Method of expanded porphyrin-oligonucleotide conjugate synthesis

• US 5,565,552 A
• Filed: 06/07/1995
• Issued: 10/15/1996
• Est. Priority Date: 01/21/1992
• Status: Expired due to Fees

• Current Assignee: Pharmacyclics Incorporated (Abbvie Incorporated), Board of Regents of the University of Texas System (University of Texas System)
• Original Assignee: Pharmacyclics Incorporated (Abbvie Incorporated), Board of Regents of the University of Texas System (University of Texas System)
• Inventors: Sessler, Jonathan L., Iverson, Brent L., Magda, Darren, Sansom, Petra I., Crofts, Shaun P.
• Primary Examiner(s): Raymond, Richard L.
• Application Number:
  US08/487,722
• Time in Patent Office: 496 Days
• Field of Search: 540/145, 540/465, 540/472, 534/11, 534/13, 534/15, 534/16, 530/345, 530/391.3, 530/391.5, 530/405, 536/17.3, 536/17.4
• US Class Current: 534/11
• CPC Class Codes: A61K 41/0038 Radiosensitizing, i.e. administration of pharmaceutical agents that enhance the effect of radiotherapy radiotherapy per se A61N5/10
• CPC Class Codes: A61K 41/0042 Photocleavage of drugs in vivo, e.g. cleavage of photolabile linkers in vivo by UV radiation for releasing the pharmacologically-active agent from the administered agent; photothrombosis or photoocclusion
• CPC Class Codes: A61K 41/0076 PDT with expanded (metallo)porphyrins, i.e. having more than 20 ring atoms, e.g. texaphyrins, sapphyrins, hexaphyrins, pentaphyrins, porphocyanines
• CPC Class Codes: A61K 47/54 the modifying agent being an organic compound
• CPC Class Codes: A61K 47/546 Porphyrines; Porphyrine with an expanded ring system, e.g. texaphyrine
• CPC Class Codes: A61K 49/0021 the fluorescent group being a small organic molecule
• CPC Class Codes: A61K 49/0052 Small organic molecules oligomers, polymers, dendrimers A61K49/0054
• CPC Class Codes: A61K 49/0054 Macromolecular compounds, i.e. oligomers, polymers, dendrimers
• CPC Class Codes: A61K 49/06 Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
• CPC Class Codes: A61K 49/085 conjugated systems
• CPC Class Codes: A61K 49/10 Organic compounds
• CPC Class Codes: A61K 49/106 the complex-forming compound being cyclic, e.g. DOTA
• CPC Class Codes: A61K 49/126 Linear polymers, e.g. dextran, inulin, PEG
• CPC Class Codes: A61K 51/0485 Porphyrins, texaphyrins wherein the nitrogen atoms forming the central ring system complex the radioactive metal
• CPC Class Codes: A61L 2/0011 using physical methods
• CPC Class Codes: B01J 2531/025 Ligands with a porphyrin ring system or analogues thereof, e.g. phthalocyanines, corroles
• CPC Class Codes: B01J 2531/26 Zinc
• CPC Class Codes: B01J 2531/27 Cadmium
• CPC Class Codes: B01J 2531/33 Indium
• CPC Class Codes: B01J 2531/37 Lanthanum
• CPC Class Codes: B01J 2531/38 : Lanthanides other than lanthanum
• CPC Class Codes: B01J 31/0201 : Oxygen-containing compounds
• CPC Class Codes: B01J 31/061 : Chiral polymers
• CPC Class Codes: B01J 31/1658 : immobilised by covalent linkages, i.e. pendant complexes with optional linking groups, e.g. on Wang or Merrifield resins
• CPC Class Codes: B01J 31/1815 : with more than one complexing nitrogen atom, e.g. bipyridyl, 2-aminopyridine
• CPC Class Codes: C07D 487/22 : in which the condensed system contains four or more hetero rings
• 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: C12N 15/113 : Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 2310/15 : Nucleic acids forming more than 2 strands, e.g. TFOs
• CPC Class Codes: C12N 2310/321 : 2'-O-R Modification
• CPC Class Codes: C12N 2310/3511 : intercalating or cleaving agent
• CPC Class Codes: C12N 2310/3521 : Methyl

Abstract

The invention is directed to a method of incorporating expanded porphyrins, and particularly of incorporating a sapphyrin or a texaphyrin, before, during, or after chemical synthesis of an oligomer to form an expanded porphyrin-oligonucleotide conjugate, and particularly a sapphyrin- or texaphyrin-oligonucleotide conjugate. This method includes reacting derivatized nucleotides and a sapphyrin or a texaphyrin in a desired order in an automated or manual DNA synthesizer having a solid support to form a sapphyrin- or a texaphyrin-oligonucleotide conjugate.

Claims

1. A method of synthesizing an expanded porphyrin-nucleotide conjugate in a DNA synthesizer, the method comprising reacting an expanded porphyrin with a nucleotide using the DNA synthesizer, to form an expanded porphyrin-nucleotide conjugate.

2. The method of claim 1 wherein the expanded porphyrin is a rubyrin.

3. The method of claim 1 wherein the expanded porphyrin is a sapphyrin.

4. The method of claim 1 wherein the expanded porphyrin is a texaphyrin.

5. The method of claim 4 wherein the texaphyrin is a phosphoramidite derivative.

6. The method of claim 1 wherein the synthesizer is an automated synthesizer.

7. The method of claim 1 wherein the nucleotide is an oligonucleotide.

8. The method of claim 7 wherein the oligonucleotide is an oligonucleotide derivative.

9. The method of claim 8 wherein the oligonucleotide derivative is selected from the group consisting of phosphorothioates, phosphoramidates, methylphosphonates and phosphotriesters.

10. The method of claim 8 wherein the oligonucleotide derivative is a 2'"'"'-O-methyl ribonucleotide.

11. A method of obtaining a 3'"'"'-linked expanded porphyrin-nucleotide conjugate, the method comprising the steps of:

attaching an expanded porphyrin to a solid support of a DNA synthesizer;

attaching a nucleic acid residue to the expanded porphyrin, followed by the attachment of one or more additional nucleic acid residues in stepwise synthesis to obtain a desired oligonucleotide;

cleaving the resulting 3'"'"'-linked expanded porphyrin-nucleotide conjugate from the solid support; and

deprotecting the oligonucleotide.

12. The method of claim 11 wherein the expanded porphyrin is a sapphyrin.

13. The method of claim 11 wherein the expanded porphyrin is a texaphyrin.

14. The method of claim 13 wherein the texaphyrin is a phosphoramidite derivative.

15. The method of claim 11 wherein the synthesizer is an automated synthesizer.

26. The method of claim 11 wherein the oligonucleotide is an oligonucleotide derivative.

27. The method of claim 11 wherein the expanded porphyrin is a phosphoramidite derivative of a texaphyrin metal complex where the texaphyrin metal complex has the following formula:

##STR6## wherein, M is H, a divalent metal cation or a trivalent metal cation;

R₁ -R₄, R₇ and R₈ are independently hydrogen, halide, hydroxyl, alkyl, aryl, haloalkyl, nitro, formyl, acyl, hydroxyalkyl, oxyalkyl, oxyhydroxyalkyl, saccharide, carboxy, carboxyalkyl, carboxyamide, carboxyamidealkyl, aminoalkyl, sulfonatoalkyl, amidealkyl, or aryl;

R₆ and R₉ are independently selected from the groups of R₁ -R₄, R₇ and R₈, with the proviso that the halide is other than iodide and the haloalkyl is other than iodoalkyl;

R₅ and R₁₀ -R₁₂ are independently hydrogen, alkyl, aryl, hydroxyalkyl, oxyalkyl, oxyhydroxyalkyl, carboxyalkyl, or carboxyamidealkyl; and

Z is an integer less than or equal to 5.

16. A method of obtaining a 5'"'"'-linked expanded porphyrin-nucleotide conjugate, the method comprising the steps of:

attaching one or more nucleic acid residues to a solid support of a DNA synthesizer in stepwise synthesis to obtain a desired oligonucleotide;

attaching an expanded porphyrin to the oligonucleotide as the final residue;

cleaving the resulting 5'"'"'-linked expanded porphyrin-nucleotide conjugate from the solid support; and

deprotecting the oligonucleotide.

17. The method of claim 16 wherein the expanded porphyrin is a sapphyrin.

18. The method of claim 16 wherein the expanded porphyrin is a texaphyrin.

19. The method of claim 18 wherein the texaphyrin is a phosphoramidite derivative.

20. The method of claim 16 wherein the synthesizer is an automated synthesizer.

28. The method of claim 16 wherein the oligonucleotide is an oligonucleotide derivative.

29. The method of claim 16 wherein the expanded porphyrin is a phosphoramidite derivative of a texaphyrin metal complex where the texaphyrin metal complex has the following formula:

##STR7## wherein, M is H, a divalent metal cation or a trivalent metal cation;

R₁ -R₄, R₇ and R₈ are independently hydrogen, halide, hydroxyl, alkyl, aryl, haloalkyl, nitro, formyl, acyl, hydroxyalkyl, oxyalkyl, oxyhydroxyalkyl, saccharide, carboxy, carboxyalkyl, carboxyamide, carboxyamidealkyl, aminoalkyl, sulfonatoalkyl, amidealkyl, or aryl;

R₆ and R₉ are independently selected from the groups of R₁ -R₄, R₇ and R₈, with the proviso that the halide is other than iodide and the haloalkyl is other than iodoalkyl;

R₅ and R₁₀ -R₁₂ are independently hydrogen, alkyl, aryl, hydroxyalkyl, oxyalkyl, oxyhydroxyalkyl, carboxyalkyl, or carboxyamidealkyl; and

Z is an integer less than or equal to 5.

21. A method of synthesizing a texaphyrin-nucleotide conjugate in a DNA synthesizer, the method comprising reacting a texaphyrin with a nucleotide using the DNA synthesizer, to form a texaphyrin-nucleotide conjugate.

22. The method of claim 21 wherein the texaphyrin is a phosphoramidite derivative of a texaphyrin metal complex where the texaphyrin metal complex has the following formula:

##STR5## wherein, M is H, a divalent metal cation or a trivalent metal cation;

R₁ -R₄, R₇ and R₈ are independently hydrogen, halide, hydroxyl, alkyl, aryl, haloalkyl, nitro, formyl, acyl, hydroxyalkyl, oxyalkyl, oxyhydroxyalkyl, saccharide, carboxy, carboxyalkyl, carboxyamide, carboxyamidealkyl, aminoalkyl, sulfonatoalkyl, amidealkyl, or aryl;

R₆ and R₉ are independently selected from the groups of R₁ -R₄, R₇ and R₈, with the proviso that the halide is other than iodide and the haloalkyl is other than iodoalkyl;

R₅ and R₁₀ -R₁₂ are independently hydrogen, alkyl, aryl, hydroxyalkyl, oxyalkyl, oxyhydroxyalkyl, carboxyalkyl, or carboxyamidealkyl; and

Z is an integer less than or equal to 5.

23. The method of claim 21 wherein the DNA synthesizer is an automated synthesizer.

24. The method of claim 21 wherein the nucleotide is an oligonucleotide.

25. The method of claim 24 wherein the oligonucleotide is an oligonucleotide derivative.