Vaccination in newborns and infants

Abstract

The present invention relates to vaccines comprising at least one mRNA encoding at least one antigen for use in the treatment of a disease in newborns and/or infants, preferably exhibiting an age of not more than 2 years, preferably of not more than 1 year, more preferably of not more than 9 months or even 6 months, wherein the treatment comprises vaccination of the newborn or infant and eliciting an immune response in said newborn or infant. The present invention is furthermore directed to kits and kits of parts comprising such a vaccine and/or its components and to methods applying such a vaccine or kit.

Claims

1. A method for stimulating a protective antigen specific immune response in an infant subject comprising, administering to the subject a first effective amount of a composition comprising a purified mRNA encoding a viral antigen, said mRNA comprising a 5′ cap and a polyA tail, wherein said mRNA is complexed with a cationic or polycationic compound, wherein the subject is an infant subject being no more than 2 years of age and wherein the composition is administered by injection and wherein the viral antigen is an influenza hemagglutinin (HA) virus antigen, or an antigenic fragment thereof.

2. The method of claim 1, wherein the subject is about 3 months to 2 years of age.

3. The method of claim 2, wherein the subject is no more than 1 year of age.

4. The method of claim 3, wherein the subject is no more than 6 months of age.

5. The method of claim 1, wherein the composition is administered by intradermal or intramuscular injection.

6. The method of claim 5, wherein the composition is administered by intramuscular injection.

7. The method of claim 1, wherein the cationic or polycationic compound comprises a cationic lipid.

8. The method of claim 1, wherein the mRNA comprises a backbone modification, a sugar modification, or a base modification.

9. The method of claim 8, wherein the mRNA comprises a base modification.

10. The method of claim 1, further comprising administering an adjuvant to the subject.

11. The method of claim 10, wherein the adjuvant is an immunostimulatory RNA (isRNA).

12. The method of claim 1, wherein the antigen-specific immune response comprises production of antigen-specific antibodies.

13. The method of claim 12, wherein the antigen-specific immune response comprises an antigen-specific Th1 immune response.

14. The method of claim 1, wherein the composition further comprises a mRNA encoding at least a second viral antigen or an antigenic fragment thereof.

15. The method of claim 1, further comprising administering at least a second dose of the composition to the subject.

16. The method of claim 15, wherein the second dose of the composition is administered at least 10 days after the first administration.

17. The method of claim 1, wherein the composition further comprises polyethyleneglycol.

18. A method for stimulating a protective antigen specific immune response in an infant subject comprising, administering to the subject an effective amount of a composition comprising a purified mRNA encoding, a viral antigen, said mRNA comprising: a 5′ cap, a polyA tail, and at least one base modification, said composition comprising a cationic lipid, wherein the subject is an infant subject being no more than 2 years of age and wherein the composition is administered by intramuscular injection wherein the viral antigen is an influenza hemagglutinin (HA) virus antigen, or an antigenic fragment thereof.

19. The method of claim 1, wherein the G/C content of the mRNA encoding the viral antigen is increased compared to an original RNA sequence encoding the antigen.

20. The method of claim 19, wherein the G/C content of the mRNA encoding the viral antigen is increased by at least 15% compared to an original RNA sequence encoding the antigen.

21. The method of claim 1, wherein the cationic or polycationic compound is a polypeptide.

22. The method of claim 21, wherein the polypeptide is protamine.

23. The method of claim 1, wherein the viral antigen is full length HA.

24. The method of claim 18, wherein the viral antigen is a full length HA virus antigen.

25. The method of claim 18, wherein the G/C content of the mRNA encoding the viral antigen is increased compared to an original RNA sequence encoding the antigen.

26. The method of claim 25, wherein the G/C content of the mRNA encoding the viral antigen is increased by at least 15% compared to an original RNA sequence encoding the antigen.

27. The method of claim 18, wherein the subject is about 3 months to 2 years of age.

28. The method of claim 27, wherein the subject is no more than 1 year of age.

29. The method of claim 18, wherein the subject is no more than 6 months of age.

30. The method of claim 27, wherein the composition further comprises polyethyleneglycol.

31. The method of claim 18, wherein the composition further comprises a mRNA encoding at least a second viral antigen or an antigenic fragment thereof.

Description

FIGURES

(1) The following Figures are intended to illustrate the invention further. They are not intended to limit the subject matter of the invention thereto.

(2) FIG. 1 A: shows the development of the weight of the mice in the experiment. As a result newborn mice vaccinated with mRNA coding for PR8 H1 Hemagglutinin exhibited a significantly better survival (all mice survived) against influenza challenge infection with control mRNA only (all mice died about 5 days subsequent to vaccination with control mRNA encoding chicken ovalbumin, when vaccinated with control mRNA at the first day 8 weeks and died about 6 days subsequent to vaccination with control mRNA, when vaccinated with 8 weeks). Most surprisingly, the survival rate was comparable to that of adult mice.

(3) FIGS. 1 B, C show the coding sequence of the mRNAs used for vaccination of newborn and 8 weeks old mice (see FIG. 1A) coding for PR8 H1 HA (Hemagglutinin of influenza virus A/Puerto Rico/8/1934) (SEQ ID NO: 384) (FIG. 1B) or for Gallus gallus ovalbumine as a control (control mRNA) (SEQ ID NO: 385) (FIG. 1C)

EXAMPLES

(4) The following examples are intended to illustrate the invention further. They are not intended to limit the subject matter of the invention thereto.

Example 1—Preparation of mRNA Constructs

(5) For the present examples DNA sequences, encoding PR8 H1 HA (Haemagglutinin of A/Puerto Rico/8/1934) (SEQ ID NO: 384), and Gallus gallus ovalbumine, respectively, as a control (control mRNA) (SEQ ID NO: 385), were prepared and used for subsequent in vitro transcription reactions.

(6) According to a first preparation, the DNA sequence termed PR8 H1 HA (Haemagglutinin of A/Puerto Rico/8/1934) (SEQ ID NO: 384) (see FIG. 1B) was prepared by modifying the wildtype Haemagglutinin encoding DNA sequence by introducing a GC-optimized sequence for a better codon usage and stabilization. In SEQ ID NO: 384 (see FIG. 1B) the sequence of the corresponding mRNA is shown. The sequence was furthermore introduced into a pCV19 vector and modified to comprise stabilizing sequences derived from alpha-globin-3′-UTR (muag (mutated alpha-globin-3′-UTR)), a stretch of 70×adenosine at the 3′-terminal end (poly-A-tail) and a stretch of 30×cytosine at the 3′-terminal end (poly-C-tail). The sequence of the final DNA construct was termed “PR8 H1 HA”.

(7) According to a second preparation, the DNA sequence termed Gallus gallus ovalbumine, respectively, as a control (control mRNA) (SEQ ID NO: 385) (see FIG. 1C) was prepared by modifying the wildtype Gallus gallus ovalbumine encoding DNA sequence by introducing a GC-optimized sequence for a better codon usage and stabilization. In SEQ ID NO: 385 (see FIG. 1C) the sequence of the corresponding mRNA is shown. The sequence was furthermore introduced into a pCV19 vector and modified to comprise stabilizing sequences derived from alpha-globin-3′-UTR (muag (mutated alpha-globin-3′-UTR)), a stretch of 70×adenosine at the 3′-terminal end (poly-A-tail) and a stretch of 30×cytosine at the 3′-terminal end (poly-C-tail). The sequence of the final DNA construct was termed “Gallus gallus ovalbumine”.

(8) In a further step, the respective DNA plasmids prepared above were transcribed into mRNA in vitro using T7-Polymerase. Subsequently the obtained mRNA was purified using PUREMESSENGER® (CureVac, Tubingen, Germany).

(9) All obtained mRNAs used herein were furthermore complexed with protamine prior to use. The RNA complexation consisted of a mixture of 50% free mRNA and 50% mRNA complexed with protamine at a weight ratio of 2:1. First, mRNA was complexed with protamine by slow addition of protamine-Ringer's lactate solution to mRNA. As soon as the complexes were stably generated, free mRNA was added, stirred shortly and the final concentration of the vaccine was adjusted with Ringer's lactate solution.

Example 2—Vaccination of Newborn and 8 Weeks Old Mice

(10) In this experiment newborn or 8 weeks old mice were vaccinated twice intradermally with 80 μg mRNA coding for PR8 H1 HA (Hemagglutinin of A/Puerto Rico/8/1934; FIG. 1B) or with mRNA coding for Gallus gallus ovalbumine as a control (control mRNA; FIG. 1C). The first injection was carried out at the first day of life (≤24 h) and with 8 weeks, respectively. 5 weeks after the last vaccination the mice were challenged with a 10fold median lethal dose of PR8 virus (10 LD50). The weight of the mice was controlled over 2 weeks and the mice were killed when they have lost more than 25% of their original weight. The results are shown in FIG. 1A. FIG. 1A shows the development of the weight of the mice in the experiment. As a result mice vaccinated with mRNA coding for PR8 H1 Hemagglutinin exhibited a significantly better survival (all mice survived) against influenza challenge infection with control mRNA only (all mice in the control experiment died about 5 days subsequent to vaccination with control mRNA encoding chicken ovalbumin, when vaccinated with control mRNA at the first day and died about 6 days subsequent to vaccination with control mRNA, when vaccinated with 8 weeks). All vaccinated newborn mice survived antigen challenge with PR8 H1 Hemagglutinin in contrast to the control.