Human cytomegalovirus gB polypeptide

Patent metadata

Field	Value
Publication number	US-12447207-B2
Application number	US-202318506578-A
Country	US
Kind code	B2
Filing date	Nov 10, 2023
Priority date	Jun 21, 2020
Publication date	Oct 21, 2025
Grant date	Oct 21, 2025

Abstract

Polypeptides and cytomegalovirus (CMV) antigens that include at least two introduced amino acid mutations relative to the amino acid sequence of the wild-type HCMV glycoprotein B (gB). For example, polypeptides stabilized in a conformation alternative to the gB postfusion conformation. Compositions including the polypeptides and uses thereof.

First claim

The invention claimed is:

1. A human cytomegalovirus (HCMV) immunogenic composition comprising: RNA formulated in a lipid nanoparticle,
wherein the RNA encodes an amino acid sequence for a glycoprotein B (gB) protein comprising at least one engineered disulfide mutation, wherein the gB protein comprises a cysteine mutation selected from the group consisting of: D217C, Y589C, M371C, W506C, N524C, and M684C, numbered according to SEQ ID NO:1.

2. The HCMV immunogenic composition of claim 1 , wherein the lipid nanoparticle comprises a cationic lipid, a polyethylene glycol (PEG)-modified lipid, a sterol, and a non-cationic lipid.

3. The HCMV immunogenic composition of claim 2 , wherein the cationic lipid is an ionizable cationic lipid, the non-cationic lipid is a neutral lipid, and the sterol is a cholesterol.

4. The HCMV immunogenic composition of claim 2 , wherein the cationic lipid is selected from the group consisting of: 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319).

5. The HCMV immunogenic composition of claim 2 , wherein the lipid nanoparticle has a molar ratio of about 20-60% cationic lipid, about 5-25% non-cationic lipid, about 25-55% sterol, and about 0.5-15% PEG-modified lipid.

6. The HCMV immunogenic composition of claim 1 , wherein the lipid nanoparticle has a polydispersity value of less than 0.4.

7. The HCMV immunogenic composition of claim 1 , wherein the lipid nanoparticle has a net neutral charge at a neutral pH.

8. The HCMV immunogenic composition of claim 1 , wherein the lipid nanoparticle has a mean diameter of 50-200 nm.

9. A human cytomegalovirus (HCMV) immunogenic composition comprising: RNA formulated in a lipid nanoparticle,
wherein the RNA encodes an amino acid sequence for a glycoprotein B (gB) protein comprising at least two amino acid mutations, and wherein said at least two amino acid mutations comprise:
a) at least one engineered disulfide mutation selected from the group consisting of: D217C and Y589C, and M371C and W506C, numbered according to SEQ ID NO:1; and
b) at least one additional mutation selected from the group consisting of:
i) substitution of YIH at positions 155-157 with GHR;
ii) substitution of W at position 240 with A;
iii) substitution of C at position 246 with S;
iv) substitution of P at position 655 with S;
v) substitution of F at position 678 with S;
vi) substitution of L at position 680 with T;
vii) substitution of R at position 685 with A;
viii) substitution of MIALDI (SEQ ID NO: 275) at positions 648-653 with GSGKDG (SEQ ID NO: 276);
ix) substitution of R at position 693 with V;
x) substitution of I at positions 767 and 768 with C;
xi) substitution of D at position 703 and P at position 704 with C; and
xii) substitution of Y at position 696 and V at position 697 with C,
wherein the positions are numbered according to SEQ ID NO: 1.

10. The HCMV immunogenic composition of claim 9 , wherein the lipid nanoparticle comprises a cationic lipid, a PEG-modified lipid, a sterol, and a non-cationic lipid.

11. The HCMV immunogenic composition of claim 10 , wherein the cationic lipid is an ionizable cationic lipid, the non-cationic lipid is a neutral lipid, and the sterol is a cholesterol.

12. The HCMV immunogenic composition of claim 10 , wherein the cationic lipid is selected from the group consisting of: 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319).

13. The HCMV immunogenic composition of claim 10 , wherein the lipid nanoparticle has a molar ratio of about 20-60% cationic lipid, about 5-25% non-cationic lipid, about 25-55% sterol, and about 0.5-15% PEG-modified lipid.

14. A human cytomegalovirus (HCMV) immunogenic composition comprising: RNA formulated in a lipid nanoparticle,
wherein the RNA encodes an amino acid sequence for a glycoprotein B (gB) protein comprising at least one engineered disulfide mutation selected from the group consisting of: D217C and Y589C, and M371C and W506C, numbered according to SEQ ID NO:1.

15. The HCMV immunogenic composition of claim 14 , wherein the gB protein comprises at least two engineered disulfide mutations.

16. The HCMV immunogenic composition of claim 14 , wherein the gB protein comprises an additional mutation selected from the group consisting of:
(1) substitution of YIH at positions 155-157 with GHR;
(2) substitution of W at position 240 with A; and
(3) substitution of C at position 246 with S, wherein the positions are numbered according to SEQ ID NO: 1.

17. The HCMV immunogenic composition of claim 14 , wherein the gB comprises the following mutations: D217C, Y589C, M371C, W506C, Y155G, I156H, H157R, W240A, and C246S, wherein the positions are numbered according to SEQ ID NO: 1.

18. The HCMV immunogenic composition of claim 14 , wherein the lipid nanoparticle comprises a cationic lipid, a PEG-modified lipid, a sterol, and a non-cationic lipid.

19. The HCMV immunogenic composition of claim 18 , wherein the cationic lipid is an ionizable cationic lipid, the non-cationic lipid is a neutral lipid, and the sterol is a cholesterol.

20. The HCMV immunogenic composition of claim 18 , wherein the lipid nanoparticle has a molar ratio of about 20-60% cationic lipid, about 5-25% non-cationic lipid, about 25-55% sterol, and about 0.5-15% PEG-modified lipid.

Assignees

• Pfizer

Inventors

• Liu Yuhang
• Che Ye
• Chi Xiaoyuan Sherry
• Dormitzer Philip Ralph
• Nicki Jennifer Anne
• Yao Xiaojie

Classifications

• C12N2710/16134

  Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein · CPC title

• C12N7/00

  Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00) · CPC title

• C07K14/045

  Cytomegalovirus · CPC title

• C12N2710/16122

  New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes · CPC title

• A61P31/22

  for herpes viruses · CPC title