Polymeric carrier cargo complex for use as an immunostimulating agent or as an adjuvant
The present invention is directed to a polymeric carrier cargo complex, comprising as a cargo at least one nucleic acid molecule and as a preferably non-toxic and non-immunogenic polymeric carrier disulfide-crosslinked cationic components for use as an immunostimulating agent or as an adjuvant, wherein the polymeric carrier cargo complex is administered in combination with at least one second nucleic acid molecule, which encodes a protein or peptide. The inventive polymeric carrier cargo complex administered in combination with the second nucleic acid molecule allows for both efficient transfection of nucleic acids into cells in vivo and in vitro and/or for induction of an innate and/or adaptive immune response, preferably dependent on the nucleic acid to be transported as a cargo and on the second nucleic acid molecule. The present invention also provides pharmaceutical compositions, particularly vaccines, comprising the inventive polymeric carrier cargo complex and the second nucleic acid molecule, as well as the use of the inventive polymeric carrier cargo complex and the second nucleic acid molecule for transfecting a cell, a tissue or an organism, as a medicament, for therapeutic purposes as disclosed herein, and/or as an immunostimulating agent or adjuvant, e.g. for eliciting an immune response for the treatment or prophylaxis of diseases as mentioned herein. Finally, the invention relates to kits containing the inventive polymeric carrier cargo complex and the second nucleic acid molecule, the inventive pharmaceutical composition and/or the inventive vaccine or any of its components in one or more parts of the kit.
1. A method of stimulating an immune response in a subject comprising administering to the subject a polymeric carrier cargo complex, comprising:
a) as a carrier a polymeric carrier formed by disulfide-crosslinked cationic components, and
b) as a cargo at least one first nucleic acid molecule,
wherein the polymeric carrier cargo complex is administered in combination with at least one second nucleic acid molecule encoding a protein or a peptide, and
wherein the polymeric carrier cargo complex and the second nucleic acid molecule are administered intramuscularly, thereby stimulating an immune response in the subject;
wherein the second nucleic acid molecule is an mRNA molecule that comprises a 5′-UTR sequence derived from a 5′-UTR of a TOP gene.
2. The method of claim 1 , wherein the second nucleic acid molecule is complexed with a cationic component.
3. The method of claim 1 , wherein the second nucleic acid molecule is not packaged in a particle, such as a virus particle, an inactivated virus particle or a virus-like particle.
4. The method of claim 1 , wherein the second nucleic acid molecule is not comprised in the polymeric carrier cargo complex.
5. The method of claim 1 , wherein the polymeric carrier cargo complex and the second nucleic acid molecule are not administered together with a protein or peptide antigen.
6. The method of claim 1 , wherein the second nucleic acid molecule is an mRNA molecule that further comprises a 3′-UTR, a poly(A) sequence, a poly(C) sequence and a histone stem-loop sequence.
7. The method of claim 1 , wherein the mRNA molecule comprises at least one selected from the group consisting of a 3′-UTR, a poly(A) sequence, a poly(C) sequence and a histone stem-loop sequence.
8. The method of claim 7 , wherein the 3′-UTR comprises a nucleic acid sequence derived from the 3′-UTR of an albumin gene.
9. The method of claim 8 , wherein the 3′-UTR comprises the nucleic acid sequence corresponding to SEQ ID NO. 388.
10. The method of claim 7 , wherein the histone stem-loop sequence comprises a nucleic acid sequence corresponding to SEQ ID NO. 389.
11. The method of claim 7 , wherein the mRNA molecule comprises a nucleic acid sequence derived from a 5′-TOP-UTR, a GC-optimized coding sequence, a nucleic acid sequence derived from the 3′-UTR of an albumin gene, a poly(A)-sequence, a poly(C)-sequence, and a histone stem loop.
12. The method of claim 1 , wherein the 5′-UTR comprises a nucleic acid sequence derived from a ribosomal protein gene.
13. The method of claim 12 , wherein the 5′-UTR comprises a nucleic acid sequence derived from ribosomal protein 32L gene.
14. The method of claim 1 , wherein the at least one first nucleic acid molecule is an RNA molecule.
15. The method of claim 1 , wherein the at least one first nucleic acid molecule is an immunostimulatory nucleic acid.
16. The method of claim 15 , wherein the immunostimulatory nucleic acid is a non-coding immunostimulatory nucleic acid.
17. The method of claim 15 , wherein the immunostimulatory nucleic acid is a RNA.