Coronavirus vaccine formulations
Disclosed herein are coronavirus Spike (S) proteins and nanoparticles comprising the same, which are suitable for use in vaccines. The nanoparticles present antigens from pathogens surrounded to and associated with a detergent core resulting in enhanced stability and good immunogenicity. Dosages, formulations, and methods for preparing the vaccines and nanoparticles are also disclosed.
1 . A coronavirus (CoV) Spike(S) glycoprotein comprising:
(i) an S1 subunit comprising an inactivated furin cleavage site having the amino acid sequence of QQAQ (SEQ ID NO: 7); wherein the S1 subunit comprises an N-terminal domain (NTD); a receptor binding domain (RBD), and subdomains 1 and 2 (SD1/2);
wherein the N-terminal domain comprises up to about 20 modifications compared to a NTD of a CoV S glycoprotein of SEQ ID NO: 2;
wherein the RBD comprises up to about 30 modifications compared to a RBD of a CoV S glycoprotein of SEQ ID NO: 2;
wherein the SD1/2 comprises up to about 15 modifications compared to a SD1/2 of a CoV S glycoprotein of SEQ ID NO: 2; and
(ii) an S2 subunit, wherein amino acids 973 and 974 are proline,
wherein the S2 subunit comprises up to 15 additional modifications compared to the S2 subunit of a CoV S glycoprotein of SEQ ID NO: 2;
wherein the amino acids of the CoV S glycoprotein are numbered with respect to the CoV S glycoprotein of SEQ ID NO: 2;
wherein each modification is a mutation, deletion, or addition of one amino acid.
2 . The CoV S glycoprotein of claim 1 , wherein the NTD of the CoV S glycoprotein comprises between about 5 and about 10 modifications compared to the NTD of SEQ ID NO: 2.
3 . The CoV S glycoprotein of claim 1 , wherein the NTD of the CoV S glycoprotein comprises between about 8 and about 12 modifications compared to the NTD of SEQ ID NO: 2.
4 . The CoV S glycoprotein of claim 1 , wherein the RBD of the CoV S glycoprotein comprises between about 18 and about 23 modifications compared to the RBD of SEQ ID NO: 2.
5 . The CoV S glycoprotein of claim 1 , wherein the RBD of the CoV S glycoprotein comprises between about 12 and about 17 modifications compared to the RBD of SEQ ID NO: 2.
6 . The CoV S glycoprotein of claim 1 , wherein the RBD of the CoV S glycoprotein comprises between about 5 and about 10 modifications compared to the RBD of SEQ ID NO: 2.
7 . The CoV S glycoprotein of claim 1 , wherein the RBD of the CoV S glycoprotein comprises between about 15 and about 20 modifications compared to the RBD of SEQ ID NO: 2.
8 . The CoV S glycoprotein of claim 1 , wherein the SD1/2 of the CoV S glycoprotein comprises between about 3 and about 10 modifications compared to the SD1/2 of SEQ ID NO: 2.
9 . The CoV S glycoprotein of claim 1 , wherein the S2 subunit of the CoV S glycoprotein comprises between about 3 and about 10 modifications compared to the S2 subunit of SEQ ID NO: 2.
10 . The CoV S glycoprotein of claim 1 , wherein the CoV S glycoprotein comprises one or more modifications at a position selected from the group consisting of 56, 57, 131, 404, 488, 439, 471, 601, 642, and 668, wherein the CoV S glycoprotein is numbered according to SEQ ID NO: 2.
11 . A nanoparticle comprising the CoV S glycoprotein of claim 1 and a non-ionic detergent.
12 . The nanoparticle of claim 11 , wherein the non-ionic detergent is selected from the group consisting of polysorbate-20 (PS20), polysorbate-40 (PS40), polysorbate-60 (PS60), polysorbate-65 (PS65), and polysorbate-80 (PS80).
13 . An immunogenic composition comprising the nanoparticle of claim 11 and a pharmaceutically acceptable buffer.
14 . The immunogenic composition of claim 13 , comprising an adjuvant.
15 . The immunogenic composition of claim 14 , wherein the adjuvant is a saponin adjuvant.
16 . The immunogenic composition of claim 13 , wherein the saponin adjuvant comprises at least two iscom particles, wherein:
the first iscom particle comprises fraction A of Quillaja Saponaria Molina and not fraction C of Quillaja Saponaria Molina; and
the second iscom particle comprises fraction C of Quillaja Saponaria Molina and not fraction A of Quillaja Saponaria Molina.
17 . The immunogenic composition of claim 16 , wherein fraction A of Quillaja Saponaria Molina accounts for 50-96% by weight and fraction C of Quillaja Saponaria Molina accounts for the remainder, respectively, of the sum of the weights of fraction A of Quillaja Saponaria Molina and fraction C of Quillaja Saponaria Molina in the adjuvant.
18 . The immunogenic composition of claim 16 , wherein fraction A of Quillaja Saponaria Molina and fraction C of Quillaja Saponaria Molina account for about 85% by weight and about 15% by weight, respectively, of the sum of the weights of fraction A of Quillaja Saponaria Molina and fraction C of Quillaja Saponaria Molina in the adjuvant.
19 . The immunogenic composition of claim 13 , comprising from about 3 μg to about 25 μg of CoV S glycoprotein.
20 . The immunogenic composition of claim 13 , comprising about 5 μg of CoV S glycoprotein.
21 . The immunogenic composition of claim 14 , comprising about 50 μg of adjuvant.
22 . A method of stimulating an immune response against SARS-COV-2 in a human comprising administering the immunogenic composition of claim 13 .
23 . The method of claim 22 , wherein the immunogenic composition comprises an adjuvant.
24 . The method of claim 23 , wherein the adjuvant is a saponin adjuvant.
25 . The method of claim 24 , wherein the saponin adjuvant comprises at least two iscom particles, wherein:
the first iscom particle comprises fraction A of Quillaja Saponaria Molina and not fraction C of Quillaja Saponaria Molina; and
the second iscom particle comprises fraction C of Quillaja Saponaria Molina and not fraction A of Quillaja Saponaria Molina.
26 . The method of claim 25 , wherein fraction A of Quillaja Saponaria Molina accounts for 50-96% by weight and fraction C of Quillaja Saponaria Molina accounts for the remainder, respectively, of the sum of the weights of fraction A of Quillaja Saponaria Molina and fraction C of Quillaja Saponaria Molina in the adjuvant.
27 . The method of claim 25 , wherein fraction A of Quillaja Saponaria Molina and fraction C of Quillaja Saponaria Molina account for about 85% by weight and about 15% by weight, respectively, of the sum of the weights of fraction A of Quillaja Saponaria Molina and fraction C of Quillaja Saponaria Molina in the adjuvant.
28 . The method of claim 22 , comprising from about 3 μg to about 25 μg of CoV S glycoprotein.
29 . The method of claim 22 , comprising about 5 μg of CoV S glycoprotein.
30 . The method of claim 23 , comprising about 50 μg of adjuvant.