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Antigenic compositions and use of same in the targeted delivery of nucleic acids

a technology of antigen composition and nucleic acid, which is applied in the field of compositions for nucleic acid delivery, can solve the problems of few promising and difficult to find suitable delivery methods for these molecules in various applications

Inactive Publication Date: 2014-07-03
AKSHAYA BIO INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The patent describes a method for inhibiting the expression of a target gene in a target cell using a nucleic acid molecule and a chimeric antigen. The nucleic acid molecule is designed to target the gene and the chimeric antigen contains a nucleic acid interaction domain and a target binding domain that binds to a receptor on the surface of the target cell. The chimeric antigen can be administered to the target cell along with the nucleic acid molecule, resulting in a more effective inhibition of gene expression. The method can be used to treat mammalian hosts with a targeted antigen.

Problems solved by technology

Despite recent advances in the identification and refinement of nucleic acid therapeutics, finding suitable delivery means for these molecules in various applications has proved challenging.
Moreover, while it is desirable to minimize the dosage of these expensive molecules, by localizing or targeting nucleic acid therapies to tissues / cells of interest, many technologies have been investigated, with few promising results.
Targeted delivery of siRNA into specific cells of interest has been the main obstacle to achieving in vivo gene silencing by RNAi technologies.

Method used

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  • Antigenic compositions and use of same in the targeted delivery of nucleic acids
  • Antigenic compositions and use of same in the targeted delivery of nucleic acids
  • Antigenic compositions and use of same in the targeted delivery of nucleic acids

Examples

Experimental program
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example 1

Cloning and Expression of a Chimeric Antigen with a C-Terminal Protamine Tail

[0108]Step 1. Cloning—DNA encoding a target binding domain (TBD) containing a 5′ Not I site and a 3′ Xba I site was produced by PCR using previously generated pFastBacHTa-TBD as template with unique primers that add the respective restriction enzyme sites. The primers used were;

5′ Primer(SEQ ID NO: 8)5′ TGTCATTCTGCGGCCGCAAGGCGGCGGGATCCGTGGACAAGAAAATTGTGCCCAGG 3′3′ Primer(SEQ ID NO: 9)5′ CCGGTCTAGATTCAGCCCAGGAGAGTGGGAGAG 3′.

The PCR fragment was isolated, digested with Not I and Xba I and cloned into a Not I / Xba I digested pFastBacHTa-gp64 plasmid.

[0109]For HBV Core protamine tail, the sequence was obtained by PCR of previously produced plasmid pFastBacHTa HBV Core-TBD as template using primers that add a unique Xba I site to the 5′ end and a unique Hind III site to the 3′ end. The primers used were:

5′ Primer:(SEQ ID NO: 10)5′ CCGGTCTAGAGGAAACTACTGTTGTTAGACGAC 3′and3′ Primer:(SEQ ID NO: 11)5′ GCGCAAGCTTTGACA...

example 2

Visualization of Chimigen Aggregation

[0121]Chimigen® HBV Core Vaccine and Chimigen® HBV S1 / S2 Core Vaccine were visualized using Tapping Mode Atomic Force Microscopy (TM-AFM). The images generated are shown in FIG. 6a-d and 7a-d, respectively. As indicated, the aggregates / nanoparticles formed are of uniform size and ellipsoid shape, having a diameter of 30-40 nm and a height of 2 nm.

example 3

Encapsulation of shRNA Plasmid by Chimigen® S1 / S2 Core Vaccine

[0122]SureSilencing shRNA plasmid was mixed with Chimigen HBV S1 / S2 Core Vaccine under denaturing conditions. After removal of the denaturing conditions, encapsulation was evaluated by DNase treatment and PCR amplification of GFP DNA. The shRNA vector plasmid and results are shown in FIGS. 8a and 8b, respectively. It is noted that both vaccines protected the GFP DNA from DNAse treatment, suggesting that the vaccines are capable of forming encapsulated delivery vehicles around nucleic acids.

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Abstract

Methods and compositions are provided for delivery of therapeutic nucleic acids to a target cell. A chimeric antigen is provided to encapsulate, bind, or otherwise carry a nucleic acid molecule to a target cell where the chimeric antigen and nucleic acid are internalized, for example by receptor-mediated endocytosis. The chimeric antigen has a nucleic acid interaction domain, a target binding domain, and an immune response domain that may include a target antigen. Targeting is generally provided by the specificity of the target binding domain for a particular target cell receptor, but may also be provided by inclusion of a targeting antigen within the immune response domain. The combined delivery of chimeric antigen and nucleic acid, which may be a siRNA, may be synergistic in certain applications, for example in breaking host tolerance to a virus or in providing immunostimulation.

Description

CROSS REFERENCE TO RELATED APPLICATIONS[0001]This application is a divisional application of U.S. patent application Ser. No. 12 / 675,560, filed Jan. 28, 2011, which is national stage application under 35 U.S.C. 371 of PCT Application No. PCT / CA2008 / 01547, having an international filing date of Aug. 29, 2008, which designated the United States, which PCT application claims the benefit of priority under 35 U.S.C. §119(e) from U.S. Provisional Application No. 60 / 968,978, filed Aug. 30, 2007. The entire disclosure of each is incorporated herein by reference.FIELD OF THE INVENTION[0002]The present invention relates to compositions for use in the delivery of nucleic acids. More particularly, chimeric antigens are provided for encapsulating, binding, or otherwise carrying and delivering nucleic acids to a target cell.BACKGROUND OF THE INVENTION[0003]Despite recent advances in the identification and refinement of nucleic acid therapeutics, finding suitable delivery means for these molecules...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C07K16/28C07K14/005A61K39/00C12N15/11C12N15/87
CPCC07K14/005C07K16/28A61K39/292A61K48/00A61K48/0025A61K2039/6025A61K2039/6056A61K2039/6075A61K2039/64C07K2319/035C07K2319/21C07K2319/30C07K2319/33C07K2319/74C07K2319/80C07K2319/85C12N15/111C12N15/62C12N15/87C12N2310/14C12N2320/32C12N2730/10122C12N2730/10134C12N2810/859A61K2039/5154A61K2039/6006A61K2039/625A61K39/12A61K47/646A61P31/12A61P33/00A61P35/00A61P37/04A61P37/06
Inventor GEORGE, RAJANNOUJAIM, ANTOINE
Owner AKSHAYA BIO INC
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