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US 10,662,248 B2Granted
De novo binding domain containing polypeptides and uses thereof
AssigneesSubdomain, LLC, Arcellx, Inc.
InventorsDavid William Lafleur, David M. Hilbert
Life of this patent
Estimating the term…
Abstract
Provided herein are de novo binding domain containing polypeptides (DBDpp) that specifically bind a target of interest. Nucleic acids encoding the DBDpp, and vectors and host cells containing the nucleic acids are also provided. Libraries of DBDpp, methods of producing and screening such libraries and the DBDpp identified from such libraries and screens are also encompassed. Methods of making and using the DBDpp are additionally provided. Such uses include, without limitation, affinity purification, and diagnostic and therapeutic applications.
Claims28
A polypeptide for binding a target of interest, the polypeptide comprising an amino acid sequence selected from the group consisting of:
- MGSWX5X6FKX9X10LAX13IKX16X17LEALGGSEAELAX30FEX33X34IAX37FEX40X41LQX44YKGKGNPEVEALRKEAAAIRDELQAYRHN (SEQ ID NO:2),
- MGSWAEFKQRLAAIKTRLEALGGSEAELAAFX32X33EIX36AFX39X40ELX43AYKGKGNPEVEALX57X58EAX61AIX64X65ELX68AYRHN (SEQ ID NO:3),
- MGSWX5EFX8X9RLX12AIX15X16RLX19ALGGSEAELAAFEKEIAAFESELQAYKGKGNPEVEX55LRX58X59AAX62IRX65X66LQAYRHN (SEQ ID NO:4),
- MGSWX5X6FKX9X10LAX13IKX16X17LEALGGSEAELAAFX32X33EIX36AFX39X40ELX43AYKGKGNPEVEX55LRX58X59AAX62IRX65X66LQAYRHN (SEQ ID NO:5),
- MGSWX5EFX8X9RLX12AIX15X16RLX19ALGGSEAELAX30FEX33X34IAX37FEX40X41LQX44YKGKGNPEVEALX57X58EAX61AIX64X65ELX68AYRHN (SEQ ID NO:6),
- MGSWX5X6FKX9X10LAX13IKX16X17LEALZ1EAELAX28FEX31X32IAX35FEX38X39LQX42YZ2NPEVEALRKEAAAIRDELQAYRHN (SEQ ID NO:7),
- MGSWAEFKQRLAAIKTRLEALZ1EAELAAFX30X31EIX34AFX37X38ELX41AYZ2NPEVEALX52X53EAX56AIX59X60ELX63AYRHN (SEQ ID NO:8),
- MGSWX5EFX8X9RLX12AIX15X16RLX19ALZ1EAELAAFEKEIAAFESELQAYZ2NPEVEX50LRX53X54AAX57IRX60X61LQAYRHN (SEQ ID NO:9),
- MGSWX5X6FKX9X10LAX13IKX16X17LEALZ1EAELAAFX30X31EIX34AFX37X38ELX41AYZ2NPEVEX50LRX53X54AAX57IRX60X61LQAYRHN (SEQ ID NO:10),
- MGSWX5EFX8X9RLX12AIX15X16RLX19ALZ1EAELAX28FEX31X32IAX35FEX38X39LQX42YZ2NPEVEALX52X53EAX56AIX59X60ELX63AYRHN (SEQ ID NO:
- 11), and combinations thereof;
- wherein Xn is a natural or non-natural amino acid, wherein each Xn is the same or different natural or non-natural amino acid,
- wherein Z1 and Z2 comprise between about 2 to about 30 natural or non-natural amino acids,
- wherein the amino acid sequence differs from SEQ ID NO:
- 1 and does not comprise SEQ ID NO:
- 50, and
- wherein the polypeptide specifically binds a target of interest.
14 more elements
A de novo binding domain polypeptide (DBDpp), wherein the DBDpp immunospecifically binds to a protein comprising amino acids 19-305 of CD123 (SEQ ID NO:
- 187) or a protein that is at least 95% identical to CD123,
- wherein the DBDpp comprises an amino acid sequence selected from the group consisting of:
- MGSWX5X6FKX9X10LAX13IKX16X17LEALGGSEAELAX30FEX33X34IAX37FEX40X41LQX44YKGKGNPEVEALRKEAAAIRDELQAYRHN (SEQ ID NO:2),
- MGSWAEFKQRLAAIKTRLEALGGSEAELAAFX32X33EIX36AFX39X40ELX43AYKGKGNPEVEALX57X58EAX61AIX64X65ELX68AYRHN (SEQ ID NO:3),
- MGSWX5EFX8X9RLX12AIX15X16RLX19ALGGSEAELAAFEKEIAAFESELQAYKGKGNPEVEX55LRX58X59AAX62IRX65X66LQAYRHN (SEQ ID NO:4),
- MGSWX5X6FKX9X10LAX13IKX16X17LEALGGSEAELAAFX32X33EIX36AFX39X40ELX43AYKGKGNPEVEX55LRX58X59AAX62IRX65X66LQAYRHN (SEQ ID NO:5),
- MGSWX5EFX8X9RLX12AIX15X16RLX19ALGGSEAELAX30FEX33X34IAX37FEX40X41LQX44YKGKGNPEVEALX57X58EAX61AIX64X65ELX68AYRHN (SEQ ID NO:6),
- MGSWX5X6FKX9X10LAX13IKX16X17LEALZ1EAELAX28FEX31X32IAX35FEX38X39LQX42YZ2NPEVEALRKEAAAIRDELQAYRHN (SEQ ID NO:7),
- MGSWAEFKQRLAAIKTRLEALZ1EAELAAFX30X31EIX34AFX37X38ELX41AYZ2NPEVEALX52X53EAX56AIX59X60ELX63AYRHN (SEQ ID NO:8),
- MGSWX5EFX8X9RLX12AIX15X16RLX19ALZ1EAELAAFEKEIAAFESELQAYZ2NPEVEX50LRX53X54AAX57IRX60X61LQAYRHN (SEQ ID NO:9),
- MGSWX5X6FKX9X10LAX13IKX16X17LEALZ1EAELAAFX3X31EIX34AFX37X38ELX41AYZ2NPEVEX50LRX53X54AAX57IRX60X61LQAYRHN (SEQ ID NO:10),
- MGSWX5EFX8X9RLX12AIX15X16RLX19ALZ1EAELAX28FEX31X32IAX35FEX38X39LQX42YZ2NPEVEALX52X53EAX56AIX59X60ELX63AYRHN (SEQ ID NO:11), and combinations thereof;
- wherein Xn is a natural or non-natural amino acid, wherein each Xn is the same or different natural or non-natural amino acid, and wherein Xn is not cysteine or proline,
- wherein Z1 and Z2 comprise between about 2 to about 30 natural or non-natural amino acids, and wherein the amino acid sequence differs from SEQ ID NO:
- 1 and does not comprise SEQ ID NO:
- 50.
13 more elements
A cell comprising a nucleic acid sequence comprising a sequence encoding a chimeric antigen receptor (CAR), wherein the CAR comprises (i) an antigen binding domain comprising a de novo binding domain polypeptide (DBDpp), (ii) a transmembrane domain, and (iii) a signaling domain,
- wherein the DBDpp comprises an amino acid sequence selected from the group consisting of:
- MGSWX5X6FKX9X10LAX13IKX16X17LEALGGSEAELAX30FEX33X34IAX37FEX40X41LQX44YKGKGNPEVEALRKEAAAIRDELQAYRHN (SEQ ID NO:2),
- MGSWAEFKQRLAAIKTRLEALGGSEAELAAFX32X33EIX36AFX39X40ELX43AYKGKGNPEVEALX57X58EAX61AIX64X65ELX68AYRHN (SEQ ID NO:3),
- MGSWX5EFX8X9RLX12AIX15X16RLX19ALGGSEAELAAFEKEIAAFESELQAYKGKGNPEVEX55LRX58X59AAX62IRX65X66LQAYRHN (SEQ ID NO:4),
- MGSWX5X6FKX9X10LAX13IKX16X17LEALGGSEAELAAFX32X33EIX36AFX39X40ELX43AYKGKGNPEVEX55LRX58X59AAX62IRX65X66LQAYRHN (SEQ ID NO:5),
- MGSWX5EFX8X9RLX12AIX15X16RLX19ALGGSEAELAX30FEX33X34IAX37FEX40X41LQX44YKGKGNPEVEALX57X58EAX61AIX64X65ELX68AYRHN (SEQ ID NO:6),
- MGSWX5X6FKX9X10LAX13IKX16X17LEALZ1EAELAX28FEX31X32IAX35FEX38X39LQX42YZ2NPEVEALRKEAAAIRDELQAYRHN (SEQ ID NO:7),
- MGSWAEFKQRLAAIKTRLEALZ1EAELAAFX30X31EIX34AFX37X38ELX41AYZ2NPEVEALX52X53EAX56AIX59X60ELX63AYRHN (SEQ ID NO:8),
- MGSWX5EFX8X9RLX12AIX15X16RLX19ALZ1EAELAAFEKEIAAFESELQAYZ2NPEVEX50LRX53X54AAX57IRX60X61LQAYRHN (SEQ ID NO:9),
- MGSWX5X6FKX9X10LAX13IKX16X17LEALZ1EAELAAFX30X31EIX34AFX37X38ELX41AYZ2NPEVEX50LRX53X54AAX57IRX60X61LQAYRHN (SEQ ID NO:10),
- MGSWX5EFX8X9RLX12AIX15X16RLX19ALZ1EAELAX28FEX31X32IAX35FEX38X39LQX42YZ2NPEVEALX52X53EAX56AIX59X60ELX63AYRHN (SEQ ID NO:11), and combinations thereof;
- wherein Xn is a natural or non-natural amino acid, wherein each Xn is the same or different natural or non-natural amino acid, and wherein Xn is not cysteine or proline,
- wherein Z1 and Z2 comprise between about 2 to about 30 natural or non-natural amino acids, and
- wherein the amino acid sequence differs from SEQ ID NO:
- 1 and does not comprise SEQ ID NO:
- 50.
13 more elements
A method of treating a subject having cancer, the method comprising:
- administering to the subject an immune cell comprising a chimeric antigen receptor (CAR), wherein the CAR comprises:
- a) a target binding domain comprising an amino acid sequence selected from the group consisting of:
- MGSWX5X6FKX9X10LAX13IKX16X17LEALGGSEAELAX30FEX33X34IAX37FEX4X41LQX44YKGKGNPEVEALRKEAAAIRDELQAYRHN (SEQ ID NO:2),
- MGSWAEFKQRLAAIKTRLEALGGSEAELAAFX32X33EIX36AFX39X40ELX43AYKGKGNPEVEALX57X58EAX61AIX64X65ELX68AYRHN (SEQ ID NO:3),
- MGSWX5EFX8X9RLX12AIX15X16RLX19ALGGSEAELAAFEKEIAAFESELQAYKGKGNPEVEX55LRX58X59AAX62IRX65X66LQAYRHN (SEQ ID NO:4),
- MGSWX5X6FKX9X10LAX13IKX16X17LEALGGSEAELAAFX32X33EIX36AFX39X40ELX43AYKGKGNPEVEX55LRX58X59AAX62IRX65X66LQAYRHN (SEQ ID NO:5),
- MGSWX5EFX8X9RLX12AIX15X16RLX19ALGGSEAELAX30FEX33X34IAX37FEX40X41LQX44YKGKGNPEVEALX57X58EAX61AIX64X65ELX68AYRHN (SEQ ID NO:6),
- MGSWX5X6FKX9X10LAX13IKX16X17LEALZ1EAELAX28FEX31X32IAX35FEX38X39LQX42YZ2NPEVEALRKEAAAIRDELQAYRHN (SEQ ID NO:7),
- MGSWAEFKQRLAAIKTRLEALZ1EAELAAFX30X31EIX34AFX37X38ELX41AYZ2NPEVEALX52X53EAX56AIX59X60ELX63AYRHN (SEQ ID NO:8),
- MGSWX5EFX8X9RLX12AIX15X16RLX19ALZ1EAELAAFEKEIAAFESELQAYZ2NPEVEX50LRX53X54AAX57IRX60X61LQAYRHN (SEQ ID NO:9),
- MGSWX5X6FKX9X10LAX13IKX16X17LEALZ1EAELAAFX3X31EIX34AFX37X38ELX41AYZ2NPEVEX50LRX53X54AAX57IRX60X61LQAYRHN (SEQ ID NO:10),
- MGSWX5EFX8X9RLX12AIX15X16RLX19ALZ1EAELAX28FEX31X32IAX35FEX38X39LQX42YZ2NPEVEALX52X53EAX56AIX59X60ELX63AYRHN (SEQ ID NO:
- 11), and combinations thereof;
- wherein Xn is a natural or non-natural amino acid, wherein each Xn is the same or different natural or non-natural amino acid, and wherein Xn is not cysteine or proline,
- wherein Z1 and Z2 comprise between about 2 to about 30 natural or non-natural amino acids,
- wherein the amino acid sequence differs from SEQ ID NO:
- 1 and does not comprise SEQ ID NO:
- 50, and
- wherein the target binding domain specifically binds a target of interest expressed by a cancer cell,
- b) a transmembrane domain, and
- c) an intracellular domain, wherein the intracellular domain comprises a signaling domain,
- wherein, upon administration to a subject having cancer, the target binding domain specifically binds to the target of interest expressed by a cancer cell, and
- wherein the binding of the target of interest induces the immune cell to generate cytotoxic signals that result in cytotoxic effects on the cancer cell, thereby treating the cancer.
20 more elements
Dependencies are inferred from the claim text.