US 11,753,669 B2
Lysine conjugated immunoglobulins
Jared Spidel, Downingtown, PA (US); and Earl Albone, Blue Bell, PA (US)
Assigned to Eisai R&D Management Co., Ltd., Tokyo (JP)
Filed by Eisai R&D Management Co., Ltd., Tokyo (JP)
Filed on Jan. 27, 2021, as Appl. No. 17/159,655.
Application 17/159,655 is a division of application No. 15/662,981, filed on Jul. 28, 2017, granted, now 10,941,431.
Application 15/662,981 is a continuation of application No. PCT/JP2017/021672, filed on Jun. 12, 2017.
Claims priority of provisional application 62/348,410, filed on Jun. 10, 2016.
Prior Publication US 2021/0171998 A1, Jun. 10, 2021
Int. Cl. C07K 16/00 (2006.01); C12P 21/02 (2006.01); A61K 47/68 (2017.01)
CPC C12P 21/02 (2013.01) [A61K 47/68 (2017.08); A61K 47/6803 (2017.08); A61K 47/6811 (2017.08); C07K 16/00 (2013.01); C12Y 203/02013 (2013.01); C07K 2317/40 (2013.01); C07K 2317/41 (2013.01); C07K 2317/51 (2013.01); C07K 2317/515 (2013.01); C07K 2317/52 (2013.01); C07K 2317/522 (2013.01); C07K 2317/524 (2013.01); C07K 2317/53 (2013.01); C07K 2317/55 (2013.01)] 27 Claims
OG exemplary drawing
 
1. 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.