Polynucleotides encoding anti-chikungunya virus antibodies
This disclosure relates to compositions and methods for treating and preventing chikungunya virus infection by delivering polynucleotides encoding anti-chikungunya virus antibodies to a subject. Compositions and treatments provided herein include one or more polynucleotides having an open reading frame encoding an anti-chikungunya virus antibody heavy chain or fragment thereof and/or an anti-chikungunya virus antibody light chain or fragment thereof. Methods for preparing and using such treatments are also provided.
1. A polynucleotide comprising an mRNA comprising:
(i) a 5′ UTR;
(ii) an open reading frame (ORF) encoding a polypeptide comprising the heavy chain variable region of the heavy chain antibody sequence of SEQ ID NO:1, wherein the ORF comprises a nucleic acid sequence that is at least 80% identical to nucleotides 61-426 of SEQ ID NO:2;
(iii) a stop codon; and
(iv) a 3′ UTR.
2. A polynucleotide comprising an mRNA comprising:
(i) a 5′ UTR;
(ii) an open reading frame (ORF) encoding a polypeptide comprising the light chain variable region of the light chain antibody sequence of SEQ ID NO:3, wherein the ORF comprises a nucleic acid sequence that is at least 80% identical to nucleotides 61-384 of SEQ ID NO:4;
(iii) a stop codon; and
(iv) a 3′ UTR.
3. A pharmaceutical composition comprising the polynucleotide of claim 1 , and a delivery agent.
4. A pharmaceutical composition comprising:
a first polynucleotide comprising a first mRNA comprising (i) a first 5′ UTR, (ii) a first open reading frame (ORF) encoding a first polypeptide comprising the heavy chain variable region of the heavy chain antibody sequence of SEQ ID NO:1, wherein the first ORF comprises a first nucleic acid sequence that is at least 80% identical to nucleotides 61-426 of SEQ ID NO:2, (iii) a first stop codon, and (iv) a first 3′ UTR;
a second polynucleotide comprising a second mRNA comprising (i) a second 5′ UTR, (ii) a second ORF encoding a second polypeptide comprising the light chain variable region of the light chain antibody sequence of SEQ ID NO:3, wherein the second ORF comprises a second nucleic acid sequence that is at least 80% identical to nucleotides 61-384 of SEQ ID NO:4, (iii) a second stop codon, and (iv) a second 3′ UTR; and
a delivery agent,
wherein the first polypeptide when paired with the second polypeptide forms an anti-Chikungunya virus antibody or an anti-Chikungunya virus antibody fragment.
5. The pharmaceutical composition of claim 4 , wherein the first nucleic acid sequence is at least 90% identical to SEQ ID NO:2, and wherein the second nucleic acid sequence is at least 90% identical to SEQ ID NO:4.
6. The pharmaceutical composition of claim 4 , wherein the first mRNA and the second mRNA each comprise a 5′ terminal cap.
7. The pharmaceutical composition of claim 6 , wherein each 5′ terminal cap comprises a Cap0, Cap0, ARCA, inosine, N1-methyl-guanosine, 2′-fluoro-guanosine, 7-deaza-guanosine, 8-oxo-guanosine, 2-amino-guanosine, LNA-guanosine, 2-azidoguanosine, Cap2, Cap4, 5′ methylG cap, or an analog thereof.
8. The pharmaceutical composition of claim 4 , wherein the first mRNA and the second mRNA each comprise a poly-A region.
9. The pharmaceutical composition of claim 4 , wherein the first mRNA and the second mRNA each comprise at least one chemically modified nucleobase, sugar, backbone, or any combination thereof.
10. The pharmaceutical composition of claim 9 wherein the at least one chemically modified nucleobase is selected from the group consisting of pseudouracil (ψ), N1-methylpseudouracil (m1ψ), 1-ethylpseudouracil, 2-thiouracil (s2U), 4′-thiouracil, 5-methylcytosine, 5-methyluracil, 5-methoxyuracil, and any combination thereof.
11. The pharmaceutical composition of claim 9 , wherein all of the uracils of the first mRNA and the second mRNA are N1-methylpseudouracils.
12. A method of expressing an anti-chikungunya virus antibody in a human subject in need thereof, comprising administering to the human subject an effective amount of the pharmaceutical composition of claim 4 .
13. A method of reducing chikungunya virus levels in a human subject in need thereof, comprising administering to the human subject an effective amount of the pharmaceutical composition of claim 4 .
14. The method of claim 12 , wherein the pharmaceutical composition is administered to the human subject multiple times at a frequency of about once a week, about once every two weeks, or about once a month.
15. The method of claim 12 , wherein the pharmaceutical composition is administered intravenously.
16. The method of claim 12 , wherein the pharmaceutical composition is administered subcutaneously.
17. The pharmaceutical composition of claim 4 , wherein the first nucleic acid sequence is at least 80% identical to nucleotides 61-1416 of SEQ ID NO:2, and wherein the second nucleic acid sequence is at least 80% identical to nucleotides 61-705 of SEQ ID NO:4.
18. The pharmaceutical composition of claim 4 , wherein the first nucleic acid sequence is at least 80% identical to SEQ ID NO:2, and wherein the second nucleic acid sequence is at least 80% identical to SEQ ID NO:4.
19. The pharmaceutical composition of claim 4 , wherein the first nucleic acid sequence is at least 90% identical to nucleotides 61-426 of SEQ ID NO:2, and wherein the second nucleic acid sequence is at least 90% identical to nucleotides 61-384 of SEQ ID NO:4.
20. The pharmaceutical composition of claim 4 , wherein the first nucleic acid sequence is at least 90% identical to nucleotides 61-1416 of SEQ ID NO:2, and wherein the second nucleic acid sequence is at least 90% identical to nucleotides 61-705 of SEQ ID NO:4.
21. The pharmaceutical composition of claim 4 , wherein the first nucleic acid sequence is 100% identical to nucleotides 61-426 of SEQ ID NO:2, and wherein the second nucleic acid sequence is 100% identical to nucleotides 61-384 of SEQ ID NO:4.
22. The pharmaceutical composition of claim 4 , wherein the first nucleic acid sequence is 100% identical to nucleotides 61-1416 of SEQ ID NO:2, and wherein the second nucleic acid sequence is 100% identical to nucleotides 61-705 of SEQ ID NO:4.
23. The pharmaceutical composition of claim 4 , wherein the first nucleic acid sequence is 100% identical to SEQ ID NO:2, and wherein the second nucleic acid sequence is 100% identical to SEQ ID NO:4.