Invented by Pier Adelchi Ruffini, Bjarne Bogen, Agnete Brunsvik Fredriksen, Nykode Therapeutics ASA
The Nykode Therapeutics ASA invention works as follows
The present invention relates novel recombinant fusion protein, such as human antibodies-based molecules known as Vaccibodies that are able both to trigger a B-cell- and T-cell immune response. This invention also pertains to a treatment method for cancer or infectious diseases using these specific fusion protein.Background for Homodimeric protein constructs
DNA vaccination is an easy way to induce immune responses. Clinical trials have not yet replicated the success of small animals. “Several strategies are being used to improve the efficacy and safety of DNA vaccines.
Targeting protein antigens at antigen-presenting (APC) cells can improve T and B cell responses. This can be achieved by using recombinant immunoglobulin molecules (Ig). In the Ig constant domains, for example, short epitopes may replace the loops between the?-strands. Targeted antigen delivery can be achieved by equipping recombinant Ig molecules with variable (V), specific surface molecules on APC. This strategy, however, is not suitable for antigens with unidentified epitopes or larger antigens. In addition, recombinant Ig molecule with short epitopes on T cells fail to elicit antibody against conformational epitopes. To overcome these limitations targeted Ig-based heterodimeric DNA (vaccibodies), which express infectious or tumour antigens of a minimum size of 550 aa, have been developed. “With maintenance of conformational Epitopes.
CCL3 is the human gene that encodes “Chemokine ligand (C?C motif).” CCL3, or Macrophage Inflammatory Protein-1? (MIP-1? (MIP-1? ), is a cytokine that belongs to the CC chemokine group. It is involved in acute inflammation and the recruitment of polymorphonuclear cells. The mouse CCL3 gene encodes a mature chemokine with 69 amino acid. However, the human CCL3 homolog was duplicated and mutated, leading to two non-allelic variations, LD78? The mouse CCL3 gene is a single copy and encodes a mature chemokine of 69 amino acids. However, the human homolog has been duplicated and mutated to generate two non-allelic variants: LD78 (CCL3) & LD78 (CCL3-L1) which both show 74 % homology with the mouse CCL3.” (CCL3 and LD78) show 74% homology to the mouse CCL3.
A lack of efficacy has prevented the approval of any DNA vaccine for human use. There is also no vaccine that works for many infectious diseases. “In particular, no therapeutic DNA-cancer vaccine has been approved by the FDA for human use.
WO 2004/076489 is a recombinant antibody-based molecule known as Vaccibodies that can trigger both T-cell- and B-cell immune responses.
US20070298051 is a patent that relates to using MIP-1-alpha to enhance the immune response in a mammal to an immunogen.
EP920522 is a polynucleotide vaccine containing a cDNA-target product that contains a nucleotide code encoding a specific cytokine.
Fredriksen A B et al. (Mol Ther, 2006; 13:776-85) relates to DNA vaccinations that target tumor antigen to antigen presenting cells.
Fredriksen AB and Bogen B” (Blood 2007, 110: 1797-805), relates to mouse-idiotype-fusion DNA vaccines.
OBJECT OF INVENTION
It is the object of embodiments to provide fusion protein, which can trigger an effective immune response to even weak antigens such as idiotypics antigens derived, for example, from myeloma cells. “It is an object of embodiments of the invention to provide fusion proteins, which are able to trigger an efficient immune response for even weak antigens, such as idiotypic antigens derived from e.g.
Furthermore, embodiments of the present invention aim to provide polynucleotides such as DNA polynucleotides encoding a protein fusion that triggers an efficient immune reaction against weak antigens such as idiotypics antigens derived, for example, from myeloma cells. Myeloma cell. These polynucleotides can be used to stimulate the immune system or as a vaccine against cancer or infectious diseases, which are characterized by disease-specific or disease-associated antigens.
The present inventors have found that both the full-length and truncated version of human chemokine LD78, can be used as targeting units to target antigenic epitopes on the surface APC. The chemokine or its truncated form is bound to the chemokine receptors of APC as a homodimeric construct. This allows two identical chemokines to be bound for more efficient signaling and targeting. The homodimeric protein construct also ensures that two identical epitopes of antigen are delivered to APC, which then presents them to T-cells. “Even with their relatively large size, homodimeric proteins constructs allow cells to produce and export intact molecule.
The present invention is a homodimeric amino acid protein consisting of two identical amino acids chains. Each amino acid chain contains a targeting sequence that has at least 80% sequence similarity to the amino sequence 5-70 in SEQ ID No:1, as well as an antigenic sequence.
The present invention is a homodimeric amino acid protein consisting of two identical amino acids chains. Each amino acid chain contains a targeting units comprising aminos 3-70 from SEQ ID No:1 and an antigenic, with the targeting unit and antigenic being connected by a dimerization motifs.
In a third aspect of the invention, the nucleic acid molecules encoding monomeric proteins can be converted into homodimeric proteins according to the invention.
In another aspect, the invention is a homodimeric peptide according to the invention. It can be used as a medicine.
In another aspect, the present patent relates to nucleic acids encoding monomeric proteins that can form homodimeric proteins according to the invention. For use as a drug.
The present invention also relates to a pharmaceutical formulation containing a homodimeric proteins according to the invention.
The present invention also relates to a pharmacological composition that contains a nucleic-acid molecule encoding a monomeric protein, which can be converted into a homodimeric proteins according to the invention.
The present invention also relates to host cells that contain a nucleic-acid molecule that encodes the monomeric proteins which can be converted into homodimeric proteins according to the invention.
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