US 12,474,350 B2
Low energy cleavable mass tag for quantitative proteomics
Felix Meissner, Berlin (DE); Matthias Mann, Munich (DE); Florian Meier, Pullach (DE); Sebastian Virreira Winter, Munich (DE); and Arturo Zychlinsky, Berlin (DE)
Appl. No. 16/349,363
Filed by Bruker Switzerland AG, Faellanden (CH)
PCT Filed Nov. 14, 2017, PCT No. PCT/EP2017/079211
§ 371(c)(1), (2) Date May 13, 2019,
PCT Pub. No. WO2018/087397, PCT Pub. Date May 17, 2018.
Claims priority of application No. 16198748 (EP), filed on Nov. 14, 2016.
Prior Publication US 2019/0369112 A1, Dec. 5, 2019
This patent is subject to a terminal disclaimer.
Int. Cl. C07C 317/04 (2006.01); C07D 211/94 (2006.01); G01N 33/68 (2006.01)
CPC G01N 33/6848 (2013.01) [C07C 317/04 (2013.01); C07D 211/94 (2013.01)] 5 Claims
 
1. A compound which comprises or consists of
(a) a reactive moiety (a), said reactive moiety (a) being capable of reacting with a functional group of a peptide to form a covalent bond, and covalently joined thereto; and
(b) a moiety (b) which fragments in the mass spectrometer
(i) at an energy below the energy required for fragmenting peptides and/or a higher conversion rate than peptides; and
(ii) at said energy according to (i), and when coupled to a peptide via said reactive moiety (a), fragments at a single site within said compound coupled to said peptide, to yield a first moiety and a second moiety, said second moiety being coupled to said peptide;
wherein said compound has formula (V a) or (V b):

OG Complex Work Unit Chemistry
wherein R2 is hydrogen or an electron-withdrawing group selected from halogen, NH3+, NR3R4R5+, or NO2, and wherein R3, R4 and R5 are independently C1 to C6 alkyl or cycloalkyl, C2 to C6 alkenyl or cycloalkenyl or C2 to C6 alkinyl;
J is a moiety containing C and optionally N and/or O, optionally wherein one or more of said C, N, or O is isotope labelled; wherein said moiety J is selected from substituted or unsubstituted C1 to C6 alkyl or cycloalkyl, substituted or unsubstituted C2 to C6 alkenyl or cycloalkenyl, or substituted or unsubstituted C2 to C6 alkinyl, wherein 1 or 2 carbon atoms may be replaced with heteroatoms selected from O, N, S and P, substituents including OH, halogen, methyl and methoxy, and optionally H3C—(CH2)r—O;
n is an integer number between 0 and 19;
r is an integer number between 0 and 20; or

OG Complex Work Unit Chemistry
wherein
R2 is an electron-withdrawing group selected from halogen, NH3+, NR3R4R5+, or NO2, wherein R3, R4 and R5 are independently C1 to C6 alkyl or cycloalkyl, C2 to C6 alkenyl or cycloalkenyl or C2 to C6 alkinyl; and
r and t independently are integer numbers between 1 and 20.