US 11,723,983 B2
Conjugates of a glycoprotein or a glycan with a toxic payload
Tero Satomaa, Helsinki (FI); Jari Helin, Rajamäki (FI); and Filip S. Ekholm, Porvoo (FI)
Assigned to GLYKOS FINLAND OY, Helsinki (FI)
Filed by GLYKOS FINLAND OY, Helsinki (FI)
Filed on Mar. 23, 2021, as Appl. No. 17/209,356.
Application 17/209,356 is a continuation of application No. 15/704,026, filed on Sep. 14, 2017, granted, now 10,973,922.
Application 15/704,026 is a continuation of application No. 14/888,545, abandoned, previously published as PCT/FI2014/050322, filed on May 2, 2014.
Claims priority of application No. 20135451 (FI), filed on May 2, 2013; and application No. 20136020 (FI), filed on Oct. 14, 2013.
Prior Publication US 2021/0252158 A1, Aug. 19, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. A61K 47/61 (2017.01); C07K 9/00 (2006.01); A61K 47/68 (2017.01); A61K 47/54 (2017.01); A61K 47/64 (2017.01); A61K 31/404 (2006.01); A61K 31/704 (2006.01); A61K 38/08 (2019.01); C07K 16/28 (2006.01); C07K 16/30 (2006.01); C07K 16/32 (2006.01)
CPC A61K 47/61 (2017.08) [A61K 31/404 (2013.01); A61K 31/704 (2013.01); A61K 38/08 (2013.01); A61K 47/549 (2017.08); A61K 47/64 (2017.08); A61K 47/6803 (2017.08); A61K 47/6807 (2017.08); A61K 47/6817 (2017.08); A61K 47/6849 (2017.08); A61K 47/6851 (2017.08); A61K 47/6869 (2017.08); A61K 47/6889 (2017.08); C07K 9/001 (2013.01); C07K 16/2863 (2013.01); C07K 16/3069 (2013.01); C07K 16/32 (2013.01); C07K 2317/24 (2013.01); C07K 2317/90 (2013.01); Y02A 50/30 (2018.01)] 4 Claims
 
1. A method for preparing a glycoprotein (Gp)-toxic payload molecule conjugate of Formula I:
[D-L-G]n-Gp   Formula (I)
wherein:
Gp is an antibody comprising a hybrid-type N-glycan having an acceptor site and a GlcNAc residue bound by a β-N linkage to an asparagine;
n is an integer from 1 to about 20;
D is a toxic payload molecule;
L is a linker group covalently joining G to D; and
G is a saccharide structure of Formula II

OG Complex Work Unit Chemistry
and wherein:
R is a glycosidic bond to the 4-position of the GlcNAc residue bound by a β-N linkage to an asparagine;
X1 is H;
X2 is NHCOCH2-triazole, wherein the triazole comprises a bond to L;
X3 and X4 are each OH;
X5 is CH2OH; and,
the anomeric structure of G is in β-D-galacto configuration;
and wherein the method consists of the steps of:
contacting Gp with an endoglycosidase selected from EndoS49 to produce a EndoS49-deglycosylated Gp;
reacting a donor molecule with the acceptor site of the EndoS49-deglycosylated GP in the presence of a glycosyltransferase to form a G-Gp conjugate; and
reacting the G component of the G-Gp conjugate with a compound of Formula XIV
D-L-L″   Formula (XIV)
wherein:
D is the toxic payload molecule;
L is the linker group covalently joining L″ to D; and
L″ is an alkyne for forming the triazole moiety in X2;
wherein the N-glycan of Gp, after the contacting with the endoglycosidase, is selected from the structure of Formula IV:

OG Complex Work Unit Chemistry
wherein (β-N-Asn) is a β-N linkage to an asparagine and y is 0 or 1.