombinant DNA- derived, IgG1 kappa, monoclonal antibody that is a humanized version of a murine anti-HER2 antibody (4D5) that selectively binds with high affinity in a cell-based assay (Kd = 5 nM) to the extracellular domain of HER2 (US 5677171; US 5821337; US 6054297; US 6165464; US 20 6339142; US 6407213; US 6639055; US 6719971; US 6800738; US 7074404; Coussens et al (1985) Science 230:1132-9; Slamon et al (1989) Science 244:707-12; Slamon et al (2001) New Engl. J. Med.344:783-792). In an embodiment of the invention, the antibody construct or antigen binding domain comprises the CDR regions of trastuzumab. In an embodiment of the invention, the anti-HER2 25 antibody further comprises the framework regions of the trastuzumab. In an embodiment of the invention, the anti-HER2 antibody further comprises one or both variable regions of trastuzumab. In another embodiment of the invention, an anti-HER2 antibody of an antibody conjugate composition of the invention comprises a humanized anti-HER2 antibody, e.g., 30 humanized 2C4, as described in US 7862817. An exemplary humanized 2C4 antibody is pertuzumab (CAS Reg. No.380610-27-5), PERJETA™ (Genentech, Inc.). Pertuzumab is a HER dimerization inhibitor (HDI) and functions to inhibit the ability of HER2 to form active heterodimers or homodimers with other HER receptors (such as EGFR/HER1, HER2, HER3 and HER4). See, for example, Harari and Yarden, Oncogene 19:6102-14 (2000); Yarden and 35 Sliwkowski. Nat Rev Mol Cell Biol 2:127-37 (2001); Sliwkowski Nat Struct Biol 10:158-9 28 15077.006WO1 (2003); Cho et al. Nature 421:756-60 (2003); and Malik et al. Pro Am Soc Cancer Res 44:176-7 (2003). PERJETA™ is approved for the treatment of breast cancer. In an embodiment of the invention, the antibody construct or antigen binding domain comprises the CDR regions of pertuzumab. In an embodiment of the invention, the anti-HER2 5 antibody further comprises the framework regions of the pertuzumab. In an embodiment of the invention, the anti-HER2 antibody further comprises one or both variable regions of pertuzumab. ANTIBODY TARGETS In some embodiments, the antibody of an antibody conjugate composition is capable of 10 binding one or more targets selected from (e.g., specifically binds to a target selected from) 5T4, ABL, ABCF1, ACVR1, ACVR1B, ACVR2, ACVR2B, ACVRL1, ADORA2A, Aggrecan, AGR2, AICDA, AIF1, AIGI, AKAP1, AKAP2, AMH, AMHR2, ANGPT1, ANGPT2, ANGPTL3, ANGPTL4, ANPEP, APC, APOC1, AR, aromatase, ATX, AX1, Axl, AZGP1 (zinc- a-glycoprotein), B7.1, B7.2, B7-H1, B7-H3, B7-H4, BAD, BAFF, BAG1, BAI1, BCR, BCL2, 15 BCL6, BDNF, BLNK, BLR1 (MDR15), BIyS, BMP1, BMP2, BMP3B (GDFIO), BMP4, BMP6, BMP8, BMPRTA, BMPR1B, BMPR2, BPAG1 (plectin), BRCA1, C19orflO (IL27w), C3, C4A, C5, C5R1, CANT1, CAPRIN-1, CASP1, CASP4, CAV1, CCBP2 (D6/JAB61), CCLI (1-309), CCLI1 (eotaxin), CCL13 (MCP-4), CCL15 (MIP-Id), CCL16 (HCC-4), CCL17 (TARC), CCL18 (PARC), CCL19 (MIP-3b), CCL2 (MCP-1), MCAF, CCL20 (MIP-3a), CCL21 20 (MEP-2), SLC, exodus-2, CCL22(MDC/STC-1), CCL23 (MPIF-I), CCL24 (MPIF-2/eotaxin-2), CCL25 (TECK), CCL26 (eotaxin-3), CCL27 (CTACK/ILC), CCL28, CCL3 (MIP-Ia), CCL4 (MIPIb), CCL5 (RANTES), CCL7 (MCP-3), CCL8 (mcp-2), CCNA1, CCNA2, CCND1, CCNE1, CCNE2, CCR1 (CKR1/HM145), CCR2 (mcp-IRB/RA), CCR3 (CKR3/CMKBR3), CCR4, CCR5 (CMKBR5/ChemR13), CCR6 (CMKBR6/CKR-L3/STRL22/DRY6), CCR7 25 (CKR7/EBI1), CCR8 (CMKBR8/TERI/CKR-L1), CCR9 (GPR-9-6), CCRL1 (VSHK1), CCRL2 (L-CCR), CD164, CD19, CDIC, CD2, CD20, CD21, CD200, CD22, CD24, CD27, CD28, CD3, CD30, CD33, CD35, CD37, CD38, CD3E, CD3G, CD3Z, CD4, CD38, CD40, CD40L, CD44, CD45RB, CD47, CD52, CD69, CD72, CD74, CD79A, CD79B, CD8, CD80, CD81, CD83, CD86, CD117 (cKit), CD123, CD137, CD152, CD274, CDH1 (Ecadherin), CDH3 (Pcadherin), 30 CDH1O, CDH12, CDH13, CDH18, CDH19, CDH2O, CDH5, CDH7, CDH8, CDH9, CDK2, CDK3, CDK4, CDK5, CDK6, CDK7, CDK9, CDKN1A (p21Wap1/Cip1), CDKN1B (p27Kip1), CDKN1C, CDKN2A (p16INK4a), CDKN2B, CDKN2C, CDKN3, CEACAM5, CEACAM6, CEBPB, CERI, CHGA, CHGB, Chitinase, CHST1O, CKLFSF2, CKLFSF3, CKLFSF4, CKLFSF5, CKLFSF6, CKLFSF7, CKLFSF8, CLDN3, CLDN7 (claudin-7), CLDN18.2 29 15077.006WO1 (claudin 18.2), CLL-1, CLN3, CLU (clusterin), cMet, CMKLR1, CMKOR1 (RDC1), CNR1, COL18A1, COLIA1, COL4A3, COL6A1, CR2, Cripto, CRP, CSF1 (M-CSF), CSF2 (GM-CSF), CSF3 (GCSF), CTL8, CTNNB1 (b-catenin), CTSB (cathepsin B), CX3CL1 (SCYD1), CX3CR1 (V28), CXCL1 (GRO1), CXCL1O (IP-IO), CXCLI1 (1-TAC/IP-9), CXCL12 (SDF1), CXCL13, 5 CXCL14, CXCL16, CXCL2 (GRO2), CXCL3 (GRO3), CXCL5 (ENA-78/LIX), CXCL6 (GCP- 2), CXCL9 (MIG), CXCR3 (GPR9/CKR-L2), CXCR4, CXCR6 (TYMSTR/STRL33/Bonzo), CYB5, CYC1, CYSLTR1, DAB2IP, DES, DKFZp451J0118, DNCL1, DPP4, E2F1, Engel, Edge, Fennel, EFNA3, EFNB2, EGF, EGFR, ELAC2, ENG, Enola, ENO2, ENO3, EPHA1, EPHA2, EPHA3, EPHA4, EPHA5, EPHA6, EPHA7, EPHA8, EPHA9, EPRA10, EPHB1,10 EPHB2, EPHB3, EPHB4, EPHB5, EPHB6, EPHRIN-A1, EPHRIN-A2, EPHRINA3, EPHRIN- A4, EPHRIN-A5, EPHRIN-A6, EPHRIN-B1, EPHRIN-B2, EPHRIN-B3, EPHB4, EPG, ERBB2 (Her-2), EREG, ERK8, Estrogen receptor, Earl, ESR2, F3 (TF), FADD, farnesyltransferase, FasL, FAP, FASNf, FCER1A, FCER2, FCGR3A, FGF, FGF1 (aFGF), FGF10, FGF11, FGF12, FGF12B, FGF13, FGF14, FGF16, FGF17, FGF18, FGF19, FGF2 15 (bFGF). FGF20, FGF21, FGF22, FGF23, FGF3 (int-2), FGF4 (HST), FGF5, FGF6 (HST-2), FGF7 (KGF), FGF8, FGF9, FGFR3, FIGF (VEGFD), FILI (EPSILON), FBL1 (ZETA), FLJ12584, FLJ25530, FLRT1 (fibronectin), FLT1, FLT-3, FOLR1 (folate receptor alpha), FOS, FOSL1 (FRA-1), FY (DARC), GABRP (GABAa), GAGEB1, GAGEC1, GALNAC4S-6ST, GATA3, GD2, GDF5, GFI1, GGT1, GM-CSF, GNAS1, GNRH1, GPC-1, GPR2 (CCR10), 20 GPR31, GPR44, GPR81 (FKSG80), GRCC1O (C1O), GRP, GSN (Gelsolin), GSTP1, HAVCR2, HDAC, HDAC4, HDAC5, HDAC7A, HDAC9, Hedgehog, HGF, HIF1A, HIP1, histamine and histamine receptors, HLA-A, HLA-DRA, HLA-E, HM74, HMOXI, HSP90, HUMCYT2A, ICEBERG, ICOSL, ID2, IFN-a, IFNA1, IFNA2, IFNA4, IFNA5, EFNA6, BFNA7, IFNB1, IFNgamma, IFNW1, IGBP1, IGF1, IGFIR, IGF2, IGFBP2, IGFBP3, IGFBP6, DL-1, ILIO, 25 ILIORA, ILIORB, IL-1, IL1R1 (CD121a), IL1R2 (CD121b), IL-IRA, IL-2, IL2RA (CD25), IL2RB (CD122), IL2RG (CD132), IL-4, IL-4R (CD123), IL-5, IL5RA (CD125), IL3RB (CD131), IL-6, IL6RA, (CD126), IR6RB (CD130), IL-7, IL7RA (CD127), IL-8, CXCR1 (IL8RA), CXCR2, (IL8RB/CD128), IL-9, IL9R (CD129), IL-10, IL10RA (CD210), IL10RB (CDW210B), IL-11, IL11RA, IL-12, IL-12A, IL-12B, IL-12RB1, IL-12RB2, IL-13, IL13RA1, 30 IL13RA2, IL14, IL15, IL15RA, IL16, IL17, IL17A, IL17B, IL17C, IL17R, IL18, IL18BP, IL18R1, IL18RAP, IL19, ILIA, ILIB, ILIF10, ILIF5, IL1F6, ILIF7, IL1F8, DL1F9, ILIHYI, ILIR1, ILIR2, ILIRAP, ILIRAPLI, ILIRAPL2, ILIRL1, IL1RL2, ILIRN, IL2, IL20, IL20RA, IL21R, IL22, IL22R, IL22RA2, IL23, DL24, IL25, IL26, IL27, IL28A, IL28B, IL29, IL2RA, IL2RB, IL2RG, IL3, IL30, IL3RA, IL4, IL4, IL6ST (glycoprotein 130), ILK, INHA, INHBA, 35 INSL3, INSL4, IRAK1, IRAK2, ITGA1, ITGA2, ITGA3, ITGA6 (.alpha.6 integrin), ITGAV, 30 15077.006WO1 ITGB3, ITGB4 (.beta.4 integrin), ITGB6 (beta.6 integrin), JAG1, JAK1, JAK3, JTB, JUN, K6HF, KAI1, KDR, KITLG, KLF5 (GC Box BP), KLF6, KLK10, KLK12, KLK13, KLK14, KLK15, KLK3, KLK4, KLK5, KLK6, KLK9, KRT1, KRT19 (Keratin 19), KRT2A, KRTHB6 (hair-specific type II keratin), LAMA5, LEP (leptin), Lingo-p75, Lingo-Troy, LIV-1, LPS, 5 LRRC15, LTA (TNF-b)), LTB, LTB4R (GPR16), LTB4R2, LTBR, MACMARCKS, MAG or OMgp, MAP2K7 (c-Jun), MCP-1, MDK, MIB1, midkine, MIF, MISRII, MJP-2, MK, MKI67 (Ki-67), MMP2, MMP9, MS4A1, MSMB, MT3 (metallothionectin-UI), mTOR, MTSS1, MUC1 (mucin), MUC16, MYC, MYD88, NCK2, neurocan, Nectin-4, NFKBI, NFKB2, NGFB (NGF), NGFR, NgR-Lingo, NgRNogo66, (Nogo), NgR-p75, NgR-Troy, NMEI (NM23A), NOTCH, 10 NOTCH1, NOX5, NPPB, NROB1, NROB2, NRID1, NR1D2, NR1H2, NR1H3, NR1H4, NR112, NR113, NR2C1, NR2C2, NR2E1, NR2E3, NR2F1, NR2F2, NR2F6, NR3C1, NR3C2, NR4A1, NR4A2, NR4A3, NR5A1, NR5A2, NR6A1, NRP1, NRP2, NT5E, NTN4, ODZI, OPRDI, P2RX7, PAP, PART1, PATE, PAWR, PCA3, PCDGF, PCNA, PDGFA, PDGFB, PDGFRA, PDGFRB, PECAMI, peg-asparaginase, PF4 (CXCL4), PGF, PGR, phosphacan, 15 PIAS2, PI3 Kinase, PIK3CG, PLAU (uPA), PLG, PLXDCI, PKC, PKC-beta, PPBP (CXCL7), PPID, PR1, PRKCQ, PRKD1, PRL, PROC, PROK2, PSAP, PSCA, PSMA, PTAFR, PTEN, PTGS2 (COX-2), PIN, RAC2 (P21Rac2), RANK, RANK ligand, RARB, RGS1, RGS13, RGS3, RNFI1O (ZNF144), Ron, ROBO2, RXR, S100A2, SCGB 1D2 (lipophilin B), SCGB2A1 (mammaglobin 2), SCGB2A2 (mammaglobin 1), SCYE1 (endothelial Monocyte-activating 20 cytokine), SDF2, SERPENA1, SERPINA3, SERPINB5 (maspin), SERPINEI (PAI-I), SERPINFI, SHIP-1, SHIP-2, SHB1, SHB2, SHBG, SfcAZ, SLC2A2, SLC33A1, SLC43A1, SLIT2, SPP1, SPRR1B (Spr1), ST6GAL1, STAB1, STATE, STEAP, STEAP2, TB4R2, TBX21, TCP1O, TDGF1, TEK, TGFA, TGFB1, TGFB1I1, TGFB2, TGFB3, TGFBI, TGEBR1, TGFBR2, TGFBR3, THIL, THBS1 (thrombospondin-1), THBS2, THBS4, THPO, TIE (Tie-1), 25 TIMP3, tissue factor, TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10, TLR11, TNF, TNF-a, TNFAIP2 (B94), TNFAIP3, TNFRSF11A, TNFRSF1A, TNFRSF1B, TNFRSF21, TNFRSF5, TNFRSF6 (Fas), TNFRSF7, TNFRSF8, TNFRSF9, TNFSF1O (TRAIL), TNFSF11 (TRANCE), TNFSF12 (APO3L), TNFSF13 (April), TNFSF13B, TNSF14 (HVEM-L), TNFRSF14 (HVEM), TNFSF15 (VEGI), TNFSF18, TNFSF4 (OX40 ligand), 30 TNFSF5 (CD40 ligand). TNFSF6 (FasL), TNFSF7 (CD27 ligand), TNFSF8 (CD30 ligand), TNFSF9 (4-1BB ligand), TOLLIP, Toll-like receptors, TOP2A (topoisomerase 1ia), TP53, TPM1, TPM2, TRADD, TRAF1, TRAF2, TRAF3, TRAF4, TRAF5, TRAF6, TRKA, TREM1, TREM2, TROP2, TRPC6, TSLP, TWEAK, Tyrosinase, uPAR, VEGF, VEGFB, VEGFC, versican, VHL C5, VLA-4, Wnt-1, XCL1 (tymphotactin), XCL2 (SCM-Ib), XCRI 35 (GPR5/CCXCR1), YYI, ZFPM2, CLEC4C (BDCA-2, DLEC, CD303, CLECSF7), CLEC4D 31 15077.006WO1 (MCL, CLECSF8), CLEC4E (Mincle), CLEC6A (Dectin-2). CLEC5A (MDL-1, CLECSF5), CLEC1B (CLEC-2), CLEC9A (DNGR-1), CLEC7A (Dectin-1), PDGFRa, SLAMF7, GP6 (GPVI), LILRA1 (CD85I), LILRA2 (CD85H, ILT1), LILRA4 (CD85G, ILT7), LILRA5 (CD85F, ILT11), LILRA6 (CD85b, ILT8), NCR1 (CD335, LY94, NKp46), NCR3 (CD335, 5 LY94, NKp46), NCR3 (CD337, NKp30), OSCAR, TARM1, CD300C, CD300E, CD300LB (CD300B), CD300LD (CD300D), KIR2DL4 (CD158D), KIR2DS, KLRC2 (CD159C, NKG2C), KLRK1 (CD314, NKG2D), NCR2 (CD336, NKp44), PILRB, SIGLEC1 (CD169, SN), SIGLEC14, SIGLEC15 (CD33L3), SIGLEC16, SIRPB1 (CD172B), TREM1 (CD354), Tissue Factor (CD142), TREM2, and KLRF1 (NKp80). 10 In some embodiments, the antibody binds to an antigen selected from CDH1, CD19, CD20, CD29, CD30, CD38, CD40, CD47, EpCAM, MUC1, MUC16, EGFR, HER2, SLAMF7, and gp75. In some embodiments, the antibody of an antibody conjugate composition of the invention is capable of binding to one or more tumor-associated antigens (TAA), cell-surface 15 receptors, and immune-specific antigens to confer specificity to the targeting of the conjugate and enable safe and systemic delivery of an active drug moiety. Certain tumor-associated antigens are known in the art, and can be prepared for use in generating antibodies using methods and information which are well known in the art. In attempts to discover effective cellular targets for cancer diagnosis and therapy, researchers have 20 sought to identify transmembrane or otherwise tumor-associated polypeptides that are specifically expressed on the surface of one or more particular type(s) of cancer cell as compared to on one or more normal non-cancerous cell(s). Often, such tumor-associated polypeptides are more abundantly expressed on the surface of the cancer cells as compared to on the surface of the non-cancerous cells. The identification of such tumor-associated cell surface 25 antigen polypeptides allows more specificity in targeting cancer cells for destruction via antibody-based therapies. Examples of TAAs include, but are not limited to, those listed below including (1)-(54). For convenience, information relating to these antigens, all of which are known in the art, is listed below and includes names, alternative names, Genbank accession numbers and primary 30 reference(s), following nucleic acid and protein sequence identification conventions of the National Center for Biotechnology Information (NCBI). Nucleic acid and protein sequences corresponding to TAAs listed below including (1)-(54) are available in public databases such as GenBank. TAAs targeted by antibodies include all amino acid sequence variants and isoforms possessing at least about 70%, 80%, 85%, 90%, or 95% sequence identity relative to the 35 sequences identified in the cited references, and/or which exhibit substantially the same 32 15077.006WO1 biological properties or characteristics as a TAA having a sequence found in the cited references. For example, a TAA having a variant sequence generally is able to bind specifically to an antibody that binds specifically to the TAA with the corresponding sequence listed. The sequences and disclosure in the reference specifically recited herein are expressly incorporated 5 by reference. The disclosure in the references specifically recited herein are expressly incorporated by reference. (1) BMPR1B (bone morphogenetic protein receptor-type IB, Genbank accession no. NM_001203) ten Dijke, P., et al. Science 264 (5155):101-104 (1994), Oncogene 14 (11):1377- 10 1382 (1997)); WO2004063362 (Claim 2); WO2003042661 (Claim 12); US2003134790-A1 (Page 38-39); WO2002102235 (Claim 13; Page 296); WO2003055443 (Page 91-92); WO200299122 (Example 2; Page 528-530); WO2003029421 (Claim 6); WO2003024392 (Claim 2; Fig 112); WO200298358 (Claim 1; Page 183); WO200254940 (Page 100-101); WO200259377(Page 349-350); WO200230268 (Claim 27; Page 376); WO200148204 15 (Example; Fig 4) NP_001194 bone morphogenetic protein receptor, type IB /pid=NP_001194.1 – Cross-references: MIM:603248; NP_001194.1; AY065994. (2) E16 (LAT1, SLC7A5, Genbank accession no. NM_003486) Biochem. Biophys. Res. Commun.255 (2), 283-288 (1999), Nature 395 (6699):288-291 (1998), Gaugitsch, H.W., et al. (1992) J. Biol. Chem.267 (16):11267-11273); WO2004048938 (Example 2); WO2004032842 20 (Example IV); WO2003042661 (Claim 12); WO2003016475 (Claim 1); WO200278524 (Example 2); WO200299074 (Claim 19; Page 127-129); WO200286443 (Claim 27; Pages 222, 393); WO2003003906 (Claim 10; Page 293); WO200264798 (Claim 33; Page 93-95); WO200014228 (Claim 5; Page 133-136); US2003224454 (Fig 3); WO2003025138 (Claim 12; Page 150); NP_003477 solute carrier family 7 (cationic amino acid transporter, y+ system), 25 member 5 /pid=NP_003477.3 – Homo sapiens Cross-references: MIM:600182; NP_003477.3; NM_015923; NM_003486_1. (3) STEAP1 (six transmembrane epithelial antigen of prostate, Genbank accession no. NM_012449) Cancer Res.61 (15), 5857-5860 (2001), Hubert, R.S., et al. (1999) Proc. Natl. Acad. Sci. U.S.A.96 (25):14523-14528); WO2004065577 (Claim 6); WO2004027049 (Fig 1L); 30 EP1394274 (Example 11); WO2004016225 (Claim 2); WO2003042661 (Claim 12); US2003157089 (Example 5); US2003185830 (Example 5); US2003064397 (Fig 2); WO200289747 (Example 5; Page 618-619); WO2003022995 (Example 9; Fig 13A, Example 53; Page 173, Example 2; Fig 2A); NP_036581 six transmembrane epithelial antigen of the prostate Cross-references: MIM:604415; NP_036581.1; NM_012449_1. 33 15077.006WO1 (4) 0772P (CA125, MUC16, Genbank accession no. AF361486) J. Biol. Chem.276 (29):27371-27375 (2001)); WO2004045553 (Claim 14); WO200292836 (Claim 6; Fig 12); WO200283866 (Claim 15; Page 116-121); US2003124140 (Example 16); US 798959. Cross- references: GI:34501467; AAK74120.3; AF361486_1. 5 (5) MPF (MPF, MSLN, SMR, megakaryocyte potentiating factor, mesothelin, Genbank accession no. NM_005823) Yamaguchi, N., et al. Biol. Chem.269 (2), 805-808 (1994), Proc. Natl. Acad. Sci. U.S.A.96 (20):11531-11536 (1999), Proc. Natl. Acad. Sci. U.S.A.93 (1):136- 140 (1996), J. Biol. Chem.270 (37):21984-21990 (1995)); WO2003101283 (Claim 14); (WO2002102235 (Claim 13; Page 287-288); WO2002101075 (Claim 4; Page 308-309); 10 WO200271928 (Page 320-321); WO9410312 (Page 52-57); Cross-references: MIM:601051; NP_005814.2; NM_005823_1. (6) Napi3b (NAPI-3B, NPTIIb, SLC34A2, solute carrier family 34 (sodium phosphate), member 2, type II sodium-dependent phosphate transporter 3b,Genbank accession no. NM_006424) J. Biol. Chem.277 (22):19665-19672 (2002), Genomics 62 (2):281-284 (1999), 15 Field, J.A., et al. (1999) Biochem. Biophys. Res. Commun.258 (3):578-582); WO2004022778 (Claim 2); EP1394274 (Example 11); WO2002102235 (Claim 13; Page 326); EP875569 (Claim 1; Page 17-19); WO200157188 (Claim 20; Page 329); WO2004032842 (Example IV); WO200175177 (Claim 24; Page 139-140); Cross-references: MIM:604217; NP_006415.1; NM_006424_1. 20 (7) Sema 5b (FLJ10372, KIAA1445, Mm.42015, SEMA5B, SEMAG, Semaphorin 5b Hlog, sema domain, seven thrombospondin repeats (type 1 and type 1-like), transmembrane domain (TM) and short cytoplasmic domain, (34emaphoring) 5B, Genbank accession no. AB040878) Nagase T., et al. (2000) DNA Res.7 (2):143-150); WO2004000997 (Claim 1); WO2003003984 (Claim 1); WO200206339 (Claim 1; Page 50); WO200188133 (Claim 1; Page 25 41-43, 48-58); WO2003054152 (Claim 20); WO2003101400 (Claim 11); Accession: Q9P283; EMBL; AB040878; BAA95969.1. Genew; HGNC:10737. (8) PSCA hlg (2700050C12Rik, C530008O16Rik, RIKEN cDNA 2700050C12, RIKEN cDNA 2700050C12 gene, Genbank accession no. AY358628); Ross et al. (2002) Cancer Res. 62:2546-2553; US2003129192 (Claim 2); US2004044180 (Claim 12); US2004044179 (Claim 30 11); US2003096961 (Claim 11); US2003232056 (Example 5); WO2003105758 (Claim 12); US2003206918 (Example 5); EP1347046 (Claim 1); WO2003025148 (Claim 20); Cross- references: GI:37182378; AAQ88991.1; AY358628_1. (9) ETBR (Endothelin type B receptor, Genbank accession no. AY275463); Nakamuta M., et al. Biochem. Biophys. Res. Commun.177, 34-39, 1991; Ogawa Y., et al. Biochem. 35 Biophys. Res. Commun.178, 248-255, 1991; Arai H., et al. Jpn. Circ. J.56, 1303-1307, 1992; 34 15077.006WO1 Arai H., et al. J. Biol. Chem.268, 3463-3470, 1993; Sakamoto A., Yanagisawa M., et al. Biochem. Biophys. Res. Commun.178, 656-663, 1991; Elshourbagy N.A., et al. J. Biol. Chem. 268, 3873-3879, 1993; Haendler B., et al. J. Cardiovasc. Pharmacol.20, s1-S4, 1992; Tsutsumi M., et al. Gene 228, 43-49, 1999; Strausberg R.L., et al. Proc. Natl. Acad. Sci. U.S.A.99, 16899- 5 16903, 2002; Bourgeois C., et al. J. Clin. Endocrinol. Metab.82, 3116-3123, 1997; Okamoto Y., et al. Biol. Chem.272, 21589-21596, 1997; Verheij J.B., et al. Am. J. Med. Genet.108, 223-225, 2002; Hofstra R.M.W., et al. Eur. J. Hum. Genet.5, 180-185, 1997; Puffenberger E.G., et al. Cell 79, 1257-1266, 1994; Attie T., et al., Hum. Mol. Genet.4, 2407-2409, 1995; Auricchio A., et al. Hum. Mol. Genet.5:351-354, 1996; Amiel J., et al. Hum. Mol. Genet.5, 355-357, 1996; 10 Hofstra R.M.W., et al. Nat. Genet.12, 445-447, 1996; Svensson P.J., et al. Hum. Genet.103, 145-148, 1998; Fuchs S., et al. Mol. Med.7, 115-124, 2001; Pingault V., et al. (2002) Hum. Genet.111, 198-206; WO2004045516 (Claim 1); WO2004048938 (Example 2); WO2004040000 (Claim 151); WO2003087768 (Claim 1); WO2003016475 (Claim 1); WO2003016475 (Claim 1); WO200261087 (Fig 1); WO2003016494 (Fig 6); WO2003025138 15 (Claim 12; Page 144); WO200198351 (Claim 1; Page 124-125); EP522868 (Claim 8; Fig 2); WO200177172 (Claim 1; Page 297-299); US2003109676; US6518404 (Fig 3); US5773223 (Claim 1a; Col 31-34); WO2004001004. (10) MSG783 (RNF124, hypothetical protein FLJ20315, Genbank accession no. NM_017763); WO2003104275 (Claim 1); WO2004046342 (Example 2); WO2003042661 20 (Claim 12); WO2003083074 (Claim 14; Page 61); WO2003018621 (Claim 1); WO2003024392 (Claim 2; Fig 93); WO200166689 (Example 6); Cross-references: LocusID:54894; NP_060233.2; NM_017763_1. (11) STEAP2 (HGNC_8639, IPCA-1, PCANAP1, STAMP1, STEAP2, STMP, prostate cancer associated gene 1, prostate cancer associated protein 1, six transmembrane epithelial 25 antigen of prostate 2, six transmembrane prostate protein, Genbank accession no. AF455138) Lab. Invest.82 (11):1573-1582 (2002)); WO2003087306; US2003064397 (Claim 1; Fig 1); WO200272596 (Claim 13; Page 54-55); WO200172962 (Claim 1; Fig 4B); WO2003104270 (Claim 11); WO2003104270 (Claim 16); US2004005598 (Claim 22); WO2003042661 (Claim 12); US2003060612 (Claim 12; Fig 10); WO200226822 (Claim 23; Fig 2); WO200216429 30 (Claim 12; Fig 10); Cross-references: GI:22655488; AAN04080.1; AF455138_1. (12) TrpM4 (BR22450, FLJ20041, TRPM4, TRPM4B, transient receptor potential cation channel, subfamily M, member 4, Genbank accession no. NM_017636) Xu, X.Z., et al. Proc. Natl. Acad. Sci. U.S.A.98 (19):10692-10697 (2001), Cell 109 (3):397-407 (2002), J. Biol. Chem.278 (33):30813-30820 (2003)); US2003143557 (Claim 4); WO200040614 (Claim 14; 35 Page 100-103); WO200210382 (Claim 1; Fig 9A); WO2003042661 (Claim 12); WO200230268 35 15077.006WO1 (Claim 27; Page 391); US2003219806 (Claim 4); WO200162794 (Claim 14; Fig 1A-D); Cross- references: MIM:606936; NP_060106.2; NM_017636_1. (13) CRIPTO (CR, CR1, CRGF, CRIPTO, TDGF1, teratocarcinoma-derived growth factor, Genbank accession no. NP_003203 or NM_003212) Ciccodicola, A., et al. EMBO J.8 5 (7):1987-1991 (1989), Am. J. Hum. Genet.49 (3):555-565 (1991)); US2003224411 (Claim 1); WO2003083041 (Example 1); WO2003034984 (Claim 12); WO200288170 (Claim 2; Page 52- 53); WO2003024392 (Claim 2; Fig 58); WO200216413 (Claim 1; Page 94-95, 105); WO200222808 (Claim 2; Fig 1); US5854399 (Example 2; Col 17-18); US5792616 (Fig 2); Cross-references: MIM:187395; NP_003203.1; NM_003212_1. 10 (14) CD21 (CR2 (Complement receptor 2) or C3DR (C3d/Epstein Barr virus receptor) or Hs.73792 Genbank accession no. M26004) Fujisaku et al. (1989) J. Biol. Chem.264 (4):2118- 2125); Weis J.J., et al. J. Exp. Med.167, 1047-1066, 1988; Moore M., et al. Proc. Natl. Acad. Sci. U.S.A.84, 9194-9198, 1987; Barel M., et al. Mol. Immunol.35, 1025-1031, 1998; Weis J.J., et al. Proc. Natl. Acad. Sci. U.S.A.83, 5639-5643, 1986; Sinha S.K., et al. (1993) J. 15 Immunol.150, 5311-5320; WO2004045520 (Example 4); US2004005538 (Example 1); WO2003062401 (Claim 9); WO2004045520 (Example 4); WO9102536 (Fig 9.1-9.9); WO2004020595 (Claim 1); Accession: P20023; Q13866; Q14212; EMBL; M26004; AAA35786.1. (15) CD79b (CD79B, CD79^, Igb (immunoglobulin-associated beta), B29, Genbank20 accession no. NM_000626 or 11038674) Proc. Natl. Acad. Sci. U.S.A. (2003) 100 (7):4126- 4131, Blood (2002) 100 (9):3068-3076, Muller et al. (1992) Eur. J. Immunol.22 (6):1621- 1625); WO2004016225 (claim 2, Fig 140); WO2003087768, US2004101874 (claim 1, page 102); WO2003062401 (claim 9); WO200278524 (Example 2); US2002150573 (claim 5, page 15); US5644033; WO2003048202 (claim 1, pages 306 and 309); WO 99/558658, US6534482 25 (claim 13, Fig 17A/B); WO200055351 (claim 11, pages 1145-1146); Cross-references: MIM:147245; NP_000617.1; NM_000626_1. (16) FcRH2 (IFGP4, IRTA4, SPAP1A (SH2 domain containing phosphatase anchor protein 1a), SPAP1B, SPAP1C, Genbank accession no. NM_030764, AY358130) Genome Res. 13 (10):2265-2270 (2003), Immunogenetics 54 (2):87-95 (2002), Blood 99 (8):2662-2669 30 (2002), Proc. Natl. Acad. Sci. U.S.A.98 (17):9772-9777 (2001), Xu, M.J., et al. (2001) Biochem. Biophys. Res. Commun.280 (3):768-775; WO2004016225 (Claim 2); WO2003077836; WO200138490 (Claim 5; Fig 18D-1-18D-2); WO2003097803 (Claim 12); WO2003089624 (Claim 25); Cross-references: MIM:606509; NP_110391.2; NM_030764_1. (17) HER2 (ErbB2, Genbank accession no. M11730) Coussens L., et al. Science (1985) 35 230(4730):1132-1139); Yamamoto T., et al. Nature 319, 230-234, 1986; Semba K., et al. Proc. 36 15077.006WO1 Natl. Acad. Sci. U.S.A.82, 6497-6501, 1985; Swiercz J.M., et al. J. Cell Biol.165, 869-880, 2004; Kuhns J.J., et al. J. Biol. Chem.274, 36422-36427, 1999; Cho H.-S., et al. Nature 421, 756-760, 2003; Ehsani A., et al. (1993) Genomics 15, 426-429; WO2004048938 (Example 2); WO2004027049 (Fig 1I); WO2004009622; WO2003081210; WO2003089904 (Claim 9); 5 WO2003016475 (Claim 1); US2003118592; WO2003008537 (Claim 1); WO2003055439 (Claim 29; Fig 1A-B); WO2003025228 (Claim 37; Fig 5C); WO200222636 (Example 13; Page 95-107); WO200212341 (Claim 68; Fig 7); WO200213847 (Page 71-74); WO200214503 (Page 114-117); WO200153463 (Claim 2; Page 41-46); WO200141787 (Page 15); WO200044899 (Claim 52; Fig 7); WO200020579 (Claim 3; Fig 2); US5869445 (Claim 3; Col 31-38); 10 WO9630514 (Claim 2; Page 56-61); EP1439393 (Claim 7); WO2004043361 (Claim 7); WO2004022709; WO200100244 (Example 3; Fig 4); Accession: P04626; EMBL; M11767; AAA35808.1. EMBL; M11761; AAA35808.1. (18) NCA (CEACAM6, Genbank accession no. M18728); Barnett T., et al. Genomics 3, 59-66, 1988; Tawaragi Y., et al. Biochem. Biophys. Res. Commun.150, 89-96, 1988; 15 Strausberg R.L., et al. Proc. Natl. Acad. Sci. U.S.A.99:16899-16903, 2002; WO2004063709; EP1439393 (Claim 7); WO2004044178 (Example 4); WO2004031238; WO2003042661 (Claim 12); WO200278524 (Example 2); WO200286443 (Claim 27; Page 427); WO200260317 (Claim 2); Accession: P40199; Q14920; EMBL; M29541; AAA59915.1. EMBL; M18728. (19) MDP (DPEP1, Genbank accession no. BC017023) Proc. Natl. Acad. Sci. U.S.A.99 20 (26):16899-16903 (2002)); WO2003016475 (Claim 1); WO200264798 (Claim 33; Page 85-87); JP05003790 (Fig 6-8); WO9946284 (Fig 9); Cross-references: MIM:179780; AAH17023.1; BC017023_1. (20) IL20R^ (IL20Ra, ZCYTOR7, Genbank accession no. AF184971); Clark H.F., et al. Genome Res.13, 2265-2270, 2003; Mungall A.J., et al. Nature 425, 805-811, 2003; Blumberg25 H., et al. Cell 104, 9-19, 2001; Dumoutier L., et al. J. Immunol.167, 3545-3549, 2001; Parrish- Novak J., et al. J. Biol. Chem.277, 47517-47523, 2002; Pletnev S., et al. (2003) Biochemistry 42:12617-12624; Sheikh F., et al. (2004) J. Immunol.172, 2006-2010; EP1394274 (Example 11); US2004005320 (Example 5); WO2003029262 (Page 74-75); WO2003002717 (Claim 2; Page 63); WO200222153 (Page 45-47); US2002042366 (Page 20-21); WO200146261 (Page 57- 30 59); WO200146232 (Page 63-65); WO9837193 (Claim 1; Page 55-59); Accession: Q9UHF4; Q6UWA9; Q96SH8; EMBL; AF184971; AAF01320.1. (21) Brevican (BCAN, BEHAB, Genbank accession no. AF229053) Gary S.C., et al. Gene 256, 139-147, 2000; Clark H.F., et al. Genome Res.13, 2265-2270, 2003; Strausberg R.L., et al. Proc. Natl. Acad. Sci. U.S.A.99, 16899-16903, 2002; US2003186372 (Claim 11); 35 US2003186373 (Claim 11); US2003119131 (Claim 1; Fig 52); US2003119122 (Claim 1; Fig 37 15077.006WO1 52); US2003119126 (Claim 1); US2003119121 (Claim 1; Fig 52); US2003119129 (Claim 1); US2003119130 (Claim 1); US2003119128 (Claim 1; Fig 52); US2003119125 (Claim 1); WO2003016475 (Claim 1); WO200202634 (Claim 1). (22) EphB2R (DRT, ERK, Hek5, EPHT3, Tyro5, Genbank accession no. NM_004442) 5 Chan, J. and Watt, V.M., Oncogene 6 (6), 1057-1061 (1991) Oncogene 10 (5):897-905 (1995), Annu. Rev. Neurosci.21:309-345 (1998), Int. Rev. Cytol.196:177-244 (2000)); WO2003042661 (Claim 12); WO200053216 (Claim 1; Page 41); WO2004065576 (Claim 1); WO2004020583 (Claim 9); WO2003004529 (Page 128-132); WO200053216 (Claim 1; Page 42); Cross-references: MIM:600997; NP_004433.2; NM_004442_1. 10 (23) ASLG659 (B7h, Genbank accession no. AX092328) US20040101899 (Claim 2); WO2003104399 (Claim 11); WO2004000221 (Fig 3); US2003165504 (Claim 1); US2003124140 (Example 2); US2003065143 (Fig 60); WO2002102235 (Claim 13; Page 299); US2003091580 (Example 2); WO200210187 (Claim 6; Fig 10); WO200194641 (Claim 12; Fig 7b); WO200202624 (Claim 13; Fig 1A-1B); US2002034749 (Claim 54; Page 45-46); 15 WO200206317 (Example 2; Page 320-321, Claim 34; Page 321-322); WO200271928 (Page 468-469); WO200202587 (Example 1; Fig 1); WO200140269 (Example 3; Pages 190-192); WO200036107 (Example 2; Page 205-207); WO2004053079 (Claim 12); WO2003004989 (Claim 1); WO200271928 (Page 233-234, 452-453); WO 0116318. (24) PSCA (Prostate stem cell antigen precursor, Genbank accession no. AJ297436) 20 Reiter R.E., et al. Proc. Natl. Acad. Sci. U.S.A.95, 1735-1740, 1998; Gu Z., et al. Oncogene 19, 1288-1296, 2000; Biochem. Biophys. Res. Commun. (2000) 275(3):783-788; WO2004022709; EP1394274 (Example 11); US2004018553 (Claim 17); WO2003008537 (Claim 1); WO200281646 (Claim 1; Page 164); WO 2003003906 (Claim 10; Page 288); WO 200140309 (Example 1; Fig 17); US 2001055751 (Example 1; Fig 1b); WO 200032752 (Claim 18; Fig 1); 25 WO 1998/51805 (Claim 17; Page 97); WO 1998/51824 (Claim 10; Page 94); WO 1998/40403 (Claim 2; Fig 1B); Accession: O43653; EMBL; AF043498; AAC39607.1. (25) GEDA (Genbank accession No. AY260763); AAP14954 lipoma HMGIC fusion- partner-like protein /pid=AAP14954.1 – Homo sapiens Species: Homo sapiens (human) WO2003054152 (Claim 20); WO2003000842 (Claim 1); WO2003023013 (Example 3, Claim 30 20); US2003194704 (Claim 45); Cross-references: GI:30102449; AAP14954.1; AY260763_1. (26) BAFF-R (B cell -activating factor receptor, BlyS receptor 3, BR3, Genbank accession No. AF116456); BAFF receptor /pid=NP_443177.1 – Homo sapiens Thompson, J.S., et al. Science 293 (5537), 2108-2111 (2001); WO2004058309; WO2004011611; WO2003045422 (Example; Page 32-33); WO2003014294 (Claim 35; Fig 6B); WO2003035846 35 (Claim 70; Page 615-616); WO200294852 (Col 136-137); WO200238766 (Claim 3; Page 133); 38 15077.006WO1 WO200224909 (Example 3; Fig 3); Cross-references: MIM:606269; NP_443177.1; NM_052945_1; AF132600. (27) CD22 (B-cell receptor CD22-B isoform, BL-CAM, Lyb-8, Lyb8, SIGLEC-2, FLJ22814, Genbank accession No. AK026467); Wilson et al. (1991) J. Exp. Med.173:137-146; 5 WO2003072036 (Claim 1; Fig 1); Cross-references: MIM:107266; NP_001762.1; NM_001771_1. (28) CD79a (CD79A, CD79^, immunoglobulin-associated alpha, a B cell-specific protein that covalently interacts with Ig beta (CD79B) and forms a complex on the surface with Ig M molecules, transduces a signal involved in B-cell differentiation), pI: 4.84, MW: 25028 10 TM: 2 [P] Gene Chromosome: 19q13.2, Genbank accession No. NP_001774.10) WO2003088808, US20030228319; WO2003062401 (claim 9); US2002150573 (claim 4, pages 13-14); WO9958658 (claim 13, Fig 16); WO9207574 (Fig 1); US5644033; Ha et al. (1992) J. Immunol.148(5):1526-1531; Mueller et al. (1992) Eur. J. Biochem.22:1621-1625; Hashimoto et al. (1994) Immunogenetics 40(4):287-295; Preud’homme et al. (1992) Clin. Exp. Immunol. 15 90(1):141-146; Yu et al. (1992) J. Immunol.148(2) 633-637; Sakaguchi et al. (1988) EMBO J. 7(11):3457-3464. (29) CXCR5 (Burkitt’s lymphoma receptor 1, a G protein-coupled receptor that is activated by the CXCL13 chemokine, functions in lymphocyte migration and humoral defense, plays a role in HIV-2 infection and perhaps development of AIDS, lymphoma, myeloma, and 20 leukemia); 372 aa, pI: 8.54 MW: 41959 TM: 7 [P] Gene Chromosome: 11q23.3, Genbank accession No. NP_001707.1) WO 2004040000; WO2004/015426; US2003105292 (Example 2); US6555339 (Example 2); WO 2002/61087 (Fig 1); WO200157188 (Claim 20, page 269); WO200172830 (pages 12-13); WO 2000/22129 (Example 1, pages 152-153, Example 2, pages 254-256); WO 199928468 (claim 1, page 38); US 5440021 (Example 2, col 49-52); 25 WO9428931 (pages 56-58); WO 1992/17497 (claim 7, Fig 5); Dobner et al. (1992) Eur. J. Immunol.22:2795-2799; Barella et al. (1995) Biochem. J.309:773-779. (30) HLA-DOB (Beta subunit of MHC class II molecule (Ia antigen) that binds peptides and presents them to CD4+ T lymphocytes); 273 aa, pI: 6.56 MW: 30820 TM: 1 [P] Gene Chromosome: 6p21.3, Genbank accession No. NP_002111.1) Tonnelle et al. (1985) EMBO J. 30 4(11):2839-2847; Jonsson et al. (1989) Immunogenetics 29(6):411-413; Beck et al. (1992) J. Mol. Biol.228:433-441; Strausberg et al. (2002) Proc. Natl. Acad. Sci USA 99:16899-16903; Servenius et al. (1987) J. Biol. Chem.262:8759-8766; Beck et al. (1996) J. Mol. Biol.255:1-13; Naruse et al. (2002) Tissue Antigens 59:512-519; WO9958658 (claim 13, Fig 15); US6153408 (Col 35-38); US5976551 (col 168-170); US6011146 (col 145-146); Kasahara et al. (1989) 35 Immunogenetics 30(1):66-68; Larhammar et al. (1985) J. Biol. Chem.260(26):14111-14119. 39 15077.006WO1 (31) P2X5 (Purinergic receptor P2X ligand-gated ion channel 5, an ion channel gated by extracellular ATP, may be involved in synaptic transmission and neurogenesis, deficiency may contribute to the pathophysiology of idiopathic detrusor instability); 422 aa), pI: 7.63, MW: 47206 TM: 1 [P] Gene Chromosome: 17p13.3, Genbank accession No. NP_002552.2) Le et al. 5 (1997) FEBS Lett.418(1-2):195-199; WO2004047749; WO2003072035 (claim 10); Touchman et al. (2000) Genome Res.10:165-173; WO200222660 (claim 20); WO2003093444 (claim 1); WO2003087768 (claim 1); WO2003029277 (page 82). (32) CD72 (B-cell differentiation antigen CD72, Lyb-2), pI: 8.66, MW: 40225 TM: 1 [P] Gene Chromosome: 9p13.3, Genbank accession No. NP_001773.1) WO2004042346 (claim 10 65); WO 2003/026493 (pages 51-52, 57-58); WO 2000/75655 (pages 105-106); Von Hoegen et al. (1990) J. Immunol.144(12):4870-4877; Strausberg et al. (2002) Proc. Natl. Acad. Sci USA 99:16899-16903. (33) LY64 (Lymphocyte antigen 64 (RP105), type I membrane protein of the leucine rich repeat (LRR) family, regulates B-cell activation and apoptosis, loss of function is associated 15 with increased disease activity in patients with systemic lupus erythematosus); 661 aa, pI: 6.20, MW: 74147 TM: 1 [P] Gene Chromosome: 5q12, Genbank accession No. NP_005573.1) US2002193567; WO9707198 (claim 11, pages 39-42); Miura et al. (1996) Genomics 38(3):299- 304; Miura et al. (1998) Blood 92:2815-2822; WO2003083047; WO9744452 (claim 8, pages 57-61); WO200012130 (pages 24-26). 20 (34) FcRH1 (Fc receptor-like protein 1, a putative receptor for the immunoglobulin Fc domain that contains C2 type Ig-like and ITAM domains, may have a role in B-lymphocyte differentiation); 429 aa, pI: 5.28, MW: 46925 TM: 1 [P] Gene Chromosome: 1q21-1q22, Genbank accession No. NP_443170.1) WO2003077836; WO200138490 (claim 6, Fig 18E-1- 18-E-2); Davis et al. (2001) Proc. Natl. Acad. Sci USA 98(17):9772-9777; WO2003089624 25 (claim 8); EP1347046 (claim 1); WO2003089624 (claim 7). (35) IRTA2 (Immunoglobulin superfamily receptor translocation associated 2, a putative immunoreceptor with possible roles in B cell development and lymphomagenesis; deregulation of the gene by translocation occurs in some B cell malignancies); 977 aa, pI: 6.88 MW: 106468 TM: 1 [P] Gene Chromosome: 1q21, Genbank accession No. Human:AF343662, AF343663, 30 AF343664, AF343665, AF369794, AF397453, AK090423, AK090475, AL834187, AY358085; Mouse:AK089756, AY158090, AY506558; NP_112571.1. WO2003024392 (claim 2, Fig 97); Nakayama et al. (2000) Biochem. Biophys. Res. Commun.277(1):124-127; WO2003077836; WO200138490 (claim 3, Fig 18B-1-18B-2). (36) TENB2 (TMEFF2, tomoregulin, TPEF, HPP1, TR, putative transmembrane 35 proteoglycan, related to the EGF/heregulin family of growth factors and follistatin); 374 aa, 40 15077.006WO1 NCBI Accession: AAD55776, AAF91397, AAG49451, NCBI RefSeq: NP_057276; NCBI Gene: 23671; OMIM: 605734; SwissProt Q9UIK5; Genbank accession No. AF179274; AY358907, CAF85723, CQ782436 WO 2004074320; JP 2004113151; WO 2003042661; WO2003009814; EP1295944 (pages 69-70); WO 200230268 (page 329); WO 200190304; 5 US2004249130; US 2004022727; WO 2004063355; US 2004197325; US2003232350; US2004005563; US 2003124579; Horie et al. (2000) Genomics 67:146-152; Uchida et al. (1999) Biochem. Biophys. Res. Commun.266:593-602; Liang et al. (2000) Cancer Res.60:4907- 12; Glynne-Jones et al. (2001) Int J Cancer. Oct 15;94(2):178-84. (37) PMEL17 (silver homolog; SILV; D12S53E; PMEL17; SI; SIL); ME20; gp100) 10 BC001414; BT007202; M32295; M77348; NM_006928; McGlinchey, R.P. et al. (2009) Proc. Natl. Acad. Sci. U.S.A.106 (33), 13731-13736; Kummer, M.P. et al. (2009) J. Biol. Chem.284 (4), 2296-2306. (38) TMEFF1 (transmembrane protein with EGF-like and two follistatin-like domains 1; Tomoregulin-1); H7365; C9orf2; C9ORF2; U19878; X83961; NM_080655; NM_003692; 15 Harms, P.W. (2003) Genes Dev.17 (21), 2624-2629; Gery, S. et al. (2003) Oncogene 22 (18):2723-2727. (39) GDNF-Ra1 (GDNF family receptor alpha 1; GFRA1; GDNFR; GDNFRA; RETL1; TRNR1; RET1L; GDNFR-alpha1; GFR-ALPHA-1); U95847; BC014962; NM_145793 NM_005264; Kim, M.H. et al. (2009) Mol. Cell. Biol.29 (8), 2264-2277; Treanor, J.J. et al. 20 (1996) Nature 382 (6586):80-83. (40) Ly6E (lymphocyte antigen 6 complex, locus E; Ly67,RIG-E,SCA-2,TSA-1); NP_002337.1; NM_002346.2; de Nooij-van Dalen, A.G. et al. (2003) Int. J. Cancer 103 (6), 768-774; Zammit, D.J. et al. (2002) Mol. Cell. Biol.22 (3):946-952. (41) TMEM46 (shisa homolog 2 (Xenopus laevis); SHISA2); NP_001007539.1; 25 NM_001007538.1; Furushima, K. et al. (2007) Dev. Biol.306 (2), 480-492; Clark, H.F. et al. (2003) Genome Res.13 (10):2265-2270. (42) Ly6G6D (lymphocyte antigen 6 complex, locus G6D; Ly6-D, MEGT1); NP_067079.2; NM_021246.2; Mallya, M. et al. (2002) Genomics 80 (1):113-123; Ribas, G. et al. (1999) J. Immunol.163 (1):278-287. 30 (43) LGR5 (leucine-rich repeat-containing G protein-coupled receptor 5; GPR49, GPR67); NP_003658.1; NM_003667.2; Salanti, G. et al. (2009) Am. J. Epidemiol.170 (5):537- 545; Yamamoto, Y. et al. (2003) Hepatology 37 (3):528-533. (44) RET (ret proto-oncogene; MEN2A; HSCR1; MEN2B; MTC1; PTC; CDHF12; Hs.168114; RET51; RET-ELE1); NP_066124.1; NM_020975.4; Tsukamoto, H. et al. (2009) 35 Cancer Sci.100 (10):1895-1901; Narita, N. et al. (2009) Oncogene 28 (34):3058-3068. 41 15077.006WO1 (45) LY6K (lymphocyte antigen 6 complex, locus K; LY6K; HSJ001348; FLJ35226); NP_059997.3; NM_017527.3; Ishikawa, N. et al. (2007) Cancer Res.67 (24):11601-11611; de Nooij-van Dalen, A.G. et al. (2003) Int. J. Cancer 103 (6):768-774. (46) GPR19 (G protein-coupled receptor 19; Mm.4787); NP_006134.1; NM_006143.2; 5 Montpetit, A. and Sinnett, D. (1999) Hum. Genet.105 (1-2):162-164; O’Dowd, B.F. et al. (1996) FEBS Lett.394 (3):325-329. (47) GPR54 (KISS1 receptor; KISS1R; GPR54; HOT7T175; AXOR12); NP_115940.2; NM_032551.4; Navenot, J.M. et al. (2009) Mol. Pharmacol.75 (6):1300-1306; Hata, K. et al. (2009) Anticancer Res.29 (2):617-623. 10 (48) ASPHD1 (aspartate beta-hydroxylase domain containing 1; LOC253982); NP_859069.2; NM_181718.3; Gerhard, D.S. et al. (2004) Genome Res.14 (10B):2121-2127. (49) Tyrosinase (TYR; OCAIA; OCA1A; tyrosinase; SHEP3); NP_000363.1; NM_000372.4; Bishop, D.T. et al. (2009) Nat. Genet.41 (8):920-925; Nan, H. et al. (2009) Int. J. Cancer 125 (4):909-917. 15 (50) TMEM118 (ring finger protein, transmembrane 2; RNFT2; FLJ14627); NP_001103373.1; NM_001109903.1; Clark, H.F. et al. (2003) Genome Res.13 (10):2265-2270; Scherer, S.E. et al. (2006) Nature 440 (7082):346-351. (51) GPR172A (G protein-coupled receptor 172A; GPCR41; FLJ11856; D15Ertd747e); NP_078807.1; NM_024531.3; Ericsson, T.A. et al. (2003) Proc. Natl. Acad. Sci. U.S.A.100 20 (11):6759-6764; Takeda, S. et al. (2002) FEBS Lett.520 (1-3):97-101. (52) CD33, a member of the sialic acid binding, immunoglobulin-like lectin family, is a 67-kDa glycosylated transmembrane protein. CD33is expressed on most myeloid and monocytic leukemia cells in addition to committed myelomonocytic and erythroid progenitor cells. It is not seen on the earliest pluripotent stem cells, mature granulocytes, lymphoid cells, or 25 nonhematopoietic cells (Sabbath et al., (1985) J. Clin. Invest.75:756-56; Andrews et al., (1986) Blood 68:1030-5). CD33 contains two tyrosine residues on its cytoplasmic tail, each of which is followed by hydrophobic residues similar to the immunoreceptor tyrosine-based inhibitory motif (ITIM) seen in many inhibitory receptors. (53) CLL-1 (CLEC12A, MICL, and DCAL2), encodes a member of the C-type lectin/C- 30 type lectin-like domain (CTL/CTLD) superfamily. Members of this family share a common protein fold and have diverse functions, such as cell adhesion, cell-cell signaling, glycoprotein turnover, and roles in inflammation and immune response. The protein encoded by this gene is a negative regulator of granulocyte and monocyte function. Several alternatively spliced transcript variants of this gene have been described, but the full-length nature of some of these variants has 35 not been determined. This gene is closely linked to other CTL/CTLD superfamily members in 42 15077.006WO1 the natural killer gene complex region on chromosome 12p13 (Drickamer K (1999) Curr. Opin. Struct. Biol.9 (5):585–90; van Rhenen A, et al., (2007) Blood 110 (7):2659–66; Chen CH, et al. (2006) Blood 107 (4):1459–67; Marshall AS, et al. (2006) Eur. J. Immunol.36 (8):2159–69; Bakker AB, et al. (2005) Cancer Res.64 (22):8443–50; Marshall AS, , et al. (2004) J. Biol. 5 Chem.279 (15):14792–802). CLL-1 has been shown to be a type II transmembrane receptor comprising a single C-type lectin-like domain (which is not predicted to bind either calcium or sugar), a stalk region, a transmembrane domain and a short cytoplasmic tail containing an ITIM motif. (54) TROP2 (tumor-associated calcium signal transducer 2) is a transmembrane 10 glycoprotein encoded by the TACSTD2 gene (Linnenbach AJ, et al (1993) Mol Cell Biol.13(3): 1507–15; Calabrese G, et al (2001) Cytogenet Cell Genet.92(1–2): 164–5). TROP2 is an intracellular calcium signal transducer that is differentially expressed in many cancers. It signals cells for self-renewal, proliferation, invasion, and survival. It has stem cell-like qualities. TROP2 is expressed in many normal tissues, though in contrast, it is overexpressed in many cancers 15 (Ohmachi T, et al., (2006) Clin. Cancer Res., 12(10):3057-3063; Muhlmann G, et al., (2009) J. Clin. Pathol., 62(2):152-158; Fong D, et al., (2008) Br. J. Cancer, 99(8):1290-1295; Fong D, et al., (2008) Mod. Pathol., 21(2):186-191; Ning S, et al., (2013) Neurol. Sci., 34(10):1745-1750). Overexpression of TROP2 is of prognostic significance. Several ligands have been proposed that interact with TROP2. TROP2 signals the cells via different pathways and it is transcriptionally 20 regulated by a complex network of several transcription factors. Human TROP2 (TACSTD2: tumor-associated calcium signal transducer 2, GA733-1, EGP-1, M1S1; hereinafter, referred to as hTROP2) is a single-pass transmembrane type 1 cell membrane protein consisting of 323 amino acid residues. While the presence of a cell membrane protein involved in immune resistance, which is common to human trophoblasts and cancer cells 25 (Faulk W P, et al. (1978), Proc. Natl. Acad. Sci.75(4):1947-1951), has previously been suggested, an antigen molecule recognized by a monoclonal antibody against a cell membrane protein in a human choriocarcinoma cell line was identified and designated as TROP2 as one of the molecules expressed In human trophoblasts (Lipinski M, et al. (1981), Proc. Natl. Acad. Sci. 78(8):5147-5150). This molecule was also designated as tumor antigen GA733-1 recognized by 30 a mouse monoclonal antibody GA733 (Linnenbach A J, et al., (1989) Proc. Natl. Acad. Sci. 86(1):27-31) obtained by immunization with a gastric cancer cell line or an epithelial glycoprotein (EGP-1; Basu A, et al., Int. J. Cancer, 62 (4), 472-479 (1995)) recognized by a mouse monoclonal antibody RS7-3G11 obtained by immunization with non-small cell lung cancer cells. In 1995, however, the TROP2 gene was cloned, and all of these molecules were 35 confirmed to be identical molecules (Fornaro M, et al., (1995) Int. J. Cancer, 62(5):610-618). 43 15077.006WO1 The DNA sequence and amino acid sequence of hTROP2 are available on a public database and can be referred to, for example, under Accession Nos. NM_002353 and NP_002344 (NCBI). TPI-VHL COMPOUNDS The antibody conjugate composition of the invention comprises a target protein binder 5 and VHL ligand moiety (TPI-VHL). The target protein of interest (TPI) binder is covalently attached to the VHL ligand by a spacer unit. Exemplary target proteins include RIPK2, Bcl-XL, KRAS, and WEE1. Receptor-interacting protein kinase 2 (RIPK2) is a serine/threonine kinase integral to innate immune signaling via the NOD1/2 pathways, with emerging roles in tumor biology. 10 RIPK2 activation promotes chronic inflammation through NF-κB and MAPK signaling, fostering tumor progression by enhancing immune cell infiltration, RNA methylation, and tumor stemness. Pro-survival protein Bcl-xL (B-cell lymphoma-extra large), is a key regulator of the intrinsic pathway and promotes cell survival by inhibiting pro-apoptotic counterparts. 15 KRAS (Kirsten rat sarcoma virus) is a proto-oncogene that encodes the K‑Ras protein, a small GTPase that is a critical component of the RAS/MAPK signaling cascade. This protein normally functions as a molecular switch cycling between an inactive GDP‑bound state and an active GTP‑bound state. WEE1, a tyrosine kinase regulating the G2/M checkpoint, ensures genomic stability by 20 phosphorylating CDK1/2. Overexpression in endometrial cancer and melanoma confers poor prognosis by enabling DNA repair evasion and fostering replication stress. The antibody conjugate composition of the present disclosure may be prepared from a compound comprising a target protein binder (TPI) covalently attached to a VHL ligand by a spacer unit (Sp) having Formula II: 25 TPI−Sp−VHL II or a pharmaceutically acceptable salt, stereoisomer, mixture of stereoisomers, tautomer, or isotopic analog thereof, wherein: TPI binds to RIPK2, KRAS, Bcl-XL, or WEE1; VHL ligand has Formula IIa: 44 15077.006WO1 IIa wherein A is a cyclic structure selected from C₃−C₂₀ carbocyclyl, C₆−C₂₀ aryl, C₁−C₂₀ heteroaryl, and C₂-C₂₀ heterocyclyl, each of which are substituted with one or more groups independently selected from H, F, Cl, Br, I, −CN, −NO₂, C₁−C₁₂ alkyl, C₂−C₁₂ alkenyl, C₂−C₁₂ 5 alkynyl, C₁−C₁₂ heteroalkyl, −(C₁−C₁₂ heteroalkyldiyl)−(C₆−C₂₀ aryl), −(C₁−C₁₂ heteroalkyldiyl)−(C₆−C₂₀ aryldiyl)−(C₁−C₁₂ heteroalkyl), −(C₁−C₁₂ heteroalkyldiyl)−(C₆−C₂₀ aryldiyl)−(C₂-C₂₀ heterocyclyl), (C₁−C₆ alkyldiyl)−(C₆−C₂₀ aryl), −(C₁−C₆ alkyldiyl)−NR^aR^b, −(C₁−C₆ alkyldiyl)−OR^a, (C₁−C₆ alkyldiyl)−(C₃−C₂₀ carbocyclyl), (C₁-C₆ alkyldiyl)−(C₂-C₂₀ heterocyclyl), (C₁−C₆ alkyldiyl)−(C₁−C₂₀ heteroaryl), C₆−C₂₀ aryl, C₃−C₂₀ carbocyclyl, C₂−C₂₀ 10 heterocyclyl, C₁−C₂₀ heteroaryl, −C(=NH)NH(OH), −C(=NH)NH₂, −C(=O)NR^aR^b, −C(=O)NR^a−NR^aR^b, −C(=O)NH(C₁-C₆ alkyldiyl)−NR^aR^b, −C(=O)OR^a, −NR^aR^b, −NR^aS(O)₂R^a, −NO2, −OR^a, −OC(=O)R^a, −SR^a, −S(O)R^a, −S(O)2R^a, −S(O)2NR^a, and −S(O)3H; R^a is independently selected from H, C1−C6 alkyl, phenyl, and benzyl, wherein phenyl and benzyl are optionally substituted with one or more groups independently selected from the 15 group consisting of F, Cl, −CN, C₁−C₁₂ alkyl, C₂−C₁₂ alkenyl, and C₂−C₁₂ alkynyl; or where two R^a groups form a five-, six-, or seven-membered heteroaryl or heterocyclyl ring; R^b is independently selected from H, OH, C1−C6 alkyl, phenyl, and benzyl, wherein phenyl and benzyl are optionally substituted with one or more groups independently selected 20 from the group consisting of F, Cl, −CN, C1−C12 alkyl, C2−C12 alkenyl, and C2−C12 alkynyl; R^c is selected from OH and −SO2F; R^d is selected from H and F; Cyc is a ring structure selected from the group consisting of C₃−C₂₀ carbocyclyl, C₆−C₂₀ aryl, C₂-C₂₀ heterocyclyl, C₁−C₂₀ heteroaryl, and −(C₁−C₂₀ heteroaryldiyl)−(C₁−C₁₂ 25 heteroalkyl); n is 0 or 1; 45 15077.006WO1 R¹ is selected from the group consisting of H, C1−C12 alkyl, and C1−C12 heteroalkyl; R² is selected from the group consisting of H, C1−C12 alkyl, and C1−C12 heteroalkyl; or where R² forms a five- to ten-membered aryl, carbocyclyl, heteroaryl or heterocyclyl ring with A; 5 X¹ is selected from the group consisting of H, −NHC(=O)−, (C₃−C₂₀ carbocyclyl)− C(=O)NH−, C₁−C₁₂ heteroalkyl, and C₁−C₂₀ heteroaryl; or where X¹ forms a five-, six-, or seven-membered heteroaryl or heterocyclyl ring with R¹; Sp is selected from the group consisting of a bond, O, NH, C₁−C₁₂ alkyldiyl, C₁−C₆₀ heteroalkyldiyl, C₃−C₂₀ carbocyclyldiyl, C₂-C₂₀ heterocyclyldiyl, C₆-C₂₀ aryldiyl, C₁−C₄₀ 10 heteroaryldiyl, −(C₃−C₂₀ carbocyclyldiyl)−(C₁−C₆₀ heteroalkyldiyl)−, −(C₃−C₂₀ carbocyclyldiyl)−(C₁−C₁₂ alkyldiyl)−, −(C₂-C₂₀ heterocyclyldiyl)−(C₁−C₆₀ heteroalkyldiyl)−, −(C₂-C₂₀ heterocyclyldiyl)−(C₁−C₁₂ alkyldiyl)−, −(C₂-C₂₀ heterocyclyldiyl)−(C₂-C₂₀ heterocyclyldiyl)−, −(C₁−C₂₀ heteroaryldiyl)−(C₃−C₂₀ carbocyclyldiyl)−(C₁−C₆₀ heteroalkyldiyl)−, a solubilizing unit, and combinations thereof, where the solubilizing unit is 15 selected from polyglycine, polysarcosine, polyethyleneoxy (PEG), a glycoside, C₁−C₆₀ heteroalkyldiyl, and combinations thereof; one of A, R^a, R^b, Cyc, R¹, R², and X¹ is attached to Sp; and each alkyl, alkyldiyl, alkenyl, alkenyldiyl, alkynyl, alkynyldiyl, heteroalkyl, heteroalkyldiyl, aryl, aryldiyl, carbocyclyl, carbocyclyldiyl, heterocyclyl, heterocyclyldiyl, 20 heteroaryl, and heteroaryldiyl is independently and optionally substituted with one or more groups selected from F, Cl, Br, I, −CN, −CH3, −CH2CH3, −CH=CH2, −C^CH, −C^CCH3, − C^CCH₂NHCH₃, −CH₂CH₂CH₃, −CH(CH₃)₂, −CH₂CH(CH₃)₂, −CH₂OH, −CH₂OCH₃, − CH₂CH₂OH, −C(CH₃)₂OH, −CH(OH)CH(CH₃)₂, −C(CH₃)₂CH₂OH, −CH₂CH₂SO₂CH₃, − CH₂OP(O)(OH)₂, −CH₂F, −CHF₂, −CF₃, −CH₂CF₃, −CH₂CHF₂, −CH(CH₃)CN, −C(CH₃)₂CN, −25 CH₂CN, −CH₂NH₂, −CH₂NHSO₂CH₃, −CH₂NHCH₃, −CH₂N(CH₃)₂, −CO₂H, −COCH₃, − CO₂CH₃, −CO₂C(CH₃)₃, −COCH(OH)CH₃, −CONH₂, −CONHCH₃, −CON(CH₃)₂, − C(CH₃)₂CONH₂, −NH₂, −NHCH₃, −N(CH₃)₂, −NHCOCH₃, −N(CH₃)COCH₃, −NHS(O)₂CH₃, − N(CH₃)C(CH₃)₂CONH₂, −N(CH₃)CH₂CH₂S(O)₂CH₃, −NHC(=NH)H, −NHC(=NH)CH₃, − NHC(=NH)NH₂, −NHC(=O)NH₂, −NO₂, =O, −OH, −OCH₃, −OCH₂CH₃, −OCH₂CH₂OCH₃, − 30 OCH2CH2OH, −OCH2CH2N(CH3)2, −OCH2F, −OCHF2, −OCF3, −OP(O)(OH)2, −S(O)2N(CH3)2, −SCH3, −S(O)2CH3, and −S(O)3H. An exemplary embodiment of the Formula II compound includes wherein the VHL ligand has Formula IIb: 46 15077.006WO1 . An exemplary embodiment of the Formula II compound includes wherein the VHL ligand has Formula IIc: 5 wherein: R³, R⁴, R⁵, and R⁶ are independently selected from the group consisting of H, F, Cl, Br, I, −CN, C1−C12 alkyl, C2−C12 alkenyl, C2−C12 alkynyl, C1−C12 heteroalkyl, −(C1−C12 heteroalkyldiyl)−(C6−C20 aryl), −(C1−C12 heteroalkyldiyl)−(C6−C20 aryldiyl)−(C1−C12 heteroalkyl), −(C1−C12 heteroalkyldiyl)−(C6−C20 aryldiyl)−(C2-C20 heterocyclyl), (C1−C6 10 alkyldiyl)−(C6−C20 aryl), −(C1−C6 alkyldiyl)−NR^aR^b, −(C1−C6 alkyldiyl)−OR^a, (C1−C6 alkyldiyl)−(C3−C20 carbocyclyl), (C1-C6 alkyldiyl)−(C2-C20 heterocyclyl), (C1−C6 alkyldiyl)−(C1−C20 heteroaryl), C6−C20 aryl, C3−C20 carbocyclyl, C2−C20 heterocyclyl, C1−C20 heteroaryl, −C(=NH)NH(OH), −C(=NH)NH2, −C(=O)NR^aR^b, −C(=O)NR^a−NR^aR^b, −C(=O)NH(C1-C6 alkyldiyl)−NR^aR^b, −C(=O)OR^a, −NR^aR^b, −NO2, −OR^a, −OC(=O)R^a, −SR^a, 15 −S(O)R^a, −S(O)2R^a, −S(O)2NR^a, −S(O)3H, and Sp; R^a is independently selected from H, C₁−C₆ alkyl, phenyl, and benzyl, wherein phenyl and benzyl are optionally substituted with one or more groups independently selected from the group consisting of F, Cl, −CN, C1−C12 alkyl, C2−C12 alkenyl, C2−C12 alkynyl, and Sp; or where two R^a groups form a five-, six-, or seven-membered heteroaryl or heterocyclyl 20 group; 47 15077.006WO1 R^b is independently selected from H, OH, C1−C6 alkyl, phenyl, and benzyl, wherein phenyl and benzyl are optionally substituted with one or more groups independently selected from the group consisting of F, Cl, −CN, C₁−C₁₂ alkyl, C₂−C₁₂ alkenyl, C₂−C₁₂ alkynyl, and Sp; or where R³ and R⁴ together form a five-membered or six-membered heteroaryl or 5 heterocyclyl group comprising one or more heteroatoms independently selected from N, O, P and S; X¹ is selected from the group consisting of H, and −NHCO−; or where X¹ forms a five-, six-, or seven-membered heteroaryl or heterocyclyl ring with R¹; one of X¹, R¹, R², R³, R⁴, R⁵, R⁶, R^a, R^b, and Cyc is attached to Sp; and 10 each alkyl, alkyldiyl, alkenyl, alkenyldiyl, alkynyl, alkynyldiyl, heteroalkyl, heteroalkyldiyl, aryl, aryldiyl, carbocyclyl, carbocyclyldiyl, heterocyclyl, heterocyclyldiyl, heteroaryl, and heteroaryldiyl is independently and optionally substituted with one or more groups selected from F, Cl, Br, I, −CN, −CH₃, −CH₂CH₃, −CH=CH₂, −C^CH, −C^CCH₃, − CH₂CH₂CH₃, −CH(CH₃)₂, −CH₂CH(CH₃)₂, −CH₂OH, −CH₂OCH₃, −CH₂CH₂OH, −C(CH₃)₂OH, 15 −CH(OH)CH(CH₃)₂, −C(CH₃)₂CH₂OH, −CH₂CH₂SO₂CH₃, −CH₂OP(O)(OH)₂, −CH₂F, −CHF₂, −CF₃, −CH₂CF₃, −CH₂CHF₂, −CH(CH₃)CN, −C(CH₃)₂CN, −CH₂CN, −CH₂NH₂, − CH₂NHSO₂CH₃, −CH₂NHCH₃, −CH₂N(CH₃)₂, −CO₂H, −COCH₃, −CO₂CH₃, −CO₂C(CH₃)₃, − COCH(OH)CH₃, −CONH₂, −CONHCH₃, −CON(CH₃)₂, −CONHS(O)₂N(CH₃)₂, − C(CH3)2CONH2, −NH2, −NHCH3, −N(CH3)2, −NHCOCH3, −N(CH3)COCH3, −NHS(O)2CH3, −20 N(CH3)C(CH3)2CONH2, −N(CH3)CH2CH2S(O)2CH3, −NHC(=NH)H, −NHC(=NH)CH3, − NHC(=NH)NH2, −NHC(=O)NH2, −NO2, =O, −OH, −OCH3, −OCH2CH3, −OCH2CH2OCH3, − OCH2CH2OH, −OCH2CH2N(CH3)2, −OCH2F, −OCHF2, −OCF3, −OP(O)(OH)2, −S(O)2N(CH3)2, −SCH3, −S(O)2CH3, and −S(O)3H. Exemplary target protein binder and VHL ligand (TPI-VHL) compounds of Tables 1a-d 25 were prepared and characterized according to the Examples herein. Each compound in Tables 1a-d were characterized by mass spectrometry and demonstrated to have the correct parent ion and mass. Certain exemplary TPI-VHL compounds of Tables 1a-d were tested for their effects in inhibiting cellular proliferation, including CAL51, a human breast cancer cell line. Target protein binder and VHL ligand (TPI-VHL) compounds may be converted to target protein binder 30 and VHL ligand linker ((TPI-VHL)-L) compounds for conjugation with an antibody to form Degrader Antibody Conjugate (DAC) compositions. Table 1a RIPK2-VHL compounds (RV) 48 15077.006WO1 Table 1b1 Bcl-xL-VHL compounds (BXV) 49 15077.006WO1 50 15077.006WO1 51 15077.006WO1 52 15077.006WO1 53 15077.006WO1 54 15077.006WO1 55 15077.006WO1 56 15077.006WO1 57 15077.006WO1 58 15077.006WO1 59 15077.006WO1 60 15077.006WO1 61 15077.006WO1 62 15077.006WO1 63 15077.006WO1 Table 1b2 Bcl-xL-VHL compounds (BXV) 64 15077.006WO1 65 15077.006WO1 66 15077.006WO1 67 15077.006WO1 68 15077.006WO1 69 15077.006WO1 70 15077.006WO1 71 15077.006WO1 72 15077.006WO1 73 15077.006WO1 74 15077.006WO1 75 15077.006WO1 76 15077.006WO1 Table 1c KRAS-VHL compounds (KV) 77 15077.006WO1 78 15077.006WO1 79 15077.006WO1 80 15077.006WO1 81 15077.006WO1 82 15077.006WO1 83 15077.006WO1 84 15077.006WO1 85 15077.006WO1 86 15077.006WO1 87 15077.006WO1 88 15077.006WO1 89 15077.006WO1 90 15077.006WO1 91 15077.006WO1 92 15077.006WO1 93 15077.006WO1 94 15077.006WO1 95 15077.006WO1 96 15077.006WO1 97 15077.006WO1 Table 1d WEE1-VHL compounds (WV) 98 15077.006WO1 99 15077.006WO1 100 15077.006WO1 101 15077.006WO1 102 15077.006WO1 103 15077.006WO1 104 15077.006WO1 105 15077.006WO1 106 15077.006WO1 107 15077.006WO1 ANTIBODY LINKER UNITS The antibody conjugate composition of the invention comprises an antibody linker covalently attaching the target protein binder and VHL ligand (TPI-VHL) moiety to an antibody. 5 The TPI-VHL moiety is attached to a linker unit to prepare target protein binder and VHL ligand linker ((TPI-VHL)-L) compounds for conjugation with an antibody (Ab) to form Degrader Antibody Conjugate (DAC) compositions. In one aspect, the antibody linker of the antibody conjugates and target protein binder and VHL ligand linker compound comprises a branched phenyl maleimide group and other 10 linker units described in WO 2023/173121, which is incorporated by reference herein. In some embodiments, an immolative group (IM), and TPI-VHL moiety together have the following structure: 108 15077.006WO1 5 wherein indicates an attachment site to the remainder of the molecule (i.e., a compound of Structure (I) or conjugate of Structure (II)) and a TPI-VHL moiety. Amino acids of embodiments above may be replaced or used in addition to other amino acids, in some 10 embodiments, a peptide group (PEP) 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 an immolative group is PABC. 109 15077.006WO1 In some embodiments, the phenyl portion of the PABC is substituted with one or more substituents. In some embodiments, the substituents have one of the following structures: , , , 5 In some embodiments, an immolative group comprises one of the following structures: . In some embodiments, a trigger element, an immolative unit (IM), and TPI-VHL moiety together comprise one of the following structures: 10 , . In some embodiments, an immolative unit (IM) has a structure selected from the following: 110 15077.006WO1 . 5 The structures above show a substitution pattern of 1, 3, 4 on the phenyl ring of an immolative unit (IM). In some embodiments, a substitution pattern may be 1, 2, 4 (i.e., 1 being a linkage to a TPI-VHL, 2 being a linkage to the remainder of the molecule and 4 being a linkage 111 15077.006WO1 to the carbohydrate) or 1, 3, 5 (i.e., 1 being a linkage to a TPI-VHL, 3 being a linkage to the remainder of the molecule and 4 being a linkage to the carbohydrate). Although cleavable linkers (e.g., linkers with trigger elements or immolative unit) can provide certain advantages, linkers need not be cleavable. For non-cleavable linkers, a TPI-VHL 5 release may not depend on the differential properties between the plasma and some cytoplasmic compartments. The release of a TPI-VHL can occur after internalization of the conjugate of Structure (II) via antigen-mediated endocytosis and delivery to lysosomal compartment, where the targeting moiety (or binding fragment thereof) can be degraded to the level of amino acids through intracellular proteolytic degradation. This process can release a TPI-VHL moiety or 10 TPI-VHL moiety derivative. A TPI-VHL moiety or TPI-VHL moiety derivative can be more hydrophilic and less membrane permeable, which can lead to less bystander effects and less non- specific toxicities compared to conjugates with a cleavable linker. Conjugates with non- cleavable linkers can have greater stability in circulation than conjugates with cleavable linkers. Non-cleavable linkers can include alkylene chains, or can be polymeric, such as, for example, 15 based upon polyalkylene glycol polymers, amide polymers, or can include segments of alkylene chains, polyalkylene glycols and/or amide polymers. The linker can contain a polyethylene glycol segment having from 1 to 6 ethylene glycol units. In some embodiments, -L¹-R¹ or L²-R² comprises a linker that is non-cleavable in vivo. In some embodiments, a trigger element and an immolative unit (IM) together comprise 20 one of the following structures: , 112 15077.006WO1 113 15077.006WO1 TPI-VHL LINKER COMPOUNDS 5 The degrader antibody conjugate (DAC) compositions of the invention are prepared by conjugation of an antibody (Ab) with a target protein binder and VHL ligand linker (TVL) compound. A TVL-VHL linker compound is selected from Formula III: (TPI−Sp−VHL)−L³−Z III 10 or a pharmaceutically acceptable salt, stereoisomer, mixture of stereoisomers, tautomer, or isotopic analog thereof, wherein: TPI is a target protein binder that binds to RIPK2, KRAS, Bcl-XL, or WEE1; VHL ligand has Formula IIIa: 114 15077.006WO1 or a pharmaceutically acceptable salt, stereoisomer, mixture of stereoisomers, tautomer, or isotopic analog thereof, wherein: A is a cyclic structure selected from C₃−C₂₀ carbocyclyl, C₆−C₂₀ aryl, C₁−C₂₀ heteroaryl, 5 and C2-C20 heterocyclyl, each of which are substituted with one or more groups independently selected from H, F, Cl, Br, I, −CN, −NO₂, C₁−C₁₂ alkyl, C₂−C₁₂ alkenyl, C₂−C₁₂ alkynyl, C₁−C₁₂ heteroalkyl, −(C₁−C₁₂ heteroalkyldiyl)−(C₆−C₂₀ aryl), −(C₁−C₁₂ heteroalkyldiyl)−(C₆−C₂₀ aryldiyl)−(C₁−C₁₂ heteroalkyl), −(C₁−C₁₂ heteroalkyldiyl)−(C₆−C₂₀ aryldiyl)−(C₂-C₂₀ heterocyclyl), (C₁−C₆ alkyldiyl)−(C₆−C₂₀ aryl), −(C₁−C₆ alkyldiyl)−NR^aR^b, −(C₁−C₆ 10 alkyldiyl)−OR^a, (C₁−C₆ alkyldiyl)−(C₃−C₂₀ carbocyclyl), (C₁-C₆ alkyldiyl)−(C₂-C₂₀ heterocyclyl), (C1−C6 alkyldiyl)−(C1−C20 heteroaryl), C6−C20 aryl, C3−C20 carbocyclyl, C2−C20 heterocyclyl, C1−C20 heteroaryl, −C(=NH)NH(OH), −C(=NH)NH2, −C(=O)NR^aR^b, −C(=O)NR^a−NR^aR^b, −C(=O)NH(C1-C6 alkyldiyl)−NR^aR^b, −C(=O)OR^a, −NR^aR^b, −NR^aS(O)2R^a, −NO2, −OR^a, −OC(=O)R^a, −SR^a, −S(O)R^a, −S(O)2R^a, −S(O)2NR^a, and −S(O)3H; 15 R^a is independently selected from H, C1−C6 alkyl, phenyl, and benzyl, wherein phenyl and benzyl are optionally substituted with one or more groups independently selected from the group consisting of F, Cl, −CN, C₁−C₁₂ alkyl, C₂−C₁₂ alkenyl, and C₂−C₁₂ alkynyl; or where two R^a groups form a five-, six-, or seven-membered heteroaryl or heterocyclyl ring; 20 R^b is independently selected from H, OH, C1−C6 alkyl, phenyl, and benzyl, wherein phenyl and benzyl are optionally substituted with one or more groups independently selected from the group consisting of F, Cl, −CN, C1−C12 alkyl, C2−C12 alkenyl, and C2−C12 alkynyl; R^c is selected from OH and −SO2F; R^d is selected from H and F; 115 15077.006WO1 Cyc is a ring structure selected from the group consisting of C3−C20 carbocyclyl, C6−C20 aryl, C2-C20 heterocyclyl, C1−C20 heteroaryl, and −(C1−C20 heteroaryldiyl)−(C1−C12 heteroalkyl); n is 0 or 1; 5 R¹ is selected from the group consisting of H, C₁−C₁₂ alkyl, and C₁−C₁₂ heteroalkyl; R² is selected from the group consisting of H, C₁−C₁₂ alkyl, and C₁−C₁₂ heteroalkyl; or where R² forms a five- to ten-membered aryl, carbocyclyl, heteroaryl or heterocyclyl ring with A; X¹ is selected from the group consisting of H, −NHC(=O)−, (C3−C20 carbocyclyl)− 10 C(=O)NH−, C1−C12 heteroalkyl, and C1−C20 heteroaryl; or where X¹ forms a five-, six-, or seven-membered heteroaryl or heterocyclyl ring with R¹; Sp is selected from the group consisting of a bond, O, NH, C1−C12 alkyldiyl, C1−C60 heteroalkyldiyl, C3−C20 carbocyclyldiyl, C2-C20 heterocyclyldiyl, C6-C20 aryldiyl, C1−C40 heteroaryldiyl, −(C3−C20 carbocyclyldiyl)−(C1−C60 heteroalkyldiyl)−, −(C3−C20 15 carbocyclyldiyl)−(C1−C12 alkyldiyl)−, −(C2-C20 heterocyclyldiyl)−(C1−C60 heteroalkyldiyl)−, −(C2-C20 heterocyclyldiyl)−(C1−C12 alkyldiyl)−, −(C2-C20 heterocyclyldiyl)−(C2-C20 heterocyclyldiyl)−, −(C1−C20 heteroaryldiyl)−(C3−C20 carbocyclyldiyl)−(C1−C60 heteroalkyldiyl)−, a solubilizing unit, and combinations thereof, where the solubilizing unit is selected from polyglycine, polysarcosine, polyethyleneoxy (PEG), a glycoside, C1−C60 20 heteroalkyldiyl, and combinations thereof; TPI−Sp−VHL moiety is covalently attached to an antibody linker L³; Z is: where the wavy line is the attachment to L³; 25 one of A, R^a, R^b, Cyc, R¹, R², and X¹ is attached to Sp; one of A, R^a, R^b, Cyc, R¹, R², X¹, and Sp is attached to L³; and each alkyl, alkyldiyl, alkenyl, alkenyldiyl, alkynyl, alkynyldiyl, heteroalkyl, heteroalkyldiyl, aryl, aryldiyl, carbocyclyl, carbocyclyldiyl, heterocyclyl, heterocyclyldiyl, heteroaryl, and heteroaryldiyl is independently and optionally substituted with one or more30 groups selected from F, Cl, Br, I, −CN, −CH₃, −CH₂CH₃, −CH=CH₂, −C^CH, −C^CCH₃, − C^CCH₂NHCH₃, −CH₂CH₂CH₃, −CH(CH₃)₂, −CH₂CH(CH₃)₂, −CH₂OH, −CH₂OCH₃, − 116 15077.006WO1 CH2CH2OH, −C(CH3)2OH, −CH(OH)CH(CH3)2, −C(CH3)2CH2OH, −CH2CH2SO2CH3, − CH2OP(O)(OH)2, −CH2F, −CHF2, −CF3, −CH2CF3, −CH2CHF2, −CH(CH3)CN, −C(CH3)2CN, − CH2CN, −CH2NH2, −CH2NHSO2CH3, −CH2NHCH3, −CH2N(CH3)2, −CO2H, −COCH3, − CO2CH3, −CO2C(CH3)3, −COCH(OH)CH3, −CONH2, −CONHCH3, −CON(CH3)2, − 5 C(CH3)2CONH2, −NH2, −NHCH3, −N(CH3)2, −NHCOCH3, −N(CH3)COCH3, −NHS(O)2CH3, − N(CH3)C(CH3)2CONH2, −N(CH3)CH2CH2S(O)2CH3, −NHC(=NH)H, −NHC(=NH)CH3, − NHC(=NH)NH₂, −NHC(=O)NH₂, −NO₂, =O, −OH, −OCH₃, −OCH₂CH₃, −OCH₂CH₂OCH₃, − OCH₂CH₂OH, −OCH₂CH₂N(CH₃)₂, −OCH₂F, −OCHF₂, −OCF₃, −OP(O)(OH)₂, −S(O)₂N(CH₃)₂, −SCH₃, −S(O)₂CH₃, and −S(O)₃H. 10 An exemplary embodiment of the Formula III compound includes wherein the VHL ligand has Formula IIIb: . An exemplary embodiment of the Formula III compound includes wherein the VHL ligand has Formula IIIc: 15 wherein: R³, R⁴, R⁵, and R⁶ are independently selected from the group consisting of H, F, Cl, Br, I, −CN, C1−C12 alkyl, C2−C12 alkenyl, C2−C12 alkynyl, C1−C12 heteroalkyl, −(C1−C12 heteroalkyldiyl)−(C6−C20 aryl), −(C1−C12 heteroalkyldiyl)−(C6−C20 aryldiyl)−(C1−C12 20 heteroalkyl), −(C₁−C₁₂ heteroalkyldiyl)−(C₆−C₂₀ aryldiyl)−(C₂-C₂₀ heterocyclyl), (C₁−C₆ 117 15077.006WO1 alkyldiyl)−(C6−C20 aryl), −(C1−C6 alkyldiyl)−NR^aR^b, −(C1−C6 alkyldiyl)−OR^a, (C1−C6 alkyldiyl)−(C3−C20 carbocyclyl), (C1-C6 alkyldiyl)−(C2-C20 heterocyclyl), (C1−C6 alkyldiyl)−(C1−C20 heteroaryl), C6−C20 aryl, C3−C20 carbocyclyl, C2−C20 heterocyclyl, C1−C20 heteroaryl, −C(=NH)NH(OH), −C(=NH)NH2, −C(=O)NR^aR^b, −C(=O)NR^a−NR^aR^b, 5 −C(=O)NH(C1-C6 alkyldiyl)−NR^aR^b, −C(=O)OR^a, −NR^aR^b, −NO2, −OR^a, −OC(=O)R^a, −SR^a, −S(O)R^a, −S(O)2R^a, −S(O)2NR^a, and −S(O)3H; R^a is independently selected from H, C₁−C₆ alkyl, phenyl, and benzyl, wherein phenyl and benzyl are optionally substituted with one or more groups independently selected from the group consisting of F, Cl, −CN, C1−C12 alkyl, C2−C12 alkenyl, and C2−C12 alkynyl; 10 or where two R^a groups form a five-, six-, or seven-membered heteroaryl or heterocyclyl group; R^b is independently selected from H, OH, C₁−C₆ alkyl, phenyl, and benzyl, wherein phenyl and benzyl are optionally substituted with one or more groups independently selected from the group consisting of F, Cl, −CN, C₁−C₁₂ alkyl, C₂−C₁₂ alkenyl, and C₂−C₁₂ alkynyl; 15 or where R³ and R⁴ together form a five-membered or six-membered heteroaryl or heterocyclyl group comprising one or more heteroatoms independently selected from N, O, P and S; one of X¹, R¹, R², R³, R⁴, R⁵, R⁶, R^a, R^b, and Cyc is attached to Sp; one of X¹, R¹, R², R³, R⁴, R⁵, R⁶, R^a, R^b, and Cyc is attached to L³; and 20 each alkyl, alkyldiyl, alkenyl, alkenyldiyl, alkynyl, alkynyldiyl, heteroalkyl, heteroalkyldiyl, aryl, aryldiyl, carbocyclyl, carbocyclyldiyl, heterocyclyl, heterocyclyldiyl, heteroaryl, and heteroaryldiyl is independently and optionally substituted with one or more groups selected from F, Cl, Br, I, −CN, −CH3, −CH2CH3, −CH=CH2, −C^CH, −C^CCH3, − CH2CH2CH3, −CH(CH3)2, −CH2CH(CH3)2, −CH2OH, −CH2OCH3, −CH2CH2OH, −C(CH3)2OH, 25 −CH(OH)CH(CH3)2, −C(CH3)2CH2OH, −CH2CH2SO2CH3, −CH2OP(O)(OH)2, −CH2F, −CHF2, −CF3, −CH2CF3, −CH2CHF2, −CH(CH3)CN, −C(CH3)2CN, −CH2CN, −CH2NH2, − CH2NHSO2CH3, −CH2NHCH3, −CH2N(CH3)2, −CO2H, −COCH3, −CO2CH3, −CO2C(CH3)3, − COCH(OH)CH3, −CONH2, −CONHCH3, −CON(CH3)2, −CONHS(O)2N(CH3)2, − C(CH3)2CONH2, −NH2, −NHCH3, −N(CH3)2, −NHCOCH3, −N(CH3)COCH3, −NHS(O)2CH3, −30 N(CH₃)C(CH₃)₂CONH₂, −N(CH₃)CH₂CH₂S(O)₂CH₃, −NHC(=NH)H, −NHC(=NH)CH₃, − NHC(=NH)NH₂, −NHC(=O)NH₂, −NO₂, =O, −OH, −OCH₃, −OCH₂CH₃, −OCH₂CH₂OCH₃, − OCH₂CH₂OH, −OCH₂CH₂N(CH₃)₂, −OCH₂F, −OCHF₂, −OCF₃, −OP(O)(OH)₂, −S(O)₂N(CH₃)₂, −SCH₃, −S(O)₂CH₃, and −S(O)₃H. 118 15077.006WO1 An exemplary embodiment of the Formula III compound includes wherein L³ has the formula: −Str¹−(PEP)_y−(IM)_z− wherein: 5 Str¹ is a stretcher unit covalently attached to Z; PEP is a protease-cleavable, peptide unit covalently attached to Str¹ and IM or the TPI- VHL moiety when y is 1; IM is an immolative unit covalently attached the TPI-VHL moiety when z is 1; y is 0 or 1; and z is 0 or 1. 10 Exemplary target protein binder and VHL ligand linker (TVL) compounds of Tables 2a-d were prepared and characterized according to the Examples herein. Each compound in Tables 2a-d were characterized by mass spectrometry and demonstrated to have the correct parent ion and mass. 119 15077.006WO1 Table 2b1 Bcl-xL-VHL linker compounds (BXVL) 120 15077.006WO1 121 15077.006WO1 122 15077.006WO1 123 15077.006WO1 124 15077.006WO1 125 15077.006WO1 126 15077.006WO1 127 15077.006WO1 128 15077.006WO1 129 15077.006WO1 Table 2b2 Bcl-xL-VHL linker compounds (BXVL) 130 15077.006WO1 131 15077.006WO1 132 15077.006WO1 133 15077.006WO1 134 15077.006WO1 135 15077.006WO1 136 15077.006WO1 137 15077.006WO1 138 15077.006WO1 139 15077.006WO1 140 15077.006WO1 Table 2c KRAS-VHL linker compounds (KVL) 141 15077.006WO1 142 15077.006WO1 143 15077.006WO1 144 15077.006WO1 145 15077.006WO1 146 15077.006WO1 147 15077.006WO1 148 15077.006WO1 149 15077.006WO1 150 15077.006WO1 Table 2d WEE1-VHL linker compounds (WVL) 151 15077.006WO1 152 15077.006WO1 153 15077.006WO1 154 15077.006WO1 155 15077.006WO1 DEGRADER ANTIBODY CONJUGATES The degrader antibody conjugates (DAC) of the invention induce target-specific protein degradation. Tumor targeting brings specificity to minimize off-target effects. 5 The degrader antibody conjugates (DAC) of the invention comprise a target protein binder and VHL ligand moiety (TPI-VHL) covalently attached to an antibody by an antibody linker, wherein the antibody binds to a tumor-associated antigen or cell-surface receptor. An exemplary embodiment of the DAC of Formula I includes wherein the antibody has a modified Fc region. 10 An exemplary embodiment of the DAC of Formula I includes wherein the antibody has Fc mutations selected from: (i) LALAPA (L234A/L235A/P329A); (ii) LALAPG (L234A/L235A/P329G); (iii) LALASKPA (L234A, L235A, S267K, P329A); 15 (iv) LALAPA-YTE (L234A/L235A/P329A-M252Y/S254T/T256E); (v) LALAPG-YTE (L234A/L235A/P329G-M252Y/S254T/T256E); and (vi) LALASKPA-YTE (L234A, L235A, S267K, P329A- M252Y/S254T/T256E), according to EU numbering. An exemplary embodiment of the DAC of Formula I includes wherein the antibody 20 linker is covalently attached to a cysteine amino acid of the antibody. An exemplary embodiment of the DAC of Formula I includes wherein the antibody is a cysteine-engineered antibody. 156 15077.006WO1 An exemplary embodiment of the DAC of Formula I includes wherein the antibody has one engineered cysteine mutation site selected from heavy-chain E152C, S239C, K246C and S375C, numbered according to the EU system. An exemplary embodiment of the DAC of Formula I includes wherein the antibody has 5 two or three engineered cysteine mutation sites selected from heavy-chain E152C, S239C, K246C and S375C, numbered according to the EU system. An exemplary embodiment of the DAC of Formula I includes wherein the cysteine- mutant antibody comprises a heavy chain cysteine mutation in a sequence selected from the group consisting of: 10 according to the EU system. An exemplary embodiment of the DAC of Formula I includes wherein the cysteine- mutant antibody comprising one, two, or three engineered cysteine mutation sites is selected from (i) to (xvi): (i) HC S239C; 15 (ii) HC K246C; (iii) HC S375C; (iv) HC E152C; (v) HC S239C and K246C; (vi) HC S239C and S375C; 20 (vii) HC S239C and E152C; (viii) HC K246C and S375C; (ix) HC K246C and E152C; (x) HC S375C and E152C; (xi) HC S239C, K246C, and S375C; 25 (xii) HC S239C, K246C, and E152C; (xiii) HC S239C, S375C, and E152C; (xiv) HC K246C, S375C, and E152C; (xv) HC S239C, S375C, and E152C; 157 15077.006WO1 (xvi) HC K246C, S375C, and E152C. An exemplary embodiment of the DAC of Formula I includes wherein the target protein binder of the TPI-VHL binds to RIPK2, KRAS, Bcl-XL, or WEE1. Exemplary embodiments of DAC include Formula I: 5 Ab−[L−(TPI−Sp−VHL)]_p I or a pharmaceutically acceptable salt thereof, wherein: Ab is the antibody; L is the antibody linker; 10 TPI−Sp−VHL is a moiety comprising a target protein binder TPI and a VHL ligand wherein the TPI is covalently attached to the VHL ligand by a spacer unit Sp; and p is an integer from 1 to 12. An exemplary embodiment of the DAC of Formula I includes wherein the TPI, the VHL ligand, or the spacer unit Sp is attached to the antibody linker L. 15 An exemplary embodiment of the DAC of Formula I includes wherein the VHL ligand has Formula Ia: wherein A is a cyclic structure selected from C₃−C₂₀ carbocyclyl, C₆−C₂₀ aryl, C₁−C₂₀ heteroaryl, and C₂-C₂₀ heterocyclyl, each of which are substituted with one or more groups 20 independently selected from H, F, Cl, Br, I, −CN, −NO2, C1−C12 alkyl, C2−C12 alkenyl, C2−C12 alkynyl, C₁−C₁₂ heteroalkyl, −(C₁−C₁₂ heteroalkyldiyl)−(C₆−C₂₀ aryl), −(C₁−C₁₂ heteroalkyldiyl)−(C₆−C₂₀ aryldiyl)−(C₁−C₁₂ heteroalkyl), −(C₁−C₁₂ heteroalkyldiyl)−(C₆−C₂₀ aryldiyl)−(C₂-C₂₀ heterocyclyl), (C₁−C₆ alkyldiyl)−(C₆−C₂₀ aryl), −(C₁−C₆ alkyldiyl)−NR^aR^b, −(C₁−C₆ alkyldiyl)−OR^a, (C₁−C₆ alkyldiyl)−(C₃−C₂₀ carbocyclyl), (C₁-C₆ alkyldiyl)−(C₂-C₂₀ 25 heterocyclyl), (C₁−C₆ alkyldiyl)−(C₁−C₂₀ heteroaryl), C₆−C₂₀ aryl, C₃−C₂₀ carbocyclyl, C₂−C₂₀ 158 15077.006WO1 heterocyclyl, C1−C20 heteroaryl, −C(=NH)NH(OH), −C(=NH)NH2, −C(=O)NR^aR^b, −C(=O)NR^a−NR^aR^b, −C(=O)NH(C1-C6 alkyldiyl)−NR^aR^b, −C(=O)OR^a, −NR^aR^b, −NR^aS(O)2R^a, −NO2, −OR^a, −OC(=O)R^a, −SR^a, −S(O)R^a, −S(O)2R^a, −S(O)2NR^a, and −S(O)3H; R^a is independently selected from H, C1−C6 alkyl, phenyl, and benzyl, wherein phenyl 5 and benzyl are optionally substituted with one or more groups independently selected from the group consisting of F, Cl, −CN, C1−C12 alkyl, C2−C12 alkenyl, and C2−C12 alkynyl; or where two R^a groups form a five-, six-, or seven-membered heteroaryl or heterocyclyl ring; R^b is independently selected from H, OH, C₁−C₆ alkyl, phenyl, and benzyl, wherein 10 phenyl and benzyl are optionally substituted with one or more groups independently selected from the group consisting of F, Cl, −CN, C1−C12 alkyl, C2−C12 alkenyl, and C2−C12 alkynyl; R^c is selected from OH and −SO2F; R^d is selected from H and F; Cyc is a ring structure selected from the group consisting of C3−C20 carbocyclyl, C6−C20 15 aryl, C2-C20 heterocyclyl, C1−C20 heteroaryl, and −(C1−C20 heteroaryldiyl)−(C1−C12 heteroalkyl); n is 0 or 1; R¹ is selected from the group consisting of H, C₁−C₁₂ alkyl, and C₁−C₁₂ heteroalkyl; R² is selected from the group consisting of H, C₁−C₁₂ alkyl, and C₁−C₁₂ heteroalkyl, or 20 where R² forms a five- to ten-membered aryl, carbocyclyl, heteroaryl or heterocyclyl ring with A; X¹ is selected from the group consisting of H, −NHC(=O)−, (C3−C20 carbocyclyl)− C(=O)NH−, C1−C12 heteroalkyl, and C1−C20 heteroaryl; or where X¹ forms a five-, six-, or seven-membered heteroaryl or heterocyclyl ring with R¹; 25 Sp is selected from the group consisting of a bond, O, NH, C1−C12 alkyldiyl, C1−C60 heteroalkyldiyl, C3−C20 carbocyclyldiyl, C2-C20 heterocyclyldiyl, C6-C20 aryldiyl, C1−C40 heteroaryldiyl, −(C3−C20 carbocyclyldiyl)−(C1−C60 heteroalkyldiyl)−, −(C3−C20 carbocyclyldiyl)−(C1−C12 alkyldiyl)−, −(C2-C20 heterocyclyldiyl)−(C1−C60 heteroalkyldiyl)−, −(C2-C20 heterocyclyldiyl)−(C1−C12 alkyldiyl)−, −(C2-C20 heterocyclyldiyl)−(C2-C20 30 heterocyclyldiyl)−, −(C1−C20 heteroaryldiyl)−(C3−C20 carbocyclyldiyl)−(C1−C60 heteroalkyldiyl)−, a solubilizing unit, and combinations thereof, where the solubilizing unit is selected from polyglycine, polysarcosine, polyethyleneoxy (PEG), a glycoside, C1−C60 heteroalkyldiyl, and combinations thereof; L is the antibody linker; 159 15077.006WO1 one of A, R^a, R^b, Cyc, R¹, R², and X¹ is attached to Sp; and one of A, R^a, R^b, Cyc, R¹, R², X¹ and Sp is attached to L; each alkyl, alkyldiyl, alkenyl, alkenyldiyl, alkynyl, alkynyldiyl, heteroalkyl, heteroalkyldiyl, aryl, aryldiyl, carbocyclyl, carbocyclyldiyl, heterocyclyl, heterocyclyldiyl, 5 heteroaryl, and heteroaryldiyl is independently and optionally substituted with one or more groups selected from F, Cl, Br, I, −CN, −CH₃, −CH₂CH₃, −CH=CH₂, −C^CH, −C^CCH₃, − C^CCH₂NHCH₃, −CH₂CH₂CH₃, −CH(CH₃)₂, −CH₂CH(CH₃)₂, −CH₂OH, −CH₂OCH₃, − CH₂CH₂OH, −C(CH₃)₂OH, −CH(OH)CH(CH₃)₂, −C(CH₃)₂CH₂OH, −CH₂CH₂SO₂CH₃, − CH₂OP(O)(OH)₂, −CH₂F, −CHF₂, −CF₃, −CH₂CF₃, −CH₂CHF₂, −CH(CH₃)CN, −C(CH₃)₂CN, −10 CH₂CN, −CH₂NH₂, −CH₂NHSO₂CH₃, −CH₂NHCH₃, −CH₂N(CH₃)₂, −CO₂H, −COCH₃, − CO₂CH₃, −CO₂C(CH₃)₃, −COCH(OH)CH₃, −CONH₂, −CONHCH₃, −CON(CH₃)₂, − C(CH3)2CONH2, −NH2, −NHCH3, −N(CH3)2, −NHCOCH3, −N(CH3)COCH3, −NHS(O)2CH3, − N(CH3)C(CH3)2CONH2, −N(CH3)CH2CH2S(O)2CH3, −NHC(=NH)H, −NHC(=NH)CH3, − NHC(=NH)NH2, −NHC(=O)NH2, −NO2, =O, −OH, −OCH3, −OCH2CH3, −OCH2CH2OCH3, − 15 OCH2CH2OH, −OCH2CH2N(CH3)2, −OCH2F, −OCHF2, −OCF3, −OP(O)(OH)2, −S(O)2N(CH3)2, −SCH3, −S(O)2CH3, and −S(O)3H. An exemplary embodiment of the DAC of Formula I includes wherein a solubilizing unit selected from polyglycine, polysarcosine, polyethyleneoxy (PEG), and a glycoside, or combinations thereof is attached to: (1) a phenolic oxygen, or (2) a nitrogen of the VHL ligand 20 moiety. An exemplary embodiment of the DAC of Formula I includes wherein Cyc is a five-, six-, or seven-membered ring structure selected from the group consisting of thiazole, triazole, phenyl, pyridine, pyrazine, pyridazine, and pyrimidine, substituted with one or more groups independently selected from H, F, Cl, Br, I, −CN, −CH3, −CH2CH3, −CH=CH2, −C^CH, −25 C^CCH3, −C^CCH2NHCH3, −CH2CH2CH3, −CH(CH3)2, −CH2CH(CH3)2, −CH2OH, − CH2OCH3, −CH2CH2OH, −C(CH3)2OH, −CH(OH)CH(CH3)2, −C(CH3)2CH2OH, − CH2CH2SO2CH3, −CH2OP(O)(OH)2, −CH2F, −CHF2, −CF3, −CH2CF3, −CH2CHF2, − CH(CH3)CN, −C(CH3)2CN, −CH2CN, −CH2NH2, −CH2NHSO2CH3, −CH2NHCH3, − CH2N(CH3)2, −CO2H, −COCH3, −CO2CH3, −CO2C(CH3)3, −COCH(OH)CH3, −CONH2, −30 CONHCH₃, −CON(CH₃)₂, −C(CH₃)₂CONH₂, −NH₂, −NHCH₃, −N(CH₃)₂, −NHCOCH₃, − N(CH₃)COCH₃, −NHS(O)₂CH₃, −N(CH₃)C(CH₃)₂CONH₂, −N(CH₃)CH₂CH₂S(O)₂CH₃, − NHC(=NH)H, −NHC(=NH)CH₃, −NHC(=NH)NH₂, −NHC(=O)NH₂, −NO₂, =O, −OH, −OCH₃, −OCH₂CH₃, −OCH₂CH₂OCH₃, −OCH₂CH₂OH, −OCH₂CH₂N(CH₃)₂, −OCH₂F, −OCHF₂, − OCF₃, −OP(O)(OH)₂, −S(O)₂N(CH₃)₂, −SCH₃, −S(O)₂CH₃, and −S(O)₃H. 160 15077.006WO1 An exemplary embodiment of the DAC of Formula I includes wherein R¹ is selected from the group consisting of H, C₁−C₁₂ alkyl, and C₁−C₁₂ heteroalkyl. An exemplary embodiment of the DAC of Formula I includes wherein R² is selected from the group consisting of H, C₁−C₁₂ alkyl, and C₁−C₁₂ heteroalkyl. 5 An exemplary embodiment of the DAC of Formula I includes wherein X¹ is C₁−C₂₀ heteroaryldiyl selected from the group consisting of triazole, isoxazole, and oxadiazole. An exemplary embodiment of the DAC of Formula I includes wherein X¹ is − C(=O)NH−. An exemplary embodiment of the DAC of Formula I includes wherein R¹ is selected 10 from H, CH3. An exemplary embodiment of the DAC of Formula I includes wherein R² is selected from H, CH₃, CH₂OH, CH₂OPO₂OH, CH₂COOH, CH₂CH₂COOH, CH₂CH₂CONHS(O)₂CH₃, CH2CH2CONHS(O)2CH2CH2N(CH3)2, and CH2CH2CO(N-methylpiperidyl). An exemplary embodiment of the DAC of Formula I includes wherein Sp is C1−C60 15 heteroalkyldiyl having the formula: −(CH2CH2X²)n−(CH2)m− where X² is independently selected from NH and O, m is an integer from 1 to 5, and n is an integer from 1 to 50. An exemplary embodiment of the DAC of Formula I includes wherein Sp is C₁−C₆₀ heteroalkyldiyl having the formula: −(CH₂CH₂O)_n−(CH₂)_m− where m is an integer from 1 to 5, and n is an integer from 1 to 50. 20 An exemplary embodiment of the DAC of Formula I includes wherein Sp is −(C₃−C₂₀ carbocyclyldiyl)−(C₁−C₆₀ heteroalkyldiyl)− . An exemplary embodiment of the DAC of Formula I includes wherein C₃−C₂₀ carbocyclyldiyl is an optionally substituted adamantyl group. An exemplary embodiment of the DAC of Formula I includes wherein Sp has a formula 25 selected from the group consisting of: 161 15077.006WO1 162 15077.006WO1 where the wavy lines indicate the points of attachment to the target protein binder TPI and to the VHL ligand. An exemplary embodiment of the DAC of Formula I includes wherein the VHL ligand has Formula Ib: 5 . An exemplary embodiment of the DAC of Formula I includes wherein the VHL ligand has Formula Ic: wherein: 10 R³, R⁴, R⁵, and R⁶ are independently selected from the group consisting of H, F, Cl, Br, I, −CN, C1−C12 alkyl, C2−C12 alkenyl, C2−C12 alkynyl, C1−C12 heteroalkyl, −(C1−C12 heteroalkyldiyl)−(C6−C20 aryl), −(C1−C12 heteroalkyldiyl)−(C6−C20 aryldiyl)−(C1−C12 heteroalkyl), −(C1−C12 heteroalkyldiyl)−(C6−C20 aryldiyl)−(C2-C20 heterocyclyl), (C1−C6 alkyldiyl)−(C₆−C₂₀ aryl), −(C₁−C₆ alkyldiyl)−NR^aR^b, −(C₁−C₆ alkyldiyl)−OR^a, (C₁−C₆ 15 alkyldiyl)−(C₃−C₂₀ carbocyclyl), (C₁-C₆ alkyldiyl)−(C₂-C₂₀ heterocyclyl), (C₁−C₆ alkyldiyl)−(C₁−C₂₀ heteroaryl), C₆−C₂₀ aryl, C₃−C₂₀ carbocyclyl, C₂−C₂₀ heterocyclyl, C₁−C₂₀ heteroaryl, −C(=NH)NH(OH), −C(=NH)NH₂, −C(=O)NR^aR^b, −C(=O)NR^a−NR^aR^b, −C(=O)NH(C₁-C₆ alkyldiyl)−NR^aR^b, −C(=O)OR^a, −NR^aR^b, −NO₂, −OR^a, −OC(=O)R^a, −SR^a, −S(O)R^a, −S(O)₂R^a, −S(O)₂NR^a, and −S(O)₃H; 163 15077.006WO1 R^a is independently selected from H, C1−C6 alkyl, phenyl, and benzyl, wherein phenyl and benzyl are optionally substituted with one or more groups independently selected from the group consisting of F, Cl, −CN, C₁−C₁₂ alkyl, C₂−C₁₂ alkenyl, and C₂−C₁₂ alkynyl; or where two R^a groups form a five-, six-, or seven-membered heteroaryl or heterocyclyl 5 group; R^b is independently selected from H, OH, C1−C6 alkyl, phenyl, and benzyl, wherein phenyl and benzyl are optionally substituted with one or more groups independently selected from the group consisting of F, Cl, −CN, C1−C12 alkyl, C2−C12 alkenyl, and C2−C12 alkynyl; or where R³ and R⁴ together form a five-membered or six-membered heteroaryl or 10 heterocyclyl group comprising one or more heteroatoms independently selected from N, O, P and S; one of X¹, R¹, R², R³, R⁴, R⁵, R⁶, R^a, R^b, and Cyc is attached to the spacer unit; one of X¹, R¹, R², R³, R⁴, R⁵, R⁶, R^a, R^b, Cyc, and Sp is attached to L; and each alkyl, alkyldiyl, alkenyl, alkenyldiyl, alkynyl, alkynyldiyl, heteroalkyl, 15 heteroalkyldiyl, aryl, aryldiyl, carbocyclyl, carbocyclyldiyl, heterocyclyl, heterocyclyldiyl, heteroaryl, and heteroaryldiyl is independently and optionally substituted with one or more groups selected from F, Cl, Br, I, −CN, −CH₃, −CH₂CH₃, −CH=CH₂, −C^CH, −C^CCH₃, − CH₂CH₂CH₃, −CH(CH₃)₂, −CH₂CH(CH₃)₂, −CH₂OH, −CH₂OCH₃, −CH₂CH₂OH, −C(CH₃)₂OH, −CH(OH)CH(CH₃)₂, −C(CH₃)₂CH₂OH, −CH₂CH₂SO₂CH₃, −CH₂OP(O)(OH)₂, −CH₂F, −CHF₂,20 −CF3, −CH2CF3, −CH2CHF2, −CH(CH3)CN, −C(CH3)2CN, −CH2CN, −CH2NH2, − CH2NHSO2CH3, −CH2NHCH3, −CH2N(CH3)2, −CO2H, −COCH3, −CO2CH3, −CO2C(CH3)3, − COCH(OH)CH3, −CONH2, −CONHCH3, −CON(CH3)2, −CONHS(O)2N(CH3)2, − C(CH3)2CONH2, −NH2, −NHCH3, −N(CH3)2, −NHCOCH3, −N(CH3)COCH3, −NHS(O)2CH3, − N(CH3)C(CH3)2CONH2, −N(CH3)CH2CH2S(O)2CH3, −NHC(=NH)H, −NHC(=NH)CH3, −25 NHC(=NH)NH2, −NHC(=O)NH2, −NO2, =O, −OH, −OCH3, −OCH2CH3, −OCH2CH2OCH3, − OCH2CH2OH, −OCH2CH2N(CH3)2, −OCH2F, −OCHF2, −OCF3, −OP(O)(OH)2, −S(O)2N(CH3)2, −SCH3, −S(O)2CH3, and −S(O)3H. An exemplary embodiment of the DAC of Formula I includes wherein X¹ forms a five-, six-, or seven-membered heteroaryl or heterocyclyl ring with R¹. 30 An exemplary embodiment of the DAC of Formula I includes wherein the target protein binder TPI binds to Bcl-xL and is selected from the structures: 164 15077.006WO1 wherein: R¹⁰ is a bicyclic C1−C20 heteroaryl substituted with one or more groups independently selected from H, F, Cl, −CH3, −CN, −NO2, −OH, and −OCH3; 5 R¹¹ is selected from H, −(C6−C20 aryl), −(C1−C20 heteroaryldiyl)−(C1-C12 alkyl), −(C₁−C₂₀ heteroaryldiyl)−(C₁-C₁₂ alkyldiyl)−(C₃−C₂₀ carbocyclyldiyl)−(C₁−C₆₀ heteroalkyl), and −(C₁−C₂₀ heteroaryldiyl)−(C₃−C₂₀ carbocyclyldiyl)−(C₁−C₆₀ heteroalkyl); R¹² is selected from −(C₁−C₂₀ heteroaryldiyl)−(C₁-C₁₂ alkyldiyl)−(C₃−C₂₀ carbocyclyldiyl)−(C₁−C₆₀ heteroalkyldiyl)−*; 10 R¹³ is selected from −(C₁−C₆₀ heteroalkyldiyl)−*; and the asterisk * indicates the attachment to the spacer Sp. An exemplary embodiment of the DAC of Formula I includes wherein R¹⁰ is selected from: 165 15077.006WO1 , substituted with one or more groups independently selected from H, F, Cl, −CH3, −CN, −NO2, −OH, and −OCH3. An exemplary embodiment of the DAC of Formula I includes wherein R¹⁰ is: 5 . An exemplary embodiment of the DAC of Formula I includes wherein the target protein binder TPI binds to KRAS and has Formula Id: wherein: 10 X², X³, and X⁴ are independently selected from the group consisting of C(R^aR^b), O, S, and NR^a, or where: (i) X² and X⁴, (ii) X³ and X⁴, or (iii) X² and X³ can form a 3-6 membered carbocyclic ring; the dotted line indicates an optional double bond; 166 15077.006WO1 Y¹ is selected from the group consisting of CH, N, O, S; Y² is selected from the group consisting of N, O, S; Y³ is selected from the group consisting of N, O, S; R¹⁴ is selected from the group consisting of H, C1−C6 alkyl, C2−C12 alkenyl, and C2−C12 5 alkynyl, substituted with one or more groups independently selected from the group consisting of H, F, Cl, −CN, −NO2, −OH, and −CN; or where R¹⁴ and Y¹ form a six-membered ring, optionally-substituted with one or more groups selected from F, spiro-cyclopropyl, C₁−C₁₂ heteroalkyl, and C₁−C₁₂ alkyl; R¹⁵ is selected from the group consisting of −O−(C₁−C₁₂ heteroalkyldiyl)−(C₂-C₂₀ 10 heterocyclyl), −O−(C₁−C₁₂ alkyldiyl)−(C₂-C₂₀ heterocyclyl), −(C₁−C₁₂ heteroalkyldiyl)−(C₂-C₂₀ heterocyclyl), −(C₁−C₁₂ alkyldiyl)−(C₂-C₂₀ heterocyclyl), −(C₂-C₂₀ heterocyclyl)−(C₂-C₂₀ heterocyclyl), and C2-C20 heterocyclyl, where at least one of the heterocyclyl ring atoms comprise one or more basic nitrogens, and the heterocyclyl group is substituted with one or more groups independently selected from H, F, Cl, =CH2, C1−C6 alkyl, −C(=NH)NH(OH), 15 −C(=NH)NH₂, −C(=O)NR^aR^b, −C(=O)NR^a−NR^aR^b, −C(=O)NH−(C₁-C₆ alkyldiyl)−NR^aR^b, −C(=O)OR^a, −NR^aR^b, −NO₂, −OR^a, −OC(=O)R^a, −SR^a, −S(O)R^a, −S(O)₂R^a, −S(O)₂NR^a, and −S(O)₃H; R^a is independently selected from H, C₁−C₆ alkyl, phenyl, and benzyl, wherein phenyl and benzyl are substituted with one or more groups independently selected from the group 20 consisting of H, F, Cl, −CN, −NO2, −OH, C1−C12 alkyl, C2−C12 alkenyl, and C2−C12 alkynyl; or where two R^a groups form a five-, six-, or seven-membered heteroaryl or heterocyclyl ring; R^b is independently selected from H, OH, C₁−C₆ alkyl, phenyl, and benzyl, wherein phenyl and benzyl are substituted with one or more groups independently selected from the 25 group consisting of H, F, Cl, −CN, C₁−C₁₂ alkyl, C₂−C₁₂ alkenyl, and C₂−C₁₂ alkynyl; R¹⁶ is selected from the group consisting of −(C₁−C₂₀ heteroaryldiyl)−*, −(C₁−C₂₀ heteroaryldiyl)−C(=O)NR^a−*, −(C₁−C₂₀ heteroaryldiyl)−(C₁−C₆ alkyldiyl)−C(=O)NR^a−*, −(C₁−C₂₀ heteroaryldiyl)−(C₁−C₆ alkyldiyl)−*, −(C₆−C₂₀ aryldiyl)−*, −(C₆−C₂₀ aryldiyl)−C(=O)NR^a−*, −(C₆−C₂₀ aryldiyl)−(C₁−C₆ alkyldiyl)−C(=O)NR^a−*, −(C₁−C₂₀ 30 heteroaryldiyl)−(C₁−C₆ alkyldiyl)−(C₁−C₂₀ heteroaryldiyl)−*, −(C₁−C₂₀ heteroaryldiyl)−(C₁−C₆ heteroalkyldiyl)−(C1−C20 heteroaryldiyl)−*, −(C1−C20 heteroaryldiyl)−(C1−C6 alkyldiyl)−(C1−C20 heterocyclyl)−*, −(C1−C20 heteroaryldiyl)−(C1−C6 heteroalkyldiyl)−(C1−C20 167 15077.006WO1 heterocyclyl)−*, −O−(C1−C20 heteroaryldiyl)−*, and −O−(C1−C20 heteroaryldiyl)−C(=O)NR^a−*; where the asterisk is the attachment site to the spacer unit Sp. An exemplary embodiment of the DAC of Formula I includes wherein the target protein 5 binder TPI has a formula selected from the group consisting of: where the wavy line indicates the point of attachment to the spacer unit Sp. An exemplary embodiment of the DAC of Formula I includes wherein the target protein binder TPI binds to WEE1 and has Formula Ie: 10 wherein: R¹⁷ is selected from the group consisting of C1−C6 alkyl, C2−C6 alkenyl, and C2−C6 alkynyl, each of which are substituted with one or more groups independently selected from H, F, Cl, −CH₃, −CN, −NO₂, −OH, and −OCH₃ R¹⁸ is C₁−C₂₀ heteroaryl substituted with one or more groups independently selected 15 from H, F, Cl, −CN, −NO2, −OH, −OCH3, C1−C6 alkyl, and C1−C20 heteroalkyl; R¹⁹ is selected from the group consisting of H and C1−C6 alkyl; and R²⁰ is C1−C6 alkyldiyl attached to the spacer Sp; or where R¹⁹ and R²⁰ form a five-, six-, or seven-membered heterocycle attached to the spacer Sp; 20 where each alkyl, alkyldiyl, alkenyl, alkynyl, alkynyldiyl, heteroalkyl, heterocyclyl, and heteroaryl is substituted with one or more groups selected from H, F, Cl, Br, I, −CN, −CH3, − CH2CH3, −CH=CH2, −C^CH, −C^CCH3, −C^CCH2NHCH3, −CH2CH2CH3, −CH(CH3)2, − 168 15077.006WO1 CH2CH(CH3)2, −CH2OH, −CH2OCH3, −CH2CH2OH, −C(CH3)2OH, −CH(OH)CH(CH3)2, − C(CH3)2CH2OH, −CH2CH2SO2CH3, −CH2OP(O)(OH)2, −CH2F, −CHF2, −CF3, −CH2CF3, − CH2CHF2, −CH(CH3)CN, −C(CH3)2CN, −CH2CN, −CH2NH2, −CH2NHSO2CH3, −CH2NHCH3, −CH2N(CH3)2, −CO2H, −COCH3, −CO2CH3, −CO2C(CH3)3, −COCH(OH)CH3, −CONH2, − 5 CONHCH3, −CON(CH3)2, −C(CH3)2CONH2, −NH2, −NHCH3, −N(CH3)2, −NHCOCH3, − N(CH3)COCH3, −NHS(O)2CH3, −N(CH3)C(CH3)2CONH2, −N(CH3)CH2CH2S(O)2CH3, − NHC(=NH)H, −NHC(=NH)CH₃, −NHC(=NH)NH₂, −NHC(=O)NH₂, −NO₂, =O, −OH, −OCH₃, −OCH₂CH₃, −OCH₂CH₂OCH₃, −OCH₂CH₂OH, −OCH₂CH₂N(CH₃)₂, −OCH₂F, −OCHF₂, − OCF₃, −OP(O)(OH)₂, −S(O)₂N(CH₃)₂, −SCH₃, −S(O)₂CH₃, and −S(O)₃H. 10 An exemplary embodiment of the DAC of Formula I includes wherein R¹⁷ is allyl. An exemplary embodiment of the DAC of Formula I includes wherein R¹⁸ is pyridyl substituted with C1−C20 heteroalkyl. An exemplary embodiment of the DAC of Formula I includes wherein R¹⁹ and R²⁰ form a piperidyl ring attached to the spacer Sp. 15 An exemplary embodiment of the DAC of Formula I includes wherein Formula Ie has the structure: where the asterisk is the attachment site to the spacer unit Sp. An exemplary embodiment of the DAC of Formula I includes wherein the target protein 20 binder TPI has a formula selected from the group consisting of: 169 15077.006WO1 170 15077.006WO1 171 15077.006WO1 where the wavy line indicates the point of attachment to the spacer unit Sp. An exemplary embodiment of the DAC of Formula I includes wherein L comprises a solubilizing unit selected from polyglycine, polysarcosine, polyethyleneoxy (PEG), and a glycoside, or combinations thereof. 5 An exemplary embodiment of the DAC of Formula I includes wherein the solubilizing unit is monovalent and the terminus of the solubilizing unit is a group selected from an amino acid, amino, hydroxyl, hydrogen, carboxylic acid, glycerol, or a sugar such as pentaerythritol, maltitol, sorbitol, xylitol, erythritol, isomalt, or combinations thereof. An exemplary embodiment of the DAC of Formula I includes wherein the solubilizing 10 unit and the terminus of the solubilizing unit covalently attached to Str are selected from the structures: ; 172 15077.006WO1 5 ; 173 15077.006WO1 5 wherein * indicates the point of attachment to Str. An exemplary embodiment of the DAC of Formula I includes wherein L has the structure: wherein: 174 15077.006WO1 L¹ is selected from a bond, C1−C12 alkyldiyl, and C1−C40 heteroalkyldiyl; L^1a is selected from an amino acid, amino, hydroxyl, halide, hydrogen, carboxylic acid, glycerol, or a sugar such as pentaerythritol, maltitol, sorbitol, xylitol, erythritol, isomalt, or combinations thereof; 5 o is 0, 1, or 2; L² is selected from a bond, C1−C12 alkyldiyl, and C1−C40 heteroalkyldiyl; * indicates the point of attachment to a cysteine thiol of Ab; and ** indicates the point of attachment to the TPI-VHL moiety. An exemplary embodiment of the DAC of Formula I includes wherein L¹ is a bond and 10 L^1a is F. An exemplary embodiment of the DAC of Formula I includes wherein L has the formula: −Str−(PEP)_y−(IM)_z− wherein: Str is a stretcher unit covalently attached to the antibody; 15 PEP is a protease-cleavable, peptide unit covalently attached to Str and IM or the TPI- VHL moiety; IM is an immolative unit covalently attached to the TPI-VHL moiety; y is 0 or 1; and z is 0 or 1. An exemplary embodiment of the DAC of Formula I includes wherein L is a branched 20 linker and Str is covalently attached to: (i) the antibody; and (ii) a solubilizing unit comprising a group selected from polyglycine, polysarcosine, polyethyleneoxy (PEG), and a glycoside, or combinations thereof, wherein the terminus of the solubilizing unit is a group selected from an amino acid, amino, hydroxyl, hydrogen, carboxylic acid, glycerol, or a sugar such as pentaerythritol, maltitol, sorbitol, xylitol, erythritol, isomalt, or combinations thereof. 25 An exemplary embodiment of the DAC of Formula I includes wherein L is a branched linker and PEP is covalently attached to: (i) Str and IM or the TPI-VHL moiety; and (ii) a solubilizing unit comprises a group selected from polyglycine, polysarcosine, polyethyleneoxy (PEG), and a glycoside, or combinations thereof, wherein the terminus of the solubilizing unit is a group selected from an amino acid, amino, hydroxyl, hydrogen, carboxylic acid, glycerol, or a 30 sugar such as pentaerythritol, maltitol, sorbitol, xylitol, erythritol, isomalt, or combinations thereof. An exemplary embodiment of the DAC of Formula I includes wherein L is a branched linker and IM is covalently attached to: (i) the TPI-VHL moiety; and (ii) a solubilizing unit comprises a group selected from polyglycine, polysarcosine, polyethyleneoxy (PEG), and a 175 15077.006WO1 glycoside, or combinations thereof, wherein the terminus of the solubilizing unit is a group selected from an amino acid, amino, hydroxyl, hydrogen, carboxylic acid, glycerol, or a sugar such as pentaerythritol, maltitol, sorbitol, xylitol, erythritol, isomalt, or combinations thereof. An exemplary embodiment of the DAC of Formula I includes wherein Str has the 5 structure: wherein: * indicates the point of attachment to a cysteine thiol of Ab; ** indicates the point of attachment to PEP or to the TPI-VHL moiety; 10 R^e is selected from the group consisting of C₁-C₁₂ alkyldiyl, C₁-C₁₂ alkyldiyl-C(=O), C₁- C12 alkyldiyl−NH, (CH2CH2O)r, (CH2CH2O)r−C(=O), (CH2CH2O)r-CH2, C1−C12 heteroalkyldiyl, C6−C20 aryldiyl, (C6−C20 aryldiyl)−(C1−C12 alkyldiyl), and (C6−C20 aryldiyl)− (C1−C12 heteroalkyldiyl); r is an integer ranging from 1 to 10; and 15 alkyldiyl, heteroalkyldiyl, and aryldiyl are independently and optionally substituted with one or more groups selected from F, Cl, −CN, −NH₂, −CH₂NH₂, −OH, −OCH₃, −OCH₂CH₃, − OCH₂CH₂OCH₃, −OCH₂CH₂OH, −OCH₂CH₂N(CH₃)₂, −OCH₂F, −OCHF₂, −OCF₃, − OP(O)(OH)₂, −S(O)₂N(CH₃)₂, −SCH₃, −S(O)₂CH₃, −S(O)₃H, and a solubilizing unit. An exemplary embodiment of the DAC of Formula I includes wherein R^e is selected 20 from −(CH₂)₅−, and −CH₂CH₂−. An exemplary embodiment of the DAC of Formula I includes wherein R^e is selected from C6−C20 aryldiyl, (C6−C20 aryldiyl)−(C1−C12 alkyldiyl), and (C6−C20 aryldiyl)−(C1−C12 heteroalkyldiyl). An exemplary embodiment of the DAC of Formula I includes wherein Str is selected 25 from the structure: 176 15077.006WO1 wherein: L¹ is selected from a bond, C₁−C₁₂ alkyldiyl, and C₁−C₄₀ heteroalkyldiyl; L^1a is selected from an amino acid, amino, hydroxyl, hydrogen, carboxylic acid, glycerol, or a sugar such as pentaerythritol, maltitol, sorbitol, xylitol, erythritol, isomalt, or combinations 5 thereof; o is 0, 1, or 2; L² is selected from a bond, C₁−C₁₂ alkyldiyl, and C₁−C₄₀ heteroalkyldiyl; * indicates the point of attachment to a cysteine thiol of Ab; and ** indicates the point of attachment to PEP or to the TPI-VHL moiety. 10 An exemplary embodiment of the DAC of Formula I includes wherein L¹ or L² comprise a solubilizing unit selected from polyglycine, polysarcosine, polyethyleneoxy (PEG), and a glycoside, or combinations thereof. An exemplary embodiment of the DAC of Formula I includes wherein L¹ or L² is selected from (N(CH3)CH2C(=O))q, (N(CH3)CH2CH2C(=O))q, 15 N(CH₃)CH₂CH₂OCH₂CH₂C(=O))_q, (CH₂CH₂O)_q, (CH₂CH₂O)_q−C(=O), and (CH₂CH₂O)_q-CH₂, where q is an integer from 2 to 20. An exemplary embodiment of the DAC of Formula I is selected from formulas: wherein * indicates the point of attachment to a cysteine thiol of Ab and ** indicates the 20 point of attachment to the TPI-VHL moiety. 177 15077.006WO1 An exemplary embodiment of the DAC of Formula I includes wherein n is 1, m is 1, and PEP-IM has the formula: wherein * indicates the point of attachment to Str and ** indicates the point of 5 attachment to the TPI-VHL moiety; AA is independently selected from a natural or unnatural amino acid side chain, or one or more of AA, and an adjacent nitrogen atom form a 5-membered ring proline amino acid; Cyc¹ is selected from C6-C20 aryldiyl and C1-C20 heteroaryldiyl, optionally substituted with one or more groups selected from F, Cl, NO2, −OH, −OCH3, and glucuronic acid having 10 the structure: R⁷ is selected from the group consisting of −CH(R⁸)O−, −CH₂−, −CH₂N(R⁸)CH(R⁸)−, − CH(R⁸)OC(=O)−, −CH(R⁸)OC(=O)N(R⁸)CH(R⁸)−, −CH(R⁸)OP(=O)2OCH(R⁸)−, and − CH(R⁸)OC(=O)N(R⁸)−(C₁-C₆ alkyldiyl)−N(R⁸)C(=O)OCH(R⁸)−; 15 R⁸ is selected from H, C₁-C₆ alkyl, C(=O)−C₁-C₆ alkyl, and −C(=O)N(R⁹)₂; R⁹ is independently selected from the group consisting of H, C₁-C₁₂ alkyl, and − (CH₂CH₂O)_n−(CH₂)_m−OH, where m is an integer from 1 to 5, and n is an integer from 2 to 50, or two R⁹ groups together form a 5- or 6-membered heterocyclyl ring; y¹ is an integer from 2 to 12; and 20 z¹ is 0 or 1. An exemplary embodiment of the DAC of Formula I includes wherein IM is selected from the formulae: 178 15077.006WO1 * indicates the point of attachment to PEP; and 5 ** indicates the point of attachment to the TPI-VHL moiety. The invention includes all reasonable combinations, and permutations of the features, of the Formula I embodiments. In certain embodiments, the degrader antibody conjugates (DAC) of the invention include those with anti-cancer activity. The DAC selectively deliver an effective dose of a TPI- 10 VHL drug to tumor tissue or hematopoietic cell, whereby greater selectivity (i.e., a lower 179 15077.006WO1 efficacious dose) may be achieved while increasing the therapeutic index (“therapeutic window”) relative to unconjugated TPI-VHL drug. Drug loading in Tables 3a-d is represented as DAR, the number of TPI-VHL moieties per antibody in an DAC of Formula I. Drug TPI-VHLloading may range from 1 to about 8 drug 5 moieties (D) per antibody. DAC of Formula I include mixtures or collections of antibodies conjugated with a range of TPI-VHL drug moieties, from 1 to about 8. In some embodiments, the number of TPI-VHL drug moieties that can be conjugated to an antibody is limited by the number of reactive or available amino acid side chain residues such as lysine and cysteine. In some embodiments, free cysteine residues are introduced into the antibody amino acid sequence 10 by the methods described herein. In such aspects, p may be 1, 2, 3, 4, 5, 6, 7, or 8, and ranges thereof, such as from 1 to 8 or from 2 to 5. Exemplary DAC of Formula I include, but are not limited to, antibodies that have 1, 2, 3, or 4 engineered cysteine amino acids (Lyon, R. et al. (2012) Methods in Enzym.502:123-138). In some embodiments, one or more free cysteine residues are already present in an antibody forming intra-chain and inter-chain disulfide bonds 15 (native disulfide groups), without the use of engineering, in which case the existing free, reduced cysteine residues may be used to conjugate the antibody to a drug. In some embodiments, an antibody is exposed to reducing conditions prior to conjugation of the antibody in order to generate one or more free cysteine residues. For some antibody conjugates, p may be limited by the number of attachment sites on the 20 antibody. For example, where the attachment is a cysteine thiol, as in certain exemplary embodiments described herein, an antibody may have only one or a limited number of cysteine thiol groups, or may have only one or a limited number of sufficiently reactive thiol groups, to which the drug may be attached. In other embodiments, one or more lysine amino groups in the antibody may be available and reactive for conjugation with a TVL compound of Formula III. In 25 certain embodiments, higher drug loading, e.g. p >5, may cause aggregation, insolubility, toxicity, or loss of cellular permeability of certain antibody-drug conjugates. In certain embodiments, the average drug loading for an antibody conjugate ranges from 1 to about 8; from about 2 to about 6; or from about 3 to about 5. In certain embodiments, an antibody is subjected to denaturing conditions to reveal reactive nucleophilic groups such as lysine or 30 cysteine. The loading (drug/antibody ratio) of an antibody conjugate may be controlled in different ways, and for example, by: (i) limiting the molar excess of the TVL intermediate compound relative to antibody, (ii) limiting the conjugation reaction time or temperature, and (iii) partial or limiting reductive denaturing conditions for optimized antibody reactivity. 180 15077.006WO1 It is to be understood that where more than one nucleophilic group of the antibody reacts with a TVL, then the resulting product is a mixture of antibody conjugate compounds with a distribution of one or more TPI-VHL drug moieties attached to an antibody. The average number of drugs per antibody may be calculated from the mixture by a dual ELISA antibody 5 assay, which is specific for antibody and specific for the drug. Individual DAC molecules may be identified in the mixture by mass spectroscopy and separated by HPLC, for example hydrophobic interaction chromatography, HIC (McDonagh et al. (2006) Prot. Engr. Design & Selection 19(7):299-307; Hamblett et al. (2004) Clin. Cancer Res.10:7063-7070; Hamblett, K.J., et al. “Effect of drug loading on the pharmacology, pharmacokinetics, and toxicity of an 10 anti-CD30 antibody-drug conjugate,” Abstract No.624, American Association for Cancer Research, 2004 Annual Meeting, March 27-31, 2004, Proceedings of the AACR, Volume 45, March 2004; Alley, S.C., et al. “Controlling the location of drug attachment in antibody-drug conjugates,” Abstract No.627, American Association for Cancer Research, 2004 Annual Meeting, March 27-31, 2004, Proceedings of the AACR, Volume 45, March 2004). In certain 15 embodiments, a homogeneous antibody conjugate with a single loading value may be isolated from the conjugation mixture by electrophoresis or chromatography. Exemplary embodiments of the degrader antibody conjugates (DAC) of Formula I are compiled in Tables 3a-d. Assessment of DAC biological activity and other properties may be conducted according to the methods of Examples 103-104. In some instances, the Cell 20 proliferation dose response IC50 value is an average of repetitive assays. Certain exemplary DAC were tested for their effects in inhibiting cellular proliferation, including CAL51; WSU- DLCL2, NCI-N87 and SKBR3 cell lines. CAL51 is a human breast adenocarcinoma cell line with triple-negative status for expression of estrogen, progesterone and HER2 receptors. WSU- DLCL2 is a human B-Cell non-Hodgkin lymphoma cell line that expresses high levels of 25 CD22. NCI-N87 is a human epithelial cell line established from a gastric carcinoma; SKBR3 is a human epithelial cell line established from a breast adenocarcinoma; both NCI-N87 and SKBR3 cell lines express high levels of HER2 receptor. 181 15077.006WO1 Table 3a RIPK2-VHL Degrader Antibody Conjugates (DAC-RV) Table 3b1 BCL-xL-VHL Degrader Antibody Conjugates (DAC-BXV) 182 15077.006WO1 Table 3b2 BCL-xL-VHL Degrader Antibody Conjugates (DAC-BXV) 183 15077.006WO1 184 15077.006WO1 185 15077.006WO1 Table 3c KRAS-VHL Degrader Antibody Conjugates (DAC-KV) 186 15077.006WO1 187 15077.006WO1 Table 3d1 WEE1-VHL Degrader Antibody Conjugates (DAC-WV) Table 3d2 WEE1-VHL Degrader Antibody Conjugates (DAC-WV) 188 15077.006WO1 189 15077.006WO1 PHARMACEUTICAL COMPOSITIONS OF DEGRADER ANTIBODY CONJUGATES The invention provides a composition, e.g., a pharmaceutically or pharmacologically acceptable composition or formulation, comprising a TPI-VHL degrader antibody conjugate 5 (DAC) composition of the invention as described herein and a pharmaceutically acceptable diluent, vehicle, carrier or excipient. The degrader antibody conjugate (DAC) composition can be the same or different in the pharmaceutical composition, i.e., the composition can comprise DAC that have the same number of TPI-VHL moieties linked to the same positions on the antibody and/or DAC that have 10 the same number of TPI-VHL moieties linked to different positions on the antibody, that have different numbers of TPI-VHL moieties linked to the same positions on the antibody, or that have different numbers of TPI-VHL moieties linked to different positions on the antibody. In an exemplary embodiment, a pharmaceutical composition comprises a mixture of the DAC, wherein the average drug TPI-VHL loading per antibody in the mixture of antibody conjugate 15 compounds is about 2 to about 8. An DAC of the invention can have an average TPI-VHL to antibody ratio (DAR) of about 0.4 to about 10. A skilled artisan will recognize that the number of IG moieties conjugated to the antibody may vary amongst DAC in a composition comprising multiple DAC of the invention and thus the DAR can be measured as an average which may be referred to as the drug 20 to antibody ratio (DAR) which can be assessed by any suitable means, many of which are known in the art. The average number of TPI-VHL moieties per antibody (DAR) in preparations of DAC from conjugation reactions may be characterized by conventional means such as mass spectrometry, ELISA assay, and HPLC. The quantitative distribution of DAC in a composition in terms of p may also be determined. In certain instances, separation, purification, and 25 characterization of homogeneous DAC where p is a certain value from antibody drug conjugates (ADC) with other drug loadings may be achieved by purification means such as reverse phase HPLC or electrophoresis. A TPI-VHL degrader antibody conjugate (DAC) composition can be formulated for parenteral administration, such as intradermal, subcutaneous, intramuscular (IM), or intravenous 30 (IV) injections, infusion, or administration into a body cavity or lumen of an organ. Alternatively, the DAC as a pharmaceutical composition can be injected into otherwise placed into a specific site of the body, such as a tumor. Compositions for injection will commonly comprise a solution of the DAC dissolved in a pharmaceutically acceptable carrier. Among the acceptable vehicles and solvents that can be employed are water and an isotonic solution of one 190 15077.006WO1 or more salts such as sodium chloride, e.g., Ringer's solution. In addition, sterile fixed oils can conventionally be employed as a solvent or suspending medium. For this purpose, any bland fixed oil can be employed, including synthetic monoglycerides or diglycerides. In addition, fatty acids such as oleic acid can likewise be used in the preparation of injectables. These 5 pharmaceutical compositions desirably are sterile and generally free of undesirable matter. These pharmaceutical compositions can be made sterilized by conventional, well known sterilization techniques. The pharmaceutical compositions can contain pharmaceutically acceptable auxiliary substances as required to approximate physiological conditions such as pH adjusting and buffering agents, toxicity adjusting agents, e.g., sodium acetate, sodium chloride, 10 potassium chloride, calcium chloride, sodium lactate and the like. The pharmaceutical composition may contain any suitable concentration of the DAC . The concentration of the DAC in the pharmaceutical composition can vary widely, and will be selected primarily based on fluid volumes, viscosities, body weight, and the like, in accordance with the particular mode of administration selected and the patient's needs. In certain 15 embodiments, the concentration of DAC in a solution formulation for injection will range from about 0.1% (w/w) to about 10% (w/w). METHODS OF TREATING CANCER WITH DEGRADER ANTIBODY CONJUGATES By inducing target-specific degradation of tumor-associated proteins and conferring specificity to minimize off-target toxicity effects, the TPI-VHL antibody conjugate (DAC) 20 compositions of the invention may be useful in the treatment of diseases and disorders such as cancer. The DAC direct a tumor-associated antigen-binding antibody to a cell that expresses the antigen and deliver a protein-degrading moiety to the target cell. A target protein such as RIPK2, KRAS, Bcl-XL, or WEE1 is ubiquitinated and subsequently degraded. The invention provides a method for treating cancer with a pharmaceutical composition 25 of the DAC. The method includes administering a therapeutically effective amount of an antibody conjugate composition as described herein to a subject in need thereof, such as a patient that has cancer and is in need of treatment for the cancer. The method includes administering a therapeutically effective amount of an DAC selected from Tables 3a-d. In certain embodiments, the DAC include those with anticancer activity. The DAC 30 selectively delivers an effective dose of an active form of the TPI-VHL protein target degrader moiety to tumor tissue, whereby greater selectivity (i.e., a lower efficacious dose) may be achieved while increasing the therapeutic index (“therapeutic window”) relative to an unconjugated protein target degrader compound. 191 15077.006WO1 It is contemplated that the DAC may be used to treat various hyperproliferative diseases or disorders, e.g. characterized by the overexpression of a tumor antigen. Exemplary hyperproliferative disorders include benign or malignant solid tumors and hematological disorders such as leukemia and lymphoid malignancies. 5 Examples of cancer to be treated herein include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, leukemia or lymphoid malignancies including acute myeloid leukemia, squamous cell cancer, epithelial squamous cell cancer, lung cancer including small- cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung and squamous carcinoma of the lung, cancer of the peritoneum, hepatocellular cancer, gastric or stomach 10 cancer including gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, rectal cancer, colorectal cancer, endometrial or uterine carcinoma, salivary gland carcinoma, kidney or renal cancer, prostate cancer, vulval cancer, thyroid cancer, hepatic carcinoma, anal carcinoma, penile carcinoma, and head and neck cancer. In some embodiments, the DAC may be useful in therapy 15 to treat solid tumors such as lung cancer; non-small cell lung cancer, squamous cell lung cancer, small cell lung cancer, breast cancer, and neuroendocrine cancers such as neuroendocrine prostate cancer, castration-resistant neuroendocrine prostate cancer (NEPC) and lung neuroendocrine tumors. In some embodiments, the DAC may be useful in therapy to treat blood-borne hematological cancers such as leukemias; acute myelogenous leukemia (AML) and 20 myelomas; multiple myeloma (MM). In another aspect, a DAC for use as a medicament is provided. In certain embodiments, the invention provides a DAC for use in a method of treating an individual comprising administering to the individual an effective amount of the antibody conjugate composition in a pharmaceutical composition. In one such embodiment, the method further comprises 25 administering to the individual an effective amount of at least one additional therapeutic agent, e.g., as described herein. In a further aspect, the invention provides for the use of a DAC in the manufacture or preparation of a medicament. In one embodiment, the medicament is for treatment of cancer, the method comprising administering to an individual having cancer an effective amount of the 30 medicament. In one such embodiment, the method further comprises administering to the individual an effective amount of at least one additional therapeutic agent, e.g., as described herein. Dosing can be by any suitable route, e.g. by injections, such as intravenous or subcutaneous injections, depending in part on whether the administration is brief or chronic. 35 Various dosing schedules including but not limited to single or multiple administrations over 192 15077.006WO1 various time-points, bolus administration, and pulse infusion are contemplated herein. The DAC dose can range from about 5 mg/kg (body weight) to about 50 mg/kg, from about 10 µg/kg to about 5 mg/kg, or from about 100 µg/kg to about 1 mg/kg. The DAC dose can be about 100, 200, 300, 400, or 500 µg/kg. The DAC dose can be about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mg/kg. 5 The DAC dose can also be outside of these ranges, depending on the particular conjugate as well as the type and severity of the cancer or disorder being treated. Frequency of administration can range from a single dose to multiple doses per week, or more frequently. In some embodiments, the DAC is administered from about once per month to about five times per week. In some embodiments, the DAC is administered once per week. 10 The DAC can be used either alone or in combination with other therapeutic agents in a therapy regimen. DAC may be administered concurrently in a regimen with one or more other drugs during the same treatment cycle, on the same day of treatment as the one or more other drugs, and, optionally, at the same time as the one or more other drugs. For instance, for cancer therapies given every 3 weeks, the concurrently administered drugs are each administered on 15 day-1 of a 3-week cycle. For instance, a DAC may be co-administered with at least one additional therapeutic agent, such as a chemotherapeutic agent. Such combination therapies encompass combined administration (where two or more therapeutic agents are included in the same or separate formulations), and separate administration, in which case, administration of the DAC can occur prior to, simultaneously, and/or following, administration of the additional 20 therapeutic agent. DAC can also be used in combination with radiation therapy. EXAMPLES General Synthetic Schemes and Examples The following synthetic schemes are provided for purposes of illustration, not limitation. The following examples illustrate the various methods of making compounds described herein. 25 It is understood that one skilled in the art may be able to make these compounds by similar methods or by combining other methods known to one skilled in the art. It is also understood that one skilled in the art would be able to make, in a similar manner as described below by using the appropriate starting materials and modifying the synthetic route as needed. In general, starting materials and reagents can be obtained from commercial vendors or synthesized 30 according to sources known to those skilled in the art of prepared as described herein. Example RV-2 Synthesis of (2S,4R)-1-((S)-17-((4-(benzo[d]thiazol-5-ylamino)-6- (tert-butylsulfonyl)quinolin-7-yl)oxy)-2-(tert-butyl)-4-oxo-6,9,12,15-tetraoxa-3- azaheptadecanoyl)-4-hydroxy-N-((R)-2-(methylamino)-1-(4-(4-methylthiazol-5- yl)phenyl)ethyl)pyrrolidine-2-carboxamide, RV-2 193 15077.006WO1 Step A. Preparation of Int 3a 5 To a mixture of tert-butyl N-[(1R)-1-(4-bromophenyl)-2-hydroxy-ethyl]carbamate (25.0 g, 79.1 mmol, 1.0 eq), 4-methylthiazole (11.8 g, 119 mmol, 1.5 eq), Pd(OAc)₂ (888 mg, 3.95 mmol, 0.05 eq), KOAc (15.5 g, 158 mmol, 2.0 eq) in DMA (250 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 90°C for 12 hrs under N2 atmosphere. The mixture was cooled to 20°C. The residue was diluted with H2O (300 mL) and extracted 10 with EtOAc (300 mL x 2). The combined organic layers were washed with brine (250 mL x 2), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, Petroleum ether/Ethyl acetate=100/1 to 0/1). Compound tert-butyl N-[(1R)-2-hydroxy-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamate Int 3a (25.4 g, 76.0 mmol, 96.1% yield) was obtained as a yellow 15 solid.¹H NMR (400 MHz, DMSO-d6) δ8.98 (s, 1H), 7.45 - 7.42 (m, 2H), 7.40 - 7.36 (m, 2H), 194 15077.006WO1 4.82 (s, 1H), 4.60 - 4.49 (m, 1H), 3.51 (d, J = 5.2 Hz, 3H), 2.46 (s, 3H), 1.37 (s, 9H). LCMS: MS (ESI) m/z 335.2 [M+H] ⁺ Step B. Preparation of Int 3b To a solution of Int 3a (19.5 g, 58.3 mmol, 1.0 eq) in MeCN (200 mL) was added SOCl2 5 (17.3 g, 146 mmol, 10.6 mL, 2.5 eq) in MeCN (200 mL) at -40°C and stirred for 10 min, Py (23.1 g, 292 mmol, 23.5 mL, 5.0 eq) was added and warmed to 25°C. The reaction mixture was stirred for 1 hr at 25°C. The residue was concentrated in vacuum. The crude product was triturated with EtOAc at 25°C for 2 min. The crude product tert-butyl (4R)-4-[4-(4- methylthiazol-5-yl)phenyl]-2-oxo-oxathiazolidine-3-carboxylate Int 3b was used into the next 10 step without further purification. LCMS: MS (ESI) m/z 381.0 [M+H] ⁺ Step C. Preparation of Int 3c To a solution of Int 3b (22.0 g, 57.8 mmol, 1.0 eq) in MeCN (220 mL) and H₂O (220 mL) was added RuCl₃.H₂O (652 mg, 2.89 mmol, 0.05 eq) and NaIO₄ (18.6 g, 86.7 mmol, 4.81 mL, 1.5 eq) at 0°C. The reaction mixture was warmed to 25°C and stirred for 1 hr under N2 15 atmosphere. The residue was poured into ice-water (w/w = 1/1) (300 mL) and stirred for 2 min. The aqueous phase was extracted with DCM (350 mL x 3). The combined organic phase was washed with NaS2SO3 (400mL x 3), brine (400 mL), dried with anhydrous Na2SO4, filtered and concentrated in vacuum. The crude product tert-butyl (4R)-4-[4-(4-methylthiazol-5-yl)phenyl]- 2,2-dioxo-oxathiazolidine-3-carboxylate Int 3c (25 g, crude) was used into the next step without 20 further purification. LCMS: MS (ESI) m/z 397.1 [M+H] ⁺ Step D. Preparation of Int 3d To a solution of 1-(2,4-dimethoxyphenyl)-N-methyl-methanamine (12.1 g, 66.6 mmol, 1.2 eq) in anhydrous MeCN (220 mL) was added Int 3c (22.0 g, 55.5 mmol, 1.0 eq) in anhydrous MeCN (220 mL). The reaction mixture was stirred at 25°C for 1 hr. HCl (1 M) was 25 added and adjusted until pH=~2 at 0°C. The mixture was warmed to 25°C and stirred for 1 hr. The residue was poured into ice-water (w/w = 1/1) (300 mL) and stirred for 2 min. The aqueous phase was extracted with ethyl acetate (350 mL x 3). The combined organic phase was washed with brine (300 mL), dried with anhydrous Na2SO4, filtered and concentrated in vacuum. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm,30 100-200 mesh silica gel, Petroleum ether/Ethyl acetate=100/1, 1/2). Compound tert-butyl N- [(1R)-2-[(2,4-dimethoxyphenyl)methyl-methyl-amino]-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamate Int 3d (12.0 g, 24.1 mmol, 43.5% yield) was obtained as a yellow solid. ¹H NMR (400 MHz, DMSO-d₆) δ 8.98 (s, 1H), 7.44 - 7.36 (m, 4H), 7.02 (d, J = 8.4 Hz, 1H), 6.52 (d, J = 2.4 Hz, 1H), 6.40 (dd, J = 2.0, 8.0 Hz, 1H), 4.72 (s, 1H), 3.75 (s, 3H), 3.73 (s, 195 15077.006WO1 3H), 3.46 (q, J = 13.6 Hz, 2H), 2.62 - 2.53 (m, 2H), 2.46 (s, 3H), 2.13 (s, 3H), 1.38 (s, 9H) LCMS: MS (ESI) m/z 498.1 [M+H] ⁺ Step E. Preparation of Int 3e To a solution of Int 3d (12.0 g, 24.11 mmol, 1.0 eq) in EtOAc (5 mL) was added 5 HCl/EtOAc (4 M, 6.03 mL, 1.0 eq). The reaction mixture was stirred at 25°C for 30 min. The residue was filtered and concentrated in vacuum. The crude product (1R)-N'-[(2,4- dimethoxyphenyl)methyl]-N'-methyl-1-[4-(4-methylthiazol-5-yl)phenyl]ethane-1,2 -diamine Int 3e (16.0 g, crude, HCl) was obtained as a yellow solid. LCMS: MS (ESI) m/z 398.1 [M+H] ⁺ Step F. Preparation of Int 3f 10 To a solution of (2S,4R)-1-[(2S)-2-(tert-butoxycarbonylamino)-3,3-dimethyl-butanoyl]- 4-hydroxy-pyrrolidine-2-carboxylic acid (3.06 g, 8.87 mmol, 1.1 eq), Int 3e (3.5 g, 8.06 mmol, 1.0 eq, HCl) in DMF (35 mL) were added HOBt(1.63 g, 12.1 mmol, 1.5 eq), DIEA (5.21 g, 40.3 mmol, 7.02 mL, 5.0 eq), and EDCI (2.32 g, 12.1 mmol, 1.5 eq). The reaction mixture was stirred at 25°C for 1 hr. The mixture was cooled to 0°C. The residue was poured into ice-water 15 (w/w = 1/1) (35 mL) and stirred for 2 min. The aqueous phase was extracted with ethyl acetate (30 mL x 3). The combined organic phase was washed with brine (35 mL x 2), dried with anhydrous Na2SO4, filtered and concentrated in vacuum. The residue was purified by column (SiO2, Petroleum ether/Ethyl acetate = 0:1). Compound tert-butyl N-[(1S)-1-[(2S,4R)-2-[[(1R)- 2-[(2,4-dimethoxyphenyl)methyl-methyl-amino]-1-[4-(4-methylthiazol-5- 20 yl)phenyl]ethyl]carbamoyl]-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]carbamate Int 3f (3.22 g, 4.45 mmol, 55.2% yield) was obtained as a yellow solid. ¹H NMR: (400 MHz, DMSO-d₆) δ 8.98 (s, 1H), 7.42 - 7.33 (m, 4H), 7.01 (d, J = 8.4 Hz, 1H), 6.49 (d, J = 2.4 Hz, 1H), 6.39 (dd, J = 2.4, 8.4 Hz, 1H), 5.11 (d, J = 3.6 Hz, 1H), 4.95 (q, J = 7.6 Hz, 1H), 4.45 (t, J = 7.6 Hz, 1H), 4.28 (s, 1H), 4.14 (d, J = 9.2 Hz, 1H), 3.72 (d, J = 1.6 Hz, 6H), 3.63 - 3.52 (m, 2H), 25 3.44 (s, 2H), 2.60 (t, J = 6.8 Hz, 2H), 2.46 (s, 3H), 2.16 (s, 3H), 2.05 - 2.00 (m, 1H), 1.79 - 1.69 (m, 1H), 1.38 (s, 9H), 0.93 (s, 9H) LCMS: MS (ESI) m/z 724.3 [M+H] ⁺ Step G. Preparation of Int 3g 196 15077.006WO1 To a solution of Int 3f (0.2 g, 276 μmol, 1 eq) in EtOAc (2 mL) was added HCl/EtOAc (4 M, 6 mL). The reaction mixture was stirred at 20°C for 1 hr. The mixture was concentrated in vacuum. Afford (2S,4R)-1-[(2S)-2-amino-3,3-dimethyl-butanoyl]-N-[(1R)-2-[(2,4- dimethoxyphenyl)methyl-methyl-amino]-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]-4-hydroxy- 5 pyrrolidine-2-carboxamide Int 3g (170 mg, 257 μmol, 93.19% yield, HCl) as white solid. LCMS: MS (ESI) m/z 624.2 [M+H]⁺ Step H. Preparation of Int 3h 197 15077.006WO1 A solution of 6-bromo-4-chloro-7-methoxy-quinoline (8 g, 29.4 mmol, 1 eq), Na2CO3 (7.78 g, 73.4 mmol, 2.5 eq) and Pd(PPh₃)₄ (1.7 g, 1.47 mmol, 0.05 eq) in DMF (80 mL) was 5 deoxygenated in a closed tube for 10 minutes. 2- methylpropane-2-thiol (2.65 g, 29.3 mmol, 3.31 mL, 1 eq) was added. The reaction mixture was heated to 140°C for 12 hrs. The reaction mixture was partitioned between EtOAc and a saturated solution of sodium thiosulfate and sodium bicarbonate (v/v 5:1). The aqueous layer was extracted twice with EtOAc (80 mL x 2) and the combined EtOAc layers were washed with brine (40 mL x 2), dried over sodium sulfate 10 and concentrated. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0~45% Ethyl acetate/Petroleum ethergradient @ 80 mL/min). Compound 6-tert-butylsulfanyl-4-chloro-7-methoxy-quinoline Int 3h (6.6 g, 23.4 mmol, 79.78% yield) was obtained as a white solid. LCMS: MS (ESI) m/z 282.1 [M+H]⁺ Step I. Preparation of Int 3i 15 1,3-benzothiazol-5-amine (266 mg, 1.77 mmol, 1 eq) and Int 3h (500 mg, 1.77 mmol, 1 eq) were taken up into a microwave tube in propan-2-ol (10 mL). The sealed tube was heated at 198 15077.006WO1 150°C for 2 hrs under microwave. The reaction mixture was concentrated in vacuum. The crude product N-(6-tert-butylsulfanyl-7-methoxy-4-quinolyl)-1,3-benzothiazol-5-amine Int 3i (0.76 g, crude) was obtained as yellow solid and used into the next step without further purification. LCMS: MS (ESI) m/z 396.0 [M+H]⁺ 5 Step J. Preparation of Int 3j To a solution of Int 3i (2.1 g, 5.31 mmol, 1 eq) in MeOH (20 mL) and H₂O (20 mL) was added Oxone (4.9 g, 7.96 mmol, 1.5 eq). The reaction mixture was stirred at 25°C for 1 hr. The mixture was quenched with Na2CO3 until pH=~8, and extracted with DCM/i-PrOH=3/1 (30 mL x 3). The combined organic layers were dried over Na₂SO₄, filtered and concentrated under 10 reduced pressure to give a residue. The crude product was triturated with MTBE (50 mL) at 25°C for 30 min. Compound N-(6-tert-butylsulfonyl-7-methoxy-4-quinolyl)-1,3-benzothiazol-5- amine Int 3j (1.9 g, 4.44 mmol, 83.70% yield) was obtained as a yellow solid. LCMS: MS (ESI) m/z 428.0 [M+H]⁺ Step K. Preparation of Int 3k 15 To a solution of Int 3j (2.6 g, 6.08 mmol, 1 eq) in DMF (30 mL) was added isopropylsulfanyl sodium (2.98 g, 30.4 mmol, 5 eq). The reaction was then heated to 145 °C for 1 hr. The mixture was quenched with TFA until pH=~6. The reaction mixture was filtered and the filter cake was washed with 30 mL of H2O, dried in vacuum to give or afford product. The crude product 4-(1,3-benzothiazol-5-ylamino)-6-tert-butylsulfonyl-quinolin-7-ol Int 3k (1.4 g, 20 3.39 mmol, 55.67% yield) was obtained as brown solid and used into the next step without further purification. LCMS: MS (ESI) m/z 414.2 [M+H]⁺ Step L. Preparation of Int 3l To a solution of Int 3k (834 mg, 2.02 mmol, 1 eq) and tert-butyl 2-[2-[2-[2-[2-(p- tolylsulfonyloxy)ethoxy]ethoxy]ethoxy]ethoxy]acetate (1.4 g, 3.03 mmol, 1.5 eq) in NMP (20 25 mL) was added K2CO3 (836 mg, 6.05 mmol, 3 eq). The reaction mixture was stirred at 90°C for 12 hrs. The reaction mixture was quenched by addition H₂O 20 mL, and then extracted with DCM:i-PrOH=3:1(30 mL x 2). The combined organic layers were washed with brine (10 mL x 2). dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash30 Column, Eluent of 0~10% DCM/MeOH @ 40 mL/min). The crude product tert-butyl 2-[2-[2- [2-[2-[[4-(1,3-benzothiazol-5-ylamino)-6-tert-butylsulfonyl-7- quinolyl]oxy]ethoxy]ethoxy]ethoxy]ethoxy]acetate Int 3l (350 mg, 497 μmol, 24.64% yield) was obtained as brown oil and used into the next step without further purification. LCMS: MS (ESI) m/z 704.3 [M+H]⁺ 35 Step M. Preparation of Int 3m 199 15077.006WO1 To a solution of Int 3l (300 mg, 426 μmol, 1 eq) in DCM (3 mL) was added TFA (1.5 mL). The reaction mixture was stirred at 25°C for 1 hr. The reaction mixture was concentrated in vacuum. The crude product 2-[2-[2-[2-[2-[[4-(1,3-benzothiazol-5-ylamino)-6-tert- butylsulfonyl-7-quinolyl]oxy]ethoxy]ethoxy]ethoxy]ethoxy]acetic acid Int 3m (250 mg, crude) 5 was obtained as brown oil and used into the next step without further purification. LCMS: MS (ESI) m/z 648.0 [M+H]⁺ Step N. Preparation of Int 3n To a solution of Int 3g (250 mg, 379 μmol, 1 eq, HCl) in DMF (3 mL) were added Int 3m (221 mg, 341 μmol, 0.9 eq), DIEA (245 mg, 1.89 mmol, 330 μL, 5 eq) and HATU (216 mg,10 568 μmol, 1.5 eq). The mixture was stirred at 20°C for 1 hr. The residue was poured into ice- water (w/w = 1/1) (5 mL). The aqueous phase was extracted with DCM/i-PrOH=3/1 (5 mL x 2). The combined organic phase was dried with anhydrous Na₂SO₄, filtered and concentrated in vacuum. The residue was purified by silica gel chromatography (column height: 100 mm, diameter: 60 mm, 100-200 mesh silica gel, MeOH/Ethyl acetate=0/1, 1/1) to afford (2S,4R)-1-15 [(2S)-2-[[2-[2-[2-[2-[2-[[4-(1,3-benzothiazol-5-ylamino)-6-tert-butylsulfonyl-7- quinolyl]oxy]ethoxy]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-N-[(1R)-2- [(2,4-dimethoxyphenyl)methyl-methyl-amino]-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]-4- hydroxy-pyrrolidine-2-carboxamide Int 3n (250 mg, 199 μmol, 52.67% yield) as yellow solid. LCMS: MS (ESI) m/z 1253.2 [M+H]⁺ 20 Step O. Preparation of RV-2 A mixture of Int 3n (40 mg, 31.9 μmol, 1 eq) in TFA (2 mL) was stirred at 110°C for 9 hrs. The mixture was concentrated in vacuum, the residue was dissolved in (2 mL), K₂CO₃ (22.1 mg, 160 μmol, 5 eq) was added. The reaction mixture was stirred at 20°C for 1 hr. The mixture was filtered and the filtrate was concentrated in vacuum. The residue was purified by25 prep-HPLC (column: Phenomenex Gemini-NX 80*40mm*3um;mobile phase: [H2O(0.1%TFA)- ACN];gradient:10%-40% B over 20.0 min) to give RV-2 (3.5 mg, 2.82 μmol, 8.83% yield, 98% purity, TFA) as yellow solid.¹H NMR: (400 MHz, DMSO-d₆) δ 11.55 (s, 1H), 9.54 (s, 1H), 9.26 (s, 1H), 9.02 (s, 1H), 8.87 (d, J = 8.8 Hz, 1H), 8.48 (d, J = 7.2 Hz, 2H), 8.39 (d, J = 8.4 Hz, 2H), 8.22 (d, J = 2.0 Hz, 1H), 7.60 (dd, J = 2.0, 8.4 Hz, 1H), 7.57-7.45 (m, 5H), 7.39 (d, J = 9.2 Hz, 30 1H), 6.83 (d, J = 7.2 Hz, 1H), 5.38-5.24 (m, 1H), 4.55 (d, J = 9.2 Hz, 1H), 4.46-4.33 (m, 4H), 4.03-3.96 (m, 2H), 3.88 (d, J = 4.0 Hz, 2H), 3.73-3.52 (m, 16H), 2.68 (s, 3H), 2.46 (s, 3H), 2.07 (dd, J = 7.2, 12.4 Hz, 1H), 1.89-1.78 (m, 1H), 1.36 (s, 9H), 1.01-0.93 (m, 9H). LCMS: MS (ESI) m/z 1103.4 [M+H]⁺ Example KV-13 Synthesis of (2S,4R)-1-((S)-2-(4-(4-((S)-4-(4-(5-((S)-2-35 amino-3-cyano-4-methyl-4,5,6,7-tetrahydrobenzo[b]thiophen-4-yl)-1,2,4-oxadiazol-3- 200 15077.006WO1 yl)pyrimidin-2-yl)-3-methyl-1,4-diazepan-1-yl)butoxy)-1H-1,2,3-triazol-1-yl)-3- methylbutanoyl)-4-hydroxy-N-((R)-2-(methylamino)-1-(4-(4-methylthiazol-5- yl)phenyl)ethyl)pyrrolidine-2-carboxamide, KV-13 Step A. Preparation of Int 4a 5 To a mixture of tert-butyl N-[(2R)-2-[[(2S,4R)-1-[(2S)-2-azido-3-methyl-butanoyl]-4- hydrox-pyrrolidine-2-carbonyl]amino]-2-[4-(4-methylthiazol-5-yl)phenyl]ethyl]-N-methyl- 10 carbamate (0.15 g, 256 μmol, 1.0 eq) and tert-butyl-(4-ethynoxybutoxy)-dimethyl-silane (175 mg, 768 μmol, 3.0 eq) in THF (1 mL), t-BuOH (1 mL) and H₂O (1 mL) were added sodium;(2R)-2-[(1S)-1,2-dihydroxyethyl]-4-hydroxy-5-oxo-2H-furan-3-olate (76.1 mg, 384 201 15077.006WO1 μmol, 1.5 eq) and copper;sulfate (20.4 mg, 128. μmol, 19.7 μL, 0.5 eq) in one portion at 20°C. Then the mixture was allowed to warm to 30°C and stirred for 0.5 hr. The mixture was diluted with water (20 mL) and extracted with EtOAc (15 mL x 3). The organic layer was washed with brine, dried over Na2SO4, filtered and concentrated. Compound tert-butyl N-[(2R)-2-[[(2S,4R)- 5 1-[(2S)-2-[4-[4-[tert-butyl(dimethyl)silyl]oxybutoxy]triazol-1-yl]-3-methyl-butanoyl]-4- hydroxy-pyrrolidine-2-carbonyl]amino]-2-[4-(4-methylthiazol-5-yl) phenyl]ethyl]-N-methyl- carbamate Int 4a (0.28 g, crude) was obtained as yellow oil. LCMS: MS (ESI) m/z 814.4 [M+H]⁺ Step B. Preparation of Int 4b 10 To a mixture of Int 4a (0.28 g, 343 μmol, 1.0 eq) in DCM (4 mL) were added Ac2O (45.6 mg, 447 μmol, 41.9 μL, 1.3 eq), Et3N (69.6 mg, 688 μmol, 95.7 μL, 2.0 eq) and DMAP (4.20 mg, 34.4 μmol, 0.1 eq) in one portion at 20°C and stirred for 0.5 hr. The mixture was diluted with water (20 mL) and extracted with EtOAc (15 mL x 3). The organic layer was washed with brine, dried over Na2SO4, filtered and concentrated. Compound [(3R,5S)-5-15 [[(1R)-2-[tert-butoxycarbonyl(methyl)amino]-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]-1-[(2S)-2-[4-[4-[tert-butyl(dimethyl) silyl]oxybutoxy]triazol-1-yl]- 3-methyl-butanoyl]pyrrolidin-3-yl] acetate Int 4b (0.29 g, crude) was obtained as yellow oil. LCMS: MS (ESI) m/z 856.4 [M+H]⁺ Step C. Preparation of Int 4c 20 To a mixture of Int 4b (0.29 g, 339 μmol, 1.0 eq) in MeOH (3 mL) was added acetyl chloride (39.9 mg, 508 μmol, 36.1 μL, 1.5 eq) in one portion at 20°C and stirred for 0.5 hr. The mixture was diluted with water (20 mL). Then the aqueous phase was extracted with ethyl acetate (10 mL x 3). The combined organic phase was washed with brine (10 mL x 2), dried with anhydrous Na2SO4, filtered and concentrated in vacuum. Compound [(3R,5S)-5-[[(1R)-2-25 [tert-butoxycarbonyl(methyl) amino]-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]-1- [(2S)-2-[4-(4-hydroxybutoxy)triazol-1-yl]-3-methyl-butanoyl]pyrrolidin-3-yl] acetate Int 4c (0.25 g, crude) was obtained as yellow oil. LCMS: MS (ESI) m/z 742.4 [M+H]⁺ Step D. Preparation of Int 4d To a mixture of Int 4c (0.22 g, 297 μmol, 1.0 eq) in DCM (4 mL) was added Dess- 30 Martin reagent (201.2 mg, 474 μmol, 147.00 μL, 1.6 eq) in one portion at 20°C and stirred at 20°C for 0.5 hr. The mixture was filtered and concentrated. Then the residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, Ethyl acetate/MeOH=1/0, 10/1) to afford [(3R,5S)-5-[[(1R)-2-[tert- butoxycarbonyl(methyl)amino]-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]-1-[(2S)-3- 202 15077.006WO1 methyl-2-[4-(4-oxobutoxy)triazol-1-yl]butanoyl]pyrrolidin-3-yl] acetate Int 4d (0.14 g, 189.22 μmol, 63.81% yield) as yellow solid. LCMS: MS (ESI) m/z 740.3 [M+H]⁺ Step E. Preparation of Int 4e To a mixture of Int 4d (0.12 g, 162. μmol, 1.0 eq) and (4S)-2-amino-4-methyl-4-[3-[2- 5 [(2S)-2-methyl-1,4-diazepan-1-yl]pyrimidin-4-yl]-1,2,4-oxadiazol-5-yl]-6,7-dihydro-5H- benzothiophene-3-carbonitrile (65.8 mg, 146 μmol, 0.9 eq) in MeOH (3 mL) was added NaBH3CN (40.8 mg, 649 μmol, 4.0 eq) in one portion at 20°C and stirred for 2 hrs. The residue was poured into water (20 mL) and stirred for 2 min. The aqueous phase was extracted with ethyl acetate (10 mL x 3). The combined organic phase was washed with brine (10 mL x 2), 10 dried with anhydrous Na₂SO₄, filtered and concentrated in vacuum. The residue was purified by column chromatography (SiO2, Ethyl acetate/MeOH=1/0 to 3/1). Compound [(3R,5S)-1-[(2S)- 2-[4-[4-[(3S)-4-[4-[5-[(4S)-2-amino-3-cyano-4-methyl-6,7-dihydro-5H-benzothiophen-4-yl]- 1,2,4-oxadiazol-3-yl] pyrimidin-2-yl]-3-methyl-1,4-diazepan-1-yl]butoxy]triazol-1-yl]-3- methyl-butanoyl]-5-[[(1R)-2-[tert-butoxycarbonyl(methyl)amino]-1-[4-(4-methylthiazol-5- 15 yl)phenyl]ethyl] carbamoyl]pyrrolidin-3-yl] acetate Int 4e (0.07 g, 59.60 μmol, 36.75% yield) was obtained as a yellow solid. LCMS: MS (ESI) m/z 1174.5 [M+H]⁺ Step F. Preparation of Compound KV-3 To a mixture of Int 4e (0.05 g, 44.2 μmol, 1.0 eq) in EtOAc (1 mL) was added HCl/EtOAc (1.5 mL) in one portion at 20°C and stirred at 20°C for 0.5 hr. The mixture 20 waspurified by prep-HPLC(column: 3_Phenomenex Luna C1875*30mm*3um;mobile phase: [H2O (0.1%TFA)-ACN];gradient:15%-45% B over 8.0 min) to give KV-13 (65 mg, crude, HCl) as yellow solid.¹H NMR: (400 MHz, DMSO-d₆) δ = 9.03-8.98 (m, 1H), 8.65 (d, J = 4.0 Hz, 1H), 7.74-7.66 (m, 1H), 7.55-7.46 (m, 4H), 7.26 (dd, J = 4.4, 7.6 Hz, 1H), 5.40-5.25 (m, 1H), 5.21-5.09 (m, 1H), 4.98-4.78 (m, 1H), 4.48-4.30 (m, 3H), 4.17-4.06 (m, 2H), 3.82 (d, J = 7.2 Hz, 25 2H), 3.72-3.65 (m, 2H), 3.59-3.54 (m, 2H), 3.43-3.35 (m, 4H), 3.32-3.19 (m, 4H), 2.68 (s, 3H), 2.46 (s, 3H), 2.13-2.03 (m, 2H), 1.98-1.81 (m, 8H), 1.78 (s, 3H), 1.75-1.69 (m, 2H), 1.22 (s, 3H), 1.14 (d, J = 4.0 Hz, 3H), 1.03 (d, J = 6.4 Hz, 3H), 0.69 (d, J = 6.4 Hz, 3H). LCMS: MS (ESI) m/z 1032.5 [M+H]⁺ The following compounds was prepared similar to KV-13 using the appropriate 30 compound as starting material. 203 15077.006WO1 Example KV-14 Synthesis of (2S,4R)-1-((S)-2-(4-(4-((S)-4-(4-(5-((S)-2-amino-3- cyano-4-methyl-4,5,6,7-tetrahydrobenzo[b]thiophen-4-yl)-1,2,4-oxadiazol-3-yl)pyrimidin-2-yl)- 3-methyl-1,4-diazepan-1-yl)butoxy)-1H-1,2,3-triazol-1-yl)-3-methylbutanoyl)-N-((S)-1-(5- 5 fluoro-2-hydroxy-4-(4-methylthiazol-5-yl)phenyl)ethyl)-4-hydroxypyrrolidine-2-carboxamide, KV-14 204 15077.006WO1 Step A. Preparation of Int 5a. To a solution of 1-(4-bromo-5-fluoro-2-hydroxy-phenyl) ethanone (20 g, 85.82 mmol, 1 eq) in DMF (300 mL) were added BnBr (17.6 g, 103 mmol, 12.2 mL, 1.2 eq) and K₂CO₃ (23.7 5 g, 172 mmol, 2 eq) at 20°C. The reaction mixture was heated to 80 °C and stirred for 16 hr. The reaction mixture was quenched by addition water 300 mL, and extracted with EtOAc 200 mL x3, the combined organic layers were washed with brine 300 mL, dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO₂, Petroleum ether/Ethyl acetate=1/0 to 10/1). Compound 1-(2-benzyloxy- 10 4-bromo-5-fluoro-phenyl) ethenone Int 5a (32.0 g, crude) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d₆) δ7.67 (d, J = 5.6 Hz, 1H), 7.52 (t, J = 8.4 Hz, 3H), 7.47-7.40 (m, 2H), 7.38-7.35 (m, 1H), 5.27 (s, 2H), 2.50 (s, 3H). Step B. Preparation of Int 5b To a solution of Int 5a (15.0 g, 46.4 mmol, 1 eq) in THF (200 mL) were added 15 tetraethoxytitanium (31.8 g, 139 mmol, 28.9 mL, 3 eq) and (S)-2-methylpropane-2-sulfinamide (6.75 g, 55.7 mmol, 1.2 eq) at 20°C. Then the reaction mixture was heated to 80°C and stirred for 16 hr. The reaction mixture was quenched by addition water 200 mL, and then extracted with EtOAc 200 ml x3. The combined organic layers were washed with brine 400, dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was 20 purified by column chromatography (SiO2, Petroleum ether/Ethyl acetate=1/0 to 10/1). Compound (NE,S)-N-[1-(2-benzyloxy-4-bromo-5-fluoro-phenyl)ethylidene]-2-methyl-propane- 2-sulfinamide Int 5b (34.0 g, crude) was obtained as a white solid.¹H NMR: (400 MHz, DMSO-d6) δ = 7.60 (d, J = 5.6 Hz, 1H), 7.44-7.35 (m, 6H), 5.21 (s, 2H), 2.56 (s, 3H), 1.13 (s, 9H). LCMS: MS (ESI) m/z 426.0 [M+H]⁺ 25 Step C. Preparation of Int 5c To a mixture of Int 5b (15.0 g, 35.2 mmol, 1 eq) ,trichlorocerium (4.34 g, 17.6 mmol, 1.11 mL, 0.5 eq) in THF (200 mL) was added NaBH4 (2.66 g, 70.4 mmol, 2 eq) at -65°C. The 205 15077.006WO1 reaction mixture was slowly warmed to 20°C for 16 hr. The reaction mixture was quenched by addition ice water 400 mL, and then extracted with EtOAc (400 mL*3). The combined organic layers were washed with brine 400 mL, dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, 5 Petroleum ether/Ethyl acetate=1/0 to 3/1). Compound (S)-N-[(1S)-1-(2-benzyloxy-4-bromo-5- fluoro-phenyl)ethyl]-2-methyl-propane-2-sulfinamide Int 5c (18 g, crude) was obtained as white oil.¹H NMR: (400 MHz, DMSO-d6) δ7.47-7.35 (m, 7H), 5.76-5.73 (m, 1H), 5.16 (d, J = 2.5 Hz, 2H), 4.67-4.63 (m, 1H), 1.30 (d, J = 6.8 Hz, 3H), 1.08 (s, 9H). LCMS: MS (ESI) m/z 428.1 [M+H]⁺ 10 Step D. Preparation of Int 5d To a solution of Int 5c (17.5 g, 40.9 mmol, 1 eq) in DMA (250 mL) were added 4- methylthiazole (12.2 g, 123 mmol, 11.2 mL, 3 eq), KOAc (8.02 g, 81.7 mmol, 2 eq) and Pd(OAc)₂ (917 mg, 4.09 mmol, 0.1 eq) at 20°C. Then the reaction mixture was heated to 130°C and stirred for 16hr. The reaction mixture was quenched by addition water 600 mL, and then 15 extracted with EtOAC 200 mL x3. The combined organic layers were washed with brine 400 mL, dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, Petroleum ether/Ethyl acetate=1/0 to 1/1). Compound (S)-N-[(1S)-1-[2-benzyloxy-5-fluoro-4-(4-methylthiazol-5-yl) phenyl] ethyl]- 2-methyl-propane-2-sulfinamide Int 5d (10.5 g, 23.5 mmol, 57.55% yield) was obtained as a 20 yellow solid.¹H NMR: (400 MHz, DMSO-d6) δ9.10 (s, 1H), 7.49-7.44 (m, 2H), 7.44-7.40 (m, 2H), 7.37-7.32 (m, 2H), 7.09 (d, J = 6.0 Hz, 1H), 5.78-5.74 (m, 1H), 5.20 (s, 2H), 4.79-4.70 (m, 1H), 2.29 (s, 3H), 1.37 (d, J = 6.8 Hz, 3H), 1.11 (s, 9H). LCMS: MS (ESI) m/z 447.1 [M+H]⁺ Step E. Preparation of Int 5e To a solution of Int 5d (10.0 g, 22.4 mmol, 1 eq) in EtOAc (20 mL) was added 25 HCl/EtOAc (4 M, 224 mL, 40 eq). The reaction mixture was stirred at 25°C for 1hr. The reaction mixture was concentrated under reduced pressure. The crude product was triturated with MTBE at 20^oC for 60 min. Compound (1S)-1-[2-benzyloxy-5-fluoro-4-(4-methylthiazol-5- yl) phenyl] ethanamine Int 5e (10.0 g, crude, 2HCl) was obtained as a yellow solid.¹H NMR: (400 MHz, DMSO-d6) δ9.15 (s, 1H), 8.57 (s, 2H), 8.29-8.28 (m, 1H), 7.62-7.58 (m, 1H), 7.51- 30 7.47 (m, 2H), 7.42 (m, 3H), 7.21 (d, J = 6.4 Hz, 1H), 5.24 (m, 2H), 4.13-3.96 (m, 1H), 2.27 (s, 3H), 1.51 (d, J = 6.8 Hz, 3H). LCMS: MS (ESI) m/z 343.1 [M+H]⁺ Step F. Preparation of Int 5f To a solution of Int 5e (9.00 g, 26.3 mmol, 1 eq) in DMF (100 mL) were added DIEA (13.6 g, 105 mmol, 18.3 mL, 4 eq), (2S,4R)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-methyl- 35 butanoyl]-4-hydroxy-pyrrolidine-2-carboxylic acid (8.68 g, 26.28 mmol, 1 eq), EDCI (5.04 g, 206 15077.006WO1 26.28 mmol, 1 eq) and HOBt (5.33 g, 39.4 mmol, 1.5 eq) at 25°C. Then the reaction mixture was stirred at 25°C for 2 hr. The reaction mixture was quenched by addition water 300 mL, and then extracted with EtOAc 100 ml x3. The combined organic layers were washed with brine 200 mL, dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. 5 The residue was purified by column chromatography (SiO2, Petroleum ether/Ethyl acetate=1/0 to 0/1). Compound tert-butyl N-[(1S)-1-[(2S,4R)-2-[[(1S)-1-[2-benzyloxy-5-fluoro-4-(4- methylthiazol-5-yl) phenyl] ethyl] carbamoyl]-4-hydroxy-pyrrolidine-1-carbonyl]-2-methyl- propyl] carbamate Int 5f (15.0 g, 22.9 mmol, 87.16% yield) was obtained as a yellow solid.¹H NMR: (400 MHz, DMSO-d6) δ9.09 (s, 1H), 8.34 (d, J = 7.6 Hz, 1H), 7.50-7.45 (m, 2H), 7.40 (t, 10 J = 7.4 Hz, 2H), 7.34 (d, J = 7.2 Hz, 1H), 7.19 (m, 1H), 7.08 (d, J = 6.0 Hz, 1H), 6.69 (d, J = 8.4 Hz, 1H), 5.20 (s, 2H), 5.11 (d, J = 3.3 Hz, 1H), 4.43 (t, J = 8.0 Hz, 1H), 4.32-4.29 (m, 1H), 4.04- 4.01 (m, 1H), 3.58 (s, 1H), 2.28 (s, 3H), 2.09-2.01 (m, 1H), 1.96-1.90 (m, 1H), 1.83- 1.73(m, 1H), 1.38 (s, 9H), 1.34 (m, 3H), 0.90-0.83 (m, 6H). LCMS: MS (ESI) m/z 655.1 [M+H]⁺ Step G. Preparation of Int 5g 15 To a solution of Int 5f (7.00 g, 10.7 mmol, 1 eq) in DCM (70 mL) was added BBr3 (2 M, 16.0 mL, 3 eq) at -78°C. Then the reaction mixture was slowly warmed to 20°C and stirred for 12hr. The reaction mixture was quenched by addition water 100 mL. The pH of the mixture was adjusted to 8 with NaHCO3(aq), and extracted with( DCM :iPrOH=3：1) 50 mL x3. The combined organic layers were dried over brine, filtered and concentrated under reduced pressure20 to give a residue. Compound (2S,4R)-1-[(2S)-2-amino-3-methyl-butanoyl]-N-[(1S)-1-[5-fluoro- 2-hydroxy-4-(4-methylthiazol-5-yl) phenyl] ethyl]-4-hydroxy-pyrrolidine-2-carboxamide Int 5g (4.80 g, crude) was obtained as a yellow solid.¹H NMR: (400 MHz, DMSO-d6) δ9.08 (s, 1H), 8.28 (d, J = 8.0 Hz, 1H), 7.07 (m, 1H), 6.89-6.75 (m, 1H), 5.15-5.02 (m, 2H), 4.46-4.42 (m, 1H), 4.34-4.17 (m, 1H), 3.52-3.49 (m, 2H), 3.21 (d, J = 5.6 Hz, 1H), 2.33 (s, 3H), 2.07-2.02 (m, 1H), 25 1.92-1.71 (m, 3H), 1.30 (d, J = 6.8 Hz, 3H), 0.91-0.87 (m, 3H), 0.79 (d, J = 6.4 Hz, 3H). LCMS: MS (ESI) m/z 465.1 [M+H]⁺ Step H. Preparation of Int 5h To a solution of Int 5g (1.30 g, 2.59 mmol, 1 eq, HCl) in CH₃CN (15 mL) were added 2- azido-1,3-dimethyl-4,5-dihydroimidazol-1-ium;hexafluorophosphate (888 mg, 3.11 mmol, 1.2 30 eq), DIEA (1.01 g, 7.78 mmol, 1.36 mL, 3 eq) at 0°C. The reaction mixture was stirred at 0°C for 1hr. The reaction mixture was quenched by addition water 50 mL, and then extracted with EtOAc 20x3 ml. The combined organic layers were washed with brine 40x3 mL, dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, Petroleum ether/Ethyl acetate=1/0 to 0/1). 35 Compound (2S,4R)-1-[(2S)-2-azido-3-methyl-butanoyl]-N-[(1S)-1-[5-fluoro-2-hydroxy-4-(4- 207 15077.006WO1 methylthiazol-5-yl)phenyl]ethyl]-4-hydroxy-pyrrolidine-2-carboxamide Int 5h (0.70 g, 1.43 mmol, 54.99% yield) was obtained as a brown solid. LCMS: MS (ESI) m/z 491.1 [M+H]⁺ Step I. Preparation of Int 5i To a mixture of Int 5h (300 mg, 612 μmol, 1.0 eq) and tert-butyl-(4-ethynoxybutoxy)- 5 dimethyl-silane (182 mg, 795 μmol, 1.3 eq) in THF (2 mL), t-BuOH (2 mL) and H2O (2 mL) were added sodium;(2R)-2-[(1S)-1,2-dihydroxyethyl]-4-hydroxy-5-oxo-2H-furan-3-olate (182 mg, 917 μmol, 1.5 eq) and copper;sulfate (48.8 mg, 306 μmol, 46.9 μL, 0.5 eq) in one portion at 20°C. Then the mixture was allowed to warm to 30°C and stirred for 2 hrs. The mixture was diluted with water (30 mL) and extracted with EtOAc (20 mL x 3). The organic layer was 10 washed with brine, dried over Na₂SO₄, filtered and concentrated. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, Ethyl acetate/MeOH=1/0, 10/1) to afford (2S,4R)-1-[(2S)-2-[4-[4-[tert-butyl(dimethyl) silyl]oxybutoxy]triazol-1-yl]-3-methyl-butanoyl]-N-[(1S)-1-[5-fluoro-2-hydroxy-4-(4- methylthiazol-5-yl)phenyl]ethyl]-4-hydroxy-pyrrolidine-2-carboxamide Int 5i (0.17 g, 236.46 15 μmol, 38.66% yield) as yellow solid.¹H NMR: (400 MHz, MeOD-d4) δ8.97 (s, 1H), 7.67-7.48 (m, 1H), 7.14-6.89 (m, 1H), 6.79 (d, J = 6.4 Hz, 1H), 5.26 (q, J = 7.2 Hz, 1H), 5.15 (d, J = 10.4 Hz, 1H), 4.54 (t, J = 8.4 Hz, 1H), 4.48 (s, 1H), 4.18-4.11 (m, 2H), 3.96-3.79 (m, 2H), 3.71 (t, J = 6.0 Hz, 2H), 2.60-2.47 (m, 1H), 2.38 (s, 3H), 2.23 (dd, J = 8.0, 13.2 Hz, 1H), 2.06-1.94 (m, 1H), 1.89-1.77 (m, 2H), 1.73-1.63 (m, 2H), 1.50 (d, J = 7.2 Hz, 3H), 1.15-1.09 (m, 3H), 0.91 (s, 9H), 20 0.78 (d, J = 6.4 Hz, 3H), 0.07 (s, 6H). LCMS: MS (ESI) m/z 719.3 [M+H]⁺ Step J. Preparation of Int 5j To a mixture of Int 5i (0.17 g, 236 μmol, 1.0 eq) in DCM (5 mL) were added DMAP (2.89 mg, 23.7 μmol, 0.1 eq), Ac₂O (62.8 mg, 615 μmol, 57.74 μL, 2.6 eq) and Et₃N (47.9 mg, 473 μmol, 65.8 μL, 2.0 eq) in one portion at 20°C and stirred for 2 hrs. The mixture was filtered 25 and concentrated. Then the residue was diluted with water and extracted with DCM (20 mL x 3). The organic layer was washed with brine, dried over Na₂SO₄, filtered and concentrated. Compound [2-[(1S)-1-[[(2S,4R)-4-acetoxy-1-[(2S)-2-[4-[4-[tert- butyl(dimethyl)silyl]oxybutoxy]triazol-1-yl]-3-methyl-butanoyl]pyrrolidine-2- carbonyl]amino]ethyl]-4-fluoro-5-(4-methylthiazol-5-yl)phenyl] acetate Int 5j (0.24 g, crude) 30 was obtained as yellow oil. LCMS: MS (ESI) m/z 803.3 [M+H]⁺ Step K. Preparation of Int 5k 208 15077.006WO1 To a mixture of Int 5j (0.24 g, 299 μmol, 1.0 eq) in MeOH (4 mL) was added acetyl chloride (35.2 mg, 448 μmol, 31.9 μL, 1.5 eq) in one portion at 20°C, and then stirred at 20°C 5 for 2 hrs. The residue was poured into ice-water (30 mL) and stirred for 2 min. The aqueous phase was extracted with ethyl acetate (20 mL x 3). The combined organic phase was washed with brine (20 mL x 2), dried with anhydrous Na2SO4, filtered and concentrated in vacuum. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, Ethyl acetate/MeOH=1/0, 10/1) to afford [2-[(1S)-1-[[(2S,4R)-4-10 acetoxy-1-[(2S)-2-[4-(4-hydroxybutoxy)triazol-1-yl]-3-methyl-butanoyl]pyrrolidine-2- 209 15077.006WO1 carbonyl]amino]ethyl]-4-fluoro-5-(4-methylthiazol-5-yl) phenyl] acetate Int 5k (0.13 g, 188.74 μmol, 63.15% yield) as yellow solid. LCMS: MS (ESI) m/z 689.3 [M+H]⁺ Step L. Preparation of Int 5l To a mixture of Int 5k (0.12 g, 174 μmol, 1.0 eq) in DCM (3 mL) was added Dess- 5 Martin (118 mg, 279 μmol, 86.4 μL, 1.6 eq) in one portion at 20°C and stirred for 1 hr. The mixture was diluted with water (20 mL) and extracted with EtOAc (15 mL x 3). The organic layer was washed with brine, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, Ethyl acetate/MeOH=1/0, 10/1) to afford [2-[(1S)-1-[[(2S,4R)-4-acetoxy-1-10 [(2S)-3-methyl-2-[4-(4-oxobutoxy)triazol-1-yl] butanoyl]pyrrolidine-2-carbonyl]amino]ethyl]-4- fluoro-5-(4-methylthiazol-5-yl)phenyl] acetate Int 5l (0.1 g, 145.61 μmol, 83.58% yield) as yellow solid. LCMS: MS (ESI) m/z 687.3 [M+H]⁺ Step M. Preparation of Int 5m To a mixture of Int 5l (67.7 mg, 98.6 μmol, 1.0 eq) and (4S)-2-amino-4-methyl-4-[3-[2-15 [(2S)-2-methyl-1,4-diazepan-1-yl]pyrimidin-4-yl]-1,2,4-oxadiazol-5-yl]-6,7-dihydro-5H- benzothiophene-3-carbonitrile (0.04 g, 88.8 μmol, 0.9 eq) in MeOH (3 mL) was added NaBH3CN (24.8 mg, 395 μmol, 4.0 eq) in one portion at 20°C and stirred for 3 hrs. The mixture was diluted with water (20 mL) and extracted with EtOAc (10 mL x 3). The organic layer was washed with brine, dried over Na₂SO₄, filtered and concentrated. The residue was purified by20 prep-TLC (SiO2, EtOAc:MeOH = 3:1). Compound [2-[(1S)-1-[[(2S,4R)-4-acetoxy-1-[(2S)-2- [4-[4-[(3S)-4-[4-[5-[(4S)-2-amino-3-cyano-4-methyl-6,7-dihydro-5H-benzothiophen-4-yl]- 1,2,4-oxadiazol-3-yl]pyrimidin-2-yl]-3-methyl-1,4-diazepan-1-yl] butoxy]triazol-1-yl]-3- methyl-butanoyl]pyrrolidine-2-carbonyl]amino]ethyl]-4-fluoro-5-(4-methylthiazol-5-yl)phenyl] acetate Int 5m (0.07 g, 62.43 μmol, 63.29% yield) was obtained as a yellow solid. LCMS: MS 25 (ESI) m/z 1121.4 [M+H]⁺ Step N: Preparation of Compound KV-14 To a mixture of Int 5m (0.07 g, 62.4 μmol, 1.0 eq) in MeOH (2 mL) was added K₂CO₃ (129 mg, 936 μmol, 15.0 eq) in one portion at 20°C and stirred for 0.5 hr. The reaction mixture was filtered purified by prep-HPLC(column: Phenomenex luna C18100*40mm*3 um;mobile 30 phase: [H2O(0.1%TFA)-ACN];gradient:20%-55% B over 8.0 min) to KV-14 (0.02 g, 19.28 μmol, 30.89% yield) as yellow solid.¹H NMR (400 MHz, DMSO-d6) δ = 9.85-9.72 (m, 1H), 9.41-9.25 (m, 1H), 9.08 (s, 1H), 8.66 (dd, J = 2.0, 4.8 Hz, 1H), 8.42 (d, J = 8.0 Hz, 1H), 7.73 (s, 1H), 7.27 (dd, J = 4.8, 7.2 Hz, 1H), 7.16-7.08 (m, 2H), 7.04 (d, J = 11.2 Hz, 1H), 6.89-6.77 (m, 1H), 5.19-5.04 (m, 2H), 5.00-4.74 (m, 2H), 4.36 (dd, J = 7.2, 15.6 Hz, 3H), 4.12 (t, J = 5.2 Hz, 35 2H), 3.79-3.67 (m, 2H), 3.66 - 3.60 (m, 1H), 3.59-3.50 (m, 1H), 3.47-3.37 (m, 2H), 3.27-3.19 210 15077.006WO1 (m, 2H), 2.55-2.50 (m, 3H), 2.45-2.38 (m, 2H), 2.34 (s, 3H), 2.15-2.03 (m, 2H), 2.00-1.90 (m, 4H), 1.89-1.82 (m, 4H), 1.79 (s, 3H), 1.77-1.71 (m, 2H), 1.35-1.27 (m, 3H), 1.15 (d, J = 6.0 Hz, 3H), 1.06-0.95 (m, 3H), 0.67 (d, J = 6.4 Hz, 3H). LCMS: MS (ESI) m/z 1037.6 [M+H]⁺ Example KV-17 Synthesis of (2S,4R)-1-[(2S)-2-[4-[4-[(3S)-4-[4-[5-[(4S)-2- 5 amino-3-cyano-4-methyl-6,7-dihydro-5H-benzothiophen-4-yl]-1,2,4-oxadiazol-3-yl]pyrimidin- 2-yl]-3-methyl-1,4-diazepan-1-yl]butoxy]triazol-1-yl]-3-methyl-butanoyl]-4-hydroxy-N-[(1R)- 2-hydroxy-1-[4-[4-[3-(methylamino)prop-1-ynyl]thiazol-5-yl]phenyl]ethyl]pyrrolidine-2- carboxamide, KV-17 10 Step A. Preparation of Int 6a To a solution of (2S,4R)-1-[(2S)-2-azido-3-methyl-butanoyl]-N-[(1R)-1-[4-(4- chlorothiazol-5-yl)phenyl]-2-hydroxy-ethyl]-4-hydroxy-pyrrolidine-2-carboxamide (0.3 g, 608 μmol, 1 eq) tert-butyl-(4-ethynoxybutoxy)-dimethyl-silane (208 mg, 912 μmol, 1.5 eq) in THF 15 (2 mL), t-BuOH (2 mL) and H2O (2 mL) was added copper;sulfate (48.5 mg, 304 μmol, 46.7 μL, 0.5 eq) and sodium;(2R)-2-[(1S)-1,2-dihydroxyethyl]-4-hydroxy-5-oxo-2H-furan-3-olate (180 mg, 912 μmol, 1.5 eq). The mixture was stirred at 20°C for 1 hr. The reaction mixture was 211 15077.006WO1 diluted with H2O (5 mL) and extracted with EtOAc (5 mL x 3). The combined organic layers were washed with brine (10 mL), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The crude product (2S,4R)-1-[(2S)-2-[4-[4-[tert- butyl(dimethyl)silyl]oxybutoxy]triazol-1-yl]-3-methyl-butanoyl]-N-[(1R)-1-[4-(4-chlorothiazol- 5 5-yl)phenyl]-2-hydroxy-ethyl]-4-hydroxy-pyrrolidine-2-carboxamide Int 6a (0.43 g, 596 μmol, 97.9% yield) was used into the next step without further purification. LCMS: MS (ESI) m/z 721.2 [M+H]⁺ Step B. Preparation of Int 6b To a solution of Int 6a (0.9 g, 1.25 mmol, 1 eq), tert-butyl N-methyl-N-prop-2-ynyl- 10 carbamate (633 mg, 3.74 mmol, 3 eq) in MeCN (8 mL) was added K₃PO₄ (794 mg, 3.74 mmol, 3 eq) acetonitrile;dichloropalladium (32.3 mg, 124 μmol, 0.1 eq) and dicyclohexyl-[2-(2,4,6- triisopropylphenyl)phenyl]phosphane (119 mg, 249 μmol, 0.2 eq). The mixture was stirred at 100°C for 16 hrs. The reaction mixture was diluted with H₂O (10 mL) and extracted with EtOAc (10 mL x 2). The combined organic layers were washed with brine (20 mL), dried over 15 Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO₂, Petroleum ether/Ethyl acetate = 100/1 to 0/1). Compound tert-butyl N-[3-[5-[4-[(1R)-1-[[(2S,4R)-1-[(2S)-2-[4-[4-[tert- butyl(dimethyl)silyl]oxybutoxy]triazol-1-yl]-3-methyl-butanoyl]-4-hydroxy-pyrrolidine-2- carbonyl]amino]-2-hydroxy-ethyl]phenyl]thiazol-4-yl]prop-2-ynyl]-N-methyl-carbamate Int 6b 20 (0.42 g, 492 μmol, 39.4% yield) was obtained as a black solid. LCMS: MS (ESI) m/z 854.5 [M+H]⁺ Step C. Preparation of Int 6c To a solution of Int 6b (0.4 g, 468 μmol, 1 eq) in DCM (5 mL) was added Ac₂O (119 mg, 1.17 mmol, 110 μL, 2.5 eq), DMAP (5.72 mg, 46.8 μmol, 0.1 eq) and TEA (189 mg, 1.87 25 mmol, 261 μL, 4 eq). The mixture was stirred at 20°C for 1 hr. The reaction mixture was diluted with H₂O (5 mL) and extracted with DCM (5 mL x 3). The combined organic layers were washed with brine (10 mL), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The crude product [(2R)-2-[[(2S,4R)-4-acetoxy-1-[(2S)-2-[4-[4-[tert- butyl(dimethyl)silyl]oxybutoxy]triazol-1-yl]-3-methyl-butanoyl]pyrrolidine-2-carbonyl]amino]- 30 2-[4-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]thiazol-5-yl]phenyl]ethyl] acetate Int 6c (0.43 g, crude) was used into the next step without further purification. LCMS: MS (ESI) m/z 938.4 [M+H]⁺ Step D. Preparation of Int 6d To a solution of Int 6c (0.3 g, 319 μmol, 1 eq) in MeOH (4 mL) was added acetyl 35 chloride (37.6 mg, 479 μmol, 34.1 μL, 1.5 eq). The mixture was stirred at 20°C for 1 hr. The 212 15077.006WO1 reaction mixture was quenched by addition aq. of NaHCO3 (10 mL), and extracted with EtOAc (10 mL x 2). The combined organic layers were washed with brine (10 mL), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The crude product [(2R)-2- [[(2S,4R)-4-acetoxy-1-[(2S)-2-[4-(4-hydroxybutoxy)triazol-1-yl]-3-methyl- 5 butanoyl]pyrrolidine-2-carbonyl]amino]-2-[4-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1- ynyl]thiazol-5-yl]phenyl]ethyl] acetate Int 6d (0.26 g, crude) was used into the next step without further purification. LCMS: MS (ESI) m/z 824.4 [M+H]⁺ Step E. Preparation of Int 6e To a solution of Int 6d (0.26 g, 315 μmol, 1 eq) in DCM (3 mL) was added Dess-Martin 10 (200 mg, 473 μmol, 146 μL, 1.5 eq). The mixture was stirred at 20°C for 2 hrs. The reaction mixture was quenched by addition aq. of NaHCO3 (10 mL) and extracted with EtOAc (10 mL x 2). The combined organic layers were washed with brine (10 mL), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, Petroleum ether/Ethyl acetate = 100/1 to 0/1). Compound [(2R)-2-15 [[(2S,4R)-4-acetoxy-1-[(2S)-3-methyl-2-[4-(4-oxobutoxy)triazol-1-yl]butanoyl]pyrrolidine-2- carbonyl]amino]-2-[4-[4-[3-[tert-butoxycarbonyl(methyl)amino]prop-1-ynyl]thiazol-5- yl]phenyl]ethyl] acetate Int 6e (0.12 g, 146 μmol, 46.3% yield) was obtained as a white solid. LCMS: MS (ESI) m/z 822.3 [M+H]⁺ 20 To a solution Int 6e (80 mg, 97.3 μmol, 1 eq) and (4S)-2-amino-4-methyl-4-[3-[2-[(2S)- 2-methyl-1,4-diazepan-1-yl]pyrimidin-4-yl]-1,2,4-oxadiazol-5-yl]-6,7-dihydro-5H- benzothiophene-3-carbonitrile (43.8 mg, 97.3 μmol, 1 eq) in DCM (4 mL) was added NaBH(OAc)₃ (61.9 mg, 292 μmol, 3 eq). The mixture was stirred at 20°C for 2 hrs. The 213 15077.006WO1 reaction mixture was quenched by addition aq. of NaHCO3 (10 mL) and extracted with EtOAc (10 mL x 2). The combined organic layers were washed with brine (10 mL), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, Petroleum ether/Ethyl acetate = 100/1 to 0/1). Compound [(2R)- 5 2-[[(2S,4R)-4-acetoxy-1-[(2S)-2-[4-[4-[(3S)-4-[4-[5-[(4S)-2-amino-3-cyano-4-methyl-6,7- dihydro-5H-benzothiophen-4-yl]-1,2,4-oxadiazol-3-yl]pyrimidin-2-yl]-3-methyl-1,4-diazepan-1- yl]butoxy]triazol-1-yl]-3-methyl-butanoyl]pyrrolidine-2-carbonyl]amino]-2-[4-[4-[3-[tert- butoxycarbonyl(methyl)amino]prop-1-ynyl]thiazol-5-yl]phenyl]ethyl] acetate Int 6f (0.1 g, 79.6 μmol, 81.8% yield) was obtained as a white solid. LCMS: MS (ESI) m/z 1256.5 [M+H]⁺ 10 Step G. Preparation of Int 6g To a solution of Int 6f (30 mg, 23.9 μmol, 1 eq) in MeOH (0.5 mL) was added K₂CO₃ (9.90 mg, 71.6 μmol, 3 eq). The mixture was stirred at 20°C for 1 hr. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was diluted 15 with H₂O (2 mL) and extracted with DCM (2 mL x 2). The combined organic layers were dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The crude product tert-butyl N-[3-[5-[4-[(1R)-1-[[(2S,4R)-1-[(2S)-2-[4-[4-[(3S)-4-[4-[5-[(4S)-2-amino-3- cyano-4-methyl-6,7-dihydro-5H-benzothiophen-4-yl]-1,2,4-oxadiazol-3-yl]pyrimidin-2-yl]-3- methyl-1,4-diazepan-1-yl]butoxy]triazol-1-yl]-3-methyl-butanoyl]-4-hydroxy-pyrrolidine-2- 20 carbonyl]amino]-2-hydroxy-ethyl]phenyl]thiazol-4-yl]prop-2-ynyl]-N-methyl-carbamate Int 6g (27 mg, 23.0 μmol, 96.4% yield) was used into the next step without further purification. LCMS: MS (ESI) m/z 1172.4 [M+H]⁺ Step H. Preparation of KV-17 214 15077.006WO1 To a solution of Int 6g (25 mg, 21.3 μmol, 1 eq) in EtOAc (0.2 mL) was added HCl/dioxane (4 M, 0.1 mL, 18.7 eq). The mixture was stirred at 20°C for 1 hr. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by 5 prep-HPLC (column: Phenomenex Luna C1875x30mmx3um; mobile phase: [H2O(0.1% TFA)- ACN];gradient:15%-45% B over 8.0 min) to give KV-17 (4 mg, 3.73 μmol, 17.5% yield) as a white solid.¹ H NMR (400 MHz, DMSO-d6) δ9.12 (s, 1H), 9.06-8.91 (m, 1H), 8.65 (d, J = 4.4 Hz, 1H), 8.51 (d, J = 7.6 Hz, 1H), 7.76 (d, J = 8.0 Hz, 2H), 7.68 (s, 1H), 7.40 (d, J = 8.0 Hz, 2H), 7.26 (s, 1H), 7.09 (s, 2H), 5.17 (d, J = 3.6 Hz, 1H), 5.12 (d, J = 10.4 Hz, 1H), 4.99-4.77 (m, 10 3H), 4.50-4.37 (m, 2H), 4.30 (s, 1H), 4.22 (s, 2H), 4.11 (s, 2H), 3.77-3.70 (m, 1H), 3.66-3.51 (m, 3H), 2.63 (s, 3H), 2.57-2.52 (m, 2H), 2.46-2.35 (m, 3H), 2.14-2.03 (m, 2H), 2.01-1.59 (m, 14H), 1.14 (s, 3H), 1.03 (d, J = 6.4 Hz, 3H), 0.76-0.61 (m, 3H). LCMS: MS (ESI) m/z 1072.5 [M+H]⁺ Example KV-15 Synthesis of (2S,4R)-1-[(2S)-2-[4-[4-[(3S)-4-[4-[5-[(4S)-2-15 amino-3-cyano-4-methyl-6,7-dihydro-5H-benzothiophen-4-yl]-1,2,4-oxadiazol-3-yl]pyrimidin- 215 15077.006WO1 2-yl]-3-methyl-1,4-diazepan-1-yl]butoxy]triazol-1-yl]-3-methyl-butanoyl]-N-[(1S)-1-[3-fluoro- 2-hydroxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]-4-hydroxy-pyrrolidine-2-carboxamide, KV-15 5 Step A. Preparation of Int 7a To a mixture of 1-(4-bromo-3-fluoro-2-hydroxy-phenyl)ethanone (20 g, 85.8 mmol, 1 eq) and benzyl bromide, BnBr (15.4 g, 90.12 mmol, 10.70 mL, 1.05 eq) in acetone (300 mL) was added K2CO3 (14.2 g, 103 mmol, 1.2 eq) in one portion at 20°C under N2. The reaction mixture was heated to 65°C for 12hr. The reaction mixture was quenched with water 300 mL, 10 then the aqueous was extracted with EtOAc (300 mL*3). The combined organic layers were washed with water (300 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, Petroleum ether/Ethyl acetate=1:0 to 3:1). Compound 1-(2-benzyloxy-4-bromo-3-fluoro- phenyl)ethenone Int 7a (25.3 g, 78.3 mmol, 91.34% yield) was obtained as yellow solid.¹ H 15 NMR (DMSO, 400 MHz) δ7.55-7.35 (m, 7H), 5.18 (s, 2H), 2.47 (s, 3H) Step B. Preparation of Int 7b 216 15077.006WO1 To a mixture of Int 7a (20 g, 61.89 mmol, 1 eq) and tetraisopropoxytitanium (87.9 g, 310 mmol, 91.3 mL, 5 eq) in THF (400 mL) was added (S)-2-methylpropane-2-sulfinamide (18.75 g, 155 mmol, 2.5 eq) in one portion at 20°C under N₂. The reaction mixture was heated to 80°C for 16hr. The reaction mixture was quenched with water 200 mL, the aqueous was extracted 5 with EtOAc (300mL*3). The combined organic layers were washed with water (300 mL), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, Petroleum ether/Ethyl acetate=1:0 to 3:1) to give (NE,S)-N-[1-(2-benzyloxy-4-bromo-3-fluoro-phenyl)ethylidene]-2-methyl-propane-2- sulfinamide Int 7b (25 g, 58.6 mmol, 94.75% yield) as a yellow solid. LCMS: MS (ESI) m/z 10 426.0 [M+H]⁺ Step C. Preparation of Int 7c To a mixture of Int 7b (20 g, 46.9 mmol, 1 eq) and CeCl₃ (11.5 g, 46.9 mmol, 1 eq) in THF (400 mL) was added NaBH₄ (7.10 g, 187 mmol, 4.00 eq) in portions at -60°C under N₂. The reaction mixture was warmed to 20 °C for 12hr. The saturated aqueous NH4Cl solution 15 300 mL was added dropwise into the reaction mixture at 0°C under N2, then the reaction mixture was allowed to warm to ambient temperature and extracted with EtOAc (300 mL*3). The combined organic layers were washed with brine (300 mL), dried overNa2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, Petroleum ether/Ethyl acetate=1:0 to 1:1) to give (S)-N-[(1S)-1-(2- 20 benzyloxy-4-bromo-3-fluoro-phenyl)ethyl]-2-methyl-propane-2-sulfinamide Int 7c (18 g, 42.02 mmol, 89.58% yield) as yellow oil. LCMS: MS (ESI) m/z 428.1 [M+H]⁺ Step D. Preparation of Int 7d To a solution of Int 7c (5.00 g, 11.7 mmol, 1 eq) and 4-methylthiazole (3.50 g, 35.0 mmol, 3.20 mL, 3 eq) in DMA (50.0 mL) was added dichloropalladium;triphenylphosphane 25 (820 mg, 1.17 mmol, 0.1 eq) and KOAc (3.40 g, 35.0 mmol, 3 eq) under N2. The mixture was heated to 130°C and stirred for 12 h. The reaction mixture was diluted with H₂O 50 mL and extracted with EtOAc (50 mL x 3). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash30 Column, Eluent of 0~100% Ethyl acetate/Petroleum ether gradient @ 85 mL/min) to give (R)- N-[(1S)-1-[2-benzyloxy-3-fluoro-4-(4-methylthiazol-5-yl)phenyl]ethyl]-2-methyl-propane-2- sulfinamide Int 7d (2.6 g, 5.82 mmol, 49.88% yield) was obtained as a light yellow solid. LCMS: MS (ESI) m/z 447.1 [M+H]⁺ Step E. Preparation of Int 7e 217 15077.006WO1 To a solution of Int 7d (2.60 g, 5.80 mmol, 1 eq) in EtOAc (10.0 mL) was added HCl/EtOAc (4 M, 20.0 mL). The mixture was stirred at 20°C for 1 h. The mixture was filtered and concentrated to give (1S)-1-[2-benzyloxy-3-fluoro-4-(4-methylthiazol-5- yl)phenyl]ethanamine Int 7e (1.3 g, 3.43 mmol, 58.94% yield, HCl) as a light yellow solid.¹ H 5 NMR (MeOD, 400 MHz) δ9.66 (s, 1H), 7.51-7.32 (m, 7H), 5.40-5.31 (m, 1H), 5.30-5.23 (m, 1H), 4.68 (q, J = 6.8 Hz, 1H), 2.49 (s, 3H), 1.45 (d, J = 6.8 Hz, 3H). Step F. Preparation of Int 7f To a solution of Int 7e (1.40 g, 4.20 mmol, 1 eq) in DMF (20.0 mL) was added HATU (1.60 g, 4.20 mmol, 1 eq) and DIEA (2.70 g, 21.0 mmol, 3.70 mL, 5 eq) and (1S)-1-[2- 10 benzyloxy-3-fluoro-4-(4-methylthiazol-5-yl)phenyl]ethanamine (1.30 g, 3.73 mmol, 0.90 eq). The mixture was stirred at 20°C for 1 h. The reaction mixture was diluted with H2O 30 mL and extracted with EtOAc (30 mL x 3). The combined organic layers were washed with brine (20 mL), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash15 Column, Eluent of 0~100% Ethyl acetate/Petroleum ethergradient @ 50 mL/min) to give tert- butyl N-[(1S)-1-[(2S,4R)-2-[[(1S)-1-[2-benzyloxy-3-fluoro-4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]-4-hydroxy-pyrrolidine-1-carbonyl]-2-methyl-propyl]carbamate Int 7f (1.4 g, 2.14 mmol, 50.46% yield) as a light yellow solid. LCMS: MS (ESI) m/z 655.3 [M+H]⁺ Step G. Preparation of Int 7g 20 218 15077.006WO1 To a solution of Int 7f (1.00 g, 1.50 mmol, 1 eq) in DCM (10.0 mL) was added BBr₃ 5 (3.00 g, 12.0 mmol, 1.20 mL, 8 eq) at -78°C under N2. The mixture was warmed to 20°C and stirred at for 4 h. The reaction mixture was quenched by addition H₂O 10 mL at 0°C, and then diluted with H₂O 10 mL and THF (10 mL). The aqueous phase pH adjust to ~8 with NaHCO₃ (aq), then Boc2O (940 mg, 4.30 mmol, 990 μL, 2 eq) was added. The mixture was stirred at 20°C for 1 h. The reaction mixture was extracted with EtOAc (20 mL x 3). The combined 10 organic layers were washed with brine (10 mL), dried over Na₂SO₄, filtered and concentrated 219 15077.006WO1 under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0~100% Ethyl acetate/Petroleum ethergradient @ 45 mL/min) to give tert-butyl N-[(1S)-1-[(2S,4R)-2-[[(1S)-1- [3-fluoro-2-hydroxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]-4-hydroxy-pyrrolidine-1- 5 carbonyl]-2-methyl-propyl]carbamate Int 7g (1 g, 1.77 mmol, 82.27% yield) as a white solid. LCMS: MS (ESI) m/z 565.2 [M+H]⁺ Step H. Preparation of Int 7h To a solution of Int 7g (1.0 g, 1.77 mmol, 1 eq) in EtOAc (10 mL) was added HCl/EtOAc (4 M, 10 mL, 22.6 eq). The mixture was stirred at 25°C for 1 hr. The reaction10 mixture was concentrated under reduced pressure to give a residue. Compound (2S,4R)-1-[(2S)- 2-amino-3-methyl-butanoyl]-N-[(1S)-1-[3-fluoro-2-hydroxy-4-(4-methylthiazol-5- yl)phenyl]ethyl]-4-hydroxy-pyrrolidine-2-carboxamide Int 7h (0.75 g, 1.61 mmol, 91.2% yield) was obtained as a white solid. LCMS: MS (ESI) m/z 465.2 [M+H]⁺ Step I. Preparation of Int 7i 15 To a solution of Int 7h (0.63 g, 1.36 mmol, 1 eq) in MeCN (6 mL) was added 2-azido- 1,3-dimethyl-4,5-dihydroimidazol-1-ium;hexafluorophosphate (464 mg, 1.63 mmol, 1.2 eq) and dropwise DIEA (525 mg, 4.07 mmol, 708 μL, 3 eq) in MeCN (6 mL) at 0°C. The mixture was warmed to 20°C and stirred for 16 hrs. The reaction mixture was quenched by addition H2O (10 mL) and extracted with EtOAc (10 mL). The combined organic layers were washed with brine 20 (10 mL), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, Petroleum ether/Ethyl acetate = 100/1 to 0/1) to give (2S,4R)-1-[(2S)-2-azido-3-methyl-butanoyl]-N-[(1S)-1-[3-fluoro-2- hydroxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]-4-hydroxy-pyrrolidine-2-carboxamide Int 7i (0.5 g, 1.02 mmol, 75.16% yield) as a white solid. LCMS: MS (ESI) m/z 491.2 [M+H]⁺ 25 Step J. Preparation of Int 7j To a solution of Int 7i (0.25 g, 509 μmol, 1 eq), tert-butyl-(4-ethynoxybutoxy)- dimethyl-silane (232 mg, 1.02 mmol, 2 eq) in THF (1 mL), t-BuOH (1 mL) and H₂O (1 mL) was added copper;sulfate (40.6 mg, 254 μmol, 39.1 μL, 0.5 eq) and sodium;(2R)-2-[(1S)-1,2- dihydroxyethyl]-4-hydroxy-5-oxo-2H-furan-3-olate (151 mg, 764 μmol, 1.5 eq). The mixture 30 was stirred at 30°C for 2 hrs. The reaction mixture was diluted with H₂O (5 mL) and extracted with EtOAc (5 mL x 3). The combined organic layers were washed with brine (10 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The crude product (2S,4R)-1-[(2S)-2-[4-[4-[tert-butyl(dimethyl)silyl]oxybutoxy]triazol-1-yl]-3-methyl- butanoyl]-N-[(1S)-1-[3-fluoro-2-hydroxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]-4-hydroxy- 220 15077.006WO1 pyrrolidine-2-carboxamide Int 7j (0.36 g, crude) was used into the next step without further purification. LCMS: MS (ESI) m/z 719.3 [M+H]⁺ Step K. Preparation of Int 7k To a solution of Int 7j (0.42 g, 584 μmol, 1 eq) in DCM (5 mL) was added Ac2O (149 5 mg, 1.46 mmol, 137 μL, 2.5 eq), DMAP (7.14 mg, 58.4 μmol, 0.1eq) and TEA (236 mg, 2.34 mmol, 325 μL, 4 eq). The mixture was stirred at 20°C for 1 hr. The reaction mixture was diluted with H2O (5 mL) and extracted with DCM (5 mL x 3). The combined organic layers were washed with brine (10 mL), dried over [Na2SO4], filtered and concentrated under reduced pressure to give a residue. The crude product [6-[(1S)-1-[[(2S,4R)-4-acetoxy-1-[(2S)-2-[4-[4-10 [tert-butyl(dimethyl)silyl]oxybutoxy]triazol-1-yl]-3-methyl-butanoyl]pyrrolidine-2- carbonyl]amino]ethyl]-2-fluoro-3-(4-methylthiazol-5-yl)phenyl] acetate Int 7k (0.46 g, 572 μmol, 98.1% yield) (yellow oil )was used into the next step without further purification. LCMS: MS (ESI) m/z 803.4 [M+H]⁺ Step L. Preparation of Int 7l 15 To a solution of Int 7k (400 mg, 498.12 μmol, 1 eq) in MeOH (4 mL) was added acetyl chloride (58.6 mg, 747 μmol, 53.1 μL, 1.5 eq). The mixture was stirred at 20°C for 1 hr. The reaction mixture was quenched by addition aq. of NaHCO3 (10 mL), and extracted with EtOAc (10 mL x 2). The combined organic layers were washed with brine (10 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The crude product [6-[(1S)-20 1-[[(2S,4R)-4-acetoxy-1-[(2S)-2-[4-(4-hydroxybutoxy)triazol-1-yl]-3-methyl- butanoyl]pyrrolidine-2-carbonyl]amino]ethyl]-2-fluoro-3-(4-methylthiazol-5-yl)phenyl] acetate Int 7l (0.34 g, 493 μmol, 99.1% yield) was used into the next step without further purification. LCMS: MS (ESI) m/z 689.3 [M+H]⁺ Step M. Preparation of Int 7m 25 To a solution of Int 7l (0.3 g, 435 μmol, 1 eq) in DCM (2 mL)was added Dess-Martin reagent (277 mg, 653 μmol, 202 μL, 1.5 eq). The mixture was stirred at 25°C for 1 hr. The reaction mixture was quenched by addition aq. of NaHCO₃ (10 mL), and extracted with EtOAc (10 mL x 2). The combined organic layers were washed with brine (10 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by30 column chromatography (SiO₂, Petroleum ether/Ethyl acetate = 100/1 to 0/1) to give [6-[(1S)-1- [[(2S,4R)-4-acetoxy-1-[(2S)-3-methyl-2-[4-(4-oxobutoxy)triazol-1-yl]butanoyl]pyrrolidine-2- carbonyl]amino]ethyl]-2-fluoro-3-(4-methylthiazol-5-yl)phenyl] acetate Int 7m (0.13 g, 189 μmol, 43.4% yield) as a white solid. LCMS: MS (ESI) m/z 687.3 [M+H]⁺ Step N. Preparation of Int 7n 221 15077.006WO1 To a solution of Int 7m (120 mg, 174 μmol, 1 eq) and (4S)-2-amino-4-methyl-4-[3-[2- [(2S)-2-methyl-1,4-diazepan-1-yl]pyrimidin-4-yl]-1,2,4-oxadiazol-5-yl]-6,7-dihydro-5H- benzothiophene-3-carbonitrile (78.7 mg, 174 μmol, 1 eq) in MeOH (0.2 mL) was added sodium cyanoboranuide (43.9 mg, 698 μmol, 4 eq). The mixture was stirred at 20°C for 2 hrs. The 5 reaction mixture was diluted with aq. of NaHCO3 (5 mL) and extracted with EtOAc (5 mL x 3). The combined organic layers were washed with brine (10 mL), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: Phenomenex Luna C1875 x 30mm x 3um; mobile phase: [H2O(0.1% TFA)- ACN];gradient:20%-50% B over 8.0 min) to give [6-[(1S)-1-[[(2S,4R)-4-acetoxy-1-[(2S)-2-[4-10 [4-[(3S)-4-[4-[5-[(4S)-2-amino-3-cyano-4-methyl-6,7-dihydro-5H-benzothiophen-4-yl]-1,2,4- oxadiazol-3-yl]pyrimidin-2-yl]-3-methyl-1,4-diazepan-1-yl]butoxy]triazol-1-yl]-3-methyl- butanoyl]pyrrolidine-2-carbonyl]amino]ethyl]-2-fluoro-3-(4-methylthiazol-5-yl)phenyl] acetate Int 7n (0.1 g, 89.1 μmol, 51.04% yield) as a white solid. LCMS: MS (ESI) m/z 1121.4 [M+H]⁺ Step O. Preparation of KV-15 15 To a solution of Int 7n (30 mg, 26.7 μmol, 1 eq) in MeOH (0.3 mL) was added K2CO3 (11.0 mg, 80.26 μmol, 3 eq). The mixture was stirred at 20°C for 1 hr. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified 222 15077.006WO1 by prep-HPLC (column: Phenomenex Luna C1875 x 30mm x 3um;mobile phase: [H2O(0.1% TFA)-ACN];gradient:20%-50% B over 8.0 min) to give KV-15 (7 mg, 6.75 μmol, 25.2% yield) as a white solid.¹H NMR (400 MHz, DMSO-d₆) δ9.99-9.85 (m, 1H), 9.44-9.25 (m, 1H), 9.09 (s, 1H), 8.66 (d, J = 2.4 Hz, 1H), 8.47 (d, J = 7.2 Hz, 1H), 7.76-7.63 (m, 1H), 7.26 (d, J = 5.9 Hz, 5 1H), 7.17-6.97 (m, 2H), 6.93-6.76 (m, 1H), 5.31-5.05 (m, 2H), 5.00-4.74 (m, 1H), 4.52-4.26 (m, 3H), 4.20-3.98 (m, 2H), 3.87-3.68 (m, 3H), 3.24-3.20 (m, 3H), 2.69-2.64 (m, 1H), 2.39-2.27 (m, 4H), 2.15-1.60 (m, 16H), 1.33 (d, J = 6.9 Hz, 3H), 1.21-0.98 (m, 6H), 0.94 (d, J = 6.6 Hz, 1H), 0.74-0.57 (m, 3H). MS (ESI) m/z 1037.5[M+H]⁺ The following compounds were prepared in a manner similar to that described for KV-5 10 using the appropriate compound as starting material. 223 15077.006WO1 Example KV-18 Synthesis of (2S,4R)-1-[(2R)-2-[4-[4-[(3S)-4-[4-[5-[(4S)- 2-amino-3-cyano-4-methyl-6,7-dihydro-5H-benzothiophen-4-yl]-1,2,4-oxadiazol-3- yl]pyrimidin-2-yl]-3-methyl-1,4-diazepan-1-yl]butoxy]triazol-1-yl]-3-methyl-3-[3-5 (methylamino) propylsulfanyl]butanoyl]-4-hydroxy-N-[(1R)-2-hydroxy-1-[4-(4-methylthiazol- 5-yl)phenyl] ethyl]pyrrolidine-2-carboxamide, KV-18 224 15077.006WO1 Step A: KV-18a 5 To a mixture of 3-chloro-N-methyl-propan-1-amine (3 g, 20.8 mmol, 1.0 eq, HCl) in THF (50 mL) and H₂O (10 mL) were added allyl carbonochloridate (2.51 g, 20.8 mmol, 2.21 mL, 1.0 eq) and NaHCO₃ (3.50 g, 41.7 mmol, 1.62 mL, 2.0 eq) at 20°C and stirred for 1 hr. The residue was poured into ice-water (w/w = 1/1) (50 mL) and stirred for 2 min. The aqueous phase was extracted with ethyl acetate (30 mL x 3). The combined organic phase was washed 10 with brine (30 mL x 2), dried with anhydrous Na₂SO₄, filtered and concentrated in vacuum to 225 15077.006WO1 give allyl N-(3-chloropropyl)-N-methyl-carbamate, KV-18a (3.8 g, crude) as colorless oil.¹H NMR (400 MHz, DMSO-d₆) δ = 6.01- 5.84 (m, 1H), 5.34-5.13 (m, 2H), 4.57-4.43 (m, 2H), 3.63 (t, J = 6.4 Hz, 2H), 3.32 (s, 2H), 2.85 (d, J = 10.8 Hz, 3H), 1.97–1.88 (m, 2H). Step B: KV-18b 5 To a mixture of KV-18a (1.1 g, 5.74 mmol, 1.0 eq) and (2R)-2-(tert- butoxycarbonylamino)-3-methyl-3-sulfanyl-butanoic acid (1.43 g, 5.74 mmol, 1.0 eq) in DMF (20 mL) was added Cs2CO3 (3.74 g, 11.5 mmol, 2.0 eq) at 20°C and stirred for 16 hrs. The mixture was diluted with water (50 mL) and extracted with EtOAc (30 mL x 3). The organic layer was washed with brine, dried over Na₂SO₄, filtered and concentrated. The residue was 10 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 afford (2R)-3-[3- [allyloxycarbonyl(methyl)amino]propylsulfanyl]-2-(tert-butoxycarbonylamino)-3-methyl- butanoic acid, KV-18b (1.9 g, 4.70 mmol, 81.84% yield) as colorless oil.¹H NMR (400 MHz, DMSO-d6) δ = 12.71 (s, 1H), 6.90 (d, J = 9.2 Hz, 1H), 5.99-5.85 (m, 1H), 5.34-5.13 (m, 2H), 15 4.54-4.45 (m, 2H), 4.10 (d, J = 9.2 Hz, 1H), 3.26 (d, J = 1.6 Hz, 2H), 2.83 (d, J = 11.2 Hz, 3H), 1.66 (s, 2H), 1.38 (s, 9H), 1.32 (s, 3H), 1.25 (s, 3H). LCMS: MS (ESI) m/z 405.3 [M+H]⁺ Step C: KV-18c To a mixture of KV-18b (1 g, 2.47 mmol, 1.0 eq) in DMF (10 mL) were added HATU (940 mg, 2.47 mmol, 1.0 eq), DIEA (1.60 g, 12.4 mmol, 2.15 mL, 5.0 eq) and (2S,4R)-4-20 hydroxy-N-[(1R)-2-hydroxy-1-[4-(4-methylthiazol-5-yl)phenyl]ethyl] pyrrolidine-2- carboxamide (1.23 g, 3.21 mmol, 1.3 eq, HCl) at 20°C and stirred for 1 hr. The mixture was diluted with water (30 mL) and extracted with EtOAc(30 mL x 3). The organic layer was washed with brine, drid over Na₂SO₄, filtere and concentrated. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel,25 Petroleum ether/Ethyl acetate=1/0, 0/1) to afford allyl N-[3-[(2R)-2-(tert-butoxycarbonylamino)- 3-[(2S,4R)-4-hydroxy-2-[[(1R)-2-hydroxy-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidin-1-yl]-1,1-dimethyl-3-oxo-propyl]sulfanylpropyl]-N- methyl-carbamate, KV-18c (0.85 g, 1.16 mmol, 46.85% yield) as yellow oil.¹H NMR (400 MHz, DMSO-d₆) δ = 8.98 (s, 1H), 8.28 (d, J = 7.6 Hz, 1H), 7.49-7.28 (m, 4H), 6.75 (d, J = 9.6 30 Hz, 1H), 6.00 - 5.82 (m, 1H), 5.26 (dd, J = 1.6, 17.2 Hz, 1H), 5.20-5.12 (m, 2H), 4.90-4.75 (m, 2H), 4.54-4.41 (m, 4H), 4.30 (s, 1H), 3.71-3.54 (m, 4H), 3.26 (s, 2H), 2.83 (d, J = 8.4 Hz, 3H), 2.45 (s, 3H), 2.13-2.03 (m, 1H), 1.89-1.78 (m, 1H), 1.66 (s, 2H), 1.42-1.38 (m, 9H), 1.37 (s, 3H), 1.24 (s, 3H). LCMS: MS (ESI) m/z 734.5 [M+H]⁺. Step D: KV-18d 226 15077.006WO1 To a mixture of KV-8c (0.85 g, 1.16 mmol, 1.0 eq) in EtOAc (5 mL) was added HCl/EtOAc (20 mL) at 20°C and stirred at 20°C for 0.5 hr. The mixture was concentrated. Compound allyl N-[3-[(2R)-2-amino-3-[(2S,4R)-4-hydroxy-2-[[(1R)-2-hydroxy-1-[4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl] pyrrolidin-1-yl]-1,1-dimethyl-3-oxo- 5 propyl]sulfanylpropyl]-N-methyl-carbamate, KV-18d (0.8 g, crude, HCl) was obtained as a yellow solid. LCMS: MS (ESI) m/z 634.3 [M+H]⁺. Step E: KV-18e To a solution of KV-18d (0.7 g, 1.04 mmol, 1 eq, HCl) and 2-azido-1,3-dimethyl-4,5- dihydroimidazol-1-ium;hexafluorophosphate (596 mg, 2.09 mmol, 2 eq) in MeCN (6 mL) was 10 added a solution of DIEA (405 mg, 3.13 mmol, 546 μL, 3.0 eq) in MeCN (5 mL) dropwise at 0°C over a period of 5 min under N2. Then the mixture solution was stirred at 0°C for 1 hr. The residue was poured into ice-water (w/w = 1/1) (50 mL) and stirred for 2 min. The aqueous phase was extracted with ethyl acetate (30 mL x 3). The combined organic phase was washed with brine (30 mL x 2), dried with anhydrous Na2SO4, filtered and concentrated in vacuum. The 15 residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, Ethyl acetate/MeOH=1/0, 10/1) to afford allyl N-[3-[(2R)-2-azido-3- [(2S,4R)-4-hydroxy-2-[[(1R)-2-hydroxy-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidin-1-yl]-1,1-dimethyl-3-oxo-propyl]sulfanylpropyl]-N- methyl-carbamate, KV-18e (0.35 g, 530.45 μmol, 50.79% yield) as yellow solid. LCMS: MS 20 (ESI) m/z 660.4 [M+H]⁺. Step F: KV-18f To a mixture of KV-18e (0.32 g, 485 μmol, 1.0 eq) and tert-butyl-(4-ethynoxybutoxy)- dimethyl-silane (144 mg, 630 μmol, 1.3 eq) in THF (1.5 mL), H₂O (1.5 mL) and t-BuOH (1.5 mL) were added sodium;(2R)-2-[(1S)-1,2-dihydroxyethyl]-4-hydroxy-5-oxo-2H-furan-3-olate 25 (144 mg, 727 μmol, 1.5 eq) and CuSO4 (38.7 mg, 242 μmol, 37.2 μL, 0.5 eq) in one portion at 20°C under N₂. The mixture was stirred at 30°C for 2 hrs. The residue was poured into ice- water (w/w = 1/1) (30 mL) and stirred for 2 min. The aqueous phase was extracted with ethyl acetate (20 mL x 3). The combined organic phase was washed with brine (20 mL x 2), dried with anhydrous Na₂SO₄, filtered and concentrated in vacuum to give allyl N-[3-[(2R)-2-[4-[4-30 [tert-butyl(dimethyl)silyl]oxybutoxy]triazol-1-yl]-3-[(2S,4R)-4-hydroxy-2-[[(1R)-2-hydroxy-1- [4-(4-methylthiazol-5-yl)phenyl]ethyl] carbamoyl]pyrrolidin-1-yl]-1,1-dimethyl-3-oxo- propyl]sulfanylpropyl]-N-methyl-carbamate, KV-18f (0.4 g, crude) as yellow oil. LCMS: MS (ESI) m/z 888.5 [M+H]⁺. Step G: KV-18g 227 15077.006WO1 228 15077.006WO1 To a mixture of KV-18f (0.4 g, 450 μmol, 1.0 eq) in DCM (6 mL) was added DMAP (5.50 mg, 45.0 μmol, 0.1 eq) Ac₂O (115 mg, 1.13 mmol, 106 μL, 2.5 eq) and Et₃N (91.1 mg, 901 μmol, 125 μL, 2.0 eq) in one portion at 20°C and stirred for 1 hr. The mixture was diluted with water and extracted with DCM (20 mL x 3). The organic layer was washed with brine, dried 5 over Na2SO4, filtered and concentrated. Compound [(2R)-2-[[(2S,4R)-4-acetoxy-1-[(2R)-3-[3- [allyloxycarbonyl(methyl)amino]propylsulfanyl]-2-[4-[4-[tert-butyl(dimethyl)silyl] oxybutoxy]triazol-1-yl]-3-methyl-butanoyl]pyrrolidine-2-carbonyl]amino]-2-[4-(4- methylthiazol-5-yl)phenyl]ethyl] acetate, KV-18g (0.35 g, crude) was obtained as yellow oil. LCMS: MS (ESI) m/z 972.3 [M+H]⁺. 10 Step H: KV-18h To a mixture of KV-18g (0.35 g, 360 μmol, 1.0 eq) in MeOH (6 mL) was added acetyl chloride (42.4 mg, 540 μmol, 38.4 μL, 1.5 eq) at 20°C and stirred for 0.5 hr. The mixture was added aq.NaHCO₃ (10 mL). extracted with EtOAc (30 mL x 3). The organic layer was washed with brine, dried over Na2SO4, filtered and concentrated to give [(2R)-2-[[(2S,4R)-4-acetoxy-1-15 [(2R)-3-[3-[allyloxycarbonyl(methyl)amino]propylsulfanyl]-2-[4-(4-hydroxybutoxy)triazol-1- yl]-3-methyl-butanoyl]pyrrolidine-2-carbonyl]amino]-2-[4-(4-methylthiazol-5-yl)phenyl]ethyl] acetate, KV-18h (0.34 g, crude) as yellow oil. LCMS: MS (ESI) m/z 858.2 [M+H]⁺. Step I: KV-8i To a mixture of KV-18h (0.32 g, 373 μmol, 1.0 eq) in DCM (5 mL) was added Dess- 20 Martin (253mg, 597 μmol, 185 μL, 1.6 eq) in one portion at 20°C and stirred at 20°C for 0.5 hr. The mixture was filtered and concentrated. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, Ethyl acetate/MeOH=1/0, 10/1) to afford [(2R)-2-[[(2S,4R)-4-acetoxy-1-[(2R)-3-[3- [allyloxycarbonyl(methyl)amino]propylsulfanyl]-3-methyl-2-[4-(4-oxobutoxy)triazol-1- 25 yl]butanoyl]pyrrolidine-2-carbonyl]amino]-2-[4-(4-methylthiazol-5-yl)phenyl]ethyl] acetate, KV-18i (0.22 g, 257.00 μmol, 68.91% yield) as yellow solid. LCMS: MS (ESI) m/z 856.3 [M+H]⁺. Step J: KV-18j To a mixture of KV-18i (0.2 g, 233.64 μmol, 1 eq) and (4S)-2-amino-4-methyl-4-[3-[2-30 [(2S)-2-methyl-1,4-diazepan-1-yl]pyrimidin-4-yl]-1,2,4-oxadiazol-5-yl]-6,7-dihydro-5H- benzothiophene-3-carbonitrile (84.2 mg, 187 μmol, 0.8 eq) in DCM (4 mL) was added NaBH(OAc)3 (198 mg, 935 μmol, 4.0 eq) in one portion at 20°C and stirred at 20°C for 0.5 hr. The residue was poured into ice-water (w/w = 1/1) (30 mL) and stirred for 2 min. The aqueous phase was extracted with ethyl acetate (20 mL x 3). The combined organic phase was washed 229 15077.006WO1 with brine (20 mL x 2), dried with anhydrous Na2SO4, filtered and concentrated in vacuum. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, Ethyl acetate/MeOH=1/0, 3/1) to afford [(2R)-2-[[(2S,4R)-4-acetoxy-1- [(2R)-3-[3-[allyloxycarbonyl(methyl)amino]propylsulfanyl]-2-[4-[4-[(3S)-4-[4-[5-[(4S)-2- 5 amino-3-cyano-4-methyl-6,7-dihydro-5H-benzothiophen-4-yl]-1,2,4-oxadiazol-3-yl]pyrimidin- 2-yl]-3-methyl-1,4-diazepan-1-yl]butoxy]triazol-1-yl]-3-methyl-butanoyl]pyrrolidine-2- carbonyl] amino]-2-[4-(4-methylthiazol-5-yl)phenyl]ethyl] acetate, KV-18j (0.22 g, 170.47 μmol, 72.96% yield) as yellow solid. LCMS: MS (ESI) m/z 1290.4 [M+H]⁺. Step K: KV-18k 10 To a mixture of KV-8j (0.22 g, 170 μmol, 1.0 eq) in MeOH (0.5 mL) was added K₂CO₃ (353 mg, 2.56 mmol, 15 eq) in one portion at 20°C and stirred for 0.5 hr. The mixture was filtered and purified by prep-HPLC(column: Phenomenex Luna C1875*30mm*3um;mobile phase: [H₂O(0.1% TFA)-ACN];gradient:20%-60% B over 8.0 min ) to give allyl N-[3-[(2R)-2- [4-[4-[(3S)-4-[4-[5-[(4S)-2-amino-3-cyano-4-methyl-6,7-dihydro-5H-benzothiophen-4-yl]-15 1,2,4-oxadiazol-3-yl]pyrimidin-2-yl]-3-methyl-1,4-diazepan-1-yl]butoxy]triazol-1-yl]-3- [(2S,4R)-4-hydroxy-2-[[(1R)-2-hydroxy-1-[4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidin-1-yl]-1,1-dimethyl-3-oxo-propyl]sulfanylpropyl]-N- methyl-carbamate, KV-8k (0.1 g, 82.88 μmol, 48.62% yield) as yellow solid. LCMS: MS (ESI) m/z 1206.4 [M+H]⁺. 20 Step L: KV-18 To a mixture of KV-18k (0.07 g, 58.0 μmol, 1.0 eq) in THF (3 mL) were added 5,5- dimethylcyclohexane-1,3-dione (61.8 mg, 441 μmol, 7.6 eq) and Pd(PPh₃)₄ (6.70 mg, 5.80 μmol, 0.1 eq) in one portion at 20°C under N₂ and stirred for 0.5 hr. The mixture was filtered and purified by prep-HPLC(olumn: Phenomenex Luna C1875*30mm* 3um;mobile phase: 25 [H2O(0.1% TFA)-ACN];gradient:10%-40% B over 8.0 min ) to give KV-18 (37 mg, 32.96 μmol, 56.82% yield) as yellow solid.¹H NMR (400 MHz, DMSO-d₆) δ = 8.99 (s, 1H), 8.66 (d, J = 4.8 Hz, 1H), 8.50 (d, J = 7.6 Hz, 1H), 7.84 (s, 1H), 7.48-7.41 (m, 2H), 7.38 (d, J = 8.4 Hz, 2H), 7.32-7.25 (m, 1H), 5.60-5.44 (m, 1H), 4.95-4.77 (m, 2H), 4.52-4.36 (m, 2H), 4.35-4.24 (m, 1H), 4.18-4.07 (m, 2H), 3.78-3.54 (m, 6H), 3.47-3.40 (m, 2H), 3.23 (s, 2H), 2.92 (t, J = 7.6 Hz, 30 2H), 2.56 (s, 3H), 2.46 (s, 3H), 2.16-2.03 (m, 2H), 2.02-1.82 (m, 8H), 1.79 (s, 3H), 1.77-1.70 (m, 4H), 1.51-1.44 (m, 3H), 1.38 (s, 3H), 1.14 (s, 3H). LCMS: MS (ESI) m/z 1122.5 [M+H]⁺. Example KV-22 Synthesis of [(3R,5S)-1-[(2S)-2-[4-[4-[(3S)-4-[4-[5-[(4S)-2- amino-3-cyano-4-methyl-6,7-dihydro-5H-benzothiophen-4-yl]-1,2,4-oxadiazol-3-yl]pyrimidin- 2-yl]-3-methyl-1,4-diazepan-1-yl]butoxy]triazol-1-yl]-3-methyl-butanoyl]-5-[[(1S)-1-[5-fluoro- 230 15077.006WO1 2-hydroxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidin-3-yl] dihydrogen phosphate, KV-22 5 Step A: KV-22a To a solution of benzyl (2S,4R)-1-[(2S)-2-[4-[4-[tert- butyl(dimethyl)silyl]oxybutoxy]triazol-1-yl]-3-methyl-butanoyl]-4-hydroxy-pyrrolidine-2- carboxylate (1.2 g, 2.09 mmol, 1 eq) in DMF (10 mL) were added 2H-tetrazole (1.46 g, 20.8 mmol, 1.85 mL, 10 eq) and N-ditert-butoxyphosphanyl-N-ethyl-ethanamine (5.21 g, 20.8 mmol, 10 10 eq). The reaction mixture was stirred at 25°C for 0.5hr. Then H2O2 (2.6 g, 22.9 mmol, 2.20 mL, 30% purity, 10.9 eq) was added dropwise at 0°C. The reaction mixture was stirred and warmed at 20 °C for 0.5hr. The reactions were quenched with aqueous H₂O (20 mL) at 10°C, and then stirred for 2min. The precipitate was filtered, and the filtrate was extracted with EtOAc (3×10 mL). The combined organic layers were dried (Na2SO4), filtered, concentrated. The 15 residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 100/1 to 1/1) to give benzyl (2S,4R)-1-[(2S)-2-[4-[4-[tert-butyl(dimethyl)silyl]oxybutoxy]triazol-1-yl]- 231 15077.006WO1 3-methyl-butanoyl]-4-ditert-butoxyphosphoryloxy-pyrrolidine-2-carboxylate, KV-22a (1.2 g, 1.56 mmol, 74.84% yield) was obtained as a colorless oil. LCMS: MS (ESI) m/z 767.3 [M+H]⁺. Step B: KV-22b To a solution of KV-22a (0.5 g, 651 μmol, 1 eq) in THF (5 mL) was added TBAF (1 M, 5 2.61 mL, 4 eq). The reaction mixture was stirred at 20°C for 1hr. The reaction mixture was diluted with H₂O (20 mL) and extracted with EtOAc (20 mL x 3). The combined organic layers was washed with brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to dryness. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 100/1 to 0/1) to give benzyl (2S,4R)-4-ditert-10 butoxyphosphoryloxy-1-[(2S)-2-[4-(4-hydroxybutoxy)triazol-1-yl]-3-methyl- butanoyl]pyrrolidine-2-carboxylate, KV-22b (0.2 g, 306.41 μmol, 47.00% yield) was obtained as a yellow oil. LCMS: MS (ESI) m/z 653.2 [M+H]⁺. Step C: KV-22c To a solution of KV-22b (0.2 g, 306 μmol, 1 eq) in DCM (2 mL) was added Dess-Martin 15 (324 mg, 766 μmol, 237 μL, 2.5 eq) at 0°C. The reaction mixture was stirred at 20°C for 2hr. The mixture was adjusted to pH = 7 with saturated aqueous NaHCO₃ and extracted with EtOAc (5 mL x 3). The combined organic phase was washed with brine (5 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure to dryness. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 100/1 to 0/1) to give20 benzyl (2S,4R)-4-ditert-butoxyphosphoryloxy-1-[(2S)-3-methyl-2-[4-(4-oxobutoxy)triazol-1- yl]butanoyl]pyrrolidine-2-carboxylate, KV-22c (0.1 g, 153 μmol, 50.1% yield) as a yellow solid. LCMS: MS (ESI) m/z 651.3 [M+H]⁺. Step D: KV-22d To a solution of KV-22c (0.09 g, 138.31 μmol, 1 eq) in DCM (1 mL) was added 25 NaBH(OAc)3 (117 mg, 553 μmol, 4 eq) and (4S)-2-amino-4-methyl-4-[3-[2-[(2S)-2-methyl-1,4- diazepan-1-yl]pyrimidin-4-yl]-1,2,4-oxadiazol-5-yl]-6,7-dihydro-5H-benzothiophene-3- carbonitrile (49.8 mg, 110 μmol, 0.8 eq) in one portion. The reaction mixture was stirred at 25°C for 1hr. The reaction mixture was diluted with H2O (10 mL) and extracted with DCM (5 mL x 3). The combined organic layers were washed with brine (5 mL), dried over anhydrous 30 Na₂SO₄, filtered and concentrated under reduced pressure to dryness. The residue was purified by prep-TLC (SiO2, EtOAc:MeOH = 5:1) to give benzyl (2S,4R)-1-[(2S)-2-[4-[4-[(3S)-4-[4-[5- [(4S)-2-amino-3-cyano-4-methyl-6,7-dihydro-5H-benzothiophen-4-yl]-1,2,4-oxadiazol-3- yl]pyrimidin-2-yl]-3-methyl-1,4-diazepan-1-yl]butoxy]triazol-1-yl]-3-methyl-butanoyl]-4-ditert- butoxyphosphoryloxy-pyrrolidine-2-carboxylate, KV-22d (0.1 g, 92.14 μmol, 66.6% yield) was 35 obtained as a yellow solid. LCMS: MS (ESI) m/z 1085.3 [M+H]⁺ 232 15077.006WO1 Step E: KV-22e To a solution of KV-22d (0.07 g, 64.5 μmol, 1 eq) in THF (0.2 mL) and MeOH (0.2 mL) 5 and H2O (0.2 mL) was added LiOH.H2O (8.12 mg, 193 μmol, 3 eq). The reaction mixture was stirred at 20°C for 1hr. The reaction mixture was concentrated under reduced pressure to dryness. The residue was purified by prep-HPLC (column: Phenomenex Luna C1875 x 30mm x 3um; mobile phase: [H₂O (0.1% TFA)-ACN]; gradient: 25%-60% B over 8.0 min) to give (2S,4R)-1-[(2S)-2-[4-[4-[(3S)-4-[4-[5-[(4S)-2-amino-3-cyano-4-methyl-6,7-dihydro-5H-10 benzothiophen-4-yl]-1,2,4-oxadiazol-3-yl]pyrimidin-2-yl]-3-methyl-1,4-diazepan-1- yl]butoxy]triazol-1-yl]-3-methyl-butanoyl]-4-ditert-butoxyphosphoryloxy-pyrrolidine-2- carboxylic acid, KV-22e (30 mg, 30.1 μmol, 46.7% yield) as a white solid. LCMS: MS (ESI) m/z 995.4 [M+H]⁺. Step F: KV-22f 15 To a solution of KV-22e (0.03 g, 30.1 μmol, 1 eq) in DCM (0.5 mL) was added EDCI (23.1 mg, 120 μmol, 4 eq) and 2,3,5,6-tetrafluorophenol (20.0 mg, 120 μmol, 4 eq). The 233 15077.006WO1 reaction mixture was stirred at 25°C for 1hr. The reaction mixture was diluted with H2O (10 mL) and extracted with DCM (5 mL x 3). The combined organic layers was washed with brine (5 mL), dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to dryness to give (2,3,5,6-tetrafluorophenyl) (2S,4R)-1-[(2S)-2-[4-[4-[(3S)-4-[4-[5-[(4S)-2-amino- 5 3-cyano-4-methyl-6,7-dihydro-5H-benzothiophen-4-yl]-1,2,4-oxadiazol-3-yl]pyrimidin-2-yl]-3- methyl-1,4-diazepan-1-yl]butoxy]triazol-1-yl]-3-methyl-butanoyl]-4-ditert- butoxyphosphoryloxy-pyrrolidine-2-carboxylate (30 mg, crude) as a yellow solid. To a solution of (2,3,5,6-tetrafluorophenyl) (2S,4R)-1-[(2S)-2-[4-[4-[(3S)-4-[4-[5-[(4S)- 2-amino-3-cyano-4-methyl-6,7-dihydro-5H-benzothiophen-4-yl]-1,2,4-oxadiazol-3-10 yl]pyrimidin-2-yl]-3-methyl-1,4-diazepan-1-yl]butoxy]triazol-1-yl]-3-methyl-butanoyl]-4-ditert- butoxyphosphoryloxy-pyrrolidine-2-carboxylate (30 mg, 26.2 μmol, 1 eq) in DMF (0.5 mL) was added DIEA (10.1 mg, 78.7 μmol, 13.7 μL, 3 eq) and 2-[(1S)-1-aminoethyl]-4-fluoro-5-(4- methylthiazol-5-yl)phenol (9.09 mg, 31.4 μmol, 1.2 eq, HCl). The reaction mixture was stirred at 25°C for 1hr. The reaction mixture was diluted with H2O (5 mL) and extracted with DCM (5 15 mL x 3). The combined organic layers were washed with brine (5 mL), dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to dryness to give [(3R,5S)-1-[(2S)-2- [4-[4-[(3S)-4-[4-[5-[(4S)-2-amino-3-cyano-4-methyl-6,7-dihydro-5H-benzothiophen-4-yl]- 1,2,4-oxadiazol-3-yl]pyrimidin-2-yl]-3-methyl-1,4-diazepan-1-yl]butoxy]triazol-1-yl]-3-methyl- butanoyl]-5-[[(1S)-1-[5-fluoro-2-hydroxy-4-(4-methylthiazol-5- 20 yl)phenyl]ethyl]carbamoyl]pyrrolidin-3-yl] ditert-butyl phosphate, KV-22f (30 mg, crude) was obtained as a yellow solid. LCMS: MS (ESI) m/z 1229.3 [M+H]⁺. Step G: KV-22 To a solution of KV-22f (0.03 g, 24.4 μmol, 1 eq) in DCM (0.3 mL) was added TFA (1 mL). The reaction mixture was stirred at 25°C for 1hr. The reaction mixture was concentrated 25 under reduced pressure to dryness. The residue was purified by prep-HPLC (column: Phenomenex Luna C1875 x 30mm x 3um; mobile phase: [H₂O (0.1% TFA)-ACN]; gradient: 25%-55% B over 8.0 min) to give KV-22 (3.4 mg, 2.92 μmol, 11.9% yield, 95.9% purity) as a white solid.¹H NMR (400 MHz, DMSO-d6) δ9.06 (s, 1H), 8.65 (s, 1H), 8.52 (d, J = 8.0 Hz, 1H), 7.72 (s, 1H), 7.26 (s, 1H), 7.04 (d, J = 11.2 Hz, 1H), 6.83 (d, J = 6.4 Hz, 1H), 5.14 (d, J =30 10.4 Hz, 1H), 5.11-5.05 (m, 1H), 4.95-4.77 (m, 2H), 4.39 (t, J = 8.4 Hz, 2H), 4.11 (s, 2H), 3.94- 3.81 (m, 2H), 3.75-3.62 (m, 1H), 3.44-3.34 (m, 2H), 3.22 (s, 2H), 2.71-2.64 (m, 1H), 2.56-2.52 (m, 2H), 2.45-2.35 (m, 2H), 2.33 (s, 3H), 2.12-2.01 (m, 1H), 1.97-1.80 (m, 7H), 1.78 (s, 3H), 1.76-1.68 (m, 3H), 1.43-1.25 (m, 3H), 1.07-0.99 (m, 3H), 0.68 (d, J = 6.4 Hz, 3H). LCMS: MS (ESI) m/z 1117.4[M+H]⁺. 234 15077.006WO1 Example KV-23 Synthesis of (2S,4R)-1-[(2S)-2-[4-[4-[(3S)-4-[4-[5-[(4S)-2-amino- 3-cyano-4-methyl-6,7-dihydro-5H-benzothiophen-4-yl]-1,2,4-oxadiazol-3-yl]pyrimidin-2-yl]-3- methyl-1,4-diazepan-1-yl]butoxy]triazol-1-yl]-3-methyl-butanoyl]-N-[(1S)-1-[4-(2,5-difluoro-4- hydroxy-phenyl)phenyl]ethyl]-4-hydroxy-pyrrolidine-2-carboxamide, KV-23 5 Step A: KV-23a To a mixture of tert-butyl N-[(1S)-1-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2- yl)phenyl] ethyl]carbamate (0.3 g, 864 μmol, 1.0 eq) and 4-bromo-2,5-difluoro-phenol (271 mg, 1.30 mmol, 1.5 eq) in dioxane (5 mL) and H₂O (1 mL) were added K₂CO₃ (299 mg, 2.16 mmol, 10 2.5 eq) and Pd(dppf)Cl2 (63.2 mg, 86.4 μmol, 0.1 eq) in one portion at 20°C under N2. Then the mixture was heated to 100°C and stirred for 2 hrs. The mixture was cooled to 20°C and poured into ice-water (w/w = 1/1) (30 mL) and stirred for 2 min. The aqueous phase was extracted with ethyl acetate (15 mL x 3). The combined organic phase was washed with brine (15 mL x 2), dried with anhydrous Na2SO4, filtered and concentrated in vacuum. The residue was purified by 15 silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, Petroleum ether/Ethyl acetate=1/0, 5/1) to afford tert-butyl N-[(1S)-1-[4-(2,5-difluoro-4- hydroxy-phenyl)phenyl]ethyl]carbamate, KV-23a (0.25 g, 715.57 μmol, 82.83% yield) as yellow solid. LCMS: MS (ESI) m/z 294.2 [M-55]⁺. Step B: KV-23b 20 To a mixture of KV-13a (0.25 g, 716 μmol, 1.0 eq) in EtOAc (2 mL) was added HCl/EtOAc (4 M, 4 mL, 22.36 eq) in one portion at 20°C and stirred at 20°C for 2 hrs. The mixture was concentrated. The crude product was triturated with MTBE at 20^oC for 30 min to give 4-[4-[(1S)-1-aminoethyl]phenyl]-2,5-difluoro-phenol, KV-23b (0.2 g, crude, HCl) as a white solid.¹H NMR (400 MHz, DMSO-d6) δ = 7.56 (s, 4H), 7.37 (dd, J = 7.6, 11.6 Hz, 1H), 25 6.91 (dd, J = 7.6, 11.6 Hz, 1H), 4.43 (q, J = 6.4 Hz, 1H), 1.53 (d, J = 6.8 Hz, 3H). Step C: KV-23 235 15077.006WO1 To a mixture of KV-23b (9 mg, 31.5 μmol, 1.0 eq, HCl) in DMF (0.5 mL) was added DIEA (12.2 mg, 94.5 μmol, 16.5 μL, 3.0 eq) and (2,3,5,6-tetrafluorophenyl) (2S,4R)-1-[(2S)-2- [4-[4-[(3S)-4-[4-[5-[(4S)-2-amino-3-cyano-4-methyl-6,7-dihydro-5H-benzothiophen-4-yl]- 5 1,2,4-oxadiazol-3-yl]pyrimidin-2-yl]-3-methyl-1,4-diazepan-1-yl]butoxy]triazol-1-yl]-3-methyl- butanoyl]-4-hydroxy-pyrrolidine-2-carboxylate (59.9 mg, 63.0 μmol, 2.0 eq) at 20°C and stirred at 20°C for 0.5 hr. The mixture was purified by prep-HPLC(column: WePure Biotech XP tC18 150*40*70um;mobile phase: [H2O(10mM NH4HCO3)-ACN];gradient:55%-85% B over 8.0 min ) to give KV-23 (7.1 mg, 6.87 μmol, 21.79% yield) as white solid.¹H NMR (400 MHz, 10 DMSO-d6) δ = 8.61 - 8.51 (m, 1H), 8.47 (d, J = 7.6 Hz, 1H), 7.71 - 7.64 (m, 1H), 7.48 - 7.40 (m, 2H), 7.37 - 7.28 (m, 3H), 7.14 - 7.03 (m, 3H), 6.90 - 6.80 (m, 1H), 5.15 (d, J = 3.2 Hz, 1H), 5.12 - 5.04 (m, 1H), 4.97 - 4.86 (m, 1H), 4.78 - 4.63 (m, 1H), 4.38 (t, J = 7.6 Hz, 1H), 4.35 - 4.27 (m, 2H), 4.07 - 3.98 (m, 2H), 3.80 - 3.69 (m, 1H), 3.61 (d, J = 11.2 Hz, 1H), 3.26 - 3.15 (m, 4H), 236 15077.006WO1 3.10 - 2.97 (m, 2H), 2.90 - 2.75 (m, 1H), 2.47 - 2.35 (m, 4H), 2.15 - 1.81 (m, 6H), 1.79 (s, 3H), 1.68 - 1.57 (m, 3H), 1.56 - 1.45 (m, 3H), 1.37 (d, J = 6.8 Hz, 3H), 1.02 (d, J = 6.0 Hz, 6H), 0.66 (d, J = 6.4 Hz, 3H). LCMS: MS (ESI) m/z 1034.5 [M+H]⁺. The following compounds were prepared in a manner similar for KV-13 using the 5 appropriate compound as starting material. 237 15077.006WO1 238 15077.006WO1 239 15077.006WO1 240 15077.006WO1 241 15077.006WO1 242 15077.006WO1 243 15077.006WO1 Example KV-45 Synthesis of N-(1,3-benzothiazol-2-yl)-2-[5-[1-[[3-(2- hydroxyethoxy)-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]-6-[[4-[[(1S)-1- [(2S,4R)-4-hydroxy-2-[[(1S)-1-[2-hydroxy-4-(4-methylthiazol-5-5 yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-4-oxo- butyl]sulfonylcarbamoyl]-2-pyridyl]-3,4-dihydro-1H-isoquinoline-8-carboxamide, KV-45 244 15077.006WO1 245 15077.006WO1 Step A: KV-45a To a solution of 3-(hydroxymethyl) cyclobutanone (1.30 g, 13.0 mmol, 1 eq) in DMF (13 mL) was added imidazole (1.33 g, 19.5 mmol, 1.5 eq) and tert-butyl(dimethyl)silyl chloride, 5 TBSCl (2.15 g, 14.3 mmol, 1.76 mL, 1.1 eq). The mixture was stirred at 25°C for 2 hrs. The mixture was poured into water 20 mL. The aqueous phase was extracted with EtOAc (10 mL*3). The combined organic phase was washed with brine (20 mL*3), dried with anhydrous Na2SO4, filtered and concentrated in vacuum. The residue was purified by column chromatography (SiO₂, Petroleum ether/Ethyl acetate=1/0 to 10/1) to give 3-[[tert-butyl(dimethyl)silyl] 10 oxymethyl] cyclobutanone, KV-45a (2.50 g, 11.7 mmol, 89.8% yield) was obtained as colorless oil.¹H NMR (400 MHz, DMSO-d6) δ 3.72 (d, J = 5.5 Hz, 2H), 3.10 - 2.99 (m, 2H), 2.80 - 2.70 (m, 2H), 2.58 - 2.51 (m, 1H), 0.86 (s, 9H), 0.05 (s, 6H) Step B: KV-45b To a solution of KV-45a (0.90 g, 4.20 mmol, 1 eq) in MeOH (9 mL) was added NaBH4 15 (238 mg, 6.30 mmol, 1.5 eq) at 0°C under N2 atmosphere. Then the mixture was slowly warmed to 25°C and stirred for 1 hr. The reaction mixture was quenched by addition NH₄Cl (15 mL) at 0°C. The mixture reaction was extracted with EtOAc (10 mL x 3). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO₂, 20 Petroleum ether/Ethyl acetate=1/0 to 5/1) to give 3-[[tert-butyl(dimethyl)silyl] oxymethyl] 246 15077.006WO1 cyclobutanol, KV-45b (1.00 g, crude) was obtained as colorless oil.¹H NMR (400 MHz, DMSO-d₆) δ 4.88 (d, J = 6.3 Hz, 1H), 3.96 - 3.85 (m, 1H), 3.55 - 3.47 (m, 2H), 2.18 - 2.09 (m, 2H), 1.92 - 1.73 (m, 1H), 1.55 - 1.44 (m, 2H), 0.86 (s, 9H), 0.03 -0.02 (m, 6H). Step C: KV-35c 5 To a solution of KV-45b (500 mg, 2.31 mmol, 1 eq) in THF (6 mL) was added NaH (138 mg, 3.47 mmol, 60% purity, 1.5 eq) at 0°C and stirred for 1 hr, then added 1,1,2- trichloroethylene (607 mg, 4.62 mmol, 607 μL, 2 eq) at 0°C. The reaction mixture was warmed to 25°C and stirred for 12 hrs. The reaction mixture was quenched by addition of ice-water 20 mL at 0 °C and then extracted with EtOAc (15 mL * 3). The combined organic layers were 10 washed with brine 20 mL, dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 12 g SepaFlash® Silica Flash Column, Eluent of 0~5% Ethyl acetate/Petroleum ethergradient @ 75 mL/min) to give tert-butyl-[[3-[(E)-1,2-dichlorovinyloxy]cyclobutyl]methoxy]-dimethyl-silane, KV-45c (300 mg, 963.64 μmol, 41.70% yield) was obtained as a colorless oil.¹H NMR (40015 MHz, DMSO-d6) δ 6.08 (s, 1H), 4.66-4.53 (m, 1H), 3.64-3.50 (m, 2H), 2.31-2.19 (m, 2H), 2.09- 2.02 (m, 1H), 2.01-1.91 (m, 2H), 0.88 (s, 9H), 0.031(s, 6H). Step D: KV-45d To a solution of KV-45c (200 mg, 642 μmol, 1 eq) in THF (5 mL) was added n-BuLi (2.5 M, 642 μL, 2.5 eq) dropwise at -78°C .The reaction mixture was warmed to -30°C and stirred 20 for 2 hrs. The reaction mixture was quenched by addition NH₄Cl.aq 20 mL at 0°C, and then extracted with EtOAc (15 mL * 3). The combined organic layers were washed with brine 20 mL, dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue, tert-butyl- [(3-ethynoxycyclobutyl) methoxy]-dimethyl-silane, KV-45d (150 mg, crude) as yellow oil was used into the next step without further purification. 25 Step E: KV-45e To a solution of KV-45d (150 mg, 305 μmol, 1 eq) in t-BuOH (2 mL), THF (2 mL) and H₂O (2 mL) were added tert-butyl-[(3-ethynoxycyclobutyl)methoxy]-dimethyl-silane (147 mg, 611 μmol, 2 eq), CuSO4 (24.4 mg, 152 μmol, 23.4 μL, 0.5 eq) and sodium;(2R)-2-[(1S)-1,2- dihydroxyethyl]-4-hydroxy-5-oxo-2H-furan-3-olate (90.8 mg, 458 μmol, 1.5 eq). The reaction 30 mixture was stirred at 30°C for 2 hrs. The reaction mixture was quenched by addition H₂O 10 mL at 25 °C, and then extracted with Ethyl acetate (10 mL * 3), the combined organic layers were washed with brine 15 mL, dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue, (2S,4R)-1-[(2S)-2-[4-[3-[[tert- butyl(dimethyl)silyl]oxymethyl]cyclobutoxy]triazol-1-yl]-3-methyl-butanoyl]-N-[(1S)-1-[5- 35 fluoro-2-hydroxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]-4-hydroxy-pyrrolidine-2-carboxamide, 247 15077.006WO1 KV-45e (220 mg, crude) as yellow oil was used into the next step without further purification. LCMS: MS (ESI) m/z 731.30 [M+H]⁺. Step F: KV-45f To a solution of KV-45e (220 mg, 300 μmol, 1 eq) in DCM (5 mL) were added TEA 5 (60.9 mg, 601 μmol, 83.7 μL, 2 eq) and DMAP (3.68 mg, 30.1 μmol, 0.1 eq), Ac2O (76.8 mg, 752 μmol, 70.6 μL, 2.5 eq). The reaction mixture was stirred at 25°C for 1 hr. The reaction mixture was diluted with ice-water 10 mL and extracted with DCM (10 mL * 3). The combined organic layers were washed with brine 10 mL, dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue, [2-[(1S)-1-[[(2S,4R)-4-acetoxy-1-[(2S)-2-[4-[3-[[tert-10 butyl(dimethyl)silyl]oxymethyl]cyclobutoxy]triazol-1-yl]-3-methyl-butanoyl]pyrrolidine-2- carbonyl]amino]ethyl]-4-fluoro-5-(4-methylthiazol-5-yl)phenyl] acetate, KV-45f (250 mg, crude) as yellow oil was used into the next step without further purification. LCMS: MS (ESI) m/z 815.30 [M+H]⁺. Step G: KV-45g 15 To a solution of KV-45f (250 mg, 306 μmol, 1 eq) in MeOH (5 mL) was added acetyl chloride (36.1 mg, 460 μmol, 32.7 μL, 1.5 eq). The reaction mixture was stirred at 25°C for 1 hr. The reaction mixture was added NaHCO3.aq to pH≈8 at 0 °C, and then extracted with EtOAc (10 mL * 3). The combined organic layers were washed with brine 10 mL, dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue, [2-[(1S)-1-[[(2S,4R)-4-20 acetoxy-1-[(2S)-2-[4-[3-(hydroxymethyl)cyclobutoxy]triazol-1-yl]-3-methyl- butanoyl]pyrrolidine-2-carbonyl]amino]ethyl]-4-fluoro-5-(4-methylthiazol-5-yl)phenyl] acetate, KV-45g (220 mg, crude) as a yellow oil was used into the next step without further purification. LCMS: MS (ESI) m/z 701.20 [M+H]⁺. Step H: KV-45h 25 To a solution of KV-45g (200 mg, 285 μmol, 1 eq) in DCM (5 mL) was added Dess- Martin (181 mg, 428 μmol, 132 μL, 1.5 eq). The reaction mixture was stirred at 25°C for 1 hr. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 4 g SepaFlash® Silica Flash Column, Eluent of 0~100% Ethyl acetate/Petroleum ethergradient @ 75 mL/min) to give30 [2-[(1S)-1-[[(2S,4R)-4-acetoxy-1-[(2S)-2-[4-(3-formylcyclobutoxy)triazol-1-yl]-3-methyl- butanoyl]pyrrolidine-2-carbonyl]amino]ethyl]-4-fluoro-5-(4-methylthiazol-5-yl)phenyl] acetate, KV-45h (130 mg, 186 μmol, 65.19% yield) was obtained as a yellow oil. LCMS: MS (ESI) m/z 699.30 [M+H]⁺. Step I: KV-45i 248 15077.006WO1 To a solution of KV-45h (50.0 mg, 71.5 μmol, 1 eq) in DCM (3 mL) was added NaBH(OAc)₃ (60.6 mg, 286 μmol, 4 eq) and (4S)-2-amino-4-methyl-4-[3-[2-[(2S)-2-methyl- 1,4-diazepan-1-yl]pyrimidin-4-yl]-1,2,4-oxadiazol-5-yl]-6,7-dihydro-5H-benzothiophene-3- carbonitrile (25.8 mg, 57.2 μmol, 0.8 eq). The reaction mixture was stirred at 25°C for 1 hr. The 5 reaction mixture was diluted with ice-water 10 mL and extracted with DCM (10 mL *3). The combined organic layers were washed with brine 10 mL, dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue, [2-[(1S)-1-[[(2S,4R)-4-acetoxy-1-[(2S)- 2-[4-[3-[[(3S)-4-[4-[5-[(4S)-2-amino-3-cyano-4-methyl-6,7-dihydro-5H-benzothiophen-4-yl]- 1,2,4-oxadiazol-3-yl]pyrimidin-2-yl]-3-methyl-1,4-diazepan-1-yl]methyl]cyclobutoxy]triazol-1-10 yl]-3-methyl-butanoyl]pyrrolidine-2-carbonyl]amino]ethyl]-4-fluoro-5-(4-methylthiazol-5- yl)phenyl] acetate, KV-45i (95 mg, crude) as yellow oil was used into the next step without further purification. LCMS: MS (ESI) m/z 1133.30 [M+H]⁺. Step J: KV-45 To a solution of KV-45i (90.0 mg, 79.4 μmol, 1 eq) in MeOH (2 mL) was added K2CO3 15 (54.8 mg, 397 μmol, 5 eq). The reaction mixture was stirred at 20°C for 1 hr. The reaction mixture was added TFA to pH≈5 and concentrated under reduced pressure to remove MeOH. The residue was purified by prep-HPLC (column: Phenomenex Luna C18 75*30mm*3um;mobile phase: [H₂O(0.1% TFA)-ACN];gradient:15%-45% B over 8.0 min) to give KV-45 (30.0 mg, 28.2 μmol, 35.5% yield, 98.80% purity) as a white solid.¹H NMR (400 20 MHz, DMSO-d6) δ 9.80 (s, 1H), 9.50-9.46 (m, 1H), 9.08 (s, 1H), 8.72-8.60 (m, 1H), 8.42 (d, J = 8.0 Hz, 1H), 7.71-7.58 (m, 1H), 7.33-7.23 (m, 1H), 7.12-7.02 (m, 2H), 6.90-6.76 (m, 1H), 5.19- 5.05 (m, 2H), 5.00-4.79 (m, 1H), 4.72-4.57 (m, 1H), 4.48-4.25 (m, 3H), 3.78-3.71 (m, 1H), 3.67- 3.51 (m, 4H), 3.40-3.35 (m, 4H), 3.22-3.13 (m, 2H), 2.50-2.41 (m, 4H), 2.36-2.33 (m, 3H), 2.15- 2.03 (m, 2H), 2.01-1.80 (m, 8H), 1.79 (s, 3H), 1.31 (d, J = 6.8 Hz, 3H), 1.20-1.09 (m, 3H), 1.04 25 (d, J = 6.8 Hz, 3H), 0.66 (d, J = 6.4 Hz, 3H). LCMS: MS (ESI) m/z 1049.40 [M+H]⁺. The following compounds were prepared in a manner similar to KV-45 using the appropriate compound as starting material. 249 15077.006WO1 250 15077.006WO1 Example KV-53 Synthesis of (2S,4R)-1-[(2S)-2-[4-[4-[4-[[1-[[4-[5-[(4S)-2-amino- 3-cyano-4-methyl-6,7-dihydro-5H-benzothiophen-4-yl]-1,2,4-oxadiazol-3-yl]pyrimidin-2- yl]oxymethyl]cyclopropyl]methyl]piperazin-1-yl]butoxy]triazol-1-yl]-3-methyl-butanoyl]-N- 5 [(1S)-1-[3-fluoro-2-hydroxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]-4-hydroxy-pyrrolidine-2- carboxamide, KV-53 10 Step 1: Preparation of [1-[[tert-butyl(dimethyl)silyl]oxymethyl]cyclopropyl]methanol, KV-53a To a solution of [1-(hydroxymethyl)cyclopropyl]methanol (20 g, 195 mmol, 1 eq) in DCM (2000 mL) was added TBSCl (30.9 g, 205 mmol, 25.3 mL, 1.05 eq) and imidazole (20.0 g, 293 mmol, 1.5 eq) in one portion at 0°C. The reaction mixture was warmed to 20°C and 15 stirred for 2 hrs. The reaction mixture was diluted with H2O (500 mL) and extracted with DCM (200 mL x 3). The combined organic layers was washed with brine (200 mL), dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to dryness. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 100/1 to 5/1). to give [1-[[tert-butyl (dimethyl) silyl] oxymethyl] cyclopropyl] methanol, KV-53a (20 g, 92.4 20 mmol, 47.2% yield) as a colorless oil.¹H NMR: (400 MHz, CDCl3) δ3.60 (s, 2H), 3.55 (s, 2H), 2.89 - 2.49 (m, 1H), 0.90 (s, 9H), 0.54 - 0.46 (m, 2H), 0.46 - 0.39 (m, 2H), 0.06 (s, 6H) 251 15077.006WO1 Step 2: Preparation of 2-[[1-[[tert-butyl (dimethyl) silyl] oxymethyl] cyclopropyl] methoxy] pyrimidine-4-carbonitrile, KV-53b To a mixture of 2-chloropyrimidine-4-carbonitrile (10.0 g, 71.6 mmol, 1 eq) and KV-43a (18.6 g, 86.0 mmol, 1.2 eq) in toluene (400 mL) was added Cs2CO3 (35.0 g, 107 mmol, 1.5 eq) 5 and [2-(2-aminophenyl)phenyl]-methylsulfonyloxy-palladium;ditert-butyl-[3,6-dimethoxy-2- (2,4,6-triisopropylphenyl)phenyl]phosphane (1.22 g, 1.43 mmol, 0.02 eq) under N₂. The reaction mixture was stirred and heated at 80°C for 16 hrs. After filtration via celite pad, the organic layer was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 100/1 to 5/1) to give 2-[[1-[[tert- 10 butyl(dimethyl)silyl]oxymethyl]cyclopropyl]methoxy]pyrimidine-4-carbonitrile, KV-53b (4 g, 11.2 mmol, 15.7% yield, 90% purity) as a yellow oil.¹ H NMR (400 MHz, CDCl3) δ8.72 (d, J = 4.8 Hz, 1H), 7.25 (d, J = 4.8 Hz, 1H), 4.34 (s, 2H), 3.64 (s, 2H), 0.90 - 0.81 (m, 9H), 0.60 (s, 4H), 0.01 (s, 6H). LCMS: MS (ESI) m/z 320.2 [M+H]⁺ Step 3: Preparation of 2-[[1-[[tert- 15 butyl(dimethyl)silyl]oxymethyl]cyclopropyl]methoxy]-N-hydroxy-pyrimidine-4-carboxamidine, KV-53c To a solution of KV-53b (3.80 g, 11.8 mmol, 1 eq) in EtOH (30 mL) was added hydroxylamine (3.14 g, 47.5 mmol, 50% purity, 4 eq). The reaction mixture was stirred and heated at 60°C for 2 hrs. The reaction mixture was concentrated under reduced pressure to 20 dryness. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 100/1 to 5/1) to give KV-53c (3.8 g, 10.7 mmol, 90.6% yield) as a yellow oil.¹H NMR: (400 MHz, CDCl₃) δ = 8.49 (d, J = 5.2 Hz, 1H), 7.45 (d, J = 5.2 Hz, 1H), 5.55 (s, 2H), 4.34 (s, 2H), 3.68 (s, 2H), 0.89 - 0.85 (m, 9H), 0.62 (s, 4H), 0.04 - -0.03 (m, 6H). LCMS: MS (ESI) m/z 353.2 [M+H]⁺ 25 Step 4: Preparation of (NZ,4S)-2-amino-N-[[2-[[1-[[tert- butyl(dimethyl)silyl]oxymethyl]cyclopropyl]methoxy]pyrimidin-4-yl]- (hydroxyamino)methylene]-3-cyano-4-methyl-6,7-dihydro-5H-benzothiophene-4-carboxamide, KV-53d To a solution of (4S)-2-amino-3-cyano-4-methyl-6, 7-dihydro-5H-benzothiophene-4- 30 carboxylic acid (2.01 g, 8.51 mmol, 1 eq) in DMSO (50 mL) was added HATU (3.56 g, 9.36 mmol, 1.1 eq) and TEA (2.58 g, 25.5 mmol, 3.55 mL, 3 eq) in one portion. The reaction mixture was stirred at 25°C for 0.5 hr. Then KV-43c (3 g, 8.51 mmol, 1 eq) was added to the reaction mixture. The reaction mixture was stirred at 25°C for 16 hrs. The reaction mixture was diluted with H2O (30 mL) and extracted with EtOAc (30 mL x 3). The combined organic layers was 35 washed with brine (30 mL), dried over anhydrous Na2SO4, filtered and concentrated under 252 15077.006WO1 reduced pressure to dryness. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 100/1 to 1/1) to give KV-53d (3.7 g, 6.48 mmol, 76.1% yield) as a yellow oil.¹H NMR: (400 MHz, CDCl₃-d₆) δ = 8.55 (d, J = 5.2 Hz, 1H), 7.72 (d, J = 5.2 Hz, 1H), 4.77 (s, 1H), 4.33 (s, 2H), 3.66 (s, 2H), 2.66 - 2.40 (m, 3H), 2.04 - 1.88 (m, 2H), 1.78 (s, 5 3H), 1.67 - 1.56 (m, 1H), 0.87 (s, 9H), 0.61 (s, 4H), 0.01 (s, 6H). LCMS: MS (ESI) m/z 571.2 [M+H]⁺ Step 5: Preparation of (4S)-2-amino-4-[3-[2-[[1-[[tert- butyl(dimethyl)silyl]oxymethyl]cyclopropyl]methoxy]pyrimidin-4-yl]-1,2,4-oxadiazol-5-yl]-4- methyl-6,7-dihydro-5H-benzothiophene-3-carbonitrile, KV-53e 10 To a solution of KV-53d (3.8 g, 6.66 mmol, 1 eq) in THF (40 mL) was added DBU (2.53 g, 16.6 mmol, 2.51 mL, 2.5 eq). The reaction mixture was stirred and heated at 60°C for 12 hrs. After cooling to room temperature, the reaction mixture was diluted with H₂O (30 mL) and extracted with EtOAc (20 mL x 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to dryness. 15 The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 100/1 to 1/1) to give KV-53e (3 g, 5.43 mmol, 81.5% yield) as a yellow oil.¹H NMR: (400 MHz, CDCl3-d6) δ8.68 (d, J = 4.8 Hz, 1H), 7.68 (d, J = 4.8 Hz, 1H), 4.78 (s, 2H), 4.39 (s, 2H), 3.69 (s, 2H), 2.70 - 2.57 (m, 2H), 2.41 - 2.29 (m, 1H), 2.02 - 1.84 (m, 5H), 1.79 - 1.56 (m, 2H), 0.87 (s, 9H), 0.66 - 0.55 (m, 4H), 0.01 (s, 6H). LCMS: MS (ESI) m/z 553.2 [M+H]⁺ 20 Step 6: Preparation of tert-butyl N-[(4S)-4-[3-[2-[[1-[[tert-butyl (dimethyl)silyl] oxymethyl] cyclopropyl] methoxy] pyrimidin-4-yl]-1,2,4-oxadiazol-5-yl]-3-cyano-4-methyl-6,7- dihydro-5H-benzothiophen-2-yl]carbamate, KV-53f To a solution of KV-53e (0.5 g, 904 μmol, 1 eq) in MeCN (5 mL) was added DMAP (221 mg, 1.81 mmol, 2 eq) and tert-butyl (4-nitrophenyl) carbonate (432 mg, 1.81 mmol, 2 eq) and 25 K2CO3 (1.00 g, 7.24 mmol, 8 eq). The reaction mixture was stirred and heated at 60°C for 4 hrs. After cooling to room temperature, the reaction mixture was diluted with H₂O (20 mL) and extracted with EtOAc (20 mL x 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to dryness. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 30 100/1 to 3/1) to give KV-53f (0.56 g, 857 μmol, 94.8% yield) as a yellow solid. LCMS: MS (ESI) m/z 653.3 [M+H]⁺ Step 7: Preparation of tert-butyl N-[(4S)-3-cyano-4-[3-[2-[[1-(hydroxymethyl) cyclopropyl] methoxy] pyrimidin-4-yl]-1, 2, 4-oxadiazol-5-yl]-4-methyl-6,7-dihydro-5H- benzothiophen-2-yl]carbamate, KV-53g 253 15077.006WO1 To a solution of KV-53f (0.56 g, 857 μmol, 1 eq) in DMF (5 mL) was added CsF (651 mg, 4.29 mmol, 5 eq). The reaction mixture was stirred at 70°C for 2 hrs. After cooling to room temperature, the reaction mixture was diluted with H2O (20 mL) and extracted with EtOAc (20 5 mL x 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to dryness to give KV-53g (0.46 g, crude) as a yellow solid. LCMS: MS (ESI) m/z 539.2 [M+H]⁺ Step 8: Preparation of tert-butyl N-[(4S)-3-cyano-4-[3-[2-[(1-formylcyclopropyl) methoxy] pyrimidin-4-yl]-1, 2, 4-oxadiazol-5-yl]-4-methyl-6, 7-dihydro-5H-benzothiophen-2- 10 yl] carbamate, KV-53h To a solution of KV-53g (0.46 g, 854 μmol, 1 eq) in DCM (5 mL) was added Dess- Martin reagent (543 mg, 1.28 mmol, 1.5 eq). The reaction mixture was stirred at 25°C for 1 hr. The mixture was adjusted to pH = 7 with saturated aqueous NaHCO3 and extracted with DCM (10 mL x 3). The combined organic phase was washed with brine (10 mL), dried with 15 anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to dryness. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 100/1 to 1/1) to give KV-53h (0.27 g, 503 μmol, 58.9% yield) as a yellow solid.¹H NMR: (400 MHz, CDCl3- d6) δ9.26 (s, 1H), 8.70 (d, J = 4.8 Hz, 1H), 7.71 (d, J = 4.8 Hz, 1H), 7.54 (s, 1H), 4.73 (s, 2H), 2.82 - 2.64 (m, 2H), 2.43 - 2.30 (m, 1H), 2.04 - 1.88 (m, 7H), 1.71 - 1.58 (m, 3H), 1.40 - 1.35 20 (m, 2H), 1.34 - 1.29 (m, 2H). LCMS: MS (ESI) m/z 537.2 [M+H]⁺ Step 9: Preparation of tert-butyl 4-[4-[1-[(1S)-1-[(2S,4R)-4-acetoxy-2-[[(1S)-1-[2- acetoxy-3-fluoro-4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2- methyl-propyl]triazol-4-yl]oxybutyl]piperazine-1-carboxylate, KV-53i 254 15077.006WO1 5 To a solution of tert-butyl piperazine-1-carboxylate (25.9 mg, 116 μmol, 2 eq, HCl) in DCM (1 mL) was added TEA until pH =10, then AcOH was added and adjusted pH to 7. Then added [6-[(1S)-1-[[(2S,4R)-4-acetoxy-1-[(2S)-3-methyl-2-[4-(4-oxobutoxy)triazol-1-yl] butanoyl]pyrrolidine-2-carbonyl]amino]ethyl]-2-fluoro-3-(4-methylthiazol-5-yl)phenyl] acetate 255 15077.006WO1 (40 mg, 58.2 μmol, 1 eq) and NaBH(OAc)3 (61.7 mg, 291 μmol, 5 eq). The reaction mixture was stirred at 25°C for 2 hrs. The reaction mixture was diluted with H₂O (10 mL) and extracted with DCM (5 mL x 3). The combined organic layers were washed with brine (5 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to dryness. The residue 5 was purified by prep-TLC (SiO2, EtOAc:MeOH = 10:1) to give KV-53i (46 mg, 53.6 μmol, 92.1% yield) was obtained as a yellow solid. LCMS: MS (ESI) m/z 857.3 [M+H]⁺ Step 10: Preparation of [6-[(1S)-1-[[(2S,4R)-4-acetoxy-1-[(2S)-3-methyl-2-[4-(4- piperazin-1-ylbutoxy)triazol-1-yl]butanoyl]pyrrolidine-2-carbonyl]amino]ethyl]-2-fluoro-3-(4- methylthiazol-5-yl)phenyl] acetate, KV-53j 10 To a solution of KV-53i (40 mg, 46.6 μmol, 1 eq) in EtOAc (1 mL) was added HCl/EtOAc (4 M, 2.40 mL, 205 eq). The reaction mixture was stirred at 25°C for 2 hrs. The reaction mixture was concentrated under reduced pressure to dryness. Compound [6-[(1S)-1- [[(2S,4R)-4-acetoxy-1-[(2S)-3-methyl-2-[4-(4-piperazin-1-ylbutoxy)triazol-1- yl]butanoyl]pyrrolidine-2-carbonyl]amino]ethyl]-2-fluoro-3-(4-methylthiazol-5-yl)phenyl] 15 acetate, KV-53j (40 mg, crude, HCl) was obtained as a yellow solid. LCMS: MS (ESI) m/z 757.3 [M+H]⁺ Step 11: Preparation of [6-[(1S)-1-[[(2S,4R)-4-acetoxy-1-[(2S)-2-[4-[4-[4-[[1-[[4-[5- [(4S)-2-(tert-butoxycarbonylamino)-3-cyano-4-methyl-6,7-dihydro-5H-benzothiophen-4-yl]- 1,2,4-oxadiazol-3-yl]pyrimidin-2-yl]oxymethyl]cyclopropyl]methyl]piperazin-1-20 yl]butoxy]triazol-1-yl]-3-methyl-butanoyl]pyrrolidine-2-carbonyl]amino]ethyl]-2-fluoro-3-(4- methylthiazol-5-yl)phenyl] acetate, KV-53k To a solution of KV-53j (39.9 mg, 50.3 μmol, 0.9 eq, HCl) in MeOH (1 mL) was added NaOAc (45.8 mg, 559 μmol, 10 eq). The reaction mixture was stirred at 25°C for 0.5 h. Then added NaBH3CN (7.03 mg, 111 μmol, 2 eq) and tert-butyl N-[(4S)-3-cyano-4-[3-[2-[(1-25 formylcyclopropyl)methoxy]pyrimidin-4-yl]-1,2,4-oxadiazol-5-yl]-4-methyl-6,7-dihydro-5H- benzothiophen-2-yl]carbamate, KV-43h (30 mg, 55.9 μmol, 1 eq). The reaction mixture was stirred and heated at 40°C for 16 hrs. The reaction mixture was diluted with H₂O (10 mL) and extracted with DCM (5 mL x 3). The combined organic layers were washed with brine (5 mL), dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to dryness to 30 give KV-53k (65 mg, crude) was obtained as a white solid. LCMS: MS (ESI) m/z 1277.4 [M+H]⁺ Step 12: Preparation of tert-butyl N-[(4S)-3-cyano-4-[3-[2-[[1-[[4-[4-[1-[(1S)-1-[(2S,4R)- 2-[[(1S)-1-[3-fluoro-2-hydroxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]-4-hydroxy- pyrrolidine-1-carbonyl]-2-methyl-propyl]triazol-4-yl]oxybutyl]piperazin-1- 256 15077.006WO1 yl]methyl]cyclopropyl]methoxy]pyrimidin-4-yl]-1,2,4-oxadiazol-5-yl]-4-methyl-6,7-dihydro- 5H-benzothiophen-2-yl]carbamate, KV-53l To a solution of KV-53k (50 mg, 39.1 μmol, 1 eq) in MeOH (0.5 mL) was added K₂CO₃ (10.8 mg, 78.2 μmol, 2 eq). The reaction mixture was stirred at 25°C for 2 hrs. The mixture 5 was adjusted to pH = 7 with TFA and extracted with DCM (5 mL x 3). The combined organic phase was washed with brine (5 mL), dried with anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to dryness to give KV-53l (40 mg, crude) as a white solid. LCMS: MS (ESI) m/z 1193.4 [M+H]⁺ Step 13: Preparation of (2S,4R)-1-[(2S)-2-[4-[4-[4-[[1-[[4-[5-[(4S)-2-amino-3-cyano-4-10 methyl-6,7-dihydro-5H-benzothiophen-4-yl]-1,2,4-oxadiazol-3-yl]pyrimidin-2- yl]oxymethyl]cyclopropyl]methyl]piperazin-1-yl]butoxy]triazol-1-yl]-3-methyl-butanoyl]-N- [(1S)-1-[3-fluoro-2-hydroxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]-4-hydroxy-pyrrolidine-2- carboxamide, KV-53 To a solution of KV-53l (40 mg, 33.5 μmol, 1 eq) in EtOAc (0.5 mL) was added 15 HCl/EtOAc (4 M, 3 mL, 358 eq). The reaction mixture was stirred at 25°C for 2 hrs. The reaction mixture was concentrated under reduced pressure to dryness. The residue was purified by prep-HPLC (column: WePure Biotech XP tC18150 x 40 x 70um; mobile phase: [H2O (10mM NH4HCO3)-ACN]; gradient: 25%-70% B over 8.0 min) to give KV-53 (2.6 mg, 2.25 μmol, 6.70% yield, 94.5% purity) as a white solid.1H NMR (400 MHz, DMSO-d₆) δ9.07 (s, 20 1H), 8.82 (d, J = 4.8 Hz, 1H), 8.63 - 8.45 (m, 1H), 7.75 - 7.58 (m, 2H), 7.08 (s, 2H), 7.04 - 6.78 (m, 1H), 5.23 - 5.00 (m, 3H), 4.45 - 4.35 (m, 2H), 4.30 (s, 4H), 2.45 - 2.16 (m, 18H), 2.13 - 2.01 (m, 4H), 1.91 - 1.75 (m, 6H), 1.73 - 1.39 (m, 6H), 1.33 (d, J = 6.4 Hz, 3H), 1.07 - 0.96 (m, 3H), 0.66 (d, J = 6.0 Hz, 3H), 0.43-0.40 (m, 2H). LCMS: MS (ESI) m/z 1093.4[M+H]⁺ Example BXV-41 Synthesis of N-(1,3-benzothiazol-2-yl)-2-[5-[1-[[3-(2-25 hydroxyethoxy)-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]-6-[[4-[[(1S)-1- [(2S,4R)-4-hydroxy-2-[[(1S)-1-[2-hydroxy-4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-4-oxo- butyl]sulfonylcarbamoyl]-2-pyridyl]-3,4-dihydro-1H-isoquinoline-8-carboxamide, BXV-41 Step A. Preparation of Int 8a 257 15077.006WO1 To a solution of 2-[[3-[(4-iodo-5-methyl-pyrazol-1-yl)methyl]-5,7-dimethyl-1- adamantyl]oxy]ethanol (1 g, 2.25 mmol, 1 eq) and tert-butyl 6-[8-(1,3-benzothiazol-2- ylcarbamoyl)-3,4-dihydro-1H-isoquinolin-2-yl]-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2- 5 yl)pyridine-2-carboxylate (1.65 g, 2.70 mmol, 1.2 eq) in dioxane (10 mL) and H2O (2 mL) were added K2CO3 (622.06 mg, 4.50 mmol, 2 eq) and [2-(2- aminophenyl)phenyl]palladium(1+);bis(1-adamantyl)-butyl-phosphane;methanesulfonate (163.90 mg, 225.05 μmol, 0.1 eq) at 20°C under N₂. The system was degassed and then charged with N₂ for three times. The reaction mixture was heated to 90 °C and stirred for 3 hrs. After 10 cooling to room temperature, the reaction mixture was poured into water (20 mL) , extracted with EtOAc (15 mL x 3). The combined organic layers were washed with brine (10 mL x 1), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 12 g SepaFlash® Silica Flash Column, Eluent of 0~70% ethyl acetate/petroleum ether gradient @ 100mL/min) to give tert-15 butyl 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydro-1H-isoquinolin-2-yl]-3-[1-[[3-(2- hydroxyethoxy)-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2- carboxylate Int 8a (1.45 g, 1.72 mmol, 76.40% yield, 95.22% purity) as a yellow solid.¹ H NMR (400 MHz, DMSO-d6) δ12.86 (s, 1H), 8.02 (d, J = 7.6 Hz, 1H), 7.78 (d, J = 7.6 Hz, 1H), 7.58 (d, J = 7.6 Hz, 1H), 7.50 - 7.41 (m, 3H), 7.39 - 7.31 (m, 2H), 7.21 (s, 1H), 6.92 (d, J = 8.8 20 Hz, 1H), 4.96 (s, 2H), 4.44 (t, J = 5.6 Hz, 1H), 3.87 - 3.79 (m, 4H), 3.41 - 3.35 (m, 3H), 3.32 - 3.29 (m, 4H), 3.02 (t, J = 6.0 Hz, 2H), 2.08 (s, 3H), 1.38-0.92 (m, 21H), 0.82 (s, 6H). LCMS: MS (ESI) m/z 803.4 [M+H]⁺ Step B. Preparation of Int 8b 258 15077.006WO1 To a solution of Int 8a (1.4 g, 1.74 mmol, 1 eq) in DCM (20 mL) was added TFA (4 mL) at 20 °C. The reaction mixture was heated to 30°C and stirred for 16 hrs. The reaction mixture was concentrated in vacuum to give a residue. The residue was triturated with hexane 5 (50 mL) at 20oC for 30 min. Compound 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydro-1H- isoquinolin-2-yl]-3-[1-[[3,5-dimethyl-7-[2-(2,2,2-trifluoroacetyl) oxyethoxy]-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carboxylic acid Int 8b (1.9 g, 1.46 mmol, 83.49% yield, 73.31% purity, TFA) was obtained as a yellow solid. LCMS: MS (ESI) m/z 843.2 [M+H]⁺ 10 Step C. Preparation of Int 8c To a solution of Int 8b (1.7 g, 1.78 mmol, 1 eq, TFA) in MeOH (17 mL) was added K2CO3 (737 mg, 5.33 mmol, 3 eq) at 20°C and stirred for 1 h. The reaction mixture was poured into water (50 mL), extracted with EtOAc (20 mL x 3). The combined organic layers were 15 washed with brine (10 mL), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was used in the next step directly without further purification. Compound 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydro-1H-isoquinolin-2-yl]-3-[1-[[3- (2-hydroxyethoxy)-5,7-dimethyl-1-adamantyl] methyl]-5-methyl-pyrazol-4-yl]pyridine-2- carboxylic acid Int 8c (1.2 g, 1.47 mmol, 82.77% yield, 91.52% purity) was obtained as a 20 yellow solid.¹H NMR: (400 MHz, DMSO-d6) δ 8.03 (d, J = 7.6 Hz, 1H), 7.79 (d, J = 8.0 Hz, 1H), 7.62 (d, J = 7.2 Hz, 1H), 7.52 - 7.40 (m, 3H), 7.39 - 7.32 (m, 2H), 7.29 (s, 1H), 6.93 (d, J = 8.8 Hz, 1H), 4.95 (s, 2H), 3.88 (t, J = 5.6 Hz, 2H), 3.81 (s, 2H), 3.38 – 3.30 (m, 4H), 3.01 (t, J = 5.2 Hz, 2H), 2.10 (s, 3H), 1.36-0.94 (m, 12H), 0.90 - 0.80 (m, 6H) LCMS: MS (ESI) m/z 747.3 [M+H]⁺ 25 Step D. Preparation of Int 8d 259 15077.006WO1 To a solution of Int 8c (1.15 g, 1.54 mmol, 1 eq) in DCM (24 mL) were added imidazole (524.08 mg, 7.70 mmol, 5 eq) and TBSCl (348.09 mg, 2.31 mmol, 284.16 μL, 1.5 eq) at 0°C. The reaction mixture was warmed to 20°C and stirred for 2 hrs. The reaction mixture was 5 poured into water (40 mL), extracted with DCM (20 mL x 3). The combined organic layers were washed with brine (10 mL x 1), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was triturated with n-heptane at 20 ^oC for 20 min. Compound 6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydro-1H-isoquinolin-2-yl]-3-[1-[[3- [2-[tert-butyl(dimethyl)silyl]oxyethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4- 10 yl]pyridine-2-carboxylic acid Int 8d (1.17 g, 1.16 mmol, 75.08% yield, 85.09% purity) was obtained as a white solid. LCMS: MS (ESI) m/z 861.3 [M+H]⁺ Step E. Preparation of Int 8e To a solution of Int 8d (0.8 g, 929 μmol, 1 eq) and methyl 4-sulfamoylbutanoate (337 15 mg, 1.86 mmol, 2 eq) in DCM (8 mL) were added DMAP (453.95 mg, 3.72 mmol, 4 eq) and EDCI (534 mg, 2.79 mmol, 3 eq) at 0°C. The reaction mixture was warmed slowly to 20°C and stirred for 16 hrs. The reaction mixture was poured into water (20 mL), extracted with DCM (10 mL x 3). The combined organic layers were washed with brine (5 mL x 1), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by 20 flash silica gel chromatography (ISCO®; 12 g SepaFlash® Silica Flash Column, Eluent of 0~100% Ethyl acetate/Petroleum ethergradient @ 100 mL/min) to give methyl 4-[[6-[8-(1,3- benzothiazol-2-ylcarbamoyl)-3,4-dihydro-1H-isoquinolin-2-yl]-3-[1-[[3-[2-[tert- butyl(dimethyl)silyl]oxyethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4- yl]pyridine-2-carbonyl]sulfamoyl]butanoate Int 8e (0.5 g, 414.89 μmol, 44.66% yield, 85% 25 purity) as a white solid.¹ H NMR:(400 MHz, DMSO-d6) δ 8.13 (d, J = 6.8 Hz, 2H), 8.02 (d, J = 260 15077.006WO1 7.6 Hz, 1H), 7.78 (d, J = 8.0 Hz, 1H), 7.61 (d, J = 7.6 Hz, 1H), 7.51 - 7.29 (m, 6H), 6.80 (d, J = 9.2 Hz, 1H), 6.76 - 6.70 (m, 2H), 4.89 (s, 2H), 3.92 (t, J = 5.2 Hz, 2H), 3.78 (s, 2H), 3.61 - 3.54 (m, 5H), 3.35 - 3.30 (m, 2H), 3.11 - 2.96 (m, 4H), 2.42 - 2.36 (m, 2H), 2.12 (s, 3H), 1.80 - 1.71 (m, 2H), 1.17 - 1.09 (m, 4H), 1.40 - 0.95 (m, 12H), 0.83 (s, 15H), 0.01 (s, 6H) LCMS: MS (ESI) 5 m/z 1024.4 [M+H]⁺ Step F. Preparation of Int 8f To a solution of Int 8e (0.48 g, 398 μmol, 1 eq) in MeOH (5 mL) , THF (5 mL) and H2O (5 mL) was added LiOH.H2O (83.5 mg, 1.99 mmol, 5 eq) at 20°C. The reaction mixture was 10 stirred at 20°C for 1 h. The reaction mixture was poured into water (20 mL), extracted with MTBE (10 mL x 3). The combined organic layers were discarded. The aqueous phase was cooled to 0°C, acidified with 0.5 N HCl to pH=6~7, extracted with EtOAc(10 mL x 5). The combined organic layers were washed with brine (5 mL x 2), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was used into the next step15 directly without further purification. Compound 4-[[6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4- dihydro-1H-isoquinolin-2-yl]-3-[1-[[3-[2-[tert-butyl(dimethyl)silyl] oxyethoxy]-5,7-dimethyl-1- adamantyl]methyl]-5-methyl-pyrazol-4-yl]pyridine-2-carbonyl]sulfamoyl]butanoic acid Int 8f (0.38 g, 347.68 μmol, 87.29% yield, 92.44% purity) was obtained as a white solid.¹ H NMR (400 MHz, DMSO-d₆) δ = 8.02 (d, J = 8.0 Hz, 1H), 7.79 (d, J = 8.0 Hz, 1H), 7.60 (d, J = 7.6 Hz, 20 1H), 7.51 - 7.39 (m, 3H), 7.38 - 7.28 (m, 3H), 6.69 (d, J = 7.6 Hz, 1H), 4.84 (s, 2H), 3.92 (t, J = 5.6 Hz, 2H), 3.78 (s, 2H), 3.59 (t, J = 5.2 Hz, 2H), 2.99 – 2.93 (m, 4H), 2.61 - 2.57 (m, 2H), 2.29-2.26 (m, 2H), 2.14 (s, 3H), 1.74 - 1.66 (m, 2H), 1.35-0.96 (m, 12H), 0.89 - 0.76 (m, 15H), 0.01 (s, 6H) LCMS: MS (ESI) m/z 1010.3 [M+H]⁺ Step G. Preparation of Int 8g 261 15077.006WO1 To a solution of Int 8f (67.0 mg, 66.3 μmol, 1.1 eq) in DMF (0.5 mL) was added HATU (25.2 mg, 66.3 μmol, 1.1 eq) and DIEA (23.4 mg, 181 μmol, 31.5 μL, 3 eq) and (2S,4R)-1-[(2S)- 2-amino-3,3-dimethyl-butanoyl]-4-hydroxy-N-[(1S)-1-[2-hydroxy-4-(4-methylthiazol-5- 5 yl)phenyl]ethyl]pyrrolidine-2-carboxamide (0.03 g, 60.3 μmol, 1 eq, HCl). The reaction mixture was stirred at 20°C for 1 hr. The reaction mixture was diluted with H2O (10 mL) and extracted with DCM (5 mL x 3). The combined organic layers was washed with brine (5 mL), dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to dryness. The crude product was used for the next step without further purification. Compound N-(1,3-benzothiazol-10 2-yl)-2-[5-[1-[[3-[2-[tert-butyl(dimethyl)silyl]oxyethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5- methyl-pyrazol-4-yl]-6-[[4-[[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[2-hydroxy-4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]amino]-4-oxo-butyl]sulfonylcarbamoyl]-2-pyridyl]-3,4-dihydro-1H-isoquinoline-8- carboxamide Int 8g (0.07 g, crude) was obtained as a yellow solid. LCMS: MS (ESI) m/z 15 1452.5 [M+H]⁺ Step H. Preparation of BXV-41 262 15077.006WO1 To a solution of Int 8g (0.05 g, 34.4 μmol, 1 eq) in DCM (0.5 mL) TFA (2 mL) was added TFA (2 mL). The reaction mixture was stirred at 20°C for 1 hr. The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (column: WePure Biotech XP tC18150 x 40 x 70um; mobile phase: [H2O (10mM NH4HCO3)-ACN]; gradient: 5 35%-65% B over 8.0 min) to give BXV-41 (2.5 mg, 1.87 μmol, 5.43% yield, 100% purity) was obtained as a yellow solid.1H NMR (400 MHz, DMSO-d₆) δ12.85 (s, 1H), 11.73-11.72 (m, 1H), 9.76 (s, 1H), 8.95 (s, 1H), 8.28-8.25 (m, 1H), 8.04-8.02 (m, 1H), 7.92-7.86 (m, 1H), 7.80- 7.78 (m, 1H), 7.63-7.61 (m, 1H), 7.55-7.53 (m, 1H), 7.48-7.43 (m, 2H), 7.39-7.33 (m, 2H), 7.29 (s, 1H), 7.22 (d, J = 7.6 Hz, 1H), 7.01-6.99 (m, 1H), 6.91-6.87 (m, 2H), 6.52 (s, 1H), 5.14-5.10 10 (m, 2H), 4.96-4.90 (m, 2H), 4.52-4.44 (m, 2H), 4.41 (t, J = 5.6 Hz, 1H), 4.28 (d, J = 2.8 Hz, 1H), 3.94 (d, J = 3.2 Hz, 2H), 3.81 (s, 2H), 3.61 (s, 2H), 3.42-3.37 (m, 2H), 3.05–3.01 (m, 2H), 2.45 (s, 3H), 2.10 (s, 3H), 2.03-1.98 (m, 2H), 1.90-1.79 (m, 3H), 1.38-0.99 (m, 15H), 0.94 (s, 9H), 0.84 (s, 6H). LCMS: MS (ESI) m/z 1338.7[M+H]⁺ The following compounds were prepared in a manner similar to that described for BXV- 15 41 using the appropriate compound as starting material. 263 15077.006WO1 Example WV-6 Synthesis of (2S,4R)-1-[(2S)-2-[[2-[6-[4-[4-[[2-allyl-1-[6-(1- hydroxy-1-methyl-ethyl)-2-pyridyl]-3-oxo-pyrazolo[3,4-d]pyrimidin-6- yl]amino]phenyl]piperazin-1-yl]hexoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-N-[(1S)-1-[5- 5 fluoro-2-hydroxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]-4-hydroxy-pyrrolidine-2-carboxamide WV-6 264 15077.006WO1 Step A. Preparation of Int 9a To a solution of 6-benzyloxyhexan-1-ol (11.5 g, 55.2 mmol, 1 eq), TBACl (15.3 g, 55.2 mmol, 15.4 mL, 1 eq), and tert-butyl 2-bromoacetate (16.2 g, 82.8 mmol, 12.23 mL, 1.5 eq) in 5 toluene (207 mL) was added a solution of NaOH (57.4 g, 1.44 mol, 26 eq) in H₂O (180 mL) at 0°C, and then stirred for 12 hrs at 25°C. The reaction mixture was poured into ice-water (w/w = 1/1) (100 mL) and stirred for 5 min. The aqueous phase was extracted with ethyl acetate (100 mL x 2). The combined organic phase was washed with brine (100 mL), dried with anhydrous Na2SO4, filtered and concentrated in vacuum. The residue was purified by flash silica gel 10 chromatography (ISCO®; 220 g SepaFlash® Silica Flash Column, Eluent of 5~10% ethyl acetate/petroleum ether gradient @ 200 mL/min). Compound tert-butyl 2-(6- benzyloxyhexoxy)acetate Int 9a (8.4 g, 26 mmol, 47.19% yield) was obtained as colorless oil. 1H NMR: (400 MHz, CDCl3) δ7.28 - 7.24 (m, 4H), 7.23 - 7.17 (m, 1H), 4.43 (s, 2H), 4.09 - 3.87 (s, 2H), 3.48 - 3.34 (m, 4H), 1.61 - 1.48 (m, 4H), 1.42 - 1.40 (m, 9H), 1.35 - 1.26 (m, 4H). 15 Step B. Preparation of Int 9b A mixture of Int 9a (8.4 g, 26.1 mmol, 1 eq), Pd/C (4.20 g, 3.95 mmol, 10% purity) in EtOH (200 mL) was degassed and purged with H2 for 3 times, and then stirred at 25 °C for 12hr under H₂ atmosphere(50Psi). The mixture was filtered by cilte, then the filtrate was concentrated in vacuum. The residue was purified by flash silica gel chromatography (ISCO®; 20 80 g SepaFlash® Silica Flash Column, Eluent of 35~40% Ethyl acetate/Petroleum ethergradient @ 150 mL/min). Compound tert-butyl 2-(6-hydroxyhexoxy)acetate Int 9b (4.27 g, 18.4 mmol, 265 15077.006WO1 70.55% yield) was obtained as colorless oil.¹ H NMR (400 MHz, CDCl3) δ3.93 (s, 2H), 3.62 (t, J = 6.4 Hz, 2H), 3.50 (t, J = 6.8 Hz, 2H), 1.66 - 1.51 (m, 4H), 1.46 (s, 9H), 1.42 - 1.33 (m, 4H). Step C. Preparation of Int 9c To a solution of Int 9b (4.2 g, 18.1 mmol, 1 eq) in DCM (40 mL) was added Dess- 5 Martin (10.0 g, 23.4 mmol, 1.3 eq) at 25°C, and then stirred at 25 °C for 1hr. The reaction mixture was poured into sat.NaHCO₃ (100 mL). The aqueous phase was extracted with DCM (40 mL x 3). The combined organic phase was dried with anhydrous Na2SO4, filtered and concentrated in vacuum. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® Silica Flash Column, Eluent of 10~20% Ethyl acetate/Petroleum ethergradient 10 @ 120 mL/min). Compound tert-butyl 2-(6-oxohexoxy)acetate Int 9c (3.98 g, 17.2 mmol, 95.59% yield) was obtained as colorless oil.¹H NMR: (400 MHz, CDCl3) δ9.77 (s, 1H), 3.94 (s, 2H), 3.51 (t, J = 6.4 Hz, 2H), 2.47 - 2.39 (m, 2H), 1.71 - 1.60 (m, 4H), 1.48 (s, 9H), 1.45 - 1.39 (m, 2H) Step D. Preparation of Int 9d 15 To a solution of Int 9c (1.06 g, 4.62 mmol, 1.5 eq) in DCM (20 mL) were added TEA (935 mg, 9.25 mmol, 1.3 mL, 3 eq), then the pH of the mixture was adjusted to 5-6 by AcOH, after that, NaBH(OAc)3 (980 mg, 4.62 mmol, 1.5 eq) was added, and stirred at 25°C for 1hr. The resulting mixture was quenched with sat.NaHCO3 (50 mL), the aqueous phase was extracted with DCM (20 mL x 3). The combined organic phase was washed with brine (20 mL), dried 20 with anhydrous Na₂SO₄, filtered and concentrated in vacuum. The residue was purified by flash silica gel chromatography (ISCO®; 80g SepaFlash® Silica Flash Column, Eluent of 90~100% Ethyl acetate/Petroleum ethergradient @ 80 mL/min). Compound tert-butyl 2-[6-[4-[4-[[2-allyl- 1-[6-(1-hydroxy-1-methyl-ethyl)-2-pyridyl]-3-oxo-pyrazolo[3,4-d]pyrimidin-6- yl]amino]phenyl]piperazin-1-yl]hexoxy]acetate Int 9d (3.8 g, crude) was obtained as yellow oil. 25 ¹H NMR: (400 MHz, DMSO-d6) δ8.82 (s, 1H), 8.07-8.04 (m, 1H), 7.82 - 7.69 (m, 1H), 7.65 - 7.47 (m, 3H), 6.91 (d, J = 8.8 Hz, 2H), 5.76 - 5.52 (m, 1H), 4.99 (d, J = 10.0 Hz, 1H), 4.82 (d, J = 17.2 Hz, 1H), 4.68 (d, J = 4.8 Hz, 2H), 3.93 (s, 2H), 3.42 (t, J = 6.4 Hz, 2H), 3.11-3.08 (m, 4H), 2.33-2.28 (m, 2H), 1.48-1.43 (m, 6H), 1.44-1.40 (m, 13H), 1.33-1.28 (m, 4H). LCMS: MS (ESI) m/z 701.4 [M+H]⁺ 30 Step E. Preparation of Int 9e To a solution of Int 9d (3.8 g, 5.42 mmol, 1 eq) in DCM (40 mL) was added TFA (25.3 g, 222 mmol, 16.5 mL, 41 eq) at 25°C, and then stirred at 25 °C for 1hr. The reaction mixture was concentrated in vacuum. The residue was purified by prep-HPLC (column: Welch Xtimate C18250x100mm#10um;mobile phase: [H2O(0.1%TFA)-ACN];gradient:15%-50% B over 20.035 min). Compound 2-[6-[4-[4-[[2-allyl-1-[6-(1-hydroxy-1-methyl-ethyl)-2-pyridyl]-3-oxo- 266 15077.006WO1 pyrazolo[3,4-d]pyrimidin-6-yl]amino]phenyl]piperazin-1-yl]hexoxy]acetic acid Int 9e (3.3 g, 4.13 mmol, 76.20% yield, 95% purity, TFA) was obtained as a yellow solid.¹H NMR: (400 MHz, DMSO-d₆) δ10.20 (s, 1H), 9.51 (s, 1H), 8.85 (s, 1H), 8.04 (t, J = 7.6 Hz, 1H), 7.75 (d, J = 8.0 Hz, 1H), 7.61 (d, J = 7.6 Hz, 3H), 7.01 (d, J = 9.2 Hz, 2H), 5.72 - 5.60 (m, 1H), 5.02 - 4.97 5 (m, 1H), 4.85 - 4.79 (m, 1H), 4.68 (d, J = 4.8 Hz, 2H), 3.98 (s, 2H), 3.79 (d, J = 12.8 Hz, 2H), 3.60 (d, J = 11.6 Hz, 2H), 3.45 (t, J = 6.4 Hz, 2H), 3.15 (d, J = 11.6 Hz, 4H), 3.00 - 2.90 (m, 2H), 1.74 - 1.64 (m, 2H), 1.57 - 1.51 (m, 2H), 1.46 (s, 6H), 1.35 (d, J = 2.4 Hz, 4H). LCMS: MS (ESI) m/z 645.4 [M+H]⁺ Step F. Preparation of WV-6 10 To a solution of Int 9e (70 mg, 108 μmol, 1 eq) in DMF (2 mL) were added (2S,4R)-1- [(2S)-2-amino-3,3-dimethyl-butanoyl]-N-[(1S)-1-[5-fluoro-2-hydroxy-4-(4-methylthiazol-5- yl)phenyl]ethyl]-4-hydroxy-pyrrolidine-2-carboxamide (67.1 mg, 130 μmol, 1.2 eq, HCl), DIEA (70.2 mg, 542 μmol, 94.5 μL, 5 eq) and HATU (61.9 mg, 162 μmol, 1.5 eq) at 25°C, and then 15 stirred for 1hr. The reaction mixture was quenched by addition TFA (0.2 mL) until pH=5-6, and purified by prep-HPLC (column: Phenomenex Luna C1875x30mmx3um;mobile phase: [H₂O(0.1% TFA)-ACN];gradient:20%-50% B over 8.0 min) to give WV-6 (18.3 mg, 16.3 μmol, 14.9% yield, 98.22% purity) as a yellow solid.1H NMR (400 MHz, DMSO-d₆) δ9.02 (s, 1H), 8.81 (s, 1H), 8.02 - 7.98 (m, 1H), 7.70 (d, J = 8.0 Hz, 1H), 7.58 (d, J = 7.2 Hz, 2H), 7.33 (d, J = 267 15077.006WO1 9.6 Hz, 1H), 7.03 (d, J = 10.8 Hz, 1H), 6.98 (d, J = 9.2 Hz, 2H), 6.80 (d, J = 6.4 Hz, 1H), 5.68 - 5.59 (m, 1H), 5.03 (d, J = 6.8 Hz, 1H), 4.99 - 4.97 (m, 1H), 4.80 - 4.76 (m, 1H), 4.65 (d, J = 4.0 Hz, 2H), 4.54 - 4.51 (m, 1H), 4.43 - 4.37 (m, 1H), 4.29 (s, 1H), 3.90 (s, 1H), 3.85 (s, 2H), 3.80 (t, J = 4.8 Hz, 2H), 3.59 - 3.55 (m, 4H), 3.49 - 3.46 (m, 2H), 3.10 (d, J = 8.8 Hz, 4H), 2.95 - 2.89 5 (m, 2H), 2.46 (s, 1H), 2.30 (s, 3H), 2.16 - 2.09 (m, 1H), 1.79 - 1.74 (m, 1H), 1.67 (s, 2H), 1.55 (d, J = 6.4 Hz, 2H), 1.43 (s, 6H), 1.37 (d, J = 6.4 Hz, 4H), 1.29 (d, J = 7.2 Hz, 3H), 0.91 (s, 9H). LCMS: MS (ESI) m/z 1105.6[M+H]⁺ The following compounds were prepared in a manner similar to that described for WV-6 using the appropriate compound as starting material. 10 Example WV-18 Synthesis of (2S,4R)-1-[(2S)-2-[4-[6-[4-[4-[[2-allyl-1-[6-(1- hydroxy-1-methyl-ethyl)-2-pyridyl]-3-oxo-pyrazolo[3,4-d]pyrimidin-6- yl]amino]phenyl]piperazin-1-yl]hexoxy]triazol-1-yl]-3-methyl-butanoyl]-4-hydroxy-N-[(1S)-1- [2-hydroxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide, WV-18 15 Step A. Preparation of Int 10a 268 15077.006WO1 To a solution of 6-[tert-butyl(dimethyl)silyl]oxyhexan-1-ol (2 g, 8.60 mmol, 1 eq) in THF (20 mL) was added NaH (516 mg, 12.9 mmol, 60% purity, 1.5 eq) at 0°C in portions. The mixture was stirred at 0°C for 0.5 hr. Then 1,1,2-trichloroethylene (2.26 g, 17.2 5 mmol, 2.26 mL, 2 eq) was added dropwise at 0°C. The reaction mixture was warmed to 25°C and stirred for 2 hr. The mixture was added water (20 mL) at 0°C and stirred for 10 min, then the mixture was extracted with ethyl acetate (30 mL x 3). The combined organic phases were washed with brine (20 mL), dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to dryness. The residue was purified by column 10 chromatography (SiO2, Petroleum ether/Ethyl acetate=100/0 to 98/2) to give tert-butyl-[6-[(Z)- 1,2-dichlorovinyloxy]hexoxy]-dimethyl-silane Int 10a (1.5 g, 4.58 mmol, 53.2% yield) as colourless oil.1H NMR (400 MHz, DMSO-d6) δ6.08 (s, 1H), 4.01 (t, J = 6.4 Hz, 2H), 3.59 - 3.55 (m, 2H), 1.65 - 1.56 (m, 2H), 1.47 - 1.28 (m, 6H), 0.86 (s, 9H), 0.02 (s, 6H) Step B. Preparation of Int 10b 15 To a solution of Int 10a (1.5 g, 4.58 mmol, 1 eq) in THF (10 mL) was added dropwise n- BuLi (2.5 M, 4.58 mL, 2.5 eq) at -78°C . The reaction mixture was warmed to -50°C and stirred for 1 hr. After warmed to 0°C, the reaction mixture was quenched by addition of NH4Cl (20 mL) and H₂O 20 mL, then extracted with EtOAc (30 mL x 3). The combined organic layers 20 were washed with brine 20 mL, dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column to give tert-butyl-(6- ethynoxyhexoxy)-dimethyl-silane Int 10b (1.4 g, 4.37 mmol, 95.3% yield, 80% purity) as colourless oil.1H NMR (400 MHz, DMSO-d6) δ4.08-3.95(m, 2H), 3.57 (t, J = 6.4 Hz, 2H), 1.69 - 1.45 (m, 4H), 1.40 (s, 1H), 1.32 -1.30 (m, 4H), 0.86 (s, 9H), 0.02 (s, 6H) 25 Step C. Preparation of Int 10c 269 15077.006WO1 To a solution of Int 10b (1 g, 2.12 mmol, 1 eq) in THF:t-BuOH:H2O=1:1:1 (9 mL) were added tert-butyl-(6-ethynoxyhexoxy)-dimethyl-silane (814 mg, 3.17 mmol, 1.5 eq), CuSO4 (168 mg, 1.06 mmol, 162 μL, 0.5 eq) and sodium;(2R)-2-[(1S)-1,2-dihydroxyethyl]-4-hydroxy-5- 5 oxo-2H-furan-3-olate (628 mg, 3.17 mmol, 1.5 eq). The reaction mixture was heated to 30°C and stirred for 2 hr. After cooling to room temperature, water (30 mL) was added and then extracted with DCM (30 mL x 3). The combined organic phases were washed with water (20 mL), brine (20 mL), dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to dryness. The residue was purified by column chromatography (SiO₂, 10 EtOAc/MeOH=100/0 to 50/50) to give (2S,4R)-1-[(2S)-2-[4-[6-[tert- butyl(dimethyl)silyl]oxyhexoxy]triazol-1-yl]-3-methyl-butanoyl]-4-hydroxy-N-[(1S)-1-[2- hydroxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide Int 10c (0.8 g, 1.10 mmol, 51.86% yield) as a brown solid. LCMS: MS (ESI) m/z 729.4[M+H]⁺ Step D. Preparation of Int 10d 15 To a solution of Int 10c (0.3 g, 411 μmol, 1 eq) in DCM (3 mL) were added Ac2O (105 mg, 1.03 mmol, 96.6 μL, 2.5 eq), DMAP (5.0 mg, 41.1 μmol, 0.1 eq) and TEA (166 mg, 1.65 mmol, 229 μL, 4 eq). The reaction mixture was stirred at 25°C for 1 hr. The mixture was added water (10 mL) and extracted with DCM (30 mL x 3), dried over anhydrous Na2SO4,20 filtered and concentrated under reduced pressure to give [2-[(1S)-1-[[(2S,4R)-4-acetoxy-1-[(2S)- 2-[4-[6-[tert-butyl(dimethyl)silyl]oxyhexoxy]triazol-1-yl]-3-methyl-butanoyl]pyrrolidine-2- 270 15077.006WO1 carbonyl]amino]ethyl]-5-(4-methylthiazol-5-yl)phenyl] acetate Int 10d (0.32 g, crude) as brown oil. LCMS: MS (ESI) m/z 813.5[M+H]⁺ Step E. Preparation of Int 10e 5 To a solution of Int 10d (0.32 g, 393 μmol, 1 eq) in MeOH (3 mL) was added dropwise acetyl chloride (46.3 mg, 590 μmol, 41.9 μL, 1.5 eq). The reaction mixture was stirred at 25°C for 1 hr. The reaction mixture was added H2O (10 mL x 3) and extracted with ethyl acetate (20 mL x 3), the organic layer was washed with brine (10 mL), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give [2-[(1S)-1-[[(2S,4R)-4-acetoxy-1-[(2S)-10 2-[4-(6-hydroxyhexoxy)triazol-1-yl]-3-methyl-butanoyl]pyrrolidine-2-carbonyl]amino]ethyl]-5- (4-methylthiazol-5-yl)phenyl] acetate Int 10e (0.29 g, crude) as brown oil. LCMS: MS (ESI) m/z 699.3[M+H]⁺ Step F. Preparation of Int 10f 15 To a solution of Int 10e (0.26 g, 372 μmol, 1 eq) in DCM (3 mL) was added Dess-Martin (473 mg, 1.12 mmol, 345 μL, 3 eq) .The reaction mixture was stirred at 25°C for 1 hr. The mixture was added water (10 mL) and extracted with DCM (30 mL x 3), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to dryness. The residue was purified by prep-TLC (SiO₂, Petroleum ether/Ethyl acetat = 0:1) to give [2-[(1S)-1-[[(2S,4R)-4-acetoxy-20 1-[(2S)-3-methyl-2-[4-(6-oxohexoxy)triazol-1-yl]butanoyl]pyrrolidine-2-carbonyl]amino]ethyl]- 5-(4-methylthiazol-5-yl)phenyl] acetate Int 10f (0.09 g, 129 μmol, 34.7% yield) as a brown solid. LCMS: MS (ESI) m/z 697.4[M+H]⁺ Step G. Preparation of Int 10g 271 15077.006WO1 To a solution of 2-allyl-1-[6-(1-hydroxy-1-methyl-ethyl)-2-pyridyl]-6-(4-piperazin-1- ylanilino)pyrazolo[3,4-d]pyrimidin-3-one (0.035 g, 66.9 μmol, 1 eq, HCl) in DCM (1 mL) was added TEA (20.3 mg, 200 μmol, 27.9 μL, 3 eq), then AcOH (8.0 mg, 133 μmol, 7.66 μL, 5 2 eq) was added to adjust pH to 5~6, then Int 10f (51.2 mg, 73.6 μmol, 1.1 eq) and NaBH(OAc)₃ (21.2 mg, 100 μmol, 1.5 eq) were added to the mixture. The reaction mixture was stirred at 25°C for 1 hr. The reaction mixture was added H₂O 10 mL and extracted with ethyl acetate (20 mL x 3), the organic layer was washed with brine (10 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give [2-[(1S)-1-[[(2S,4R)-4-10 acetoxy-1-[(2S)-2-[4-[6-[4-[4-[[2-allyl-1-[6-(1-hydroxy-1-methyl-ethyl)-2-pyridyl]-3-oxo- pyrazolo[3,4-d]pyrimidin-6-yl]amino]phenyl]piperazin-1-yl]hexoxy]triazol-1-yl]-3-methyl- butanoyl]pyrrolidine-2-carbonyl]amino]ethyl]-5-(4-methylthiazol-5-yl)phenyl] acetate Int 10g (0.09 g, crude) as yellow gum. LCMS: MS (ESI) m/z 584.6[1/2M+H]⁺ Step H. Preparation of Compound WV-18 272 15077.006WO1 To a solution of Int 10g (0.085 g, 72.8 μmol, 1 eq) in MeOH (2 mL) was added K₂CO₃ (30.1 mg, 218 μmol, 3 eq). The reaction mixture was stirred at 25°C for 1 hr. The mixture was acidified with TFA (PH=5~6) and filtered, then the filtrate was purified by pre-HPLC (TFA 5 condition; column: Phenomenex Luna C1875*30mm*3um;mobile phase: [H₂O(0.1% TFA)- ACN];gradient:15%-45% B over 8.0 min) to give WV-18 (32.4 mg, 29.91 μmol, 41.08% yield) as a yellow solid.1H NMR (400 MHz, DMSO-d6) δ8.93 (s, 1H), 8.83 (s, 1H), 7.99-8.06 (m, 1H), 7.73 (d, J = 7.6 Hz, 1H), 7.68 (s, 1H), 7.58-7.64 (m, 3H), 7.18 (d, J = 8.0 Hz, 1H), 6.99 (d, J = 9.2 Hz, 2H), 6.85-6.94 (m, 2H), 5.58-5.71 (m, 1H), 5.07-5.13 (m, 2H), 4.99 (d, J = 10.8 10 Hz, 1H), 4.81 (d, J = 17.2 Hz, 1H), 4.67 (d, J = 4.8Hz, 2H), 4.39 (t, J = 8.4 Hz, 1H), 4.31 (d, J = 1.6 Hz, 1H), 4.04-4.12 (m, 2H), 3.70-3.82 (m, 4H), 3.09-3.22 (m, 5H), 2.93 (t, J = 12.0 Hz, 2H), 2.44 (s, 3H), 2.04-2.12 (m, 1H), 1.65-1.82 (m, 6H), 1.45 (s, 9H), 1.35-1.40 (m, 2H), 1.30 (d, J = 7.2 Hz, 3H),1.03 (d, J = 6.4 Hz, 3H), 0.67 (d, J = 6.4 Hz, 3H). LCMS: MS (ESI) m/z 1083.6[M+H]⁺ 15 The following compounds were prepared in a manner similar to that described for WV- 18 using the appropriate compound as starting material. 273 15077.006WO1 Example WV-13 Synthesis of (2S,4R)-1-((S)-2-(2-((6-(4-(4-((2-allyl-1-(6-(2- hydroxypropan-2-yl)pyridin-2-yl)-3-oxo-2,3-dihydro-1H-pyrazolo[3,4-d]pyrimidin-6- yl)amino)phenyl)piperazin-1-yl)hexyl)oxy)acetamido)-3,3-dimethylbutanoyl)-N-((8-chloro-1- 5 oxo-1,2-dihydrophthalazin-5-yl)methyl)-4-hydroxypyrrolidine-2-carboxamide, WV-13 274 15077.006WO1 Step A. Preparation of Int 11a To a solution of 8-chloro-5-iodo-2H-phthalazin-1-one (5 g, 16.3 mmol, 1 eq) in dioxane (50 mL) and H₂O (10 mL) was added potassium;(tert-butoxycarbonylamino)methyl-trifluoro- boranide (5.80 g, 24.4 mmol, 1.5 eq), Cs2CO3 (10.6 g, 32.6 mmol, 2 eq) and [2-(2- 5 aminophenyl)phenyl]-chloro-palladium;bis(1-adamantyl)-butyl-phosphane (1.09 g, 1.63 mmol, 0.1 eq) at 25°C. The reaction mixture was degassed and purged with N₂ for 3 times and heated to 110°C and stirred for 16 hrs. After cooling to room temperature, the reaction mixture was quenched by addition H2O (200 mL) and then extracted with EtOAc(50 mL x 3). The combined organic layers were washed with brine (50 mL x 2), dried over Na₂SO₄, filtered and concentrated 10 under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, Eluent of 60~80% Ethyl acetate/Petroleum ethergradient @ 150 mL/min). Compound tert-butyl N-[(8-chloro-1-oxo-2H- phthalazin-5-yl)methyl]carbamate Int 11a (4.9 g, crude) was obtained as a white solid. LCMS: MS (ESI) m/z 310.0 [M+H]⁺ 15 Step B. Preparation of Int 11b To a solution of Int 11a (2.9 g, 9.36 mmol, 1 eq) in EtOAc (10 mL) was added HCl/EtOAc (4 M, 30 mL, 12.8 eq) at 25°C. The reaction mixture was stirred at 25°C for 1 hr. The reaction mixture was filtered and the filter cake was concentrated under reduced pressure to give a residue. Compound 5-(aminomethyl)-8-chloro-2H-phthalazin-1-one Int 11b (2 g, crude) 20 was obtained as a white solid. LCMS: MS (ESI) m/z 461.1[M+H]⁺ Step C. Preparation of Int 11c To a solution of Int 11b (1 g, 4.77 mmol, 1 eq) in DMF (10 mL) was added (2S,4R)-1- [(2S)-2-(tert-butoxycarbonylamino)-3,3-dimethyl-butanoyl]-4-hydroxy-pyrrolidine-2-carboxylic acid (2.46 g, 7.16 mmol, 1.5 eq), DIEA (3.08 g, 23.8 mmol, 4.15 mL, 5 eq) and HATU (2.72 g, 25 7.16 mmol, 1.5 eq) at 25°C. The reaction mixture was stirred at 25°C for 1 hr. The reaction mixture was quenched by addition H₂O (30 mL), and then extracted with EtOAc (50 mL x 3). The combined organic layers were washed with brine (50 mL x 2), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The crude product was purified by reversed-phase HPLC (MeCN/H₂O condition, 130ml/min, 35-65% 25min; 65% 10min). 30 Compound tert-butyl N-[(1S)-1-[(2S,4R)-2-[(8-chloro-1-oxo-2H-phthalazin-5- yl)methylcarbamoyl]-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]carbamate Int 11c (2.8 g, crude) was obtained as a white solid.¹H NMR (400 MHz, DMSO-d6) δ12.69 (s, 1H), 8..72 (t, J = 5.6 Hz, 1H), 8.44 (s, 1H), 7.89 (d, J = 8.0 Hz, 1H), 7.69 (d, J = 8.0 Hz, 1H), 6.46 (d, J = 9.2 Hz, 1H), 5.15 (s, 1H), 4.70-4.65 (m, 1H), 4.60-4.55 (1, 2H), 4.48-4.43 (m, 1H), 4.36- 275 15077.006WO1 4.34 (m, 1H), 4.15-4.10 (m, 1H), 3.65-3.60 (m, 1H), 2.10-2.06 (m, 1H), 1.90-1.80 (m, 1H), 1.37(s, 9H), 0.88 (s, 9H). LCMS: MS (ESI) m/z 536.1 [M+H]⁺ Step D. Preparation of Int 11d To a solution of Int 11c (1.8 g, 3.36 mmol, 1 eq) in EtOAc (2 mL) was added 5 HCl/EtOAc (4 M, 20 mL, 23.8 eq) in a portion at 25°C. The reaction mixture was stirred at 25°C for 1 hr. The reaction mixture was filtered and the filter cake was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: Phenomenex luna C18250 x 50mm x 15um;mobile phase: [H2O(0.1% TFA)- ACN];gradient:5%-25% B over 10.0 min). Compound (2S,4R)-1-[(2S)-2-amino-3,3-dimethyl-10 butanoyl]-N-[(8-chloro-1-oxo-2H-phthalazin-5-yl)methyl]-4-hydroxy-pyrrolidine-2- carboxamide Int 11d (1.34 g, 3.07 mmol, 91.54% yield) was obtained as a white solid.¹H NMR (400 MHz, DMSO-d₆) δ12.70 (s, 1H), 8..92 (d, J = 4.2 Hz, 1H), 8.45 (s, 1H), 8.19 (s, 2H), 7.75 (d, J = 8.4 Hz, 1H), 7.69 (d, J = 8.4 Hz, 1H), 4.70-4.65 (m, 1H), 4.56-4.51 (m, 2H), 4.35 (s, 1H), 3.87-3.85 (m, 1H), 3.77 (d, J = 5.2 Hz, 2H), 3.56-3.52 (m, 1H), 2.50-2.06 (m, 1H), 1.87-1.58 (m, 15 1H), .98 (s, 9H). Step E. Preparation of Int 11e To a solution of Int 9e (500 mg, 775 μmol, 1 eq) in DMF (5 mL) was added 2,3,5,6- tetrafluorophenol (386 mg, 2.33 mmol, 3 eq), and EDCI (743 mg, 3.88 mmol, 5 eq) at 25°C. 20 The reaction mixture was stirred at 25°C for 1 hr. The reaction mixture was quenched by addition H2O (30mL) and then extracted with EtOAc (10 mL x 3). The combined organic layers were washed with brine (10 mL x 2), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. Compound (2,3,5,6-tetrafluorophenyl) 2-[6-[4-[4-[[2-allyl-1- [6-(1-hydroxy-1-methyl-ethyl)-2-pyridyl]-3-oxo-pyrazolo[3,4-d]pyrimidin-6- 25 yl]amino]phenyl]piperazin-1-yl]hexoxy]acetate Int 11e (600 mg, crude) was obtained as yellow oil. LCMS: MS (ESI) m/z 793.4[M+H]⁺ Step F. Preparation of WV-13 276 15077.006WO1 To a solution of Int 11e (60 mg, 75.7 μmol, 1 eq) in DMF (1 mL) was added (2S,4R)-1- [(2S)-2-amino-3,3-dimethyl-butanoyl]-N-[(8-chloro-1-oxo-2H-phthalazin-5-yl)methyl]-4- hydroxy-pyrrolidine-2-carboxamide (39.6 mg, 90.8 μmol, 1.2 eq), DIEA (48.9 mg, 378 μmol, 5 65.9 μL, 5 eq) at 25°C. The reaction mixture was stirred at 25°C for 1 hr. The reaction mixture was quenched by addition TFA (0.2 mL) until pH to 5-6. The residue was purified by prep- HPLC (column: Phenomenex Luna C1875 x 30mm x 3um; mobile phase: [H₂O(0.1% TFA)- ACN];gradient:15%-45% B over 8.0 min) to give WV-13 (25.2 mg, 23 μmol, 30.41% yield) as a yellow solid.¹H NMR (400 MHz, DMSO-d6) δ 8.82 (s, 1H), 8.71 (t, J = 5.2 Hz, 1H), 8.41 (s, 10 1H), 8.03-8.01 (m, 1H), 7.82 (d, J = 8.4 Hz, 1H), 7.72-7.67 (m, 2H), 7.60-7.58 (m, 3H), 7.33 (d, J = 9.2 Hz, 1H), 6.99-6.97 (m, 2H), 5.69-5.60 (m, 1H), 4.99 (d, J = 10.4 Hz, 1H), 4.80 (d, J = 17.2 Hz, 1H), 4.66 (d, J = 5.6 Hz, 2H), 4.63-4.58 (m, 1H), 4.52 (d, J = 9.2 Hz, 1H), 4.40-4.37 (m, 1H), 4.34 (s, 1H), 4.01 (s, 1H), 3.91 (s, 2H), 3.77 (d, J = 12.4 Hz, 2H), 3.63 (s, 1H), 3.61- 3.55 (m, 4H), 3.49-3.45 (m, 3H), 3.15 (s, 1H), 3.11 (d, J = 8.4 Hz, 3H), 2.95-2.92 (m, 2H), 2.07 -15 1.98 (m, 1H), 1.90-1.82 (m, 1H), 1.66 (d, J = 5.6 Hz, 2H), 1.58-1.55 (m, 2H), 1.44 (s, 6H), 1.36- 1.34 (m, 4H), 0.89-0.88 (m, 9H). LCMS: MS (ESI) m/z 1062.5[M+H]⁺ Example 200 Synthesis of Linker-payloads Step A. Preparation of Int 50a 277 15077.006WO1 To a solution of tert-butyl methylglycinate (100 g, 550 mmol, 1.00 eq, hydrochloride) and N-(((9H-fluoren-9-yl)methoxy)carbonyl)-N-methylglycine (171 g, 550 mmol, 1.00 eq) in dimethyl formamide (400 mL) was added diisopropylethylamine (142 g, 1.10 mol, 192 mL, 2.00 5 eq) and O-(7-azabenzotriazol-1-yl)-N,N,N’,N’-tetramethyluronium hexafluorophosphate (251 g, 661 mmol, 1.20 eq). Then the mixture was stirred at 25 °C for 1 h. The reaction mixture was poured into water (3 L) and extracted with ethyl acetate (3 × 200 mL). The organic layer was washed with brine (3 × 200 mL) and concentrated to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 330 g SepaFlash ® Silica Flash Column, Eluent of10 30~90% ethyl acetate/petroleum ether gradient @ 120 mL/min) to afford tert-butyl N-(N-(((9H- fluoren-9-yl)methoxy)carbonyl)-N-methylglycyl)-N-methylglycinate, Int 50a, (230 g, 524 mmol, 95% yield) as yellow oil.¹H NMR (400 MHz, DMSO-d₆) δ = 7.89 (t, J = 8.4 Hz, 2H), 7.69 - 7.56 (m, 2H), 7.41 (q, J = 7.4 Hz, 2H), 7.37 - 7.25 (m, 2H), 4.33 - 4.08 (m, 4H), 4.04 - 3.93 (m, 3H), 2.97 - 2.79 (m, 6H), 1.45 - 1.35 (m, 9H). 15 Step B. Preparation of Int 50b A solution of Int 50a, (230 g, 524 mmol, 1.00 eq) in trifluoroacetic acid (500 mL) and dichloromethane (500 mL) was stirred at 25 °C for 3 h. The reaction mixture was concentrated to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 330 g 20 SepaFlash® Silica Flash Column, Eluent of 50~100% ethyl acetate/petroleum ether gradient @ 120 mL/min) to afford N-(N-(((9H-fluoren-9-yl)methoxy)carbonyl)-N-methylglycyl)-N- methylglycine, Int 50b, (200 g, 523 mmol, 99% yield) as yellow oil.¹H NMR (400 MHz, DMSO-d₆) δ = 7.89 (br t, J = 8.8 Hz, 2H), 7.72 - 7.56 (m, 2H), 7.41 (q, J = 7.2 Hz, 2H), 7.37 - 7.25 (m, 2H), 4.35 - 4.23 (m, 2H), 4.23 - 4.08 (m, 2H), 4.07 - 3.97 (m, 3H), 3.01 - 2.77 (m, 6H). 25 Step C. Preparation of Int 50d 278 15077.006WO1 To a solution of Int 50b, (35.0 g, 91.5 mmol, 1.00 eq) in dichloromethane (600 mL) was added to 2-chlorotrityl chloride resin (175 g). Then diisopropylethylamine (29.6 g, 228 mmol, 39.9 mL, 2.50 eq) was added. The mixture was stirred at 25°C for 2 h. The mixture was filtered 5 and washed with dimethyl formamide (600 mL) and dichloromethane/methanol/diisopropylethylamine (600 mL, 80:15:5). The mixture was filtered and washed with dimethyl formamide (3 × 600 mL) and dichloromethane (3 × 600 mL). This reaction was in parallel twice. The resin, Int 50c, was stored at 0 °C for next step. Elongation of the polysarcosine oligomer was performed until the desired length was 10 obtained, by alternating remove Fmoc and amide coupling steps. In the first step, the resin, Int 50c, was treated with piperidine (50%) in dimethyl formamide at 25°C for 1 min and washed with dimethyl formamide (4 times). For the amide coupling step, to the resin was added a solution of N-(N-(((9H-fluoren-9-yl)methoxy)carbonyl)-N-methylglycyl)-N-methylglycine (1.30 eq), O-(7-azabenzotriazol-1-yl)-N,N,N’,N’-tetramethyluronium hexafluorophosphate (1.50 eq) 15 and diisopropylethylamine (2 eq) in dimethylformamide. The reaction vessel was stirred at 25 °C for 2 h and the resin was extensively washed with dimethyl formamide (4 times) and dichloromethane (3 times). The resin, Int 50d, was dried under vacuum and stored at 0 °C. Step D. Preparation of Int 50e 279 15077.006WO1 The resin, Int 50d, in piperidine (500 mL) and dimethyl formamide (500 mL) was agitated with nitrogen atmosphere at 15 °C for 1 min. The mixture was filtered and washed with dimethyl formamide (3 × 500 mL) and dichloromethane (500 mL), then the filter cake was 5 concentrated in vacuum. The resin, Int 50e, (the equivalent of 50 g of Int 50f loaded on resin) was dried under vacuum and stored at 0 °C. Step E. Preparation of Int 50f The resin, Int 50d, (42 g) in piperidine (40.0 mL) and dimethyl formamide (160 mL) 10 was agitated with nitrogen atmosphere for 1 h at 25°C. Then the solution was drained and the resin was washed with dimethyl formamide (3 × 120 mL) and dichloromethane (3 × 120 mL). Then it was agitated in 1,1,1,3,3,3-Hexafluro-2-propanol (40 mL) and DCM (160 mL) with nitrogen atmosphere for 1 h at 25 °C and filtered. The filtrate was concentrated to give a residue, which was purified by prep-HPLC (column: Waters Atlantis T3 15 150*30mm*5um;mobile phase: [water(FA)-ACN];gradient:1%-20% B over 10 min) to give 280 15077.006WO1 5,8,11,14,17,20,23,26,29-nonamethyl-4,7,10,13,16,19,22,25,28-nonaoxo- 2,5,8,11,14,17,20,23,26,29-decaazahentriacontan-31-oic acid, Int 50f, (550 mg, 754 μmol) as a white solid.¹H NMR (400 MHz, DMSO-d₆) δ = 4.50 - 3.50 (m, 24H), 2.95 - 270 (m, 23H), 2.49 - 2.37 (m, 3H). 5 Step A. Preparation of Int 36a To a solution of 2-fluoro-3-methoxyaniline (11.0 g, 77.9 mmol, 1.00 equiv) in N,N- 10 dimethyl formamide (40.0 mL) was added a solution of 1-bromopyrrolidine-2, 5-dione (13.9 g, 78.0 mmol, 1.00 equiv) dropwise. The mixture was stirred at 15 °C for 3 h. The mixture was diluted with brine (300 mL) and extracted with ethyl acetate (3 × 300 mL). The organic layers were collected and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give a crude product, which was purified by column 15 chromatography on silica gel (SiO2, petroleum ether/ethyl acetate=1/0 to 3/1) and concentrated to afford 4-bromo-2-fluoro-3-methoxyaniline, Int.36a, (26.0 g, 118 mmol, 75% yield) as yellow oil.¹H NMR (400 MHz, DMSO-d6) δ = 7.03 (dd, J = 8.8, 1.2 Hz, 1H), 6.47 (t, J = 8.8 Hz, 1H), 5.38 (s, 2H), 3.86 - 3.78 (m, 3H). Step B. Preparation of Int 36b 281 15077.006WO1 To a solution of Int.36a, (10.0 g, 45.5 mmol, 1.00 equiv) in dichloromethane (100 mL) was added borontribromide (171 g, 682 mmol, 65.7 mL, 15.0 equiv) at 0 °C. The mixture was stirred at 15 °C for 0.5 h. The reaction mixture was quenched by addition water (300 mL) at 0 °C. After filtration, the filtrate was extracted with ethyl acetate (3 × 200 mL). The organic 5 layers were collected and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to afford 3-amino-6-bromo-2-fluorophenol, Int.36b, (22.5 g, crude) as yellow oil.¹H NMR (400 MHz, DMSO-d6) δ = 9.95 - 9.35 (m, 1H), 6.90 (dd, J = 8.8, 1.6 Hz, 1H), 6.20 - 6.16 (m, 1H), 5.15 (br s, 2H). MS (ESI) m/z 205.9 [M+H]⁺ Step C. Preparation of Int 36c 10 To a solution of Int.36b, (16.0 g, 77.7 mmol, 1.00 equiv), cesium carbonate (50.6 g, 155 mmol, 2.00 equiv) and potassium iodide (1.29 g, 7.77 mmol, 0.100 equiv) in N, N-dimethyl formamide (50.0 mL) was added tert-butyl 4-bromobutanoate (19.1 g, 85.4 mmol, 1.10 equiv). The mixture was stirred at 85 °C for 12 h. The mixture was diluted with brine (300 mL) and extracted with ethyl acetate (3 × 200 mL). The organic layers were collected and dried over 15 anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give a crude product, which was purified by column chromatography on silica gel (SiO₂, petroleum ether/ethyl acetate=1/0 to 3/1) and concentrated to afford tert-butyl 4-(3-amino-6- bromo-2-fluorophenoxy)butanoate, Int.36c, (17.0 g, 48.8 mmol, 62% yield) as brown oil.¹H NMR (400 MHz, DMSO-d₆) δ = 7.03 (dd, J = 8.8, 1.6 Hz, 1H), 6.46 (t, J = 8.8 Hz, 1H), 5.36 (s, 20 2H), 3.98 - 3.93 (m, 2H), 2.43 (t, J = 7.2 Hz, 2H), 1.91 (br t, J = 6.8 Hz, 2H), 1.40 (s, 9H). Step D. Preparation of Int 36d To a solution of Int.36c, (12.0 g, 34.5 mmol, 1.00 equiv) and ethyl (E)-3-(4,4,5,5- tetramethyl-1,3,2-dioxaborolan-2-yl)acrylate (19.5 g, 86.2 mmol, 2.50 equiv) in dioxane (100 mL) and water (10.0 mL) was added 2-dicyclohexylphosphino-2,6-dimethoxybiphenyl (1.41 g, 25 3.45 mmol, 0.100 equitv), potassium phosphate (11.0 g, 51.7 mmol, 1.50 equiv) and bis(dibenzylideneacetone)-palladium(0) (1.58 g, 1.72 mmol, 0.050 equiv.). The mixture was stirred at 90 °C for 12 h under nitrogen atmosphere. The mixture was concentrated under reduced pressure to give a crude product, which was purified by column chromatography on silica gel (SiO₂, petroleum ether / ethyl acetate=1/0 to 3/1) and concentrated to afford tert-butyl 30 (E)-4-(3-amino-6-(3-ethoxy-3-oxoprop-1-en-1-yl)-2-fluorophenoxy)butanoate, Int.36d, (11.0 g, 30.0 mmol, 86% yield) as yellow solid.¹H NMR (400 MHz, DMSO-d6) δ = 7.70 (d, J = 16.0 Hz, 1H), 7.30 (dd, J = 8.8, 1.2 Hz, 1H), 6.50 (t, J = 8.4 Hz, 1H), 6.32 (d, J = 16.0 Hz, 1H), 5.87 (s, 2H), 4.14 (q, J = 7.2 Hz, 2H), 4.01 (t, J = 6.4 Hz, 2H), 2.40 (t, J = 7.2 Hz, 2H), 1.92 (quin, J = 6.8 Hz, 2H), 1.38 (s, 9H), 1.25 - 1.22 (m, 3H). 35 Step E. Preparation of Int 36e 282 15077.006WO1 A mixture of Int.36d, (5.00 g, 13.6 mmol, 1.00 equiv) in methanol (100 mL) was added palladium on activated carbon (5.00 g, 10% purity). The mixture was stirred at 25 °C for 12 h under hydrogen (50 Psi) atmosphere. The reaction mixture was filtered concentrated under reduced pressure to afford tert-butyl 4-(3-amino-6-(3-ethoxy-3-oxopropyl)-2- 5 fluorophenoxy)butanoate, Int.36e, (3.40 g, crude) as black oil.¹H NMR (400 MHz, DMSO-d6) δ = 6.64 (dd, J = 8.4, 1.6 Hz, 1H), 6.39 (t, J = 8.4 Hz, 1H), 5.01 - 4.91 (m, 2H), 4.03 (q, J = 6.8 Hz, 2H), 3.93 (t, J = 6.8 Hz, 2H), 2.74 - 2.64 (m, 2H), 2.47 - 2.44 (m, 2H), 2.39 (t, J = 6.8 Hz, 2H), 1.90 (quin, J = 6.8 Hz, 2H), 1.40 (s, 9H), 1.15 (t, J = 6.8 Hz, 3H). Step F. Preparation of Int 36f 10 To a solution of Int.36e, (5.00 g, 13.5 mmol, 1.00 equiv) in methanol (10.0 mL), tetrahydrofuran (10.0 mL) and water (10.0 mL) was added lithium hydroxide monohydrate (852 mg, 20.3 mmol, 1.50 equiv). The mixture was stirred at 15 °C for 1 h. The mixture was quenched with hydrochloric acid (1M) until pH=7. The mixture was concentrated to remove tetrahydrofuran and methanol. The mixture was diluted with water (150 mL) and extracted with 15 ethyl acetate (3 × 150 mL). The organic layers were collected and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to afford 3-(4- amino-2-(4-(tert-butoxy)-4-oxobutoxy)-3-fluorophenyl)propanoic acid, Int.36f, (4.00 g, crude) as yellow oil.¹H NMR (400 MHz, DMSO-d6) δ = 12.45 - 11.77 (m, 1H), 6.65 (dd, J = 8.4, 1.2 Hz, 1H), 6.39 (t, J = 8.4 Hz, 1H), 4.96 (s, 2H), 3.93 (t, J = 6.4 Hz, 2H), 2.66 (t, J = 7.2 Hz, 2H), 20 2.41 - 2.36 (m, 4H), 1.90 (quin, J = 6.8 Hz, 2H), 1.40 (s, 9H). Step G. Preparation of Int 36g To a solution of Int.36f, (4.00 g, 11.7 mmol, 1.00 equiv) in dichloromethane (50.0 mL) was added triethylamine (3.56 g, 35.2 mmol, 4.89 mL, 3.00 equiv) and methyl 2,5-dioxo-2,5- dihydro-1H-pyrrole-1-carboxylate (2.00 g, 12.9 mmol, 2.00 mL, 1.10 equiv) at 0 °C. The 25 mixture was stirred at 15 °C for 2 h. The mixture was diluted with water (100 mL) and extracted with dichloromethane (3 × 80 mL). The organic layers were collected and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to afford 3-(2-(4-(tert-butoxy)-4-oxobutoxy)-4-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-3- fluorophenyl)propanoic acid, Int.36g, (4.50 g, crude) as brown oil.¹H NMR (400 MHz, 30 DMSO-d₆) δ = 7.25 (s, 2H), 7.16 (d, J = 8.8 Hz, 1H), 7.08 - 7.03 (m, 1H), 4.01 (br t, J = 6.4 Hz, 2H), 2.87 (br t, J = 7.6 Hz, 2H), 2.52 - 2.51 (m, 2H), 2.42 - 2.36 (m, 4H), 1.39 (s, 9H). Step H. Preparation of Int 36h To a solution of Int.36g, (4.50 g, 10.7 mmol, 1.00 equiv) and 2,3,5,6-tetrafluorophenol (3.55 g, 21.4 mmol, 2.00 equiv) in dichloromethane (50.0 mL) and N,N-dimethylacetamide 35 (5.00 mL) was added 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (4.09 g, 283 15077.006WO1 21.4 mmol, 2.00 equiv) at 0 °C. The mixture was stirred at 15 °C for 1 h. The mixture was concentrated under reduced pressure to give a crude product, which was purified by reversed- phase HPLC (column: spherical C18, 20-45 um, 100Å, SW 330, mobile phase: [0.1% formic acid - acetonitrile]; B%: 85%-100%, 60 min). The desired fraction collected and lyophilized to 5 afford tert-butyl 4-(3-(2, 5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-fluoro-6-(3-oxo-3-(2,3,5,6- tetrafluorophenoxy)propyl)phenoxy)butanoate, Int.36h, (3.00 g, 5.27 mmol, 49 %yield) as a brown solid.¹H NMR (400 MHz, DMSO-d6) δ = 7.97 - 7.91 (m, 1H), 7.26 (s, 2H), 7.24 (s, 1H), 7.13 - 7.08 (m, 1H), 4.06 (t, J = 6.4 Hz, 2H), 3.18 - 3.12 (m, 2H), 3.10 - 3.04 (m, 2H), 2.43 - 2.39 (m, 2H), 1.97 - 1.92 (m, 2H), 1.37 (s, 9H). MS (ESI) m/z 587.1 [M+H₂O]⁺ 10 Step I. Preparation of Int 36i To a solution of Int.36h, (1.00 g, 1.76 mmol, 1.00 equiv) in dichloromethane (15.0 mL) was added trifluoroacetic acid (76.8 g, 673 mmol, 50.0 mL, 383 equiv). The mixture was stirred at 15 °C for 0.5 h. The reaction mixture was lyophilized to give a crude product, which was 15 triturated with methyl tert-butyl ether (10 mL) at 25 °C. Then filtered, the filter cake was dried in vacuum to afford 4-(3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-fluoro-6-(3-oxo-3-(2,3,5,6- tetrafluorophenoxy)propyl)phenoxy)butanoic acid, Int.36i, (300 mg, 584 μmol, 33% yield) as a pink solid.¹H NMR (400 MHz, DMSO-d₆) δ = 13.15 - 11.06 (m, 1H), 7.94 (tt, J = 7.6, 10.8 Hz, 1H), 7.26 (s, 2H), 7.24 (s, 1H), 7.15 - 7.06 (m, 1H), 4.08 (t, J = 6.0 Hz, 2H), 3.23 - 3.13 (m, 2H), 20 3.11 - 3.02 (m, 2H), 2.42 (t, J = 7.4 Hz, 2H), 1.96 (quin, J = 6.8 Hz, 2H). Step J. Preparation of Int 36j To a solution of Int.36i, (100 mg, 195 μmol, 1.00 equiv) in dichloromethane (3.00 mL) was added 1-chloro-N,N,2-trimethylprop-1-en-1-amine, Int.36j, (52.1 mg, 390 μmol, 51.5 μL, 284 15077.006WO1 2.00 eqiv). The mixture was stirred at 25 °C for 0.5 h. The reaction mixture was used in next step directly. Step K. Preparation of Int 36k 5 To a solution of 5,8,11,14,17,20,23,26,29-nonamethyl-4,7,10,13,16,19,22,25,28- nonaoxo-2,5,8,11,14,17,20,23,26,29-decaazahentriacontan-31-oic acid, Int 10f, (137 mg, 188 μmol, 1.00 equiv) in dichloromethane (2.00 mL) and N,N-diisopropylethylamine (72.9 mg, 564 μmol, 98.3 μL, 3.00 equiv). The mixture was stirred at 25 °C for 10 min. Then to the mixture was added 2,3,5,6-tetrafluorophenyl 3-(2-(4-chloro-4-oxobutoxy)-4-(2,5-dioxo-2,5-dihydro-1H- 10 pyrrol-1-yl)-3-fluorophenyl)propanoate, Int.36j, (100 mg, 188 μmol, 1.00 equiv) at 0 °C. The mixture was stirred at 25 °C for 20 min. The reaction mixture was concentrated under reduced pressure to give a crude product, which was purified by prep-HPLC (column: UniSil 3-100 C18 UItra (150*25mm*3um); mobile phase: [water (formic acid)-acetonitrile]; gradient: 20%-50% B over 40 min). The desired fraction collected and lyophilized to afford 34-(3-(2,5-dioxo-2,5-15 dihydro-1H-pyrrol-1-yl)-2-fluoro-6-(3-oxo-3-(2,3,5,6-tetrafluorophenoxy)propyl)phenoxy)- 3,6,9,12,15,18,21,24,27,30-decamethyl-4,7,10,13,16,19,22,25,28,31-decaoxo- 3,6,9,12,15,18,21,24,27,30-decaazatetratriacontanoic acid, Int.36k, (30.0 mg, 24.3 μmol, 12 % yield, 99% purity) as a white solid.¹H NMR (400 MHz, DMSO-d6) δ = 13.48 - 12.07 (m, 1H), 285 15077.006WO1 8.00 - 7.87 (m, 1H), 7.26 (s, 2H), 7.25 - 7.20 (m, 1H), 7.13 - 7.06 (m, 1H), 4.34 - 4.19 (m, 8H), 4.15 - 3.98 (m, 11H), 3.93 (br d, J = 4.4 Hz, 2H), 3.16 (br d, J = 6.0 Hz, 2H), 3.11 - 3.06 (m, 2H), 2.95 - 2.71 (m, 31H), 2.35 - 2.29 (m, 2H), 1.99 - 1.89 (m, 2H). MS (ESI) m/z 1246.4 [M+Na]⁺ 5 The following compounds were prepared in a manner similar to that described for Int 50d and Int 36k using the appropriate compound as starting material. Example RVL-1 Synthesis of 64-(6-(3-(((S)-1-(((S)-1-((4-(((((R)-2-((2S,4R)-1-((S)-10 17-((4-(benzo[d]thiazol-5-ylamino)-6-(tert-butylsulfonyl)quinolin-7-yl)oxy)-2-(tert-butyl)-4- oxo-6,9,12,15-tetraoxa-3-azaheptadecanoyl)-4-hydroxypyrrolidine-2-carboxamido)-2-(4-(4- methylthiazol-5-yl)phenyl)ethyl)(methyl)carbamoyl)oxy)methyl)phenyl)amino)-1-oxopropan-2- yl)amino)-1-oxopropan-2-yl)amino)-3-oxopropyl)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2- fluorophenoxy)-3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosamethyl-15 4,7,10,13,16,19,22,25,28,31,34,37,40,43,46,49,52,55,58,61-icosaoxo- 3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosaazatetrahexacontanoic acid, RVL-1 Step A. Preparation of Int 201a 286 15077.006WO1 To a solution of (2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[2-[[4-(1,3-benzothiazol-5-ylamino)-6- tert-butylsulfonyl-7-quinolyl]oxy]ethoxy]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethyl- butanoyl]-4-hydroxy-N-[(1R)-2-(methylamino)-1-[4-(4-methylthiazol-5- 5 yl)phenyl]ethyl]pyrrolidine-2-carboxamide, RV-2 (20 mg, 18.1 μmol, 1 eq) in DMF (0.1 mL) were added DIEA (7.03 mg, 54.3 μmol, 9.47 μL, 3 eq) and [4-[[(2S)-2-[[(2R)-2-(9H-fluoren-9- ylmethoxycarbonylamino)propanoyl]amino]propanoyl]amino]phenyl]methyl (4-nitrophenyl) carbonate (14.2 mg, 21.7 μmol, 1.2 eq). The reaction mixture was stirred at 20°C for 2 hrs. Then piperidine (7.90 mg, 92.8 μmol, 9.16 μL, 5 eq) was added. The resulting mixture was 10 stirred at 20°C for another 1 hr. The pH of mixture was adjusted to 6 with TFA, then filtered. The residue was purified by prep-HPLC (column: Phenomenex Luna C18 75*30mm*3um;mobile phase: [H₂O(0.1% TFA)-ACN];gradient:10%-40% B over 8.0 min). Afford [4-[[(2S)-2-[[(2S)-2-aminopropanoyl]amino]propanoyl]amino]phenyl]methyl N-[(2R)-2- [[(2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[2-[[4-(1,3-benzothiazol-5-ylamino)-6-tert-butylsulfonyl-7-15 quinolyl]oxy]ethoxy]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy- pyrrolidine-2-carbonyl]amino]-2-[4-(4-methylthiazol-5-yl)phenyl]ethyl]-N-methyl-carbamate Int 201a (15 mg, 9.94 μmol, 53.59% yield, TFA) as yellow solid. LCMS: MS (ESI) m/z 697.1 [1/2M+H]⁺ Step B. Preparation of Int 201b 287 15077.006WO1 To a solution of Int 201a (15 mg, 9.94 μmol, 1 eq, TFA) in DMF (0.1 mL) were added DIEA (3.85 mg, 29.8 μmol, 5.20 μL, 3 eq) and 4-[3-(2,5-dioxopyrrol-1-yl)-2-fluoro-6-[3-oxo-3- (2,3,5,6-tetrafluorophenoxy)propyl]phenoxy]butanoic acid (5.1 mg, 9.94 μmol, 1 eq). The 5 reaction mixture was stirred at 20°C for 1 hr. The pH of mixture was adjusted to 6 with TFA, the residue was poured into ice-water (w/w = 1/1) (1 mL) and stirred for 5 min. The aqueous phase was extracted with DCM/i-PrOH=3/1 (1 mL x 3), aqueous phase was dried with anhydrous Na₂SO₄, filtered and concentrated in vacuum. Afford 4-[6-[3-[[(1S)-2-[[(1S)-2-[4- [[[(2R)-2-[[(2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[2-[[4-(1,3-benzothiazol-5-ylamino)-6-tert-10 butylsulfonyl-7-quinolyl]oxy]ethoxy]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethyl- butanoyl]-4-hydroxy-pyrrolidine-2-carbonyl]amino]-2-[4-(4-methylthiazol-5-yl)phenyl]ethyl]- methyl-carbamoyl]oxymethyl]anilino]-1-methyl-2-oxo-ethyl]amino]-1-methyl-2-oxo- ethyl]amino]-3-oxo-propyl]-3-(2,5-dioxopyrrol-1-yl)-2-fluoro-phenoxy]butanoic acid Int 201b (14 mg, 8.04 μmol, 80.83% yield) as yellow oil. LCMS: MS (ESI) m/z 1741.2 [M+H]⁺ 15 Step C. Preparation of Int 201c 288 15077.006WO1 To a solution of Int 201b (14 mg, 8.04 μmol, 1 eq) in DCM (0.1 mL) were added 2,3,5,6-tetrafluorophenol (1.33 mg, 8.04 μmol, 1 eq), EDCI (7.70 mg, 40.18 μmol, 5 eq). The reaction mixture was stirred at 20 °C for 1hr. The residue was poured into ice-water (w/w = 1/1) 5 (1 mL) and stirred for 5 min. The aqueous phase was extracted with MTBE(1 mL*3) ,then the water phase was extracted with DCM/i-PrOH=3/1 (1 mL*3),the combined organic phase was dried with anhydrous Na2SO4, filtered and concentrated in vacuum to afford (2,3,5,6- tetrafluorophenyl) 4-[6-[3-[[(1S)-2-[[(1S)-2-[4-[[[(2R)-2-[[(2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[2- [[4-(1,3-benzothiazol-5-ylamino)-6-tert-butylsulfonyl-7-10 quinolyl]oxy]ethoxy]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-4-hydroxy- pyrrolidine-2-carbonyl]amino]-2-[4-(4-methylthiazol-5-yl)phenyl]ethyl]-methyl- carbamoyl]oxymethyl]anilino]-1-methyl-2-oxo-ethyl]amino]-1-methyl-2-oxo-ethyl]amino]-3- oxo-propyl]-3-(2,5-dioxopyrrol-1-yl)-2-fluoro-phenoxy]butanoate Int 201c (15 mg, crude) as yellow oil. LCMS: MS (ESI) m/z 945.7 [M/2+H]⁺ 15 Step D. Preparation of RVL-1 289 15077.006WO1 To a solution of Int 201c (15 mg, 7.94 μmol, 1 eq) in DMF (0.1 mL) were added DIEA (3.08 mg, 23.8 μmol, 4.15 μL, 3 eq) and 5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59-nonadecamethyl- 5 4,7,10,13,16,19,22,25,28,31,34,37,40,43,46,49,52,55,58-nonadecaoxo- 2,5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59-icosaazahenhexacontan-61-oic acid Int K (16 mg, 11.1 μmol, 1.4 eq). The reaction mixture was stirred at 20°C for 1 hr. The pH of mixture was adjusted to 6 with TFA, then filtered. The residue was purified by prep-HPLC (column: Phenomenex Luna C1875*30mm*3um;mobile phase: [H₂O(0.1% TFA)- 10 ACN];gradient:15%-45% B over 8.0 min) to give RVL-1 (16 mg, 4.78 μmol, 60.28% yield, 98% purity, TFA) as yellow solid.¹H NMR (400 MHz, DMSO-d6) δ11.58 (s, 1H), 9.92 (s, 1H), 9.54 (s, 1H), 9.27 (s, 1H), 8.99 (d, J = 1.6 Hz, 1H), 8.57-8.35 (m, 2H), 8.22 (d, J = 1.6 Hz, 1H), 8.13 (t, J = 7.2 Hz, 2H), 7.60 (d, J = 8.4 Hz, 3H), 7.51 (s, 1H), 7.45-7.33 (m, 4H), 7.31-7.20 (m, 4H), 7.16-7.00 (m, 2H), 6.82 (d, J = 7.2 Hz, 1H), 5.15-4.71 (m, 4H), 4.59-3.83 (m, 52H), 3.56- 15 3.50 (m, 18H), 3.08-2.69 (m, 62H), 2.45-2.40 (d, J = 8.0 Hz, 3H), 2.09-1.86 (m, 3H), 1.79-1.69 (m, 1H), 1.36 (s, 9H), 1.30 (d, J = 6.8 Hz, 3H), 1.19 (d, J = 6.8 Hz, 3H), 0.93 (s, 9H). LCMS: MS (ESI) m/z 1582.4 [1/2M+H]⁺ Example RVL-2 Synthesis of 64-(6-(3-(((S)-1-(((S)-1-((4-((((((3R,5S)-1-((S)-17- ((4-(benzo[d]thiazol-5-ylamino)-6-(tert-butylsulfonyl)quinolin-7-yl)oxy)-2-(tert-butyl)-4-oxo-20 6,9,12,15-tetraoxa-3-azaheptadecanoyl)-5-((4-(4-methylthiazol-5- yl)benzyl)carbamoyl)pyrrolidin-3-yl)oxy)carbonyl)oxy)methyl)phenyl)amino)-1-oxopropan-2- yl)amino)-1-oxopropan-2-yl)amino)-3-oxopropyl)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2- 290 15077.006WO1 fluorophenoxy)-3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosamethyl- 4,7,10,13,16,19,22,25,28,31,34,37,40,43,46,49,52,55,58,61-icosaoxo- 3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosaazatetrahexacontanoic acid, RVL-2 5 Step A. Preparation of Int 202a To a solution of (2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[2-[[4-(1,3-benzothiazol-5-ylamino)-6- tert-butylsulfonyl-7-quinolyl]oxy]ethoxy]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethyl- butanoyl]-4-hydroxy-N-[[4-(4-methylthiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide, 10 CAS Reg. No.1801547-16-9 (150 mg, 141 μmol, 1 eq) in DMF (2 mL) was added DIEA (54.8 mg, 424 μmol, 73.9 μL, 3 eq) and (4-nitrophenyl) carbonochloridate (57.0 mg, 282 μmol, 2 eq). The mixture was stirred at 20°C for 16 hrs. The reaction mixture was diluted with H2O (5 mL) and extracted with EtOAc (5 mL x 3). The combined organic layers were washed with brine (10 mL) dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue.15 The crude product [(3R,5S)-1-[(2S)-2-[[2-[2-[2-[2-[2-[[4-(1,3-benzothiazol-5-ylamino)-6-tert- butylsulfonyl-7-quinolyl]oxy]ethoxy]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethyl- butanoyl]-5-[[4-(4-methylthiazol-5-yl)phenyl]methylcarbamoyl]pyrrolidin-3-yl] (4- nitrophenyl)methyl carbonate Int 202a (170 mg, crude) (yellow oil) was used into the next step without further purification. LCMS: MS (ESI) m/z 1225.4[M+H]⁺ 20 Step B. Preparation of Int 202b 291 15077.006WO1 To a solution of Int 202a (170 mg, 137 μmol, 1 eq) in DMF (2 mL) was added 9H- fluoren-9-ylmethyl N-[(1S)-2-[[(1S)-2-[4-(hydroxymethyl)anilino]-1-methyl-2-oxo- ethyl]amino]-1-methyl-2-oxo-ethyl]carbamate (134 mg, 274 μmol, 2 eq) DIEA (53.2 mg, 411 5 μmol, 71.7 μL, 3 eq) and DMAP (16.8 mg, 137 μmol, 1 eq). The mixture was stirred at 20°C for 2 hrs. The crude product [(3R,5S)-1-[(2S)-2-[[2-[2-[2-[2-[2-[[4-(1,3-benzothiazol-5-ylamino)-6- tert-butylsulfonyl-7-quinolyl]oxy]ethoxy]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethyl- butanoyl]-5-[[4-(4-methylthiazol-5-yl)phenyl]methylcarbamoyl]pyrrolidin-3-yl] [4-[[(2S)-2- [[(2S)-2-(9H-fluoren-9- 10 ylmethoxycarbonylamino)propanoyl]amino]propanoyl]amino]phenyl]methyl carbonate Int 202b (0.21 g, crude) was used into the next step without further purification. LCMS: MS (ESI) m/z 787.5[M/2+H]⁺ Step C. Preparation of Int 202c 292 15077.006WO1 To a solution of Int 202b (200 mg, 127 μmol, 1 eq) in DMF (2 mL) was added piperidine (54.1 mg, 635 μmol, 62.7 μL, 5 eq). The mixture was stirred at 20°C for 1 hr. The reaction solution was adjusted to PH=7. The residue was purified by prep-HPLC (column: Waters Xbridge BEH C18100 x 30mm x 10um; mobile phase: [H2O (10mM NH4HCO3)-ACN]; 5 gradient:38%-70% B over 8.0 min). Compound [4-[[(2S)-2-[[(2S)-2- aminopropanoyl]amino]propanoyl]amino]phenyl]methyl [(3R,5S)-1-[(2S)-2-[[2-[2-[2-[2-[2-[[4- (1,3-benzothiazol-5-ylamino)-6-tert-butylsulfonyl-7- quinolyl]oxy]ethoxy]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-5-[[4-(4- methylthiazol-5-yl)phenyl]methylcarbamoyl]pyrrolidin-3-yl] carbonate Int 202c (30 mg, 22.2 10 μmol, 17.8% yield) was obtained as a white solid. LCMS: MS (ESI) m/z 676.5[M/2+H]⁺ Step D. Preparation of Int 202d To a solution of Int 202c (20.0 mg, 14.8 μmol, 1 eq) in DMF (0.2 mL) was added DIEA (5.74 mg, 44.3 μmol, 7.73 μL, 3 eq) and 4-[3-(2,5-dioxopyrrol-1-yl)-2-fluoro-6-[3-oxo-3- 15 (2,3,5,6-tetrafluorophenoxy)propyl]phenoxy]butanoic acid (9.12 mg, 17.7 μmol, 1.2 eq). The mixture was stirred at 20°C for 1 hr. The reaction mixture was adjusted to pH=6 with TFA and then diluted with H2O (1 mL) and extracted with DCM/i-PrOH = 3/1(1 mL x 3). The combined organic layers were dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The crude product 4-[6-[3-[[(1S)-2-[[(1S)-2-[4-[[(3R,5S)-1-[(2S)-2-[[2-[2-[2-[2-[2-20 [[4-(1,3-benzothiazol-5-ylamino)-6-tert-butylsulfonyl-7- quinolyl]oxy]ethoxy]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-5-[[4-(4- methylthiazol-5-yl)phenyl]methylcarbamoyl]pyrrolidin-3-yl]oxycarbonyloxymethyl]anilino]-1- 293 15077.006WO1 methyl-2-oxo-ethyl]amino]-1-methyl-2-oxo-ethyl]amino]-3-oxo-propyl]-3-(2,5-dioxopyrrol-1- yl)-2-fluoro-phenoxy]butanoic acid Int 202d (25 mg, crude) was used into the next step without further purification. LCMS: MS (ESI) m/z 850.1[M/2+H]⁺ Step E. Preparation of Int 202e 5 To a solution of Int 202d (25.0 mg, 14.7 μmol, 1 eq) in DCM (0.5 mL) was added EDCI (11.2 mg, 58.8 μmol, 4 eq) and 2,3,5,6-tetrafluorophenol (7.33 mg, 44.15 μmol, 3 eq). The mixture was stirred at 20°C for 1 hr. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: Phenomenex Luna 10 C1875 x 30mm x 3um; mobile phase: [H₂O(0.1% TFA)-ACN];gradient:35%-65% B over 8.0 min). Compound (2,3,5,6-tetrafluorophenyl) 4-[6-[3-[[(1S)-2-[[(1S)-2-[4-[[(3R,5S)-1-[(2S)-2- [[2-[2-[2-[2-[2-[[4-(1,3-benzothiazol-5-ylamino)-6-tert-butylsulfonyl-7- quinolyl]oxy]ethoxy]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethyl-butanoyl]-5-[[4-(4- methylthiazol-5-yl)phenyl]methylcarbamoyl]pyrrolidin-3-yl]oxycarbonyloxymethyl]anilino]-1-15 methyl-2-oxo-ethyl]amino]-1-methyl-2-oxo-ethyl]amino]-3-oxo-propyl]-3-(2,5-dioxopyrrol-1- yl)-2-fluoro-phenoxy]butanoate Int 202e (16 mg, 8.66 μmol, 58.8% yield) was obtained as a white solid.¹H NMR: (400 MHz, DMSO-d6) δ11.57 (s, 1H), 9.95 (s, 1H), 9.54 (s, 1H), 9.27 (s, 1H), 9.04-8.87 (m, 1H), 8.68-8.60 (m, 1H), 8.48 (d, J = 6.8 Hz, 1H), 8.39 (d, J = 8.4 Hz, 1H), 8.22 (d, J = 1.6 Hz, 1H), 8.13 (d, J = 7.2 Hz, 2H), 7.99-7.87 (m, 1H), 7.71-7.57 (m, 3H), 7.50 (s, 20 1H), 7.47-7.23 (m, 9H), 7.20-7.02 (m, 2H), 6.82 (d, J = 7.4 Hz, 1H), 5.24 (s, 1H), 5.09-5.06 (m, 2H), 4.57-4.25 (m, 8H), 4.15-3.81 (m, 8H), 3.67-3.49 (m, 16H), 3.02 (t, J = 7.2 Hz, 2H),2.93- 294 15077.006WO1 2.85(m, 2H), 2.45 (s, 3H), 2.17-2.03 (m, 2H), 1.43-1.16 (m, 15H), 0.96 (s, 9H). LCMS: MS (ESI) m/z 924.4[M/2+H]⁺ Step F. Preparation of RVL-2 5 To a solution of Int 202e (15 mg, 7.65 μmol, 1 eq, TFA) in DMF (0.1 mL) were added DIEA (2.97 mg, 22.0 μmol, 4.00 μL, 3 eq) and 5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59-nonadecamethyl- 4,7,10,13,16,19,22,25,28,31,34,37,40,43,46,49,52,55,58-nonadecaoxo- 2,5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59-icosaazahenhexacontan-61-oic acid 10 Int K (14.3 mg, 9.94 μmol, 1.3 eq). The reaction mixture was stirred at 20°C for 1 hr. The pH of mixture was adjusted to 6 with TFA, then was filtered. The residue was purified by prep- HPLC (column: Phenomenex Luna C1875*30mm*3um;mobile phase: [H₂O(0.1% TFA)- ACN];gradient:20%-50% B over 8.0 min) to afford RVL-2 (20 mg, 6.06 μmol, 79.22% yield, 98% purity, TFA) as white solid.¹H NMR: (400 MHz, DMSO-d6) δ11.58 (s, 1H), 9.96 (d, J = 15 1.6 Hz, 1H), 9.54 (s, 1H), 9.27 (s, 1H), 8.98 (s, 1H), 8.63 (t, J = 6.0 Hz, 1H), 8.48 (d, J = 7.2 Hz, 1H), 8.39 (d, J = 8.4 Hz, 1H), 8.23 (d, J = 2.0 Hz, 1H), 8.15 (t, J = 8.4 Hz, 2H), 7.66-7.58 (m, 3H), 7.50 (s, 1H), 7.46-7.41 (m, 1H), 7.40 (s, 3H), 7.33 (d, J = 8.8 Hz, 2H), 7.25 (s, 2H), 7.14- 6.99 (m, 2H), 6.82 (d, J = 7.2 Hz, 1H), 5.24 (d, J = 1.2 Hz, 1H), 5.08 (d, J = 2.0 Hz, 2H), 4.53- 3.82 (m, 55H), 3.64-3.52 (m, 18H), 3.05-2.70 (m, 60H), 2.43 (s, 3H), 2.20-2.06 (m, 1H), 1.99- 20 1.82 (m, 2H), 1.39-1.33 (m, 9H), 1.30 (d, J = 7.2 Hz, 3H), 1.19 (d, J = 7.2 Hz, 3H), 0.99-0.89 (m, 9H). LCMS: MS (ESI) m/z 3117.32 [M-H]- 295 15077.006WO1 Example RVL-3 Synthesis of 64-(6-(3-(((S)-1-(((S)-1-((4-((2-((S)-1-((2S,4R)-1- ((S)-17-((4-(benzo[d]thiazol-5-ylamino)-6-(tert-butylsulfonyl)quinolin-7-yl)oxy)-2-(tert-butyl)- 4-oxo-6,9,12,15-tetraoxa-3-azaheptadecanoyl)-4-hydroxypyrrolidine-2-carboxamido)ethyl)-5- (4-methylthiazol-5-yl)phenoxy)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-1-oxopropan- 5 2-yl)amino)-3-oxopropyl)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-fluorophenoxy)- 3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosamethyl- 4,7,10,13,16,19,22,25,28,31,34,37,40,43,46,49,52,55,58,61-icosaoxo- 3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosaazatetrahexacontanoic acid, RVL-3 10 Step A. Preparation of Int 203a. To a mixture of (1S)-1-[2-benzyloxy-4-(4-methylthiazol-5-yl)phenyl]ethanamine (5 g, 13.8 mmol, 1.0 eq, HCl) and DIEA (12.5 g, 96.9 mmol, 16.8 mL, 7.0 eq) in DMF (70 mL) were15 added(2S,4R)-1-[(2S)-2-(tert-butoxycarbonylamino)-3,3-dimethylbutanoyl]-4-hydroxy- pyrrolidine-2-carboxylic acid (4.77 g, 13.8 mmol, 1.0 eq), HOBt (2.81 g, 20.7 mmol, 1.5 eq) and EDCI (3.98 g, 20.7 mmol, 1.5 eq) at 0°C. The mixture was warmed to 25°C and stirred for 1hr. The reaction mixture was quenched by addition H2O (30 mL), and then extracted with EtOAc 296 15077.006WO1 (35 mL x 3). The combined organic layers were washed with brine 15 mL, dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0~80% Ethyl acetate/Petroleum ethergradient @ 80 mL/min) to give tert-butyl N-[(1S)-1- 5 [(2S,4R)-2-[[(1S)-1-[2-benzyloxy-4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]carbamate, Int 203a (6.6 g, 10.14 mmol, 73.2% yield) as a white solid. LCMS: MS (ESI) m/z 651.2[M+H]⁺ Step B. Preparation of Int 203b. 10 To a solution of Int 203a (5.5 g, 8.45 mmol, 1.0 eq) in DCM (60 mL) was added BBr₃ (2 M, 12.6 mL, 3.0 eq) dropwise at -78°C. The reaction mixture was stirred at -78°C for 1hr. The reaction mixture was warmed to 0°C and quenched by addition water 10 mL and aq. NaHCO₃ until pH = 8. The reaction mixture was filtered and the filter cake was washed with 30 mL of DCM, dried in vacuum to give crude (2S,4R)-1-[(2S)-2-amino-3,3-dimethyl-butanoyl]-4-15 hydroxy-N-[(1S)-1-[2-hydroxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2- carboxamide, Int 203b (5 g, crude) was obtained as green solid and used into the next step without further purification. LCMS: MS (ESI) m/z 461.1[M+H]⁺ Step C. Preparation of Int 203c. To a solution of Int 203b (4 g, 8.68 mmol, 1.0 eq) in THF (40 mL) was added Et₃N (1.32 20 g, 13.0 mmol, 1.81 mL, 1.5 eq) and Boc₂O (2.27 g, 10.4 mmol, 2.39 mL, 1.2 eq). The reaction mixture was stirred at 25 °C for 1hr. The reaction mixture was quenched by addition H2O (20 mL), and then extracted with EtOAc (20 mL x 3). The combined organic layers were washed with brine (15 mL x 2), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g 25 SepaFlash® Silica Flash Column, Eluent of 0~10% Ethylacetate/MeOH @ 70 mL/min) to give tert-butyl N-[(1S)-1-[(2S,4R)-4-hydroxy-2-[[(1S)-1-[2-hydroxy-4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]carbamate Int.203c (2.9 g, 5.17 mmol, 59.55% yield) as yellow oil. LCMS: MS (ESI) m/z 561.4[M+H]⁺ Step D. Preparation of Int 203d. 30 To a mixture of Int.203c (163 mg, 291 μmol, 1.0 eq) and allyl N-[(1S)-2-[[(1S)-2-[4- (chloromethyl) anilino]-1-methyl-2-oxo-ethyl]amino]-1-methyl-2-oxo-ethyl]carbamate (0.15 g, 408 μmol, 1.4 eq) in DMF (3 mL) was added K2CO3 (121 mg, 874 μmol, 3.0 eq) in one portion at 20°C and stirred at 20°C for 12 hrs. The residue was poured into ice-water (w/w = 1/1) (20 mL) and stirred for 2 min. The aqueous phase was extracted with ethyl acetate (10 mL x 3). 35 The combined organic phase was washed with brine (10 mL x 2), dried with anhydrous Na2SO4, 297 15077.006WO1 filtered and concentrated in vacuum. Compound tert-butyl N-[(1S)-1-[(2S,4R)-2-[[(1S)-1-[2- [[4-[[(2S)-2-[[(2S)-2-(allyloxycarbonylamino)propanoyl]amino]propanoyl] amino]phenyl]methoxy]-4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]-4-hydroxy- pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]carbamate Int.203d (0.18 g, crude) was obtained 5 as yellow oil. LCMS: MS (ESI) m/z 892.5 [M+H]⁺ Step E. Preparation of Int 203e. To a mixture of Int.203d (0.15 g, 168 μmol, 1.0 eq) in EtOAc (2 mL) was added HCl/EtOAc (4 M, 4 mL, 95.2 eq) in one portion at 20°C and stirred at 20°C for 1 hr. The10 mixture was concentrated. Compound allyl N-[(1S)-2-[[(1S)-2-[4-[[2-[(1S)-1-[[(2S,4R)-1- [(2S)-2-amino-3,3-dimethyl-butanoyl]-4-hydroxy-pyrrolidine-2-carbonyl]amino]ethyl]-5-(4- methylthiazol-5-yl)phenoxy]methyl]anilino]-1-methyl-2-oxo-ethyl]amino]-1-methyl-2-oxo- ethyl]carbamate Int.203e (0.12 g, crude, HCl) was obtained as a yellow solid. LCMS: MS (ESI) m/z 792.4 [M+H]⁺ 15 Step F: Preparation of Int 203f. 298 15077.006WO1 To a mixture of 2-[2-[2-[2-[2-[[4-(1,3-benzothiazol-5-ylamino)-6-tert-butylsulfonyl-7- quinolyl]oxy]ethoxy]ethoxy]ethoxy]ethoxy]acetic acid (65.2 mg, 101 μmol, 1.0 eq) in DMF (2 mL) was added HATU (45.9 mg, 121 μmol, 1.2 eq), DIEA (65.0 mg, 503 μmol, 87.6 μL, 5.0 eq) 5 and Int.203e (0.1 g, 121 μmol, 1.2 eq, HCl) in one portion at 20°C and stirred at 20°C for 1 hr. The residue was poured into ice-water (w/w = 1/1) (20 mL). The aqueous phase was extracted with ethyl acetate (10 mL x 3). The combined organic phase was washed with brine (10 mL x 2), dried with anhydrous Na₂SO₄, filtered and concentrated in vacuum. Compound allyl N- [(1S)-2-[[(1S)-2-[4-[[2-[(1S)-1-[[(2S,4R)-1-[(2S)-2-[[2-[2-[2-[2-[2-[[4-(1,3-benzothiazol-5-10 ylamino)-6-tert-butylsulfonyl-7-quinolyl]oxy]ethoxy]ethoxy] ethoxy]ethoxy]acetyl]amino]-3,3- dimethyl-butanoyl]-4-hydroxy-pyrrolidine-2-carbonyl] amino]ethyl]-5-(4-methylthiazol-5- yl)phenoxy]methyl]anilino]-1-methyl-2-oxo-ethyl] amino]-1-methyl-2-oxo-ethyl]carbamate Int. 203f (0.16 g, crude) was obtained as yellow oil. LCMS: MS (ESI) m/z 711.8 [M/2+H]⁺ Step G. Preparation of Int 203g. 299 15077.006WO1 To a mixture of Int.203f (0.15 g, 106 μmol, 1.0 eq) in DCM (3 mL) was added DABCO (94.7 mg, 844 μmol, 92.8 μL, 8.0 eq) and Pd(PPh₃)₄ (36.6 mg, 31.6 μmol, 0.3 eq) in one portion at 20°C under N2. The mixture was stirred at 20°C for 0.5 hr. The mixture was quenched with 5 TFA until pH = 6, and filtered and purified by prep-HPLC(column: Phenomenex Luna C18 75*30mm*3um;mobile phase: [H2O(0.1% TFA)-ACN];gradient:15%-45% B over 8.0 min) to give (2S,4R)-N-[(1S)-1-[2-[[4-[[(2S)-2-[[(2S)-2- aminopropanoyl]amino]propanoyl]amino]phenyl]methoxy]-4-(4-methylthiazol-5- yl)phenyl]ethyl]-1-[(2S)-2-[[2-[2-[2-[2-[2-[[4-(1,3-benzothiazol-5-ylamino)-6-tert-10 butylsulfonyl-7-quinolyl]oxy]ethoxy]ethoxy]ethoxy]ethoxy]acetyl]amino]-3,3-dimethyl- butanoyl]-4-hydroxy-pyrrolidine-2-carboxamide Int.203g (45 mg, 33.64 μmol, 31.89% yield) as yellow solid. LCMS: MS (ESI) m/z 669.3 [M/2+H]⁺ Step H: Preparation of RVL-3 300 15077.006WO1 To a mixture of Int.203g (18 mg, 11.50 μmol, 1.0 eq, 2TFA) in DMF (0.5 mL) were added DIEA (7.43 mg, 57.5 μmol, 10.0 μL, 5.0 eq) and 64-(6-(2-carboxyethyl)-3-(2,5-dioxo-2,5- dihydro-1H-pyrrol-1-yl)-2-fluorophenoxy)- 5 3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosamethyl- 4,7,10,13,16,19,22,25,28,31,34,37,40,43,46,49,52,55,58,61-icosaoxo- 3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosaazatetrahexacontanoic acid Int L (15.57 mg, 8.05 μmol, 0.7 eq) in one portion at 20°C and stirred at 20°C for 0.5 hr. The mixture was added TFA to adjust pH between 6 and 7 and filtered and further purification by10 prep-HPLC(column: Welchrom CSH C18100*30*7;mobile phase: [H2O(0.1%TFA)- ACN];gradient:20%-40% B over 15.0 min) to give RVL-3 (9 mg, 2.90 μmol, 25.20% yield) as yellow solid.¹H NMR (400 MHz, DMSO-d₆) δ = 9.99-9.89 (m, 1H), 9.48-9.42 (m, 1H), 8.94 (s, 1H), 8.45 (d, J = 6.0 Hz, 1H), 8.37 (dd, J = 1.6, 9.6 Hz, 1H), 8.31-8.21 (m, 1H), 8.18-8.05 (m, 2H), 7.61 (d, J = 8.0 Hz, 2H), 7.57-7.50 (m, 1H), 7.49-7.44 (m, 1H), 7.40 (d, J = 8.4 Hz, 2H), 15 7.28 (d, J = 7.6 Hz, 1H), 7.23-7.17 (m, 2H), 7.14-7.07 (m, 1H), 7.07-6.97 (m, 3H), 6.88-6.82 (m, 1H), 5.24-5.17 (m, 1H), 5.17-5.12 (m, 2H), 4.55-4.49 (m, 1H), 4.48-4.42 (m, 1H), 4.36-4.17 (m, 24H), 4.12-3.89 (m, 24H), 3.83 (d, J = 4.4 Hz, 2H), 3.63-3.52 (m, 16H), 2.96-2.70 (m, 64H), 2.37 (s, 3H), 2.09-2.03 (m, 1H), 2.03-1.85 (m, 4H), 1.81-1.70 (m, 1H), 1.35-1.32 (m, 9H), 1.30 (t, J = 7.6 Hz, 6H), 1.18 (d, J = 7.2 Hz, 3H), 0.91 (s, 9H). LCMS: MS (ESI) m/z 3103.37 [M-H]- 301 15077.006WO1 Example KVL-7 Synthesis of 64-(6-(3-(((2S)-1-(((2S)-1-((4-((((2-((2S,4R)-1-((S)- 2-(4-(4-((S)-4-(4-(5-((S)-2-amino-3-cyano-4-methyl-4,5,6,7-tetrahydrobenzo[b]thiophen-4-yl)- 1,2,4-oxadiazol-3-yl)pyrimidin-2-yl)-3-methyl-1,4-diazepan-1-yl)butoxy)-1H-1,2,3-triazol-1- yl)-3-methylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)-2-(4-(4-methylthiazol-5- 5 yl)phenyl)ethyl)(methyl)carbamoyl)oxy)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-1- oxopropan-2-yl)amino)-3-oxopropyl)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2- fluorophenoxy)-3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosamethyl- 4,7,10,13,16,19,22,25,28,31,34,37,40,43,46,49,52,55,58,61-icosaoxo- 3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosaazatetrahexacontanoic acid, 10 KVL-7 To a mixture of (2S,4R)-1-[(2S)-2-[4-[4-[(3S)-4-[4-[5-[(4S)-2-amino-3-cyano-4-methyl- 6,7-dihydro-5H-benzothiophen-4-yl]-1,2,4-oxadiazol-3-yl]pyrimidin-2-yl]-3-methyl-1,4-15 diazepan-1-yl]butoxy]triazol-1-yl]-3-methyl-butanoyl]-4-hydroxy-N-[(1R)-2-(methylamino) -1- [4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide, KV-13 (0.05 g, 46.8 μmol, 1.0 eq, HCl) in DMF (2 mL) were added DIEA (30.2 mg, 234 μmol, 40.7 μL, 5.0 eq) and [4- 302 15077.006WO1 [[(2S)-2-[[(2S)-2-(9H-fluoren-9-ylmethoxycarbonylamino)propanoyl]amino]propanoyl] amino]phenyl]methyl (4-nitrophenyl) carbonate (30.5 mg, 46.8 μmol, 1.0 eq) in one portion at 20°C and stirred for 20 min, then the mixture solution was added piperidine (19.92 mg, 234 μmol, 23.1 μL, 5.0 eq) and stirred at 20°C for 10 min. The mixture was puriifed by prep- 5 HPLC(column: Phenomenex Luna C1875*30mm*3um;mobile phase: [H2O(0.1% TFA)- ACN];gradient:20%-45% B over 8.0 min) to give [4-[[(2S)-2-[[(2S)-2- aminopropanoyl]amino]propanoyl]amino]phenyl]methyl N-[(2R)-2-[[(2S,4R)-1-[(2S)-2-[4-[4- [(3S)-4-[4-[5-[(4S)-2-amino-3-cyano-4-methyl-6,7-dihydro-5H-benzothiophen-4-yl]-1,2,4- oxadiazol-3-yl]pyrimidin-2-yl]-3-methyl-1,4-diazepan-1-yl]butoxy]triazol-1-yl]-3-methyl-10 butanoyl]-4-hydroxy-pyrrolidine-2-carbonyl]amino]-2-[4-(4-methylthiazol-5-yl)phenyl]ethyl]- N-methyl-carbamate Int 204a (19 mg, 14.35 μmol, 30.68% yield) as yellow solid. LCMS: MS (ESI) m/z 1322.6 [M+H]⁺ Step B. Preparation of Int 204b 15 To a mixture of Int 204a (10 mg, 7.56 μmol, 1.0 eq) and 4-[3-(2,5-dioxopyrrol-1-yl)-2- fluoro-6-[3-oxo-3-(2,3,5,6-tetrafluorophenoxy) propyl]phenoxy]butanoic acid (3.88 mg, 7.56 303 15077.006WO1 μmol, 1.0 eq) in DMF (0.5 mL) was added DIEA (2.93 mg, 22.7 μmol, 3.95 μL, 3.0 eq) in one portion at 20°C and stirred at 20°C for 0.5 hr. The mixture was diluted with water and extracted with DCM:i-PrOH=3:1(5 mL x 3). The organic layer was washed with brine, dried over Na2SO4, filtered and concentrated. Compound 4-[6-[3-[[(1S)-2-[[(1S)-2-[4-[[[(2R)-2-[[(2S,4R)- 5 1-[(2S)-2-[4-[4-[(3S)-4-[4-[5-[(4S)-2-amino-3-cyano-4-methyl-6,7-dihydro-5H-benzothiophen- 4-yl]-1,2,4-oxadiazol-3-yl]pyrimidin-2-yl]-3-methyl-1,4-diazepan-1-yl]butoxy]triazol-1-yl]-3- methyl-butanoyl]-4-hydroxy-pyrrolidine-2-carbonyl]amino]-2-[4-(4-methylthiazol-5- yl)phenyl]ethyl]-methyl-carbamoyl]oxymethyl]anilino]-1-methyl-2-oxo-ethyl]amino]-1-methyl- 2-oxo-ethyl]amino]-3-oxo-propyl]-3-(2,5-dioxopyrrol-1-yl)-2-fluoro-phenoxy]butanoic acid Int 10 204b (12 mg, crude) was obtained as yellow oil. LCMS: MS (ESI) m/z 836.4 [M/2+H]⁺ Step C. Preparation of Int 204c To a mixture of Int 204b (12 mg, 7.18 μmol, 1.0 eq) and 2,3,5,6-tetrafluorophenol (3.58 mg, 21.6 μmol, 3.0 eq) in DCM (1 mL) was added EDCI (6.88 mg, 35.9 μmol, 5.0 eq) in one 15 portion at 20°C and stirred at 20°C for 0.5 hr. The mixture was purified by prep-HPLC(column: Phenomenex Luna C1875*30mm*3um;mobile phase: [H2O(0.1% TFA)-ACN];gradient:30%- 60% B over 8.0 min) to give (2,3,5,6-tetrafluorophenyl) 4-[6-[3-[[(1S)-2-[[(1S)-2-[4-[[[(2R)-2- [[(2S,4R)-1-[(2S)-2-[4-[4-[(3S)-4-[4-[5-[(4S)-2-amino-3-cyano-4-methyl-6,7-dihydro-5H- benzothiophen-4-yl] -1,2,4-oxadiazol-3-yl]pyrimidin-2-yl]-3-methyl-1,4-diazepan-1-20 yl]butoxy]triazol-1-yl]-3-methyl-butanoyl]-4-hydroxy-pyrrolidine-2-carbonyl]amino]-2-[4-(4- methylthiazol-5-yl) phenyl]ethyl]-methyl-carbamoyl]oxymethyl]anilino]-1-methyl-2-oxo- ethyl]amino]-1-methyl-2-oxo-ethyl]amino]-3-oxo-propyl]-3-(2,5-dioxopyrrol-1-yl)-2-fluoro- phenoxy] butanoate Int 204c (5 mg, 2.75 μmol, 38.28% yield) as yellow solid. LCMS: MS (ESI) m/z 910.4 [M/2+H]⁺ 25 Step D. Preparation of KVL-7 304 15077.006WO1 To a mixture of Int 204c (5 mg, 2.75 μmol, 1.0 eq) and 5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59-nonadecamethyl- 4,7,10,13,16,19,22,25,28,31,34,37,40,43,46,49,52,55,58-nonadecaoxo- 5 2,5,8,11,14,17,20,23,26,29,32,35,38,41,44,47,50,53,56,59-icosaazahenhexacontan-61-oic acid Int K (3.96 mg, 2.75 μmol, 1.0 eq) in DMF (0.5 mL) was added DIEA (1.78 mg, 13.7 μmol, 2.39 μL, 5.0 eq) in one portion at 20°C and stirred at 20°C for 0.5 hr. The mixture was purified by prep-HPLC(column: Phenomenex Luna C1875*30mm*3um;mobile phase: [H2O(0.1% TFA)-ACN];gradient:20%-50% B over 8.0 min) to give KVL-7 (5.5 mg, 1.78 μmol, 64.70% 10 yield) as white solid.¹H NMR: (400 MHz, DMSO-d₆) δ = 10.01-9.82 (m, 1H), 9.42-9.21 (m, 1H), 9.05-8.90 (m, 1H), 8.71-8.45 (m, 1H), 8.15-8.10 (m, 2H), 7.65-7.53 (m, 2H), 7.49-7.31 (m, 1H), 7.30-7.15 (m, 6H), 7.14-7.06 (m, 3H), 7.06-6.99 (m, 1H), 5.29-4.69 (m, 5H), 4.48-4.16 (m, 21H), 4.14-3.88 (m, 21H), 3.83-3.44 (m, 5H), 3.27-3.13 (m, 6H), 3.03-2.68 (m, 60H), 2.43 (d, J = 8.0 Hz, 8H), 2.13-2.01 (m, 3H), 1.99-1.82 (m, 9H), 1.80-1.63 (m, 7H), 1.36-1.07 (m, 12H), 15 1.06-0.97 (m, 3H), 0.66 (s, 3H). LCMS: MS (ESI) m/z 3089.41[M-H]- Example KVL-10 Synthesis of 64-(6-(3-(((S)-1-(((S)-1-((4-((((3-(5-(4-((R)-1- ((2S,4R)-1-((S)-2-(4-(4-((S)-4-(4-(5-((S)-2-amino-3-cyano-4-methyl-4,5,6,7- tetrahydrobenzo[b]thiophen-4-yl)-1,2,4-oxadiazol-3-yl)pyrimidin-2-yl)-3-methyl-1,4-diazepan- 1-yl)butoxy)-1H-1,2,3-triazol-1-yl)-3-methylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)-2-20 hydroxyethyl)phenyl)thiazol-4-yl)prop-2-yn-1- yl)(methyl)carbamoyl)oxy)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-1-oxopropan-2- yl)amino)-3-oxopropyl)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-fluorophenoxy)- 3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosamethyl- 4,7,10,13,16,19,22,25,28,31,34,37,40,43,46,49,52,55,58,61-icosaoxo- 305 15077.006WO1 3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosaazatetrahexacontanoic acid, KVL-10 5 To a solution of (2S,4R)-1-[(2S)-2-[4-[4-[(3S)-4-[4-[5-[(4S)-2-amino-3-cyano-4-methyl- 6,7-dihydro-5H-benzothiophen-4-yl]-1,2,4-oxadiazol-3-yl]pyrimidin-2-yl]-3-methyl-1,4- diazepan-1-yl]butoxy]triazol-1-yl]-3-methyl-butanoyl]-4-hydroxy-N-[(1R)-2-hydroxy-1-[4-[4- [3-(methylamino)prop-1-ynyl]thiazol-5-yl]phenyl]ethyl]pyrrolidine-2-carboxamide, KV-17 (30 mg, 27.9 μmol, 1 eq) in DMF (1 mL) was added DIEA (18.1 mg, 139 μmol, 24.4 μL, 5 eq) and10 [4-[[(2S)-2-[[(2S)-2-(9H-fluoren-9- ylmethoxycarbonylamino)propanoyl]amino]propanoyl]amino]phenyl]methyl (4-nitrophenyl) carbonate (27.40 mg, 41.9 μmol, 1.5 eq). The mixture was stirred at 20°C for 1 hr. then piperidine (12.1 mg, 141 μmol, 14.0 μL, 5 eq) was added. The mixture was stirred at 20°C for 1 hr. The reaction solution was adjusted to pH = 7 with TFA. The residue was purified by prep- 15 HPLC (column: WePure Biotech XP tC18150 x 40 x 70um; mobile phase: [H2O(10mM NH4HCO3)-ACN];gradient:40%-68% B over 8.0 min). Compound [4-[[(2S)-2-[[(2S)-2- aminopropanoyl]amino]propanoyl]amino]phenyl]methyl N-[3-[5-[4-[(1R)-1-[[(2S,4R)-1-[(2S)- 2-[4-[4-[(3S)-4-[4-[5-[(4S)-2-amino-3-cyano-4-methyl-6,7-dihydro-5H-benzothiophen-4-yl]- 1,2,4-oxadiazol-3-yl]pyrimidin-2-yl]-3-methyl-1,4-diazepan-1-yl]butoxy]triazol-1-yl]-3-methyl-20 butanoyl]-4-hydroxy-pyrrolidine-2-carbonyl]amino]-2-hydroxy-ethyl]phenyl]thiazol-4-yl]prop- 2-ynyl]-N-methyl-carbamate Int 205a (15 mg, 11.0 μmol, 38.8% yield) was obtained as a white solid.¹H NMR (400 MHz, DMSO-d6) δ10.05 (s, 1H), 9.12-9.01 (m, 1H), 8.63-8.54 (m, 1H), 8.49 (d, J = 8.0 Hz, 1H), 7.75 (s, 2H), 7.60-7.47 (m, 2H), 7.43-7.27 (m, 4H), 7.16-6.94 (m, 3H), 5.20-5.01 (m, 4H), 4.93-4.79 (m, 2H), 4.77-4.72 (m, 1H), 4.51-4.40 (m, 2H), 4.37-4.21 (m, 1H), 25 4.04 (t, J = 6.4 Hz, 2H), 3.82-3.50 (m, 4H), 3.23-3.08 (m, 2H), 3.06-2.79 (m, 5H), 2.19-2.05 (m, 306 15077.006WO1 2H), 1.99-1.83 (m, 4H), 1.82-1.46 (m, 11H), 1.30 (d, J = 6.8 Hz, 3H), 1.13 (d, J = 6.8 Hz, 3H), 1.05-10.01 (m, 6H), 0.66 (d, J = 6.4 Hz, 3H). LCMS (ESI) m/z 682.5 [M/2+H]⁺ Step B. Preparation of KVL-10 5 To a solution of Int 205a (11 mg, 8.07 μmol, 1 eq) in DMF (0.1 mL) was added DIEA (3.13 mg, 24.2 μmol, 4.22 μL, 3 eq) and 64-(3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-fluoro- 6-(3-oxo-3-(2,3,5,6-tetrafluorophenoxy)propyl)phenoxy)- 3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosamethyl- 4,7,10,13,16,19,22,25,28,31,34,37,40,43,46,49,52,55,58,61-icosaoxo-10 3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosaazatetrahexacontanoic acid Int- L (15.6 mg, 8.07 μmol, 1 eq). The mixture was stirred at 20°C for 1 hr. The reaction solution was adjusted to pH = 6 with TFA. The residue was purified by prep-HPLC (column: Phenomenex Luna C₁₈75 x 30mm x 3um; mobile phase: [H₂O(0.1% TFA)-ACN];gradient:15%- 45% B over 8.0 min) to give KVL-10 (5 mg, 1.60 μmol, 19.8% yield) was obtained as a white 15 solid.¹ H NMR (400 MHz, DMSO-d6) δ9.93-9.87 (m, 1H), 9.34-9.28 (m, 1H), 9.05 (s, 1H), 8.65 (d, J = 3.2 Hz, 1H), 8.47 (d, J = 7.2 Hz, 1H), 8.11 (t, J = 5.6 Hz, 2H), 7.78-7.71 (m, 2H), 7.69 (s, 1H), 7.63-7.52 (m, 2H), 7.36 (d, J = 8.0 Hz, 2H), 7.33-7.25 (m, 3H), 7.24 (s, 2H), 7.13-7.01 (m, 4H), 5.11 (d, J = 10.4 Hz, 1H), 5.04 (s, 2H), 4.89-4.79 (m, 2H), 4.38-3.93 (m, 50H), 3.76-3.67 (m, 2H), 3.64-3.56 (m, 2H), 2.96-2.71 (m, 70H), 2.11-2.03 (m, 2H), 1.97-1.81 (m, 10H), 1.78 (s, 20 4H), 1.76-1.69 (m, 3H), 1.32-1.21 (m, 6H), 1.19-1.18 (m, 3H), 1.15-1.11 (m, 4H), 1.02 (d, J = 6.8 Hz, 3H), 0.66 (d, J = 6.8 Hz, 3H). LCMS (ESI) m/z 3129.3 [M-H]- 307 15077.006WO1 Example KVL-11 Synthesis of 64-(6-(3-(((S)-1-(((S)-1-((4-((6-((S)-1-((2S,4R)-1- ((S)-2-(4-(4-((S)-4-(4-(5-((S)-2-amino-3-cyano-4-methyl-4,5,6,7-tetrahydrobenzo[b]thiophen-4- yl)-1,2,4-oxadiazol-3-yl)pyrimidin-2-yl)-3-methyl-1,4-diazepan-1-yl)butoxy)-1H-1,2,3-triazol- 1-yl)-3-methylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)ethyl)-2-fluoro-3-(4- 5 methylthiazol-5-yl)phenoxy)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-1-oxopropan-2- yl)amino)-3-oxopropyl)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-fluorophenoxy)- 3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosamethyl- 4,7,10,13,16,19,22,25,28,31,34,37,40,43,46,49,52,55,58,61-icosaoxo- 3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosaazatetrahexacontanoic acid, 10 KVL-11 Step A. Preparation of Int 206a To a solution of (2S,4R)-1-[(2S)-2-azido-3-methyl-butanoyl]-N-[(1S)-1-[3-fluoro-2- hydroxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]-4-hydroxy-pyrrolidine-2-carboxamide Int 7i 15 (0.33 g, 672 μmol, 1 eq) in DMF (5 mL) was added K₂CO₃ (232 mg, 1.68 mmol, 2.5 eq) and allyl N-[(1S)-2-[[(1S)-2-[4-(chloromethyl)anilino]-1-methyl-2-oxo-ethyl]amino]-1-methyl-2- oxo-ethyl]carbamate (272 mg, 740 μmol, 1.1 eq). The mixture was stirred at 20°C for 2 hrs. The reaction mixture was filtered and then diluted with H₂O (10 mL) and extracted with EtOAc (10 mL x 2). The combined organic layers were washed with brine (5 mL), dried over Na2SO4, 20 filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO₂, Petroleum ether/Ethyl acetate = 100/1 to 0/1).Compound allyl N-[(1S)-2-[[(1S)-2-[4-[[6-[(1S)-1-[[(2S,4R)-1-[(2S)-2-azido-3-methyl-butanoyl]-4-hydroxy- pyrrolidine-2-carbonyl]amino]ethyl]-2-fluoro-3-(4-methylthiazol-5-yl)phenoxy]methyl]anilino]- 1-methyl-2-oxo-ethyl]amino]-1-methyl-2-oxo-ethyl]carbamate Int 206a (0.52 g, 632 μmol, 25 94.05% yield) was obtained as a white solid.¹H NMR (400 MHz, DMSO-d6) δ10.04 (s, 1H), 9.24 - 9.08 (m, 1H), 8.64 (d, J = 7.2 Hz, 1H), 8.17 (d, J = 7.2 Hz, 1H), 7.68 (d, J = 8.4 Hz, 2H), 7.57 - 7.41 (m, 3H), 7.33 - 7.17 (m, 2H), 6.06 - 5.89 (m, 1H), 5.40 - 5.27 (m, 2H), 5.26 - 5.06 (m, 4H), 4.64 - 4.42 (m, 4H), 4.22 - 4.08 (m, 1H), 3.79 (d, J = 8.0 Hz, 1H), 3.62 - 3.51 (m, 2H), 308 15077.006WO1 2.39 (s, 3H), 2.24 - 2.08 (m, 2H), 1.79 - 1.73 (m, 1H), 1.40-1.35(m, 6H), 1.28-1.24(m, 3H), 1.07 - 1.01 (m, 6H). LCMS: MS (ESI) m/z 822.2 [M+H]⁺ Step B. Preparation of Int 206b 5 To a solution of Int 206a (0.48 g, 584 μmol, 1 eq), tert-butyl-(4-ethynoxybutoxy)- dimethyl-silane (267 mg, 1.17 mmol, 2 eq) in t-BuOH (1 mL) THF (1 mL) and H2O (1 mL) was added copper;sulfate (46.6 mg, 292 μmol, 44.8 μL, 0.5 eq) and sodium;(2R)-2-[(1S)-1,2- dihydroxyethyl]-4-hydroxy-5-oxo-2H-furan-3-olate (173 mg, 876 μmol, 1.5 eq). The mixture was stirred at 30°C for 2 hr. The reaction mixture was diluted with H2O (25 mL) and extracted 10 with EtOAc (30 mL x 3). The combined organic layers were washed with brine (50 mL), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. Compound allyl N-[(1S)-2-[[(1S)-2-[4-[[6-[(1S)-1-[[(2S,4R)-1-[(2S)-2-[4-[4-[tert- butyl(dimethyl)silyl]oxybutoxy]triazol-1-yl]-3-methyl-butanoyl]-4-hydroxy-pyrrolidine-2- carbonyl]amino]ethyl]-2-fluoro-3-(4-methylthiazol-5-yl)phenoxy]methyl]anilino]-1-methyl-2- 15 oxo-ethyl]amino]-1-methyl-2-oxo-ethyl]carbamate Int 206b (0.6 g, crude) was obtained as a yellow solid. LCMS: MS (ESI) m/z 1050.3 [M+H]⁺ Step C. Preparation of Int 206c To a solution of Int 206b (0.6 g, 571 μmol, 1 eq) in DCM (10 mL) was added Ac₂O 20 (87.5 mg, 856 μmol, 80.5 μL, 1.5 eq), DMAP (6.98 mg, 57.1 μmol, 0.1 eq) and TEA (173 mg, 1.71 mmol, 238 μL, 3 eq). The mixture was stirred at 20°C for 1 hr. The reaction mixture was 309 15077.006WO1 diluted with H2O (25 mL) and extracted with EtOAc (30 mL x 3). The combined organic layers were washed with brine (20 mL), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The crude product [(3R,5S)-5-[[(1S)-1-[2-[[4-[[(2S)-2-[[(2S)-2- (allyloxycarbonylamino)propanoyl]amino]propanoyl]amino]phenyl]methoxy]-3-fluoro-4-(4- 5 methylthiazol-5-yl)phenyl]ethyl]carbamoyl]-1-[(2S)-2-[4-[4-[tert- butyl(dimethyl)silyl]oxybutoxy]triazol-1-yl]-3-methyl-butanoyl]pyrrolidin-3-yl] acetate Int 206c (0.62 g, crude) was used into the next step without further purification. LCMS: MS (ESI) m/z 1092.7 [M+H]⁺ Step D. Preparation of Int 206d 10 To a solution of Int 206c (0.62 g, 567 μmol, 1 eq) in MeOH (7 mL) was added acetyl chloride (66.8 mg, 851 μmol, 60.5 μL, 1.5 eq). The mixture was stirred at 20°C for 1 hr. The reaction mixture was diluted with NaHCO₃ (10 mL) and extracted with DCM (20 mL x 2). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered and15 concentrated under reduced pressure to give a residue. The crude product [(3R,5S)-5-[[(1S)-1- [2-[[4-[[(2S)-2-[[(2S)-2- (allyloxycarbonylamino)propanoyl]amino]propanoyl]amino]phenyl]methoxy]-3-fluoro-4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]-1-[(2S)-2-[4-(4-hydroxybutoxy)triazol-1-yl]-3- methyl-butanoyl]pyrrolidin-3-yl] acetate Int 206d (0.55 g, 562 μmol, 99.07% yield) was used 20 into the next step without further purification. LCMS: MS (ESI) m/z 978.3 [M+H]⁺ Step E. Preparation of Int 206e 310 15077.006WO1 To a solution of Int 206d (0.55 g, 562 μmol, 1 eq) in DCM (6 mL) was added Dess- Martin (357 mg, 843 μmol, 261 μL, 1.5 eq). The mixture was stirred at 20°C for 1 hr. The reaction mixture was diluted with NaHCO3 (10 mL) and extracted with DCM (20 mL x 2). The 5 combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, Petroleum ether/Ethyl acetate=10/1 to 0/1). Compound [(3R,5S)-5- [[(1S)-1-[2-[[4-[[(2S)-2-[[(2S)-2- (allyloxycarbonylamino)propanoyl]amino]propanoyl]amino]phenyl]methoxy]-3-fluoro-4-(4-10 methylthiazol-5-yl)phenyl]ethyl]carbamoyl]-1-[(2S)-3-methyl-2-[4-(4-oxobutoxy)triazol-1- yl]butanoyl]pyrrolidin-3-yl] acetate Int 206e (0.4 g, 409 μmol, 72.88% yield) was obtained as a white solid.¹H NMR (400 MHz, DMSO-d6) δ10.00 (s, 1H), 9.71 (s, 1H), 9.12 (s, 1H), 8.61 (d, J = 7.2 Hz, 1H), 8.12 (d, J = 7.2 Hz, 1H), 8.06 - 7.95 (m, 1H), 7.85 (d, J = 8.0 Hz, 1H), 7.80 - 7.61 (m, 3H), 7.54 - 7.34 (m, 3H), 7.27 - 7.08 (m, 2H), 6.05 - 5.78 (m, 1H), 5.42 - 4.98 (m, 7H), 4.54 15 - 4.29 (m, 4H), 4.17 - 3.99 (m, 6H), 2.34 (s, 3H), 2.06 - 1.87 (m, 5H), 1.79 - 1.67 (m, 1H), 1.65 - 1.49 (m, 2H), 1.38 - 1.14 (m, 9H), 1.10 - 0.96 (m, 3H), 0.70 (d, J = 6.4 Hz, 3H). LCMS: MS (ESI) m/z 976.4 [M+H]⁺ Step F. Preparation of Int 206f 20 To a solution of Int 206e (281 mg, 288 μmol, 1.3 eq) and (4S)-2-amino-4-methyl-4-[3- [2-[(2S)-2-methyl-1,4-diazepan-1-yl]pyrimidin-4-yl]-1,2,4-oxadiazol-5-yl]-6,7-dihydro-5H- 311 15077.006WO1 benzothiophene-3-carbonitrile (100 mg, 222 μmol, 1 eq) in MeOH (6 mL) was added NaBH3CN (55.8 mg, 888 μmol, 4 eq). The mixture was stirred at 20°C for 2 hrs. The reaction mixture was diluted with H₂O (20 mL) and extracted with DCM (20 mL x 2). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered and concentrated under reduced 5 pressure to give a residue. The residue was purified by column chromatography (SiO2, Petroleum ether/Ethyl acetate = 100/1 to 0/1). Compound [(3R,5S)-5-[[(1S)-1-[2-[[4-[[(2S)-2- [[(2S)-2-(allyloxycarbonylamino)propanoyl]amino]propanoyl]amino]phenyl]methoxy]-3-fluoro- 4-(4-methylthiazol-5-yl)phenyl]ethyl]carbamoyl]-1-[(2S)-2-[4-[4-[(3S)-4-[4-[5-[(4S)-2-amino- 3-cyano-4-methyl-6,7-dihydro-5H-benzothiophen-4-yl]-1,2,4-oxadiazol-3-yl]pyrimidin-2-yl]-3- 10 methyl-1,4-diazepan-1-yl]butoxy]triazol-1-yl]-3-methyl-butanoyl]pyrrolidin-3-yl] acetate Int 206f (0.135 g, 95.7 μmol, 43.12% yield) was obtained as a white solid. LCMS: MS (ESI) m/z 1410.4 [M+H]⁺ Step G. Preparation of Int 206g 15 To a solution of Int 206f (0.13 g, 92.1 μmol, 1 eq) in MeOH (2 mL) was added K2CO3 (25.4 mg, 184 μmol, 2 eq). The mixture was stirred at 25°C for 1 hr. The reaction mixture was filtered and diluted with H2O (20 mL) and extracted with DCM (20 mL x 2). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The crude product allyl N-[(1S)-2-[[(1S)-2-[4-[[6-20 [(1S)-1-[[(2S,4R)-1-[(2S)-2-[4-[4-[(3S)-4-[4-[5-[(4S)-2-amino-3-cyano-4-methyl-6,7-dihydro- 5H-benzothiophen-4-yl]-1,2,4-oxadiazol-3-yl]pyrimidin-2-yl]-3-methyl-1,4-diazepan-1- yl]butoxy]triazol-1-yl]-3-methyl-butanoyl]-4-hydroxy-pyrrolidine-2-carbonyl]amino]ethyl]-2- fluoro-3-(4-methylthiazol-5-yl)phenoxy]methyl]anilino]-1-methyl-2-oxo-ethyl]amino]-1- methyl-2-oxo-ethyl]carbamate Int 206g (0.12 g, 87.7 μmol, 95.14% yield) was used into the 25 next step without further purification. LCMS: MS (ESI) m/z 1368.4 [M+H]⁺ Step H. Preparation of Int 206h 312 15077.006WO1 To a solution of Int 206g (0.1 g, 73.1 μmol, 1 eq) in DCM (3 mL) was added Pd(PPh₃)₄ (25.3 mg, 21.9 μmol, 0.3 eq) and DABCO (40.9 mg, 365 μmol, 40.2 μL, 5 eq). The mixture was stirred at 20°C for 5 min. The reaction solution was adjusted to pH = 6 with TFA. The residue 5 was purified by prep-HPLC (column: Phenomenex Luna C1875 x 30mm x 3um; mobile phase: [H2O(0.1% TFA)-ACN];gradient:15%-45% B over 8.0 min). Compound (2S,4R)-1-[(2S)-2-[4- [4-[(3S)-4-[4-[5-[(4S)-2-amino-3-cyano-4-methyl-6,7-dihydro-5H-benzothiophen-4-yl]-1,2,4- oxadiazol-3-yl]pyrimidin-2-yl]-3-methyl-1,4-diazepan-1-yl]butoxy]triazol-1-yl]-3-methyl- butanoyl]-N-[(1S)-1-[2-[[4-[[(2S)-2-[[(2S)-2-10 aminopropanoyl]amino]propanoyl]amino]phenyl]methoxy]-3-fluoro-4-(4-methylthiazol-5- yl)phenyl]ethyl]-4-hydroxy-pyrrolidine-2-carboxamide Int 206h (60 mg, 46.7 μmol, 63.93% yield) was obtained as a white solid.¹H NMR (400 MHz, DMSO-d₆) δ10.21 (s, 1H), 9.45 - 9.32 (m, 1H), 9.13 (s, 1H), 8.74 - 8.63 (m, 2H), 8.58 (d, J = 7.6 Hz, 1H), 8.06 (d, J = 3.6 Hz, 3H), 7.71 (s, 1H), 7.63 (d, J = 8.4 Hz, 2H), 7.47 (d, J = 8.4 Hz, 2H), 7.34 - 7.19 (m, 3H), 7.13 -7.09 15 (m, 2H), 5.28 (t, J = 7.2 Hz, 1H), 5.17 - 5.03 (m, 3H), 4.96 - 4.77 (m, 1H), 4.53 - 4.48(m, 1H), 4.46 - 4.25 (m, 3H), 4.18 - 4.08 (m, 2H), 3.94 - 3.60 (m, 6H), 2.33 (s, 3H), 2.15 - 1.61 (m, 16H), 1.49 - 1.24 (m, 9H), 1.17 - 1.12 (m, 3H), 1.06 (d, J = 6.4 Hz, 3H), 0.68 (d, J = 6.4 Hz, 3H). LCMS: MS (ESI) m/z 1284.6[M+H]⁺ 313 15077.006WO1 Step I. Preparation of KVL-11 To a solution of Int 206h (30 mg, 21.4 μmol, 1 eq, TFA) in DMF (1 mL) was added DIEA (13.8 mg, 107 μmol, 18.7 μL, 5 eq) and 64-(6-(2-carboxyethyl)-3-(2,5-dioxo-2,5-dihydro- 5 1H-pyrrol-1-yl)-2-fluorophenoxy)-3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60- icosamethyl-4,7,10,13,16,19,22,25,28,31,34,37,40,43,46,49,52,55,58,61-icosaoxo- 3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosaazatetrahexacontanoic acid Int L (39.4 mg, 20.4 μmol, 0.95 eq). The mixture was stirred at 20°C for 1 hr. The reaction solution was adjusted to pH = 6 with TFA. The residue was purified by prep-HPLC (column:10 Phenomenex Luna C1875x30mmx3um;mobile phase: [H₂O(0.1% TFA)-ACN];gradient:20%- 50% B over 8.0 min) to give KVL-11 (30 mg, 8.35 μmol, 38.93% yield) as a white solid.1H NMR (400 MHz, DMSO-d6) δ9.96 (s, 1H), 9.42-9.25 (m, 1H), 9.12 (s, 1H), 8.74-8.62 (m, 1H), 8.56 (d, J = 7.2 Hz, 1H), 8.22-8.07 (m, 2H), 7.73 (s, 1H), 7.65 (d, J = 8.0 Hz, 2H), 7.46 (d, J = 8.4 Hz, 2H), 7.33-6.97 (m, 8H), 5.31-5.20 (m, 1H), 5.16-4.99 (m, 3H), 4.98-4.79 (m, 1H), 4.47- 15 3.64 (m, 52H), 3.06-2.65 (m, 64H), 2.46 (s, 3H), 2.15-1.82 (m, 11H), 1.79 (s, 3H), 1.74 (s, 2H), 314 15077.006WO1 1.34-1.25 (m, 6H), 1.20 (d, J = 6.9 Hz, 3H), 1.05 (d, J = 6.4 Hz, 3H), 0.68 (d, J = 6.4 Hz, 3H). LCMS: MS (ESI) m/z 1077.5 [M/2+H]- Example BXVL-20 Synthesis of 64-(6-(3-(((S)-1-(((S)-1-((4-((2-((S)-1-((2S,4R)-1- ((S)-2-(4-(N-(6-(8-(benzo[d]thiazol-2-ylcarbamoyl)-3,4-dihydroisoquinolin-2(1H)-yl)-3-(1- 5 (((1r,3s,5R,7S)-3-(2-hydroxyethoxy)-5,7-dimethyladamantan-1-yl)methyl)-5-methyl-1H- pyrazol-4-yl)picolinoyl)sulfamoyl)butanamido)-3,3-dimethylbutanoyl)-4-hydroxypyrrolidine-2- carboxamido)ethyl)-5-(4-methylthiazol-5-yl)phenoxy)methyl)phenyl)amino)-1-oxopropan-2- yl)amino)-1-oxopropan-2-yl)amino)-3-oxopropyl)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2- fluorophenoxy)-3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosamethyl-10 4,7,10,13,16,19,22,25,28,31,34,37,40,43,46,49,52,55,58,61-icosaoxo- 3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosaazatetrahexacontanoic acid, BXVL-20 Step A. Preparation of Int 207a 15 To a solution of 4-[[6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydro-1H-isoquinolin- 2-yl]-3-[1-[[3-[2-[tert-butyl(dimethyl)silyl]oxyethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5- methyl-pyrazol-4-yl]pyridine-2-carbonyl]sulfamoyl]butanoic acid, Int.8f (500 mg, 495 μmol, 1 eq) in DCM (5 mL) was added 2,3,5,6-tetrafluorophenol (329 mg, 1.98 mmol, 4 eq) and EDCI (380 mg, 1.98 mmol, 4 eq). The mixture was stirred at 20 °C for 30min. The reaction mixture 20 was quenched by addition H2O 10 mL at 20 °C, and extracted with DCM (15 mL * 3). The combined organic layers were washed with brine (15 mL), dried over [Na2SO4], filtered and concentrated under reduced pressure to give compound (2,3,5,6-tetrafluorophenyl) 4-[[6-[8-(1,3- benzothiazol-2-ylcarbamoyl)-3,4-dihydro-1H-isoquinolin-2-yl]-3-[1-[[3-[2-[tert- butyl(dimethyl)silyl]oxyethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4- 25 yl]pyridine-2-carbonyl]sulfamoyl]butanoate Int 207a (900 mg, crude) was obtained as yellow solid. LCMS: MS (ESI) m/z 1158.2 [M+H]⁺ Step B. Preparation of Int 207b 315 15077.006WO1 To a solution of Int 207a (40.0 mg, 44.1 μmol, 1 eq, TFA) in DMF (0.50 mL) was added DIEA (17.0 mg, 132 μmol, 23.0 μL, 3 eq) and (2,3,5,6-tetrafluorophenyl) 4-[[6-[8-(1,3- benzothiazol-2-ylcarbamoyl)-3,4-dihydro-1H-isoquinolin-2-yl]-3-[1-[[3-[2-[tert- 5 butyl(dimethyl)silyl]oxyethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4- yl]pyridine-2-carbonyl]sulfamoyl]butanoate (81.0 mg, 44.1 μmol, 1 eq). The mixture was stirred at 20°C for 1 hr. The reaction mixture was quenched by addition H₂O 5 mL at 20°C, and extracted with Ethyl acetate (5 mL x 3). The combined organic layers were washed with brine (10 mL x 3), dried over Na2SO4, filtered and concentrated under reduced pressure to give allyl10 N-[(1S)-2-[[(1S)-2-[4-[[2-[(1S)-1-[[(2S,4R)-1-[(2S)-2-[4-[[6-[8-(1,3-benzothiazol-2- ylcarbamoyl)-3,4-dihydro-1H-isoquinolin-2-yl]-3-[1-[[3-[2-[tert- butyl(dimethyl)silyl]oxyethoxy]-5,7-dimethyl-1-adamantyl]methyl]-5-methyl-pyrazol-4- yl]pyridine-2-carbonyl]sulfamoyl]butanoylamino]-3,3-dimethyl-butanoyl]-4-hydroxy- pyrrolidine-2-carbonyl]amino]ethyl]-5-(4-methylthiazol-5-yl)phenoxy]methyl]anilino]-1- 15 methyl-2-oxo-ethyl]amino]-1-methyl-2-oxo-ethyl]carbamate Int 207b (290 mg, crude) as yellow oil. LCMS: MS (ESI) m/z 892.9 [1/2M+H]⁺ Step C. Preparation of Int 207c 316 15077.006WO1 To a solution of Int 207b (270 mg, 151 μmol, 1 eq) in MeOH (2.00 mL) was added acetyl chloride (36.0 mg, 453 μmol, 32.2 μL, 3 eq). The mixture was stirred at 20°C for 1 hr. The reaction mixture was quenched by addition H2O 5 mL at 20°C, and extracted with Ethyl 5 acetate (10 mL x 3). The combined organic layers were washed with brine (10 mL x 1), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give allyl N-[(1S)-2-[[(1S)-2- [4-[[2-[(1S)-1-[[(2S,4R)-1-[(2S)-2-[4-[[6-[8-(1,3-benzothiazol-2-ylcarbamoyl)-3,4-dihydro-1H- isoquinolin-2-yl]-3-[1-[[3-(2-hydroxyethoxy)-5,7-dimethyl-1-adamantyl]methyl]-5-methyl- pyrazol-4-yl]pyridine-2-carbonyl]sulfamoyl]butanoylamino]-3,3-dimethyl-butanoyl]-4-hydroxy-10 pyrrolidine-2-carbonyl]amino]ethyl]-5-(4-methylthiazol-5-yl)phenoxy]methyl]anilino]-1- methyl-2-oxo-ethyl]amino]-1-methyl-2-oxo-ethyl]carbamate Int 207c (77.0 mg, 46.1 μmol, 30.4% yield) as white solid. LCMS: MS (ESI) m/z 835.8 [1/2M+H]⁺ Step D. Preparation of Int 207d 317 15077.006WO1 To a solution of Int 207c (74.8 mg, 44.8 μmol, 1 eq) in DCM (1.00 mL) was added 1,4- diazabicyclo[2.2.2]octane (25.0 mg, 224 μmol, 24.6 μL, 5 eq) and palladium;triphenylphosphane (1.65 mg, 4.48 μmol, 0.10 eq). The mixture was stirred at 20°C 5 for 1 hr. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC ([H₂O(0.1% TFA)-ACN];gradient:25%-60% B over 8.0 min ) to give 2-[6-[[4-[[(1S)-1-[(2S,4R)-2-[[(1S)-1-[2-[[4-[[(2S)-2-[[(2S)-2- aminopropanoyl]amino]propanoyl]amino]phenyl]methoxy]-4-(4-methylthiazol-5- yl)phenyl]ethyl]carbamoyl]-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl-propyl]amino]-4-10 oxo-butyl]sulfonylcarbamoyl]-5-[1-[[3-(2-hydroxyethoxy)-5,7-dimethyl-1-adamantyl]methyl]- 5-methyl-pyrazol-4-yl]-2-pyridyl]-N-(1,3-benzothiazol-2-yl)-3,4-dihydro-1H-isoquinoline-8- carboxamide Int 207d (12 mg, 7.57 μmol, 16.8% yield) as white solid. LCMS: MS (ESI) m/z 793.7 [1/2M+H]⁺ Step E. Preparation of BXVL-20 318 15077.006WO1 To a solution of Int 207d (12.0 mg, 7.57 μmol, 1 eq) in DMF (1.00 mL) was added DIEA (2.90 mg, 22.7 μmol, 3.95 μL, 3 eq) and 64-(6-(2-carboxyethyl)-3-(2,5-dioxo-2,5- dihydro-1H-pyrrol-1-yl)-2-fluorophenoxy)- 5 3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosamethyl- 4,7,10,13,16,19,22,25,28,31,34,37,40,43,46,49,52,55,58,61-icosaoxo- 3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosaazatetrahexacontanoic acid Int L (16.0 mg, 8.32 μmol, 1.10 eq). The mixture was stirred at 20°C for 1 hr. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by 10 prep-HPLC ([H₂O(0.1% TFA)-ACN];gradient:25%-60% B over 8.0 min ) to give BXVL-20 (6.50 mg, 1.81 μmol, 23.8% yield, 93.2% purity) as white solid.1H NMR (400 MHz, DMSO- d6) δ12.87 - 12.84(m, 1H), 11.74 - 11.68 (m, 1H), 9.98 - 9.90 (m, 1H), 8.97 (s, 1H), 8.35 - 8.26 (m, 1H), 8.17 - 8.07 (m, 2H), 8.02 (d, J = 8.0 Hz, 1H), 7.94 - 7.87 (m, 1H), 7.83 - 7.75 (m, 1H), 7.63 (d, J = 8.0 Hz, 3H), 7.54 (d, J = 8.8 Hz, 1H), 7.49 - 7.27 (m, 9H), 7.24 (s, 2H), 7.08 (s, 2H), 15 7.05 - 6.97 (m, 3H), 5.23 - 5.19 (m, 1H), 5.16 (s, 2H), 4.95 (s, 2H), 4.54 - 4.43 (m, 4H), 4.36 - 4.18 (m, 24H), 4.13 - 3.89 (m, 28H), 2.96 - 2.73 (m, 68H), 2.40 (s, 3H), 2.09 (s, 3H), 1.99 - 1.78 (m, 6H), 1.37 - 1.13 (m, 21H), 0.93 (s, 9H), 0.84 (s, 6H). LCMS: MS (ESI) m/z 1678.2 [1/2M+H]⁺ 319 15077.006WO1 The following compounds were prepared in a manner similar to that described for BXVL-20 using the appropriate compound as starting material. 320 15077.006WO1 Example WVL-1 Synthesis of 64-(6-(3-(((S)-1-(((S)-1-((4-((6-((S)-1-((2S,4R)-1- ((S)-2-(2-((6-(4-(4-((2-allyl-1-(6-(2-hydroxypropan-2-yl)pyridin-2-yl)-3-oxo-2,3-dihydro-1H- pyrazolo[3,4-d]pyrimidin-6-yl)amino)phenyl)piperazin-1-yl)hexyl)oxy)acetamido)-3,3- 5 dimethylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)ethyl)-2-fluoro-3-(4-methylthiazol-5- yl)phenoxy)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-1-oxopropan-2-yl)amino)-3- oxopropyl)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-fluorophenoxy)- 3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosamethyl- 4,7,10,13,16,19,22,25,28,31,34,37,40,43,46,49,52,55,58,61-icosaoxo- 10 3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosaazatetrahexacontanoic acid, WVL-1 Step A. Preparation of Int 208a 321 15077.006WO1 To a solution of tert-butyl N-[(1S)-1-[(2S,4R)-2-[[(1S)-1-[3-fluoro-2-hydroxy-4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl]carbamate Int 7g (0.2 g, 346 μmol, 1 eq) and allyl N-[(1S)-2-[[(1S)-2-[4- 5 (chloromethyl)anilino]-1-methyl-2-oxo-ethyl]amino]-1-methyl-2-oxo-ethyl]carbamate (318 mg, 864 μmol, 2.5 eq) in DMF (3 mL) was added K2CO3 (143 mg, 1.04 mmol, 3 eq). The reaction mixture was stirred at 20°C for 2 hr. The residue was diluted with water 20 mL and extracted with EtOAc 6 ml x3. The combined organic layers were washed with brine 20 mL, dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was 10 purified by column chromatography (SiO₂, Petroleum ether/Ethyl acetate=1/0 to 0/1). Compound tert-butyl N-[(1S)-1-[(2S,4R)-2-[[(1S)-1-[2-[[4-[[(2S)-2-[[(2S)-2- (allyloxycarbonylamino) propanoyl] amino] propanoyl] amino] phenyl] methoxy]-3-fluoro-4-(4- methylthiazol-5-yl) phenyl] ethyl] carbamoyl]-4-hydroxy-pyrrolidine-1-carbonyl]-2,2-dimethyl- propyl] carbamate Int 208a (180 mg, 198 μmol, 57.23% yield) was obtained as a brown solid. 15 LCMS: MS (ESI) m/z 910.4 [M+H]+ Step B. Preparation of Int 208b A solution of Int 208a (0.16 g, 176 μ mol, 1 eq) in HCOOH (8.45 mg, 176 μ mol, 1 eq) was stirred at 20°C for 2 hr. The reaction mixture was concentrated under reduced pressure to 20 remove HCOOH. The residue was purified by prep-HPLC (column: Phenomenex Luna C18 75*30mm*3 um; mobile phase: [H₂O (0.1%TFA)-ACN]; gradient:20%-50% B over 8.0 min). 322 15077.006WO1 Compound allyl N-[(1S)-2-[[(1S)-2-[4-[[6-[(1S)-1-[[(2S,4R)-1-[(2S)-2-amino-3,3-dimethyl- butanoyl]-4-hydroxy-pyrrolidine-2-carbonyl] amino] ethyl]-2-fluoro-3-(4-methylthiazol-5-yl) phenoxy] methyl] anilino]-1-methyl-2-oxo-ethyl] amino]-1-methyl-2-oxo-ethyl] carbamate Int 208b (0.08 g, 98.8 μ mol, 28.09% yield) was obtained as a white solid. LCMS: MS (ESI) m/z 5 810.5 [M+H]+ Step C. Preparation of Int 208c To a solution of Int 208b (75.0 mg, 92.6 μ mol, 1 eq) in DMF (1 mL) were added HATU (52.8 mg, 139 μmol, 1.5 eq), DIEA (60.0 mg, 463 μmol, 80.6 μL, 5 eq) and 2-[6-[4-[4-[[2-allyl-10 1-[6-(1-hydroxy-1-methyl-ethyl)-2-pyridyl]-3-oxo-pyrazolo[3,4-d]pyrimidin-6- yl]amino]phenyl]piperazin-1-yl]hexoxy]acetic acid Int 9e (65.7 mg, 102 μmol, 1.1 eq). The reaction mixture was stirred at 20°C for 1 hr. The reaction mixture was quenched by addition water 3 mL and extracted with EtOAc 5 mL x 2. The combined organic layers were dried over brine, filtered and concentrated under reduced pressure to give a residue. Compound allyl N-15 [(1S)-2-[[(1S)-2-[4-[[6-[(1S)-1-[[(2S,4R)-1-[(2S)-2-[[2-[6-[4-[4-[[2-allyl-1-[6-(1-hydroxy-1- methyl-ethyl)-2-pyridyl]-3-oxo-pyrazolo[3,4-d]pyrimidin-6-yl]amino] phenyl] piperazin-1-yl] hexoxy] acetyl] amino]-3,3-dimethyl-butanoyl]-4-hydroxy-pyrrolidine-2-carbonyl] amino] 323 15077.006WO1 ethyl]-2-fluoro-3-(4-methylthiazol-5-yl) phenoxy] methyl] anilino]-1-methyl-2-oxo-ethyl] amino]-1-methyl-2-oxo-ethyl]carbamate Int 208c (0.10 g, crude) was obtained as a white solid and used into the next step without further purification. LCMS: MS (ESI) m/z 1436.6 [M+H]+ Step D. Preparation of Int 208d 5 To a solution of Int 208c (0.08 g, 55.7 μmol, 1 eq) in DCM (2 mL) was added Pd(PPh₃)₄ (19.3 mg, 16.7 μmol, 0.3 eq) and DABCO (50 mg, 445 μmol, 49 μL, 8 eq). The reaction mixture was stirred at 20°C for 1hr. The mixture was adjusted pH to 5 with TFA, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC10 (column: Phenomenex luna C18100*40mm*3 um;mobile phase: [H₂O(0.1%TFA)- ACN];gradient:15%-45% B over 8.0 min). Compound (2S,4R)-1-[(2S)-2-[[2-[6-[4-[4-[[2-allyl- 1-[6-(1-hydroxy-1-methyl-ethyl)-2-pyridyl]-3-oxo-pyrazolo [3,4-d] pyrimidin-6-yl] amino] phenyl] piperazin-1-yl] hexoxy] acetyl] amino]-3,3-dimethyl-butanoyl]-N-[(1S)-1-[2-[[4-[[(2S)- 2-[[(2S)-2-aminopropanoyl] amino] propanoyl] amino] phenyl] methoxy]-3-fluoro-4-(4- 15 methylthiazol-5-yl)phenyl] ethyl]-4-hydroxy-pyrrolidine-2-carboxamide Int 208d (54.0 mg, 40.0 μmol, 71.70% yield) was obtained as a white solid.¹ H NMR (400 MHz, DMSO-d6) δ10.19 (s, 1H), 9.12 (s, 1H), 8.85 (s, 1H), 8.70 (d, J = 7.2 Hz, 1H), 8.51 (d, J = 7.6 Hz, 1H), 8.09-7.98 (m, 3H), 7.75 (d, J = 7.6 Hz, 1H), 7.65-7.57 (m, 4H), 7.45 (d, J = 8.4 Hz, 2H), 7.36 (d, J = 9.6 Hz, 1H), 7.27-7.15 (m, 2H), 7.00 (d, J = 8.8 Hz, 2H), 5.72-5.60 (m, 1H), 5.30-5.22 (m, 324 15077.006WO1 1H), 5.12-5.02 (m, 2H), 4.99 (d, J = 9.6 Hz, 1H), 4.82 (d, J = 17.2 Hz, 1H), 4.68 (d, J = 4.8 Hz, 2H), 4.56 (d, J = 9.6 Hz, 1H), 4.53-4.47 (m, 1H), 4.43-4.37 (m, 1H), 4.27 (s, 2H), 3.94 (s, 4H), 3.79 (d, J = 12.0 Hz, 4H), 3.60 (d, J = 13.2 Hz, 4H), 3.49 (t, J = 6.4 Hz, 2H), 3.47-3.40 (m, 1H), 3.15 (d, J = 6.4 Hz, 4H), 2.99-2.89 (m, 2H), 2.32 (s, 3H), 2.09-1.99 (m, 1H), 1.78-1.64 (m, 3H), 5 1.64-1.53 (m, 2H), 1.46 (s, 6H), 1.36-1.32 (m, 10H), 0.96 (s, 9H) LCMS: MS (ESI) m/z 1353.0 [M+H]+ Step E. Preparation of WVL-1 To a solution of Int 208d (12.1 mg, 8.27 μmol, 1 eq, TFA) and 64-(3-(2,5-dioxo-2,5-10 dihydro-1H-pyrrol-1-yl)-2-fluoro-6-(3-oxo-3-(2,3,5,6-tetrafluorophenoxy)propyl)phenoxy)- 3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosamethyl- 4,7,10,13,16,19,22,25,28,31,34,37,40,43,46,49,52,55,58,61-icosaoxo- 3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosaazatetrahexacontanoic acid Int L (16.0 mg, 8.27 μmol, 1 eq) in DMF (1 mL) was added DIEA (5.34 mg, 41.4 μmol, 7.20 μL, 5 15 eq). The reaction mixture was stirred at 20°C for 1 hr. The mixture was adjusted pH to 5 with TFA, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: Phenomenex luna C18100*40mm*3 um;mobile phase: 325 15077.006WO1 [H2O(0.1%TFA)-ACN];gradient:20%-50% B over 8.0 min) to give WVL-1 (10.0 mg, 3.09 μmol, 37.38% yield, TFA) as a white solid.¹H NMR (400 MHz, DMSO-d₆) δ10.15-10.14 (m, 1H), 9.96 (d, J = 2.8 Hz, 1H), 9.43-9.32 (m, 1H), 9.11 (s, 1H), 8.84 (s, 1H), 8.52-8.46 (m, 1H), 8.17-8.09 (m, 2H), 8.02 (t, J = 8.0 Hz, 1H), 7.75 (d, J = 8.4 Hz, 1H), 7.62 (t, J = 7.6 Hz, 4H), 5 7.44 (d, J = 8.4 Hz, 2H), 7.33 (d, J = 9.2 Hz, 1H), 7.24 (s, 2H), 7.22-7.16 (m, 2H), 7.15-7.07 (m, 1H), 7.06-6.96 (m, 3H), 5.72-5.59 (m, 1H), 5.28-5.21 (m, 1H), 5.06 (d, J = 3.6 Hz, 2H), 4.99 (d, J = 10.0 Hz, 1H), 4.82 (d, J = 16.8 Hz, 1H), 4.72-4.65 (m, 2H), 4.55 (d, J = 10.0Hz, 1H), 4.46- 4.17 (m, 22H), 4.14-3.88 (m, 23H), 4.48-3.84 (m, 1H), 3.78 (dd, J = 1.2, 13.2 Hz, 3H), 3.62- 3.55 (m, 6H), 3.19-3.10 (m, 5H), 3.20-3.10 (m, 1H), 3.00-2.64 (m, 64H), 2.34-2.30 (m, 5H),10 2.10-1.98 (m, 2H), 1.96-1.89 (m, 2H), 1.75-1.65 (m, 3H), 1.63-1.55 (m, 2H), 1.46 (s, 6H), 1.39- 1.36 (m, 4H), 1.33-1.26 (m, 6H), 1.23-1.18 (m, 3H), 0.95 (s, 9H). LCMS: MS (ESI) m/z 3118.49 [M-H]- The following compounds were prepared in a manner similar to that described for WVL- 1 using the appropriate compound as starting material. 326 15077.006WO1 Example WVL-6 Synthesis of 64-(6-(3-(((S)-1-(((S)-1-((4-((2-((S)-1-((2S,4R)-1- ((S)-2-(4-((6-(4-(4-((2-allyl-1-(6-(2-hydroxypropan-2-yl)pyridin-2-yl)-3-oxo-2,3-dihydro-1H- pyrazolo[3,4-d]pyrimidin-6-yl)amino)phenyl)piperazin-1-yl)hexyl)oxy)-1H-1,2,3-triazol-1-yl)- 5 3-methylbutanoyl)-4-hydroxypyrrolidine-2-carboxamido)ethyl)-5-(4-methylthiazol-5- yl)phenoxy)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-1-oxopropan-2-yl)amino)-3- oxopropyl)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-fluorophenoxy)- 3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosamethyl- 4,7,10,13,16,19,22,25,28,31,34,37,40,43,46,49,52,55,58,61-icosaoxo- 10 3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosaazatetrahexacontanoic acid, WVL-6 Step A. Preparation of Int 209a To a solution of (2S,4R)-1-[(2S)-2-azido-3-methyl-butanoyl]-4-hydroxy-N-[(1S)-1-[2- 15 hydroxy-4-(4-methylthiazol-5-yl)phenyl]ethyl]pyrrolidine-2-carboxamide Int 7x (400 mg, 846.45 μmol, 1 eq) in DMF (4 mL) were added allyl N-[(1S)-2-[[(1S)-2-[4- (chloromethyl)anilino]-1-methyl-2-oxo-ethyl]amino]-1-methyl-2-oxo-ethyl]carbamate (467 mg, 1.27 mmol, 1.5 eq) and K₂CO₃ (350 mg, 2.54 mmol, 3 eq). The reaction mixture was stirred at 25°C for 1 hr. The reaction mixture was added H₂O 30 mL and extracted with ethyl acetate 20 (30 mL x 3), the organic layer was washed with brine (30 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO₂, Ethyl acetate/MeOH =100/0 to 50/50) to give allyl N-[(1S)-2-[[(1S)-2- [4-[[2-[(1S)-1-[[(2S,4R)-1-[(2S)-2-azido-3-methyl-butanoyl]-4-hydroxy-pyrrolidine-2- carbonyl]amino]ethyl]-5-(4-methylthiazol-5-yl)phenoxy]methyl]anilino]-1-methyl-2-oxo- 25 ethyl]amino]-1-methyl-2-oxo-ethyl]carbamate Int 209a (0.38 g, 472 μmol, 55.8% yield) as a brown solid. LCMS: MS (ESI) m/z 804.2[M+H]⁺ Step B. Preparation of Int 209b 327 15077.006WO1 To a solution of Int 209a (0.36 g, 447 μmol, 1 eq) in THF:t-BuOH:H₂O=1:1:1 (4 mL) were added tert-butyl-(6-ethynoxyhexoxy)-dimethyl-silane (172 mg, 671 μmol, 1.5 eq), sodium;(2R)-2-[(1S)-1,2-dihydroxyethyl]-4-hydroxy-5-oxo-2H-furan-3-olate (133 mg, 671 5 μmol, 1.5 eq) and CuSO4 (35.7 mg, 223 μmol, 34.3 μL, 0.5 eq). The reaction mixture was heated to 30^oC and stirred for 2 hr. After cooling to room temperature, the mixture was water (10 mL) was added and then extracted with ethyl acetate (30 mL x 3). The combined organic phases were washed with brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to dryness. The residue was purified by column 10 chromatography (SiO₂, Ethyl acetate/MeOH=100/0 to 50/50) to give allyl N-[(1S)-2-[[(1S)-2-[4- [[2-[(1S)-1-[[(2S,4R)-1-[(2S)-2-[4-[6-[tert-butyl(dimethyl)silyl]oxyhexoxy]triazol-1-yl]-3- methyl-butanoyl]-4-hydroxy-pyrrolidine-2-carbonyl]amino]ethyl]-5-(4-methylthiazol-5- yl)phenoxy]methyl]anilino]-1-methyl-2-oxo-ethyl]amino]-1-methyl-2-oxo-ethyl]carbamate Int 209b (0.26 g, 245.20 μmol, 54.76% yield) as a brown solid. LCMS: MS (ESI) m/z 15 1060.4[M+H]⁺ Step C. Preparation of Int 209c To a solution of Int 209b (240 mg, 226 μmol, 1 eq) in DCM (3 mL) were added Ac₂O (57.7 mg, 565 μmol, 53.1 μL, 2.5 eq), TEA (91.6 mg, 905 μmol, 126 μL, 4 eq) and DMAP (2.7 20 mg, 22.6 μmol, 0.1 eq). The reaction mixture was stirred at 25°C for 30 min. The reaction mixture was added H₂O 20 mL and extracted with DCM (20 mL x 3), the organic layer was washed with brine (10 mL), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give [(3R,5S)-5-[[(1S)-1-[2-[[4-[[(2S)-2-[[(2S)-2- (allyloxycarbonylamino)propanoyl]amino]propanoyl]amino]phenyl]methoxy]-4-(4-25 methylthiazol-5-yl)phenyl]ethyl]carbamoyl]-1-[(2S)-2-[4-[6-[tert- 328 15077.006WO1 butyl(dimethyl)silyl]oxyhexoxy]triazol-1-yl]-3-methyl-butanoyl]pyrrolidin-3-yl] acetate Int 209c (310 mg, crude) as a brown solid. LCMS: MS (ESI) m/z 1102.4[M+H]⁺ Step D. Preparation of Int 209d 5 To a solution of Int 209c (310 mg, 281 μmol, 1 eq) in MeOH (3 mL) was added dropwise acetyl chloride (33.1 mg, 421 μmol, 29.9 μL, 1.5 eq). The reaction mixture was stirred at 25°C for 30 min. The reaction mixture was added H2O 20 mL and extracted with ethyl acetate (20 mL x 3), the organic layer was washed with brine (10 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give [(3R,5S)-5-[[(1S)-1-[2-[[4-[[(2S)-2-10 [[(2S)-2-(allyloxycarbonylamino)propanoyl]amino]propanoyl]amino]phenyl]methoxy]-4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]-1-[(2S)-2-[4-(6-hydroxyhexoxy)triazol-1-yl]-3- methyl-butanoyl]pyrrolidin-3-yl] acetate Int 209d (0.27 g, crude) as a brown solid. LCMS: MS (ESI) m/z 988.3[M+H]⁺ Step E. Preparation of Int 209e 15 To a solution of Int 209d (250 mg, 253 μmol, 1 eq) in DCM (3 mL) was added Dess- Martin (322 mg, 759 μmol, 235 μL, 3 eq). The mixture was stirred at 25°C for 1 hr. The reaction mixture was added NaHCO320 mL and extracted with DCM (20 mL x 3), the organic layer was washed with brine (10 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (SiO₂, Petroleum20 ether/Ethyl acetate = 0:1) to give [(3R,5S)-5-[[(1S)-1-[2-[[4-[[(2S)-2-[[(2S)-2- (allyloxycarbonylamino)propanoyl]amino]propanoyl]amino]phenyl]methoxy]-4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]-1-[(2S)-3-methyl-2-[4-(6-oxohexoxy)triazol-1- yl]butanoyl]pyrrolidin-3-yl] acetate Int 209e (0.115 g, 116 μmol, 46.1% yield) as a yellow solid. LCMS: MS (ESI) m/z 986.3[M+H]⁺ 25 Step F. Preparation of Int 209f To a solution of 2-allyl-1-[6-(1-hydroxy-1-methyl-ethyl)-2-pyridyl]-6-(4-piperazin-1- ylanilino)pyrazolo[3,4-d]pyrimidin-3-one (45 mg, 86.0 μmol, 1 eq, HCl) in DCM (1 mL) was added TEA (26.1 mg, 258 μmol, 35.9 μL, 3 eq), then AcOH (10.3 mg, 172 μmol, 9.85 μL, 2 eq) was added to adjust pH to 5~6, then Int 209e (93.3 mg, 94.6 μmol, 1.1 eq) and 30 NaBH(OAc)3 (27.3 mg, 129 μmol, 1.5 eq) was added. The reaction mixture was stirred 329 15077.006WO1 at 25°C for 1 hr. The reaction mixture was added H2O 10 mL and extracted with DCM (10 mL x 3), the organic layer was washed with brine (10 mL), dried over Na₂SO₄, filtered and concentrated to dryness. The residue was purified by prep-TLC (SiO_2, Ethyl acetate: MeOH = 4:1) to give [(3R,5S)-1-[(2S)-2-[4-[6-[4-[4-[[2-allyl-1-[6-(1-hydroxy-1-methyl-ethyl)-2- 5 pyridyl]-3-oxo-pyrazolo[3,4-d]pyrimidin-6-yl]amino]phenyl]piperazin-1-yl]hexoxy]triazol-1- yl]-3-methyl-butanoyl]-5-[[(1S)-1-[2-[[4-[[(2S)-2-[[(2S)-2- (allyloxycarbonylamino)propanoyl]amino]propanoyl]amino]phenyl]methoxy]-4-(4- methylthiazol-5-yl)phenyl]ethyl]carbamoyl]pyrrolidin-3-yl] acetate Int 209f (75 mg, 51.5 μmol, 59.8% yield) as a yellow solid. LCMS: MS (ESI) m/z 1456.3[M+H]⁺ 10 Step G. Preparation of Int 209g 330 15077.006WO1 To a solution of Int 209f (65 mg, 44.6 μmol, 1 eq) in MeOH (1 mL) was added K2CO3 (18.5 mg, 133 μmol, 3 eq). The reaction mixture was stirred at 25°C for 1 hr. The reaction mixture was filtered off over celite and the filter cake was washed with MeOH (50 mL x 2), then 5 concentrated under reduced pressure to give allyl N-[(1S)-2-[[(1S)-2-[4-[[2-[(1S)-1-[[(2S,4R)-1- [(2S)-2-[4-[6-[4-[4-[[2-allyl-1-[6-(1-hydroxy-1-methyl-ethyl)-2-pyridyl]-3-oxo-pyrazolo[3,4- d]pyrimidin-6-yl]amino]phenyl]piperazin-1-yl]hexoxy]triazol-1-yl]-3-methyl-butanoyl]-4- hydroxy-pyrrolidine-2-carbonyl]amino]ethyl]-5-(4-methylthiazol-5-yl)phenoxy]methyl]anilino]- 1-methyl-2-oxo-ethyl]amino]-1-methyl-2-oxo-ethyl]carbamate Int 209g (0.065 g, crude) as a 10 yellow solid. LCMS: MS (ESI) m/z 1414.3[M+H]⁺ Step H. Preparation of Int 209h 331 15077.006WO1 To a solution of Int 209g (55 mg, 38.8 μmol, 1 eq) in DCM (0.5 mL) were added DABCO (21.8 mg, 194 μmol, 21.3 μL, 5 eq) and Pd(PPh3)4 (9.0 mg, 7.78 μmol, 0.2 eq). The reaction mixture was stirred at 25°C for 5 min. The mixture was adjust pH to 5~6 with TFA 5 and filtered, then the filtrate was purified by pre-HPLC (TFA condition; column: Phenomenex Luna C1875*30mm*3um;mobile phase: [H₂O(0.1% TFA)-ACN];gradient:15%-50% B over 8.0 min ) to give (2S,4R)-1-[(2S)-2-[4-[6-[4-[4-[[2-allyl-1-[6-(1-hydroxy-1-methyl-ethyl)-2- pyridyl]-3-oxo-pyrazolo[3,4-d]pyrimidin-6-yl]amino]phenyl]piperazin-1-yl]hexoxy]triazol-1- yl]-3-methyl-butanoyl]-N-[(1S)-1-[2-[[4-[[(2S)-2-[[(2S)-2-10 aminopropanoyl]amino]propanoyl]amino]phenyl]methoxy]-4-(4-methylthiazol-5- yl)phenyl]ethyl]-4-hydroxy-pyrrolidine-2-carboxamide Int 209h (50 mg, 37.5 μmol, 96.6% yield) as a yellow solid. LCMS: MS (ESI) m/z 1330.4[M+H]⁺ Step I. Preparation of WVL-6 332 15077.006WO1 To a solution of Int 209h (20 mg, 15.0 μmol, 1 eq) in DMF (0.5 mL) were added DIEA (5.8 mg, 45.1 μmol, 7.85 μL, 3 eq) and 64-(3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-2-fluoro- 6-(3-oxo-3-(2,3,5,6-tetrafluorophenoxy)propyl)phenoxy)- 5 3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosamethyl- 4,7,10,13,16,19,22,25,28,31,34,37,40,43,46,49,52,55,58,61-icosaoxo- 3,6,9,12,15,18,21,24,27,30,33,36,39,42,45,48,51,54,57,60-icosaazatetrahexacontanoic acid Int L (30.5 mg, 15.8 μmol, 1.05 eq). The reaction mixture was stirred at 25°C for 1 hr. The mixture was acidified with TFA (PH=5~6) and filtered, then the filtrate was purified by pre-HPLC (TFA10 condition; column: Phenomenex Luna C1875*30mm*3um;mobile phase: [H₂O (0.1% TFA)- ACN];gradient:15%-50% B over 8.0 min ) to give WVL-6 (20.5 mg, 6.39 μmol, 42.4% yield, 96.5% purity) as a yellow solid.¹H NMR (400 MHz, DMSO-d6) δ9.94 (s, 1H), 8.97 (s, 1H), 8.84 (s, 1H), 8.45-8.43 (m, 1H), 8.19-8.10 (m, 2H), 8.07-7.98 (m, 1H), 7.79-7.58 (m, 7H), 7.45- 7.39 (m, 2H), 7.29-7.26 (m, 1H), 7.24 (s, 2H), 7.19 -7.08 (m, 2H), 7.08-6.97 (m, 4H), 5.74-5.59 15 (m, 1H), 5.34-5.05 (m, 5H), 5.00-4.97(m, 1H), 4.85-4.78 (m, 1H), 4.74-4.65 (m, 2H), 4.36-3.92 (m, 46H), 3.83-3.59 (m, 10H), 3.21-3.12 (m, 4H), 2.96-2.69 (m, 64H), 2.40 (s, 3H), 2.11-1.86 (m, 4H), 1.80-1.66 (m, 5H), 1.46 (s, 6H), 1.32-1.29 (m, 7H), 1.22-1.15 (m, 3H), 1.04 -1.03 (m, 3H), 0.67-0.65 (m, 3H). LCMS: MS (ESI) m/z 1550.4[1/2M+H]⁺ 333 15077.006WO1 The following compounds were prepared in a manner similar to that described for WVL- 6 using the appropriate compound as starting material. Example 101 Generation of Degrader Antibody Conjugate (DAC) compositions through 5 Reduction of Native Disulfide Bonds of Non-Engineered Antibodies 334 15077.006WO1 An antibody (e.g., a mAb at 3-8 mg/mL in PBS) is exchanged into HEPES (100 mM, pH 7.0, 1 mM DTPA) via molecular weight cut-off centrifugal filtration (Millipore, 30 kDa). The resultant solution is transferred to a tared 50 mL conical tube. The protein concentration is determined to be 3-8 mg/mL by A280. To the protein solution is added a reducing agent such as 5 tris(2-carboxyethyl)phosphine, TCEP or dithiothreitol, DTT (2.0-10.0 equivalents, 1 mM stock) at room temperature and the resultant mixture is incubated at 37 °C for 30-90 minutes, with gentle shaking. Upon being cooled to room temperature, a stir bar is added to the reaction tube. Next, a target protein binder and VHL ligand linker ((TPI-VHL)-L) compound(5.0-10.0 equivalents, 10 mM DMSO) is added dropwise. The resultant reaction mixture is allowed to stir 10 at ambient temperature for 30-60 minutes, at which point N-ethyl maleimide (3.0 equivalents, 100 mM DMA) is added. After an additional 15 minutes of stirring, N-acetylcysteine (6.0-11.0 equivalents, 50 mM HEPES) is added. The crude antibody conjugate is then exchanged into PBS and purified by preparative SEC (e.g. HiLoad 26/600, Superdex 200 pg) using PBS as the mobile phase. The pure fractions of the DAC are concentrated via molecular weight cut-off 15 centrifugal filtration (Millipore, 30 kDa), sterile filtered, and transferred to 15 mL conical tubes. The concentration was determined by UV-Vis and BCA, the monomer purity was determined by aSEC-HPLC and the final DAR was determined by HIC-HPLC and reduced LC-MS. The level of endotoxin determined by MCS analysis and free drug % was determined by RP-HPLC. Example 102 Preparation of Cysteine-engineered, Degrader Antibody Conjugate (DAC) 20 Approximately 50 mg of monoclonal antibody, mAb was pipetted into a 15-ml Eppendorf-type tube. Next, 2 mM EDTA was added from a 0.5 M stock solution and 50 mM DTT from a 1 M stock solution. The mixture was incubated overnight at room temperature. The reduction of the mAb was monitored using RP-HPLC and LC-MS. After confirming the reduction, DTT was removed using a 30K MWCO (molecular weight cut off) filter (6 washes 25 using 1x PBS). The concentration of the mAb was then checked by NanoDrop. The reduced mAb was treated with 20 to 25 equivalents of DHAA, and the reaction mixture was incubated at room temperature for 90 to 120 minutes. The reoxidation step was monitored using RP-HPLC. Then, DHAA was removed using 30K MWCO filters (3 times with 1XPBS). The concentration of the mAb was then checked by NanoDrop. 30 To 4 mg of de-capped antibody was added EDTA (to 2 mM final concentration) following the addition of a 20% organic co-solvent (propylene glcol), resulting in a final reaction concentration of 4-5 mg/ml. Next, 6 equivalents of a 10 mM Bcl-xL-VHL linker compound (BXV-L) was added to the reaction mixture, which was then incubated at room temperature for 90 minutes. The conjugation reaction was monitored using LC-MS. Crude 335 15077.006WO1 conjugate was purified via FPLC, SEC Superdex™ 200 (Cytiva) 24ml column; Running/Elution Buffer:1XPBS PH 7.0. Purified fractions were then collected, and the buffer was exchanged twice with 1XPBS using 30K MWCO filters. Example 103 In vitro cell proliferation assay 5 One day prior to treatment with a target protein binder and VHL ligand (TPI-VHL) compounds or a Degrader Antibody Conjugate (DAC), cells were plated in white 96-well plates. Compounds or DACs were added in a serial-dilution manner reaching the indicated concentrations. Optionally, 10 nM paclitaxel (standard-of-care chemotherapy agent in breast cancer) was added to the wells at this stage. Plates were incubated at 37 degrees for 96h (ADCs) 10 or 72h (compounds) whereafter a 1:10 dilution of Cell Titer Glow was used to measure relative cell numbers after 10min of incubation at room temperature. Example 104 Degradation assay evaluated with JESS (western blot)® One day prior to treatment cells were seeded in 6- or 12-well plates at a confluency of 70%. TPI-VHL compounds or Degrader Antibody Conjugates (DAC) were added in the 15 indicated concentrations and cells incubated at 37°C for indicated duration (6h, 16h, 24h). After washing the cells with ice cold PBS, RIPA lysis buffer containing protease and phosphatase inhibitors was added and cells were scraped off the plates. After vigorous vortexing, lysates were incubated on ice for 10min and subsequently centrifuged at 12000 rpm for 12min at 4°C. Supernatants were transferred to a new tube and used for analysis on the JESS SimpleWestern™ 20 Automated Western Blot System (ProteinSimple). 336 15077.006WO1 Table 4a Degradation Potency testing of RIPK2-VHL compounds (RV) from Table 1a Table 4b1 Degradation Potency and Cell Viability Assay testing of Bcl-xL-VHL compounds (BXV) from Tables 1b1 and 1b2 337 15077.006WO1 338 15077.006WO1 339 15077.006WO1 Table 4c Degradation Potency and Cell Viability Assay testing of KRAS-VHL compounds (KV) from Table 1c Table 4d Degradation Potency and Cell Viability Assay testing of WEE1-VHL compounds (WV) from Table 1d 340 15077.006WO1 341 15077.006WO1 Table 5 a Degradation Potency testing of RIPK2-VHL Degrader Antibody Conjugates (DAC-RV) from Table 3a Table 5b1 Degradation Potency and Cell Viability Assay testing of BCL-xL-VHL Degrader Antibody Conjugates (DAC-BXV) from Table 3b1 342 15077.006WO1 Table 5b2 Degradation Potency and Cell Viability Assay testing of BCL-xL-VHL Degrader Antibody Conjugates (DAC-BXV) from Table 3b2 343 15077.006WO1 344 15077.006WO1 345 15077.006WO1 Table 5c Degradation Potency and Cell Viability Assay testing of KRAS-VHL Degrader Antibody Conjugates (DAC-KV) from Table 3c 346 15077.006WO1 Table 5d1 Degradation Potency and Cell Viability Assay testing of WEE1-VHL Degrader Antibody Conjugates (DAC-WV) from Table 3d1 347 15077.006WO1 All references, including publications, patent applications, and patents, cited herein are hereby incorporated by reference to the same extent as if each reference were individually and specifically indicated to be incorporated by reference and were set forth in its entirety herein. 5 348

Claims

15077.006WO1 CLAIMS 1. A degrader antibody conjugate composition comprising a target protein binder and VHL ligand moiety (TPI-VHL) covalently attached to an antibody by an antibody linker, wherein the antibody binds to a tumor-associated antigen or cell-surface receptor. 2. The degrader antibody conjugate composition of claim 1 wherein the antibody has a modified Fc region. 3. The degrader antibody conjugate composition of claim 2 wherein the antibody has Fc mutations selected from: (i) LALAPA (L234A/L235A/P329A); (ii) LALAPG (L234A/L235A/P329G); (iii) LALASKPA (L234A, L235A, S267K, P329A); (iv) LALAPA-YTE (L234A/L235A/P329A-M252Y/S254T/T256E); (v) LALAPG-YTE (L234A/L235A/P329G-M252Y/S254T/T256E); and (vi) LALASKPA-YTE (L234A, L235A, S267K, P329A- M252Y/S254T/T256E), according to EU numbering. 4. The degrader antibody conjugate composition of claim 1, wherein the antibody linker is covalently attached to a cysteine amino acid of the antibody. 5. The degrader antibody conjugate composition of claim 2, wherein the antibody is a cysteine-engineered antibody. 6. The degrader antibody conjugate composition of claim 5 wherein the antibody has one engineered cysteine mutation site selected from heavy-chain E152C, S239C, K246C and S375C, numbered according to the EU system. 7. The degrader antibody conjugate composition of claim 5 wherein the antibody has two or three engineered cysteine mutation sites selected from heavy-chain E152C, S239C, K246C and S375C, numbered according to the EU system. 8. The degrader antibody conjugate composition of claim 5 wherein the cysteine- mutant antibody comprises a heavy chain cysteine mutation in a sequence selected from the group consisting of: 349

15077.006WO1 according to the EU system. 9. The degrader antibody conjugate composition of claim 5 wherein the cysteine- mutant antibody comprising one, two, or three engineered cysteine mutation sites is selected from (i) to (xvi): (i) HC S239C; (ii) HC K246C; (iii) HC S375C; (iv) HC E152C; (v) HC S239C and K246C; (vi) HC S239C and S375C; (vii) HC S239C and E152C; (viii) HC K246C and S375C; (ix) HC K246C and E152C; (x) HC S375C and E152C; (xi) HC S239C, K246C, and S375C; (xii) HC S239C, K246C, and E152C; (xiii) HC S239C, S375C, and E152C; (xiv) HC K246C, S375C, and E152C; (xv) HC S239C, S375C, and E152C; (xvi) HC K246C, S375C, and E152C. 10. The degrader antibody conjugate composition of claim 1 wherein the target protein binder of the TPI-VHL binds to RIPK2, KRAS, Bcl-XL, or WEE1. 11. The degrader antibody conjugate composition of any one of claims 1 to 10 having Formula I: Ab−[L−(TPI−Sp−VHL)]_p I or a pharmaceutically acceptable salt thereof, 350

15077.006WO1 wherein: Ab is the antibody; L is the antibody linker; TPI−Sp−VHL is a moiety comprising a target protein binder TPI and a VHL ligand wherein the TPI is covalently attached to the VHL ligand by a spacer unit Sp; and p is an integer from 1 to 12; the TPI, the VHL ligand, or the spacer unit Sp is attached to the antibody linker L; and the VHL ligand has Formula Ia: wherein A is a cyclic structure selected from C₃−C₂₀ carbocyclyl, C₆−C₂₀ aryl, C₁−C₂₀ heteroaryl, and C₂-C₂₀ heterocyclyl, each of which are substituted with one or more groups independently selected from H, F, Cl, Br, I, −CN, −NO2, C1−C12 alkyl, C2−C12 alkenyl, C2−C12 alkynyl, C₁−C₁₂ heteroalkyl, −(C₁−C₁₂ heteroalkyldiyl)−(C₆−C₂₀ aryl), −(C₁−C₁₂ heteroalkyldiyl)−(C₆−C₂₀ aryldiyl)−(C₁−C₁₂ heteroalkyl), −(C₁−C₁₂ heteroalkyldiyl)−(C₆−C₂₀ aryldiyl)−(C₂-C₂₀ heterocyclyl), (C₁−C₆ alkyldiyl)−(C₆−C₂₀ aryl), −(C₁−C₆ alkyldiyl)−NR^aR^b, −(C₁−C₆ alkyldiyl)−OR^a, (C₁−C₆ alkyldiyl)−(C₃−C₂₀ carbocyclyl), (C₁-C₆ alkyldiyl)−(C₂-C₂₀ heterocyclyl), (C₁−C₆ alkyldiyl)−(C₁−C₂₀ heteroaryl), C₆−C₂₀ aryl, C₃−C₂₀ carbocyclyl, C₂−C₂₀ heterocyclyl, C₁−C₂₀ heteroaryl, −C(=NH)NH(OH), −C(=NH)NH₂, −C(=O)NR^aR^b, −C(=O)NR^a−NR^aR^b, −C(=O)NH(C₁-C₆ alkyldiyl)−NR^aR^b, −C(=O)OR^a, −NR^aR^b, −NR^aS(O)₂R^a, −NO₂, −OR^a, −OC(=O)R^a, −SR^a, −S(O)R^a, −S(O)₂R^a, −S(O)₂NR^a, and −S(O)₃H; R^a is independently selected from H, C1−C6 alkyl, phenyl, and benzyl, wherein phenyl and benzyl are optionally substituted with one or more groups independently selected from the group consisting of F, Cl, −CN, C₁−C₁₂ alkyl, C₂−C₁₂ alkenyl, and C₂−C₁₂ alkynyl; or where two R^a groups form a five-, six-, or seven-membered heteroaryl or heterocyclyl ring; 351

15077.006WO1 R^b is independently selected from H, OH, C1−C6 alkyl, phenyl, and benzyl, wherein phenyl and benzyl are optionally substituted with one or more groups independently selected from the group consisting of F, Cl, −CN, C₁−C₁₂ alkyl, C₂−C₁₂ alkenyl, and C₂−C₁₂ alkynyl; R^c is selected from OH and −SO2F; R^d is selected from H and F; Cyc is a ring structure selected from the group consisting of C₃−C₂₀ carbocyclyl, C₆−C₂₀ aryl, C₂-C₂₀ heterocyclyl, C₁−C₂₀ heteroaryl, and −(C₁−C₂₀ heteroaryldiyl)−(C₁−C₁₂ heteroalkyl); n is 0 or 1; R¹ is selected from the group consisting of H, C1−C12 alkyl, and C1−C12 heteroalkyl; R² is selected from the group consisting of H, C1−C12 alkyl, and C1−C12 heteroalkyl, or where R² forms a five- to ten-membered aryl, carbocyclyl, heteroaryl or heterocyclyl ring with A; X¹ is selected from the group consisting of H, −NHC(=O)−, (C₃−C₂₀ carbocyclyl)− C(=O)NH−, C₁−C₁₂ heteroalkyl, and C₁−C₂₀ heteroaryl; or where X¹ forms a five-, six-, or seven-membered heteroaryl or heterocyclyl ring with R¹; Sp is selected from the group consisting of a bond, O, NH, C₁−C₁₂ alkyldiyl, C₁−C₆₀ heteroalkyldiyl, C₃−C₂₀ carbocyclyldiyl, C₂-C₂₀ heterocyclyldiyl, C₆-C₂₀ aryldiyl, C₁−C₄₀ heteroaryldiyl, −(C₃−C₂₀ carbocyclyldiyl)−(C₁−C₆₀ heteroalkyldiyl)−, −(C₃−C₂₀ carbocyclyldiyl)−(C₁−C₁₂ alkyldiyl)−, −(C₂-C₂₀ heterocyclyldiyl)−(C₁−C₆₀ heteroalkyldiyl)−, −(C₂-C₂₀ heterocyclyldiyl)−(C₁−C₁₂ alkyldiyl)−, −(C₂-C₂₀ heterocyclyldiyl)−(C₂-C₂₀ heterocyclyldiyl)−, −(C₁−C₂₀ heteroaryldiyl)−(C₃−C₂₀ carbocyclyldiyl)−(C₁−C₆₀ heteroalkyldiyl)−, a solubilizing unit, and combinations thereof, where the solubilizing unit is selected from polyglycine, polysarcosine, polyethyleneoxy (PEG), a glycoside, C1−C60 heteroalkyldiyl, and combinations thereof; L is the antibody linker; one of A, R^a, R^b, Cyc, R¹, R², and X¹ is attached to Sp; and one of A, R^a, R^b, Cyc, R¹, R², X¹ and Sp is attached to L; where each alkyl, alkyldiyl, alkenyl, alkenyldiyl, alkynyl, alkynyldiyl, heteroalkyl, heteroalkyldiyl, aryl, aryldiyl, carbocyclyl, carbocyclyldiyl, heterocyclyl, heterocyclyldiyl, heteroaryl, and heteroaryldiyl is independently substituted with one or more groups selected from H, F, Cl, Br, I, −CN, −CH3, −CH2CH3, −CH=CH2, −C^CH, −C^CCH3, −C^CCH2NHCH3, −CH2CH2CH3, −CH(CH3)2, −CH2CH(CH3)2, −CH(CH2CH2), −CH(CH2CH2CH2), −CH2OH, − CH2OCH3, −CH2CH2OH, −C(CH3)2OH, −CH(OH)CH(CH3)2, −C(CH3)2CH2OH, − 352

15077.006WO1 CH2CH2SO2CH3, −CH2OP(O)(OH)2, −CH2F, −CHF2, −CF3, −CH2CF3, −CH2CHF2, − CH(CH3)CN, −C(CH3)2CN, −CH2CN, −CH2NH2, −CH2NHSO2CH3, −CH2NHCH3, − CH2N(CH3)2, −CO2H, −COCH3, −CO2CH3, −CO2C(CH3)3, −COCH(OH)CH3, −CONH2, − CONHCH3, −CON(CH3)2, −C(CH3)2CONH2, −NH2, −NHCH3, −N(CH3)2, −NHCOCH3, − N(CH3)COCH3, −NHS(O)2CH3, −N(CH3)C(CH3)2CONH2, −N(CH3)CH2CH2S(O)2CH3, − NHC(=NH)H, −NHC(=NH)CH3, −NHC(=NH)NH2, −NHC(=O)NH2, −NO2, =O, −OH, −OCH3, −OCH₂CH₃, −OCH₂CH₂OCH₃, −OCH₂CH₂OH, −OCH₂CH₂N(CH₃)₂, −OCH₂F, −OCHF₂, − OCF₃, −OP(O)(OH)₂, −S(O)₂N(CH₃)₂, −SCH₃, −S(O)₂CH₃, and −S(O)₃H. 12. The degrader antibody conjugate composition of claim 11 wherein Cyc is a five-, six-, or seven-membered ring structure selected from the group consisting of thiazole, triazole, phenyl, pyridine, pyrazine, pyridazine, and pyrimidine, substituted with one or more groups independently selected from H, F, Cl, Br, I, −CN, −CH₃, −CH₂CH₃, −CH=CH₂, −C^CH, − C^CCH₃, −C^CCH₂NHCH₃, −CH₂CH₂CH₃, −CH(CH₃)₂, −CH₂CH(CH₃)₂, −CH(CH₂CH₂), − CH(CH₂CH₂CH₂), −CH₂OH, −CH₂OCH₃, −CH₂CH₂OH, −C(CH₃)₂OH, −CH(OH)CH(CH₃)₂, − C(CH₃)₂CH₂OH, −CH₂CH₂SO₂CH₃, −CH₂OP(O)(OH)₂, −CH₂F, −CHF₂, −CF₃, −CH₂CF₃, − CH2CHF2, −CH(CH3)CN, −C(CH3)2CN, −CH2CN, −CH2NH2, −CH2NHSO2CH3, −CH2NHCH3, −CH2N(CH3)2, −CO2H, −COCH3, −CO2CH3, −CO2C(CH3)3, −COCH(OH)CH3, −CONH2, − CONHCH3, −CON(CH3)2, −C(CH3)2CONH2, −NH2, −NHCH3, −N(CH3)2, −NHCOCH3, − N(CH3)COCH3, −NHS(O)2CH3, −N(CH3)C(CH3)2CONH2, −N(CH3)CH2CH2S(O)2CH3, − NHC(=NH)H, −NHC(=NH)CH3, −NHC(=NH)NH2, −NHC(=O)NH2, −NO2, =O, −OH, −OCH3, −OCH2CH3, −OCH2CH2OCH3, −OCH2CH2OH, −OCH2CH2N(CH3)2, −OCH2F, −OCHF2, − OCF3, −OP(O)(OH)2, −S(O)2N(CH3)2, −SCH3, −S(O)2CH3, and −S(O)3H. 13. The degrader antibody conjugate composition of claim 11 wherein R¹ is C1−C12 alkyl. 14. The degrader antibody conjugate composition of claim 11 wherein R² is selected from the group consisting of H, C₁−C₁₂ alkyl, and C₁−C₁₂ heteroalkyl. 15. The degrader antibody conjugate composition of claim 14 wherein R² is selected from H, −CH₃, −CH₂OH, −CH₂CH₂OH, −CH₂OPO₂OH, −CH₂CH₂OPO₂OH, −CH₂COOH, −CH₂CH₂COOH, −CH₂CH₂CONHS(O)₂CH₃, and −CH₂CH₂CONHS(O)₂CH₂CH₂N(CH₃)₂.. 16. The degrader antibody conjugate composition of claim 11 wherein X¹ is C₁−C₂₀ heteroaryldiyl selected from the group consisting of triazole, isoxazole, and oxadiazole. 17. The degrader antibody conjugate composition of claim 11 wherein X¹ is − C(=O)NH−. 353

15077.006WO1 18. The degrader antibody conjugate composition of claim 11 wherein the VHL ligand has Formula Ib: . 19. The degrader antibody conjugate composition of claim 18 wherein the VHL ligand has Formula Ic: wherein: R³, R⁴, R⁵, and R⁶ are independently selected from the group consisting of H, F, Cl, Br, I, −CN, C₁−C₁₂ alkyl, C₂−C₁₂ alkenyl, C₂−C₁₂ alkynyl, C₁−C₁₂ heteroalkyl, −(C₁−C₁₂ heteroalkyldiyl)−(C₆−C₂₀ aryl), −(C₁−C₁₂ heteroalkyldiyl)−(C₆−C₂₀ aryldiyl)−(C₁−C₁₂ heteroalkyl), −(C₁−C₁₂ heteroalkyldiyl)−(C₆−C₂₀ aryldiyl)−(C₂-C₂₀ heterocyclyl), (C₁−C₆ alkyldiyl)−(C₆−C₂₀ aryl), −(C₁−C₆ alkyldiyl)−NR^aR^b, −(C₁−C₆ alkyldiyl)−OR^a, (C₁−C₆ alkyldiyl)−(C₃−C₂₀ carbocyclyl), (C₁-C₆ alkyldiyl)−(C₂-C₂₀ heterocyclyl), (C₁−C₆ alkyldiyl)−(C₁−C₂₀ heteroaryl), C₆−C₂₀ aryl, C₃−C₂₀ carbocyclyl, C₂−C₂₀ heterocyclyl, C₁−C₂₀ heteroaryl, −C(=NH)NH(OH), −C(=NH)NH2, −C(=O)NR^aR^b, −C(=O)NR^a−NR^aR^b, −C(=O)NH(C1-C6 alkyldiyl)−NR^aR^b, −C(=O)OR^a, −NR^aR^b, −NO2, −OR^a, −OC(=O)R^a, −SR^a, −S(O)R^a, −S(O)2R^a, −S(O)2NR^a, and −S(O)3H; 354

15077.006WO1 R^a is independently selected from H, C1−C6 alkyl, phenyl, and benzyl, wherein phenyl and benzyl are optionally substituted with one or more groups independently selected from the group consisting of F, Cl, −CN, C₁−C₁₂ alkyl, C₂−C₁₂ alkenyl, and C₂−C₁₂ alkynyl; or where two R^a groups form a five-, six-, or seven-membered heteroaryl or heterocyclyl group; R^b is independently selected from H, OH, C1−C6 alkyl, phenyl, and benzyl, wherein phenyl and benzyl are optionally substituted with one or more groups independently selected from the group consisting of F, Cl, −CN, C1−C12 alkyl, C2−C12 alkenyl, and C2−C12 alkynyl; or where R³ and R⁴ together form a five-membered or six-membered heteroaryl or heterocyclyl group comprising one or more heteroatoms independently selected from N, O, P and S; one of X¹, R¹, R², R³, R⁴, R⁵, R⁶, R^a, R^b, and Cyc is attached to the spacer unit; one of X¹, R¹, R², R³, R⁴, R⁵, R⁶, R^a, R^b, Cyc, and Sp is attached to L; and where each alkyl, alkyldiyl, alkenyl, alkenyldiyl, alkynyl, alkynyldiyl, heteroalkyl, heteroalkyldiyl, aryl, aryldiyl, carbocyclyl, carbocyclyldiyl, heterocyclyl, heterocyclyldiyl, heteroaryl, and heteroaryldiyl is independently and optionally substituted with one or more groups selected from F, Cl, Br, I, −CN, −CH₃, −CH₂CH₃, −CH=CH₂, −C^CH, −C^CCH₃, − C^CCH₂NHCH₃, −CH₂CH₂CH₃, −CH(CH₃)₂, −CH₂CH(CH₃)₂, −CH₂OH, −CH₂OCH₃, − CH₂CH₂OH, −C(CH₃)₂OH, −CH(OH)CH(CH₃)₂, −C(CH₃)₂CH₂OH, −CH₂CH₂SO₂CH₃, − CH2OP(O)(OH)2, −CH2F, −CHF2, −CF3, −CH2CF3, −CH2CHF2, −CH(CH3)CN, −C(CH3)2CN, − CH2CN, −CH2NH2, −CH2NHSO2CH3, −CH2NHCH3, −CH2N(CH3)2, −CO2H, −COCH3, − CO2CH3, −CO2C(CH3)3, −COCH(OH)CH3, −CONH2, −CONHCH3, −CON(CH3)2, − CONHS(O)2N(CH3)2, −C(CH3)2CONH2, −NH2, −NHCH3, −N(CH3)2, −NHCOCH3, − N(CH3)COCH3, −NHS(O)2CH3, −N(CH3)C(CH3)2CONH2, −N(CH3)CH2CH2S(O)2CH3, − NHC(=NH)H, −NHC(=NH)CH3, −NHC(=NH)NH2, −NHC(=O)NH2, −NO2, =O, −OH, −OCH3, −OCH2CH3, −OCH2CH2OCH3, −OCH2CH2OH, −OCH2CH2N(CH3)2, −OCH2F, −OCHF2, − OCF3, −OP(O)(OH)2, −S(O)2N(CH3)2, −SCH3, −S(O)2CH3, and −S(O)3H. 20. The degrader antibody conjugate composition of claim 19 wherein one or more of R³, R⁴, R⁵, and R⁶ are independently selected from F and OH. 21. The degrader antibody conjugate composition of claim 19 wherein X¹ forms a five-, six-, or seven-membered heteroaryl or heterocyclyl ring with R¹. 22. The degrader antibody conjugate composition of claim 11 wherein X¹ is attached to the spacer unit. 355

15077.006WO1 23. The degrader antibody conjugate composition of claim 11 wherein Sp is C1−C60 heteroalkyldiyl having the formula: −(CH2CH2X²)n−(CH2)m− where X² is independently selected from NH and O, m is an integer from 1 to 5, and n is an integer from 1 to 50. 24. The degrader antibody conjugate composition of claim 11 wherein Sp is −(C₃−C₂₀ carbocyclyldiyl)−(C₁−C₆₀ heteroalkyldiyl)− or C₁−C₆₀ heteroalkyldiyl. 25. The degrader antibody conjugate composition of claim 24 wherein Sp is selected from the group consisting of −S(O)2CH2CH2−, −S(O)2NHCH2CH2−, −S(O)2CH2CH2CH2−, − S(O)₂NHCH₂CH₂CH₂−, −S(O)₂CH₂CH₂CH₂O−, and −S(O)₂NHCH₂CH₂CH₂O−. 26. The degrader antibody conjugate composition of claim 11 wherein Sp has a formula selected from the group consisting of: 356

15077.006WO1 where the wavy lines indicate the points of attachment to the target protein binder TPI and to the VHL ligand. 27. The degrader antibody conjugate composition of claim 11 wherein the target protein binder TPI binds to Bcl-xL and is selected from the structures: 357

15077.006WO1 wherein: R¹⁰ is a bicyclic C₁−C₂₀ heteroaryl substituted with one or more groups independently selected from H, F, Cl, −CH₃, −CN, −NO₂, −OH, and −OCH₃; R¹¹ is selected from H, −(C6−C20 aryl), −(C1−C20 heteroaryldiyl)−(C1-C12 alkyl), −(C1−C20 heteroaryldiyl)−(C1-C12 alkyldiyl)−(C3−C20 carbocyclyldiyl)−(C1−C60 heteroalkyl), and −(C1−C20 heteroaryldiyl)−(C3−C20 carbocyclyldiyl)−(C1−C60 heteroalkyl); R¹² is selected from −(C1−C20 heteroaryldiyl)−(C1-C12 alkyldiyl)−(C3−C20 carbocyclyldiyl)−(C1−C60 heteroalkyldiyl)−*; R¹³ is selected from −(C1−C60 heteroalkyldiyl)−*; and the asterisk * indicates the attachment to the spacer Sp. 28. The degrader antibody conjugate composition of claim 27 wherein R¹⁰ is selected from: , substituted with one or more groups independently selected from H, F, Cl, −CH3, −CN, −NO2, −OH, and −OCH3. 29. The degrader antibody conjugate composition of claim 28 wherein R¹⁰ is: . 358

15077.006WO1 30. The degrader antibody conjugate composition of claim 11 wherein the target protein binder TPI binds to Bcl-xL and is selected from the structures: 359

15077.006WO1 where the wavy line indicates the point of attachment to the spacer unit Sp. 31. The degrader antibody conjugate composition of claim 11 wherein the target protein binder TPI binds to KRAS and has Formula Id: wherein: X², X³, and X⁴ are independently selected from the group consisting of C(R^aR^b), O, S, and NR^a, 360

15077.006WO1 or where: (i) X² and X⁴, (ii) X³ and X⁴, or (iii) X² and X³ can form a 3-6 membered carbocyclic ring; the dotted line indicates an optional double bond; Y¹ is selected from the group consisting of CH, N, O, S; Y² is selected from the group consisting of N, O, S; Y³ is selected from the group consisting of N, O, S; R¹⁴ is selected from the group consisting of H, C1−C6 alkyl, C2−C12 alkenyl, and C2−C12 alkynyl, substituted with one or more groups independently selected from the group consisting of H, F, Cl, −CN, −NO₂, −OH, and −CN; or where R¹⁴ and Y¹ form a six-membered ring, optionally-substituted with one or more groups selected from F, spiro-cyclopropyl, C1−C12 heteroalkyl, and C1−C12 alkyl; R¹⁵ is selected from the group consisting of −O−(C₁−C₁₂ heteroalkyldiyl)−(C₂-C₂₀ heterocyclyl), −O−(C₁−C₁₂ alkyldiyl)−(C₂-C₂₀ heterocyclyl), −(C₁−C₁₂ heteroalkyldiyl)−(C₂-C₂₀ heterocyclyl), −(C₁−C₁₂ alkyldiyl)−(C₂-C₂₀ heterocyclyl), −(C₂-C₂₀ heterocyclyl)−(C₂-C₂₀ heterocyclyl), and C₂-C₂₀ heterocyclyl, where at least one of the heterocyclyl ring atoms comprise one or more basic nitrogens, and the heterocyclyl group is substituted with one or more groups independently selected from H, F, Cl, =CH2, C1−C6 alkyl, −C(=NH)NH(OH), −C(=NH)NH2, −C(=O)NR^aR^b, −C(=O)NR^a−NR^aR^b, −C(=O)NH−(C1-C6 alkyldiyl)−NR^aR^b, −C(=O)OR^a, −NR^aR^b, −NO2, −OR^a, −OC(=O)R^a, −SR^a, −S(O)R^a, −S(O)2R^a, −S(O)2NR^a, and −S(O)3H; R^a is independently selected from H, C1−C6 alkyl, phenyl, and benzyl, wherein phenyl and benzyl are substituted with one or more groups independently selected from the group consisting of H, F, Cl, −CN, −NO2, −OH, C1−C12 alkyl, C2−C12 alkenyl, and C2−C12 alkynyl; or where two R^a groups form a five-, six-, or seven-membered heteroaryl or heterocyclyl ring; R^b is independently selected from H, OH, C1−C6 alkyl, phenyl, and benzyl, wherein phenyl and benzyl are substituted with one or more groups independently selected from the group consisting of H, F, Cl, −CN, C1−C12 alkyl, C2−C12 alkenyl, and C2−C12 alkynyl; R¹⁶ is selected from the group consisting of −(C1−C20 heteroaryldiyl)−*, −(C1−C20 heteroaryldiyl)−C(=O)NR^a−*, −(C1−C20 heteroaryldiyl)−(C1−C6 alkyldiyl)−C(=O)NR^a−*, −(C1−C20 heteroaryldiyl)−(C1−C6 alkyldiyl)−*, −(C6−C20 aryldiyl)−*, −(C6−C20 aryldiyl)−C(=O)NR^a−*, −(C₆−C₂₀ aryldiyl)−(C₁−C₆ alkyldiyl)−C(=O)NR^a−*, −(C₁−C₂₀ heteroaryldiyl)−(C₁−C₆ alkyldiyl)−(C₁−C₂₀ heteroaryldiyl)−*, −(C₁−C₂₀ heteroaryldiyl)−(C₁−C₆ 361

15077.006WO1 heteroalkyldiyl)−(C1−C20 heteroaryldiyl)−*, −(C1−C20 heteroaryldiyl)−(C1−C6 alkyldiyl)−(C1−C20 heterocyclyl)−*, −(C1−C20 heteroaryldiyl)−(C1−C6 heteroalkyldiyl)−(C1−C20 heterocyclyl)−*, −O−(C1−C20 heteroaryldiyl)−*, and −O−(C1−C20 heteroaryldiyl)−C(=O)NR^a−*; where the asterisk is the attachment site to the spacer unit Sp. 32. The degrader antibody conjugate composition of claim 31 wherein the target protein binder TPI has a formula selected from the group consisting of: where the wavy line indicates the point of attachment to the spacer unit Sp. 33. The degrader antibody conjugate composition of claim 11 wherein the target protein binder TPI binds to WEE1 and has Formula Ie: Ie wherein: R¹⁷ is selected from the group consisting of C₁−C₆ alkyl, C₂−C₆ alkenyl, and C₂−C₆ alkynyl, each of which are substituted with one or more groups independently selected from H, F, Cl, −CH3, −CN, −NO2, −OH, and −OCH3; R¹⁸ is C₁−C₂₀ heteroaryl substituted with one or more groups independently selected from H, F, Cl, −CN, −NO2, −OH, −OCH3, C1−C6 alkyl, and C1−C20 heteroalkyl; R¹⁹ is selected from the group consisting of H and C₁−C₆ alkyl; and R²⁰ is C₁−C₆ alkyldiyl attached to the spacer Sp; or where R¹⁹ and R²⁰ form a five-, six-, or seven-membered heterocycle attached to the spacer Sp; 362

15077.006WO1 where each alkyl, alkyldiyl, alkenyl, alkynyl, alkynyldiyl, heteroalkyl, heterocyclyl, and heteroaryl is independently and optionally substituted with one or more groups selected from F, Cl, Br, I, −CN, −CH₃, −CH₂CH₃, −CH=CH₂, −C^CH, −C^CCH₃, −C^CCH₂NHCH₃, − CH₂CH₂CH₃, −CH(CH₃)₂, −CH₂CH(CH₃)₂, −CH₂OH, −CH₂OCH₃, −CH₂CH₂OH, −C(CH₃)₂OH, −CH(OH)CH(CH₃)₂, −C(CH₃)₂CH₂OH, −CH₂CH₂SO₂CH₃, −CH₂OP(O)(OH)₂, −CH₂F, −CHF₂, −CF₃, −CH₂CF₃, −CH₂CHF₂, −CH(CH₃)CN, −C(CH₃)₂CN, −CH₂CN, −CH₂NH₂, − CH₂NHSO₂CH₃, −CH₂NHCH₃, −CH₂N(CH₃)₂, −CO₂H, −COCH₃, −CO₂CH₃, −CO₂C(CH₃)₃, − COCH(OH)CH₃, −CONH₂, −CONHCH₃, −CON(CH₃)₂, −C(CH₃)₂CONH₂, −NH₂, −NHCH₃, − N(CH3)2, −NHCOCH3, −N(CH3)COCH3, −NHS(O)2CH3, −N(CH3)C(CH3)2CONH2, − N(CH₃)CH₂CH₂S(O)₂CH₃, −NHC(=NH)H, −NHC(=NH)CH₃, −NHC(=NH)NH₂, − NHC(=O)NH2, −NO2, =O, −OH, −OCH3, −OCH2CH3, −OCH2CH2OCH3, −OCH2CH2OH, − OCH2CH2N(CH3)2, −OCH2F, −OCHF2, −OCF3, −OP(O)(OH)2, −S(O)2N(CH3)2, −SCH3, − S(O)2CH3, and −S(O)3H. 34. The degrader antibody conjugate composition of claim 33 wherein R¹⁷ is allyl. 35. The degrader antibody conjugate composition of claim 33 wherein R¹⁸ is pyridyl substituted with C₁−C₂₀ heteroalkyl. 36. The degrader antibody conjugate composition of claim 33 wherein R¹⁹ and R²⁰ form a piperidyl ring attached to the spacer Sp. 37. The degrader antibody conjugate composition of claim 33 wherein Formula Ie has the structure: where the asterisk is the attachment site to the spacer unit Sp. 38. The degrader antibody conjugate composition of claim 11 wherein L comprises a solubilizing unit selected from polyglycine, polysarcosine, polyethyleneoxy (PEG), and a glycoside, or combinations thereof. 39. The degrader antibody conjugate composition of claim 38 wherein the solubilizing unit is monovalent and the terminus of the solubilizing unit is a group selected from an amino acid, amino, hydroxyl, hydrogen, carboxylic acid, glycerol, or a sugar such as pentaerythritol, maltitol, sorbitol, xylitol, erythritol, isomalt, or combinations thereof. 363

15077.006WO1 40. The degrader antibody conjugate composition of claim 38 wherein the solubilizing unit and the terminus of the solubilizing unit covalently attached to Str are selected from the structures: ; ; ; ; 364

15077.006WO1 wherein * indicates the point of attachment to Str. 41. The degrader antibody conjugate composition of claim 11 wherein L has the structure: 365

15077.006WO1 wherein: L¹ is selected from a bond, C₁−C₁₂ alkyldiyl, and C₁−C₄₀ heteroalkyldiyl; L^1a is selected from an amino acid, amino, hydroxyl, halide, hydrogen, carboxylic acid, glycerol, or a sugar such as pentaerythritol, maltitol, sorbitol, xylitol, erythritol, isomalt, or combinations thereof; o is 0, 1, or 2; L² is selected from a bond, C1−C12 alkyldiyl, and C1−C40 heteroalkyldiyl; * indicates the point of attachment to a cysteine thiol of Ab; and ** indicates the point of attachment to the TPI-VHL moiety. 42. The degrader antibody conjugate composition of claim 41 wherein L¹ is a bond and L^1a is F. 43. The degrader antibody conjugate composition of claim 11 wherein L has the formula: −Str−(PEP)_y−(IM)_z− wherein: Str is a stretcher unit covalently attached to the antibody; PEP is a protease-cleavable, peptide unit covalently attached to Str and IM or the TPI- VHL moiety; IM is an immolative unit covalently attached to the TPI-VHL moiety; y is 0 or 1; and z is 0 or 1. 44. The degrader antibody conjugate composition of claim 43 wherein L is a branched linker and Str is covalently attached to: (i) the antibody; and (ii) a solubilizing unit comprising a group selected from polyglycine, polysarcosine, polyethyleneoxy (PEG), and a glycoside, or combinations thereof, wherein the terminus of the solubilizing unit is a group selected from an amino acid, amino, hydroxyl, hydrogen, carboxylic acid, glycerol, or a sugar such as pentaerythritol, maltitol, sorbitol, xylitol, erythritol, isomalt, or combinations thereof. 45. The degrader antibody conjugate composition of claim 43 wherein L is a branched linker and PEP is covalently attached to: (i) Str and IM or the TPI-VHL moiety; and (ii) a solubilizing unit comprises a group selected from polyglycine, polysarcosine, 366

15077.006WO1 polyethyleneoxy (PEG), and a glycoside, or combinations thereof, wherein the terminus of the solubilizing unit is a group selected from an amino acid, amino, hydroxyl, hydrogen, carboxylic acid, glycerol, or a sugar such as pentaerythritol, maltitol, sorbitol, xylitol, erythritol, isomalt, or combinations thereof. 46. The degrader antibody conjugate composition of claim 43 wherein L is a branched linker and IM is covalently attached to: (i) the TPI-VHL moiety; and (ii) a solubilizing unit comprises a group selected from polyglycine, polysarcosine, polyethyleneoxy (PEG), and a glycoside, or combinations thereof, wherein the terminus of the solubilizing unit is a group selected from an amino acid, amino, hydroxyl, hydrogen, carboxylic acid, glycerol, or a sugar such as pentaerythritol, maltitol, sorbitol, xylitol, erythritol, isomalt, or combinations thereof. 47. The degrader antibody conjugate composition of claim 43 wherein Str has the structure: wherein: * indicates the point of attachment to a cysteine thiol of Ab; ** indicates the point of attachment to PEP or to the TPI-VHL moiety; R^e is selected from the group consisting of C1-C12 alkyldiyl, C1-C12 alkyldiyl-C(=O), C1- C12 alkyldiyl−NH, (CH2CH2O)r, (CH2CH2O)r−C(=O), (CH2CH2O)r-CH2, C1−C12 heteroalkyldiyl, C6−C20 aryldiyl, (C6−C20 aryldiyl)−(C1−C12 alkyldiyl), and (C6−C20 aryldiyl)− (C1−C12 heteroalkyldiyl); r is an integer ranging from 1 to 10; and alkyldiyl, heteroalkyldiyl, and aryldiyl are independently and optionally substituted with one or more groups selected from F, Cl, −CN, −NH₂, −CH₂NH₂, −OH, −OCH₃, −OCH₂CH₃, − OCH₂CH₂OCH₃, −OCH₂CH₂OH, −OCH₂CH₂N(CH₃)₂, −OCH₂F, −OCHF₂, −OCF₃, − OP(O)(OH)₂, −S(O)₂N(CH₃)₂, −SCH₃, −S(O)₂CH₃, −S(O)₃H, and a solubilizing unit. 48. The degrader antibody conjugate composition of claim 47 wherein R^e is selected from −(CH₂)₅−, −CH₂−, and −CH₂CH₂−. 49. The degrader antibody conjugate composition of claim 47 wherein R^e is selected from C6−C20 aryldiyl, (C6−C20 aryldiyl)−(C1−C12 alkyldiyl), and (C6−C20 aryldiyl)−(C1−C12 heteroalkyldiyl). 367

15077.006WO1 50. The degrader antibody conjugate composition of claim 43 wherein Str is selected from the structure: wherein: L¹ is selected from a bond, C1−C12 alkyldiyl, C1−C60 heteroalkyldiyl, and a solubilizing unit; L^1a is selected from an amino acid, amino, hydroxyl, hydrogen, carboxylic acid, glycerol, or a sugar such as pentaerythritol, maltitol, sorbitol, xylitol, erythritol, isomalt, or combinations thereof; o is 0, 1, or 2; L² is selected from a bond, C1−C12 alkyldiyl, C1−C60 heteroalkyldiyl, and a solubilizing unit; * indicates the point of attachment to a cysteine thiol of Ab; and ** indicates the point of attachment to PEP or to the TPI-VHL moiety. 51. The degrader antibody conjugate composition of claim 50 wherein L¹ or L² comprise a solubilizing unit selected from polyglycine, polysarcosine, polyethyleneoxy (PEG), and a glycoside, or combinations thereof. 52. The degrader antibody conjugate composition of claim 50 wherein L¹ or L² is selected from (N(CH3)CH2C(=O))q, (N(CH3)CH2CH2C(=O))q, N(CH₃)CH₂CH₂OCH₂CH₂C(=O))_q, (CH₂CH₂O)_q, (CH₂CH₂O)_q−C(=O), and (CH₂CH₂O)_q-CH₂, where q is an integer from 2 to 20. 53. The degrader antibody conjugate composition of claim 43 wherein n is 1, m is 1, and PEP-IM has the formula: wherein * indicates the point of attachment to Str and ** indicates the point of attachment to the TPI-VHL moiety; AA is independently selected from a natural or unnatural amino acid side chain, or one or more of AA, and an adjacent nitrogen atom form a 5-membered ring proline amino acid; 368

15077.006WO1 Cyc¹ is selected from C6-C20 aryldiyl and C1-C20 heteroaryldiyl, optionally substituted with one or more groups selected from F, Cl, NO₂, −OH, −OCH₃, a C-glycoside, and glucuronic acid having the structure: R⁷ is selected from the group consisting of −CH(R⁸)O−, −CH2−, −CH2N(R⁸)CH(R⁸)−, − CH(R⁸)OC(=O)−, −CH(R⁸)OC(=O)N(R⁸)CH(R⁸)−, −CH(R⁸)OP(=O)2OCH(R⁸)−, and − CH(R⁸)OC(=O)N(R⁸)−(C₁-C₆ alkyldiyl)−N(R⁸)C(=O)OCH(R⁸)−; R⁸ is selected from H, C₁-C₆ alkyl, C(=O)−C₁-C₆ alkyl, and −C(=O)N(R⁹)₂; R⁹ is independently selected from the group consisting of H, C₁-C₁₂ alkyl, and − (CH₂CH₂O)_n−(CH₂)_m−OH, where m is an integer from 1 to 5, and n is an integer from 2 to 50, or two R⁹ groups together form a 5- or 6-membered heterocyclyl ring; y¹ is an integer from 2 to 12; and z¹ is 0 or 1. 54. The degrader antibody conjugate composition of claim 43 wherein IM is selected from the formulae: 369

15077.006WO1 wherein: * indicates the point of attachment to PEP; and ** indicates the point of attachment to the TPI-VHL moiety. 55. A compound comprising a target protein binder (TPI) covalently attached to a VHL ligand by a spacer unit (Sp) having Formula II: TPI−Sp−VHL II or a pharmaceutically acceptable salt, stereoisomer, mixture of stereoisomers, tautomer, or isotopic analog thereof, wherein: TPI binds to RIPK2, KRAS, Bcl-XL, or WEE1; VHL ligand has Formula IIa: 370

15077.006WO1 IIa wherein A is a cyclic structure selected from C₃−C₂₀ carbocyclyl, C₆−C₂₀ aryl, C₁−C₂₀ heteroaryl, and C₂-C₂₀ heterocyclyl, each of which are substituted with one or more groups independently selected from H, F, Cl, Br, I, −CN, −NO₂, C₁−C₁₂ alkyl, C₂−C₁₂ alkenyl, C₂−C₁₂ alkynyl, C₁−C₁₂ heteroalkyl, −(C₁−C₁₂ heteroalkyldiyl)−(C₆−C₂₀ aryl), −(C₁−C₁₂ heteroalkyldiyl)−(C₆−C₂₀ aryldiyl)−(C₁−C₁₂ heteroalkyl), −(C₁−C₁₂ heteroalkyldiyl)−(C₆−C₂₀ aryldiyl)−(C₂-C₂₀ heterocyclyl), (C₁−C₆ alkyldiyl)−(C₆−C₂₀ aryl), −(C₁−C₆ alkyldiyl)−NR^aR^b, −(C₁−C₆ alkyldiyl)−OR^a, (C₁−C₆ alkyldiyl)−(C₃−C₂₀ carbocyclyl), (C₁-C₆ alkyldiyl)−(C₂-C₂₀ heterocyclyl), (C₁−C₆ alkyldiyl)−(C₁−C₂₀ heteroaryl), C₆−C₂₀ aryl, C₃−C₂₀ carbocyclyl, C₂−C₂₀ heterocyclyl, C₁−C₂₀ heteroaryl, −C(=NH)NH(OH), −C(=NH)NH₂, −C(=O)NR^aR^b, −C(=O)NR^a−NR^aR^b, −C(=O)NH(C₁-C₆ alkyldiyl)−NR^aR^b, −C(=O)OR^a, −NR^aR^b, −NR^aS(O)₂R^a, −NO2, −OR^a, −OC(=O)R^a, −SR^a, −S(O)R^a, −S(O)2R^a, −S(O)2NR^a, and −S(O)3H; R^a is independently selected from H, C1−C6 alkyl, phenyl, and benzyl, wherein phenyl and benzyl are optionally substituted with one or more groups independently selected from the group consisting of F, Cl, −CN, C₁−C₁₂ alkyl, C₂−C₁₂ alkenyl, and C₂−C₁₂ alkynyl; or where two R^a groups form a five-, six-, or seven-membered heteroaryl or heterocyclyl ring; R^b is independently selected from H, OH, C1−C6 alkyl, phenyl, and benzyl, wherein phenyl and benzyl are optionally substituted with one or more groups independently selected from the group consisting of F, Cl, −CN, C1−C12 alkyl, C2−C12 alkenyl, and C2−C12 alkynyl; R^c is selected from OH and −SO2F; R^d is selected from H and F; Cyc is a ring structure selected from the group consisting of C₃−C₂₀ carbocyclyl, C₆−C₂₀ aryl, C₂-C₂₀ heterocyclyl, C₁−C₂₀ heteroaryl, and −(C₁−C₂₀ heteroaryldiyl)−(C₁−C₁₂ heteroalkyl); n is 0 or 1; 371

15077.006WO1 R¹ is selected from the group consisting of H, C1−C12 alkyl, and C1−C12 heteroalkyl; R² is selected from the group consisting of H, C1−C12 alkyl, and C1−C12 heteroalkyl; or where R² forms a five- to ten-membered aryl, carbocyclyl, heteroaryl or heterocyclyl ring with A; X¹ is selected from the group consisting of H, −NHC(=O)−, (C₃−C₂₀ carbocyclyl)− C(=O)NH−, C₁−C₁₂ heteroalkyl, and C₁−C₂₀ heteroaryl; or where X¹ forms a five-, six-, or seven-membered heteroaryl or heterocyclyl ring with R¹; Sp is selected from the group consisting of a bond, O, NH, C₁−C₁₂ alkyldiyl, C₁−C₆₀ heteroalkyldiyl, C₃−C₂₀ carbocyclyldiyl, C₂-C₂₀ heterocyclyldiyl, C₆-C₂₀ aryldiyl, C₁−C₄₀ heteroaryldiyl, −(C₃−C₂₀ carbocyclyldiyl)−(C₁−C₆₀ heteroalkyldiyl)−, −(C₃−C₂₀ carbocyclyldiyl)−(C₁−C₁₂ alkyldiyl)−, −(C₂-C₂₀ heterocyclyldiyl)−(C₁−C₆₀ heteroalkyldiyl)−, −(C₂-C₂₀ heterocyclyldiyl)−(C₁−C₁₂ alkyldiyl)−, −(C₂-C₂₀ heterocyclyldiyl)−(C₂-C₂₀ heterocyclyldiyl)−, −(C₁−C₂₀ heteroaryldiyl)−(C₃−C₂₀ carbocyclyldiyl)−(C₁−C₆₀ heteroalkyldiyl)−, a solubilizing unit, and combinations thereof, where the solubilizing unit is selected from polyglycine, polysarcosine, polyethyleneoxy (PEG), a glycoside, C₁−C₆₀ heteroalkyldiyl, and combinations thereof; one of A, R^a, R^b, Cyc, R¹, R², and X¹ is attached to Sp; and each alkyl, alkyldiyl, alkenyl, alkenyldiyl, alkynyl, alkynyldiyl, heteroalkyl, heteroalkyldiyl, aryl, aryldiyl, carbocyclyl, carbocyclyldiyl, heterocyclyl, heterocyclyldiyl, heteroaryl, and heteroaryldiyl is independently and optionally substituted with one or more groups selected from F, Cl, Br, I, −CN, −CH3, −CH2CH3, −CH=CH2, −C^CH, −C^CCH3, − C^CCH₂NHCH₃, −CH₂CH₂CH₃, −CH(CH₃)₂, −CH₂CH(CH₃)₂, −CH₂OH, −CH₂OCH₃, − CH₂CH₂OH, −C(CH₃)₂OH, −CH(OH)CH(CH₃)₂, −C(CH₃)₂CH₂OH, −CH₂CH₂SO₂CH₃, − CH₂OP(O)(OH)₂, −CH₂F, −CHF₂, −CF₃, −CH₂CF₃, −CH₂CHF₂, −CH(CH₃)CN, −C(CH₃)₂CN, − CH₂CN, −CH₂NH₂, −CH₂NHSO₂CH₃, −CH₂NHCH₃, −CH₂N(CH₃)₂, −CO₂H, −COCH₃, − CO₂CH₃, −CO₂C(CH₃)₃, −COCH(OH)CH₃, −CONH₂, −CONHCH₃, −CON(CH₃)₂, − C(CH₃)₂CONH₂, −NH₂, −NHCH₃, −N(CH₃)₂, −NHCOCH₃, −N(CH₃)COCH₃, −NHS(O)₂CH₃, − N(CH₃)C(CH₃)₂CONH₂, −N(CH₃)CH₂CH₂S(O)₂CH₃, −NHC(=NH)H, −NHC(=NH)CH₃, − NHC(=NH)NH₂, −NHC(=O)NH₂, −NO₂, =O, −OH, −OCH₃, −OCH₂CH₃, −OCH₂CH₂OCH₃, − OCH2CH2OH, −OCH2CH2N(CH3)2, −OCH2F, −OCHF2, −OCF3, −OP(O)(OH)2, −S(O)2N(CH3)2, −SCH3, −S(O)2CH3, and −S(O)3H. 56. The compound of claim 55 wherein the VHL ligand has Formula IIb: 372

15077.006WO1 IIb. 57. The compound of claim 50 wherein the VHL ligand has Formula IIc: wherein: R³, R⁴, R⁵, and R⁶ are independently selected from the group consisting of H, F, Cl, Br, I, −CN, C1−C12 alkyl, C2−C12 alkenyl, C2−C12 alkynyl, C1−C12 heteroalkyl, −(C1−C12 heteroalkyldiyl)−(C6−C20 aryl), −(C1−C12 heteroalkyldiyl)−(C6−C20 aryldiyl)−(C1−C12 heteroalkyl), −(C1−C12 heteroalkyldiyl)−(C6−C20 aryldiyl)−(C2-C20 heterocyclyl), (C1−C6 alkyldiyl)−(C6−C20 aryl), −(C1−C6 alkyldiyl)−NR^aR^b, −(C1−C6 alkyldiyl)−OR^a, (C1−C6 alkyldiyl)−(C₃−C₂₀ carbocyclyl), (C₁-C₆ alkyldiyl)−(C₂-C₂₀ heterocyclyl), (C₁−C₆ alkyldiyl)−(C₁−C₂₀ heteroaryl), C₆−C₂₀ aryl, C₃−C₂₀ carbocyclyl, C₂−C₂₀ heterocyclyl, C₁−C₂₀ heteroaryl, −C(=NH)NH(OH), −C(=NH)NH₂, −C(=O)NR^aR^b, −C(=O)NR^a−NR^aR^b, −C(=O)NH(C₁-C₆ alkyldiyl)−NR^aR^b, −C(=O)OR^a, −NR^aR^b, −NO₂, −OR^a, −OC(=O)R^a, −SR^a, −S(O)R^a, −S(O)₂R^a, −S(O)₂NR^a, −S(O)₃H, and Sp; R^a is independently selected from H, C₁−C₆ alkyl, phenyl, and benzyl, wherein phenyl and benzyl are optionally substituted with one or more groups independently selected from the group consisting of F, Cl, −CN, C₁−C₁₂ alkyl, C₂−C₁₂ alkenyl, C₂−C₁₂ alkynyl, and Sp; or where two R^a groups form a five-, six-, or seven-membered heteroaryl or heterocyclyl group; 373

15077.006WO1 R^b is independently selected from H, OH, C1−C6 alkyl, phenyl, and benzyl, wherein phenyl and benzyl are optionally substituted with one or more groups independently selected from the group consisting of F, Cl, −CN, C₁−C₁₂ alkyl, C₂−C₁₂ alkenyl, C₂−C₁₂ alkynyl, and Sp; or where R³ and R⁴ together form a five-membered or six-membered heteroaryl or heterocyclyl group comprising one or more heteroatoms independently selected from N, O, P and S; X¹ is selected from the group consisting of H, and −NHCO−; or where X¹ forms a five-, six-, or seven-membered heteroaryl or heterocyclyl ring with R¹; one of X¹, R¹, R², R³, R⁴, R⁵, R⁶, R^a, R^b, and Cyc is attached to Sp; and each alkyl, alkyldiyl, alkenyl, alkenyldiyl, alkynyl, alkynyldiyl, heteroalkyl, heteroalkyldiyl, aryl, aryldiyl, carbocyclyl, carbocyclyldiyl, heterocyclyl, heterocyclyldiyl, heteroaryl, and heteroaryldiyl is independently and optionally substituted with one or more groups selected from F, Cl, Br, I, −CN, −CH₃, −CH₂CH₃, −CH=CH₂, −C^CH, −C^CCH₃, − CH₂CH₂CH₃, −CH(CH₃)₂, −CH₂CH(CH₃)₂, −CH₂OH, −CH₂OCH₃, −CH₂CH₂OH, −C(CH₃)₂OH, −CH(OH)CH(CH₃)₂, −C(CH₃)₂CH₂OH, −CH₂CH₂SO₂CH₃, −CH₂OP(O)(OH)₂, −CH₂F, −CHF₂, −CF₃, −CH₂CF₃, −CH₂CHF₂, −CH(CH₃)CN, −C(CH₃)₂CN, −CH₂CN, −CH₂NH₂, − CH₂NHSO₂CH₃, −CH₂NHCH₃, −CH₂N(CH₃)₂, −CO₂H, −COCH₃, −CO₂CH₃, −CO₂C(CH₃)₃, − COCH(OH)CH₃, −CONH₂, −CONHCH₃, −CON(CH₃)₂, −CONHS(O)₂N(CH₃)₂, − C(CH3)2CONH2, −NH2, −NHCH3, −N(CH3)2, −NHCOCH3, −N(CH3)COCH3, −NHS(O)2CH3, − N(CH3)C(CH3)2CONH2, −N(CH3)CH2CH2S(O)2CH3, −NHC(=NH)H, −NHC(=NH)CH3, − NHC(=NH)NH2, −NHC(=O)NH2, −NO2, =O, −OH, −OCH3, −OCH2CH3, −OCH2CH2OCH3, − OCH2CH2OH, −OCH2CH2N(CH3)2, −OCH2F, −OCHF2, −OCF3, −OP(O)(OH)2, −S(O)2N(CH3)2, −SCH3, −S(O)2CH3, and −S(O)3H. 58. A target protein binder and VHL ligand compound selected from Tables 1a-d. 59. A target protein binder and VHL ligand linker compound (TVL) having Formula III (TPI−Sp−VHL)−L³−Z III or a pharmaceutically acceptable salt, stereoisomer, mixture of stereoisomers, tautomer, or isotopic analog thereof, wherein: TPI is a target protein binder that binds to RIPK2, KRAS, Bcl-XL, or WEE1; VHL ligand has Formula IIIa: 374

15077.006WO1 or a pharmaceutically acceptable salt, stereoisomer, mixture of stereoisomers, tautomer, or isotopic analog thereof, wherein: A is a cyclic structure selected from C₃−C₂₀ carbocyclyl, C₆−C₂₀ aryl, C₁−C₂₀ heteroaryl, and C₂-C₂₀ heterocyclyl, each of which are substituted with one or more groups independently selected from H, F, Cl, Br, I, −CN, −NO₂, C₁−C₁₂ alkyl, C₂−C₁₂ alkenyl, C₂−C₁₂ alkynyl, C₁−C₁₂ heteroalkyl, −(C1−C12 heteroalkyldiyl)−(C6−C20 aryl), −(C1−C12 heteroalkyldiyl)−(C6−C20 aryldiyl)−(C₁−C₁₂ heteroalkyl), −(C₁−C₁₂ heteroalkyldiyl)−(C₆−C₂₀ aryldiyl)−(C₂-C₂₀ heterocyclyl), (C₁−C₆ alkyldiyl)−(C₆−C₂₀ aryl), −(C₁−C₆ alkyldiyl)−NR^aR^b, −(C₁−C₆ alkyldiyl)−OR^a, (C₁−C₆ alkyldiyl)−(C₃−C₂₀ carbocyclyl), (C₁-C₆ alkyldiyl)−(C₂-C₂₀ heterocyclyl), (C₁−C₆ alkyldiyl)−(C₁−C₂₀ heteroaryl), C₆−C₂₀ aryl, C₃−C₂₀ carbocyclyl, C₂−C₂₀ heterocyclyl, C₁−C₂₀ heteroaryl, −C(=NH)NH(OH), −C(=NH)NH₂, −C(=O)NR^aR^b, −C(=O)NR^a−NR^aR^b, −C(=O)NH(C₁-C₆ alkyldiyl)−NR^aR^b, −C(=O)OR^a, −NR^aR^b, −NR^aS(O)₂R^a, −NO₂, −OR^a, −OC(=O)R^a, −SR^a, −S(O)R^a, −S(O)₂R^a, −S(O)₂NR^a, and −S(O)₃H; R^a is independently selected from H, C₁−C₆ alkyl, phenyl, and benzyl, wherein phenyl and benzyl are optionally substituted with one or more groups independently selected from the group consisting of F, Cl, −CN, C₁−C₁₂ alkyl, C₂−C₁₂ alkenyl, and C₂−C₁₂ alkynyl; or where two R^a groups form a five-, six-, or seven-membered heteroaryl or heterocyclyl ring; R^b is independently selected from H, OH, C1−C6 alkyl, phenyl, and benzyl, wherein phenyl and benzyl are optionally substituted with one or more groups independently selected from the group consisting of F, Cl, −CN, C1−C12 alkyl, C2−C12 alkenyl, and C2−C12 alkynyl; R^c is selected from OH and −SO₂F; R^d is selected from H and F; 375

15077.006WO1 Cyc is a ring structure selected from the group consisting of C3−C20 carbocyclyl, C6−C20 aryl, C2-C20 heterocyclyl, C1−C20 heteroaryl, and −(C1−C20 heteroaryldiyl)−(C1−C12 heteroalkyl); n is 0 or 1; R¹ is selected from the group consisting of H, C₁−C₁₂ alkyl, and C₁−C₁₂ heteroalkyl; R² is selected from the group consisting of H, C₁−C₁₂ alkyl, and C₁−C₁₂ heteroalkyl; or where R² forms a five- to ten-membered aryl, carbocyclyl, heteroaryl or heterocyclyl ring with A; X¹ is selected from the group consisting of H, −NHC(=O)−, (C3−C20 carbocyclyl)− C(=O)NH−, C1−C12 heteroalkyl, and C1−C20 heteroaryl; or where X¹ forms a five-, six-, or seven-membered heteroaryl or heterocyclyl ring with R¹; Sp is selected from the group consisting of a bond, O, NH, C1−C12 alkyldiyl, C1−C60 heteroalkyldiyl, C3−C20 carbocyclyldiyl, C2-C20 heterocyclyldiyl, C6-C20 aryldiyl, C1−C40 heteroaryldiyl, −(C3−C20 carbocyclyldiyl)−(C1−C60 heteroalkyldiyl)−, −(C3−C20 carbocyclyldiyl)−(C1−C12 alkyldiyl)−, −(C2-C20 heterocyclyldiyl)−(C1−C60 heteroalkyldiyl)−, −(C2-C20 heterocyclyldiyl)−(C1−C12 alkyldiyl)−, −(C2-C20 heterocyclyldiyl)−(C2-C20 heterocyclyldiyl)−, −(C1−C20 heteroaryldiyl)−(C3−C20 carbocyclyldiyl)−(C1−C60 heteroalkyldiyl)−, a solubilizing unit, and combinations thereof, where the solubilizing unit is selected from polyglycine, polysarcosine, polyethyleneoxy (PEG), a glycoside, C1−C60 heteroalkyldiyl, and combinations thereof; TPI−Sp−VHL moiety is covalently attached to an antibody linker L³; Z is: where the wavy line is the attachment to L³; one of A, R^a, R^b, Cyc, R¹, R², and X¹ is attached to Sp; one of A, R^a, R^b, Cyc, R¹, R², X¹, and Sp is attached to L³; and each alkyl, alkyldiyl, alkenyl, alkenyldiyl, alkynyl, alkynyldiyl, heteroalkyl, heteroalkyldiyl, aryl, aryldiyl, carbocyclyl, carbocyclyldiyl, heterocyclyl, heterocyclyldiyl, heteroaryl, and heteroaryldiyl is independently and optionally substituted with one or more groups selected from F, Cl, Br, I, −CN, −CH₃, −CH₂CH₃, −CH=CH₂, −C^CH, −C^CCH₃, − C^CCH₂NHCH₃, −CH₂CH₂CH₃, −CH(CH₃)₂, −CH₂CH(CH₃)₂, −CH₂OH, −CH₂OCH₃, − 376

15077.006WO1 CH2CH2OH, −C(CH3)2OH, −CH(OH)CH(CH3)2, −C(CH3)2CH2OH, −CH2CH2SO2CH3, − CH2OP(O)(OH)2, −CH2F, −CHF2, −CF3, −CH2CF3, −CH2CHF2, −CH(CH3)CN, −C(CH3)2CN, − CH2CN, −CH2NH2, −CH2NHSO2CH3, −CH2NHCH3, −CH2N(CH3)2, −CO2H, −COCH3, − CO2CH3, −CO2C(CH3)3, −COCH(OH)CH3, −CONH2, −CONHCH3, −CON(CH3)2, − C(CH3)2CONH2, −NH2, −NHCH3, −N(CH3)2, −NHCOCH3, −N(CH3)COCH3, −NHS(O)2CH3, − N(CH3)C(CH3)2CONH2, −N(CH3)CH2CH2S(O)2CH3, −NHC(=NH)H, −NHC(=NH)CH3, − NHC(=NH)NH₂, −NHC(=O)NH₂, −NO₂, =O, −OH, −OCH₃, −OCH₂CH₃, −OCH₂CH₂OCH₃, − OCH₂CH₂OH, −OCH₂CH₂N(CH₃)₂, −OCH₂F, −OCHF₂, −OCF₃, −OP(O)(OH)₂, −S(O)₂N(CH₃)₂, −SCH₃, −S(O)₂CH₃, and −S(O)₃H. 60. The TVL compound of claim 59 wherein the TPI binds to RIPK2. 61. The TVL compound of claim 59 wherein the TPI binds to KRAS. 62. The TVL compound of claim 59 wherein the TPI binds to Bcl-XL. 63. The TVL compound of claim 59 wherein the TPI binds to WEE1. 64. The TVL compound of claim 59 wherein the VHL ligand has Formula IIIb: IIIb . 65. The TVL compound of claim 64 wherein the VHL ligand has Formula IIIc: IIIc 377

15077.006WO1 wherein: R³, R⁴, R⁵, and R⁶ are independently selected from the group consisting of H, F, Cl, Br, I, −CN, C₁−C₁₂ alkyl, C₂−C₁₂ alkenyl, C₂−C₁₂ alkynyl, C₁−C₁₂ heteroalkyl, −(C₁−C₁₂ heteroalkyldiyl)−(C₆−C₂₀ aryl), −(C₁−C₁₂ heteroalkyldiyl)−(C₆−C₂₀ aryldiyl)−(C₁−C₁₂ heteroalkyl), −(C₁−C₁₂ heteroalkyldiyl)−(C₆−C₂₀ aryldiyl)−(C₂-C₂₀ heterocyclyl), (C₁−C₆ alkyldiyl)−(C₆−C₂₀ aryl), −(C₁−C₆ alkyldiyl)−NR^aR^b, −(C₁−C₆ alkyldiyl)−OR^a, (C₁−C₆ alkyldiyl)−(C₃−C₂₀ carbocyclyl), (C₁-C₆ alkyldiyl)−(C₂-C₂₀ heterocyclyl), (C₁−C₆ alkyldiyl)−(C₁−C₂₀ heteroaryl), C₆−C₂₀ aryl, C₃−C₂₀ carbocyclyl, C₂−C₂₀ heterocyclyl, C₁−C₂₀ heteroaryl, −C(=NH)NH(OH), −C(=NH)NH2, −C(=O)NR^aR^b, −C(=O)NR^a−NR^aR^b, −C(=O)NH(C₁-C₆ alkyldiyl)−NR^aR^b, −C(=O)OR^a, −NR^aR^b, −NO₂, −OR^a, −OC(=O)R^a, −SR^a, −S(O)R^a, −S(O)2R^a, −S(O)2NR^a, and −S(O)3H; R^a is independently selected from H, C1−C6 alkyl, phenyl, and benzyl, wherein phenyl and benzyl are optionally substituted with one or more groups independently selected from the group consisting of F, Cl, −CN, C₁−C₁₂ alkyl, C₂−C₁₂ alkenyl, and C₂−C₁₂ alkynyl; or where two R^a groups form a five-, six-, or seven-membered heteroaryl or heterocyclyl group; R^b is independently selected from H, OH, C1−C6 alkyl, phenyl, and benzyl, wherein phenyl and benzyl are optionally substituted with one or more groups independently selected from the group consisting of F, Cl, −CN, C1−C12 alkyl, C2−C12 alkenyl, and C2−C12 alkynyl; or where R³ and R⁴ together form a five-membered or six-membered heteroaryl or heterocyclyl group comprising one or more heteroatoms independently selected from N, O, P and S; one of X¹, R¹, R², R³, R⁴, R⁵, R⁶, R^a, R^b, and Cyc is attached to Sp; one of X¹, R¹, R², R³, R⁴, R⁵, R⁶, R^a, R^b, and Cyc is attached to L³; and each alkyl, alkyldiyl, alkenyl, alkenyldiyl, alkynyl, alkynyldiyl, heteroalkyl, heteroalkyldiyl, aryl, aryldiyl, carbocyclyl, carbocyclyldiyl, heterocyclyl, heterocyclyldiyl, heteroaryl, and heteroaryldiyl is independently and optionally substituted with one or more groups selected from F, Cl, Br, I, −CN, −CH₃, −CH₂CH₃, −CH=CH₂, −C^CH, −C^CCH₃, − C^CCH₂NHCH₃, −CH₂CH₂CH₃, −CH(CH₃)₂, −CH₂CH(CH₃)₂, −CH₂OH, −CH₂OCH₃, − CH₂CH₂OH, −C(CH₃)₂OH, −CH(OH)CH(CH₃)₂, −C(CH₃)₂CH₂OH, −CH₂CH₂SO₂CH₃, − CH₂OP(O)(OH)₂, −CH₂F, −CHF₂, −CF₃, −CH₂CF₃, −CH₂CHF₂, −CH(CH₃)CN, −C(CH₃)₂CN, − CH2CN, −CH2NH2, −CH2NHSO2CH3, −CH2NHCH3, −CH2N(CH3)2, −CO2H, −COCH3, − CO2CH3, −CO2C(CH3)3, −COCH(OH)CH3, −CONH2, −CONHCH3, −CON(CH3)2, − CONHS(O)2N(CH3)2, −C(CH3)2CONH2, −NH2, −NHCH3, −N(CH3)2, −NHCOCH3, − 378

15077.006WO1 N(CH3)COCH3, −NHS(O)2CH3, −N(CH3)C(CH3)2CONH2, −N(CH3)CH2CH2S(O)2CH3, − NHC(=NH)H, −NHC(=NH)CH3, −NHC(=NH)NH2, −NHC(=O)NH2, −NO2, =O, −OH, −OCH3, −OCH2CH3, −OCH2CH2OCH3, −OCH2CH2OH, −OCH2CH2N(CH3)2, −OCH2F, −OCHF2, − OCF3, −OP(O)(OH)2, −S(O)2N(CH3)2, −SCH3, −S(O)2CH3, and −S(O)3H. 66. The TVL compound of claim 59 wherein L³ has the formula: −Str¹−(PEP)_y−(IM)_z− wherein: Str¹ is a stretcher unit covalently attached to Z; PEP is a protease-cleavable, peptide unit covalently attached to Str¹ and IM or the TPI- VHL moiety when y is 1; IM is an immolative unit covalently attached the TPI-VHL moiety when z is 1; y is 0 or 1; and z is 0 or 1. 67. The TVL compound of claim 59 wherein TPI has the formula: 379

15077.006WO1 380

15077.006WO1 where the wavy line indicates the point of attachment to the spacer unit Sp. 68. A TVL compound selected from Tables 2a-d. 381

15077.006WO1 69. A degrader antibody conjugate composition prepared by conjugation of a cysteine amino acid of an antibody with a TVL compound of claim 59. 70. A degrader antibody conjugate composition prepared by conjugation of a cysteine amino acid of an antibody with a TVL compound selected from Tables 2a-d. 71. A degrader antibody conjugate composition prepared by conjugation of a cysteine amino acid of an antibody with a TVL compound of claim 59. 72. A process for preparing a degrader antibody conjugate composition comprising reacting a cysteine amino acid of an antibody with a TVL compound of claim 59 whereby a degrader antibody conjugate composition is formed. 73. A pharmaceutical composition comprising a therapeutically effective amount of the degrader antibody conjugate composition of any one of claims 1 to 54, and one or more pharmaceutically acceptable diluent, vehicle, carrier or excipient. 74. A method for treating cancer comprising administering a therapeutically effective amount of the pharmaceutical composition of claim 73, to a patient in need thereof, wherein the cancer is selected from hematological and solid tumors, wherein the cancer is selected from breast cancer, triple negative breast cancer, lung cancer, non-small cell lung cancer, small cell lung cancer, colorectal cancer, and lymphoma, diffuse large B-cell lymphoma (DLBCL), follicular cancer, leukemia, and acute myelogenous leukemia (AML). 75. The method of claim 74 wherein the degrader antibody conjugate of the pharmaceutical composition modulates the level of a target protein in the patient selected from RIPK2, KRAS, Bcl-XL, and WEE1. 76. Use of a degrader antibody conjugate composition of any one of claims 1 to 54 in the manufacture of a medicament for the treatment of cancer in a mammal. 77. A degrader antibody conjugate composition of any one of claims 1 to 54 for use in a method for treating cancer. 382