Compounds and methods for targeted protein degradation

Abstract

A compound including formula (I) or formula (II) for use in promoting targeted protein degradation.

Description

PATENT COMPOUNDS AND METHODS FOR TARGETED PROTEIN DEGRADATION RELATED APPLICATION

[0001] This application claims priority from U. S. Provisional ApplicationNo. 63 / 735,365, filed December 18, 2024, the subject matter of which is incorporated herein by reference in its entirety.BACKGROUND

[0002] Cell maintenance and normal function requires controlled degradation of cellular proteins. For example, degradation of regulatory proteins triggers events in the cell cycle, such as DNA replication, chromosome segregation, etc. Accordingly, such degradation of proteins has implications for the cell's proliferation, differentiation, and death. While inhibitors of proteins can block or reduce protein activity in a cell, protein degradation is another possibility to reduce activity or remove the target protein completely. Utilizing a cell's protein degradation pathway can, therefore, provide a means for reducing or removing protein activity. One of the cells major degradation pathways is known as the ubiquitin-proteasome system. In this system, a protein is marked for proteasomal degradation by an E3 ubiquitin ligase that binds to the protein and transfers ubiquitin molecules to the protein. The E3 ubiquitin ligase is part of a pathway that includes El and E2 ubiquitin ligases, which make ubiquitin available to the E3 ubiquitin ligase catalyzed transfer to the protein.

[0003] TRIM21 is an E3 ubiquitin ligase that has been well studied in immunology, where it is known to oligomerize and ubiquitinate immunoglobulin G (IgG)-coated pathogens to mediate their rapid degradation. The ability of TRIM21 to degrade antibodybound cargoes has also been harnessed for the TRIM-AWAY platform, in which cellular delivery of an antibody against a target protein can induce its TRIM21 -mediated degradation. Recent reports have also explored the potential of this approach as a gene therapy, for example by delivering viruses encoding TRIM21 -nanobody fusion proteins that target aggregation-prone proteins like Tau.SUMMARY

[0004] Embodiments described herein relate to compounds and methods for targeted protein degradation (TPD), and particularly to E3 ubiquitin ligase tripartite motif 21 (TRIM21) targeted probes, molecular glue degraders, and bifunctional or proteolysistargeting chimeric compounds (PROTACs) for TPD. TPD utilizes molecular glues or PROTACs to eliminate disease-causing proteins by promoting their interaction with E3 ubiquitin ligases.

[0005] We identified PRLX-93936 and BMS-214662 as molecular glues that directly target the E3 ligase, TRIM21, to induce degradation of nucleoporin proteins, leading to inhibition of nuclear export and ultimately cell death. Eoss of nucleoporins and nuclear export accounts for past observations in which BMS-214662 led to disrupted subcellular protein localization. Furthermore, the cytotoxicity of these compounds correlates with TRIM21 expression across hundreds of cancer cell lines, suggesting clinical re-evaluation of these agents in patients with TRIM21-high cancers.

[0006] Relative to recently-reported TRIM21 -targeting glues, these compounds and analogs based on these compounds display high cellular potency, creating new opportunities for targeted protein degradation via the design of additional glues and the design of PRO TACs. Functionalization of PRLX-93936 into PRO TACs enabled selective degradation of multimeric proteins, such as those within biomolecular condensates, while sparing monomeric proteins. Such PROTACs can find utility as modulators of targeted ubiquitination of a variety of polypeptides and other proteins, which are then degraded and / or otherwise inhibited by the PROTACs described herein. An advantage of the compounds described herein is that a broad range of pharmacological activities is possible, consistent with the degradation / inhibition of targeted polypeptides from virtually any protein class or family. As aberrant protein assemblies cause diseases, such as autoimmunity, neurodegeneration, and cancer, our findings highlight the potential of TRIM21 -based multimer-selective degraders as a strategy to treat the direct causes of these diseases.

[0007] Advantageously, TRIM21 is the ideal E3 ligase for the development of protein aggregate PROTACs, especially in the context of tau aggregates. “Substrate clustering activation’’ of TRIM21 is what potentiates its unique competency in the development of aggregate-specific TPD relative to other E3 ligases. TRIM21’s E3 ligase activity is first activated and then enhanced when several TRIM21 homodimers associate via their RING domains at a protein target. Unlike E3 ligases, such as CRBN and VHL, non-intermolecular associated TRIM21 does not have E3 ligase activity until further TRIM21 dimers are recruited to a target. This can be leveraged for aggregate selective targeting of TPD pathological tau aggregates.

[0008] In some embodiments, a TRIM21 binding moiety (or ligand for TRIM21) or compound that is used for TPD can have the structure of formula (I):R4(I) or a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinR1is absent, halogen, alkyl, haloalkyl, or alkoxy;R2aand R2bare each independently -H, halogen, alkyl, or haloalkyl, or alternatively R2aand R2btogether form a cycloalkyl or heterocyclyl;R3and R5are each independently -H, halogen, alkyl, haloalkyl, or oxo;R4is absent, -H, halogen, alkyl, haloalkyl, or -C(O)-alkyl optionally substituted with one or more halogen;R6and R7are each independently -H, halogen, alkyl, or haloalkyl;R8is absent, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, -alkylene-OH optionally substituted with one or more halogen, -CN, -C(O)-N(R10)2, -C(O)O-alkyl optionally substituted with one or more halogen, or carboxyl;R9is -H, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, or -O-alkylene-alkynyl optionally substituted with one or more halogen;each R10is independently -H, alkyl, or haloalkyl;X is N or O; oralternatively, one of R1, R2a, R2b, R3, R4, R5, R6, R8, or R9is substituted with a linker that is optionally linked to a target protein ligand; andwherein at least one of R1, R2a, R2b, R3, R4, R5, R6, R7, or R8is not absent or -H if X is N and R9is alkoxy, such as ethoxy.

[0009] In some embodiments, R1is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, or Ci-C6alkoxy.

[0010] In some embodiments, R1is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF, methoxy, or ethoxy.

[0011] In some embodiments, R2aand R2bare each independently -H, halogen Ci-Ce alkyl, or Ci-Ce haloalkyl, or R2aand R2btogether form a C3-C6 cycloalkyl.

[0012] In other embodiments, one of R2aor R2bis -H, -F, -Cl, -Br, -I, -CF3, methyl, ethyl, or isopropyl, and the other of R2aor R2bis -H, or R2aand R2btogether form cyclopropyl.

[0013] In some embodiments, R3is -H, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, or oxo.

[0014] In other embodiments, R is -H, -F, -CF3, methyl, ethyl, isopropyl, or oxo.

[0015] In some embodiments, X is N and R4is -H, Ci-Ce alkyl, or -C(O)-(Ci-Ce alkyl).

[0016] In other embodiments, X is N and R4is -H, methyl, ethyl, isopropyl, -C(O)mcthyl, -C(O)ethyl, or -C(O)isopropyl.

[0017] In still other embodiments, X is O and R4is absent.

[0018] In some embodiments, R3is -H, halogen, Ci-Ce alkyl, or Ci-Ce haloalkyl.

[0019] In other embodiments, R5is -H, -F, -Cl, -Br, -I, -CF3, methyl, ethyl, or isopropyl.

[0020] In some embodiments, R6is -H, halogen, Ci-Ce alkyl, or Ci-Ce haloalkyl.

[0021] In other embodiments, R6is -H, -F, -Cl, -Br, -I, -CF3, methyl, ethyl, or isopropyl.

[0022] In some embodiments, R7is -H, halogen, Ci-Ce alkyl, or Ci-Ce haloalkyl.

[0023] In some embodiments, R7is -II, -F, -Cl, -Br, -I, -CF3, methyl, ethyl, or isopropyl.

[0024] In some embodiments, R8is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, Ci-Ce alkoxy, -(Ci-Cealkylene)-OH, -CN, -C(O)-N(H)(Ci-Ce alkyl), -C(O)O-(Ci-Ce alkyl), or carboxyl.

[0025] In other embodiments, R8is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, ethoxy, propoxy, -methylene-OH, ethylene-OH, -propylene- OH, -CN, -C(O)-N(H)(methyl), -C(O)-N(H)(ethyl), -C(O)-N(H)(propyl), -C(O)O-(methyl), -C(O)O-(ethyl), -C(O)O-(propyl), or carboxyl.

[0026] In some embodiments, R9is -II, halogen, Ci-Ce alkyl, Ci-Ce alkoxy, or -O-(Ci-Ce alkylene)-alkynyl.

[0027] In other embodiments, R9is -H, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF methoxy, ethoxy, propoxy, or -O-(methylene)-ethynyl.

[0028] In some embodiments, one of R1, R2a, R2b, R3, R4, R5, R6, R8, or R9is substituted with a linker that is linked to the target protein ligand.

[0029] In some embodiments, the linker includes an aliphatic linker, a cyclic linker or a polyethylene glycol linker.

[0030] In some embodiments, a compound of formula (I) does not have the structure of:or a pharmaceutically acceptable salt, tautomer, or solvate thereof.

[0031] In other embodiments, R1is absent, halogen, alkyl, haloalkyl, or alkoxy; R2ais alkyl or haloalkyl; R2bis -II; R3, R5, R6, and R7are each independently -II, halogen, alkyl, or haloalkyl; R4is -H; R8is absent, halogen, alkyl, haloalkyl, -alkylene-OH, -CN, -C(O)-N(R10)2, or -C(O)O-alkyl; R9is alkoxy or -O-alkylene-alkynyl; each R10is independently -H or alkyl; and X is N; or alternatively, one of R1, R2a, R2b, R3, R4, R5, R6, R8, or R9is substituted with a linker that is optionally linked to a target protein ligand.

[0032] In other embodiments, R1is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, Ci-Ce alkoxy; R2ais Ci-Ce alkyl or Ci-Ce haloalkyl; R2bis -H; R3, R5, R6, and R7are each independently -H, halogen, Ci-Ce alkyl, or Ci-Ce haloalkyl; R4is -H; R8is absent, halogen, Ci-C6alkyl, Ci-C6haloalkyl, Ci-C6alkoxy, -(Ci-C6alkylene)-OH, -CN, -C(O)-N(H)(CI-C6alkyl), -C(O)O-(Ci-Ce alkyl), or carboxyl; and R9is Ci-Ce alkoxy or -O-(Ci-Ce alkylene)-alkynyl; and X is N; or alternatively, one of R1, R2a, R2b, R3, R4, R5, R6, R8, or R9is substituted with a linker that is optionally linked to a target protein ligand.

[0033] In other embodiments, R1is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl,-CF3, methoxy, or ethoxy; R2ais methyl, ethyl, isopropyl, or -CF3; R2bis -H; R3, R?. R6, and R7are each independently -H, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, or -CF3; R4is -H; R8is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, ethoxy, propoxy, -methylene-OH, ethylene-OH, -propylene-OH, -CN, -C(O)-N(H)(methyl), -C(O)-N(H)(ethyl), -C(O)-N(H)(propyl), -C(O)O-(methyl), -C(O)O-(ethyl), -C(O)O-(propyl), or carboxyl; R9is methoxy, ethoxy, propoxy, or -O-(methylene)-ethynyl; and X is N; or alternatively, one of R1, R2a, R2b, R3, R4, R5, R6, R8, or R9is substituted with a linker that is optionally linked to a target protein ligand.

[0034] In some embodiment, the compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof comprises an atropisomer of the compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof, wherein the atropisomer is substantially free of its opposite enantiomer or pharmaceutically acceptable salt, tautomer, or solvate thereof.

[0035] In other embodiments, a TRIM21 binding moiety (or ligand for TRIM21) or compound that is used for TPD can have the structure of formula (IA):R4(IA) or a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinR1is absent, halogen, alkyl, haloalkyl, or alkoxy;R2ais -H, halogen, alkyl, or haloalkyl;R3is -H, halogen, alkyl, or haloalkyl;R4is -H, halogen, alkyl, haloalkyl, or -C(O)-alkyl optionally substituted with one or more halogen;R8is absent, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, -alkylene-OH optionally substituted with one or more halogen, -CN, -C(O)-N(R10)2, -C(O)O-alkyl optionally substituted with one or more halogen, or carboxyl;each R10is independently -H, alkyl, or haloalkyl; oralternatively, one of R1, R2a, R3, R4, or R8is substituted with a linker that is optionally linked to a target protein ligand; andwherein at least one of R1, R2a, R3, R4, or R8is not absent or -H.

[0036] In some embodiments, R1is absent, halogen, Ci-C& alkyl, Ci-Ce haloalkyl, or Ci-C6alkoxy.

[0037] In other embodiments, R1is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, or ethoxy.

[0038] In some embodiments, R2ais -H, halogen, Ci-Ce alkyl, or Ci-Ce haloalkyl.

[0039] In other embodiments, R2ais -H, -F, -CF3, methyl, ethyl, or isopropyl.

[0040] In some embodiments, R3is -H, halogen, Ci-Ce alkyl, or Ci-Ce haloalkyl.

[0041] In other embodiments, R3is -H, -F, -CF3, methyl, ethyl, or isopropyl.

[0042] In some embodiments, R4is -H, Ci-Ce alkyl, or -C(O)-(Ci-Ce alkyl).

[0043] In other embodiments, R4is -H, methyl, ethyl, isopropyl, -C(O)methyl, -C(O)ethyl, or -C(O)isopropyl.

[0044] In some embodiments, R8is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, Ci-Ce alkoxy, -(Ci-C6alkylene)-OH, -CN, -C(O)-N(H)(CI-C6alkyl), -C(O)O-(Ci-C6alkyl), or carboxyl.

[0045] In other embodiments, R8is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, ethoxy, propoxy, -methylene-OH, ethylene-OH, -propylene- OH, -CN, -C(O)-N(H)(methyl), -C(O)-N(H)(ethyl), -C(O)-N(H)(propyl), -C(O)O-(methyl), -C(O)O-(ethyl), -C(O)O-(propyl), or carboxyl.

[0046] In some embodiments, the compound of formula (I A) or pharmaceutically acceptable salt, tautomer, or solvate thereof can include an atropisomer of the compound of formula (IA) or pharmaceutically acceptable salt, tautomer, or solvate thereof, wherein the atropisomer is substantially free of its opposite enantiomer or pharmaceutically acceptable salt, tautomer, or solvate thereof.

[0047] In some embodiments, the atropisomer is a P isomer.

[0048] In other embodiments, a TRIM21 binding moiety (or ligand for TRIM 21) or compound that is used for TPD can have the structure of formula (IB):(IB) or a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinR1is absent, halogen, alkyl, haloalkyl, or alkoxy;R2ais alkyl or haloalkyl;R3is -H, halogen, alkyl, or haloalkyl;R4is -H, halogen, alkyl, haloalkyl, or -C(O)-alkyl optionally substituted with one or more halogen;R8is absent, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, -alkylene-OH optionally substituted with one or more halogen, -CN, -C(O)-N(R10)2, -C(O)O-alkyl optionally substituted with one or more halogen, or carboxyl;each R10is independently -H, alkyl, or haloalkyl; oralternatively, one of R1, R2a, R3, R4, or R8is substituted with a linker that is optionally linked to a target protein ligand.

[0049] In some embodiments, R1is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, or Ci-C6alkoxy.

[0050] In other embodiments, R1is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, or ethoxy.

[0051] In some embodiments, R2ais Ci-Ce alkyl or Ci-Ce haloalkyl.

[0052] In other embodiments, R2ais -CF3, methyl, ethyl, or isopropyl.

[0053] In some embodiments, R3is -H, halogen, Ci-Ce alkyl, or Ci-Ce haloalkyl.

[0054] In other embodiments, R3is -H, -F, -CF3, methyl, ethyl, or isopropyl.

[0055] In some embodiments, R4is -H, Ci-Ce alkyl, or -C(O)-(Ci-C6 alkyl).

[0056] In other embodiments, R4is -H, methyl, ethyl, isopropyl, -C(O)methyl, -C(O)ethyl, or -C(O)isopropyl.

[0057] In some embodiments, R8is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, Ci-C6alkoxy, -(Ci-C6alkylene)-OH, -CN, -C(O)-N(H)(CI-C6alkyl), -C(O)O-(Ci-C6alkyl), or carboxyl.

[0058] In other embodiments, R8is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, ethoxy, propoxy, -methylene-OH, ethylene-OH, -propylene-OH, -CN, -C(O)-N(H)(methyl), -C(O)-N(H)(ethyl), -C(O)-N(H)(propyl). -C(O)O-(methyl), -C(O)O-(ethyl), -C(O)O-(propyl), or carboxyl.

[0059] In some embodiments, the compound of formula (IA) or pharmaceutically acceptable salt, tautomer, or solvate thereof can include an atropisomer of the compound of formula (IA) or pharmaceutically acceptable salt, tautomer, or solvate thereof, wherein the atropisomer is substantially free of its opposite enantiomer or pharmaceutically acceptable salt, tautomer, or solvate thereof.

[0060] In some embodiments, the atropisomer is a P isomer.

[0061] In other embodiments, a TRIM21 binding moiety (or ligand for TRIM21) or compound that is used for TPD can include an atropisomer of formula (IC):(IC) or a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinR1is absent, halogen, alkyl, haloalkyl, or alkoxy;R2ais alkyl or haloalkyl;R8is absent, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, -alkylene-OH optionally substituted with one or more halogen, -CN, -C(O)-N(R10)2, -C(O)O-alkyl optionally substituted with one or more halogen, or carboxyl;each R10is independently -H, alkyl, or haloalkyl; oralternatively, one of R1, R2a, or R8is substituted with a linker that is optionally linked to a target protein ligand; andwherein the compound of formula (IC) or pharmaceutically acceptable salt, tautomer, or solvate thereof is substantially free of its opposite enantiomer or pharmaceutically acceptable salt, tautomer, or solvate thereof.

[0062] In some embodiments, R1is absent, halogen, Ci-Ce alkyl, Ci-C.6 haloalkyl, or C1-C6 alkoxy.

[0063] In other embodiments, R1is absent, -F. -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, or ethoxy.

[0064] In some embodiments, R2ais C1-C6 alkyl or Ci-Ce haloalkyl.

[0065] In other embodiments, R2ais -CF3, methyl, ethyl, or isopropyl.

[0066] In some embodiments, R8is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, Ci- C6alkoxy, -(Ci-C6alkylene)-OH, -CN, -C(O)-N(H)(CI-C6alkyl), -C(O)O-(Ci-C6alkyl), or carboxyl.

[0067] In other embodiments, R8is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, ethoxy, propoxy, -methylene-OH, ethylene-OH, -propylene-OH, -CN, -C(O)-N(H)(methyl), -C(O)-N(H)(ethyl), -C(O)-N(H)(propyl), -C(O)O-(methyl), -C(O)O-(ethyl), -C(O)O-(propyl), or carboxyl.

[0068] In some embodiments, the atropisomer is a P isomer.

[0069] In some embodiments, the compound of formula (IC) or pharmaceutically acceptable salt, tautomer, or solvate thereof has a shorter retention time or elutes faster by chiral separation than its opposite enantiomer or pharmaceutically acceptable salt, tautomer, or solvate thereof.

[0070] In other embodiments, a TRIM21 binding moiety (or ligand for TRIM 21) or compound that is used for TPD can have the structure of formula (II):(II) or a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinR11is -H, alkyl, or haloalkyl;R12and R13are each independently absent, -CN, alkyl, haloalkyl, haloalkenyl, alkynyl, or -alkylene-alkynyl optionally substituted with one or more halogen;R14is cycloalkyl, hctcrocyclyl, aryl, or hctcroaryl, each of which is optionally substituted with R17;R15and R16are each independently absent, halogen, alkyl, or haloalkyl;R17is halogen, alkyl, haloalkyl, or alkoxy;R18is -H, alkyl, or haloalkyl;X1is S(O)n, CH2, or CO;n is 0, 1, or 2;alternatively, one of R11, R12, R13, R14, R13, R16, R17, or R18is substituted with a linker that is optionally linked to a target protein ligand; and wherein at least one of R12and / or R13is not -CN if X1is S(O)2and R14is a thiophenyl group.

[0071] In some embodiments, R11is H, Ci-Ce alkyl, or Ci-Ce haloalkyl.

[0072] In other embodiments, R11is -H, methyl, or ethyl, preferably -H.

[0073] In some embodiments, R12and R13are each independently absent, -CN, Ci-Ce alkyl, Ci-Ce haloalkyl, C2-Ce haloalkenyl, C2-C<> alkynyl, -(Ci-Ce alkylene)-(C2-Ce alkynyl).

[0074] In other embodiments, R12and R13are each independently absent, -CN, or ethynyl.

[0075] In some embodiments, R14is C3-C.8 cycloalkyl, heterocyclyl having 5 to 10 ring atoms, C6-C10 aryl, or heteroaryl having 5 to 10 ring atoms, each of which is optionally substituted with R17.

[0076] In other embodiments, R14is thiophenyl, phenyl, isoxazolyl, or thiazolyl, each of which is optionally substituted with R17.

[0077] In some embodiments, R15and R16are each independently absent, halogen, Ci- C6alkyl, or Ci-C6haloalkyl.

[0078] In some embodiments, R17is halogen, Ci-Ce alkyl, or Ci-Ce alkoxy.

[0079] In some embodiments, X1is S(O)2.

[0080] In some embodiments, one of R11, R12, R13, R14, R15, R16, R17, or R18is substituted with a linker that is linked to the target protein ligand.

[0081] In some embodiments, the linker includes an aliphatic linker, a cyclic linker, or a polyethylene glycol linker.

[0082] In some embodiments, a PROTAC that includes a compound of formula (I) or a pharmaceutically acceptable salt, tautomer, or solvate thereof linked to a target protein ligand can have the structure of:R4(I) or a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinR1is absent, halogen, alkyl, haloalkyl, or alkoxy;R2aand R2bare each independently -H, halogen, alkyl, or haloalkyl, or alternatively R2aand R2btogether form a cycloalkyl or heterocyclyl;R3and R5are each independently -H, halogen, alkyl, haloalkyl, or oxo;R4is absent -H, halogen, alkyl, haloalkyl, or -C(O)-alkyl optionally substituted with one or more halogen;R6and R7are each independently -H, halogen, alkyl, or haloalkyl;R8is absent, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, -alkylene-OH optionally substituted with one or more halogen, -CN, -C(O)-N(R10)2, -C(O)O-alkyl optionally substituted with one or more halogen, or carboxyl;R9is -H, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, or -O-alkylene-alkynyl optionally substituted with one or more halogen;each R10is independently -H, alkyl, or haloalkyl;X is N or O; andwherein one of R1, R2a, R2b, R3, R4, R5, R6, R8, R9is substituted with a linker; anda target protein ligand is linked to the linker.

[0083] In some embodiments, R1is absent, halogen, Ci-C& alkyl, Ci-Ce haloalkyl, or Ci-C6alkoxy.

[0084] In some embodiments, R2aand R2bare each independently -H, halogen Ci-Ce alkyl, or Ci-Ce haloalkyl, or R2aand R2btogether foim a C3-C6 cycloalkyl.

[0085] In some embodiments, R3is -H, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, or oxo.

[0086] In some embodiments, X is N and R4is -H, Ci-Ce alkyl, or -C(O)-(Ci-Ce alkyl).

[0087] In some embodiments, R3is -H, halogen, Ci-Ce alkyl, or Ci-Ce haloalkyl.

[0088] In some embodiments, R6is -H, halogen, Ci-Ce alkyl, or Ci-Ce haloalkyl.

[0089] In some embodiments, R7is -H, halogen, Ci-Ce alkyl, or Ci-Ce haloalkyl.

[0090] In some embodiments, R8is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, Ci-Ce alkoxy, -(Ci-Ce alkylene)-OH, -CN, -C(O)-N(H)(Ci-Ce alkyl), -C(O)O-(Ci-C6 alkyl), or carboxyl.

[0091] In some embodiments, R9is -H, Ci-Ce alkyl, Ci-Ce alkoxy, or -O-(Ci-Ce alkylene)-alkynyl.

[0092] In other embodiments, a PROTAC that includes a compound of formula (I) or a pharmaceutically acceptable salt, tautomer, or solvate thereof linked to a target protein ligand can include the formula selected from:or a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinR1is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, or Ci-Ce alkoxy;R2ais Ci-C6alkyl or Ci-C6haloalkyl;R8is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, Ci-Ce alkoxy, -(Ci-Ce alkylene)-OH, -CN, -C(O)-N(H)(Ci-Ce alkyl), -C(O)O-(Ci-Ce alkyl), or carboxyl;R9is -H, Ci-C,6 alkyl, Ci-Ce alkoxy, or -O-(Ci-Ce alkylene)-alkynyl;L is the linker; andTPL is the target protein ligand.

[0093] In still other embodiments, a PROTAC that includes a compound of formula (I) or a pharmaceutically acceptable salt, tautomer, or solvate thereof linked to a target protein ligand can include the formula selected from:or a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinR1is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, or Ci-Ce alkoxy;R2ais Ci-C6alkyl or Ci-C6haloalkyl;R8is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, Ci-Ce alkoxy, -(Ci-Ce alkylene)-OH, -CN, -C(O)-N(H)(Ci-Ce alkyl), -C(O)O-(Ci-Ce alkyl), or carboxyl;L is the linker; andTPL is the target protein ligand.

[0094] In some embodiments, the PROTAC including the compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof is an atropisomer or pharmaceutically acceptable salt, tautomer, or solvate thereof that is substantially free of its opposite enantiomer or pharmaceutically acceptable salt, tautomer, or solvate thereof.

[0095] In some embodiments, the atropisomer is a P isomer.

[0096] In some embodiments, the linker includes an aliphatic linker, a cyclic linker, or a polyethylene glycol linker.

[0097] In other embodiments, a PROTAC that includes a compound of formula (II) or a pharmaceutically acceptable salt, tautomer, or solvate thereof linked to target protein ligand can have the structure of:(II) or a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinR11is H, alkyl, or haloalkyl;R12and R13are each independently absent, -CN, alkyl, haloalkyl, haloalkenyl, alkynyl, or -alkylene-alkynyl optionally substituted with one or more halogen;R14is cycloalkyl, heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with R17;R15and R16are each independently absent, halogen, alkyl, or haloalkyl;R17is halogen, alkyl, or alkoxy;R18is H, alkyl, or haloalkyl;X1is S(O)n, CH2, or CO;n is 0, 1, or 2;wherein one R11, R12, R13, R14, R15. R16, R17, or R18is substituted with a linker, and a target protein ligand is linked to the linker.

[0098] In some embodiments, R11is H, Ci-Ce alkyl, Ci-Cc, haloalky 1, preferably -H.

[0099] In some embodiments, R12and R13are each independently absent, -CN, Ci-Ce alkyl, Ci-Cehaloalkyl, C2-Cehaloalkenyl, C2-C6 alkynyl, -(Ci-Ce alkylene)-(C2-Ce alkynyl).

[0100] In some embodiments, R14is Ca-Cs cycloalkyl, heterocyclyl having 5 to 10 ring atoms, Ce-Cio aryl, or heteroaryl having 5 to 10 ring atoms, each of which is optionally substituted with R17preferably R14is thiophenyl, phenyl, isoxazolyl, or thiazolyl, each of which is optionally substituted with R17.

[0101] In some embodiments, X1is S(O)2.

[0102] In some embodiments, R15and R16are each independently absent, halogen, Ci- C6 alkyl, or C1-C6 haloalkyl.

[0103] In some embodiments, R17is halogen, Ci-Ce alkyl, or Ci-Ce alkoxy.

[0104] In some embodiments, a PROTAC that includes a compound of formula (II) or a pharmaceutically acceptable salt, tautomer, or solvate thereof linked to a target protein ligand can include the formula selected from:pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinR14is Ca-Cs cycloalkyl, heterocyclyl having 5 to 10 ring atoms, Ce-Cio aryl, or heteroaryl having 5 to 10 ring atoms, each of which is optionally substituted with R17;R15and R16are each independently absent, halogen, C1-C6 alkyl, or C1-C6 haloalkyl;R17is halogen, alkyl, or alkoxy;L1is the linker; andTPL is the target protein ligand.

[0105] In other embodiments, a PROTAC that includes a compound of formula (II) or a pharmaceutically acceptable salt, tautomer, or solvate thereof linked to a target protein ligand can include the formula selected from:; or a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinL1is the linker; andTPL is the target protein ligand.

[0106] In some embodiments, the proteins that can be targeted by the target protein ligand can include B7.1 and B7, TINFRlm, TNFR2, NADPH oxidase, BclIBax and other partners in the apoptosis pathway, C5a receptor, HMG-CoA reductase, PDE V phosphodiesterase type, PDE IV phosphodiesterase type 4, PDE I, PDEII, PDEIII, squalene cyclase inhibitor, CXCR1, CXCR2, nitric oxide (NO) synthase, cyclo-oxygenase 1, cyclooxygenase 2, alpha- sy nuclein, HIV capsid protein, VP40, BET-bromodomain protein, condensate-forming oncogenic fusion proteins including nucleoporin-containing fusion proteins (e.g., NUP98-NSD1, NUP98-KDM5A, NUP98-PHF23; DEK-NUP214, NUP214-ABL1, SET-NUP214, etc.), EML4-ALK, and BRD4-NUT, or oligomeric proteins involved in inflammatory signaling including NLRP3, RIPK3, RIPK1, MLKL, and GSDM3, 5HT receptors, dopamine receptors, G Proteins, Gq, histamine receptors, 5-lipoxygenase, tryptase serine protease, thymidylate synthase, purine nucleoside phosphorylase, GAPDH trypanosomal, glycogen phosphorylase, Carbonic anhydrase, chemokine receptors, JAW STAT, RXR and similar, HIV 1 protease, HIV 1 integrase, influenza, neuramimidase, hepatitis B reverse transcriptase, sodium channel, multi drug resistance (MDR), protein P-glycoprotein (and MRP), tyrosine kinases, CD23, CD124, tyrosine kinase p56 lek, CD4, CD5, IL-2 receptor, IL-1 receptor, TNF-aR, ICAM1, CaH- channels, VCAM, VLA-4 integrin, selectins, CD40 / CD40L, newokinins and receptors, inosine monophosphate dehydrogenase, p38 MAP Kinase, Ras / Raf / ME / ERK pathway, interleukin- 1 converting enzyme, caspase, HCV, NS3 protease, HCV NS3 RNA helicase, glycinamide ribonucleotide formyl transferase, rhinovirus 3C protease, herpes simplex virus- 1 (HSV-I), protease, cytomegalovirus (CMV) protease, poly (ADP-ribose) polymerase, cyclin dependent kinases, vascular endothelial growth factor, c-Kit, TGFP activated kinase I, mammalian target of rapamycin, SHP2, androgen receptor, oxytocin receptor, microsomal transfer protein inhibitor, bile acid transport inhibitor, 5 alpha reductase inhibitors, angiotensin 11, glycine receptor, noradrenaline reuptake receptor, estrogen receptor, estrogen related receptors, focal adhesion kinase, Src, endothelin receptors, neuropeptide Y and receptor, adenosine receptors, adenosine kinase and AMP deaminase, purinergic receptors (P2Y1, P2Y2, P2Y4, P2Y6, P2X1-7), famesyltransferases, geranylgeranyl transferase, TrkA a receptor for NGF, amyloid, tau or pathological tau aggregates, tyrosine kinase Flk-IIKDR, vitronectin receptor,integrin receptor, Her-21 neu, telomerase inhibition, cytosolic phospholipaseA2, or EGF receptor tyrosine kinase.

[0107] In still other embodiments, the proteins that can be targeted by the target protein ligand include tau, amyloid, alpha-synuclein, HIV capsid protein, VP40, condensate-forming oncogenic fusion proteins including nucleoporin-containing fusion proteins (e.g., NUP98-NSD1, NUP98-KDM5A, NUP98-PHF23; DEK-NUP214, NUP214-ABE1, SET-NIJP214, etc.) EML4-ALK, and BRD4-NUT, or oligomeric proteins involved in inflammatory signaling including NLRP3, RIPK3, RIPKl, MLKL, and GSDM3.

[0108] In some embodiments, the PROTAC promotes targeted protein degradation, such as pathological protein degradation or pathological aggregated protein degradation.

[0109] In some embodiments, the PROTAC can be used in targeted protein degradation, such as pathological protein degradation or pathological aggregated protein degradation.

[0110] In some embodiments, the PROTAC can be used for targeted degradation of pathogenic tau.

[0111] In some embodiments, the PROTAC can be used for aggregated protein degradation and treatment of central nervous system diseases, neurodegenerative diseases, and other diseases, such as Alzheimer’s disease and frontotemporal dementia.

[0112] Other embodiments relate to a method of modulating tau protein, its ubiquitination, and the subsequent degradation in a subject, e.g., a cell, a tissue, mammal, or human patient. The method can include administering an effective amount of a PROTAC that binds to tau as described herein to a subject in need thereof. The PROTAC can be effective in modulating tau ubiquitination and degradation in the subject.

[0113] Still other embodiments relate to a method of treating or ameliorating a symptom of a disease related to tau activity in a subject, e.g., a cell, a tissue, mammal, or human patient. The method can include administering an effective amount of a PROTAC that binds to tau as described herein to a subject in need thereof. The PROTAC can be effective in treating or ameliorating a symptom of a disease related to tau activity in the subject. In certain embodiments, the disease to be treated is a neurological or a neurodegenerative disease, e.g., Alzheimer's, Parkinson's, Dementia, etc.

[0114] In other embodiments, the PROTAC can be used in a method of treating cancer.BRIEF DESCRIPTION OF THE DRAWINGS

[0115] Figs. l(A-H) illustrate schematics, plots, and structures showing a synthetic rescue screen identifies PRLX-93936 as inducing cell death via a ubiquitin-dependent mechanism. A) Schematic of the synthetic rescue screening strategy. Desired molecules show reduction of cytotoxicity when El Ubiquitin Activating Enzyme activity is impaired by the UBA1 inhibitor TAK-243, indicative of a mechanism of cell death reliant on ubiquitination. Created in BioRender. Adams, D. (2025). B) Dot-plot representing the performance of 1754 bioactive small molecules in the synthetic rescue screen performed using OCI- AML-3 cells treated for 10 h. x-axis, viability measured for each library molecule with only DMSO vehicle treatment; y-axis, viability measured for each library molecule cotreated with TAK-243 (200 nM). Hits (red oval) were significantly cytotoxic as single agents (viability <50%, x-axis) and showed at least 30% greater viability in combination with TAK-243. Red circle, PRLX-93936. Each point represents data from a single well. C) Structures of PRLX-93936 and crastin. D) Cell viability (CcllTitcr-Glo) following treatment with PRLX-93936 alone or in combination with TAK-243 (200 nM) or Bortezomib (500 nM) for 10 h in OCI- AML-3 cells (n = 2 independent experiments, each with at least 2 wells per concentration). E, F) Pearson correlation plots establishing that TRIM21 expression (log2(Transcripts per million+1) is correlated with PRLX-93936 cytotoxicity (log2(Fold Change) (E) and that the cytotoxicity of PRLX-93936 and BMS-214662 is uniquely well correlated with TRIM21 among the small molecules within the database (F). Pearson correlation p-values were derived from two-sided tests with no correction for multiple comparisons. Top 1000 correlates are shown. Each correlation represents data from at least 453 cell lines. G) Structures of BMS-214662 and BMS-225975. II) Cell viability (CellTiter-Glo) following treatment with PRLX-93936 alone or in combination with TAK-243 (200 nM) or Bortezomib (500 nM) for 10 h in OCI-AML-3 cells (n = 2 independent experiments, each with at least 2 wells per concentration). Source data are provided as Source Data files.

[0116] Figs. 2(A-H) illustrate a western blot and plots showing TRIM21 loss prevents cell killing by PRLX-93936 and BMS-214662. A, B) Western blot following CRLSPR / Cas9 targeting of TRIM21 in OCI-AML-3 cells (A) and JURKAT cells (B). Representative of n = 1 independent experiments. C, D) Cell viability following treatment with PRLX-93936 for 24 h in OCI-AML-3 cells (c) or JURKAT cells (D). E, F) Cell viability measurements asdescribed for c, d but using BMS-214662. G) Cell viability following treatment with erastin for 72 h in OCI- AML-3 cells. H) Cell viability following treatment with BMS-225975 for 72 h in JURKAT cells. All cell viability measurements used CellTiter-Glo. All CTG data represents n =2 independent experiments, each with al least 2 wells per condition, except h which is n = 1 with 2 wells per condition. Source data are provided as Source Data Files.

[0117] Figs. 3(A-L) illustrate western blots and plots showing overexpression of TRIM21 drives sensitivity to PRLX-93936 and BMS-214662, which directly target TRIM21. A, D, G Western blot following lenti viral overexpression of TRIM21-FLAG or inactive mutant TRIM21CA-FLAG in OCI-AML-3 cells (A), C33A cells (D), or HEK293T cells (G). Representative of n = 1 independent experiment. B, E, H Cell viability following treatment with PRLX-93936 for 24 h in TRIM21 -expressing OCI-AML-3 cells (B) or 72 h in TRIM21-cxprcssing C33A (E) or HEK293T cells (H) (n = 2 independent experiments, each with at least 2 wells per concentration, except (h), which shows two replicate wells from n = 1 independent experiment). C, F, I Cell viability following treatment with BMS-214662 for 24 h in TRIM21-FLAG-expressing OCI-AML-3 (C), C33A cells (F), or HEK293T cells (I), n =2 independent experiments, each with at least 3 wells per concentration, except (I) which shows two independent replicate wells from one experiment. J, K CETSA experiment monitoring the impact of PRLX-93936 (j; n = 1) or BMS-214662 (k; n =2 independent experiments) on the thermal denaturation of TRIM21-FLAG in OCI-AML-3 cells.L Immunoprecipitation assay in which TRIM21 is captured by an IgG-coated resin in the presence of PRLX-93936 analog 1 (Fig. 11 and Fig. 6A for characterization of this analog) at indicated concentrations. IP = Immunoprecipitation, FT = Flow-through. IP and FT / Input samples from the same experiment were run on separate gels in parallel. Representative of n = 1 independent experiments. Source data are provided as Source Data Files.

[0118] Figs. 4(A-M) illustrate western blots, plots, and images showing PRLX-93936 and BMS-214662 induce degradation of nucleoporin protein and impair nuclear transport. A Label-free LC / MSMS quantitation (LFQ) of protein levels following PRLX-93936 treatment (500 nM, 6 h) in Jurkat cells (x-axis) and OCI-AML-3 sgScramble (sgSCR) cells (y-axis), with nucleoporin proteins high-lighted in orange. B, C Volcano plot highlighting alterations in protein levels by LFQ following treatment with PRLX-93936 in OCI-AML-3 sgSCR cells (B) and OCI-AML-3 sgTRIM21 cells (C) with nucleoporin proteins labeled in blue. FC = fold change. D, E LFQ intensity values for specific nucleoporins noted panels a-c in bothsgSCR and sgTRIM21 OCI-AML-3 cells. F Heatmap showing fold change in nucleoporin abundance by LFQ after treatment of OCI-AML-3 cells with PRLX-93936 (500 nM) for 1, 2, or 4 h. Represents mean of n = 3 independent replicates. G Volcano plot as in b except OCI-AML-3 sgSCR cells are treated with BMS-214662 (1 p. 4 h). H, I LFQ intensity values for specific nucleoporins in both sgSCR and sgTRIM21 OCI-AML-3 cells following treatment with BMS-214662 (1 pM, 4 h). Scatter and volcano plot points represent mean of n = 3 independent replicates. Volcano plot P- values derived from two-sided t tests, no correction for multiple comparisons. Bar graph points represent one of n - 3 independent replicates. J Western blot demonstrating reduction of NUP214 and NUP88 protein levels with increasing concentrations of PRLX-93936 treatment (4 h, OCI-AML-3 cells). K Western blot evaluating the impact of pre-treatment (2 h) with the proteasome inhibitor bortczomib (5 pM) or UB Al inhibitor TAK-243 (0.5 pM) prior to addition of PRLX-93936 (1 pM, 4 h). Western blot results in (J, K) representative of n = 1 independent experiments. 1 Quantitation of immunofluorescence imaging of RANBP1 reported as the ratio of nuclear to cytoplasmic signal intensity in the indicated C33A overexpression cell lines. LMB = Leptomycin B; PRLX = PRLX-93936; BMS = BMS-214662. All treatments 6 h. Points represent average of 2 wells / condition with >900 cells quantified / condition from n = 2 independent experiments, m Representative images of RANBP1 subcellular localization following the treatments in 1 (n = 2 independent experiments). Panels (A, B) and (G, H) include data from the same experiment. Source data are provided as Source Data Files.

[0119] Figs. 5(A-II) illustrate plots and western blots showing PRLX-93936 and BMS-214662 phenotypes can be abrogated by disrupting the NUP98 autoproteolysis domain. A) Structure of hydroxy-acepromazine. B, C) Viability (CellTiter-Glo) of OCI-AML-3 cells (WT or TRIM21 KO) following 24 h treatment with the indicated concentrations of hydroxy-acepromazine (B) or PRLX-93936 (C). D F) A549 cells were subjected to CRISPR / Cas9 targeting using 5 sgRNAs targeting the autoproteolysis domain of NUP98 and then exposed to PRLX-93936 (1 pM) for 7 days. Surviving cells were then exposed to the indicated concentrations of PRLX-93936 (D), BMS-214662 (E) or TAK-243 (F) and cell viability was measured after 72 h with CellTiter-Glo. All CellTiter-Glo experiments represent n = 2 independent experiments with at least 2 wells per condition. G) Western blot of NUP98 in WT A549 as well as A549 cells targeted with the five independent NUP98 sgRNAs. A NUP98 proteoform (-200 kDa) indicative of impaired autoproteolysis is highlighted. H)Western blot of NUP214 and NUP88 following PRLX-93936 treatment (1 pM / 12 h) in A549 cells resistant to PRLX-93936 following treatment with four of the NUP98 sgRNAs reported in D-G. Western blotting results in (G, H) representative of n = 1 independent experiments. Source data are provided as Source Data Files.

[0120] Figs. 6(A-H) illustrate plots, structures, and images showing identification of PRLX-93936 analogs with enhanced potency and a PRLX-93936-derived TRTMTAC. A-C) Viability (CellTiter-Glo) of OCI-AML-3 cells following 24 h treatment with the indicated concentrations of the drawn PRLX-93936 analogs, n - 3 independent biological replicates are shown, each representing the mean of two independent wells. D) Representative images of an A549 PML-eGFP-BRD4(BD2) clone expressing TRIM21-FLAG treated with indicated compounds for 8 h. 31-35 each 10 pM; see Fig. 9D for structures of 31-33. Representative of n = 1 independent experiments. E) Quantification of EGFP foci in A549 PML-cGFP-BRD4(BD2) clone expressing TRIM21-FLAG treated with indicated compounds in dose for 8 h. Points indicate two independent wells derived from n = 1 independent experiment with >50 quantified cells per dose. F) Representative images as in (D). 34, 10 pM; HGClg, 10 pM; BTZ, Bortezomib, 2 pM. Representative of n = 2 independent experiments, g Image quantification as in (e) for the experiment described in (F). N = 2 independent experiments are shown, each point represented as the mean of two independent wells and >100 quantified cells per dose. H) Structures of 34 and 35. Absolute configuration of the chiral axis of 34 / 35 is unknown, and the assignment is arbitrary. Source data are provided as Source Data Files.

[0121] Figs. 7 (A-C) illustrate plots showing cell viability (CellTiter-Glo) following treatment of OCI-AML-3 cells with TAK-243 at the indicated concentrations for either 10 h (blue) or 24 h (red). These data informed selection of 20 nM TAK-243 for our rescue screen in Fig. IB. n=l independent experiment with 2 wells / condition. B) Cell viability (CellTiter-Glo) for the four additional hits that proved to be false positives and did not show suppression of cytotoxicity upon retest in OCI-AML-3 cells. n=2 independent experiment with points representing mean of 2 wells / condition. C) Pearson correlation of PRLX-93936 cytotoxicity expressed as log2fold change (PRISM) and TRIM21 transcript level expressed as log2(Transcripts per million+1) across the cancer cell lines available within DepMap. n=859 cell lines. A cytotoxicity value of 0 indicates no effect, while more negative numbers reflect more potent cytotoxic effects. Source data are provided as Source Data files.

[0122] Figs. 8(A-H) illustrate plots, structures, and immunoprecipitation assays showing: A) Cell viability measurements of OCI-AML-3 treated with PRLX-93936 (10 pM) for 24 hours. Bars represent mean of n=3 independent experiments and error bars represent standard deviation. P = 0.0080 (Welch’s / -lest, two-sided). B) Cell viability measurements of WT or TRIM21 KO OCI-AML-3 or JURKAT cells treated with DMSO for 24 hours. n=2 independent runs are shown. C) Cell viability measurements of OCI-AML-3 or C33A cells expressing EGFP or TRIM21 and treated with DMSO for 24 hrs. Points represent mean of 16 wells from n=2 independent experiments. D) Quantification of Western Blot from Fig. 3J by Densitometry showing thermal stabilization of cellular TRIM21-FLAG by PRLX-93936. FLAG bands normalized to Actin then normalized to ice-treated control. Representative of n=l independent experiment. E) Quantification of Western Blot from Fig. 3K by Densitometry showing thermal stabilization of cellular TRIM21-FLAG by BMS-214662. Normalization as in Fig. 8D. Representative of n=2 independent experiments. F) Quantification of Western Blot from Fig. 31 by Densitometry as in Fig. 8D. Representative of n=l independent experiments. G) Immunoprecipitation assay in which recombinant 6x-His TRIM21 PRY-SPRY captured on IgG resin is displaced in the presence of BMS-214662. Representative of n=l independent experiments. H) Viability of OCI-AML-3 cells (CellTiter-Glo) treated with the indicated concentrations of PRLX-93936 (left) or 1 (right) plus the indicated concentrations of established TRIM21 ligand HGClg. (n=2 independent experiments, each with at least 3 wells per concentration). Source data are provided as Source Data files.

[0123] Figs. 9(A-H) illustrate plots and images showing further proteomic characterization of PRLX-93936 and BMS-214662. A) Label-free LC / MSMS quantitation (LFQ) of protein levels following PRLX-93936 treatment (500 nM, 6 hr) in JURKAT cells (x-axis) and OCI-AML-3 cells (y-axis), with alterations in nucleoporin proteins highlighted in orange. This expanded version of Fig. 4A shows all 4 quadrants. Each point represents the mean of n=3 independent replicates. B) Volcano plot highlighting alterations in protein levels following treatment with PRLX-93936 (500 nM, 6 hr) in JURKAT cells. FC = fold change. Nucleoporin proteins colored in blue. C, D) LFQ intensity values for specific nucleoporins in both sgSCR (c) and sgTRIM21 (D) OCI-AML-3 cells. E) Individual LFQ values for the n=3 independent samples averaged to produce the heatmap shown in Fig. 4F. F) Volcano plot as in b highlighting alterations in protein levels following treatment withBMS-214662 (1 pM, 4 hr) in OCI- AML-3 TRIM21 KO cells. G) LFQ intensity values for specific nucleoporins in both OCI-AML-3 cells treated with BMS-214662 (black squares) or BMS-225975 (1 pM, 4 hr), the N-methylated analog of BMS-214662 that lacks TRIM21-dependent cell killing (open circles). Scatter and volcano plot points represent mean of n=3 independent replicates. Volcano plot P-values derived from two-sided t-tests, no correction for multiple comparisons. Bar graph points represent one of n=3 independent replicates. H) Representative immunofluorescence images for RANBP1 as in Fig. 4M, but now false colored with RANBP1 red and DAPI blue. Source data are provided as Source Data files.

[0124] Figs. 10(A-G) illustrate A) analytical chiral SFC traces for 9 and 10, the separated atropisomers of PRLX-93936, following 4 d at RT in ethanol. No racemization is observed. B) Viability (CellTiter-Glo) of OCI-AML-3 cells following 24 h treatment with the indicated concentrations of the drawn PRLX-93936 analogs. Mean of N=3 independent biological replicates are shown, individual biological replicates shown in Fig. 11.C) Viability (CellTiter-Glo) of OCI-AML-3 cells following 24 hours of treatment with the indicated concentrations of the enantiomeric PRLX-93936 analogs 11 and 12. N=2 independent replicates are shown. D) Structures of all candidate PROTACs assessed.Absolute configuration of the chiral axis of 34 / 35 is unknown, and the assignment is arbitrary. E) Representative colored images from Fig 6d of an A549 PML-EGFP-BRD4(BD2) clone expressing TRIM21-FLAG treated with indicated compounds for 8 hrs.31-35 each 10 pM. Nuclei are marked with Hoechst (DNA). Representative of n=l independent experiments. Scale bar (white) = 20 pm. F) Quantification of EGFP foci in an A549 PML-EGFP-BRD4(BD2) clone expressing TRIM21-FLAG pre-treated with indicated compounds for 2 hrs before treatment with indicated concentrations of 31, 32, 33, or 35 for 8 hrs. Experiments representative of n=2 (35) or n=l (31, 32, 33) independent experiments, each containing 2 independent wells and >100 quantified cells per dose. G) Representative colored images from Figure 4F as in (E). 34, 10 pM; HGClg, 10 pM; BTZ, Bortezomib, 2 pM. Representative of n=2 independent experiments and >100 quantified cells per dose. Scale bar (white) = 20 pm. Source data are provided as Source Data files.

[0125] Fig. 11 illustrates a table showing PRLX-93936 analogs ability to induce TRIM21 -mediated cell death in both OCI-AML-3 and JURKAT cells.DETAILED DESCRIPTION

[0126] While the following terms are believed to be well understood by one of ordinary skill in the art, the following definitions are set forth to facilitate explanation of the presently disclosed subject matter.

[0127] As used herein, the verb “comprise” as is used in this description and in the claims and its conjugations are used in its non-limiting sense to mean that items following the word are included, but items not specifically mentioned are not excluded. The present invention may suitably “comprise”, “consist of’, or “consist essentially of”, the steps, elements, and / or reagents described in the claims.

[0128] It is further noted that the claims may be drafted to exclude any optional element. As such, this statement is intended to serve as antecedent basis for use of such exclusive terminology as "solely", "only" and the like in connection with the recitation of claim elements, or the use of a "negative" limitation.

[0129] The term “pharmaceutically acceptable” means suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit / risk ratio, and effective for their intended use within the scope of sound medical judgment.

[0130] The term “pharmaceutically acceptable salts” include those obtained by reacting the active compound functioning as a base, with an inorganic or organic acid to form a salt, for example, salts of hydrochloric acid, sulfuric acid, phosphoric acid, methanesulfonic acid, camphorsulfonic acid, oxalic acid, maleic acid, succinic acid, citric acid, formic acid, hydrobromic acid, benzoic acid, tartaric acid, fumaric acid, salicylic acid, mandelic acid, carbonic acid, etc. Those skilled in the art will further recognize that acid addition salts may be prepared by reaction of the compounds with the appropriate inorganic or organic acid via any of a number of known methods. The term “pharmaceutically acceptable salts” also includes those obtained by reacting the active compound functioning as an acid, with an inorganic or organic base to form a salt, for example salts of ethylenediamine, N-methyl-glucamine, lysine, arginine, ornithine, choline, N, N'-dibenzylethylenediamine, chloroprocaine, diethanolamine, procaine, N-benzylphenethylamine, diethylamine, piperazine, tris-(hydroxymethyl)-aminomethane, tetramethylammonium hydroxide, triethylamine, dibenzylamine, ephenamine, dehydroabietylamine, N-ethylpiperidine,benzylamine, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, ethylamine, basic amino acids, and the like. Non limiting examples of inorganic or metal salts include lithium, sodium, calcium, potassium, magnesium salts and the like.

[0131] Additionally, the salts of the compounds described herein, can exist in either hydrated or unhydrated (the anhydrous) form or as solvates with other solvent molecules. Non-limiting examples of hydrates include monohydrates, dihydrates, etc. Nonlimiting examples of solvates include ethanol solvates, acetone solvates, etc.

[0132] The term "solvates" means solvent addition forms that contain either stoichiometric or non- stoichiometric amounts of solvent. Some compounds have a tendency to trap a fixed molar ratio of solvent molecules in the crystalline solid state, thus forming a solvate. If the solvent is water the solvate formed is a hydrate, when the solvent is alcohol, the solvate formed is an alcoholate. Hydrates are formed by the combination of one or more molecules of water with one of the substances in which the water retains its molecular state as H2O, such combination being able to form one or more hydrate.

[0133] 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 superimposable on their mirror image partner.

[0134] 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.

[0135] The term "enantiomers" as used herein, refers to two stereoisomers of a compound.

[0136] The term "atropisomers" refers to conformational stereoisomers which occur when rotation about a single bond in the molecule is prevented, or greatly slowed, as a result of steric interactions with other parts of the molecule and the substituents at both ends of the single bond are asymmetrical, i.e., they do not require a stereocenter. Where the rotational barrier about the single bond is high enough, and interconversion between conformations is slow enough, separation and isolation of the isomeric species may be permitted.Atropisomers are enantiomers without a single asymmetric atom.

[0137] The compounds and salts described herein can exist in several tautomeric forms, including the enol and imine form, and the keto and enamine form and geometric isomers and mixtures thereof. Tautomers exist as mixtures of a tautomeric set in solution. In solidform, usually one tautomer predominates. Even though one tautomer may be described, the present application includes all tautomers of the present compounds. A tautomer is one of two or more structural isomers that exist in equilibrium and are readily converted from one isomeric form to another. This reaction results in the formal migration of a hydrogen atom accompanied by a switch of adjacent conjugated double bonds. In solutions where tautomerization is possible, a chemical equilibrium of the tautomers will be reached. The exact ratio of the tautomers depends on several factors, including temperature, solvent, and pH. The concept of tautomers that are interconvertable by tautomerizations is called tautomerism.

[0138] Of the various types of tautomerism that are possible, two are commonly observed. In keto-enol tautomerism a simultaneous shift of electrons and a hydrogen atom occurs.

[0139] Tautomerizations can be catalyzed by: Base: 1. deprotonation; 2. formation of a delocalized anion (e.g., an enolate); 3. protonation at a different position of the anion; Acid: 1. protonation; 2. formation of a delocalized cation; 3. deprotonation at a different position adjacent to the cation.

[0140] The terms below, as used herein, have the following meanings, unless indicated otherwise:“Amino” refers to the -NH2 radical.“Cyano” refers to the -CN radical.“Ilalo” or “halogen” refers to bromo, chloro, fluoro or iodo radical.“Hydroxy” or “hydroxyl” refers to the -OH radical.“Imino” refers to the =NH substituent.“Nitro” refers to the -NO2 radical.“Oxo” refers to the =0 substituent.“Thioxo” refers to the =S substituent.

[0141] “Alkyl” or “alkyl group” refers to a fully saturated, straight or branched hydrocarbon chain radical having from one to twelve carbon atoms, and which is attached to the rest of the molecule by a single bond. Alkyls comprising any number of carbon atoms from 1 to 12 are included. An alkyl comprising up to 12 carbon atoms is a C1-C12 alkyl, an alkyl comprising up to 10 carbon atoms is a C1-C10 alkyl, an alkyl comprising up to 6 carbon atoms is a Ci-Ce alkyl and an alkyl comprising up to 5 carbon atoms is a C1-C5 alkyl. A Ci-C.5 alkyl includes Cs alkyls, C4 alkyls, C3 alkyls, C2 alkyls and Ci alkyl (z.e., methyl). A Ci-Ce alkyl includes all moieties described above for C1-C5 alkyls but also includes G, alkyls. A C1-C10 alkyl includes all moieties described above for C1-C5 alkyls and Ci-Ce alkyls, but also includes C7, Cs, C9 and C10 alkyls. Similarly, a C1-C12 alkyl includes all the foregoing moieties, but also includes C11 and C12 alkyls. Non-limiting examples of C1-C12 alkyl include methyl, ethyl, n-propyl, i-propyl, sec-propyl, n-butyl, i-butyl, sec-butyl, t-butyl, n-pentyl, t-amyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl, and n-dodecyl.Unless stated otherwise specifically in the specification, an alkyl group can be optionally substituted.

[0142] “Alkylene” or “alkylene chain” refers to a fully saturated, straight or branched divalent hydrocarbon chain radical, and having from one to twelve carbon atoms. Nonlimiting examples of C1-C12 alkylene include methylene, ethylene, propylene, n-butylcnc, ethenylene, propenylene, n-butenylene, propynylene, zz-butynylene, and the like. The alkylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond. The points of attachment of the alkylene chain to the rest of the molecule and to the radical group can be through one carbon or any two carbons within the chain. Unless stated otherwise specifically in the specification, an alkylene chain can be optionally substituted.

[0143] “Alkenyl” or “alkenyl group” refers to a straight or branched hydrocarbon chain radical having from two to twelve carbon atoms, and having one or more carbon-carbon double bonds. Each alkenyl group is attached to the rest of the molecule by a single bond. Alkenyl group comprising any number of carbon atoms from 2 to 12 are included. An alkenyl group comprising up to 12 carbon atoms is a C2-C12 alkenyl, an alkenyl comprising up to 10 carbon atoms is a C2-C10 alkenyl, an alkenyl group comprising up to 6 carbon atoms is a C2-C6 alkenyl and an alkenyl comprising up to 5 carbon atoms is a C2-C5 alkenyl. A C2-C5 alkenyl includes C5 alkenyls, C4 alkenyls, C3 alkenyls, and C2 alkenyls. A C2-C6 alkenyl includes all moieties described above for C2-C5 alkenyls but also includes Ce alkenyls. A C2-C10 alkenyl includes all moieties described above for C2-C5 alkenyls and C2-C6 alkenyls, but also includes C7, Cs, C9 and C10 alkenyls. Similarly, a C2-C12 alkenyl includes all the foregoing moieties, but also includes Cn and C12 alkenyls. Non-limiting examples of C2-C12 alkenyl include ethenyl (vinyl), 1 -propenyl, 2-propenyl (allyl), iso-propenyl, 2-methyl-l-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 1-heptenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl, 5-heptenyl, 6-heptenyl, 1-octenyl, 2-octenyl, 3-octenyl, 4-octenyl, 5-octenyl, 6-octenyl, 7 -octenyl, 1-nonenyl, 2-nonenyl, 3-nonenyl, 4-nonenyl, 5-nonenyl, 6-nonenyl, 7-nonenyl, 8-nonenyl, 1 -decenyl, 2-decenyl, 3-decenyl, 4-decenyl, 5-decenyl, 6-decenyl, 7-decenyl, 8-decenyl, 9-decenyl, 1 -undecenyl, 2-undecenyl, 3-undecenyl, 4-undecenyl, 5-undecenyl, 6-undecenyl, 7-undecenyl, 8-undecenyl, 9-undecenyl, 10-undecenyl, 1-dodecenyl, 2-dodecenyl, 3-dodecenyl, 4-dodecenyl, 5-dodecenyl, 6-dodecenyl, 7-dodecenyl, 8-dodecenyl, 9-dodecenyl, 10-dodecenyl, and 11 -dodecenyl. Unless stated otherwise specifically in the specification, an alkyl group can be optionally substituted.

[0144] “Alkenylene” or “alkenylene chain” refers to a straight or branched divalent hydrocarbon chain radical, having from two to twelve carbon atoms, and having one or more carbon-carbon double bonds. Non-limiting examples of C2-C12 alkcnylcnc include ethene, propene, butene, and the like. The alkenylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond. The points of attachment of the alkenylene chain to the rest of the molecule and to the radical group can be through one carbon or any two carbons within the chain. Unless stated otherwise specifically in the specification, an alkenylene chain can be optionally substituted.

[0145] “Alkynyl” or “alkynyl group” refers to a straight or branched hydrocarbon chain radical having from two to twelve carbon atoms, and having one or more carbon-carbon triple bonds. Each alkynyl group is attached to the rest of the molecule by a single bond. Alkynyl group comprising any number of carbon atoms from 2 to 12 are included. An alkynyl group comprising up to 12 carbon atoms is a C2-C12 alkynyl, an alkynyl comprising up to 10 carbon atoms is a C2-C10 alkynyl, an alkynyl group comprising up to 6 carbon atoms is a C2-C6 alkynyl and an alkynyl comprising up to 5 carbon atoms is a C2-C5 alkynyl. A C2-C5 alkynyl includes C5 alkynyls, C4 alkynyls, C3 alkynyls, and C2 alkynyls. A C2-C6 alkynyl includes all moieties described above for C2-C5 alkynyls but also includes (7> alkynyls. A C2-C10 alkynyl includes all moieties described above for C2-C5 alkynyls and C2-C6 alkynyls, but also includes C7, Cs, C9 and C10 alkynyls. Similarly, a C2-C12 alkynyl includes all the foregoing moieties, but also includes Cn and C12 alkynyls. Non-limiting examples of C2-C12 alkenyl include ethynyl, propynyl, butynyl, pentynyl and the like. Unless stated otherwise specifically in the specification, an alkyl group can be optionally substituted.

[0146] “Alkynylene” or “alkynylene chain” refers to a straight or branched divalent hydrocarbon chain radical, having from two to twelve carbon atoms, and having one or more carbon-carbon triple bonds. Non-limiting examples of C2-C12 alkynylene include ethynylene, propargylene and the like. The alkynylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond. The points of attachment of the alkynylene chain to the rest of the molecule and to the radical group can be through one carbon or any two carbons within the chain. Unless stated otherwise specifically in the specification, an alkynylene chain can be optionally substituted.

[0147] “Alkoxy” refers to a radical of the formula -ORawhere Rais an alkyl, alkenyl or alknyl radical as defined above containing one to twelve carbon atoms. Unless stated otherwise specifically in the specification, an alkoxy group can be optionally substituted.

[0148] “Alkylcarbonyl” refers to the -C(=O)Ramoiety, wherein Rais an alkyl, alkenyl or alkynyl radical as defined above. A non-limiting example of an alkyl carbonyl is the methyl carbonyl (“acetal”) moiety. Alkylcarbonyl groups can also be referred to as “Cw-Czacyl” where w and z depicts the range of the number of carbon in Ra, as defined above. For example, “C1-C10 acyl” refers to alkylcarbonyl group as defined above, where Rais C1-C10 alkyl. C2-C10 alkenyl, or C2-C10 alkynyl radical as defined above. Unless stated otherwise specifically in the specification, an alkyl carbonyl group can be optionally substituted.

[0149] “Aryl” refers to a hydrocarbon ring system radical comprising hydrogen, 6 to 18 carbon atoms and at least one aromatic ring. For purposes of this invention, the aryl radical can be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which can include fused or bridged ring systems. Aryl radicals include, but are not limited to, aryl radicals derived from phenyl (benzene), aceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, chrysene, fluoranthene, fluorene, fl.v-indacene,.v-indacene, indane, indene, naphthalene, phenalene, phenanthrene, pleiadene, pyrene, and triphenylene. Unless stated otherwise specifically in the specification, the term “aryl” is meant to include aryl radicals that are optionally substituted.

[0150] “Aralkyl” or “arylalkyl” refers to a radical of the formula -Rb-Rcwhere Rb is an alkylene group as defined above and Rc is one or more aryl radicals as defined above.Aralkyl radicals include, but are not limited to, benzyl, diphenylmethyl and the like. Unless stated otherwise specifically in the specification, an aralkyl group can be optionally substituted.

[0151] “Carbocyclyl,” “carbocyclic ring” or “carbocycle” refers to a ring structure, wherein the atoms which form the ring are each carbon. Carbocyclic rings can comprise from 3 to 20 carbon atoms in the ring. Carbocyclic rings include aryls and cycloalkyl. Cycloalkenyl and cycloalkynyl as defined herein. Unless stated otherwise specifically in the specification, a carbocyclyl group can be optionally substituted.

[0152] “Cycloalkyl” refers to a stable non-aromatic monocyclic or polycyclic fully saturated hydrocarbon radical consisting solely of carbon and hydrogen atoms, which can include fused, bridged, or spiral ring systems, having from three to twenty carbon atoms, preferably having from three to ten carbon atoms, and which is attached to the rest of the molecule by a single bond. Monocyclic cycloalkyl radicals include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic cycloalkyl radicals include, for example, adamantyl, norbornyl, dccalinyl,7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like. Unless otherwise stated specifically in the specification, a cycloalkyl group can be optionally substituted.

[0153] “Haloalkyl” refers to an alkyl radical, as defined above, that is substituted by one or more halo radicals, as defined above, e.g., trifluoromethyl, difluoromethyl, tri chloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl,1,2-dibromoethyl, and the like. Unless stated otherwise specifically in the specification, a haloalkyl group can be optionally substituted.

[0154] “Haloalkenyl” refers to an alkenyl radical, as defined above, that is substituted by one or more halo radicals, as defined above, e.g., 1 -fluoropropenyl, 1,1-difluorobutenyl, and the like. Unless stated otherwise specifically in the specification, a haloalkenyl group can be optionally substituted.

[0155] “Haloalkynyl” refers to an alkynyl radical, as defined above, that is substituted by one or more halo radicals, as defined above, e.g., 1-fluoropropynyl, 1 -fluorobutynyl, and the like. Unless stated otherwise specifically in the specification, a haloalkynyl group can be optionally substituted.

[0156] “Heterocyclyl,” “heterocyclic ring” or “heterocycle” refers to a stable 3- to 20-membered non-aromatic, partially aromatic, or aromatic ring radical which consists of two to twelve carbon atoms and from one to six heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur. Heterocyclycl or heterocyclic rings include heteroaryls as defined below. Unless stated otherwise specifically in the specification, theheterocyclyl radical can be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which can include fused, bridged, and spiral ring systems; and the nitrogen, carbon or sulfur atoms in the heterocyclyl radical can be optionally oxidized; the nitrogen atom can be optionally quaternized; and the heterocyclyl radical can be partially or fully saturated. Examples of such heterocyclyl radicals include, but are not limited to, aziridinyl, oextanyl, dioxolanyl, thienyl[l,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1 -oxo-thiomorpholinyl,1,1-dioxo-thiomorpholinyl, pyridine-one, and the like. The point of attachment of the heterocyclyl, heterocyclic ring, or heterocycle to the rest of the molecule by a single bond is through a ring member atom, which can be carbon or nitrogen. Unless stated otherwise specifically in the specification, a heterocyclyl group can be optionally substituted.

[0157] “Heterocyclylalkyl” refers to a radical of the formula -Rb-Rewhere Rb is an alkylene group as defined above and Reis a heterocyclyl radical as defined above. Unless stated otherwise specifically in the specification, a heterocyclylalkyl group can be optionally substituted.

[0158] “Heterocyclylalkenyl” refers to a radical of the formula -Rb-Rewhere Rb is an alkenylene group as defined above and Reis a heterocyclyl radical as defined above. Unless stated otherwise specifically in the specification, a heterocyclylalkenyl group can be optionally substituted.

[0159] “Heterocyclylalkynyl” refers to a radical of the formula -Rb-Rewhere Rb is an alkynylene group as defined above and Reis a heterocyclyl radical as defined above. Unless stated otherwise specifically in the specification, a heterocyclylalkynyl group can be optionally substituted.

[0160] ‘W-heterocyclyl” refers to a heterocyclyl radical as defined above containing at least one nitrogen and where the point of attachment of the heterocyclyl radical to the rest of the molecule is through a nitrogen atom in the heterocyclyl radical. Unless stated otherwise specifically in the specification, a < V-hclcrocyclyl group can be optionally substituted.

[0161] “Heteroaryl” refers to a 5- to 20-membered ring system radical one to thirteen carbon atoms and one to six heteroatoms selected from the group consisting of nitrogen,oxygen and sulfur, as the ring member. For purposes of this invention, the heteroaryl radical can be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which can include fused or bridged ring systems, wherein at least one ring containing a heteroatom ring member is aromatic. The nitrogen, carbon or sulfur atoms in the heteroaryl radical can be optionally oxidized and the nitrogen atom can be optionally quaternized. Examples include, but are not limited to, azepinyl, acridinyl, benzimidazolyl, benzothiazolyl, benzindolyl, benzodioxolyl, benzofuranyl, benzooxazolyl, benzothiazolyl, benzothiadiazolyl, benzo[Z?][l,4]dioxepinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl), benzotriazolyl, benzo[4,6]imidazo[l,2-a]pyridinyl, carbazolyl, cinnolinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, 1 -oxidopyridinyl,1 -oxidopyrimidinyl, 1-oxidopyrazinyl, 1 -oxidopyridazinyl, 1 -phenyl- 1 / / -pyrrolyl. phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrazolopyridine, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, and thiophenyl (z.e., thienyl). Unless stated otherwise specifically in the specification, a heteroaryl group can be optionally substituted.

[0162] “7V-heteroaryl” refers to a heteroaryl radical as defined above containing at least one nitrogen and where the point of attachment of the heteroaryl radical to the rest of the molecule is through a nitrogen atom in the heteroaryl radical. Unless stated otherwise specifically in the specification, an 7V-heteroaryl group can be optionally substituted.

[0163] “Thioalkyl” refers to a radical of the formula -SRawhere Rais an alkyl, alkenyl, or alkynyl radical as defined above containing one to twelve carbon atoms. Unless stated otherwise specifically in the specification, a thioalkyl group can be optionally substituted.

[0164] The term “substituted” used herein means any of the above groups (e.g., alkyl, alkylene, alkenyl, alkenylene, alkynyl, alkynylene, alkoxy, alkylamino, alkylcarbonyl, thioalkyl, aryl, aralkyl, carbocyclyl, cycloalkyl, cycloalkenyl, cycloalkynyl, cycloalkylalkyl, haloalkyl, heterocyclyl, A^-helerocyclyl, heterocyclylalkyl, heteroaryl, etc.) wherein at least one hydrogen atom is replaced by a bond to a non-hydrogen atoms such as, but not limited to: a halogen atom such as F, Cl, Br, and I; an oxygen atom in groups such as hydroxylgroups, alkoxy groups, and ester groups; a sulfur atom in groups such as thiol groups, thioalkyl groups, sulfone groups, sulfonyl groups, and sulfoxide groups; a nitrogen atom in groups such as amines, amides, alkylamines, dialkylamines, arylamines, alkylarylamines, diarylamines, N-oxides, imides, and enamines; a silicon atom in groups such as trialkylsilyl groups, dialkylarylsilyl groups, alkyldiarylsilyl groups, and triarylsilyl groups; and other heteroatoms in various other groups. “Substituted” also means any of the above groups in which one or more hydrogen atoms are replaced by a higher-order bond (e.g., a double- or triple -bond) to a heteroatom, such as oxygen in oxo, carbonyl, carboxyl, and ester groups; and nitrogen in groups such as imines, oximes, hydrazones, and nitriles. For example, “substituted” includes any of the above groups in which one or more hydrogen atoms are replacedwith -NRgRh, -NRgC(=O)Rh, -NRgC(=O)NRgRh, -NRgC(=O)ORh, -NRgSO2Rh, -OC( =O)NRgRh, -ORg, -SRg, -SORg, -SO2Rg, -OSO2Rg, -SO2ORg, =NSO2Rg, and -SO2NRgRh. “Substituted” also means any of the above groups in which one or more hydrogen atoms are replaced with -C(=O)Rg, -C(=O)ORg, -C(=O)NRgRh, -CH2SO2Rg, -CH2SO2NRgRh. In the foregoing, Rgand Rh are the same or different and independently hydrogen, alkyl, alkenyl, alkynyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkenyl, cycloalkynyl, cycloalkylalkyl, haloalkyl, haloalkenyl, haloalkynyl, heterocyclyl, A^-helcrocyclyl, heterocyclylalkyl, heteroaryl, A'-hcicroaryl and / or heteroarylalkyl. “Substituted” further means any of the above groups in which one or more hydrogen atoms are replaced by a bond to an amino, cyano, hydroxyl, imino, nitro, oxo, thioxo, halo, alkyl, alkenyl, alkynyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkenyl, cycloalkynyl, cycloalkylalkyl, haloalkyl, haloalkenyl, haloalkynyl, heterocyclyl, A-heterocyclyl, heterocyclylalkyl, heteroaryl, / -heteroaryl and / or heteroarylalkyl group. In addition, each of the foregoing substituents can also be optionally substituted with one or more of the above substituents.

[0165] As used herein, the symbol “? ” (hereinafter can be referred to as “a point of attachment bond”) denotes a bond that is a point of attachment between two chemical entities, one of which is depicted as being attached to the point of attachment bond and the other of which is not depicted as being attached to the point of attachment bond. Forexample,AH ” indicates that the chemical entity “A” is bonded to another chemicalentity via the point of attachment bond. Furthermore, the specific point of attachment to the non-depicted chemical entity can be specified by inference. For example, the compoundXA-|, wherein X is“5” infers that the point of attachment bond is the bond by which X is depicted as being attached to the phenyl ring at the ortho position relative to fluorine.

[0166] The term “molecular glue” refers to the small molecules of the present disclosure that possess the ability to promote interactions between proteins or other biomolecules that do not typically interact under normal cellular conditions. These molecules act as bridges or facilitators, bringing together proteins that may not naturally bind to each other, thereby influencing various cellular processes. Molecular glues have potential applications in modulating protein-protein interactions (PPI) and developing novel therapeutic agents. They can be utilized to target specific proteins involved in diseases such as cancer, neurodegenerative disorders, and infectious diseases.

[0167] The phrases "parenteral administration" and "administered parenterally" are art-recognized terms, and include modes of administration other than enteral and topical administration, such as injections, and include, without limitation, intravenous, intramuscular, intrapleural, intravascular, intrapericardial, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intra-articular, subcapsular, subarachnoid, intraspinal and intrastcmal injection and infusion.

[0168] The term "treating" is art-recognized and includes inhibiting a disease, disorder or condition in a subject, e.g., impeding its progress; and relieving the disease, disorder or condition, e.g., causing regression of the disease, disorder and / or condition. Treating the disease or condition includes ameliorating at least one symptom of the particular disease or condition, even if the underlying pathophysiology is not affected.

[0169] The term "preventing" is art- recognized and includes stopping a disease, disorder or condition from occurring in a subject, which may be predisposed to the disease, disorder and / or condition but has not yet been diagnosed as having it. Preventing a condition related to a disease includes stopping the condition from occurring after the disease has been diagnosed but before the condition has been diagnosed.

[0170] A "patient," "subject," or "host" to be treated by the compounds or methods described herein may mean either a human or non-human animal, such as a mammal, a fish, a bird, a reptile, or an amphibian. Thus, the subject of the herein disclosed methods can be a human, non-human primate, horse, pig, rabbit, dog, sheep, goat, cow, cat, guinea pig or rodent. The term does not denote a particular age or sex. Thus, adult and newborn subjects, as well as fetuses, whether male or female, are intended to be covered. In one aspect, the subject is a mammal. A patient refers to a subject afflicted with a disease or disorder.

[0171] The terms "prophylactic” or “therapeutic" treatment is art-recognized and includes administration to the host of one or more of the subject compositions. If it is administered prior to clinical manifestation of the unwanted condition (e.g., disease or other unwanted state of the host animal) then the treatment is prophylactic, i.e., it protects the host against developing the unwanted condition, whereas if it is administered after manifestation of the unwanted condition, the treatment is therapeutic (i.e., it is intended to diminish, ameliorate, or stabilize the existing unwanted condition or side effects thereof).

[0172] The term “PROTAC” (proteolysis targeting chimera) refers to a heterobifunctional small molecule composed of two active domains and a linker, capable of removing specific unwanted proteins. Rather than acting as a conventional enzyme inhibitor, a PROTAC works by inducing selective proteolysis. PROTACs can consist of two covalently linked protein-binding molecules: one (in many instances) capable of engaging an, and another that binds to a target protein meant for degradation. Recruitment of the E3 ligase to the target protein results in ubiquitination and subsequent degradation of the target protein by the proteasome. The concept was initially described by Deshaies and coworkers in 2001 (Skamoto, K. M. et al„ Proc. Natl. Acad. Sci. USA 98 (2001) 8554-8559).

[0173] The term “target protein ligand” or TPL is used to describe a small molecule which binds to a target protein or other protein or polypeptide of interest and places / presents that protein or polypeptide in proximity to an ubiquitin ligase such that degradation of the protein or polypeptide by ubiquitin ligase may occur. Non-limiting examples of small molecule target protein binding moieties include compounds targeting Tau protein. Such small molecule target protein binding ligands also include pharmaceutically acceptable salts, enantiomers, solvates and polymorphs of these compositions, as well as other small molecules that may target a protein of interest. These binding moieties are linked to TRIM21 targeting groups through linker groups.

[0174] The term “target protein” is used to describe a protein or polypeptide, which is a target for binding to a compound or target protein ligand as described herein and degradation by ubiquitin ligase hereunder.

[0175] The terms "therapeutic agent", "drug", "medicament" and "bioactive substance" are art-recognized and include molecules and other agents that are biologically, physiologically, or pharmacologically active substances that act locally or systemically in a patient or subject to treat a disease or condition. The terms include without limitation pharmaceutically acceptable salts thereof and prodrugs. Such agents may be acidic, basic, or salts; they may be neutral molecules, polar molecules, or molecular complexes capable of hydrogen bonding; they may be prodrugs in the form of ethers, esters, amides and the like that are biologically activated when administered into a patient or subject.

[0176] The phrase "therapeutically effective amount" or “pharmaceutically effective amount” is an art-recognized term. In certain embodiments, the term refers to an amount of a therapeutic agent that produces some desired effect at a reasonable benefit / risk ratio applicable to any medical treatment. In certain embodiments, the term refers to that amount necessary or sufficient to eliminate, reduce or maintain a target of a particular therapeutic regimen. The effective amount may vary depending on such factors as the disease or condition being treated, the particular targeted constructs being administered, the size of the subject or the severity of the disease or condition. One of ordinary skill in the art may empirically determine the effective amount of a particular compound without necessitating undue experimentation. In certain embodiments, a therapeutically effective amount of a therapeutic agent for in vivo use will likely depend on a number of factors, including: the rate of release of an agent from a polymer matrix, which will depend in part on the chemical and physical characteristics of the polymer; the identity of the agent; the mode and method of administration; and any other materials incorporated in the polymer matrix in addition to the agent.

[0177] The term " ED50" is art-recognized. In certain embodiments, ED50 means the dose of a drug, which produces 50% of its maximum response or effect, or alternatively, the dose, which produces a pre-determined response in 50% of test subjects or preparations. The term " LD50" is art-recognized. In certain embodiments, LD50 means the dose of a drug, which is lethal in 50% of test subjects. The term "therapeutic index" is an art-recognized term, which refers to the therapeutic index of a drug, defined as LD50 / ED50.

[0178] The terms " IC50," or “half maximal inhibitory concentration” is intended to refer to the concentration of a substance (e.g., a compound or a drug) that is required for 50% inhibition of a biological process, or component of a process, including a protein, subunit, organelle, ribonucleoprotein, etc.

[0179] " Optional" or "optionally" means that the subsequently described circumstance may or may not occur, so that the description includes instances where the circumstance occurs and instances where it does not. For example, the phrase "optionally substituted" means that a non-hydrogen substituent may or may not be present on a given atom, and, thus, the description includes structures wherein a non-hydrogen substituent is present and structures wherein a non-hydrogen substituent is not present.

[0180] Throughout the description, where compositions are described as having, including, or comprising, specific components, it is contemplated that compositions also consist essentially of, or consist of, the recited components. Similarly, where methods or processes are described as having, including, or comprising specific process steps, the processes also consist essentially of, or consist of, the recited processing steps. Further, it should be understood that the order of steps or order for performing certain actions is immaterial so long as the compositions and methods described herein remains operable. Moreover, two or more steps or actions can be conducted simultaneously.

[0181] All percentages and ratios used herein, unless otherwise indicated, are by weight.

[0182] Embodiments described herein relate to compounds and methods for targeted protein degradation (TPD), and particularly to E3 ubiquitin ligase tripartite motif 21 (TRIM21) targeted probes, molecular glue degraders, and bifunctional or proteolysis targeting chimeric compounds (PROTACs) for TPD. TPD utilizes molecular glues or PROTACs to eliminate disease-causing proteins by promoting their interaction with E3 ubiquitin ligases.

[0183] We identified PRLX-93936 and BMS-214662 as molecular glues that directly target the E3 ligase, TRIM21, to induce degradation of nucleoporin proteins, leading to inhibition of nuclear export and ultimately cell death. Loss of nucleoporins and nuclear export accounts for past observations in which BMS-214662 led to disrupted subcellular protein localization. Furthermore, the cytotoxicity of these compounds correlates withTRIM21 expression across hundreds of cancer cell lines, suggesting clinical re-evaluation of these agents in patients with TRIM21-high cancers.

[0184] Relative to recently-reported TRIM21 -targeting glues, these compounds and analogs based on these compounds display high cellular potency, creating new opportunities for targeted protein degradation via the design of additional glues and the design of PROTACs. Functionalization of PREX-93936 into PROTACs enabled selective degradation of multimeric proteins, such as those within biomolecular condensates, while sparing monomeric proteins. Such PROTACs can find utility as modulators of targeted ubiquitination of a variety of polypeptides and other proteins, which are then degraded and / or otherwise inhibited by the PROTACs described herein. An advantage of the compounds described herein is that a broad range of pharmacological activities is possible, consistent with the dcgradation / inhibition of targeted polypeptides from virtually any protein class or family. As aberrant protein assemblies cause diseases, such as autoimmunity, neurodegeneration, and cancer, our findings highlight the potential of TRIM21 -based multimer-selective degraders as a strategy to treat the direct causes of these diseases.

[0185] Advantageously, TRIM21 is the ideal E3 ligase for the development of protein aggregate PROTACs, especially in the context of tau aggregates. “Substrate clustering activation” of TRIM21 is what potentiates its unique competency in the development of aggregate-specific TPD relative to other E3 ligases. TRIM21’s E3 ligase activity is first activated and then enhanced when several TRIM21 homodimers associate via their RING domains at a protein target. Unlike E3 ligases, such as CRBN and VHL, non-intermolecular associated TRIM21 does not have E3 ligase activity until further TRIM21 dimers are recruited to a target. This can be leveraged for aggregate selective targeting of TPD pathological tau aggregates.

[0186] In some embodiments, a TRIM21 binding moiety (or ligand for TRIM21) or compound that is used for TPD can have the structure of formula (I):R4or a pharmaceutically acceptable salt, tautomer, or solvate thereof; whereinR1is absent, halogen, alkyl, haloalkyl, or alkoxy;R2aand R2bare each independently -H, halogen, alkyl, or haloalkyl, or alternatively R2aand R2btogether form a cycloalkyl or heterocyclyl;R3and R5are each independently -H, halogen, alkyl, haloalkyl, or oxo;R4is absent, -H, halogen, alkyl, haloalkyl, or -C(O)-alkyl optionally substituted with one or more halogen;R6and R7are each independently -H, halogen, alkyl, or haloalkyl;R8is absent, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, -alkylene-OH optionally substituted with one or more halogen, -CN, -C(O)-N(R10)2, -C(O)O-alkyl optionally substituted with one or more halogen, or carboxyl;R9is -H, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, or -O-alkylene-alkynyl optionally substituted with one or more halogen;each R10is independently -H, alkyl, or haloalkyl;X is N or O; oralternatively, one of R1, R2a, R2b, R3, R4, R5, R6, R8, or R9is substituted with a linker that is optionally linked to a target protein ligand; andwherein at least one of R1, R2a, R2b, R3, R4, R5, R6, R7, or R8is not absent or -H if X is N and R9is alkoxy, such as ethoxy.

[0187] In some embodiments, R1is absent, halogen, Ci-Cs alkyl, C1-C6 haloalkyl, or Ci-C6 alkoxy.

[0188] In some embodiments, R1is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, or ethoxy.

[0189] In some embodiments, R2aand R2bare each independently -H, halogen Ci-Ce alkyl, or Ci-Ce haloalkyl, or R2aand R2btogether form a C3-C6 cycloalkyl.

[0190] In other embodiments, one of R2aor R2bis -H, -F, -Cl, -Br, -I, methyl, ethyl, or isopropyl, and the other of R2aor R2bis -H, or R2aand R2btogether form cyclopropyl.

[0191] In some embodiments, R3is -H, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, or oxo.

[0192] In other embodiments, R3is -H, -F, -CF3, methyl, ethyl, isopropyl, or oxo.

[0193] In some embodiments, X is N and R4is -H, Ci-Ce alkyl, or -C(O)-(Ci-Ce alkyl).

[0194] In other embodiments, X is N and R4is -H, methyl, ethyl, isopropyl, -C(O)methyl, -C(O)ethyl, or -C(O)isopropyl.

[0195] In still other embodiments, X is O and R4is absent.

[0196] In some embodiments, R3is -H, halogen, Ci-Ce alkyl, or Ci-Ce haloalkyl.

[0197] In other embodiments, R5is -H, -F, -Cl, -Br, -I, -CF3, methyl, ethyl, or isopropyl.

[0198] In some embodiments, R6is -H, halogen, Ci-Ce alkyl, or Ci-Ce haloalkyl.

[0199] In other embodiments, R6is -H, -F, -Cl, -Br, -I, -CF3, methyl, ethyl, or isopropyl.

[0200] In some embodiments, R7is -H, halogen, Ci-Ce alkyl, or Ci-Ce haloalkyl.

[0201] In some embodiments, R7is -H, -F, -Cl, -Br, -I, -CF3, methyl, ethyl, or isopropyl.

[0202] In some embodiments, R8is absent, halogen, Ci-Ce alkyl, Ci-C6 haloalkyl, Ci- Ce alkoxy, -(Ci-Cealkylene)-OH, -CN, -C(O)-N(H)(Ci-C6alkyl), -C(O)O-(Ci-C6alkyl), or carboxyl.

[0203] In other embodiments, R8is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, ethoxy, propoxy, -methylene-OH, ethylene-OH, -propylene- OH, -CN, -C(O)-N(H)(methyl), -C(O)-N(H)(ethyl), -C(O)-N(H)(propyl), -C(O)O-(methyl), -C(O)O-(ethyl), -C(O)O-(propyl), or carboxyl.

[0204] In some embodiments, R9is -H, halogen, Ci-Ce alkyl, Ci-Ce alkoxy, or -O-(Ci-Ce alkylene)-alkynyl.

[0205] In other embodiments, R9is -H, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, ethoxy, propoxy, or -O-(methylene)-ethynyl.

[0206] In some embodiments, a compound of formula (I) does not have the structure of:or a pharmaceutically acceptable salt, tautomer, or solvate thereof.

[0207] In other embodiments, R1is absent, halogen, alkyl, haloalkyl, or alkoxy; R2ais alkyl or haloalkyl; R2bis -H; R3, R5, R6, and R7are each independently -H, halogen, alkyl, or haloalkyl; R4is -H; R8is absent, halogen, alkyl, haloalkyl, -alkylene-OH, -CN, -C(O)-N(R10)2, or -C(O)O-alkyl; R9is alkoxy or -O-alkylene-alkynyl; each R10is independently -H or alkyl; and X is N; or alternatively, one of R1, R2a, R2b, R3, R4, R5, R6, R8, or R9is substituted with a linker that is optionally linked to a target protein ligand.

[0208] In other embodiments, R1is absent, halogen, Ci-C8 alkyl, Ci-Ce haloalkyl, Ci-Ce alkoxy; R2ais Ci-Ce alkyl or Ci-Ce haloalkyl; R2bis -H; R3, R5, R6, and R7are each independently -H, halogen, Ci-Ce alkyl, or Ci-Ce haloalkyl; R4is -H; R8is absent, halogen, Ci-C6alkyl, Ci-C6haloalkyl, Ci-C6alkoxy, -(Ci-C6alkylene)-OH, -CN, -C(O)-N(H)(Ci-C6alkyl), -C(O)O-(Ci-Ce alkyl), or carboxyl; R9is Ci-Ce alkoxy or -O-(Ci-Ce alkylene)-alkynyl; and X is N; or alternatively, one of R1, R2a, R2b, R3, R4, R7, R6, R8, or R9is substituted with a linker that is optionally linked to a target protein ligand.

[0209] In other embodiments, R1is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, or ethoxy; R2ais methyl, ethyl, isopropyl, or -CF3; R2bis -H; R3, R5, R6, and R7are each independently -H, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, or -CF3; R4is -H; R8is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, ethoxy, propoxy, -methylene-OH, ethylene-OH, -propylene-OH, -CN, -C(O)-N(H)(methyl), -C(O)-N(H)(ethyl), -C(O)-N(H)(propyl), -C(O)O-(methyl), -C(O)O-(ethyl), -C(O)O-(propyl), or carboxyl; R9is methoxy, ethoxy, propoxy, or -O-(methylene)-ethynyl; and X is N; or alternatively, one of R1, R2a, R2b, R3, R4, R5, R6, R8, or R9is substituted with a linker that is optionally linked to a target protein ligand.

[0210] In some embodiments, the compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof includes an atropisomer of the compound offormula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof, wherein the atropisomer is substantially free of its opposite enantiomer or pharmaceutically acceptable salt, tautomer, or solvate thereof.

[0211] An atropisomer "substantially free" of its corresponding enantiomer means that a composition contains at least 90% by weight of one atropisomer, and 10% by weight or less of its stereoisomeric atropisomer. In some embodiments, the composition contains at least 95% by weight of one atropisomer and 5% by weight or less of its stereoisomer. In some embodiments, the composition contains at least 98% by weight of one atropisomer and 2% by weight or less of its stereoisomer. Alternatively, the relative amounts of the predominant isomer and any of the minor enantiomer is at least 9:1, or at least 19:1, or at least 98:2. In some embodiments, the composition contains at least 99% by weight of one atropisomer and 1% by weight or less of its stereoisomer. In some embodiments, the composition contains at least 99.5% by weight of one atropisomer and 0.5% by weight or less of its stereoisomer.

[0212] In some embodiments, the compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof is preferably a non-racemic mixture wherein the P isomer is the major component of the mixture. Typically such mixture will contain no more than about 10% of the M isomer, meaning the ratio of P to M isomers is at least about 9:1, and preferably less than 5% of the M isomer, meaning the ratio of P to M enantiomers is at least about 19:1. In some embodiments the compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof has less than 2% M enantiomer, meaning it has an enantiomeric excess of at least about 96%. In some embodiments, the compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof has an enantiomeric excess of at least 98%. In some embodiments, the compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof has an enantiomeric excess of at least 99%.

[0213] In other embodiments, the compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof is preferably a non-racemic mixture wherein the M isomer is the major component of the mixture. Typically such mixture will contain no more than about 10% of the P isomer, meaning the ratio of M to P isomers is at least about 9:1, and preferably less than 5% of the P isomer, meaning the ratio of M to P enantiomers is at least about 19:1. In some embodiments the compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof has less than 2% P enantiomer, meaning it has anenantiomeric excess of at least about 96%. In some embodiments, the compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof has an enantiomeric excess of at least 98%. In some embodiments, the compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof has an enantiomeric excess of at least 99%.

[0214] An atropisomer which is present "in excess" of its corresponding enantiomer or an "enantioenriched mixture" means that the atropisomer is present in an amount greater than its enantiomer, making the atropisomer mixture optically active. Typically this means the compound present "in excess" predominates by at least a 60 / 40 ratio over its enantiomer.

[0215] In other embodiments, a TRIM21 binding moiety (or ligand for TRIM21) or compound that is used for TPD can have the structure of formula (IA):R4(IA) or a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinR1is absent, halogen, alkyl, haloalkyl, or alkoxy;R2ais -H, halogen, alkyl, or haloalkyl;R3is -H, halogen, alkyl, or haloalkyl;R4is -H, halogen, alkyl, haloalkyl, or -C(O)-alkyl optionally substituted with one or more halogen;R8is absent, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, -alkylene-OH optionally substituted with one or more halogen, -CN, -C(O)-N(R10)2, -C(O)O-alkyl optionally substituted with one or more halogen, or carboxyl;each R10is independently -H, alkyl, or haloalkyl; oralternatively, one of R1, R2a, R3, R4, or R8is substituted with a linker that is optionally linked to a target protein ligand; andwherein at least one of R1, R2a, R3, R4, or R8is not absent or -H.

[0216] In some embodiments, R1is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, or Ci-Ce alkoxy.

[0217] In other embodiments, R1is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, or ethoxy.

[0218] In some embodiments, R2ais -H, halogen, C1-C.6 alkyl, or C1-C.6 haloalkyl.

[0219] In other embodiments, R2ais -H, -F, -CF3, methyl, ethyl, or isopropyl.

[0220] In some embodiments, R3is -H, halogen, Ci-C6 alkyl, or Ci-C6 haloalkyl.

[0221] In other embodiments, R is -H, -F, -CF3, methyl, ethyl, or isopropyl.

[0222] In some embodiments, R4is -H, Ci-Ce alkyl, or -C(O)-(Ci-C6 alkyl).

[0223] In other embodiments, R4is -H, methyl, ethyl, isopropyl, -C(O)methyl, -C(O)ethyl, or -C(O)isopropyl.

[0224] In some embodiments, R8is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, Ci-Ce alkoxy, -(Ci-Cealkylene)-OH, -CN, -C(O)-N(H)(Ci-Ce alkyl), -C(O)O-(Ci-Ce alkyl), or carboxyl.

[0225] In other embodiments, R8is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, ethoxy, propoxy, -methylene-OH, ethylene-OH, -propylene-OH, -CN, -C(O)-N(H)(methyl), -C(O)-N(H)(ethyl), -C(O)-N(H)(propyl), -C(O)O-(methyl),-C(O)O-(ethyl), -C(O)O-(propyl), or carboxyl.

[0226] In some embodiments, the compound of formula (I A) or pharmaceutically acceptable salt, tautomer, or solvate thereof can include an atropisomer of the compound of formula (IA) or pharmaceutically acceptable salt, tautomer, or solvate thereof, wherein the atropisomer is substantially free of its opposite enantiomer or pharmaceutically acceptable salt, tautomer, or solvate thereof.

[0227] In some embodiments, the atropisomer is a P isomer.

[0228] In other embodiments, a TRIM21 binding moiety (or ligand for TRIM21) or compound that is used for TPD can have the structure of formula (IB):R4(IB) or a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinR1is absent, halogen, alkyl, haloalkyl, or alkoxy;R2ais alkyl or haloalkyl;R3is -H, halogen, alkyl, or haloalkyl;R4is -H, halogen, alkyl, haloalkyl, or -C(O)-alkyl optionally substituted with one or more halogen;R8is absent, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, -alkylene-OH optionally substituted with one or more halogen, -CN, -C(O)-N(R10)2, -C(O)O-alkyl optionally substituted with one or more halogen, or carboxyl; and each R10is independently -H, alkyl, or haloalkyl; oralternatively, one of R1, R2a, R3, R4, or R8is substituted with a linker that is optionally linked to a target protein ligand.

[0229] In some embodiments, R1is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, or Ci-C6 alkoxy.

[0230] In other embodiments, R1is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, or ethoxy.

[0231] In some embodiments, R2ais Ci-Ce alkyl or Ci-Ce haloalkyl.

[0232] In other embodiments, R2ais -CF3, methyl, ethyl, or isopropyl.

[0233] In some embodiments, R3is -H, halogen, Ci-Ce alkyl, Ci-Ce or haloalkyl.

[0234] In other embodiments, R3is -H, -F, -CF3, methyl, ethyl, or isopropyl.

[0235] In some embodiments, R4is -H, Ci-Ce alkyl, or -C(O)-(Ci-Ce alkyl).

[0236] In other embodiments, R4is -H, methyl, ethyl, isopropyl, -C(O)methyl, -C(O)ethyl, or -C(O)isopropyl.

[0237] In some embodiments, R8is absent, halogen, Ci-C8 alkyl, Ci-Ce haloalkyl, Ci- Ce alkoxy, -(Ci-Cealkylene)-OH, -CN, -C(O)-N(H)(Ci-Ce alkyl), -C(O)O-(Ci-Ce alkyl), or carboxyl.

[0238] In other embodiments, R8is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, ethoxy, propoxy, -methylene-OH, ethylene-OH, -propylene- OH, -CN, -C(O)- N(H)(methyl), -C(O)-N(H)(ethyl), -C(O)-N(H)(propyl), -C(O)O-(methyl), -C(O)O-(ethyl), -C(O)O-(propyl), or carboxyl.

[0239] In some embodiments, the compound of formula (I A) or pharmaceutically acceptable salt, tautomer, or solvate thereof can include an atropisomer of the compound of formula (IA) or pharmaceutically acceptable salt, tautomer, or solvate thereof, wherein the atropisomer is substantially free of its opposite enantiomer or pharmaceutically acceptable salt, tautomer, or solvate thereof.

[0240] In some embodiments, the atropisomer is a P isomer.

[0241] In other embodiments, a TRIM21 binding moiety (or ligand for TRIM21) or compound that is used for TPD can include an atropisomer of formula (IC):(IC) or a pharmaceutically acceptable salt, tautomer, or solvate thereof:whereinR1is absent, halogen, alkyl, haloalkyl, or alkoxy;R2ais alkyl or haloalkyl;R8is absent, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, -alkylene-OH optionally substituted with one or more halogen, -CN, -C(O)-N(R10)2, -C(O)O-alkyl optionally substituted with one or more halogen, or carboxyl;each R10is independently -H, alkyl, or haloalkyl; oralternatively, one of R1, R2a, or R8is substituted with a linker that is optionally linked to a target protein ligand; andwherein the compound of formula (IC) or pharmaceutically acceptable salt, tautomer, or solvate thereof is substantially free of its opposite enantiomer or pharmaceutically acceptable salt, tautomer, or solvate thereof.

[0242] In some embodiments, R1is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, or Ci-C6alkoxy.

[0243] In other embodiments, R1is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, or ethoxy.

[0244] In some embodiments, R2ais Ci-Ce alkyl or Ci-Ce haloalkyl.

[0245] In other embodiments, R2ais -CF3, methyl, ethyl, or isopropyl.

[0246] In some embodiments, R8is absent, halogen, C1-C6 alkyl, Ci-Ce haloalkyl, Ci-Ce alkoxy, -(Ci-C6alkylene)-OH, -CN, -C(O)-N(H)(Ci-C6alkyl), -C(O)O-(Ci-C6alkyl), or carboxyl.

[0247] In other embodiments, R8is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, ethoxy, propoxy, -methylene-OH, ethylene-OH, -propylene- OH, -CN, -C(O)- N(H)(methyl), -C(O)-N(H)(ethyl), -C(O)-N(H)(propyl), -C(O)O-(methyl), -C(O)O-(ethyl), -C(O)O-(propyl), or carboxyl.

[0248] In some embodiments, the atropisomer is a P isomer.

[0249] In some embodiments, the compound of formula (IC) or pharmaceutically acceptable salt, tautomer, or solvate thereof has a shorter retention time or elutes faster by chiral separation than its opposite enantiomer or pharmaceutically acceptable salt, tautomer, or solvate thereof.

[0250] In other embodiments, an atropisomer of formula (IC) can include a compound of formula (IC1):H NR1R8CR8b(IC1) or a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinR1is absent, halogen, alkyl, haloalkyl, or alkoxy;R2ais alkyl or haloalkyl;one of R8a, R8b, and R8cis -H, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, -alkylene-OH optionally substituted with one or more halogen, -CN, -C(O)-N(R10)2, -C(O)O-alkyl optionally substituted with one or more halogen, or carboxyl and the other two of R8a, R8b, and R8care -H;each R10is independently -H, alkyl, or haloalkyl; oralternatively, one of R1, R2a, R8a, R8b, or R8cis substituted with a linker that is optionally linked to a target protein ligand; andwherein the compound of formula (IC1) or pharmaceutically acceptable salt, tautomer, or solvate thereof is substantially free of its opposite enantiomer or pharmaceutically acceptable salt, tautomer, or solvate thereof.

[0251] In some embodiments, R1is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, or Ci-C6alkoxy.

[0252] In other embodiments, R1is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, or ethoxy.

[0253] In some embodiments, R2ais Ci-Ce alkyl or Ci-Ce haloalkyl.

[0254] In other embodiments, R2ais -CF3, methyl, ethyl, or isopropyl.

[0255] In some embodiments, one of R8a, R8b, and R8cis -H, halogen, Ci-Ce alkyl, Ci- Ce haloalkyl, Ci-Ce alkoxy, -(Ci-Ce alkylene)-OH, -CN, -C(O)-N(H)(Ci-Ce alkyl), -C(O)O- (Ci-Ce alkyl), or carboxyl, and the other two of R8a, R8b, and R8cis -II.

[0256] In other embodiments, one of R8a, RSb, and R8cis -H, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, ethoxy, propoxy, -methylene-OH, ethylene-OH, -propylene -OH, -CN, -C(O)-N(H)(methyl), -C(O)-N(H)(ethyl), -C(O)-N(H)(propyl), -C(O)O-(methyl), -C(O)O-(ethyl), -C(O)O-(propyl), or carboxyl, and the other two of R8a, R8a, and R8ais -H.

[0257] In some embodiments, the atropisomer is a P isomer, and the atropisomer is substantially free of its opposite enantiomer or pharmaceutically acceptable salt, tautomer, or solvate thereof.

[0258] In some embodiments, the compound of formula (IC1) or pharmaceutically acceptable salt, tautomer, or solvate thereof has a shorter retention time or elutes faster by chiral separation than its opposite enantiomer or pharmaceutically acceptable salt, tautomer, or solvate thereof.

[0259] In some embodiments, an atropisomer of formula (IC1) can have the structure of:or a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinR1is absent, halogen, alkyl, haloalkyl, or alkoxy;R2ais alkyl or haloalkyl;R8ais -H, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, -alkylene-OH optionally substituted with one or more halogen, -CN, -C(O)-N(R10)2, -C(O)O-alkyl optionally substituted with one or more halogen, or carboxyl;each R10is independently -H, alkyl, or haloalkyl; oralternatively, one of R1, R2a, or R8ais substituted with a linker that is optionally linked to a target protein ligand.

[0260] In other embodiments, an atropisomer of formula (IC1) can have the structure of:HR8bor a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinR1is absent, halogen, alkyl, haloalkyl, or alkoxy;R2ais alkyl or haloalkyl;R8bis -H, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, -alkylene-OH optionally substituted with one or more halogen, -CN, -C(O)-N(R10)2, -C(O)O-alkyl optionally substituted with one or more halogen, or carboxyl;each R10is independently -H, alkyl, or haloalkyl; oralternatively, one of R1, R2a, or R8bis substituted with a linker that is optionally linked to a target protein ligand.

[0261] In some embodiments, an atropisomer of formula (IC 1 ) can have the structureor a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinR1is absent, halogen, alkyl, haloalkyl, or alkoxy;R2ais alkyl or haloalkyl;R8cis -H, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, -alkylene-OH optionally substituted with one or more halogen, -CN, -C(O)-N(R10)2, -C(O)O-alkyl optionally substituted with one or more halogen, or carboxyl;each R10is independently -H, alkyl, or haloalkyl; oralternatively, one of R1, R2a, or R8cis substituted with a linker that is optionally linked to a target protein ligand.

[0262] In other embodiments, an atropisomer of formula (IC) can include a compound of formula (IC2):HNR1aR2a NR1bR1Rid(IC2) or a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinone Rla, Rlb, Rlc, or Rldis -H, halogen, alkyl, haloalkyl, or alkoxy, and the other three of Rla, Rlb, Rlc, or Rldis -H;R2ais alkyl or haloalkyl;R8is absent, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, -alkylene-OH optionally substituted with one or more halogen, -CN, -C(O)-N(R10)2, -C(O)O-alkyl optionally substituted with one or more halogen, or carboxyl;each R10is independently -H, alkyl, or haloalkyl; oralternatively, one of Rla, Rlb, Rlc, Rld, R2a, or R8is substituted with a linker that is optionally linked to a target protein ligand; andwherein the compound of formula (IC2) or pharmaceutically acceptable salt, tautomer, or solvate thereof is substantially free of its opposite enantiomer or pharmaceutically acceptable salt, tautomer, or solvate thereof.

[0263] In some embodiments, one Rla, Rlb, Rlc, or Rldis -H, halogen, Ci-C.6 alkyl, Ci-Ce haloalkyl, or Ci-Ce alkoxy, and the other three of Rla, Rlb, Rlc, or Rldis -H.

[0264] In other embodiments, one Rla, Rlb, Rlc, or Rldis -H, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, or ethoxy, and the other three of Rla, Rlb, Rlc, or Rldis -H.

[0265] In some embodiments, R2ais C1-C6 alkyl or Ci-Ce haloalkyl.

[0266] In other embodiments, R2ais -CF3, methyl, ethyl, or isopropyl.

[0267] In some embodiments, R8is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, Ci-C6alkoxy, -(Ci-C6alkylene)-OH, -CN, -C(O)-N(H)(CI-C6alkyl), -C(O)O-(Ci-C6alkyl), or carboxyl.

[0268] In other embodiments, R8is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, ethoxy, propoxy, -methylene-OH, ethylene-OH, -propylene-OH, -CN, -C(O)-N(H)(methyl), -C(O)-N(H)(ethyl), -C(O)-N(H)(propyl), -C(O)O-(methyl), -C(O)O-(ethyl),-C(O)O-(propyl), or carboxyl.

[0269] In some embodiments, the atropisomer is a P isomer, and the atropisomer is substantially free of its opposite enantiomer or pharmaceutically acceptable salt, tautomer, or solvate thereof.

[0270] In some embodiments, the compound of formula (IC2) or pharmaceutically acceptable salt, tautomer, or solvate thereof has a shorter retention time or elutes faster by chiral separation than its opposite enantiomer or pharmaceutically acceptable salt, tautomer, or solvate thereof.

[0271] In some embodiments, an atropisomer of formula (IC2) can have the structure of:H NIs;R1aR2a NNT ®N, J- R8or a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinRlais -H, halogen, alkyl, haloalkyl, or alkoxy;R2ais alkyl or haloalkyl;R8is absent, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, -alkylene-OH optionally substituted with one or more halogen, -CN, -C(O)-N(R10)2, -C(O)O-alkyl optionally substituted with one or more halogen, or carboxyl;each R10is independently -H, alkyl, or haloalkyl; oralternatively, one of Rla, R2a, or R8is substituted with a linker that is optionally linked to a target protein ligand.

[0272] In some embodiments, an atropisomer of formula (IC2) can have the structure of:R1bor a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinRlbis -H, halogen, alkyl, haloalkyl, or alkoxy;R2ais alkyl or haloalkyl;R8is absent, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, -alkylene-OH optionally substituted with one or more halogen, -CN, -C(O)-N(R10)2, -C(O)O-alkyl optionally substituted with one or more halogen, or carboxyl;each R10is independently -H, alkyl, or haloalkyl; oralternatively, one of Rlb, R2a, or R8is substituted with a linker that is optionally linked to a target protein ligand.

[0273] In some embodiments, an atropisomer of formula (IC2) can have the structure of:T <?N..8or a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinRlcis -H, halogen, alkyl, haloalkyl, or alkoxy;R2ais alkyl or haloalkyl;R8is absent, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, -alkylene-OH optionally substituted with one or more halogen, -CN, -C(O)-N(R10)2, -C(O)O-alkyl optionally substituted with one or more halogen, or carboxyl;each R10is independently -H, alkyl, or haloalkyl; oralternatively, one of Rlc, R2a, or R8is substituted with a linker that is optionally linked to a target protein ligand.

[0274] In some embodiments, an atropisomer of formula (IC2) can have the structure of:>2a NT0N.,8or a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinRldis -H, halogen, alkyl, haloalkyl, or alkoxy;R2ais alkyl or haloalkyl;R8is absent, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, -alkylene-OH optionally substituted with one or more halogen, -CN, -C(O)-N(R10)2, -C(O)O-alkyl optionally substituted with one or more halogen, or carboxyl;each R10is independently -H, alkyl, or haloalkyl; oralternatively, one of Rld, R2a, or R8is substituted with a linker that is optionally linked to a target protein ligand.

[0275] Other embodiments relate to a compound of formula (I) selected from:0 o oor a pharmaceutically acceptable salt, tautomer, or solvate thereof.

[0276] Still other embodiments relate to a compound of formula (I) selected from:H H H HNNN NN N N NcXiOcCOcCC' Il? ’ * H H H HNN N NN N N NO O O O0 0 0pharmaceutically acceptable salt, tautomer, or solvate thereof.

[0277] In other embodiments, a TRIM21 binding moiety (or ligand for TRIM21) or compound that is used for TPD can have the structure of formula (II):(II) or a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinR11is -H, alkyl, or haloalkyl;R12and R13are each independently absent, -CN, alkyl, haloalkyl, haloalkenyl, alkynyl, or -alkylene-alkynyl optionally substituted with one or more halogen;R14is cycloalkyl, heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with R17;R15and R16are each independently absent, halogen, alkyl, or haloalkyl;R17is halogen, alkyl, or alkoxy;R18is -H, alkyl, or haloalkyl;X1is S(O)n, CH2, or CO;n is 0, 1, or 2;alternatively, one of R11, R12, R13, R14, R15, R16, R17, or R18is substituted with a linker that is optionally linked to a target protein ligand; andwherein at least one of R12and / or R13is not -CN if X1is S(O)2 and R14is a thiophenyl group.

[0278] In some embodiments, R11is H, Ci-Ce alkyl, or Ci-Cehaloalkyl.

[0279] In other embodiments, R11is -H, methyl, or ethyl, preferably -H.

[0280] In some embodiments, R12and R13are each independently absent, -CN, Ci-Ce alkyl, Ci-Cehaloalkyl, C2-C,6haloalkenyl, C2-C.6 alkynyl, -(C1-C5 alkylene)-(C,2-C6 alkynyl).

[0281] In other embodiments, R12and R13are each independently absent, -CN, or ethynyl.

[0282] In some embodiments, R14is Ca-Cs cycloalkyl, heterocyclyl having 5 to 10 ring atoms, C6-C10 aryl, or heteroaryl having 5 to 10 ring atoms, each of which is optionally substituted with R17.

[0283] In other embodiments, R14is thiophcnyl, phenyl, isoxazolyl, or thiazolyl, each of which is optionally substituted with R17.

[0284] In some embodiments, R15and R16are each independently absent, halogen, Ci-Ce alkyl, or Ci-Ce haloalkyl.

[0285] In some embodiments, R17is halogen, Ci-Ce alkyl, or Ci-Ce alkoxy.

[0286] In some embodiments, R18is -H.

[0287] In some embodiments, X1is S(O)2.

[0288] In some embodiments, one of R11, R12, R13, R14, R13, R16, R17, or R18is substituted with a linker that is linked to the target protein ligand.

[0289] Other embodiments relate to a compound of formula (II) selected from:CN CN NCN N HN NH

[0290] In some embodiments, a compound of formula (I), (IA), (IB), (IC), or (II), or a pharmaceutically acceptable salt, tautomer, or solvate thereof can be used as a molecular glue degrader that binds TRIM21 and a variety of polypeptides and other proteins, such as a nucleoporin, e.g., NUP214 and NUP88, to modulate targeted ubiquitination of the polypeptides and other proteins. The compound of formula (I), (IA), (IB), (IC), or (II) or a pharmaceutically acceptable salt, tautomer, or solvate thereof can direct TRIM21 to the nuclear pore to induce proteasomal degradation of multiple nucleoporin proteins, leading to inhibition of nuclear export and ultimately cell death.

[0291] In other embodiments, a compound of formula (I), (IA), (IB), (IC), or (II) or a pharmaceutically acceptable salt, tautomer, or solvate thereof can be linked, joined, or coupled directly or indirectly to a target protein ligand that binds a target protein to form a PROTAC. In certain embodiments, a compound of formula (I), (IA), (IB), (IC), or (II) or a pharmaceutically acceptable salt, tautomer, or solvate thereof, and the target protein ligand are joined or coupled via a chemical linker to form the PROTAC. The compound of formula (I), (IA), (IB), (IC), or (II), or a pharmaceutically acceptable salt, tautomer, or solvate thereof can recognize TRIM21, and the target protein ligand recognizes a target protein, and the interaction of the respective moieties with their targets facilitates degradation of the target protein by placing the target protein in proximity to TRIM21.

[0292] In certain embodiments, the PROTACs as described herein can include multiple target protein ligands (targeting the same or different protein targets), multiple compounds of formula (I), (IA), (IB), (IC), or (II) or a pharmaceutically acceptable salt, tautomer, or solvate thereof that bind specifically TRIM21. In any of the aspects of embodiments described herein, the target protein ligands and compounds of formula (I), (IA), (IB), (IC), or (II), or a pharmaceutically acceptable salt, tautomer, or solvate thereof can be coupled directly or via one or more chemical linkers or a combination thereof.

[0293] In some embodiments, a PROTAC that includes a compound of formula (I) or a pharmaceutically acceptable salt, tautomer, or solvate thereof linked to target protein ligand can have the structure of:R4(I) or a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinR1is absent, halogen, alkyl, haloalkyl, or alkoxy;R2aand R2bare each independently -H, halogen, alkyl, or haloalkyl, or alternatively R2aand R2btogether form a cycloalkyl or heterocyclyl;R and R5are each independently -H, halogen, alkyl, haloalkyl, or oxo;R4is absent -H, halogen, alkyl, haloalkyl, or -C(O)-alkyl optionally substituted with one or more halogen;R6and R7are each independently -H, halogen, alkyl, or haloalkyl;R'sis absent, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, -alkylene-OH optionally substituted with one or more halogen, -CN, -C(O)-N(R10)2, -C(O)O-alkyl optionally substituted with one or more halogen, or carboxyl;R9is -H, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, or -O-alkylene-alkynyl optionally substituted with one or more halogen;each R10is independently -H, alkyl, or haloalkyl;X is N or O; andwherein one of R1, R2a, R2b, R3, R4, R5, R6, R8, R9is substituted with a linker; anda target protein ligand is linked to the linker.

[0294] In some embodiments, R1is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, or Ci-Ce alkoxy.

[0295] In some embodiments, R2aand R2bare each independently -H, halogen Ci-Ce alkyl, or Ci-Ce haloalkyl, or R2aand R2btogether form a Cj-Ce cycloalkyl.

[0296] In some embodiments, R is -H, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, or oxo.

[0297] In some embodiments, X is N and R4is -H, Ci-Ce alkyl, or -C(O)-(Ci-Ce alkyl).

[0298] In some embodiments, R3is -H, halogen, Ci-Ce alkyl, or Ci-Ce haloalkyl.

[0299] In some embodiments, R6is -H, halogen, Ci-Ce alkyl, or Ci-Ce haloalkyl.

[0300] In some embodiments, R7is -H, halogen, Ci-Ce alkyl, or Ci-Ce haloalkyl.

[0301] In some embodiments, R8is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, Ci-Ce alkoxy, -(Ci-Cealkylene)-OH, -CN, -C(O)-N(H)(Ci-Ce alkyl), -C(O)O-(Ci-Ce alkyl), or carboxyl.

[0302] In some embodiments, R9is -H, Ci-Ce alkyl, Ci-Ce alkoxy, or -O-(Ci-Ce alkylene)-alkynyl.

[0303] In other embodiments, a PROTAC that includes a compound of formula (I) or a pharmaceutically acceptable salt, tautomer, or solvate thereof linked to a target protein ligand can include the formula selected from:or solvate thereof;whereinR1is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, or Ci-Ce alkoxy;R2ais Ci-C6alkyl or Ci-C6haloalkyl;R8is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, Ci-Ce alkoxy, -(Ci-Ce alkylene)-OH, -CN, -C(O)-N(H)(Ci-Ce alkyl), -C(O)O-(Ci-Ce alkyl), or carboxyl;R9is -H, Ci-Ce alkyl, Ci-Ce alkoxy, or -O-(Ci-Ce alkylene)-alkynyl;L is the linker; andTPL is the target protein ligand.

[0304] In still other embodiments, a PROTAC that includes a compound of formula (I) or a pharmaceutically acceptable salt, tautomer, or solvate thereof linked to a target protein ligand can include the formula selected from:or a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinR1is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, or Ci-Ce alkoxy;R2ais Ci-Ce alkyl or Ci-Ce haloalkyl;R8is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, Ci-Ce alkoxy, -(Ci-Ce alkylene)-OH, -CN, -C(O)-N(H)(CI-C6alkyl), -C(O)O-(Ci-C6alkyl), or carboxyl;L is the linker; and1'PL is the target protein ligand.

[0305] In some embodiments, the PROTAC including the compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof is an atropisomer orpharmaceutically acceptable salt, tautomer, or solvate thereof that is substantially free of its opposite enantiomer or pharmaceutically acceptable salt, tautomer, or solvate thereof.

[0306] In some embodiments, the atropisomer is a P isomer.

[0307] In some embodiments, the linker includes an aliphatic linker, a cyclic linker, or a polyethylene glycol linker.

[0308] In other embodiments, a PROTAC that includes a compound of formula (II) or a pharmaceutically acceptable salt, tautomer, or solvate thereof linked to target protein ligand can have the structure of:R15(II) or a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinR11is H, alkyl, or haloalkyl;R12and R13are each independently absent, -CN, alkyl, haloalkyl, haloalkenyl, alkynyl, or -alkylene-alkynyl optionally substituted with one or more halogen;R14is cycloalkyl, heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with R17;R15and R16are each independently absent, halogen, alkyl, or haloalkyl;R17is halogen, alkyl, or alkoxy;X1is S(O)n, CH2, or CO;n is 0, 1, or 2;wherein one R11, R12, R13, R14, R15, R16, or R17is substituted with a linker, and a target protein ligand is linked to the linker.

[0309] In some embodiments, R11is H, Ci-C.6 alkyl, Ci-Ce haloalkyl, preferably -H.

[0310] In some embodiments, R12and R13are each independently absent, -CN, Ci-Ce alkyl, Ci-Ce haloalkyl, C2-C6 haloalkenyl, C2-C6 alkynyl, -(Ci-Ce alkylene)-(C2-Ce alkynyl).

[0311] In some embodiments, R14is Cs-Cs cycloalkyl, heterocyclyl having 5 to 10 ring atoms, Ce-Cio aryl, or heteroaryl having 5 to 10 ring atoms, each of which is optionally substituted with R17preferably R14is thiophenyl, phenyl, isoxazolyl, or thiazolyl, each of which is optionally substituted with R17.

[0312] In some embodiments, X1is S(O)2.

[0313] In some embodiments, R13and R16are each independently absent, halogen, Ci- C6 alkyl, or Ci-Cc haloalkyl.

[0314] In some embodiments, R17is halogen, Ci-Ce alkyl, or Ci-Ce alkoxy.

[0315] In some embodiments, a PROTAC that includes a compound of formula (II) or a pharmaceutically acceptable salt, tautomer, or solvate thereof linked to a target protein ligand can include the formula selected from:tautomer, or solvate thereof;whereinR14is C.3-Cs cycloalkyl, heterocyclyl having 5 to 10 ring atoms, Ce-Cio aryl, or heteroaryl having 5 to 10 ring atoms, each of which is optionally substituted with R17;R15and R16are each independently absent, halogen, Ci-Ce alkyl, or Ci-Ce haloalkyl;R17is halogen, alkyl, or alkoxy;L1is the linker; andTPL is the target protein ligand.

[0316] In other embodiments, a PROTAC that includes a compound of formula (II) or a pharmaceutically acceptable salt, tautomer, or solvate thereof linked to a target protein ligand can include the formula selected from:CN CN CNpharmaceutically acceptable salt, tautomer, or solvate thereof;whereinL1is the linker; andTPL is the target protein ligand.

[0317] In some embodiments, the linker (e.g., L or L1) that chemically links the compounds of formula (I), (IA), (IB), (IC), or (II), or a pharmaceutically acceptable salt, tautomer, or solvate thereof to the target protein ligand can include one or more covalently connected structural units of A (e.g.,- i ■■■ Aq-), wherein Ai is a group coupled to at least one of a compounds of formula (I), (IA), (IB), (IC), or (II), or a pharmaceutically acceptable salt, tautomer, or solvate thereof, a target protein ligand, or a combination thereof. In certain embodiments, Ai links a compound of formula (I), (IA), (IB), (IC), or (II), or a pharmaceutically acceptable salt, tautomer, or solvate thereof, target protein ligand, or acombination thereof directly to another compound of formula (I), (I A), (IB), (IC), or (II), or a pharmaceutically acceptable salt, tautomer, or solvate thereof, target protein ligand or combination thereof. In other embodiments, Ai links a compound of formula (I), formula (II), or a pharmaceutically acceptable salt, tautomer, or solvate thereof, target protein ligand or a combination thereof indirectly to another compound of formula (I), formula (II), or a pharmaceutically acceptable salt, tautomer, or solvate thereof, target protein ligand or combination thereof through Aq.

[0318] In certain embodiments, Ai to Aqare, each independently, a bond, CRL1RL2, O, s, so, so2, NRL3, SO2NRL3, SONRL3, CONRL3, NRL3CONRL4, NRL3SO2NRL4, CO.CRL1=CRL2, C=C, SiRL1RL2, P(O)RL1, P(O)ORL1, NRL3C(=NCN)NRL4, NRL3C(=NCN), NRL3C(=CNO2)NRL4, C3-C11 cycloalkyl optionally substituted with 0-6 RL1and / or RL2groups, C3-C11 hctcrocyclyl optionally substituted with 0-6 RL1and / or RL2groups, aryl optionally substituted with 0-6 RL1and / or RL2groups, heteroaryl optionally substituted with 0-6 RL1and / or RL2groups, where RL1or RL2, each independently, can be linked to other A groups to form cycloalkyl and / or heterocyclyl moiety which can be further substituted with 0-4 RL5groups; wherein RL1, RL2, RL3, R1and RL5are, each independently, H, halo, Ci-Cs alkyl. O(Ci-C8alkyl), S(Ci-C8alkyl), NH(Ci-Cs alkyl), N(CI-C8alkyl)2, C3-C11 cycloalkyl, aryl, heteroaryl, C3-C11 heterocyclyl, O(C3-Cn cycloalkyl), S(Ci-C8cycloalkyl), NH(CI-C8cycloalkyl), N(CI-C8cycloalkyl)2, N(Ci-Cs cycloalkyl)( Ci-Cs alkyl), OH, NH2, SH, SO2(Ci-C8alkyl), P(O)(OCi-C8alkyl)( Ci-C8alkyl), P(O)(OCi-C8alkyl)2, CC— Ci-C8alkyl, CCH, CH=CH(C1-C8alkyl), C(Ci-C8alkyl)=CH(Ci-C8alkyl), C(Ci-C8alkyl)=C(Ci-C8alkyl)2, Si(OH)3, Si(Ci-C8alkyl)3, Si(OH)( Ci-C8alkyl)2, CO(Ci-C8alkyl), CO2H, halogen, CN, CF3, CHF2, CH2F, NO2, SF5, SO2NH(CI-C8alkyl), SO2N(CI-C8alkyl).sub.2, SONH(CI-C8alkyl), SON(CI-C8alkyl)2, CONH(CI-C8alkyl), CON(CI-C8alkyl)2, N(CI-C8alkyl)CONH(Ci-Cs alkyl), N(CI-C8alkyl)CON(Ci-C8alkyl)2, NHCONH(CI-C8alkyl), NHCON(CI-C8alkyl)2, NHCONH2, N(CI-C8alkyl)SO2NH(Ci-C8alkyl), N(CI-C8alkyl) SO2N(CI-C8)2, NH SO2NH(CI-C8alkyl), NH SO2N(CI-C8alkyl)2, NH SO2NH2.

[0319] In some embodiments, q is an integer greater than or equal to 0. In certain embodiments, q is an integer greater than or equal to 1.

[0320] In some embodiments, e.g., where q is greater than 2, Aqis a group which is connected to a compound of formula (I), (IA), (IB), (IC), or (II), or a pharmaceutically acceptable salt, tautomer, or solvate thereof, target protein ligand, or combination thereof,and Ai and Aqare connected via structural units of A (number of such structural units of A: q-2).

[0321] In additional embodiments, q is an integer from 1 to 100, 1 to 90, 1 to 80, 1 to 70, 1 to 60, 1 to 50, 1 to 40, 1 to 30, 1 to 20, or 1 to 10.

[0322] Examples of linkers for PROTACs are well known in the art and can include those described in WO2017197055, W02017007612, WO2015160845, W02021077010, W02018071606, WO2020211822, W02020198711, W02020160295, and WO2020214952, each of which is herein incorporated by reference in its entirety. In addition, a wide range of linkers suitable for PROTAC development are commercially available from vendors including Selleck Chemicals, BroadPharm and MedChemExpress.com.

[0323] In some embodiments, the linker includes a group represented by the following structures:OwhereinWL1and WL2are each independently a 4-8 membered ring with 0-4 heteroatoms, optionally substituted with RQ, each RQ is independently a H, halo, OH, CN, CFj, Ci-C6alkyl (linear, branched, optionally substituted), Ci-Ce alkoxy (linear, branched, optionally substituted), or 2 RQ groups taken together with the atom they are attached to, form a 4-8 membered ring system containing 0-4 heteroatoms;YL1is each independently a bond, Ci-Ce alkyl (linear, branched, optionally substituted) and optionally one or more C atoms are replaced with O; or Ci-Ce alkoxy (linear, branched, optionally substituted);n and m are each independently an integer from 1 to 100, 1 to 90, 1 to 80, 1 to 70, 1 to 60, 1 to 50, 1 to 40, 1 to 30, 1 to 20, or 1 to 10; anda dashed line indicates the attachment point to the TRIM21 targeting compounds of formula (I), (IA), (IB), (IC), or (II) or a pharmaceutically acceptable salt, tautomer, or solvate thereof or target protein ligand moieties.

[0324] In additional embodiments, the linker group is optionally substituted polyethylene glycol having between 1 and about 100 ethylene glycol units, between about 1 and about 50 ethylene glycol units, between 1 and about 25 ethylene glycol units, between about 1 and 10 ethylene glycol units, between 1 and about 8 ethylene glycol units and 1 and 6 ethylene glycol units, between 2 and 4 ethylene glycol units, or optionally substituted alkyl groups interdispersed with optionally substituted, O, N, S, P or Si atoms. In certain embodiments, the linker is substituted with an aryl, phenyl, benzyl, alkyl, alkylene, or heterocycle group. In certain embodiments, the linker may be asymmetric or symmetrical.

[0325] In any of the embodiments of the compounds described herein, the linker group may be any suitable moiety as described herein. In one embodiment, the linker is a substituted or unsubstituted polyethylene glycol group ranging in size from about 1 to about 12 ethylene glycol units, between 1 and about 10 ethylene glycol units, about 2 about 6 ethylene glycol units, between about 2 and 5 ethylene glycol units, between about 2 and 4 ethylene glycol units.

[0326] Although a compound of formula (I), (IA), (IB), (IC), or (II), or a pharmaceutically acceptable salt, tautomer, or solvate thereof, and target protein ligand may be covalently linked to the linker group through any group which is appropriate and stable to the chemistry of the linker, in some embodiments, the linker is independently covalently bonded to the a compound of formula (I), (IA), (IB), (IC), or (II), or a pharmaceutically acceptable salt, tautomer, or solvate thereof, and the target protein ligand preferably through an amide, ester, thioester, keto group, carbamate (urethane), carbon or ether, each of which groups may be inserted anywhere on the compound of formula (I), (IA), (IB), (IC), or (II), or a pharmaceutically acceptable salt, tautomer, or solvate thereof, and target protein ligand to provide maximum binding of the compound of formula (I), (IA), (IB), (IC), or (II), or a pharmaceutically acceptable salt, tautomer, or solvate thereof, on the ubiquitin ligase TRIM21 and the target protein ligand on the target protein to be degraded. In certain preferred aspects, the linker may be linked to an optionally substituted alkyl, alkylene, alkeneor alkyne group, an aryl group or a heterocyclic group on the compound of formula (I), (IA), (IB), (IC), or (II), or a pharmaceutically acceptable salt, tautomer, or solvate thereof or the target protein ligand.

[0327] The target protein ligand is a group that binds to target proteins. Targets of the target protein ligand are numerous in kind and are selected from proteins that are expressed in a cell, such that at least a portion of the target protein is found in the cell and may bind to a target protein ligand. The term “protein” includes oligopeptides and polypeptide sequences of sufficient length that they can bind to a target protein ligand described herein. Any protein in a eukaryotic system or a microbial system, including a virus, bacteria or fungus, as otherwise described herein, are targets for ubiquitination mediated by the compounds and PROTACs described herein. Preferably, the target protein is an eukaryotic protein.

[0328] Target protein ligands can include, for example, any moiety that binds to a protein specifically (binds to a target protein). Non-limiting examples of small molecule target protein ligands include Hsp90 inhibitors, kinase inhibitors, HDM2 & MDM2 inhibitors, compounds targeting Human BET Bromodomain-containing proteins, HDAC inhibitors, human lysine methyltransferase inhibitors, angiogenesis inhibitors, nuclear hormone receptor compounds, immunosuppressive compounds, and compounds targeting the aryl hydrocarbon receptor (AHR), cancer diagnostic agents, and tau diagnostic agents, among numerous others.

[0329] Such small molecule target protein ligands can include pharmaceutically acceptable salts, enantiomers, solvates and polymorphs of these compositions, as well as other small molecules that may target a protein of interest. These binding moieties are linked to the compound of formula (I), (IA), (IB), (IC), or (II), or a pharmaceutically acceptable salt, tautomer, or solvate thereof preferably through a linker in order to present a target protein (to which the protein target moiety is bound) in proximity to TRIM21 for ubiquitination and degradation.

[0330] Any protein that can bind to a target protein ligand and be acted on or degraded by TRIM21 is a target protein according to embodiments described herein. Target proteins include proteins and peptides having any biological function or activity, including structural, regulatory, hormonal, enzymatic, genetic, immunological, contractile, storage, transportation, and signal transduction. In certain embodiments, the target proteins include structural proteins, receptors, enzymes, cell surface proteins, proteins pertinent to the integratedfunction of a cell, including proteins involved in catalytic activity, aromatase activity, motor activity, helicase activity, metabolic processes (anabolism and catabolism), antioxidant activity, proteolysis, biosynthesis, proteins with kinase activity, oxidoreductase activity, transferase activity, hydrolase activity, lyase activity, isomerase activity, ligase activity, enzyme regulator activity, signal transducer activity, structural molecule activity, binding activity (protein, lipid carbohydrate), receptor activity, cell motility, membrane fusion, cell communication, regulation of biological processes, development, cell differentiation, response to stimulus, behavioral proteins, cell adhesion proteins, proteins involved in cell death, proteins involved in transport (including protein transporter activity, nuclear transport, ion transporter activity, channel transporter activity, carrier activity, permease activity, secretion activity, or electron transporter activity), pathogenesis, chaperone regulator activity, nucleic acid binding activity, transcription regulator activity, extracellular organization, biogenesis activity, and translation regulator activity. Proteins of interest can include proteins from eukaryotes and prokaryotes, including microbes, viruses, fungi and parasites, including humans, microbes, viruses, fungi and parasites, among numerous others, as targets for drug therapy, other animals, including domesticated animals, microbials for the determination of targets for antibiotics, and other antimicrobials and plants, and even viruses, among numerous others.

[0331] More specifically, a number of drug targets for human therapeutics represent protein targets to which the target protein ligand may be bound and incorporated into PROTACs as described herein. These include protein targets which may be used to restore function in numerous polygenic diseases, including for example B7.1 and B7, TINFRlm, TNFR2, NADPH oxidase, BclIBax and other partners in the apoptosis pathway, C5a receptor, HMG-CoA reductase, PDE V phosphodiesterase type, PDE IV phosphodiesterase type 4, PDE I, PDEII, PDEIII, squalene cyclase inhibitor, CXCR1, CXCR2, nitric oxide (NO) synthase, cyclo-oxygenase 1, cyclo-oxygenase 2, alpha-synuclein, HIV capsid protein, VP40, BET-bromodomain protein, condensate-forming oncogenic fusion proteins including nucleoporin-containing fusion proteins (e.g., NIJP98-NSD1, NUP98-KDM5A, NUP98-PHF23; DEK-NUP214, NUP214-ABL1, SET-NUP214, etc.), EML4-ALK, and BRD4-NUT, or oligomeric proteins involved in inflammatory signaling including NLRP3, RIPK3, RIPK1, MLKL, and GSDM3, 5HT receptors, dopamine receptors, G Proteins, Gq, histamine receptors, 5 -lipoxygenase, tryptase serine protease, thymidylate synthase, purine nucleosidephosphorylase, GAPDH trypanosoma!, glycogen phosphorylase, Carbonic anhydrase, chemokine receptors, JAW STAT, RXR and similar, HIV 1 protease, HIV 1 integrase, influenza, neuramimidase, hepatitis B reverse transcriptase, sodium channel, multi drug resistance (MDR), protein P-glycoprotein (and MRP), tyrosine kinases, CD23, CD124, tyrosine kinase p56 lek, CD4, CD5, IL-2 receptor, IL-I receptor, TNF-aR, ICAM1, Cat+ channels, VCAM, VLA-4 integrin, selectins, CD40 / CD40L, newokinins and receptors, inosine monophosphate dehydrogenase, p38 MAP Kinase, Ras / Raf / ME / ERK pathway, interleukin- 1 converting enzyme, caspase, HCV, NS3 protease, HCV NS3 RNA helicase, glycinamide ribonucleotide formyl transferase, rhinovirus 3C protease, herpes simplex virus-1 (HSV-I), protease, cytomegalovirus (CMV) protease, poly (ADP-ribose) polymerase, cyclin dependent kinases, vascular endothelial growth factor, c-Kit, TGF0 activated kinase 1, mammalian target of rapamycin, SHP2, androgen receptor, oxytocin receptor, microsomal transfer protein inhibitor, bile acid transport inhibitor, 5 alpha reductase inhibitors, angiotensin 11, glycine receptor, noradrenaline reuptake receptor, estrogen receptor, estrogen related receptors, focal adhesion kinase, Src, endothelin receptors, neuropeptide Y and receptor, adenosine receptors, adenosine kinase and AMP deaminase, purinergic receptors (P2Y1, P2Y2, P2Y4, P2Y6, P2X1-7), famesyltransferases, geranylgeranyl transferase, TrkA a receptor for NGF, amyloid, tau or pathological tau aggregates, tyrosine kinase Flk-IIKDR, vitronectin receptor, integrin receptor, Her-21 neu, telomerase inhibition, cytosolic phospholipaseA2, or EGF receptor tyrosine kinase.

[0332] In other embodiments, proteins that can be targeted by the target protein ligand include CRBN, VHL, Tau, DHODH, FKBP12, AR, ERa, RAR, CRABP-II, ALK, CK2, GDK8 and GDK9, BTK, PI3K, TBK1, FLT3, BTK, RTKs such as EGFR, HER2 and cMET, ERK1 and ERK2, BCR-ABL, RIPK2, BCL6, PCAF / GCN5, BRD4 and HDAC6, TRIM24, VP40, SIRT2, BRD9, disclosed, example, in Scheepstra, M., Comput. Struct. Biotec. 17 (2019) 160-176; US 2018 / 0125821; US 2015 / 0291562; US 2017 / 0065719 Al, whrch are herein incorporated by reference in their entirety.

[0333] In still other embodiments, the proteins that can be targeted by the target protein ligand include tau, amyloid, alpha-synuclein, HIV capsid protein, VP40, condensate-forming oncogenic fusion proteins including nucleoporin-containing fusion proteins (e.g., NUP98-NSD1, NUP98-KDM5A, NUP98-PHF23; DEK-NUP214, NUP214-ABL1, SET-NUP214,etc.) EML4-ALK, and BRD4-NUT, or oligomeric proteins involved in inflammatory signaling including NLRP3, RIPK3, RIPK1, MLKL, and GSDM3.

[0334] In one example, the target protein ligand can include JQ-1, a ligand frequently used in PROTACs to promote degradation of BET bromodomain proteins, such as bromodomain-containing protein 4 (BRD4). Examples of PROTACs including a compound of formula (I) linked to JQ-1 can have a structure selected from:acceptable salt, tautomer, or solvate thereof.

[0335] In another example, the target protein ligand can include a pathological tau targeting compound, such as a clinical PET tracer for tauopathy diagnosis with high affinity and selectivity for pathological isoforms. Examples of PROTACs including a compound of formula (I) linked to the pathological tau targeting compound can have a structure selectedtautomer, or solvate thereof.

[0336] In some embodiments, the compounds, molecular glues, or PROTACs described herein may be formulated in a conventional manner with one or more pharmaceutically acceptable carriers, additives, diluents, or excipients to provide a pharmaceutical composition. Pharmaceutically acceptable carriers that may be used in these pharmaceutical compositions include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as prolamine sulfate,disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat.

[0337] The compositions as described herein may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. The term “parenteral” as used herein includes subcutaneous, intravenous, intramuscular, intra-articular, intra-synovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. Preferably, the compositions are administered orally, intraperitoneally or intravenously.

[0338] Sterile injectable forms of the compositions as described herein may be an aqueous or oleaginous suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example as a solution in 1,3 -butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer’s solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono- or di -glycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant.

[0339] The pharmaceutical compositions as described herein may be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, aqueous suspensions or solutions. In the case of tablets for oral use, carriers which are commonly used include lactose and com starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried corn starch. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring or coloring agents may also be added.

[0340] Alternatively, the pharmaceutical compositions as described herein may be administered in the form of suppositories for rectal administration. These can be prepared by mixing the agent with a suitable non-irritating excipient, which is solid at room temperature but liquid at rectal temperature and therefore will melt in the rectum to release the drug. Such materials include cocoa butter, beeswax and polyethylene glycols.

[0341] The pharmaceutical compositions as described herein may also be administered topically. Suitable topical formulations are readily prepared for each of these areas or organs. Topical application for the lower intestinal tract can be effected in a rectal suppository formulation or in a suitable enema formulation. Topically-acceptable transdermal patches may also be used.

[0342] For topical applications, the pharmaceutical compositions may be formulated in a suitable ointment containing the active component suspended or dissolved in one or more carriers. Carriers for topical administration of the compounds of this invention include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water. In certain preferred aspects of the invention, the compounds may be coated onto a stent which is to be surgically implanted into a patient in order to inhibit or reduce the likelihood of occlusion occurring in the stent in the patient.

[0343] Alternatively, the pharmaceutical compositions can be formulated in a suitable lotion or cream containing the active components suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

[0344] For ophthalmic use, the pharmaceutical compositions may be formulated as micronized suspensions in isotonic, pH adjusted sterile saline, or, preferably, as solutions in isotonic, pH adjusted sterile saline, either with our without a preservative such as benzylalkonium chloride. Alternatively, for ophthalmic uses, the pharmaceutical compositions may be formulated in an ointment such as petrolatum.

[0345] The pharmaceutical compositions as described herein may also be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline,employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and / or other conventional solubilizing or dispersing agents.

[0346] The amount of compound, molecular glue, or PROTAC in a pharmaceutical composition as described herein that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host and disease treated, the particular mode of administration. It should also be understood that a specific dosage and treatment regimen for any particular patient will depend upon a variety of factors, including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the judgment of the treating physician and the severity of the particular disease or condition being treated.

[0347] A patient or subject in need of therapy using the compounds, molecular glues, or PROTACs described herein according to the methods described herein can be treated by administering to the patient (subject) an effective amount of the compound according to the present invention including pharmaceutically acceptable salts, solvates or polymorphs, thereof optionally in a pharmaceutically acceptable carrier or diluent, either alone, or in combination with other bioactive agents.

[0348] The active compounds, molecular glues, or PROTACs described herein are included in the pharmaceutically acceptable earner or diluent in an amount sufficient to deliver to a patient a therapeutically effective amount for the desired indication, without causing serious toxic effects in the patient treated. Exemplary doses can be from about 0.01 to about 1000 mg, by oral administration. Examples of dose ranges can include from a minimum dose of about 0.01, 0.10, 0.50, 1, 5, 10, 25, 50, 100, 125, 150, 200, or 250 mg to a maximum dose of about 300, 400, 500, 600, 700, 800, 900, or 1000 mg, wherein the dose range can include from any one of the foregoing minimum doses to any one of the foregoing maximum doses. Specific examples of particular effective amounts contemplated via oral administration can include about 0.02, 0.03, 0.04, 0.05, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.85. 0.90, 0.95, 1, 2, 3, 4. 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99. 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 205, 210, 215, 220, 225,230, 235, 240, 245, 250, 255, 260, 265, 270, 275, 280, 285, 290, 295, 300, 305, 310, 315, 320, 325, 330, 335, 340, 345, 350, 355, 360, 365, 370, 375, 380, 385, 390, 395, 400, 405, 410, 415, 420, 425, 430, 435, 440, 445, 450, 455, 460, 465, 470, 475, 480, 485, 490, 495, 500, 505, 510, 515, 520, 525, 530, 535, 540, 545, 550, 555, 560, 565, 570, 575, 580, 585, 590, 595, 600, 605, 610, 615, 620, 625, 630, 635, 640, 645. 650, 655, 660, 665, 670, 675, 680, 685, 690, 695, 700, 705, 710, 715, 720, 725, 730, 735, 740, 745, 750, 755, 760, 765, 770, 775, 780, 785, 790, 795, 800, 805, 810, 820, 825, 830, 835, 840, 845, 850, 855, 860, 865, 870, 875, 880, 885, 890, 895, 900, 905, 910, 915, 920, 925, 930, 935, 940, 945, 950, 955, 960, 965, 970, 975, 980, 985, 990, 995, 1000 mg or more. The oral dose can be administered once daily, twice daily, three times daily, or more frequently.

[0349] The compounds, molecular glues, or PROTACs described herein for use in parenteral administration (e.g., intravenous administration) is generally from about 0.01 to about 300 mg / kg body weight. Examples of dose ranges can include from a minimum dose of about 0.01, 0.10, 0.50, 1, 5, 10, 25, 50, or 100 mg / kg body weight to a maximum dose of about 125, 150, 175, 200, 250, 275, or 300 mg / kg body weight, wherein the dose range can include from any one of the foregoing minimum doses to any one of the foregoing maximum doses. Specific examples of effective amounts contemplated include about 0.02, 0.03, 0.04, 0.05, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50. 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, 270, 275, 280, 285, 290, 295, 300 mg / kg body weight or more. Continuous intravenous administration is also contemplated for from 1 to 24 hours per day to achieve a target concentration from about 0.01 mg / L blood to about 100 mg / L blood. Exemplary dose ranges can include from a minimum dose of about 0.01, 0.10, 0.25, 0.50, 1, 5, 10, or 25 mg / L blood to a maximum dose of about 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, or 100 mg / L, wherein an exemplary dose ranges can include from any one of the foregoing minimum doses to any one of the foregoing maximum doses. Specific examples of particular effective amounts contemplated via this route include about 0.02, 0.03, 0.04, 0.05, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12. 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27. 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58,59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100 mg / L blood or more. The dose to be used can depend upon various conditions, and there may be cases wherein doses lower than or greater than the ranges specified above are used.

[0350] The compounds, molecular glues, or PROTACs described herein are preferably administered to achieve peak plasma concentrations of the active compound of about 0.00001-30 mM, preferably about 0.1-30 pM. This may be achieved, for example, by the intravenous injection of a solution or formulation of the active ingredient, optionally in saline, or an aqueous medium or administered as a bolus of the active ingredient. Oral administration is also appropriate to generate effective plasma concentrations of the active agent.

[0351] The concentration of the compounds, molecular glues, or PROTACs described herein in the composition will depend on absorption, distribution, inactivation, and excretion rates of the compounds, molecular glues, or PROTACs described herein as well as other factors known to those of skill in the art. It is to be noted that dosage values will also vary with the severity of the condition to be alleviated. It is to be further understood that for any particular subject, specific dosage regimens should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions, and that the concentration ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed composition. The active ingredient may be administered at once, or may be divided into a number of smaller doses to be administered at varying intervals of time.

[0352] The compounds, molecular glues, or PROTACs described herein can also be mixed with other active materials that do not impair the desired action, or with materials that supplement the desired action. In certain preferred aspects, one or more compounds, molecular glues, or PROTACs described herein can be co-administered with another bioactive agent.

[0353] Solutions or suspensions used for parenteral, intradermal, subcutaneous, or topical application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such asethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. The parental preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.

[0354] If administered intravenously, preferred carriers are physiological saline or phosphate buffered saline (PBS).

[0355] In one embodiment, the compounds, molecular glues, or PROTACs described herein are prepared with carriers that will protect the compound against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, poly anhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Methods for preparation of such formulations will be apparent to those skilled in the art.

[0356] Liposomal suspensions may also be pharmaceutically acceptable carriers. These may be prepared according to methods known to those skilled in the art, for example, as described in U. S. Pat. No. 4,522,811 (which is incorporated herein by reference in its entirety). For example, liposome formulations may be prepared by dissolving appropriate lipid(s) (such as stearoyl phosphatidyl ethanolamine, stearoyl phosphatidyl choline, arachadoyl phosphatidyl choline, and cholesterol) in an inorganic solvent that is then evaporated, leaving behind a thin film of dried lipid on the surface of the container. An aqueous solution of the active compound are then introduced into the container. The container is then swirled by hand to free lipid material from the sides of the container and to disperse lipid aggregates, thereby forming the liposomal suspension.

[0357] In some embodiments, therapeutic compositions can include an effective amount of compounds, molecular glues, or PROTACs described herein, and a pharmaceutically acceptable carrier. The therapeutic compositions can modulate protein degradation in a patient or subject, for example, an animal such as a human, and can be used for treating or ameliorating disease states or conditions that are modulated through the degraded protein.

[0358] The terms “treat”, “treating”, and “treatment”, etc., as used herein, refer to any action providing a benefit to a patient for which the present compounds, molecular glues, or PROTACs may be administered, including the treatment of any disease state or conditionthat is modulated through the protein to which the present compounds, molecular glues, or PROTACs bind.

[0359] In some embodiments, therapeutic compositions, including the compounds, molecular glues, or PROTACs as described herein, can be used for effectuating the degradation of proteins of interest and the treatment or amelioration of a disease, e.g., cancer. As such, the compounds, molecular glues, or PROTACs as described herein can be used in a method of ubiquitinating / degrading a target protein in a cell.

[0360] In certain embodiments, the method includes administering compounds, molecular glues, or PROTACs described herein that include a compound of formula (I), (IA), (IB), (IC), or (II), or a pharmaceutically acceptable salt, tautomer, or solvate thereof linked to a target protein ligand wherein the compound of formula (I), (IA), (IB), (IC), or (II), or a pharmaceutically acceptable salt, tautomer, or solvate thereof recognizes TRIM21 and the target protein ligand recognizes the target protein such that degradation of the target protein will occur when the target protein is placed in proximity to the TRIM21, thus resulting in degradation / inhibition of the effects of the target protein and the control of protein levels. The control of protein levels afforded by the methods described herein provides treatment of a disease state or condition, which is modulated through the target protein by lowering the level of that protein in the cell, e.g., cell of a patient. In certain embodiments, the methods comprise administering an effective amount of a compound, molecular glue, or PROTAC as described herein, optionally including a pharmaceutically acceptable excipient, carrier, adjuvant, another bioactive agent or combination thereof.

[0361] Other embodiments described herein relate to methods for treating or ameliorating a disease, disorder or symptom thereof in a subject or a patient, e.g., an animal such as a human, by administering to the subject in need thereof a composition comprising an effective amount, e.g., a therapeutically effective amount, of a compound, molecular glue, or PROTAC as described herein, and a pharmaceutically acceptable excipient, carrier, adjuvant, another bioactive agent or combination thereof, wherein the composition is effective for treating or ameliorating the disease or disorder or symptom thereof in the subject.

[0362] Still other embodiments described herein relate to methods for identifying the effects of the degradation of proteins of interest in a biological system using a compound, molecular glue, or PROTAC as described herein.

[0363] Other embodiments described herein relate to a method of treating a human patient in need thereof for a disease state or condition modulated through a protein where the degradation of that protein will produce a therapeutic effect in that patient. The method can include administering to a patient in need an effective amount of a compound, molecular glue, or PROTAC as described herein, optionally in combination with another bioactive agent. The disease state or condition may be a disease caused by a microbial agent or other exogenous agent, such as a virus, bacteria, fungus, protozoa or other microbe or may be a disease state, which is caused by overexpression of a protein, which leads to a disease state and / or condition

[0364] The term “disease state or condition” is used to describe any disease state or condition wherein protein dysregulation (i.e., the amount of protein expressed in a patient is elevated) occurs and where degradation of one or more proteins in a patient may provide beneficial therapy or relief of symptoms to a patient in need thereof. In certain instances, the disease state or condition may be cured.

[0365] Disease states of conditions which may be treated using compounds, molecular glues, or PROTACs described herein include, for example, asthma, autoimmune diseases, such as multiple sclerosis, various cancers, ciliopathies, cleft palate, diabetes, heart disease, hypertension, inflammatory bowel disease, mental retardation, mood disorder, obesity, refractive error, infertility, Angelman syndrome, Canavan disease, Coeliac disease, Charcot-Marie-Tooth disease, Cystic fibrosis, Duchenne muscular dystrophy, Haemochromatosis, Haemophilia, Klinefelter's syndrome, Neurofibromatosis, Phenylketonuria, Polycystic kidney disease, (PKD1) or 4 (PKD2) Prader-Willi syndrome, Sickle-cell disease, Tay-Sachs disease, Turner syndrome.

[0366] Further disease states or conditions, which may be treated using compounds, molecular glues, or PROTACs described herein include Alzheimer’s disease, Amyotrophic lateral sclerosis (Lou Gehrig's disease), Anorexia nervosa, Anxiety disorder, Atherosclerosis, Attention deficit hyperactivity disorder, Autism, Bipolar disorder, Chronic fatigue syndrome, Chronic obstructive pulmonary disease, Crohn's disease, Coronary heart disease, Dementia, Depression, Diabetes mellitus type 1, Diabetes mellitus type 2, Epilepsy, Guillain-Barre syndrome, Irritable bowel syndrome, Lupus, Metabolic syndrome, Multiple sclerosis, Myocardial infarction, Obesity, Obsessive-compulsive disorder, Panic disorder, Parkinson'sdisease. Psoriasis, Rheumatoid arthritis, Sarcoidosis, Schizophrenia, Stroke, Thromboangiitis obliterans, Tourette syndrome, Vasculitis.

[0367] Still additional disease states or conditions which can be treated by using compounds, molecular glues, or PROTACs as described herein include aceruloplasminemia, Achondrogenesis type II, achondroplasia, Acrocephaly, Gaucher disease type 2, acute intermittent porphyria, Canavan disease, Adenomatous Polyposis Coli, ALA dehydratase deficiency, adenylosuccinate lyase deficiency, Adrenogenital syndrome, Adrenoleukodystrophy, ALA-D porphyria, ALA dehydratase deficiency, Alkaptonuria, Alexander disease, Alkaptonuric ochronosis, alpha 1 -antitrypsin deficiency, alpha- 1 proteinase inhibitor, emphysema, amyotrophic lateral sclerosis Alstrom syndrome, Alexander disease, Amelogenesis imperfecta, ALA dehydratase deficiency, Anderson-Fabry disease, androgen insensitivity syndrome, Anemia Angiokeratoma Corporis Diffusum, Angiomatosis retinae (von Hippel-Lindau disease) Apert syndrome, Arachnodactyly (Marfan syndrome), Stickler syndrome, Arthrochalasis multiplex congenital (Ehlers-Danlos syndrome arthrochalasia type) ataxia telangiectasia, Rett syndrome, primary pulmonary hypertension, Sandhoff disease, neurofibromatosis type II, Beare-Stevenson cutis gyrata syndrome, Mediterranean fever, familial, Benjamin syndrome, beta-thalassemia, Bilateral Acoustic Neurofibromatosis (neurofibromatosis type II), factor V Leiden thrombophilia, Bloch-Sulzberger syndrome (incontinentia pigmenti), Bloom syndrome, X-linked sideroblastic anemia, Bonnevie-Ullrich syndrome (Turner syndrome), Bourneville disease (tuberous sclerosis), prion disease, Birt-IIogg-Dube syndrome, Brittle bone disease (osteogenesis imperfecta), Broad Thumb-Hallux syndrome (Rubinstein-Taybi syndrome), Bronze Diabetes / Bronzed Cirrhosis (hemochromatosis), Bulbospinal muscular atrophy (Kennedy's disease), Burger-Grutz syndrome (lipoprotein lipase deficiency), CGD Chronic granulomatous disorder, Campomelic dysplasia, biotinidase deficiency, cancer Cardiomyopathy (Noonan syndrome), Cri du chat, CAVD (congenital absence of the vas deferens), Caylor cardiofacial syndrome (CBAVD), CEP (congenitalerythropoietic porphyria), cystic fibrosis, congenital hypothyroidism, Chondrodystrophy syndrome (achondroplasia), otospondylomegaepiphyseal dysplasia, Lesch-Nyhan syndrome, galactosemia, Ehlers-Danlos syndrome, Thanatophoric dysplasia, Coffin-Lowry syndrome, Cockayne syndrome, (familial adenomatous polyposis), Congenital erythropoietic porphyria, Congenital heart disease, Methemoglobinemia / Congenital methaemoglobinaemia,achondroplasia, X-linked sideroblastic anemia, Connective tissue disease, Conotruncal anomaly face syndrome, Cooley’s Anemia (beta-thalassemia), Copper storage disease (Wilson's disease), Copper transport disease (Menkes disease), hereditary coproporphyria, Cowden syndrome, Craniofacial dysarthrosis (Crouzon syndrome), Creutzfeldt- Jakob disease (prion disease), Cockayne syndrome, Cowden syndrome, Curschmann-Batten-Steinert syndrome (myotonic dystrophy), Beare-Stevenson cutis gyrata syndrome, primary hyperoxaluria, spondyloepimetaphyseal dysplasia (Strudwick type), muscular dystrophy, Duchenne and Becker types (DBMD), Usher syndrome. Degenerative nerve diseases including de Grouchy syndrome and Dejerine-Sottas syndrome, developmental disabilities, distal spinal muscular atrophy, type V, androgen insensitivity syndrome, Diffuse Globoid Body Sclerosis (Krabbe disease), Di George's syndrome, Dihydrotestosterone receptor deficiency, androgen insensitivity syndrome, Down syndrome, Dwarfism, erythropoietic protoporphyria Erythroid 5 -aminolevulinate synthetase deficiency, Erythropoietic porphyria, erythropoietic protoporphyria, erythropoietic uroporphyria, Eriedreich's ataxia, familial paroxysmal polyserositis, porphyria cutanea tarda, familial pressure sensitive neuropathy, primary pulmonary hypertension (PPH), Fibrocystic disease of the pancreas, fragile X syndrome, galactosemia, genetic brain disorders, Giant cell hepatitis (Neonatal hemochromatosis), Gronblad-Strandberg syndrome (pseudoxanthoma elasticum), Gunther disease (congenital erythropoietic porphyria), haemochromatosis, Hallgren syndrome, sickle cell anemia, hemophilia, hepatoerythropoietic porphyria (HEP), Hippel-Lindau disease (von Ilippel-Lindau disease), Huntington's disease, Hutchinson-Gilford progeria syndrome (progeria), Hyperandrogenism, Hypochondroplasia, Hypochromic anemia, Immune system disorders, including X-linked severe combined immunodeficiency, Insley-Astley syndrome, Jackson-Weiss syndrome, Joubert syndrome, Lesch-Nyhan syndrome, Jackson-Weiss syndrome, Kidney diseases, including hyperoxaluria, Klinefelter's syndrome, Kniest dysplasia, Lacunar dementia, Langer-Saldino achondrogenesis, ataxia telangiectasia, Lynch syndrome, LysyLhydroxylase deficiency, Machado-Joseph disease, Metabolic disorders, including Kniest dysplasia, Marfan syndrome, Movement disorders, Mowat-Wilson syndrome, cystic fibrosis, Muenke syndrome, Multiple neurofibromatosis, Nance-Insley syndrome, Nance-Sweeney chondrodysplasia, neoplasia, Niemann-Pick disease, Noack syndrome (Pfeiffer syndrome), Osler- Weber- Rendu disease, Peutz-Jeghers syndrome, Polycystic kidney disease, polyostotic fibrous dysplasia (McCune- Albright syndrome),Peutz-Jeghers syndrome, Prader-Labhart-Willi syndrome, hemochromatosis, primary hyperuricemia syndrome (Lesch-Nyhan syndrome), primary pulmonary hypertension, primary senile degenerative dementia, prion disease, progeria (Hutchinson Gilford Progeria Syndrome), progressive chorea, chronic hereditary (Huntington) (Huntington's disease), progressive muscular atrophy, spinal muscular atrophy, propionic acidemia, protoporphyria, proximal myotonic dystrophy, pulmonary arterial hypertension, PXE (pseudoxanthoma elasticum), Rb (retinoblastoma), Recklinghausen disease (neurofibromatosis type I), Recurrent polyserositis, Retinal disorders, Retinoblastoma, Rett syndrome, REALS type 3, Ricker syndrome, Riley-Day syndrome, Roussy-Levy syndrome, severe achondroplasia with developmental delay and acanthosis nigricans (SADDAN), Li-Fraumeni syndrome, sarcoma, breast, leukemia, and adrenal gland (SBLA) syndrome, sclerosis tuberose (tuberous sclerosis), SDAT, SED congenital (spondyloepiphyseal dysplasia congenita), SED Strudwick (spondyloepimetaphyseal dysplasia, Strudwick type), SEDc (spondyloepiphyseal dysplasia congenita) SEMD, Strudwick type (spondyloepimetaphyseal dysplasia, Strudwick type), Shprintzen syndrome, Skin pigmentation disorders, Smith-Lemli-Opitz syndrome, South-African genetic porphyria (variegate porphyria), infantile-onset ascending hereditary spastic paralysis. Speech and communication disorders, sphingolipidosis, Tay-Sachs disease, spinocerebellar ataxia, Stickler syndrome, stroke, androgen insensitivity syndrome, tetrahydrobiopterin deficiency, beta-thalassemia, Thyroid disease, Tomaculous neuropathy (hereditary neuropathy with liability to pressure palsies), Treacher Collins syndrome, Triple X syndrome (triple X syndrome), Trisomy 21 (Down syndrome), Trisomy X, VHL syndrome (von Hippel-Lindau disease), Vision impairment and blindness (Alstrbm syndrome), Vrolik disease, Waardenburg syndrome, Warburg Sjo Fledelius Syndrome, Weissenbacher-Zweymiiller syndrome, Wolf-Hirschhorn syndrome, Wolff Periodic disease, Weissenbacher-Zweymiiller syndrome and Xcruderma pigmentosum, among others.

[0368] The term “neoplasia” or “cancer” is used throughout the specification to refer to the pathological process that results in the formation and growth of a cancerous or malignant neoplasm, i.e., abnormal tissue that grows by cellular proliferation, often more rapidly than normal and continues to grow after the stimuli that initiated the new growth cease.Malignant neoplasms show partial or complete lack of structural organization and functional coordination with the normal tissue and most invade surrounding tissues, metastasize to several sites, and are likely to recur after attempted removal and to cause the death of thepatient unless adequately treated. As used herein, the term neoplasia is used to describe all cancerous disease states and embraces or encompasses the pathological process associated with malignant hematogenous, ascitic and solid tumors. Exemplary cancers which may be treated by the present compounds, molecular glues, or PROTACs either alone or in combination with at least one additional anti-cancer agent include squamous-cell carcinoma, basal cell carcinoma, adenocarcinoma, hepatocellular carcinomas, and renal cell carcinomas, cancer of the bladder, bowel, breast, cervix, colon, esophagus, head, kidney, liver, lung, neck, ovary, pancreas, prostate, and stomach; leukemias; benign and malignant lymphomas, particularly Burkitt's lymphoma and Non-Hodgkin's lymphoma; benign and malignant melanomas; myeloproliferative diseases; sarcomas, including Ewing's sarcoma, hemangiosarcoma, Kaposi’s sarcoma, liposarcoma, myosarcomas, peripheral neuroepithelioma, synovial sarcoma, gliomas, astrocytomas, oligodendrogliomas, ependymomas, gliobastomas, neuroblastomas, ganglioneuromas, gangliogliomas, medulloblastomas, pineal cell tumors, meningiomas, meningeal sarcomas, neurofibromas, and Schwannomas; bowel cancer, breast cancer, prostate cancer, cervical cancer, uterine cancer, lung cancer, ovarian cancer, testicular cancer, thyroid cancer, astrocytoma, esophageal cancer, pancreatic cancer, stomach cancer, liver cancer, colon cancer, melanoma; carcinosarcoma, Hodgkin's disease, Wilms' tumor and teratocarcinomas. Additional cancers which may be treated using compounds, molecular glues, or PROTACs describe herein include, for example, T-lineage Acute lymphoblastic Leukemia (T-ALL), T-lineage lymphoblastic Lymphoma (T-LL), Peripheral T-cell lymphoma, Adult T-cell Leukemia, Pre-B ALL, Pre-B Lymphomas, Large B-cell Lymphoma, Burkitts Lymphoma, B-cell ALL, Philadelphia chromosome positive ALL and Philadelphia chromosome positive CML.

[0369] In some embodiments, a compound, molecular glue, or PROTAC as described herein can be used for targeted protein degradation, such as pathological protein degradation or pathological aggregated protein degradation, such as for targeted degradation of pathogenic tau.

[0370] In some embodiments, a method for regulating protein activity of tau protein by degenerating tau aggregates in a patient in need thereof includes administering to the patient an amount of a compound, molecular glue, or PROTAC that binds to tau, preferably aggregated or pathological tau.

[0371] In still additional embodiments, a method of treating a disease state or condition in a patient wherein dysregulated protein activity (tau aggregation and accumulation) is responsible for the disease state or condition can include administering to said patient an effective amount of a compound, molecular glue, or PROTAC as described herein to the patient in order to regulate said protein activity in said patient. In certain embodiments, the protein is tau.

[0372] In another aspect, the disclosure provides methods of modulating tau protein ubiquitination and degradation in a subject, e.g., a cell, a tissue, mammal, or human patient. The method includes administering an effective amount of a compound, molecular glue, or PROTAC that binds to tau as described herein to a subject, wherein the compound, molecular glue, or PROTAC is effective in modulating tau protein ubquitination and degradation of the protein in the subject.

[0373] In another embodiment, a methods of treating or ameliorating a symptom of a disease related to tau accumulation or aggregation in a subject, e.g., a cell, a tissue, mammal, or human patient, can include administering an effective amount of a compound, molecular glue, or PROTAC that binds to tau as described herein, wherein the compound, molecular glue, or PROTAC as described herein is effective in treating or ameliorating a symptom of a disease related to tau aggregation in the subject.

[0374] Other embodiments relate to a compound described herein for use in aggregated protein degradation for central nervous system diseases, neurodegenerative diseases, and other diseases, such as Alzheimer’s disease and frontotemporal dementia.

[0375] The invention is further illustrated by the following example, which is not intended to limit the scope of the claims.Example 1

[0376] This example describes the identification and evaluation of TRIM21 -targeting small molecules and analogues thereof. We implemented a screening strategy in which pharmacological inhibition of all E3 ligases independently led to the identification of PRLX-93936, a cytotoxin of unknown mechanism of action, and BMS-214662, a famesyltransferase inhibitor, as molecular glue degraders targeting TRIM21. These molecules are structurally unrelated to hydroxy-acepromazine or HGC652 / HGClg, and newly-synthesized analogs of PRLX-93936 show substantially superior cellular potency. Like other known TRIM21 glues,we show that PRLX-93936 and BMS-214662 require TRIM21 activity for cytotoxicity and induce rapid degradation of a wide swath of nuclear pore proteins, leading to loss of nuclear trafficking and cell death. We also confirm that genetic disruption of NUP98’s autoproteolysis domain effectively prevents PRLX-93936-mediated nucleoporin degradation and cell death, further supporting a direct interaction between TRIM21 and NUP98.Evaluation of 30 PRLX-93936 analogs establishes the SAR of this series and enables the identification of a TRIM21 -targeting PROTAC, or TRIMT AC, that selectively degrades an engineered protein aggregate. Notably, in contrast to hydroxy-acepromazine-derived TRIMTACs, a TRIM21 -targeting PROTACs or TRIMTACs described herein do not require expression of a TRIM21 mutant for efficacy. Together, these studies define the anticancer mechanism of PRLX-93936 and BMS-214662 and provide high-quality glues, PROTACs, and TRIMTACs to propel the development of degraders of multimcric protein aggregates.MethodsStatistical analysis

[0377] All statistical analyses were performed in GraphPad Prism unless otherwise stated. Replicate description and statistical tests are described in the associated figure legends. Unless otherwise stated, normality was assumed for statistical tests that require this assumption (Pearson correlation, t-test). All statistical tests were two-tailed, and all P values are reported as exact values unless otherwise noted. No corrections for multiple comparisons were made.Cell lines

[0378] Cell lines were incubated at 37° C with 5% CO2 under humidified conditions. Lenti-X 293 T cells were purchased from Takara Bio. JUR-KAT E6-1 (TIB-152) and A549 were a gift of Dr. Mohamed Abazeed (Northwestern). OCT- AML-3 cells were a gift from Dr. David Wald (CWRU). C33a cells were a gift from Dr. John Pink (CWRU). HEK 293 T cells were a gift from Dr. Derek Taylor (CWRU). OCI-AML-3, JURKAT cell lines were maintained in glutamine-containing RPML1640 supplemented with 10% fetal bovine serum (FBS), and 1% penicillin / streptomycin. Lenti-X 293 T, HEK 293 T, and A549 cells were maintained in DMEM medium supplemented with 10% FBS, 2 mM L-glutamine, and 1%penicillin / streptomycin. C33a cells were maintained in glutamine-containing EMEM medium supplemented with 10% FBS and 1% penicillin / streptomycin.High-throughput chemical screen

[0379] OCI-AML-3 cells were seeded in a sterile tissue culture 384-well plate (Coming, 3765) at 1000 cells and 50 pl of media per well using an EL406 Microplate Washer Dispenser (Biotek). Screening library molecules (a collection of 1754 molecules derived from the Sigma LOPAC1280 and Selleck Bioactive Compound Library L1700 collections) were maintained as a 3 mM DMSO stock in Abgene storage 384-well plates (ThermoFisher Scientific; AB 1055). Transfer of 100 nL of each library molecule to assay plates was achieved using a solid pin tool, resulting in a final screening concentration of 6 pM. DMSO was used as a negative control within each plate. From each library plate, two replicate assay plates were generated that were identical except one plate was co-treated with 200 nM TAK-243 (added directly to cells and media 1 h before dispensing of library compounds). Plates were then incubated at 37°C for 10 h, at which time 5 pL of CellTiter-Glo reagent (Promega, G7572) was added to each well using the EL406 Microplate Washer Dispenser (Biotek). Plates were allowed to equilibrate for 10 min with shaking at room temperature before luminescence was immediately read using a Synergy Neo2 Multimode microplate reader (Biotek). Viability was calculated relative to DMSO wells within each plate, and hits were called on the basis of at least 50% reduction in viability in the DMSO conditions as well as a % viability gain of at least 30% in the TAK-243 condition.Cell viability assay

[0380] OCI-AML-3 and JURKAT cells were plated at a density of 1000 cells in 50 pl of culture medium in 384-well plates and treated with inhibitors same day before incubation at 37°C. For C33a, A549, and HEK 293 T cells, 500 cells in 50 pl in 384-well plates were used. After 24 h (OCI-AML3, JURKAT) or 72 h (C33a, A549, HEK 293 T), 5 pl of CellTiter-Glo (Promega) was added to each well, and the contents were mixed on an orbital shaker at room temperature for 10 min. Luminescence was recorded using a Biotek Synergy Nco2 microplatc reader with Gcn5 3.03.14 software. Dose curves were fitted using a four-parameter variable slope model in GraphPad Prism.Correlation analysis of cancer cell line profiling data

[0381] PRISM (23Q2) drug sensitivity and Expression Public (23Q2) gene expression data were retrieved from the Cancer Dependency Map (DepMap). PRISM (23 Q2) data are expressed as a log2 fold change for each cell line and Expression Public (23Q2) data are expressed as log2(Transcripts per Million+1). Pearson correlation coefficients and P values were calculated within the DepMap Custom Analysis tool by comparing drug sensitivity to gene expression profiles across cell lines with a minimum of 453 cell lines used for each correlation. Statistical tests associated with the Pearson correlation were two-sided and no correction for multiple comparisons was implemented.Lentivirus production and stable cell line generation

[0382] Lentiviral ORF-containing plasmids were synthesized and sequence validated by Vectorbuilder. Lenti-X 293T cells were plated in 6 well plates and allowed to attach overnight at 37°C. Cells were then co-transfected with lentiviral plasmids, psPAX2 (a gift from Didier Trono, Addgene #12260), and pMD2. G (a gift from Didier Trono, Addgene #12259) using Lipofectamine 2000 (Invitrogen, 11668027) per manufacturer’s instructions. After 6 h, media was replaced, and cells were incubated for 48 h. Eentivirus containing media was collected and filtered through a 0.45 pm filter before being directly added to cells with 8 pg / mL polybrene (Sigma Aldrich TR-1003-G). After 24 h at 37°C, media were replaced, and cells were incubated for another 24 h before selection with puromycin or blasticidin.

[0383] A549 PML-eGFP-BRD4(BD2) clones were generated by limiting dilution in 96-wcll plates and cultured over two weeks with a media change on day 7. Clones were then screened by fluorescence imaging on an Operetta High Content Imaging and Analysis System (Perkin Elmer) for intermediate PML-eGFP-BRD4(BD2) expression and consistent focus morphology. Selected clones were then transduced with lentivirus containing a TRIM21-FLAG allele.Western blotting

[0384] Cells were washed twice with phosphate buffered saline (PBS) and lysed in Pierce RIPA Buffer (Thermo Fisher Scientific, 89900) supplemented with IX cOmplete™ Mini EDTA-frcc Protease Inhibitor Cocktail (Roche, 11836170001). OCI-AML-3 cells werewashed twice with PBS and resuspended in PBS supplemented with IX cOmplete Mini EDTA-free Protease Inhibitor Cocktail before being lysed with two freeze-thaw cycles in liquid nitrogen. Lysates were clarified by centrifugation at 20,000g for 20 min at 4°C. Soluble protein concentrations were quantified with a Pierce BCA Assay (Thermo Fisher Scientific, 23225). Lysates were normalized by protein concentration and separated on 4- 12% or 3-8% gradient gels (Invitrogen, NW04125BOX or TA0381 BOX) before being transferred to PVDF membranes using the XCell II Blot Module (Thermo Fisher Scientific, EI9051). Membranes were blocked in 5% milk in TBST and incubated in primary antibody overnight at 4°C. Primary antibodies used: anti-P-actin (Sigma- Aldrich, A3854, 1:10,000), anti-TRIM21 (Proteintech, 12108-1-AP, 1:1000), anti-FLAG M2 (Sigma-Aldrich, F3165), anti-NUP214 (Abeam, ab70497, 1:500), anti-NUP88 (Santa Cruz Biotechnology, sc-365868), anti-NUP98 (Abclonal, A0530). Membranes were then incubated with secondary antibody conjugated to horseradish peroxidase (Cell Signaling Technology, 7076 or 7074, 1: 1000). Membranes were developed using either SuperSignal West Pico Plus Chemiluminescent Substrate (Thermo Fisher Scientific, 34580) or SuperSignal West Femto Maximum Sensitivity Substrate (Thermo Fisher Scientific, 34095) before imaging using a LL COR Odyssey Fc Imaging System.Liquid chromatography-tandem mass spectrometry

[0385] After incubation, cells were washed twice with PBS and resuspended in PBS supplemented with IX cOmplete™ Mini EDTA-free Protease Inhibitor Cocktail (Roche, 11836170001). Cells were lysed with a Fisher Scientific Sonic Dismembrator Model 60 using 15 x 1-s pulses at a power level of 3 at 4°C. Protein concentrations were quantified using a Pierce BCA Protein Assay kit (Thermo Fisher Scientific, 23225), and colorimetric development was measured using a Biotek Synergy Neo2 microplate reader. 50 pg of protein was denatured in 8 M Urea, reduced with 10 mM dithiothreitol, and alkylated with 25 mM iodoacetamide. Samples were then diluted with 100 mM Ammonium Bicarbonate and digested with 1 pg of Lys-C (Promega, VI 671) over-night at room temperature. Peptides were desalted on Cl 8 reverse-phase spin columns (BioPureSPN Mini FastEq, TARGA Cl 8, 120 A, The Nest Group, HUM S18R) before elution with 0.1% formic acid in 80% acetonitrile and 20% water. Peptides were dried and reconstituted in 0.1% formic acid in water for LC-MS / MS analysis.

[0386] Samples were analyzed by LC-MS using a timsTof Pro2 instrument (Bruker) equipped with a NanoElute UHPLC system. A 5 pl aliquot of each digest was injected onto a ThermoScientific (0.5 x 5 mm) Acclaim Pepmap Cis, 5-prn, trapping column. Liquid chromatographic elution was performed using a flow rale of 0.3 pl / min on a reverse phase column (ReproSil AQ C18, 75 pmx 150 mm, 1.9-pm 120-A). Peptide elution was performed using a binary gradient of mobile phase A, 0.1 % formic acid and mobile phase B, 0.1 % formic acid in acetonitrile. Each sample was analyzed using a linear gradient starting at 2% at 0 minutes and increasing to 35% B in 50 min, followed by an increase to 90% B in 2 min, then holding at 90% for 5 min before re-equilibration at 2% B. The electrospray voltage was 1500 V. A PASEF-DDA method was utilized for peptide identification. MSI scans were carried out with a resolution of 30,000 measuring masses between 100-1700 Da with 1 / kO values between 0.6 and 1.6vs / cni2. MS2 scans between 100 and 1700 Da at a resolution of 30,000 was performed on precursor between charge states of 2-5 with targeted intensities of 20000, an intensity threshold of 2500 au and 10 PASEF MS / MS scans were per- formed in each cycle with cycle times of 1.2 sec. The peptides were fragmented using CID with an isolation window of 2 Da and collision energies ranging from 20 eV (1 / kO value of 0.6 Vs / cm2) to 59 eV (1 / kO value of 1.6 Vs / cm2). Dynamic exclusion was enabled for 30 s.

[0387] The LC-MS / MS data were searched against the human SwissProtKB database downloaded on 3-23-2022 (26,576 entries) using the program PEAKSOnline vl 1. These searches were performed considering full LysC peptides with no more than 2 missed cleavage sites. The MSI and MS2 mass accuracies were set to 20ppm and 0.05 Da, respectively, carbamidomethylating was considered as a fixed modification, and oxidation of methionine and protein acetylation were considered as variable modifications. Deep learning boost was enabled. The results were filtered using a reverse decoy database strategy using percolator and the PSM, Peptide, and protein FDR rates were set to 1%. Positively identified proteins were required to be identified by a minimum of 2 peptides with at least one of these being a unique peptide identification.

[0388] Label-free quantitation was performed with PeaksOnline using ID-directed LFQ. Match between runs was enabled with a mass tolerance of 20 ppm, retention time shift set to auto, and a 1 / kO tolerance of 0.05. Quantitation was performed on unique peptides with a feature intensity of at least 150. The LFQ output produced after this step was used for bar graphs in Fig. 4, 8, and the heatmap in Fig. 4F. Data used for Volcano plots in Fig. 4 and8 were further processed as described below. The LFQ output was uploaded into Perseus V2.0.11 and the intensity values were log2 transformed and the data was filtered to remove proteins with <2 quantitative values in at least one of the groups. After matrix reduction, missing values were imputed using a normal distribution with parameters of width = 0.3, downshift = 1.8, and mode = separately for each column. The LFQ intensities were used to determine the abundance ratios of the protein across groups and significance was determined using a two-sided t-test derived p-value (no con’ection for multiple comparisons was performed). Volcano plots in Fig. 4. 8 show proteins that were detected in all three DMSO technical replicates pre-imputation step. Each proteomics experiment consisted of n =3 independent biological replicates per drug condition.Recombinant protein production

[0389] A cDNA sequence corresponding to human TRIM21 PRY-SPRY amino acids 287-465 was cloned into a pET-6X-His vector to generate human 6X-His TRIM21 PRY-SPRY. E. coli BL21 DE3 cells were transformed and induced with 0.4 mM IPTG in TB media supplemented with Ampicillin at OD600 = 0.8 overnight at 18°C. Cleared cell lysates were prepared by sonication of cell pellets in Lysis Buffer (50 mM Tris, 1 M NaCl, 2 mM TCEP, pH=8.0) supplemented with complete protease inhibitors (Roche) followed by centrifugation at 15,000g at 4°C for 1 h. The supernatant was loaded onto a Ni-NTA column and washed with Wash Buffer (50 mM Tris-HCl, 300 mM NaCl, 1 mM TCEP, 20 mM imidazole, pH 8.0). Proteins were eluted with Elution Buffer (50 mM Tris-HCl, 300 mM NaCl, 1 mM TCEP, 400 mM imidazole, pH 8.0). The eluted 6X-His TRIM21 PRYSPRY was further purified by size-exclusion chromatography and fractionated in PBS, pH 7.4. Fractions containing purified protein were pooled and concentrated to -166 pg / mL before being supplemented with 0.5 mM TCEP.IgG pulldown of TRIM21

[0390] For pulldown of endogenous TRIM21 from cell lysates, wild-type (WT) OCL AML-3 cells were washed 2X with PBS before being resuspended in Immunoprecipitation (IP) Buffer composed of 50 mM Tris pH 8.0, 200 mM NaCl, and 0.1% TERGITOL solution (NP40S, Sigma) supplemented with IX cOmplete™ Mini EDTA-free Protease Inhibitor Cocktail (Roche, 11836170001) at a concentration of 3 grams cell pellet weight / mL. Cellsuspensions were lysed by 2X freeze thaw cycles in liquid nitrogen. Lysates were clarified by centrifugation at 20,000g for 20 min at 4°C and soluble protein concentrations were quantified by Pierce BCA Assay (Thermo Fisher Scientific, 23225). Lysates were diluted to 1 mg / niL and 0.5 mg of total soluble protein was loaded onto 20 pl of Anti-FLAG M2 magnetic bead slurry (M8823, Millipore Sigma) with DMSO or indicated compounds. For pulldown of recombinant TRIM21, 20 pg of purified 6X-His TRIM21 PRY-SPRY was diluted in PBS supplemented with 0.5mM TCEP to a concentration of 20 pg / mL and loaded onto 40pl of Anti-FLAG M2 magnetic bead slurry (M8823, Millipore Sigma) with DMSO or indicated compounds. Bead-lysate mixtures were incubated with end-over-end rotation at 4°C for 3 h before washing 5X with IP buffer containing DMSO or indicated compounds. Bead elution was performed by incubation at 95°C in 15 pl of 2X LDS sample buffer (NP0007, Invitrogcn) for 3 min. Eluted proteins were then separated on a 4—12% gradient gel (Invitrogen, NW04125BOX) before being transferred to PVDF membranes using the iBlot 2 Gel Transfer Device (Invitrogen, 1B21001) or stained with InstantBlue Coomassie Protein Stain (Abeam, ISB1L). Membranes were processed as described in the Western Blotting section.Immunofluorescence

[0391] C33a eGFP and TRIM21-FLAG cells were plated at a density of 6000 cells per well in a black, clear-bottom 96-well plate (PerkinElmer, 50-209-9831) and allowed to attach overnight at 37°C. Cells were then treated with respective inhibitors for 6 h. Cells were fixed in 4% PFA for 20 min, washed with PBS, and stained with anti-RANBPl (Abeam, ab97659) at a 1:500 dilution and DAPI (Sigma-Aldrich, D8417) at a 1:20,000 dilution. Cells were imaged using an Operetta High Content Imaging and Analysis System (Perkin Elmer), with eight fields captured per well and two wells per condition at x20 magnification. Images were analyzed using Harmony software on the Columbus data server (PerkinElmer). In brief, live cells were identified and their nuclear regions established using DAPI staining. Around these nuclear regions, the cytoplasmic region was defined according to the region of RANBP1 staining not overlapping with the nucleus. The nuclear-to-cytoplasmic ratio of total signal intensity per well for RANBP1 was calculated based on these criteria.Live fluorescence microscopy (high-content imaging)

[0392] A549 PML-eGFP-BRD4(BD2) TRIM21-FLAG cells were plated at a density of 2000 cells in phenol red-free complete medium in a black, clear-bottom 384-well plate (Falcon, 353962) and allowed to attach over-night at 37°C. Cells were pre -treated with indicated rescue compounds for 2 h and then treated with indicated experimental compounds for 8 h. Cells were stained with Hoechst 33342 (Thermo Fisher Scientific, H3570) 15 min before imaging. Cells were then imaged using an Operetta High Content Imaging and Analysis System (Perkin Elmer), with five fields captured per well and two wells per condition at x20 magnification. Images were analyzed using Harmony software on the Columbus data server (PerkinElmer). In brief, live cells were identified and their nuclear regions established using Hoechst staining. Cell boundaries were established using the eGFP channel and eGFP+ foci were called and quantified using the Harmony Software. Data for each molecule was normalized to DMSO wells.Generating PRLX-93936-resistant cells with CRISPR-Cas9 targeting of NUP98-96

[0393] Short guide RNA oligonucleotides targeting exon 17 of NUP98-96 were synthesized by Synthego. Recombinant SpCas9-2NLS purchased from Synthego was mixed with each guide for 15 min at room temperature to allow for ribonucleoprotein complex formation (20 pmol Cas9:60 pmol single guide RNA per 1 million cells). Each mixture was added to 1 million A549 cells and delivered via electroporation using the Lonza 4D-Nucleofector system and an SF Cell Line 4D-Nucleofector kit according to the manufacturer’s instructions. After electroporation, the cells were allowed to proliferate for 72 h before selection for resistant clones using 1 pM PRLX-93936. After 7 days of selection, the surviving cells were maintained in 1 pM PRLX-93936 and harvested for analysis by western blotting and cell viability assays.Results

[0394] We previously used pharmacological inhibition of neddylation to identify small molecules whose mechanism of cytotoxicity requires Cullin RING Ligase activity. A limitation of this approach is that only the Cullin RING subfamily of E3 ligases are inactivated. We imagined that inactivating a wider range of E3 ligases could potentially spotlight additional degraders whose mechanisms target non-CRL E3 ligases. Small-molecule inhibition of the ubiquitin-like modifier activating enzyme (UBA1 / UAE) prevents activation of the large majority of E3 ligases and is currently being pursued as an anticancer strategy. While sustained treatment with the UBA1 inhibitor TAK-243 was potently cytotoxic to the acute myeloid leukemia cell line OCI- AML-3 (EC50 15 nM), treatment for 10 h was better-tolerated (Fig. 7A). Short-term treatment with TAK-243 thus provided a meaningful window during which cells could be maintained in a viable state without E3 ligase activity.

[0395] We next established a comparative cell viability assay in 384-well plates in which the impact of small molecules was compared with and without the addition of TAK-243; we then evaluated 1754 known bioactive small molecules using this assay (Figs. 1 A, B;). Only 5 molecules were identified that showed <50% viability and gained at least 30% viability with TAK-243 co-trcatmcnt (Fig. IB). Hit validation across a wide concentrationresponse range confirmed only one of the five, PRLX-93936, as having diminished cytotoxicity when co-treated with TAK-243 (Fig. 1C, D; Fig. 7B). This low confirmed hit rate likely reflects both the uncommon nature of ubiquitination-dependent cytotoxic mechanisms and the requirement for induction of cell death within 10 h. PRLX-93936 cytotoxicity could also be rescued by the proteasome inhibitor bortezomib, supporting a mechanism of cytotoxicity dependent on the proteasome (Fig. ID).

[0396] PRLX-93936 is a cytotoxin of unknown mechanism of action that has been linked to diminished HIF pathway signaling. Publicly-available cancer cell line profiling data (depmap.org) indicated that PRLX-93936 induced wide-ranging cytotoxic responses in cancer cell lines, with many potently killed and others fully resistant (Fig. 7C). Correlation of cell killing potency with expression of single transcripts has previously provided mechanistic understanding for molecules of unknown mechanism-of-action, including the molecular glue CR8. Analysis of publicly available transcriptomic profiles revealed that sensitivity to PRLX-93936 was uniquely correlated with the expression of TRIM21, a member of the TRIM family of E3 ligases (Fig. IE, Fig. 7C). Conversely, among the thousands of small molecules in the DepMap database, PRLX-93936 was best-correlated with TRIM21 transcript levels, highlighting the strong association between PRLX-93936 and TRIM21 (Fig. IF).

[0397] A second molecule, the famesyl transferase inhibitor (FTI) BMS-214662, also appeared well-correlated with TRIM21 expression level (Fig. IF, G). Interestingly, workfrom Bristol Myers Squibb and others has established that this molecule has an additional uncharacterized apoptotic mechanism not observed for closely related FTI inhibitors, including its X-methyl analog BMS-225975 (Fig. 1G). We next confirmed that BMS-214662, like PRLX-93936, induced rapid cell death in OCI-AML-3 cells that could be suppressed by co-treatment with either TAK-243 or bortezomib (Fig. 1H). Together these data suggested that PRLX-93936 and BMS-214662 share a cytotoxic mechanism of action that involves TRIM21 -mediated ubiquitination.

[0398] We next confirmed strong expression of TRIM21 in both JURKAT and OCI-AML-3 cells and used CRISPR / Cas9 targeting to knockout TRIM21 in both cell lines (Fig. 2A, B). Strikingly, while WT cells were highly sensitive to PRLX-93936 (ECso ca. 100 nM), TRIM21 KO in both Jurkat and OCI-AML-3 cells led to lull resistance to PRLX-93936 at concentrations as high as 50 pM (Fig. 2C, D; Fig. 8A). TRIM21 KO had no discernible impact on proliferation in these cell lines, consistent with TRIM21 not being a known dependency across hundreds of cancer cell lines (Fig. 8B, depmap.org). Likewise, both TRIM21 KO cell lines showed >100-fold increase in ECso for BMS-214662 (Fig. 2E, F); farnesyl transferase inhibition may explain cell killing observed at high concentrations. Notably, while PRLX-93936 shares some structural features with erastin, an inducer of ferroptosis, erastin sensitivity was unaffected by TRIM21 KO (Fig. 2G). The cytotoxicity of farnesyl transferase inhibitor BMS-225975, which differs from BMS-214662 by only addition of a methyl group, was also unaffected by TRIM21 KO (Fig. 2H).

[0399] We also evaluated the impact of overexpression of TRIM21 on PRLX-93936 and BMS-214662 sensitivity. OCI-AML-3 cells expressing a TRIM21-FLAG allele became ca. 10-fold more sensitive to both PRLX-93936 and BMS-214662 (Fig. 3A-C). TRIM21 expression had no discernible impact on proliferation (Fig. 8C), also consistent with prior reports that stable TRIM21 overexpression is non-toxic. In contrast, expression of an established catalytically inactive triple TRIM21 mutant (C16A / C31A / H33W; TRIM21CA) partially suppressed the cell killing induced by these probes (Fig. 3A-C). Since TRIM21 enzymatic activity is known to be enhanced by its oligomerization, this catalytically inactive allele may induce a dominant-negative effect and partially mimic TRIM21 loss-of-function. We also overexpressed TRIM21 in C33A cells, a cervical cancer line that both publicly available transcriptomics data (www.depmap.org) and immunoblotting support as not expressing TRIM21 (Fig. 3D). While parental C33A cells were resistant to PRLX-93936and BMS-214662 at concentrations up to 10 pM. overexpression of TRIM21 greatly sensitized C33A cells to these probes, with EC50 values comparable to those seen in WT OCI-AML-3 or Jurkat cells (ca. 100 nM; Fig. 3E, F). As expected, expression of the catalytically inactive TRIM21 CA mutant allele had no impact on sensitivity (Fig. 3E, F). As a final example, overexpression of TRIM21 in HEK293T cells also induced extraordinary sensitivity to PRLX-93936 and BMS-214662 (Fig. 3G-I). Together these gain- and loss-of-function genetic manipulations indicate that TRIM21 catalytic activity is both necessary and sufficient for PRLX-93936 and BMS-214662 to induce cell death.

[0400] To evaluate whether these small molecules directly bind TRIM21, we performed cellular thermal shift assays (CETSA). PRLX-93936 induced clear stabilization of TRIM21-FLAG by 1-2°C, consistent with a direct binding interaction (Fig. 3J; Fig. 8D). BMS-214662 stabilized TRIM21-FLAG by a larger margin, providing additional evidence that these molecules induce cell death by the direct binding to TRIM21 and subsequent ubiquitination and proteasomal degradation of an essential protein (Fig. 3K, Fig. 8E). As recently-reported TRIM21 -targeting glues have been demonstrated to bind to TRIM21’s PRYSPRY domain, we hypothesized that PRLX-93936 and BMS-214662 also likely bound within this established pocket. To support this idea, we immobilized endogenous TRIM21 from cell lysates onto IgG-coated resin; IgG is a high-affinity endogenous ligand of the TRIM21 PRYSPRY domain. Treatment with 1, a close analog of PRLX-93936 (Fig. 11), or BMS-214662 dose-responsively displaced TRIM21 from the resin, supporting a competitive interaction with the TRIM21 PRYSPRY domain (Fig. 3L; Fig. 8F, G). Additionally, we observed that HGClg, a recently reported high-affinity ligand for the TRIM21 PRYSPRY domain that lacks molecular glue properties, fully and dose-responsively suppressed the cytotoxicity of PRLX-93936 and 1 (Fig. 8H). Together these studies establish PRLX-93936 as directly engaging TRIM21 at its PRYSPRY domain.

[0401] To identify the protein or proteins degraded by PRLX-93936 and BMS-214662, we next performed an unbiased proteomic analysis. We initially evaluated PRLX-93936 treatment in both JURKAT and OCI-AML-3 cells to identify proteins comparably depleted across these equally sensitive cell lines. Strikingly, a cluster of nucleoporin proteins were strongly downregulated in both JURKAT and OCI-AML-3 cells (Fig. 4A, B, Fig. 9A, B). Strongest effects were seen for Nucleoporin 214 (NUP214) and Nucleoporin 88 (NUP88), which are essential proteins known to directly interact at the cytoplasmic face of the nuclearpore (Fig. 4B, D). However, numerous additional essential nucleoporin proteins-NUP98, NUP188, NUP155, and NUP35, among others-were also reduced (Fig. 4A, Fig. 9C).Importantly, changes in NUP214 and NUP88 levels were not observed when the proteomics experiment was repeated using TRIM21 KO OCI- AML-3 cells, indicating that the reduction of these nucleoporins is TRIM21 -dependent (Fig. 4C, E). Additionally, proteomic analysis revealed that nucleoporin degradation was rapid, with levels of some nucleoporins reduced by half within an hour (Fig. 4F, Fig. 9E).

[0402] We then repeated these proteomics experiments with BMS-214662. As with PRLX-93936, multiple nucleoporins were degraded by BMS-214662 after 4 h in OCI-AML-3 cells, with NUP88 and NUP98 most strongly affected (Fig. 4G, H). Critically, nucleoporin levels were unaffected by BMS-214662 treatment in OCI-AML-3 cells lacking TRIM21, further supporting TRIM21’s essential role in mediating nucleoporin degradation (Fig. 41, Fig. 9F). Moreover, BMS-225975, which differs from BMS-214662 only by addition of a methyl group but lacks TRIM21 -dependent cell killing (Fig. 2H), had no impact on nucleoporin levels (Fig. 9G).

[0403] We also used western blotting as an orthogonal approach to validate our proteomics results. Levels of both NUP214 and NUP88 were substantially depleted following treatment with PRLX-93936 for 4 h in OCI- AML-3 cells (Fig. 4J). In contrast, reductions in NUP214 and NUP88 were not observed when PRLX-93936 was co-treated with the El inhibitor TAK-243 or the proteasome inhibitor bortezomib, indicating the nucleoporin loss was ubiquitin- and proteasome-dependent (Fig. 4K).

[0404] Loss of numerous essential nucleoporin proteins would be expected to impair nuclear trafficking. We next evaluated nuclear export in C33A cells using immunofluorescence detection of RANBP1 as a common marker of nuclear- to-cytoplasmic trafficking. C33A cells express TRIM21 at undetectable levels and are resistant to PRLX-93936 / BMS-214662 unless TRIM21 is overexpressed (Fig. 3D-F). In C33A cells, PRLX-93936 and BMS-214662 had no impact on the sub-cellular localization of RANBP1; however, a canonical nuclear export inhibitor, the XPO1 -targeting natural product Leptomycin B, led to strong nuclear accumulation of RANBP1 (Fig. 4L, M, Fig. 9H). In contrast, treatment of TRIM21-overexpressing C33A cells with multiple concentrations of either PRLX-93936 or BMS-214662 induced nuclear accumulation of RANBP1 comparable to Leptomycin B treatment (Fig. 4L, M, Fig. 9H). Additionally, nuclear accumulation ofRANBP1 was not observed in cells overexpressing catalytically inactive TRIM21 (Fig. 4L). Importantly, this TRIM21 -dependent nucleoporin degradation mechanism helps to rationalize past unexplained observations of extensively altered subcellular protein localization following BMS-214662 treatment. Together, these data highlight that the observed loss of multiple essential nucleoporins leads to lethal nuclear trafficking deficits that are dependent on catalytically active TRIM21.

[0405] Recent studies have also noted TRIM21 -mediated degradation of nucleoporins by small molecules including hydroxy-acepromazine and HGC652. These studies also provided genetic evidence that hydroxy-acepromazine induced degradation of multiple nucleoporins by recruiting TRIM21 specifically to NUP98. We confirmed that hydroxy-acepromazine indeed caused TRIM21 -dependent cell death in OCI- AML-3 cells, albeit with 100-fold diminished potency relative to PRLX-93936 and BMS-214662 (Fig. 5B, C).Additionally, we established that targeted disruption of the autoproteolysis domain of NUP98 was sufficient to prevent cell killing by PRLX-93936 and BMS-214662. Delivery of recombinant Cas9 and five independent single-guide RNAs (sgRNAs) targeting this region of NUP98 led to isolation of A549 cells that were strongly resistant to both PRLX-93936 and BMS-214662 (Fig. 5D-E). These resistant cells did not show resistance to the unrelated cytotoxic agent TAK-243, supporting a selective effect (Fig. 5F). Each of these five distinct sgRNAs led to detection of a 200 kDa NUP98 proteoform in these PRLX-93936-resistant cells, indicating that the autoproteolysis of NUP98 had been impaired (Fig. 5G). Impaired autoproteolysis of NUP98 was recently demonstrated as a resistance mechanism to hydroxy-acepromazine. Finally, these PRLX-93936-resistant cells did not show reduction in NUP214 or NUP88 levels following treatment with PRLX-93936, providing additional support for nucleoporin degradation as essential for this cytotoxic mechanism (Fig. 5H).

[0406] We then performed medicinal chemistry optimization on the PRLX-93936 template both to identify analogs with improved potency and to elaborate PRLX-93936 to TRIMTACs that degrade a cellular protein aggregate. We synthesized a collection of 30 PRLX-93936 derivatives and evaluated their ability to induce TRIM21 -mediated cell death in both OCI- AML-3 and JURKAT cells (see Fig. 11 for data for all analogs). Initial analogs made modifications to the piperazine ring of PRLX-93936. Paralleling results observed during evaluation of PRLX-93936 analogs as HIF1 pathway antagonists, we found that modifications that altered the terminal piperazine amine functionality, either by alkylation,substitution with oxygen, or conversion to an amide, strongly abrogated potency (Fig. 6 A, Fig. 11). However, analogs that added a methyl substituent to the piperazine carbon backbone were tolerated, with some leading to clear improvement in cell killing potency (Fig. 6A, Fig. 11). Separation of methyl diastereomers 1 and 8 revealed that the (S)-methyl-piperazine 1 showed clearly improved potency relative to the (7 )-configured 8 and represents the most potent TRIM21 -targeting glue in this series (EC503 nM for cytotoxicity to OCI-AML-3) (Fig. 6A). Additionally, each cytotoxic analog was evaluated in TRIM21 KO cells, which were uniformly resistant up to 2 pM, supporting TRIM21’s essential role in the observed cell killing (Fig. 11).

[0407] Additionally, in confirming the enantiopurity of these analogs by analytical chiral SFC, we noted that these molecules in fact were a racemic mixture of stereoisomers. Past reports for related quinazolinone systems and calculations performed on PRLX-93936 have established that these molecules exist as atropisomers with rotational barriers of >35 kcal / mol, making them indefinitely stable near room temperature. We used preparatory chiral SFC to separate the two PRLX-93936 atropisomers (9 and 10) and confirmed that no inter-conversion occurred over 96 h (Fig. 10A). Strikingly, all cellular activity resided in one atropisomer, which was roughly twice as potent as the racemic mixture of PRLX-93936 (Fig. 6B). Similarly, only one atropisomer showed cell killing among the enantiomeric pair of 11 and 12 (Fig. 10C). Remaining analogs were evaluated as racemates unless noted.

[0408] Further analysis of modified piperazine-containing analogs revealed that substitution of the piperazine methyl group of 1 with ethyl or isopropyl was tolerated but somewhat less potent than 1 (Fig. 11). In contrast, numerous analogs that added a second methyl group around the piperazine were inactive (Fig. 11). Additionally, while either (R)-or (S)-methyl substitution of the piperazine was tolerated, gem-dialkyl substitution at this position was deleterious, as cyclopropyl analog 18 showed greatly diminished potency relative to 1.

[0409] We also evaluated modifications to the quinazolinone and aryl rings of PRLX-93936. Substitution of the quinazolinone with methyl at each of the available four positions led to analogs with diminished potency, with substitution at the 5- and 6-positions somewhat less disruptive than substitution at the 7- and 8-positions (Fig. 10B). In contrast, when methyl was added to each of the four positions of the aryl ring, a wide spectrum of outcomes was observed. Methyl substitution at the 3-position was not tolerated; addition at the 5- or 6-positions led to modest losses in potency; and addition of methyl at the 4-position led to a 3-fold gain in potency relative to PRLX-93936 (Fig. 6C, Fig. 11). Subsequent combination of the (S')-methyl piperazine (as in 1) with the 3-methyl moiety of 24 gave 27, which did not show further potency increase relative to 1 alone (Fig. 11). Notably, bulkier isopropyl substituents could be accommodated at C4, C5 and C6 of the aryl ring, highlighting that this region is broadly tolerant of expansion (Fig. 11).

[0410] We next sought to evaluate whether the PRLX-93936 scaffold could be leveraged for the development of PROTACs that degrade protein aggregates. While our SAR data highlighted multiple positions where substituents could be added while improving potency, it remained unclear which positions could successfully accommodate the necessary linker moiety and facilitate ternary complex formation. We generated an initial set of five heterobifunctionals that connected PRLX-93936 via a flexible linker to JQ-1, a ligand used frequently in PROTACs to promote degradation of BET bromodomain proteins. Of these five, three added the linker moiety to piperazine functionality, while one added the linker at C5 of the aryl ring, which uniquely led to separable diastereomeric atropisomers 34 and 35 that were evaluated separately (Fig. 10D). To test these candidate PROTACs, we used a recently-reported assay in which bromo-domain 2 of the BRD4 protein is fused both to PML, a protein known to form nuclear condensates, and enhanced green fluorescent protein (EGFP). Expression of this fusion protein in A549 cells also expressing wild-type TRIM21 gave rise to cells with prominent green nuclear puncta, indicative of the expected aggregation of the fusion protein (Fig. 6D). Treatment of these cells for 8 h with 31, 32, or 33 had little to no impact of EGFP+ nuclear puncta; in contrast, 34 gave near-complete loss of EGFP+ nuclear puncta with an EC50 of 1.4 pM (Fig. 6D, E, Fig. 10E, F). Notably, no such degradation was observed in cells treated with diastereomeric atropisomer 35, highlighting the stereospecificity of this effect (Fig. 6D, E, Fig. 10E, F). Additionally, the TRIM21 ligand HGClg or the proteasome inhibitor bortezomib were sufficient to prevent degradation of the EGFP fusion protein, confirming TRIM21 -mediated proteasomal degradation of this aggregation-prone fusion protein by the PRLX-93936-derived TRIM-TAC 34 (Fig. 6F, G, Fig. 10G).

[0411] We have established that PRLX-93936 and BMS-214662 induce cell death via TRIM21 -mediated degradation of nucleoporins. Genetic manipulations support TRIM21 as necessary and sufficient for the observed cell death across multiple cell lines. Extensiveproteomic analyses identified a wide range of nucleoporins as degraded following treatment with PRLX-93936 and BMS-214662. Additionally, CRISPR targeting of NUP98’s autoproteolysis domain provided strong resistance to these small molecules. This finding supports that NUP98 plays an essential role in the observed nucleoporin degradation and may directly contact TRIM21 in the presence of PRLX-93936 and BMS-214662. Notably, NUP98 levels were unchanged in TRIM21 KO cells (Fig. 9D), supporting that TRIM21 does not regulate NUP98 levels in the absence of small molecule glues.

[0412] Very recent work has established other small molecules that induce cell death via TRIM21 -mediated nucleoporin degradation, including hydroxy-acepromazine and HGC652. These molecules, like PRLX-93936 and BMS-214662, share the ability to degrade multiple nucleoporins and other proteins, demonstrating how loss of nuclear pore function can induce broad and rapidly lethal disruption of the proteome. Interestingly, these four series of TRIM21 -targeting glues share almost no structural similarity, highlighting how distinct scaffolds can equally function to induce nucleoporin degradation. Notably, analog 1 offers substantially enhanced potency for cell killing relative to hydroxy-acepromazine or HGC652.

[0413] A unique aspect of PRLX-93936 and BMS-214662 is that these molecules have previously been investigated clinically in cancer. While clinical data for PRLX-93936 have only been presented at conferences, published reports indicate that BMS-214662 was well tolerated and showed robust objective responses in a subset of acute myeloid leukemia and myelodysplastic syndrome patients. Apoptotic responses were observed in tumors following treatment with BMS-214662 but not with other famesyl transferase inhibitors, suggesting that the observed clinical responses were unlikely to result from farnesyl transferase inhibition. Together with our observation that high expression of TRIM21 strongly predicts PRLX-93936 and BMS-214662 sensitivity (Fig. 1), these findings suggest the potential for clinical re-evaluation of these agents in subsets of patients whose tumors have elevated TRIM21 expression.

[0414] Recent work with hydroxy-acepromazine has demonstrated that TRIM21 targeting glues can be elaborated to TRIMTACs that degrade proteins engineered to reside within nuclear condensates. However, the low affinity of hydroxy-acepromazine for wildtype TRIM21 necessitated the use of a mutant TRIM21 allele to observe protein degradation. Our optimization of highly potent analogs of PRLX-93936 has now enabled us to identify aTRIMTAC capable of leveraging wild-type TRIM21 to degrade an aggregated protein.TRIM21’s unique biological preference for ubiquitination of antibody-coated pathogens and other large multimeric assemblies suggests that TRIM21 may be ideally suited for degrading aggregated proteins. Establishing PRLX-93936 and BMS-214662 as highly potent TRIM21-targeting molecular glues and evolving PRLX-93936 into a TRIMTAC (34, Fig. 6H) creates opportunities for modulating TRIM21 for cancer therapy and in a wide range of targeted protein degradation applications.Example 2Chemical Characterization data for synthesized small moleculesCW-1

[0415] To a solution of the indicated aniline (200 mg, 1.46 mmol, 190.84 pL) and indicated chloroacetamide (311.44 mg, 1.46 mmol) in ACN (3 mL) was added POCL (670.65 mg, 4.37 mmol, 407.69 pL). The mixture was stirred at 80°C for 12 hrs. LC-MS showed the aniline was consumed and 37% of desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by flash silica gel chromatography (ISCO®; 10 g SepaFlash® Silica Flash Column, Eluent of 15% Ethyl acetate / Petroleum ether gradient @ 60 mL / min) to afford Compound 1 (400 mg, 1.27 mmol, 87.16% yield) as a red oil. MS-ESI (m / z) calcd for C17H15CIN2O2 [M+H]+: 315.1 / 317.1 Found 315.1 / 317.1.

[0416] To a solution of Compound 1 (100 mg, 317.70 pmol) and Compound lb (76.35 mg, 381.24 pmol) in DMF (2 mL) was added K2CO3 (131.72 mg, 953.09 pmol). The mixture was stirred at 80°C for 12 hrs. LC-MS showed Compound 1 was consumed and 43% of desired compound was detected. The reaction mixture was diluted with H2O 10 mL and extracted with EtOAc (10 mL * 3). The combined organic layers were washed with brine (10 mL * 2), dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressureto give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 4 g SepaFlash® Silica Flash Column, Eluent of 20% Ethyl acetate / Petroleum ether gradient @ 50 mL / min) (SiO2, Petroleum ether: Ethyl acetate = 5:1, Rf (Pl) = 0.40) to afford Compound 2 (80 mg, 167.16 pmol, 52.62% yield) as a pale yellow oil. MS-ESI (m / z) calcd for C27H34N4O4 [M+H]+:479.2 / 480.2 Found 479.3 / 480.3.

[0417] To a solution of Compound 2 (80 mg, 167.16 pmol) in DCM (1 ml.) was added TFA (307.00 mg, 2.69 mmol, 0.2 mL). The mixture was stirred at 20°C for 1 hr. LC-MS showed Compound 2 was consumed and 71 % of desired compound was detected. The reaction mixture was concentrated under vacuum. The residue was purified by Prep-HPLC (column: Phenomenex Luna C18 100*30mm*5um; mobile phase: [H2O (0.1% TFA) - ACN]; gradient: 5%-40% B over 8.0 min) to afford CW-1 (67.99 mg, 138.05 pmol, 82.59% yield, 100% purity, TFA salt) as a colorless gum.Spectrum:

[0418] ¹H NMR (METHANOL-d4, 400 MHz): 5 ppm 8.19 - 8.28 (m, 1 H) 7.84 - 7.97 (m, 1 H) 7.78 (dd, 7=8.13, 2.00 Hz, 1 H) 7.56 - 7.63 (m, 1 H 7.48 - 7.56 (m, 1 H) 7.37 (td, 7=7.41, 1.56 Hz, 1 H) 7.23 (br d, 7=8.50 Hz, 1 H) 7.14 (td, 7=7.63, 1.25 Hz, 1 H) 4.03 - 4.22 (m. 2 H) 3.75 - 3.96 (m, 1 H) 3.37 -3.28 (m, 1 H) 3.15 - 3.24 (m, 1 H) 3.06 - 3.15 (m, 1 H) 2.74 - 3.04 (m, 3 H) 2.64 - 2.74 (m, 1 H) 2.51 - 2.64 (m, 1 H) 1.22 (td, 7=7.00, 2.88 Hz, 3 H) 0.75 (dd, 7=6.32, 1.69 Hz, 3 H).

[0419] ¹³C NMR (METHANOL-d4, 101 MHz): 5 ppm 162.3, 162.0, 154.9, 154.2, 154.0, 153.9, 146.6, 134.8, 129.3, 127.3, 127.2, 126.7, 126.6, 126.4, 125.4, 120.7, 120.5, 113.0, 112.9, 64.2, 64.0, 52.5, 48.5, 43.2, 43.1, 13.9, 13.8, 13.6, 13.5

[0420] LCMS (ESI+): m / z 379.2 (M+H)

[0421] HRMS (TOF MS ES+): calcd for C22H27N4O2 [M+H]+: 379.2134 Found 379.2119.CW-2K2CO3, DMFCW-2

[0422] To a solution of Compound 1 (100 mg, 317.70 pmol) and N-methylpiperazine (1A) (31.82 mg, 317.70 pmol, 35.24 pL) in DMF (2 m ) was added K2CO3(131.72 mg, 953.09 pmol). The mixture was stirred at 80°C for 2 hrs. LC-MS showed Compound 1 was consumed and 32% of desired compound was detected. The reaction mixture was filtered. The filtrate was purified by Prep-HPLC (column: Phenomenex Luna C 18 100*30mm*3 pm; mobile phase: [H2O (0.1 % TFA) - ACN]; gradient: 5% - 40% B over 8.0 min) to afford CW-2 (50 mg, 101.52 pmol, 31.96% yield, TFA salt) as a white solid.

[0423] NMR (METHANOL- 4, 400 MHz): 5 ppm 8.24 (dd, J = 1.1, 8.0 Hz, 1H), 7.93 - 7.86 (m, 1H), 7.78 (d, J = 8.1 Hz, 1H), 7.60 (t,.1 = 7.6 Hz, 1H), 7.56 - 7.49 (m, 1H), 7.42 (dd, J = 1.5, 7.8 Hz, 1H), 7.24 (d, J = 7.9 Hz, 1H), 7.15 (t, J = 7.6 Hz, 1H), 4.21 - 4.03 (m, 2H), 3.50 - 3.35 (m, 4H), 3.15 - 2.82 (m, 6H), 2.68 - 2.47 (m, 3H), 1.21 (t, J = 7.0 Hz, 3H). LCMS (ESI+): m 379.2 (M+H).CW3K2CO3, DMF1 CW-3

[0424] To a solution of Compound 1 (90 mg, 285.93 pmol) and Compound 1 A (99.00 mg, 1.14 mmol, 0.1 mL) in DMF (2 mL) was added K2CO3 (118.55 mg, 857.78 pmol). The mixture was stirred at 80°C for 2 hrs. LC-MS showed Compound 1 was consumed and 46% of desired compound was detected. The reaction mixture was filtered. The filtrate was purified by Prep-HPLC (column: Phenomenex Luna C18 100*30mm*3 pm; mobile phase:[H2O (0.1 % TFA) - ACN]; gradient: 5% - 35% B over 8.0 min) to afford CW-3 (110.49 mg, 229.74 pmol, 80.35% yield, 99.69% purity, TFA salt) as a white solid.

[0425] NMR (METHANOL-6Z4, 400 MHz): 5 ppm 8.21 - 8.30 (m, 1 H), 7.83 - 7.99 (m. 2 H). 7.56 - 7.69 (m, 2 H), 7.40 (dd, J=7.75. 1.50 Hz, 1 H), 7.29 (br d,.1=8.50 Hz, 1 H), 7.20 (br t,.1=7.63 Hz, 1 H), 4.32 (br d, J=16.88 Hz, 1 H), 4.16 - 4.24 (m, 1 H), 4.06 - 4.14 (m, 1 H), 4.05 (br s, 5 H), 3.32 - 3.69 (m, 4 H), 1.22 (t, J=6.94 Hz, 3 H)

[0426] LCMS (ESI+): m!z 366.2 (M+H).CW-4

[0427] To a solution of Compound 1 (90 mg, 246.95 pmol; see CW-9 / CW-10 below) in DCM (1 mL) was added TEA (74.97 mg, 740.86 pmol, 103.12 pL) and AcCl (38.77 mg, 493.91 pmol, 35.12 pL) at 20°C. The mixture was stirred at 20°C for 1 hr under N2. LC-MS showed Compound 1 was consumed and 97% of desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by Prep-HPLC (TFA condition; column: Phenomenex Luna C18 100*30mm*3 pm; mobile phase: [H2O (0.1% TEA) - ACN]; gradient: 5% - 35% B over 8.0 min) to afford CW-4 (116.07 mg, 222.02 pmol, 89.90% yield, 99.56% purity, TFA salt) as a white solid.

[0428] 1H NMR (METHANOL-r / 4, 400 MHz): 5 ppm 8.31 - 8.24 (m, 1H), 7.98 - 7.91 (m, 1H), 7.90 - 7.86 (m, 1H), 7.68 - 7.62 (m, 1H), 7.62 - 7.55 (m, 1H), 7.40 (dd, J = 1.4, 7.71C Hz, 1H), 7.29 (d, J = 8.3 Hz, 1H), 7.19 (t, J = 7.7 Hz, 1H), 4.30 - 4.21 (m, 1H), 4.20 - 4.04 (m, 2H), 4.02 - 3.78 (m, 5H), 3.51 - 3.34 (m, 4H), 2.14 (s, 3H), 1.22 (t, J = 6.9 Hz, 3H) LCMS (ESI+): m / z 407.2 (M+H).CW-51C HK2CO3, DMFCW-5

[0429] To a solution of Compound 1 (100 mg, 317.70 pmol) and Compound 1C (34.99 mg, 349.47 pmol) in DMF (2 mL) was added K2CO3 (131.72 mg, 953.09 pmol). The mixture was stirred at 80°C for 12 hrs. LC-MS showed Compound 1 was consumed and 22% of desired compound was detected. The reaction mixture was filtered. The filtrate was purified by Prep-HPLC (column: Phenomenex Luna C18 100*40mm*5 pm; mobile phase:[II2O (0.1% TFA) - ACN]; gradient: 5% - 35% B over 8.0 min) to afford CW-5 (43.6 mg, 86.89 pmol, 27.35% yield, 98.14% purity, TFA salt) as a white solid.

[0430] 1H NMR (METHANOL-t / 4, 400 MHz):5 ppm 8.22 - 8.29 (m, 1 H), 7.88 - 7.98 (m, 1 H), 7.81 - 7.86 (m, 1 H), 7.63 (t, J=7.38 Hz, 1 H), 7.52 - 7.59 (m, 1 H), 7.39 (dd, J=7.75, 1.50 Hz, 1 H), 7.25 (d, J=8.25 Hz, 1 H), 7.16 (t, J=7.57 Hz, 1 H), 4.05 - 4.19 (m, 2 H), 3.93 - 4.00 (m, 1 H), 3.80 - 3.87 (m, 1 H), 3.57 (s, 2 H), 3.45 (t, J=5.44 Hz, 2 H), 3.12 -3.26 (m, 2 H), 1.22 (t, J=7.00 Hz, 3 H)

[0431] LCMS (ESI+): m / z 379.2 (M+H).CW-6TFA, DCM K2CO3, DMF1 2

[0432] To a solution of Compound 1 (100 mg, 317.70 pmol) and Compound 1A (63.63 mg, 317.70 pmol) in DMF (2 mL) was added K2CO3 (131.72 mg, 953.09 pmol). The mixture was stirred at 80°C for 1 hr. LC-MS showed Compound 1 was consumed and 32% of desired compound was detected. The reaction mixture was diluted with H2O 5 mF and extracted with EtOAc (5 mL * 3). The combined organic layers were washed with brine (5 mb * 2), dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to afford Compound 2 (150 mg, crude) as a yellow oil. MS-ESI (m / z) calcd for C27H34N4O4 [M+H]+: 479.2 Found 479.3.

[0433] To a solution of Compound 2 (150 mg, 313.43 pmol) in DCM (2 mL) was added TFA (767.50 mg, 6.73 mmol, 0.5 mL). The mixture was stirred at 20°C for 1.5 hrs. LC-MS showed Compound 2 was consumed and 50% of desired compound was detected. The reaction mixture was concentrated under vacuum. The residue was purified by Prep-HPLC (column: Phenomenex luna Cl 8 100*40mm*5 pm; mobile phase: [H2O (0.1% TFA) -ACN]; gradient: 10% - 40% B over 8.0 min). The LCMS purity was not high enough, then the residue was purified by Prep-HPLC (column: Waters Xbridge BEH Cl 8 100*30mm*10 pm; mobile phase: [H2O (lOmM NH4HCO3) - ACN]; gradient: 10% - 50% B over 8.0 min) to afford CW-6 (29.24 mg, 59.37 pmol, 18.94% yield, 100% purity, TFA salt) as a colorless gum.

[0434] NMR (METHANOL- 4, 400 MHz): 5 ppm 8.22 (d, J=8.00 Hz, 1 H), 7.83 -7.95 (m, 1 H), 7.77 (d, J=8.13 Hz, 1 H), 7.58 (t, J=7.57 Hz, 1 H), 7.49 (t, J=7.88 Hz, 1 H), 7.36 (d, J=7.63 Hz, 1 H), 7.18 (d,.1=8.25 Hz, 1 H), 7.06 - 7.15 (m, 1 H), 4.01 - 4.16 (m, 2 H), 3.32 - 3.39 (m, 1 H), 3.16 - 3.27 (m, 1 H), 2.71 - 2.80 (m, 1 H), 2.57 - 2.71 (m, 2 H), 2.47 (brd, J=11,01 Hz, 1 H), 2.29 - 2.38 (m, 1 H), 1.96 - 2.07 (m, 1 H), 1.69 (td,.1=10.47, 4.19 Hz, 1 H), 1.20 (t, J=6.94 Hz, 3 H), 0.92 (d, J=6.50 Hz, 3 H)

[0435] LCMS (ESI+): m / z 379.2 (M+H).CW-7TFA, DCM

[0436] To a solution of Compound 1 (100.00 mg, 317.70 pmol) and Compound 1A (63.63 mg, 317.70 pmol) in DMF (2 mL) was added K2CO3 (131.72 mg, 953.09 pmol). The mixture was stirred at 80°C for 1 hr. LC-MS showed 4% of Compound 1 was remained and 13% of desired compound was detected. The reaction mixture was diluted with H2O 5 mL and extracted with EtOAc (5 mL * 3). The combined organic layers were washed with brine (5 mL * 2), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 10 g SepaFlash® Silica Flash Column, Eluent of 20% Ethyl acetate / Petroleum ether gradient @ 50 mL / min) to afford Compound 2 (80 mg, 167.16 pmol, 52.62% yield) as a yellow oil. MS-ESI (m / z) ealed for C27H34N4O4 [M+H]+: 479.2 Found 479.4.

[0437] To a solution of Compound 2 (80 mg, 167.16 pmol) in DCM (1 mL) was added TFA (307.00 mg, 2.69 mmol, 0.2 mL). The mixture was stirred at 20°C for 1 hr. LC-MS showed Compound 2 was consumed and 35% of desired compound was detected. The reaction mixture was concentrated under vacuum. The residue was purified by Prep-HPLC (column: WePure Biotech XP tC18 150*40*7um; mobile phase: [H2O (lOmM NH4HCO3) -ACN]; gradient: 10% - 40% B over 8.0 min) to afford CW-7 (4.98 mg, 9.81 pmol, 5.87% yield, 97.0% purity, TFA salt) as a white solid.

[0438] ¹H NMR (METHANOL-d4, 400 MHz):5 ppm 8.22 (br d, J=7.88 Hz, 1 H), 7.83 - 7.91 (m, 1 H), 7.77 (br d, J=8.13 Hz, 1 H), 7.57 (brt, J=7.44 Hz, 1 H), 7.49 (br t, J=7.82 Hz, 1 H), 7.27 - 7.38 (m, 1 H), 7.15 - 7.23 (m, 1 H), 7.06 - 7.15 (m, 1 H), 4.02 - 4.19 (m, 2H), 3.67 - 3.95 (m, 1 H), 3.02 - 3.22 (m, 1 H), 2.57 - 2.89 (m, 4 H), 2.12 - 2.42 (m, 3 H), 1.13 - 1.27 (m, 3 H), 0.54 - 0.65 (m, 3 H)

[0439] LCMS (ESI+): m / z 379.3 (M+H).CW-8BocH TFA, DCM K2CO3, DMFCW-8

[0440] To a solution of Compound 1 (100 mg, 317.70 pmol) and Compound 1A (95.44 mg, 476.55 pmol) in DMF (2 mL) was added K2CO3 (131.72 mg, 953.09 pmol). The mixture was stirred at 80°C for 12 hrs. LC-MS showed Compound 1 was consumed and 34% of desired compound was detected. The reaction mixture was diluted with H2O 3 mL and extracted with EtOAc (3 mL * 3). The combined organic layers were washed with brine (3 mL * 1), dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to afford Compound 2 (150 mg, crude) as a yellow oil. MS-ESI (m / z) calcd for C27H34N4O4 [M+H]+: 479.3 Found 479.3.

[0441] To a solution of Compound 2 (150 mg, 313.43 pmol) in DCM (2 mL) was added TFA (714.76 mg, 6.27 mmol, 465.64 pL). The mixture was stirred at 20°C for 1 hr. LC-MS showed Compound 2 was consumed and 74% of desired compound was detected.The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by Prep-HPLC (TFA condition; column: Phenomenex Luna Cl 8 100*30mm*5 pm; mobile phase: [H2O (0.1% TFA) - ACN]; gradient: 5% - 40% B over 8.0 min) to afford CW-8 (80.56 mg, 156.84 pmol, 50.04% yield, 95.88% purity, TFA salt) as a pale yellow gum.

[0442] 'll NMR (METHANOL-t / 4, 400 MHz): 5 ppm 8.27 - 8.19 (m, 1H), 7.94 - 7.84 (m, 1H), 7.79 (dd, J = 3.3, 8.1 Hz, 1H), 7.63 - 7.56 (m, 1H), 7.56 - 7.49 (m, 1H), 7.37 (dt, J = 1.4, 7.5 Hz, 1H), 7.23 (d, J = 8.4 Hz, 1H), 7.18 - 7.10 (m, 1H), 4.21 - 4.05 (m, 2H), 3.99 -3.75 (m, 1H), 3.41 - 3.33 (m, 1H), 3.25 - 3.17 (m, 1H), 3.12 (br d, J = 11.4 Hz, 1H), 3.05 -2A2.94 (m, 2H), 2.94 - 2.84 (m, 1H), 2.79 - 2.68 (m, 1H), 2.66 - 2.53 (m, 1H), 1.22 (dt, J = 2.9, 6.9 Hz, 3H), 0.77 (dd, J = 1.9, 6.4 Hz, 3H).

[0443] ¹³C NMR (METHANOL-d4, 101 MHz): 5 ppm 162.3, 162.0, 154.9, 153.9, 146.6, 134.9, 134.8, 130.9, 130.8, 129.3, 127.4, 127.3, 126.7, 126.5, 126.4, 120.7, 113.0, 112.9, 64.2, 64.1, 52.8, 52.6, 48.5, 43.1, 13.9, 13.8, 13.6, 13.5.

[0444] LCMS (ESI+): m / z 379.2 (M+H).

[0445] HRMS (TOF MS ES+): calcd for C22H27N4O2 [M+H]+: 379.2134 Found 379.2119.CW-9 & CW-10HK2CO3, DMF2 CW-9 CW-10

[0446] To a solution of Compound 2 (100 mg, 317.70 pmol) and Compound 2A (27.36 mg, 317.70 pmol) in DMF (2 mL) was added K2CO3 (131.72 mg, 953.09 pmol). The mixture was stirred at 80°C for 2 hrs. LC-MS showed Compound 2 was consumed and 29% of desired compound was detected. The reaction mixture was filtered. The residue was purified by Prep-HPLC (column: Phenomenex Luna C18 100*30mm*5 pm; mobile phase:[H2O (0.1% TFA) - ACN]: gradient: 5%-38% B over 8.0 min). The residue was purified by SFC (column: REGIS (s, s) WHELK-01 (250mm*30mm, 5um); mobile phase: [CO2-E1OH (0.1% NH3H2O)]; B%:50%, isocratic elution mode) to afford CW-10 (15.2 mg, 41.25 pmol, 75.17% yield, 98.91% purity) as a pale yellow solid and CW-9 (13.9 mg, 37.63 pmol, 68.56% yield, 98.65% purity) as a pale yellow solid.CW-9 spectra

[0447] ’ll NMR (METHANOL-74, 400 MHz): 5 ppm 8.23 (dd, 7=8.00, 1.00 Hz, 1 H) 7.84 - 7.95 (m, 1 H) 7.77 (d, 7=7.75 Hz, 1 H) 7.54 - 7.63 (m, 1 H) 7.45 - 7.53 (m, 1 H) 7.31 -7.40 (m, 1 H) 7.15 - 7.24 (m, 1 H) 7.11 (td, 7=7.63, 1.13 Hz, 1 H) 3.95 - 4.24 (m, 2 H) 3.32 -3.38 (m, 1 H) 3.19 - 3.26 (m, 1 H) 2.54 - 2.81 (m, 4 H) 2.14 - 2.41 (m, 4 H) 1.20 (1, 7=6.94 Hz, 3 H)

[0448] LCMS (ESI+): m / z 365.2 (M+H)Specific Rotation: [a]20°c589 nm = -80.2° ± 3.6 (c = 0.0640 g / 100 mL, methanol)CW-10 spectra

[0449] ’ll NMR (METHANOL-74, 400 MHz): 5 ppm 8.18 - 8.28 (m, 1 H) 7.83 - 7.92 (m, 1 H) 7.73 - 7.80 (m, 1 H) 7.55 - 7.61 (m, 1 H) 7.45 - 7.54 (m, 1 H) 7.37 (dd, J=7.69, 1.56 Hz, 1 H) 7.19 (d, J=8.25 Hz, 1 H) 7.11 (t, J=7.63 Hz, 1 H) 4.03 - 4.16 (m, 2 H) 3.33 - 3.38 (m, 1 H) 3.20 - 3.25 (m, 1 H) 2.63 - 2.75 (m, 4 H) 2.16 - 2.39 (m, 4 H) 1.20 (t, J=7.00 Hz, 3 H)

[0450] ¹³C NMR (METHANOL-d4, 101 MHz) 5 ppm 162.4, 154.3, 146.9, 134.7, 130.6, 129.8, 127.1, 126.7, 126.3, 125.3, 120.7, 120.2, 112.5, 63.9, 52.9, 48.2, 44.6, 13.6

[0451] LCMS (ES1+): mlz 365.2 (M+H)HRMS (TOF MS ES+): calcd for C21H25N4O2 [M+H]+: 365.1977 Found 365.1972.Specific Rotation: [a]20°c589 nm = +83.2° + 2.3 (c = 0.1010 g / 100 mL, methanol)CW-11 and CW-12

[0452] To a solution of Compound 1 (300 mg, 1.39 mmol) and Compound 1A (296.59 mg, 1.39 mmol) in ACN (5 mL) was added POC13 (1.06 g, 6.94 mmol, 647.08 pL). The mixture was stirred at 80°C for 12 hrs. LC-MS showed Compound 1 was consumed and 23% of desired compound was detected. The reaction mixture was diluted with NaHCO3 (10 mL) and extracted with EtOAc (10 mL * 3). The combined organic layers were washed with brine (10 mL * 2), dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 10 g ScpaFlash® Silica Flash Column, Eluent of 15% Ethyl acctatc / Pctrolcum ether gradient @ 80 mL / min) to afford Compound 2 (280 mg, 711.27 pmol, 51.23% yield) as a yellow solid.MS-ESI (m / z) calcd for CnHuBrC Ch [M+H]+: 395.0 / 393.0 Found 395.1 / 393.1. To a solution of Compound 2 (280 mg, 711.27 pmol) and Compound 2A (122.53 mg, 1.42 mmol) in DMF (4 mL) was added K2CO3 (294.90 mg, 2.13 mmol). The mixture was stirred at 80°C for 1 hr. LC-MS showed Compound 2 was consumed and 40% of desired compound was detected. The reaction mixture was filtered. The residue was purified by Prep-HPLC (column: Phenomenex Euna Cl 8 100*30mm*5 pm; mobile phase: [H2O (0.1% TFA) -ACN]; gradient: 10% - 45% B over 8.0 min) to afford Compound 3 (270 mg, 484.43 pmol, 68.11% yield, 100% purity) as a white solid. MS-ESI (m / z) calcd for C2iH23BrN4C>2 [M+H]+: 443.1 / 445.1 Found 443.2 / 445.2. Compound 3 (200 mg) was separated by SFC (column: REGIS (s,s) WHELK-01 (250mm*30mm, 5um); mobile phase: [CO2 - EtOH (0.1% NH3H2O)]; B%: 50%, isocratic elution mode) to afford CW-11 (73.56 mg, 165.66 pmol, 36.72% yield, 99.84% purity) as a white solid.

[0453] 1H NMR (METHANOL-t / 4400MHz): 5 ppm 8.22 (d, J=8.00 Hz, 1 H) 7.84 -7.93 (m, 1 H) 7.76 (d,.1=8.13 Hz, 1 H) 7.58 (t, J=7.57 Hz, 1 H) 7.36 - 7.39 (m, 1 H) 7.35 (br d, J=1.75 Hz, 2 H) 3.99 - 4.19 (m, 2 H) 3.37 (d,.1=13.88 Hz, 1 H) 3.14 - 3.28 (m, 1 H) 2.67 (t, J=4.82 Hz, 4 H) 2.15 - 2.35 (m, 4 H) 1.15 - 1.23 (m, 3 H).

[0454] LCMS (ESI+): mlz 443.2 / 445.2 (M+H)

[0455] CW-12 (62.49 mg, 140.67 pmol, 31.18% yield, 99.80% purity) as a white solid.

[0456] 1H NMR (METHANOL-r / 4400MHz): 5 ppm 8.15 - 8.26 (m, 1 H) 7.82 - 7.93 (m, 1 H) 7.76 (d, J=8.13 Hz, 1 H) 7.58 (td, J=7.57, 1.25 Hz, 1 H) 7.34 - 7.39 (m, 1 H) 7.24 -7.34 (m, 2 II) 4.01 - 4.17 (m, 2 II) 3.35 - 3.40 (m, 1 II) 3.17 - 3.26 (m, 1 II) 2.55 - 2.76 (m, 4 H) 2.17 - 2.35 (m, 4 H) 1.20 (t, J=6.94 Hz, 3 H)

[0457] LCMS (ESI+): mlz 443.2 / 445.2 (M+H).

[0458] The absolute configuration of CW-11 and CW-12 is uncertain.CW-13TFA, DCM K2CO3, DMF2 3 CW-13

[0459] To a solution of Compound 2 ( 100 mg, 317.70 pmol) and Compound 2A (68.08 mg, 317.70 pmol) in DMF (2 mL) was added K2CO3 (131.72 mg, 953.09 pmol). The mixture was stirred at 80°C for 12 hrs. LC-MS showed Compound 2 was consumed and 25% of desired compound was detected. The reaction mixture was diluted with H2O 5 mL and extracted with EtOAc (5 mL * 3). The combined organic layers were washed with brine (5 mL * 2), dried over anhydrous Na2SC>4, 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 20% Ethyl acetate / Petroleum ether gradient @ 40 mL / min) to afford Compound 3 (70 mg, 142.10 pmol, 44.73% yield) as a pale yellow oil. MS-ESI (m / z) calcd for C28H36N4O4 [M+H]+: 493.3 Found 493.33.

[0460] To a solution of Compound 3 (70 mg, 142.10 pmol) in DCM (1 mL) was added TFA (614.00 mg, 5.38 mmol, 0.4 mL). The mixture was stirred at 20°C for 1 hr. LC-MS showed Compound 3 was consumed and 76% of desired compound was detected. The reaction mixture was filtered and the filtrate was concentrated under vacuum. The residue was purified by Prep-HPLC (column: Phenomenex Luna C18 100*30mm*5 pm; mobile phase: [H2O (0.1% TFA) - ACN]; gradient: 10% - 40% B over 8.0 min) to afford CW-13 (34.31 mg, 66.55 pinol. 46.83% yield, 98.24% purity, TFA salt) as a colorless gum.

[0461] 'll NMR (METHANOL-d4, 400 MHz): 5 ppm 8.23 (d, J=8.00 Hz, 1 H), 7.83 -7.94 (m, 1 H), 7.78 (br d, J=8.38 Hz, 1 H), 7.49 - 7.62 (m, 2 H), 7.37 (ddd, J=7.69, 4.57, 1.63 Hz, 1 H), 7.24 (d, J=8.25 Hz, 1 H), 7.15 (tdd, J=7.61, 7.61, 2.22, 1.06 Hz, 1 H), 4.01 - 4.22 (m, 2 H), 3.67 - 3.88 (m, 1 H), 3.35 - 3.55 (m, 1 H), 3.10 - 3.21 (m, 2 H), 2.88 - 3.07 (m, 2 H), 2.53 - 2.86 (m, 3 H), 1.41 - 1.58 (m, 1 H), 1.10 - 1.30 (m, 4 H), 0.68 - 0.81 (m, 3 H)

[0462] LCMS (FSI+): m / z 393.1 (M+H).CW-14TFA, DCM K2CO3, Nal, ACNCW-14

[0463] To a solution of Compound 2 (80 mg, 254.16 pmol) and Compound 2A (69.64 mg, 304.99 pmol) in ACN (1 mL) was added Nal (7.62 mg, 50.83 pmol) and K2CO3 (105.38 mg, 762.47 pmol). The mixture was stirred at 80°C for 12 hrs. LC-MS showed Compound 2 was consumed and 67% of desired compound was detected. The reaction mixture was diluted with H2O 5 mL and extracted with EtOAc (5 mL * 3), dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to afford Compound 3 (100 mg, crude) as a yellow oil. MS-ESI (m / z) calcd for C29H38N4O4 [M+H]+: 507.3 Found 507.3.

[0464] To a solution of Compound 3A (100 mg, 197.38 pmol) in DCM (1 mL) was added TFA (767.50 mg, 6.73 mmol, 0.5 mL). The mixture was stirred at 20°C for 2 hrs. LC-MS showed Compound 3A was consumed and 68% of desired compound was detected. The reaction mixture was filtered. The residue was purified by Prep-HPLC (column:Phenomenex Luna C18 100*30mm*5 pm; mobile phase: [H2O (0.1% TFA) - ACN]; gradient: 5% - 40% B over 8.0 min) to afford CW-14 (86.79 mg, 166.73 pmol, 84.47% yield, 100% purity, TFA salt) as a pale yellow gum.

[0465] 'll NMR (METHANOL-t / 4, 400 MHz): 5 ppm 8.22 (d, J=7.88 Hz, 1 H), 7.83 -7.96 (m, 1 H), 7.77 (d, J=8.00 Hz, 1 H), 7.48 - 7.63 (m, 2 H), 7.35 (ddd, J=17.48, 7.72, 1.31 Hz, 1 H), 7.24 (dd, J=8.25, 4.25 Hz, 1 H), 7.15 (td, J=7.54, 3.44 Hz, 1 H), 3.99 - 4.21 (m, 2 H), 3.44 - 3.82 (m, 2 H), 3.33 - 3.42 (m, 1 H), 3.12 - 3.29 (m, 2 H), 2.55 - 3.04 (m, 4 H), 2.10 - 2.32 (m, 1 H), 1.22 (q, J=6.96 Hz, 3 H), 0.91 (dd,.1=6.88, 3.00 Hz, 3 H), 0.65 (dd, J=14.38, 6.88 Hz, 3 H)

[0466] LCMS (ESI+): m / z 407.2 (M+H).CW-15TFA, DCM

[0467] To a solution of Compound 2 (90 mg, 285.93 pmol) and Compound 2A (73.53 mg, 343.11 pmol) in ACN (2 mL) was added Nal (8.57 mg, 57.19 pmol) and K2CO3 (118.55 mg, 857.78 pmol). The mixture was stirred at 80°C for 12 hrs. LC-MS showed Compound 2 was consumed and 54% of desired compound was detected. The reaction mixture was diluted with H2O 10 mL and extracted with EtOAc (10 mL * 3), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to afford Compound 4 (100 mg, crude) as a yellow oil. MS-ESI (m / z) calcd for C28H36N4O4 [M+H]+: 493.3 Found 493.3.

[0468] To a solution of Compound 4 (100 mg, 203.00 pmol) in DCM (1 mL) was added TEA (307.00 mg, 2.69 mmol, 0.2 mL). The mixture was stirred at 20°C for 3 hrs. LC-MS showed 19% of Compound 4 was remained and 35% of desired compound was detected. The reaction mixture was filtered and the filtrate was concentrated under vacuum. The residue was purified by Prep-HPLC (column: Phenomenex Luna C18 100*30mm*5 pm; mobile phase: [H2O (0.1% TFA) - ACN]; gradient: 5% - 40% B over 8.0 min) to afford CW-15 (21.7 mg, 42.50 pmol, 20.94% yield, 99.20% purity, TFA salt) as a pale yellow gum.

[0469] ’ll NMR (METHANOL-r / 4, 400 MHz): 5 ppm 8.22 (d, J=7.13 Hz, 1 H), 7.84 -7.92 (m, 1 H), 7.74 - 7.83 (m, 1 H), 7.50 - 7.61 (m, 2 H), 7.33 (br d, J=7.75 Hz, 1 H), 7.26 (d,.1=8.25 Hz, 1 H), 7.16 (t,.1=7.57 Hz, 1 H), 4.03 - 4.19 (m, 2 H), 3.88 - 4.02 (m, 2 H), 3.62 -3.87 (m, 2 II), 3.17 - 3.29 (m, 2 II), 2.67 - 2.83 (m, 2 II), 1.22 (t,.1=6.94 Hz, 3 II), 1.05 - 1.17 (m, 6 H)

[0470] LCMS (ESI+): mlz 393.2 (M+H).CW-16BocH TFA, DCM K2CO3, Nal, ACN2 3 CW-16

[0471] To a solution of Compound 2 (80 mg, 254.16 pmol) and Compound 2A (65.36 mg, 304.99 pmol) in ACN (1 mL) was added Nal (7.62 mg, 50.83 pmol) and K2CO3 (105.38 mg, 762.47 pmol). The mixture was stirred at 80°C for 12 hrs. LC-MS showed Compound 2 was consumed and 73% of desired compound was detected. The reaction mixture was diluted with H2O 10 mL and extracted with EtOAc (10 mL * 3), dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to afford Compound 3 (100 mg, crude) as a yellow oil. MS-ESI (m / z) calcd for C28H36N4O4 [M+H]+: 493.3 Found 493.3.

[0472] To a solution of Compound 3 (100 mg, 203.00 pmol) in DCM (1 mL) was added TFA (767.50 mg, 6.73 mmol, 0.5 mL). The mixture was stirred at 20°C for 2 hrs. LC-MS showed Compound 3 was consumed and 70% of desired compound was detected. The reaction mixture was filtered. The filtrate was purified by Prep-HPLC (column: Phenomenex Luna C18 100*30mm*5 μm; mobile phase: [H2O (0.1% TFA) - ACN]; gradient: 5% - 40% B over 8.0 min) to afford CW-16 (44.16 mg, 87.10 pmol, 42.90% yield, 99.90% purity, TFA salt) as a pale yellow gum.

[0473] ’ll NMR (METHANOL- 4, 400 MHz): 5 ppm 8.23 (br d, J=7.88 Hz, 1 H), 7.84 - 7.94 (m, 1 H), 7.71 - 7.83 (m, 1 H), 7.59 (td, J=7.35, 3.94 Hz, 1 H), 7.47 - 7.56 (m, 1 H), 7.33 - 7.43 (m, 1 H), 7.23 (d, J=8.38 Hz, 1 H), 7.15 (t, J=7.57 Hz, 1 H), 4.00 - 4.25 (m, 2 H), 3.77 - 3.98 (m, 1 H), 3.77 (br s, 1 H), 2.88 - 3.13 (m, 4 H), 2.75 (br d, J=6.75 Hz, 2 H), 1.22 (dt, J=9.60, 7.02 Hz, 3 H), 0.92 (br d, J=5.75 Hz, 6 H)

[0474] LCMS (ESI+): m / z 393.2 (M+H).CW-17K2CO3, Nal. ACN2

[0475] To a solution of Compound 2 (60 mg, 190.62 pmol) and Compound 2A (26.12 mg, 228.74 pmol) in ACN (1 mL) was added K2CO3 (79.03 mg, 571.85 pmol) and Nal (5.71 mg, 38.12 pmol). The mixture was stirred at 80°C for 1 hr. LC-MS and HPLC showed Compound 2 was consumed and 42% of desired compound was detected. The reaction mixture was filtered. The residue was purified by Prep - HPLC (TFA condition; column: Phenomenex Luna C18 100*30mm*5 pm; mobile phase: [H2O (0.1% TFA) - ACN]; gradient: 10% - 40% B over 8.0 min) to afford CW-17 (33.21 mg, 65.26 pmol, 34.23% yield, 99.53% purity, TFA) as a white solid.

[0476] 1H NMR (METHANOL-^4400MHz): 5 ppm 8.27 - 8.19 (m, 1H), 7.93 - 7.85 (m, 1H), 7.78 (br d, J = 8.3 Hz, 1H), 7.63 - 7.48 (m, 2H), 7.44 - 7.32 (m, 1H), 7.26 - 7.19 (m, 1H), 7.18 - 7.10 (m, 1H), 4.21 - 4.03 (m, 2H), 4.00 - 3.78 (m, 1H), 3.65 - 3.37 (m, 1H), 3.26 (br d, J = 14.8 Hz, 1H), 3.17 - 3.11 (m, 1H), 2.99 - 2.65 (m, 2H), 2.60 - 2.38 (m, 2H), 1.36 -1.10 (m, 6H), 1.03 - 0.65 (m, 3H)

[0477] LCMS (ESI+): m / z 393.2 (M+H).CW-18Boc TFA, DCM K2CO3, Nal, DMF2 3

[0478] To a solution of Compound 2 (100 mg, 317.70 pmol) and Compound 2A (80.93 mg, 381.24 pmol) in DMF (2 mL) was added K2CO3 (131.73 mg, 953.09 pmol) and Nal (4.76 mg, 31.77 pmol). The mixture was stirred at 50°C for 1 hr. LC-MS showed Compound 2 was consumed and 33% of desired compound was detected. The reaction mixture was diluted with H2O (2 mL) and extracted with EtOAc (2 mL * 3). The combined organic layers were washed with brine (3 mL * 1), dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to afford Compound 5 (120 mg, crude) as a yellow oil. MS-ESI (m / z) calcd for C28H34N4O4 [M+H]+: 491.3 Found 491.3. To a solution of Compound 3 (120 mg, 244.60 pmol) in DCM (1 mL) was added TFA (557.79 mg, 4.89 mmol, 363.38 pL). The mixture was stirred at 20°C for 1 hr. LC-MS and HPLC showed Compound 3 was consumed and 53% of desired compound was detected. The reaction mixture was concentrated under vacuum. The residue was purified by Prep-HPLC (TFA condition; column: Phenomenex Luna Cl 8 100*30mm*5um; mobile phase: [H2O (0.1% TFA) - ACN]; gradient: 5% - 40% B over 8.0 min) to afford CW-18 (46.04 mg, 86.26 pmol, 35.26% yield, 94.52% purity, TFA) as a yellow solid.Spectrum:

[0479] 1H NMR (METHANOL-^4400MHz): 5 ppm 8.22 (d, J = 8.0 Hz, 1H), 7.94 -7.86 (m, 1H), 7.77 (d, J = 8.1 Hz, 1H), 7.60 (t, J = 7.6 Hz, 1H), 7.56 - 7.49 (m, 1H), 7.27 (dd, J = 1.4, 7.6 Hz, 1H), 7.17 (d, J = 8.4 Hz, 1H), 7.13 - 7.07 (m, 1H), 4.06 (q, J = 6.9 Hz, 2H), 3.92 - 3.72 (m. 2H), 3.26 - 3.15 (m, 2H), 3.07 - 2.87 (m, 4H), 1.21 (t, J = 7.0 Hz, 3H), 0.44 (br s, 2H), 0.05 (br s, 2H)

[0480] LCMS (ESI+): m / z 391.2 (M+H).CW-19

[0481] To a solution of Compound 1 (500 mg, 3.31 mmol) and Compound 1A (410.94 mg, 3.64 mmol, 289.80 pL) in THE (5 mL) was added K2CO3 (1.37 g, 9.92 mmol). The mixture was stirred at 70°C for 1 hr. LC-MS showed Compound 1 was consumed and 72% of desired compound was detected. The reaction mixture was filtered and concentrated under reduced pressure to remove solvent to afford Compound 2 (1 g, crude) as a black solid. MS-ESI (m / z) calcd for C10H10ClNO3 [M+H]+: 228.0 / 230.0 Found 228.1 / 230.1.

[0482] To a solution of Compound 2 (500 mg, 2.20 mmol) and Compound 2A (301.30 mg, 2.20 mmol, 287.50 pL) in ACN (10 mL) was added POC13 (1.01 g, 6.59 mmol, 614.19 pL). The mixture was stirred at 80°C for 12 hrs. LC-MS showed Compound 2 was consumed and 34% of desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The reaction mixture was diluted with saturated NaHCO3 solution 20 mL and extracted with EtOAc (20 mL * 3), the combined organic layer was dried over anhydrous Na2SC>4, filtered and concentrated 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 10% Ethyl acetate / Petroleum ether gradient @ 80 mL / min) to afford Compound 3 (400 mg, 1.22 mmol, 55.39% yield) as a yellow solid. MS-ESI (m / z) calcd for C18H17ClN2O2 [M+H]+:329.1 / 331.1 Found 328.9 / 330.9.

[0483] To a solution of Compound 3 (150 mg, 456.22 pmol) in DMF (1 mL) was added K2CO3 (189.16 mg, 1.37 mmol) and Compound 3A (78.59 mg, 912.43 pmol). The mixture was stirred at 80°C for 1 hr. LC-MS showed Compound 3 was consumed and 32% of desired compound was detected. The reaction mixture was filtered. The filtrate was purified by Prep-HPLC (column: Phenomenex Luna C18 100*40mm*5 μm; mobile phase: [H2O (0.1% TFA) - ACN]; gradient: 10%-40% B over 8.0 min) to afford CW-19 (143.55 mg, 291.30 pmol, 63.85% yield, 99.94% purity, TFA salt) as a white solid.

[0484] ’ll NMR (METHANOL-d4, 400 MHz): 5 ppm 8.02 - 8.10 (m, 1 H), 7.73 (d, J=6.75 Hz, 1 H), 7.49 - 7.57 (m, 1 H), 7.45 (t,.1=7.63 Hz, 1 H), 7.39 (dd,.1=7.75, 1.63 Hz, 1 H), 7.23 (d, J=8.25 Hz, 1 H), 7.14 (td, J=7.60, 1.06 Hz, 1 H), 4.02 - 4.21 (m, 2 H), 3.44 - 3.59 (m, 2 H), 3.08 - 3.21 (m, 4 H), 2.67 - 2.94 (m, 4 H), 2.61 - 2.67 (m, 3 H), 1.21 (t, J=7.00 Hz, 3 H)

[0485] LCMS (ESI+): m / z 379.2 (M+H).

[0486] To a solution of Compound 1 (1 g, 6.62 mmol) and Compound 1A (821.88 mg, 7.28 mmol, 579.60 pL) in THF (20 mL) was added K2CO3 (2.74 g, 19.85 mmol). The mixture was stirred at 70°C for 1 hr. LC-MS showed Compound 1 was consumed and 96% of desired compound was detected. The reaction mixture was filtered and the filtrate was dried under vacuum to afford Compound 2 (1.42 g, crude) as a yellow solid. MS-ESI (m / z) calcd for C10H10CINO3 [M+H]+: 228.0 / 230.0 Found 227.8 / 229.6.

[0487] To a solution of Compound 2 (700 mg, 3.07 mmol) and Compound 2A (421.82 mg, 3.07 mmol, 402.50 pL) in ACN (20 mL) was added POC13 (1.41 g, 9.22 mmol, 859.87 pL). The mixture was stirred at 80°C for 12 hrs. LC-MS showed Compound 2 was consumed and 37% of desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with EtOAc 10 mL and then was poured into saturated Na2CO3 solution to pH = 8 - 9, the mixture was extracted with EtOAc (10 mL * 3). The combined organic layers were 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 Flash Column, Eluent of 0-10% Ethyl acetate / Petroleum ether gradient @80 mL / min) to afford Compound 3 (500 mg, 1.49 mmol, 48.47% yield, 98% purity) as a yellow solid. MS-ESI (m / z) calcd for C18H17CIN2O2 [M+H]+:329.1 / 331.0 Found 329.1 / 331.1.

[0488] To a solution of Compound 3 (200 mg, 608.29 pmol) and Compound 3A (62.87 mg, 729.94 pmol) in DMF (2 mL) was added K2CO3 (252.21 mg, 1.82 mmol). The mixture was stirred at 80°C for 1 hr. LC-MS and HPLC showed Compound 3 was consumed and 27% of desired compound was detected. The reaction mixture was filtered. The filtrate was purified by Prep-HPLC (TFA condition; column: Phenomenex Luna C18 100*40mm*5 pm; mobile phase: [H2O (0.1 % TFA) - ACN]; gradient: 10% - 40% B over 8.0 min) to afford CW-20 (99.35 mg, 197.33 pmol, 32.44% yield, 97.82% purity, TFA salt) as a white solid.

[0489] NMR (METHAN0W4, 400 MHz): 5 ppm 8.12 (d, J = 8.1 Hz, 1H), 7.59 (s, 1H), 7.56 - 7.48 (m, 1H), 7.44 (d,.1 = 8.3 Hz, 1H), 7.39 (dd, J = 1.4, 7.7 Hz, 1H), 7.22 (d, J = 7.8 Hz, 1H), 7.14 (dt,.1 = 0.9, 7.6 Hz, 1H), 4.24 - 3.98 (m, 2H), 3.52 - 3.37 (m, 2H), 3.10 (br d, J = 4.6 Hz, 4H), 2.67 (br dd, J = 4.6, 12.2 Hz, 2H), 2.55 (s, 5H), 1.21 (t, J = 7.0 Hz, 3H)

[0490] LCMS (ESI+): m / z 379.2 (M+H).CW-21

[0491] To a solution of Compound 1 ( 1 g, 6.62 mmol) in THF (20 mL) was added K2CO3 (2.74 g, 19.85 mmol) was added Compound 1A (821.88 mg, 7.28 mmol, 579.60 pL) at 20°C. The mixture was stirred at 70°C for 1 hr. LC-MS showed Compound 1 was consumed and 97% of desired compound was detected. The reaction mixture was filtered to afford Compound 2 (1.4 g, crude) as a yellow solid. MS-ESI (m / z) calcd for C10H10C1NO3 [M+H]+:228.0 / 230.0 Found 227.8 / 230.0.

[0492] To a solution of Compound 2 (700 mg, 3.07 mmol) and Compound 2A (464.00 mg, 3.38 mmol, 442.75 pL) in ACN (10 mL) was added POC13 (1.41 g, 9.22 mmol, 859.87 pL). The mixture was stirred at 80°C for 7 hrs. LC-MS showed Compound 2 was consumed and 39% of desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with EtOAc 10 mL and then was poured into saturated Na2CO3 solution to pH = 8 ~ 9, the mixture was extracted withEtOAc (10 mL * 3). The combined organic layers were 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 0-10% Ethyl acetate / Petroleum ether gradient @ 80 mL / min) to afford Compound 3 (450 mg, 1.34 mmol, 43.62% yield, 98% purity) as a yellow solid. MS-ESI (m / z) calcd for C18H17C1N2O2 [M+H]+: 329.1 / 331.0 Found 329.1 / 331.0.

[0493] To a solution of Compound 3 (150 mg, 456.22 pmol) and Compound 3A (47.16 mg, 547.46 pmol) in DMF (2 mL) was added K2CO3 (189.15 mg, 1.37 mmol). The mixture was stirred at 80°C for 1 hr. LC-MS showed Compound 3 was consumed and 64% of desired compound was detected. The reaction mixture was filtered. The filtrate was purified by Prep-HPLC (TFA condition; column: Phenomenex Luna C18 100*40mm*5 pm; mobile phase: [H2O (0.1% TFA) - ACN]; gradient; 10% - 40% B over 8.0 min) to afford CW-21 (123.62 mg, 250.63 pmol, 54.94% yield, 99.85% purity, TFA salt) as a white solid.

[0494] ’ll NMR (METHANOL-r / 4, 400 MHz): 5 ppm 8.03 (s, 1H), 7.79 - 7.66 (m, 2H), 7.53 (t, J = 7.9 Hz, 1H), 7.40 (dd, J = 1.3, 7.8 Hz, 1H), 7.23 (d, J = 8.4 Hz, 1H), 7.14 (t, J = 7.5 Hz, 1H), 4.29 - 3.95 (m, 2H), 3.61 - 3.41 (m, 2H), 3.22 - 3.08 (m, 4H), 2.86 - 2.57 (m, 4H), 2.51 (s, 3H), 1.21 (t, J = 7.0 Hz, 3H)

[0495] LCMS (ESI+): m / z 379.2 (M+H).CW-22

[0496] To a solution of Compound 1 (500 mg, 3.31 mmol) and Compound 1A (410.94 mg, 3.64 mmol, 289.80 pL) in THF (5 mL) was added K2CO3 (1.37 g, 9.92 mmol). The mixture was stirred at 70°C for 1 hr. LC-MS showed Compound 1 was consumed and 92% of desired compound was detected. The reaction mixture was filtered and concentrated under reduced pressure to afford Compound 2 (1 g, crude) as a white solid. MS-ESI (m / z) calcd for C10H10CINO [M+H]+: 228.0 / 230.0 Found 228.2 / 230.2.

[0497] To a solution of Compound 2 (500 mg, 2.20 mmol) and Compound 2A (301.30 mg, 2.20 mmol, 287.50 pL) in ACN (10 mL) was added POC13 (1.01 g, 6.59 mmol, 614.18 pL). The mixture was stirred at 80°C for 12 hrs. LC-MS showed Compound 2 was consumed and 54% of desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The reaction mixture was diluted with saturated NaHCO3 solution 20 mL and extracted with EtOAc (20 mL * 3), the combined organic layer was dried over anhydrous 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 Flash Column, Eluent of 10% Ethyl acetate / Petroleum ether gradient @ 80 mL / min) to afford Compound 3 (100 mg, 304.14 pmol, 13.85% yield) as a white solid. MS-ESI (m / z) calcd for C18H17C1N2O2 [M+H]+: 329.1 / 331.0 Found 328.9 / 330.9.

[0498] To a solution of Compound 3 (100 mg, 304.14 pmol) in DMF (2 mL) was added K2CO3 (126.10 mg, 912.43 pmol) and Compound 3A (52.39 mg, 608.29 pmol). The mixture was stirred at 80°C for 1 hr. LC-MS showed Compound 3 was consumed and 33% of desired compound was detected. The reaction mixture was filtered. The filtrate was purified by Prep-HPLC (column: Phenomenex Luna C18 100*40mm* 5 pm; mobile phase: [H2O (0.1% TFA) - ACN]; gradient: 10% - 40% B over 8.0 min) to afford CW-22 (49.06 mg, 99.48 pmol, 32.71% yield, 99.86% purity, TFA salt) as a white solid.

[0499] ’ll NMR (METHANOL-r / 4, 400 MHz): 5 ppm 7.66 - 7.79 (m, 1 H), 7.57 - 7.64 (m, 1 II), 7.48 - 7.55 (m, 1 II), 7.33 - 7.43 (m, 2 II), 7.23 (d, 8.25 IIz, 1 II), 7.14 (td, J=7.60, 1.06 Hz, 1 H), 4.04 - 4.22 (m, 2 H), 3.41 - 3.51 (m, 2 H), 3.13 (t, J=5.13 Hz, 4 H), 2.80 (s, 3 H), 2.67 - 2.77 (m, 2 H), 2.55 - 2.65 (m, 2 H), 1.23 (t, J=6.94 Hz, 3 H).

[0500] LCMS (ESI+): m / z 379.2 (M+H).CW-23

[0501] To a solution of Compound 1A (900 mg, 5.88 mmol) and EtI (1.10 g, 7.05 mmol, 564.08 pL) in DMF (20 mL) was added K2CO3 (2.44 g, 17.63 mmol). The mixture was stirred at 80°C for 2 hrs. TLC (SiO2, Petroleum ether: Ethyl acetate=5:l, Rf (Pl) =0.43) indicated Compound 1 A was consumed and one major new spot with larger polarity was detected. The reaction mixture was diluted with H2O 20 mL and extracted with EtOAc (20 ml, * 3). The combined organic layers were washed with brine (50 mL * 1 ), dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to afford Compound 2A (1 g, crude) as a yellow oil.

[0502] To a solution of Compound 2A (1.10 g, 6.07 mmol) in EtOH (40 mL) and H2O (10 mL) was added Fe (1.02 g, 18.21 mmol) and NH4CI (1.62 g, 30.36 mmol). The mixture was stirred at 80°C for 1 hr. TLC (SiO2, Petroleum ether: Ethyl acetate=5:l, Rf (Pl) = 0.41) indicated Compound 2A was consumed and one major new spot with larger polarity was detected. The reaction mixture was filtered and then the mixture was poured into saturated NaHCO3 solution to pH = 8 ~ 9, then the mixture was extracted with EtOAc (10 mL * 3). The combined organic layers were dried over anhydrous Na2SO4 filtered and concentrated under reduced pressure to give a residue to afford Compound 3 A (600 mg, crude) as a yellow oil.

[0503] ’ll NMR (DMSO-d6, 400 MHz): 6 ppm 6.73 - 6.63 (m, 1H), 6.55 - 6.48 (m, 1H), 6.35 (d, J = 7.4 Hz, 1H), 4.70 (s, 2H), 3.76 (q, J = 7.0 Hz, 2H), 2.13 (s, 3H), 1.31 (t, J = 7.0 Hz, 3H)

[0504] MS -ESI (m / z) calcd for C9H13NO [M+H]+: 152.1 Found 152.5.

[0505] To a solution of Compound 3A (150 mg, 702.19 pmol) and Compound 3B (106.17 mg, 702.19 pmol) in ACN (3 mL) was added POCI3 (323.01 mg, 2.11 mmol, 196.36 pL). The mixture was stirred at 80°C for 12 hrs. LC-MS showed Compound 3A was consumed and 59% of desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with EtOAc 5 mL and then was poured into saturated Na2CO3 solution to pH = 8 ~ 9, the mixture was extracted with EtOAc (5 mL * 3). The combined organic layers were dried over Na2SO4, 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 0~8% Ethyl acetate / Petroleum ether gradient @ 60 mL / min) to afford Compound 4 A (140mg, 400.25 pmol, 57.00% yield, 94% purity) as a yellow solid. MS-ESI (m / z) calcd for C18H17CIN2O2 [M+H]+: 329.1 / 331.0 Found 329.1 / 331.2.

[0506] To a solution of Compound 4A (140 mg, 425.80 pmol) and Compound 4B (55.01 mg, 638.70 pmol) in DMF (2 mL) was added K2CO3 (176.54 mg, 1.28 mmol). The mixture was stirred at 80°C for 1 hr. LC-MS showed Compound 4A was consumed and 32% of desired compound was detected. The reaction mixture was filtered. The filtrate was purified by Prep-HPLC (TFA condition; column: Phenomenex Luna C18 100*30mm*5 pm; mobile phase: [H2O (0.1% TFA) - ACN]; gradient: 5% - 40% B over 8.0 min) to afford CW-23 (47.57 mg, 96.41 pmol, 22.64% yield, 99.81% purity, TFA salt) as a pale blue solid.

[0507] 'll NMR (METH ANOL-74, 400 MHz): 5 ppm 8.26 (dd, J = 0.8, 7.9 Hz, 1H), 7.96 - 7.86 (m, 1H), 7.81 (d, J = 8.1 Hz, 1H), 7.62 (t, J = 7.6 Hz, 1H), 7.46 - 7.36 (m, 1H), 7.30 - 7.18 (m, 2H), 4.06 - 3.86 (m, 1H), 3.74 - 3.58 (m, 1H), 3.47 (s, 2H), 3.20 - 3.03 (m, 4H), 2.76 - 2.64 (m, 2H), 2.61 - 2.47 (m, 2H), 2.40 (s, 3H), 1.01 (t, J = 7.1 Hz, 3H)

[0508] LCMS (ESI+): m / z 379.1 (M+H).CW-24

[0509] To a solution of Compound 1 (1 g, 6.53 mmol) and EtI (1.22 g, 7.84 mmol, 626.76 pL) in DMF (20 mL) was added K2CO3 (2.71 g, 19.59 mmol) at 20°C. The mixture was stirred at 80°C for 2 hrs. TLC (SiO2 Petroleum ether: Ethyl acetate=5:l, Rf (Pl) = 0.35, platel) indicated Compound 1 was consumed and one major new spot with larger polarity was detected. The reaction mixture was diluted with H2O 20 mL and extracted with EtOAc (20 mL * 3). The combined organic layers were washed with brine (50 mL * 1), dried over anhydrous Na2SO4 filtered and concentrated under reduced pressure to give a residue to afford Compound 2 (1.1 g, crude) as a yellow oil.

[0510] ’ll NMR (DMSO-76, 400 MHz): 8 ppm 7.76 (d, J = 8.3 Hz, 1H), 7.16 (s, 1H), 6.89 (dd, 7= 0.8, 8.3 Hz, 1H), 4.18 (q, J = 6.9 Hz, 2H), 2.37 (s, 3H), 1.33 (t, J = 7.0 Hz, 3H)

[0511] MS-ESI (m / z) calcd for C9H11NO3 [M+H]+: 182.0 Found 182.4.

[0512] To a solution of Compound 2 (1.1 g, 6.07 mmol) in EtOH (40 mL) and H2O ( 10 mL) was added Fe (1.02 g, 18.21 mmol) and NH4C1 (1.62 g, 30.36 mmol). The mixture was stirred at 80°C for 1 hr. TLC (S1O2, Petroleum ether: Ethyl acetate=5:l, Rf (Pl) = 0.43, plate 1) indicated Compound 2 was consumed and one major new spot with larger polarity was detected. The reaction mixture was filtered and then the mixture was poured into saturated NaHCO₃ solution to pH = 8~9, then the mixture was extracted with EtOAc (10 mL * 3). The combined organic layers were dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to afford Compound 3 (620 mg, crude) as a yellow oil.

[0513] ’ll NMR (DMSO-d6, 400 MHz): 5 ppm 6.59 (s, 1H), 6.54 - 6.49 (m, 1H), 6.49 -6.44 (m, 1H), 4.40 (br s, 2H), 3.96 (q, J = 6.9 Hz, 2H), 2.15 (s, 3H), 1.32 (t, J= 6.9 Hz, 3H)

[0514] MS-ESI (m / z) calcd for C9H13NO [M+H]+: 152.1 Found 152.5.

[0515] To a solution of Compound 3 (150 mg, 702.19 pmol) and Compound 3A (106.17 mg, 702.19 pmol) in ACN (3 mL) was added POC13 (323.01 mg, 2.11 mmol, 196.36 pL). The mixture was stirred at 80°C for 12 hrs. LC-MS showed Compound 3 was consumed and 61% of desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with EtOAc 5 mL and then was poured into saturated Na2CO3 solution to pH = 8~9, the mixture was extracted with EtOAc (5 mL * 3). The combined organic layers were dried over Na2SO4, 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 0~8% Ethyl acetate / Petroleum ether gradient @ 60 mL / min) (SiO2, Petroleum ether: Ethyl acetate=3:l, Rf (Pl) = 0.51, platel) to afford Compound 4 (140 mg, 417.28 pmol, 59.43% yield, 98% purity) as a yellow solid. MS-ESI (m / z) calcd for C18H17CIN2O2 [M+H]+:329.1 / 331.0 Found 329.0 / 331.0.

[0516] To a solution of Compound 4 (140 mg, 425.80 pmol) and Compound 4A (55.01 mg, 638.70 pmol) in DMF (2 mL) was added K2CO3 (176.54 mg, 1.28 mmol). The mixture was stirred at 80°C for 1 hr. LC-MS showed Compound 4 was consumed and 34% of desired compound was detected. The reaction mixture was filtered. The filtrate was purified by Prep-HPLC (TFA condition; column: Phenomenex Luna Cl 8 100*30mm*5 pm; mobile phase: [II2O (0.1% TFA) - ACN]; gradient: 5%-40% B over 8.0 min) to afford CW-24 (87.8 mg, 177.65 pmol, 41.72% yield, 99.65% purity, TFA salt) as a pale blue solid.

[0517] ’ll NMR (METHANOL-74, 400 MHz): 5 ppm 8.29 - 8.19 (m, 1H), 7.94 - 7.84 (m, 1H), 7.77 (d, 7= 8.0 Hz, 1H), 7.63 - 7.54 (m, 1H), 7.26 (d, 7 = 7.9 Hz, 1H), 7.05 (s, 1H), 6.96 (d, 7 = 7.8 Hz, 1H), 4.20 - 4.00 (m, 2H), 3.48 - 3.36 (m, 2H), 3.10 (t, 7= 5.1 Hz, 4H), 2.73 - 2.61 (m, 2H), 2.59 - 2.49 (m, 2H), 2.45 (s, 3H), 1.20 (t, 7= 6.9 Hz, 3H)

[0518] ¹³C NMR (METHANOL-d4, 101 MHz): 5 ppm 162.3, 153.8, 153.5, 146.6, 141.7, 134.8, 129.4, 127.3, 126.7, 126.4, 122.3, 121.1, 120.7, 113.6, 64.0, 49.0, 43.1, 20.4, 13.6.

[0519] LCMS (ESI+): m / z 379.2 (M+H).

[0520] HRMS (TOF MS+): calcd for C22H27N4O2 [M+H]+: 379.2134 Found 379.2119.CW-25

[0521] To a solution of Compound 1 (150 mg, 702.19 pmol) and Compound 1A (106.17 mg, 702.19 pmol) in ACN (3 mL) was added POCI3 (323.01 mg, 2.11 mmol, 196.36 pL). The mixture was stirred at 80°C for 12 hrs. LC-MS showed 11% of Compound 1 remained and 40% of desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The reaction mixture was diluted with H2O 10 mL and extracted with EtOAc (10 mL * 3), the combined organic layer was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 10 g SepaFlash® Silica Flash Column, Eluent of 10% Ethyl acetate / Petroleum ether gradient @ 60 mL / min) to afford Compound 2 (100 mg, 304.14 pmol, 43.31% yield) as a pale yellow solid. MS-ESI (m / z) calcd for C18H17CIN2O2 [M+H]+: 329.1 / 331.0 Found 329.1 / 331.1.

[0522] To a solution of Compound 2 (100 mg, 304.14 pmol) and Compound 2A (52.39 mg, 608.29 pmol) in DMF (2 mL) was added K2CO3 (126.10 mg, 912.43 pmol). The mixture was stirred at 80°C for 0.5 hr. LC-MS showed Compound 2 was consumed and 37%of desired compound was detected. The reaction mixture was filtered. The filtrate was purified by Prep-HPLC (column: Phenomenex Luna C18 100*30mm*5 pm; mobile phase:[H2O (0.1% TFA) - ACN]; gradient: 5% - 40% B over 8.0 min) to afford CW-25 (65.27 mg, 132.38 pmol, 43.53% yield, 99.89% purity, TFA salt) as a white solid.

[0523] ’ll NMR (METHANOL-r / 4, 400 MHz): 5 ppm 8.24 (d, J=7.88 Hz, 1 H), 7.84 -7.95 (m, 1 H), 7.75 - 7.82 (m, 1 H), 7.60 (t, J=7.57 Hz, 1 H), 7.33 (br d, J=8.38 Hz, 1 H), 7.22 (d, J=1.88 Hz, 1 H), 7.11 (d, J=8.38 Hz, 1 H), 3.98 - 4.17 (m, 2 H), 3.38 - 3.60 (m, 2 H), 3.00 - 3.18 (m, 4 H), 2.48 - 2.82 (m, 4 H), 2.38 (s, 3 H), 1.19 (t, J=7.00 Hz, 3 H)

[0524] LCMS (ESI+): m / z 379.2 (M+H).CW-26H POCI3, ACN K2CO3, DMF1A 2A CW-26

[0525] To a solution of Compound 1A (150.00 mg, 702.19 pmol) and Compound IB (106.17 mg, 702.19 pmol) in ACN (3 mL) was added POC13 (323.01 mg, 2.11 mmol, 196.36 pL). The mixture was stirred at 80°C for 12 hrs. LC-MS showed 11% of Compound 1 A was remained and 67% of desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The reaction mixture was diluted with H2O 10 mL and extracted with EtOAc (10 mL * 3), the combined organic layer was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 10 g SepaFlash® Silica Flash Column, Eluent of 10% Ethyl acetate / Petroleum ether gradient @ 80 mL / min) to afford Compound 2A (100 mg, 304.14 pmol, 43.31% yield) as a pale yellow solid. MS-ESI (m / z) calcd for C18H17CIN2O2 [M+H]+: 329.1 / 331.1 Found 329.1 / 331.2.

[0526] To a solution of Compound 2A ( 100 mg, 304.14 pmol) and Compound 2B (52.39 mg, 608.29 pmol) in DMF (2 mL) was added K2CO3 (126.10 mg, 912.43 pmol). Themixture was stirred at 80°C for 0.5 hr. LC-MS showed Compound 2A was consumed and 33% of desired compound was detected. The reaction mixture was filtered. The residue was purified by Prep-HPLC (column: Phenomenex Luna C18 100*30mm*5 pm; mobile phase:[H2O (0.1% TFA)-ACN]; gradient: 5%-40% B over 8.0 min) to afford CW-26 (60.05 mg, 121.93 pmol, 40.09% yield, 100% purity, TFA salt) as a white solid.

[0527] ¹H NMR (METHANOL-d4, 400 MHz): 5 ppm 8.25 (dd, J=7.94, 1.06 Hz, 1 H), 7.85 - 7.94 (m, 1 H), 7.76 - 7.84 (m, 1 H), 7.55 - 7.66 (m, 1 H), 7.42 (t, J=8.00 Hz, 1 H), 7.05 (t, 7.50 Hz, 2 H), 3.95 - 4.25 (m, 2 H), 3.32 - 3.54 (m, 2 H), 3.12 (br s, 4 H), 2.67 - 2.92 (m, 2 H), 2.36 - 2.57 (m, 2 H), 2.17 (s, 3 H), 1.19 (t, J=7.00 Hz, 3 H)

[0528] LCMS (ESI+): m / z 379.2 (M+H).CW-27s CW-27

[0529] To a solution of Compound 1 (1 g, 6.53 mmol) and EtI (1.22 g, 7.84 mmol, 626.76 pL) in DMF (40 mL) was added K2CO3 (2.71 g, 1.59 mmol) at 20°C. The mixture was stirred at 80°C for 2 hrs. LC-MS showed Compound 1 was consumed and 27% of desired compound was detected. The reaction mixture was diluted with H2O (20 mL) and extracted with EtOAc (20 mL * 3). The combined organic layers were washed with brine (50 mL * 1), dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to afford Compound 2 (1 g, crude) as a yellow oil. MS-ESI (m / z) ealed for C9HIINO3 [M+H]+: 182.1 Found 182.4. To a solution of Compound 2 (1 g, 5.52 mmol) in EtOH (20 mL) and H2O (5 mL) was added Fe (924.65 mg, 16.56 mmol) and NH4CI (1.48 g, 27.60 mmol). The mixture was stirred at 80°C for 1 hr. LC-MS showed Compound 2 wasconsumed and 76% of desired compound was detected. The reaction mixture was filtered and then the mixture was poured into saturated NaHCO solution to pH = 8 ~ 9, then the mixture was extracted with EtOAc (10 mL * 3). The combined organic layers were dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to afford Compound 3 (700 mg, crude) as a yellow oil. MS-ESI (m / z) calcd for C9H13NO [M+H]+: 152.1 Found 152.5. To a solution of Compound 3 (150 mg, 702.19 pmol) and Compound 3A (106.17 mg, 702.19 pmol) in ACN (3 mL) was added POCh (323.01 mg, 2.11 mmol, 196.36 pL). The mixture was stirred at 80°C for 12 hrs. LC-MS showed Compound 3 was consumed and 51 % of desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with EtOAc 5 mL and then was poured into saturated Na2CO3 solution to pH = 8 ~ 9, the mixture was extracted with EtOAc (5 mL * 3). The combined organic layers were dried over Na2SO4, 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 0~8% Ethyl acetate / Petroleum ether gradient @ 60 mL / min) to afford Compound 4 (120 mg, 357.67 pmol, 50.94% yield, 98% purity) as a yellow solid. MS-ESI (m / z) calcd for C18H17CIN2O2 [M+H]+: 329.1 / 331.1 Found 329.2 / 331.2. To a solution of Compound 4 (120 mg, 364.97 pmol) and Compound 4A (87.71 mg, 437.97 pmol) in DMF (2 mL) was added K2CO3 (151.32 mg, 1.09 mmol). The mixture was stirred at 80°C for 12 hrs. LC-MS showed with EtOAc (2 mL * 3). The combined organic layers were washed with brine (3 mL * 1), dried over anhydrous Na2SO4, 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 0~8% Ethyl acetate / Petroleum ether gradient @ 60 mL / min) to afford Compound 5 (120 mg, 204.63 pmol, 56.07% yield, 84% purity) as a yellow oil. MS-ESI (m / z) calcd for C28H36N4O4 [M+H]+: 493.3 Found 493.2. To a solution of Compound 5 (120 mg, 243.60 pmol) in DCM (2 mL) was added TFA (555.52 mg, 4.87 mmol, 361.90 pL). The mixture was stirred at 20°C for 1 hr. LC-MS showed Compound 5 was consumed and 82% of desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by Prep-HPLC (TFA condition; column: Phenomenex Luna C18 100*30mm*5 pm; mobile phase:[H2O (0.1% TFA) - ACN]: gradient: 10% - 40% B over 10.0 min) to afford B-27 (31.36 mg, 61.40 pmol, 88.86% yield, 99.17% purity, TFA) as a yellow gum.

[0530] 1H NMR (METHANOL-d4, 400MHz): δ ppm 8.27 - 8.18 (m, 1H), 7.93 - 7.83 (m, 1H), 7.77 (d, J = 8.1 Hz, 1H), 7.58 (t, J = 7.6 Hz, 1H), 7.27 - 7.17 (m, 1H), 7.05 (s, 1H), 6.96 (d, J = 7.9 Hz, 1H), 4.18 - 4.03 (m, 2H), 4.00 - 3.66 (m, 1H), 3.38 - 3.32 (m, 1H), 3.25 - 3.16 (m, 1H), 3.12 (br d, J = 12.4 Hz, 1H), 3.04 - 2.93 (m, 2H), 2.92 - 2.83 (m, 1H), 2.82 - 2.69 (m, 1H), 2.61 (brt, J = 11.4 Hz, 1H), 2.50 - 2.41 (m. 3H), 1.24 - 1.16 (m, 3H), 0.84 - 0.70 (m, 3H).

[0531] LCMS (ESI+): m / z 393.2 (M+H).CW-28

[0532] To a solution of Compound 1 (200 mg, 1.10 mmol) and EtI (206.59 mg, 1.32 mmol, 105.94 pL) in DMF (3 mL) was added K2CO3 (457.66 mg, 3.31 mmol). The mixture was stirred at 80°C for 1 hr. LC-MS showed Compound 1 was consumed and 33% of desired compound was detected. The reaction mixture was diluted with H2O 3 mL and extracted with EtOAc (3 mL * 3). The combined organic layers were washed with brine (5 mL * 1), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to afford Compound 2 (240 mg, crude) as a yellow oil. MS-ESI (m / z) calcd for C11H15NO3 [M+H]+: 210.1 Found 210.2. To a solution of Compound 2 (240 mg, 1.15 mmol) in EtOH (1.2 mL) and H2O (0.3 mL) was added Fe (192.16 mg, 3.44 mmol) and NH4C1 (306.77 mg, 5.74 mmol). The mixture was stirred at 80°C for 1 hr. LC-MS showed Compound 2 was consumed and 90% of desired compound was detected. The reaction mixture was filtered and the filtrate was poured into saturated NaHCCh solution to pH = 8 ~ 9 and extracted with EtOAc (3 mL * 3). The combined organic layers were dried over anhydrous Na2SO4, filteredand concentrated under reduced pressure to afford Compound 3 (180 mg, crude) as a yellow oil. MS-ESI (m / z) calcd for C11H17NO [M+H]+: 180.1 Found 180.2. To a solution of Compound 3 (100 mg, 468.13 pmol) and Compound 3A (100.70 mg, 561.75 pmol) in ACN (2 niL) was added POCI3 (215.34 mg, 1.40 mmol, 130.90 pL). The mixture was stirred at 80°C for 1 hr. LC-MS showed Compound 3 was consumed and 51 % of desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with EtOAc 3 mL and then the mixture was poured into saturated Na₂CO₃ solution to pH = 8 ~ 9, the mixture was extracted with EtOAc (3 mL * 3). The combined organic layers were dried over Na2SO4, 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 0-10% Ethyl acetate / Petroleum ether gradient @ 60 mL / min) to afford Compound 4 (90 mg, 234.56 pmol, 50.11% yield, 93% purity) as a yellow oil. MS-ESI (m / z) calcd for C20H21CIN2O2 [M+H]+: 357.1 / 359.1 Found 357.2 / 359.2. To a solution of Compound 4 (70 mg, 196.16 pmol) and Compound 4A (47.14 mg, 235.40 pmol,) in ACN (2 mL) was added K2CO3 (81.33 mg, 588.49 pmol) and Nal (5.88 mg, 39.23 pmol). The mixture was stirred at 80°C for 1 hr. LC-MS showed Compound 4 was consumed and 48% of desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with H2O 2 mL and extracted with EtOAc (2 mL * 3). The combined organic layers were washed with brine (5 mL * 1), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to afford Compound 5 (80 mg, crude) as a yellow oil. MS-ESI (m / z) calcd for C30H40N4O4 [M+H]+: 521.3 Found 521.4. To a solution of Compound 5 (80 mg, 153.65 pmol) in DCM (2 mL) was added TFA (350.39 mg, 3.07 mmol, 228.27 pL). The mixture was stirred at 20°C for 1 hr. LC-MS and HPLC showed Compound 5 was consumed and 60% of desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by Prep-HPLC (TFA condition; column: Phenomenex Luna C18 100*30mm*5 pm; mobile phase: [H2O (0.1% TFA) - ACN]; gradient: 15% - 45% B over 8.0 min) to afford CW-28 (36.93 mg, 69.08 pmol, 44.96% yield, 100% purity, TFA) as a pale yellow gum.

[0533] ¹H NMR (METHANOL-d4, 400MHz): 5 ppm 8.23 (dd, J = 1.4, 7.9 Hz, III), 7.93 - 7.85 (m, 1H), 7.78 (br d, J = 7.9 Hz, 1H), 7.64 - 7.55 (m, 1H), 7.39 (dd, J = 1.8, 8.5 Hz, 1H), 7.24 (dd, J = 2.3, 9.3 Hz, 1H), 7.18 - 7.11 (m, 1H), 4.17 - 4.01 (m, 2H), 4.01 - 3.76(m, 1H), 3.35 - 3.16 (m, 2H), 3.14 - 3.05 (m, 1H), 3.01 - 2.89 (m, 3H), 2.78 (br d,.1 = 12.6 Hz, 1H), 2.71 - 2.45 (m, 2H), 1.34 - 1.24 (m, 6H), 1.20 (dt,.1 = 2.4, 7.0 Hz, 3H), 0.78 - 0.64 (m, 3H)

[0534] LCMS (ESI+): m / z 421.3 (M+H)

[0535] To a solution of Compound 1 (1 g. 7.34 mmol, 1.01 mL) in AcOH (10 mL) was added HNO3 (810 mg, 8.36 mmol, 578.57 pL, 65% purity) at 0°C. The mixture was stirred at 20°C for 1 hr. LC-MS showed Compound 1 was consumed and 45% of desired compound was detected. The reaction mixture was poured into saturated NaHCOs solution (20 mL) and extracted with EtOAc (10 mL * 3). The combined organic layers were dried over anhydrous 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 Flash Column, Eluent of 0~3% Ethyl acetate / Petroleum ether gradient @ 80mL / min) to afford Compound 2 (330 mg, 1.78 mmol, 24.31% yield, 98% purity) as a yellow oil. MS-ESI (m / z) calcd for C9H11NO3 [M-HJ: 180.1 Found 180.5. ’ll NMR (DMSO-r / 6400MHz): 6 ppm 10.78 (br s, 1H), 7.85 (d, J = 8.6 Hz, 1H), 6.98 (d, J = 1.5 Hz, 1H), 6.89 (dd, J = 1.6, 8.6 Hz, 1H), 2.90 (td, J = 6.9, 13.8 Hz, 1H), 1.18 (d, J = 6.9 Hz, 6H) Compound 2A (450 mg, 2.31 mmol, 31.46% yield, 93% purity) as a yellow oil. MS-ESI (m / z) calcd for C9H11NO3 [M-H]: 180.0 Found 180.5Spectrum:

[0536] 1H NMR ET88248-523-P1C DMSO- 6400MHz

[0537] 5 ppm 10.66 (s, 1H), 7.80 (d, J = 8.9 Hz, 1H), 6.88 (d, J = 2.6 Hz, 1H), 6.74 (dd, J = 2.6, 8.9 Hz, 1H), 3.45 (td, J = 6.8, 13.6 Hz, 1H), 1.23 - 1.18 (m, 6H)

[0538] To a solution of Compound 2 (300 mg, 1.66 mmol) and EtI (387.36 mg, 2.48 mmol, 198.64 pL) in DMF (2 mL) was added K2CO3 (686.50 mg, 4.97 mmol). The mixture was stirred at 80°C for 12 hrs. LC-MS showed Compound 2 was consumed and 28% of desired compound was detected. The reaction mixture was diluted with H2O 3 mL and extracted with EtOAc (3 mL * 3). The combined organic layers were washed with brine (5 mL * 1), dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to afford Compound 3 (320 mg, crude) as a yellow oil. MS-ESI (m / z) calcd for C11H15NO3 [M+H]+: 210.1 Found 210.5. To a solution of Compound 3 (220 mg, 1.05 mmol) in EtOH (4 mL) and H2O (1 mL) was added Fe (176.15 mg, 3.15 mmol) and NH4CI (281.21 mg, 5.26 mmol). The mixture was stirred at 80°C for 1 hr. LC-MS showed Compound 3 was consumed and 81 % of desired compound was detected. The reaction mixture was filtered and then poured into saturated Na2CC>3 solution (5 mL) and extracted with EtOAc (3 mL * 3). The combined organic layers were dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give Compound 4 (180 mg, crude) as a yellow oil. MS-ESI (m / z) calcd for C11H17NO [M+H]+: 180.1 Found 180.2. To a solution of Compound 4 (100 mg, 468.13 ymol) and Compound 4A (100.70 mg, 561.75 ymol) in ACN (2 mL) was added POCl₃ (215.34 mg, 1.40 mmol, 130.90 pL). The mixture was stirred at 80°C for 1 hr. LC-MS showed Compound 4 was consumed and 32% of desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with EtOAc 3 mL and then poured into saturated Na2CO3 solution to pH = 8 ~ 9, the mixture was extracted with EtOAc (3 mL * 3). The combined organic layers were dried over anhydrous Na2SO4, 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 0-10% Ethyl acetate / Petroleum ether gradient @ 60 mL / min) to afford Compound 5 (70 mg, 190.28 pmol, 40.65% yield, 97% purity) as a yellow oil. MS-ESI (m / z) calcd for C20H21CIN2O2 [M+H]+: 357.1 / 359.1 Found 357.1 / 359.1. To a solution of Compound 5 (50 mg, 140.12 pmol) and Compound 5A(33.67 mg, 168.14 pmol) in ACN (2 mL) was added K2CO3 (58.10 mg, 420.35 pmol) and Nal (4.20 mg, 28.02 pmol). The mixture was stirred at 80°C for 1 hr. LC-MS showed Compound 5 was consumed and 49% of desired compound was detected. The reaction mixture was diluted with H2O (1 mL) and extracted with EtOAc (2 mL * 3). The combined organic layers were washed with brine (2 mL * 1), dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to afford Compound 6 (70 mg, crude) as a yellow oil. MS-ESI (m / z) calcd for C30H40N4O4 [M+H]+: 521.3 Found 521.4. To a solution of Compound 6 (70 mg, 134.44 pmol) in DCM (2 mL) was added TFA (306.60 mg, 2.69 mmol, 199.74 pL). The mixture was stirred at 20°C for 1 hr. LC-MS and HPLC showed Compound 6 was consumed and 67% of desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by Prep-HPLC (TFA condition; column: Phenomenex Luna C18 100*30mm*5 μm; mobile phase: [H2O (0.1% TFA) - ACN]; gradient: 15% - 45% B over 8.0 min) to afford CW-29 (21.06 mg, 39.23 pmol, 29.18% yield, 99.58% purity, TFA) as a pale yellow gum. Spectrum:

[0539] ¹H NMR (METHANOL-d4, 400MHz)

[0540] 5 ppm 8.23 (d, J = 8.0 Hz, 1H), 7.93 - 7.84 (m, 1H), 7.77 (d, J = 8.1 Hz, 1H), 7.63 - 7.55 (m, 1H), 7.31 - 7.22 (m, 1H), 7.08 (s, 1H), 7.03 (br d, J = 8.0 Hz, 1H), 4.21 - 4.03 (m, 2H), 3.97 - 3.74 (m, 1H), 3.38 - 3.15 (m, 2H), 3.10 (br d,.1 = 11.6 Hz, 1H), 3.04 - 2.92 (m, 2H), 2.88 - 2.76 (m, 1H), 2.76 - 2.59 (m, 2H), 2.58 - 2.47 (m, 1H), 1.32 (d, J = 6.9 Hz, 6H), 1.21 (dt, J = 2.9, 7.0 Hz, 3H). 0.76 - 0.69 (m. 3H)

[0541] LCMS (ESI+): m / z 421.3 (M+H).CW-30L

[0542] To a solution of Compound 1 (1 g, 4.59 mmol) and EtI (858.50 mg, 5.50 mmol, 440.26 pL) in DMF (20 mL) was added K2CO3 (1.90 g, 13.76 mmol). The mixture was stirred at 80°C for 12 hrs. TLC showed Compound 1 was consumed and one major new spot with lower polarity was detected. The reaction mixture was diluted with H2O (20 mL) and extracted with EtOAc (20 mL * 3). The combined organic layers were washed with brine (50 mL * 1), dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to afford Compound 2 (1.1 g, crude) as a yellow oil. 1H NMR (DMSO-t / 6400MHz): 6 ppm 7.52 - 7.44 (m, 1H), 7.40 - 7.34 (m, 2H), 4.21 (q, J = 7.0 Hz, 2H), 1.28 (t,.1 = 7.0 Hz, 3H). To a solution of Compound 2 (1.1 g, 4.47 mmol) and Compound 2A (901.47 mg, 5.36 mmol) in dioxane (20 mL) and H2O (4 mL) was added Pd(dppf)Ci2. CH2Ci2 (365.08 mg, 447.05 pmol) and K2CO3 (1.85 g, 13.41 mmol). The mixture was stirred at 80°C for 12 hrs under N2. LC-MS showed Compound 2 was consumed and 47% of desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0~5% Ethyl acetate / Petroleum ether gradient @ 100 mL / min) to afford Compound 3 (900 mg, 4.34 mmol, 97.15% yield) as a yellow oil. MS-ESI (m / z) calcd for C11H13NO3 [M+H]+: 208.1 Found 208.4. 1H NMR (DMSO-J6400MHz): 5 ppm 7.49 (t, J = 8.1 Hz, 1H), 7.24 (d, J = 8.4 Hz, 1H), 7.01 (d, J = 7.8 Hz, 1H), 5.21 (s, 1H), 4.91(s, 1H), 4.17 (q, J = 7.0 Hz, 2H), 2.01 (s, 3H), 1.28 (t, J = 6.9 Hz, 3H). To a solution of Compound 3 (500 mg, 2.41 mmol) in MeOH (10 mL) was added Pd / C (300 mg, 10% purity) at 25°C. The mixture was stirred at 25°C for 4 hrs under H2 at 15psi. LC-MS showed Compound 3 was consumed and 84% of desired compound was detected. The reaction mixture was filtered and then dried under vacuum to afford Compound 4 (350 mg, crude) as a yellow oil. MS-ESI (m / z) calcd for C11H17NO [M+H]+: 180.1 Found 180.4. To a solution of Compound 4 (100 mg, 468.13 pmol) and Compound 4A (100.70 mg, 561.75 pmol) in ACN (3 mL) was added POC13 (215.34 mg, 1.40 mmol, 130.90 pL). The mixture was stirred at 80°C for 12 hrs. LC-MS showed Compound 4 was consumed and 55% of desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was diluted with EtOAc (3 mL) and then poured into saturated Na2CO3 solution to pH = 8 ~ 9, the mixture was extracted with EtOAc (3 mL * 3). The combined organic layers were dried over Na2SO4, 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 0~5% Ethyl acetate / Petroleum ether gradient @ 60 mL / min) to afford Compound 5 (90 mg, 237.08 pmol, 50.64% yield, 94% purity) as a yellow solid. MS-ESI (m / z) calcd for C20H21CIN2O2 [M+H]+: 357.1 / 359.1 Found 357.2 / 359.2. To a solution of Compound 5 (80 mg, 224.19 pmol) and Compound 5 A (44.90 mg, 224.19 pmol) in ACN (2 mL) was added K2CO3 (92.95 mg, 672.56 pmol) and Nal (3.36 mg, 22.42 pmol). The mixture was stirred at 80°C for 12 hrs. LC-MS showed Compound 5 was consumed and 48% of desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by flash silica gel chromatography (ISCO®; 4 g SepaFlash® Silica Flash Column, Eluent of 0-10% Ethyl acetate / Petroleum ether gradient @ 60 mL / min) to afford Compound 6 (60 mg, 102.56 pmol, 45.75% yield, 89% purity) as a yellow oil. MS-ESI (m / z) calcd for C30H40N4O4 [M+H]+: 521.3 Found 521.4. To a solution of Compound 6 (60 mg, 115.24 pmol) in DCM (1 mL) was added TFA (262.80 mg, 2.30 mmol, 171.20 pL). The mixture was stirred at 20°C for 1 hr. LC-MS and HPLC showed Compound 6 was consumed and 88% of desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by Prep-HPLC (TFA condition; column: Phenomenex Luna C18 100*30mm*5um; mobile phase: [H2O (0.1%TFA) - ACN]; gradient: 15% - 45% B over 8.0 min) to afford CW-30 (26.06 mg, 48.75 pmol, 42.30% yield, 100% purity, TFA) as a white solid.

[0543] 1H NMR (METHANOL-d4 400MHz): 6 ppm 8.23 (br d, J = 7.9 Hz, 1H), 7.96 - 7.85 (m, 1H), 7.80 (brd, J = 8.1 Hz, 1H), 7.64 - 7.55 (m, 1H), 7.51 (t, J = 8.1 Hz, 1H), 7.19 - 7.10 (m, 1H), 7.08 - 7.01 (m, 1H), 4.17 - 3.97 (m, 2H), 3.79 - 3.60 (m, 1H), 3.36 - 3.21 (m, 2H), 3.19 - 3.10 (m, 1H), 3.08 - 2.88 (m, 3H), 2.85 - 2.51 (m, 3H), 1.33 - 1.26 (m, 3H), 1.23 - 1.07 (m, 6H), 0.90 - 0.65 (m, 3H)

[0544] LCMS (ESI+): m / z 421.3 (M+H)

[0545] To a solution of Compound 1 (6 g, 26.05 mmol) and TEA (5.52 g, 54.58 mmol, 7.60 mL, 2.09 eq) in DCM (60 mL) was added Alloc-Cl (3.45 g, 28.66 mmol, 3.04 mL) at 0°C, then the mixture was stirred at 0°C for 1 hr. LCMS showed 9% of Compound 1 wasremained and 48% of desired product was detected. The reaction was concentrated under vacuum. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0-30% Ethyl acetate / Petroleum ether gradient @ 100 mL / min) to afford Compound 2 (4.69 g, 14.92 mmol, 57.26% yield) as a colorless oil. MS-ESI (m / z) calcd for C15H26N2O5 [M+H]+: 315.2 Found 315.3. 1H NMR (CHLOROFORM-T / 6400MHZ): 5 ppm 5.87 - 6.02 (m, 1 H), 5.21 - 5.37 (m, 2 H), 4.63 (br s, 2 H), 4.36 (br s, 1 H),3.84 -4.09 (m, 3 H), 3.59 - 3.73 (m, 1 H), 3.40 (br s, 1 H), 3.11 (br s, 1 H), 2.99 (td, J=12.76, 3.38 Hz, 1 H), 2.85 (br d, J=1.38 Hz, 1 H), 1.79 - 1.93 (m, 1 H), 1.73 (br d, J=2.00 Hz, 1 H), 1.47 (s, 9 H). To a solution of Compound 2 (3 g, 9.54 mmol) in DCM (30 mL) was added CBr4 (4.11 g, 12.41 mmol) and PPh3 (3.25 g, 12.41 mmol) at 0°C, then the mixture was stirred at 0°C for 1 hr. TLC showed Compound 2 was consumed and desired product was detected. The reaction was concentrated under vacuum. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0-20% Ethyl acetate / Petroleum ether gradient @ 40 mL / min) to afford Compound 3 (2.9 g, 6.23 mmol, 65.25% yield, 81% purity) as a yellow oil. MS-ESI (m / z) calcd for C15H25BrN2O4 [M+H-100]+: 277.1 / 279.1 Found 277.2 / 279.2. To a solution of Compound 3 (2.9 g, 7.69 mmol) in NaOH (6 M, 55.77 mL) was added Compound 3A (1.35 g, 7.69 mmol) and TBAB (247.80 mg, 768.67 pmol) at 20°C and the reaction was stirred at 20°C for 12 hrs. LCMS showed Compound 3 was consumed and desired product was detected. The reaction was quenched with H2O (50 mL), extracted with EtOAc (50 mL * 3), the organic layer was dried over anhydrous Na2SC>4, filtered and the filtrate was concentrated under vacuum. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0-20-30% Ethyl acetate / Petroleum ether gradient @ 100 mL / min) to afford Compound 4 (1.56 g, 3.31 mmol, 43.04% yield, 100% purity) as a colorless oil. MS-ESI (m / z) calcd for C21H37N5O7 [M+H]+: 472.3 Found 472.3. To a solution of Compound 4 (160 mg, 339.31 pmol) in THF (1.5 mL) and H2O (1.5 mL) was added PPh3 (266.99 mg, 1.02 mmol) at 20°C, then the mixture was stirred at 70°C for 2 hrs. LCMS showed Compound 4 consumed and 41 % of desired product was detected. 2 mL 1 M HO was added into the reaction, then stirred at 20°C for 1 hr, then extracted with EtOAc (3 mL * 2), the organic layer was discarded, the aqueous layer was basified with saturated Na2CO3 solution to pH = 8, then extracted with EtOAc (3 mL * 3), the organic layer was dried over anhydrous Na2SO4, filtered and the filtrate was concentrated under vacuum to affordCompound 5 (115 mg, crude) as a colorless oil and it was used directly without further purification. MS-ESI (m / z) calcd for C21H39N3O7 [M+H]+: 446.3 Found 446.2. To a solution of Compound 5 (115 mg, 258.11 pmol) and Compound 5A (103.47 mg, 258.11 pmol) in DMF (2 niL) was added HATU (98.14 mg, 258.11 pmol) and DIEA (66.72 mg, 516.22 pmol, 89.92 pL) at 20°C, then the mixture was stirred at 20°C for 0.5 hr. LCMS showed Compound 5 was consumed and 12% of desired product was detected. The reaction was filtered. The filtrate was purified by Prep-HPLC (TEA condition, column: Phenomenex Luna C18 100*30mm*5 pm; mobile phase: [H2O (0.1% TEA) - ACN]; gradient: 35% - 65% B over 8.0 min) to afford Compound 6 (105 mg, 119.87 pmol, 46.44% yield, 94.57% purity) as a pale yellow solid. MS-ESI (m / z) calcd for C40H54ClN7O8S [M+H]+: 828.3 Found 828.2. To a solution of Compound 6 (95 mg, 114.68 pmol) in DCM (2 mL) was added DMBA (44.76 mg, 286.69 pmol) and Pd (PPh3)4 (13.25 mg, 11.47 pmol) at 20°C, then the mixture was stirred at 20°C for 12 hrs under N2. LCMS showed Compound 6 was consumed and 17% of desired product was detected. The reaction was concentrated under vacuum. The residue was purified by Prep-HPLC (TFA condition, column: Phenomenex Luna C18 100*30mm*5 pm; mobile phase: [H2O (0.1% TFA) - ACN]; gradient: 15% - 65% B over 8.0 min) to afford Compound 7 (85 mg, 99.03 pmol, 86.35% yield, TFA) as a yellow liquid. MS-ESI (m / z) calcd for C36H50CIN7O6S [M+H]+:744.3 / 745.3 / 746.3 Found 744.3 / 745.3 / 746.2. To a solution of Compound 7 (65 mg, 75.73 pmol, TFA) and Compound 7A (29.88 mg, 90.87 pmol) in ACN (1 mL) was added K2CO3 (31.40 mg, 227.18 pmol) and Nal (1.14 mg, 7.57 pmol) at 20°C, then the mixture was stirred at 80°C for 12 hrs. LCMS showed 35% of Compound 7 was remained and 26% of desired product was detected. The reaction mixture was filtered. The filtrate was purified by Prep-HPLC (column: Phenomenex Luna C18 100*40mm*5 pm; mobile phase: [H2O (0.1% TFA) - ACN]; gradient: 30% - 70% B over 8.0 min) to afford Compound 8 (50 mg) as a yellow liquid. MS-ESI (m / z) calcd for C54H66CIN9O8S [M+H]+: 1036.4 Found 1036.4. To a solution of Compound 8 (50 mg, 48.23 pmol) in DCM (1 mL) was added TFA (307.00 mg, 2.69 mmol, 0.2 mL) at 20°C, then the mixture was stirred at 20°C for 1 hr. LCMS showed Compound 8 was consumed and 54% of desired product was detected. The reaction mixture was concentrated under vacuum. The residue was purified by Prep-HPLC (column: Phenomenex Luna C18 100*40mm*5 pm; mobile phase: [H2O (0.1% TFA) - ACN]; gradient: 30% - 50% B over 10.0 min) to afford CW-31 (12.15 mg, 11.41 pmol, 23.65% yield, 98.64% purity, TFA) as a colorless gum.

[0546] 1H NMR (CHLOROFORM-c / 6400MHz): 5 ppm 8.28 (br d, J=7.88 Hz, 1 H) 8.05 - 8.18 (m, 1 H) 7.72 - 7.89 (m, 2 H) 7.49 - 7.56 (m, 1 H) 7.43 (br dd, J=7.88, 3.25 Hz, 2 H) 7.32 - 7.37 (m, 2 H) 7.02 - 7.12 (m, 1 H) 6.84 - 6.95 (m, 2 H) 4.60 - 4.90 (m, 1 H) 3.95 -4.19 (m, 2 H) 3.68 - 3.92 (m, 2 H) 3.52 - 3.67 (m, 12 H) 3.46 - 3.52 (m, 3 H) 3.17 - 3.45 (m, 8 H) 2.57 - 2.78 (m, 3 H) 2.37 - 2.48 (m, 6 H) 1.73 - 1.94 (m, 2 H) 1.70 (s, 3 H) 1.21 (t,.1=6.88 Hz, 3 H)

[0547] LCMS (ESI+): m / z 936.3 (M+H)CW-32

[0548] To a solution of Compound 5 (115 mg, 258.11 pmol) and Compound 5A (103.47 mg, 258.11 pmol) in DMF (2 mL) was added HATU (98.14 mg, 258.11 pmol) and DIEA (66.72 mg, 516.22 pmol, 89.92 pL) at 20°C, then the mixture was stirred at 20°C for 0.5 hr. LCMS showed Compound 5 was consumed and 12% of desired product was detected. The reaction was filtered. The filtrate was purified by Prep-HPLC (TFA condition, column: Phenomenex Luna C18 100*30mm*5 pm; mobile phase: [H2O (0.1% TFA) - ACN]; gradient: 35% - 65% B over 8.0 min) to afford Compound 6 (105 mg, 119.87 pmol, 46.44% yield, 94.57% purity) as a pale yellow solid. MS-ESI (m / z) calcd for C40H54CIN7O8S [M+H]+: 828.3 Found 828.2. To a solution of Compound 6 (20 mg, 24.14 pmol) in DCM(1 mL) was added TFA (307.00 mg, 2.69 mmol) at 20°C, then the mixture was stirred at 20°C for 1 hr. LCMS showed Compound 6 was consumed and 75% of desired product was detected. The reaction was concentrated under vacuum to afford Compound 7 (20 mg, crude, TFA) as a yellow liquid and it was used directly without further purification. MS-ESI (m / z) calcd for C35H46CIN7O6S [M+H]+:728.3 / 729.3 / 730.3 Found 728.2 / 729.2 / 730.2. To a solution of Compound 7 (20 mg, 23.74 pmol, TFA) and Compound 7A (1.61 mg, 47.49 pmol) in ACN (1 mL) was added K2CO3 (9.84 mg, 71.23 pmol) at 20°C, then the mixture was stirred at 80°C for 12 hrs. LC-MS showed Compound 7 was consumed and 39% of desired compound was detected. The reaction mixture was diluted with H2O (3 mL) and extracted with EtOAc (3 mL *3), dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to afford Compound 8 (20 mg, crude) as a yellow oil. MS-ESI (m / z) calcd for C53H62CIN9O8S [M+H]+:1020.4 / 1021.4 Found 1020.4 / 1021.4. To a solution of Compound 8 (20 mg, 19.60 pmol) in DCM (1 mL) was added DMBA (7.65 mg, 48.99 pmol) and Pd (PPh3)4 (2.26 mg, 1.96 pmol). The mixture was stirred at 20°C for 2 hrs under N2. LC-MS showed Compound 8 was consumed and 30% of desired compound was detected. The reaction mixture was concentrated under vacuum. The residue was purified by Prep-HPLC (column: Phenomenex Luna C18 100*30mm*5 pm; mobile phase: [H2O (0.1% TFA) - ACN]; gradient: 20% - 60% B over 8.0 min) to afford CW-32 (11.27 mg, 10.56 pmol, 53.88% yield, 98.43% purity, TFA) as a colorless gum.

[0549] 1H NMR (CHLOROFORM-d6 400MHz): 8 ppm 8.24 - 8.32 (m, 1 H), 7.80 -7.88 (m, 1 II), 7.71 - 7.79 (m, 2 II), 7.46 - 7.56 (m, 1 II), 7.30 - 7.45 (m, 4 II), 6.96 - 7.13 (m, 1 H), 6.77 - 6.94 (m, 2 H), 4.71 (t, J=7.00 Hz, 1 H), 3.94 - 4.16 (m, 2 H), 3.81 - 3.94 (m, 1 H), 3.70 - 3.78 (m, 2 H), 3.49 - 3.69 (m, 16 H), 3.34 - 3.49 (m, 3 H), 3.13 - 3.33 (m, 3 H), 2.65 (d, 1=9.38 Hz, 3 H), 2.33 - 2.48 (m, 6 H), 2.07 - 2.31 (m, 1 H), 1.74 - 1.87 (m, 1 H), 1.69 (s, 3 H), 1.11 - 1.28 (m, 3 H)

[0550] LCMS (ESI+): m / z 936.3 (M+H)CW-33

[0551] To a solution of Compound 4 (150 mg, 456.22 pmol) and Compound 4A (78.59 mg, 912.43 pmol) in DMF (2 mL) was added K2CO3 (189.15 mg, 1.37 mmol). The mixture was stirred at 80°C for 12 hrs. LC-MS showed Compound 4 was consumed and 37% of desired compound was detected. The reaction mixture was diluted with H2O (5 mL) and extracted with EtOAc (5 mL * 3). The combined organic layers were washed with brine (5 mL * 2), dried over anhydrous Na2SO4, 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 10% Dichloromethane: Methanol gradient @ 40 mL / min) to afford Compound 5 (140 mg, 369.91 pmol, 81.08% yield) as a yellow solid. MS- ESI (m / z) calcd for C22H26N4O2 [M+H]+: 379.2 Found 379.2. To a solution ofCompound 5 (110 mg, 290.65 pmol) and Compound 5A (108.89 mg, 348.78 pmol) in ACN (2 mL) was added K2CO3 (120.51 mg, 871.94 pmol). The mixture was stirred at 80°C for 2 hrs. LC-MS showed Compound 5 was consumed and 87% of desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove solvent. The reaction mixture was diluted with H2O (5 mL) and extracted with EtOAc (5 mL * 3), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to afford Compound. 6 (170 mg, crude) as a yellow solid. MS-ESI (m / z) calcd for C33H47N5O6 [M+H]+: 610.3 Found 610.4. To a solution of Compound 6 (170 mg, 278.80 pmol) in DCM (1 mL) was added TFA (1.30 g, 11.44 mmol, 850.00 pL). The mixture was stirred at 20°C for 12 hrs. LC-MS showed Compound 6 was consumed and 88% of desired compound was detected. The reaction mixture was concentrated under vacuum. The residue was purified by Prep-HPLC (column: Phenomenex Luna C18 100* 30mm* 5 pm; mobile phase: [H2O (0.1% TFA) - ACN]; gradient: 10% - 45% B over 8.0 min) to afford Compound 7 (110 mg, 176.38 pmol, 63.26% yield, TFA) as a colorless solid. MS-ESI (m / z) calcd for C28H39N5O4 [M+H]+: 510.3 Found 510.3. To a solution of Compound 7 (100 mg, 196.22 pmol) and Compound 7A (94.39 mg, 235.46 pmol) in DMF (1 mL) was added HATU (89.53 mg, 235.46 pmol) and DIEA (76.08 mg, 588.65 pmol, 102.53 pL). The mixture was stirred at 20°C for 1 hr. LC-MS showed Compound 7 was consumed and 25% of desired compound was detected. The reaction mixture was filtered. The filtrate was purified by Prep-HPLC (column: Phenomenex Luna C18 100*30mm*5 pm; mobile phase: [H2O (0.1% TFA) - ACN]; gradient: 15% - 65% B over 8.0 min) to afford C-33 (70.54 mg, 69.77 pmol, 35.56% yield, 99.55% purity, TFA) as a pale yellow solid.

[0552] 1H NMR (METHANOL-d4 400MHz): 5 ppm 8.16 - 8.24 (m, 1 H) 7.81 - 7.90 (m, 1 H) 7.74 (d, J=8.13 Hz, 1 H) 7.53 - 7.60 (m, 1 H) 7.37 - 7.48 (m, 4 H) 7.18 - 7.29 (m, 1 H) 6.99 - 7.08 (m, 1 H) 6.88 - 6.97 (m, 1 H) 4.58 - 4.67 (m, 1 H) 3.97 - 4.15 (m, 2 H) 3.79 -3.85 (m, 2 H) 3.68 (s, 4 H) 3.56 - 3.63 (m, 2 H) 3.32 - 3.52 (m, 10 H) 3.27 (br d, J=1.00 Hz, 2 H) 2.72 - 2.86 (m, 2 H) 2.68 - 2.71 (m, 3 H) 2.57 - 2.65 (m, 2 H) 2.40 - 2.46 (m, 6 H) 1.69 (d, J=2.38 Hz, 3 H) 1.17 (t, J=6.94 Hz, 3 H)

[0553] LCMS (ESI+): m / z 892.2 (M+H)

[0554] To a solution of Compound 1 (10 g, 59.12 mmol) and K2CO3 (24.51 g, 177.37 mmol) in DMF (100 mL) was added EtI (10.14 g, 65.04 mmol, 5.20 mL) at 20°C, then the reaction was stirred at 80°C for 1 hr. LC-MS showed Compound 1 was consumed and 32% of desired product was detected. The reaction was quenched with H2O (100 mL), extracted with EtOAc (100 mL * 3), the organic layer was dried over anhydrous Na2SC>4, filtered and the filtrate was concentrated under vacuum. The residue was purified by flash silica gel chromatography (ISCO®; 12 g SepaFlash® Silica Flash Column, Eluent of 0-50% Ethyl acetate / Petroleum ether gradient @ 40 mL / min) to afford Compound 2 (11.6 g, 58.83 mmol, 99.50% yield) as a yellow solid. MS-ESI (m / z) calcd for C9H11NO4 [M-H]: 196.1 Found 196.1. 1H NMR (CHLOROFORM-d 400MHz): 6 ppm 7.82 (d, 7=2.00 Hz, 1 H), 7.51 (dd, 7=8.57, 2.06 Hz, 1 H), 7.05 (d, 7=8.50 Hz, 1 H), 4.67 (s, 2 H), 4.18 (q, 7=7.00 Hz, 2 H), 1.47 (t, 7=7.00 Hz, 3 H). To a solution of Compound 2 (1 g, 5.07 mmol) in Tol. (5 mL) was added Compound 2A (1.21 g, 5.07 mmol) and NaOH (6 M, 10 mL), TBAB (163.48 mg, 507.13 pmol) at 20°C and the reaction was stirred at 20°C for 12 hrs. LC-MS showed Compound 2 was consumed and 61% of desired compound was detected. The reaction wasquenched with H2O (10 mL), extracted with EtOAc (10 mL * 3), the organic layer was dried over anhydrous Na2SO4, filtered and the filtrate was concentrated under vacuum. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash ® Silica Flash Column, Eluent of 30% Ethyl acetate / Petroleum ether gradient @ 80 mL / min) to afford Compound 3 (1 g, 2.82 mmol, 55.65% yield) as a colorless oil.

[0555] 1H NMR (METHANOL-d4 400MHz): 8 ppm 7.75 - 7.80 (m, 1 H), 7.53 - 7.59 (m, 1 H), 7.23 (d, J=8.63 Hz, 1 H), 4.54 (s, 2 H), 4.17 - 4.23 (m, 2 H), 3.52 - 3.85 (m, 10 H), 3.36 (t, J=4.88 Hz, 2 H), 1.42 (t, J=6.94 Hz, 3 H). To a solution of Compound 3 (500 mg, 1.41 mmol) in EtOH (10 mL) and H2O (2 mL) was added Fe (236.39 mg, 4.23 mmol) and NH4Cl (226.43 mg, 4.23 mmol). The mixture was stirred at 80°C for 0.5 hr. LC-MS showed Compound 3 was consumed and 74% of desired compound was detected. The reaction mixture was filtered and diluted with H2O (20 mL) and extracted with EtOAc (20 mL * 3). The combined organic layers were washed with brine (20 mL * 2), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 10 g SepaFlash® Silica Flash Column, Eluent of 30% Ethyl acetate / Petroleum ether gradient @ 80 mL / min) to afford Compound 4 (650 mg, 2.00 mmol, 71.01% yield) as a yellow oil. MS-ESI (m / z) ealed for C15H24N4O4 [M+H]+: 325.2 Found 325.4. To a solution of Compound 4 (217.32 mg, 1.02 mmol) in ACN (6 mL) was added DIEA (143.44 mg, 1.11 mmol, 193.31 pL). Then PC13 (152.42 mg, 1.11 mmol) was added at 20°C. The mixture was stirred at 20°C for 0.1 hr. Then Compound 4A (300 mg, 924.86 pmol) in ACN (6 mL) was added. The mixture was stirred at 60°C for 12 hrs. LC-MS showed Compound 4 was consumed and 48% of desired compound was detected. The reaction mixture was diluted with NaHCO3 (5 mL) and extracted with EtOAc (3 mL * 3). The combined organic layers were washed with brine (10 mL * 2), dried over anhydrous 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 Flash Column, Eluent of 50% Ethyl acetate / Petroleum ether gradient @ 60 mL / min) to afford Compound 5 (260 mg, 517.97 pmol, 56.01% yield) as a pale yellow solid. MS-ESI (m / z) ealed for C24H28CIN5O5 [M+H]+: 502.2 / 504.2 Found 502.1 / 504.1. To a solution of Compound 5 (140 mg, 278.91 pmol) and Compound 5A (55.86 mg, 278.91 pmol) in ACN (3 mL) was added K2CO3 (115.64 mg, 836.72 pmol). The mixture was stirred at 80°C for 12 hrs. LC-MS showed Compound 5 was consumed and 87% of desired compound wasdetected. The reaction mixture was diluted with H2O (10 mL) and extracted with EtOAc (10 mL * 3), dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 10 g SepaFlash ® Silica Flash Column, Eluent of 30% Ethyl acetate / Petroleum ether gradient @ 60 mL / min) to afford Compound 6 (180 mg, 270.36 pmol, 96.94% yield) as a yellow oil. MS-ESI (m / z) calcd for C34H47N7O7 [M+H]+: 666.4 Found 666.5. To a solution of Compound 6 (120 mg, 180.24 pmol) in THF (2 mL) and H2O (2 mL) was added PPh3 (141.82 mg, 540.72 pmol). The mixture was stirred at 80°C for 2 hrs. LC-MS showed Compound 6 was consumed and 35% of desired compound was detected. 2 mL 2M HC1 was added into the reaction, then stirred at 20°C for 1 hr, then extracted with EtOAc (5 mL * 2), the organic layer was discarded, the aqueous layer was basified with saturated Na2CO3 solution to pH = 8, then extracted with EtOAc (5 mL * 3), the organic layer was dried over anhydrous Na2SO4, filtered and the filtrate was concentrated under vacuum to afford Compound 7 (80 mg, 125.04 pmol, 69.38% yield) as a yellow oil. MS-ESI (m / z) calcd for C34H49N5O7 [M+H]+: 640.4 Found 640.3. To a solution of Compound 7 (80 mg, 125.04 pmol) and Compound 7A (50.13 mg, 125.04 pmol) in DMF (2 mL) was added HATU (57.05 mg, 150.05 pmol) and DIEA (48.48 mg, 375.13 pmol, 65.34 pL). The mixture was stirred at 20°C for 0.5 hr. LC-MS showed Compound 7 was consumed and 25% of desired compound was detected. The reaction mixture was diluted with H2O (5 mL) and extracted with EtOAc (3 mL * 3). The combined organic layers were washed with brine (5 mL * 2), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to afford Compound 8 (120 mg, crude) as a yellow oil. MS-ESI (m / z) calcd for C53H64C1N9O8S [M+H]+: 6.73 mmol, 0.5 mL). The mixture was stirred at 20°C for 1 hr. LC-MS showed Compound 8 was consumed and...

Claims

Having described the invention, we claim:

1. A compound of formula (I):R4or a pharmaceutically acceptable salt, tautomer, or solvate thereof; whereinR1is absent, halogen, alkyl, haloalkyl, or alkoxy;R2aand R2bare each independently -H, halogen, alkyl, or haloalkyl, or alternatively R2aand R2btogether form a cycloalkyl or heterocyclyl;R3and R5are each independently -H, halogen, alkyl, haloalkyl, or oxo;R4is absent, -H, halogen, alkyl, haloalkyl, or -C(O)-alkyl optionally substituted with one or more halogen;R6and R7are each independently -H, halogen, alkyl, or haloalkyl;R8is absent, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, -alkylene-OH optionally substituted with one or more halogen, -CN, -C(O)-N(R10)2, -C(O)O-alkyl optionally substituted with one or more halogen, or carboxyl;R9is -H, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, or -O-alkylene-alkynyl optionally substituted with one or more halogen;each R10is independently -H, alkyl, or haloalkyl;X is N or O; oralternatively, one of R1, R2a, R2b, R3, R4, R5, R6, R8, or R9is substituted with a linker that is optionally linked to a target protein ligand; andwherein at least one of R1, R2a, R2b, R3, R4, R5, R6, R7, or R8is not absent or -H if X is N and R9is alkoxy, such as ethoxy.

2. The compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof of claim 1, wherein R1is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, or Ci-Ce alkoxy.

3. The compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof of claim 1 or claim 2, wherein R1is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, or ethoxy.

4. The compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 1 to 3, wherein R2aand R2bare each independently -H, halogen Ci-Ce alkyl, or Ci-Ce haloalkyl, or R2aand R2btogether form a C3-C6 cycloalkyl.

5. The compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 1 to 4, wherein R2a-H, -F, -CF3, methyl, ethyl, or isopropyl and R2bis -H, or R2aand R2btogether form cyclopropyl.

6. The compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 1 to 5, wherein R3is -H, halogen, Ci-C8 alkyl, Ci-Ce haloalkyl, or oxo.

7. The compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 1 to 6, wherein R3is -H, -F, -CF3, methyl, ethyl, isopropyl, or oxo.

8. The compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 1 to 7, wherein X is N and R4is -H, Ci-Ce alkyl, or -C(O)-(Ci-Ce alkyl).

9. The compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 1 to 8, wherein X is N and R4is -H, methyl, ethyl, isopropyl, -C(O)methyl, -C(O)ethyl, or -C(O)isopropyl.

10. The compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 1 to 7, wherein X is O and R4is absent.

11. The compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 1 to 10, wherein R5is -H, halogen, Ci-Ce alkyl, or Ci-Ce haloalkyl.

12. The compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 1 to 11, wherein R5is -H, -F, -CF3, methyl, ethyl, or isopropyl.

13. The compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 1 to 12, wherein R6is H, halogen, Ci-Ce alkyl, or Ci-Ce haloalkyl.

14. The compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 1 to 13, wherein R6is -H, -F, -CF3, methyl, ethyl, or isopropyl.

15. The compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 1 to 14, wherein R7is -H, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl.

16. The compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 1 to 15, wherein R7is -H, -F, -CF3, methyl, ethyl, or isopropyl.

17. The compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 1 to 16, wherein R8is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, Ci-C6alkoxy, -(Ci-C6alkylene)-OII, -CN, -C(O)-N(H)(Ci-C6alkyl), -C(O)O-(Ci-Ce alkyl), or carboxyl.

18. The compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 1 to 17, wherein R8is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, ethoxy, propoxy, -methylene-OH, ethylene-OH, -propylene- OH, -CN, -C(O)-N(H)(methyl), -C(O)-N(H)(ethyl), -C(O)-N(H)(propyl), -C(O)O-(methyl), -C(O)O-(ethyl), -C(O)O-(propyl), or carboxyl.

19. The compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 1 to 18, wherein R9is -H, halogen, Ci-Ce alkyl, Ci-C& alkoxy, or -O-(Ci-Ce alkylene)-alkynyl.

20. The compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 1 to 1, wherein R9is -H, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, ethoxy, propoxy, or -O-(methylene)-ethynyl.

21. The compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof of claim 1, whereinR1is absent, halogen, alkyl, haloalkyl, or alkoxy;R2ais alkyl or haloalkyl;R2bis -H;R3, R5, R6, and R7are each independently -H, halogen, alkyl, or haloalkyl; R4is -H;R8is absent, halogen, alkyl, haloalkyl, -alkylene-OH, -CN, -C(O)-N(R10)2, or -C(O)O- alkyl;R9is alkoxy or -O-alkylene-alkynyl;each R10is independently -H or alkyl; andXis N; oralternatively, one of R1, R2a, R2b, R3, R4, R5, R6, R8, or R9is substituted with a linker that is optionally linked to a target protein ligand.

22. The compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof of claim 1, whereinR1is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, Ci-Ce alkoxy; R2ais Ci-C6alkyl or Ci-C6haloalkyl;R2bis -H;R3, R5, R6, and R7are each independently -H, halogen, C1-C5 alkyl, or Ci-Ce haloalkyl;R4is -H;R8is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, Ci-Ce alkoxy, -(Ci-Ce alkylene)-OH, -CN, -C(0)-N(H)(CI-C6 alkyl), -C(O)O-(Ci-C6 alkyl), or carboxyl; and R9is Ci-Ce alkoxy or -O-(Ci-C6alkylene)-alkynyl; andX is N; oralternatively, one of R1, R2a, R2b, R3, R4, R5, R6, R8, or R9is substituted with a linker that is optionally linked to a target protein ligand.

23. The compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof of claim 1, whereinR1is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, or ethoxy;R2ais methyl, ethyl, isopropyl, or -CF3;R2bis -H;R3, R5, R6, and R7are each independently -H, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, or -CF3;R4is -H;R8is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, ethoxy, propoxy, -methylene-OH, ethylene-OH, -propylene-OH, -CN, -C(O)-N(H)(methyl), -C(O)-N(H)(ethyl), -C(O)-N(H)(propyl), -C(O)O-(methyl), -C(O)O-(ethyl), -C(O)O-(propyl), or carboxyl;R9is methoxy, ethoxy, propoxy, or -O-(methylene)-ethynyl; and X is N; oralternatively, one of R1, R2a, R2b, R3, R4, R5, R6, R8, or R9is substituted with a linker that is optionally linked to a target protein ligand.

24. The compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 1 to 23, comprising an atropisomer of the compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof, wherein the atropisomer is substantially free of its opposite enantiomer or pharmaceutically acceptable salt, tautomer, or solvate thereof.

25. The compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof of claim 24, wherein the atropisomer is a P isomer.

26. A compound of formula (IA):(IA) or a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinR1is absent, halogen, alkyl, haloalkyl, or alkoxy;R2ais -H, halogen, alkyl, or haloalkyl;R3is -H, halogen, alkyl, or haloalkyl;R4is -H, halogen, alkyl, haloalkyl, or -C(O)-alkyl optionally substituted with one or more halogen;R8is absent, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, -alkylene-OH optionally substituted with one or more halogen, -CN, -C(O)-N(R10)2, -C(O)O-alkyl optionally substituted with one or more halogen, or carboxyl;each R10is independently -H, alkyl, or haloalkyl; oralternatively, one of R1, R2a, R3, R4, or R8is substituted with a linker that is optionally linked to a target protein ligand; andwherein at least one of R1, R2a, R3, R4, or R8is not absent or -H.

27. The compound of formula (IA) or pharmaceutically acceptable salt, tautomer, or solvate thereof of claim 26, wherein R1is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, or Ci-Ce alkoxy.

28. The compound of formula (IA) or pharmaceutically acceptable salt, tautomer, or solvate thereof of claim 26 or claim 27, wherein R1is absent, -F, -Cl, -Br, -T, methyl, ethyl, isopropyl, -CF3, methoxy, or ethoxy.

29. The compound of formula (IA) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 26 to 28, wherein R2ais -H, halogen C1-C6 alkyl, or Ci-Ce haloalkyl.

30. The compound of formula (IA) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 26 to 29, wherein R2ais -H, -F, -CF3, methyl, ethyl, or isopropyl.

31. The compound of formula (I A) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 26 to 30, wherein R3is -H, halogen, Ci-Ce alkyl, Ci-Ce or haloalkyl.

32. The compound of formula (IA) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 26 to 31, wherein R3is -H, -F, -CF3, methyl, ethyl, or isopropyl.

33. The compound of formula (IA) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 26 to 32, wherein R4is -H, Ci-Cr, alkyl, or -C(O)-(Ci-Ce alkyl).

34. The compound of formula (IA) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 26 to 33, wherein R4is -H, methyl, ethyl, isopropyl, -C(O)methyl, -C(O)ethyl, or -C(O)isopropyl.

35. The compound of formula (IA) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 26 to 34, wherein R8is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, Ci-C6alkoxy, -(Ci-C6alkylene)-OH, -CN, -C(O)-N(H)(CI-C6alkyl), -C(O)O-(Ci-C6 alkyl), or carboxyl.

36. The compound of formula (TA) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 26 to 35, wherein R8is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, ethoxy, propoxy, -methylene-OH, ethylene-OH, -propylene -OH, -CN, -C(O)-N(H)(methyl), -C(O)-N(H)(ethyl), -C(O)-N(H)(propyl), -C(O)O-(methyl), -C(O)O-(ethyl), -C(O)O-(propyl), or carboxyl.

37. The compound of formula (IA) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 26 to 36, comprising an atropisomer of the compound of formula (IA) or pharmaceutically acceptable salt, tautomer, or solvate thereof, wherein the atropisomer is substantially free of its opposite enantiomer or pharmaceutically acceptable salt, tautomer, or solvate thereof.

38. The compound of formula (IA) or pharmaceutically acceptable salt, tautomer, or solvate thereof of claim 37, wherein the atropisomer is a P isomer.

39. A compound of formula (IB):R4(IB) or a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinR1is absent, halogen, alkyl, haloalkyl, or alkoxy;R2ais alkyl or haloalkyl;R3is -H, halogen, alkyl, or haloalkyl;R4is -H, halogen, alkyl, haloalkyl, or -C(O)-alkyl optionally substituted with one or more halogen;R8is absent, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, -alkylene-OH optionally substituted with one or more halogen, -CN, -C(O)-N(R10)2, -C(O)O-alkyl optionally substituted with one or more halogen, or carboxyl;each R10is independently -H, alkyl, or haloalkyl; oralternatively, one of R1, R2a, R3, R4, or R8is substituted with a linker that is optionally linked to a target protein ligand.

40. The compound of formula (IB) or pharmaceutically acceptable salt, tautomer, or solvate thereof of claim 39, wherein R1is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, or Ci-C6alkoxy.

41. The compound of formula (IB) or pharmaceutically acceptable salt, tautomer, or solvate thereof of claim 39 or claim 40, wherein R1is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, or ethoxy.

42. The compound of formula (IB) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 39 to 41, wherein R2ais Ci-Ce alkyl or Ci-Ce haloalkyl.

43. The compound of formula (IB) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 39 to 42, wherein R2ais -CF3, methyl, ethyl, or isopropyl.

44. The compound of formula (IB) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 39 to 43, wherein R3is -H, halogen, Ci-Ce alkyl, Ci-Ce or haloalkyl.

45. The compound of formula (IB) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 39 to 44, wherein R3is -H, -F, -CF3, methyl, ethyl, or isopropyl.

46. The compound of formula (IB) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 39 to 45, wherein R4is -H, Ci-Ce alkyl, or -C(O)-(Ci-Ce alkyl).

47. The compound of formula (IB) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 39 to 46, wherein R4is -H, methyl, ethyl, isopropyl, -C(O)methyl, -C(O)ethyl, or -C(O)isopropyl.

48. The compound of formula (IB) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 39 to 47, wherein R8is absent, halogen, C1-C6 alkyl, Ci-Cs haloalkyl, Ci-Ce alkoxy, -(Ci-C6alkylene)-OH, -CN, -C(O)-N(H)(Ci-Ce alkyl), -C(O)O-(Ci-Ce alkyl), or carboxyl.

49. The compound of formula (IB) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 39 to 48, wherein R8is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, ethoxy, propoxy, -methylene-OH, ethylene-OH, -propylene -OH, -CN, -C(O)-N(H)(methyl), -C(O)-N(H)(ethyl), -C(O)-N(H)(propyl), -C(O)O-(methyl), -C(O)O-(ethyl), -C(O)O-(propyl), or carboxyl.

50. The compound of formula (IB) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 39 to 49, comprising an atropisomer of the compound of formula (IB) or pharmaceutically acceptable salt, tautomer, or solvate thereof, wherein the atropisomer is substantially free of its opposite enantiomer or pharmaceutically acceptable salt, tautomer, or solvate thereof.

51. The compound of formula (IB) or pharmaceutically acceptable salt, tautomer, or solvate thereof of claim 50, wherein the atropisomer is a P isomer.

52. A compound comprising an atropisomer of formula (IC):HR2a NR (IC) or a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinR1is absent, halogen, alkyl, haloalkyl, or alkoxy;R2ais alkyl or haloalkyl;R8is absent, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, -alkylene-OH optionally substituted with one or more halogen, -CN, -C(O)-N(R10)2, -C(O)O-alkyl optionally substituted with one or more halogen, or carboxyl;each R10is independently -H, alkyl, or haloalkyl; oralternatively, one of R1, R2a, or R8is substituted with a linker that is optionally linked to a target protein ligand; andwherein the compound of formula (IC) or pharmaceutically acceptable salt, tautomer, or solvate thereof is substantially free of its opposite enantiomer or pharmaceutically acceptable salt, tautomer, or solvate thereof.

53. The compound of formula (IC) or pharmaceutically acceptable salt, tautomer, or solvate thereof of claim 52, wherein R1is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, or Ci-Ce alkoxy.

54. The compound of formula (IC) or pharmaceutically acceptable salt, tautomer, or solvate thereof of claim 52 or claim 53, wherein R1is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, or ethoxy.

55. The compound of formula (IC) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 52 to 54, wherein R2ais Ci-Ce alkyl or Ci-Ce haloalkyl.

56. The compound of formula (IC) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 52 to 55, wherein R2ais -CF3, methyl, ethyl, or isopropyl.

57. The compound of formula (IC) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 52 to 56, wherein R8is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, C1-C.6 alkoxy, -(Ci-C.6alkylene)-OH, -CN, -C(0)-N(H)(CI-C6 alkyl), -C(O)O-(Ci-C6 alkyl), or carboxyl.

58. The compound of formula (IA) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 53 to 57, wherein R8is absent, -F, -Cl, -Br, -I, methyl, ethyl, isopropyl, -CF3, methoxy, ethoxy, propoxy, -methylene-OH, ethylene-OH, -propylene -OH, -CN, -C(O)-N(H)(methyl), -C(O)-N(H)(ethyl), -C(O)-N(H)(propyl), -C(O)O-(methyl), -C(O)O-(ethyl), -C(O)O-(propyl), or carboxyl.

59. The compound of formula (IC) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 52 to 58, wherein the atropisomer is a P isomer.

60. The compound of formula (IC) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 52 to 59, having a shorter retention time by chiral separation than its opposite enantiomer or pharmaceutically acceptable salt, tautomer, or solvate thereof.

61. The compound or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 1 to 60, wherein one of R1, R2a, R2b, R3, R4, R5, R6, R8, or R9is substituted with the linker, and the linker is linked to the target protein ligand.H H H H N N N No N o N o N o Ncc& °&a&ca L I I L L0 0 oacceptable salt, tautomer, or solvate thereof.

63. A compound of formula (II):or a pharmaceutically acceptable salt, tautomer, or solvate thereof; whereinR11is H, alkyl, or haloalkyl;R12and R13are each independently absent, halogen, -CN, alkyl, haloalkyl, haloalkenyl, alkynyl, or -alkylene-alkynyl optionally substituted with one or more halogen;R14is cycloalkyl, heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with R17;R15and R16are each independently absent, halogen, alkyl, or haloalkyl;R17is halogen, alkyl, or alkoxy;R18is H, alkyl, or haloalkyl;X1is S(O)n, CH2, or CO;n is 0, 1, or 2; oralternatively, one of R11, R12, R13, R14, R15, R16, R17, or R18is substituted with a linker that is optionally linked to a target protein ligand; andwherein at least one of R12and / or R13is not -CN if X1is S(O)2and R14is a thiophenyl group.

64. The compound of formula (II) or pharmaceutically acceptable salt, tautomer, or solvate thereof of claim 63, wherein R11is H, Ci-Ce alkyl, or Ci-Ce haloalkyl.

65. The compound of formula (II) or pharmaceutically acceptable salt, tautomer, or solvate thereof of claim 63 or claim 64, wherein R11is H, -F, -CF3, methyl, or ethyl.

66. The compound of formula (II) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 63 to 65, wherein R12and R13are each independently absent, halogen, -CN, Ci-Ce alkyl, Ci-Ce haloalkyl, C2-Cehaloalkenyl, C2-Ce alkynyl, -(Ci-C8 alkylene)-(C2-Ce alkynyl).

67. The compound of formula (II) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 63 to 66, wherein R12and R13are each independently absent, -F, -CN, or ethynyl.

68. The compound of formula (II) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 63 to 67, wherein R14is C3-C8 cycloalkyl, heterocyclyl having 5 to 10 ring atoms, (>>-(' 10 aryl, or heteroaryl having 5 to 10 ring atoms, each of which is optionally substituted with R17.

69. The compound of formula (II) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 63 to 68, wherein R14is thiophenyl, phenyl, isoxazolyl, or thiazolyl, each of which is optionally substituted with R17.

70. The compound of formula (II) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 63 to 69, wherein R15and R16are each independently absent, -F, (T-Ce alkyl, or Ci-Ce haloalkyl.

71. The compound of formula (II) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 63 to 70, wherein R15and R16are each independently absent, methyl, ethyl, isopropyl, or -CF3.

72. The compound of formula (II) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 63 to 71, wherein R17is halogen, C1-C6 alkyl, or Ci-G> alkoxy.

73. The compound of formula (II) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 63 to 72, wherein R17is -F, methyl, ethyl, propyl, -CF3, methoxy, ethoxy, or propoxy.

74. The compound of formula (II) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 63 to 73, wherein X1is S(O)2.

75. The compound of formula (II) or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 63 to 74, wherein one of R11, R12, R13, R14, R15, or R16is substituted with a linker that is linked to the target protein ligand.

76. A compound selected from:or a pharmaceutically acceptable salt, tautomer, or solvate.

77. Use of a compound of formula (I), (IA), (IB), (IC), or (II) or a pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 1 to 76 as a molecular glue degrader.

78. A proteolysis targeting chimera (PROTAC) comprising a compound of formula (I), (IA), (IB), (IC), or (II) or a pharmaceutically acceptable salt, tautomer, or solvate thereof, or pharmaceutically acceptable salt, tautomer, or solvate thereof of any of claims 1 to 76.A PROTAC comprising:a compound of formula (I):R4or a pharmaceutically acceptable salt, tautomer, or solvate thereof; whereinR1is absent, halogen, alkyl, haloalkyl, or alkoxy;R2aand R2bare each independently -H, halogen, alkyl, or haloalkyl, or alternatively R2aand R2btogether form a cycloalkyl or heterocyclyl;R3and R5are each independently -H, halogen, alkyl, haloalkyl, or oxo;R4is absent -H, halogen, alkyl, haloalkyl, or -C(O)-alkyl optionally substituted with one or more halogen;R6and R7are each independently -H, halogen, alkyl, or haloalkyl;R8is absent, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, -alkylene-OH optionally substituted with one or more halogen, -CN, -C(O)-N(R10)2, -C(O)O-alkyl optionally substituted with one or more halogen, or carboxyl;R9is -H, halogen, alkyl, haloalkyl, alkoxy optionally substituted with one or more halogen, or -O-alkylene-alkynyl optionally substituted with one or more halogen;each R10is independently -H, alkyl, or haloalkyl;X is N or O; andwherein one of R2a, R2b, R3, R4, R5, R6, R8, R9is substituted with a linker; and a target protein ligand linked to the linker.

80. The PROTAC of claim 79, wherein R1is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, or Ci-Ce alkoxy.

81. The PROTAC of claim 79 or 80, wherein R2aand R2bare each independently -H, halogen Ci-Ce alkyl, or Ci-Ce haloalkyl, or R2aand R2btogether form a C3-C6 cycloalkyl.

82. The PROTAC of any of claims 79 to 81, wherein R3is - is -H, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, or oxo.

83. The PROTAC of any of claims 79 to 82, wherein X is N and R4is -H, Ci-C6alkyl, or -C(O)-(Ci-C6alkyl).

84. The PROTAC of any of claims 79 to 83 wherein R5is -H, halogen, Ci-Ce alkyl, or Ci-Ce haloalkyl.

85. The PROTAC of any of claims 79 to 84, wherein R6is -H, halogen, Ci-Ce alkyl, or Ci-Ce haloalkyl.

86. The PROTAC of any of claims 79 to 85, wherein R7is -H, halogen, Ci-Ce alkyl, or Ci-Ce haloalkyl.

87. The PROTAC of any of claims 79 to 86, wherein R8is absent, halogen, C1-C6 alkyl, Ci-Ce haloalkyl, Ci-Ce alkoxy, -(Ci-Ce alkylene)-OH, -CN, -C(O)-N(H)(Ci-Ce alkyl), -C(O)O-(Ci-Ce alkyl), or carboxyl.

88. The PROTAC of any of claims 79 to 87, wherein R9is -H, Ci-Ce alkyl, Ci-Ce alkoxy, or -O-(Ci-Ce alkylene)- alky nyl.

89. The PROTAC of claim 79, comprising a formula selected from:whereinR1is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, or Ci-Ce alkoxy;R2ais Ci-Ce alkyl, or Ci-Ce haloalkyl;R8is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, Ci-Ce alkoxy, -(Ci-Ce alkylene)-OH, -CN, -C(O)-N(H)(Ci-Ce alkyl), -C(O)O-(Ci-Ce alkyl), or carboxyl;R9is -H, Ci-Ce alkyl, Ci-Ce alkoxy, or -O-(Ci-Ce alkylene)-alkynyl;L is the linker; andTPL is the target protein ligand.

90. The PROTAC of claim 79 comprising a formula selected from:whereinR1is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, or Ci-Ce alkoxy;R2ais Ci-Ce alkyl or Ci-Ce haloalkyl;R8is absent, halogen, Ci-Ce alkyl, Ci-Ce haloalkyl, Ci-Ce alkoxy, -(Ci-Ce alkylene)- OH, -CN, -C(O)-N(H)(CI-C6alkyl), -C(O)O-(Ci-C6alkyl), or carboxyl;L is the linker; andTPL is the target protein ligand.

91. The PROTAC of any of claims 79 to 90, wherein the compound of formula (I) or pharmaceutically acceptable salt, tautomer, or solvate thereof is an atropisomer or pharmaceutically acceptable salt, tautomer, or solvate thereof that is substantially free of its opposite enantiomer or pharmaceutically acceptable salt, tautomer, or solvate thereof.

92. The PROTAC of claim 91, wherein the atropisomer is a P isomer.

93. The PROTAC of any of claims 79 to 92, wherein the linker includes an aliphatic linker, cyclic linker, or polyethylene glycol linker.

94. A PROTAC comprising:a compound of formula (II):or a pharmaceutically acceptable salt, tautomer, or solvate thereof; whereinR11is H, alkyl, or haloalkyl;R12and R13are each independently absent, -CN, alkyl, haloalkyl, haloalkenyl, alkynyl, or -alkylene-alkynyl optionally substituted with one or more halogen;R14is cycloalkyl, heterocyclyl, aryl, or heteroaryl, each of which is optionally substituted with R17;R15and R16are each independently absent, halogen, alkyl, or haloalkyl;R17is halogen, alkyl, or alkoxy;R18is -H, alkyl, or haloalkyl;X1is S(O)„, CH2, or CO;n is 0, 1, or 2;wherein one R11, R12, R13, R14, R15, R16, R17, or R18is substituted with a linker and a target protein ligand is linked to the linker.

95. The PROTAC of claim 94, wherein R11is H, Ci-Ce alkyl, Ci-Ce haloalky 1.

96. The PROTAC of claim 94 or 95, wherein R11is H, methyl, or ethyl.

97. The PROTAC of any of claims 94 to 96, wherein R12and R13are each independently absent, -CN, Ci-Ce alkyl, Ci-Cehaloalkyl, C2-Cehaloalkenyl, C2-C6 alkynyl, -(C1-C6 alkylene)-(C2-C6 alkynyl).

98. The PROTAC of any of claims 94 to 97, wherein R14is Cj-Cs cycloalkyl, heterocyclyl having 5 to 10 ring atoms, Ce-Cio aryl, or heteroaryl having 5 to 10 ring atoms, each of which is optionally substituted with R17.

99. The PROTAC of any of claims 94 to 98, wherein R14is thiophenyl, phenyl, or thiazolyl.

100. The PROTAC of any of claims 94 to 99, wherein X1is S(O)2.

101. The PROTAC of any of claims 94 to 100, wherein R15and R16are each independently absent, halogen, Ci-Ce alkyl, or Ci-Ce haloalkyl.

102. The PROTAC of claim 94, comprising the formula:pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinR14is C.3-Cs cycloalkyl, heterocyclyl having 5 to 10 ring atoms, C6-C10 aryl, or heteroaryl having 5 to 10 ring atoms, each of which is optionally substituted with R17;R15and R16arc each independently absent, halogen, Ci-Ce alkyl, or C i-Ce haloalkyl;R17is halogen, alkyl, or alkoxy;L1is the linker; andTPL is the target protein ligand.

103. The PROTAC of claim 94, comprising the formula:CNCNor a pharmaceutically acceptable salt, tautomer, or solvate thereof;whereinL1is the linker; andTPL is the target protein ligand.

104. The PROTAC of any of claims 94 to 103, wherein the linker includes an aliphatic linker, a cyclic linker, or a polyethylene glycol linker.

105. The PROTAC of any of claims 79 to 104, wherein the target protein ligand binds to a target protein selected from B7.1 and B7, TINFRlm, TNFR2, NADPH oxidase, BclIBax and other partners in the apoptosis pathway, C5a receptor, HMG-CoA reductase, PDE V phosphodiesterase type, PDE IV phosphodiesterase type 4, PDE I, PDEII, PDEIII, squalene cyclase inhibitor, CXCR1, CXCR2, nitric oxide (NO) synthase, cyclo-oxygenase 1, cyclo-oxygenase 2, alpha-synuclein, HIV capsid protein, VP40, BFT-bromodomain protein, condensate-forming oncogenic fusion proteins including nucleoporin-containing fusion proteins (e.g., NUP98-NSD1, NUP98-KDM5A, NUP98-PHF23; DEK-NUP214, NUP214-ABL1, SET-NUP214, etc.), EML4-ALK, and BRD4-NUT, or oligomeric proteins involved in inflammatory signaling including NLRP3, RIPK3, RIPK1, MLKL, and GSDM3, 5HT receptors, dopamine receptors, G Proteins, Gq, histamine receptors, 5-lipoxygenase, tryptase serine protease, thymidylate synthase, purine nucleoside phosphorylase, GAPDH trypanosomal, glycogen phosphorylase, Carbonic anhydrase, chemokine receptors, JAW STAT, RXR and similar, HIV 1 protease, HIV 1 integrase, influenza, neuramimidase, hepatitis B reverse transcriptase, sodium channel, multi drug resistance (MDR), protein P-glycoprotein (and MRP), tyrosine kinases, CD23, CD124, tyrosine kinase p56 lek, CD4, CD5, IL-2 receptor, IL-1 receptor, TNF-aR, ICAM1, Cat-i- channels, VCAM, VLA-4 integrin, selectins, CD40 / CD40L, newokinins and receptors, inosine monophosphate dehydrogenase, p38 MAP Kinase, Ras / Raf / ME / ERK pathway, interleukin- 1 converting enzyme, caspase, HCV, NS3 protease, HCV NS3 RNA helicase, glycinamide ribonucleotide formyl transferase, rhinovirus 3C protease, herpes simplex virus- 1 (HSV-I), protease, cytomegalovirus (CMV) protease, poly (ADP-ribose) polymerase, cyclin dependent kinases, vascular endothelial growth factor, c-Kit, TGFP activated kinase 1, mammalian target of rapamycin, SHP2, androgen receptor, oxytocin receptor, microsomal transfer protein inhibitor, bile acid transport inhibitor, 5 alpha reductase inhibitors, angiotensin 11, glycine receptor, noradrenaline reuptake receptor, estrogen receptor, estrogen related receptors, focal adhesion kinase, Src, endothelin receptors, neuropeptide Y and receptor, adenosine receptors, adenosine kinase and AMP deaminase, purinergic receptors (P2Y1, P2Y2, P2Y4, P2Y6, P2X1-7), famesyltransferases, geranylgeranyl transferase, TrkA a receptor for NGF, amyloid, tau or pathological tau aggregates, tyrosine kinase Flk-IIKDR, vitronectin receptor, integrin receptor, Her-21 neu, telomerase inhibition, cytosolic phospholipaseA2, or EGF receptor tyrosine kinase.

106. The PROTAC of any of claims 79 to 104, wherein the target protein ligand binds to a target protein selected from tau, amyloid, alpha-synuclein, HIV capsid protein, VP40, condensate-forming oncogenic fusion proteins including nucleoporin-containing fusion proteins (e.g., NUP98-NSD1, NUP98-KDM5A, NUP98-PHF23; DEK-NUP214, NUP214-ABL1, SET-NUP214, etc.) EML4-ALK, and BRD4-NUT, or oligomeric proteins involved in inflammatory signaling including NLRP3, RIPK3, RIPK1, MLKL, and GSDM3.

107. A PROTAC having a formula selected from:; or a pharmaceutically acceptable salt, tautomer, or solvate thereof.

108. A PROTAC having a formula selected from:acceptable salt, tautomer, or solvate thereof.

109. The PROTAC of any of claims 79 to 108, promoting targeted protein degradation, such as pathological protein degradation or pathological aggregated protein degradation.

110. The PROTAC of any of claims 79 to 109 for use in targeted protein degradation, such as pathological protein degradation or pathological aggregated protein degradation.

111. The PROTAC of any of claims 79 to 107 for targeted degradation of pathogenic tau.

112. The PROTAC of any of claims 79 to 106 or 108 for use in treating cancer.

113. The PROTAC of any of claims 79 to 111 for use in aggregated protein degradation for central nervous system diseases, neurodegenerative diseases, and other diseases, such as Alzheimer’s disease and frontotemporal dementia.

114. The PROTAC of any of claims 80 to 106, for use in treating asthma, multiple sclerosis, cancer, ciliopathics, cleft palate, diabetes, heart disease, hypertension, inflammatory bowel disease, mental retardation, mood disorder, obesity, refractive error, infertility, Angelman syndrome, Canavan disease, Coeliac disease, Charcot-Marie-Toothdisease, Cystic fibrosis, Duchenne muscular dystrophy, Haemochromatosis, Haemophilia, Klinefelter's syndrome, Neurofibromatosis, Phenylketonuria, Polycystic kidney disease, (PKD1) or 4 (PKD2) Prader-Willi syndrome, Sickle-cell disease, Tay-Sachs disease, Turner syndrome.

115. The PROTAC of any of claims 79 to 106, for use in treating Alzheimer's disease, Amyotrophic lateral sclerosis (Lou Gehrig's disease), Anorexia nervosa, Anxiety disorder, Atherosclerosis, Attention deficit hyperactivity disorder, Autism, Bipolar disorder, Chronic fatigue syndrome, Chronic obstructive pulmonary disease, Crohn's disease, Coronary heart disease, Dementia, Depression, Diabetes mellitus type 1, Diabetes mellitus type 2, Epilepsy, Guillain-Barre syndrome, Irritable bowel syndrome, Lupus, Metabolic syndrome, Multiple sclerosis, Myocardial infarction, Obesity, Obscssivc-compulsivc disorder, Panic disorder, Parkinson's disease, Psoriasis, Rheumatoid arthritis, Sarcoidosis, Schizophrenia, Stroke, Thromboangiitis obliterans, Tourette syndrome, Vasculitis.

116. The PROTAC of any of claims 79 to 106 for use in treating aceruloplasminemia, Achondrogenesis type II, achondroplasia, Acrocephaly, Gaucher disease type 2, acute intermittent porphyria, Canavan disease, Adenomatous Polyposis Coli, ALA dehydratase deficiency, adenylosuccinate lyase deficiency, Adrenogenital syndrome, Adrenoleukodystrophy, ALA-D porphyria, ALA dehydratase deficiency, Alkaptonuria, Alexander disease, Alkaptonuric ochronosis, alpha 1 -antitrypsin deficiency, alpha- 1 proteinase inhibitor, emphysema, amyotrophic lateral sclerosis, Alstrom syndrome, Alexander disease, Amelogenesis imperfecta, ALA dehydratase deficiency, Anderson-Fabry disease, androgen insensitivity syndrome, Anemia, Angiokeratoma Corporis Diffusum, Angiomatosis retinae (von HippeLLindau disease), Apert syndrome, Arachnodactyly (Marfan syndrome), Stickler syndrome, Arthrochalasis multiplex congenital (Ehlers-Danlos syndrome arthrochalasia type), ataxia telangiectasia, Rett syndrome, primary pulmonary hypertension, Sandhoff disease, neurofibromatosis type II, Beare-Stevenson cutis gyrata syndrome, Mediterranean fever, familial, Benjamin syndrome, beta-thalassemia, Bilateral Acoustic Neurofibromatosis (neurofibromatosis type II), factor V Leiden thrombophilia, Bloch-Sulzberger syndrome (incontinentia pigmenti), Bloom syndrome, X-linked sideroblastic anemia, Bonnevie-Ullrich syndrome (Turner syndrome), Bourneville disease(tuberous sclerosis), prion disease, Birt-Hogg-Dube syndrome, Brittle bone disease (osteogenesis imperfecta), Broad Thumb-Hallux syndrome (Rubinstein-Taybi syndrome), Bronze Diabetes / Bronzed Cirrhosis (hemochromatosis), Bulbospinal muscular atrophy (Kennedy's disease), Burger-Grutz syndrome (lipoprotein lipase deficiency), CGD Chronic granulomatous disorder, Campomelic dysplasia, biotinidase deficiency, Cardiomyopathy (Noonan syndrome), Cri du chat, CAVD (congenital absence of the vas deferens), Caylor cardiofacial syndrome (CBAVD), CEP (congenital erythropoietic porphyria), cystic fibrosis, congenital hypothyroidism, Chondrodystrophy syndrome (achondroplasia), otospondylomegaepiphyseal dysplasia, Lesch-Nyhan syndrome, galactosemia, Ehlers-Danlos syndrome, Thanatophoric dysplasia, Coffin-Lowry syndrome, Cockayne syndrome, (familial adenomatous polyposis), Congenital erythropoietic porphyria. Congenital heart disease, Mcthcmoglobincmia / Congcnital mcthacmoglobinacmia, achondroplasia, X-linkcd sideroblastic anemia, Connective tissue disease, Conotruncal anomaly face syndrome, Cooley's Anemia (beta-thalassemia). Copper storage disease (Wilson's disease), Copper transport disease (Menkes disease), hereditary coproporphyria, Cowden syndrome, Craniofacial dysarthrosis (Crouzon syndrome), Creutzfeldt- Jakob disease (prion disease), Cockayne syndrome, Cowden syndrome, Curschmann-Batten-Steinert syndrome (myotonic dystrophy), Beare-Stevenson cutis gyrata syndrome, primary hyperoxaluria, spondyloepimetaphyseal dysplasia (Strudwick type), muscular dystrophy, Duchenne and Becker types (DBMD), Usher syndrome, Degenerative nerve diseases including de Grouchy syndrome and Dejerine-Sottas syndrome, developmental disabilities, distal spinal muscular atrophy, type V, androgen insensitivity syndrome, Diffuse Globoid Body Sclerosis (Krabbe disease), Di George's syndrome, Dihydrotestosterone receptor deficiency, androgen insensitivity syndrome, Down syndrome, Dwarfism, erythropoietic protoporphyria, Erythroid 5 -aminolevulinate synthetase deficiency, Erythropoietic porphyria, erythropoietic protoporphyria, erythropoietic uroporphyria, Friedreich's ataxia, familial paroxysmal polyserositis, porphyria cutanea tarda, familial pressure sensitive neuropathy, primary pulmonary hypertension (PPH), Fibrocystic disease of the pancreas, fragile X syndrome, galactosemia, genetic brain disorders, Giant cell hepatitis (Neonatal hemochromatosis), Gronblad-Strandberg syndrome (pseudoxanthoma elasticum), Gunther disease (congenital erythropoietic porphyria), haemochromatosis, Hallgren syndrome, sickle cell anemia, hemophilia, hepatoerythropoietic porphyria (HEP), Hippel-Lindau disease (von Hippel-Lindau disease). Huntington's disease, Hutchinson-Gilford progeria syndrome (progeria), Hyperandrogenism, Hypochondroplasia, Hypochromic anemia, Immune system disorders, including X-linked severe combined immunodeficiency, Insley-Astley syndrome, Jackson-Weiss syndrome, Joubert syndrome, Lesch-Nyhan syndrome, Jackson-Weiss syndrome, Kidney diseases, including hyperoxaluria, Klinefelter's syndrome, Kniest dysplasia, Lacunar dementia, Langer-Saldino achondrogenesis, ataxia telangiectasia, Lynch syndrome, Lysylhydroxylase deficiency, Machado-Joseph disease, Metabolic disorders, including Kniest dysplasia, Marfan syndrome, Movement disorders, Mowat- Wilson syndrome, cystic fibrosis, Muenke syndrome, Multiple neurofibromatosis, Nance-Insley syndrome, Nance-Sweeney chondrodysplasia, Niemann-Pick disease, Noack syndrome (Pfeiffer syndrome), Osler-Weber-Rendu disease, Peutz-Jeghers syndrome, Polycystic kidney disease, polyostotic fibrous dysplasia (McCunc-Albright syndrome), Pcutz-Jcghcrs syndrome, Pradcr-Labhart-Willi syndrome, hemochromatosis, primary hyperuricemia syndrome (Lesch-Nyhan syndrome), primary pulmonary hypertension, primary senile degenerative dementia, prion disease, progeria (Hutchinson Gilford Progeria Syndrome), progressive chorea, chronic hereditary (Huntington) (Huntington's disease), progressive muscular atrophy, spinal muscular atrophy, propionic acidemia, protoporphyria, proximal myotonic dystrophy, pulmonary arterial hypertension, PXE (pseudoxanthoma elasticum), Rb (retinoblastoma), Recklinghausen disease (neurofibromatosis type I), Recurrent polyserositis, Retinal disorders, Retinoblastoma, Rett syndrome, RFALS type 3, Ricker syndrome, Riley-Day syndrome, Roussy-Levy syndrome, severe achondroplasia with developmental delay and acanthosis nigricans (SADDAN), Li-Fraumeni syndrome, sarcoma, breast, leukemia, and adrenal gland (SBLA) syndrome, sclerosis tuberose (tuberous sclerosis), SDAT, SED congenital (spondyloepiphyseal dysplasia congenita), SED Strudwick (spondyloepimetaphyseal dysplasia, Strudwick type), SEDc (spondyloepiphyseal dysplasia congenita), SEMD, Strudwick type (spondyloepimetaphyseal dysplasia, Strudwick type), Shprintzen syndrome, Skin pigmentation disorders, Smiih-Lcmli-Opiiz syndrome, South-African genetic porphyria (variegate porphyria), infantile-onset ascending hereditary spastic paralysis, Speech and communication disorders, sphingolipidosis, Tay-Sachs disease, spinocerebellar ataxia, Stickler syndrome, stroke, androgen insensitivity syndrome, tetrahydrobiopterin deficiency, beta-thalassemia, Thyroid disease Tomaculous neuropathy (hereditary neuropathy with liability to pressure palsies) Treacher Collins syndrome, TripleX syndrome (triple X syndrome), Trisomy 21 (Down syndrome), Trisomy X, VHL syndrome (von Hippel-Lindau disease), Vision impairment and blindness (Alstrom syndrome), Vrolik disease, Waardenburg syndrome, Warburg Sjo Hedelius Syndrome, Weissenbacher-Zweymuller syndrome, Wolf-Hirschhorn syndrome, Wolff Periodic disease, Weissenbacher-Zweymiiller syndrome and Xcruderma pigmentosum.

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