Zwitterionic reagents

Patent metadata

Field	Value
Publication number	US-9487480-B2
Application number	US-201113699435-A
Country	US
Kind code	B2
Filing date	May 18, 2011
Priority date	May 21, 2010
Publication date	Nov 8, 2016
Grant date	Nov 8, 2016

Abstract

Zwitterion-containing compounds for the modification of hydrophobic molecules to improve their solubility and/or to lower their non-specific binding as provided. The zwitterion-containing compounds may be suitable for modification of detectable labels such as biotin and fluorescein to improve their solubility. The zwitterion-containing compounds may also be useful for the preparation of conjugates of proteins, peptides and other macromolecules or for crosslinking molecules and/or macromolecules.

First claim

What is claimed:

1. A zwitterion-containing compound for forming a conjugate with a peptide, protein, or macromolecule, said compound having the structure of formula (I):

wherein,
L 1 is a divalent C 1-20 hydrocarbon radical selected from the group consisting of alkyl, alkenyl, alkynyl, aryl, and aralkyl groups, each optionally substituted with up to 20 heteroatoms independently selected from the group consisting of halogen, nitrogen, oxygen, sulfur, and phosphorus;
L 2 is a bond or a divalent C 1-4 hydrocarbon radical selected from the group consisting of alkyl, alkenyl, and alkynyl groups, each optionally substituted with up to 10 heteroatoms independently selected from the group consisting of halogen, nitrogen, oxygen, sulfur, and phosphorus:
Z is an anion-containing group selected from the group consisting of sulfonate (—SO 3 − ), sulfate (—OSO 3 − ), and phosphate (—OP(O)(OR)(O − )), where R is a C 1-12 hydrocarbon radical selected from the group consisting of alkyl, alkenyl, alkynyl, aryl, and aralkyl groups, each optionally substituted with up to 20 heteroatoms independently selected from the groups consisting of halogen, nitrogen, oxygen, sulfur, and phosphorus;
Y 1 is a reactive-functional group for forming covalent linkages with a peptide, a protein, or a macromolecule, said functional group comprising a electrophilic group, nucleophilic group, or a photoreactive group; wherein Y 1 is selected from the group consisting of:

where R is an alkyl, alkenyl, alkynyl, aryl, or aralkyl group, optionally substituted with up to 20 heteroatoms each independently selected from the group consisting of halogen, nitrogen, oxygen, sulfur, and phosphorus, and
R 1 and R 2 are independently selected at each occurrence from a C 1-20 hydrocarbon radical selected from the group consisting of alkyl, alkenyl, alkynl, aryl and aralkyl groups, each optionally substituted with up to 20 heteroatoms independently selected from the group consisting of halogen, nitrogen, oxygen, sulfur, and phosphorus.

2. The zwitterion-containing compound according to claim 1 , wherein Y 1 is N-succinimidyloxycarbonyl and Z is sulfonate.

3. The zwitterion-containing compound according to claim 1 , wherein Y 1 is selected from a pentafluorophenyl (PFP) ester, maleimide, or N-succinimidyloxycarbonyl.

4. The zwitterion-containing compound according to claim 1 , wherein L 1 is a divalent C 1-10 alkyl, optionally substituted with up to 20 heteroatoms independently selected from the group consisting of halogen, nitrogen, oxygen, sulfur, and phosphorus.

5. The zwitterion-containing compound according to claim 4 , wherein L 1 is a divalent radical of the form —(CH 2 ) n — where n=1 to 6.

6. The zwitterion-containing compound according to claim 4 , wherein L 2 is a divalent C 1-4 alkyl, optionally substituted with up to 10 heteroatoms independently selected from the group consisting of halogen, nitrogen, oxygen, sulfur, and phosphorus.

7. The zwitterion-containing compound according to claim 6 , wherein L 2 is a divalent radical of the form —(CH 2 ) m — where m=1 to 4.

8. The zwitterion-containing compound according to claim 7 , wherein Z is sulfonate (—SO 3 − ).

9. The zwitterion-containing compound according to claim 7 , wherein Z is phosphate (—OP(O)(OR)(O − )) where R is a C 1-12 hydrocarbon radical selected from the group consisting of alkyl, alkenyl, alkynyl, aryl, and aralkyl groups, each optionally substituted with up to 20 heteroatoms independently selected from the group consisting of halogen, nitrogen, oxygen, sulfur, and phosphorus.

10. The zwitterion-containing compound according to claim 7 , wherein Z is carboxylate (—COO − ).

11. The zwitterion-containing compound according to claim 8 , having the structure:

12. The zwitterion-containing compound according to claim 8 , having the structure:

13. The zwitterion-containing compound according to claim 8 , having the structure:

14. The zwitterion-containing compound according to claim 1 , having the structure:

15. A method of improving the aqueous solubility of a peptide, protein, or macromolecule comprising covalently attaching the zwitterion-containing compound according to claim 1 to said peptide, protein or macromolecule through reactive group Y 1 wherein, the zwitterion-containing compound improves the aqueous solubility of said peptide, protein, or macromolecule.

16. A method of reducing non-specific binding of an analyte, analyte analog, or binding partner for an analyte in a biologic binding assay involving a solid phase comprising covalently attaching the zwitterion-containing compound according to claim 1 to said analyte, analyte analog, or binding partner for an analyte through reactive group Y 1 wherein, non-specific binding interactions of said analyte, analyte analog, or binding partner with the solid phase are reduced due to the zwitterion-containing compound.

17. The zwitterion-containing compound according to claim 1 wherein L 1 is a divalent C 1-20 hydrocarbon radical selected from the group consisting of alkyl group, polyethylene glycol chain, and a linear moiety of the form —(CH 2 ) b O(CH 2 ) c —, —(CH 2 ) b S(CH 2 ) c —, or —(CH 2 ) b NR L (CH 2 ) c — wherein “b” and “c” are independently an integer from 1 to 18 and R L is an alkyl, alkenyl, alkynyl, aryl, or aralkyl group, each optionally substituted with up to 20 heteroatoms independently selected from the group consisting of halogen, nitrogen, oxygen, sulfur, and phosphorus.

18. The zwitterion-containing compound according to claim 17 , wherein Y 1 is selected from a pentafluorophenyl (PFP) ester, maleimide, or N-succinimidyloxycarbonyl.

19. The zwitterion-containing compound according to claim 17 , wherein Y 1 is sulfonate (—SO 3 − ).

20. The zwitterion-containing compound according to claim 17 , wherein Z is phosphate (—OP(O)(OR)(O − )) where R is a C 1-12 hydrocarbon radical selected from the group consisting of alkyl, alkenyl, alkynyl, aryl, and aralkyl groups, each optionally substituted with up to 20 heteroatoms independently selected from the group consisting of halogen, nitrogen, oxygen, sulfur, and phosphorus.

21. A zwitterion-containing compound for forming a conjugate with a peptide, protein, or macromolecule, said compound having the structure of formula (I):

wherein,
L 1 is a divalent C 1-20 hydrocarbon radical selected from the group consisting of alkyl, alkenyl, alkynyl, aryl, and aralkyl groups, each optionally substituted with up to 20 heteroatoms independently selected from the group consisting of halogen, nitrogen, oxygen, sulfur, and phosphorus;
L 2 is a bond or a divalent C 1-4 hyd

Assignees

• Natrajan Anand
• Sharpe David
• Wen David
• Jiang Qingping
• Siemens Healthcare Diagnostics Inc

Inventors

• Natrajan Anand
• Sharpe David
• Wen David
• Jiang Qingping

Classifications

• A61K47/48061

  Human Necessities · mapped topic

• C07D403/12

  linked by a chain containing hetero atoms as chain links · CPC title

• C07F9/091

  with hydroxyalkyl compounds with further substituents on alkyl · CPC title

• A61K47/48284

  Human Necessities · mapped topic

• C07C309/14

  containing amino groups bound to the carbon skeleton · CPC title