# US 2020/0399356 A1 · Synthetic antibodies to bax and uses thereof > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2020/0399356 A1 - Document ID: US20200399356A1 - Status: Published application - Kind: A1 - Application number: 16/988,947 - Filed: 2020-08-10 - Published: 2020-12-24 - Earliest claimed: 2015-09-10 - Assignees: Albert Einstein College of Medicine; The Governing Council of the University of Toronto - Applicants: Albert Einstein College of Medicine; The Governing Council of the University of Toronto - Inventors: Evripidis Gavathiotis; Jonathan R. Lai; Sachdev Sidhu - Web page: https://patentgrep.com/patent/US20200399356A1 ## CPC - C07K 16/18: against material from animals or humans - A61K 9/127: Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant - A61K 47/6843: {the antibody targeting a material from animals or humans} - G01N 33/575: for cancer - C07K 16/005: {constructed by phage libraries} - C07K 2317/10: characterized by their source of isolation or production - C07K 2317/24: containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered - C07K 2317/55: Fab or Fab' - C07K 2317/565: Complementarity determining region [CDR] - C07K 2317/622: Single chain antibody (scFv) - C07K 2317/76: Antagonist effect on antigen, e.g. neutralization or inhibition of binding - C07K 2317/92: Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value - G01N 2500/04: Screening involving studying the effect of compounds C directly on molecule A (e.g. C are potential ligands for a receptor A, or potential substrates for an enzyme A) - G01N 2510/00: Detection of programmed cell death, i.e. apoptosis - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) ## Abstract Synthetic fragment antigen-binding (Fab) antibodies are disclosed that bind to an N-terminal activation site of BCL-2-associated X-protein (BAX) and inhibit BAX activation. Also disclosed are methods of using the Fabs for measuring inactive monomeric BAX levels, screening for small molecules that bind to an N-terminal activation site of BAX, inhibiting apoptotic cell death, and predicting the ability of a cancer therapy to promote apoptotic cell death. ## Claims ### Claim 1 A synthetic fragment antigen-binding (Fab) antibody that binds to an N-terminal activation site of BCL-2-associated X-protein (BAX) and inhibits BAX activation, wherein the synthetic fragment antigen-binding (Fab) antibody is selected from the group consisting of: 3H4 Fab, having a light chain comprising a CDR L1, a CDRL2, and a CDRL3 region comprising the amino acid sequence QSWYYSYPI (SEQ ID NO:25), a CDRH1 region comprising the amino sequence LSYSSM (SEQ ID NO 26), a CDRH2 region comprising the amino acid sequence SISSYYSYTS (SEQ ID NO:27), and a CDRH3 region comprising the amino acid sequence RYYGYGGGID (SEQ ID NO-28); 2A6 Fab, having a light chain comprising a CDR L1, a CDRL2, and a CDRL3 region comprising the amino acid sequence QSAGGYPLI (SEQ ID NO:29), a CDRH1 region comprising the amino sequence IYYSSM (SEQ ID NO-30), a CDRH2 region comprising the amino acid sequence SISPYSSYTS (SEQ ID NO:31), and a CDRH3 region comprising the amino acid sequence RSFGYCWAFD (SEQ ID NO:32); 3H1 Fab, having a light chain comprising a CDR L1, a CDRL2, and a CDRL3 region comprising the amino acid sequence QHSYPI (SEQ ID NO:33), a CDRH1 region comprising the amino sequence ISYSSI (SEQ ID NO:34), a CDRH2 region comprising the amino acid sequence SIYSYSGSTY (SEQ ID NO 35), and a CDRH3 region comprising the amino acid sequence RYGAMD (SEQ ID NO:36); 2A2 Fab, having a light chain comprising a CDR L1, a CDRL2, and a CDRL3 region comprising the amino acid sequence QYYYPI (SEQ ID NO:37), a CDRH1 region comprising the amino sequence ISSSSI (SEQ ID NO:38), a CDRH2 region comprising the amino acid sequence SIYSYYGYTY (SEQ ID NO:39), and a CDRH3 region comprising the amino acid sequence RYSAMD (SEQ ID NO:40); 3G3 Fab, having a light chain comprising a CDR L1, a CDRL2, and a CDRL3 region comprising the amino acid sequence QGAWSGGHLI (SEQ ID NO 41), a CDRH1 region comprising the amino sequence LSYSSM (SEQ ID NO:42), a CDRH2 region comprising the amino acid sequence YISPYYGYTY (SEQ ID NO:43), and a CDRH3 region comprising the amino acid sequence RGWAYYYGYWGPSGLD (SEQ ID NO:44); 2D5 Fab, having a light chain comprising a CDR L1, a CDRL2, and a CDRL3 region comprising the amino acid sequence QWGYSHSHLI (SEQ ID NO:45), a CDRH1 region comprising the amino sequence ISYSS (SEQ ID NO:46), a CDRH2 region comprising the amino acid sequence SISPYYGSTY (SEQ ID NO:47), and a CDR-13 region comprising the amino acid sequence RSHFGALD (SEQ ID NO:48), 2D2 Fab, having a light chain comprising a CDR L1, a CDRL2, and a CDRL3 region comprising the amino acid sequence QSYYWVSPF (SEQ ID NO:49), a CDRH1 region comprising the amino sequence LYYSSI (SEQ ID NO:50), a CDRH2 region comprising the amino acid sequence SIYPYSGSTY (SEQ ID NO:51), and a CDRH3 region comprising the amino acid sequence RSYGYAMD (SEQ ID NO:52); and 2A5 Fab, having a light chain comprising a CDR L1, a CDRL2, and a CDRL3 region comprising the amino acid sequence QYHYWYYPI (SEQ ID NO 53), a CDRH1region comprising the amino sequence ISYYSM (SEQ ID NO:54), a CDRH2 region comprising the amino acid sequence SISPYYGSTY (SEQ ID NO:55), and a CDRH3 region comprising the amino acid sequence RAGAMD (SEQ ID NO:56). ### Claim 2 (depends on 1) The synthetic Fab antibody of claim 1, wherein the antibody maintains BAX in its inactive, monomeric form. ### Claim 3 (depends on 1) The synthetic Fab antibody of claim 1, wherein the antibody binds to residues of helices α1 and α6, and to residues of α1-α2 loop of BAX. ### Claim 4 (depends on 1) The synthetic Fab antibody of claim 1, wherein the antibody binds to BAX with a half-maximal binding (EC50) affinity of 2 nM-70 nM. ### Claim 5 (depends on 1) The synthetic Fab antibody of claim 1, wherein the antibody blocks the interaction of BAX with a BAX activating partner. ### Claim 6 (depends on 5) The synthetic Fab antibody of claim 5, wherein the BAX activating partner is selected from the group consisting of tBID, PUMA, BIM and NOXA. ### Claim 7 (depends on 1) The synthetic Fab antibody of claim 1, wherein the synthetic Fab antibody is conjugated to a therapeutic agent. ### Claim 8 (depends on 1) The synthetic Fab antibody of claim 1, wherein the synthetic Fab antibody is conjugated to an agent that facilitates transport across a cell membrane. ### Claim 9 (depends on 1) The synthetic Fab antibody of claim 1, wherein the synthetic Fab antibody is labeled with a fluorescent label or a radioactive label.