Covalent attachment of ligands to nucleophilic proteins guided by non-covalent binding
AssigneeSudhir Paul
InventorsSudhir Paul, Yasuhiro Nishiyama
Life of this patent
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Abstract
A covalently reactive ligand analogue (CAL) of formula (1): wherein, L1 . . . Lx . . . Lm are components defining a ligand determinant, Lx is a component unit of the ligand determinant selected from the group consisting of an amino acid residue, sugar residue, a fatty acid residue and a nucleotide, L′ is a functional group of Lx, Y″ is atom, covalent bond or linker, Y′ is an optional charged or neutral group Y is a covalently reactive electrophilic group that reacts specifically with a receptor that binds to said ligand determinant, and n is an integer from 1 to 1000 m is an integer from 1 to 30.
Claims61
A covalently reactive ligand analogue (CAL) of formula (1):
- wherein,
- L1 . . . Lx . . . Lm are components defining a ligand determinant,
- Lx is a component unit of the ligand determinant selected from the group consisting of an amino acid residue, sugar residue, a fatty acid residue and a nucleotide,
- L′ is a functional group of Lx,
- Y″ is atom, covalent bond or linker,
- Y′ is an optional charged or neutral group
- Y is a covalently reactive electrophilic group that reacts specifically with a receptor that binds to said ligand determinant, and
- n is an integer from 1 to 1000
- m is an integer from 1 to 30.
A method for activating or inactivating a nucleophilic receptor (NuR), comprising:
- contacting a covalently reactive ligand analogue (CAL) of formula (1):
- wherein,
- L1 . . . Lx . . . Lm are components defining a ligand determinant,
- Lx is a component unit of the ligand determinant selected from the group consisting of an amino acid residue, sugar residue, a fatty acid residue and a nucleotide,
- L′ is a functional group of Lx,
- Y″ is atom, covalent bond or linker,
- Y′ is an optional charged or neutral group
- Y is a covalently reactive electrophilic group that reacts specifically with a receptor that binds to said ligand determinant, and
- n is an integer from 1 to 1000
- m is an integer from 1 to 30;
- with a nucleophilic receptor that reacts specifically with the ligand determinant of said CAL.
A method for activating or inactivating a nucleophilic receptor (NuR) produced by a microorganism, comprising:
- contacting a covalently reactive ligand analogue (CAL) of formula (1):
- wherein,
- L1 . . . Lx . . . Lm are components defining a ligand determinant,
- Lx is a component unit of the ligand determinant selected from the group consisting of an amino acid residue, sugar residue, a fatty acid residue and a nucleotide,
- L′ is a functional group of Lx,
- Y″ is atom, covalent bond or linker,
- Y′ is an optional charged or neutral group
- Y is a covalently reactive electrophilic group that reacts specifically with a receptor that binds to said ligand determinant, and
- n is an integer from 1 to 1000
- m is an integer from 1 to 30;
- with a nucleophilic receptor produced by a microorganism, that reacts specifically with the ligand determinant of said CAL.
A method for agonism or antagonism of nucleophilic receptors (NuRs) expressed on cellular surfaces, comprising:
- contacting a covalently reactive ligand analogue (CAL) of formula (1):
- wherein,
- L1 . . . Lx . . . Lm are components defining a ligand determinant,
- Lx is a component unit of the ligand determinant selected from the group consisting of an amino acid residue, sugar residue, a fatty acid residue and a nucleotide,
- L′ is a functional group of Lx,
- Y″ is atom, covalent bond or linker,
- Y′ is an optional charged or neutral group
- Y is a covalently reactive electrophilic group that reacts specifically with a receptor that binds to said ligand determinant, and
- n is an integer from 1 to 1000
- m is an integer from 1 to 30;
- with a cell containing a NuR on its surface that reacts specifically with the ligand determinant of said CAL, resulting in activation or inactivation of cellular signal transducing system.
A method for inducing growth arrest or death of cells, comprising:
- contacting a covalently reactive ligand analogue (CAL) of formula (1) to which a cytostatic or cytotoxic agent is optionally attached by a covalent bond:
- wherein,
- L1 . . . Lx . . . Lm are components defining a ligand determinant,
- Lx is a component unit of the ligand determinant selected from the group consisting of an amino acid residue, sugar residue, a fatty acid residue and a nucleotide,
- L′ is a functional group of Lx,
- Y″ is atom, covalent bond or linker,
- Y′ is an optional charged or neutral group
- Y is a covalently reactive electrophilic group that reacts specifically with a receptor that binds to said ligand determinant, and
- n is an integer from 1 to 1000
- m is an integer from 1 to 30;
- with lymphocytes that express NuRs which react specifically with the ligand determinant of said CAL, resulting in cellular growth arrest or death.
A method for preparing self-assembled biomolecules, comprising:
- subjecting a CAL of formula (1) to conditions which promote the formation of self-assembled multiple units, optionally incorporating one or more cofactor in the assembly.
A method of vaccination, comprising:
- administering to subject in need thereof an effective amount of a pharmaceutical composition containing a covalently reactive ligand analogue (CAL) of formula (1):
- wherein,
- L1 . . . Lx . . . Lm are components defining a ligand determinant,
- Lx is a component unit of the ligand determinant selected from the group consisting of an amino acid residue, sugar residue, a fatty acid residue and a nucleotide,
- L′ is a functional group of Lx,
- Y″ is atom, covalent bond or linker,
- Y′ is an optional charged or neutral group
- Y is a covalently reactive electrophilic group that reacts specifically with a receptor that binds to said ligand determinant, and
- n is an integer from 1 to 1000
- m is an integer from 1 to 30.
A method for passive immunotherapy, comprising:
- administering to a subject in need thereof an effective amount of a pharmaceutical composition containing antibodies that bind to a covalently reactive ligand analogue (CAL) of formula (1):
- wherein,
- L1 . . . Lx . . . Lm are components defining a ligand determinant,
- Lx is a component unit of the ligand determinant selected from the group consisting of an amino acid residue, sugar residue, a fatty acid residue and a nucleotide,
- L′ is a functional group of Lx,
- Y″ is atom, covalent bond or linker,
- Y′ is an optional charged or neutral group
- Y is a covalently reactive electrophilic group that reacts specifically with a receptor that binds to said ligand determinant, and
- n is an integer from 1 to 1000
- m is an integer from 1 to 30.
A method for imaging, comprising:
- exposing cells or tissues which express a NuR on the cell surfaces to a detectably labeled derivative of a CAL of formula (1), and detecting the label.
A method for producing receptors or enzymes having improved ligand binding or enzymatic activity, comprising:
- subjecting a receptor or an enzyme to directed evolution or a mutation process, and isolating a receptor or enzyme with improved ligand binding or enzymatic activity, by contacting it with a CAL of formula (1).
An immunoassay for detection of antibodies or antigens, comprising:
- contacting an immobilized CAL of formula (1) or a detectably labeled CAL of formula (1).
Dependencies are inferred from the claim text.