Alkynes and methods of reacting alkynes with 1,3-dipole-functional compounds

• US 8,133,515 B2
• Filed: 11/21/2008
• Issued: 03/13/2012
• 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: Boons, Geert-Jan, Guo, Jun, Ning, Xinghai, Wolfert, Margaretha
• Primary Examiner(s): LOEWE, SUN JAE Y
• Application Number:
  US12/743,632
• Publication Number:
  US 20100297250A1
• Time in Patent Office: 1,208 Days
• Field of Search: None
• 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:

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

3. 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.

4. The alkyne of claim 3 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.

5. The alkyne of claim 4 wherein the cleavable linker, if present, is a disulfide.

6. 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.

7. The alkyne of claim 6 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.

21. 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.

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

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

providing a substrate according claim 22;

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

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

providing a substrate according to claim 22;

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

8. An alkyne of the formula:

9. The alkyne of claim 8 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.

10. The alkyne of claim 9 wherein the cleavable linker, if present, is a disulfide.

11. An alkyne of the formula:

12. The alkyne of claim 11 wherein the copolymeric group comprises a hydrophilic segment and a hydrophobic segment.

13. The alkyne of claim 11 wherein the copolymeric group comprises a fragment of the formula —

[CH₂CH₂O]_n[C(O)(CH₂)₅O]_m—

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

18. A composition comprising a blend of:

an alkyne according to claim 11; and

a polymer or a copolymer.

19. The composition of claim 18 wherein the composition forms a copolymer micelle.

20. 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 19 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.

14. An alkyne of the formula:

15. An alkyne of the formula:

16. An alkyne of the formula:

17. An alkyne of the formula: