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US 2021/0171998 A1Published application
Lysine conjugated immunoglobulins
AssigneeEisai R&D Management Co., Ltd.
InventorsJared Spidel, Earl Albone
Life of this patent
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Abstract
Provided herein are conjugated immunoglobulins and methods for generating conjugated immunoglobulins using a microbial transglutaminase.
Claims27
A conjugated immunoglobulin comprising an immunoglobulin or an antigen-binding portion thereof, and an acyl donor substrate, wherein
- a) the immunoglobulin, or antigen-binding portion thereof, comprises an engineered lysine residue, wherein the engineered lysine residue is a natural amino acid residue which has been mutated to a lysine residue,
- wherein the natural amino acid residue which has been mutated to a lysine residue is selected from the group consisting of:
- Threonine 135 (T135K), Serine 136 (S136K), Leucine 193 (L193K), Aspartic acid 221 (D221K), Threonine 223 (T223K), Histidine 224 (H224K), Threonine 225 (T225K), Methionine 252 (M252K), Asparagine 297 (N297K), and Proline 445 (P445K) on a heavy chain of the immunoglobulin, or antigen-binding portion thereof,
- Leucine 201 (L201K) and Serine 202 (S202K) on a kappa light chain of the immunoglobulin, or antigen-binding portion thereof, and
- Glutamic acid 213 (E213K) on a lambda light chain of the immunoglobulin, or antigen-binding portion thereof;
- wherein the amino acid residues are numbered according to the EU numbering system;
- b) the acyl donor substrate comprises a glutamine residue,
- wherein the acyl donor substrate is according to one of Formulae (I) or (II):
- (Z)m-Gln-(L)n-(Y) (I)
- (Y)-(L)n-Gln-(Z)m (II)
- wherein
- Z is a carboxylbenzyloxy (CBZ) group or an amino acid residue;
- Gln is a glutamine amino acid residue;
- each L is independently a straight or branched linker from 1 to 20 carbon atoms, wherein one or more of the carbon atoms may be optionally and independently replaced with a nitrogen, oxygen or sulfur atom, and wherein each carbon and nitrogen atom may be optionally substituted;
- or each L is optionally and independently an amino acid residue;
- m is an integer from 0 to 5;
- n is an integer from 0 to 5;
- and
- Y is a therapeutic agent or a diagnostic agent
- wherein the engineered lysine residue of the immunoglobulin, or antigen-binding portion thereof, is conjugated to the glutamine residue of the acyl donor substrate.
17 more elements
A method for generating a conjugated immunoglobulin, the method comprising:
- contacting an immunoglobulin, or antigen-binding portion thereof, with a microbial transglutaminase and an acyl donor substrate,
- a) wherein the immunoglobulin, or antigen-binding portion thereof, comprises an engineered lysine residue, wherein the engineered lysine residue is a natural amino acid residue which has been mutated to a lysine residue,
- wherein the natural amino acid residue which has been mutated to a lysine residue is selected from the group consisting of:
- Threonine 135 (T135K), Serine 136 (S136K), Leucine 193 (L193K), Aspartic acid 221 (D221K), Threonine 223 (T223K), Histidine 224 (H224K), Threonine 225 (T225K), Methionine 252 (M252K), Asparagine 297 (N297K), and Proline 445 (P445K) on a heavy chain of the immunoglobulin, or antigen-binding portion thereof,
- Leucine 201 (L201K) and Serine 202 (S202K) on a kappa light chain of the immunoglobulin, or antigen-binding portion thereof, and
- Glutamic acid 213 (E213K) on a lambda light chain of the immunoglobulin, or antigen-binding portion thereof;
- wherein the amino acid residues are numbered according to the EU numbering system;
- b) wherein the acyl donor substrate comprises a glutamine residue, and wherein the acyl donor substrate is according to one of Formulae (I) or (II):
- (Z)m-Gln-(L)n-(Y) (I)
- (Y)-(L)n-Gln-(Z)m (II)
- wherein
- Z is a carboxylbenzyloxy (CBZ) group or an amino acid residue;
- Gln is a glutamine amino acid residue;
- each L is independently a straight or branched linker from 1 to 20 carbon atoms, wherein one or more of the carbon atoms may be optionally and independently replaced with a nitrogen, oxygen or sulfur atom, and wherein each carbon and nitrogen atom may be optionally substituted;
- or each L is optionally and independently an amino acid residue;
- m is an integer from 0 to 5;
- n is an integer from 0 to 5;
- and
- Y is a therapeutic agent or a diagnostic agent
- wherein the microbial transglutaminase conjugates the engineered lysine residue of the immunoglobulin, or antigen-binding portion thereof, to the glutamine residue of the acyl donor substrate, thereby generating the conjugated immunoglobulin.
17 more elements
Dependencies are inferred from the claim text.