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CN-121985961-A - TRIM21 binding molecules

CN121985961ACN 121985961 ACN121985961 ACN 121985961ACN-121985961-A

Abstract

The present invention relates to binding molecules comprising a TRIM21 binding moiety and a protein binding moiety for selectively degrading a target protein. The invention also relates to compositions comprising these molecules and their use in therapy. The invention also relates to TRIM21 binding molecules and uses thereof.

Inventors

  • LEO JAMES
  • DAVID FRANCE
  • DEAN CLIFT
  • Jacob Luptak

Assignees

  • 英国研究与创新署

Dates

Publication Date
20260505
Application Date
20240719
Priority Date
20230721

Claims (20)

  1. 1. A compound of formula (I) A-L-B (I) Or a pharmaceutically acceptable salt thereof, wherein: A is a moiety that binds to TRIM21, wherein a has a molecular weight of 1000 Da or less; L is a linker, and B is a moiety that binds to a target protein, wherein the target protein is capable of comprising a first form that provides a plurality of binding sites for B.
  2. 2. The compound of claim 1, wherein the target protein is capable of forming an oligomeric species.
  3. 3. The compound of claim 1 or claim 2, wherein the compound is configured to recruit TRIM21 such that one or more oligomeric forms of the target protein are selectively degradable by TRIM 21.
  4. 4. A compound according to any one of claims 1-3, wherein a binds to: (a) PRYSPRY domain of TRIM21 (SEQ ID NO: 1), and/or (B) One or more of the residues selected from W381, W383, D355 and F369, L370, L371, F450, Y328, H368, S447, M330, D452, R364, Q395, Y393, E389 and K387 of TRIM21, preferably one or more residues selected from W381, W383 and D355.
  5. 5. The compound of any one of the preceding claims, wherein a competes with IgG for binding to TRIM21.
  6. 6. The compound of any one of the preceding claims, wherein a is not an antibody.
  7. 7. A compound according to any one of the preceding claims, wherein the compound has a K d value for TRIM21 in the range 0.1nM to 1000 μΜ, preferably 10nM to 100 μΜ.
  8. 8. The compound of any one of the preceding claims, wherein a has the structure of formula (II) (II), Wherein: R 1 and R 2 are each independently selected from hydrogen and C 1-2 alkyl, or one of R 1 and R 2 is hydrogen and the other is selected from an amino acid residue and is a site of covalent attachment to L; r 3 is selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl, wherein R 3 optionally comprises a site of covalent attachment to L; r 4 is hydrogen, halogen or C 1-2 alkyl; R 5 is hydrogen or C 1-2 alkyl and R 6 is optionally substituted alkyl, or R 5 and R 6 combine to form an optionally substituted heterocycle; r 7 is Sites or covalent attachments to L ; R 8 is hydrogen or C 1-2 alkyl; r 9 is optionally substituted alkyl; R 14 and R 15 are each independently selected from hydrogen and C 1-2 alkyl, and Of the structure of R 7 Indicating the site of covalent attachment to L; provided that a comprises a site of covalent attachment to L.
  9. 9. The compound of any one of the preceding claims, wherein a has the structure of formula (IIa), (IIb) or (IIc): (IIa)、 (IIb) or (IIc) Wherein: Each of which is Independently selected from: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 And ; R 1 and R 2 are each independently selected from hydrogen and C 1-2 alkyl, or one of R 1 and R 2 is hydrogen and the other is an amino acid residue; R 3 is selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl; r 4 is hydrogen, halogen or C 1-2 alkyl; each R 5 is independently hydrogen or C 1-2 alkyl; R 6 is optionally substituted alkyl; Or when R 5 and R 6 are attached to adjacent atoms, R 5 and R 6 combine to form an optionally substituted heterocycle; R 8 is hydrogen or C 1-2 alkyl; r 9 is optionally substituted alkyl; r 18 and R 19 are each independently selected from hydrogen and C 1-2 alkyl, and R 20 is optionally substituted alkyl.
  10. 10. The compound according to claim 8 or 9, wherein R 1 、R 2 and R 4 are hydrogen.
  11. 11. The compound of any one of claims 8-10, wherein R 3 is selected from optionally substituted aryl and optionally substituted heteroaryl, optionally wherein R 3 is selected from furan and optionally substituted phenyl.
  12. 12. The compound of any one of claims 8, 10 and 11, wherein R 7 is Wherein: (a) R 9 is selected from the group consisting of-alkylene-optionally substituted aryl and-alkylene-optionally substituted heteroaryl, optionally wherein R 9 is selected from the group consisting of-CH 2 -optionally substituted aryl and-CH 2 -optionally substituted heteroaryl, or (B) R 9 is selected from 、 、 、 And Wherein R 9 is in the structure of Indicating the site of covalent attachment to the remainder of R 7 .
  13. 13. A compound according to any one of claims 9 to 11, wherein: (a) R 9 is selected from the group consisting of-alkylene-optionally substituted aryl and-alkylene-optionally substituted heteroaryl, optionally wherein R 9 is selected from the group consisting of-CH 2 -optionally substituted aryl and-CH 2 -optionally substituted heteroaryl, or (B) R 9 is selected from 、 、 、 And 。
  14. 14. The compound of any one of claims 8-13, wherein R 5 and R 6 combine to form a 5-membered heterocycle optionally substituted with hydroxy, nitro, cyano, halogen, -NR 10 R 11 , optionally substituted alkyl, optionally substituted alkoxy, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted-O-cycloalkyl, optionally substituted-O-heterocyclyl, optionally substituted-O-aryl, or optionally substituted-O-heteroaryl, Wherein R 10 and R 11 are each independently H, optionally substituted alkyl or optionally substituted cycloalkyl.
  15. 15. The compound of any one of the preceding claims, wherein a has the structure of formula (III) (III), Wherein: Each of rings E, F, C and D is independently selected from optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl, and In the structure of formula (III) Indicating the site of covalent attachment to L.
  16. 16. The compound of claim 15, wherein ring C is selected from aryl and heteroaryl, wherein the aryl or heteroaryl is substituted with aryl or heteroaryl.
  17. 17. The compound of any one of the preceding claims, wherein a is selected from: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 And , Wherein A is of the structure Indicating the site of covalent attachment to L.
  18. 18. The compound of claim 17, wherein a is selected from: 、 And , Wherein A is of the structure Indicating the site of covalent attachment to L.
  19. 19. The compound of any one of the preceding claims, wherein the target protein is selected from the group consisting of: (a) Proteins forming aggregates involved in neurodegeneration, preferably wherein the protein is selected from tau, synuclein, amyloid β, islet amyloid polypeptide (IAPP), serum Amyloid A (SAA), prion protein, misfolded transthyretin (TTR), TAR DNA binding protein 43 (TDP-43), NOTCH3 receptor, mutated cystatin C, medin, dipeptide repeat protein and FUS protein, e.g. a protein selected from tau, synuclein, amyloid β, prion protein, TAR DNA binding protein 43 (TDP-43), dipeptide repeat protein and FUS protein, preferably wherein the target protein is tau; (b) A protein having amplified repeat elements, preferably wherein the protein is a mutated huntingtin; (c) A protein involved in a signal transduction pathway in inflammation and/or cancer, preferably wherein the protein is selected from Myd88, IRAK4, IRAK1, IRAK2, TRAF6, NLRP3, RIPK3 and ASC, preferably wherein the protein is selected from Myd88, IRAK4, IRAK1, IRAK2, TRAF6, NLRP3 and ASC; (d) An oligomer-forming viral protein, preferably wherein the viral protein is selected from the group consisting of CoV2 and influenza virus nucleoprotein, or (E) Proteins selected from bromodomain-containing protein 4 (BRD 4), mutated gelsolin, misfolded rhodopsin and atrial natriuretic peptide.
  20. 20. The compound of any one of the preceding claims, wherein B is selected from PI-2014, FDDNP, AV680 (T808), GTP-1, THK523, THK5105, PBB3, AV1451 (T807), THK5117, THK5351, N-methyllansoprazole, (E) -6-iodo-2-styryl-1H-benzo [ d ] imidazole, RO6958948, PI-2620, JNJ-067, APN-1607 (PM-PBB 3), MK-6240, and methylene blue.

Description

TRIM21 binding molecules Technical Field The present invention relates to binding molecules comprising a TRIM21 binding moiety and a protein binding moiety for selectively degrading a target protein. The invention also relates to compositions comprising these molecules and their use in therapy. The invention also relates to TRIM21 binding molecules and uses thereof. Background Small molecules called "protein degradation targeting chimeras" (PROTAC) can be used to deplete target proteins. PROTAC molecules typically comprise a ligand, such as a small molecule inhibitor of the protein of interest, covalently linked to a ligand of E3 ubiquitin ligase. Upon binding of the small molecule inhibitor to the target protein, PROTAC molecules recruit the E3 ubiquitin ligase, which ubiquitinates the target protein, targeting it for subsequent degradation. PROTAC molecules are considered potential therapies for the treatment of a variety of diseases, allowing clinicians to specifically target and degrade specific proteins. However, although PROTAC molecules are selective for the target protein, PROTAC molecules may not distinguish between different forms of the same protein, and thus may result in degradation of the non-pathogenic form of the protein. Thus, there is a need for molecules that are capable of targeting pathogenic forms of proteins. Such molecules will enable more targeted therapies. The use of more targeted protein degradation as a therapeutic strategy can minimize off-target effects of drugs and avoid or reduce systemic drug exposure. Summary of The Invention The present invention relates to bifunctional molecules comprising a TRIM21 binding moiety and a protein targeting moiety. Since the compounds of the present invention are capable of aggregating TRIM21 in the vicinity of the target protein, selective degradation of a specific form of the target protein can be achieved. Accordingly, in a first aspect the present invention provides a compound of formula (I) A-L-B (I) Or a pharmaceutically acceptable salt thereof, wherein: A is a moiety that binds to TRIM21, wherein a has a molecular weight of 1000 Da or less; L is a linker, and B is the portion that binds to the target protein. The target protein is a protein that may comprise a first form (e.g., a pathogenic form) that provides multiple binding sites for B. The target protein may be a protein capable of forming an oligomeric species (oligomeric species). When in oligomeric form, multiple binding sites are provided for B. In a second aspect the invention provides a pharmaceutical composition comprising a compound of the first aspect of the invention, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. In a third aspect the invention provides an in vitro method of selectively degrading an oligomeric or pathogenic form of a target protein, the method comprising contacting the target protein with TRIM21 and a compound according to the first aspect of the invention or a pharmaceutically acceptable salt thereof. In a fourth aspect the present invention provides a compound having the structure of formula (IV). A fifth aspect of the invention relates to compounds having the structure of formula (V). Other aspects and embodiments of the invention are described below and in the drawings. All preferred features of the second and subsequent aspects of the invention are as in the first aspect mutatis mutandis. Drawings FIG. 1 shows the thermal stabilization of Trim21 PRYSPRY (10. Mu.M) in the presence of compounds 36, 37 and 38 (100. Mu.M) compared to DMSO only control. FIG. 2 shows the quenching of the natural fluorescence of Trim21 PRYSPRY after ligand binding-Compound 36 (black circles), compound 37 (dark grey squares) and Compound 38 (grey triangles). The data were fitted using a four parameter combined model. Fig. 3 shows fluorescence polarization shift measurements. White circles show titration of human IgG-Fc against Alexa-488 labeled Trim21 PRYSPRY. For displacement experiments, 0.5 μm IgG-Fc was used with titration of competitor compounds 36 (black circles), 37 (dark grey squares) and 38 (grey triangles). FIG. 4 shows a lane view (a) and quantification (b) of TRIM21 protein levels normalized to Hsp60 protein levels and DMSO conditions after treatment of RPE-1 cells with DMSO or 2.5 μM compound 37 for 18h and harvesting whole cell extracts for capillary-based immunoblotting. Significance was based on student t-test (two-tailed). FIG. 5 shows a schematic representation of an assay for TRIM 21-mediated degradation. Expression of NbGFP-Fc antibody constructs resulted in recruitment of TRIM21 to H2B-mEGFP, resulting in degradation of the H2B-mEGFP proteasome. Compound 37 competes with NbGFP-Fc for binding to TRIM21 PRYSPRY, thereby inhibiting TRIM21 recruitment and H2B-mEGFP degradation. FIG. 6 shows the H2B-mEGFP fluorescence values normalized to NbGFP-Fc H433A (Fc mutant incapable of binding TRIM21 PRYSPRY; no degradation