US 12,240,790 B2
Radiolabeling of polypeptides
Vadim Dudkin, Chalfont, PA (US); Shalom Goldberg, Merion Station, PA (US); Joseph Erhardt, Sellersville, PA (US); Rhys Salter, Doylestown, PA (US); and Theresa McDevitt, Warminster, PA (US)
Assigned to Janssen Biotech, Inc., Horsham, PA (US)
Appl. No. 16/955,094
Filed by Janssen Biotech, Inc., Horsham, PA (US)
PCT Filed Dec. 17, 2018, PCT No. PCT/US2018/065913
§ 371(c)(1), (2) Date Jun. 18, 2020,
PCT Pub. No. WO2019/125982, PCT Pub. Date Jun. 27, 2019.
Claims priority of provisional application 62/599,830, filed on Dec. 18, 2017.
Prior Publication US 2021/0017099 A1, Jan. 21, 2021
Int. Cl. A61K 39/00 (2006.01); A61K 51/10 (2006.01); A61P 35/00 (2006.01); C07B 59/00 (2006.01); C07K 16/30 (2006.01)
CPC C07B 59/008 (2013.01) [A61K 51/1072 (2013.01); A61K 51/1096 (2013.01); A61P 35/00 (2018.01); C07K 16/3069 (2013.01); A61K 2039/505 (2013.01); C07B 2200/05 (2013.01); C07K 2317/41 (2013.01)] 23 Claims
 
1. A method of labeling a polypeptide with a radiometal ion, the method comprising:
a. providing an antibody or an antigen binding fragment thereof that is covalently linked to a first click reaction partner, wherein the first click reaction partner comprises an azide, and wherein the antibody or antigen binding fragment thereof that is covalently linked to a first click reaction partner is obtained by a site-specific incorporation of the first click reaction partner through a reaction with an azide-labeled sugar or an azido amine;
b. providing a radiocomplex comprising the radiometal ion and a chelating moiety coordinated with the radiometal ion, wherein the chelating moiety comprises a chelant covalently linked to a second click reaction partner, wherein the second click reaction partner comprises a strained alkyne group; and
c. contacting the antibody or antigen binding fragment thereof that is covalently linked to a first click reaction partner with the radiocomplex, wherein the contacting is performed without a copper catalyst, to allow the first click reaction partner to react with the second click reaction partner to thereby label the antibody or an antigen binding fragment thereof with the radiometal ion;
wherein the chelant comprises a macrocycle having the structure of formula (I):

OG Complex Work Unit Chemistry
wherein each of R1, R2, R3 and R4 is independently CHQCO2X, wherein
Q is independently hydrogen, C1-C4 alkyl or (C1-C2 alkyl) phenyl, and
X is independently hydrogen, benzyl, C1-C4 alkyl; and
Z is (CH2)n Y, wherein
n is 1-10, and
Y is a linker that covalently links the second click reaction partner and the chelant, and wherein Y is a bond, C(O)—(CH2)m—C(O), (CH2)p—C(O)—NH—(CH2)q—C(O), (O—CH2—CH2)r—O, (CH2)p[C(O)—NH—(CH2)q]t, (CH2)p—S—S—(CH2)q, or valine-citrulline-PAB, wherein p=0-4, q=1-4, r=1-4, t=1-3, m=1-4;
alternatively, Z is hydrogen; and, when Z is hydrogen then
each of R1, R2, R3 and R4 is independently CHQCO2X, wherein
Q is a linker that covalently links the second click reaction partner and the chelant, and
wherein Q is a bond, C(O)—(CH2)m—C(O), (CH2)p—C(O)—NH—(CH2)q—C(O), (O—CH2—CH2)r—O, (CH2)p[C(O)—NH—(CH2)q]t, (CH2)p—S—S—(CH2)q, or valine-citrulline-PAB,
wherein p=0-4, q=1-4, r=1-4, t=1-3, and
X is independently hydrogen, benzyl, or C1-C4 alkyl;
or alternatively, the chelant comprises deferoxamine-YY, wherein YY is a linker that covalently links the second click reaction partner and the chelant, and wherein YY is a bond, C(O)—(CH2)m—C(O), (CH2)p—C(O)—NH—(CH2)q—C(O), (O—CH2—CH2)r—O, (CH2)p[C(O)—NH—(CH2)q]t, (CH2)p—S—S—(CH2)q, or valine-citrulline-PAB, wherein p=0-4, q=1-4, r=1-4, t=1-3.