Patent Application
20240261392
While vaccines targeting the SARS-COV-2 spike protein have been successful in slowing the spread of COVID-19 infections, recent data suggests that immunity conferred by these vaccines may decrease over time. Furthermore, instances of breakthrough infections have been reported, suggesting that the SARS-COV-2 virus may be capable of evading the host immune response. There is a need for vaccines that confer enhanced immunity to SARS-COV-2.
In various aspects, the present disclosure provides a composition comprising: a nucleic acid molecule encoding a coronavirus ORF8 peptide, and a lipid particle encapsulating the nucleic acid molecule.
In some aspects, the lipid particle comprises lipids selected from the group consisting of 2[(polyethylene glycol (PEG))-2000]-N,N-ditetradecylacetamide; 1,2-distearoyl-sn-glycero-3-phosphocholine; cholesterol; ((4-hydroxybutyl)azanediyl)bis(hexane-6,1-diyl)bis(2-hexyldecanoate); 1,2-dimyristoyl-rac-glycerol, methoxypolyethylene glycol; heptadecane-9-yl 8-((2-hydroxyethyl) (6-oxo-6-(undecyloxy) hexyl) amino) octanoate; and combinations thereof.
In various aspects, the present disclosure provides a composition comprising: a nucleic acid molecule encoding a coronavirus ORF8 peptide, and a viral vector encapsulating the nucleic acid molecule.
In some aspects, the viral vector is an attenuated viral vector. In some aspects, the viral vector is an adenovirus vector. In some aspects, the adenovirus vector is a replication-deficient adenovirus. In some aspects, the replication-deficient adenovirus is a ChAd0x1 adenovirus, an Ad5 adenovirus, or an Ad26 adenovirus. In some aspects, the viral vector is an attenuated coronavirus.
In some aspects, the nucleic acid molecule is an RNA molecule. In some aspects, the nucleic acid molecule is a DNA molecule. In some aspects, the nucleic acid molecule further encodes an additional coronavirus protein comprising a coronavirus spike protein or an immunogenic fragment thereof, a coronavirus envelope protein or an immunogenic fragment thereof, a coronavirus membrane protein or an immunogenic fragment thereof, a coronavirus nucleocapsid protein or an immunogenic fragment thereof, or combinations thereof. In some aspects, the nucleic acid molecule codes for expression of the coronavirus ORF8 peptide in a subject upon administration of the composition to the subject.
In various aspects, the present disclosure provides a composition comprising: a coronavirus ORF8 peptide, and an adjuvant capable of enhancing an immune response upon administration to a subject.
In some aspects, the adjuvant comprises an aluminum salt, a monophosphorylated lipid A, or a cytosine phosphoguanine. In some aspects, the composition further comprises a carrier protein fused to the coronavirus ORF8 peptide. In some aspects, the carrier protein is adsorbed onto the adjuvant.
In some aspects, the composition further comprises an additional immunogenic peptide. In some aspects, the nucleic acid molecule further encodes an additional immunogenic peptide. In some aspects, the additional immunogenic peptide is an additional coronavirus peptide or an influenza peptide. In some aspects, the additional coronavirus peptide comprises a coronavirus spike protein or an immunogenic fragment thereof, a coronavirus envelope protein or an immunogenic fragment thereof, a coronavirus membrane protein or an immunogenic fragment thereof, or a coronavirus nucleocapsid protein or an immunogenic fragment thereof.
In some aspects, the coronavirus ORF8 peptide comprises an ORF8 protein or an immunogenic fragment thereof. In some aspects, the coronavirus ORF8 peptide comprises a sequence having at least 80% sequence identity to any one of SEQ ID NO: 1 or SEQ ID NO: 9-SEQ ID NO: 11. In some aspects, the coronavirus ORF8 peptide comprises a sequence of any one of SEQ ID NO: 1 or SEQ ID NO: 9-SEQ ID NO: 11. In some aspects, the coronavirus ORF8 peptide comprises from 6 to 120 consecutive amino acid residues of a sequence having at least 80% sequence identity to any one of SEQ ID NO: 1 or SEQ ID NO: 9-SEQ ID NO: 11. In some aspects, the coronavirus ORF8 peptide comprises from 6 to 120 consecutive residues of a sequence of any one of SEQ ID NO: 1 or SEQ ID NO: 9-SEQ ID NO: 11. In some aspects, the coronavirus ORF8 peptide comprises a sequence of any one of SEQ ID NO: 5-SEQ ID NO: 8 or SEQ ID NO: 12-SEQ ID NO: 20.
In some aspects, the composition further comprises a salt, a sugar, a stabilizer, or combinations thereof. In some aspects, the salt comprises sodium phosphate, potassium phosphate, potassium chloride, sodium chloride, sodium acetate, acetic acid, trisodium citrate, citric acid, or combinations thereof. In some aspects, the sugar comprises sucrose, 2-hydroxypropyl-β-cyclodextrin, or combinations thereof. In some aspects, the stabilizer comprises tromethamine, tromethamine hydrochloride, polysorbate-80, ethanol, or combinations thereof. In some aspects, the composition is formulated for intramuscular injection.
In various aspects, the present disclosure provides a method of inducing an immune response in a subject, the method comprising: administering to the subject a composition comprising a nucleic acid molecule encoding a coronavirus ORF8 peptide; delivering the nucleic acid molecule to a cell of the subject; expressing the coronavirus ORF8 peptide in the cell; and inducing an immune response to the coronavirus ORF8 peptide.
In various aspects, the present disclosure provides a method of inducing an immune response in a subject, the method comprising: administering to the subject a composition comprising a coronavirus ORF8 peptide; and inducing an immune response to the coronavirus ORF8 peptide.
In various aspects, the present disclosure provides a method of inducing an immune response in a subject, the method comprising: administering to the subject a composition described herein; and inducing an immune response to the coronavirus ORF8 peptide.
In some aspects, the method further comprises administering an additional coronavirus vaccine to the subject within 90 days of administering the composition. In some aspects, the additional coronavirus vaccine is a SARS-COV vaccine or a SARS-COV-2 vaccine. In some aspects, the method further comprises administering an influenza vaccine to the subject within 90 days of administering the composition. In some aspects, the additional coronavirus vaccine, the influenza vaccine, or both is administered within one day of the composition. In some aspects, the additional coronavirus vaccine, the influenza vaccine, or both is administered within 15 minutes of the composition. In some aspects, the additional coronavirus vaccine, the influenza vaccine, or both is formulated with the composition.
In some aspects, the subject is a previously immunized subject. In some aspects, the subject is a previously infected subject. In some aspects, the subject has partial immunity to the coronavirus. In some aspects, the partial immunity is conferred by a vaccination state or an infection state.
In some aspects, the method further comprises enhancing an immune response to the coronavirus relative to administration of a vaccine lacking the ORF peptide or a vaccine lacking the nucleic acid molecule encoding the ORF8 peptide. In some aspects, the method further comprises reducing the risk of infection by the coronavirus in the subject upon exposure of the subject to the coronavirus. In some aspects, the method further comprises reducing immune invasion of the coronavirus in the subject upon exposure of the subject to the coronavirus. In some aspects, the method further comprises viral entry of the coronavirus in the subject upon exposure of the subject to the coronavirus. In some aspects, the method further comprises reducing viral replication of the coronavirus in the subject upon exposure of the subject to the coronavirus. In some aspects, the method further comprises preventing inhibition of MHC class I by the coronavirus upon exposure of the subject to the coronavirus.
In some aspects, the coronavirus is a sarbecovirus. In some aspects, the coronavirus is SARS-COV or SARS-COV-2. In some aspects, the coronavirus encodes ORF8. In some aspects, the coronavirus inhibits viral antigen presentation in an infected cell.
All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.
The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings of which:
The present disclosure provides compositions and methods for increasing an immune response of a subject to a coronavirus (e.g., a SARS-COV-2 coronavirus). SARS-COV-2 is a highly virulent strain of coronavirus that causes COVID-19 which killed more than 6 million people and infected more than 500 million between December of 2019 and July of 2022. SARS-CoV-2 is the seventh coronavirus known to infect humans; SARS-COV, MERS-COV and SARS-CoV-2 can cause severe disease, whereas HKU1, NL63, OC43 and 229E are associated with mild symptoms. The SARS-COV-2 human coronavirus is a 29,903-nucleotide, positive-strand RNA virus that is associated with a variety of highly prevalent and severe diseases, including SARS and Middle East respiratory syndrome (MERS). While the vaccines targeting the SARS-CoV-2 spike protein substantially reduce the risk of infection, data suggests that the immunity conferred by these vaccines may diminish over time and, in some instances, breakthrough infections may occur. Described herein are methods for increasing an immune response of a subject to a coronavirus by inoculating the subject with a composition comprising a coronavirus ORF8 protein, an immunogenic fragment of an ORF8 protein, or a nucleic acid encoding an ORF8 protein or immunogenic fragment of an ORF8 protein.
The SARS-COV-2 open reading frame 8 (ORF8) protein is a 121-amino acid protein comprising an N-terminal signal sequence followed by a predicted Ig-like fold. The SARS-COV-2 ORF8 protein forms a homodimer connected by a single disulfide bond. The structure of SARS-COV-2 ORF8 is shown in
SARS-COV-2 ORF8 has an amino acid sequence of MKFLVFLGIITTVAAFHQECSLQSCTQHQPYVVDDPCPIHFYSKWYIRVGARKSAPLIEL CVDEAGSKSPIQYIDIGNYTVSCLPFTINCQEPKLGSLVVRCSFYEDFLEYHDVRVVLDFI (SEQ ID NO: 1). The SARS-COV-2 ORF8 sequence is highly divergent from other coronavirus ORF8 sequences, sharing less than 20% sequence identity to SARS-COV ORF8. This divergence in the ORF8 sequence may contribute to the increased virulence of SARS-COV-2 relative to other coronaviruses since genetic divergence often contributes to increased virulence in new viral strains. The most conserved region in the ORF8 protein of SARS-COV-2 is PFTINCQE (SEQ ID NO: 5) which is present in the catalytic core of the protein. An amino acid sequence alignment of SARS-COV-2 ORF8 to RaTG13 ORF8, a coronavirus that infects horseshoe bats, is shown in
ORF8 is a coronavirus accessory protein may interfere with host inflammatory responses and may contribute to immune evasion of SARS-COV-2, as illustrated in
The ORF8 protein of SARS-COV-2 induces endoplasmic reticulum stress and mediated immune evasion by antagonizing production of interferon β. Both SARS-COV-2 ORF8 genotypes, ORF8L and ORF8S, induced ER stress pathways, antagonize interferon β production, and decrease the nuclear translocation of IRF3 induced by poly. The SARS-COV-2 ORF8 mainly acts as an immune-modulator by down-regulating MHC class I molecules, thereby shielding the infected cells against cytotoxic T cells, killing the target cells. The ORF8 also regulates unfolded protein response (UPR) induced due to the ER stress by triggering the ATF-6 activation, thus enhancing the survivability of infected cells. ORF8 may inhibit induction intracellular aggregation, lysosomal stress, and interleukin-mediated inflammatory responses by activating NLRP3 inflammasomes, thereby inhibiting the host immune response to.
SARS-COV-2 coronaviruses containing an ORF8 deletion are associated with COVID-19 infections with milder symptoms compared to COVID-19 infections caused by SARS-COV-2 coronaviruses with a wild type ORF8. The 382-nucleotide deletion variant (Δ382) causes the deletion of the entire ORF8 protein. Patients with the Δ382 variant exhibit less severe symptoms, including milder hypoxic conditions and low cytokine activity compared to patients infected with the wild type virus. The Δ382 variant shows reduced viral replication ability in human cells, suggesting that SARS-COV-2 ORF8 functions in transmission and viral replication efficiency.
A composition for boosting an immune response of a subject to a coronavirus (e.g., a SARS-COV-2 coronavirus) may comprise a coronavirus ORF8 protein (e.g., SEQ ID NO: 1), an immunogenic fragment of a coronavirus ORF8 protein (e.g., a fragment of SEQ ID NO: 1 comprising from 6 to 120 amino acid residues), a nucleic acid (e.g., a DNA molecule or an RNA molecule) encoding a coronavirus ORF8 protein, or a nucleic acid encoding a fragment of a coronavirus ORF8 protein. The coronavirus may be SARS-COV, SARS-COV-2, MERS-COV, HKU1, OC43, or 229E. The coronavirus may be a beta-coronavirus.
In some embodiments, the ORF8 protein comprises a sequence of SEQ ID NO: 1. In some embodiments, the ORF8 protein may be an ORF8 variant comprising a sequence having at least 80%, 85%, 90%, 95%, or 99% sequence identity to SEQ ID NO: 1. For example, the ORF8 variant protein may comprise one or more substitutions selected from V62L, L84S, or S24L, relative to SEQ ID NO: 1. In some embodiments, the ORF8 protein comprises a fragment of a sequence having at least 70%, 80%, 85%, 90%, 95%, or 99% sequence identity to SEQ ID NO: 1 or SEQ ID NO: 9-SEQ ID NO: 11. For example, the ORF8 protein fragment may comprise a fragment of an ORF8 protein comprising one or more substitutions selected from V62L, L84S, or S24L, relative to SEQ ID NO: 1. In some embodiments, the ORF8 variant may be an ORF8 protein found in a coronavirus variant (e.g., an ORF8 from a SARS-COV-2 a, B, y, or 8 variant). In some embodiments, the ORF8 variant may be an ORF8 variation found within a coronavirus strain. In some embodiments, an ORF8 variant may be an engineered ORF8. An ORF8 variant may be engineered to have structural homology to wild type ORF8 or to an ORF8 protein found in a coronavirus variant. Structural homology may be determined using bioinformatic software (e.g., ROSETTA, IntFOLD, RaptorX, Biskit, ESyPred3D, Phyre, MODELLER, or the like). Antibodies generated against the engineered ORF8 variant may also recognize and bind wild type ORF8 or an ORF8 from a coronavirus variant. In some embodiments, an ORF8 variant may comprise a V62L substitution (e.g.,
An ORF8 protein fragment may be an immunogenic fragment of an ORF8 protein capable of triggering an immune response against the ORF8 fragment or full-length ORF8 in a subject. Antibodies generated against the ORF8 fragment may also recognize and bind full-length ORF8. An immunogenic fragment of ORF8 may be a surface-exposed fragment of ORF8. An immunogenic fragment of ORF8 may be identified using bioinformatics software (e.g., ExPASy, Protean, or BLASTN) to identify fragments predicted to have high immunogenicity. In some embodiments, an immunogenic fragment may be identified by selecting for fragments of ORF8 that bind to one or more polyclonal antibodies generated against full-length ORF8. In some embodiments, an ORF8 fragment (e.g., an immunogenic fragment) may be an ORF8 truncation found in a coronavirus strain or variant. In some embodiments, an ORF8 fragment may be a fragment shown in TABLE 1.
In some embodiments, an ORF8 fragment may comprise a sequence of VDEAGSKS (SEQ ID NO: 7) or DEAGSKS (SEQ ID NO: 8). An ORF8 fragment may be a fragment of a variant ORF8 sequence having one or more mutations relative to SEQ ID NO: 1. For example, an ORF8 fragment may be a fragment of an ORF8 sequence comprising one or more substitutions selected from V62L, L84S, or S24L relative to SEQ ID NO: 1. In some embodiments, an ORF8 fragment may comprise from 6 to 120 amino acid residues of any one of SEQ ID NO: 1 or SEQ ID NO: 9-SEQ ID NO: 11 or a sequence having at least 70%, 80%, 85%, 90%, 95%, or 99% sequence identity to any one of SEQ ID NO: 1 or SEQ ID NO: 9-SEQ ID NO: 11. The ORF8 fragment may be a surface-exposed fragment of an ORF8 peptide (e.g., a peptide of any one of SEQ ID NO: 1 or SEQ ID NO: 9-SEQ ID NO: 11). In some embodiments, an ORF8 fragment may comprise a sequence of any one of SEQ ID NO: 5-SEQ ID NO: 8 or SEQ ID NO: 12-SEQ ID NO: 20.
In some embodiments, an immunogenic fragment of an ORF8 peptide may have a length of from about 6 to about 20, from about 20 to about 30, from about 30 to about 40, from about 40 to about 50, from about 50 to about 60, from about 60 to about 70, from about 70 to about 80, from about 80 to about 90, from about 90 to about 100, from about 100 to about 110, from about 110 to about 120, from about 6 to about 30, from about 6 to about 50, from about 6 to about 75, from about 6 to about 100, from about 6 to about 120, from about 10 to about 30, from about 10 to about 50, from about 10 to about 75, from about 10 to about 100, from about 10 to about 120, from about 20 to about 50, from about 20 to about 75, from about 20 to about 100, from about 20 to about 120, from about 50 to about 75, from about 50 to about 100, or from about 50 to about 120 amino residues.
In some embodiments, a composition may comprise an ORF8 protein, a fragment of an ORF8 protein, or a nucleic acid encoding an ORF8 protein or protein fragment encapsulated by a delivery vehicle. In some embodiments, the delivery vehicle may comprise a lipid capsid, a viral vector, or an attenuated virus. For example, the delivery vehicle may be a lipid nanoparticle, an adenovirus vector, or an attenuated coronavirus. The delivery vehicle may facilitate delivery of the ORF8 protein, protein fragment, or coding sequence to a host cell.
A composition of the present disclosure may be formulated as an ORF8 vaccine for delivery to a subject. An ORF8 vaccine may comprise an ORF8 protein (e.g., any one of SEQ ID NO: 1 or SEQ ID NO: 9-SEQ ID NO: 11) or an immunogenic fragment of an ORF8 protein (e.g., any one of SEQ ID NO: 5-SEQ ID NO: 8 or SEQ ID NO: 12-SEQ ID NO: 20), or an ORF8 vaccine may comprise a nucleic acid encoding for an ORF8 protein (e.g., any one of SEQ ID NO: 1 or SEQ ID NO: 9-SEQ ID NO: 11) or an immunogenic fragment of an ORF8 protein (e.g., any one of SEQ ID NO: 5-SEQ ID NO: 8 or SEQ ID NO: 12-SEQ ID NO: 20). For example, a composition may be formulated as an mRNA vaccine comprising an mRNA encoding an ORF8 protein or protein fragment encapsulated in a lipid particle. In another example, a composition may be formulated an adenovirus vaccine comprising a DNA molecule encoding an ORF8 protein or protein fragment encapsulated in an adenovirus vector. In another example, a composition may be formulated as a subunit vaccine, also referred to as an epitope vaccine, comprising an ORF8 protein or protein fragment.
In some embodiments, an ORF8 vaccine may be formulated in a lipid particle. The ORF8 vaccine may comprise a nucleic acid molecule encoding an ORF8 protein or an immunogenic fragment thereof and a lipid particle encapsulating the nucleic acid molecule, wherein the ORF8 protein is an coronavirus ORF8 protein. The lipid particle may encapsulate a nucleic acid molecule encoding for an ORF8 protein (e.g., any one of SEQ ID NO: 1 or SEQ ID NO: 9-SEQ ID NO: 11) or an immunogenic fragment of an ORF8 protein (e.g., any one of SEQ ID NO: 5-SEQ ID NO: 8 or SEQ ID NO: 12-SEQ ID NO: 20). The nucleic acid molecule may be DNA or RNA (e.g., mRNA). In some embodiments, the lipid particle may comprise lipids selected from the group consisting of 2 [(polyethylene glycol (PEG))-2000]-N,N-ditetradecylacetamide; 1,2-distearoyl-sn-glycero-3-phosphocholine; cholesterol; ((4-hydroxybutyl)azanediyl)bis(hexane-6,1-diyl)bis(2-hexyldecanoate); 1,2-dimyristoyl-rac-glycerol, methoxypolyethylene glycol; heptadecane-9-yl 8-((2-hydroxyethyl) (6-oxo-6-(undecyloxy) hexyl) amino) octanoate; and combinations thereof. For example, the lipid particle may comprise 2[(polyethylene glycol (PEG))-2000]-N,N-ditetradecylacetamide, 1,2-distearoyl-sn-glycero-3-phosphocholine, cholesterol, and ((4-hydroxybutyl)azanediyl)bis(hexane-6,1-diyl)bis(2-hexyldecanoate). In another example, the lipid particle may comprise 1,2-dimyristoyl-rac-glycerol, methoxypolyethylene glycol, 1,2-distearoyl-sn-glycero-3-phosphocholine, cholesterol, and heptadecane-9-yl 8-((2-hydroxyethyl) (6-oxo-6-(undecyloxy) hexyl) amino) octanoate. The ORF8 vaccine comprising a lipid particle encapsulating a nucleic acid molecule may further comprise a salt (e.g., sodium phosphate, potassium phosphate, potassium chloride, sodium chloride, sodium acetate, acetic acid, trisodium citrate, citric acid, or combinations thereof), a sugar (e.g., sucrose, 2-hydroxypropyl-β-cyclodextrin, or combinations thereof), a stabilizer (e.g., tromethamine, tromethamine hydrochloride, polysorbate-80, ethanol, or combinations thereof), or a combination thereof. For example, the ORF8 vaccine may comprise sodium phosphate, potassium phosphate, potassium chloride, sodium chloride, and sucrose. In another example, the ORF8 vaccine may comprise sodium acetate, sucrose, tromethamine, tromethamine hydrochloride, and acetic acid.
In some embodiments, an ORF8 vaccine may be formulated as a viral vector. The ORF8 vaccine may comprise a nucleic acid molecule encoding an ORF8 protein or an immunogenic fragment thereof, and a viral vector encapsulating the nucleic acid molecule, wherein the ORF8 protein is a coronavirus ORF8 protein. The viral vector may encapsulate a nucleic acid molecule encoding for an ORF8 protein (e.g., any one of SEQ ID NO: 1 or SEQ ID NO: 9-SEQ ID NO: 11) or an immunogenic fragment of an ORF8 protein (e.g., any one of SEQ ID NO: 5-SEQ ID NO: 8 or SEQ ID NO: 12-SEQ ID NO: 20). The nucleic acid molecule may be DNA or RNA. The viral vector may be an attenuated virus (e.g., an attenuated coronavirus) or an adenovirus (e.g., a replication-deficient adenovirus). In some embodiments, the adenovirus may be a ChAd0x1 adenovirus, an Ad5 adenovirus, or an Ad26 adenovirus. The ORF8 vaccine comprising a viral vector encapsulating a nucleic acid molecule may further comprise a salt (e.g., sodium phosphate, potassium phosphate, potassium chloride, sodium chloride, sodium acetate, acetic acid, trisodium citrate, citric acid, or combinations thereof), a sugar (e.g., sucrose, 2-hydroxypropyl-β-cyclodextrin, or combinations thereof), a stabilizer (e.g., tromethamine, tromethamine hydrochloride, polysorbate-80, ethanol, or combinations thereof), or a combination thereof. For example, the ORF8 vaccine may comprise polysorbate 80,2-hydroxypropyl-β-cyclodextrin, trisodium citrate, sodium chloride, citric acid, and ethanol.
In some embodiments, an ORF8 vaccine may be formulated as a subunit or epitope vaccine. The ORF8 vaccine may comprise an ORF8 protein or an immunogenic fragment thereof and an adjuvant capable of enhancing an immune response upon administration to a subject, wherein the ORF8 protein is a coronavirus ORF8 protein. The ORF8 vaccine may comprise an ORF8 protein (e.g., any one of SEQ ID NO: 1 or SEQ ID NO: 9-SEQ ID NO: 11) or an immunogenic fragment of an ORF8 protein (e.g., any one of SEQ ID NO: 5-SEQ ID NO: 8 or SEQ ID NO: 12-SEQ ID NO: 20). The adjuvant may comprise an aluminum salt, a monophosphorylated lipid A, or a cytosine phosphoguanine. For example, the adjuvant may be aluminum hydroxide. In some embodiments, the ORF8 vaccine may further comprise a carrier protein fused to the ORF8 protein or the immunogenic fragment. The ORF8 vaccine comprising a viral vector encapsulating a nucleic acid molecule may further comprise a salt (e.g., sodium phosphate, potassium phosphate, potassium chloride, sodium chloride, sodium acetate, acetic acid, trisodium citrate, citric acid, or combinations thereof), a sugar (e.g., sucrose, 2-hydroxypropyl-β-cyclodextrin, or combinations thereof), a stabilizer (e.g., tromethamine, tromethamine hydrochloride, polysorbate-80, ethanol, or combinations thereof), or a combination thereof.
An ORF8 vaccine (e.g., a vaccine comprising an ORF8 peptide or immunogenic fragment or a nucleic acid encoding an ORF8 peptide or immunogenic fragment) may enhance an immune response to a viral infection (e.g., a coronavirus infection) when administered alone or in combination with an additional vaccine (e.g., a coronavirus vaccine or an influenza vaccine). The ORF8 vaccine may induce production of anti-ORF8 antibodies in a subject upon administration of the vaccine. The anti-ORF8 antibodies may facilitate an immune response to a virus (e.g., an ORF8-encoding coronavirus). In some embodiments, the ORF8 vaccine may increase T cell killing of cells infected with an ORF8-encoding virus (e.g., an ORF8-encoding coronavirus). In some embodiments, the ORF8 vaccine may increase T cell killing of cells with inhibited viral antigen presentation (e.g., cells with downregulated MHC-1).
An ORF8 vaccine may be formulated in combination with an additional immunogenic agent, such as a coronavirus spike protein, a fragment of a coronavirus spike protein, a membrane protein or protein fragment, an envelope protein or protein fragment, or a nucleocapsid protein or protein fragment, or a nucleic acid encoding a coronavirus spike protein or spike protein fragment, a membrane protein or protein fragment, an envelope protein or protein fragment, or a nucleocapsid protein or protein fragment. In some embodiments, the composition may be formulated as a supplemental or booster vaccine to boost immunity conferred by a coronavirus vaccine.
A method for boosting an immune response of a subject to a coronavirus (e.g., a SARS-CoV-2 coronavirus) may comprise administering a ORF8 vaccine composition of the present disclosure (e.g., a composition comprising a coronavirus ORF8 protein, an immunogenic fragment of a coronavirus ORF8 protein, or a nucleic acid encoding a coronavirus ORF8 protein or immunogenic fragment of a coronavirus ORF8 protein) to a subject. The coronavirus may be SARS-COV, SARS-COV-2, MERS-COV, HKU1, OC43, or 229E. The coronavirus may be a beta-coronavirus. The coronavirus may be a sarbecovirus. The coronavirus may be an ORF8-encoding coronavirus. The composition may induce an immune response in the subject, producing antibodies against the ORF8 protein. The anti-ORF8 antibodies may facilitate an immune response to a virus (e.g., an ORF8-encoding coronavirus). In some embodiments, the ORF8 vaccine may increase T cell killing of cells infected with an ORF8-encoding virus (e.g., an ORF8-encoding coronavirus). In some embodiments, the ORF8 vaccine may increase T cell killing of cells with inhibited viral antigen presentation (e.g., cells with downregulated MHC-1).
In some embodiments, an ORF8 vaccine composition may be administered as a booster following a coronavirus vaccine (e.g., an mRNA vaccine, an adenovirus vaccine, or a subunit vaccine designed to trigger an immune response to a coronavirus spike protein). In some embodiments, an ORF8 vaccine composition may be formulated as a single vaccine with a coronavirus vaccine. For example, single vaccine formulation may comprise a nucleic acid encoding both an ORF8 protein or protein fragment and a spike protein or spike protein fragment, a membrane protein or protein fragment, an envelope protein or protein fragment, or a nucleocapsid protein or protein fragment. In some embodiments, an ORF8 vaccine composition may be administered concurrently with a coronavirus vaccine. An ORF8 vaccine composition of the present disclosure may be formulated in combination with an additional vaccine or immunogenic agent. For example, the ORF8 vaccine may be formulated in combination with a coronavirus vaccine (e.g., targeting a spike protein) or an influenza vaccine. An ORF8 vaccine composition of the present disclosure may be administered in combination with an additional vaccine. For example, the ORF8 vaccine may administered in combination with a coronavirus vaccine (e.g., targeting a spike protein) or an influenza vaccine. In some embodiments an ORF8 vaccine may be administered as a booster to a previously-vaccinated individual (e.g., an individual who had previously received a coronavirus vaccine) or to a previously-infected individual (e.g., an individual previously infected with a coronavirus).
An ORF8 vaccine may be administered in combination with an additional vaccine (e.g., a coronavirus vaccine or an influenza vaccine). The ORF8 vaccine may be administered within about 6 months, about 5 months, about 4 months, about 3 months, about 90 days, about 60 days, about 30 days, about 15 days, about 2 weeks, about 10 days, about 1 week, about 5 days, about 4 days, about 3 days, about 3 days, about 1 day, about 12 hours, about 6 hours, about 4 hours, about 2 hours, about 1 hour, about 30 minutes, about 15 minutes, about 5 minutes, or about 1 minute of an additional vaccine (e.g., a coronavirus vaccine or an influenza vaccine). In some embodiments, the ORF8 vaccine may be formulated in combination with the additional vaccine.
Administration of an ORF8 vaccine composition may reduce the risk of a coronavirus infection in the subject. In some embodiments, administration of an ORF8 vaccine composition may reduce immune invasion of the coronavirus in the subject. Administration of an ORF8 vaccine composition may reduce viral entry of a coronavirus into a cell of a subject or reduce viral replication of the coronavirus in the subject. In some embodiments, administration of an ORF8 vaccine composition may prevent inhibition of MHC-I. Administration of the composition may enhance immunity of the subject to a coronavirus by inducing production of antibodies against ORF8 in the subject. The antibodies against ORF8 may enhance immunity of the subject by binding to ORF8 produced by an infecting coronavirus and degrading or inhibiting the ORF8, thereby blocking the immune-evading function of ORF8.
As used herein, the terms “about” and “approximately,” in reference to a number, is used herein to include numbers that fall within a range of 10%, 5%, or 1% in either direction (greater than or less than) the number unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value).
One of ordinary skill will appreciate that the less than (“<”) and greater than (“>”) symbols or terminology used herein can be replaced with less than or equal to (“<”) and greater than or equal to (“>”) symbols, respectively, without departing from the scope of this description.
The invention is further illustrated by the following non-limiting examples.
This example describes administration of an ORF8 mRNA vaccine as a booster to enhance immunity to a coronavirus. An mRNA encoding an ORF8 protein is encapsulated in a lipid particle. The lipid-encapsulated ORF8 mRNA is formulated as a ORF8 mRNA vaccine for administration to a subject. The subject is a previously-vaccinated subject or a previously-infected subject and already has some level of immunity to the coronavirus. Upon administration of the ORF8 mRNA vaccine to the subject, the ORF8 mRNA enters a cell of the subject and is transcribed into ORF8 protein. The subject generates antibodies against the ORF8 protein, thereby enhancing the immunity of the subject against the coronavirus.
This example describes administration of a combination ORF8 mRNA vaccine to enhance immunity to a coronavirus. An mRNA encoding an ORF8 protein and one or more additional coronavirus proteins or protein fragments, such as a coronavirus spike protein, is encapsulated in a lipid particle. The lipid-encapsulated mRNA is formulated as an mRNA vaccine for administration to a subject. Upon administration of the mRNA vaccine to the subject, the mRNA enters a cell of the subject and ORF8 protein is transcribed by the host cell. The subject generates antibodies against the ORF8 protein, thereby enhancing the immunity of the subject against the coronavirus compared to administration of an mRNA vaccine lacking ORF8 mRNA.
This example describes administration of an ORF8 adenovirus vaccine as a booster to enhance immunity to a coronavirus. A DNA encoding an ORF8 protein is encapsulated in an adenovirus vector. The adenovirus-encapsulated ORF8 DNA is formulated as an ORF8 adenovirus vaccine for administration to a subject. The subject is a previously-vaccinated subject or a previously-infected subject and already has some level of immunity to the coronavirus. Upon administration of the ORF8 adenovirus vaccine to the subject, the ORF8 DNA enters a cell of the subject and is transcribed into ORF8 protein. The subject generates antibodies against the ORF8 protein, thereby enhancing the immunity of the subject against the coronavirus.
This example describes administration of a combination ORF8 adenovirus vaccine to enhance immunity to a coronavirus. A DNA encoding an ORF8 protein and one or more additional coronavirus proteins or protein fragments, such as a coronavirus spike protein, is encapsulated in an adenovirus vector. The adenovirus-encapsulated DNA is formulated as an adenovirus vaccine for administration to a subject. Upon administration of the adenovirus vaccine to the subject, the DNA enters a cell of the subject and ORF8 protein is transcribed by the host cell. The subject generates antibodies against the ORF8 protein, thereby enhancing the immunity of the subject against the coronavirus compared to administration of an adenovirus vaccine lacking ORF8 DNA.
This example describes administration of an ORF8 protein vaccine as a booster to enhance immunity to a coronavirus. An ORF8 protein is formulated as an ORF8 protein vaccine for administration to a subject. The subject is a previously-vaccinated subject or a previously-infected subject and already has some level of immunity to the coronavirus. Upon administration of the ORF8 adenovirus vaccine to the subject, the subject generates antibodies against the ORF8 protein, thereby enhancing the immunity of the subject against the coronavirus.
This example describes administration of a combination ORF8 protein vaccine to enhance immunity to a coronavirus. An ORF8 protein and one or more additional coronavirus proteins or protein fragments, such as a coronavirus spike protein, are formulated as a protein vaccine for administration to a subject. Upon administration of the adenovirus vaccine to the subject, the subject generates antibodies against the ORF8 protein, thereby enhancing the immunity of the subject against the coronavirus compared to administration of a protein vaccine lacking ORF8 DNA.
This example describes formulation of a lipid particle ORF8 vaccine. An RNA molecule encoding an ORF8 peptide (e.g., an ORF8 protein of any one of SEQ ID NO: 1 or SEQ ID NO: 9-SEQ ID NO: 11 or an immunogenic fragment of any one of SEQ ID NO: 5-SEQ ID NO: 8 or SEQ ID NO: 12-SEQ ID NO: 20 is encapsulated in a lipid particle. Optionally, the RNA molecule further encodes a coronavirus spike protein or an immunogenic fragment thereof, a coronavirus envelope protein or an immunogenic fragment thereof, a coronavirus membrane protein or an immunogenic fragment thereof, a coronavirus nucleocapsid protein or an immunogenic fragment thereof. The lipid particle comprises 2[(polyethylene glycol (PEG))-2000]-N,N-ditetradecylacetamide, 1,2-distearoyl-sn-glycero-3-phosphocholine, cholesterol, and ((4-hydroxybutyl)azanediyl)bis(hexane-6,1-diyl)bis(2-hexyldecanoate). The vaccine formulation further comprises sodium phosphate, potassium phosphate, potassium chloride, sodium chloride, and sucrose. The ORF8 vaccine is formulated for intramuscular administration.
This example describes an additional formulation of a lipid particle ORF8 vaccine. An RNA molecule encoding an ORF8 peptide (e.g., an ORF8 protein of any one of SEQ ID NO: 1 or SEQ ID NO: 9-SEQ ID NO: 11 or an immunogenic fragment of any one of SEQ ID NO: 5-SEQ ID NO: 8 or SEQ ID NO: 12-SEQ ID NO: 20 is encapsulated in a lipid particle. Optionally, the RNA molecule further encodes a coronavirus spike protein or an immunogenic fragment thereof, a coronavirus envelope protein or an immunogenic fragment thereof, a coronavirus membrane protein or an immunogenic fragment thereof, a coronavirus nucleocapsid protein or an immunogenic fragment thereof. The lipid particle comprises 1,2-dimyristoyl-rac-glycerol, methoxypolyethylene glycol, 1,2-distearoyl-sn-glycero-3-phosphocholine, cholesterol, and heptadecane-9-yl 8-((2-hydroxyethyl) (6-oxo-6-(undecyloxy) hexyl) amino) octanoate. The vaccine formulation further comprises sodium acetate, sucrose, tromethamine, tromethamine hydrochloride, and acetic acid. The ORF8 vaccine is formulated for intramuscular administration.
This example describes formulation of an adenovirus ORF8 vaccine. A DNA molecule encoding an ORF8 peptide (e.g., an ORF8 protein of any one of SEQ ID NO: 1 or SEQ ID NO: 9-SEQ ID NO: 11 or an immunogenic fragment of any one of SEQ ID NO: 5-SEQ ID NO: 8 or SEQ ID NO: 12-SEQ ID NO: 20 is encapsulated in an adenovirus vector. Optionally, the DNA molecule further encodes a coronavirus spike protein or an immunogenic fragment thereof, a coronavirus envelope protein or an immunogenic fragment thereof, a coronavirus membrane protein or an immunogenic fragment thereof, a coronavirus nucleocapsid protein or an immunogenic fragment thereof. The adenovirus is an Ad26 adenovirus. The vaccine formulation further comprises polysorbate 80,2-hydroxypropyl-β-cyclodextrin, trisodium citrate, sodium chloride, citric acid, and ethanol. The ORF8 vaccine is formulated for intramuscular administration.
This example describes formulation of an epitope ORF8 vaccine. The vaccine comprises an ORF8 peptide (e.g., an ORF8 protein of any one of SEQ ID NO: 1 or SEQ ID NO: 9-SEQ ID NO: 11 or an immunogenic fragment of any one of SEQ ID NO: 5-SEQ ID NO: 8 or SEQ ID NO: 12-SEQ ID NO: 20 fused to a carrier protein. Optionally, the vaccine further comprises a coronavirus spike protein or an immunogenic fragment thereof, a coronavirus envelope protein or an immunogenic fragment thereof, a coronavirus membrane protein or an immunogenic fragment thereof, a coronavirus nucleocapsid protein or an immunogenic fragment thereof fused to a carrier protein. The ORF8 peptide fused to the carrier protein is adsorbed to an aluminum hydroxide adjuvant. The vaccine formulation further comprises sodium chloride. The ORF8 vaccine is formulated for intramuscular administration.
While preferred embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.
The present application claims the benefit of U.S. Provisional Application No. 63/220,389, entitled “COMPOSITIONS AND METHODS TO INCREASE CORONAVIRUS IMMUNE RESPONSE,” filed on Jul. 9, 2021, which application is herein incorporated by reference in its entirety for all purposes.
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/US2022/036471 | 7/8/2022 | WO |
| Number | Date | Country | |
|---|---|---|---|
| 63220389 | Jul 2021 | US |
