Self-assembling multimeric nucleic acid constructs

• US 5,561,043 A
• Filed: 01/31/1994
• Issued: 10/01/1996
• Est. Priority Date: 01/31/1994
• Status: Expired due to Term

• Current Assignee: Trustees of Boston University, University Of Massachusetts
• Original Assignee: Trustees of Boston University
• Inventors: Sano, Takeshi, Cantor, Charles R., Rusckowski, Mary, Niemeyer, Christof M., Smith, Cassandra L., Hnatowich, Donald J.
• Primary Examiner(s): Elliott, George C.
• Assistant Examiner(s): Priebe, Scott David
• Application Number:
  US08/189,448
• Time in Patent Office: 974 Days
• Field of Search: 435/6, 536/24.3, 536/23.1
• US Class Current: 435/6.16
• CPC Class Codes: A61K 47/55 the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug, i.e. a dimer, oligomer or polymer of pharmacologically or therapeutically active compounds
• CPC Class Codes: A61K 47/557 the modifying agent being biotin
• CPC Class Codes: A61K 47/6949 inclusion complexes, e.g. clathrates, cavitates or fullerenes
• CPC Class Codes: A61K 51/1268 host-guest, closed hollow molecules, inclusion complexes, e.g. with cyclodextrins, clathrates, cavitates, fullerenes
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: C03C 17/30 with silicon-containing compounds
• CPC Class Codes: C03C 17/3405 with at least two coatings of organic materials C03C17/36, C03C17/42 take precedence
• CPC Class Codes: C12Q 1/6804 Nucleic acid analysis using immunogens immunoassay G01N33/53
• CPC Class Codes: C12Q 1/682 Signal amplification
• CPC Class Codes: Y10S 977/80 Nucleic acid, e.g. DNA or RNA
• CPC Class Codes: Y10S 977/808 Exterior attachment for targeting, e.g. drug targeting
• CPC Class Codes: Y10S 977/898 Enzymatic
• CPC Class Codes: Y10S 977/906 Drug delivery
• CPC Class Codes: Y10S 977/92 Detection of biochemical

Abstract

The invention is directed to constructs and compositions containing multimeric forms of nucleic acid. Multimeric nucleic acids comprise single-stranded nucleic acids attached via biotin to streptavidin and bound with a functional group. These constructs can be utilized in vivo to treat or identify diseased tissue or cells. Repeated administrations of multimeric nucleic acid compositions produce a rapid and specific amplification of nucleic acid constructs and their attached functional groups. For treatment purposes, functional groups may be toxins, radioisotopes, genes or enzymes. Diagnostically, labeled multimeric constructs may be used to identify specific targets in vivo or in vitro. Multimeric nucleic acids may also be used in nanotechnology and to create self-assembling polymeric aggregates such as membranes of defined porosity, microcircuits and many other products.

Claims

1. A self-assembling aggregate comprising

a construct comprising a first streptavidin or avidin molecule, having a plurality of first biotinylated single-stranded nucleic acids bound to said first streptavidin or avidin molecule;

a plurality of second biotinylated single-stranded nucleic acids bound to a second streptavidin or avidin molecule to form a multimeric nucleic acid, at least one of said second nucleic acids hybridizing with a complementary sequence of one of said first nucleic acids; and

a functional group attached to a third single-stranded nucleic acid, said third single-stranded nucleic acid hybridizing with complementary sequence of one of said second single-stranded nucleic acids bound to said multimeric nucleic acid.

2. The aggregate of claim 1 wherein the nucleic acids are selected from the group consisting of DNA, RNA and PNA (polyamide nucleic acid).

3. The aggregate of claim 1 wherein the nucleic acids are between 5 to 50 nucleotides in length.

4. The aggregate of claim 1 wherein the nucleic acids are between 8 to 25 nucleotides in length.

5. The aggregate of claim 1 wherein each of the plurality of first nucleic acids are identical.

6. The aggregate of claim 1 further comprising a biotinylated antibody bound to said first streptavidin or avidin molecule.

7. The aggregate of claim 1 wherein the functional group is selected from the group consisting of radioisotopes, pharmaceutically active moieties, genes, gene fragments, antigens, antibodies, antibody fragments, and combinations thereof.

8. The aggregate of claim 1, wherein the functional group is selected from the group consisting of toxins, cytokines metals, metabolic analogs, enzymes, oxidizing agents, bacteriostatic and bacteriocidal agents, and combinations thereof.

9. The aggregate of claim 1 wherein the functional group is selected from the group consisting of proteins, peptides, and nucleic acids, and combinations thereof.

10. A self-assembling aggregate comprising

a first construct comprising a plurality of first single-stranded nucleic acids bound to a first coupling agent forming a first multimer;

a second construct comprising a plurality of second single-stranded nucleic acids bound to a second coupling agent forming a second multimer, at least one of said second single-stranded nucleic acids hybridizing with a complementary sequence of one of said first single-stranded nucleic acids; and

a plurality of third single-stranded nucleic acids, each of said third single-stranded nucleic acids being attached to a functional group, and each of said third single-stranded nucleic acids hybridizing with a complementary sequence of a single-stranded nucleic acid attached to the aggregate,

wherein the first counting agent and the second coupling agent each include a protein, a 5'"'"'-amino-containing oligonucleotide, a 5'"'"'-thiol-containing oligonucleotide, a polyamidoamine or a divalent linker.

11. The aggregate of claim 10 wherein at least one of the first coupling agent and the second coupling agent is a protein selected from the group consisting of streptavidin, avidin, antibodies, protein A chimeras, and specific binding fragments thereof, the respective first or second single-stranded nucleic acids being conjugated to a moiety specifically bound by said protein or fragment.

12. The aggregate of claim 10, wherein the first construct and the second construct each comprise from three to seven single-stranded nucleic acids.

13. The aggregate of claim 10, wherein the plurality of third single-stranded nucleic acids do not hybridize with any nucleic acid bound to said first coupling agent.

14. The aggregate of claim 10, wherein at least one of the first coupling agent and the second coupling agent is selected from the group consisting of 5'"'"'-amino-containing oligonucleotides, 5'"'"'-thiol-containing oligonucleotides and polyamidoamines.

15. The aggregate of claim 10 wherein at least one of the first coupling agent and the second coupling agent are selected from the group consisting of streptavidin and avidin.

16. The aggregate of claim 10, wherein at least one of the first construct and the second construct is a dendrimer.