Compounds and conjugates for identifying and separating post-translationally modified analytes

Patent metadata

Field	Value
Publication number	US-2017052154-A1
Application number	US-201615239544-A
Country	US
Kind code	A1
Filing date	Aug 17, 2016
Priority date	Aug 18, 2015
Publication date	Feb 23, 2017
Grant date	—

Abstract

Disclosed herein are embodiments of compounds, conjugates, and devices, such as columns comprising such compounds and/or conjugates, that can be used to identify, separate, and quantify post-translationally modified analytes. The disclosed compounds and conjugates can be used to discriminate between analytes, such as peptides, having different post-translation modifications, such as methylations, phosphorylations, acetylations, citrullinations, hydroxylations, nitrosylations, ADP-ribosylations, glycosylations, propionylations, butyrylations, crotonylations, 2-hydroxyisobutyrylations, malonylations, succinylations, formylations, ubiquitinations, neddylations, proline cis-trans isomerizations. In particular disclosed embodiments, the compounds and conjugates can be used to separate peptides having different degrees of methylation.

First claim

We claim:

1 . A conjugate, comprising:
a compound having a structure satisfying Formula I

wherein
each of A, E, G, and J independently is —N(R b ) 2 ; halogen; —SO 3 − , —CO 2 − ; aryl; heteroaryl; -linker-aryl; or -linker-heteroaryl;
each of L, M, N, and Q independently is —OR b ; —O − ; —SH; —S − ; —N(R b ) 2 , -linker-aryl; or -linker-heteroaryl;
each of X 1 , X 2 , X 3 , and X 4 independently is CH 2 , O, S, or NR b ;
n is an integer selected from 1 to 3;
each R b independently is hydrogen, aliphatic, heteroaliphatic, aryl, or heteroaryl; and
a support component coupled to at least one of A, E, G, J, L, M, N, or Q.

2 . The conjugate of claim 1 , wherein the support component comprises a resin, a bead, a polymeric matrix, a metal oxide, a powder, a crystalline compound, an amorphous compound, or a combination thereof.

3 . The conjugate of claim 1 , wherein the support component comprises agarose, sepharose, cellulose, modified cellulose, dextran, polyacrylamide, polystyrene, latex, bonded silica gel, silica based solid, activated alumina, a polysaccharide polymer, a resinous polymer, or a combination thereof.

4 . The conjugate of claim 1 , wherein one or more of A, E, G, J, L, M, N, and Q is —N(R b ) 2 (wherein R b independently is hydrogen, aliphatic, heteroaliphatic, aryl, or heteroaryl), aryl, heteroaryl, —NR b C(O)-aryl (wherein R b independently is hydrogen, aliphatic, heteroaliphatic, aryl, or heteroaryl), —NR b C(O)-heteroaryl (wherein R b independently is hydrogen, aliphatic, heteroaliphatic, aryl, or heteroaryl), —NR b SO 2 -aryl (wherein R b independently is hydrogen, aliphatic, heteroaliphatic, aryl, or heteroaryl), —NR b SO 2 heteroaryl (wherein R b independently is hydrogen, aliphatic, heteroaliphatic, aryl, or heteroaryl), —OC(O)-aryl, —OC(O)-heteroaryl, —OC(O)O-aryl, —OC(O)O-heteroaryl, —OSO 2 -aryl, —OSO 2 -heteroaryl, —SC(O)-aryl, or —SC(O)-heteroaryl; and the remaining of A, E, G, J, L, M, N, and Q are independently —SO 3 − , —CO 2 − , —OH, or —O − .

5 . The conjugate of claim 1 , wherein aryl is phenyl or phenyl substituted with an aliphatic moiety, a heteroaliphatic moiety, a halogen, a heteroatom-containing moiety, or a combination thereof.

6 . The conjugate of claim 5 , wherein the aliphatic moiety is alkyl, alkenyl, or alkynyl; the heteroaliphatic is moiety is alkoxy, ether, thioether, amine (—NHR b , —NR b R c , or —(CH 2 ) p NHR b , wherein R b and R c independently are hydrogen, aliphatic, heteroaliphatic, aryl, or heteroaryl; and p is an integer selected from 0 to 10); the halogen is chloro, iodo, bromo, or fluoro; and the heteroatom-containing moiety is aldehyde (—(CH 2 ) p C(O)H), acyl halide (—(CH 2 ) p C(O)X, wherein X is selected from fluorine, chlorine, bromine, and iodine), carbonate (—(CH 2 ) p OC(O)OR b , wherein R b is hydrogen, aliphatic, heteroaliphatic, aryl, or heteroaryl), carboxyl (—(CH 2 ) p C(O)OH), carboxylate (—(CH 2 ) p COO − ), ester (—(CH 2 ) p C(O)OR b ), hydroxyl (—(CH 2 ) p OH), ketone (—(CH 2 ) p C(O)R b ), peroxy (—(CH 2 ) p OOR b , wherein R b is hydrogen, aliphatic, heteroaliphatic, aryl, or heteroaryl), hydroperoxy (—(CH 2 ) p OOH), phosphate (—(CH 2 ) p OP(O)OH 2 ), phosphoryl (—(CH 2 ) p P(O)(OH) 2 ), phosphodiester [—(CH 2 ) p (O) p (OH)OR b ), wherein R b is hydrogen, aliphatic, heteroaliphatic, aryl, or heteroaryl], thiol (—(CH 2 ) p SH), disulfide (—(CH 2 ) p SSR b , wherein R b is hydrogen, aliphatic, heteroaliphatic, aryl, or heteroaryl), sulfonate (—(CH 2 ) p SO 3 − ), sulfinyl (—(CH 2 ) p S(O)R b , wherein R b is hydrogen, aliphatic, heteroaliphatic, aryl, or heteroaryl), sulfonate ester (—(CH 2 ) p SO 2 OR b , wherein R b is hydrogen, aliphatic, heteroaliphatic, aryl, or heteroaryl), sulfonyl (—(CH 2 ) p SO 2 OR b , wherein R b is hydrogen, aliphatic, heteroaliphatic, aryl, or heteroaryl), carbonothioyl (—(CH 2 ) p C(S)R b or —(CH 2 ) p C(S)H, wherein R b is hydrogen, aliphatic, heteroaliphatic, aryl, or heteroaryl), sulfino (—(CH 2 ) p S(O)OH), sulfo (—(CH 2 ) p SO 3 H), thiocyanate (—(CH 2 ) p SCN), isothiocyanate (—(CH 2 ) p NCS), oxazole, oxadiazole, imidazole, triazole, tetrazole, amide (—(CH 2 ) p C(O)NR b R c , wherein R b and R c independently are hydrogen, aliphatic, heteroaliphatic, aryl, or heteroaryl), azide (—(CH 2 ) p N 3 ), azo (—(CH 2 ) p NNR b , wherein R b is hydrogen, aliphatic, heteroaliphatic, aryl, or heteroaryl), isocyanate (—(CH 2 ) p NCO), imide (—(CH 2 ) p C(O)NR b C(O)R c , wherein R b and R c independently are hydrogen, aliphatic, heteroaliphatic, aryl, or heteroaryl), nitrile (—(CH 2 ) p CN), isonitrile (—(CH 2 ) p N + ═C − ), nitro (—(CH 2 ) p NO 2 ), nitroso (—(CH 2 ) p NO), nitromethyl (—(CH 2 ) p CH 2 NO 2 ), or —(CH 2 ) p NH 2 , wherein each p independently is an integer selected from 0 to 10.

7 . The conjugate of claim 1 , wherein:
each of A, E, G, and J independently is —NH 2 , —N(C 1 -C 10 alkyl) 2 , Cl, F, Br, or I, —SO 3 − , —CO 2 − , —(CH 2 ) p Ph(CH 2 ) p (Y) m , —NHC(O)Ph(CH 2 ) p (Y) m , —NHSO 2 Ph(CH 2 ) p (Y) m , —OC(O)Ph(CH 2 ) p (Y) m , or —OSO 2 Ph(CH 2 ) p (Y) m , wherein each Y independently is positioned ortho, meta, or para on the Ph group and is aliphatic, aryl, halogen, heteroaliphatic, heteroaryl, a heteroatom-containing function group, or a combination thereof, each m is an integer selected from 0 to 4, and each p is an integer selected from 0 to 10; each L, M, N, and Q independently is —OH, —O − , —OC(O)Ph(CH 2 ) p (Y) m , —O(CH 2 ) p Y, or —OSO 2 Ph(CH 2 ) p (Y) m , wherein each Y independently is positioned ortho, meta, or para on the Ph ring and is aliphatic, aryl, halogen, heteroaliphatic, heteroaryl, a heteroatom-containing function group, or a combination thereof, and m is an integer selected from 0 to 4; each of X 1 , X 2 , X 3 , and X 4 independently can be CH 2 or S; and n is 1.

8 . The conjugate of claim 1 , wherein the compound has a formula

wherein each Y is —(CH 2 ) p C(O)NH 2 , —(CH 2 ) p C(O)OH, —(CH 2 ) p NH 2 , or —(CH 2 ) p NH 3 + , each p independently is an integer selected from 0 to 10, and each m is an integer selected from 0 to 4.

9 . The conjugate of claim 1 , wherein the compound has a formula

Assignees

• Uvic Ind Partnerships Inc

Inventors

• Hof Fraser
• Garnett Graham A E

Classifications

• C08B15/00 Primary

  Preparation of other cellulose derivatives or modified cellulose {, e.g. complexes} · CPC title

• B01J20/286

  Phases chemically bonded to a substrate, e.g. to silica or to polymers · CPC title

• C08B37/0039

  Agar; Agarose, i.e. D-galactose, 3,6-anhydro-D-galactose, methylated, sulfated, e.g. from the red algae Gelidium and Gracilaria; Agaropectin; Derivatives thereof, e.g. Sepharose, i.e. crosslinked agarose · CPC title

• G01N2030/067

  by reaction, e.g. derivatising the sample · CPC title

• C08F257/02

  on to polymers of styrene or alkyl-substituted styrenes · CPC title