Multisubunit papillomavirus vaccines and therapeutics

Abstract

The present disclosure relates generally to multisubunit nucleic acids comprising a plurality of self-assembling immunogenic sequences or a plurality of self-assembling immunogenic sequences each further comprising one or more target sequence, or a combination thereof, wherein each self-assembling immunogenic sequence of the plurality is connected to an adjacent self-assembling immunogenic sequence of the plurality by a cleavage sequence, and wherein the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the self-assembling immunogenic sequences of the plurality. Also disclosed are multisubunit nucleic acids comprising a plurality of polynucleotide sequences, wherein some or all polynucleotide sequences of the plurality comprises a plurality of self-assembling immunogenic sequence, or a plurality of self-assembling immunogenic sequence each further comprising one or more target sequence, or a combination thereof, wherein each polynucleotide sequence of the plurality is connected to an adjacent polynucleotide sequence of the plurality by one or more cleavage sequence and wherein the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the polynucleotide sequence of the plurality. The multisubunit nucleic acid encodes multisubunit peptide.

Description

[0001] Multisubunit Papillomavirus Vaccines and Therapeutics

[0002] RELATED APPLICATIONS

[0003] This application claims the benefit of priority to U.S. Provisional Patent Application serial number 63 / 665,025, filed on June 27, 2024, the contents of which are hereby incorporated herein by reference in its entirety.

[0004] REFERENCE TO A SEQUENCE LISTING XML

[0005] This application contains a Sequence Listing which has been submitted electronically in XML format. The Sequence Listing XML is incorporated herein by reference. Said XML file, created on June 23, 2025, is named PVM-00525.xml and is 10,866,448 bytes in size.

[0006] BACKGROUND

[0007] Human papillomaviruses (HPVs) are the most prominent papillomaviruses which are known to cause papillomas or warts, and are responsible for causing cervical cancer, vaginal cancer, vulvar cancer, penile cancer, anal cancer, and oropharyngeal cancer. Of the approximate 46,000 cases of cancer per year in the United States, about 37,000 of them are caused by human papillomaviruses. According to 2020 estimates, global incidence of cancer cases attributed to HPV are around 730,000, of them a significant number (660,000) are found in women. Prophylactic HPV vaccines have been available since 2006, but vaccine-associated HPV type-replacement is a concern. Further, prophylactic HPV vaccines do not provide effective treatment for an already infected person. Therefore, effective vaccines and therapeutics against HPV are required.

[0008] SUMMARY

[0009] Accordingly, the present disclosure relates to a multisubunit nucleic acid comprising either a plurality of self- assembling immunogenic sequences or a plurality of self-assembling immunogenic sequences each further comprising one or more target sequence, or a combination thereof, wherein each self- assembling immunogenic sequence is connected to an adjacent self-assembling immunogenic sequence of the plurality by one or more cleavage sequence, and wherein the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the self-assembling immunogenic sequences of the plurality.

[0010] In another aspect, provided herein is a multisubunit nucleic acid comprising a plurality of self- assembling immunogenic sequences, wherein each self-assembling immunogenic sequence of the plurality is connected to an adjacent self-assembling immunogenic sequence of the plurality by one or more cleavage sequence. In some embodiments, the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the self-assembling immunogenic sequences of the plurality.

[0011] In another aspect, provided herein is a multisubunit nucleic acid comprising a plurality of self- assembling immunogenic sequences, wherein each self-assembling immunogenic sequence further comprises one or more target sequence, wherein each selfassembling immunogenic sequence of the plurality is connected to an adjacent selfassembling immunogenic sequence of the plurality by one or more cleavage sequence. In some embodiments, the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the self-assembling immunogenic sequences of the plurality.

[0012] In another aspect, provided herein is a vaccine comprising a multisubunit nucleic acid, wherein the multisubunit nucleic acid comprises either a plurality of self- assembling immunogenic sequences or a plurality of self-assembling immunogenic sequences each further comprising one or more target sequence, or a combination thereof, wherein each self-assembling immunogenic sequence is connected to an adjacent self-assembling immunogenic sequence of the plurality by one or more cleavage sequence, and wherein the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the self-assembling immunogenic sequences of the plurality.

[0013] In another aspect, provided herein is a vaccine comprising a multisubunit nucleic acid, wherein the multisubunit nucleic acid comprises a plurality of self-assembling immunogenic sequences, wherein each self-assembling immunogenic sequence of the plurality is connected to an adjacent self-assembling immunogenic sequence of the plurality by one or more cleavage sequence. In some embodiments, the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the self-assembling immunogenic sequences of the plurality.

[0014] In another aspect, provided herein is a vaccine comprising a multisubunit nucleic acid, wherein the multisubunit nucleic acid comprises a plurality of self-assembling immunogenic sequences, wherein each self-assembling immunogenic sequence further comprises one or more target sequence, wherein each self-assembling immunogenic sequence of the plurality is connected to an adjacent self-assembling immunogenic sequence of the plurality by one or more cleavage sequence. In some embodiments, the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the self-assembling immunogenic sequences of the plurality.

[0015] In another aspect, provided herein is a multisubunit nucleic acid comprising a plurality of polynucleotide sequences, wherein some or all polynucleotide sequences of the plurality comprises either a self-assembling immunogenic sequence, or a self-assembling immunogenic sequence further comprising one or more target sequence, or a combination thereof, wherein each polynucleotide sequence of the plurality is connected to an adjacent polynucleotide sequence of the plurality by one or more cleavage sequence. In some embodiments, the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the polynucleotide sequences of the plurality.

[0016] In another embodiment, provided herein is a multisubunit nucleic acid comprising a plurality of polynucleotide sequences, wherein each polynucleotide sequence of the plurality comprises a self-assembling immunogenic sequence, wherein each polynucleotide sequence of the plurality is connected to an adjacent polynucleotide sequence of the plurality by one or more cleavage sequence, and wherein the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the polynucleotide sequences of the plurality.

[0017] In another embodiment, provided herein is a multisubunit nucleic acid comprising a plurality of polynucleotide sequences, wherein each polynucleotide sequence of the plurality comprises a self-assembling immunogenic sequence further comprising one or more target sequence, wherein each polynucleotide sequence of the plurality is connected to an adjacent polynucleotide sequence of the plurality by one or more cleavage sequence, and wherein the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the polynucleotide sequences of the plurality.

[0018] In another aspect, provided herein is a vaccine comprising a multisubunit nucleic acid, wherein the multisubunit nucleic acid comprises a plurality of polynucleotide sequences, wherein some or all polynucleotide sequences of the plurality comprises either a self-assembling immunogenic sequence, or a self-assembling immunogenic sequence further comprising one or more target sequence, or a combination thereof, wherein each polynucleotide sequence of the plurality is connected to an adjacent polynucleotide sequence of the plurality by one or more cleavage sequence. In some embodiments, the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the polynucleotide sequences of the plurality.

[0019] In another embodiment, provided herein is a vaccine comprising a multisubunit nucleic acid, wherein the multisubunit nucleic acid comprises a plurality of polynucleotide sequences, wherein each polynucleotide sequence of the plurality comprises a selfassembling immunogenic sequence, wherein each polynucleotide sequence of the plurality is connected to an adjacent polynucleotide sequence of the plurality by one or more cleavage sequence, and wherein the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the polynucleotide sequences of the plurality.

[0020] In another embodiment, provided herein is a vaccine comprising a multisubunit nucleic acid, wherein the multisubunit nucleic acid comprises a plurality of polynucleotide sequences, wherein each polynucleotide sequence of the plurality comprises a selfassembling immunogenic sequence further comprising one or more target sequence, wherein each polynucleotide sequence of the plurality is connected to an adjacent polynucleotide sequence of the plurality by one or more cleavage sequence, and wherein the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the polynucleotide sequences of the plurality.

[0021] In another aspect, provided herein is a multisubunit nucleic acid encoding a multisubunit peptide, wherein the multisubunit peptide comprises either a plurality of selfassembling immunogenic peptides or a plurality of self-assembling immunogenic peptides each further comprising one or more target peptide, or a combination thereof, wherein each self-assembling immunogenic peptide is connected to an adjacent self-assembling immunogenic peptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide upstream of one or more of the self-assembling immunogenic peptides of the plurality.

[0022] In another aspect, provided herein is a multisubunit nucleic acid encoding a multisubunit peptide, wherein the multisubunit peptide comprises a plurality of selfassembling immunogenic peptides, wherein each self-assembling immunogenic peptide of the plurality is connected to an adjacent self-assembling immunogenic peptide of the plurality by one or more cleavage peptide. In some embodiments, the multisubunit peptide further comprises one or more signal peptide upstream (amino-terminus or N-terminus) of one or more of the self-assembling immunogenic peptides of the plurality.

[0023] In another aspect, provided herein is a multisubunit nucleic acid encoding a multisubunit peptide, wherein the multisubunit peptide comprises a plurality of selfassembling immunogenic peptides, wherein each self-assembling immunogenic peptide further comprises one or more target peptide, wherein each self-assembling immunogenic peptide of the plurality is connected to an adjacent self-assembling immunogenic peptide of the plurality by one or more cleavage peptide. In some embodiments, the multisubunit peptide further comprises one or more signal peptide upstream (amino-terminus or N- terminus) of one or more of the self-assembling immunogenic peptides of the plurality.

[0024] In another aspect, provided herein is a vaccine comprising a multisubunit nucleic acid encoding a multisubunit peptide, wherein the multisubunit peptide comprises either a plurality of self-assembling immunogenic peptides or a plurality of self-assembling immunogenic peptides each further comprising one or more target peptide, or a combination thereof, wherein each self-assembling immunogenic peptide is connected to an adjacent self-assembling immunogenic peptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide upstream of one or more of the self-assembling immunogenic peptide of the plurality.

[0025] In another aspect, provided herein is a vaccine comprising a multisubunit nucleic acid encoding a multisubunit peptide, wherein the multisubunit peptide comprises a plurality of self-assembling immunogenic peptides, wherein each self-assembling immunogenic peptide of the plurality is connected to an adjacent self-assembling immunogenic peptide of the plurality by one or more cleavage peptide. In some embodiments, the multisubunit peptide further comprises one or more signal peptide upstream (amino-terminus or N-terminus) of one or more of the self-assembling immunogenic peptides of the plurality.

[0026] In another aspect, provided herein is a vaccine comprising a multisubunit nucleic acid encoding a multisubunit peptide, wherein the multisubunit peptide comprises a plurality of self-assembling immunogenic peptides, wherein each self-assembling immunogenic peptide further comprises one or more target peptide, wherein each selfassembling immunogenic peptide of the plurality is connected to an adjacent selfassembling immunogenic peptide of the plurality by one or more cleavage peptide. In some embodiments, the multisubunit peptide further comprises one or more signal peptide upstream (amino-terminus or N-terminus) of one or more of the self-assembling immunogenic peptides of the plurality.

[0027] In another aspect, provided herein is a multisubunit nucleic acid encoding a multisubunit peptide comprising a plurality of polypeptides, wherein some or all polypeptides of the plurality comprises either a self-assembling immunogenic peptide, or a self-assembling immunogenic peptide each further comprising one or more target peptide, or a combination thereof, wherein each polypeptide of the plurality is connected to an adjacent polypeptide of the plurality by one or more cleavage peptide. In some embodiments, the polypeptide further comprises one or more signal peptide on the aminoterminus of one or more of the polypeptides of the plurality.

[0028] In another aspect, provided herein is a vaccine comprising a multisubunit nucleic acid encoding a multisubunit peptide, wherein the multisubunit peptide comprises a plurality of polypeptides, wherein some or all polypeptides of the plurality comprises either a self-assembling immunogenic peptide, or a self-assembling immunogenic peptide each further comprising one or more target peptide, or a combination thereof, wherein each polypeptide of the plurality is connected to an adjacent polypeptide of the plurality by one or more cleavage peptide. In some embodiments, the polypeptide further comprises one or more signal peptide on the amino-terminus of one or more of the polypeptides of the plurality.

[0029] In some embodiments, total number of the self-assembling immunogenic sequences or the polynucleotide sequences in a multisubunit nucleic acid are not more than 100. In some embodiments, total number of the self-assembling immunogenic sequences or the polynucleotide sequences in a multisubunit nucleic acid are between 2-5, 2-10, 10- 20, 20-30, 30-40, 40-50, 50-60, 60-70, 70-80, 80-90, or 90-99.

[0030] In some embodiments, the multisubunit nucleic acid is a DNA or an RNA. In some embodiments, the RNA is an mRNA. In some embodiments, the mRNA is obtained or synthesized through a single IVT process or step.

[0031] In some embodiments, the linker sequence encodes a linker peptide. In some embodiments, the linker peptide is an amino acid linker, a foldon, a scaffold or a combination thereof. In some embodiments, the linker peptide is an amino acid linker. In some embodiments, the linker peptide is a foldon. In some embodiments, the linker peptide is a scaffold. In some embodiments, the linker peptide comprises an amino acid linker and a foldon. In some embodiments, the linker peptide comprises an amino acid linker and a scaffold. In some embodiments, the linker peptide comprises a foldon and a scaffold. In some embodiments, the linker peptide comprises an amino acid linker, a foldon, and a scaffold. In some embodiments, the amino acid linker comprises 2 to 49 amino acids. In some embodiments, the amino acid linker is a glycine serine linker, a glycine proline linker, a glycine threonine linker, an alanine serine linker, any combination of two amino acids, or a combination thereof. In some embodiments, the linker peptide has an amino acid sequence of any one of SEQ ID NOs: 1-40.

[0032] In some embodiments, the cleavage sequence encodes a cleavage peptide. In some embodiments, the cleavage sequence encodes one or more cleavage peptide. In some embodiments, the cleavage peptide comprises one or more cleavage peptides, for example, cleavage peptide- 1, cleavage peptide-2, and so on. In some embodiments, the one or more cleavage peptides are optionally connected to each other by a linker peptide. In some embodiments, the cleavage peptide is a golgi specific cleavage peptide, self cleaving peptide, or a combination thereof. In some embodiments, the cleavage peptide has an amino acid sequence of any one of SEQ ID NOs: 41-55.

[0033] In some embodiments, the signal sequence encodes a signal peptide. In some embodiments, one or more signal peptide is present on the amino-terminus of one or more self-assembling immunogenic peptide or the polypeptide. In some embodiments, the multisubunit nucleic acid further encodes a second signal peptide on the amino-terminus of all or some self-assembling immunogenic peptides or polypeptides. In some embodiments, the signal peptide has an amino acid sequence of any one of SEQ ID NOs: 56-75.

[0034] In some embodiments, the self-assembling immunogenic sequence encodes a selfassembling immunogenic peptide. In some embodiments, the self-assembling immunogenic peptide is obtained or derived from LI protein of a papillomavirus, including their codon optimized nucleic acid sequences, fragments, mutants, variants, comparable equivalents, or functional analogs thereof. In some embodiments, the selfassembling immunogenic peptide has an amino acid sequence of any one of SEQ ID NOs: 76-1129 or 9409-9444, including its codon optimized nucleic acid sequence, or fragments, mutants, variants, comparable equivalents, or functional analogs thereof.

[0035] In some embodiments, the LI protein comprises one or more mutations, for example, C175A or C428A or both compared to SEQ ID NO: 712.

[0036] In some embodiments, the target sequence encodes a target peptide. In some embodiments, the target peptide is encoded by a codon optimized nucleic acid sequence, or fragments, mutants, or variants thereof. In some embodiments, the target peptide is obtained or derived from L2 protein, El protein, E2 protein, E4 protein, E5 protein, E6 protein, E7 protein, B cell epitope, T cell epitope, or a combination thereof of a papillomavirus, including their codon optimized nucleic acid sequences, or fragments, mutants, variants, comparable equivalents, or functional analogs thereof.

[0037] In some embodiments, the target peptide is inserted into the self-assembling immunogenic peptide. In some embodiments, the self-assembling immunogenic peptide is obtained or derived from LI protein. In some embodiments, the LI protein is obtained or derived from a papillomavirus.

[0038] In some embodiments, the target peptide is inserted into or within the BC loop, DE loop, EF loop, FG loop, HI loop, or a combination thereof of the LI protein of a papillomavirus.

[0039] In some embodiments, the papillomavirus is selected from the group comprising bovine papillomavirus (BPV), canine oral papillomavirus (COPV), cotton tail rabbit papillomavirus (CRPV), european elk papillomavirus (EEPV), rhesus monkey papillomavirus (RhPV), human papillomavirus (HPV), or a combination thereof.

[0040] In some embodiments, the human papillomavirus (HPV) is selected from the group comprising HPV1, HPV2, HPV3, HPV4, HPV5, HPV6, HPV7, HPV8, HPV9, HPV10, HPV11, HPV12, HPV13, HPV14, HPV15, HPV16, HPV17, HPV18, HPV19, HPV20, HPV21, HPV22, HPV23, HPV24, HPV25, HPV26, HPV27, HPV28, HPV29, HPV30, HPV31, HPV32, HPV33, HPV34, HPV35, HPV36, HPV37, HPV38, HPV39, HPV40, HPV41, HPV42, HPV43, HPV44, HPV45, HPV47, HPV48, HPV49, HPV50, HPV51, HPV52, HPV53, HPV54, HPV56, HPV57, HPV58, HPV59, HPV60, HPV61, HPV62, HPV63, HPV65, HPV66, HPV67, HPV68, HPV69, HPV70, HPV71, HPV72, HPV73, HPV74, HPV75, HPV76, HPV77, HPV78, HPV80, HPV81, HPV82, HPV83, HPV84, HPV85, HPV86, HPV87, HPV88, HPV89, HPV90, HPV91, HPV92, HPV93, HPV94, HPV95, HPV96, HPV97, HPV98, HPV99, HPV100, HPV101, HPV102, HPV103, HPV104, HPV105, HPV106, HPV107, HPV108, HPV109, HPV110, HPV111, HPV112, HPV113, HPV114, HPV115, HPV116, HPV117, HPV118, HPV119, HPV120, HPV121, HPV122, HPV123, HPV124, HPV125, HPV126, HPV127, HPV128, HPV129, HPV130, HPV131, HPV132, HPV133, HPV134, HPV135, HPV136, HPV137, HPV138, HPV139, HPV140, HPV141, HPV142, HPV143, HPV144, HPV145, HPV146, HPV147, HPV148, HPV149, HPV150, HPV151, HPV152, HPV153, HPV154, HPV155, HPV156, HPV157, HPV158, HPV159, HPV160, HPV161, HPV162, HPV163, HPV164, HPV165, HPV166, HPV167, HPV168, HPV169, HPV170, HPV171, HPV172, HPV173, HPV174, HPV175, HPV176, HPV177, HPV178, HPV179, HPV180, HPV181, HPV182, HPV183, HPV184, HPV185, HPV186, HPV187, HPV188, HPV189, HPV190, HPV191, HPV192, HPV193, HPV194, HPV195, HPV196, HPV197, HPV198, HPV199, HPV200, HPV201, HPV202, HPV203, HPV204, HPV205, HPV206, HPV207, HPV208, HPV209, HPV210, HPV211, HPV212, HPV213, HPV214, HPV215, HPV216, HPV217, HPV218, HPV219, HPV220, HPV221, HPV222, or a combination thereof.

[0041] In some embodiments, the LI protein is obtained or derived from a papillomavirus. In some embodiments, the LI protein is obtained or derived from a human papillomavirus.

[0042] In some embodiments, the LI protein is obtained or derived from HPV5, HPV6, HPV8, HPV11, HPV16, HPV18, HPV26, HPV30, HPV31, HPV33, HPV34, HPV35, HPV39, HPV42, HPV43, HPV44, HPV45, HPV51, HPV52, HPV53, HPV56, HPV58, HPV59, HPV66, HPV67, HPV68, HPV69, HPV70, HPV73, HPV82, HPV85, HPV97, or a combination thereof.

[0043] In some embodiments, the target peptide is an L2 protein of a papillomavirus, including its codon optimized nucleic acid sequence, or fragments, mutants, variants, comparable equivalents, or functional analogs thereof. In some embodiments, the L2 protein has an amino acid sequence of any one of SEQ ID NOs: 1130-3675, including its codon optimized nucleic acid sequence, or fragments, mutants, variants, comparable equivalents, or functional analogs thereof.

[0044] In some embodiments, the target peptide is an El protein of a papillomavirus, including its codon optimized nucleic acid sequence, or fragments, mutants, variants, comparable equivalents, or functional analogs thereof. In some embodiments, the El protein has an amino acid sequence of any one of SEQ ID NOs: 3676-4768, including its codon optimized nucleic acid sequence, or fragments, mutants, variants, comparable equivalents, or functional analogs thereof.

[0045] In some embodiments, the target peptide is an E2 protein of a papillomavirus, including its codon optimized nucleic acid sequence, or fragments, mutants, variants, comparable equivalents, or functional analogs thereof. In some embodiments, the E2 protein has an amino acid sequence of any one of SEQ ID NOs: 4769-5850, including its codon optimized nucleic acid sequence, or fragments, mutants, variants, comparable equivalents, or functional analogs thereof. In some embodiments, the target peptide is an E4 protein of a papillomavirus, including its codon optimized nucleic acid sequence, or fragments, mutants, variants, comparable equivalents, or functional analogs thereof. In some embodiments, the E4 protein has an amino acid sequence of any one of SEQ ID NOs: 5851-6415, including its codon optimized nucleic acid sequence, or fragments, mutants, variants, comparable equivalents, or functional analogs thereof.

[0046] In some embodiments, the target peptide is an E5 protein of a papillomavirus, including its codon optimized nucleic acid sequence, or fragments, mutants, variants, comparable equivalents, or functional analogs thereof. In some embodiments, the E5 protein has an amino acid sequence of any one of SEQ ID NOs: 6416-6713, including its codon optimized nucleic acid sequence, or fragments, mutants, variants, comparable equivalents, or functional analogs thereof.

[0047] In some embodiments, the target peptide is an E6 protein of a papillomavirus, including its codon optimized nucleic acid sequence, or fragments, mutants, variants, comparable equivalents, or functional analogs thereof. In some embodiments, the E6 protein has an amino acid sequence of any one of SEQ ID NOs: 6714-7420, including its codon optimized nucleic acid sequence, or fragments, mutants, variants, comparable equivalents, or functional analogs thereof.

[0048] In some embodiments, the target peptide is an E7 protein of a papillomavirus, including its codon optimized nucleic acid sequence, or fragments, mutants, variants, comparable equivalents, or functional analogs thereof. In some embodiments, the E7 protein has an amino acid sequence of any one of SEQ ID NOs: 7421-8044, including its codon optimized nucleic acid sequence, or fragments, mutants, variants, comparable equivalents, or functional analogs thereof.

[0049] In some embodiments, the target peptide is a B cell epitope of a papillomavirus, including its codon optimized nucleic acid sequence, or fragments, mutants, variants, comparable equivalents, or functional analogs thereof. In some embodiments, the B cell epitope has an amino acid sequence of any one of SEQ ID NOs: 8045-8654, including its codon optimized nucleic acid sequence, or fragments, mutants, variants, comparable equivalents, or functional analogs thereof.

[0050] In some embodiments, the target peptide is a T cell epitope of a papillomavirus, including its codon optimized nucleic acid sequence, or fragments, mutants, variants, comparable equivalents, or functional analogs thereof. In some embodiments, the T cell epitope has an amino acid sequence of any one of SEQ ID NOs: 8655-9408, including its codon optimized nucleic acid sequence, or fragments, mutants, variants, comparable equivalents, or functional analogs thereof.

[0051] In some embodiments, the target peptide has an amino acid sequence of any one of SEQ ID NOs: 1130-9408, including its codon optimized nucleic acid sequence, or fragments, mutants, variants, comparable equivalents, or functional analogs thereof.

[0052] In another aspect, provided herein is a lipid nanoparticle composition comprising a cationic lipid, a phospholipid, a sterol, a PEG-lipid, and the multisubunit nucleic acid according to any of the preceding embodiments or paragraphs. In some embodiments, the cationic lipid comprises an ionizable lipid. In some embodiments, the cationic lipid is present in an amount from 10 mol percent to 70 mol percent. In some embodiments, the phospholipid is present in an amount from 2 mol percent to 65 mol percent. In some embodiments, the sterol is present in an amount from 20 mol percent to 65 mol percent. In some embodiments, the PEG-lipid is present in an amount from 0.2 mol percent to 2.0 mol percent. In some embodiments, the lipid nanoparticle composition additionally comprises an ionizable polymer. In some embodiments, the ionizable polymer is present in an amount from 1 mol percent to 25 mol percent. In some embodiments, the ionizable polymer is selected from the group comprising a chitosan, chitosan derivatives, cellulose derivatives, a poly-L-lysine (PLL), a protamine, a polyethyleneimine, and / or their derivatives or a combination thereof. In some embodiments, the cationic lipid is represented by any one of formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), or SM-102, or ALC-0315, or a combination thereof.

[0053] In some aspects, provided herein is a vaccine comprising a lipid nanoparticle composition comprising a cationic lipid, a phospholipid, a sterol, a PEG-lipid, and the multisubunit nucleic acid according to any of the preceding embodiments or paragraphs. In some embodiments, the cationic lipid comprises an ionizable lipid. In some embodiments, the cationic lipid is present in an amount from 10 mol percent to 70 mol percent. In some embodiments, the phospholipid is present in an amount from 2 mol percent to 65 mol percent. In some embodiments, the sterol is present in an amount from 20 mol percent to 65 mol percent. In some embodiments, the PEG-lipid is present in an amount from 0.2 mol percent to 2.0 mol percent. In some embodiments, the lipid nanoparticle composition additionally comprises an ionizable polymer. In some embodiments, the ionizable polymer is present in an amount from 1 mol percent to 25 mol percent. In some embodiments, the ionizable polymer is selected from the group comprising a chitosan, chitosan derivatives, cellulose derivatives, a poly-L-lysine (PLL), a protamine, a polyethyleneimine, and / or their derivatives or a combination thereof. In some embodiments, the cationic lipid is represented by any one of formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), or SM-102, or ALC-0315, or a combination thereof.

[0054] In another aspect, provided herein is a method of treating or preventing a disease, comprising administering to a subject in need thereof, the multisubunit nucleic acid disclosed herein.

[0055] In another aspect, provided herein is a method of treating or preventing a disease, comprising administering to a subject in need thereof, the multisubunit peptide disclosed herein.

[0056] In another aspect, provided herein is a method of treating or preventing a disease, comprising administering to a subject in need thereof, the lipid nanoparticle or the lipid nanoparticle composition disclosed herein.

[0057] In another aspect, provided herein is a method of treating or preventing a disease, comprising administering to a subject in need thereof, the vaccine comprising the multisubunit nucleic acid disclosed herein.

[0058] In another aspect, provided herein is a method of treating or preventing a disease, comprising administering to a subject in need thereof, the vaccine comprising the multisubunit peptide disclosed herein.

[0059] In another aspect, provided herein is a method of treating or preventing a disease, comprising administering to a subject in need thereof, the vaccine comprising the lipid nanoparticle or the lipid nanoparticle composition disclosed herein.

[0060] In another aspect, provided herein is use of the multisubunit nucleic acid disclosed herein, in the manufacture of a medicament for the treatment or prevention of a disease in a subject.

[0061] In another aspect, provided herein is use of the vaccine comprising the multisubunit nucleic acid disclosed herein, in the manufacture of a medicament for the treatment or prevention of a disease in a subject.

[0062] In another aspect, provided herein is use of the lipid nanoparticle composition disclosed herein, in the manufacture of a medicament for the treatment or prevention of a disease in a subject. In another aspect, provided herein is use of the vaccine comprising the lipid nanoparticle or lipid nanoparticle composition disclosed herein, in the manufacture of a medicament for the treatment or prevention of a disease in a subject.

[0063] In another aspect, provided herein is a multisubunit peptide encoded by the multisubunit nucleic acid disclosed herein.

[0064] In another aspect, provided herein is a vaccine comprising a multisubunit nuclei acid sequence encoding the multisubunit peptide described herein.

[0065] In another aspect, provided herein is a multisubunit peptide, wherein the multisubunit peptide comprises either a plurality of self-assembling immunogenic peptides, or a plurality of self-assembling immunogenic peptides each further comprising one or more target peptide, or a combination thereof, wherein each self-assembling immunogenic peptide is connected to an adjacent self-assembling immunogenic peptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus or N-terminus of one or more of the self-assembling immunogenic sequences of the plurality.

[0066] In another aspect, provided herein is a multisubunit peptide comprising a plurality of self-assembling immunogenic peptides, wherein each self- assembling immunogenic peptide of the plurality is connected to an adjacent self-assembling immunogenic peptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus or N-terminus of one or more of the self-assembling immunogenic peptides of the plurality.

[0067] In another aspect, provided herein is a multisubunit peptide comprising a plurality of self-assembling immunogenic peptides, wherein each self- assembling immunogenic peptide further comprises one or more target peptide, wherein each self-assembling immunogenic peptide of the plurality is connected to an adjacent self-assembling immunogenic peptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus or N-terminus of one or more of the self-assembling immunogenic peptides of the plurality.

[0068] In another aspect, provided herein is a multisubunit peptide comprising a plurality of polypeptides, wherein some or all polypeptides of the plurality comprises either a selfassembling immunogenic peptide, or a self- assembling immunogenic peptide each further comprising one or more target peptide, or a combination thereof, wherein each polypeptide of the plurality is connected to an adjacent polypeptide of the plurality by one or more cleavage peptide, and wherein the polypeptide further comprises one or more signal peptide on the amino-terminus or N-terminus of one or more of the polypeptides of the plurality.

[0069] In another aspect, provided herein is a vaccine comprising a multisubunit peptide, wherein the multisubunit peptide comprises either a plurality of self-assembling immunogenic peptides, or a plurality self-assembling immunogenic peptides each further comprising one or more target peptide, or a combination thereof, wherein each selfassembling immunogenic peptide is connected to an adjacent self-assembling immunogenic peptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus or N-terminus of one or more of the self-assembling immunogenic peptides of the plurality.

[0070] In another aspect, provided herein is a vaccine comprising a multisubunit peptide, wherein the multisubunit peptide comprises a plurality of self-assembling immunogenic peptides, wherein each self-assembling immunogenic peptide of the plurality is connected to an adjacent self-assembling immunogenic peptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus or N-terminus of one or more of the self-assembling immunogenic peptides of the plurality.

[0071] In another aspect, provided herein is a vaccine comprising a multisubunit peptide, wherein the multisubunit peptide comprises a plurality of self-assembling immunogenic peptides, wherein each self-assembling immunogenic peptide further comprises one or more target peptide, wherein each self-assembling immunogenic peptide of the plurality is connected to an adjacent self-assembling immunogenic peptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus or N-terminus of one or more of the selfassembling immunogenic peptides of the plurality.

[0072] In another aspect, provided herein is a vaccine comprising a multisubunit peptide, wherein the multisubunit peptide comprises a plurality of polypeptides, wherein some or all polypeptides of the plurality comprises either a self-assembling immunogenic peptide, or a self-assembling immunogenic peptide each further comprising one or more target peptide, or a combination thereof, wherein each polypeptide of the plurality is connected to an adjacent polypeptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus or N-terminus of one or more of the polypeptides of the plurality.

[0073] In another aspect, provided herein is a polypeptide nanoparticle or peptide nanoparticle comprising at least 2 or up to 500 polypeptides or self-assembling immunogenic peptides respectively, disclosed herein.

[0074] In some embodiments, the self-assembling immunogenic peptides are homologous self-assembling immunogenic peptides, heterologous self-assembling immunogenic peptides, oligomeric complexes, or combination thereof. In some embodiments, the polypeptides are homologous polypeptides, heterologous polypeptides, oligomeric complexes, or a combination thereof.

[0075] In some embodiments, the peptide nanoparticle is icosahedral, helical, spherical, rod-like, or a combination thereof. In some embodiments, the polypeptide nanoparticle is icosahedral, helical, spherical, rod-like, or a combination thereof.

[0076] In some embodiments, the multisubunit nucleic acid sequence is a DNA or an RNA. In some embodiments, the multisubunit nucleic acid sequence is an mRNA. In some embodiments, the multisubunit nucleic acid sequence encodes a multisubunit peptide. In some embodiments, the multisubunit nucleic acid sequence is encapsulated or formulated in a lipid nanoparticle or lipid nanoparticle composition. In some embodiments, the multisubunit nucleic acid sequence is synthesized through one or more in vitro transcription (IVT) process. In some embodiments, the multisubunit nucleic acid sequence (for example, mRNA) is synthesized or obtained through a single in vitro transcription (IVT) process or step.

[0077] In some embodiments, the disclosure relates to a multisubunit nucleic acid sequence encoding the multisubunit peptide described herein.

[0078] In some embodiments, the disclosure relates to a vaccine comprising a multisubunit nucleic acid sequence encoding a multisubunit peptide described herein.

[0079] In some embodiments, the disclosure relates to a peptide nanoparticle or a polypeptide nanoparticle formed from the self-assembly of two or more self-assembling immunogenic peptides or the polypeptides, respectively. In some embodiments, the selfassembling immunogenic peptides or the polypeptides in the peptide nanoparticle or polypeptide nanoparticle, respectively, may also have some residues (amino acids) of the cleavage peptide. In some embodiments, the polypeptide nanoparticle comprises homologous polypeptides, heterologous polypeptides, oligomeric complexes, or a combination thereof.

[0080] In some embodiments, the peptide nanoparticle comprises homologous selfassembling immunogenic peptides, heterologous self-assembling immunogenic peptides, oligomeric complexes, or a combination thereof.

[0081] In some aspects, provided herein is the multisubunit nucleic acid sequence described herein, encapsulated or formulated in a lipid nanoparticle composition. In some aspects, the lipid nanoparticle composition comprises a cationic lipid, a phospholipid, a sterol, a PEG lipid and the multisubunit nucleic acid sequence described herein. In some embodiments, the cationic lipid is represented by any one of formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), or SM-102, or ALC-0315, or a combination thereof.

[0082] In some aspects, provided herein is a vaccine comprising the multisubunit nucleic acid sequence described herein, encapsulated or formulated in a lipid nanoparticle composition. In some aspects, the lipid nanoparticle composition comprises a cationic lipid, a phospholipid, a sterol, a PEG lipid and the multisubunit nucleic acid sequence described herein. In some embodiments, the cationic lipid is represented by any one of formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), or SM-102, or ALC-0315, or a combination thereof.

[0083] In some aspects, the lipid nanoparticle composition comprises an ionizable polymer, a cationic lipid, a phospholipid, a sterol, a PEG-lipid and the multisubunit nucleic acid sequence described herein. In some embodiments, the cationic lipid is represented by any one of formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), or SM-102, or ALC-0315, or a combination thereof.

[0084] In some aspects, provided herein is a method of treating or preventing a disease, comprising administering to a subject in need thereof the multisubunit nucleic acid sequence as described herein.

[0085] In some aspects, provided herein is a method of treating or preventing a disease comprising administering to a subject in need thereof a vaccine comprising the multisubunit nucleic acid sequence as described herein.

[0086] In some aspects, provided herein is a method of treating or preventing a disease, comprising administering to a subject in need thereof a lipid nanoparticle composition comprising a cationic lipid, a phospholipid, a sterol, a PEG-lipid and the multisubunit nucleic acid sequence as described herein. In some embodiments, the cationic lipid is represented by any one of formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), or SM-102, or ALC-0315, or a combination thereof.

[0087] In some aspects, provided herein is a method of treating or preventing a disease, comprising administering to a subject in need thereof a lipid nanoparticle composition comprising an ionizable polymer, a cationic lipid, a phospholipid, a sterol, a PEG-lipid and the multisubunit nucleic acid sequence as described herein. In some embodiments, the cationic lipid is represented by any one of formula (I), formula (II), formula (III), formula

[0088] (IV), formula (V), formula (VI), formula (VII), formula (VIII), or SM-102, or ALC-0315, or a combination thereof.

[0089] In some aspects, the disclosure relates to use of a lipid nanoparticle composition comprising a cationic lipid, a phospholipid, a sterol, a PEG-lipid, and the multisubunit nucleic acid sequence as described herein, in the manufacture of a medicament for the treatment or prevention of a disease in a subject. In some embodiments, the cationic lipid is represented by any one of formula (I), formula (II), formula (III), formula (IV), formula

[0090] (V), formula (VI), formula (VII), formula (VIII), or SM-102, or ALC-0315, or a combination thereof.

[0091] In some aspects, the disclosure relates to use of a vaccine comprising a lipid nanoparticle composition, wherein the lipid nanoparticle composition comprises a cationic lipid, a phospholipid, a sterol, a PEG-lipid, and the multisubunit nucleic acid sequence as described herein, in the manufacture of a medicament for the treatment or prevention of a disease in a subject. In some embodiments, the cationic lipid is represented by any one of formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), or SM-102, or ALC-0315, or a combination thereof.

[0092] In some embodiments, the target sequence is obtained or derived from L2 protein, El protein, E2 protein, E4 protein, E5 protein, E6 protein, E7 protein, B cell epitope, T cell epitope, or a combination thereof of a papillomavirus, including the codon optimized sequences, fragments, variants, mutants, comparable equivalents, or functional analogs of such target sequences. In some embodiments, the target sequence is modified or unmodified.

[0093] In some embodiments, the target sequence encodes a target peptide obtained or derived from L2 protein, El protein, E2 protein, E4 protein, E5 protein, E6 protein, E7 protein, B cell epitope, T cell epitope, or a combination thereof of a papillomavirus. In some embodiments, the target sequence or target peptide is modified or unmodified. In some embodiments, the target peptide regulates or modulates cellular functions. In some embodiments, the target peptide has immunostimulatory or immunomodulatory effect. In some embodiments, the target peptide may have regulatory effect. In some embodiments, the target peptide has suppressor effect.

[0094] In some embodiments, the self-assembling immunogenic sequence encodes a selfassembling immunogenic peptide. In some embodiments, the self-assembling immunogenic peptide is obtained or derived from LI protein of a papillomavirus, including their fragments, mutants, variants, comparable equivalents, or functional analogs thereof. In some embodiments, the LI protein comprises one or more mutations. In some embodiments, the mutation in the LI protein is a substitution replacing amino acid cysteine (C) with amino acid alanine (A), at the amino acid position 175 (i.e., C175A), compared to SEQ ID NO: 712. In some embodiments, the mutation in the LI protein is a substitution replacing amino acid cysteine (C) with amino acid alanine (A), at the amino acid position 428 (i.e., C428A), compared to SEQ ID NO: 712. In some embodiments, the mutation in the LI protein is selected from C175A and / or C428A compared to SEQ ID NO: 712.

[0095] In some embodiments, the linker sequence encodes a linker peptide. In some embodiments, the linker peptide connects the target peptide with the self-assembling immunogenic peptide. In some embodiments, the linker peptide connects two signal peptides. In some embodiments, the linker peptide connects two cleavage peptides. The linker peptide may be an amino acid linker, a foldon, a scaffold, or a combination thereof.

[0096] In some embodiments, the cleavage sequence encodes a cleavage peptide. In some embodiments, the cleavage peptide comprises one or more cleavage peptides. In some embodiments, the cleavage peptide connects one self-assembling immunogenic peptide with another self-assembling immunogenic peptide, for example, adjacent self- assembling immunogenic peptide. In some embodiments, the cleavage peptide connects one polypeptide with another polypeptide, for example, adjacent polypeptide. The cleavage peptide carries a cleavage site. In some embodiments, the cleavage peptide carries one or more cleavage sites. In some embodiments, the cleavage peptide facilitates the action of cellular proteases to cleave the multisubunit peptide into individual self- assembling immunogenic peptides or polypeptides. In some embodiments, the cleavage peptide self cleaves (for example, without bond formation during translation of multisubunit nucleic acid) into individual self-assembling immunogenic peptides or polypeptides. In some embodiments, the cleavage peptide comprises two or more cleavage peptides (for example, cleavage peptide- 1, cleavage peptide-2 and so on), optionally connected via a linker peptide. In some embodiments, the cleavage peptide self cleaves into individual selfassembling immunogenic peptides or polypeptides, or is cleaved by the action of cellular proteases. In some embodiments, the cleavage peptide is a substrate for cellular proteases. In some embodiments, the cleavage peptide is a substrate for golgi specific proteases. In some embodiments, the cleavage peptide is a self cleaving peptide. In some embodiments, the cleavage peptide comprises two or more cleavage peptides (for example, cleavage peptide- 1, cleavage peptide-2 and so on), optionally linked by a linker peptide, wherein one cleavage peptide is a substrate for cellular proteases and the other cleavage peptide is a self cleaving peptide.

[0097] In some embodiments, the signal sequence encodes a signal peptide. The signal peptide is present upstream (amino-terminus) of one or more self-assembling immunogenic peptides or polypeptides in a multisubunit peptide. In some embodiments, the signal peptide is present upstream (amino-terminus) of the first self-assembling immunogenic peptide or the first polypeptide. In some embodiments, the signal peptide is present upstream (amino-terminus) of some self-assembling immunogenic peptides or some polypeptides. In some embodiments, the signal peptide is present upstream (aminoterminus) of all self-assembling immunogenic peptides or all polypeptides. In some embodiments, the signal peptide transports the multisubunit peptide to cell organelles. In some embodiments, the signal peptide transports the multisubunit peptide to golgi body or golgi apparatus. In some embodiments, the signal peptide is a golgi targeting signal peptide.

[0098] In some aspects, the present disclosure also includes a method of transforming a cell with the multisubunit nucleic acid sequence as described herein.

[0099] BRIEF DESCRIPTION OF THE DRAWINGS

[0100] Figure 1 - shows representative schematic illustration of multisubunit nucleic acid sequence wherein each self-assembling immunogenic sequence (SAIS) is connected to an adjacent self-assembling immunogenic sequence (SAIS) by a cleavage sequence (CS) such that between any two self- assembling immunogenic sequence there is present a cleavage sequence. The multisubunit nucleic acid sequence has a signal sequence (SS) upstream of the first self-assembling immunogenic sequence. The self-assembling immunogenic sequence can either be self-assembling immunogenic sequence or self-assembling immunogenic sequence further comprising (i.e., into which is inserted) one or more target sequence (TS). The letter ‘n’ in figure 1 represents any number between 1 to 98. The multisubunit nucleic acid sequence may additionally have 5’ cap and 3’ poly(A) tail. This multisubunit nucleic acid sequence encodes corresponding multisubunit peptide depicted in Figure 2.

[0101] Figure 2 - shows representative schematic illustration of multisubunit peptide wherein each self-assembling immunogenic peptide (SAIP) is connected to an adjacent self-assembling immunogenic peptide (SAIP) by a cleavage peptide (CP) such that between any two self-assembling immunogenic peptide there is present a cleavage peptide. The multisubunit peptide has a signal peptide (SP) on the N-terminus of the first selfassembling immunogenic peptide. The self-assembling immunogenic peptide can either be self-assembling immunogenic peptide or self-assembling immunogenic peptide further comprising (i.e., into which is inserted) one or more target peptide (TP). The letter ‘n’ in figure 1 represents any number between 1 to 98.

[0102] Figure 3 - shows representative schematic illustration of multisubunit nucleic acid sequence wherein each polynucleotide sequence (PS) comprises either a self- assembling immunogenic sequence (SAIS) or self-assembling immunogenic sequence further comprising (i.e., into which is inserted) one or more target sequence (TS) or a combination thereof. Multiple polynucleotide sequences are connected through a cleavage sequence (CS) such that between any two polynucleotide sequences there is present a cleavage sequence. The multisubunit nucleic acid sequence has a signal sequence (SS) upstream of the first polynucleotide sequence. The letter ‘n’ in figure 3 represents any number between 1 to 98. The multisubunit nucleic acid sequence may additionally have 5’ cap and 3’ poly(A) tail. This multisubunit nucleic acid sequence encodes corresponding multisubunit peptide depicted in Figure 4.

[0103] Figure 4 - shows representative schematic illustration of multisubunit peptide wherein each polypeptide (PP) comprises either a self-assembling immunogenic peptide (SAIP) or self-assembling immunogenic peptide further comprising (i.e., into which is inserted) one or more target peptide (TP) or a combination thereof. Multiple polypeptides are connected through a cleavage peptide (CP) such that between any two polypeptides there is present a cleavage peptide. The multisubunit peptide has a signal peptide (SP) on the N-terminus of the first polypeptide. The letter ‘n’ in figure 4 represents any number between 1 to 98.

[0104] Figures 5, 6, and 7 - shows representative schematic illustration of different manifestations of multisubunit nucleic acid of figure 1. SS, SAIS, CS, TS, and the letter ‘n’ represents the same elements and the number respectively as in Figure 1, with the caveat that the sum of the number represented by both the ns in Figure 7 is not more than 99.

[0105] Figures 8, 9, and 10 - shows representative schematic illustration of different manifestations of the multisubunit peptide of Figure 2, encoded by the multisubunit nucleic acid of figure 1. SP, SAIP, CP, TP and the letter ‘n’ represents the same elements and the number respectively as in Figure 2, with the caveat that the sum of the number represented by both the ns in Figure 10 is not more than 99.

[0106] DESCRIPTION

[0107] The present disclosure relates to multisubunit nucleic acid sequences, multisubunit peptides, and peptide nanoparticles or polypeptide nanoparticles and their compositions for vaccine and therapeutic purpose against papillomavirus.

[0108] Unless defined otherwise, technical, and scientific terms used herein have the same meaning as commonly understood by one of person skill in the art. Some of the terms are defined briefly here below; the definitions should not be construed in a limiting sense.

[0109] The singular forms “a”, “an” and “the” as used in the specification also include plural aspects unless the context dictates otherwise. Similarly, any singular term used in the specification also mean plural or vice versa unless the context dictates otherwise. As used herein in the claim(s), when used in conjunction with the word “comprising”, the words “a” or “an” may mean one or more than one. As used herein “another” may mean at least a second or more.

[0110] It must be noted that the words “comprising” or any of its form such as “comprise” or “comprises”, “having” or any of its forms such as “have” or “has”, “including” or any of its forms such as “include” or “includes”, or “containing” or any of its forms such as “contain” or “contains” are open-ended and do not exclude additional unrecited elements or method steps. Wherever any quantity or range is stated one skilled in the art will recognize that quantity or range within 10 or 20 percent of the stated values can also be expected to be appropriate and reasonable and included within the scope of the invention.

[0111] Unless otherwise defined herein, scientific, and technical terms used in connection with the present invention shall have the meanings that are commonly understood by those of ordinary skilled in the art. Generally, nomenclatures used in connection with, and techniques of, cell and tissue culture, molecular biology, immunology, microbiology, protein, adjuvant, pharmaceutical biotechnology, and biopharmaceutical manufacturing described herein are those well known and commonly used in the art. The methods and techniques of the present invention are generally performed according to conventional methods well known in the art and as described in various general and more specific references that are cited and discussed throughout the present specification.

[0112] The term “composition”, “formulation”, “lipid nanoparticle composition”, or “lipid nanoparticle” has been used interchangeably to mean a nanoparticle, nanostructure, vesicle, liposome, composition or formulation comprising one or more lipid components (for example, a cationic lipid, a phospholipid, a sterol, and a PEG-lipid), and / or an ionizable polymer component. In some embodiments, the lipid nanoparticle comprises a cationic lipid, a phospholipid, a sterol, a PEG-lipid, and a multisubunit nucleic acid. In some embodiments, the cationic lipid is represented by any one of formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), or SM-102, or ALC-0315, or a combination thereof. In some embodiments, the lipid nanoparticle comprises one or more lipid components, an ionizable polymer component, and a multisubunit nucleic acid. In some embodiments, the multisubunit nucleic acid associated with the lipid nanoparticle is a DNA, an mRNA, a micro RNA, a small interfering RNA, a small nucleolar RNA, a small nuclear RNA, a long non-coding RNA or a combination thereof. In some embodiments, the lipid nanoparticle composition contains one or more pharmaceutical carriers or excipients, such as but not limited to, buffering agents, stabilizers, tonicity modifiers, surfactants, chelating agents, salts, anti-oxidants, diluents, and / or preservatives or a combination thereof. The term lipid nanoparticle also denotes lipid nanoparticles that are devoid of any encapsulated multisubunit nucleic acid (empty lipid nanoparticles or ghost lipid nanoparticles). In some embodiments, the lipid nanoparticle composition comprises lipid nanoparticles with encapsulated multisubunit nucleic acid as well as empty lipid nanoparticles. The term “therapeutic”, “therapeutic agent”, “prophylactic”, “prophylactic agent”, or “drug” has been used interchangeably to mean a compound (such as multisubunit nucleic acid sequence) or composition (such as a lipid nanoparticle composition described herein) having a biological effect or a combination of biological effects that prevents, inhibits, eliminates or prevents the progression of a disease or other aberrant biological processes in a subject, for example, an animal or human.

[0113] The term “preventing” is art-recognized, and when used in relation to a condition, such as an infection is well understood in the art, and includes administration of a composition, which reduces the frequency or severity, or delays the onset, of one or more symptoms of the medical condition in a subject relative to a subject who does not receive the composition. Thus, the prevention of a condition, such as an infection, includes, for example, the reduction of the frequency or severity of one or more symptoms of the medical condition in a population of patients receiving a therapy relative to a control population that did not receive the therapy, e.g., by a statistically and / or clinically significant amount. Similarly, the prevention of an infection includes reducing the likelihood that a patient receiving a therapy will develop the infection or related symptoms, relative to a patient who does not receive the therapy.

[0114] The term “molar percent”, “mol percent”, “molar %”, or “mol %” have been used interchangeably to mean number of moles of a component expressed as percentage relative to total moles of all lipid components present in the lipid nanoparticle compositions described herein. For example, 50 mol % cationic lipid means, 50 mol % of cationic lipid is present in the lipid nanoparticle composition and other lipid components together constitute remaining 50 mol % such that the total amount of all the lipid components constitute 100 mol %. In some embodiments, mol % also denotes to mean number of moles of a component expressed as percentage relative to total moles of all lipid components (such as cationic lipid, phospholipid, sterol and PEG-lipid) and ionizable polymer component(s) present in the lipid nanoparticle composition described herein. For example, 50 mol % of cationic lipid means, 50 mol % of cationic lipid is present in the lipid nanoparticle composition and other lipids components and ionizable polymer components together constitute the remaining 50 mol % such that the total amount of all the lipid components and ionizable polymer components constitute 100 mol %.

[0115] The term “N / P ratio”, “N:P ratio”, “lipid to nucleic acid ratio”, or “cationic lipid to nucleic acid ratio” have been used interchangeably herein and means the ratio (molar ratio) of the positive charges in the cationic lipid relative to the negative charges in the nucleic acid (such as the multisubunit nucleic acid sequences disclosed herein) in a lipid nanoparticle. In some embodiments, the N / P ratio refers to the ratio of protonable nitrogen present in the cationic lipid relative to the phosphate present in the nucleic acid in a lipid nanoparticle. In some embodiments, the N / P ratio is between 1 to 18 (i.e., 1 :1 to 18:1). For example, a N / P ratio of 18 refers to the presence of 18 protonable nitrogen of the cationic lipid relative to 1 phosphate of the nucleic acid in a lipid nanoparticle.

[0116] The terms “antibody” and “antibodies” have been used interchangeably herein and means any antibody or antibody fragment (whether produced naturally or recombinantly) which retains antigen binding activity. This includes a monoclonal or polyclonal antibody, a single chain antibody, a Fab fragment of a monoclonal or polyclonal antibody, a chimeric antibody, a humanized antibody, a human antibody, a bispecific antibody, a multispecific antibody, or a nanobody.

[0117] The term “buffer” as used herein means those agents that maintains the pH of a solution in a desired range.

[0118] The term “cell” as used herein means a single cell or a population of cells or plurality of cells.

[0119] The term “biologically effective amount” or “therapeutically effective amount” as used herein means an amount of an agent, for example, a therapeutic, drug, therapeutic agent, prophylactic agent, diagnostic agent, composition, etc., that is sufficient, when administered to a subject suffering from or susceptible to an infection, disease, disorder, and / or condition, to treat, prevent, diagnose, improve symptoms of, and / or delay the onset of the infection, disease, disorder, and / or condition. A therapeutically effective amount herein may vary according to factors such as the disease state, age, sex, and weight of the patient.

[0120] As used herein, the term “treating” or “treatment” includes reducing, arresting, or reversing the symptoms, clinical signs, or underlying pathology of a condition to stabilize or improve a subject’s condition or to reduce the likelihood that the subject’s condition will worsen as much as if the subject did not receive the treatment. Treatment may be administered to a subject who does not exhibit signs of a disease and / or exhibits only early signs of the disease for the purpose of decreasing the risk of developing pathology associated with the disease. The term “subject” as used herein refers to an animal or human, for example a living mammal and may be interchangeably used with the term “patient”. Examples of mammals include, but are not limited to, any member of the mammalian class: humans, non-human primates such as chimpanzees, and other apes and monkey species; farm animals such as cattle, horses, sheep, goats, swine; domestic animals such as rabbits, dogs, and cats; laboratory animals including rodents, such as rats, mice, ferrets, and guinea pigs, and the like. The term does not denote a particular age or gender.

[0121] As used herein, an individual “at risk” of developing a particular disease, disorder, or condition may or may not have detectable disease or symptoms of disease, and may or may not have displayed detectable disease or symptoms of disease prior to the treatment methods described herein. “At risk” denotes that an individual has one or more risk factors, which are measurable parameters that correlate with development of a particular disease, disorder, or condition, as known in the art. An individual having one or more of these risk factors has a higher probability of developing a particular disease, disorder, or condition than an individual without one or more of these risk factors.

[0122] The term “disease” as used herein, means an interruption, cessation, or disorder of body function, system, or organ. Non limiting examples of disease include malignant diseases, autoimmune diseases, inherited diseases, metabolic disorders, or infectious diseases.

[0123] The term “vaccine” as used herein means a substance or composition comprising an antigen or immunogen for eliciting an immune response in a subject against the antigen or the immunogen. The term vaccine is also understood to mean a substance or composition comprising an antigen or immunogen that activates or stimulates an immune cell. In some cases, the antigen or immunogen is a peptide, a protein, a polysaccharide, or a combination thereof. In some cases, the antigen or immunogen is encoded by a nucleic acid, for example, a DNA, an RNA, or an mRNA. In some embodiments, the vaccine comprises a nucleic acid that encodes an antigen or an immunogen. In some embodiments, the vaccine comprises a multisubunit nucleic acid as described herein.

[0124] As used herein, administration “conjointly” with another compound or composition includes simultaneous administration and / or administration at different times. Conjoint administration also encompasses administration as a co-formulation or administration as separate compositions, including at different dosing frequencies or intervals, and using the same route of administration or different routes of administration. The term “multisubunit nucleic acid sequence” or “multisubunit nucleic acid” have been used interchangeably and means two or more self- assembling immunogenic sequences or polynucleotide sequences wherein the self-assembling immunogenic sequence or polynucleotide sequence is connected to another self-assembling immunogenic sequence or polynucleotide sequence, respectively, by a cleavage sequence, wherein the multisubunit nucleic acid sequence includes one or more signal sequence upstream of one or more self-assembling immunogenic sequences or polynucleotide sequences. In some embodiments, the self-assembling immunogenic sequence includes a target sequence inserted into it. In some embodiments, the self-assembling immunogenic sequence comprises a linker sequence on one or both sides (5’ [5-prime] or 3’ [3-prime]) of the target sequence. In some embodiments, the signal sequence is present upstream of the first self-assembling immunogenic sequence or the first polynucleotide sequence. In some embodiments, the signal sequence is present upstream of some self-assembling immunogenic sequences or some polynucleotide sequences. In some embodiments, the signal sequence is present upstream of each of the self-assembling immunogenic sequences or each of the polynucleotide sequences. In some embodiments, the selfassembling immunogenic sequences comprises a target sequence. In some embodiments, the self-assembling immunogenic sequence comprises a linker sequence on one or both sides of the target sequence. As illustrated in figure 1, 3, 5, 6, & 7 multisubunit nucleic acid sequence comprises multiple repeats of self-assembling immunogenic sequences or polynucleotide sequences, such that total number of self-assembling immunogenic sequences or polynucleotide sequences in a multisubunit nucleic acid sequence are not more than 100. In some embodiments, the signal sequence is present upstream of each of some or all of the self-assembling immunogenic sequence or each of some or all of the polynucleotide sequences. In some embodiments, the multisubunit nucleic acid sequence is obtained or synthesized through single in vitro transcription (IVT) process or step. The multisubunit nucleic acid sequence encodes a multisubunit peptide.

[0125] The terms “multisubunit peptide” as used herein means two or more selfassembling immunogenic peptides or polypeptides wherein the self-assembling immunogenic peptide or polypeptide is connected to another self-assembling immunogenic peptide or polypeptide, respectively, by a cleavage peptide, wherein the multisubunit peptide includes one or more signal peptide on the N-terminus of one or more selfassembling immunogenic peptides or polypeptides. In some embodiments, the self- assembling immunogenic peptide includes a target peptide inserted into it. In some embodiments, the self-assembling immunogenic peptide comprises a linker peptide on one or both sides (i.e., on the C-terminus or N-terminus or both sides) of the target peptide. In some embodiments, the signal peptide is present on the N-terminus of the first selfassembling immunogenic peptide or the first polypeptide. In some embodiments, the signal peptide is present on the N-terminus of some self-assembling immunogenic peptides or some polypeptides. In some embodiments, the signal peptide is present on the N-terminus of each of the self-assembling immunogenic peptides or each of the polypeptides. In some embodiments, the self-assembling immunogenic peptide comprises a target peptide. In some embodiments, the self-assembling immunogenic peptide comprises a linker peptide on one or both side of the target peptide. As illustrated in figures 2, 4, 8, 9, & 10 multisubunit peptide comprises multiple repeats of self-assembling immunogenic peptides or polypeptides, such that total number of self-assembling immunogenic peptides or polypeptides in a multisubunit peptide are not more than 100. In some embodiments, the signal peptide is present on the N-terminus of each of some or all of the self-assembling immunogenic peptides or each of some or all of the polypeptides. The multisubunit peptide may comprise homologous self-assembling immunogenic peptides or heterologous selfassembling immunogenic peptides. In some embodiments, the multisubunit peptide may comprise homologous polypeptides or heterologous polypeptides.

[0126] The term “polynucleotide sequence” as used herein means a sequence of nucleotides that encodes a polypeptide.

[0127] The terms “protein” or “peptide” have been used interchangeably herein and mean a polymer of amino acids linked through peptide bonds, but do not imply any specific length. The term also includes fusion proteins, muteins, analogs or modified forms.

[0128] The term “polypeptide” as used herein means a sequence of amino acids that comprises either a self-assembling immunogenic peptide or a self-assembling immunogenic peptide further comprising one or more target peptide. In some embodiments, the target peptide is obtained or derived from L2 protein, El protein, E2 protein, E4 protein, E5 protein, E6 protein, E7 protein, B cell epitope, T cell epitope, or a combination thereof of a papillomavirus. In some embodiments, the self-assembling immunogenic peptide includes a target peptide inserted into it. In some embodiments, the polypeptide or the self-assembling immunogenic peptide may have some residues (amino acids) of cleavage peptide. In some embodiments, the polypeptide or the self-assembling immunogenic peptide comprises a signal peptide.

[0129] The term “target sequence” as used herein means a sequence of nucleotides that encodes a target peptide obtained or derived from L2 protein, El protein, E2 protein, E4 protein, E5 protein, E6 protein, E7 protein, B cell epitope, T cell epitope, or a combination thereof of a papillomavirus.

[0130] The term “target peptide” as used herein means a sequence of amino acids that exerts a biological effect or a combination of biological effect that prevents, inhibits, eliminates, or prevents the progression of a disease or other aberrant biological process in a subject. In some embodiments, the target peptide is obtained or derived from L2 protein, El protein, E2 protein, E4 protein, E5 protein, E6 protein, E7 protein, B cell epitope, T cell epitope, or a combination thereof of a papillomavirus. In some embodiments, the target peptide in two or more self-assembling immunogenic peptides or polypeptides are identical i.e., homologous self-assembling immunogenic peptides or homologous polypeptides, respectively. In some other embodiments, the target peptides in two or more self-assembling immunogenic peptides or polypeptides are different i.e., heterologous selfassembling immunogenic peptides or heterologous polypeptides, respectively.

[0131] The term “signal sequence” as used herein means a sequence of nucleotides that encodes a signal peptide.

[0132] The term “signal peptide” as used herein means a sequence of amino acids that transports the multisubunit peptide to specific cell organelles. In some embodiments the signal peptide transports the multisubunit peptide to golgi apparatus or golgi body. The signal peptide is present on the N-terminus (amino-terminus) of one or more selfassembling immunogenic peptides or polypeptides. In some embodiments, the signal peptide is present on the N-terminus of one or more self-assembling immunogenic peptides. In some embodiments, the signal peptide is present on the N-terminus of one or more polypeptides. In some embodiments, the signal peptide is present on the N-terminus of some or all self-assembling immunogenic peptides or polypeptides. In some embodiments, the signal peptide is present on the N-terminus of some or all selfassembling immunogenic peptides. In some embodiments, the signal peptide is present on the N-terminus of some or all polypeptides. In some embodiments, the signal peptide is present on the N-terminus of the first self-assembling immunogenic peptide or the first polypeptide. In some embodiments, the signal peptide is present on the N-terminus of the first self-assembling immunogenic peptide. In some embodiments, the signal peptide is present on the N-terminus of the first polypeptide. In some embodiments, the signal peptide is present on the N-terminus of some self-assembling immunogenic peptides. In some embodiments, the signal peptide is present on the N-terminus of some polypeptides. In some embodiments, the signal peptide is present on the N-terminus of all selfassembling immunogenic peptides. In some embodiments, the signal peptide is present on the N-terminus of all polypeptides. In some embodiments, the signal peptide is encoded by one or more signal sequences. In some embodiments, the signal peptide is a golgi targeting signal peptide.

[0133] The term “cleavage sequence” as used herein means a sequence of nucleotides that encodes a cleavage peptide.

[0134] The term “cleavage peptide” as used herein means a sequence of amino acids that facilitates the action of cellular proteases to cleave the multisubunit peptide into individual self-assembling immunogenic peptides or polypeptides or self cleaves (for example, without bond formation during translation of multisubunit nucleic acid) into individual self-assembling immunogenic peptides or polypeptides respectively. The cleavage peptide is present between any two self-assembling immunogenic peptides or any two polypeptides. It connects one self-assembling immunogenic peptide or polypeptide with another self-assembling immunogenic peptide or polypeptide respectively, for example, adjacent self-assembling immunogenic peptide or polypeptide. In some embodiments, the cleavage peptide is present between any two self-assembling immunogenic peptides. In some embodiments, the cleavage peptide is present between any two polypeptides. In some embodiments, the cleavage peptide connects one self-assembling immunogenic peptide with another self-assembling immunogenic peptide, for example, adjacent selfassembling immunogenic peptide. In some embodiments, the cleavage peptide connects one polypeptide with another polypeptide, for example, adjacent polypeptide. The cleavage peptide carries one or more cleavage sites. In some embodiments, the cleavage peptide is a substrate for proteases. In some embodiments, cleavage peptide undergoes self cleavage to result in individual self-assembling immunogenic peptides or polypeptides. In some embodiments, the cleavage peptide undergoes self cleavage to result in individual self-assembling immunogenic peptides. In some embodiments, the cleavage peptide undergoes self cleavage to result in individual polypeptides. In some embodiments, the cleavage peptide is a substrate for golgi specific proteases. In some embodiments, the cleavage peptide comprises one or more cleavage peptides, for example, cleavage peptide- 1 , cleavage peptide-2 and so on. In some embodiments, the cleavage peptide optionally comprises a linker peptide between two cleavage peptides. In some embodiments, the cleavage peptide self cleaves into individual self-assembling immunogenic peptides or polypeptides respectively or is cleaved by the action of cellular proteases. In some embodiments, the cleavage peptide is a substrate for cellular proteases. In some embodiments, the cleavage peptide is a substrate for golgi specific proteases. In some embodiments, the cleavage peptide is a self cleaving peptide. In some embodiments, the cleavage peptide comprises two or more cleavage peptides (for example, cleavage peptide - 1, cleavage peptide-2 and so on), optionally linked by a linker peptide, wherein one cleavage peptide is a substrate for cellular proteases and the other cleavage peptide is a self cleaving peptide.

[0135] The term “linker sequence” as used herein means a sequence of nucleotides that encodes a linker peptide.

[0136] The term “linker peptide” or “peptide linker” have been used interchangeably to mean a sequence of amino acids that may be present on one or both sides of the target peptide. In some embodiments, the linker peptide connects two cleavage peptides. In some embodiments, the linker peptide is an amino acid linker, a foldon, a scaffold or a combination thereof. In some embodiments, the linker peptide is an amino acid linker. In some embodiments, the linker peptide is a foldon. In some embodiments, the linker peptide is a scaffold. In some embodiments, the linker peptide comprises an amino acid linker and a foldon. In some embodiments, the linker peptide comprises an amino acid linker and a scaffold. In some embodiments, the linker peptide comprises a foldon and a scaffold. In some embodiments, the linker peptide comprises an amino acid linker, a foldon, and a scaffold.

[0137] The term “amino acid linker sequence” as used herein means a sequence of nucleotides that encodes an amino acid linker.

[0138] The term “amino acid linker” as used herein means a sequence of amino acids that provides structural integrity to self- assembling immunogenic peptide or polypeptide carrying the target peptide such that the self-assembling immunogenic peptide or polypeptide remain, as far as possible, in their native or stable conformation. In some embodiments, amino acid linker also helps in orientation of a polypeptide such that the domains or epitopes on the target peptide are exposed or displayed for interaction or communication with cells or biomolecules or immune system in the absence of foldon or scaffold. In some embodiments, the amino acid linker connects two cleavage peptides. Some of the non-limiting examples of amino acid linkers includes, glycine serine linker, glycine proline linker, glycine threonine linker, alanine serine linker, any combination of two amino acids, or a combination thereof. In some embodiments, amino acid linker is about 2-49 amino acid long.

[0139] The term “glycine serine linker sequence” as used herein means a sequence of nucleotides that encodes a glycine serine linker.

[0140] The term “glycine serine linker” as used herein means a sequence of amino acid comprising one or more glycine (G) and serine (S) in any combinations without any preference of order or limitation on number of appearances of either glycine or serine. In some embodiments, the glycine serine linker is few amino acids in length to several amino acids in length.

[0141] The term “foldon sequence” as used herein means a sequence of nucleotides that encodes a foldon.

[0142] The term “foldon” as used herein means a sequence of amino acids that enables two or more homologous self-assembling immunogenic peptides or homologous polypeptides to organise to form their respective oligomeric complex. In some embodiments, the foldon also helps in orientation of a self-assembling immunogenic peptide or polypeptide such that the domains or epitopes on the target peptide are exposed or displayed for interaction or communication with cells or biomolecules or immune system.

[0143] The term “scaffold sequence” as used herein means a sequence of nucleotides that encodes a scaffold.

[0144] The term “scaffold” as used herein means a sequence of amino acids that provides structural and / or functional integrity or support to the target peptide and helps in orientation of target peptide such that the domains or epitopes of the target peptide are exposed or displayed for interaction or communication with cells or biomolecules or immune system.

[0145] The term “oligomeric complex” as used herein means a complex formed by two or more homologous self-assembling immunogenic peptides or two or more homologous polypeptides respectively. In some embodiments, the oligomeric complex has at least two homologous self-assembling immunogenic peptides, at least three homologous self- assembling immunogenic peptides, at least four homologous self-assembling immunogenic peptides, at least five homologous self-assembling immunogenic peptides, at least six homologous self-assembling immunogenic peptides and so on. In some embodiments, the oligomeric complex has at least two homologous polypeptides, at least three homologous polypeptides, at least four homologous polypeptides, at least five homologous polypeptides, or at least six homologous polypeptides and so on.

[0146] The term “self-assembling immunogenic sequence” as used herein means a sequence of nucleotides that encodes a self- assembling immunogenic peptide.

[0147] The term “self-assembling immunogenic peptide” as used herein means a sequence of amino acids that enables the self-assembling immunogenic peptides to self-assemble into peptide nanoparticle or enables the polypeptides to self-assemble into polypeptide nanoparticle. In some embodiments, the self-assembling immunogenic peptide is also capable of eliciting an immune response. In some embodiments, the self-assembling immunogenic peptide is obtained or derived from LI protein of a papillomavirus. In some embodiments, the self-assembling immunogenic peptide further comprises one or more target peptide (i.e., includes a target peptide inserted into it).

[0148] The term “self-assembly” or “self-assemble” or “self-assembling” has been used interchangeably to means the ability of self-assembling immunogenic peptides or polypeptides to undergo multimerization to form a peptide nanoparticle or polypeptide nanoparticle respectively. In some embodiments, the peptide nanoparticle or polypeptide nanoparticle has at least two self-assembling immunogenic peptides or polypeptides (dimer or 2-mer), at least three self-assembling immunogenic peptides or polypeptides (trimer or 3-mer), at least four self-assembling immunogenic peptides or polypeptides (tetramer or 4-mer), at least five self-assembling immunogenic peptides or polypeptides (pentamer or 5-mer), at least six self-assembling immunogenic peptides or polypeptides (hexamer or 6-mer), at least seven self-assembling immunogenic peptides or polypeptides (heptamer or 7-mer), at least eight self- assembling immunogenic peptides or polypeptides (octamer or 8-mer), and so on, respectively. In some embodiments, the peptide nanoparticle or polypeptide nanoparticle is up to 500-mers. In some embodiments, hydrogen bonds, disulfide bonds, hydrophobic interactions, electrostatic interactions, and / or Van der Waals forces combine to maintain self-assembled structure.

[0149] The term “multimerization” as used herein means association of two or more units of homologous self-assembling immunogenic peptides, or heterologous self-assembling immunogenic peptides, or oligomeric complexes, or their combination. The term multimerization is also referred to the association of two or more units of homologous polypeptides or heterologous polypeptides, or oligomeric complexes or their combination.

[0150] The term “peptide nanoparticle” as used herein means a nanoparticle formed by self-assembly of self-assembling immunogenic peptides. The self-assembling immunogenic peptides may or may not include the target peptide inserted into them. In some embodiments, the peptide nanoparticle comprises two or more homologous selfassembling immunogenic peptides, or two or more heterologous self-assembling immunogenic peptides, or one or more oligomeric complexes, or a combination thereof.

[0151] The term “polypeptide nanoparticle” as used herein means a nanoparticle formed by self-assembly of polypeptides. The polypeptides comprising self-assembling immunogenic peptides may or may not include the target peptide inserted into them. In some embodiments, the polypeptide nanoparticle comprises two or more homologous polypeptides, or two or more heterologous polypeptides, or one or more oligomeric complexes, or a combination thereof.

[0152] The term “homologous self-assembling immunogenic peptide” as used herein means self-assembling immunogenic peptides in a multisubunit peptide that have identical self-assembling immunogenic peptides, and if present, identical target peptides. For example, if two self-assembling immunogenic peptides are identical, they are considered to be homologous self-assembling immunogenic peptides. If two self-assembling immunogenic peptides are identical, and the target peptides are also identical, they are also considered to be homologous self-assembling immunogenic peptides.

[0153] The term “heterologous self-assembling immunogenic peptide” as used herein means self-assembling immunogenic peptides in a multisubunit peptide that have different self-assembling immunogenic peptides, and if present, different or identical target peptides or when self-assembling immunogenic peptides are identical, but have different target peptides. For example, if two self- assembling immunogenic peptides are different, they are considered heterologous self-assembling immunogenic peptides. If two self-assembling immunogenic peptides are identical, but each has a different target peptide, they are considered heterologous self-assembling immunogenic peptides. If two self-assembling immunogenic peptides are different, and the target peptides, if present, are also different, they are considered heterologous self-assembling immunogenic peptides. The term “homologous polypeptide” as used herein means polypeptides that comprises identical self-assembling immunogenic peptides, and if present, identical target peptides. For example, if two polypeptides have identical self- assembling immunogenic peptides, they are considered to be homologous polypeptides. If two polypeptides have identical self-assembling immunogenic peptides and identical target peptides, they are also considered to be homologous polypeptides.

[0154] The term “heterologous polypeptide” as used herein means polypeptides that comprises different self-assembling immunogenic peptides, and if present, identical or different target peptides or when the polypeptides comprise identical self-assembling immunogenic peptides, but have different target peptides. For example, if two polypeptides have different or non-identical self-assembling immunogenic peptides, they are considered heterologous polypeptides. If two polypeptides have different or nonidentical self-assembling immunogenic peptides, but they either have identical or different target peptides, they are also considered heterologous polypeptides. If two polypeptides have identical self-assembling immunogenic peptides, but different target peptides, they are also considered to be heterologous polypeptides.

[0155] The term “upstream”, “amino-terminus”, or “N-terminus” has been used interchangeably in the context of amino acid sequences (protein, peptide, polypeptide, or any other sequence composed of amino acids) or the nucleic acid sequences (DNA, RNA or any other sequence composed of nucleotides) to mean amino end of an amino acid sequence or the 5-prime end of a nucleic acid sequence respectively.

[0156] The term “fragment” as used herein, whether in the context of a nucleic acid, nucleotide, protein, polypeptide, or peptide, means any length of the nucleic acid, protein, polypeptide, or peptide sequence except the full length of the respective nucleic acid, protein, polypeptide, or peptide sequence. Fragment includes such portions of nucleic acid, protein, polypeptide, or peptide that are capable of treating, preventing, diagnosing, improving symptoms of, and / or delay the onset of an infection, disease, disorder, and / or condition. A fragment is also understood to mean an immunogenic fragment of the protein, polypeptide or peptide, or a fragment of nucleic acid encoding an immunogenic fragment of the protein, peptide, or polypeptide.

[0157] The term “variant” as used herein, whether in the context of a nucleic acid, nucleotide, protein, polypeptide, or peptide sequence, means homologs, orthologs, paralogs, mutants or analogs of respective nucleic acid, protein, polypeptide, or peptide sequence.

[0158] The term “mutant” as used herein, whether in the context of a nucleic acid, nucleotide, protein, polypeptide, or peptide sequence, means a sequence which is not a wild type sequence. A mutant is also understood to mean a nucleic acid, nucleotide, protein, polypeptide, or peptide sequence that carries a mutation.

[0159] The term “mutation” as used herein means, a change or modification in the sequence of nucleic acid or amino acid in comparison to a reference sequence (for example, SEQ ID NO: 712 herein) and includes insertion (typically represented as “insKlOO” which means insertion of amino acid lysine at 100thposition), deletion (typically represented as “KlOOdel” or “KA100” which means deletion of amino acid lysine at 100thposition), substitution (typically represented as “K100V” which means substitution of amino acid lysine with amino acid valine), or a combination thereof. Mutations are introduced to impart desirable properties upon the nucleic acid, nucleotide, protein, polypeptide, or peptide sequence. This includes, for example, enabling the nucleic acid, nucleotide, protein, polypeptide, or peptide sequence to elicit an immune response while ensuring that any undesirable or deleterious effects are minimized or entirely removed.

[0160] The term “sequence” as used herein means nucleic acid sequences, nucleic acids, polynucleotides, amino acid sequences, proteins, polypeptides, or peptides, depending upon the context in which the term sequence is used, to mean a sequence of nucleotides or amino acids. In the context of nucleic acid, polynucleotide, or nucleotide sequence, the sequence is represented by a single letter code representing the nitrogenous base, for example A, T, G, C, or U. In the context of amino acid sequences, proteins, polypeptides, or peptides the sequence is represented by a single letter amino acid code as generally understood by persons skilled in the art. If the single letter amino acid code is represented by the letter “X”, it means the amino acid at that position is either absent or substituted by any other amino acid. In some embodiments, the sequences representing target sequence or target peptide, may contain a tag, for example, histidine tag, streptavidin tag etc, which may be deleted or removed from the respective sequence before employing the sequence in accordance with the present disclosure. In some embodiments, the sequences representing target sequence or target peptide may contain a signal sequence or signal peptide respectively, which may be deleted or removed from the respective sequence before employing the sequence in accordance with the present disclosure. The deleted or removed signal sequence or signal peptide may be employed in accordance with the present disclosure. The presence of tags, signal sequence or signal peptide, or other similar elements, may easily be recognized by those skilled in the art.

[0161] In the present disclosure, amino acids are typically represented by their single letter amino acid codes, for example A for alanine, R for arginine, N for asparagine, D for aspartic acid, C for cysteine, E for glutamic acid, Q for glutamine, G for glycine, H for histidine, I for isoleucine, L for leucine, K for lysine, M for methionine, F for phenylalanine, P for proline, S for serine, T for threonine, W for tryptophan, Y for tyrosine, V for valine and X for any of the amino acid. Alternatively, amino acids can also be represented by their three letter codes or by full name as generally known to a person skilled in the art.

[0162] The term “percentage identity”, “percent identity”, “% age identity”, or “% identity” have been used interchangeably and means the extent of identity between two sequences (e.g. nucleic acid sequences or amino acid sequences). Percent identity can be determined by aligning two sequences, introducing gaps to maximize identity between the sequences. Percent identity should generally be calculated between the same types of sequences for example nucleic acids, i.e. for DNA sequences or RNA sequences or amino acid sequences. The alignment of sequences (nucleic acid or amino acid) can be performed with the appropriate pair wise sequence alignment programs. Identity can be calculated between two sequences by multiplying the number of matches in the pair by 100 and dividing by the length of the aligned region, including gaps. Gaps at the end of sequences are not included, and internal gaps are included in the length. In some embodiments, the nucleic acid sequence or the amino acid sequence, as the case may be, shares at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity with the sequences disclosed herein. In some embodiments, the nucleic acid sequence or the amino acid sequence, as the case may be, shares at least 50% to 60%, at least 60% to 70%, at least 70% to 80%, at least 80% to 90%, or at least 90% to 100% identity with the sequences disclosed herein. The term “functional analog” or “functional analogue” have been used interchangeably, whether in the context of nucleic acid sequence or amino acid (protein or peptide) sequence, and mean all sequences which essentially performs similar function compared to the sequence being referred. For example, functional analogs of a target peptide include all sequences, irrespective of their percentage identity, which essentially perform at least one function similar to the function the target peptide performs.

[0163] The term “comparable equivalent” in the context of nucleic acid sequences, amino acid sequences, proteins, polypeptides, or peptides, as disclosed herein, means structurally or functionally similar or identical nucleic acid sequences, amino acid sequences, proteins, polypeptides, or peptides compared to the nucleic acid sequences or amino acid sequences being referred.

[0164] The term “capsid protein” or “nucleocapsid protein” as used herein means a protein that encapsidates or packages the viral genetic material or genome. The term is also understood to mean those proteins which are involved or participate in one or more of the following functions or activities, such as virus assembly, budding or release of virus, mediating attachment to and penetration into the host cells (especially in case of nonenveloped viruses), packaging the genome, etc. The capsid protein or nucleocapsid proteins are the proteins associated with viral capsid or viral nucleocapsid or viral core.

[0165] The term “B cell epitope” as used herein means a protein determinant that is recognized by B cell receptors (BCR) and capable of specific binding to an antibody or immunoglobulin.

[0166] The term “T cell epitope” as used herein means a protein determinant derived from an antigen which is presented by an antigen presenting cell (APC) through major histocompatibility complex (MHC) for recognition by a T cell receptor (TCR).

[0167] The term “papillomavirus” refers to a member of Papillomaviridae family, a family of non-enveloped DNA viruses. The term includes all papillomaviruses (PVs), such as human papillomavirus (HPV) and animal papillomavirus.

[0168] The term “human papillomavirus” or “HPV” refers to a DNA virus from the papillomaviridae family. The term includes, but is not limited to, HPV types HPV 1 , HPV2, HPV3, HPV4, HPV5, HPV6, HPV7, HPV8, HPV9, HPV10, HPV11, HPV12, HPV13, HPV14, HPV15, HPV16, HPV17, HPV18, HPV19, HPV20, HPV21, HPV22, HPV23, HPV24, HPV25, HPV26, HPV27, HPV28, HPV29, HPV30, HPV31, HPV32, HPV33, HPV34, HPV35, HPV36, HPV37, HPV38, HPV39, HPV40, HPV41, HPV42, HPV43, HPV44, HPV45, HPV46, HPV47, HPV48, HPV49, HPV50, HPV51, HPV52, HPV53, HPV54, HPV55, HPV56, HPV57, HPV58, HPV59, HPV60, HPV61, HPV62, HPV63, HPV64, HPV65, HPV66, HPV67, HPV68, HPV69, HPV70, HPV71, HPV72, HPV73, HPV74, HPV75, HPV76, HPV77, HPV78, HPV79, HPV80, HPV81, HPV82, HPV83, HPV84, HPV85, HPV86, HPV87, HPV88, HPV89, HPV90, HPV91, HPV92, HPV93, HPV94, HPV95, HPV96, HPV97, HPV98, HPV99, HPV100, HPV101, HPV102, HPV103, HPV104, HPV105, HPV106, HPV107, HPV108, HPV109, HPV11O, HPV111, HPV112, HPV113, HPV114, HPV115, HPV116, HPV117, HPV118, HPV119, HPV120, HPV121, HPV122, HPV123, HPV124, HPV125, HPV126, HPV127, HPV128, HPV129, HPV130, HPV131, HPV132, HPV133, HPV134, HPV135, HPV136, HPV137, HPV138, HPV139, HPV140, HPV141, HPV142, HPV143, HPV144, HPV145, HPV146, HPV147, HPV148, HPV148, HPV150, HPV151, HPV152, HPV153, HPV154, HPV155, HPV156, HPV157, HPV158, HPV159, HPV160, HPV161, HPV162, HPV163, HPV164, HPV165, HPV166, HPV167, HPV168, HPV169, HPV170, HPV171, HPV172, HPV173, HPV174, HPV175, HPV176, HPV177, HPV178, HPV179, HPV180, HPV181, HPV182, HPV183, HPV184, HPV185, HPV186, HPV187, HPV188, HPV189, HPV190, HPV191, HPV192, HPV193, HPV194, HPV195, HPV196, HPV197, HPV198, HPV199, HPV200, HPV201, HPV202, HPV203, HPV204, HPV205, HPV206, HPV207, HPV208, HPV209, HPV210, HPV211, HPV212, HPV213, HPV214, HPV215, HPV216, HPV217, HPV218, HPV219, HPV220, HPV221, HPV222, or hereafter discovered.

[0169] The term “LI protein”, “LI”, or “major capsid protein” have been used interchangeably to mean the major capsid protein of a papillomavirus which is expressed late into the viral infection cycle. It constitutes the major component of the papillomavirus viral capsid. The term LI protein also includes comparable equivalents of major capsid proteins of all papillomaviruses, including mutants, derivatives, variants, or functional analogs thereof.

[0170] The term “L2 protein”, “L2”, or “minor capsid protein” have been used interchangeably to mean the minor capsid protein which is one of the structural components of papillomavirus viral capsid and plays a role in the trafficking of the viral particle into the cells and escorting the viral genome towards the nucleus. The term L2 protein also includes comparable equivalents of minor capsid protein of all papillomaviruses, including mutants, derivatives, variants, or functional analogs thereof. The term “El protein” or “El” as used herein means one of the early proteins of papillomavirus which, among other functions, binds to the viral origin of DNA replication as hexameric complex facilitating oligomerization, and interacts with replication protein A. The term El protein also includes comparable equivalents of the El protein of all papillomaviruses, including mutants, derivatives, variants, or functional analogs thereof.

[0171] The term “E2 protein” or “E2” as used herein means one of the early proteins of papillomavirus which, among other functions, performs the role of a transcription and replication regulator. The term E2 protein also includes comparable equivalents of the E2 protein of all papillomaviruses, including mutants, derivatives, variants, or functional analogs thereof.

[0172] The term “E4 protein” or “E4” as used herein means one of the early proteins of papillomavirus which, among other functions, is involved in oligomerization, phosphorylation and proteolytic cleavage. It facilitates viral assembly and release. The term E4 protein also includes comparable equivalents of the E4 protein of all papillomaviruses, including mutants, derivatives, variants, or functional analogs thereof.

[0173] The term “E5 protein” or “E5” as used herein means one of the early proteins of papillomavirus which, among other functions, induces cell proliferation, activates growth factor receptors and inhibits apoptosis. The term E5 protein also includes comparable equivalents of the E5 protein of all papillomaviruses, including mutants, derivatives, variants, or functional analogs thereof.

[0174] The term “E6 protein” or “E6” as used herein means one of the early proteins of papillomavirus which, among other activities, mainly binds to the tumour suppressor protein p53 promoting ubiquitin-mediated degradation allowing cells to evade apoptosis. The term E6 protein also includes comparable equivalents of the E6 protein of all papillomaviruses, including mutants, derivatives, variants, or functional analogs thereof.

[0175] The term “E7 protein” or “E7” as used herein means one of the early proteins of papillomavirus which, among other activities, mainly binds tumour suppressor protein pRb (retinoblastoma protein) disrupting its function leading to uncontrolled growth of cells. The term E7 protein also includes comparable equivalents of the E7 protein of all papillomaviruses, including mutants, derivatives, variants, or functional analogs thereof.

[0176] Multisubunit nucleic acid sequence and multisubunit peptide

[0177] The multisubunit nucleic acid sequence encodes a multisubunit peptide. In the present disclosure, a multisubunit nucleic acid sequence includes two or more self-assembling immunogenic sequences, wherein each self-assembling immunogenic sequence is connected to an adjacent self- assembling immunogenic sequence by one or more cleavage sequence, wherein the multisubunit nucleic acid sequence further comprises one or more signal sequence upstream of one or more of the self-assembling immunogenic sequences.

[0178] In some embodiments, a multisubunit nucleic acid sequence includes two or more self-assembling immunogenic sequences each further comprising one or more target sequence, wherein each self-assembling immunogenic sequence is connected to an adjacent self-assembling immunogenic sequence by one or more cleavage sequence, wherein the multisubunit nucleic acid sequence further comprises one or more signal sequence upstream of one or more of the self-assembling immunogenic sequences.

[0179] In some embodiments, a multisubunit nucleic acid sequence includes two or more self-assembling immunogenic sequences, wherein the multisubunit nucleic acid sequences comprises either one or more self-assembling immunogenic sequences or one or more selfassembling immunogenic sequences each further comprising one or more target sequence, or a combination thereof, wherein each self-assembling immunogenic sequence is connected to an adjacent self-assembling immunogenic sequence by one or more cleavage sequence, and wherein the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the self-assembling immunogenic sequences.

[0180] In some embodiments, a multisubunit nucleic acid sequence includes two or more polynucleotide sequences wherein some or all polynucleotide sequences comprises either a self-assembling immunogenic sequence, or a self-assembling immunogenic sequence further comprising one or more target sequence, or a combination thereof, wherein each polynucleotide sequence is connected to an adjacent polynucleotide sequence by one or more cleavage sequence and wherein the multisubunit nucleic acid sequence further comprises one or more signal sequence upstream of one or more of the polynucleotide sequences.

[0181] In the present disclosure, a multisubunit peptide includes two or more selfassembling immunogenic peptides, wherein each self-assembling immunogenic peptide is connected to an adjacent self-assembling immunogenic peptide by one or more cleavage peptide, wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus (N-terminus) of one or more of the self-assembling immunogenic sequences.

[0182] In some embodiments, a multisubunit peptide includes two or more self-assembling immunogenic peptides each further comprising one or more target peptide, wherein each self-assembling immunogenic peptide is connected to an adjacent self-assembling immunogenic peptide by one or more cleavage peptide, wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus (N-terminus) of one or more of the self-assembling immunogenic sequences.

[0183] In some embodiments, a multisubunit peptide includes two or more self-assembling peptides, wherein the multisubunit peptide comprises either one or more self-assembling immunogenic peptides or one or more self-assembling immunogenic peptides each further comprising one or more target peptide, or a combination thereof, wherein each selfassembling immunogenic peptide is connected to an adjacent self-assembling immunogenic peptide by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus (N-terminus) of one or more of the self-assembling immunogenic peptides.

[0184] In some embodiments, a multisubunit peptide includes two or more polypeptides wherein some or all polypeptides comprises either a self-assembling immunogenic peptide, or a self-assembling immunogenic peptide further comprising one or more target peptide, or a combination thereof, wherein each polypeptide is connected to an adjacent polypeptide by one or more cleavage peptide and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus (N-terminus) of one or more of the polypeptides.

[0185] In some embodiments, the self-assembling immunogenic sequence is obtained or derived from LI protein of a papillomavirus. In some embodiments, the target sequence is obtained or derived from L2 protein, El protein, E2 protein, E4 protein, E5 protein, E6 protein, E7 protein, B cell epitope, T cell epitope, or a combination thereof of a papillomavirus.

[0186] In some embodiments, the signal sequence is present upstream of all or some selfassembling immunogenic sequences or polynucleotide sequences. In some embodiments, the signal sequence is present upstream of all self-assembling immunogenic sequences. In some embodiments, the signal sequence is present upstream of all polynucleotide sequences. In some embodiments, the signal sequence is present upstream of the first self- assembling immunogenic sequence or polynucleotide sequence. In some embodiments, the signal sequence is present upstream of the first self-assembling immunogenic sequence. In some embodiments, the signal sequence is present upstream of the first polynucleotide sequence. In some embodiments, the signal sequence is present upstream of some selfassembling immunogenic sequence or polynucleotide sequences. In some embodiments, the signal sequence is present upstream of some self-assembling immunogenic sequences. In some embodiments, the signal sequence is present upstream of some polynucleotide sequences. In some embodiments, the signal sequence is present upstream of each of the self-assembling immunogenic sequences or polynucleotide sequences. In some embodiments, the signal sequence is present upstream of each of the self-assembling immunogenic sequences. In some embodiments, the signal sequence is present upstream of each of the polynucleotide sequences.

[0187] In some embodiments, the multisubunit nucleic acid sequence either comprises a self-assembling immunogenic sequence or a self-assembling immunogenic sequence further comprising one or more target sequence, such that the total number of selfassembling immunogenic sequences in a multisubunit nucleic acid sequence are not more than 100. In some embodiments, the multisubunit nucleic acid sequences comprises two or more polynucleotide sequences such that the total number of polynucleotide sequences in a multisubunit nucleic acid sequence are not more than 100. Some exemplary illustrations of the multisubunit nucleic acid sequences are provided in figures 1, 3, 5, 6, & 7 and their representative encoded multisubunit peptides are provided in figures 2, 4, 8, 9, & 10 respectively.

[0188] The term “nucleic acid” as used herein means a polymer comprising two or more nucleotides for example, deoxyribonucleotides or ribonucleotides, either in an unmodified or modified form. The nucleic acid may be either single stranded or double stranded, linear, or circular. The term nucleic acid also encompasses fragments, variants, mutants, or codon optimized sequences of deoxyribonucleotides, ribonucleotides, or functional analogs thereof.

[0189] The term “nucleotide” as used herein means a ribonucleotide or deoxyribonucleotide. If the term nucleotide is used in the context of RNA, it refers to ribonucleotide, and if it is used in the context of DNA, it refers to deoxyribonucleotide. In some embodiments, the multisubunit nucleic acid sequence is a DNA, an RNA, or an mRNA. The multisubunit nucleic acid sequence may be few nucleotides long to several thousand nucleotides long.

[0190] Deoxyribonucleic acid (DNA)

[0191] The term “deoxyribonucleic acid” or “DNA” has been used interchangeably herein and means a polymer of deoxyribonucleotides. The DNA may be either single stranded or double stranded, linear, or circular.

[0192] In some embodiments, the multisubunit nucleic acid sequence is a DNA. In some embodiments, the DNA encodes a multisubunit peptide described herein.

[0193] Ribonucleic acid (RNA)

[0194] The term “ribonucleic acid” or “RNA” has been used interchangeably herein and means a polymer of ribonucleotides. The RNA may be either single stranded or double stranded, linear, or circular. The term RNA also includes messenger RNA (mRNA). In some embodiments, the multisubunit nucleic acid sequence is an mRNA.

[0195] In some embodiments, the mRNA encodes a multisubunit peptide as described herein.

[0196] In some embodiments, the mRNA is unmodified or modified or a combination of both. The modification may be in the nucleobase of the nucleotide, or sugar moiety of the nucleotide, or the phosphate of the nucleotide.

[0197] In some embodiments, mRNA is produced using recombinant expression system, or chemically synthesized or obtained through in vitro transcription. In some embodiments, the mRNA is obtained through one or more in vitro transcription (IVT) process or step. In some embodiments, the mRNA is obtained through a single in vitro transcription (IVT) process to step. In vitro transcription (IVT) is a laboratory process used to synthesize RNA molecules (for example, mRNA) from a DNA template enzymatically outside of a cell or in a cell free system. A single IVT process or step is understood to mean one complete cycle of an IVT reaction which produces mRNA molecules, each comprising at least two self- assembling immunogenic peptides or at least two polynucleotide sequences, as described herein, as against multiple IVT reactions that produces separate mRNA molecules, each comprising a single self-assembling immunogenic sequence or a single polynucleotide sequence. In some embodiments, the mRNA is circular. In other embodiments, the mRNA is linear.

[0198] In some embodiments, the mRNA is self-amplifying or self-replicating. Selfamplifying or self-replicating mRNA as used herein means an mRNA that self-replicate upon delivery into the cells. Such mRNAs typically contain a replicase sequence, usually derived from an alphavirus, which enables amplification of the original strand of mRNA encoding the protein of interest upon delivery into the cells (Beissert, Tim et al. Molecular Therapy (2020) 28:119-128).

[0199] The present disclosure provides mRNAs which are few hundred nucleotides long to several thousand nucleotides long. State of the art discourages using long mRNAs for vaccines and therapeutics. Longer mRNA molecules are more susceptible to degradation, which can compromise their stability and reduce their effectiveness in experimental and therapeutic context. Besides, longer mRNAs are also harder to transcribe accurately as the RNA polymerase used during the IVT reaction is inherently vulnerable to introduce errors within the transcribed mRNA. Additionally, long mRNA molecules are more prone to form complex secondary and tertiary structures, which can interfere with their intended function, reduce their efficiency, complicates the production process, and may even lead to unintended outcomes. Therefore, longer mRNAs are avoided in the art owing to their inherent complexities and challenges.

[0200] In some embodiments, mRNA is few hundred nucleotides long to several thousand nucleotides long. In some embodiments, mRNA is about 0.5 kb, 1 kb, 1.5 kb, 2 kb, 2.5 kb, 3 kb, 3.5 kb, 4 kb, 4.5 kb, 5 kb, 5.5 kb, 6 kb, 6.5 kb, 7.0 kb, 7.5 kb, 8 kb, 8.5 kb, 9 kb, 9.5 kb, 10 kb, 10.5 kb, 11 kb, 11.5 kb, 12 kb, 12.5 kb, 13 kb, 13.5 kb, 14 kb, 14.5 kb, 15 kb, 16 kb, 17 kb, 18 kb, 19 kb, 20 kb, 21 kb, 22 kb, 23 kb, 24 kb, 25 kb, 26 kb, 27 kb, 28 kb, 29 kb, 30 kb in length, or a fraction thereof. In some embodiments, mRNA is about 0.5 to 30 kb, 0.5 to 25 kb, 0.5 to 20 kb in length, or any range therein. In some embodiments, mRNA is about 1 to 20 kb, 1 to 18 kb, 1 to 16 kb, 1 to 14 kb, 1 to 12 kb, 1 to 10 kb, 1 to 9 kb, 1 to 8 kb, 1 to 7 kb, 1 to 6 kb, 1 to 5 kb in length, or any range therein.

[0201] In some embodiments, mRNA is about 0.5 kb to about 1 kb, about 1 kb to about 2 kb, about 2 kb to about 3 kb, about 3 kb to about 4 kb, about 4 kb to about 5 kb, about 5 kb to about 6 kb, about 6 kb to about 7 kb, about 7 kb to about 8 kb, about 8 kb to about 9 kb, about 9 kb to about 10 kb, about 10 kb to about 11 kb, about 11 kb to about 12 kb, about 12 kb to about 13 kb, about 13 kb to about 14 kb, about 14 kb to about 15 kb, about 15 kb to about 16 kb, about 16 kb to about 17 kb, about 17 kb to about 18 kb, about 18 kb to about 19 kb, about 19 kb to about 20 kb, about 20 kb to about 21 kb, about 21 kb to about 22 kb, about 22 kb to about 23 kb, about 23 kb to about 24 kb, about 24 kb to about 25 kb, about 25 kb to about 26 kb, about 26 kb to about 27 kb, about 27 kb to about 28 kb, about 28 kb to about 29 kb, about 29 kb to about 30 kb in length, or any range therein.

[0202] Self-assembling immunogenic sequence and self-assembling immunogenic peptide

[0203] The multisubunit nucleic acid sequence and multisubunit peptide includes selfassembling immunogenic sequence and self-assembling immunogenic peptide respectively. The self-assembling immunogenic sequence comprises a sequence of nucleotides, either deoxyribonucleotides or ribonucleotides, that encodes a self-assembling immunogenic peptide. The self-assembling immunogenic sequence includes codon optimized sequences, fragments, mutants, variants, comparable equivalents, functional analogs, or a combination thereof. In some embodiments, the self-assembling immunogenic sequence is a DNA or an RNA or an mRNA.

[0204] In some embodiments self-assembling immunogenic peptide may be a full-length protein or its fragment, mutants, variant, or functional analog thereof.

[0205] In some embodiments, the self-assembling immunogenic peptide is LI protein of a papillomavirus, including codon optimized nucleic acid sequences, fragments, mutants, variants, comparable equivalents, or functional analogs thereof.

[0206] Papillomavirus

[0207] Members of the genus papillomavirus consists of enveloped, double stranded DNA viruses. Human papillomaviruses (HPVs) are the most prominent papillomaviruses which are known to cause papillomas or warts, and are responsible for causing cervical cancer, vaginal cancer, vulvar cancer, penile cancer, anal cancer, and oropharyngeal cancer. Papillomaviruses include diverse group of viruses known to infect variety of hosts and are accordingly commonly referred as, for example, bovine papillomavirus (BPV), canine oral papillomavirus (COPV), cotton tail rabbit papillomavirus (CRPV), european elk papillomavirus (EEPV), rhesus monkey papillomavirus (RhPV), deer papillomavirus (DPV), and human papillomavirus (HPV).

[0208] The most extensively studied papillomaviruses are the human papillomaviruses (HPVs). The genome of HPV typically encodes six early proteins and two late proteins viz., El, E2, E4, E5, E6, E7, and LI and L2 respectively. The early proteins are responsible for viral replication and late proteins are part of viral capsid structure.

[0209] Papillomavirus genome encodes two structural proteins viz., LI protein and L2 protein which are expressed late into the viral infection cycle.

[0210] LI Protein (LI)

[0211] LI protein is a major capsid protein of papillomavirus. It is about 505 amino acids long with a molecular weight of approximately 56.3 kDa in HPV16 (variation in length and molecular weight may be possible among different genera, species, or types of papillomaviruses). LI protein forms pentamers, which self-assembles into a virus like particle which forms the major component of the HPV viral capsid.

[0212] In some embodiments, the self-assembling immunogenic peptide is obtained or derived from LI protein or comparable equivalents of a papillomavirus.

[0213] In some embodiments, the self-assembling immunogenic peptide is a full length LI protein or a fragment thereof of a papillomavirus or comparable equivalents, including mutants, derivatives, variants, or functional analogs thereof.

[0214] In some aspects, the multisubunit nucleic acid sequence and multisubunit peptide includes a self- assembling immunogenic sequence encoding LI protein of a papillomavirus and self-assembling immunogenic peptide representing LI protein of a papillomavirus, respectively. The self-assembling immunogenic sequence encoding LI protein of a papillomavirus comprises of a sequence of nucleotides, either deoxyribonucleotides or ribonucleotides, that encodes a self-assembling immunogenic peptide representing LI protein of a papillomavirus. In some embodiments, the selfassembling immunogenic sequence encoding LI protein of a papillomavirus is a DNA or an RNA or an mRNA.

[0215] Any LI protein of a papillomavirus that is capable of self-assembling into a nanoparticle (peptide nanoparticle or polypeptide nanoparticle) can be employed in accordance with the present disclosure.

[0216] In some embodiments, the LI protein is obtained or derived from a papillomavirus or comparable equivalents, including their codon optimized nucleic acid sequences, fragments, mutants, variants, or comparable equivalents thereof. In some embodiments, the LI protein comprises one or more mutations. In some embodiments, the mutation in the LI protein is a substitution replacing amino acid cysteine (C) with amino acid alanine (A), at the amino acid position 175 (i.e., C175A), compared to SEQ ID NO: 712. In some embodiments, the mutation in the LI protein is a substitution replacing amino acid cystine (C) with amino acid alanine (A), at the amino acid position 428 (i.e., C428A), compared to SEQ ID NO: 712. In some embodiments, the mutation in the LI protein is selected from C175A and C428A compared to SEQ ID NO: 712. In some embodiments, the LI protein comprises a substitution replacing the amino acid C with an amino acid A at a corresponding position in another LI protein of a different HPV type compared to the LI protein sequence identified as SEQ ID NO: 712. The amino acid position 175 in SEQ ID NO: 712 may not necessarily be the same position in another LI protein of a different HPV type when amino acid sequences of SEQ ID NO: 712 and the amino acid sequence of another LI protein of a different HPV type are aligned using appropriate pair wise sequence alignment programs well known to a skilled person.

[0217] In some embodiments, LI protein is obtained or derived from a human papillomavirus (HPV) selected from the group consisting of HPV1, HPV2, HPV3, HPV4, HPV5, HPV6, HPV7, HPV8, HPV9, HPV10, HPV11, HPV12, HPV13, HPV14, HPV15, HPV16, HPV17, HPV18, HPV19, HPV20, HPV21, HPV22, HPV23, HPV24, HPV25, HPV26, HPV27, HPV28, HPV29, HPV30, HPV31, HPV32, HPV33, HPV34, HPV35, HPV36, HPV37, HPV38, HPV39, HPV40, HPV41, HPV42, HPV43, HPV44, HPV45, HPV47, HPV48, HPV49, HPV50, HPV51, HPV52, HPV53, HPV54, HPV56, HPV57, HPV58, HPV59, HPV60, HPV61, HPV62, HPV63, HPV65, HPV66, HPV67, HPV68, HPV69, HPV70, HPV71, HPV72, HPV73, HPV74, HPV75, HPV76, HPV77, HPV78, HPV80, HPV81, HPV82, HPV83, HPV84, HPV85, HPV86, HPV87, HPV88, HPV89, HPV90, HPV91, HPV92, HPV93, HPV94, HPV95, HPV96, HPV97, HPV98, HPV99, HPV100, HPV101, HPV102, HPV103, HPV104, HPV105, HPV106, HPV107, HPV108, HPV109, HPV110, HPV111, HPV112, HPV113, HPV114, HPV115, HPV116, HPV117, HPV118, HPV119, HPV120, HPV121, HPV122, HPV123, HPV124, HPV125, HPV126, HPV127, HPV128, HPV129, HPV130, HPV131, HPV132, HPV133, HPV134, HPV135, HPV136, HPV137, HPV138, HPV139, HPV140, HPV141, HPV142, HPV143, HPV144, HPV145, HPV146, HPV147, HPV148, HPV148, HPV150, HPV151, HPV152, HPV153, HPV154, HPV155, HPV156, HPV157, HPV158, HPV159, HPV160, HPV161, HPV162, HPV163, HPV164, HPV165, HPV166, HPV167, HPV168, HPV169, HPV170, HPV171, HPV172, HPV173, HPV174, HPV175, HPV176, HPV177, HPV178, HPV179, HPV180, HPV181, HPV182, HPV183, HPV184, HPV185, HPV186, HPV187, HPV188, HPV189, HPV190, HPV191, HPV192, HPV193, HPV194, HPV195, HPV196, HPV197, HPV198, HPV199, HPV200, HPV201, HPV202, HPV203, HPV204, HPV205, HPV206, HPV207, HPV208, HPV209, HPV210, HPV211, HPV212, HPV213, HPV214, HPV215, HPV216, HPV217, HPV218, HPV219, HPV220, HPV221, HPV222, or a combination thereof.

[0218] In some embodiments, the LI protein is obtained or derived from a papillomavirus or comparable equivalents, including their codon optimized nucleic acid sequences, fragments, mutants, variants, or comparable equivalents thereof. In some embodiments, LI protein is obtained or derived from a human papillomavirus (HPV) selected from the group consisting of HPV5, HPV6, HPV8, HPV11, HPV16, HPV18, HPV26, HPV30, HPV31, HPV33, HPV34, HPV35, HPV39, HPV42, HPV43, HPV44, HPV45, HPV51, HPV52, HPV53, HPV56, HPV58, HPV59, HPV66, HPV67, HPV68, HPV69, HPV70, HPV73, HPV82, HPV85, HPV97, , or a combination thereof.

[0219] Exemplary LI protein includes, but not limited to, the one represented by the amino acid sequences having SEQ ID NOs: 76-1129 or 9409-9444, or comparable equivalents, including their codon optimized nucleic acid sequences, fragments, mutants, variants, or functional analogs thereof.

[0220] In some embodiments, the LI protein shares at least 50% identity with the sequences described herein or comparable equivalents, including their codon optimized nucleic acid sequences, fragments, mutants, variants, comparable equivalents, or functional analogs thereof.

[0221] Given the amino acid sequences of the LI protein, a person skilled in the art would be able to deduce all possible DNA or RNA sequences that encodes the above LI protein. Such DNA or RNA sequences are deemed to be incorporated in this disclosure. In some embodiments, the LI protein is encoded by a self-assembling immunogenic sequence which may be either a DNA, an RNA, or an mRNA.

[0222] Target Sequence and target peptide

[0223] The multisubunit nucleic acid sequence and the multisubunit peptide may include target sequence and target peptide respectively, wherein the target sequence and target peptide is obtained or derived from L2 protein, El protein, E2 protein, E4 protein, E5 protein, E6 protein, E7 protein, B cell epitope, T cell epitope, or a combination thereof of a papillomavirus. In some embodiments, target sequence and target peptide contains recurring target sequences and target peptides respectively. In some embodiments, the target sequence is a DNA or an RNA. In another embodiment, the target sequence is an mRNA. The target sequence may be modified or unmodified. The target sequence includes codon optimized sequences, fragments, mutants, variants, comparable equivalents, functional analogs, or combination thereof.

[0224] In some embodiments, the target sequence is a sequence of nucleotides that encodes a target peptide obtained or derived from L2 protein, El protein, E2 protein, E4 protein, E5 protein, E6 protein, E7 protein, B cell epitope, T cell epitope, or a combination thereof of a papillomavirus, including their codon optimized nucleic acid sequences, fragments, mutants, variants, comparable equivalents, or functional analogs thereof. In some embodiments, the target peptide is few amino acids long to several hundred amino acids long.

[0225] L2 Protein

[0226] L2 is the minor capsid protein of papillomavirus. It is about 473 amino acids long with a molecular weight of approximately 50.7 kDa in HPV16 (variation in length and molecular weight may be possible among different genera, species, or types of papillomaviruses). L2 is masked by LI and gets exposed after the LI protein undergoes conformational changes during the viral entry process. L2 is believed to aid in the stabilization of viral particle being part of the viral capsid structure, and plays a key role in the trafficking of the viral particle into the cells through a variety of interactions and escorting the viral genome towards the nucleus.

[0227] In some embodiments, the target peptide is obtained or derived from L2 protein or comparable equivalents of a papillomavirus.

[0228] In some embodiments, the L2 protein is a full length protein or a fragment thereof of a papillomavirus or comparable equivalents, including mutants, derivatives, variants, or functional analogs thereof.

[0229] Exemplary L2 protein includes, but not limited to, the one represented by the amino acid sequences having SEQ ID NOs: 1130-3675 or comparable equivalents, including their codon optimized nucleic acid sequences, fragments, mutants, variants, or functional analogs thereof. In some embodiments, the L2 protein shares at least 50% identity with the sequences described herein or comparable equivalents, including their codon optimized nucleic acid sequences, fragments, mutants, variants, functional analogs thereof.

[0230] Given the amino acid sequences of the L2 protein, a person skilled in the art would be able to deduce all possible DNA or RNA sequences that encodes the above L2 protein. Such DNA or RNA sequences are deemed to be incorporated in this disclosure. In some embodiments, the L2 protein is encoded by a self-assembling immunogenic sequence which may be either a DNA, an RNA, or an mRNA.

[0231] Papillomaviruses genome encodes several non-structural proteins during the early stages of infection cycle, and they are referred to as early (E) proteins. At least 6 early proteins are encoded by papillomavirus genome viz., El, E2, E4, E5, E6 & E7.

[0232] El Protein

[0233] El protein is one of the early proteins of papillomavirus. It is the largest and most conserved protein among the papillomaviruses. It is about 649 amino acids long with a molecular weight of approximately 72.9 kDa in HPV16 (variation in length and molecular weight may be possible among different genera, species, or types of papillomaviruses). El protein binds to the viral origin of DNA replication as hexameric complex which has helicase activity necessary for oligomerization. El protein also interacts with replication protein A.

[0234] In some embodiments, the target peptide is obtained or derived from El protein or comparable equivalents of a papillomavirus.

[0235] In some embodiments, the El protein is a full length protein or a fragment thereof of a papillomavirus or comparable equivalents, including mutants, derivatives, variants, or functional analogs thereof.

[0236] Exemplary El protein includes, but not limited to, the one represented by the amino acid sequences having SEQ ID NOs: 3676-4768 or comparable equivalents, including their codon optimized nucleic acid sequences, fragments, mutants, variants, or functional analogs thereof.

[0237] In some embodiments, the El protein shares at least 50% identity with the sequences described herein or comparable equivalents, including their codon optimized nucleic acid sequences, fragments, mutants, variants, functional analogs thereof. Given the amino acid sequences of the El protein, a person skilled in the art would be able to deduce all possible DNA or RNA sequences that encodes the above El protein. Such DNA or RNA sequences are deemed to be incorporated in this disclosure. In some embodiments, the El protein is encoded by a self-assembling immunogenic sequence which may be either a DNA, an RNA, or an mRNA.

[0238] E2 Protein

[0239] E2 protein of papillomavirus is a transcription and replication regulator. It is about 365 amino acids long with a molecular weight of approximately 41.8 kDa in HPV16 (variation in length and molecular weight may be possible among different genera, species, or types of papillomaviruses). E2 protein typically consists of an N-terminal transactivation domain of about 200 amino acids in length and a C-terminal DNA binding / dimerization domain of about 100 amino acids in length. These two domains are linked by a hinge of variable length in different papillomaviruses. E2 is a multifunctional protein. It activates and represses transcription in interaction with specific cellular factors / proteins. It plays an important role in viral DNA replication.

[0240] In some embodiments, the target peptide is obtained or derived from E2 protein or comparable equivalents of a papillomavirus.

[0241] In some embodiments, the E2 protein is a full length protein or a fragment thereof of a papillomavirus or comparable equivalents, including mutants, derivatives, variants, or functional analogs thereof.

[0242] Exemplary E2 protein includes, but not limited to, the one represented by the amino acid sequences having SEQ ID NOs: 4769-5850 or comparable equivalents, including their codon optimized nucleic acid sequences, fragments, mutants, variants, or functional analogs thereof.

[0243] In some embodiments, the E2 protein shares at least 50% identity with the sequences described herein or comparable equivalents, including their codon optimized nucleic acid sequences, fragments, mutants, variants, functional analogs thereof.

[0244] Given the amino acid sequences of the E2 protein, a person skilled in the art would be able to deduce all possible DNA or RNA sequences that encodes the above E2 protein. Such DNA or RNA sequences are deemed to be incorporated in this disclosure. In some embodiments, the E2 protein is encoded by a self-assembling immunogenic sequence which may be either a DNA, an RNA, or an mRNA. E4 Protein

[0245] E4 protein open reading frame lies within the larger E2 protein open reading frame. It is about 92 amino acids long with a molecular weight of approximately 10.1 kDa in HPV 16 (variation in length and molecular weight may be possible among different genera, species, or types of papillomaviruses). It is involved in oligomerization, phosphorylation and proteolytic cleavage. It facilitates viral assembly and release.

[0246] In some embodiments, the target peptide is obtained or derived from E4 protein or comparable equivalents of a papillomavirus.

[0247] In some embodiments, the E4 protein is a full length protein or a fragment thereof of a papillomavirus or comparable equivalents, including mutants, derivatives, variants, or functional analogs thereof.

[0248] Exemplary E4 protein includes, but not limited to, the one represented by the amino acid sequences having SEQ ID NOs: 5851-6415 or comparable equivalents, including their codon optimized nucleic acid sequences, fragments, mutants, variants, or functional analogs thereof.

[0249] In some embodiments, the E4 protein shares at least 50% identity with the sequences described herein or comparable equivalents, including their codon optimized nucleic acid sequences, fragments, mutants, variants, functional analogs thereof.

[0250] Given the amino acid sequences of the E4 protein, a person skilled in the art would be able to deduce all possible DNA or RNA sequences that encodes the above E4 protein. Such DNA or RNA sequences are deemed to be incorporated in this disclosure. In some embodiments, the E4 protein is encoded by a self-assembling immunogenic sequence which may be either a DNA, an RNA, or an mRNA.

[0251] E5 Protein

[0252] E5 protein is one of the early proteins of papillomavirus. It is about 83 amino acids long with a molecular weight of approximately 9.5 kDa in HPV 16 (variation in length and molecular weight may be possible among different genera, species, or types of papillomaviruses). It induces cell proliferation, activates growth factor receptors and inhibits apoptosis.

[0253] In some embodiments, the target peptide is obtained or derived from E5 protein or comparable equivalents of a papillomavirus. In some embodiments, the E5 protein is a full length protein or a fragment thereof of a papillomavirus or comparable equivalents, including mutants, derivatives, variants, or functional analogs thereof.

[0254] Exemplary E5 protein includes, but not limited to, the one represented by the amino acid sequences having SEQ ID NOs: 6416-6713 or comparable equivalents, including their codon optimized nucleic acid sequences, fragments, mutants, variants, or functional analogs thereof.

[0255] In some embodiments, the E5 protein shares at least 50% identity with the sequences described herein or comparable equivalents, including their codon optimized nucleic acid sequences, fragments, mutants, variants, functional analogs thereof.

[0256] Given the amino acid sequences of the E5 protein, a person skilled in the art would be able to deduce all possible DNA or RNA sequences that encodes the above E5 protein. Such DNA or RNA sequences are deemed to be incorporated in this disclosure. In some embodiments, the E5 protein is encoded by a self-assembling immunogenic sequence which may be either a DNA, an RNA, or an mRNA.

[0257] E6 Protein

[0258] E6 protein is one of the early proteins of papillomavirus. It is about 158 amino acids long with a molecular weight of approximately 19.2 kDa in HPV16 (variation in length and molecular weight may be possible among different genera, species, or types of papillomaviruses), depending upon the type of papillomavirus. It has the ability to bind zinc. E6 binds to the tumour suppressor protein p53 promoting ubiquitin-mediated degradation allowing cells to evade apoptosis. E6 also promotes telomerase activity leading to cell immortalization. E6 protein interacts with several cellular proteins leading to cell transformation and tumorigenesis.

[0259] In some embodiments, the target peptide is obtained or derived from E6 protein or comparable equivalents of a papillomavirus.

[0260] In some embodiments, the E6 protein is a full length protein or a fragment thereof of a papillomavirus or comparable equivalents, including mutants, derivatives, variants, or functional analogs thereof.

[0261] Exemplary E6 protein includes, but not limited to, the one represented by the amino acid sequences having SEQ ID NOs: 6714-7420 or comparable equivalents, including their codon optimized nucleic acid sequences, fragments, mutants, variants, or functional analogs thereof.

[0262] In some embodiments, the E6 protein shares at least 50% identity with the sequences described herein or comparable equivalents, including their codon optimized nucleic acid sequences, fragments, mutants, variants, functional analogs thereof.

[0263] Given the amino acid sequences of the E6 protein, a person skilled in the art would be able to deduce all possible DNA or RNA sequences that encodes the above E6 protein. Such DNA or RNA sequences are deemed to be incorporated in this disclosure. In some embodiments, the E6 protein is encoded by a self-assembling immunogenic sequence which may be either a DNA, an RNA, or an mRNA.

[0264] E7 Protein

[0265] E7 protein is one of the early proteins of papillomavirus. It is about 98 amino acids long with a molecular weight of approximately 11 kDa in HPV16 (variation in length and molecular weight may be possible among different genera, species, or types of papillomaviruses). E7 binds with the tumour suppressor protein pRb (retinoblastoma protein) disrupting its function leading to uncontrolled growth of cells. E7 protein is also known to interact with several cellular proteins, for example, cyclin dependent kinase inhibitors, TATA binding protein, histone deacetylase etc. affecting cellular transformation and promoting cell proliferation.

[0266] In some embodiments, the target peptide is obtained or derived from E7 protein or comparable equivalents of a papillomavirus.

[0267] In some embodiments, the E7 protein is a full length protein or a fragment thereof of a papillomavirus or comparable equivalent, including mutants, derivatives, variants, or functional analogs thereof.

[0268] Exemplary E7 protein includes, but not limited to, the one represented by the amino acid sequences having SEQ ID NOs: 7421-8044 or comparable equivalents, including their codon optimized nucleic acid sequences, fragments, mutants, variants, or functional analogs thereof.

[0269] In some embodiments, the E7 protein shares at least 50% identity with the sequences described herein or comparable equivalents, including their codon optimized nucleic acid sequences, fragments, mutants, variants, or functional analogs thereof. Given the amino acid sequences of the E7 protein, a person skilled in the art would be able to deduce all possible DNA or RNA sequences that encodes the above E7 protein. Such DNA or RNA sequences are deemed to be incorporated in this disclosure. In some embodiments, the E7 protein is encoded by a self-assembling immunogenic sequence which may be either a DNA, an RNA, or an mRNA.

[0270] B cell epitope

[0271] In some embodiments, the target peptide is obtained or derived from B cell epitope of a papillomavirus or comparable equivalents, including a fragment, mutant, derivative, variant, or functional analogs thereof.

[0272] Exemplary B cell epitopes include, but not limited to, the one represented by the amino acid sequences having SEQ ID NOs: 8045-8654 or comparable equivalents, including their codon optimized nucleic acid sequences, fragments, mutants, variants, or functional analogs thereof.

[0273] In some embodiments, the B cell epitopes share at least 50% identity with the sequences described herein or comparable equivalents, including their codon optimized nucleic acid sequences, fragments, mutants, variants, or functional analogs thereof.

[0274] Given the amino acid sequences of the B cell epitopes, a person skilled in the art would be able to deduce all possible DNA or RNA sequences that encodes the above B cell epitopes. Such DNA or RNA sequences are deemed to be incorporated in this disclosure. In some embodiments, the B cell epitopes are encoded by a target sequence which may be either a DNA, an RNA, or an mRNA.

[0275] T cell epitope

[0276] In some embodiments, the target peptide is obtained or derived from T cell epitope of a papillomavirus or comparable equivalents, including a fragment, mutant, derivative, variant, or functional analogs thereof.

[0277] Exemplary T cell epitopes include, but not limited to, the one represented by the amino acid sequences having SEQ ID NOs: 8655-9408 or comparable equivalents, including their codon optimized nucleic acid sequences, fragments, mutants, variants, or functional analogs thereof. In some embodiments, the T cell epitopes share at least 50% identity with the sequences described herein or comparable equivalents, including their codon optimized nucleic acid sequences, fragments, mutants, variants, or functional analogs thereof.

[0278] Given the amino acid sequences of the T cell epitopes, a person skilled in the art would be able to deduce all possible DNA or RNA sequences that encodes the above T cell epitopes. Such DNA or RNA sequences are deemed to be incorporated in this disclosure. In some embodiments, the T cell epitopes are encoded by a target sequence which may be either a DNA, an RNA, or an mRNA.

[0279] Linker sequence and linker peptide

[0280] The multisubunit nucleic acid sequence and multisubunit peptide includes linker sequence and linker peptide respectively. The linker sequence comprises a sequence of nucleotides, either deoxyribonucleotides or ribonucleotides, that encodes a linker peptide. The linker sequence includes codon optimized sequences, fragments, mutants, variants, comparable equivalents, functional analogs, or a combination thereof. In some embodiments, the linker sequence is a DNA, an RNA, or an mRNA.

[0281] In some embodiments, the linker peptide is present on the C-terminus, N-terminus, or both sides of the target peptide. In some embodiments, the linker peptide is on the N- terminus of the target protein. In some other embodiments, the linker peptide is on the C- terminus of the target peptide. In some embodiments, the linker peptide is on both N- terminus and C-terminus of the target peptide. In some embodiments, the linker peptide connects two cleavage peptides. In some embodiments, the linker peptide connects two signal peptides.

[0282] In some embodiments, the linker peptide is an amino acid linker, a foldon, a scaffold or a combination thereof. In some embodiments, the linker peptide is an amino acid linker. In some embodiments, the linker peptide is a foldon. In some embodiments, the linker peptide is a scaffold. In some embodiments, the linker peptide comprises a combination of an amino acid linker and a foldon. In some embodiments, the linker peptide comprises a combination of an amino acid linker and a scaffold. In some embodiments, the linker peptide comprises a combination of a foldon and a scaffold. In some embodiments, the linker peptide comprises a combination of an amino acid linker, a foldon, and a scaffold. In some embodiments, the amino acid linker comprises of about 2-49 amino acids, 2-40 amino acids, 2-30 amino acids, 2-20 amino acids, 2-15 amino acids, or 2-10 amino acids. In some embodiments, the amino acid linker comprises a glycine serine linker, a glycine proline linker, a glycine threonine linker, an alanine serine linker, any combination of two amino acid, or a combination thereof.

[0283] The glycine proline linker comprises of glycine (G) and proline (P) amino acids consecutively without any preference of order of appearance of either glycine or proline. In some embodiments, the glycine proline linker is 2-49 amino acids in length.

[0284] The glycine threonine linker comprises of glycine (G) and threonine (T) amino acids consecutively without any preference of order of appearance of either glycine or threonine. In some embodiments, the glycine threonine linker is 2-49 amino acids in length.

[0285] The alanine serine linker comprises of alanine (A) and serine (S) amino acids consecutively without any preference of order of appearance of either alanine or serine. In some embodiments, the alanine serine linker is 2-49 amino acids in length.

[0286] The glycine serine linker comprises of glycine (G) and serine (S) amino acids consecutively without any preference of order of appearance of either glycine or serine. In some embodiments, the glycine serine linker is 2-49 amino acids in length.

[0287] Exemplary amino acid linkers include, but not limited to, the ones represented by the following amino acid sequences or comparable equivalents, or their combinations, including their codon optimized nucleic acid sequences, fragments, mutants, variants, or functional analogs thereof:

[0288] GSG (SEQ ID NO: 1);

[0289] GSGG (SEQ ID NO: 2);

[0290] GGSGG (SEQ ID NO: 3);

[0291] GGSGGGGSGG (SEQ ID NO: 4);

[0292] GGSGGGGSGGGGSGG (SEQ ID NO: 5);

[0293] SGGSGG (SEQ ID NO: 6);

[0294] GGGGSGGGGS (SEQ ID NO: 7);

[0295] GGGGSGGGGSGGGGS (SEQ ID NO: 8).

[0296] In some embodiments, the amino acid linkers share at least 50% identity with the sequences described herein above or comparable equivalents, or a combination thereof, including their codon optimized nucleic acid sequences, fragments, mutants, variants, or functional analogs thereof.

[0297] Given the disclosed amino acid sequences of the amino acid linkers, a person skilled in the art would be able to deduce all possible DNA or RNA sequences that encodes the above amino acid linkers. Such DNA or RNA sequences are deemed to be incorporated in this disclosure. In some embodiments, the amino acid linker is encoded by an amino acid linker sequence which may be either a DNA, an RNA, or an mRNA.

[0298] In some embodiments, the linker peptide is a foldon. A foldon comprises of a sequence of amino acids encoded by a foldon sequence. The foldon sequence includes codon optimized sequences, fragments, mutants, variants, comparable equivalents, functional analogs, or a combination thereof. A foldon enables two or more homologous self-assembling immunogenic peptides or two or more homologous polypeptides to organise to form respective oligomeric complex. In some embodiments, foldon also helps in orientation of a self-assembling immunogenic peptides or polypeptide such that the domains or epitopes on the target peptide are exposed or displayed for interaction or communication with cells or biomolecules or immune system.

[0299] Exemplary foldons includes, but not limited to, the ones represented by the following amino acid sequences or comparable equivalents, or a combination thereof, including their codon optimized nucleic acid sequences, fragments, mutants, variants, or functional analogs thereof:

[0300] YIPEAPRDGQAYVRKDGEWVLLSTFL (SEQ ID NO: 9); HENEISHHAKEIERLQKEIERHKQSIKKLKQSE (SEQ ID NO: 10).

[0301] In some embodiments, the foldon shares at least 50% identity with the sequences described herein above or comparable equivalents, or a combination thereof, including their codon optimized nucleic acid sequences, fragments, mutants, variants, or functional analogs thereof.

[0302] Given the disclosed amino acid sequences of the foldons, a person skilled in the art would be able to deduce all possible DNA or RNA sequences that encodes the above foldons. Such DNA or RNA sequences are deemed to be incorporated in this disclosure. In some embodiments, the foldon is encoded by a foldon sequence which may be either a DNA, an RNA, or an mRNA. In some embodiments, the linker peptide is a scaffold. A scaffold comprises of a sequence of amino acids encoded by a scaffold sequence. The scaffold sequence includes codon optimized sequences, fragments, mutants, variants, comparable equivalents, functional analogs or a combination thereof. A scaffold provides structural and / or functional integrity or support to the target peptide and may also help in orientation of target peptide such that the domains or epitopes of the target peptide are exposed or displayed for interaction or communication with cells or biomolecules or immune system.

[0303] Exemplary scaffolds include, but not limited to, the ones represented by the following amino acid sequences or comparable equivalents, or a combination thereof, including their codon optimized nucleic acid sequences, fragments, mutants, variants, or functional analogs thereof: VDNKFNKEMRNAYWEIALLPNLNNQQKRAFIRSLYDDPSQSANLLAEAKKLNDA QAPK (SEQ ID NO: 11);

[0304] QDSTSDLIPAPPLSKVPLQQNFQDNQFHGKWYVVGKAGNHDLREDKDPRKMQAT IYELKEDKSYNVTNVRFVHKKCNYRIWTFVPGSQPGEFTLGNIKSWPGLTSWLVR VVSTNYNQHAMVFFKRVYQNRELFEITLYGRTKELTNELKENFIRFSKSLGLPENH IVFPVPIDQCIDGSAWSHPQFEK (SEQ ID NO: 12);

[0305] VSDVPRDLEVVAATPTSLLISWDAPAVTVRYYRITYGETGGNSPVQEFTVPGSKST ATISGLKPGVDYTITVYAVTGRGDSPASSKPISINYRT (SEQ ID NO: 13);

[0306] GCPRILMRCKQDSDCLAGCVCGPNGFCG (SEQ ID NO: 14);

[0307] MRGSHHHHHHGSDLGKKLLEAARAGQDDEVRILMANGADVNATDNDGYTPLHL AASNGHLEIVEVLLKNGADVNASDLTGITPLHLAAATGHLEIVEVLLKHGADVNA YDNDGHTPLHLAAKYGHLEIVEVLLKHGADVNAQDKFGKTAFDISIDNGNEDLA EILQ (SEQ ID NO: 15);

[0308] MRGSHHHHHHGSVKVKFFWNGEEKEVDTSKIVWVKRAGKSVLFIYDDNGKNGY GDVTEKDAPKELLDMLARAEREKKL (SEQ ID NO: 16);

[0309] MLPAPKNLVVSEVTEDSARLSWDDPAAFYESFLIQYQESEKVGEAIVLTVPGSERS YDLTGLKPGTEYTVSIYGVHNVYKDTNMRGLPLSAIFTTGGHHHHHH (SEQ ID NO: 17);

[0310] ETDICKLPKDEGTCRDFILKWYYDPNTKSCARFWYGGCGGNENKFGSQKECEKV CAPV (SEQ ID NO: 18); MIPGGLSEAKPATPEIQEIVDKVKPQLEEKTNETYGKLEAVQYKTQVVAGTNYYI KVRAGDNKYMHLKVFKSLPGQNEDLVLTGYQVDKNKDDELTGF (SEQ ID NO: 19);

[0311] PCSAFEFHCLSGECIHSSWRCDGGPDCKDKSDEENCA (SEQ ID NO: 20);

[0312] MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLS DYNIQKESTLHLVLRLRGG (SEQ ID NO: 21);

[0313] MGSIIFLEDRAFQGRIYGCTTDCPNLQPYFSRCNSIVVQSGCWMIYERPNYQGHQY FLRRGEYPDYQQWMGLSDSIRSCCLIPPHSGAYRMKIYDRDELRGQMSELTDDCL SVQDRFHLTEIHSLNVLEGSWILYEMPNYRGRQYLLRPGEYRRFLDWGAPNAKV GSLRRVMDLYLEHHHHHH (SEQ ID NO: 22);

[0314] AGHRIAWLLMMGHPRQQLAIIFGIGVSTLYRYFPA (SEQ ID NO: 23);

[0315] AFSKSEEARHSSLERECIEEICDHAEAWDIMM (SEQ ID NO: 24);

[0316] RECDYCGTDIEPGTGGMAVHGDGATTHFCSHRCAWDAMMGAEARNLEWTDTA R (SEQ ID NO: 25);

[0317] CSQNEYFDSLLHACIPCQLRCSGAPHRCAWDCMM (SEQ ID NO: 26);

[0318] EHIPGTLAARLSHRAAWDLMMHSLDASQGTATGPRGIFTAEDALKLVQLKQTGK

[0319] TFPTYKCGHRFAWDCMMGSGLNGAACFAVKIADLPVYSCECAIGFMGQRCEYKE (SEQ ID NO: 27);

[0320] ACYGHRCAWDCMMLGFSSGKCINSKCKCYK (SEQ ID NO: 28);

[0321] GEYVVEKVLDKRVVKGKVEYLLKWKGFSDEDNTWEPDENLDGHRLAWDFMM

[0322] ADVYEVEAILADRVNKNGINEYYIKWAGYDWYDNTWEPEQNLFGAGHRLAWW MMR (SEQ ID NO: 29);

[0323] AGTIKITQTRSAIGRLPAHKATLLGLGLRRIGHTVEREDGHRIAWDIMMVSFMVKV EG (SEQ ID NO: 30);

[0324] GIPCGESCGSPCISSAIGCSCKLINTNGSWHIVCYRN (SEQ ID NO: 31);

[0325] GKCPETFDAWYCLNDAHCFAVLINTNGSWHIVYSCECAIGFMGQRCEYKE (SEQ ID NO: 32);

[0326] QEEADRTVFVGNLEARVREEILYELFLQAGPLTKVTICKDREGKPKSFGFVCFKHP ESVSYAIALAGLINLNGSWIIVSGPSSG (SEQ ID NO: 33);

[0327] NEEDAGKMFVGGLSWDTSKKDLKDYFTKFGEVVDCTIKMDPNTGRSRGFGFILF KDAASVEKVLDAGLHNLNGSWIIPKKA (SEQ ID NO: 34);

[0328] SGNIFIKNLDKSIDNKALYDTFSAFGNILSCKVVCDEQGSKGYGFVHFETQEAAER AIAKMGLMNLNGSWVIVGRFKSRKE (SEQ ID NO: 35); PSRVVYLGSIPYDQTEEQILDLCSNVGPVINLKMMFDPQTGRSKGYAFIEFRDLESS ASAVGALGLYNLNGSWLICGYSSNSDISGVSLEHHHH (SEQ ID NO: 36);

[0329] LAILVFGYPETMANQVIAYFQEFGTILEDFEVLRKPQAMTVGLQDRQFVPIFSGNS WTKITYDNPASAVDALAEGLANFNGSWLLVIPYTKDAVERLQ (SEQ ID NO: 37); RLVNCNGSWLIGLDRPPYPGAKGEDIYNNVSRKAWDEWQKHQTMLINERRLNM MNAEDRKFLQQEMDKFLSGEDY (SEQ ID NO: 38);

[0330] FAVESIEKLRNRNGSWEILVKWRGWSPKYNTWEPEENIG (SEQ ID NO: 39); MRDFFVITNSLYNFNGSWYIKGAVLHVSPTQKRAFWVIADQENFIKQVNKNIEYV EKQASPAFLQRIVEIYQVKFEGKNVG (SEQ ID NO: 40).

[0331] In some embodiments, the scaffold shares at least 50% identity with the sequences described herein above or comparable equivalents, or a combination thereof, including their codon optimized nucleic acid sequences, fragments, mutants, variants, or functional analogs thereof.

[0332] Given the disclosed amino acid sequences of the scaffold, a person skilled in the art would be able to deduce all possible DNA or RNA sequences that encodes the above scaffold. Such DNA or RNA sequences are deemed to be incorporated in this disclosure. In some embodiments, the scaffold is encoded by the scaffold sequence which may be either a DNA, an RNA, or an mRNA.

[0333] In some embodiments, the linker peptide comprises an amino acid linker and a foldon. In some embodiments, the linker peptide comprises an amino acid linker followed by a foldon. In another embodiment, the linker peptide comprises a foldon followed by an amino acid linker. In some embodiments, the linker peptide comprises an amino acid followed by a foldon and another amino acid linker. In some embodiments, the linker peptide comprises a foldon followed by an amino acid linker and another foldon.

[0334] In some embodiments, the linker peptide comprises an amino acid linker and a scaffold. In some embodiments, the linker peptide comprises an amino acid linker followed by a scaffold. In some embodiments, the linker peptide comprises a scaffold followed by an amino acid linker. In some embodiments, the linker peptide comprises an amino acid linker followed by a scaffold and another amino acid linker. In some embodiments, the linker peptide comprises a scaffold followed by an amino acid linker and another scaffold. In some embodiments, the linker peptide comprises a foldon and a scaffold. In some embodiments, the linker peptide comprises a foldon followed by a scaffold. In some embodiments, the linker peptide comprises a scaffold followed by a foldon. In some embodiments, the linker peptide comprises a foldon followed by a scaffold and another foldon. In some embodiments, linker peptide comprises a scaffold followed by a foldon and another scaffold.

[0335] In some embodiments, the linker peptide comprises an amino acid linker, a foldon, and a scaffold. In some embodiments, the linker peptide consists of an amino acid linker followed by a foldon, and a scaffold. In some embodiments, the linker peptide comprises a foldon followed by an amino acid linker, and a scaffold. In some embodiments, the linker peptide comprises a scaffold followed by an amino acid linker, and a foldon. In some embodiments, the linker peptide comprises a first amino acid linker followed by a foldon, a second amino acid linker followed by scaffold, and a third amino acid linker. In some embodiments, the linker peptide comprises a first amino acid linker followed by a scaffold, a second amino acid linker followed by a foldon, and a third amino acid linker. In some embodiments, the linker peptide comprises a scaffold followed by a first amino acid linker, and a foldon followed by a second amino acid linker. In some embodiments, the linker peptide comprises a first amino acid linker followed by a scaffold, and a second amino acid linker followed by a foldon.

[0336] Cleavage sequence and cleavage peptide

[0337] The multisubunit nucleic acid sequence and multisubunit peptide includes cleavage sequence and cleavage peptide respectively. The cleavage sequence comprises of a sequence of nucleotides, either deoxyribonucleotides or ribonucleotides, that encodes a cleavage peptide. The cleavage sequence includes codon optimized sequences, fragments, mutants, variants, comparable equivalents, functional analogs, or a combination thereof. In some embodiments, the cleavage sequence is a DNA, an RNA, or an mRNA.

[0338] The cleavage peptide connects one self-assembling immunogenic peptide or polypeptide with another self-assembling immunogenic peptide or polypeptide respectively, for example, the adjacent self-assembling immunogenic peptide or polypeptide. In some embodiments, the cleavage peptide connects one self-assembling immunogenic peptide with another self-assembling immunogenic peptide, for example, adjacent self-immunogenic peptide. In some embodiments, the cleavage peptide connects one polypeptide with another polypeptide, for example, adjacent polypeptide. The cleavage peptide carries one or more cleavage sites. In some embodiments, the cleavage peptide comprises one or more cleavage peptides, for example cleavage peptide- 1, cleavage peptide-2 and so on. In some embodiments, the cleavage peptide optionally comprises a linker peptide between two cleavage peptides.

[0339] In some embodiments, the cleavage peptide facilitates the action of cellular proteases to cleave the multisubunit peptide into individual self-assembling immunogenic peptides or polypeptides or self cleaves into individual self-assembling immunogenic peptides or polypeptides. In some embodiments, the self-assembling immunogenic peptide or polypeptide, in addition to these peptides, also has some residues (amino acids) of cleavage peptide. Any cleavage peptide that is susceptible to the action of cellular proteases or a cleavage peptide that has the ability to undergo self cleavage can be employed in accordance with the present disclosure.

[0340] In some embodiments, the cleavage peptide is a substrate for cellular proteases. In some embodiments, the cleavage peptide is a substrate for golgi specific proteases. In some embodiments, the cleavage peptide is a self cleaving peptide. In some embodiments, the cleavage peptide comprises two or more cleavage peptides (for example, cleavage peptide- 1, cleavage peptide-2 and so on), optionally linked by a linker peptide, wherein one cleavage peptide is a substrate for cellular proteases and the other cleavage peptide is a self cleaving peptide. In some embodiments, the cleavage peptide is a golgi specific cleavage peptide i.e., susceptible to action of golgi specific proteases. In some embodiments, the cleavage peptide is a self cleaving peptide.

[0341] Exemplary cleavage peptide includes, but not limited to, the one represented by the following amino acid sequence or comparable equivalents, or a combination thereof, including their codon optimized nucleic acid sequences, fragments, mutants, variants, or functional analogs thereof:

[0342] RRKRSVS (SEQ ID NO: 41);

[0343] GIRRKRSVSH (SEQ ID NO: 42);

[0344] VQREKRAVGI (SEQ ID NO: 43);

[0345] SIRHKREPSV (SEQ ID NO: 44);

[0346] KRRQRRRPPQ (SEQ ID NO: 45);

[0347] KIRRRRDVVD (SEQ ID NO: 46);

[0348] HNRTKRSTDG (SEQ ID NO: 47); RKRRKRELET (SEQ ID NO: 48);

[0349] THRTRRSTSD (SEQ ID NO: 49);

[0350] SRRKRRSAST (SEQ ID NO: 50);

[0351] NLRRRRDLVD (SEQ ID NO: 51);

[0352] LRRRRRDAGN (SEQ ID NO: 52);

[0353] ATNFSLLKQAGDVEENPGP (SEQ ID NO: 53);

[0354] EGRGSLLTCGDVEENPGP (SEQ ID NO: 54);

[0355] QCTNYALLKLAGDVESNPGP (SEQ ID NO: 55).

[0356] In some embodiments, the cleavage peptide shares at least 50% identity with the sequence described herein above or comparable equivalents, or a combination thereof, including their codon optimized nucleic acid sequences, fragments, mutants, variants, or functional analogs thereof.

[0357] Given the above amino acid sequences of the cleavage peptide, a person skilled in the art would be able to deduce all possible DNA or RNA sequences that encodes the above cleavage peptide. Such DNA or RNA sequences are deemed to be incorporated in this disclosure. In some embodiments, the cleavage peptide is encoded by a cleavage sequence which may be either a DNA, an RNA, or an mRNA.

[0358] Signal sequence and signal peptide

[0359] The multisubunit nucleic acid sequence and multisubunit peptide includes a signal sequence and a signal peptide respectively. The signal sequence comprises of a sequence of nucleotides, either deoxyribonucleotides or ribonucleotides, that encodes a signal peptide. The signal sequence includes codon optimized sequences, fragments, mutants, variants, comparable equivalents, functional analogs, or a combination thereof. In some embodiments, the signal sequence is a DNA, an RNA, or an mRNA.

[0360] The signal peptide is present upstream (N-terminus or amino-terminus) of one or more self-assembling immunogenic peptides or polypeptides. In some embodiments, the signal peptide is present on the N-terminus of one or more self-assembling immunogenic peptide. In some embodiments, the signal peptide is present on the N-terminus of one or more polypeptide. In some embodiments, the signal peptide may be present on the N- terminus of all or some self-assembling immunogenic peptides or polypeptides. In some embodiments, the signal peptide is present on the N-terminus of all or some selfassembling immunogenic peptides. In some embodiments, the signal peptide is present on the N-terminus of all or some polypeptides. In some embodiments, the signal peptide is present on the N-terminus of the first self-assembling immunogenic peptide or the first polypeptide. In some embodiments, the signal peptide is present on the N-terminus of the first self-assembling immunogenic peptide. In some embodiments, the signal peptide is present on the N-terminus of the first polypeptide. In some embodiments, the signal peptide is present upstream (N-terminus) of some self-assembling immunogenic peptides or polypeptides. In some embodiments, the signal peptide is present on the N-terminus of some self-assembling immunogenic peptides. In some embodiments, the signal peptide is present on the N-terminus of some polypeptides. In some embodiments, the signal peptide is present upstream (N-terminus) of each of the self- assembling immunogenic peptides or polypeptides. In some embodiments, the signal peptide is present on the N-terminus of each of the self-assembling immunogenic peptides. In some embodiments, the signal peptide is present on the N-terminus of each of the polypeptides.

[0361] In some embodiments, the signal peptide transports the multisubunit peptide to cell organelles. In some embodiments, the signal peptide transports the multisubunit peptide to golgi body or golgi apparatus / complex. Any signal peptide that transports the multisubunit peptide to golgi bodies can be employed in accordance with the present disclosure. In some embodiments, the signal peptide is a golgi targeting signal peptide i.e., directs the multisubunit peptide to golgi complex.

[0362] Exemplary signal peptide includes, but not limited to, the one represented by the following amino acid sequence or comparable equivalents, or a combination thereof, including their codon optimized nucleic acid sequences, fragments, mutants, variants, or functional analogs thereof:

[0363] MPSSVSWGILLLAGLCCLVPVSLAEDPQGDAA (SEQ ID NO: 56); MGSSVSWGILLLAGLCCLVPVSLAEDPQGDAA (SEQ ID NO: 57); MASSVSWGILLLAGLCCLVPVSLAEDPQGDAA (SEQ ID NO: 58); MDMRAPAGIFGFLLVLFPGYRS (SEQ ID NO: 59);

[0364] MKWVTFISLLFLFSSAYS (SEQ ID NO: 60);

[0365] MDWTWRVFCLLAVTPGAHP (SEQ ID NO: 61);

[0366] MAWSPLFLTLITHCAGSWA (SEQ ID NO: 62); MTRLTVLALLAGLLASSRA (SEQ ID NO: 63); MARPLCTLLLLMATLAGALA (SEQ ID NO: 64); MRSLVFVLLIGAAFA (SEQ ID NO: 65); MSRLFVFILIALFLSAIIDVMS (SEQ ID NO: 66);

[0367] MGMRMMFIMFMLVVLATTVVS (SEQ ID NO: 67);

[0368] MRAFLFLTACISLPGVFG (SEQ ID NO: 68);

[0369] MKFQSTLLLAAAAGSALA (SEQ ID NO: 69);

[0370] MASSLYSFLLALSIVYIFVAPTHS (SEQ ID NO: 70);

[0371] MKTHYSSAILPILTLFVFLSINPSHG (SEQ ID NO: 71);

[0372] MESVSSLFNIFSTIMVNYKSLVLALLSVSNLKYARG (SEQ ID NO: 72);

[0373] MKAAQILTASIVSLLPIYTSA (SEQ ID NO: 73);

[0374] MIKLKFGVFFTVLLSSAYA (SEQ ID NO: 74);

[0375] MGVKVLFALICIAVAEA (SEQ ID NO: 75).

[0376] In some embodiments, the signal peptide shares at least 50% identity with the sequence described herein above or comparable equivalents, or a combination thereof, including their codon optimized nucleic acid sequences, fragments, mutants, variants, or functional analogs thereof.

[0377] Given the disclosed amino acid sequences of the signal peptide, a person skilled in the art would be able to deduce all possible DNA or RNA sequences that encodes the above signal peptide. Such DNA or RNA sequences are deemed to be incorporated in this disclosure. In some embodiments, the signal peptide is encoded by a signal sequence which may be either a DNA, an RNA, or an mRNA.

[0378] Synthesis of multisubunit nucleic acid sequences

[0379] Multisubunit nucleic acid sequence according to the present disclosure can be either a DNA or an RNA or an mRNA. The multisubunit nucleic acid sequence as described herein can be synthesized by molecular biology or genetic engineering techniques well known in the art, for example, using recombinant expression system, chemical synthesis, or in vitro transcription (IVT).

[0380] In some embodiments, the multisubunit nucleic acid sequence is obtained through a single or multiple IVT process or step. In some embodiments, the multisubunit nucleic acid sequence obtained through single or multiple IVT process or step is an mRNA. In some embodiments, the multisubunit nucleic acid sequence is obtained or synthesized through a single in vitro transcription (IVT) process or step.

[0381] In some embodiments, the multisubunit nucleic acid sequence is a messenger RNA (mRNA). The mRNA encodes a multisubunit peptide as described herein. Typically, an mRNA includes at least a coding region (which encodes the multisubunit peptide), a 5’ UTR, a 3’ UTR, a 5’ cap and a 3’ poly(A) tail. UTR (untranslated regions) flanks the coding region or open reading frame (ORF). The 5’ UTR and the 3’ UTR are sections of the mRNA before the start codon and after the stop codon respectively. The 5’ UTR has a cap (5’ cap) consisting of altered nucleotides. mRNA also contains a poly adenylated region at its 3’ end having adenine nucleotides called poly(A) tail.

[0382] In some embodiments, the mRNA is unmodified or modified or a combination of both. The modification may be in the nucleobase of the nucleotide, or sugar moiety of the nucleotide, or the phosphate of the nucleotide. In some embodiments, unmodified mRNA comprises naturally occurring nucleosides, for example, adenosine, guanosine, cytidine, and uridine. mRNA comprises one or more modified nucleosides, for example, adenosine analog, guanosine analog, cytidine analog, or uridine analog.

[0383] In some embodiments, the one or more modified nucleosides is a nucleoside analog selected from 2-aminoadenosine, 3-methyl adenosine, 7-deazaadenosine, 7- deazaguanosine, 8-oxoadenosine, or 8-oxoguanosine or a combination thereof.

[0384] In some embodiments, the one or more modified nucleosides is a uridine analog selected from propynyl-uridine, pseudouridine, C5 -bromouridine, C5-fluorouridine, C5- iodouridine, C5-propynyl-uridine, 5-aza-uridine, 2-thio-5-aza-uridine, 2-thio-uridine, 4- thio-pseudouridine, 2-thio-pseudouridine, 5-hydroxy-uridine, 3-methyl-uridine, 5- carboxymethyl-uridine, 1 -carboxymethyl-pseudouridine, l-methyl-3-(3-amino-3- carboxypropyl)pseudouridine, 2-thio-2 ’ -O-methyl-uridine, 5 -methoxycarbonylmethyl-2 ’ - O-methyl-uridine, 5-carboxymethylaminomethyl-2 ’ -O-methyl-uridine, 3 ,2 ’ -O-dimethyl- uridine, 5-propynyl-uridine, 1-propynyl-pseudouridine, 5-taurinomethyl-uridine, 1- taurinomethyl-pseudouridine, 5 -taurinomethyl-2-thio-uridine, 1 -taurino-4-thio- pseudouridine, 1-methyl-pseudouridine, 4-thio-l-methyl-pseudouridine, 2-thio-l-methyl- pseudouridine, 1 -methyl- 1 deaza-pseudouridine, 2-thio- 1 -methyl- 1 -deaza-pseudouridine, dihydro-uridine, dihydro-pseudouridine, 2-thio-dihydro-uridine, 2-thio-dihydro- pseudouridine, 2-methoxy-uridine, 2-methoxy-4-thio-uridine, 4-methoxy-pseudouridine, or 4-methoxy-2-thio-pseudouridine, or a combination thereof.

[0385] In some embodiments, the one or more modified nucleosides is a cytidine analog selected from 5-methylcytidine, C5-propynyl-cytidine, C5-methylcytidine, pseudoisocytidine, 1-methyl-pseudoisocytidine, pyrrolo-pseudoisocytidine, 4-thio- pseudoisocytidine, 4-thio- 1 -methyl-pseudoisocytidine, 4-thio- 1 -methyl- 1 -deaza- pseudoisocytidine, 1 -methyl- 1-1 deaza-pseudoisocytidine, 4-methoxy- 1 -methyl- pseudoisocytidine, or a combination thereof.

[0386] Methods for making modified nucleosides are well known in the art.

[0387] In some embodiments, the modified nucleoside is pseudouridine, for example, 1- methyl-pseudouridine, 1-propynyl-pseudouridine, 1-carboxymethyl-pseudouridine, 1- methyl-3-(3-amino-3-carboxypropyl) pseudouridine, 4-methoxy-pseudouridine, or 4- methoxy-2-thio-pseudouridine 4-thio-pseudouridine, 2-thio-pseudouridine, 4-thio- 1- methyl-pseudouridine, 2-thio-l-methyl-pseudouridine, dihydro-pseudouridine, or a combination thereof.

[0388] In some embodiments, mRNA is produced using recombinant expression system, chemically synthesized, or obtained through in vitro transcription.

[0389] In some embodiments, the multisubunit nucleic acid sequence is obtained or synthesized through a single or multiple IVT process or step. mRNAs according to the present disclosure may be synthesized via in vitro transcription (IVT). Briefly, IVT is typically performed with a DNA template containing a promoter, a pool of ribonucleotide triphosphates, a buffer system that may include DTT and magnesium ions, and an appropriate RNA polymerase (e.g., T3, T7, or SP6 RNA polymerase), DNase I, pyrophosphatase, and / or RNase inhibitor. The exact conditions may vary according to the specific application. Methods of making mRNA through IVT reaction is well known in the art (see for example, Beckert, Bertrand and Masquida, Benoit Methods in Molecular Biology (2011) 703, 29-41 ; Brunelle, Julie L. and Green Rachel Methods in Enzymology (2013) 530, 101-114; Kamakaka, Rohinton T. and Kraus W. Lee Current Protocols in Cell Biology (1999) 11.6.1-11.6.17; Kanwal, Fariha et al. Cellular Physiology and Biochemistry (2018) 48:1915-1927; WO2018157153;

[0390] WO202Q185811; W02022082001).

[0391] In some embodiments, the in vitro transcription occurs in a single batch. In some embodiments, IVT reaction includes capping and tailing reactions either co- transcriptionally or separately. A cap analog is added to the in vitro transcription reaction and will be incorporated at the 5’ end of the mRNA during the reaction. Alternative method of capping involves adding the cap post-transcriptionally through an enzymatic reaction. The poly (A) tail can be incorporated into the DNA template sequence, and thus the poly (A) tail will be incorporated into the mRNA by T7 RNA polymerase during the in vitro transcription. Alternative method of tailing involves adding the poly (A) tail post- transcriptionally through an enzymatic reaction. In some embodiments, capping and tailing reactions are performed co-transcriptionally i.e., during the IVT reaction. In some embodiments, capping and tailing reactions are performed separately from IVT reaction i.e., post transcriptionally. mRNA produced as a result of IVT reaction may be purified using techniques well known in the art, such as, centrifugation, filtration and / or chromatographic techniques. The purification of mRNA may be accomplished before capping and tailing steps are performed or after capping and tailing. The synthesized mRNA may be purified by ethanol precipitation or filtration or chromatography methods. In some embodiments, tangential flow filtration is used to purify mRNA. In some embodiments, mRNA is purified by chromatographic step. In other embodiments, mRNA is purified by a combination of filtration and chromatography steps.

[0392] In some embodiments, a suitable mRNA sequence is an mRNA sequence encoding a protein, peptide, polypeptide. In some embodiments, a suitable mRNA sequence is codon optimized for efficient expression in a host cell or organism. Codon optimization typically includes modifying a naturally-occurring or wild-type nucleic acid sequence encoding a peptide, polypeptide, or protein to achieve the highest possible expression of peptide, polypeptide, protein, or an antibody without altering the amino acid sequence.

[0393] In some embodiments, the mRNA is circular. In other embodiments, the mRNA is linear.

[0394] In some embodiments, the mRNA is self-amplifying or self-replicating.

[0395] In some embodiments, mRNA is few hundred nucleotides long to several thousand nucleotides long. In some embodiments, mRNA is about 0.5 kb, 1 kb, 1.5 kb, 2 kb, 2.5 kb, 3 kb, 3.5 kb, 4 kb, 4.5 kb, 5 kb, 5.5 kb, 6 kb, 6.5 kb, 7.0 kb, 7.5 kb, 8 kb, 8.5 kb, 9 kb, 9.5 kb, 10 kb, 10.5 kb, 11 kb, 11.5 kb, 12 kb, 12.5 kb, 13 kb, 13.5 kb, 14 kb, 14.5 kb, 15 kb, 16 kb, 17 kb, 18 kb, 19 kb, 20 kb, 21 kb, 22 kb, 23 kb, 24 kb, 25 kb, 26 kb, 27 kb, 28 kb, 29 kb, 30 kb in length, or a fraction thereof. In some embodiments, mRNA is about 0.5 to 30 kb, 0.5 to 25 kb, 0.5 to 20 kb in length or any range therein. In some embodiments, mRNA is about 1 to 20 kb, 1 to 18 kb, 1 to 16 kb, 1 to 14 kb, 1 to 12 kb, 1 to 10 kb, 1 to 9 kb, 1 to 8 kb, 1 to 7 kb, 1 to 6 kb, 1 to 5 kb in length, or any range therein.

[0396] In some embodiments, mRNA is about 0.5 kb to about 1 kb, about 1 kb to about 2 kb, about 2 kb to about 3 kb, about 3 kb to about 4 kb, about 4 kb to about 5 kb, about 5 kb to about 6 kb, about 6 kb to about 7 kb, about 7 kb to about 8 kb, about 8 kb to about 9 kb, about 9 kb to about 10 kb, about 10 kb to about 11 kb, about 11 kb to about 12 kb, about 12 kb to about 13 kb, about 13 kb to about 14 kb, about 14 kb to about 15 kb, about 15 kb to about 16 kb, about 16 kb to about 17 kb, about 17 kb to about 18 kb, about 18 kb to about 19 kb, about 19 kb to about 20 kb, about 20 kb to about 21 kb, about 21 kb to about 22 kb, about 22 kb to about 23 kb, about 23 kb to about 24 kb, about 24 kb to about 25 kb, about 25 kb to about 26 kb, about 26 kb to about 27 kb, about 27 kb to about 28 kb, about 28 kb to about 29 kb, about 29 kb to about 30 kb in length, or any range therein.

[0397] The multisubunit nucleic acid sequence as described herein, express multisubunit peptide.

[0398] Peptide nanoparticle or Polypeptide nanoparticle

[0399] The multisubunit peptide, encoded by the multisubunit nucleic acid, comprises multiple repeats of self-assembling immunogenic peptides or polypeptides interspersed with cleavage peptide (see illustration in figures). In some embodiments, the multisubunit peptide comprises multiple repeats of self-assembling immunogenic peptides. In some embodiments, the multisubunit peptide comprises multiple repeats of self-assembling immunogenic peptides further comprising one or more target peptide. In some embodiments, the multisubunit peptide comprises multiple repeats of self-assembling immunogenic peptides, self-assembling immunogenic peptides further comprising one or more target peptide, or a combination thereof each interspersed with a cleavage peptide. In some embodiments, the multisubunit peptide comprises multiple repeats of polypeptides interspersed with cleavage peptide, wherein the polypeptide comprises either a selfassembling immunogenic peptide or self-assembling immunogenic peptide further comprising one or more target peptide, or a combination thereof. In some embodiments, the total number of self-assembling immunogenic peptides or polypeptides present in a multisubunit peptide are up to 100 self-assembling immunogenic peptides or polypeptides, respectively. In some embodiments, the total number of self-assembling immunogenic peptides in a multisubunit peptide are up to 100 self-assembling immunogenic peptides. In some embodiments, the total number of polypeptides present in a multisubunit peptide are up to 100 polypeptides. In some embodiments, one or more self-assembling immunogenic peptides are homologous self-assembling immunogenic peptides. In some embodiments, one or more polypeptides are homologous polypeptides. In some embodiments, one or more self-assembling immunogenic peptides are heterologous self- assembling immunogenic peptides. In some embodiments, one or more polypeptides are heterologous polypeptides. In some embodiments, one or more self-assembling immunogenic peptides may be homologous self-assembling immunogenic peptides, heterologous self-assembling immunogenic peptides, or a combination thereof. In some embodiments, one or more polypeptides are homologous polypeptides, heterologous polypeptides, or a combination thereof.

[0400] A multisubunit peptide as described herein is encoded by the multisubunit nucleic acid sequence as described herein. Each multisubunit peptide comprises two or more selfassembling immunogenic peptides wherein the self- assembling immunogenic peptide is obtained or derived from LI protein of a papillomavirus. In some embodiments, the multisubunit peptide comprises two or more self-assembling immunogenic peptides each further comprising one or more target peptides, wherein the self- assembling immunogenic peptide is obtained or derived from LI protein of a papillomavirus, and wherein the target peptide is obtained or derived from L2 protein, El protein, E2 protein, E4 protein, E5 protein, E6 protein, E7 protein, B cell epitope, T cell epitope, or combination thereof of a papillomavirus.

[0401] In some embodiments, the multisubunit peptide comprises two or more polypeptides, wherein each polypeptide comprises either a self-assembling immunogenic peptide, a self-assembling immunogenic peptide each further comprising one or more target peptides, or a combination thereof, wherein the self-assembling immunogenic peptide is obtained or derived from LI protein of a papillomavirus, and wherein the target peptide is obtained or derived from L2 protein, El protein, E2 protein, E4 protein, E5 protein, E6 protein, E7 protein, B cell epitope, T cell epitope, or combination thereof of a papillomavirus.

[0402] The self-assembling immunogenic peptide or polypeptide is connected with another self-assembling immunogenic peptide or polypeptide respectively through a cleavage peptide. The multisubunit peptide includes a signal peptide upstream (N- terminus) of one or more self-assembling immunogenic peptides or polypeptides. In some embodiments, the multisubunit peptide includes a signal peptide upstream (N-terminus) of each of or some self-assembling immunogenic peptides or polypeptides. In some embodiments, the multisubunit peptide includes a signal peptide upstream (N-terminus) of one or more of self-assembling immunogenic peptide or polypeptide. In some embodiments, the multisubunit peptide includes a signal peptide upstream (N-terminus) of each self-assembling immunogenic peptide or polypeptide. In some embodiments, the multisubunit peptide includes a signal peptide upstream (N-terminus) of some selfassembling immunogenic peptides or polypeptides. In some embodiments, the multisubunit peptide includes a signal peptide upstream (N-terminus) of all selfassembling immunogenic peptides or polypeptides.

[0403] The signal peptide transports the multisubunit peptide to golgi body or golgi apparatus. The cellular proteases act on the cleavage sites present in the cleavage peptides or the cleavage peptide undergoes self cleavage and cleaves the multisubunit peptide into individual self-assembling immunogenic peptides (either self-assembling immunogenic peptides or self-assembling immunogenic peptides further comprising one or more target peptides, or both) or polypeptides comprising either a self-assembling immunogenic peptide or a self-assembling immunogenic peptide further comprising one or more target peptide or both, wherein the self- assembling immunogenic peptide is obtained or derived from LI protein of a papillomavirus, wherein the target peptide is obtained or derived from L2 protein, El protein, E2 protein, E4 protein, E5 protein, E6 protein, E7 protein, B cell epitope, T cell epitope, or combination thereof of a papillomavirus. The self-assembling immunogenic peptides or polypeptides may additionally also have some residues (amino acids) of cleavage peptide.

[0404] In some embodiments, the linker peptide is an amino acid linker, a foldon, a scaffold, or a combination thereof.

[0405] In some embodiments, the self-assembling immunogenic peptides are homologous self-assembling immunogenic peptides.

[0406] In some embodiments, the polypeptides are homologous polypeptides.

[0407] In some embodiments, two or more homologous self-assembling immunogenic peptides organise to form an oligomeric complex. In some embodiments, two or more polypeptides organise to form an oligomeric complex. In some embodiments, the oligomeric complex comprises at least two homologous self-assembling immunogenic peptides or at least two homologous polypeptides, at least three homologous selfassembling immunogenic peptides or at least three homologous polypeptides, at least four homologous self-assembling immunogenic peptides or at least four homologous polypeptides, at least five homologous self-assembling immunogenic peptides or at least five homologous polypeptides, or at least six homologous self- assembling immunogenic peptides or at least six homologous polypeptides and so on, respectively.

[0408] In some embodiments, the self-assembling immunogenic peptides are heterologous self-assembling immunogenic peptides. In some embodiments, the polypeptides are heterologous polypeptides.

[0409] A peptide nanoparticle is formed by self-assembly of two or more homologous self-assembling immunogenic peptides, or two or more heterologous self-assembling immunogenic peptides, or one or more oligomeric complexes formed by homologous selfassembling immunogenic peptides, or their combination.

[0410] In some embodiments, a peptide nanoparticle comprises homologous selfassembling immunogenic peptides, heterologous self-assembling immunogenic peptides, oligomeric complex formed by homologous self-assembling immunogenic peptide, or a combination thereof.

[0411] In some embodiments, the peptide nanoparticles are symmetrical, non-symmetrical, asymmetrical, or a combination thereof.

[0412] In some embodiments, the peptide nanoparticles are icosahedral, helical, spherical, rod-like, or a combination thereof.

[0413] In some embodiments, the peptide nanoparticles are enveloped or non-enveloped, or a combination thereof.

[0414] In some embodiments, the peptide nanoparticles are single layered or multilayered, or a combination thereof.

[0415] In some embodiments, the peptide nanoparticle comprises at least 2 or up to 500 self-assembling immunogenic peptides.

[0416] In some embodiments, the peptide nanoparticle comprises self- assembling immunogenic peptides between 2-5, 2-10, 2-20, 20-40, 40-60, 60-80, 80-100, 100-120, 120-140, 140-160, 160-180, 180-200, 200-220, 220-240, 240-260, 260-280, 280-300, 300- 320, 320-340, 340-360, 360-380, 380-400, 400-420, 420-440, 440-460, 460-480, or 480- 500.

[0417] In some embodiments, the peptide nanoparticle comprises self- assembling immunogenic peptides between 2-5, 2-10, 10-20, 20-30, 30-40, 40-50, 50-60, 60-70, 70- 80, 80-90, or 90-99.

[0418] In one of the embodiments, the peptide nanoparticle comprises at least 2 or up to 500 homologous self-assembling immunogenic peptides. In some embodiments, the peptide nanoparticle comprises at least 2 or up to 500 heterologous self-assembling immunogenic peptides.

[0419] In some embodiments, the peptide nanoparticle comprises two or more oligomeric complexes such that the total number of self- assembling immunogenic peptides in the peptide nanoparticle are not more than 500.

[0420] In some embodiments, the peptide nanoparticle comprises some homologous selfassembling immunogenic peptides and some heterologous self-assembling immunogenic peptides such that the total number of self-assembling immunogenic peptides in the peptide nanoparticle are not more than 500.

[0421] In some embodiments, the peptide nanoparticle comprises some homologous selfassembling immunogenic peptides and some oligomeric complexes such that the total number of self-assembling immunogenic peptides in the peptide nanoparticle are not more than 500.

[0422] In some embodiments, the peptide nanoparticle comprises some heterologous selfassembling immunogenic peptides and some oligomeric complexes such that the total number of self-assembling immunogenic peptides in the peptide nanoparticle are not more than 500.

[0423] In some embodiments, the peptide nanoparticle comprises some homologous selfassembling immunogenic peptides, some heterologous self- assembling immunogenic peptides, some oligomeric complexes, or their combination such that the total number of self-assembling immunogenic peptides in the peptide nanoparticle are not more than 500.

[0424] A polypeptide nanoparticle is formed by self-assembly of two or more homologous polypeptides, or two or more heterologous polypeptides, or one or more oligomeric complexes formed by homologous polypeptides, or their combination.

[0425] In some embodiments, a polypeptide nanoparticle comprises homologous polypeptides, heterologous polypeptides, oligomeric complex formed by homologous polypeptides, or a combination thereof.

[0426] In some embodiments, the polypeptide nanoparticles are symmetrical, non- symmetrical, asymmetrical, or a combination thereof.

[0427] In some embodiments, the polypeptide nanoparticles are icosahedral, helical, spherical, rod-like, or a combination thereof. In some embodiments, the polypeptide nanoparticles are enveloped or nonenveloped or a combination thereof.

[0428] In some embodiments, the polypeptide nanoparticles are single layered or multilayered or a combination thereof.

[0429] In some of the embodiments, the polypeptide nanoparticle comprises at least 2 or up to 500 polypeptides.

[0430] In some embodiments, the polypeptide nanoparticle comprises polypeptides between 2-5, 2-10, 2-20, 20-40, 40-60, 60-80, 80-100, 100-120, 120-140, 140-160, 160- 180, 180-200, 200-220, 220-240, 240-260, 260-280, 280-300, 300-320, 320-340, 340-360, 360-380, 380-400, 400-420, 420-440, 440-460, 460-480, or 480-500.

[0431] In some embodiments, the polypeptide nanoparticle comprises polypeptides between 2-5, 2-10, 10-20, 20-30, 30-40, 40-50, 50-60, 60-70, 70-80, 80-90, or 90-99.

[0432] In one of the embodiments, the polypeptide nanoparticle comprises at least 2 or up to 500 homologous polypeptides.

[0433] In some embodiments, the polypeptide nanoparticle comprises at least 2 or up to 500 heterologous polypeptides.

[0434] In some embodiments, the polypeptide nanoparticle comprises two or more oligomeric complexes such that the total number of polypeptides in the polypeptide nanoparticle are not more than 500.

[0435] In some embodiments, the polypeptide nanoparticle comprises some homologous polypeptides and some heterologous polypeptides such that the total number of polypeptides in the polypeptide nanoparticle are not more than 500.

[0436] In some embodiments, the polypeptide nanoparticle comprises some homologous polypeptides and some oligomeric complexes such that the total number of polypeptides in the polypeptide nanoparticle are not more than 500.

[0437] In some embodiments, the polypeptide nanoparticle comprises some heterologous polypeptides and some oligomeric complexes such that the total number of polypeptides in the polypeptide nanoparticle are not more than 500.

[0438] In some embodiments, the polypeptide nanoparticle comprises some homologous polypeptides, some heterologous polypeptides, some oligomeric complexes, or their combination such that the total number of polypeptides in the polypeptide nanoparticle are not more than 500. Lipid nanoparticles (LNP) composition

[0439] The multisubunit nucleic acid sequences as described herein may be encapsulated or formulated in a lipid nanoparticle composition.

[0440] In some embodiments, the lipid nanoparticle composition comprises lipid components, ionizable polymer, or a combination thereof and a multisubunit nucleic acid sequence as described herein.

[0441] In some embodiments, the lipid nanoparticle composition comprises lipid components such as a cationic lipid, a phospholipid, a sterol, a PEG-lipid and a multisubunit nucleic acid sequence as described herein.

[0442] In another embodiment, the lipid nanoparticle composition comprises an ionizable polymer, a cationic lipid, a phospholipid, a sterol, a PEG-lipid and a multisubunit nucleic acid sequence as described herein.

[0443] In some embodiments, a vaccine comprising the lipid nanoparticle composition is provided herein.

[0444] Lipid Components

[0445] Lipid components of the lipid nanoparticle compositions may include one or more lipids, such as a cationic lipid, a phospholipid, a sterol, and a PEG-lipid.

[0446] Cationic lipid

[0447] Cationic lipid refers to a lipid that has a net positive charge at a selected pH. Cationic lipids generally comprise a hydrophilic head group that carries the charge and a hydrophobic tail.

[0448] In some embodiments, the cationic lipid is a cationic lipid with an amine head group. The amine head group can be primary, secondary, tertiary, or quaternary. The cationic lipid may comprise one (monoamine) or more (polyamine) such amine groups.

[0449] In some embodiments, the cationic lipids are positively charged at pH below the pKa of the cationic lipid. In certain embodiments, the cationic lipids are neutral i.e., when pH is same or above the pKa of the cationic lipid. In some embodiments, the cationic lipids are positively charged at acidic pH i.e., pH 1.0 to pH 6.9. In certain embodiments, the cationic lipids are neutral at certain pH i.e., around physiological pH (pH 7.0 to pH 7.5). A cationic lipid that can exist in a positively charged or neutral form depending on the pH is commonly referred to as ionizable lipid. In some embodiments, the cationic lipids are ionizable such that they can exist in a positively charged or neutral form depending on the pH. In some embodiments, the cationic lipids are positively charged irrespective of the pH.

[0450] In some embodiments, the cationic lipid present in the lipid nanoparticle composition comprises a cationic lipid disclosed in US provisional applications viz., 63 / 575930; 63 / 575934; 63 / 575938; 63 / 575939; and 63 / 575942.

[0451] In some embodiments, the cationic lipid present in the lipid nanoparticle composition comprises a cationic lipid represented by formula (I) formula (I) or isomer, or salt thereof, wherein:

[0452] Ri and R2 are independently chosen from H, -OH, C1-10 alkyl, C2-10 alkenyl, C2-10 alkynyl, saturated or unsaturated C3-10 cycloalkyl, Ce-io aryl, saturated or unsaturated C3-10 heterocycloalkyl containing 1-4 heteroatoms, C5-10 heteroaryl containing 1-4 heteroatoms, -O-L7-R5, and -NReR?, or

[0453] Ri and R2 may combine together to form a saturated or unsaturated C3-10 cycloalkyl, Ce-io aryl, saturated or unsaturated C3-10 heterocycloalkyl containing 1-4 heteroatoms, or C5-10 heteroaryl containing 1-4 heteroatoms;

[0454] R3 and R4 are independently chosen from branched or unbranched C1-26 alkyl, C2-26 alkenyl, C2-26 alkynyl, -(CH2)m-A-(CH2)n-(CH3)y, and -CH((CH2)m-A-(CH2)n-(CH3)y)2, wherein each of m, n, and y is independently an integer ranging from 0 to 26;

[0455] Rs, Re, and R7 are independently chosen from H, -OH, C1-10 alkyl, C2-10 alkenyl, C2-10 alkynyl, saturated or unsaturated C3-10 cycloalkyl, Ce-io aryl, saturated or unsaturated C3-10 heterocycloalkyl containing 1-4 heteroatoms, and C5-10 heteroaryl containing 1-4 heteroatoms; each of Li, L2, L3, L4, L5, Le, and L7 is either absent or independently chosen from C1-10 alkylene, C2-10 alkenylene, and C2-10 alkynylene; Xi and X2 are independently chosen from -C(O)O-, -OC(O)-, -C(O)NH-, -NHC(O)-, - C(S)NH-, -NHC(S)-, -C(S)O-, -OC(S)-, -OC(O)NH-, -NHC(O)O-, -OP(O)(OH)O-, and - OP(O)(O-)O-;

[0456] A is H, a bond, saturated or unsaturated C3-10 cycloalkyl, Ce-io aryl, saturated or unsaturated C3-10 heterocycloalkyl containing 1-4 heteroatoms, or C5-10 heteroaryl containing 1-4 heteroatoms;

[0457] — Hc

[0458] Ai is \, -CH2-, -C(O)O-, -OC(O)-, -C(O)NH-, -NHC(O)-, -C(S)NH-, -NHC(S)-, - C(S)O-, -OC(S)-, -OC(O)NH-, -NHC(O)O-, -OP(O)(OH)O-, S, or N provided that when Ai is -CH2-, -C(O)O-, -OC(O)-, -C(O)NH-, -NHC(O)-, -C(S)NH-, -NHC(S)-, -C(S)O-, - OC(S)-, -OC(O)NH-, -NHC(O)O-, -OP(O)(OH)O-, or S, one of Ri or R2is absent; and wherein each alkyl, alkenyl, alkynyl, alkylene, alkenylene, alkynylene, cycloalkyl, aryl, heterocycloalkyl, or heteroaryl is independently optionally substituted with one or more substituent.

[0459] In some embodiments, the cationic lipid present in the lipid nanoparticle composition comprises a cationic lipid represented by formula (II) formula (II) or isomer, or salt thereof, wherein:

[0460] — represents either a single bond or a double bond;

[0461] Ri and R2 are independently chosen from H, -OH, C1-10 alkyl, C2-10 alkenyl, C2-10 alkynyl, saturated or unsaturated C3-10 cycloalkyl, Ce-io aryl, saturated or unsaturated C3-10 heterocycloalkyl containing 1-4 heteroatoms, C5-10 heteroaryl containing 1-4 heteroatoms, -O-L7-R5, and -NReR?, or

[0462] Ri and R2 may combine together to form a saturated or unsaturated C3-10 cycloalkyl, Ce-io aryl, saturated or unsaturated C3-10 heterocycloalkyl containing 1-4 heteroatoms, or C5-10 heteroaryl containing 1-4 heteroatoms;

[0463] R3 and R4 are independently chosen from, branched or unbranched, C1-26 alkyl, C2-26 alkenyl, C2-26 alkynyl, -(CH2)m-A-(CH2)n-(CH3)y, and -CH((CH2)m-A-(CH2)n-(CH3)y)2, wherein each of m, n, and y is independently an integer ranging from 0 to 26; Rs, Re, and R7 are independently chosen from H, -OH, C1-10 alkyl, C2-10 alkenyl, C2-10 alkynyl, saturated or unsaturated C3-10 cycloalkyl, Ce-io aryl, saturated or unsaturated C3-10 heterocycloalkyl containing 1-4 heteroatoms, and C5-10 heteroaryl containing 1-4 heteroatoms; each of Li, L2, L3, L4, Ls, Le, and L7 is either absent or independently chosen from C1-10 alkylene, C2-10 alkenylene, and C2-10 alkynylene;

[0464] Xi and X2 are independently chosen from -C(O)O-, -OC(O)-, -C(O)NH-, -NHC(O)-, - C(S)NH-, -NHC(S)-, -C(S)O-, -OC(S)-, -OC(O)NH-, -NHC(O)O-, -OP(O)(OH)O-, and - OP(O)(O-)O-;

[0465] A is H, a bond, saturated or unsaturated C3-10 cycloalkyl, Ce-io aryl, saturated or unsaturated C3-10 heterocycloalkyl containing 1-4 heteroatoms, or C5-10 heteroaryl containing 1-4 heteroatoms;

[0466] — Hc

[0467] Ai is \, -CH2-, -C(O)O-, -OC(O)-, -C(O)NH-, -NHC(O)-, -C(S)NH-, -NHC(S)-, - C(S)O-, -OC(S)-, -OC(O)NH-, -NHC(O)O-, -OP(O)(OH)O-, S, or N provided that when Ai is -CH2-, -C(O)O-, -OC(O)-, -C(O)NH-, -NHC(O)-, -C(S)NH-, -NHC(S)-, -C(S)O-, - OC(S)-, -OC(O)NH-, -NHC(O)O-, -OP(O)(OH)O-, or S, one of Ri or R2is absent; and wherein each alkyl, alkenyl, alkynyl, alkylene, alkenylene, alkynylene, cycloalkyl, aryl, heterocycloalkyl, or heteroaryl is independently optionally substituted with one or more substituent.

[0468] In some embodiments, the cationic lipid present in the lipid nanoparticle composition comprises a cationic lipid represented by formula (III) formula (III) or isomer, or salt thereof, wherein:

[0469] Ri and R2 are independently chosen from H, -OH, C1-10 alkyl, C2-10 alkenyl, C2-10 alkynyl, saturated or unsaturated C3-10 cycloalkyl, Ce-io aryl, saturated or unsaturated C3-10 heterocycloalkyl containing 1-4 heteroatoms, C5-10 heteroaryl containing 1-4 heteroatoms, -O-L7-R5, and -NReR?, or

[0470] Ri and R2 may combine together to form a saturated or unsaturated C3-10 cycloalkyl, Ce-io aryl, saturated or unsaturated C3-10 heterocycloalkyl containing 1-4 heteroatoms, or C5-10 heteroaryl containing 1-4 heteroatoms;

[0471] R3 and R4 are independently chosen from branched or unbranched C1-26 alkyl, C2-26 alkenyl, C2-26 alkynyl, -(CH2)m-A-(CH2)n-(CH3)y, and -CH((CH2)m-A-(CH2)n-(CH3)y)2, wherein each of m, n, and y is independently an integer ranging from 0 to 26;

[0472] Rs, Re, and R7 are independently chosen from H, -OH, C1-10 alkyl, C2-10 alkenyl, C2-10 alkynyl, saturated or unsaturated C3-10 cycloalkyl, Ce-io aryl, saturated or unsaturated C3-10 heterocycloalkyl containing 1-4 heteroatoms, and C5-10 heteroaryl containing 1-4 heteroatoms; each of Li, L2, L3, L4, L5 Le, and L7 is either absent or independently chosen from C1-10 alkylene, C2-10 alkenylene, and C2-10 alkynylene;

[0473] Xi and X2 are independently chosen from -C(O)O-, -OC(O)-, -C(O)NH-, -NHC(O)-, - C(S)NH-, -NHC(S)-, -C(S)O-, -OC(S)-, -OC(O)NH-, -NHC(O)O-, -OP(O)(OH)O-, and - OP(O)(O-)O-;

[0474] A is H, a bond, saturated or unsaturated C3-10 cycloalkyl, Ce-io aryl, saturated or unsaturated C3-10 heterocycloalkyl containing 1-4 heteroatom, or C5-10 heteroaryl containing 1-4 heteroatom;

[0475] — Hc

[0476] Ai is \, -CH2-, -C(O)O-, -OC(O)-, -C(O)NH-, -NHC(O)-, -C(S)NH-, -NHC(S)-, - C(S)O-, -OC(S)-, -OC(O)NH-, -NHC(O)O-, -OP(O)(OH)O-, S, or N provided that when Ai is -CH2-, -C(O)O-, -OC(O)-, -C(O)NH-, -NHC(O)-, -C(S)NH-, -NHC(S)-, -C(S)O-, - OC(S)-, -OC(O)NH-, -NHC(O)O-, -OP(O)(OH)O-, or S, one of Ri or R2is absent; and wherein each alkyl, alkenyl, alkynyl, alkylene, alkenylene, alkynylene, cycloalkyl, aryl, heterocycloalkyl, or heteroaryl is independently optionally substituted with one or more substituent.

[0477] In some embodiments, the cationic lipid present in the lipid nanoparticle composition comprises a cationic lipid represented by formula (IV) formula (IV) or isomer, or salt thereof, wherein:

[0478] = represents either a single bond or a double bond;

[0479] Ri and R2 are independently chosen from H, -OH, C1-10 alkyl, C2-10 alkenyl, C2-10 alkynyl, saturated or unsaturated C3-10 cycloalkyl, Ce-io aryl, saturated or unsaturated C3-10 heterocycloalkyl containing 1-4 heteroatoms, C5-10 heteroaryl containing 1-4 heteroatoms, -O-L7-R5, and -NReR?, or

[0480] Ri and R2 may combine together to form a saturated or unsaturated C3-10 cycloalkyl, Ce-io aryl, saturated or unsaturated C3-10 heterocycloalkyl containing 1-4 heteroatoms, or C5-10 heteroaryl containing 1-4 heteroatoms;

[0481] R3 and R4 are independently chosen from branched or unbranched C1-26 alkyl, C2-26 alkenyl, C2-26 alkynyl, -(CH2)m-A-(CH2)n-(CH3)y, and -CH((CH2)m-A-(CH2)n-(CH3)y)2, wherein each of m, n, and y is independently an integer ranging from 0 to 26;

[0482] Rs, Re, and R7 are independently chosen from H, -OH, C1-10 alkyl, C2-10 alkenyl, C2-10 alkynyl, saturated or unsaturated C3-10 cycloalkyl, Ce-io aryl, saturated or unsaturated C3-10 heterocycloalkyl containing 1-4 heteroatoms, and C5-10 heteroaryl containing 1-4 heteroatoms; each of Li, L2, L3, L4, L5, Le, and L7 is either absent or independently chosen from C1-10 alkylene, C2-10 alkenylene, and C2-10 alkynylene;

[0483] Xi and X2 are independently chosen from -C(O)O-, -OC(O)-, -C(O)NH-, -NHC(O)-, - C(S)NH-, -NHC(S)-, -C(S)O-, -OC(S)-, -OC(O)NH-, -NHC(O)O-, -OP(O)(OH)O-, and - OP(O)(O-)O-;

[0484] A is H, a bond, saturated or unsaturated C3-10 cycloalkyl, Ce-io aryl, saturated or unsaturated C3-10 heterocycloalkyl containing 1-4 heteroatom, or C5-10 heteroaryl containing 1-4 heteroatom;

[0485] -H

[0486] Ai is \, -CH2-, -C(O)O-, -OC(O)-, -C(O)NH-, -NHC(O)-, -C(S)NH-, -NHC(S)-, - C(S)O-, -OC(S)-, -OC(O)NH-, -NHC(O)O-, -OP(O)(OH)O-, S, or N provided that when Al is -CH2-, -C(0)0-, -0C(0)-, -C(O)NH-, -NHC(O)-, -C(S)NH-, -NHC(S)-, -C(S)O-, - OC(S)-, -OC(O)NH-, -NHC(O)O-, -OP(O)(OH)O-, or S, one of Ri or R2is absent; and wherein each alkyl, alkenyl, alkynyl, alkylene, alkenylene, alkynylene, cycloalkyl, aryl, heterocycloalkyl, or heteroaryl is independently optionally substituted with one or more substituent.

[0487] In some embodiments, the cationic lipid present in the lipid nanoparticle composition comprises a cationic lipid represented by formula (V) formula (V) or isomer, or salt thereof, wherein:

[0488] R3 and R4 are independently chosen from branched or unbranched C1-26 alkyl, C2.26 alkenyl, C2.26alkynyl, -(CH2)m-A-(CH2)n-(CH3)y, and -CH((CH2)m-A-(CH2)n-(CH3)y)2, wherein each of m, n, and y is independently an integer ranging from 0 to 26;

[0489] (Rio)qis chosen from H, -OH, optionally substituted C1-10 alkyl, C2-10 alkenyl, C2-10 alkynyl, saturated or unsaturated C3-10 cycloalkyl, Ce-io aryl, saturated or unsaturated C3-10 heterocycloalkyl containing 1-4 heteroatoms, C5-10 heteroaryl containing 1-4 heteroatoms, -O-L7-R5, and -NReR?, wherein q is an integer ranging from 0 to 5;

[0490] Rs, Re, and R7 are independently chosen from H, -OH, C1-10 alkyl, C2-10 alkenyl, C2-10 alkynyl, saturated or unsaturated C3-10 cycloalkyl, Ce-io aryl, saturated or unsaturated C3-10 heterocycloalkyl containing 1-4 heteroatoms, and C5-10 heteroaryl containing 1-4 heteroatoms; each of Li, L2, L3, L4, and L7 is either absent or independently chosen from C1-10 alkylene, C2-10 alkenylene, and C2-10 alkynylene;

[0491] Xi and X2are independently chosen from -C(O)O-, -OC(O)-, -C(O)NH-, -NHC(O)-, - C(S)NH-, -NHC(S)-, -C(S)O-, -OC(S)-, -OC(O)NH-, -NHC(O)O-, -OP(O)(OH)O-, and - OP(O)(O-)O-;

[0492] A is H, a bond, saturated or unsaturated C3-10 cycloalkyl, Ce-io aryl, saturated or unsaturated C3-10 heterocycloalkyl containing 1-4 heteroatoms, or C5-10 heteroaryl containing 1-4 heteroatoms; and wherein each alkyl, alkenyl, alkynyl, alkylene, alkenylene, alkynylene, cycloalkyl, aryl, heterocycloalkyl, heteroaryl is independently optionally substituted with one or more substituent.

[0493] In some embodiments, the cationic lipid present in the lipid nanoparticle composition comprises a cationic lipid represented by formula (VI) formula (VI) or isomer, or salt thereof, wherein:

[0494] R3 and R4 are independently chosen from branched or unbranched C1-26 alkyl, C2-26 alkenyl, C2-26 alkynyl, -(CH2)m-A-(CH2)n-(CH3)y, and -CH((CH2)m-A-(CH2)n-(CH3)y)2, wherein each of m, n, and y is independently an integer ranging from 0 to 26; each of Li, L2, L3, and L4 is either absent or independently chosen from C1-10 alkylene, C2- 10 alkenylene, and C2-10 alkynylene;

[0495] Xi and X2 are independently chosen from -C(O)O-, -OC(O)-, -C(O)NH-, -NHC(O)-, - C(S)NH-, -NHC(S)-, -C(S)O-, -OC(S)-, -OC(O)NH-, -NHC(O)O-, -OP(O)(OH)O-, and OP(O)(O-)O-;

[0496] A is H, a bond, saturated or unsaturated C3-10 cycloalkyl, Ce-io aryl, saturated or unsaturated C3-10 heterocycloalkyl containing 1-4 heteroatoms, or C5-10 heteroaryl containing 1-4 heteroatoms; and wherein each alkyl, alkenyl, alkynyl, alkylene, alkenylene, alkynylene, cycloalkyl, aryl, heterocycloalkyl, heteroaryl is independently optionally substituted with one or more substituent.

[0497] In some embodiments, the cationic lipid present in the lipid nanoparticle composition comprises a cationic lipid represented by formula (VII) or isomer, or salt thereof, wherein: = represents either a single bond or a double bond;

[0498] R3 and R4 are independently chosen from, branched or unbranched, C1-26 alkyl, C2-26 alkenyl, C2-26 alkynyl, -(CH2)m-A-(CH2)n-(CH3)y, and -CH((CH2)m-A-(CH2)n-(CH3)y)2, wherein each of m, n, and y is independently an integer ranging from 0 to 26;

[0499] (Rio)qis chosen from H, -OH, -CH2OH, C1-10 alkyl, C2-10 alkenyl, C2-10 alkynyl, saturated or unsaturated C3-10 cycloalkyl, Ce-io aryl, saturated or unsaturated C3-10 heterocycloalkyl containing 1-4 heteroatoms, C5-10 heteroaryl containing 1-4 heteroatoms, -O-Ls-Rs, and - NReR?, wherein q is an integer ranging from 0 to 5;

[0500] Rs, Re, and R7 are independently chosen from H, -OH, C1-10 alkyl, C2-10 alkenyl, C2-10 alkynyl, saturated or unsaturated C3-10 cycloalkyl, Ce-io aryl, saturated or unsaturated C3-10 heterocycloalkyl containing 1-4 heteroatoms, and C5-10 heteroaryl containing 1-4 heteroatoms; each of Li, L2, L3, L4, L5, L7, and Ls is either absent or independently chosen from C1-10 alkylene, C2-10 alkenylene, and C2-10 alkynylene;

[0501] Xi and X2 are independently chosen from -C(O)O-, -OC(O)-, -C(O)NH-, -NHC(O)-, - C(S)NH-, -NHC(S)-, -C(S)O-, -OC(S)-, -OC(O)NH-, -NHC(O)O-, -OP(O)(OH)O-, and - OP(O)(O-)O-;

[0502] A is H, a bond, saturated or unsaturated C3-10 cycloalkyl, Ce-io aryl, saturated or unsaturated C3-10 heterocycloalkyl containing 1-4 heteroatoms, or C5-10 heteroaryl containing 1-4 heteroatoms;

[0503] Z is -CH2-, O, N, or S;

[0504] Ai is -CH2-, -C(O)O-, -OC(O)-, -C(O)NH-, -NHC(O)-, -C(S)NH-, -NHC(S)-, -C(S)O-, - OC(S)-, -OC(O)NH-, -NHC(O)O-, -OP(O)(OH)O-, S, or N; and wherein each alkyl, alkenyl, alkynyl, alkylene, alkenylene, alkynylene, cycloalkyl, aryl, heterocycloalkyl, or heteroaryl is independently optionally substituted with one or more substituent.

[0505] In some embodiments, the cationic lipid present in the lipid nanoparticle composition comprises a cationic lipid represented by formula (VIII) formula (VIII) or isomer, or salt thereof, wherein:

[0506] R3 and R4 are independently chosen from branched or unbranched C1-26 alkyl, C2-26 alkenyl, C2-26 alkynyl, -(CH2)m-A-(CH2)n-(CH3)y, and -CH((CH2)m-A-(CH2)n-(CH3)y)2, wherein each of m, n, and y is independently an integer ranging from 0 to 26;

[0507] (Rio)qis chosen from H, -OH, -CH2OH, C1-10 alkyl, C2-10 alkenyl, C2-10 alkynyl, saturated or unsaturated C3-10 cycloalkyl, Ce-io aryl, saturated or unsaturated C3-10 heterocycloalkyl containing 1-4 heteroatoms, C5-10 heteroaryl containing 1-4 heteroatoms, -O-Ls-Rs, and - NReR?, wherein q is an integer ranging from 0 to 4;

[0508] Rs, Re, and R7 are independently chosen from H, -OH, C1-10 alkyl, C2-10 alkenyl, C2-10 alkynyl, saturated or unsaturated C3-10 cycloalkyl, Ce-io aryl, saturated or unsaturated C3-10 heterocycloalkyl containing 1-4 heteroatoms, and C5-10 heteroaryl containing 1-4 heteroatoms; each of Li, L2, L3, L4, L5, L7, and Ls is either absent or independently chosen from C1-10 alkylene, C2-10 alkenylene, or C2-10 alkynylene;

[0509] Xi and X2 are independently chosen from -C(O)O-, -OC(O)-, -C(O)NH-, -NHC(O)-, - C(S)NH-, -NHC(S)-, -C(S)O-, -OC(S)-, -OC(O)NH-, -NHC(O)O-, -OP(O)(OH)O-, and - OP(O)(O-)O-;

[0510] A is H, a bond, saturated or unsaturated C3-10 cycloalkyl, Ce-io aryl, saturated or unsaturated C3-10 heterocycloalkyl containing 1-4 heteroatoms, or C5-10 heteroaryl containing 1-4 heteroatoms;

[0511] Z is -CH2-, O, N, or S;

[0512] Ai is -CH2-, -C(O)O-, -OC(O)-, -C(O)NH-, -NHC(O)-, -C(S)NH-, -NHC(S)-, -C(S)O-, - OC(S)-, -OC(O)NH-, -NHC(O)O-, -OP(O)(OH)O-, S, or N; and wherein each alkyl, alkenyl, alkynyl, alkylene, alkenylene, alkynylene, cycloalkyl, aryl, heterocycloalkyl, or heteroaryl is independently optionally substituted with one or more substituent.

[0513] In some embodiments, the cationic lipid represented by formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), or formula (VIII) are substituted with substituents independently chosen from H, OH, Cl, Br, I, O, S, N, P, optionally substituted C1-6 alkoxy, optionally substituted C1-6 alkyl, optionally substituted C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted saturated or unsaturated C3-10 cycloalkyl, optionally substituted Ce-io aryl, optionally substituted saturated or unsaturated C3-10 heterocycloalkyl containing 1-4 heteroatoms, optionally substituted C5-10 heteroaryl containing 1-4 heteroatoms or combination thereof. In some embodiments, substituent may further be substituted with H, -OH, Cl, Br, I, or Ci- 6 hydroxyalkyl.

[0514] In some embodiments, N:P ratio or cationic lipid to nucleic acid ratio in LNP formulation is between 1 to 18, between 1 to 17, between 1 to 16, between 1 to 15, between 1 to 14, between 1 to 13, between 1 to 12, between 1 to 11, between 1 to 10, between 1 to 9, between 1 to 8, between 1 to 7, between 1 to 6, between 1 to 5, between 1 to 4, between 1 to 3, between 1 to 2, or any range therein.

[0515] In some embodiments, N:P ratio or cationic lipid to nucleic acid ratio in LNP formulation is about 18, about 17, about 16, about 15, about 14, about 13, about 12, about 11, about 10, about 9, about 8, about 7, about 6, about 5, about 4, about 3, about 2, about 1, or any portion or fraction thereof.

[0516] In some embodiments, other exemplary cationic lipid for use in the lipid nanoparticle compositions include, but are not limited to, N,N-dioleyl-N,N- dimethylammonium chloride (DODAC); N-(2,3-dioleyloxy)propyl)-N,N,N- trimethylammonium chloride (DOTMA); N,N-distearyl-N,N-dimethylammonium bromide(DDAB); N-(2,3dioleoyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTAP); 3-(N — (N’,N’-dimethylaminoethane)-carbamoyl)cholesterol (DC-Chol), N-(l- (2,3-dioleoyloxy)propyl)N-2-(sperminecarboxamido)ethyl)-N,N- dimethylammoniumtrifluoracetate (DOSPA), dioctadecylamidoglycyl carboxyspermine (DOGS), 1,2-dioleoyl-

[0517] 3 -dimethylammonium propane (DODAP), N,N-dimethyl-2,3-dioleoyloxy)propylamine (DODMA), N-(l ,2-dimyristyloxyprop-3-yl)-N,N-dimethyl-N-hydroxyethyl ammonium bromide (DMRIE), l,2-dilinoleyloxy-N,N-dimethylaminopropane (Dlin-DMA), 3- dimethylamino-2-(cholest-5-en-3-beta-oxybutan-4-oxy)-l-(cis,cis-9,12-oc- tadecadienoxy)propane (Clin-DMA), 2-[5’-(cholest-5-en-3-beta-oxy)-3’-oxapentoxy)-3- dimethyl-l-(cis,cis-9’,12’-octadecadienoxy)propane (CpLin-DMA), 2,3-Dilinoleoyloxy- N,N-dimethylpropylamine (Dlin-DAP), l,2-N,N’-Dilinoleylcarbamyl-3- dimethylaminopropane (Dlincarb-DAP), l,2-Dilinoleoylcarbamyl-3- dimethylaminopropane (Dlin-CD AP) , 2 ,2-dilinoleyl-4-dimethylaminomethyl- [1,3]- dioxolane (Dlin-K-DMA), heptatriaconta-6,9,28,31-tetraen-19-yl 4- (dimethylamino)butanoate (Dlin-MC3-DMA), heptadecane-9-yl 8-[2-hydroxyethyl-(6- oxo-6-undecoxyhexyl)amino]octanoate (SM-102), 6-[6-(2-hexyldecanoyloxy)hexyl-(4- hydroxybutyl)amino]hexyl 2-hexyldecanoate (ALC-0315), nonyl 8-[(8-heptadecan-9- yloxy-8-oxooctyl)-(2-hydroxyethyl)amino]octanoate (SLP-0001), or a combination thereof.

[0518] Methods of making cationic lipid and / or ionizable lipid or imparting the cationic lipid the ability to behave as an ionizable lipid are well known in the art (WO2005121348; W02009127060; W02009086558; W02010042877; W02010144740; WO2011075656; WO2017049245; WO2017075531; WO2018118102; WO2015199952; Reynier P. et al. Journal of Drug Targeting (2004) 12: 25-38; Sabnis, Staci et al. Molecular Therapy (2018) 26: 1509-1519). In some embodiments, the cationic lipid present in the lipid nanoparticle composition comprises a cationic lipid disclosed in published patent application viz., WO2019 / 152557; WO2019 / 232095; WO2021 / 077067; WO2019 / 089828; US2019 / 0240354; US2010 / 0130588; US2021 / 0087135; US2021 / 0128488; US2020 / 0121809; US2013 / 0108685; US2013 / 0195920; US2015 / 0005363; US2014 / 0308304; US 2017 / 0210697; and US2013 / 0053572.

[0519] The proportion of cationic lipid present in the lipid nanoparticle compositions is from about 10 mol % to about 70 mol % or any range therein.

[0520] In some embodiments, the proportion of cationic lipid present in the lipid nanoparticle compositions is from about 10 mol % to about 70 mol %, from about 10 mol % to about 65 mol %, from about 10 mol % to about 60 mol %, from about 10 mol % to about 55 mol %, from about 10 mol % to about 50 mol %, or any range therein.

[0521] In some embodiments, the proportion of cationic lipid present in the lipid nanoparticle compositions is about 10 mol %, about 11 mol %, about 12 mol %, about 13 mol %, about 14 mol %, about 15 mol %, about 16 mol %, about 17 mol %, about 18 mol %, about 19 mol %, about 20 mol %, about 21 mol %, about 22 mol %, about 23 mol %, about 24 mol %, about 25 mol %, about 26 mol %, about 27 mol %, about 28 mol %, about 29 mol %, about 30 mol %, about 31 mol %, about 32 mol %, about 33 mol %, about 34 mol %, about 35 mol %, about 36 mol %, about 37 mol %, about 38 mol %, about 39 mol %, about 40 mol %, about 41 mol %, about 42 mol %, about 43 mol %, about 44 mol %, about 45 mol %, about 46 mol %, about 47 mol %, about 48 mol %, about 49 mol %, about 50 mol % about 51 mol %, about 52 mol %, about 53 mol %, about 54 mol %, about 55 mol %, about 56 mol %, about 57 mol %, about 58 mol %, about 59 mol %, about 60 mol %, about 61 mol %, about 62 mol %, about 63 mol %, about 64 mol %, about 65 mol %, about 66 mol %, about 67 mol %, about 68 mol %, about 69 mol %, about 70 mol % or any portion or fraction thereof. In some embodiments, the proportion of cationic lipid present in the lipid nanoparticle compositions is about 10 mol% to about 20 mol%, about 20 mol% to about 30 mol%, about 30 mol% to about 40 mol%, about 40 mol% to about 50 mol%, about 50 mol% to about 60 mol%, or about 60 mol% to about 70 mol%.

[0522] Phospholipids

[0523] Phospholipid includes a lipid containing a hydrophilic head with a phosphate group and a hydrophobic tail composed of fatty acid chains attached to a glycerol or sphingosine backbone.

[0524] Exemplary phospholipids for use in the lipid nanoparticle compositions include, but are not limited to, l,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2- dimyristoyl-sn-glycero-phosphocholine (DMPC), 1 ,2-dioleoyl-sn-glycero-3- phosphocholine (DOPC), l,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2- distearoyl-sn-glycero-3-phosphocholine (DSPC), 1 ,2-diundecanoyl-sn-glycero- phosphocholine (DUPC), l-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2- di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 Diether PC), l-oleoyl-2- cholesterylhemisuccinoyl-sn-glycero-3-phosphocholine (OchemsPC), 1 -hexadecyl-sn- glycero-3-phosphocholine (C16 Lyso PC), l,2-dilinolenoyl-sn-glycero-3-phosphocholine, 1 ,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1 ,2-didocosahexaenoyl-sn-glycero-3- phosphocholine, l,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2- diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16.0 PE), 1,2-distearoyl-sn-glycero- 3-phosphoethanolamine, l,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2- dilinolenoyl-sn-glycero-3-phosphoethanolamine, l,2-diarachidonoyl-sn-glycero-3- phosphoethanolamine, 1 ,2-didocosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1 ,2- dioleoyl-sn-glycero-3-phospho-rac-(l -glycerol) sodium salt (DOPG), l-myristoyl-2- stearoyl-sn-glycero-3-phosphocholine (MSPC), l-palmitoyl-2-myristoyl-sn-glycero-3- phosphocholine (PMPC), l-palmitoyl-2-stearoyl-sn-glycero-3-phosphocholine (PSPC), 1- stearoyl-2-myristoyl-sn-glycero-3-Phosphocholine (SMPC), l-Stearoyl-2-palmitoyl-sn- glycero-3-phosphocholine (SPPC), l-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine (SOPC), l-stearoyl-2-docosahexaenoyl-sn-glycero-3-phosphocholine (SDPC), sphingomyelin, or a combination thereof.

[0525] The proportion of phospholipid present in the lipid nanoparticle compositions is from about 2 mol % to about 65 mol %, from about 5 mol % to about 65 mol %, from about 10 mol % to about 65 mol %, from about 10 mol % to about 55 mol %, from about 10 mol % to about 50 mol %, or any range therein.

[0526] In some embodiments, the proportion of phospholipid present in the lipid nanoparticle compositions is about 65 mol %, about 60 mol %, about 55 mol %, about 50 mol %, about 45 mol %, about 44 mol %, about 43 mol %, about 42 mol %, about 41 mol %, about 40 mol %, about 39 mol %, about 38 mol %, about 37 mol %, about 36 mol %, about 35 mol %, about 34 mol %, about 33 mol %, about 32 mol %, about 31 mol %, about 30 mol %, about 29 mol %, about 28 mol %, about 27 mol %, about 26 mol %, about 25 mol %, about 24 mol %, about 23 mol %, about 22 mol %, about 21 mol %, about 20 mol %, about 19 mol %, about 18 mol %, about 17 mol %, about 16 mol %, about 15 mol %, about 14 mol %, about 13 mol %, about 12 mol %, about 11 mol %, about 10 mol %, about 9 mol %, about 8 mol %, about 7 mol %, about 6 mol %, about 5 mol %, about 4 mol %, about 3 mol %, about 2 mol %, or any portion or fraction thereof.

[0527] Sterol

[0528] Lipid nanoparticle composition disclosed herein may include sterol and / or sterol derivatives. The term “sterol” as used herein include, but not limited to, cholesterol, sitosterol, fecosterol, ergosterol, campesterol, stigmasterol or their derivatives. In some embodiments, lipid nanoparticle composition comprises cholesterol and / or cholesterol derivatives. Non- limiting examples of cholesterol and cholesterol derivatives include 5 a- cholestanol, 5P-coprostanol, cholesteryl-(2’-hydroxy)-ethyl ether, cholesteryl- (4’- hydroxy)-butyl ether, 6-ketocholestanol, 5a-cholestane, cholestenone, 5a-cholestanone, 5P-cholestanone, cholesteryl decanoate, or mixtures thereof. Methods of making cholesterol and cholesterol derivatives are well known in the art.

[0529] The proportion of sterol present in the lipid nanoparticle compositions may be from about 20 mol % to about 65 mol % or any range therein.

[0530] In some embodiments, the proportion of sterol present in the lipid nanoparticle compositions is from about 20 mol % to about 65 mol %, from about 25 mol % to about 65 mol %, from about 30 mol % to about 65 mol %, from about 31 mol % to about 60 mol %, from about 32 mol % to about 60 mol %, from about 33 mol % to about 60 mol %, from about 34 mol % to about 60 mol %, from about 35 mol % to about 60 mol %, or any range therein. In some embodiments, the proportion of sterol present in the lipid nanoparticle compositions is about 65 mol %, about 60 mol %, about 55 mol %, about 50 mol %, about 45 mol %, about 44 mol %, about 43 mol %, about 42 mol %, about 41 mol %, about 40 mol %, about 39 mol %, about 38 mol %, about 37 mol %, about 36 mol %, about 35 mol %, about 34 mol %, about 33 mol %, about 32 mol %, about 31 mol %, about 30 mol %, about 29 mol %, about 28 mol %, about 27 mol %, about 26 mol %, about 25 mol %, about 24 mol %, about 23 mol %, about 22 mol %, about 21 mol %, about 20 mol %, or any portion or fraction thereof.

[0531] PEG-lipid

[0532] The term PEG-lipid, pegylated lipid, PEG linked lipid, PEG conjugated lipid, PEG- lipid conjugate, PEG modified lipid have been used interchangeably to mean polyethylene glycol linked to a lipid moiety. The lipid moiety may be linked directly to the PEG molecule or through a linker. In some embodiments, a PEG-lipid comprises a PEG- modified phosphatidylethanolamines, PEG-modified phosphatidic acids, PEG-modified ceramides, PEG-modified dialkylamines, PEG-modified diacylglycerols, PEG-modified dialkylglycerols, and / or PEG-modified cholesterol, and / or mixtures thereof. The methods of making PEG-lipid are well known to persons skilled in the art.

[0533] In some embodiments, PEG-lipid is selected from mPEG-Dimyristoyl glycerol (mPEG-DMG), mPEG-N,N-Ditetradecylacetamide (mPEG-DTA or ALC0159), mPEG- Cholesterol (mPEG-CLS), mPEG-DSPE, mPEG-DMPE, mPEG-DPPE, mPEG-DLPE, mPEG-DOPE, mPEG-DPPC, mPEG-DSPC, l,2-Distearoyl-sn-Glycero-3- Phosphoethanolamine with conjugated methoxyl poly(ethylene glycol) (mPEG-DSPE), l,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (mPEG2000-DMG), a- (3 ’ - { [ 1 ,2-di(myristyloxy)propanoxy]carbonylamino }propyl)-co-methoxy, polyoxyethylene (mPEG2000C-DMG)), or mixtures thereof.

[0534] The PEG moiety of the PEG-lipid may comprise an average molecular weight ranging from 0.5 kDa to 10 kDa. In some embodiments, the PEG-lipid has an average molecular weight of about 0.5 kDa to 5 kDa, about 0.5 kDa to 4 kDa, 0.5 kDa to 3 kDa, 0.5 kDa to 2 kDa. In preferred embodiments, the PEG-lipid has an average molecular weight of about 0.5 kDa to about 2 kDa.

[0535] The proportion of PEG-lipid present in the lipid nanoparticle compositions may be from about 0.2 mol % to about 2.0 mol % or any range therein. In some embodiments, the proportion of PEG-lipid present in the lipid nanoparticle compositions is from about 0.2 mol % to about 2.0 mol %, from about 0.2 mol % to about 1.9 mol %, from about 0.2 mol % to about 1.8 mol %, from about 0.2 mol % to about 1.7 mol %, from about 0.2 mol % to about 1.6 mol %, from about 0.2 mol % to about 1.5 mol %, or any range therein.

[0536] In some embodiments, the proportion of PEG-lipid present in the lipid nanoparticle compositions is about 0.2 mol %, about 0.3 mol %, about 0. 4 mol %, about 0.5 mol %, about 0.6 mol %, about 0.7 mol %, about 0.8 mol %, about 0.9 mol %, about 1.0 mol %, about 1.1 mol %, about 1.2 mol %, about 1.3 mol %, about 1.4 mol %, about 1.5 mol %, about 1.6 mol %, about 1.7 mol %, about 1.8 mol %, about 1.9 mol %, about 2.0 mol %, or any portion or fraction thereof.

[0537] In some embodiments, the lipid nanoparticle composition additionally contains an ionizable polymer.

[0538] Ionizable polymer

[0539] As used herein the term “polymer” means a compound formed from a plurality of repeating units called monomers. Polymers are produced through a process called polymerization wherein two or more monomers are linked through chemical bonds to form the polymer. In some embodiments, the polymer is branched or unbranched. In some embodiments, the polymer is homopolymer, i.e., comprising same type of repeat units or monomers, or heteropolymer, i.e., comprising more than one type of repeat units or monomers. The terms heteropolymer and copolymer have been used interchangeably herein.

[0540] The term “Ionizable polymer” as used herein means, a polymer that can exist in a positively charged or neutral form depending on the pH of the solution or environment, for example, ionizable polymer will be cationic (positively charged) when pH of the solution is below the pKa of the ionizable polymer and neutral (no charge) when pH of the solution is same or above the pKa of the ionizable polymer. In some embodiments, ionizable polymer is positively charge in acidic pH i.e., pH 1.0 to pH 6.9. In some embodiments, ionizable polymer is neutral (no charge) around physiological pH (pH 7.0 to pH 7.5).

[0541] In some embodiments, the ionizable polymer is a biocompatible polymer or biodegradable polymer. The term “biocompatible polymer” and “biodegradable polymer” have been used interchangeably to mean a polymer that is substantially free from any deleterious effects when introduced into a living or biological system. Such polymers are capable of undergoing degradation when introduced into the living or biological systems and are not expected to produce significant toxicity or immunological response.

[0542] In some embodiments, the lipid nanoparticle compositions comprise an ionizable polymer. The ionizable polymer may be selected from a chitosan, chitosan derivatives, cellulose derivatives, a poly-L-lysine (PLL), a protamine, a polyethyleneimine, their derivatives, or a combination thereof.

[0543] In some embodiments, the ionizable polymer is positively charged at acidic pH i.e., pH 1.0 to 6.9 and is neutral around physiological pH (pH 7.0 to 7.5).

[0544] The proportion of ionizable polymer present in the lipid nanoparticle compositions may be from about 1 mol % to about 25 mol %.

[0545] In some embodiments, the proportion of ionizable polymer present in the lipid nanoparticle compositions is from about 1 mol % to about 25 mol %, from about 1 mol % to about 24 mol %, from about 1 mol % to about 23 mol %, from about 1 mol % to about 22 mol %, from about 1 mol % to about 21 mol %, from about 1 mol % to about 20 mol %, from about 1 mol % to about 19 mol %, from about 1 mol % to about 18 mol %, from about 1 mol % to about 17 mol %, from about 1 mol % to about 16 mol %, from about 1 mol % to about 15 mol %, or any range therein.

[0546] In some embodiments, the proportion of ionizable polymer present in the lipid nanoparticle compositions is about 1 mol %, about 2 mol %, about 3 mol %, about 4 mol %, about 5 mol %, about 6 mol %, about 7 mol %, about 8 mol %, about 9 mol %, about 10 mol %, about 11 mol %, about 12 mol %, about 13 mol %, about 14 mol %, about 15 mol %, about 16 mol %, about 17 mol %, about 18 mol %, about 19 mol %, about 20 mol %, about 21 mol %, about 22 mol %, about 23 mol %, about 24 mol %, about 25 mol %, or any portion or fraction thereof.

[0547] In some embodiments, the preferred ionizable polymer comprises a chitosan, chitosan derivatives, cellulose derivatives, a poly-L-lysine (PLL), a protamine, a polyethyleneimine, and / or their derivatives or a combination thereof. Method of treatment

[0548] In some aspects, provided herein is a method of treating or preventing a disease, comprising administering to a subject in need thereof the multisubunit nucleic acid sequence as described herein.

[0549] In some aspects, provided herein is a method of treating or preventing a disease, comprising administering to a subject in need thereof the lipid nanoparticle composition comprising a cationic lipid, a phospholipid, a sterol, a PEG-lipid and the multisubunit nucleic acid sequence as described herein.

[0550] In some aspects, provided herein is a method of treating or preventing a disease, comprising administering to a subject in need thereof the lipid nanoparticle composition comprising a cationic lipid, a phospholipid, a sterol, a PEG-lipid and the multisubunit nucleic acid sequence as described herein, wherein the cationic lipid is represented by any one of formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), or a combination thereof.

[0551] In some aspects, provided herein is a method of treating or preventing a disease, comprising administering to a subject in need thereof the lipid nanoparticle composition comprising an ionizable polymer, a cationic lipid, a phospholipid, a sterol, a PEG-lipid and the multisubunit nucleic acid sequence as described herein.

[0552] In some aspects, provided herein is a method of treating or preventing a disease, comprising administering to a subject in need thereof the lipid nanoparticle composition comprising an ionizable polymer, a cationic lipid, a phospholipid, a sterol, a PEG-lipid and the multisubunit nucleic acid sequence as described herein, wherein the cationic lipid is represented by any one of formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), or a combination thereof.

[0553] In some aspects, provided herein is a method of treating or preventing a disease, comprising administering to a subject in need thereof a vaccine comprising the multisubunit nucleic acid as described herein.

[0554] In some aspects, provided herein is a method of treating or preventing a disease, comprising administering to a subject in need thereof a vaccine comprising the lipid nanoparticle composition, wherein the lipid nanoparticle composition comprises a cationic lipid, a phospholipid, a sterol, a PEG-lipid, and the multisubunit nucleic acid as described herein. In some aspects, provided herein is a method of treating or preventing a disease, comprising administering to a subject in need thereof a vaccine comprising the lipid nanoparticle composition, wherein the lipid nanoparticle composition comprises a cationic lipid, a phospholipid, a sterol, a PEG-lipid, and the multisubunit nucleic acid as described herein, wherein the cationic lipid is represented by any one of formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), or a combination thereof.

[0555] In some aspects, provided herein is a method of treating or preventing a disease, comprising administering to a subject in need thereof a vaccine comprising the lipid nanoparticle composition, wherein the lipid nanoparticle composition comprises an ionizable polymer, a cationic lipid, a phospholipid, a sterol, a PEG-lipid, and the multisubunit nucleic acid described herein.

[0556] In some aspects, provided herein is a method of treating or preventing a disease, comprising administering to a subject in need thereof a vaccine comprising the lipid nanoparticle composition, wherein the lipid nanoparticle composition comprises an ionizable polymer, a cationic lipid, a phospholipid, a sterol, a PEG-lipid, and the multisubunit nucleic acid described herein, wherein the cationic lipid is represented by any one of formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), or a combination thereof.

[0557] In some aspects, the disclosure relates to use of a multisubunit nucleic acid as described herein, in the manufacture of a medicament for the treatment or prevention of a disease in a subject.

[0558] In some aspects, the disclosure relates to use of a lipid nanoparticle composition comprising a cationic lipid, a phospholipid, a sterol, a PEG-lipid, and the multisubunit nucleic acid sequence as described herein, in the manufacture of a medicament for the treatment or prevention of a disease in a subject.

[0559] In some aspects, the disclosure relates to use of a lipid nanoparticle composition comprising a cationic lipid, a phospholipid, a sterol, a PEG-lipid, and the multisubunit nucleic acid sequence as described herein, in the manufacture of a medicament for the treatment or prevention of a disease in a subject, wherein the cationic lipid is represented by any one of formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), or a combination thereof. In some aspects, the disclosure relates to use of a lipid nanoparticle composition comprising an ionizable polymer, a cationic lipid, a phospholipid, a sterol, a PEG-lipid, and the multisubunit nucleic acid sequence as described herein, in the manufacture of a medicament for the treatment or prevention of a disease in a subject.

[0560] In some aspects, the disclosure relates to use of a lipid nanoparticle composition comprising an ionizable polymer, a cationic lipid, a phospholipid, a sterol, a PEG-lipid, and the multisubunit nucleic acid sequence as described herein, in the manufacture of a medicament for the treatment or prevention of a disease in a subject, wherein the cationic lipid is represented by any one of formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), or a combination thereof.

[0561] In some embodiments, the disclosure relates to use of a vaccine comprising the multisubunit nucleic acid as described herein, in the manufacture of a medicament for the treatment or prevention of a disease in a subject.

[0562] In some embodiments, the disclosure relates to use of a vaccine comprising a lipid nanoparticle composition, wherein the lipid nanoparticle composition comprises a cationic lipid, a phospholipid, a sterol, a PEG-lipid, and the multisubunit nucleic acid described herein, in the manufacture of a medicament for the treatment or prevention of a disease in a subject.

[0563] In some embodiments, the disclosure relates to use of a vaccine comprising a lipid nanoparticle composition, wherein the lipid nanoparticle composition comprises a cationic lipid, a phospholipid, a sterol, a PEG-lipid, and the multisubunit nucleic acid described herein, in the manufacture of a medicament for the treatment or prevention of a disease in a subject, wherein the cationic lipid is represented by any one of formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), or a combination thereof.

[0564] In some embodiments, the disclosure relates to use of a vaccine comprising a lipid nanoparticle composition, wherein the lipid nanoparticle composition comprises an ionizable polymer, a cationic lipid, a phospholipid, a sterol, a PEG-lipid, and the multisubunit nucleic acid described herein, in the manufacture of a medicament for the treatment or prevention of a disease in a subject.

[0565] In some embodiments, the disclosure relates to use of a vaccine comprising a lipid nanoparticle composition, wherein the lipid nanoparticle composition comprises an ionizable polymer, a cationic lipid, a phospholipid, a sterol, a PEG-lipid, and the multisubunit nucleic acid described herein, in the manufacture of a medicament for the treatment or prevention of a disease in a subject, wherein the cationic lipid is represented by any one of formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), or a combination thereof.

[0566] In some embodiments, the disease is cervical cancer, vaginal cancer, vulvar cancer, penile cancer, anal cancer, rectal cancer, head and neck cancer, lung cancer, skin cancer, or oropharyngeal cancer. In some embodiments, the disease is a skin infection (e.g., a noncancerous skin growth, a wart, or a verrucae). In some embodiments, the wart is a common wart, a plantar wart, a flat wart, a genital wart, or laryngeal papillomatosis.

[0567] In some embodiments, the multisubunit nucleic acid sequence is present in biologically effective amount or therapeutically effective amount. In some embodiments, the biologically effective amount of the multisubunit nucleic acid sequence is between 0.1 pg to 2000 pg, 0.1 pg to 1800 pg, 0.1 pg to 1600 pg, 0.1 pg to 1400 pg, 0.1 pg to 1200 pg, 0.1 pg to 1000 pg, 0.1 pg to 950 pg, 0.1 pg to 900 pg, 0.1 pg to 850 pg, 0.1 pg to 800 pg, 0.1 pg to 750 pg, 0.1 pg to 700 pg, 0.1 pg to 650 pg, 0.1 pg to 600 pg, 0.1 pg to 550 pg, 0.1 pg to 500 pg, 0.1 pg to 450 pg, 0.1 pg to 400 pg, 0.1 pg to 350 pg, 0.1 pg to 300 pg, 0.1 pg to 250 pg, 0.1 pg to 200 pg, 0.1 to 175 pg, 0.1 to 150 pg, 0.1 to 125 pg, 0.1 to

[0568] 100 pg, 0.1 pg to 90 pg, 0.1 pg to 80 pg, 0.1 pg to 70 pg, 0.1 pg to 60 pg, 0.1 pg to 50 pg,

[0569] 0.1 pg to 40 pg, 0.1 pg to 30 pg, 0.1 pg to 20 pg, 0.1 pg to 10 pg, 0.1 pg to 5 pg, or any range therein.

[0570] In some embodiments, the biologically effective amount of the multisubunit nucleic acid sequence is from about 0.1 pg to 1000 pg, 0.1 pg to 950 pg, 0.1 pg to 900 pg, 0.1 pg to 850 pg, 0.1 pg to 800 pg, 0.1 pg to 750 pg, 0.1 pg to 700 pg, 0.1 pg to 650 pg, 0.1 pg to 600 pg, 0.1 pg to 550 pg, 0.1 pg to 500 pg, or any range therein.

[0571] In some embodiments, the biologically effective amount of the multisubunit nucleic acid sequence is 0.1 pg, 0.2 pg, 0.3 pg, 0.4 pg, 0.5 pg, 0.6 pg, 0.7 pg, 0.8 pg, 0.9 pg, 1 pg, 2 pg, 3 pg, 4 pg, 5 pg, 6 pg, 7 pg, 8 pg, 9 pg, 10 pg, 15 pg, 20 pg, 25 pg, 30 pg, 35 pg, 40 pg, 45 pg, 50 pg, 55 pg, 60 pg, 65 pg, 70 pg, 75 pg, 80 pg, 85 pg, 90 pg, 95 pg, 100 pg, 110 pg, 120 pg, 130 pg, 140 pg, 150 pg, 160 pg, 170 pg, 180 pg, 190 pg, 200 pg, 220 pg, 240 pg, 260 pg, 280 pg, 300 pg, 350 pg, 400 pg, 450 pg, 500 pg, 600 pg, 700 pg, 800 pg, 900 pg, 1000 pg, 1100 pg, 1200 pg, 1300 pg, 1400 pg, 1500 pg, 1600 pg, 1700 pg, 1800 pg, 1900 pg, 2000 pg or any portion or fraction thereof. In another aspect, provided herein is a multisubunit nucleic acid comprising a plurality of self- assembling immunogenic sequences, wherein each self-assembling immunogenic sequence is connected to an adjacent self- assembling immunogenic sequence of the plurality by one or more cleavage sequence, wherein the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the self-assembling immunogenic sequence of the plurality, and wherein the self-assembling immunogenic sequence is obtained or derived from LI protein of a papillomavirus.

[0572] In another aspect, provided herein is a multisubunit nucleic acid comprising a plurality of self-assembling immunogenic sequences, wherein the plurality of selfassembling immunogenic sequences further comprises one or more target sequence, wherein each self-assembling immunogenic sequence is connected to an adjacent selfassembling immunogenic sequence of the plurality by one or more cleavage sequence, wherein the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the self-assembling immunogenic sequence of the plurality, wherein the self-assembling immunogenic sequence is obtained or derived from LI protein of a papillomavirus and the target sequence is obtained or derived from L2 protein, El protein, E2 protein, E4 protein, E5 protein, E6 protein, E7 protein, B cell epitope, T cell epitope, or a combination thereof of a papillomavirus.

[0573] In one aspect, provided herein is a multisubunit nucleic acid comprising a plurality of plurality of self-assembling immunogenic sequences, wherein the multisubunit nucleic acid comprises either a plurality of self-assembling immunogenic sequences or a plurality of self-assembling immunogenic sequences each further comprising one or more target sequence, or a combination thereof, wherein each self- assembling immunogenic sequence is connected to an adjacent self-assembling immunogenic sequence of the plurality by one or more cleavage sequence, and wherein the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the self-assembling immunogenic sequence of the plurality, wherein the self-assembling immunogenic sequence is obtained or derived from LI protein of a papillomavirus and the target sequence is obtained or derived from L2 protein, El protein, E2 protein, E4 protein, E5 protein, E6 protein, E7 protein, B cell epitope, T cell epitope, or a combination thereof of a papillomavirus.

[0574] In one aspect, provided herein is a multisubunit nucleic acid comprising a plurality of polynucleotide sequences, wherein some or all polynucleotide sequences of the plurality comprises either a self-assembling immunogenic sequence, or a self-assembling immunogenic sequence further comprising one or more target sequence, or a combination thereof, wherein each polynucleotide sequence of the plurality is connected to an adjacent polynucleotide sequence of the plurality by one or more cleavage sequence, wherein the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the polynucleotide sequence of the plurality, wherein the self-assembling immunogenic sequence is obtained or derived from LI protein of a papillomavirus and the target sequence is obtained or derived from L2 protein, El protein, E2 protein, E4 protein, E5 protein, E6 protein, E7 protein, B cell epitope, T cell epitope, or a combination thereof of a papillomavirus.

[0575] In another aspect, provided herein is a multisubunit nucleic acid comprising a plurality of polynucleotide sequences, wherein one polynucleotide sequence of the plurality comprises a self-assembling immunogenic sequence and another polynucleotide sequence of the plurality comprises a self-assembling immunogenic sequence further comprising one or more target sequence, or a combination thereof, wherein each polynucleotide sequence of the plurality is connected to an adjacent polynucleotide sequence of the plurality by one or more cleavage sequence, wherein the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the polynucleotide sequence of the plurality, wherein the self-assembling immunogenic sequence is obtained or derived from LI protein of a papillomavirus and the target sequence is obtained or derived from L2 protein, El protein, E2 protein, E4 protein, E5 protein, E6 protein, E7 protein, B cell epitope, T cell epitope, or a combination thereof of a papillomavirus.

[0576] Embodiments:

[0577] Some of the embodiments of the present disclosure, set out in the following numbered paragraphs, are:

[0578] Embodiment set 1

[0579] 1. A multisubunit nucleic acid comprising either a plurality of self-assembling immunogenic sequences or a plurality of self-assembling immunogenic sequences each further comprising one or more target sequence, or a combination thereof, wherein each self-assembling immunogenic sequence is connected to an adjacent self-assembling immunogenic sequence of the plurality by one or more cleavage sequence, and wherein the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the self-assembling immunogenic sequences of the plurality.

[0580] 2. A multisubunit nucleic acid comprising a plurality of self-assembling immunogenic sequences, wherein each self-assembling immunogenic sequence is connected to an adjacent self-assembling immunogenic sequence of the plurality by one or more cleavage sequence, and wherein the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the self-assembling immunogenic sequences of the plurality.

[0581] 3. The multisubunit nucleic acid according to paragraph 2, wherein some or all of the self-assembling immunogenic sequences of the plurality further comprises one or more target sequence, wherein each self- assembling immunogenic sequence of the plurality is connected to an adjacent self-assembling immunogenic sequence of the plurality by one or more cleavage sequence, and wherein the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the self-assembling immunogenic sequences of the plurality.

[0582] 4. A multisubunit nucleic acid comprising a plurality of self-assembling immunogenic sequences, wherein each self-assembling immunogenic sequence further comprises one or more target sequence, wherein each self-assembling immunogenic sequence of the plurality is connected to an adjacent self-assembling immunogenic sequence of the plurality by one or more cleavage sequence, and wherein the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the selfassembling immunogenic sequences of the plurality.

[0583] 5. A multisubunit nucleic acid encoding a multisubunit peptide, wherein the multisubunit peptide comprises either a plurality of self-assembling immunogenic peptides or a plurality of self-assembling immunogenic peptide further comprising one or more target peptide, or a combination thereof, wherein each self-assembling immunogenic peptide is connected to an adjacent self-assembling immunogenic peptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus of one or more of the self-assembling immunogenic peptides of the plurality.

[0584] 6. A multisubunit nucleic acid encoding a multisubunit peptide, wherein the multisubunit peptide comprises a plurality of self-assembling immunogenic peptides, wherein each self-assembling immunogenic peptide of the plurality is connected to an adjacent self-assembling immunogenic peptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus of one or more of the self- assembling immunogenic peptides of the plurality.

[0585] 7. The multisubunit nucleic acid according to paragraph 6, wherein some or all of the self-assembling immunogenic peptides of the plurality further comprises one or more target peptide, wherein each self-assembling immunogenic peptide of the plurality is connected to an adjacent self-assembling immunogenic peptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus of one or more of the self-assembling immunogenic peptides of the plurality.

[0586] 8. A multisubunit nucleic acid encoding a multisubunit peptide, wherein the multisubunit peptide comprises a plurality of self-assembling immunogenic peptides further comprising one or more target peptide, wherein each self-assembling immunogenic peptide of the plurality is connected to an adjacent self-assembling immunogenic peptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus of one or more of the self-assembling immunogenic peptides of the plurality.

[0587] 9. The multisubunit nucleic acid according to any one of the preceding paragraphs, wherein total number of the self-assembling immunogenic sequences or the selfassembling immunogenic peptides are not more than 100. 10. The multisubunit nucleic acid according to paragraph 9, wherein total number of the self-assembling immunogenic sequences or the self-assembling immunogenic peptides are between 2-5, 2-10, 10-20, 20-30, 30-40, 40-50, 50-60, 60-70, 70-80, 80-90, or 90-99.

[0588] 11. The multisubunit nucleic acid according to any one of the preceding paragraphs, wherein the multisubunit nucleic acid is a DNA or an RNA.

[0589] 12. The multisubunit nucleic acid according to paragraph 11, wherein the RNA is an mRNA.

[0590] 13. The multisubunit nucleic acid according to paragraph 12, wherein, the mRNA is 0.5 kb to 1 kb, 1 kb to 2 kb, 2 kb to 3 kb, 3 kb to 4 kb, 4 kb to 5 kb, 5 kb to 6 kb, 6 kb to 7 kb, 7 kb to 8 kb, 8 kb to 9 kb, 9 kb to 10 kb, 10 kb to 11 kb, 11 kb to 12 kb, 12 kb to 13 kb, 13 kb to 14 kb, 14 kb to 15 kb, 15 kb to 16 kb, 16 kb to 17 kb, 17 kb to 18 kb, 18 kb to 19 kb, 19 kb to 20 kb in length, or any range therein.

[0591] 14. The multisubunit nucleic acid according to any one of the paragraphs 12-13, wherein the mRNA is obtained through a single IVT process or step.

[0592] 15. The multisubunit nucleic acid according to any one of the paragraphs 1, 3-4, and 9- 14, wherein the target sequence comprises a linker sequence on the 5’(5-prime), 3’ (3- prime) or on both sides of the target sequence.

[0593] 16. The multisubunit nucleic acid according to paragraph 15, wherein the linker sequence encodes a linker peptide.

[0594] 17. The multisubunit nucleic acid according to any one of the paragraphs 5 and 7-8, wherein the target peptide comprises a linker peptide on the C-terminus, N-terminus, or on both ends of the target peptide.

[0595] 18. The multisubunit nucleic acid according to any one of the paragraphs 16-17, wherein the linker peptide is an amino acid linker, a foldon, a scaffold, or a combination thereof. 19. The multisubunit nucleic acid according to paragraph 18, wherein the linker peptide is the amino acid linker.

[0596] 20. The multisubunit nucleic acid according to paragraph 19, wherein the amino acid linker comprises 2 to 49 amino acids.

[0597] 21. The multisubunit nucleic acid according to any one of the paragraphs 19-20, wherein the amino acid linker is a glycine serine linker, a glycine proline linker, a glycine threonine linker, an alanine serine linker, any combination of two amino acids, or a combination thereof.

[0598] 22. The multisubunit nucleic acid according to paragraph 18, wherein the linker peptide is the foldon.

[0599] 23. The multisubunit nucleic acid according to paragraph 18, wherein the linker peptide is the scaffold.

[0600] 24. The multisubunit nucleic acid according to any one of the paragraphs 18-21, wherein the linker peptide comprises the amino acid linker and the foldon.

[0601] 25. The multisubunit nucleic acid according to any one of the paragraphs 18-21, wherein the linker peptide comprises the amino acid linker and the scaffold.

[0602] 26. The multisubunit nucleic acid according to any one of the paragraphs 18-21, wherein the linker peptide comprises the amino acid linker, the foldon, and the scaffold.

[0603] 27. The multisubunit nucleic acid according to paragraph 18, wherein the linker peptide comprises the foldon and the scaffold.

[0604] 28. The multisubunit nucleic acid according to any one of the paragraphs 16-27, wherein the linker peptide has an amino acid sequence of any one of SEQ ID NOs: 1-40. 29. The multisubunit nucleic acid according to any one of the paragraphs 1-4 and 9-28, wherein the one or more cleavage sequence encodes one or more cleavage peptides.

[0605] 30. The multisubunit nucleic acid according to any one of the paragraphs 5-29, wherein the one or more cleavage peptides are optionally connected to each other by a linker peptide.

[0606] 31. The multisubunit nucleic acid according to paragraph 30, wherein the cleavage peptide is a golgi specific cleavage peptide, a self cleaving peptide, or a combination thereof.

[0607] 32. The multisubunit nucleic acid according to any one of the paragraphs 5-31, wherein the cleavage peptide has an amino acid sequence of any one of SEQ ID NOs: 41-55.

[0608] 33. The multisubunit nucleic acid according to any one of the paragraphs 1-4 and 9-32, wherein the one or more signal sequence encodes one or more signal peptides.

[0609] 34. The multisubunit nucleic acid according to any one of the paragraphs 5-33, wherein the one or more signal peptides are optionally connected to each other by a linker peptide.

[0610] 35. The multisubunit nucleic acid according to any one of the paragraphs 33-34, wherein the signal peptide is present on the amino-terminus of one or more of the selfassembling immunogenic peptides of the plurality.

[0611] 36. The multisubunit nucleic acid according to any one of the paragraphs 5-35, wherein the signal peptide has an amino acid sequence of any one of SEQ ID NOs: 56-75.

[0612] 37. The multisubunit nucleic acid according to any one of the paragraphs 1-4, wherein the self- assembling immunogenic sequence encodes a self-assembling immunogenic peptide. 38. The multisubunit nucleic acid according to any one of the paragraphs 5-37, wherein the self-assembling immunogenic peptide is obtained or derived from LI protein of a papillomavirus.

[0613] 39. The multisubunit nucleic acid according to paragraph 38, wherein the LI protein comprises a mutation at: a) a codon encoding an amino acid C at the position corresponding to the position 175 of SEQ ID NO:712, wherein the mutation is a substitution replacing the amino acid C with an amino acid A, b) a codon encoding an amino acid C at the position corresponding to the position 428 of SEQ ID NO:712, wherein the mutation is a substitution replacing the amino acid C with an amino acid A, or c) a combination thereof.

[0614] 40. The multisubunit nucleic acid according to any one of the paragraphs 1 and 3-4, wherein the target sequence encodes a target peptide.

[0615] 41. The multisubunit nucleic acid according to any one of the paragraphs 5 and 7-40, wherein the target peptide is obtained or derived from L2 protein, El protein, E2 protein, E4 protein, E5 protein, E6 protein, E7 protein, B cell epitope, T cell epitope, or a combination thereof of a papillomavirus.

[0616] 42. The multisubunit nucleic acid according to paragraph 41, wherein the target peptide is inserted into the BC loop, DE loop, EF loop, FG loop, HI loop, or a combination thereof of the LI protein of a papillomavirus.

[0617] 43. The multisubunit nucleic acid according to paragraph 42, wherein the target peptide is inserted into the DE loop, FG loop, or a combination thereof of the LI protein of a papillomavirus.

[0618] 44. The multisubunit nucleic acid according to any one of the paragraphs 38-43, wherein the papillomavirus is selected from the group comprising bovine papillomavirus (BPV), canine oral papillomavirus (COPY), cotton tail rabbit papillomavirus (CRPV), european elk papillomavirus (EEPV), rhesus monkey papillomavirus (RhPV), human papillomavirus (HPV), or a combination thereof.

[0619] 45. The multisubunit nucleic acid according to paragraph 44, wherein the human papillomavirus (HPV) is selected from the group comprising HPV1, HPV2, HPV3, HPV4, HPV5, HPV6, HPV7, HPV8, HPV9, HPV10, HPV11, HPV12, HPV13, HPV14, HPV15, HPV16, HPV17, HPV18, HPV19, HPV20, HPV21, HPV22, HPV23, HPV24, HPV25, HPV26, HPV27, HPV28, HPV29, HPV30, HPV31, HPV32, HPV33, HPV34, HPV35, HPV36, HPV37, HPV38, HPV39, HPV40, HPV41, HPV42, HPV43, HPV44, HPV45, HPV47, HPV48, HPV49, HPV50, HPV51, HPV52, HPV53, HPV54, HPV56, HPV57, HPV58, HPV59, HPV60, HPV61, HPV62, HPV63, HPV65, HPV66, HPV67, HPV68, HPV69, HPV70, HPV71, HPV72, HPV73, HPV74, HPV75, HPV76, HPV77, HPV78, HPV80, HPV81, HPV82, HPV83, HPV84, HPV85, HPV86, HPV87, HPV88, HPV89, HPV90, HPV91, HPV92, HPV93, HPV94, HPV95, HPV96, HPV97, HPV98, HPV99, HPV100, HPV101, HPV102, HPV103, HPV104, HPV105, HPV106, HPV107, HPV108, HPV109, HPV110, HPV111, HPV112, HPV113, HPV114, HPV115, HPV116, HPV117, HPV118, HPV119, HPV120, HPV121, HPV122, HPV123, HPV124, HPV125, HPV126, HPV127, HPV128, HPV129, HPV130, HPV131, HPV132, HPV133, HPV134, HPV135, HPV136, HPV137, HPV138, HPV139, HPV140, HPV141, HPV142, HPV143, HPV144, HPV145, HPV146, HPV147, HPV148, HPV148, HPV150, HPV151, HPV152, HPV153, HPV154, HPV155, HPV156, HPV157, HPV158, HPV159, HPV160, HPV161, HPV162, HPV163, HPV164, HPV165, HPV166, HPV167, HPV168, HPV169, HPV170, HPV171, HPV172, HPV173, HPV174, HPV175, HPV176, HPV177, HPV178, HPV179, HPV180, HPV181, HPV182, HPV183, HPV184, HPV185, HPV186, HPV187, HPV188, HPV189, HPV190, HPV191, HPV192, HPV193, HPV194, HPV195, HPV196, HPV197, HPV198, HPV199, HPV200, HPV201, HPV202, HPV203, HPV204, HPV205, HPV206, HPV207, HPV208, HPV209, HPV210, HPV211, HPV212, HPV213, HPV214, HPV215, HPV216, HPV217, HPV218, HPV219, HPV220, HPV221, HPV222, or a combination thereof.

[0620] 46. The multisubunit nucleic acid according to paragraph 45, wherein the human papillomavirus is selected from the group comprising HPV5, HPV6, HPV8, HPV11, HPV16, HPV18, HPV26, HPV30, HPV31, HPV33, HPV34, HPV35, HPV39, HPV42, HPV43, HPV44, HPV45, HPV51, HPV52, HPV53, HPV56, HPV58, HPV59, HPV66, HPV67, HPV68, HPV69, HPV70, HPV73, HPV82, HPV85, HPV97, or a combination thereof.

[0621] 47. The multisubunit nucleic acid according to any one of the paragraphs 38-46, wherein the LI protein has an amino acid sequence of any one of SEQ ID NOs: 76-1129 or 9409-9444.

[0622] 48. The multisubunit nucleic acid according to any one of the paragraphs 41-47, wherein the L2 protein has an amino acid sequence according to any one of SEQ ID NOs: 1130-3675.

[0623] 49. The multisubunit nucleic acid according to any one of the paragraphs 41-47, wherein the El protein has an amino acid sequence according to any one of SEQ ID NOs: 3676-4768.

[0624] 50. The multisubunit nucleic acid according to any one of the paragraphs 41-47, wherein the E2 protein has an amino acid sequence according to any one of SEQ ID NOs: 4769-5850.

[0625] 51. The multisubunit nucleic acid according to any one of the paragraphs 41-47, wherein the E4 protein has an amino acid sequence according to any one of SEQ ID NOs: 5851-6415.

[0626] 52. The multisubunit nucleic acid according to any one of the paragraphs 41-47, wherein the E5 protein has an amino acid sequence according to any one of SEQ ID NOs: 6416-6713.

[0627] 53. The multisubunit nucleic acid according to any one of the paragraphs 41-47, wherein the E6 protein has an amino acid sequence according to any one of SEQ ID NOs: 6714-7420. 54. The multisubunit nucleic acid according to any one of the paragraphs 41-47, wherein the E7 protein has an amino acid sequence according to any one of SEQ ID NOs: 7421-8044.

[0628] 55. The multisubunit nucleic acid according to any one of the paragraphs 41-47, wherein the B cell epitope has an amino acid sequence according any of SEQ ID NOs: 8045-8654.

[0629] 56. The multisubunit nucleic acid according to any one of the paragraphs 41-47, wherein the T cell epitope has an amino acid sequence according to any of SEQ ID NOs: 8655-9408.

[0630] 57. A lipid nanoparticle composition comprising a cationic lipid, a phospholipid, a sterol, a PEG-lipid, and the multisubunit nucleic acid according to any one of the preceding paragraphs.

[0631] 58. The lipid nanoparticle composition according to paragraph 57, wherein the cationic lipid is present in an amount from 10 mol percent to 70 mol percent.

[0632] 59. The lipid nanoparticle composition according to paragraph 57, wherein the phospholipid is present in an amount from 2 mol percent to 65 mol percent.

[0633] 60. The lipid nanoparticle composition according to paragraph 57, wherein the sterol is present in an amount from 20 mol percent to 65 mol percent.

[0634] 61. The lipid nanoparticle composition according to paragraph 57, wherein the PEG- lipid is present in an amount from 0.2 mol percent to 2.0 mol percent.

[0635] 62. The lipid nanoparticle composition according to paragraph 57, additionally comprising an ionizable polymer.

[0636] 63. The lipid nanoparticle composition according to paragraph 62, wherein the ionizable polymer is present in an amount from 1 mol percent to 25 mol percent. 64. The lipid nanoparticle composition according to any one of the paragraphs 57-63, wherein the cationic lipid is represented by any one of formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), or SM-102, or ALC-0315, or a combination thereof.

[0637] 65. The lipid nanoparticle composition according to any one of the paragraphs 57-64, wherein the cationic lipid is represented by formula (I).

[0638] 66. The lipid nanoparticle composition according to any one of the paragraphs 57-64, wherein the cationic lipid is represented by formula (II).

[0639] 67. The lipid nanoparticle composition according to any one of the paragraphs 57-64, wherein the cationic lipid is represented by formula (III).

[0640] 68. The lipid nanoparticle composition according to any one of the paragraphs 57-64, wherein the cationic lipid is represented by formula (IV).

[0641] 69. The lipid nanoparticle composition according to any one of the paragraphs 57-64, wherein the cationic lipid is represented by formula (V).

[0642] 70. The lipid nanoparticle composition according to any one of the paragraphs 57-64, wherein the cationic lipid is represented by formula (VI).

[0643] 71. The lipid nanoparticle composition according to any one of the paragraphs 57-64, wherein the cationic lipid is represented by formula (VII).

[0644] 72. The lipid nanoparticle composition according to any one of the paragraphs 57-64, wherein the cationic lipid is represented by formula (VIII).

[0645] 73. A multisubunit peptide encoded by the multisubunit nucleic acid according to any one of the paragraphs 1-56. 74. A multisubunit peptide comprising either a plurality of self-assembling immunogenic peptides or a plurality of self-assembling immunogenic peptides each further comprising one or more target peptide, or a combination thereof, wherein self-assembling immunogenic peptide is connected to an adjacent self-assembling immunogenic peptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus of one or more of the selfassembling immunogenic peptides of the plurality.

[0646] 75. A multisubunit peptide comprising a plurality of self-assembling immunogenic peptides, wherein each self-assembling immunogenic peptide of the plurality is connected to an adjacent self-assembling immunogenic peptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus of one or more of the self-assembling immunogenic peptides of the plurality.

[0647] 76. The multisubunit peptide according to paragraph 75, wherein some or all of the self-assembling immunogenic peptides of the plurality further comprises one or more target peptide, wherein each self-assembling immunogenic peptide of the plurality is connected to an adjacent self-assembling immunogenic peptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus of one or more of the self-assembling immunogenic peptides of the plurality.

[0648] 77. A multisubunit peptide comprising a plurality of self-assembling immunogenic peptides, wherein the plurality of self-assembling immunogenic peptides further comprises one or more target peptide, wherein each self-assembling immunogenic peptide of the plurality is connected to an adjacent self-assembling immunogenic peptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus of one or more of the self-assembling immunogenic peptides of the plurality.

[0649] 78. The multisubunit peptide according to any one of the paragraphs 74-77, wherein total number of the self-assembling immunogenic peptides are not more than 100. 79. The multisubunit peptide according to paragraph 78, wherein total number of the self-assembling immunogenic peptides are between 2-5, 2-10, 10-20, 20-30, 30-40, 40-50, 50-60, 60-70, 70-80, 80-90, or 90-99.

[0650] 80. The multisubunit peptide according to any one of the paragraphs 74 and 76-79, wherein the target peptide comprises a linker peptide on the C-terminus, N-terminus or both sides of the target peptide.

[0651] 81. The multisubunit peptide according to paragraph 80, wherein the linker peptide is an amino acid linker, a foldon, a scaffold, or a combination thereof.

[0652] 82. The multisubunit peptide according to paragraph 81, wherein the linker peptide is the amino acid linker.

[0653] 83. The multisubunit peptide according to paragraph 82, wherein the amino acid linker comprises 2 to 49 amino acids.

[0654] 84. The multisubunit peptide according to any one of the paragraphs 82-83, wherein the amino acid linker is a glycine serine linker, a glycine proline linker, a glycine threonine linker, an alanine serine linker, any combination of two amino acids, or a combination thereof.

[0655] 85. The multisubunit peptide according to paragraph 81, wherein the linker peptide is the foldon.

[0656] 86. The multisubunit peptide according to paragraph 81, wherein the linker peptide is the scaffold.

[0657] 87. The multisubunit peptide according to any one of the paragraphs 81-84, wherein the linker peptide comprises the amino acid linker and the foldon. 88. The multisubunit peptide according to any one of the paragraphs 81-84, wherein the linker peptide comprises the amino acid linker and the scaffold.

[0658] 89. The multisubunit peptide according to any one of the paragraphs 81-84, wherein the linker peptide comprises the amino acid linker, the foldon, and the scaffold.

[0659] 90. The multisubunit peptide according to paragraph 81 , wherein the linker peptide comprises the foldon and the scaffold.

[0660] 91. The multisubunit peptide according to any one of the paragraphs 80-90, wherein the linker peptide has an amino acid sequence of any one of SEQ ID NOs: 1-40.

[0661] 92. The multisubunit peptide according to any one of the paragraphs 74-91, wherein the one or more cleavage peptides are optionally connected to each other by a linker peptide.

[0662] 93. The multisubunit peptide according to paragraph 92, wherein the cleavage peptide is a golgi specific cleavage peptide, a self-cleaving peptide, or a combination thereof.

[0663] 94. The multisubunit peptide according to paragraph 93, wherein the cleavage peptide has an amino acid sequence of any one of SEQ ID NOs: 41-55.

[0664] 95. The multisubunit peptide according to any one of the paragraphs 74-94, wherein the signal peptide is present on the amino-terminus of one or more of the self-assembling immunogenic peptides of the plurality.

[0665] 96. The multisubunit peptide according to paragraph 95, wherein a second signal peptide is present on the amino-terminus of one or more self- assembling immunogenic peptides of the plurality.

[0666] 97. The multisubunit peptide according to any one of the paragraphs 95-96, wherein the signal peptide has an amino acid sequence of any one of SEQ ID NOs: 56-75. 98. The multisubunit peptide according to any one of the paragraphs 74-97, wherein the self-assembling immunogenic peptide is obtained or derived from LI protein of a papillomavirus.

[0667] 99. The multisubunit peptide according to paragraph 98, wherein the LI protein comprises a mutation at: a) the position corresponding to the position 175 of SEQ ID NO:712, wherein the mutation is a substitution replacing the amino acid C with an amino acid A, b) the position corresponding to the position 428 of SEQ ID NO:712, wherein the mutation is a substitution replacing the amino acid C with an amino acid A, or c) a combination thereof.

[0668] 100. The multisubunit peptide according to any one of the paragraphs 74 and 76-99, wherein the target peptide is obtained or derived from L2 protein, El protein, E2 protein, E4 protein, E5 protein, E6 protein, E7 protein, B cell epitope, T cell epitope, or a combination thereof of a papillomavirus.

[0669] 101. The multisubunit peptide according to paragraph 100, wherein the target peptide is inserted into the BC loop, DE loop, EF loop, FG loop, HI loop, or combination thereof of the LI protein of a papillomavirus.

[0670] 102. The multisubunit peptide according to paragraph 101, wherein the target peptide is inserted into the DE loop, the FG loop, or combination thereof of the LI protein of a papillomavirus.

[0671] 103. The multisubunit peptide according to any one of the paragraphs 98-102, wherein the papillomavirus is selected from the group comprising bovine papillomavirus (BPV), canine oral papillomavirus (COPV), cotton tail rabbit papillomavirus (CRPV), european elk papillomavirus (EEPV), rhesus monkey papillomavirus (RhPV), human papillomavirus (HPV), or a combination thereof.

[0672] 104. The multisubunit peptide according to paragraph 103, wherein the human papillomavirus (HPV) is selected from the group comprising HPV1, HPV2, HPV3, HPV4, HPV5, HPV6, HPV7, HPV8, HPV9, HPV10, HPV11, HPV12, HPV13, HPV14, HPV15, HPV16, HPV17, HPV18, HPV19, HPV20, HPV21, HPV22, HPV23, HPV24, HPV25, HPV26, HPV27, HPV28, HPV29, HPV3O, HPV31, HPV32, HPV33, HPV34, HPV35, HPV36, HPV37, HPV38, HPV39, HPV40, HPV41, HPV42, HPV43, HPV44, HPV45, HPV47, HPV48, HPV49, HPV5O, HPV51, HPV52, HPV53, HPV54, HPV56, HPV57, HPV58, HPV59, HPV6O, HPV61, HPV62, HPV63, HPV65, HPV66, HPV67, HPV68, HPV69, HPV70, HPV71, HPV72, HPV73, HPV74, HPV75, HPV76, HPV77, HPV78, HPV8O, HPV81, HPV82, HPV83, HPV84, HPV85, HPV86, HPV87, HPV88, HPV89, HPV90, HPV91, HPV92, HPV93, HPV94, HPV95, HPV96, HPV97, HPV98, HPV99, HPV1OO, HPV1O1, HPV102, HPV103, HPV104, HPV105, HPV106, HPV107, HPV108, HPV109, HPV11O, HPV111, HPV112, HPV113, HPV114, HPV115, HPV116, HPV117, HPV118, HPV119, HPV120, HPV121, HPV122, HPV123, HPV124, HPV125, HPV126, HPV127, HPV128, HPV129, HPV13O, HPV131, HPV132, HPV133, HPV134, HPV135, HPV136, HPV137, HPV138, HPV139, HPV140, HPV141, HPV142, HPV143, HPV144, HPV145, HPV146, HPV147, HPV148, HPV148, HPV15O, HPV151, HPV152, HPV153, HPV154, HPV155, HPV156, HPV157, HPV158, HPV159, HPV16O, HPV161, HPV162, HPV163, HPV164, HPV165, HPV166, HPV167, HPV168, HPV169, HPV170, HPV171, HPV172, HPV173, HPV174, HPV175, HPV176, HPV177, HPV178, HPV179, HPV18O, HPV181, HPV182, HPV183, HPV184, HPV185, HPV186, HPV187, HPV188, HPV189, HPV190, HPV191, HPV192, HPV193, HPV194, HPV195, HPV196, HPV197, HPV198, HPV199, HPV200, HPV201, HPV202, HPV203, HPV204, HPV205, HPV206, HPV207, HPV2O8, HPV209, HPV210, HPV211, HPV212, HPV213, HPV214, HPV215, HPV216, HPV217, HPV218, HPV219, HPV220, HPV221, HPV222, or a combination thereof.

[0673] 105. The multisubunit peptide according to paragraph 104, wherein the human papillomavirus is selected from the group comprising HPV5, HPV6, HPV8, HPV11, HPV16, HPV18, HPV26, HPV30, HPV31, HPV33, HPV34, HPV35, HPV39, HPV42, HPV43, HPV44, HPV45, HPV51, HPV52, HPV53, HPV56, HPV58, HPV59, HPV66, HPV67, HPV68, HPV69, HPV70, HPV73, HPV82, HPV85, HPV97, or a combination thereof. 106. The multisubunit peptide according to any one of the paragraphs 98-105, wherein the LI protein has an amino acid sequence of any one of SEQ ID NOs: 76-1129 or 9409- 9444.

[0674] 107. The multisubunit peptide according to any one of the paragraphs 100-106, wherein the L2 protein has an amino acid sequence according to any of SEQ ID NOs: 1130-3675.

[0675] 108. The multisubunit peptide according to any one of the paragraphs 100-106, wherein the El protein has an amino acid sequence according to any of SEQ ID NOs: 3676-4768.

[0676] 109. The multisubunit peptide according to any one of the paragraphs 100-106, wherein the E2 protein has an amino acid sequence according to any of SEQ ID NOs: 4769-5850.

[0677] 110. The multisubunit peptide according to any one of the paragraphs 100-106, wherein the E4 protein has an amino acid sequence according to any of SEQ ID NOs: 5851-6415.

[0678] 111. The multisubunit peptide according to any one of the paragraphs 100-106, wherein the E5 protein has an amino acid sequence according to any of SEQ ID NOs: 6416-6713.

[0679] 112. The multisubunit peptide according to any one of the paragraphs 100-106, wherein the E6 protein has an amino acid sequence according to any of SEQ ID NOs: 6714-7420.

[0680] 113. The multisubunit peptide according to any one of the paragraphs 100-106, wherein the E7 protein has an amino acid sequence according to any of SEQ ID NOs: 7421-8044.

[0681] 114. The multisubunit peptide according to any one of the paragraphs 100-106, wherein the B cell epitope has an amino acid sequence according any of SEQ ID NOs: 8045-8654.

[0682] 115. The multisubunit peptide according to any one of the paragraphs 100-106, wherein the T cell epitope has an amino acid sequence according to any of SEQ ID NOs: 8655- 9408. 116. A peptide nanoparticle comprising at least 2 or up to 500 self-assembling immunogenic peptides according to any one of the paragraphs 74-115.

[0683] 117. The peptide nanoparticle according to paragraph 116, comprising a homologous self-assembling immunogenic peptide, a heterologous self- assembling immunogenic peptide, an oligomeric complex, or a combination thereof.

[0684] 118. The peptide nanoparticle according to any one of the paragraphs 116-117, wherein the peptide nanoparticle is icosahedral, helical, spherical, rod-like, or a combination thereof.

[0685] 119. A vaccine comprising the multisubunit nucleic acid according to any one of the paragraphs 1-56.

[0686] 120. A vaccine comprising the lipid nanoparticle composition according to any one of the paragraphs 57-72.

[0687] 121. A vaccine comprising the peptide nanoparticle according to any one of the paragraphs 116-118.

[0688] 122. A method of treating or preventing a disease, comprising administering to a subject in need thereof the multisubunit nucleic acid according to any one of the paragraphs 1-56 or the vaccine according to any one of the paragraphs 119 or 121.

[0689] 123. A method of treating or preventing a disease, comprising administering to a subject in need thereof the lipid nanoparticle composition according to any one of the paragraphs 57-72 or the vaccine according to paragraph 120.

[0690] 124. The method according to any one of the paragraphs 122-123, wherein the disease is cervical cancer, vaginal cancer, vulvar cancer, penile cancer, anal cancer, rectal cancer, head and neck cancer, lung cancer, skin cancer, oropharyngeal cancer, or a skin infection (e.g., a noncancerous skin growth, a wart, or a verrucae). 125. Use of the multisubunit nucleic acid according to any one of the paragraphs 1-56 or the vaccine according to any one of the paragraphs 119 or 121, in the manufacture of a medicament for the treatment or prevention of a disease in a subject.

[0691] 126. Use of the lipid nanoparticle composition according to any one of the paragraphs 57-72 or the vaccine according to paragraph 120, in the manufacture of a medicament for the treatment or prevention of a disease in a subject.

[0692] 127. The use according to any one of the paragraphs 125-126, wherein the disease is cervical cancer, vaginal cancer, vulvar cancer, penile cancer, anal cancer, rectal cancer, head and neck cancer, lung cancer, skin cancer, oropharyngeal cancer, or a skin infection (e.g., a noncancerous skin growth, a wart, or a verrucae).

[0693] Embodiment set 2:

[0694] 1. A multisubunit nucleic acid comprising a plurality of polynucleotide sequences, wherein some or all polynucleotide sequences of the plurality comprises either a self-assembling immunogenic sequence, or a self-assembling immunogenic sequence further comprising one or more target sequence, or a combination thereof, wherein each polynucleotide sequence of the plurality is connected to an adjacent polynucleotide sequence of the plurality by one or more cleavage sequence, and wherein the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the polynucleotide sequences of the plurality.

[0695] 2. A multisubunit nucleic acid comprising a plurality of polynucleotide sequences, wherein each polynucleotide sequence of the plurality comprises a self-assembling immunogenic sequence, wherein each polynucleotide sequence of the plurality is connected to an adjacent polynucleotide sequence of the plurality by one or more cleavage sequence, and wherein the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the polynucleotide sequences of the plurality.

[0696] 3. A multisubunit nucleic acid comprising a plurality of polynucleotide sequences, wherein each polynucleotide sequence of the plurality comprises a self-assembling immunogenic sequence further comprising one or more target sequence, wherein each polynucleotide sequence of the plurality is connected to an adjacent polynucleotide sequence of the plurality by one or more cleavage sequence, and wherein the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the polynucleotide sequences of the plurality. A multisubunit nucleic acid encoding a plurality of polypeptides, wherein some or all polypeptides of the plurality comprises either a self-assembling immunogenic peptide, or a self-assembling immunogenic peptide further comprising one or more target peptide, or a combination thereof, wherein each polypeptide of the plurality is connected to an adjacent polypeptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus of one or more of the polypeptides of the plurality. A multisubunit nucleic acid encoding a plurality of polypeptides, wherein each polypeptide comprises a self-assembling immunogenic peptide, wherein each polypeptide of the plurality is connected to an adjacent polypeptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus of one or more of the polypeptides of the plurality. A multisubunit nucleic acid encoding a plurality of polypeptides, wherein each polypeptide comprises a self-assembling immunogenic peptide further comprising one or more target peptide, wherein each polypeptide of the plurality is connected to an adjacent polypeptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus of one or more of the polypeptides of the plurality. The multisubunit nucleic acid according to any one of the preceding paragraphs, wherein total number of the polynucleotide sequences or the polypeptides are not more than 100. The multisubunit nucleic acid according to paragraph 7, wherein total number of the polynucleotide sequences or the polypeptides are between 2-5, 2-10, 10-20, 20- 30, 30-40, 40-50, 50-60, 60-70, 70-80, 80-90, or 90-99. The multisubunit nucleic acid according to any one of the preceding paragraphs, wherein the multisubunit nucleic acid is a DNA or an RNA. The multisubunit nucleic acid according to paragraph 9, wherein the RNA is an mRNA. The multisubunit nucleic acid according to paragraph 10, wherein the mRNA is 0.5 kb to 1 kb, 1 kb to 2 kb, 2 kb to 3 kb, 3 kb to 4 kb, 4 kb to 5 kb, 5 kb to 6 kb, 6 kb to 7 kb, 7 kb to 8 kb, 8 kb to 9 kb, 9 kb to 10 kb, 10 kb to 11 kb, 11 kb to 12 kb, 12 kb to 13 kb, 13 kb to 14 kb, 14 kb to 15 kb, 15 kb to 16 kb, 16 kb to 17 kb, 17 kb to 18 kb, 18 kb to 19 kb, 19 kb to 20 kb in length, or any range therein. The multisubunit nucleic acid according to any one of the paragraphs 10-11 , wherein the mRNA is obtained through a single IVT process or step. The multisubunit nucleic acid according to any one of the paragraphs 1, 3, and 7-

[0697] 12, wherein the target sequence comprises a linker sequence on the 5’ (5 -prime), 3’ (3-prime) or on both sides of the target sequence. The multisubunit nucleic acid according to any one of the paragraphs 1, 3, and 7-

[0698] 13, wherein the linker sequence encodes a linker peptide. The multisubunit nucleic acid according to any one of the paragraphs 4 and 6-14, wherein the target peptide comprises a linker peptide on the C-terminus, N- terminus, or on both ends of the target peptide. The multisubunit nucleic acid according to any one of the paragraphs 14-15, wherein the linker peptide is an amino acid linker, a foldon, a scaffold, or a combination thereof. The multisubunit nucleic acid according to paragraph 16, wherein the linker peptide is the amino acid linker. The multisubunit nucleic acid according to paragraph 17, wherein the amino acid linker comprises 2 to 49 amino acids. The multisubunit nucleic acid according to any one of the paragraphs 17-18, wherein the amino acid linker is a glycine serine linker, a glycine proline linker, a glycine threonine linker, an alanine serine linker, any combination of two amino acids, or a combination thereof. The multisubunit nucleic acid according to paragraph 16, wherein the linker peptide is the foldon. The multisubunit nucleic acid according to paragraph 16, wherein the linker peptide is the scaffold. The multisubunit nucleic acid according to any one of the paragraphs 16-19, wherein the linker peptide comprises the amino acid linker and the foldon. The multisubunit nucleic acid according to any one of the paragraphs 16-19, wherein the linker peptide comprises the amino acid linker and the scaffold. The multisubunit nucleic acid according to any one of the paragraphs 16-19, wherein the linker peptide comprises the amino acid linker, the foldon, and the scaffold. The multisubunit nucleic acid according to paragraph 16, wherein the linker peptide comprises the foldon and the scaffold. The multisubunit nucleic acid according to any one of the paragraphs 4 and 6-25, wherein the linker peptide has an amino acid sequence of any one of SEQ ID NOs: 1-40. The multisubunit nucleic acid according to any one of the paragraphs 1-3 and 7-26, wherein the one or more cleavage sequence encodes one or more cleavage peptides. The multisubunit nucleic acid according to any one of the paragraphs 4-27, wherein the one or more cleavage peptides are optionally connected to each other by a linker peptide. The multisubunit nucleic acid according to paragraph 28, wherein the cleavage peptide is a golgi specific cleavage peptide, a self cleaving peptide, or a combination thereof. The multisubunit nucleic acid according to any one of the paragraphs 4-29, wherein the cleavage peptide has an amino acid sequence of any one of SEQ ID NOs: 41- 55. The multisubunit nucleic acid according to any one of the paragraphs 1-3 and 7-30, wherein the one or more signal sequence encodes one or more signal peptides. The multisubunit nucleic acid according to paragraph 31 , wherein the one or more signal peptides are optionally connected to each other by a linker peptide. The multisubunit nucleic acid according to any one of the paragraphs 4-32, wherein the signal peptide is present on the amino-terminus of one or more of the polypeptides of the plurality. The multisubunit nucleic acid according to any one of the paragraphs 4-33, wherein the signal peptide has an amino acid sequence of any one of SEQ ID NOs: 56-75. The multisubunit nucleic acid according to any one of the paragraphs 1-3 and 7-34, wherein the self-assembling immunogenic sequence encodes a self-assembling immunogenic peptide obtained or derived from LI protein of a papillomavirus. The multisubunit nucleic acid according to paragraph 35, wherein the LI protein comprises a mutation at: a) a codon encoding an amino acid C at the position corresponding to the position 175 of SEQ ID NO: 712, wherein the mutation is a substitution replacing the amino acid C with an amino acid A, b) a codon encoding an amino acid C at the position corresponding to the position 428 of SEQ ID NO: 712, wherein the mutation is a substitution replacing the amino acid C with an amino acid A, or c) a combination thereof. The multisubunit nucleic acid according to any one of the paragraphs 1, 3, and 7- 36, wherein the target sequence encodes a target peptide obtained or derived from L2 protein, El protein, E2 protein, E4 protein, E5 protein, E6 protein, E7 protein, B cell epitope, T cell epitope, or a combination thereof of a papillomavirus. The multisubunit nucleic acid according to any one of the paragraphs 4, and 6-37, wherein the target peptide is inserted into the BC loop, DE loop, EF loop, FG loop, HI loop, or a combination thereof of the LI protein of a papillomavirus. The multisubunit nucleic acid according to paragraph 38, wherein the target peptide is inserted into the DE loop, FG loop, or a combination thereof of the LI protein of a papillomavirus. The multisubunit nucleic acid according to any one of the paragraphs 35-39, wherein the papillomavirus is selected from the group comprising bovine papillomavirus (BPV), canine oral papillomavirus (COPV), cotton tail rabbit papillomavirus (CRPV), european elk papillomavirus (EEPV), rhesus monkey papillomavirus (RhPV), human papillomavirus (HPV), or a combination thereof. The multisubunit nucleic acid according to paragraph 40, wherein the human papillomavirus (HPV) is selected from the group comprising HPV1, HPV2, HPV3, HPV4, HPV5, HPV6, HPV7, HPV8, HPV9, HPV10, HPV11, HPV12, HPV13, HPV14, HPV15, HPV16, HPV17, HPV18, HPV19, HPV20, HPV21, HPV22, HPV23, HPV24, HPV25, HPV26, HPV27, HPV28, HPV29, HPV30, HPV31, HPV32, HPV33, HPV34, HPV35, HPV36, HPV37, HPV38, HPV39, HPV40, HPV41, HPV42, HPV43, HPV44, HPV45, HPV47, HPV48, HPV49, HPV50, HPV51, HPV52, HPV53, HPV54, HPV56, HPV57, HPV58, HPV59, HPV60, HPV61, HPV62, HPV63, HPV65, HPV66, HPV67, HPV68, HPV69, HPV70, HPV71, HPV72, HPV73, HPV74, HPV75, HPV76, HPV77, HPV78, HPV80, HPV81, HPV82, HPV83, HPV84, HPV85, HPV86, HPV87, HPV88, HPV89, HPV90, HPV91, HPV92, HPV93, HPV94, HPV95, HPV96, HPV97, HPV98, HPV99, HPV100, HPV101, HPV102, HPV103, HPV104, HPV105, HPV106, HPV107, HPV108, HPV109, HPV110, HPV111, HPV112, HPV113, HPV114, HPV115, HPV116, HPV117, HPV118, HPV119, HPV120, HPV121, HPV122, HPV123, HPV124, HPV125, HPV126, HPV127, HPV128, HPV129, HPV130, HPV131, HPV132, HPV133, HPV134, HPV135, HPV136, HPV137, HPV138, HPV139, HPV140, HPV141, HPV142, HPV143, HPV144, HPV145, HPV146, HPV147, HPV148, HPV148, HPV150, HPV151, HPV152, HPV153, HPV154, HPV155, HPV156, HPV157, HPV158, HPV159, HPV160, HPV161, HPV162, HPV163, HPV164, HPV165, HPV166, HPV167, HPV168, HPV169, HPV170, HPV171, HPV172, HPV173, HPV174, HPV175, HPV176, HPV177, HPV178, HPV179, HPV180, HPV181, HPV182, HPV183, HPV184, HPV185, HPV186, HPV187, HPV188, HPV189, HPV190, HPV191, HPV192, HPV193, HPV194, HPV195, HPV196, HPV197, HPV198, HPV199, HPV200, HPV201, HPV202, HPV203, HPV204, HPV205, HPV206, HPV207, HPV208, HPV209, HPV210, HPV211, HPV212, HPV213, HPV214, HPV215, HPV216, HPV217, HPV218, HPV219, HPV220, HPV221, HPV222, or a combination thereof. The multisubunit nucleic acid according to paragraph 41, wherein the human papillomavirus is selected from the group comprising HPV5, HPV6, HPV8, HPV11, HPV16, HPV18, HPV26, HPV30, HPV31, HPV33, HPV34, HPV35, HPV39, HPV42, HPV43, HPV44, HPV45, HPV51, HPV52, HPV53, HPV56, HPV58, HPV59, HPV66, HPV67, HPV68, HPV69, HPV70, HPV73, HPV82, HPV85, HPV97, or a combination thereof. The multisubunit nucleic acid according to any one of the paragraphs 35-42, wherein the LI protein has an amino acid sequence of any one of SEQ ID NOs: 76- 1129 or 9409-9444. The multisubunit nucleic acid according to any one of the paragraphs 37-43, wherein the L2 protein has an amino acid sequence according to any of SEQ ID NOs: 1130-3675. The multisubunit nucleic acid according to any one of the paragraphs 37-43, wherein the El protein has an amino acid sequence according to any of SEQ ID NOs: 3676-4768. The multisubunit nucleic acid according to any one of the paragraphs 37-43, wherein the E2 protein has an amino acid sequence according to any of SEQ ID NOs: 4769-5850. The multisubunit nucleic acid according to any one of the paragraphs 37-43, wherein the E4 protein has an amino acid sequence according to any of SEQ ID NOs: 5851-6415. The multisubunit nucleic acid according to any one of the paragraphs 37-43, wherein the E5 protein has an amino acid sequence according to any of SEQ ID NOs: 6416-6713. The multisubunit nucleic acid according to any one of the paragraphs 37-43, wherein the E6 protein has an amino acid sequence according to any of SEQ ID NOs: 6714-7420. The multisubunit nucleic acid according to any one of the paragraphs 37-43, wherein the E7 protein has an amino acid sequence according to any of SEQ ID NOs: 7421-8044. The multisubunit nucleic acid according to any one of the paragraphs 37-43, wherein the B cell epitope has an amino acid sequence according any of SEQ ID NOs: 8045-8654. The multisubunit nucleic acid according to any one of the paragraphs 37-43, wherein the T cell epitope has an amino acid sequence according to any of SEQ ID NOs: 8655-9408 or 9409-9444. A lipid nanoparticle composition comprising a cationic lipid, a phospholipid, a sterol, a PEG-lipid, and the multisubunit nucleic acid according to any one of the preceding paragraphs. The lipid nanoparticle composition according to paragraph 53, wherein the cationic lipid is present in an amount from 10 mol percent to 70 mol percent. The lipid nanoparticle composition according to paragraph 53, wherein the phospholipid is present in an amount from 2 mol percent to 65 mol percent. The lipid nanoparticle composition according to paragraph 53, wherein the sterol is present in an amount from 20 mol percent to 65 mol percent. The lipid nanoparticle composition according to paragraph 53, wherein the PEG- lipid is present in an amount from 0.2 mol percent to 2.0 mol percent. The lipid nanoparticle composition according to paragraph 53, additionally comprising an ionizable polymer. The lipid nanoparticle composition according to paragraph 58, wherein the ionizable polymer is present in an amount from 1 mol percent to 25 mol percent. The lipid nanoparticle composition according to any one of the paragraphs 53-59, wherein the cationic lipid is represented by any one of formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), or SM-102, or ALC-0315, or a combination thereof. The lipid nanoparticle composition according to any one of the paragraphs 53-60, wherein the cationic lipid is represented by formula (I). The lipid nanoparticle composition according to any one of the paragraphs 53-60, wherein the cationic lipid is represented by formula (II). The lipid nanoparticle composition according to any one of the paragraphs 53-60, wherein the cationic lipid is represented by formula (III). The lipid nanoparticle composition according to any one of the paragraphs 53-60, wherein the cationic lipid is represented by formula (IV). The lipid nanoparticle composition according to any one of the paragraphs 53-60, wherein the cationic lipid is represented by formula (V). The lipid nanoparticle composition according to any one of the paragraphs 53-60, wherein the cationic lipid is represented by formula (VI). The lipid nanoparticle composition according to any one of the paragraphs 53-60, wherein the cationic lipid is represented by formula (VII). The lipid nanoparticle composition according to any one of the paragraphs 53-60, wherein the cationic lipid is represented by formula (VIII). A multisubunit peptide encoded by the multisubunit nucleic acid according to any one of paragraphs 1-52. A multisubunit peptide comprising a plurality of polypeptides, wherein some or all polypeptides of the plurality comprises either a self-assembling immunogenic peptide, or a self-assembling immunogenic peptide further comprising one or more target peptide, or a combination thereof, wherein each polypeptide of the plurality is connected to an adjacent polypeptide of the plurality by one or more cleavage peptide and wherein the polypeptide further comprises one or more signal peptide on the amino-terminus of one or more of the polypeptides of the plurality. The multisubunit peptide according to paragraph 70, wherein total number of the polypeptides are not more than 100. The multisubunit peptide according to paragraph 71, wherein total number of the polypeptides are between 2-5, 2-10, 10-20, 20-30, 30-40, 40-50, 50-60, 60-70, 70- 80, 80-90, or 90-99. The multisubunit peptide according to any one of the paragraphs 70-72, wherein the target peptide comprises a linker peptide on the C-terminus, N-terminus, or on both sides of the target peptide. The multisubunit peptide according to paragraph 73, wherein the linker peptide is an amino acid linker, a foldon, a scaffold, or a combination thereof. The multisubunit peptide according to paragraph 74, wherein the linker peptide is the amino acid linker. The multisubunit peptide according to paragraph 75, wherein the amino acid linker comprises 2 to 49 amino acids. The multisubunit peptide according to any one of the paragraphs 75-76, wherein the amino acid linker is a glycine serine linker, a glycine proline linker, a glycine threonine linker, an alanine serine linker, any combination of two amino acids, or a combination thereof. The multisubunit peptide according to paragraph 74, wherein the linker peptide is the foldon. The multisubunit peptide according to paragraph 74, wherein the linker peptide is the scaffold. The multisubunit peptide according to any one of the paragraphs 74-77, wherein the linker peptide comprises the amino acid linker and the foldon. The multisubunit peptide according to any one of the paragraphs 74-77, wherein the linker peptide comprises the amino acid linker and the scaffold. The multisubunit peptide according to any one of the paragraphs 74-77, wherein the linker peptide comprises the amino acid linker, the foldon, and the scaffold. The multisubunit peptide according to paragraph 74, wherein the linker peptide comprises the foldon and the scaffold. The multisubunit peptide according to any one of the paragraphs 73-83, wherein the linker peptide has an amino acid sequence of any one of SEQ ID NOs: 1-40. The multisubunit peptide according to any one of the paragraphs 70-84, wherein the one or more cleavage peptides are optionally connected to each other by a linker peptide. The multisubunit peptide according to paragraph 85, wherein the cleavage peptide is a golgi specific cleavage peptide, a self-cleaving peptide, or a combination thereof. The multisubunit peptide according to paragraph 86, wherein the cleavage peptide has an amino acid sequence of any one of SEQ ID NOs: 41-55. The multisubunit peptide according to any one of the paragraphs 70-87, wherein the signal peptide is present on the amino-terminus of one or more of the polypeptides of the plurality. The multisubunit peptide according to paragraph 88, wherein a second signal peptide is present on the amino-terminus of one or more of the polypeptides of the plurality. The multisubunit peptide according to any one of the paragraphs 88-89, wherein the signal peptide has an amino acid sequence of any one of SEQ ID NOs: 56-75. The multisubunit peptide according to any one of the paragraphs 70-90, wherein the self-assembling immunogenic peptide is obtained or derived from LI protein of a papillomavirus. The multisubunit peptide according to any one of the paragraphs 91, wherein the LI protein comprises a mutation at: a) the position corresponding to the position 175 of SEQ ID NO: 712, wherein the mutation is a substitution replacing the amino acid C with an amino acid A, b) the position corresponding to the position 428 of SEQ ID NO: 712, wherein the mutation is a substitution replacing the amino acid C with an amino acid A. or c) a combination thereof. The multisubunit peptide according to any one of the paragraphs 70-92, wherein the target peptide is obtained or derived from L2 protein, El protein, E2 protein, E4 protein, E5 protein, E6 protein, E7 protein, B cell epitope, T cell epitope, or a combination thereof of a papillomavirus. The multisubunit peptide according to paragraph 93, wherein the target peptide is inserted into the BC loop, DE loop, EF loop, FG loop, HI loop, or a combination thereof of the LI protein of a papillomavirus. The multisubunit peptide according to paragraph 94, wherein the target peptide is inserted into the DE loop, FG loop, or a combination thereof of the LI protein of a papillomavirus. The multisubunit peptide according to any one of the paragraphs 91-95, wherein the papillomavirus is selected from the group comprising bovine papillomavirus (BPV), canine oral papillomavirus (COPV), cotton tail rabbit papillomavirus (CRPV), european elk papillomavirus (EEPV), rhesus monkey papillomavirus (RhPV), human papillomavirus (HPV), or a combination thereof. The multisubunit peptide according to paragraph 96, wherein the human papillomavirus (HPV) is selected from the group comprising HPV1, HPV2, HPV3, HPV4, HPV5, HPV6, HPV7, HPV8, HPV9, HPV10, HPV11, HPV12, HPV13, HPV14, HPV15, HPV16, HPV17, HPV18, HPV19, HPV20, HPV21, HPV22, HPV23, HPV24, HPV25, HPV26, HPV27, HPV28, HPV29, HPV30, HPV31, HPV32, HPV33, HPV34, HPV35, HPV36, HPV37, HPV38, HPV39, HPV40, HPV41, HPV42, HPV43, HPV44, HPV45, HPV47, HPV48, HPV49, HPV50, HPV51, HPV52, HPV53, HPV54, HPV56, HPV57, HPV58, HPV59, HPV60, HPV61, HPV62, HPV63, HPV65, HPV66, HPV67, HPV68, HPV69, HPV70, HPV71, HPV72, HPV73, HPV74, HPV75, HPV76, HPV77, HPV78, HPV80, HPV81, HPV82, HPV83, HPV84, HPV85, HPV86, HPV87, HPV88, HPV89, HPV90, HPV91, HPV92, HPV93, HPV94, HPV95, HPV96, HPV97, HPV98, HPV99, HPV100, HPV101, HPV102, HPV103, HPV104, HPV105, HPV106, HPV107, HPV108, HPV109, HPV110, HPV111, HPV112, HPV113, HPV114, HPV115, HPV116, HPV117, HPV118, HPV119, HPV120, HPV121, HPV122, HPV123, HPV124, HPV125, HPV126, HPV127, HPV128, HPV129, HPV130, HPV131, HPV132, HPV133, HPV134, HPV135, HPV136, HPV137, HPV138, HPV139, HPV140, HPV141, HPV142, HPV143, HPV144, HPV145, HPV146, HPV147, HPV148, HPV148, HPV150, HPV151, HPV152, HPV153, HPV154, HPV155, HPV156, HPV157, HPV158, HPV159, HPV160, HPV161, HPV162, HPV163, HPV164, HPV165, HPV166, HPV167, HPV168, HPV169, HPV170, HPV171, HPV172, HPV173, HPV174, HPV175, HPV176, HPV177, HPV178, HPV179, HPV180, HPV181, HPV182, HPV183, HPV184, HPV185, HPV186, HPV187, HPV188, HPV189, HPV190, HPV191, HPV192, HPV193, HPV194, HPV195, HPV196, HPV197, HPV198, HPV199, HPV200, HPV201, HPV202, HPV203, HPV204, HPV205, HPV206, HPV207, HPV208, HPV209, HPV210, HPV211, HPV212, HPV213, HPV214, HPV215, HPV216, HPV217, HPV218, HPV219, HPV220, HPV221, HPV222, or a combination thereof. The multisubunit peptide according to paragraph 97, wherein the human papillomavirus is selected from the group consisting of HPV5, HPV6, HPV8, HPV11, HPV16, HPV18, HPV26, HPV30, HPV31, HPV33, HPV34, HPV35, HPV39, HPV42, HPV43, HPV44, HPV45, HPV51, HPV52, HPV53, HPV56, HPV58, HPV59, HPV66, HPV67, HPV68, HPV69, HPV70, HPV73, HPV82, HPV85, HPV97, or a combination thereof. The multisubunit peptide according to any one of the paragraphs 91-98, wherein the LI protein has an amino acid sequence of any one of SEQ ID NOs: 76-1129 or 9409-9444. The multisubunit peptide according to any one of the paragraphs 93-99, wherein the L2 protein has an amino acid sequence according to any of SEQ ID NOs: 1130- 3675. The multisubunit peptide according to any one of the paragraphs 93-99, wherein the El protein has an amino acid sequence according to any of SEQ ID NOs: 3676- 4768. The multisubunit peptide according to any one of the paragraphs 93-99, wherein the E2 protein has an amino acid sequence according to any of SEQ ID NOs: 4769- 5850. The multisubunit peptide according to any one of the paragraphs 93-99, wherein the E4 protein has an amino acid sequence according to any of SEQ ID NOs: 5851- 6415. The multisubunit peptide according to any one of the paragraphs 93-99, wherein the E5 protein has an amino acid sequence according to any of SEQ ID NOs: 6416- 6713. The multisubunit peptide according to any one of the paragraphs 93-99, wherein the E6 protein has an amino acid sequence according to any of SEQ ID NOs: 6714- 7420. The multisubunit peptide according to any one of the paragraphs 93-99, wherein the E7 protein has an amino acid sequence according to any of SEQ ID NOs: 7421- 8044. The multisubunit peptide according to any one of the paragraphs 93-99, wherein the B cell epitope has an amino acid sequence according any of SEQ ID NOs: 8045-8654. The multisubunit peptide according to any one of the paragraphs 93-99, wherein the T cell epitope has an amino acid sequence according to any of SEQ ID NOs: 8655-9408. A polypeptide nanoparticle comprising at least 2 or up to 500 polypeptides according to any one of the paragraphs 70-108. The polypeptide nanoparticle according to paragraph 109, comprising a homologous polypeptide, a heterologous polypeptide, an oligomeric complex, or a combination thereof. The polypeptide nanoparticle according to any one of the paragraphs 109-110, wherein the polypeptide nanoparticle is icosahedral, helical, spherical, rod-like, or a combination thereof. A vaccine comprising the multisubunit nucleic acid according to any one of the paragraphs 1-52. A vaccine comprising the multisubunit peptide according to any one of the paragraphs 70-108 or the polypeptide nanoparticle according to any one of the paragraphs 109-111. A vaccine comprising the lipid nanoparticle composition according to any one of the paragraphs 53-68. A method of treating or preventing a disease, comprising administrating to a subject in need thereof the multisubunit nucleic acid according to any one of the paragraphs 1-52 or the vaccine according to any one of the paragraphs 112-113. A method of treating or preventing a disease, comprising administrating to a subject in need thereof the lipid nanoparticle composition according to any one of the paragraphs 53-68 or the vaccine according to paragraph 114. The method according to any one of the paragraphs 115-116, wherein the disease is cervical cancer, vaginal cancer, vulvar cancer, penile cancer, anal cancer, rectal cancer, head and neck cancer, lung cancer, skin cancer, oropharyngeal cancer, or a skin infection (e.g., a noncancerous skin growth, a wart, or a verrucae). Use of the multisubunit nucleic acid sequence according to any one of the paragraphs 1-52 or the vaccine according to any one of the paragraphs 112-113, in the manufacture of a medicament for the treatment or prevention of a disease in a subject. Use of the lipid nanoparticle composition according to any one of the paragraphs 53-68 or the vaccine according to 114, in the manufacture of a medicament for the treatment or prevention of a disease in a subject. The use according to any one of the paragraphs 118-119, wherein the disease is cervical cancer, vaginal cancer, vulvar cancer, penile cancer, anal cancer, rectal cancer, head and neck cancer, lung cancer, skin cancer, oropharyngeal cancer, or a skin infection (e.g., a noncancerous skin growth, a wart, or a verrucae). The present disclosure is further exemplified by the following non limiting examples. It should be understood that the examples are provided to illustrate the disclosure. From the description and the exemplified embodiments and examples, one skilled in the art can make various modifications or adaptations to the disclosure. Such modifications or adaptations are deemed to be within the scope of the spirit of the disclosure.

[0699] Examples

[0700] Example 1: Synthesis of multisubunit nucleic acid sequence (mRNA)

[0701] The plasmid DNA construction, plasmid DNA isolation & linearization, invitro transcription (IVT), transfection etc can be accomplished by general teachings available to a person skilled in the art or using the reference procedure described in PCT Application No. PCT / IB2023 / 000787 or Indian provisional patent Application No. IN202421064707 with appropriate adaptations or modifications.

[0702] Western blot, ELISA and other techniques may be utilized to confirm the expression of the multisubunit peptide.

[0703] Analytical or Immunogenicity assays

[0704] The testing of multisubunit nucleic acid sequence comprises immunizing animals (typically mice) with multisubunit nucleic acid sequence in appropriate formulation following prime-boost immunization strategy at pre-determined dosage amounts. The serum is collected at appropriate intervals and antibody response against the target peptide is measured by ELISA.

[0705] The efficacy of the multisubunit nucleic acid sequence is evaluated by pseudovirus neutralization assays well known to persons skilled in the art. The method typically involves incubating the pseudovirus in the presence of different concentrations of immunised serum containing the antibody of interest (i.e., antibodies produced against the target peptide) and adding this mixture to the cells and incubating it further to measure luminescence to determine inhibitory or neutralization titre. Example 2 (prophetic): Synthesis of multisubunit nucleic acid sequence (mRNA) - bivalent, trivalent, tetravalent, and pentavalent constructs

[0706] Plasmid DNA construction

[0707] The multisubunit nucleic acid sequence comprising at least two (bivalent), three (trivalent), four (tetravalent), or five (pentavalent) self-assembling immunogenic sequences or polynucleotide sequences is codon optimized for human expression. The multisubunit nucleic acid sequence comprises either: a. a plurality of self- assembling immunogenic sequences (each encoding a selfassembling immunogenic peptide), b. a plurality of self- assembling immunogenic sequences (each encoding a selfassembling immunogenic peptide) further comprising a target sequence (encoding a target peptide), or c. their combination.

[0708] Each of the self-assembling immunogenic sequence is separated by one or more cleavage sequence (encoding a cleavage peptide). This multisubunit nucleic acid sequence also includes a signal sequence (encoding a signal peptide) upstream of the first selfassembling immunogenic sequence, and optionally upstream of second and / or subsequent self-assembling immunogenic sequences.

[0709] Alternatively, the multisubunit nucleic acid comprises a plurality of polynucleotide sequences, wherein each polynucleotide sequence comprises either: a. a self-assembling immunogenic sequence (encoding a self-assembling immunogenic peptide), b. a self-assembling immunogenic sequence (encoding a self-assembling immunogenic peptide) further comprising a target sequence (encoding a target peptide), or c. their combination.

[0710] Each of the polynucleotide sequences is separated by one or more cleavage sequence (encoding a cleavage peptide). The multisubunit nucleic acid also includes a signal sequence (encod...

Claims

What is claimed:

1. A multisubunit nucleic acid comprising either a plurality of self-assembling immunogenic sequences or a plurality of self-assembling immunogenic sequences each further comprising one or more target sequence, or a combination thereof, wherein each self-assembling immunogenic sequence is connected to an adjacent self-assembling immunogenic sequence of the plurality by one or more cleavage sequence, and wherein the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the self-assembling immunogenic sequences of the plurality.

2. A multisubunit nucleic acid comprising a plurality of self-assembling immunogenic sequences, wherein each self-assembling immunogenic sequence is connected to an adjacent self-assembling immunogenic sequence of the plurality by one or more cleavage sequence, and wherein the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the self-assembling immunogenic sequences of the plurality.

3. The multisubunit nucleic acid according to claim 2, wherein some or all of the selfassembling immunogenic sequences of the plurality further comprises one or more target sequence, wherein each self-assembling immunogenic sequence of the plurality is connected to an adjacent self-assembling immunogenic sequence of the plurality by one or more cleavage sequence, and wherein the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the self-assembling immunogenic sequences of the plurality.

4. A multisubunit nucleic acid comprising a plurality of self-assembling immunogenic sequences, wherein each self-assembling immunogenic sequence further comprises one or more target sequence, wherein each self-assembling immunogenic sequence of the plurality is connected to an adjacent self-assembling immunogenic sequence of the plurality by one or more cleavage sequence, and wherein the multisubunit nucleic acid further comprises one or more signal sequence upstream of one or more of the selfassembling immunogenic sequences of the plurality.

5. A multisubunit nucleic acid encoding a multisubunit peptide, wherein the multisubunit peptide comprises either a plurality of self-assembling immunogenic peptides or a plurality of self-assembling immunogenic peptide further comprising one or more target peptide, or a combination thereof, wherein each self-assembling immunogenic peptide is connected to an adjacent self-assembling immunogenic peptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus of one or more of the self-assembling immunogenic peptides of the plurality.

6. A multisubunit nucleic acid encoding a multisubunit peptide, wherein the multisubunit peptide comprises a plurality of self-assembling immunogenic peptides, wherein each self-assembling immunogenic peptide of the plurality is connected to an adjacent self-assembling immunogenic peptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus of one or more of the self- assembling immunogenic peptides of the plurality.

7. The multisubunit nucleic acid according to claim 6, wherein some or all of the selfassembling immunogenic peptides of the plurality further comprises one or more target peptide, wherein each self-assembling immunogenic peptide of the plurality is connected to an adjacent self-assembling immunogenic peptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus of one or more of the self-assembling immunogenic peptides of the plurality.

8. A multisubunit nucleic acid encoding a multisubunit peptide, wherein the multisubunit peptide comprises a plurality of self-assembling immunogenic peptides further comprising one or more target peptide, wherein each self-assembling immunogenic peptide of the plurality is connected to an adjacent self-assembling immunogenic peptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus of one or more of the self-assembling immunogenic peptides of the plurality.

9. The multisubunit nucleic acid according to any one of the preceding claims, wherein total number of the self-assembling immunogenic sequences or the selfassembling immunogenic peptides are not more than 100.

10. The multisubunit nucleic acid according to claim 9, wherein total number of the self-assembling immunogenic sequences or the self-assembling immunogenic peptides are between 2-5, 2-10, 10-20, 20-30, 30-40, 40-50, 50-60, 60-70, 70-80, 80-90, or 90-99.

11. The multisubunit nucleic acid according to any one of the preceding claims, wherein the multisubunit nucleic acid is a DNA or an RNA.

12. The multisubunit nucleic acid according to claim 11, wherein the RNA is an mRNA.

13. The multisubunit nucleic acid according to claim 12, wherein, the mRNA is 1 to 20 kb, 1 to 18 kb, 1 to 16 kb, 1 to 14 kb, 1 to 12 kb, 1 to 10 kb, 1 to 9 kb, 1 to 8 kb, 1 to 7 kb,1 to 6 kb, 1 to 5 kb in length, or any range therein.

14. The multisubunit nucleic acid according to any one of the claims 12-13, wherein the mRNA is obtained through a single IVT process or step.

15. The multisubunit nucleic acid according to any one of the claims 1, 3-4, and 9-14, wherein the target sequence comprises a linker sequence on the 5’(5-prime), 3’ (3-prime) or on both sides of the target sequence.

16. The multisubunit nucleic acid according to claim 15, wherein the linker sequence encodes a linker peptide.

17. The multisubunit nucleic acid according to any one of the claims 5 and 7-8, wherein the target peptide comprises a linker peptide on the C-terminus, N-terminus, or on both ends of the target peptide.

18. The multisubunit nucleic acid according to any one of the claims 16-17, wherein the linker peptide is an amino acid linker, a foldon, a scaffold, or a combination thereof.

19. The multisubunit nucleic acid according to claim 18, wherein the linker peptide is the amino acid linker.

20. The multisubunit nucleic acid according to claim 19, wherein the amino acid linker comprises 2 to 49 amino acids.

21. The multisubunit nucleic acid according to any one of the claims 19-20, wherein the amino acid linker is a glycine serine linker, a glycine proline linker, a glycine threonine linker, an alanine serine linker, any combination of two amino acids, or a combination thereof.

22. The multisubunit nucleic acid according to claim 18, wherein the linker peptide is the foldon.

23. The multisubunit nucleic acid according to claim 18, wherein the linker peptide is the scaffold.

24. The multisubunit nucleic acid according to any one of the claims 18-21, wherein the linker peptide comprises the amino acid linker and the foldon.

25. The multisubunit nucleic acid according to any one of the claims 18-21, wherein the linker peptide comprises the amino acid linker and the scaffold.

26. The multisubunit nucleic acid according to any one of the claims 18-21, wherein the linker peptide comprises the amino acid linker, the foldon, and the scaffold.

27. The multisubunit nucleic acid according to claim 18, wherein the linker peptide comprises the foldon and the scaffold.

28. The multisubunit nucleic acid according to any one of the claims 16-27, wherein the linker peptide has an amino acid sequence of any one of SEQ ID NOs: 1-40.

29. The multisubunit nucleic acid according to any one of the claims 1-4 and 9-28, wherein the one or more cleavage sequence encodes one or more cleavage peptides.

30. The multisubunit nucleic acid according to any one of the claims 5-29, wherein the one or more cleavage peptides are optionally connected to each other by a linker peptide.

31. The multisubunit nucleic acid according to claim 30, wherein the cleavage peptide is a golgi specific cleavage peptide, a self cleaving peptide, or a combination thereof.

32. The multisubunit nucleic acid according to any one of the claims 5-31, wherein the cleavage peptide has an amino acid sequence of any one of SEQ ID NOs: 41-55.

33. The multisubunit nucleic acid according to any one of the claims 1-4 and 9-32, wherein the one or more signal sequence encodes one or more signal peptides.

34. The multisubunit nucleic acid according to any one of the claims 5-33, wherein the one or more signal peptides are optionally connected to each other by a linker peptide.

35. The multisubunit nucleic acid according to any one of the claims 33-34, wherein the signal peptide is present on the amino-terminus of one or more of the self-assembling immunogenic peptides of the plurality.

36. The multisubunit nucleic acid according to any one of the claims 5-35, wherein the signal peptide has an amino acid sequence of any one of SEQ ID NOs: 56-75.

37. The multisubunit nucleic acid according to any one of the claims 1-4, wherein the self-assembling immunogenic sequence encodes a self-assembling immunogenic peptide.

38. The multisubunit nucleic acid according to any one of the claims 5-37, wherein the self-assembling immunogenic peptide is obtained or derived from LI protein of a papillomavirus.

39. The multisubunit nucleic acid according to claim 38, wherein the LI protein comprises a mutation at: a) a codon encoding an amino acid C at the position corresponding to the position 175 of SEQ ID NO:712, wherein the mutation is a substitution replacing the amino acid C with an amino acid A, b) a codon encoding an amino acid C at the position corresponding to the position 428 of SEQ ID NO:712, wherein the mutation is a substitution replacing the amino acid C with an amino acid A, or c) a combination thereof.

40. The multisubunit nucleic acid according to any one of the claims 1 and 3-4, wherein the target sequence encodes a target peptide.

41. The multisubunit nucleic acid according to any one of the claims 5 and 7-40, wherein the target peptide is obtained or derived from L2 protein, El protein, E2 protein, E4 protein, E5 protein, E6 protein, E7 protein, B cell epitope, T cell epitope, or a combination thereof of a papillomavirus.

42. The multisubunit nucleic acid according to claim 41, wherein the target peptide is inserted into the BC loop, DE loop, EF loop, FG loop, HI loop, or a combination thereof of the LI protein of a papillomavirus.

43. The multisubunit nucleic acid according to claim 42, wherein the target peptide is inserted into the DE loop, FG loop, or a combination thereof of the LI protein of a papillomavirus.

44. The multisubunit nucleic acid according to any one of the claims 38-43, wherein the papillomavirus is selected from the group comprising bovine papillomavirus (BPV), canine oral papillomavirus (COPY), cotton tail rabbit papillomavirus (CRPV), europeanelk papillomavirus (EEPV), rhesus monkey papillomavirus (RhPV), human papillomavirus (HPV), or a combination thereof.

45. The multisubunit nucleic acid according to claim 44, wherein the human papillomavirus (HPV) is selected from the group comprising HPV1, HPV2, HPV3, HPV4, HPV5, HPV6, HPV7, HPV8, HPV9, HPV10, HPV11, HPV12, HPV13, HPV14, HPV15, HPV16, HPV17, HPV18, HPV19, HPV20, HPV21, HPV22, HPV23, HPV24, HPV25, HPV26, HPV27, HPV28, HPV29, HPV30, HPV31, HPV32, HPV33, HPV34, HPV35, HPV36, HPV37, HPV38, HPV39, HPV40, HPV41, HPV42, HPV43, HPV44, HPV45, HPV47, HPV48, HPV49, HPV50, HPV51, HPV52, HPV53, HPV54, HPV56, HPV57, HPV58, HPV59, HPV60, HPV61, HPV62, HPV63, HPV65, HPV66, HPV67, HPV68, HPV69, HPV70, HPV71, HPV72, HPV73, HPV74, HPV75, HPV76, HPV77, HPV78, HPV80, HPV81, HPV82, HPV83, HPV84, HPV85, HPV86, HPV87, HPV88, HPV89, HPV90, HPV91, HPV92, HPV93, HPV94, HPV95, HPV96, HPV97, HPV98, HPV99, HPV100, HPV101, HPV102, HPV103, HPV104, HPV105, HPV106, HPV107, HPV108, HPV109, HPV110, HPV111, HPV112, HPV113, HPV114, HPV115, HPV116, HPV117, HPV118, HPV119, HPV120, HPV121, HPV122, HPV123, HPV124, HPV125, HPV126, HPV127, HPV128, HPV129, HPV130, HPV131, HPV132, HPV133, HPV134, HPV135, HPV136, HPV137, HPV138, HPV139, HPV140, HPV141, HPV142, HPV143, HPV144, HPV145, HPV146, HPV147, HPV148, HPV148, HPV150, HPV151, HPV152, HPV153, HPV154, HPV155, HPV156, HPV157, HPV158, HPV159, HPV160, HPV161, HPV162, HPV163, HPV164, HPV165, HPV166, HPV167, HPV168, HPV169, HPV170, HPV171, HPV172, HPV173, HPV174, HPV175, HPV176, HPV177, HPV178, HPV179, HPV180, HPV181, HPV182, HPV183, HPV184, HPV185, HPV186, HPV187, HPV188, HPV189, HPV190, HPV191, HPV192, HPV193, HPV194, HPV195, HPV196, HPV197, HPV198, HPV199, HPV200, HPV201, HPV202, HPV203, HPV204, HPV205, HPV206, HPV207, HPV208, HPV209, HPV210, HPV211, HPV212, HPV213, HPV214, HPV215, HPV216, HPV217, HPV218, HPV219, HPV220, HPV221, HPV222, or a combination thereof.

46. The multisubunit nucleic acid according to claim 45, wherein the human papillomavirus is selected from the group comprising HPV5, HPV6, HPV8, HPV11, HPV16, HPV18, HPV26, HPV30, HPV31, HPV33, HPV34, HPV35, HPV39, HPV42, HPV43, HPV44, HPV45, HPV51, HPV52, HPV53, HPV56, HPV58, HPV59, HPV66,HPV67, HPV68, HPV69, HPV70, HPV73, HPV82, HPV85, HPV97, or a combination thereof.

47. The multisubunit nucleic acid according to any one of the claims 38-46, wherein the LI protein has an amino acid sequence of any one of SEQ ID NOs: 76-1129 or 9409- 9444.

48. The multisubunit nucleic acid according to any one of the claims 41-47, wherein the L2 protein has an amino acid sequence according to any one of SEQ ID NOs: 1130- 3675.

49. The multisubunit nucleic acid according to any one of the claims 41-47, wherein the El protein has an amino acid sequence according to any one of SEQ ID NOs: 3676- 4768.

50. The multisubunit nucleic acid according to any one of the claims 41-47, wherein the E2 protein has an amino acid sequence according to any one of SEQ ID NOs: 4769- 5850.

51. The multisubunit nucleic acid according to any one of the claims 41-47, wherein the E4 protein has an amino acid sequence according to any one of SEQ ID NOs: 5851- 6415.

52. The multisubunit nucleic acid according to any one of the claims 41-47, wherein the E5 protein has an amino acid sequence according to any one of SEQ ID NOs: 6416- 6713.

53. The multisubunit nucleic acid according to any one of the claims 41-47, wherein the E6 protein has an amino acid sequence according to any one of SEQ ID NOs: 6714-54. The multisubunit nucleic acid according to any one of the claims 41-47, wherein the E7 protein has an amino acid sequence according to any one of SEQ ID NOs: 7421- 8044.

55. The multisubunit nucleic acid according to any one of the claims 41-47, wherein the B cell epitope has an amino acid sequence according any of SEQ ID NOs: 8045-8654.

56. The multisubunit nucleic acid according to any one of the claims 41-47, wherein the T cell epitope has an amino acid sequence according to any of SEQ ID NOs: 8655- 9408.

57. A lipid nanoparticle composition comprising a cationic lipid, a phospholipid, a sterol, a PEG-lipid, and the multisubunit nucleic acid according to any one of the preceding claims.

58. The lipid nanoparticle composition according to claim 57, wherein the cationic lipid is present in an amount from 10 mol percent to 70 mol percent.

59. The lipid nanoparticle composition according to claim 57, wherein the phospholipid is present in an amount from 2 mol percent to 65 mol percent.

60. The lipid nanoparticle composition according to claim 57, wherein the sterol is present in an amount from 20 mol percent to 65 mol percent.

61. The lipid nanoparticle composition according to claim 57, wherein the PEG-lipid is present in an amount from 0.2 mol percent to 2.0 mol percent.

62. The lipid nanoparticle composition according to claim 57, additionally comprising an ionizable polymer.

63. The lipid nanoparticle composition according to claim 62, wherein the ionizable polymer is present in an amount from 1 mol percent to 25 mol percent.

64. The lipid nanoparticle composition according to any one of the claims 57-63, wherein the cationic lipid is represented by any one of formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), or SM-102, or ALC-0315, or a combination thereof.

65. The lipid nanoparticle composition according to any one of the claims 57-64, wherein the cationic lipid is represented by formula (I).

66. The lipid nanoparticle composition according to any one of the claims 57-64, wherein the cationic lipid is represented by formula (II).

67. The lipid nanoparticle composition according to any one of the claims 57-64, wherein the cationic lipid is represented by formula (III).

68. The lipid nanoparticle composition according to any one of the claims 57-64, wherein the cationic lipid is represented by formula (IV).

69. The lipid nanoparticle composition according to any one of the claims 57-64, wherein the cationic lipid is represented by formula (V).

70. The lipid nanoparticle composition according to any one of the claims 57-64, wherein the cationic lipid is represented by formula (VI).

71. The lipid nanoparticle composition according to any one of the claims 57-64, wherein the cationic lipid is represented by formula (VII).

72. The lipid nanoparticle composition according to any one of the claims 57-64, wherein the cationic lipid is represented by formula (VIII).

73. A multisubunit peptide encoded by the multisubunit nucleic acid according to any one of the claims 1-56.

74. A multisubunit peptide comprising either a plurality of self-assembling immunogenic peptides or a plurality of self-assembling immunogenic peptides further comprising one or more target peptide, or a combination thereof, wherein each selfassembling immunogenic peptide is connected to an adjacent self-assembling immunogenic peptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus of one or more of the self-assembling immunogenic peptides of the plurality.

75. A multisubunit peptide comprising a plurality of self-assembling immunogenic peptides, wherein each self-assembling immunogenic peptide of the plurality is connected to an adjacent self-assembling immunogenic peptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus of one or more of the self-assembling immunogenic peptides of the plurality.

76. The multisubunit peptide according to claim 75, wherein some or all of the selfassembling immunogenic peptides of the plurality further comprises one or more target peptide, wherein each self-assembling immunogenic peptide of the plurality is connected to an adjacent self-assembling immunogenic peptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus of one or more of the self-assembling immunogenic peptides of the plurality.

77. A multisubunit peptide comprising a plurality of self-assembling immunogenic peptides, wherein the plurality of self-assembling immunogenic peptides further comprises one or more target peptide, wherein each self-assembling immunogenic peptide of the plurality is connected to an adjacent self-assembling immunogenic peptide of the plurality by one or more cleavage peptide, and wherein the multisubunit peptide further comprises one or more signal peptide on the amino-terminus of one or more of the self-assembling immunogenic peptides of the plurality.

78. The multisubunit peptide according to any one of the claims 74-77, wherein total number of the self-assembling immunogenic peptides are not more than 100.

79. The multisubunit peptide according to claim 78, wherein total number of the selfassembling immunogenic peptides are between 2-5, 2-10, 10-20, 20-30, 30-40, 40-50, 50- 60, 60-70, 70-80, 80-90, or 90-99.

80. The multisubunit peptide according to any one of the claims 74 and 76-79, wherein the target peptide comprises a linker peptide on the C-terminus, N-terminus or both sides of the target peptide.

81. The multisubunit peptide according to claim 80, wherein the linker peptide is an amino acid linker, a foldon, a scaffold, or a combination thereof.

82. The multisubunit peptide according to claim 81, wherein the linker peptide is the amino acid linker.

83. The multisubunit peptide according to claim 82, wherein the amino acid linker comprises 2 to 49 amino acids.

84. The multisubunit peptide according to any one of the claims 82-83, wherein the amino acid linker is a glycine serine linker, a glycine proline linker, a glycine threonine linker, an alanine serine linker, any combination of two amino acids, or a combination thereof.

85. The multisubunit peptide according to claim 81, wherein the linker peptide is the foldon.

86. The multisubunit peptide according to claim 81, wherein the linker peptide is the scaffold.

87. The multisubunit peptide according to any one of the claims 81-84, wherein the linker peptide comprises the amino acid linker and the foldon.

88. The multisubunit peptide according to any one of the claims 81-84, wherein the linker peptide comprises the amino acid linker and the scaffold.

89. The multisubunit peptide according to any one of the claims 81-84, wherein the linker peptide comprises the amino acid linker, the foldon, and the scaffold.

90. The multisubunit peptide according to claim 81 , wherein the linker peptide comprises the foldon and the scaffold.

91. The multisubunit peptide according to any one of the claims 80-90, wherein the linker peptide has an amino acid sequence of any one of SEQ ID NOs: 1-40.

92. The multisubunit peptide according to any one of the claims 74-91, wherein the one or more cleavage peptides are optionally connected to each other by a linker peptide.

93. The multisubunit peptide according to claim 92, wherein the cleavage peptide is a golgi specific cleavage peptide, a self-cleaving peptide, or a combination thereof.

94. The multisubunit peptide according to claim 93, wherein the cleavage peptide has an amino acid sequence of any one of SEQ ID NOs: 41-55.

95. The multisubunit peptide according to any one of the claims 74-94, wherein the signal peptide is present on the amino-terminus of one or more of the self-assembling immunogenic peptides of the plurality.

96. The multisubunit peptide according to claim 95, wherein a second signal peptide is present on the amino-terminus of one or more self-assembling immunogenic peptides of the plurality.

97. The multisubunit peptide according to any one of the claims 95-96, wherein the signal peptide has an amino acid sequence of any one of SEQ ID NOs: 56-75.

98. The multisubunit peptide according to any one of the claims 74-97, wherein the self-assembling immunogenic peptide is obtained or derived from LI protein of a papillomavirus.

99. The multisubunit peptide according to claim 98, wherein the LI protein comprises a mutation at: a) the position corresponding to the position 175 of SEQ ID NO:712, wherein the mutation is a substitution replacing the amino acid C with an amino acid A, b) the position corresponding to the position 428 of SEQ ID NO:712, wherein the mutation is a substitution replacing the amino acid C with an amino acid A, or c) a combination thereof.

100. The multisubunit peptide according to any one of the claims 74 and 76-99, wherein the target peptide is obtained or derived from L2 protein, El protein, E2 protein, E4 protein, E5 protein, E6 protein, E7 protein, B cell epitope, T cell epitope, or a combination thereof of a papillomavirus.

101. The multisubunit peptide according to claim 100, wherein the target peptide is inserted into the BC loop, DE loop, EF loop, FG loop, HI loop, or combination thereof of the LI protein of a papillomavirus.

102. The multisubunit peptide according to claim 101, wherein the target peptide is inserted into the DE loop, the FG loop, or combination thereof of the LI protein of a papillomavirus.

103. The multisubunit peptide according to any one of the claims 98-102, wherein the papillomavirus is selected from the group comprising bovine papillomavirus (BPV), canine oral papillomavirus (COPV), cotton tail rabbit papillomavirus (CRPV), european elk papillomavirus (EEPV), rhesus monkey papillomavirus (RhPV), human papillomavirus (HPV), or a combination thereof.

104. The multisubunit peptide according to claim 103, wherein the human papillomavirus (HPV) is selected from the group comprising HPV1, HPV2, HPV3, HPV4,HPV5, HPV6, HPV7, HPV8, HPV9, HPV10, HPV11, HPV12, HPV13, HPV14, HPV15, HPV16, HPV17, HPV18, HPV19, HPV20, HPV21, HPV22, HPV23, HPV24, HPV25, HPV26, HPV27, HPV28, HPV29, HPV3O, HPV31, HPV32, HPV33, HPV34, HPV35, HPV36, HPV37, HPV38, HPV39, HPV40, HPV41, HPV42, HPV43, HPV44, HPV45, HPV47, HPV48, HPV49, HPV5O, HPV51, HPV52, HPV53, HPV54, HPV56, HPV57, HPV58, HPV59, HPV6O, HPV61, HPV62, HPV63, HPV65, HPV66, HPV67, HPV68, HPV69, HPV70, HPV71, HPV72, HPV73, HPV74, HPV75, HPV76, HPV77, HPV78, HPV8O, HPV81, HPV82, HPV83, HPV84, HPV85, HPV86, HPV87, HPV88, HPV89, HPV90, HPV91, HPV92, HPV93, HPV94, HPV95, HPV96, HPV97, HPV98, HPV99, HPV1OO, HPV1O1, HPV102, HPV103, HPV104, HPV105, HPV106, HPV107, HPV108, HPV109, HPV11O, HPV111, HPV112, HPV113, HPV114, HPV115, HPV116, HPV117, HPV118, HPV119, HPV120, HPV121, HPV122, HPV123, HPV124, HPV125, HPV126, HPV127, HPV128, HPV129, HPV13O, HPV131, HPV132, HPV133, HPV134, HPV135, HPV136, HPV137, HPV138, HPV139, HPV140, HPV141, HPV142, HPV143, HPV144, HPV145, HPV146, HPV147, HPV148, HPV148, HPV15O, HPV151, HPV152, HPV153, HPV154, HPV155, HPV156, HPV157, HPV158, HPV159, HPV16O, HPV161, HPV162, HPV163, HPV164, HPV165, HPV166, HPV167, HPV168, HPV169, HPV170, HPV171, HPV172, HPV173, HPV174, HPV175, HPV176, HPV177, HPV178, HPV179, HPV18O, HPV181, HPV182, HPV183, HPV184, HPV185, HPV186, HPV187, HPV188, HPV189, HPV190, HPV191, HPV192, HPV193, HPV194, HPV195, HPV196, HPV197, HPV198, HPV199, HPV200, HPV201, HPV202, HPV203, HPV204, HPV205, HPV206, HPV207, HPV2O8, HPV209, HPV210, HPV211, HPV212, HPV213, HPV214, HPV215, HPV216, HPV217, HPV218, HPV219, HPV220, HPV221, HPV222, or a combination thereof.

105. The multisubunit peptide according to claim 104, wherein the human papillomavirus is selected from the group comprising HPV5, HPV6, HPV8, HPV11, HPV16, HPV18, HPV26, HPV30, HPV31, HPV33, HPV34, HPV35, HPV39, HPV42, HPV43, HPV44, HPV45, HPV51, HPV52, HPV53, HPV56, HPV58, HPV59, HPV66, HPV67, HPV68, HPV69, HPV70, HPV73, HPV82, HPV85, HPV97, or a combination thereof.

106. The multisubunit peptide according to any one of the claims 98-105, wherein the LI protein has an amino acid sequence of any one of SEQ ID NOs: 76-1129 or 9409-9444.

107. The multisubunit peptide according to any one of the claims 100-106, wherein the L2 protein has an amino acid sequence according to any of SEQ ID NOs: 1130-3675.

108. The multisubunit peptide according to any one of the claims 100-106, wherein the El protein has an amino acid sequence according to any of SEQ ID NOs: 3676-4768.

109. The multisubunit peptide according to any one of the claims 100-106, wherein the E2 protein has an amino acid sequence according to any of SEQ ID NOs: 4769-5850.

110. The multisubunit peptide according to any one of the claims 100-106, wherein the E4 protein has an amino acid sequence according to any of SEQ ID NOs: 5851-6415.

111. The multisubunit peptide according to any one of the claims 100-106, wherein the E5 protein has an amino acid sequence according to any of SEQ ID NOs: 6416-6713.

112. The multisubunit peptide according to any one of the claims 100-106, wherein the E6 protein has an amino acid sequence according to any of SEQ ID NOs: 6714-7420.

113. The multisubunit peptide according to any one of the claims 100-106, wherein the E7 protein has an amino acid sequence according to any of SEQ ID NOs: 7421-8044.

114. The multisubunit peptide according to any one of the claims 100-106, wherein the B cell epitope has an amino acid sequence according any of SEQ ID NOs: 8045-8654.

115. The multisubunit peptide according to any one of the claims 100-106, wherein the T cell epitope has an amino acid sequence according to any of SEQ ID NOs: 8655-9408.

116. A peptide nanoparticle comprising at least 2 or up to 500 self-assembling immunogenic peptides according to any one of the claims 74-115.

117. The peptide nanoparticle according to claim 116, comprising a homologous selfassembling immunogenic peptide, a heterologous self-assembling immunogenic peptide, an oligomeric complex, or a combination thereof.

118. The peptide nanoparticle according to any one of the claims 116-117, wherein the peptide nanoparticle is icosahedral, helical, spherical, rod-like, or a combination thereof.

119. A vaccine comprising the multisubunit nucleic acid according to any one of the claims 1-56.

120. A vaccine comprising the lipid nanoparticle composition according to any one of the claims 57-72.

121. A vaccine comprising the peptide nanoparticle according to any one of the claims 116-118.

122. A method of treating or preventing a disease, comprising administering to a subject in need thereof the multisubunit nucleic acid according to any one of the claims 1-56 or the vaccine according to any one of the claims 119 or 121.

123. A method of treating or preventing a disease, comprising administering to a subject in need thereof the lipid nanoparticle composition according to any one of the claims 57- 72 or the vaccine according to claim 120.

124. The method according to any one of the claims 122-123, wherein the disease is cervical cancer, vaginal cancer, vulvar cancer, penile cancer, anal cancer, rectal cancer, head and neck cancer, lung cancer, skin cancer, oropharyngeal cancer, or a skin infection (e.g., a noncancerous skin growth, a wart, or a verrucae).

125. Use of the multisubunit nucleic acid according to any one of the claims 1-56 or the vaccine according to any one of the claims 119 or 121, in the manufacture of a medicament for the treatment or prevention of a disease in a subject.

126. Use of the lipid nanoparticle composition according to any one of the claims 57-72 or the vaccine according to claim 120, in the manufacture of a medicament for the treatment or prevention of a disease in a subject.

127. The use according to any one of the claims 125-126, wherein the disease is cervical cancer, vaginal cancer, vulvar cancer, penile cancer, anal cancer, rectal cancer, head and neck cancer, lung cancer, skin cancer, oropharyngeal cancer, or a skin infection (e.g., a noncancerous skin growth, a wart, or a verrucae).

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