Coronavirus vaccine
The present invention is directed to a nucleic acid suitable for use in treatment or prophylaxis of an infection with a coronavirus, preferably with a Coronavirus SARS-CoV-2, or a disorder related to such an infection, preferably COVID-19. The present invention is also directed to compositions, polypeptides, and vaccines. The compositions and vaccines preferably comprise at least one of said nucleic acid sequences, preferably nucleic acid sequences in association a lipid nanoparticle (LNP). The invention is also directed to first and second medical uses of the nucleic acid, the composition, the polypeptide, the combination, the vaccine, and the kit, and to methods of treating or preventing a coronavirus infection, preferably a Coronavirus infection.
1. A composition comprising a mRNA comprising:
(a) at least one coding sequence encoding a SARS-CoV-2 spike protein (S) at least 95% identical to SEQ ID NO: 10 that is a pre-fusion stabilized spike protein (S_stab) comprising a pre-fusion stabilizing K986P and V987P mutation;
(b) at least one heterologous untranslated region (UTR); and
(c) at least one pharmaceutically acceptable carrier,
wherein the mRNA is complexed or associated with lipid nanoparticles (LNP) and wherein the LNP comprises:
(i) at least one cationic lipid according to formula III-3:
(ii) at least one neutral lipid, comprising 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC);
(iii) at least one steroid, comprising cholesterol; and
(iv) at least one PEG-lipid according to formula IVa:
wherein n has a mean value ranging from 30 to 60,
wherein (i) to (iv) are in a molar ratio of about 20-60% cationic lipid, 5-25% neutral lipid, 25-55% sterol, and 0.5-15% PEG-lipid.
2. The composition of claim 1 , wherein the mRNA comprises at least one poly(A) sequence comprising 30 to 200 adenosine nucleotides.
3. The composition of claim 1 , wherein the mRNA comprises a 5′-cap structure.
4. The composition of claim 1 , wherein the at least one coding sequence of the mRNA is a codon modified coding sequence, wherein the at least one codon modified coding sequence is selected from C maximized coding sequence, CAI maximized coding sequence, human codon usage adapted coding sequence, G/C content modified coding sequence, and G/C optimized coding sequence, or any combination thereof.
5. The composition of claim 1 , wherein the at least one coding sequence of the mRNA has a G/C content that is increased by at least 10% compared to the G/C content of the coding sequence of the corresponding wild-type or reference nucleic acid sequence.
6. The composition of claim 1 , wherein the mRNA comprises a sequence at least 90% identical to SEQ ID NO: 163.
7. The composition of claim 1 , wherein the at least one heterologous untranslated region is selected from at least one heterologous 5′-UTR and/or at least one heterologous 3′-UTR.
8. The composition of claim 7 , wherein the at least one heterologous 3′-UTR comprises or consists of a nucleic acid sequence derived from a 3′-UTR of a gene selected from PSMB3, ALB7, alpha-globin, CASP1, COX6B1, GNAS, NDUFA1 and RPS9.
9. The composition of claim 7 , wherein the at least one heterologous 5′-UTR comprises or consists of a nucleic acid sequence derived from a 5′-UTR of a gene selected from HSD17B4, RPL32, ASAH1, ATP5A1, MP68, NDUFA4, NOSIP, RPL31, SLC7A3, TUBB4B and UBQLN2.
10. The composition of claim 1 , wherein the nucleic acid comprises at least one histone stem-loop.
11. The composition of claim 1 , wherein the mRNA comprises a nucleotide analog.
12. The composition of claim 1 , wherein the mRNA does not comprise a 1-methylpseudouridine substitution.
13. The composition of claim 1 , wherein the mRNA is a purified mRNA that has been purified by RP-HPLC and/or TFF.
14. The composition of claim 1 , wherein the mRNA is a purified mRNA that has been purified by TFF.
15. The composition of claim 1 , wherein the LNP comprises at least one PEG-lipid according to formula (IVa) wherein n has a mean value ranging from about 40±2, 42±2, 44±2, 46±2, 48±2, 50±2, 52±2, 54±2, 56±2 or 58±2.
16. The composition of claim 1 , wherein the LNPs have a mean diameter of from about 30 nm, 35 nm, 40 nm, 45 nm, 50 nm, 55 nm, 60 nm, 65 nm, 70 nm, 75 nm, 80 nm, 85 nm, 90 nm, 95 nm, 100 nm, 105 nm, 110 nm, 115 nm, 120 nm, 125 nm, 130 nm, 135 nm, 140 nm, 145 nm, 150 nm, 160 nm, 170 nm, 180 nm, 190 nm to 200 nm.
17. The composition of claim 3 , wherein 5′-cap structure comprises a m7G, cap0, cap1, cap2, a modified cap0 or a modified cap1 structure.
18. The composition of claim 11 , wherein the mRNA comprises a 1-methylpseudouridine substitution.
19. The composition of claim 1 , further comprising a lyoprotectant.
20. The composition of claim 19 , wherein the lyoprotectant comprises sucrose.
21. The composition of claim 1 , wherein the composition has a lipid to RNA molar ratio (N/P ratio) of from about 2 to about 12.
22. The composition of claim 1 , wherein the LNP comprises a molar ratio of approximately 47.4:10:40.9:1.7 of cationic lipid:DSPC:cholesterol:PEG.
23. The composition of claim 1 , wherein the at least one coding sequence encoding a SARS-CoV-2 spike protein (S) is identical to SEQ ID NO: 10.
24. The composition of claim 6 , wherein the mRNA comprises a sequence at least 92% identical to SEQ ID NO: 163.
25. The composition of claim 24 , wherein the mRNA comprises a sequence at least 95% identical to SEQ ID NO: 163.
26. The composition of claim 1 , wherein the mRNA comprises a sequence at least 80% identical to SEQ ID NO: 137.
27. The composition of claim 26 , wherein the mRNA comprises a sequence at least 85% identical to SEQ ID NO: 137.