US 12,467,928 B2
N-terminal modifier agents and binders for treating and analyzing peptides
Eric Okerberg, San Diego, CA (US); Stephen Verespy, III, San Diego, CA (US); Jason C. Klima, San Diego, CA (US); Soumya Ganguly, San Diego, CA (US); Zachary Miles, San Diego, CA (US); Jason Duarte Jacintho, San Diego, CA (US); Aaron Wise, San Diego, CA (US); and Wadud Bhuiya, San Diego, CA (US)
Filed by Encodia, Inc., San Diego, CA (US)
Filed on Jul. 5, 2024, as Appl. No. 18/764,943.
Claims priority of provisional application 63/525,347, filed on Jul. 6, 2023.
Prior Publication US 2024/0377408 A1, Nov. 14, 2024
Int. Cl. C40B 70/00 (2006.01); C40B 40/10 (2006.01); G01N 33/68 (2006.01)
CPC G01N 33/6842 (2013.01) [C40B 40/10 (2013.01); C40B 70/00 (2013.01); G01N 33/6845 (2013.01)] 28 Claims
 
22. A composition comprising:
(a) a solid support, wherein the solid support comprises a plurality of peptide molecules attached to the solid support, wherein each of the plurality of peptide molecules comprise a modified N-terminal amino acid (NTAA) residue generated by modifying a peptide molecule with an N-terminal modifier agent, wherein the modified NTAA residue is capable of coordinating or chelating a metal cation, and
the N-terminal modifier agent is selected from the group consisting of:

OG Complex Work Unit Chemistry
wherein M is a metal cation binding group that comprises sulfonamide, hydroxamic acid, sulfamate, or sulfamide; the group

OG Complex Work Unit Chemistry
is a 5 or 6 membered aromatic ring containing up to three heteroatoms selected from N, O, and S as ring members, and is optionally substituted by R; R represents one or two optional substituents selected from the group consisting of F, Cl, CH3, CF2H, CF3, OH, OCH3, OCF3, NH2, N(CH3)2, NO2, SCH3, SO2CH3, CH2OH, B(OH)2, CN, CONH2, CO2H, and CONHCH3; LG is OH, ORQ, or OCC, each RQ is independently aryl or heteroaryl, each of which is optionally substituted with one or more groups selected from halo, nitro, cyano, sulfonate, carboxylate, alkylsulfonyl, and N of heteroaryl is optionally oxidized; or RQ can be —C(═O)R or —C(═O)—OR; CC is a cationic counterion; X is one of the following: O, S, Se, or NH,
wherein the modified NTAA residue of each of the plurality of peptide molecules comprises one of the structures selected from the group consisting of:

OG Complex Work Unit Chemistry
wherein AA1 is a side chain of the modified NTAA residue and PP is a peptide structure except for the modified NTAA residue; and
(b) an engineered metalloprotein binder that specifically binds to the modified NTAA residue of a peptide molecule of the plurality of peptide molecules, wherein the engineered metalloprotein binder comprises an amino acid sequence X1-C/H/D/E-X2-C/H/D/E-X3-C/H/D/E-X4, wherein C/H/D/E is any single amino acid residue independently selected from the group consisting of amino acid residues C (Cys), H (His), D (Asp) and E (Glu); X1, X2, X3 and X4 are each any amino acid sequence comprising between 0 and 500 amino acid residues in length, and wherein the amino acid sequence X1-C/H/D/E-X2-C/H/D/E-X3-C/H/D/E-X4 chelates the metal cation.