ALKYNES AND METHODS OF REACTING ALKYNES WITH 1,3-DIPOLE-FUNCTIONAL COMPOUNDS

• US 20100297250A1
• Filed: 11/21/2008
• Published: 11/25/2010
• Est. Priority Date: 11/21/2007
• Status: Active Grant

• Current Assignee: The University of Georgia Research Foundation, Inc.
• Original Assignee: The University of Georgia Research Foundation, Inc.
• Inventors: Ning, Xinghai, Wolfert, Margaretha, Guo, Jun, Boons, Geert-Jan
• Granted Patent: US 8,133,515 B2
• Time in Patent Office: Days
• US Class Current: 424/501
• CPC Class Codes: A61K 47/54 the modifying agent being an organic compound
• CPC Class Codes: A61K 47/542 Carboxylic acids, e.g. a fatty acid or an amino acid
• CPC Class Codes: A61K 47/61 the organic macromolecular compound being a polysaccharide or a derivative thereof
• CPC Class Codes: A61K 47/62 the modifying agent being a protein, peptide or polyamino acid
• CPC Class Codes: A61K 47/64 Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent peptidic linkers A61K47/65
• CPC Class Codes: A61K 47/6803 Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
• CPC Class Codes: A61K 47/6921 the form being a particulate, a powder, an adsorbate, a bead or a sphere
• CPC Class Codes: A61K 49/0021 the fluorescent group being a small organic molecule
• CPC Class Codes: A61K 49/0043 Fluorescein, used in vivo
• CPC Class Codes: A61K 49/0052 Small organic molecules oligomers, polymers, dendrimers A61K49/0054
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: C07C 211/42 with six-membered aromatic rings being part of the condensed ring systems
• CPC Class Codes: C07C 251/44 with the carbon atom of at least one of the oxyimino groups being part of a ring other than a six-membered aromatic ring
• CPC Class Codes: C07C 251/58 of hydrocarbon radicals substituted by nitrogen atoms not being part of nitro or nitroso groups
• CPC Class Codes: C07C 2603/36 containing eight-membered rings
• CPC Class Codes: C07C 271/34 with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms
• CPC Class Codes: C07C 35/37 with a hydroxy group on a condensed system having three rings
• CPC Class Codes: C07C 49/683 having unsaturation outside the aromatic rings
• CPC Class Codes: C07D 249/16 condensed with carbocyclic rings or ring systems
• CPC Class Codes: C07D 261/20 condensed with carbocyclic rings or ring systems
• CPC Class Codes: C07D 271/12 : condensed with carbocyclic rings or ring systems
• CPC Class Codes: C07D 307/94 : spiro-condensed with carbocyclic rings or ring systems, e.g. griseofulvins
• CPC Class Codes: C07D 311/90 : with hydrocarbon radicals, substituted by amino radicals, directly attached in position 9
• CPC Class Codes: C07D 495/04 : Ortho-condensed systems
• CPC Class Codes: C07H 21/00 : Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C07K 14/00 : Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
• CPC Class Codes: C07K 2/00 : Peptides of undefined number of amino acids; Derivatives thereof
• CPC Class Codes: G01N 33/582 : with fluorescent label

Abstract

1,3-Dipole-functional compounds (e.g., azide functional compounds) can be reacted with certain alkynes in a cyclization reaction to form heterocyclic compounds. Useful alkynes (e.g., strained, cyclic alkynes) and methods of making such alkynes are also disclosed. The reaction of 1,3-dipole-functional compounds with alkynes can be used for a wide variety of applications including the immobilization of biomolecules on a substrate.

Claims

1. An alkyne of the formula:

4. The alkyne claim 1 wherein R³ comprises a covalently bound organic dye.

6. The alkyne of claim 1 wherein X represents C═

N—

OR³ and R³ represents an organic group having the formula —

(CH₂)_aC(O)Y, wherein;

a is 1-3;

Y represents OH or NHR⁵; and

R⁵ represents hydrogen or a biotinylation product of a primary amine-containing organic group.

7. The alkyne of claim 6 wherein the biotinylation product is the biotinylation product of a primary amine-containing group of the formula —

(CH₂CH₂O)_b(CH₂)_c-L_d (CH₂CH₂O)_e(CH₂)_fNH₂ and/or —

(CD₂CD₂O)_b-L_d-(CD₂CD₂O)_e(CD₂)_fNH₂, wherein b=0 to 100;

c=0 to 100;

d=0 to 100;

e=0 to 100;

f=0 to 100; and

L is an optional cleavable linker.

8. The alkyne of claim 7 wherein the cleavable linker, if present, is a disulfide.

9. The alkyne of claim 1 wherein X represents CHOR³ and R³ is selected from the group consisting of an alkyl group, an aryl group, an alkaryl group, and an aralkyl group.

10. The alkyne of claim 9 wherein R³ represents —

C(O)Z, wherein;

Z represents an alkyl group, OR⁶, or NHR⁷;

R⁶ and R⁷ are each independently selected from the group consisting of an alkyl group, an aryl group, an alkaryl group, and an aralkyl group.

11. The alkyne of claim 10 wherein R⁷ is a biotinylation product of a primary amine-containing organic group.

12. The alkyne of claim 11 wherein the biotinylation product is the biotinylation product of a primary amine-containing group of the formula —

(CH₂CH₂O)_b(CH₂)_c-L_d- (CH₂CH₂O)_e(CH₂)_fNH₂ and/or —

(CD₂CD₂O)_b(CD₂)_c-L_d-(CD₂CD₂O)_e(CD₂)_fNH₂, wherein b=0 to 100;

c=0 to 100;

d=0 to 100;

e=0 to 100;

f=0 to 100; and

L is an optional cleavable linker.

13. The alkyne of claim 12 wherein the cleavable linker, if present, is a disulfide.

14. The alkyne claim 1 wherein R³ represents a polymeric or a copolymeric group.

15. The alkyne of claim 14 wherein the copolymeric group comprises a hydrophilic segment and a hydrophobic segment.

16. The alkyne of claim 14 wherein the copolymeric group comprises a fragment of the formula —

[CH₂CH₂O]_n—

[C(O)(CH₂)]_m—

H, wherein n=0 to 100 and m=0 to 100.

21. A composition comprising a blend of:

an alkyne according to claim 14; and

a polymer or a copolymer.

25. The composition of claim 21 wherein the composition forms a copolymer micelle.

26. A method for controlling the delivery of drugs, the method comprising:

combining at least one 1,3-dipole-functional drug with a copolymer micelle according to claim 25 comprising an alkyne; and

allowing the at least one 1,3-dipole-functional drug and the copolymer micelle comprising the alkyne to react under conditions effective to form a heterocyclic compound.

35. A method of preparing a heterocyclic compound, the method comprising:

combining at least one 1,3-dipole-functional compound with at least one alkyne according to claim 1; and

allowing the at least one 1,3-dipole-functional compound and the at least one alkyne to react under conditions effective to form the heterocyclic compound.

60. A substrate having on the surface thereof an alkyne according to claim 1.

64. A method of immobilizing a biomolecule on a substrate, the method comprising:

providing a substrate according claim 60;

contacting the substrate with a 1,3-dipole-functional biomolecule under conditions effective to form a heterocyclic compound.

69. A method for immobilizing a cell, the method comprising:

providing a substrate according to claim 60;

contacting the substrate with a cell comprising a 1,3-dipole-functional biomolecule under conditions effective to form a heterocyclic compound.

2-3. -3. (canceled)

5. (canceled)

17. An alkyne of the formula:

18. An alkyne of the formula:

19. An alkyne of the formula:

20. An alkyne of the formula:

22-24. -24. (canceled)

27. A compound of the formula:

28. A compound of the formula:

29-30. -30. (canceled)

31. A compound of the formula:

32. A compound of the formula:

33-34. -34. (canceled)

36-59. -59. (canceled)

61-63. -63. (canceled)

65-68. -68. (canceled)