Novel RSV RNA molecules and compositions for vaccination

Abstract

To provide a novel artificial RNA coding for antigenic peptides or proteins of RSV and compositions / vaccines comprising the RNA for use as a vaccine for prophylaxis or treatment of RSV infections, particularly in infants, newborns, pregnant women, elderly, and immunocompromised patients.SOLUTION: An artificial RNA comprises: a) at least one heterologous 5' untranslated region (5'-UTR) and / or at least one heterologous 3' untranslated region (3'-UTR); and b) at least one coding sequence operably linked to the 3'-UTR and / or 5'-UTR, encoding at least one antigenic peptide or protein derived from a RSV fusion (F) protein or a fragment or variant thereof.SELECTED DRAWING: None

Description

[Technical Field]

[0001] Preface The present invention relates to artificial RNA suitable for use in treating or preventing respiratory syncytial virus (RSV) infection or disorders associated with such infection. In particular, the artificial RNA of the present invention comprises at least one heterologous untranslated region (UTR), preferably a 3'-UTR and / or a 5'-UTR, and a coding region encoding at least one antigenic peptide or protein derived from RSV, particularly at least one antigenic peptide or protein derived from the RSV fusion (F) protein. The artificial RNA is preferably characterized by increased expression efficiency of the coding region operably linked to the UTR element. The present invention also relates to compositions and vaccines comprising the artificial RNA together with a polymeric carrier, a polycationic protein or peptide, or a lipid nanoparticle (LNP). Furthermore, the present invention relates to kits, particularly kits of parts, comprising the artificial RNA, composition, or vaccine. The present invention further relates to methods for treating or preventing disorders or diseases, and primary and secondary medical uses of the artificial RNA, composition, or vaccine. [Background technology]

[0002] Respiratory syncytial virus (RSV) is an enveloped, non-segmented, negative-strand RNA virus in the genus Pneumovirus, family Paramyxoviridae. It is the most common cause of bronchiolitis and pneumonia in children during the first year of life. RSV also causes repeated infections, including severe lower respiratory tract disease, which can occur in people of all ages, especially the elderly, or those with compromised heart, lung, or immune systems. Passive immunization is currently used to prevent severe illness caused by RSV infection, especially in premature infants, infants with bronchopulmonary dysplasia, or congenital heart disease.

[0003] Recommended treatment for RSV bronchiolitis consists primarily of respiratory support and hydration. No specific antiviral therapy is recommended. The neutralizing monoclonal antibody palivizumab is used for prophylaxis in infants and young children, who are at highest risk for severe infection, but it is very expensive and impractical for widespread use. Currently, there is no licensed / approved RSV vaccine, and the development of a safe and effective RSV vaccine is a global public health priority.

[0004] In vaccine trials in the 1960s, infants and children were immunized with a formalin-inactivated whole-virion RSV preparation (FIRSV) or an equivalent paramyxovirus preparation (FIPIV). After immunization with FI-PIV, 5% of subjects who acquired natural RSV infection during the following RSV season were hospitalized, whereas after immunization with FI-RSV, 80% of subjects who acquired RSV infection were hospitalized, and two children died. The rise in RSV infection following vaccination is a particular challenge for vaccine development against RSV infection.

[0005] Therefore, respiratory syncytial virus (RSV) infection represents the largest unmet childhood vaccine need in developed countries and a significant unmet childhood vaccine need globally. Despite more than 40 years of effort, no RSV vaccine has yet been licensed for use in humans.

[0006] Despite the humanized monoclonal antibody palivizumab, live attenuated vaccine viruses that induce strong immune responses have been developed, but are not recommended for use in certain target groups (infants, children, the elderly, and immunocompromised patients). Furthermore, DNA vectors expressing RSV F proteins with B cell epitopes have been used to induce the production of neutralizing antibodies. In this context, Patent Documents 1 and 2 disclose vectors containing DNA sequences encoding RSV F proteins for use as vaccines. However, the use of DNA as a vaccine can be dangerous due to unwanted insertion into the genome, which can lead to functional gene inhibition, cancer, or the formation of anti-DNA antibodies.

[0007] Patent Document 3 discloses an antigenic composition comprising an RNA sequence encoding an RSV antigenic peptide and a protein selected from fusion protein F, glycoprotein G, short hydrophobic protein SH, matrix protein M, nucleoprotein N, large polymerase L, M2-1 protein, M2-2 protein, phosphoprotein P, nonstructural protein NS1, or nonstructural protein NS2, and a protamine-complexed RNA suitable for intradermal administration.

[0008] Patent Document 4 discloses a vaccine comprising RNA encoding an RSV antigenic peptide and a protein selected from glycoprotein F and glycoprotein G, the RNA being in a lipid nanoparticle.

[0009] Apart from the several approaches cited above, there remains an unmet medical need for an efficient vaccine for the prevention or treatment of RSV infection. [Prior art documents] [Patent documents]

[0010] [Patent Document 1] International Publication No. 2008 / 077527 [Patent Document 2] International Publication No. 96 / 040945 [Patent Document 3] International Publication No. 2015 / 024668 [Patent Document 4] International Publication No. 2017 / 070622 Summary of the Invention [Problem to be solved by the invention]

[0011] Therefore, it is an underlying problem of the invention to provide novel artificial RNAs encoding antigenic peptides or proteins of RSV, and compositions / vaccines comprising said RNAs for use as vaccines for the prevention or treatment of RSV infections, particularly in infants, newborns, pregnant women, the elderly, and immunocompromised patients.

[0012] Furthermore, it is desirable that an RNA-based composition or vaccine have some of the following advantageous characteristics: - Improved translation of RNA constructs at the injection site (e.g., muscle) - Highly efficient induction of RSV antigen-specific immune responses against the encoded antigenic peptides or proteins at very low dose regimens - Suitable for maternal immunization - Suitability for vaccination of infants and / or newborns - Suitable for intramuscular administration - Induction of RSV-specific functional humoral immune responses - Induction of RSV-specific B cell memory - Faster onset of immune defense against RSV - Longevity of the induced immune response to RSV - Induction of broad cellular T cell responses against RSV - Induction of a (local and transient) pro-inflammatory environment - No induction of systemic cytokine or chemokine responses after vaccine administration - Well tolerated, no side effects, non-toxic - Vaccination does not enhance RSV infection - Favorable stability properties of the vaccine - Speed, adaptability, ease and scalability of RSV vaccine manufacturing

[0013] The problems outlined above may be solved by the claimed subject matter.

[0014] definition For clarity and readability, the following definitions are provided. The technical features mentioned in these definitions can be read in each embodiment of the present invention. In the context of these embodiments, further definitions and explanations may be specifically provided.

[0015] Percentages in numerical contexts should be understood as relative to the total number of each item. Otherwise, and unless otherwise specified, percentages should be understood as percentages by weight (wt.-%).

[0016] Adaptive immune response: The term "adaptive immune response" as used herein is recognized and understood by those skilled in the art and is intended to refer, for example, to an antigen-specific response of the immune system (adaptive immune system). Antigen specificity allows for the generation of a response tailored to a particular pathogen or pathogen-infected cells. The ability to produce these tailored responses is typically maintained in the body by "memory cells" (B cells). In the context of the present invention, the antigen is provided by an artificial RNA coding sequence encoding at least one antigenic peptide or protein.

[0017] antigen: The term "antigen" as used herein is understood and understood by those skilled in the art and is intended to mean, for example, a substance that can be recognized by the immune system, preferably the adaptive immune system, and can induce an antigen-specific immune response, for example, by the generation of antibodies and / or antigen-specific T cells as part of the adaptive immune response. Typically, an antigen can be or include a peptide or protein that can be presented to T cells by MHC. Also, for example, fragments, variants, and derivatives of peptides or proteins derived from the RSV F protein containing at least one epitope are considered antigens in the context of the present invention. In the context of the present invention, an antigen can be the translation product of the provided artificial RNA identified herein.

[0018] Antigenic peptides or proteins:The term "antigenic peptide or protein" is recognized and understood by those skilled in the art and is intended to mean, for example, a peptide, protein (or polyprotein) derived from an (antigenic) protein / polyprotein that can stimulate the body's adaptive immune system to provide an adaptive immune response. Thus, an "antigenic peptide or protein" includes at least one epitope (as defined herein) or antigen (as defined herein) of the protein from which it is derived (e.g., in the context of the present invention, a RSV peptide or protein, preferably a RSV F protein or variant thereof).

[0019] Artificial nucleic acid: The term "artificial nucleic acid" as used herein is recognized and understood by those skilled in the art and is intended to mean, for example, an artificial nucleic acid that does not occur in nature. Artificial nucleic acids can be DNA molecules, RNA molecules, or hybrid molecules containing DNA and RNA portions. Typically, artificial nucleic acids can be designed and / or generated by genetic engineering methods to correspond to a desired artificial sequence of nucleotides (heterologous sequence). In this context, an artificial sequence is usually a sequence that cannot occur in nature, i.e., differs from the wild-type sequence by at least one nucleotide. The term "wild-type" as used herein is recognized and understood by those skilled in the art and is intended to mean, for example, a naturally occurring sequence. Furthermore, the term "artificial nucleic acid" is not limited to meaning "one single molecule" but is typically understood to include a collection of essentially identical molecules.

[0020] Artificial RNA:The term "artificial RNA" as used herein is intended to mean RNA that does not occur in nature. In other words, artificial RNA can be understood as a non-natural nucleic acid molecule. Such an RNA molecule can be non-natural due to its individual sequence (which does not occur in nature, e.g., a coding sequence with an altered G / C content, UTR) and / or other modifications, such as structural modifications of non-naturally occurring nucleotides. Typically, artificial RNA can be designed and / or generated by genetic engineering methods to correspond to a desired artificial sequence of nucleotides (heterologous sequence). In this context, an artificial RNA sequence is usually a sequence that does not occur in nature, i.e., differs from the wild-type sequence by at least one nucleotide. The term "artificial RNA" is not limited to meaning "one single molecule" but is typically understood to include a collection of essentially identical molecules. Thus, it can refer to multiple types of essentially identical RNA molecules contained in an aliquot or sample. In the context of the present invention, the RNA of the present invention is an artificial RNA as defined herein.

[0021] Cationic: Unless otherwise clear from the specific context, the term "cationic" means that the respective structure carries a positive charge, either permanently or non-permanently, depending on specific conditions such as pH. Thus, the term "cationic" encompasses both "permanently cationic" and "cationizable."

[0022] Cationizable:The term "cationizable" as used herein means that a compound, group, or atom is positively charged at lower pH levels in the environment in which it is placed, and uncharged at higher pH levels. Also, in non-aqueous environments where the pH value cannot be determined, a cationizable compound, group, or atom is positively charged at high hydrogen ion concentrations and uncharged at low hydrogen ion concentrations or hydrogen ion activities. It depends on the individual properties of the cationizable or polycationizable compound, particularly the pKa of each cationizable group or atom, at which pH or hydrogen ion concentration it is charged or uncharged. In dilute aqueous environments, the proportion of positively charged cationizable compounds, groups, or atoms can be estimated using the so-called Henderson-Hasselbalch equation, which is well known to those skilled in the art. For example, in some embodiments, if a compound or moiety is cationizable, it is preferred that it be positively charged at a pH value of about 1-9, preferably 4-9, 5-8, or even 6-8, more preferably less than 9, less than 8, or less than 7, and most preferably at a physiological pH value, e.g., about 7.3-7.4, i.e., under physiological conditions, particularly the physiological salt conditions of cells in vivo. In other embodiments, the cationizable compound or moiety is primarily neutral at physiological pH values, e.g., about 7.0-7.4, but is preferably positively charged at lower pH values. In some embodiments, the preferred range of pKa for a cationizable compound or moiety is about 5 to about 7.

[0023] Coding sequence / coding region: The terms "coding sequence" or "coding region" and the corresponding abbreviation "cds" as used herein will be recognized and understood by those skilled in the art and are intended to mean a sequence of several nucleotide triplets that can be translated, for example, into a peptide or protein. A coding sequence in the context of the present invention is preferably an RNA sequence consisting of several nucleotides divisible by three, beginning with an initiation codon and preferably ending with a stop codon.

[0024] Composition:In the context of the present invention, a "composition" refers to any type of composition in which a particular component (e.g., an artificial RNA of the present invention associated with an LNP) can be contained together with any additional components, typically at least one pharmaceutically acceptable carrier or excipient. Thus, the composition can be a dry composition such as a powder or granules, or a solid unit such as a lyophilized form or tablet. Alternatively, the composition can be in liquid form, with each component contained independently in a dissolved or dispersed (e.g., suspended or emulsified) form.

[0025] Compound: As used herein, "compound" refers to a chemical substance that is a material made up of molecules with essentially the same chemical structure and properties. In the case of small molecule compounds, the molecules are usually identical in their atomic composition and structural arrangement. In the case of polymers or polymeric compounds, the molecules of the compound are very similar, but not all of them are necessarily identical. For example, a polymer segment described as consisting of 50 monomer units may contain individual molecules with, for example, 48 or 53 monomer units.

[0026] Derived from:The term "derived" as used throughout this specification in reference to nucleic acids, i.e., a nucleic acid "derived from" (another) nucleic acid, means that a nucleic acid derived from (another) nucleic acid shares at least 50%, preferably at least 55%, preferably at least 60%, preferably at least 65%, preferably at least 70%, more preferably at least 75%, more preferably at least 80%, 81%, 82%, 83%, 84%, more preferably at least 85%, 86%, 87%, 88%, 89%, even more preferably at least 90%, 91%, 92%, 93%, 94%, even more preferably at least 95%, 96%, 97%, and particularly preferably at least 98%, 99% sequence identity with the nucleic acid from which it is derived. Those skilled in the art will understand that sequence identity is typically calculated for nucleic acids of the same type, i.e., DNA or RNA sequences. Thus, when DNA is "derived" from RNA, or when RNA is "derived" from DNA, the first step is to convert the RNA sequence to the corresponding DNA sequence (in particular, by substituting uracil (U) with thymidine (T) throughout the sequence), or conversely, to convert the DNA sequence to the corresponding RNA sequence (in particular, by substituting thymidine (T) with uracil (U) throughout the sequence). The sequence identity of the DNA sequence or the sequence identity of the RNA sequence is then determined. Preferably, a nucleic acid "derived" from a nucleic acid also refers to a nucleic acid that has been modified compared to the nucleic acid from which it is derived, for example, to further improve RNA stability and / or to prolong and / or increase protein production. Needless to say, such modifications preferably do not impair the stability of the RNA compared to the nucleic acid from which it is derived.In the context of an amino acid sequence (e.g., an antigenic peptide or protein), the term "derived from" means that the amino acid sequence from which it is derived (from another) amino acid sequence (e.g., RSV F protein) shares at least 50%, preferably at least 55%, preferably at least 60%, preferably at least 65%, preferably at least 70%, more preferably at least 75%, more preferably at least 80%, 81%, 82%, 83%, 84%, more preferably at least 85%, 86%, 87%, 88%, 89%, even more preferably at least 90%, 91%, 92%, 93%, 94%, even more preferably at least 95%, 96%, 97%, and particularly preferably at least 98%, 99% sequence identity with the amino acid sequence from which it is derived.

[0027] Thus, when an antigenic peptide or protein is "derived" from the RSV fusion (F) protein, the antigenic peptide or protein "derived" from the RSV F protein is understood to represent a variant or fragment of the RSV F protein, such as F0 (full-length precursor), F-del, F0_DSCav1, F_DSCav1_mut1, F_DSCav1_mut2, F_DSCav1_mut3, F-del_DSCav1, F-del_DSCav1_mut1, F-del_DSCav1_mut2, F-del_DSCav1_mut3 ​​(described herein). Furthermore, antigenic peptides or proteins "derived" from the RSV F protein (e.g., F0, F-del, F0_DSCav1, F_DSCav1_mut1, F_DSCav1_mut2, F_DSCav1_mut3, F-del_DSCav1, F-del_DSCav1_mut1, F-del_DSCav1_mut2, F-del_DSCav1_mut3) may differ in amino acid sequence as defined above and share a certain percentage of identity. Further preferred examples of RSV F proteins from which antigenic peptides or proteins may be "derived" are shown in Table 1.

[0028] Epitope:The term "epitope" (also referred to in the art as "antigenic determinant") as used herein is recognized and understood by those skilled in the art and is intended to refer to, for example, T cell epitopes and B cell epitopes. T cell epitopes or portions of antigenic peptides or proteins may include fragments, preferably about 6 to about 20 or even more amino acids in length, such as fragments processed and presented by MHC class I molecules, preferably about 8 to about 10, e.g., 8, 9, or 10 amino acids (or even 11 or 12 amino acids), or fragments processed and presented by MHC class II molecules, preferably about 13 to about 20 or even more amino acids in length (these fragments may be selected from any portion of the amino acid sequence). These fragments are typically recognized by T cells in the form of a complex consisting of the peptide fragment and the MHC molecule. That is, the fragments are typically not recognized in their native form. B-cell epitopes are typically fragments located on the outer surface of (native) protein or peptide antigens, preferably having 5 to 15 amino acids, more preferably 5 to 12 amino acids, and even more preferably 6 to 9 amino acids, that can be recognized by antibodies (i.e., in their native form). Such epitopes of proteins or peptides can also be selected from any of the variants of such proteins or peptides mentioned herein. In this context, an antigenic determinant can be a conformational or discontinuous epitope composed of segments of a protein or peptide defined herein that are discontinuous in the amino acid sequence of the protein or peptide defined herein but form the same part in the three-dimensional structure, or a continuous or linear epitope composed of a single polypeptide chain. In the context of the present invention, an epitope can be a translation product of the artificial RNA provided, as specified herein.

[0029] piece:The term "fragment" as used throughout the specification in the context of a nucleic acid or amino acid sequence may typically refer to, for example, a shorter portion of the full-length sequence of the nucleic acid or amino acid sequence. Thus, a fragment typically consists of a sequence identical to a corresponding stretch within the full-length sequence. Preferred fragments of a sequence in the context of the present invention consist of a contiguous stretch of entities, such as nucleotides or amino acids, corresponding to a contiguous stretch of entities in the molecule from which the fragment is derived, and represent at least 5%, 10%, 20%, preferably at least 30%, more preferably at least 40%, more preferably at least 50%, even more preferably at least 60%, even more preferably at least 70%, and most preferably at least 80% of the entire (i.e., full-length) molecule (e.g., RSV F protein) from which the fragment is derived. The term "fragment" as used throughout the specification in reference to a protein or peptide generally refers to a protein or peptide sequence as defined herein, which generally refers to an amino acid sequence (or its encoding nucleic acid molecule) that is N-terminally and / or C-terminally truncated compared to the amino acid sequence of the original (native) protein (or its encoding nucleic acid molecule). Thus, such truncation can occur at either the amino acid level or the corresponding nucleic acid level. Thus, sequence identity with respect to such fragments as defined herein preferably refers to the entire protein or peptide as defined herein, or the entire nucleic acid molecule (encoding) such protein or peptide. In the context of an antigen, such fragments can be about 6 to about 20 or more amino acids in length. For example, fragments processed and presented by MHC class I molecules are preferably about 8 to about 10 amino acids in length, e.g., 8, 9, or 10 amino acids (or 6, 7, 11, or 12 amino acids), or fragments processed and presented by MHC class II molecules are preferably about 13 or more amino acids in length, e.g., 13, 14, 15, 16, 17, 18, 19, 20, or more amino acids in length. These fragments can be selected from any portion of the amino acid sequence.These fragments are usually recognized by T cells in the form of a complex consisting of the peptide fragment and an MHC molecule; i.e., the fragments are usually not recognized in their native form. Fragments of proteins or peptides (e.g., in the context of antigens) may comprise at least one epitope of those proteins or peptides. Furthermore, domains of proteins, such as extracellular, intracellular, or transmembrane domains, and shortened or truncated versions of proteins, may be understood to include fragments of proteins.

[0030] Heterogeneous: The term "heterologous" or "heterologous sequence," as used throughout this specification in the context of a nucleic acid sequence or an amino acid sequence, will be recognized and understood by those of skill in the art to mean a sequence (e.g., DNA, RNA, amino acid) and is intended to mean a sequence derived from another gene, another allele, or another species. Two sequences are generally understood to be "heterologous" when they cannot be derived from the same gene or the same allele. That is, heterologous sequences may be derived from the same organism, but they do not naturally (in nature) occur within the same nucleic acid molecule (e.g., the same RNA) or the same protein.

[0031] Humoral immune response: The terms "humoral immunity" or "humoral immune response" are recognized and understood by those skilled in the art and are intended to refer, for example, to B cell-mediated antibody production and optionally the accompanying processes associated with antibody production. Humoral immune responses can typically be characterized by, for example, Th2 activation and cytokine production, germinal center formation and isotype switching, affinity maturation and memory cell generation, etc. Humoral immunity can also typically refer to antibody effector functions, including neutralization of pathogens and toxins, classical complement activation, and opsonization to facilitate phagocytosis and pathogen elimination.

[0032] (Sequence) Identity:The term "identity" as used throughout this specification in the context of nucleic acid or amino acid sequences is recognized and understood by those skilled in the art and is intended to mean, for example, the percentage of identity between two sequences. To determine the percentage identity between two sequences, e.g., nucleic acid or amino acid sequences defined herein, preferably amino acid sequences encoded by artificial nucleic acid sequences defined herein or the amino acid sequences themselves, the sequences can be aligned for subsequent comparison with each other. Thus, for example, a position in a first sequence can be compared with the corresponding position in a second sequence. If a position in the first sequence is occupied by the same component (residue) as in a position in the second sequence, the two sequences are identical at this position. Otherwise, the sequences differ at this position. If an insertion occurs in the second sequence compared to the first sequence, a gap can be inserted into the first sequence and further alignment can be performed. If a deletion occurs in the second sequence compared to the first sequence, a gap can be inserted into the second sequence and further alignment can be performed. The percentage of identity between two sequences is a function of the number of identical positions divided by the total number of positions, including the positions that are only occupied in one sequence.The percentage of identity between two sequences can be determined using mathematical algorithms.A preferred but non-limiting example of the mathematical algorithm that can be used is the algorithm integrated into BLAST program.This program can identify the sequences that are to some extent identical to the sequences of the present invention.

[0033] Immunogen, immunogenicity: The term "immunogen" or "immunogenic" is understood and understood by those skilled in the art and is intended to mean, for example, a compound capable of stimulating / inducing an immune response. Preferably, the immunogen is a peptide, polypeptide, or protein. An immunogen in the sense of the present invention is a translation product of a provided artificial nucleic acid, preferably RNA, that contains at least one coding sequence encoding at least one antigenic peptide, protein, derived from RSV as defined herein. Typically, the immunogen induces an adaptive immune response.

[0034] Immune response: The term "immune response" is recognized and understood by those skilled in the art and is intended to mean, for example, a specific reaction of the adaptive immune system to a particular antigen (a so-called specific or adaptive immune response), a non-specific reaction of the innate immune system (a so-called non-specific or innate immune response), or a combination thereof.

[0035] Immune system: The term "immune system" is recognized and understood by those skilled in the art and is intended to mean, for example, a system of an organism that can protect the organism from infection. When a pathogen successfully penetrates the organism's physical barriers and invades the organism, the innate immune system provides an immediate but nonspecific response. Even if a pathogen evades this innate response, vertebrates have a second layer of protection, the adaptive immune system. Here, the immune system adapts its response during infection to improve pathogen recognition. This improved response is then retained in the form of immunological memory even after the pathogen is eliminated, allowing the adaptive immune system to launch a faster and more powerful attack each time the pathogen is encountered. Accordingly, the immune system includes the innate immune system and the adaptive immune system. Each of these two parts usually includes a humoral component and a cellular component.

[0036] Innate immune system:The term "innate immune system" (also known as the non-specific or unspecific immune system) is recognized and understood by those skilled in the art and is intended to mean a system that includes cells and mechanisms that, for example, typically provide non-specific protection to a host from infection by other organisms. This means that the cells of the innate system can broadly recognize and respond to pathogens, but, unlike the adaptive immune system, do not confer long-lasting protective immunity to the host. The innate immune system can be activated by, for example, ligands of Toll-like receptors (TLRs), or other adjuvants such as lipopolysaccharide, TNF-α, CD40 ligand, or cytokines, monokines, lymphokines, interleukins, or chemokines, IL-1 through IL-33, IFN-alpha, IFN-beta, IFN-gamma, GM-CSF, G-CSF, M-CSF, LT-beta, TNF-alpha, growth factors, and hGH, ligands of human Toll-like receptors (e.g., TLR1 through TLR10), ligands of murine Toll-like receptors (e.g., TLR1 through TLR13), ligands of NOD-like receptors, ligands of RIG-I-like receptors, immunostimulatory nucleic acids, immunostimulatory RNA (isRNA), CpG-DNA, antibacterial agents, or antiviral agents.

[0037] Lipidoid compounds: Lipidoid compounds (also simply called lipidoids) are lipid-like compounds, i.e. amphiphilic compounds with lipid-like physical properties. In the context of the present invention, the term lipid is considered to encompass lipidoid compounds.

[0038] Monovalent vaccines, monovalent compositions:The terms "monovalent vaccine," "monovalent composition," "univalent vaccine," or "univalent composition" are recognized and understood by those skilled in the art and are intended to refer to, for example, a composition or vaccine containing only one antigen derived from a virus. Thus, the vaccine or composition contains only one RNA species encoding a single antigen against a single organism. The term "monovalent vaccine" includes immunization against a single valency. In the context of the present invention, a monovalent RSV vaccine or composition contains an artificial RNA encoding a single antigenic peptide or protein derived from one specific RSV (e.g., RSV F).

[0039] Nucleic acid: The term "nucleic acid" or "nucleic acid molecule" is recognized and understood by those skilled in the art and is intended to mean, for example, a molecule comprising, preferably consisting of, a nucleic acid component. The term nucleic acid molecule preferably refers to a DNA or RNA molecule. It is preferably used synonymously with the term polynucleotide. Preferably, a nucleic acid or nucleic acid molecule is a polymer comprising or consisting of nucleotide monomers, covalently linked to each other by sugar / phosphate backbone phosphodiester bonds. The term "nucleic acid molecule" also encompasses modified nucleic acid molecules, such as base-, sugar-, or backbone-modified DNA or RNA molecules, as defined herein.

[0040] Nucleic acid sequence / RNA sequence / amino acid sequence: The terms "nucleic acid sequence," "RNA sequence," or "amino acid sequence" are recognized and understood by those of skill in the art and are intended to mean, for example, a specific and particular order of consecutive nucleotides or amino acids, respectively.

[0041] Permanently cationic:The term "permanently cationic" as used herein is recognized and understood by those skilled in the art and means, for example, that the respective compound, group, or atom is positively charged at any pH value or hydrogen ion activity of the environment in which it is placed. Typically, the positive charge is due to the presence of a quaternary nitrogen atom. When a compound has multiple such positive charges, it may be referred to as permanently polycationic, a subcategory of permanently cationic.

[0042] Pharmaceutically effective amount: The terms "pharmaceutically effective amount" or "effective amount" are recognized and understood by those of skill in the art and are intended to mean an amount of a compound (e.g., an artificial RNA of the present invention) that is sufficient to induce, for example, a pharmaceutical effect, in the context of the present invention, an immune response (e.g., against an antigenic peptide, protein, polyprotein defined herein), etc.

[0043] Polyvalent / multivalent vaccines, polyvalent / multivalent compositions:The terms "multivalent vaccine," "multivalent composition," "multivalent vaccine," or "multivalent composition" are recognized and understood by those skilled in the art and are intended to refer, for example, to a composition or vaccine containing antigens from more than one strain of a virus, a composition or vaccine containing different antigens from the same virus, or any combination thereof. These terms describe that the vaccine or composition has more than one valency. In the context of the present invention, a multivalent RSV vaccine includes a vaccine containing artificial RNA encoding antigenic peptides or proteins from several different RSV strains, a vaccine containing artificial RNA encoding different antigens from the same RSV strain, or a combination thereof. In a preferred embodiment, the multivalent RSV vaccine or composition contains more than one, preferably two, three, four, or more different artificial RNA species encoding at least one different antigenic peptide or protein of RSV (e.g., RSV F and RSV M or RSV F and RSV N). Methods for producing multivalent mRNA vaccines are disclosed in PCT application PCT / EP2016 / 082487 or published WO 2017 / 1090134A1.

[0044] "Stabilized Nucleic Acid Molecule" or "Stabilized RNA": The term "stabilized nucleic acid molecule" or "stabilized RNA" refers to a nucleic acid molecule, preferably an RNA molecule, that has been modified to be more stable than an unmodified nucleic acid molecule against decay or degradation due to environmental factors or enzymatic digestion (e.g., exo- or endonuclease degradation). In the context of the present invention, preferably, the stabilized nucleic acid molecule, e.g., stabilized RNA, is stabilized in a cell, such as a prokaryotic or eukaryotic cell, preferably a mammalian cell, such as a human cell. The stabilizing effect may also be exerted outside the cell, e.g., in a buffer solution, e.g., during the manufacturing process of a pharmaceutical composition containing the stabilized nucleic acid molecule.

[0045] T cell response:The terms "cellular immunity" or "cellular immune response" or "cellular T cell response" as used herein are recognized and understood by those skilled in the art and are intended to refer to, for example, the activation of macrophages, natural killer cells (NK), and antigen-specific cytotoxic T lymphocytes, and the release of various cytokines, in response to an antigen. More generally, cellular immunity is based on the activation of cells of the immune system, rather than antibodies. Typically, a cellular immune response can be characterized by the activation of antigen-specific cytotoxic T lymphocytes, which can induce apoptosis of cells, such as specific immune cells, such as dendritic cells or other cells, that present epitopes of foreign antigens on their surface. In the context of the present invention, the antigen is provided by artificial RNA encoding at least one antigenic peptide or protein derived from RSV, preferably inducing a T cell response. The artificial RNA, compositions, and vaccines of the present invention are advantageous in eliciting a cellular T cell response against the RSV F antigen.

[0046] Variant (of arrays): The term "variant," as used throughout the specification in the context of a nucleic acid sequence, is recognized and understood by those of skill in the art and is intended to refer, for example, to a variant of the nucleic acid sequence underlying the nucleic acid sequence. For example, a variant nucleic acid sequence may involve one or more nucleotide deletions, insertions, additions, and / or substitutions compared to the nucleic acid sequence from which the variant is derived. Preferably, a variant of a nucleic acid sequence is at least 40%, preferably at least 50%, more preferably at least 60%, more preferably at least 70%, even more preferably at least 80%, even more preferably at least 90%, and most preferably at least 95% identical to the nucleic acid sequence from which the variant is derived. Preferably, the variant is a functional variant. A "variant" of a nucleic acid sequence may have at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% nucleotide identity over a stretch of 10, 20, 30, 50, 75, or 100 nucleotides of such nucleic acid sequence.

[0047] The term "variant" as used throughout this specification in reference to a protein or peptide is recognized and understood by those skilled in the art and is intended to mean a protein or peptide variant having an amino acid sequence that differs from the original sequence in one or more mutations, such as, for example, one or more substituted, inserted, and / or deleted amino acids. Preferably, these fragments and / or variants have the same biological function or specific activity, such as, for example, their specific antigenic properties, compared to the full-length native protein. "Variant" of a protein or peptide as defined in the context of the present invention may contain conservative amino acid substitutions compared to their native sequences, i.e., the unmutated physiological sequences. These amino acid sequences and the nucleotide sequences encoding them particularly fall under the term "variant" as defined herein. Substitutions in which amino acids from the same class are exchanged for one another are called conservative substitutions. In particular, these are amino acids with aliphatic side chains, positively or negatively charged side chains, aromatic groups in the side chains or amino acids whose side chains can enter into hydrogen bonds, e.g., side chains with hydroxyl functions. This means, for example, that an amino acid having a polar side chain is substituted with another amino acid having a similarly polar side chain, or, for example, that an amino acid characterized by a hydrophobic side chain is substituted with another amino acid having a similarly hydrophobic side chain (e.g., replacing serine (threonine) with threonine (serine) or replacing leucine (isoleucine) with isoleucine (leucine)). Insertions and substitutions can be made, particularly at sequence positions that do not cause changes in the three-dimensional structure or affect binding regions. Changes to the three-dimensional structure due to insertions or deletions can be readily determined, for example, using CD spectroscopy (circular dichroism spectroscopy). A "variant" of a protein or peptide can have at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% amino acid identity over a stretch of 10, 20, 30, 50, 75, or 100 amino acids of such protein or peptide. Preferably, a variant of a protein includes a functional variant of a protein, meaning that the variant exerts the same effect or functionality as the protein from which it is derived.

[0048] 3'-untranslated region, 3'-UTR element, 3'-UTR: The term "3'-untranslated region" or "3'-UTR element" is recognized and understood by those skilled in the art and is intended to mean, for example, a portion of a nucleic acid molecule that is located 3' (i.e., "downstream") of a coding sequence and is typically not translated into protein. Typically, a 3'-UTR is a portion of an mRNA that is located between the coding sequence (cds) of the mRNA and the poly(A) sequence. In the context of the present invention, the term 3'-UTR may also include elements such as poly(A) sequences that are not encoded in the DNA template from which the artificial RNA is transcribed, but are added post-transcriptionally during maturation.

[0049] 5'-untranslated region, 5'-UTR element, 5'-UTR: The term "5'-untranslated region (5'-UTR)" is recognized and understood by those skilled in the art and is intended to refer, for example, to a portion of a nucleic acid molecule located 5' (i.e., "upstream") of a coding sequence and not translated into protein. A 5'-UTR is generally understood to be a specific section of a messenger RNA (mRNA) located 5' of the mRNA's coding sequence. A 5'-UTR typically begins at the transcription start site and ends one nucleotide before the start codon of the coding sequence. Preferably, a 5'-UTR has a length of more than 20, 30, 40, or 50 nucleotides. A 5'-UTR may contain elements for controlling gene expression, also referred to as regulatory elements. Such regulatory elements may be, for example, a ribosome binding site. A 5'-UTR may be post-transcriptionally modified, for example, by the addition of a 5' cap.

[0050] 5'-Terminal Oligopyrimidine Tract (TOP), TOP-UTR:The term "5'-terminal oligopyrimidine tract (TOP)" should be understood as a stretch of pyrimidine nucleotides located in the 5'-terminal region of a nucleic acid molecule, such as the 5'-terminal region of a specific mRNA molecule or the 5'-terminal region of a functional entity (e.g., a transcribed region) of a specific gene. This sequence usually begins with a cytidine corresponding to the transcription start site and is usually followed by a stretch of approximately 3 to 30 pyrimidine nucleotides. For example, a TOP may contain 3 to 30 or more nucleotides. The stretch of pyrimidines, i.e., the 5'-TOP, ends with one nucleotide 5' to the first purine nucleotide downstream of the TOP. Messenger RNAs containing a 5'-terminal oligopyrimidine tract are often referred to as TOP mRNAs. Therefore, genes that provide such messenger RNAs are called TOP genes. The terms "TOP motif" or "5'-TOP motif" should be understood as a nucleic acid sequence corresponding to the 5'-TOP defined above. Therefore, a TOP motif in the context of the present invention is preferably a stretch of pyrimidine nucleotides having a length of 3 to 30 nucleotides. Preferably, a TOP motif consists of at least 3 pyrimidine nucleotides, preferably at least 4 pyrimidine nucleotides, preferably at least 5 pyrimidine nucleotides, more preferably at least 6 nucleotides, more preferably at least 7 nucleotides, and most preferably at least 8 pyrimidine nucleotides, the stretch of pyrimidine nucleotides preferably starting with a cytosine nucleotide at its 5'-end. In TOP genes and TOP mRNAs, the TOP motif preferably starts at its 5'-end with the transcription start site and ends one nucleotide 5' to the first purine residue of the gene or mRNA. A TOP motif in the sense of the present invention is preferably located at the 5'-end of a sequence representing a 5'-UTR or of a sequence encoding a 5'-UTR.Therefore, preferably, a stretch of three or more pyrimidine nucleotides is called a "TOP motif" in the sense of the present invention if this stretch is located at the 5'-end of the respective sequence, such as an artificial nucleic acid, a 5'-UTR element of an artificial nucleic acid, or a nucleic acid sequence derived from the 5'-UTR of a TOP gene described herein. In other words, a stretch of three or more pyrimidine nucleotides that is preferably located not at the 5'-end of the 5'-UTR or 5'-UTR element, but inside the 5'-UTR or 5'-UTR element, is not called a "TOP motif". In some embodiments, the nucleic acid sequence of a 5'-UTR element derived from the 5'-UTR of a TOP gene ends at its 3'-end with a nucleotide that is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 positions upstream of the start codon (e.g., A(U / T)G) of the gene or RNA from which it is derived. Therefore, a 5'-UTR element does not contain any part of a protein-coding sequence. Thus, preferably, the protein-coding portion of at least one nucleic acid sequence, in particular an RNA sequence, is provided solely by a coding sequence.

[0051] Brief description of the invention The present invention is based on the surprising discovery by the inventors that at least one peptide or protein derived from the respiratory syncytial virus (RSV) F protein encoded by the artificial RNA of the present invention can be efficiently expressed in mammalian cells. Even more unexpectedly, the inventors have shown that the artificial RNA of the present invention can induce, for example, specific functional and protective immune responses in cotton rats (see, e.g., Examples 2 and 3). Various optimizations of the RSV F antigen design can further improve the immune response. Furthermore, by selecting a suitable heterologous 5' untranslated region (UTR) and a suitable heterologous 3' untranslated region (UTR), the expression of the RSV F antigen encoded by the artificial nucleic acid RNA can be increased (see, e.g., Example 4). Advantageously, the artificial RNA of the present invention comprising a suitable 3'-UTR / 5'-UTR combination induces a highly efficient antigen-specific immune response against the encoded RSV F. Furthermore, the artificial RNA of the present invention contained in lipid nanoparticles (LNPs) induces a highly efficient antigen-specific immune response against RSV F at a very low dose regimen (see, e.g., Example 3). Furthermore, for example, Examples 8 and 12 provide compositions / vaccines comprising additional artificial RNAs encoding additional antigens, which preferably induce or enhance T cell responses, resulting in a Th1-biased immune response, which is believed to be an important prerequisite for a potential RSV vaccine (Th2-biased responses are associated with enhanced respiratory disease (ERD) in animal models). Furthermore, the compositions are suitable for inducing T cell responses. Therefore, the artificial RNAs and compositions / vaccines comprising the artificial RNAs of the present invention are suitable for inducing an immune response against RSV F in mammalian subjects. Therefore, the artificial RNAs and compositions / vaccines comprising the artificial RNAs are suitable for use as vaccines, for example, as human vaccines, for example, as vaccines for pregnant women or infants.

[0052] In a first aspect, the present invention provides an artificial nucleic acid, preferably an artificial RNA, comprising at least one 5' untranslated region (UTR) and / or at least one 3' untranslated region (UTR), and at least one coding sequence operably linked to the 3'-UTR and / or 5'-UTR, encoding at least one antigenic peptide or protein derived from the RSV F protein, or a fragment or variant thereof.

[0053] In a preferred embodiment, the artificial RNA comprises at least one nucleic acid sequence derived from the 3'-UTR of a gene selected from the ALB7 gene, the alpha-globin gene, PSMB3, CASP1, COX6B1, GNAS, NDUFA1, and RPS9, or a homolog, fragment, or variant of any of these genes.

[0054] In a preferred embodiment, the artificial RNA comprises at least one nucleic acid sequence derived from the 5'-UTR of a gene selected from the RPL32 gene, HSD17B4, ASAH1, ATP5A1, MP68, NDUFA4, NOSIP, RPL31, SLC7A3, TUBB4B, and UBQLN2, or a homolog, fragment, or variant of any of these genes.

[0055] Preferably, the artificial RNA of the present invention is a-1 (HSD17B4 / PSMB3), a-2 (Ndufa4 / PSMB3), a-3 (SLC7A3 / PSMB3), a-4 (NOSIP / PSMB3), a-5 (MP68 / PSMB3), b-1 (UBQLN2 / RPS9), b-2 (ASAH1 / RPS9), b-3 (HSD17B4 / RPS9), b-4 (HSD17B4 / CASP1), b-5 (NOSIP / COX6B1), c-1 (NDUFA4 / RPS 9), c-2(NOSIP / NDUFA1), c-3(NDUFA4 / COX6B1), c-4(NDUFA4 / NDUFA1), c-5(ATP5A1 / PSMB3), d-1(Rpl31 / PSMB3), d-2(ATP5A 1 / CASP1), d-3(SLC7A3 / GNAS), d-4(HSD17B4 / NDUFA1), d-5(Slc7a3 / Ndufa1), e-1(TUBB4B / RPS9), e-2(RPL31 / RPS9), e-3(M P68 / RPS9), e-4(NOSIP / RPS9), e-5(ATP5A1 / RPS9), e-6(ATP5A1 / COX6B1), f-1(ATP5A1 / GNAS), f-2(ATP5A1 / NDUFA1), f-3(H SD17B4 / COX6B1), f-4(HSD17B4 / GNAS), f-5(MP68 / COX6B1), g-1(MP68 / NDUFA1), g-2(NDUFA4 / CASP1), g-3(NDUFA4 / GNAS), g RSV operably linked to a 3'-UTR and a 5'-UTR selected from -4 (NOSIP / CASP1), g-5 (RPL31 / CASP1), h-1 (RPL31 / COX6B1), h-2 (RPL31 / GNAS), h-3 (RPL31 / NDUFA1), h-4 (Slc7a3 / CASP1), h-5 (SLC7A3 / COX6B1), i-1 (SLC7A3 / RPS9), i-2 (RPL32 / ALB7), or i-3 (α-globin gene).It comprises at least one coding sequence encoding at least one antigenic peptide or protein derived from the F protein, with a-1 (HSD17B4 / PSMB3), a-4 (NDUFA4 / PSMB3), c-1 (NDUFA4 / RPS9), e-4 (NOSIP / RPS9), g-2 (NDUFA4 / CASP1), i-2 (RPL32 / ALB7), or i-3 (alpha-globin) being particularly preferred.

[0056] The at least one antigenic peptide or protein derived from the RSV F protein can be a full-length F protein (referred to as "F0", aa 1-574) or a C-terminally deleted F protein ("F-del", aa 1-553), or a fragment or variant thereof.

[0057] At least one antigenic peptide or protein may further comprise mutations that stabilize the antigen or a fragment or variant or functional variant thereof (referred to as "DSCav1", e.g., "F0_DSCav1" or "F-del_DSCav1") in the pre-conformation state / pre-fusion conformation, preferably DSCav1 mutations (S155C, S290C, S190F, and V207L).

[0058] At least one antigenic peptide or protein can be a fusion protein (F(1-103)-GS-F(145-574); F(1-103)-GS-F(145-553)) containing two subunits of mature F, F1 and F2, in one chain, linked via a linker (GS) to enhance protein stability.

[0059] Proteins comprising two subunits of mature F in one chain (referred to as "F2-linker-F1"), e.g., (F(1-103)-GS-F(145-574); F(1-103)-GS-F(145-553), can further comprise a DSCav1 mutation (referred to herein as "mut0").

[0060] A protein comprising two subunits of mature F in one chain (referred to as "F2-linker-F1") can comprise, in addition to the DSCav1 mutation, at least one additional mutation that promotes interprotomer disulfide bonding, such as (S46G, A149C, S215P, Y458C, K465Q; referred to herein as "mut1"), (S46G, E92D, A149C, S215P, Y458C, K465Q; referred to herein as "mut2"), or (S46G, N67I, E92D, A149C, S215P, Y458C, K465Q; referred to herein as "mut3"), (A149C, Y458C, K465Q; referred to herein as "mut4"). 458C; referred to herein as "mut4"), (N183GC, N428C; referred to herein as "mut5"), (Q98C, Q361C, S46G, E92D, L95M, S215P, I217P, I221M, R429K, K465Q; referred to herein as "mut6"), (Q98C, Q361C, L95M, I221M, R429K, referred to herein as "mut7"), or (N183GC, N428C, S46G, N67I, E92D, S215P, K465Q; referred to herein as "mut8"), or a fragment or variant, or functional variant thereof.

[0061] RSV encoded by the artificial RNA of the present invention At least one antigenic peptide or protein derived from the F protein is preferably selected from F0, F-del, F0_DSCav1, F_DSCav1_mut1, F_DSCav1_mut2, F_DSCav1_mut3, F_DSCav1_mut4, F_DSCav1_mut5, F_DSCav1_mut6, F_DSCav1_mut7, F_DSCav1_mut8, F_DSCav1_mut0, F-del_DSCav1, F-del_DSCav1_mut1, F-del_DSCav1_mut2, F-del_DSCav1_mut3, F-del_DSCav1_mut4, F-del_DSCav1_mut5, F-del_DSCav1_mut6, F-del_DSCav1_mut7, F-del_DSCav1_mut8, and F-del_DSCav1_mut0.

[0062] At least one coding sequence may be selected from the group consisting of SEQ ID NOs: 68, 483, 898, 1267, 1636, 2005, 2374, 2743, 3112, 3481, 3850, 4219, 4588, 4957, 5326, 5695, 6064, 6433, 6802, 7171, 7540, 7909, 8279-9683, 11726, 12095, 12464, 12833, 13940, 14309, 14678, 15047, 15416, 15785, 13202, 13571, 13602, 13702, 13802, 13902, 14602, 147 ... 6154, 16523, 16892, 17261, 17630, 17999, 18368, 18737, 19106, 19475, or a fragment or variant of any of these sequences, or an amino acid sequence that is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any of these sequences.

[0063] Preferably, the artificial RNA is selected from the group consisting of SEQ ID NOs: 69 to 77, 484 to 492, 899 to 906, 1268 to 1275, 1637 to 1644, 2006 to 2013, 2375 to 2382, 2744 to 2751, 3113 to 3120, 3482 to 3489, 3851 to 3858, 4220 to 4227, 4589 to 4596, 4958 to 4965, 5327 to 5334, 5696 ~5703, 6065~6072, 6434~6441, 6803~6810, 7172~7179, 7541~7548, 7910~7917, 21363~21384, 11727~11734, 12096~12103, 12465~12472, 12834~12841, 13941~13948, 14310~14317, 14679~14686, 1 5048~15055, 15417~15424, 15786~15793, 13203~13210, 13572~13579, 16155~16162, 16524~16531, 16893~16900, 17262~17269, 17631~17638, 18000~18007, 18369~18376, 18738~18745, 19107~1919 114, 19476-19483, 21389-21410, or a fragment or variant of any of these sequences, or a coding sequence that is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any of these sequences.

[0064] The artificial RNA can comprise a codon-modified coding sequence selected from a C-maximized coding sequence, a CAI-maximized coding sequence, a human codon usage-adapted coding sequence, a G / C content-modified coding sequence, and a G / C-optimized coding sequence, or any combination thereof.

[0065] The artificial RNA can be mRNA, viral RNA, self-replicating RNA, circular RNA, or replicon RNA. In a preferred embodiment, the artificial RNA is mRNA.

[0066] The artificial RNA, preferably mRNA, may further comprise at least one selected from a cap structure, a poly(A) sequence, a poly(C) sequence, a histone stem loop, and / or a 3'-end sequence element.

[0067] The artificial RNA of the present invention preferably comprises or consists of an RNA sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 78 to 482, 11735 to 12094, 21415 to 21417, 21561 to 21563, 21489, 21490, 21635, 21636, or a fragment or variant of any of these (encoding F0), SEQ ID NOs: 493 to 894, 501 to 5016, 5017 to 5018, 5019 to 5020, 5021 to 5022, 5023 to 5024, 5025 to 5026, 5027 to 5028, 5029 to 5030, 5031 to 5032, 5033 to 5034, 5035 to 5036, 5037 to 5038, 5039 to 5040, 5041 to 5042, 5043 to 5044, 5045 to 5046, 5047 to 5048, 5049 to 5050, 5051 to 5051, 5052 to 5053, 5054 to 5054, 5055 to 5055, 5056 to 5057, 5058 to 5059, 5059 to 97, 12104-12463, 21418-21420, 21564-21566, 21491, 21492, 21637, 21638, or any fragment or variant thereof (encoding F-del); SEQ ID NOs: 907-1266, 12473-12832, 21421-21423, 21567-21569, 21493-21495, 21639-21641, or any fragment or variant thereof (encoding F0_DSCav1); SEQ ID NOs: 1276-1635, 8278, 12842-13201, 21424-21641 426, 21570-21572, 21496-21498, 21642-21644 or any fragment or variant thereof (encoding F-del_DSCav1), SEQ ID NOs: 1645-2004, 13949-14308, 21433-21435, 21579-21581, 21505-21507, 21651-21653 or any fragment or variant thereof (encoding F_DSCav1_mut1), SEQ ID NOs: 2014-2373, 14318-14677, 21436-21438, 21582-21584, 21508 to 21510, 21654 to 21656, or any fragment or variant thereof (encoding F-del_DSCav1_mut1), SEQ ID NOs: 2383 to 2742, 14687 to 15046, 21439 to 21441, 21585 to 21587, 21511 to 21513, 21657 to 21659, or any fragment or variant thereof (encoding F_DSCav1_mut2), SEQ ID NOs: 2752 to 3111, 15056 to 15415, 21442 to 21444, 21588 to 21590, 21514 to 21516,21660 to 21662 or any fragment or variant thereof (encoding F-del_DSCav1_mut2), SEQ ID NOs: 3121 to 3480, 15425 to 15784, 21445 to 21447, 21591 to 21593, 21517 to 21519, 21663 to 21665 or any fragment or variant thereof (encoding F_DSCav1_mut3), SEQ ID NOs: 3490 to 3849, 15794 to 16153, 21448 to 21450, 21594 to 21596, 21520 to 21522, 21666 to 21668 or any fragment or variant thereof (encoding F-del_DSCav1_mut3), SEQ ID NOs: 3859 to 4218, 13211 to 13570, 21427 to 21429, 21573 to 21575, 21499 to 21501, 21645 to 21647 or any fragment or variant thereof (encoding F_DSCav1_mut0), SEQ ID NOs: 4228 to 4587, 13580 to 13939, 21430 to 21432, 21576 to 21578, 21502 to 21504, 21648 to 21650 or any fragment or variant thereof (encoding F-del_DSCav1_mut0), SEQ ID NOs: 4597 to 4956, 16163 to 16522, 21451 to 21453, 21597 to 21599, 21523 to 21525, 21669 to 21671 or any fragment or variant thereof (encoding F_DSCav1_mut4), SEQ ID NOs: 4966 to 5325, 16532 to 16891, 21454 to 21456, 21600 to 21602, 21526 to 21528, 21672 to 21674 or any fragment or variant thereof (encoding F-del_DSCav1_mut4), SEQ ID NOs: 5335 to 5694, 16901 to 17260, 21457 to 21459, 21603 to 21605, 21529 to 21531, 21675 to 21677 or any fragment or variant thereof (encoding F_DSCav1_mut5), SEQ ID NOs: 5704 to 6063, 17270 to 17629, 21460 to 21462, 21606 to 21608, 21532 to 21534,21678 to 21680 or any fragment or variant thereof (encoding F-del_DSCav1_mut5), SEQ ID NOs: 6073 to 6432, 17639 to 17998, 21463 to 21465, 21609 to 21611, 21535 to 21537, 21681 to 21683 or any fragment or variant thereof (encoding F_DSCav1_mut6), SEQ ID NOs: 6442 to 6801, 180 SEQ ID NOs: 6811-7170, 18377-18736, 21469-21471, 21615-21617, 21541-21543, 21687-21689, or a fragment or variant of any of these (encoding F-del_DSCav1_mut6), variants (encoding F_DSCav1_mut7), SEQ ID NOs: 7180-7539, 18746-19105, 21472-21474, 21618-21620, 21544-21546, 21690-21692, or fragments or variants thereof (encoding F-del_DSCav1_mut7), SEQ ID NOs: 7549-7908, 19115-19474, 21475-21477, 21621-21622 SEQ ID NOs: 623, 21547-21549, 21693-21695, or any fragment or variant thereof (encoding F_DSCav1_mut8), or SEQ ID NOs: 7918-8277, 19484-19843, 21478-21480, 21624-21626, 21550-21552, 21696-21698, or any fragment or variant thereof (encoding F-del_DSCav1_mut8).

[0068] In a second aspect, the present invention provides a composition comprising the artificial RNA of the first aspect.

[0069] In a preferred embodiment, the composition comprising the artificial RNA of the first aspect comprises at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein from RSV selected from matrix protein M, nucleoprotein N, M2-1 protein, and / or phosphoprotein P, or a combination thereof.

[0070] Matrix protein M, nucleoprotein N, M2-1 protein, M2-2 protein, and / or phosphoprotein P are suitable T cell antigens that promote an efficient T cell response to the composition or vaccine when administered to a subject.

[0071] Further artificial RNAs can include a coding sequence that is identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any of SEQ ID NOs: 9685-9692, 10135-10142, 10638-10645, 11184-11191, 21385-21388, 19845-19852, 20214-20221, 20583-20590, 20952-20959, 21411-21414, or any fragment or variant of any of these sequences.

[0072] Preferably, the further artificial RNA is selected from the group consisting of SEQ ID NOs: 9693 to 10052, 10143 to 10502, 10646 to 11005, 11192 to 11551, 19853 to 20212, 20222 to 20581, 20591 to 20950, 20960 to 21319, 21481 to 21488, 21627 to 21634, 21553 to 2156 0, 21699 to 21706, or a fragment or variant of any of these sequences.

[0073] Preferably, the composition can include an artificial RNA of the present invention complexed, encapsulated, or associated with one or more lipids to form lipid nanoparticles.

[0074] The composition preferably comprises an artificial RNA of the invention complexed with one or more lipids to form lipid nanoparticles (LNPs), the LNPs comprising: (i) at least one cationic lipid as defined herein, preferably a lipid of formula (III), more preferably lipid III-3; (ii) a neutral lipid as defined herein, preferably 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC); (iii) a steroid or steroid analogue as defined herein, preferably cholesterol; and (iv) a PEG-lipid as defined herein, for example, PEG-DMG or PEG-cDMA, preferably a PEGylated lipid of formula (IVa); consists essentially of (i) to (iv) are molar ratios of approximately 20-60% cationic lipid: 5-25% neutral lipid: 25-55% sterol; 0.5-15% PEG-lipid.

[0075] The present invention also relates to an RSV vaccine comprising said artificial RNA or said composition.

[0076] The present invention also relates to the use of artificial RNAs, compositions, and vaccines in the treatment or prevention of infection with RSV.

[0077] In particular, the present invention relates to the use of artificial RNAs, compositions, and vaccines in the treatment or prevention of infection with RSV or disorders associated with such infection.

[0078] The present invention further relates to methods for treating or preventing disorders or diseases in a subject, primary and secondary medical uses of the artificial RNAs, compositions, and vaccines. Additionally, the present invention relates to kits, particularly kits of parts, comprising the artificial RNAs, compositions, and vaccines.

[0079] Detailed Description of the Invention This application is submitted with a Sequence Listing in electronic format (WIPO Standard ST.25), which is part of the description of this application. The information contained in the electronic format of the Sequence Listing submitted with this application is incorporated herein by reference in its entirety. Reference herein to a "SEQ ID NO:" refers to the corresponding nucleic acid or amino acid (aa) sequence in the Sequence Listing with the respective identifier. For many sequences, the Sequence Listing also provides additional details, e.g., regarding specific structural features, sequence optimization, GenBank identifiers, or their coding capabilities. In particular, such information may be included in the WIPO Standard ST.25 Sequence Listing numeric identifiers. <223> Therefore, the numerical identifier <223> The information provided below is expressly incorporated herein in its entirety and should be understood as an integral part of the description of the underlying invention.

[0080] Artificial nucleic acid: In a first aspect, the present invention provides a method for producing a medicament for the treatment of a pulmonary arthritis, comprising: a) at least one heterologous 5' untranslated region (5'-UTR) and / or at least one 3' untranslated region (3'-UTR); b) at least one coding sequence operably linked to the 3'-UTR and / or 5'-UTR, encoding at least one antigenic peptide or protein derived from respiratory syncytial virus (RSV), or a fragment or variant thereof.

[0081] In a preferred embodiment of the first aspect, the present invention provides a method for producing a medicament for the treatment of a pulmonary arthritis, comprising: a) at least one heterologous 5' untranslated region (5'-UTR) and / or at least one heterologous 3' untranslated region (3'-UTR); b) at least one coding sequence operably linked to the 3'-UTR and / or 5'-UTR and encoding at least one antigenic peptide or protein derived from the RSV fusion (F) protein or a fragment or variant thereof.

[0082] Generally, the RNA of the present invention can be composed of a protein coding region and a 5'- and / or 3'-untranslated region (UTR). The 3'-UTR, which is variable in sequence and size, is located between the stop codon and the poly(A) tail. Importantly, the 3'-UTR sequence contains several regulatory motifs that determine RNA turnover, stability, and localization, and is responsible for many aspects of post-transcriptional regulation. In medical applications of RNA (e.g., immunotherapy, vaccination, etc.), regulation of RNA translation into protein is crucial for the safety and efficacy of treatment. The inventors surprisingly discovered that certain combinations of 3'-UTR and / or 5'-UTR act in concert to synergistically enhance the expression of operably linked nucleic acid sequences encoding RSV antigenic peptides or proteins. The artificial RNA molecules having UTR combinations of the present invention are advantageous in that they enable rapid and transient expression of large amounts of RSV antigenic peptides or proteins derived from RSV F. Thus, the artificial RNAs provided herein are particularly useful and suitable for a variety of in vivo applications, including vaccination against RSV.

[0083] Preferably, the artificial RNA can comprise at least one heterologous 5'-UTR and / or at least one heterologous 3'-UTR. In this context, the UTR of the present invention comprises or consists of a nucleic acid sequence derived from the 5'-UTR or 3'-UTR of any naturally occurring gene or a fragment, homolog, or variant thereof. Preferably, the 5'-UTR or 3'-UTR of the present invention is heterologous to at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV F. A suitable heterologous 5'-UTR or heterologous 3'-UTR is derived from a naturally occurring gene (not derived from RSV). In other embodiments, synthetically engineered 5'-UTRs or 3'-UTRs can be used in the context of the present invention.

[0084] In a preferred embodiment, at least one artificial RNA comprises at least one heterologous 3'-UTR.

[0085] Preferably, at least one heterologous 3'-UTR comprises or consists of a nucleic acid sequence derived from the 3'-UTR of a chordate gene, preferably a vertebrate gene, more preferably a mammalian gene, and most preferably a human gene, or from a variant of the 3'-UTR of a chordate gene, preferably a vertebrate gene, more preferably a mammalian gene, and most preferably a human gene.

[0086] Preferably, the artificial RNA of the present invention comprises a 3'-UTR, which may be derived from a gene for an RNA with an extended half-life (providing a stable RNA), such as a 3'-UTR as defined and described below.

[0087] Preferably, at least one heterologous 3'-UTR comprises a nucleic acid sequence derived from the 3'-UTR of a gene that preferably encodes a stable mRNA, or from a homologue, fragment or variant of said gene.

[0088] In a preferred embodiment of the first aspect, the artificial RNA of the present invention comprises at least one heterologous 3'-UTR, wherein said at least one heterologous 3'-UTR comprises a nucleic acid sequence derived from the 3'-UTR of a gene selected from PSMB3, ALB7, alpha-globin (referred to as "muag"), CASP1, COX6B1, GNAS, NDUFA1, and RPS9, or from a homolog, fragment, or variant of any of these genes.

[0089] 3'-UTR from ALB7:In a preferred embodiment, the 3'-UTR comprises or consists of a nucleic acid sequence derived from the 3'-UTR of a vertebrate albumin gene or a variant thereof, preferably derived from the 3'-UTR of a mammalian albumin gene or a variant thereof, more preferably derived from the 3'-UTR of a human albumin gene or a variant thereof, even more preferably derived from the 3'-UTR of the human albumin gene according to GenBank Accession No. NM_000477.5, or a homolog, fragment, or variant thereof. Thus, the artificial RNA of the present invention can comprise a 3'-UTR derived from the ALB7 gene, wherein said 3'-UTR derived from the ALB7 gene comprises or consists of a nucleic acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 35 or 36, or a fragment or variant thereof.

[0090] 3'-UTR from the alpha-globin gene: In a preferred embodiment, the 3'-UTR comprises or consists of a nucleic acid sequence derived from the 3'-UTR of a vertebrate alpha-globin gene (referred to as "muag") or from a variant thereof, preferably derived from the 3'-UTR of a mammalian alpha-globin gene or from a variant thereof, more preferably derived from the 3'-UTR of a human alpha-globin gene or from a variant thereof, even more preferably derived from the 3'-UTR of a human alpha-globin gene. Thus, the RNA of the present invention can comprise a 3'-UTR derived from an alpha-globin gene, wherein said 3'-UTR from an alpha-globin gene comprises or consists of a nucleic acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 37 or 38, or a fragment or variant thereof.

[0091] 3'-UTR from PSMB3:The artificial RNA of the present invention can comprise a 3'-UTR derived from the 3'-UTR of a gene encoding the proteasome subunit beta type 3 (PSMB3) protein, or a homolog, variant, fragment, or derivative thereof. Such a 3'-UTR preferably comprises or consists of a nucleic acid sequence derived from the 3'-UTR of the proteasome subunit beta type 3 (PSMB3) gene, or a homolog, variant, fragment, or derivative thereof, preferably derived from a vertebrate, more preferably a mammalian, and most preferably a human proteasome subunit beta type 3 (PSMB3) gene. The gene preferably encodes the proteasome subunit beta type 3 (PSMB3) protein corresponding to the human proteasome subunit beta type 3 (PSMB3) protein (UniProt Ref. No. P49720, entry version #183 dated August 30, 2017). Thus, the artificial RNA of the present invention may comprise a 3'-UTR derived from the PSMB3 gene, wherein the 3'-UTR derived from the PSMB3 gene comprises or consists of a nucleic acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 23 or 24, or a fragment or variant thereof.

[0092] 3'-UTR from CASP1:The artificial RNA of the present invention can comprise a 3'-UTR derived from the 3'-UTR of a gene encoding the caspase-1 (CASP1) protein, or a homolog, variant, fragment, or derivative thereof. Such a 3'-UTR preferably comprises or consists of the 3'-UTR of the caspase-1 (CASP1) gene, preferably a nucleic acid sequence derived from a vertebrate, more preferably a mammalian, and most preferably a human caspase-1 (CASP1) gene, or a homolog, variant, fragment, or derivative thereof. Thus, the RNA of the present invention can comprise a 3'-UTR derived from the CASP1 gene, wherein the 3'-UTR derived from the CASP1 gene comprises or consists of a nucleic acid sequence identical to SEQ ID NO: 25 or 26 or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical thereto, or a fragment or variant thereof.

[0093] 3'-UTR from COX6B1:The artificial RNA of the present invention can comprise a 3'-UTR derived from the 3'-UTR of the COX6B1 gene encoding the cytochrome c oxidase subunit 6B1 (COX6B1) protein, or a homolog, variant, fragment, or derivative thereof. Such a 3'-UTR preferably comprises or consists of a nucleic acid sequence derived from the 3'-UTR of the cytochrome c oxidase subunit 6B1 (COX6B1) gene, preferably from a vertebrate, more preferably a mammal, and most preferably a human, cytochrome c oxidase subunit 6B1 (COX6B1) gene, or a homolog, variant, fragment, or derivative thereof. The gene preferably encodes a cytochrome c oxidase subunit 6B1 (COX6B1) protein corresponding to the human cytochrome c oxidase subunit 6B1 (COX6B1) protein (UniProt Ref. No. P14854, entry version #166 dated August 30, 2017). Thus, the artificial RNA of the present invention may comprise a 3'-UTR derived from the COX6B1 gene, wherein the 3'-UTR derived from the COX6B1 gene comprises or consists of a nucleic acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 27 or 28, or a fragment or variant thereof.

[0094] 3'-UTR from GNAS:The artificial RNA of the present invention can comprise a 3'-UTR derived from the 3'-UTR of the GNAS gene encoding the guanine nucleotide-binding protein G(s) subunit alpha isoform short (GNAS) protein, or a homolog, variant, fragment, or derivative thereof. Such a 3'-UTR preferably comprises or consists of a nucleic acid sequence derived from the 3'-UTR of the guanine nucleotide-binding protein G(s) subunit alpha isoform short (GNAS) gene, preferably from a vertebrate, more preferably a mammalian, guanine nucleotide-binding protein G(s) subunit alpha isoform short (GNAS) gene, or a homolog, variant, fragment, or derivative thereof. The gene preferably encodes the guanine nucleotide-binding protein G(s) subunit alpha isoform short (GNAS) protein corresponding to the human guanine nucleotide-binding protein G(s) subunit alpha isoform short (GNAS) protein (UniProt Ref. No. P63092, entry version #153 dated August 30, 2017). Thus, the artificial RNA of the present invention may comprise a 3'-UTR derived from the GNAS gene, wherein the 3'-UTR derived from the GNAS gene comprises or consists of a nucleic acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 29 or 30, or a fragment or variant thereof.

[0095] 3'-UTR from NDUFA1:The artificial RNA of the present invention can comprise a 3'-UTR derived from the 3'-UTR of a gene encoding the NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 1 (NDUFA1) protein, or a homolog, variant, fragment, or derivative thereof. Such a 3'-UTR preferably comprises or consists of a nucleic acid sequence derived from the 3'-UTR of the NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 1 (NDUFA1) gene, preferably from a vertebrate, more preferably a mammalian, NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 1 (NDUFA1) gene, or a homolog, variant, fragment, or derivative thereof. The gene can preferably encode the NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 1 (NDUFA1) protein corresponding to the human NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 1 (NDUFA1) protein (UniProt Ref. No. O15239, entry version #152 of August 30, 2017). Thus, the artificial RNA of the present invention can comprise a 3'-UTR derived from the NDUFA1 gene, wherein the 3'-UTR derived from the NDUFA1 gene comprises or consists of a nucleic acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 31 or 32, or a fragment or variant thereof.

[0096] 3'-UTR from RPS9:The artificial RNA of the present invention can comprise a 3'-UTR derived from the 3'-UTR of a gene encoding the 40S ribosomal protein S9 (RPS9) protein, or a homolog, variant, fragment, or derivative thereof. Such a 3'-UTR preferably comprises or consists of the 3'-UTR of the 40S ribosomal protein S9 (RPS9) gene, preferably a nucleic acid sequence derived from a vertebrate, more preferably a mammalian, most preferably a human 40S ribosomal protein S9 (RPS9) gene, or a homolog, variant, fragment, or derivative thereof. The gene preferably encodes a 40S ribosomal protein S9 (RPS9) protein corresponding to the human 40S ribosomal protein S9 (RPS9) protein (UniProt Ref. No. P46781, entry version #179 dated August 30, 2017). Thus, the artificial RNA of the present invention may comprise a 3'-UTR derived from the RPS9 gene, wherein the 3'-UTR derived from the RPS9 gene comprises or consists of a nucleic acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 33 or 34, or a fragment or variant thereof.

[0097] Additional 3'-UTR:In embodiments, the artificial RNAs defined herein comprise a 3'-UTR described in WO 2016 / 107877. In this context, the disclosure of WO 2016 / 107877 regarding 3'-UTR sequences is incorporated herein by reference. Particularly suitable 3'-UTRs are SEQ ID NOS: 1-24 and 49-318 of WO 2016 / 107877, or fragments or variants of these sequences. Thus, the 3'-UTR of the artificial RNAs of the present invention can comprise or consist of the RNA sequence corresponding to the nucleic acid sequence according to SEQ ID NOS: 1-24 and 49-318 of WO 2016 / 107877. In other embodiments, the artificial RNAs defined herein comprise a 3'-UTR described in WO 2017 / 036580. In this context, the disclosure of WO 2017 / 036580 regarding 3'-UTR sequences is incorporated herein by reference. Particularly suitable 3'-UTRs are SEQ ID NOS: 152-204 of WO 2017 / 036580, or fragments or variants of these sequences. Thus, the 3'-UTR of the artificial RNA of the present invention can comprise or consist of an RNA sequence corresponding to the nucleic acid sequence according to SEQ ID NOS: 152-204 of WO 2017 / 036580.

[0098] According to a preferred embodiment, the artificial RNA comprises at least one heterologous 5'-UTR.

[0099] In a preferred embodiment, at least one artificial nucleic acid as defined herein, in particular an RNA as defined herein, may comprise at least one heterologous 5'-UTR.

[0100] Preferably, at least one 5'-UTR comprises or consists of a nucleic acid sequence derived from the 5'-UTR of a chordate gene, preferably a vertebrate gene, more preferably a mammalian gene, and most preferably a human gene, or from a variant of the 5'-UTR of a chordate gene, preferably a vertebrate gene, more preferably a mammalian gene, and most preferably a human gene.

[0101] Preferably, the artificial RNA of the present invention comprises a 5'-UTR, which may be derived from a gene for an RNA with an extended half-life (providing a stable RNA), such as a 5'-UTR as defined and described below.

[0102] Preferably, at least one heterologous 5'-UTR comprises a nucleic acid sequence derived from the 5'-UTR of a gene that preferably encodes a stable mRNA, or from a homologue, fragment or variant of said gene.

[0103] In a preferred embodiment of the first aspect, the artificial RNA of the present invention comprises at least one heterologous 5'-UTR, wherein said at least one heterologous 5'-UTR comprises a nucleic acid sequence derived from the 5'-UTR of a gene selected from HSD17B4, RPL32, ASAH1, ATP5A1, MP68, NDUFA4, NOSIP, RPL31, SLC7A3, TUBB4B, and UBQLN2, or derived from a homolog, fragment, or variant of any of these genes.

[0104] 5'-UTR from RPL32:The artificial RNA of the present invention can comprise a 5'-UTR derived from the 5'-UTR of a gene encoding 60S ribosomal protein L32, or a homolog, variant, fragment, or derivative thereof, wherein the 5'-UTR preferably lacks a 5'-TOP motif. Such a 5'-UTR preferably comprises or consists of a nucleic acid sequence derived from the 5'-UTR of the 60S ribosomal protein L32 (RPL32) gene, preferably from a vertebrate, more preferably a mammal, and most preferably a human 60S ribosomal protein L32 (RPL32) gene, or a homolog, variant, fragment, or derivative thereof, wherein the 5'-UTR preferably does not comprise the 5'-TOP motif of the gene. The gene preferably encodes 60S ribosomal protein L32 (RPL32), corresponding to human 60S ribosomal protein L32 (RPL32) (UniProt Ref. No. P62899, entry version #138 dated August 30, 2017). Thus, the artificial RNA of the present invention may comprise a 5'-UTR derived from the RPL32 gene, wherein the 5'-UTR derived from the RPL32 gene comprises or consists of a nucleic acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 21 or 22, or a fragment or variant thereof.

[0105] 5'-UTR from HSD17B4:The artificial RNA of the present invention may comprise a 5'-UTR derived from the 5'-UTR of a gene encoding 17-beta-hydroxysteroid dehydrogenase 4, or a homolog, variant, fragment, or derivative thereof, and preferably lacks a 5'-TOP motif. Such a 5'-UTR comprises or consists of a nucleic acid sequence derived from the 5'-UTR of the 17-beta-hydroxysteroid dehydrogenase 4 (also known as peroxisomal multifunctional enzyme type 2) gene, preferably from a vertebrate, more preferably from a mammal, and most preferably from a human 17-beta-hydroxysteroid dehydrogenase 4 (HSD17B4) gene, or a homolog, variant, fragment, or derivative thereof, and preferably does not contain the 5'-UTR of the gene. The gene preferably encodes a 17-beta-hydroxysteroid dehydrogenase 4 protein corresponding to human 17-beta-hydroxysteroid dehydrogenase 4 (UniProt Ref. No. Q9BPX1, entry version #139 of August 30, 2017), or a homolog, variant, fragment, or derivative thereof. Thus, the artificial RNA of the present invention can comprise a 5'-UTR derived from the HSD17B4 gene, wherein the 5'-UTR derived from the HSD17B4 gene comprises or consists of a nucleic acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 1 or 2, or a fragment or variant thereof.

[0106] 5'-UTR from ASAH1:The artificial RNA of the present invention can comprise a 5'-UTR derived from the 5'-UTR of a gene encoding acid ceramidase (ASAH1), or a homolog, variant, fragment, or derivative thereof. Such a 5'-UTR preferably comprises or consists of a nucleic acid sequence derived from the 5'-UTR of the acid ceramidase (ASAH1) gene, preferably from a vertebrate, more preferably a mammal, and most preferably a human acid ceramidase (ASAH1) gene, or a homolog, variant, fragment, or derivative thereof. The gene preferably encodes an acid ceramidase (ASAH1) protein corresponding to human acid ceramidase (ASAH1) (UniProt Ref. No. Q13510, entry version #177 dated June 7, 2017), or a homolog, variant, fragment, or derivative thereof. Thus, the artificial RNA of the present invention may comprise a 5'-UTR derived from the ASAH1 gene, wherein the 5'-UTR derived from the ASAH1 gene comprises or consists of a nucleic acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 3 or 4, or a fragment or variant thereof.

[0107] 5'-UTR from ATP5A1:The artificial RNA of the present invention can comprise a 5'-UTR derived from the 5'-UTR of a gene encoding mitochondrial ATP synthase subunit alpha (ATP5A1), or a homolog, variant, fragment, or derivative thereof, wherein the 5'-UTR preferably lacks a 5'-TOP motif. Such a 5'-UTR preferably comprises or consists of the 5'-UTR of the mitochondrial ATP synthase subunit alpha (ATP5A1) gene, preferably a nucleic acid sequence derived from a vertebrate, more preferably a mammalian, most preferably a human mitochondrial ATP synthase subunit alpha (ATP5A1) gene, or a homolog, variant, fragment, or derivative thereof, wherein the 5'-UTR preferably does not comprise the 5'-TOP motif of the gene. The gene can preferably encode a mitochondrial ATP synthase subunit alpha protein corresponding to human acidic mitochondrial ATP synthase subunit alpha (UniProt Ref. No. P25705, entry version #208 of August 30, 2017), or a homolog, variant, fragment, or derivative thereof. Thus, the artificial RNA of the present invention may comprise a 5'-UTR derived from the ATP5A1 gene, wherein the 5'-UTR derived from the ATP5A1 gene comprises or consists of a nucleic acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 5 or 6, or a fragment or variant thereof.

[0108] 5'-UTR from MP68:The artificial RNA of the present invention can comprise a 5'-UTR derived from the 5'-UTR of a gene encoding MP68 or its homolog, fragment, or variant. Such a 5'-UTR preferably comprises or consists of a nucleic acid sequence derived from the 5'-UTR of the 6.8 kDa mitochondrial proteolipid (MP68) gene, preferably from a vertebrate, more preferably a mammalian, 6.8 kDa mitochondrial proteolipid (MP68) gene, or its homolog, variant, fragment, or derivative. The gene preferably encodes the 6.8 kDa mitochondrial proteolipid (MP68) protein corresponding to the human 6.8 kDa mitochondrial proteolipid (MP68) protein (UniProt Ref. No. P56378, entry version #127 dated February 15, 2017). Thus, the artificial RNA of the present invention can comprise a 5'-UTR derived from the MP68 gene, wherein the 5'-UTR derived from the MP68 gene comprises or consists of a nucleic acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 7 or 8, or a fragment or variant thereof.

[0109] 5'-UTR from NDUFA4:The artificial RNA of the present invention can include a 5'-UTR derived from the 5'-UTR of a gene encoding cytochrome c oxidase subunit (NDUFA4), or a homolog, fragment, or variant thereof. Such a 5'-UTR preferably comprises or consists of a nucleic acid sequence derived from the 5'-UTR of a cytochrome c oxidase subunit (NDUFA4) gene, preferably a vertebrate, more preferably a mammalian, cytochrome c oxidase subunit (NDUFA4) gene, or a homolog, variant, fragment, or derivative thereof. The gene preferably encodes a cytochrome c oxidase subunit (NDUFA4) protein corresponding to the human cytochrome c oxidase subunit (NDUFA4) protein (UniProt Ref. No. O00483, entry version #149 dated August 30, 2017). Thus, the artificial RNA of the present invention may comprise a 5'-UTR derived from the NDUFA4 gene, wherein the 5'-UTR derived from the NDUFA4 gene comprises or consists of a nucleic acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 9 or 10, or a fragment or variant thereof.

[0110] 5'-UTR from NOSIP:The artificial RNA of the present invention can comprise a 5'-UTR derived from the 5'-UTR of a gene encoding a nitric oxide synthase-interacting protein (NOSIP) or a homolog, variant, fragment, or derivative thereof. Such a 5'-UTR preferably comprises or consists of a nucleic acid sequence derived from the 5'-UTR of a nitric oxide synthase-interacting protein (NOSIP) gene, preferably a vertebrate, more preferably a mammalian, most preferably a human nitric oxide synthase-interacting protein (NOSIP) gene, or a homolog, variant, fragment, or derivative thereof. The gene can preferably encode a nitric oxide synthase-interacting protein (NOSIP) protein corresponding to the human nitric oxide synthase-interacting protein (NOSIP) protein (UniProt Ref. No. Q9Y314, entry version #130 dated June 7, 2017). Thus, the artificial RNA of the present invention can comprise a 5'-UTR derived from the NOSIP gene, wherein the 5'-UTR derived from the NOSIP gene comprises or consists of a nucleic acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 11 or 12, or a fragment or variant thereof.

[0111] 5'-UTR from RPL31:The artificial RNA of the present invention can comprise a 5'-UTR derived from the 5'-UTR of a gene encoding 60S ribosomal protein L31, or a homolog, variant, fragment, or derivative thereof, wherein the 5'-UTR preferably lacks a 5'-TOP motif. Such a 5'-UTR preferably comprises or consists of a nucleic acid sequence derived from the 5'-UTR of the 60S ribosomal protein L31 (RPL31) gene, preferably a vertebrate, more preferably a mammalian 60S ribosomal protein L31 (RPL31) gene, or a homolog, variant, fragment, or derivative thereof, wherein the 5'-UTR preferably does not include the 5'-TOP motif of the gene. The gene preferably encodes 60S ribosomal protein L31 (RPL31) corresponding to human 60S ribosomal protein L31 (RPL31) (UniProt Ref. No. P62899, entry version #138 dated August 30, 2017). Thus, the artificial RNA of the present invention may comprise a 5'-UTR derived from the RPL31 gene, wherein the 5'-UTR derived from the RPL31 gene comprises or consists of a nucleic acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 13 or 14, or a fragment or variant thereof.

[0112] 5'-UTR from SLC7A3:The artificial RNA of the present invention can comprise a 5'-UTR derived from the 5'-UTR of a gene encoding a cationic amino acid transporter 3 (solute carrier family 7 member 3, SLC7A3) protein, or a homolog, variant, fragment, or derivative thereof. Such a 5'-UTR preferably comprises or consists of a nucleic acid sequence derived from the 5'-UTR of a cationic amino acid transporter 3 (SLC7A3) gene, preferably a vertebrate, more preferably a mammalian cationic amino acid transporter 3 (SLC7A3) gene, or a homolog, variant, fragment, or derivative thereof. The gene preferably encodes a cationic amino acid transporter 3 (SLC7A3) protein corresponding to the human cationic amino acid transporter 3 (SLC7A3) protein (UniProt Ref. No. Q8WY07, entry version #139 dated August 30, 2017). Thus, the artificial RNA of the present invention may comprise a 5'-UTR derived from the SLC7A3 gene, wherein the 5'-UTR derived from the SLC7A3 gene comprises or consists of a nucleic acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 15 or 16, or a fragment or variant thereof.

[0113] 5'-UTR from TUBB4B:The artificial RNA of the present invention may comprise a 5'-UTR derived from the 5'-UTR of a gene encoding the tubulin beta-4B chain (TUBB4B) protein, or a homolog, variant, fragment, or derivative thereof. Such a 5'-UTR preferably comprises or consists of a nucleic acid sequence derived from the 5'-UTR of a tubulin beta-4B chain (TUBB4B) gene, preferably from a vertebrate, more preferably a mammal, and most preferably a human, tubulin beta-4B chain (TUBB4B) gene, or a homolog, variant, fragment, or derivative thereof. The gene preferably encodes a tubulin beta-4B chain (TUBB4B) protein corresponding to the human tubulin beta-4B chain (TUBB4B) protein (UniProt Ref. No. Q8WY07, entry version #142 dated August 30, 2017). Thus, the artificial RNA of the present invention may comprise a 5'-UTR derived from the TUBB4B gene, wherein the 5'-UTR derived from the TUBB4B gene comprises or consists of a nucleic acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 17 or 18, or a fragment or variant thereof.

[0114] 5'-UTR from UBQLN2:The artificial RNA of the present invention can include a 5'-UTR derived from the 5'-UTR of a gene encoding the ubiquilin-2 (UBQLN2) protein, or a homolog, variant, or fragment thereof. Such a 5'-UTR preferably comprises or consists of a nucleic acid sequence derived from the 5'-UTR of the ubiquilin-2 (UBQLN2) gene, preferably from a vertebrate, more preferably a mammalian, ubiquilin-2 (UBQLN2) gene, or a homolog, variant, fragment, or derivative thereof. The gene preferably encodes the ubiquilin-2 (UBQLN2) protein corresponding to UniProt Ref. No. Q9UHD9, entry version #151 of August 30, 2017. Thus, the artificial RNA of the present invention can comprise a 5'-UTR derived from the UBQLN2 gene, wherein the 5'-UTR derived from the UBQLN2 gene comprises or consists of a nucleic acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 19 or 20, or a fragment or variant thereof.

[0115] Additional 5'-UTR:In an embodiment, the artificial RNA defined herein comprises a 5'-UTR described in WO 2013 / 143700. In this context, the disclosure of WO 2013 / 143700 regarding 5'-UTR sequences is incorporated herein by reference. Particularly suitable 5'-UTRs are SEQ ID NOs: 1-1363, 1395, 1421, and 1422 of WO 2013 / 143700, or fragments or variants of these sequences. In this context, the 5'-UTR of the artificial RNA of the present invention preferably comprises or consists of the RNA sequence corresponding to the nucleic acid sequence according to SEQ ID NOs: 1-1363, 1395, 1421, and 1422 of WO 2013 / 143700. In another embodiment, the artificial RNA of the present invention comprises a 5'-UTR described in WO 2016 / 107877. In this regard, the disclosure of WO 2016 / 107877 regarding 5'-UTR sequences is incorporated herein by reference. Particularly preferred 5'-UTRs are the nucleic acid sequences set forth in SEQ ID NOS: 25-30 and 319-382 of WO 2016 / 107877, or fragments or variants of these sequences. In this context, it is particularly preferred that the 5'-UTR of the artificial RNA comprises or consists of the RNA sequence corresponding to the nucleic acid sequence set forth in SEQ ID NOS: 25-30 and 319-382 of WO 2016 / 107877. In other embodiments, the artificial RNA of the present invention comprises a 5'-UTR described in WO 2017 / 036580. In this context, the disclosure of WO 2017 / 036580 regarding 5'-UTR sequences is incorporated herein by reference. Particularly preferred 5'-UTRs are the nucleic acid sequences according to SEQ ID NOs: 1 to 151 of WO 2017 / 036580, or fragments or variants of these sequences. In this context, it is particularly preferred that the 5'-UTR of the artificial RNA comprises or consists of an RNA sequence corresponding to a nucleic acid sequence according to SEQ ID NOs: 1 to 151 of WO 2017 / 036580.

[0116] The inventors have discovered that certain combinations of at least one heterologous 5'-UTR and / or at least one heterologous 3'-UTR advantageously enhance translation of at least one coding sequence operably linked to the 3'-UTR and / or 5'-UTR encoding at least one antigenic peptide or protein derived from the RSV F protein in target tissue (e.g., muscle, dermis).

[0117] Therefore, at least one heterologous 5'-UTR as defined herein and at least one heterologous 3'-UTR as defined herein preferably act synergistically to enhance the production (i.e., translation) of antigenic peptides or proteins from the artificial RNA of the invention. These advantageous combinations of 5'-UTRs and 3'-UTRs are shown below. The abbreviations introduced below, namely "a-1", "a-2", "a-3", "a-4", "a-5", "b-1", "b-2", "b-3", "b-4", "b-5", "c-1", "c-2", "c-3", "c-4", "c-5", "d-1", "d-2", "d-3", "d-4", "d-5", "e-1", "e-2", "e-3", "e-4", "e-5", "e-6", "f-1", "f-2", "f-3", "f-4", "f-5", "g-1", "g-2", "g-3", "g-4", "g-5", "h-1", "h-2", "h-3", "h-4", "h-5", "i-1", "i-2", "i-3", each of which is used throughout the specification of the present invention, represent one advantageous combination of 5'-UTR and / or 3'UTR of the present invention.

[0118] Therefore, in a preferred embodiment of the first aspect, the artificial RNA of the present invention comprises: a-1. at least one 5'-UTR derived from the 5'-UTR of the HSD17B4 gene or its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the PSMB3 gene or its corresponding RNA sequence, homolog, fragment, or variant; or a-2. at least one 5'-UTR derived from the 5'-UTR of the NDUFA4 gene or its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the PSMB3 gene or its corresponding RNA sequence, homolog, fragment, or variant; or a-3. at least one 5'-UTR derived from the 5'-UTR of the SLC7A3 gene or from its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the PSMB3 gene or from its corresponding RNA sequence, homolog, fragment, or variant; or a-4. at least one 5'-UTR derived from the 5'-UTR of the NOSIP gene or its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the PSMB3 gene or its corresponding RNA sequence, homolog, fragment, or variant; or a-5. at least one 5'-UTR derived from the 5'-UTR of the MP68 gene or from its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the PSMB3 gene or from its corresponding RNA sequence, homolog, fragment, or variant; or b-1. at least one 5'-UTR derived from the 5'-UTR of the UBQLN2 gene or its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the RPS9 gene or its corresponding RNA sequence, homolog, fragment, or variant; or b-2. at least one 5'-UTR derived from the 5'-UTR of the ASAH1 gene or its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the RPS9 gene or its corresponding RNA sequence, homolog, fragment, or variant; or b-3. at least one 5'-UTR derived from the 5'-UTR of the HSD17B4 gene or its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the RPS9 gene or its corresponding RNA sequence, homolog, fragment, or variant; or b-4. at least one 5'-UTR derived from the 5'-UTR of the HSD17B4 gene or from its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the CASP1 gene or from its corresponding RNA sequence, homolog, fragment, or variant; or b-5. at least one 5'-UTR derived from the 5'-UTR of the NOSIP gene or its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the COX6B1 gene or its corresponding RNA sequence, homolog, fragment, or variant; or c-1. at least one 5'-UTR derived from the 5'-UTR of the NDUFA4 gene or its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the RPS9 gene or its corresponding RNA sequence, homolog, fragment, or variant; or c-2. at least one 5'-UTR derived from the 5'-UTR of the NOSIP gene or its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the NDUFA1 gene or its corresponding RNA sequence, homolog, fragment, or variant; or c-3. at least one 5'-UTR derived from the 5'-UTR of the NDUFA4 gene or its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the COX6B1 gene or its corresponding RNA sequence, homolog, fragment, or variant; or c-4. at least one 5'-UTR derived from the 5'-UTR of the NDUFA4 gene or from its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the NDUFA1 gene or from its corresponding RNA sequence, homolog, fragment, or variant; or c-5. at least one 5'-UTR derived from the 5'-UTR of the ATP5A1 gene or its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the PSMB3 gene or its corresponding RNA sequence, homolog, fragment, or variant; or d-1. at least one 5'-UTR derived from the 5'-UTR of the RPL31 gene or from its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the PSMB3 gene or from its corresponding RNA sequence, homolog, fragment, or variant; or d-2. at least one 5'-UTR derived from the 5'-UTR of the ATP5A1 gene or its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the CASP1 gene or its corresponding RNA sequence, homolog, fragment, or variant; or d-3. at least one 5'-UTR derived from the 5'-UTR of the SLC7A3 gene or from its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the GNAS gene or from its corresponding RNA sequence, homolog, fragment, or variant; or d-4. at least one 5'-UTR derived from the 5'-UTR of the HSD17B4 gene or its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the NDUFA1 gene or its corresponding RNA sequence, homolog, fragment, or variant; or d-5. at least one 5'-UTR derived from the 5'-UTR of the SLC7A3 gene or from its corresponding RNA sequence, homologue, fragment or variant, and at least one 3'-UTR derived from the 3'-UTR of the NDUFA1 gene or from its corresponding RNA sequence, homologue, fragment or variant; or e-1. at least one 5'-UTR derived from the 5'-UTR of the TUBB4B gene or its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the RPS9 gene or its corresponding RNA sequence, homolog, fragment, or variant; or e-2. at least one 5'-UTR derived from the 5'-UTR of the RPL31 gene or from its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the RPS9 gene or from its corresponding RNA sequence, homolog, fragment, or variant; or e-3. at least one 5'-UTR derived from the 5'-UTR of the MP68 gene or from its corresponding RNA sequence, homologue, fragment or variant, and at least one 3'-UTR derived from the 3'-UTR of the RPS9 gene or from its corresponding RNA sequence, homologue, fragment or variant; or e-4. at least one 5'-UTR derived from the 5'-UTR of the NOSIP gene or its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the RPS9 gene or its corresponding RNA sequence, homolog, fragment, or variant; or e-5. at least one 5'-UTR derived from the 5'-UTR of the ATP5A1 gene or from its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the RPS9 gene or from its corresponding RNA sequence, homolog, fragment, or variant; or e-6. at least one 5'-UTR derived from the 5'-UTR of the ATP5A1 gene or its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the COX6B1 gene or its corresponding RNA sequence, homolog, fragment, or variant; or f-1. at least one 5'-UTR derived from the 5'-UTR of the ATP5A1 gene or its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the GNAS gene or its corresponding RNA sequence, homolog, fragment, or variant; or f-2. at least one 5'-UTR derived from the 5'-UTR of the ATP5A1 gene or its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the NDUFA1 gene or its corresponding RNA sequence, homolog, fragment, or variant; or f-3. at least one 5'-UTR derived from the 5'-UTR of the HSD17B4 gene or its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the COX6B1 gene or its corresponding RNA sequence, homolog, fragment, or variant; or f-4. at least one 5'-UTR derived from the 5'-UTR of the HSD17B4 gene or from its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the GNAS gene or from its corresponding RNA sequence, homolog, fragment, or variant; or f-5. at least one 5'-UTR derived from the 5'-UTR of the MP68 gene or from its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the COX6B1 gene or from its corresponding RNA sequence, homolog, fragment, or variant; or g-1. At least one 5'-UTR derived from the 5'-UTR of the MP68 gene or from its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the NDUFA1 gene or from its corresponding RNA sequence, homolog, fragment, or variant; or g-2. at least one 5'-UTR derived from the 5'-UTR of the NDUFA4 gene or its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the CASP1 gene or its corresponding RNA sequence, homolog, fragment, or variant; or g-3. at least one 5'-UTR derived from the 5'-UTR of the NDUFA4 gene or from its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the GNAS gene or from its corresponding RNA sequence, homolog, fragment, or variant; or g-4. at least one 5'-UTR derived from the 5'-UTR of the NOSIP gene or its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the CASP1 gene or its corresponding RNA sequence, homolog, fragment, or variant; or g-5. at least one 5'-UTR derived from the 5'-UTR of the RPL31 gene or from its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the CASP1 gene or from its corresponding RNA sequence, homolog, fragment, or variant; or h-1. at least one 5'-UTR derived from the 5'-UTR of the RPL31 gene or from its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the COX6B1 gene or from its corresponding RNA sequence, homolog, fragment, or variant; or h-2. at least one 5'-UTR derived from the 5'-UTR of the RPL31 gene or from its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the GNAS gene or from its corresponding RNA sequence, homolog, fragment, or variant; or h-3. at least one 5'-UTR derived from the 5'-UTR of the RPL31 gene or from its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the NDUFA1 gene or from its corresponding RNA sequence, homolog, fragment, or variant; or h-4. at least one 5'-UTR derived from the 5'-UTR of the SLC7A3 gene or from its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the CASP1 gene or from its corresponding RNA sequence, homolog, fragment, or variant; or h-5. at least one 5'-UTR derived from the 5'-UTR of the SLC7A3 gene or from its corresponding RNA sequence, homologue, fragment or variant, and at least one 3'-UTR derived from the 3'-UTR of the COX6B1 gene or from its corresponding RNA sequence, homologue, fragment or variant; or i-1. at least one 5'-UTR derived from the 5'-UTR of the SLC7A3 gene or from its corresponding RNA sequence, homolog, fragment, or variant, and at least one 3'-UTR derived from the 3'-UTR of the RPS9 gene or from its corresponding RNA sequence, homolog, fragment, or variant; or i-2. at least one 5'-UTR derived from the 5'-UTR of the RPL32 gene or from its corresponding RNA sequence, homologue, fragment or variant, and at least one 3'-UTR derived from the 3'-UTR of the ALB7 gene or from its corresponding RNA sequence, homologue, fragment or variant; or i-3. At least one 3'-UTR derived from the 3'-UTR of the alpha globin gene or from its corresponding RNA sequence, homologue, fragment or variant.

[0119] Preferably, the artificial RNA of the present invention is a-1 (HSD17B4 / PSMB3), a-2 (Ndufa4 / PSMB3), a-3 (SLC7A3 / PSMB3), a-4 (NOSIP / PSMB3), a-5 (MP68 / PSMB3), b-1 (UBQLN2 / RPS9), b-2 (ASAH1 / RPS9), b-3 (HSD17B4 / RPS9), b-4 (HSD17B4 / CASP1), b-5 (NOSIP / COX6B1), c-1 (NDUFA4 / RPS9) , c-2(NOSIP / NDUFA1), c-3(NDUFA4 / COX6B1), c-4(NDUFA4 / NDUFA1), c-5(ATP5A1 / PSMB3), d-1(Rpl31 / PSMB3), d-2(ATP5A1 / C ASP1), d-3(SLC7A3 / GNAS), d-4(HSD17B4 / NDUFA1), d-5(Slc7a3 / Ndufa1), e-1(TUBB4B / RPS9), e-2(RPL31 / RPS9), e-3(MP68 / R PS9), e-4(NOSIP / RPS9), e-5(ATP5A1 / RPS9), e-6(ATP5A1 / COX6B1), f-1(ATP5A1 / GNAS), f-2(ATP5A1 / NDUFA1), f-3(HSD17B4 / COX6B1), f-4(HSD17B4 / GNAS), f-5(MP68 / COX6B1), g-1(MP68 / NDUFA1), g-2(NDUFA4 / CASP1), g-3(NDUFA4 / GNAS), g-4(NOSIP / CASP1), g-5 (RPL31 / CASP1), h-1 (RPL31 / COX6B1), h-2 (RPL31 / GNAS), h-3 (RPL31 / NDUFA1), h-4 (Slc7a3 / CASP1), h-5 (SLC7A3 / COX6B1), i-1 (SLC7A3 / RPS9), i-2 (RPL32 / ALB7), or i-3 (α-globin gene) operably linked to a 3'-UTR and a 5'-UTR selected from the group consisting of: g-5 (RPL31 / CASP1), h-1 (RPL31 / COX6B1), h-2 (RPL31 / GNAS), h-3 (RPL31 / NDUFA1), h-4 (Slc7a3 / CASP1), h-5 (SLC7A3 / COX6B1), i-1 (SLC7A3 / RPS9), i-2 (RPL32 / ALB7), or i-3 (α-globin gene). At least one coding sequence encoding at least one antigenic peptide or protein derived from the RSV F protein described herein.

[0120] In a particularly preferred embodiment of the first aspect, the artificial RNA of the present invention comprises UTR elements of a-1 (HSD17B4 / PSMB3), a-4 (NDUFA4 / PSMB3), c-1 (NDUFA4 / RPS9), e-4 (NOSIP / RPS9), g-2 (NDUFA4 / CASP1), i-2 (RPL32 / ALB7), or i-3 (alpha-globin (muag)).

[0121] In a particularly preferred embodiment of the first aspect, the artificial RNA of the present invention comprises a UTR element related to a-1 (HSD17B4 / PSMB3).

[0122] The present invention relates to an artificial nucleic acid, preferably an RNA suitable for vaccination, comprising at least one heterologous 5'-UTR as defined above and / or at least one heterologous 3'-UTR as defined above and at least one coding sequence operably linked to said 3'-UTR and / or 5'-UTR, wherein said coding sequence encodes at least one antigenic peptide or protein derived from respiratory syncytial virus ("RSV"), or a fragment or variant thereof.

[0123] As used herein, the term "respiratory syncytial virus" or the corresponding abbreviation "RSV" is not limited to a particular viral strain, variant, serotype, or isolate, including respiratory syncytial virus of any origin.

[0124] According to various embodiments, the artificial RNA, preferably the coding sequence of the artificial RNA, comprises or consists of a nucleic acid sequence derived from a virus, the NCBI classification ID ("NCBI-ID") of which is provided in List 1 below.

[0125] List 1: RSV virus strains: Human orthopneumovirus, HRSV (NCBI-ID 11250); Human respiratory syncytial virus A, HRSV-A, respiratory syncytial virus group A (NCBI-ID 208893); Human respiratory syncytial virus A strain Long, Human respiratory syncytial virus (subgroup A / strain Long) (NCBI-ID 11260); Human respiratory syncytial virus A2, Human respiratory syncytial virus (strain A2), HRSVA (NCBI-ID 11259); Human respiratory syncytial virus (strain RSB1734), (NCBI-ID 11253); Human respiratory syncytial virus (strain RSB5857) (NCBI-ID 11254); Human respiratory syncytial virus (strain RSB6190), (NCBI-ID 11255); Human respiratory syncytial virus (strain RSB6256), (NCBI-ID 11256); Human respiratory syncytial virus (strain RSB642), (NCBI-ID 11252); Human respiratory syncytial virus (strain RSB6614), (NCBI-ID 11257); Human respiratory syncytial virus B, HRSV-B, respiratory syncytial virus group B, (NCBI-ID 208895); Human respiratory syncytial virus 9320 (NCBI-ID 253182); Human respiratory syncytial virus B1 (NCBI-ID 79692); Human respiratory syncytial virus (subgroup B / strain 18537), (NCBI-ID 11251); Human respiratory syncytial virus (subgroup B / strain 8 / 60), (NCBI-ID 11258); Human respiratory syncytial virus S2, (NCBI-ID 410078); Human respiratory syncytial virus strain RSS-2, (NCBI-ID 11261); Unclassified human respiratory syncytial virus, (NCBI-ID 410233); Human respiratory syncytial virus (strain RSP112 / Sweden / 02-03), (NCBI-ID 410237); Human respiratory syncytial virus (strain RSP120 / Sweden / 02-03), (NCBI-ID 410238); Human respiratory syncytial virus (strain RSP121 / Sweden / 02-03), (NCBI-ID 410239); Human respiratory syncytial virus (strain RSP122 / Sweden / 02-03), (NCBI-ID 410247); Human respiratory syncytial virus (strain RSP13 / Sweden / 02-03), (NCBI-ID 410241);Human respiratory syncytial virus (strain RSP140 / Sweden / 02-03), (NCBI-ID 410248); Human respiratory syncytial virus (strain RSP16 / Sweden / 02-03), (NCBI-ID 410242); Human respiratory syncytial virus (strain RSP171 / Sweden / 02-03), (NCBI-ID 410246); Human respiratory syncytial virus (strain RSP183 / Sweden / 02-03), (NCBI-ID 410249); Human respiratory syncytial virus (strain RSP191 / Sweden / 02-03), (NCBI-ID 410240); Human respiratory syncytial virus (strain RSP199 / Sweden / 02-03), (NCBI-ID 410250); Human respiratory syncytial virus (strain RSP212 / Sweden / 02-03), (NCBI-ID 410251); Human respiratory syncytial virus (strain RSP41 / Sweden / 02-03), (NCBI-ID 410234); Human respiratory syncytial virus (strain RSP45 / Sweden / 02-03), (NCBI-ID 410235); Human respiratory syncytial virus (strain RSP56 / Sweden / 02-03), (NCBI-ID 410243); Human respiratory syncytial virus (strain RSP58 / Sweden / 02-03), (NCBI-ID 410236); Human respiratory syncytial virus (strain RSP67 / Sweden / 02-03), (NCBI-ID 410244); Human respiratory syncytial virus (strain RSP94 / Sweden / 02-03) (NCBI-ID 410245); Respiratory syncytial virus isolate RSV Memphis-37, (strain Memphis-37) (NCBI-ID 12814).

[0126] In a preferred embodiment of the invention, at least one antigenic peptide or protein is derived from the respiratory syncytial virus isolate RSV Memphis-37 (strain Memphis-37) (NCBI-ID: 12814). Throughout the present invention, including the information contained in the ST25 sequence listing, the abbreviation "HRSV (Memphis-37)" is used for said particularly preferred RSV virus.

[0127] In a preferred embodiment of the invention, at least one antigenic peptide or protein is derived from Human Respiratory Syncytial Virus A2, Human Respiratory Syncytial Virus (Strain A2) (NCBI-ID: 11259). Throughout the present invention, including the information contained in the ST25 sequence listing, the abbreviation "HRSV(A2)" is used for said particularly preferred RSV virus.

[0128] Those skilled in the art will also understand that the teachings of the present invention can be adapted to use amino acid and nucleic acid sequences from any of the RSV strains provided in List 1 to obtain the RNA constructs, compositions, and vaccines of the present invention.

[0129] In various embodiments, the at least one antigenic peptide or protein can be selected from the fusion protein (F), glycoprotein G, short hydrophobic protein SH, matrix protein M, nucleoprotein N, large polymerase L, M2-1 protein, M2-2 protein, phosphoprotein P, nonstructural protein NS1, or nonstructural protein NS2 of respiratory syncytial virus (RSV), or a fragment, variant, or derivative thereof.

[0130] In a particularly preferred embodiment of the first aspect, the at least one antigenic peptide or protein is derived from the RSV fusion (F) protein. In this regard, the amino acid sequence of the at least one antigenic peptide or protein can be selected from any peptide or protein derived from the RSV fusion protein F, or a fragment, variant, or derivative thereof.

[0131] The RSV F protein is initially expressed (after infection of host cells) as a single polypeptide precursor called the full-length fusion protein F (referred to herein as "F0"). F0 forms a trimer in the endoplasmic reticulum and is processed by cellular / host furin-like proteases at two conserved sites to generate F1, F2, and Pep27 polypeptides. The Pep27 polypeptide is excised and does not form part of the mature F protein. The F2 polypeptide is derived from the N-terminal portion of the F0 precursor and is linked to the F1 polypeptide via two disulfide bonds. The F1 polypeptide is derived from the C-terminal portion of the F0 precursor and anchors the mature F protein to the membrane via a transmembrane domain attached to the cytoplasmic tail. Three F2-F1 heterodimer units ("protomers") assemble to form the mature F protein. Initially, the mature F protein is in a metastable form (referred to herein as the "pre-fusion conformation"). Upon stimulation, it undergoes a large, irreversible structural change (herein referred to as the "post-fusion conformation") that allows the virus to fuse with the target cell membrane.

[0132] Thus, the artificial RNA of the first aspect, preferably suitable for vaccination, encodes at least one antigenic peptide or protein derived from the RSV F protein or a fragment or variant thereof.

[0133] In a preferred embodiment, at least one antigenic peptide or protein derived from the RSV F protein can be derived from any of the amino acid sequences (NCBI protein accession numbers) provided in List 2 below.

[0134] List 2: NCBI protein accession numbers for RSV fusion (F) proteins: Accession numbers: Protein, AJF44801.1, AJF44759.1, AJF44661.1, AJF44602.1, 2207424A, AAB38520.1, AAB38517.1, AAB38519.1, AAB38518.1, AVQ93587.1, AVQ93599.1, AVQ93571.1, AVQ93568.1, AVQ93589.1, AVQ93597.1, AVQ93563.1, AVQ93594.1, AVQ93606.1, AVQ93601.1, AVQ93562.1, AVQ93561.1, AVQ936 07.1, AVQ93588.1, AVQ93575.1, AVQ93468.1, AVQ93467.1, AVQ93590.1, AVQ93552.1, AVQ93556.1, AVQ93471.1, AVQ93458.1, AVQ93494.1, AVQ93470.1 , AVQ93489.1, AVQ93542.1, AVQ93472.1, AVQ93514.1, AVQ93485.1, AVQ93533.1, AVQ93481.1, AVQ93546.1, AVQ93512.1, AVQ93554.1, AVQ93551.1, AVQ 93547.1, AVQ93558.1, AVQ93461.1, AVQ93500.1, AVQ93426.1, AVQ93398.1, AVQ93401.1, AVQ93361.1, AVQ93408.1, AVQ93443.1, AVQ93429.1, AVQ9335 9.1, AVQ93365.1, AVQ93366.1, AVQ93402.1, AVQ93377.1, AVQ93412.1, AVQ93391.1, AVQ93457.1, AVQ93372.1, AVQ93455.1, AVQ93364.1, AVQ93378.1, AVQ93393.1, AVQ93362.1, AVQ93585.1, ART28504.1, AVQ93404.1, AOS49123.1, AOS48496.1, AMT78271.1, AHX57174.1, AHW81390.1, AHV81506.1, AFX6 0128.1, AFX60129.1, AEQ63389.1, ARB66328.1, ANZ80034.1, AMN91253.1, P03420.1, AIO08046.1, NP_056863.1, AFX60234.1, AFX60231.1, AFX60232.1、AFX60222.1、AFX60219.1、AFX60215.1、AFX60214.1、AFX60212.1、AFX60208.1、AFX60202.1、AFX60220.1、AFX60213.1、AFX60190.1、AFX60187.1、AFX60201.1、AFX60173.1、AFX60169.1、AFX60162.1、AFX60156.1、AFX60151.1、AFX60150.1、AFX60148.1、AFX60141.1、AFX60127.1、AFX60137.1、AFX60135.1、AFV46420.1、AFX60200.1、AFV46419.1、AFV46417.1、AFV46413.1、AFV46414.1、AFV46410.1、AFV46403.1、AFV46409.1、AFP99061.1、AFM95400.1、AFV46401.1、AFP99064.1、AFM95376.1、AFX60138.1、AFP99060.1、AFM95365.1、AFM55563.1、AFM55530.1、AFM55442.1、AFM55420.1、AFM55552.1、AFM55365.1、AFP99059.1、AFM95385.1、AFM55354.1、AFM55343.1、AFM55387.1、AFM55299.1、AFM55288.1、AFM55266.1、AFM55277.1、AFM55255.1、AFM55222.1、AFM55211.1、AFI25262.1、AFD34266.1、AFM55332.1、AFD34264.1、AFD34262.1、AFD34265.1、AFD34261.1、AFD34260.1、AFD34259.1、AEQ63641.1、AEQ63487.1、AEQ63520.1、AEQ63378.1、AEQ63367.1、4CCF_A、AEQ63334.1、AEO45949.1、AEO45939.1、AEQ63312.1、AEQ63586.1、AEO45919.1、AEO45909.1、AEO45889.1、AEO45879.1、AEO45869.1、AEO45929.1、AEO45850.1、AEO45859.1、AEQ63444.1、AEO23054.1、AEO23052.1、AEO23051.1、AEC32087.1、ADZ95785.1、AEC32085.1、ADZ95784.1、ADZ95783.1、ADZ95782.1、ADZ95781.1、ADZ95779.1、ADZ95780.1、ADZ95777.1、ADZ95778.1、ADZ95776.1、A DZ95775.1、ACY68435.1、ACO83302.1、ABI35685.1、AFI25251.1、AAX23994.1、AAQ97026.1、AAR14266.1、AAQ97027.1、AAQ97030.1、AAQ97028.1、AAC5 7027.1、AAQ97029.1、AAQ97031.1、AAM68160.1、AAM44851.1、P11209.2、P13843.1、AAO72325.1、AAM68157.1、CAA26143.1、1512372A、AAB86664.1、AA O72324.1、AAB82446.1、AAO72323.1、AAM68154.1、AAA47410.1、P12568.1、ARB07894.1、AGG39517.1、BBC54612.1、BBC54636.1、BBC54627.1、BBC5462 1.1、BBC54570.1、BBC54579.1、BBC54595.1、BBC54555.1、BBC54552.1、BBC54564.1、BBC54581.1、BBC54571.1、BBC54553.1、BBC54565.1、BBC54245.1 、BBC54243.1、BBC54238.1、BBC54242.1、BBC54239.1、BBC54235.1、BBC542 34.1、BBC54236.1、BBC54244.1、BBC54186.1、BBC54178.1、BBC54202.1、BB C54151.1、BBC54142.1、BBC54134.1、BBC54170.1、BBC54210.1、BBC54169.1、BBC54213.1、BBC54203.1、BBC54160.1、BBC54163.1、BBC54215.1、BBC54 156.1、BBC54179.1、BBC54150.1、BBC54207.1、BBC54194.1、BBC54149.1、BBC54138.1、BBC54199.1、BBC54220.1、BBC54181.1、BBC54132.1、BBC54146.1、BBC54122.1、BBC54124.1、BBB35202.1、BBB35201.1、BBB35192.1、BBB35 193.1、BBB35199.1、BBB35184.1、BBB35126.1、BBB35133.1、BBB35130.1、BB B35160.1、BBB35162.1、BBB35181.1、BBB35165.1、BBB35121.1、BBB35138. 1、BBB35176.1、BBB35142.1、BBB35136.1、BBB35153.1、BBB35115.1、BBB351 50.1、BBB35097.1、BBB35109.1、BBB35094.1、BBB35183.1、BBB35104.1、BBB35099.1、BBB35188.1、AKA45871.1、ASV65838.1、AGG39373.1、AII22107.1 、AGG39400.1、AGG39457.1、ARR29240.1、ARR29251.1、ARR29189.1、ARR292 07.1、AUH15164.1、ATV81343.1、AUC68654.1、AUC68577.1、AUC68566.1、AUC 68555.1、AUC68478.1、AUC68445.1、AUC68500.1、AUC68522.1、AUC68291.1 、AUC68149.1、AUC68094.1、AMA67097.1、AMA66866.1、AMA66580.1、AIZ9575 0.1、AIZ95541.1、AIZ95519.1、AHY21419.1、AHY21331.1、AHY21165.1、AHY21143.1、AHY21132.1、AGG39478.1、ATV93506.1、ATV93509.1、AIZ95717.1、 ATV81354.1、ART28317.1、BBA57890.1、BBA57901.1、ASZ70099.1、ART28361.1、AQX36844.1、ASK05520.1、ART28427.1、ART28339.1、ART28297.1、ART2 8328.1, ARQ15966.1, ARN61507.1, ARA15413.1, AGG39394.1, APW78845.1, APW78867.1, APW78900.1, APW78878.1, APW78779.1, APW78702.1, APW78713.1、APW78680.1、APW78658.1、APW78647.1、APW78636.1、AOS48870.1、APW78614.1、AMT78905.1、AMT77402.1、AOZ15479.1、AGN28484.1、AHX57240.1、AHX57031.1、AOS48980.1、AOS48848.1、AOS48815.1、AOS48738.1、AOS49068.1、AOS48727.1、AOS48716.1、AOS48683.1、AOS48551.1、AOS48485.1、AOS48518.1、AOS48441.1、AOS48397.1、AOS48375.1、AOS48353.1、AOS48287.1、AOS48320.1、AOS48265.1、AOS48254.1、AOS48221.1、ANZ79638.1、ALC74025.1、AHV81286.1、AOD40888.1、AOD40516.1、AOD40214.1、AOD40125.1、AOD40104.1、AOD40082.1、AOD39908.1、AJF44826.1、AJF44506.1、AJF44535.1、ANZ80463.1、ANZ80408.1、ANZ80397.1、ANZ80386.1、ANZ80331.1、ANZ80320.1、ANZ80364.1、ANZ80276.1、ANZ80221.1、ANZ80188.1、ANZ80144.1、ANZ80133.1、ANZ80111.1、ANZ80122.1、ANZ80056.1、ANZ80012.1、ANZ80067.1、ANZ79979.1、ANZ79935.1、ANZ79990.1、ANZ79902.1、. ANZ79759.1、ANZ79715.1、ANZ79671.1、AMT79718.1、AMT79586.1、AMT7955 3.1、AMT79542.1、AMT79476.1、AMT79388.1、AMT79201.1、AMT79190.1、AMT 79157.1、AMT79124.1、AMT79091.1、AMT79047.1、AMT79014.1、AMT79003.1、AMT78872.1、AMT78832.1、AMT78689.1、AMT78645.1、AMT78513.1、AMT7848 0.1、AMT78447.1、AMT78392.1、AMT78194.1、AMT78084.1、AMT78051.1、AMT77963.1、AMT77908.1、AMT77776.1、AMT77710.1、AMT77424.1、AMN91385.1、 AMN91264.1、AMN91242.1、AHX57504.1、CUS01881.1、CUS01880.1、CUS0187 7.1、CUS01874.1、CUS01870.1、CUS01875.1、AMA67350.1、AMA67251.1、AMA6 7262.1、AMA67229.1、AMA67196.1、AMA67130.1、AMA67185.1、AMA67163.1、AMA67086.1、AMA67075.1、AMA66998.1、AMA66987.1、AMA66976.1、AMA66965 .1、AMA66921.1、AMA66844.1、AMA66833.1、AMA66624.1、AMA66613.1、AMA6 6591.1、AMA66569.1、AMA66547.1、AMA66503.1、AMA66492.1、AMA66481.1、A MA66448.1、AMA66415.1、AMA66393.1、AMA66360.1、AGG39553.1、AGG39469.1、AJZ70144.1、AJZ70166.1、AJZ70155.1、AJZ70133.1、AJZ70067.1、AJZ70 001.1、AJZ69990.1、AJZ69968.1、AJZ69946.1、AJZ69913.1、AJZ69880.1、AJZ69847.1、AJZ69869.1、AJZ69770.1、AJZ69748.1、AJZ69726.1、AJZ69682.1、AJZ69671.1、AJZ69704.1、AJZ69660.1、AJZ69715.1、AJZ69627.1、AJZ69 616.1、AJZ69638.1、AJO16077.1、AJO16055.1、AKE31881.1、AKE31882.1、AK E31878.1、AJF44835.1、AJF44790.1、AJF44737.1、AJF44716.1、AJF44725.1、AJF44643.1、AJF44628.1、AJF44624.1、AJF44613.1、AJF44566.1、AJF445 55.1、AJF44526.1、AJF44517.1、AKA45882.1、AKA45849.1、AHA83913.1、AGN28715.1、AHA83902.1、AHA83837.1、AHA83826.1、AHA83705.1、AHA83630.1 、AGG39505.1、AIY70242.1、AIY70198.1、AII30203.1、AHY21463.1、AHY213 97.1、AHY21320.1、AHY21298.1、AHY21287.1、AHY21276.1、AHY21254.1、AHY 21199.1、AHY21176.1、AHX57537.1、AHX57471.1、AHX57427.1、AHX57042.1、AIZ95981.1、AIZ95893.1、AIZ95871.1、AIZ95816.1、AIZ95772.1、AIZ9562 9.1、AIZ95673.1、AIZ95596.1、AIZ95585.1、AIZ95552.1、AEQ63553.2、AEQ63542.2、AEQ63575.1、AEQ63531.1、AEQ63498.1、AEQ63422.1、AEQ63411.1、 AHX57570.1、AHX57152.1、AHX57064.1、AHX57009.1、AHX56987.1、AHV8210 0.1、AHV82001.1、AHV81891.1、AHV81880.1、AHV81836.1、AHV81682.1、AHV8 1649.1、AHV81484.1、AHV81462.1、AHV81385.1、AHV81363.1、AHV81330.1、A HV81253.1、AHV81154.1、AHV81122.1、AHV81089.1、AHV81012.1、AHV80957.1、AHV80880.1、AHV80869.1、AHV80836.1、AHV80803.1、AGG39559.1、AGG39562.1、AGG39556.1、AGG39550.1、AGG39547.1、AGG39544.1、AGG39541.1、AG G39529.1、AGG39526.1、AGG39523.1、AGG39514.1、AGG39502.1、AGG39499.1、AGG39496.1、AGG39493.1、AGG39484.1、AGG39490.1、AGG39487.1、AGG394 75.1、AGG39472.1、AGG39466.1、AGG39463.1、AGG39454.1、AGG39442.1、AGG39439.1、AGG39436.1、AGG39415.1、AGG39403.1、AGG39397.1、AGG39391.1 、AGG39379.1、AHL84194.1、AHA84012.1、AHJ60043.1、BAO49770.1、BAO49766.1、BAO49767.1、AHA84034.1、AHA84023.1、AHA83990.1、AHA83957.1、AHA 83924.1, AHA83891.1, AHA83880.1, AHA83782.1, AHA83760.1, AHA83694.1, AGT75357.1, AGN92848.1, AGN28792.1, AGN28781.1, AGN28759.1, AGN2874 8.1、AGN28693.1、AGN28638.1、AGN28627.1、AGN28539.1、AGN28528.1、AGN28462.1、AGN28440.1、AGL96787.1、AGL96786.1、AGL96784.1、AAS93662.1、 AAS93657.1、AAS93656.1、AAS93659.1、AAS93660.1、AAS93663.1、AAS93664.1、AAS93655.1、CUS01869.1、AHG54517.1、ASF87348.1、ASF87341.1、ASF8 7344.1、ASF87351.1、ASF87338.1、ASF87342.1、ASF87352.1、ASF87337.1、ASF87336.1、AEQ98756.1、AEQ98757.1、AEQ98755.1、AEQ98752.1、AEQ98753.1、AEQ98747.1、ASF87325.1、ASF87326.1、5W23_A、5EA3_F、5UDD_A、5EA8_F、AHG54458.1、AEN74947.1、AHG54485.1、AHG54477.1、AHG54451.1、AHG54463.1、AHG54445.1、AEO12131.1、AEN74945.1、AEN74944.1、ASF87335.1、AHG54515.1、AHA61605.1、AHV81660.1、AHG54441.1、or AIY60640.1。

[0135] In the context of the present invention, the terms "RSV F protein," "RSV fusion protein (F)," "RSV F," or "F" can be understood in their broadest sense to refer to F0 (F polypeptide precursor), F1, F2, and Pep27 polypeptides, F2-F1 heterodimers, or mature F protein (including three F2-F1 heterodimers), or any fragments and variants thereof. Thus, the term "peptide or protein derived from the RSV fusion (F) protein" refers to, for example, a peptide, protein, fragment, or variant derived from F0 (F protein polypeptide precursor), F1, F2, and Pep27 polypeptides, F2-F1 heterodimers, or mature F protein. Furthermore, the term "peptide or protein derived from the RSV fusion (F) protein" refers to a peptide, protein, fragment, or variant derived from the "RSV F protein" or "RSV fusion protein (F)" defined above, which can be genetically modified to lack certain protein elements (e.g., cytoplasmic tail, furin cleavage site, Pep27) or to include additional elements (e.g., linker elements, heterologous signal peptides, etc.). For example, the term "peptide or protein derived from the RSV fusion (F) protein" refers to F0, F-del, F0_DSCav1, F_DSCav1_mut1, F_DSCav1_mut2, F_DSCav1_mut3, F_DSCav1_mut4, F_DSCav1_mut5, F_DSCav1_mut6, F_DSCav1_mut7, F_DSCav1_mut8, F_DSCav1_mut0, F-del_DSCav1 ... By this reference is meant peptides, proteins, fragments or variants derived from Cav1_mut1, F-del_DSCav1_mut2, F-del_DSCav1_mut3, F-del_DSCav1_mut4, F-del_DSCav1_mut5, F-del_DSCav1_mut6, F-del_DSCav1_mut7, F-del_DSCav1_mut8, F-del_DSCav1_mut0 (see Table 1 for a description of these constructs).Particularly suitable F protein variants that may be encoded by the RNA of the first aspect are identified below and shown in Table 1.

[0136] When referring to amino acid (aa) residues and their positions in the RSV F protein, any numbering used herein (unless otherwise specified) refers to the position of each aa residue in the corresponding F precursor protein of HRSV(A2) (SEQ ID NO: 68) or the corresponding F precursor protein of HRSV(Memphis-37) (SEQ ID NO: 8937 or 11726), with position "1" corresponding to the first aa residue, i.e., the N-terminal aa residue of the HRSV(A2)F precursor protein or HRSV(Memphis-37)F precursor protein.

[0137] In a preferred embodiment, at least one coding sequence of the RNA of the first aspect encodes at least one antigenic peptide or protein derived from the RSV F protein, wherein the RSV F protein is a full-length F protein (F0) or a C-terminally deleted F protein (F-del), or a fragment or variant thereof.

[0138] In this context, any RSV F full-length protein (precursor protein, referred to as "F0") can be used as a suitable antigen, preferably selected from any of the NCBI protein accession numbers provided in List 2 or from any of SEQ ID NOS: 68, 8279-8967, or 11726. In a preferred embodiment of the present invention, the full-length F protein (F0) (SEQ ID NO: 68) of HRSV (A2) is preferably used (see Table 1). In another preferred embodiment of the present invention, the full-length F protein (F0) (SEQ ID NO: 8937 or 11726) of HRSV (Memphis-37) is preferably used (see Table 1).

[0139] In this regard, any RSV F with a C-terminal deletion (F-del) can be used as a suitable antigen, preferably from any of the NCBI protein accession numbers shown in List 2, or selected from SEQ ID NOs: 483, 8968-9683, or 12095. An example of such a deletion variant is the RSV-Fdel554-574 protein (Oomens et al., 2006. J. Virol. 80(21):10465-77), which removes aa residues 554-574 of the full-length F0 protein. Deleting a significant portion of the cytoplasmic end of F0 (aa 554-574) leads to improved intracellular / cell surface transport in vitro and significantly increases cell surface expression of RSV F. Increased cell surface presentation improves B cell recognition (which is consistent with published data; see WO 2015024668). In a preferred embodiment of the present invention, a C-terminally deleted F protein, referred to herein as "F-del" (SEQ ID NO: 483, 9653, or 12095), is preferably used (see Table 1). In light of the high level of structural conservation of the RSV F protein between different RSV strains (see, e.g., Hause et al., 2017, PLOS ONE 12(6):e0180623), deletion of aa 554-574 is applicable to various RSV F protein sequences of various RSV isolates.

[0140] In a particularly preferred embodiment, the artificial RNA of the first aspect encodes at least one antigenic peptide or protein derived from the RSV F protein, wherein the RSV F protein is designed to stabilize the antigen in a pre-fusion conformation, which is particularly advantageous for an efficient RSV vaccine because several potential epitopes for neutralizing antibodies are accessible in this protein conformation.

[0141] In some embodiments, the RSV F protein comprises one or more amino acid substitutions that stabilize the F protein in a pre-fusion conformation, for example, substitutions that stabilize the membrane distal portion of the F protein (including the N-terminal region of the F1 polypeptide) in the pre-fusion conformation. For example, the amino acid substitutions can introduce non-native disulfide bonds or can be cavity-filling amino acid substitutions.

[0142] Thus, in some embodiments, the preferred RSV F protein comprises S155C and S290C substitutions that form non-native disulfide bonds that stabilize the protein in a pre-fusion conformation (i.e., a conformation that specifically binds to one or more pre-fusion specific antibodies and / or exhibits a preferred antigenic site present in the pre-fusion structure rather than the post-fusion structure of the RSV F protein). In a further embodiment, the recombinant RSV F protein may further comprise an F, L, W, Y, H, or M substitution at position 190, position 207, or positions 190 and 207.

[0143] An example of an RSV F protein designed to stabilize an antigen in a pre-fusion conformation is an RSV F protein containing DSCav1 mutations (S155C, S290C, S190F, and V207L), or a fragment or variant thereof. Such RSV F DSCav1 proteins have been described in the art (WO 2014160463).

[0144] Thus, in a particularly preferred embodiment, the artificial RNA of the first aspect encodes at least one antigenic peptide or protein derived from the RSV F protein, wherein the RSV F protein comprises a DSCav1 mutation (S155C, S290C, S190F, and V207L), or a fragment or variant thereof.

[0145] In the context of the present invention, it should be understood that RSV F can be mutated at positions S155C, S290C, S190F, and V207L ​​to stabilize the protein in the pre-fusion conformation and can be suitably used in the context of the present invention. Thus, any protein selected from any of the NCBI protein accession numbers provided above, or SEQ ID NOs: 68, 8279-8967, 483, 8968-9683, 12095, or 11726, or a fragment or variant thereof, can be selected by those skilled in the art to introduce amino acid changes due to S155C, S290C, S190F, and V207L ​​to generate various RSV F DSCav1 proteins.

[0146] In a preferred embodiment, the RSV F full-length protein (precursor protein, "F0") of HRSV (A2) (SEQ ID NO: 68) is used and amino acid changes S155C, S290C, S190F, and V207L ​​are introduced to yield the amino acid sequence of SEQ ID NO: 898. This RSV F protein is referred to herein as "FO_DSCav1" throughout the present invention (see Table 1 (Preferred RSV F Protein Antigen Design)).

[0147] In another preferred embodiment, the RSV F_del protein (deleted at the cytoplasmic tail (aa 554-574)) of HRSV (A2) (SEQ ID NO: 483) is used and the S155C, S290C, S190F, and V207L ​​amino acid changes are introduced to produce the amino acid sequence of SEQ ID NO: 1267. This RSV F protein is referred to herein as "F-del_DSCav1" throughout the present invention (see Table 1 (Preferred RSV F Protein Antigen Design)).

[0148] In a preferred embodiment, the RSV F full-length protein (precursor protein, "F0") (SEQ ID NO: 8937 or 11726) of HRSV (Memphis-37) is used to introduce amino acid changes S155C, S290C, S190F, and V207L, resulting in the amino acid sequence of SEQ ID NO: 16464. This RSV F protein is referred to herein as "FO_DSCav1" throughout the present invention (see Table 1 (Preferred RSV F Protein Antigen Design)).

[0149] In another preferred embodiment, the RSV F_del protein (cytoplasmic tail (aa 554-574) deleted) (SEQ ID NO: 9653 or 12095) of HRSV (Memphis-37) is used to introduce amino acid changes S155C, S290C, S190F, and V207L, resulting in the amino acid sequence of SEQ ID NO: 12833. This RSV F protein is referred to herein as "F-del_DSCav1" throughout the present invention (see Table 1 (Preferred RSV F Protein Antigen Design)).

[0150] In a preferred embodiment, the at least one antigenic peptide or protein can be an engineered protein containing two subunits of mature F, F1 and F2, as a single polypeptide chain, where F2 and F1 are preferably connected via a linker (GS). Examples of engineered F2-linker-F1 fusion proteins (e.g., F(1-103)-GS-F(145-574) or F(1-103)-GS-F(145-553)) have been described in the art (Joyce, M. Gordon et al., "Iterative structure-based improvement of a fusion-glycoprotein vaccine against RSV," Nature structural & molecular biology 23.9 (2016): 811; WO 2017172890). The F2-linker-F1 RSV F protein does not contain aa 104 to 144 (including the furin cleavage site and Pep27) and contains a linker element (e.g., a GS linker) between the F2 polypeptide and the F1 polypeptide. The F2-linker-F1 protein can exhibit excellent properties in terms of stability and / or antigenicity.

[0151] Thus, in a preferred embodiment, the RSV F protein comprises two subunits, F2 and F1, fused to a single polypeptide chain, wherein F2 and F1 are connected via a linker element, preferably a GS linker as defined herein, to preferably generate a stable F2-linker-F1 protein.

[0152] Preferably, the F2-linker-F1 fusion protein, e.g., F(1-103)-GS-F(145-574) or F(1-103)-GS-F(145-553), further comprises a DScav1 mutation as outlined above (referred to herein as "mut0"; e.g., SEQ ID NOs: 3850, 4219 or 13940, 14309).

[0153] In a particularly preferred embodiment, the RSV F protein is selected from the group consisting of (S46G, A149C, S215P, Y458C, K465Q), (S46G, E92D, A149C, S215P, Y458C, K465Q), (S46G, N67I, E92D, A149C, S215P, Y458C, K465Q), (A149C, Y458C), (N183GC, N428C), (Q98C, Q361C, S46G, E The amino acid sequence of the present invention may further comprise at least one additional mutation selected from (Q98C, Q361C, L95M, I221M, R429K, K465Q), (Q98C, Q361C, L95M, I221M, R429K), or (N183GC, N428C, S46G, N67I, E92D, S215P, K465Q), or a fragment or variant thereof.

[0154] In a particularly preferred embodiment, the F2-linker-F1 protein (F(1-103)-GS-F(145-574) or F(1-103)-GS-F(145-553)) preferably contains, in addition to the DSCav1 mutation, S46G, A149C, S215P, Y458C, K465Q (referred to herein as "mut1"; e.g., SEQ ID NOs: 1636, 2005, or 14678, 15047); or S46G, E92D, A149C, S215P, Y458C , K465Q (referred to herein as "mut2"; e.g., SEQ ID NOs: 2374, 2743, or 15416, 15785); or S46G, N67I, E92D, A149C, S215P, Y458C, K465Q (referred to herein as "mut3"; e.g., SEQ ID NOs: 3112, 3481, or 13202, 13571), or any fragment or variant thereof (see Table 1 (Preferred RSV F protein antigen design)).

[0155] In another embodiment, the F2-linker-F1 protein (F(1-103)-GS-F(145-574) or F(1-103)-GS-F(145-553)) preferably contains, in addition to the DSCav1 mutation, A149C, Y458C (referred to herein as "mut4"; e.g., SEQ ID NOs: 4588, 4957, or 16154, 16523); or N183GC, N428C (referred to herein as "mut5"; e.g., SEQ ID NOs: 5326, 5695, or 16892, 17261); or Q98C, Q361C, S46G, E92D, L95M, S215P, I217P, I22 or Q98C, Q361C, L95M, I221M, R429K (referred to herein as "mut7"; e.g., SEQ ID NOs: 6802, 7171 or 18368, 18737); or N183GC, N428C, S46G, N67I, E92D, S215P, K465Q (referred to herein as "mut8"; e.g., SEQ ID NOs: 7540, 7909 or 19106, 19475), or a fragment or variant thereof.

[0156] In the context of the present invention, any RSV F can be adapted so that the two subunits F1 and F2 are contained in a single polypeptide chain (F2 and F1 can be linked via a linker, e.g., a (GS) linker for increasing protein stability, as described for the mutations "mut1", "mut2", and "mut3"). For example, by deleting aa 104-144 of the F0 polypeptide chain (described above), introducing a linker element between F2 and F1, as described above, and introducing a suitable amino acid substitution as described above. Thus, any protein selected from the NCBI protein accession numbers provided above (see List 2), or SEQ ID NOs: 68, 8279-8967, 483, 8968-9683, 11726, 12095, or a fragment or variant thereof, can be adapted by one of skill in the art to generate an F2-linker-F1 fusion protein as outlined herein, including (S46G, A149C, S215P, Y458C, K465Q), (S46G, E92D, A149C, S215P, Y458C, K465Q), (S46G, E92D, A149C, S215P, Y458C, K465Q), Further adaptations can be achieved by introducing aa substitutions and / or DSCav1 mutations: (S46G, N67I, E92D, A149C, S215P, Y458C, K465Q), (A149C, Y458C), (N183GC, N428C), (Q98C, Q361C, S46G, E92D, L95M, S215P, I217P, I221M, R429K, K465Q), (Q98C, Q361C, L95M, I221M, R429K), or (N183GC, N428C, S46G, N67I, E92D, S215P, K465Q). Furthermore, apart from the use of the GS linker outlined above, those skilled in the art will appreciate that various known linker elements can be selected from to arrive at similar, similarly suitable RSV F protein variants (e.g., SEQ ID NOS: 117-162 of Domestic Publication No. 2017 / 172890, or fragments or variants thereof, or SEQ ID NOS: 1509-1565 of Domestic Publication No. 2017 / 081082, or fragments or variants thereof).

[0157] In a preferred embodiment, the F2-linker-F1 protein (F(1-103)-GS-F(145-574) or F(1-103)-GS-F(145-553)) comprises the DSCav-1 mutations outlined above (referred to herein as F_DSCav1_mut0 or F-del_DSCav1_mut0). In a preferred embodiment, the F2-linker-F1 protein (F(1-103)-GS-F(145-574) or F(1-103)-GS-F(145-553)) further comprises the DSCav-1 mutations outlined above and the amino acid substitution mut1 as defined above (referred to herein as F_DSCav1_mut1 or F-del_DSCav1_mut1). In a preferred embodiment, the F2-linker-F1 protein (F(1-103)-GS-F(145-574) or F(1-103)-GS-F(145-553) GS-F(145-574) or F(1-103)-GS-F(145-553) further comprises the DSCav-1 mutations outlined above and the amino acid substitution mut2 as defined above (referred to herein as F_DSCav1_mut2 or F-del_DSCav1_mut2). In a preferred embodiment, the F2-linker-F1 protein (F(1-103)-GS-F(145-574) or F(1-103)-GS-F(145-553) comprises the DSCav-1 mutations outlined above and the amino acid substitution mut2 as defined above. In a preferred embodiment, the F2-linker-F1 protein (F(1-103)-GS-F(145-574) or F(1-103)-GS-F(145-553)) further comprises the DSCav-1 mutations outlined above and the amino acid substitution mut4 as defined above (referred to herein as F_DSCav1_mut3 ​​or F-del_DSCav1_mut3). In a preferred embodiment, the F2-linker-F1 protein (F(1-103)-GS-F(145-574) or F(1-103)-GS-F(145-553) further comprises the DSCav-1 mutations outlined above and the amino acid substitution mut5 as defined above (herein referred to as F_DSCav1_mut5 or F-del_DSCav1_mut5).In a preferred embodiment, the F2-linker-F1 protein (F(1-103)-GS-F(145-574) or F(1-103)-GS-F(145-553) comprises the DSCav-1 mutations outlined above and further comprises the amino acid substitution mut6 as defined above (referred to herein as F_DSCav1_mut6 or F-del_DSCav1_mut6). In a preferred embodiment, the F2-linker-F1 protein (F(1-103)-GS-F(145-574) or F(1-103)-GS-F(145-553) comprises the DSCav-1 mutations outlined above and further comprises the amino acid substitution mut6 as defined above (referred to herein as F_DSCav1_mut6 or F-del_DSCav1_mut6). In a preferred embodiment, the F2-linker-F1 protein (F(1-103)-GS-F(145-574) or F(1-103)-GS-F(145-553)) further comprises the DSCav-1 mutations outlined above and the amino acid substitution mut7 as defined above (referred to herein as F_DSCav1_mut7 or F-del_DSCav1_mut7). In a preferred embodiment, the F2-linker-F1 protein (F(1-103)-GS-F(145-574) or F(1-103)-GS-F(145-553)) further comprises the DSCav-1 mutations outlined above and the amino acid substitution mut8 as defined above (referred to herein as F_DSCav1_mut8 or F-del_DSCav1_mut8).

[0158] A detailed description of particularly preferred RSV F proteins is set forth in Table 1.

[0159] In Table 1, all references to amino acid (aa) residues and their positions in the RSV F protein refer to the position of each aa residue in the corresponding F0 precursor protein of HRSV (A2) (SEQ ID NO: 68) or HRSV (Memphis-37) (SEQ ID NO: 8937 or 11726). Additionally, the preferred RSV in Table 1 Abbreviations for F protein antigen designs are used throughout the description of the present invention (e.g., "F0", "F-del", "F0_DSCav1", "F-del_DSCav1", "F_DSCav1_mut1", "F-del_DSCav1_mut1", "F_DSCav1_mut2", "F-del_DSCav1_mut2", "F_DSCav1_mut3", "F-del_DSCav1_mut3", "F_DSCav1_mut4", "F-del_DSCav1_mut4", "F_DSCav1_mut5", "F-del_DSCav1_mut5", "F_DSCav1_mut6", "F-del_DSCav1_mut6", "F_DSCav1_mut7", "F-del_DSCav1_mut7", "F_DSCav1_mut8", "F-del_DSCav1_mut8", "F_DSCav1_mut0", "F-del_DSCav1_mut0"). Column A of Table 1 provides the protein sequence numbers of each RSV F protein antigen design derived from HRSV (A2); Column B of Table 1 provides the protein sequence numbers of each RSV F protein antigen design derived from HRSV (Memphis-37). Note that the description of the present invention is based on the ST25 sequence listing of the present application. <223> Explicitly include the information provided under the identifier.

[0160] [Table 1-1] [Table 1-2]

[0161] In a particularly preferred embodiment, the artificial RNA according to the first aspect encodes at least one antigenic peptide or protein derived from the RSV fusion (F) protein, The F protein is selected from F0, F-del, F0_DSCav1, F_DSCav1_mut0, F_DSCav1_mut1, F_DSCav1_mut2, F_DSCav1_mut3, F_DSCav1_mut4, F_DSCav1_mut5, F_DSCav1_mut6, F_DSCav1_mut7, F_DSCav1_mut8, F-del_DSCav1, F-del_DSCav1_mut0, F-del_DSCav1_mut1, F-del_DSCav1_mut2, F-del_DSCav1_mut3, F-del_DSCav1_mut4, F-del_DSCav1_mut5, F-del_DSCav1_mut6, F-del_DSCav1_mut7, F-del_DSCav1_mut8 (see, e.g., Table 1), or a fragment or variant thereof.

[0162] Particularly preferred and advantageous in the present invention are RSV F proteins selected from F-del_DSCav1, F-del_DSCav1_mut0, F-del_DSCav1_mut1, F-del_DSCav1_mut2, F-del_DSCav1_mut3, F-del_DSCav1_mut4, F-del_DSCav1_mut5, F-del_DSCav1_mut6, F-del_DSCav1_mut7, F-del_DSCav1_mut8 (see, for example, Table 1), or fragments or variants thereof.

[0163] In a preferred embodiment, the artificial RNA of the first aspect is selected from the group consisting of SEQ ID NOs: 68, 483, 898, 1267, 1636, 2005, 2374, 2743, 3112, 3481, 3850, 4219, 4588, 4957, 5326, 5695, 6064, 6433, 6802, 7171, 7540, 7909, 11726, 12095, 12464, 12833, 13940, 14309, 14678, 15047, 15416, 15785, 13202, 13571, 16154, 16523, 16892, 17261, 1763 0, 17999, 18368, 18737, 19106, 19475 (see, e.g., Table 1), or 8279-9683 (SEQ ID NO: 1), or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any of these sequences, or a fragment or variant of any of these sequences. Further information regarding each of these preferred amino acid sequences encoding RSV proteins can be found in the Sequence Listing, particularly those identified by the identifiers 0, 17999, 18368, 18737, 19106, 19475 (see, e.g., Table 1), or 8279-9683 (SEQ ID NO: 1), or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical thereto. <223> This can also be obtained from the details given below.

[0164] In a preferred embodiment, the artificial RNA of the first aspect is selected from the group consisting of SEQ ID NOs: 68, 483, 898, 1267, 1636, 2005, 2374, 2743, 3112, 3481, 3850, 4219, 4588, 4957, 5326, 5695, 6064, 6433, 6802, 7171, 7540, 7909, 11726, 12095, 12464, 12833, 13940, 14309, 14678, 15047, 15416, 15785, 13202, 13571, 16154, 16523, 16892, 1726 1, 17630, 17999, 18368, 18737, 19106, 19475 (see, e.g., Table 1), or at least one amino acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any of these sequences, or a fragment or variant of any of these sequences. Further information regarding each of these preferred amino acid sequences encoding RSV proteins can be found in the Sequence Listing, particularly those identified by the identifiers 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84 <223> This can also be obtained from the details given below.

[0165] In a particularly preferred embodiment, the artificial RNA of the first aspect comprises at least one coding sequence encoding at least one antigenic peptide or protein comprising or consisting of at least one amino acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any of SEQ ID NOs: 1267, 2005, 2743, 3481, 4219, 4957, 5695, 6433, 7171, 7909, 12833, 14309, 15047, 15785, 13571, 16523, 17261, 17999, 18737, 19475 (see, e.g., Table 1), or a fragment or variant of any of these sequences. Further information regarding each of these preferred amino acid sequences encoding RSV proteins can be found in the Sequence Listing, particularly the identifiers AA, ... <223> This can also be obtained from the details given below.

[0166] In other embodiments, the artificial RNA of the first aspect comprises at least one coding sequence encoding at least one antigenic peptide or protein comprising or consisting of at least one amino acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any of SEQ ID NOs: 8279 to 9683, or a fragment or variant of any of these sequences. Further information regarding each of these preferred amino acid sequences encoding RSV proteins can be found in the Sequence Listing, in particular, by reference to the identifiers 8279-9683, as described below. <223> This can also be obtained from the details given below.

[0167] In another embodiment, the artificial RNA defined herein comprises at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV, comprising or consisting of at least one amino acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any of SEQ ID NOs: 1-1428 of WO 2014 / 160463, or a fragment or variant of any of these sequences. In this context, SEQ ID NOs: 1-1428 of WO 2014 / 160463 and the related disclosures thereof are incorporated herein by reference.

[0168] In other embodiments, the artificial RNA defined herein comprises at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV, comprising or consisting of at least one amino acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any of SEQ ID NOS: 1-11 of WO 2015 / 024668, or a fragment or variant of any of these sequences. In this context, SEQ ID NOS: 1-11 of WO 2015 / 024668 and the related disclosures thereof are incorporated herein by reference.

[0169] In other embodiments, the artificial RNA defined herein comprises at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV that is identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any of SEQ ID NOs: 3, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 243, or 245 of WO 2017 / 070622, or a fragment or variant of any of these sequences. In this context, SEQ ID NOs: 3, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 243, or 245 of WO 2017 / 070622 and the disclosures related thereto are incorporated herein by reference.

[0170] In other embodiments, the artificial RNA defined herein comprises at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV that comprises or consists of at least one amino acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any of SEQ ID NOS: 1-65, 81-95, 110-116 of WO 2017 / 172890, or a fragment or variant of any of these sequences. In this context, SEQ ID NOS: 1-65, 81-95, 110-116 of WO 2017 / 172890 and the related disclosures thereof are incorporated herein by reference.

[0171] According to another preferred embodiment, the artificial RNA of the invention encodes at least one antigenic peptide or protein as defined above and may additionally encode at least one further heterologous peptide or protein element.

[0172] Preferably, at least one additional peptide or protein element can facilitate secretion of the encoded antigenic peptide or protein of the invention (e.g., via a secretory signal sequence), facilitate binding of the encoded antigenic peptide or protein of the invention to the plasma membrane (e.g., via a transmembrane element), facilitate formation of an antigen complex (e.g., via a multimerization domain), or facilitate formation of a virus-like particle (VLP-forming sequence). Furthermore, the artificial nucleic acid sequence of the invention can further encode a peptide linker element, a self-cleaving peptide, an immunological adjuvant sequence, or a dendritic cell targeting sequence. Suitable multimerization domains can be selected from the list of amino acid sequences according to SEQ ID NOs: 1116 to 1167 of WO 2017 / 081082, or fragments or variants of these sequences. Trimerization and tetramerization elements can be selected, for example, from engineered leucine zippers (engineered α-helical coiled-coil peptides that adopt a parallel trimer state), fibritin foldon domains derived from enterobacterial phages T4, GCN4pII, GCN4-pLI, and p53. In this context, fibritin foldon domains from enterobacterial phages T4, GCN4pII, GCN4-pLI, and p53 are preferred. Suitable transmembrane elements can be selected from the list of amino acid sequences according to SEQ ID NOs: 1228 to 1343 of WO 2017 / 081082, or fragments or variants of these sequences. Suitable VLP-forming sequences can be selected from the list of amino acid sequences according to SEQ ID NOs: 1168 to 1227 of WO 2017 / 081082, or fragments or variants of these sequences. Suitable peptide linkers can be selected from the list of amino acid sequences according to SEQ ID NOs: 1509 to 1565 of WO 2017 / 081082, or fragments or variants of these sequences. Suitable self-cleaving peptides can be selected from the list of amino acid sequences according to SEQ ID NOs: 1434 to 1508 of WO 2017 / 081082, or fragments or variants of these sequences.Suitable immunological adjuvant sequences can be selected from the list of amino acid sequences set forth in SEQ ID NOS: 1360-1421 of WO 2017 / 081082, or fragments or variants of these sequences. Suitable dendritic cell (DC) targeting sequences can be selected from the list of amino acid sequences set forth in SEQ ID NOS: 1344-1359 of WO 2017 / 081082, or fragments or variants of these sequences. Suitable secretory signal peptides can be selected from the list of amino acid sequences set forth in SEQ ID NOS: 1-1115 and 1728 of WO 2017 / 081082, or fragments or variants of these sequences. At the nucleic acid level, any nucleic acid sequence (e.g., RNA sequence) encoding such an amino acid sequence can be selected. In this context, the disclosure of WO 2017 / 081082 is incorporated herein by reference. A heterologous secretory signal sequence can increase secretion of the encoded antigenic peptide or protein.

[0173] According to embodiments, the secretory signal sequence comprises an amino acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any of SEQ ID NOs: 21329-21362, or a fragment or variant of these sequences. Further information regarding each of these preferred amino acid sequences encoding secretory signal sequences can be found in the Sequence Listing, in particular the identifiers <223> This can also be obtained from the details given below.

[0174] According to a preferred embodiment, the artificial nucleic acid, particularly the artificial RNA, comprises at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV, preferably derived from the RSV F protein, as defined herein, or fragments and variants thereof. In this context, any coding sequence encoding at least one antigenic peptide or protein derived from RSV, preferably derived from the RSV F protein, or fragments and variants thereof, can be understood as a suitable coding sequence and can therefore be included in the artificial RNA of the first aspect.

[0175] In a preferred embodiment, the artificial RNA of the first aspect comprises at least one coding sequence encoding at least one antigenic peptide or protein derived from the RSV F protein defined herein, preferably SEQ ID NOs: 68, 483, 898, 1267, 1636, 2005, 2374, 2743, 3112, 3481, 3850, 4219, 4588, 4957, 5326, 5695, 6064, 6433, 6802, 7171, 7540, 7909, 11726, 120 95, 12464, 12833, 13940, 14309, 14678, 15047, 15416, 15785, 13202, 13571, 16154, 16523, 16892, 17261, 17630, 17999, 18368, 18737, 19106, 19475, or a variant thereof.At the nucleic acid level, any nucleic acid sequence, in particular SEQ ID NOs: 68, 483, 898, 1267, 1636, 2005, 2374, 2743, 3112, 3481, 3850, 4219, 4588, 4957, 5326, 5695, 6064, 6433, 6802, 7171, 7540, 7909, 11726, 12095, 12464, 12833, 13940, 14309, 14678, 15047, 15416, 15785, 13940, 14309, 14678, 15047, 15416, 15785, 13940, 14509, 14678, 15509, 15609, 15709, 15809, 15909, 16009, 16109, 16209, 16309, 16409, 16509, 16609, 16709, 16809, 16909, 16910, 16920, 16920, 16930, 16940, 1695 ... 202, 13571, 16154, 16523, 16892, 17261, 17630, 17999, 18368, 18737, 19106, 19475, or any RNA sequence encoding a fragment or variant thereof, or SEQ ID NOs: 68, 483, 898, 1267, 1636, 2005, 2374, 2743, 3112, 3481, 3850, 4219, 4588, 4957, 5326 , 5695, 6064, 6433, 6802, 7171, 7540, 7909, 11726, 12095, 12464, 12833, 13940, 14309, 14678, 15047, 15416, 15785, 13202, 13571, 16154, 16523, 16892, 17261, 17630, 17999, 18368, 18737, 19106, 19475, and 8 It should be understood that any nucleic acid sequence (e.g., DNA sequence, RNA sequence) that encodes an amino acid sequence that is 6%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical, or a fragment or variant thereof, can be selected and therefore can be understood as a suitable coding sequence and therefore can be encompassed by the artificial RNA of the first aspect of the present invention.

[0176] In other embodiments, the artificial RNA of the first aspect can comprise or consist of at least one coding sequence encoding any of SEQ ID NOs: 8279 to 9683 or a fragment of a variant thereof. At the nucleic acid level, it should be understood that any nucleic acid sequence, in particular any RNA sequence encoding an amino acid sequence identical to SEQ ID NOs: 8279 to 9683, or a fragment or variant thereof, or any nucleic acid sequence (e.g., DNA sequence, RNA sequence) encoding an amino acid sequence at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any of SEQ ID NOs: 8279 to 9683, or a fragment or variant thereof, can be selected and can therefore be understood as a suitable coding sequence and can therefore be encompassed by the artificial RNA of the first aspect of the present invention.

[0177] In another embodiment, the artificial RNA of the first aspect may comprise or consist of at least one coding sequence encoding any of SEQ ID NOs: 1 to 1428 of WO 2014 / 160463 or a fragment of a variant thereof. At the nucleic acid level, it should be understood that any nucleic acid sequence, in particular any RNA sequence that encodes an amino acid sequence identical to SEQ ID NOs: 1 to 1428 of WO 2014 / 160463, or a fragment or variant thereof, or any nucleic acid sequence (e.g., DNA sequence, RNA sequence) that encodes an amino acid sequence that is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any of SEQ ID NOs: 1 to 1428 of WO 2014 / 160463, or a fragment or variant thereof, can be selected and can therefore be understood as a suitable coding sequence and can therefore be encompassed by the artificial RNA of the first aspect of the present invention.

[0178] In other embodiments, the artificial RNA of the first aspect can comprise or consist of at least one coding sequence encoding any of SEQ ID NOS: 1-11 of WO 2015 / 024668 or a fragment of a variant thereof. At the nucleic acid level, it should be understood that any nucleic acid sequence, in particular any RNA sequence encoding an amino acid sequence identical to SEQ ID NOS: 1-11 of WO 2015 / 024668, or a fragment or variant thereof, or any nucleic acid sequence (e.g., DNA sequence, RNA sequence) encoding an amino acid sequence at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any of SEQ ID NOS: 1-11 of WO 2015 / 024668, or a fragment or variant thereof, can be selected and can therefore be understood as a suitable coding sequence and can therefore be encompassed by the artificial RNA of the first aspect of the present invention.

[0179] In other embodiments, the artificial RNA of the first aspect can comprise or consist of at least one coding sequence encoding any of SEQ ID NOs: 3, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 243, or 245 of WO 2017 / 070622, or a fragment of a variant thereof. At the nucleic acid level, any nucleic acid sequence, in particular any RNA sequence that encodes an amino acid sequence that is identical to SEQ ID NO: 3, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 243 or 245 of WO 2017 / 070622, or a fragment or variant thereof, or any of SEQ ID NO: 3, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 243 or 245 of WO 2017 / 070622. It should be understood that any nucleic acid sequence (e.g., DNA sequence, RNA sequence) that encodes an amino acid sequence that is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical, or a fragment or variant thereof, can be selected and therefore can be understood as a suitable coding sequence and therefore can be encompassed by the artificial RNA of the first aspect of the present invention.

[0180] In another embodiment, the artificial RNA of the first aspect may comprise or consist of at least one coding sequence encoding any of SEQ ID NOs: 1 to 65, 81 to 95, and 110 to 116 of WO 2017 / 172890, or a fragment of a variant thereof. At the nucleic acid level, it should be understood that any nucleic acid sequence can be selected, in particular any RNA sequence encoding an amino acid sequence identical to SEQ ID NOS: 1-65, 81-95, 110-116 of WO 2017 / 172890, or a fragment or variant thereof, or any nucleic acid sequence (e.g., DNA sequence, RNA sequence) encoding an amino acid sequence that is at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any of SEQ ID NOS: 1-65, 81-95, 110-116 of WO 2017 / 172890, or a fragment or variant thereof, and can therefore be understood as a suitable coding sequence, and therefore can be encompassed by the artificial RNA of the first aspect of the present invention.

[0181] Suitably, in a particularly preferred embodiment, the artificial RNA of the first aspect comprises a coding sequence located between the 5'-UTR and the 3'-UTR, preferably downstream of the 5'-UTR and upstream of the 3'-UTR.

[0182] In a preferred embodiment, the artificial RNA of the first aspect is selected from the group consisting of SEQ ID NOs: 69 to 482, 484 to 897, 899 to 1266, 1268 to 1635, 1637 to 2004, 2006 to 2373, 2375 to 2742, 2744 to 3111, 3113 to 3480, 3482 to 3849, 3851 to 4218, 4220 to 4587, 4589 to 4956, 4958 to 5325, 5327 ~5694, 5696~6063, 6065~6432, 6434~6801, 6803~7170, 7172~7539, 7541~7908, 7910~8277, 8278, 11727~12094, 12096~12463, 12465~12832, 12834~13201, 13941~14308, 14310~14677, 14679~15046, 1504 8~15415, 15417~15784, 15786~16153, 13203~13570, 13572~13939, 16155~16522, 16524~16891, 16893~17260, 17262~17629, 17631~17998, 18000~17998, 18369~18736, 18738~19105, 19107~19474, 19476 9, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any of the nucleic acid sequences listed in Tables 1-6, 19843, 21363, 21706, 21843, 21844, 21845, 21846, 21847, 21848, 21849, 21850, 21851, 21852, 21853, 21854, 21855, 21856, 21857, 21858, 21859, 21860, 21861, 21862, 21863, 21864, 21865, 21866, 21867, 21868, 21869, 21870, 21871, 21872, 21873, 21874, 21875, 21876, 21877, 21878, 21879 ...4, 21875, 21876, 21877, 21878, 21879, 21871, 21871, 21872, 21873, 21874, 21875, 21876, 21877 <223> This can also be obtained from the details given below.

[0183] In a particularly preferred embodiment, the artificial RNA of the first aspect is selected from the group consisting of SEQ ID NOs: 69 to 77, 484 to 492, 899 to 906, 1268 to 1275, 1637 to 1644, 2006 to 2013, 2375 to 2382, 2744 to 2751, 3113 to 3120, 3482 to 3489, 3851 to 3858, 4220 to 4227, 4589 to 4596, 4958 to 4965, 5327 to 5328, 5429 to 5430, 5520 to 5531, 5521 to 5522, 5523 to 5524, 5525 to 5532, 5526 to 5533, 5527 to 5534, 5528 to 5535, 5529 to 5536, 5537 to 5538, 5549 to 5551, 5549 to 5552, 5553 to 5554, 5555 to 5556, 5557 to 5558, 5559 to 5560, 5561 to 5562, 5563 to 5564, 5570 to 5572, 5573 to 5576, 5580 to 5582, 5581 to 5583, 5582 to 5584, 5585 to 5585, 5596 to 5598, 5601 to 5602, 334, 5696~5703, 6065~6072, 6434~6441, 6803~6810, 7172~7179, 7541~7548, 7910~7917, 21363~21384, 11727~11734, 12096~12103, 12465~12472, 12834~12841, 13941~13948, 14310~14317, 14679~14686, 1 5048~15055, 15417~15424, 15786~15793, 13203~13210, 13572~13579, 16155~16162, 16524~16531, 16893~16900, 17262~17269, 17631~17638, 18000~18007, 18369~18376, 18738~18745, 19107~19114, 1947 6-19483, 21389-21410, or a fragment, or fragment or variant of any of these sequences (see also Tables 3 and 4). Further information regarding each of these preferred amino acid sequences can be found in the Sequence Listing, in particular the identifiers <223> This can also be obtained from the details given below.

[0184] In other embodiments, the artificial RNA of the first aspect comprises a coding sequence comprising at least one nucleic acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NOs: 383 to 388 of WO 2014 / 160463, or a fragment or variant of any of these sequences.

[0185] In other embodiments, the artificial RNA of the first aspect comprises a coding sequence comprising at least one nucleic acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NOs: 12 to 22 of WO 2015 / 024668, or a fragment or variant of any of these sequences.

[0186] In other embodiments, the artificial RNA of the first aspect comprises a coding sequence comprising at least one nucleic acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NOs: 1, 2, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 242, 244, 246, 257, 258-280 of WO 2017 / 070622, or a fragment or variant of any of these sequences.

[0187] In other embodiments, the artificial RNA of the first aspect comprises a coding sequence comprising at least one nucleic acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NOs: 96-99 of WO 2017 / 172890, or a fragment or variant of any of these sequences.

[0188] According to a preferred embodiment, the artificial RNA is a modified and / or stabilized artificial RNA.

[0189] Thus, according to a preferred embodiment, the artificial RNA of the present invention can be provided as a "stabilized artificial RNA," i.e., an RNA that exhibits improved resistance to degradation in vivo, and / or an artificial RNA that exhibits improved stability in vivo, and / or an artificial RNA that exhibits improved translatability in vivo. Specific preferred modifications in this context that are suitable for "stabilizing" an artificial RNA are described below.

[0190] Such stabilization can be achieved by providing "dried RNA" and / or "purified RNA" as specified herein. Alternatively or additionally, such stabilization can be achieved, for example, by a modified phosphate backbone of the artificial RNA of the present invention. Backbone modifications relevant to the present invention are modifications in which the phosphate of the backbone of nucleotides contained in RNA is chemically modified. Nucleotides that can be preferably used in this context include, for example, phosphorothioate-modified phosphate backbones (preferably, at least one phosphate oxygen contained in the phosphate backbone is replaced with a sulfur atom). Stabilized RNA can also include, for example, nonionic phosphate analogs, such as alkyl and aryl phosphonates in which the charged phosphonate oxygen is replaced with an alkyl or aryl group, or alkyl phosphotriesters and phosphodiesters in which the charged oxygen residue exists in an alkylated form. Such backbone modifications typically include, but are not limited to, modifications from the group consisting of methyl phosphonate, phosphoramidate, and phosphorothioate (e.g., cytidine-5'-O-(1-thiophosphate)).

[0191] Below, we describe suitable modifications that can "stabilize" the artificial RNA of the present invention.

[0192] According to an embodiment, the artificial RNA of the present invention is a modified artificial RNA, and the modification relates to a chemical modification including a backbone modification and a sugar modification or a base modification.

[0193] In this context, the modified artificial RNA defined herein may contain nucleotide analogs / modifications, such as backbone modifications, sugar modifications, or base modifications. A backbone modification relevant to the present invention is a modification in which the phosphate of the backbone of a nucleotide contained in a nucleic acid, e.g., an artificial RNA, is chemically modified. A sugar modification relevant to the present invention is a chemical modification of the sugar of a nucleotide of an RNA nucleotide defined herein. Furthermore, a base modification relevant to the present invention is a chemical modification of the base moiety of a nucleotide of an RNA nucleotide. In this context, the nucleotide analog or modification is preferably selected from nucleotide analogs applicable to transcription and / or translation.

[0194] In a particularly preferred embodiment of the present invention, nucleotide analogues / mutations that can be introduced into modified nucleic acids or in particular modified RNAs as described herein are 2-amino-6-chloropurine riboside-5'-triphosphate, 2-aminopurine-riboside-5'-triphosphate; 2-aminoadenosine-5'-triphosphate, 2'-amino-2'-deoxycytidine-triphosphate, 2-thiocytidine-5'-triphosphate, 2-thiouridine-5'-triphosphate, 2'-fluorothymidine-5'-triphosphate, 2'-O- Methyl-inosine-5'-triphosphate, 4-thiouridine-5'-triphosphate, 5-aminoallylcytidine-5'-triphosphate, 5-aminoallyluridine-5'-triphosphate, 5-bromocytidine-5'-triphosphate, 5-bromouridine-5'-triphosphate, 5-bromo-2'-deoxycytidine-5'-triphosphate, 5-bromo-2'-deoxyuridine-5'-triphosphate, 5-iodocytidine-5'-triphosphate, 5-iodo-2'-deoxycytidine-5'-triphosphate ate, 5-iodouridine-5'-triphosphate, 5-iodo-2'-deoxyuridine-5'-triphosphate, 5-methylcytidine-5'-triphosphate, 5-methyluridine-5'-triphosphate, 5-propynyl-2'-deoxycytidine-5'-triphosphate, 5-propynyl-2'-deoxyuridine-5'-triphosphate, 6-azacytidine-5'-triphosphate, 6-azauridine-5'-triphosphate, 6-chloropurine riboside-5'-triphosphate, 7-deazaadenosine-5' -triphosphate, 7-deazaguanosine-5'-triphosphate, 8-azaadenosine-5'-triphosphate, 8-azidoadenosine-5'-triphosphate, benzimidazole-riboside-5'-triphosphate, N1-methyladenosine-5'-triphosphate, N1-methylguanosine-5'-triphosphate, N6-methyladenosine-5'-triphosphate, O6-methylguanosine-5'-triphosphate, pseudouridine-5'-triphosphate, or puromycin-5'-triphosphate,Preferably selected from xanthosine-5'-triphosphate, 5-methylcytidine-5'-triphosphate, 7-deazaguanosine-5'-triphosphate, 5-bromocytidine-5'-triphosphate, and pseudouridine-5'-triphosphate, pyridin-4-one ribonucleoside, 5-aza-uridine, 2-thio-5-aza-uridine, 2-thiouridine, 4-thio-pseudouridine, 2-thio-pseudouridine, 5-hydroxyuridine, 3-methyluridine, 5-carboxymethyl-uridine, 1-carboxymethyl-pseudouridine, 5-propynyl-uridine, 1-propynyl-pseudouridine, 5-taurinomethyluridine, 1-taurinomethyl-pseudouridine, 5-taurinomethyl-2-thio-uridine, 1-taurinomethyl-4-thio-uridine, 5-methyl-uridine, 1-methyl-pseudouridine, 4-thio-1-methyl-pseudouridine, 2-thio-1-methyl-pseudouridine, 1-methyl-1-deaza-pseudouridine, 2-thio-1-methyl-1-deaza-pseudouridine, dihydrouridine, dihydropseudouridine, 2-thio-dihydrouridine, 2- Thio-dihydropseudouridine, 2-methoxyuridine, 2-methoxy-4-thio-uridine, 4-methoxy-pseudouridine, and 4-methoxy-2-thio-pseudouridine, 5-aza-cytidine, pseudoisocytidine, 3-methyl-cytidine, N4-acetylcytidine, 5-formylcytidine, N4-methylcytidine, 5-hydroxymethylcytidine, 1-methyl-pseudoisocytidine, pyrrolo-cytidine, pyrrolo-pseudoisocytidine, 2-thio-cytidine, 2-thio-5-methyl-cytidine, 4-thio-pseudoisocytidine Socytidine, 4-thio-1-methyl-pseudoisocytidine, 4-thio-1-methyl-1-deaza-pseudoisocytidine, 1-methyl-1-deaza-pseudoisocytidine, zebularine, 5-aza-zebularine, 5-methyl-zebularine, 5-aza-2-thio-zebularine, 2-thio-zebularine, 2-methoxy-cytidine, 2-methoxy-5-methyl-cytidine, 4-methoxy-pseudoisocytidine, and 4-methoxy-1-methyl-pseudoisocytidine, 2-aminopurine, 2,6-diaminopurine, 7-deaza-adenine,7-Deaza-8-aza-adenine, 7-Deaza-2-aminopurine, 7-Deaza-8-aza-2-aminopurine, 7-Deaza-2,6-diaminopurine, 7-Deaza-8-aza-2,6-diaminopurine, 1-Methyladenosine, N6-Methyladenosine, N6-Isopentenyladenosine, N6-(cis-Hydroxyisopentenyl)adenosine, 2-Methylthio-N6-(cis-Hydroxyisopentenyl)adenosine, N6-Glycinylcarbamoyladenosine, N6-Threonylcarbamoyladenosine, 2-Methylthio-N6-threonylcarbamoyladenosine Bamoyl adenosine, N6,N6-dimethyl adenosine, 7-methyladenine, 2-methylthio-adenine, and 2-methoxy-adenine, inosine, 1-methyl-inosine, wyosine, wybutosine, 7-deaza-guanosine, 7-deaza-8-aza-guanosine, 6-thio-guanosine, 6-thio-7-deaza-8-aza-guanosine, 6-thio-7-deaza-8-aza-guanosine, 7-methyl-guanosine, 6-thio-7-methyl-guanosine, 7-methylinosine, 6-methoxy-guanosine, 1-methyl Guanosine, N2-methylguanosine, N2,N2-dimethylguanosine, 8-oxo-guanosine, 7-methyl-8-oxo-guanosine, 1-methyl-6-thio-guanosine, N2-methyl-6-thio-guanosine, and N2,N2-dimethyl-6-thioguanosine, 5'-O-(1-thiophosphate)-adenosine, 5'-O-(1-thiophosphate)-cytidine, 5'-O-(1-thiophosphate)-guanosine, 5'-O-(1-thiophosphate)-uridine, 5'-O-(1-thiophosphate)-pseudouridine, 6-azabicyclo[4.2.1]cinnamate, 5'-O-(1-thiophosphate)-cytidine, 5'-O-(1-thiophosphate)-guanosine ... thymidine, 2-thio-cytidine, alpha-thio-cytidine, pseudo-iso-cytidine, 5-aminoallyl-uridine, 5-iodo-uridine, N1-methyl-pseudouridine, 5,6-dihydrouridine, alpha-thio-uridine, 4-thio-uridine, 6-aza-uridine, 5-hydroxy-uridine, deoxy-thymidine, 5-methyl-uridine, pyrrolo-cytidine, inosine, alpha-thio-guanosine, 6-methyl-guanosine, 5-methyl-cytidine, 8-oxo-guanosine, 7-deaza-guanosine, N1-methyl-adenosine,Nucleotides for base modification selected from the group consisting of 2-amino-6-chloro-purine, N6-methyl-2-amino-purine, pseudo-iso-cytidine, 6-chloro-purine, N6-methyl-adenosine, alpha-thio-adenosine, 8-azido-adenosine, 7-deaza-adenosine are particularly preferred. Particularly preferred and suitable in the context of the present invention are pseudouridine (Ψ), N1-methylpseudouridine (m1Ψ), 5-methylcytosine, and 5-methoxyuridine. Thus, the artificial RNA defined herein can comprise at least one modified nucleotide selected from pseudouridine (Ψ), N1-methylpseudouridine (m1Ψ), 5-methylcytosine, and 5-methoxyuridine.

[0195] In a preferred embodiment, the artificial RNA of the present invention comprises at least one coding sequence, and at least one coding sequence is a pseudouridine (Ψ)-modified coding sequence.

[0196] Thus, in a preferred embodiment, an artificial RNA of the invention, or at least one coding sequence, comprises a nucleic acid sequence in which at least one or more than one, preferably all, uracil nucleotides are substituted with pseudouridine (Ψ) nucleotides.

[0197] In a further preferred embodiment, the artificial RNA of the present invention comprises at least one coding sequence, wherein at least one coding sequence is an N1-methylpseudouridine (m1Ψ) modified coding sequence.

[0198] Thus, in a preferred embodiment, the artificial RNA of the invention, or at least one coding sequence, comprises a nucleic acid sequence in which at least one or more, preferably all, uracil nucleotides are substituted with N1-methylpseudouridine (m1Ψ) nucleotides.

[0199] In a preferred embodiment, the artificial RNA of the present invention comprises at least one coding sequence, and at least one coding sequence is a codon-modified coding sequence.

[0200] In a preferred embodiment, at least one coding sequence of the present invention is a codon-modified coding sequence, and the amino acid sequence encoded by the at least one codon-modified coding sequence is preferably unaltered compared to the amino acid sequence encoded by the corresponding wild-type coding sequence.

[0201] The term "codon-modified coding sequence" relates to a coding sequence which differs in at least one codon (a triplet of nucleotides encoding one amino acid) compared to the corresponding wild-type coding sequence. Suitably, a codon-modified coding sequence in the context of the present invention may exhibit improved resistance to in vivo degradation, and / or improved stability in vivo, and / or improved translatability in vivo. Codon modification in the broadest sense exploits the degeneracy of the genetic code, which allows multiple codons to encode the same amino acid and can be used interchangeably (see Table 2) to optimize / modify coding sequences for in vivo applications as outlined above.

[0202] In particularly preferred embodiments of the first aspect, at least one sequence is a codon-modified coding sequence, wherein the codon-modified coding sequence is selected from a C-maximized coding sequence, a CAI-maximized coding sequence, a human codon usage adapted coding sequence, a G / C content modified coding sequence, and a G / C optimized coding sequence, or any combination thereof.

[0203] According to a preferred embodiment, the artificial RNA of the present invention can be modified to increase, preferably maximize, the C content of at least one coding sequence relative to the C content of the corresponding wild-type coding sequence (herein referred to as a "C-maximized coding sequence"). The amino acid sequence encoded by the C-maximized coding sequence of the RNA is preferably unchanged compared to the amino acid sequence encoded by the respective wild-type nucleic acid coding sequence. The generation of C-maximized nucleic acid sequences can be suitably carried out using the modification method according to WO 2015 / 062738. In this context, the disclosure of WO 2015 / 062738 is incorporated herein by reference. In the sequence listing, <223> Throughout the present disclosure, including the identifiers, the C-maximized coding sequence of a preferred RSV nucleic acid sequence is referred to by the abbreviation "opt2."

[0204] According to some embodiments, the artificial RNA of the present invention can be modified to alter the G / C content of at least one coding sequence of the present invention compared to the corresponding wild-type coding sequence (herein referred to as a "G / C content-modified coding sequence"). In this context, the terms "G / C optimized" or "G / C content-modified" refer to nucleic acids of the present invention, preferably artificial nucleic acids, that contain an altered, preferably increased, number of guanosine and / or cytosine nucleotides compared to the corresponding wild-type nucleic acid sequence. Such an increased number can be generated by substituting codons containing adenosine or thymidine nucleotides with codons containing guanosine or cytosine nucleotides. When generating an enriched G / C content in a DNA or RNA coding sequence, the degeneracy of the genetic code is exploited. In particular, in the case of RNA, sequences with an increased G (guanosine) / C (cytosine) content are more stable than sequences with an increased A (adenosine) / U (uracil) content. Preferably, the amino acid sequence encoded by a G / C content-modified coding sequence of a nucleic acid sequence remains unchanged compared to the amino acid sequence encoded by the respective wild-type nucleic acid coding sequence. Preferably, the G / C content of the coding sequence of an artificial nucleic acid sequence of the invention, e.g., an RNA sequence, is increased by at least 10%, preferably at least 20%, more preferably at least 30%, and most preferably at least 40%, compared to the G / C content of the coding sequence of the corresponding wild-type nucleic acid sequence (e.g., an RNA sequence) encoding a RSV antigen or fragment or variant thereof as defined herein.

[0205] According to a preferred embodiment, the artificial RNA of the present invention can be modified to alter the G / C content of at least one coding sequence of the present invention compared to the G / C content of the corresponding wild-type coding sequence (herein referred to as a "G / C content-optimized coding sequence"). "Optimized" in this context refers to a coding sequence whose G / C content has been increased, preferably to the maximum G / C content possible. The amino acid sequence encoded by a G / C content-optimized coding sequence of a nucleic acid sequence is preferably unchanged compared to the amino acid sequence encoded by the respective wild-type nucleic acid coding sequence. The generation of the above-mentioned G / C content-optimized RNA sequence can be suitably carried out using the G / C content optimization method described in WO 2002 / 098443. In this context, the entire scope of the disclosure of WO 2002 / 098443 is included in the present invention. <223> Throughout the present disclosure, including identifiers, the G / C content-optimized coding sequences of preferred RSV nucleic acid sequences are designated by the abbreviations "opt1, opt5, opt6, opt11."

[0206] According to some embodiments, the artificial RNA of the present invention can be modified to adapt the codons of at least one coding sequence of the present invention to human codon usage (herein referred to as a "human codon usage-adapted coding sequence"). Codons encoding the same amino acid occur at different frequencies in subjects, e.g., humans. Therefore, the coding sequence of the artificial RNA is preferably modified so that the frequencies of codons encoding the same amino acid correspond to the natural frequency of that codon according to human codon usage (see, e.g., Table 2). For example, for the amino acid alanine (Ala), the wild-type coding sequence is preferably adapted so that the codon "GCC" is used at a frequency of 0.40, the codon "GCT" is used at a frequency of 0.28, the codon "GCA" is used at a frequency of 0.22, the codon "GCG" is used at a frequency of 0.10 (see Table 2), etc. Thus, by applying such a procedure (exemplified by Ala) to the coding sequence of the artificial nucleic acid of the present invention, a sequence adapted to human codon usage is obtained. <223> Throughout the present disclosure, including identifiers, the human codon usage adapted coding sequence of the preferred RSV nucleic acid sequence is referred to by the abbreviation "opt3."

[0207] [Table 2]

[0208] According to some embodiments, the artificial RNA of the present invention can be modified to increase, or preferably maximize, the codon adaptation index (CAI) of at least one coding sequence of the present invention (referred to herein as a "CAI-maximized coding sequence"). Thus, for example, all codons of a wild-type nucleic acid sequence that are relatively rare in human cells are preferably replaced with respective codons that are frequent in human cells, where the frequent codons encode the same amino acid as the relatively rare codon. Preferably, the most frequent codon is used for each encoded amino acid (see Table 2, where the most frequent codon is indicated by an asterisk). Preferably, the artificial RNA of the present invention comprises at least one coding sequence, and the codon adaptation index (CAI) of at least one coding sequence is at least 0.5, at least 0.8, at least 0.9, or at least 0.95. Most preferably, the codon adaptation index (CAI) of at least one coding sequence is 1. For example, for the amino acid Ala, the wild-type coding sequence is adapted so that the most frequent human codon "GCC" is always used for said amino acid. Thus, such a procedure (exemplified by Ala) is applied to each amino acid coded by the coding sequence of the artificial RNA of the present invention to obtain a CAI-maximized coding sequence. <223> Throughout this disclosure, including the identifiers, the CAI-maximized coding sequence of the preferred RSV nucleic acid sequence is referred to by the abbreviation "opt4."

[0209] Therefore, in a particularly preferred embodiment, the artificial RNA of the first aspect is selected from the group consisting of SEQ ID NOs: 70 to 77, 485 to 492, 899 to 906, 1268 to 1275, 1637 to 1644, 2006 to 2013, 2375 to 2382, 2744 to 2751, 3113 to 3120, 3482 to 3489, 3851 to 3858, 4220 to 4227, 4589 to 4596, 4958 to 4965, 5327 to 5328, 5329 to 5330, 5331 to 5332, 5333 to 5334, 5335 to 5336, 5337 to 5338, 5339 to 5339 ... 334, 5696~5703, 6065~6072, 6434~6441, 6803~6810, 7172~7179, 7541~7548, 7910~7917, 11728~11734, 12097~12103, 12465~12472, 12834~12841, 13941~13948, 14310~14317, 14679~14686, 15048~15055, 15417~15418 424, 15786~15793, 13203~13210, 13572~13579, 16155~16162, 16524~16531, 16893~16900, 17262~17269, 17631~17638, 18000~18007, 18369~18376, 18738~18745, 19107~19114, 19476~19483, 21363~21384, 21389 9, 96%, 97%, 98%, or 99% identical to a codon-modified nucleic acid sequence selected from the group consisting of: 1-21410, 2-21411, 2-21412, 2-21413, 2-21414, 2-21415, 2-21416, 2-21417, 2-21418, 2-21419, 2-21420, 2-21421, 2-21422, 2-21423, 2-21424, 2-21425, 2-21426, 2-21427, 2-21428, 2-21429, 2-21430, 2-21431, 2-21432, 2-21433, 2-21434, 2-21435, 2-21436, 2-21437, 2-21438, 2-21439, 2-21440, 2-21441, 2-21442, 2-21443, 2-21444, 2-21445, 2-21446, 2-21447, 2-21448, 2-21449, 2-21450, 2-21451, 2-21452, 2-21453, 2-21454, 2-21455, 2-21456, 2-21457, 2-21458, 2-21459, 2-21460, 2-21461, 2-21 <223> This can also be obtained from the details given below.

[0210] In a particularly preferred embodiment, the artificial RNA of the first aspect is selected from the group consisting of SEQ ID NOs: 70 to 71, 75 to 77, 485 to 486, 490 to 492, 899 to 900, 904 to 906, 1268 to 1269, 1273 to 1275, 1637 to 1638, 1642 to 1644, 2006 to 2007, 2011 to 2013, 2375 to 2376, 2380 to 2382, 2744 to 2745, 2749 to 2751, 3113 to 3114, 3118 to 3120, 3482 to 3483, 3487 to 3489, 3852, 4221, 4590, 4959, 5328, 5697, 606 6, 6435, 6804, 7173, 7542, 7911, 3856~3858, 4225~4227, 4594~4596, 4963~4965, 5332~5334, 5701~5703, 6070~6072, 6439~6441, 6808~6810, 7177~717 9, 7546~7548, 7915~7917, 11728, 11732~11734, 12097, 12101~12103, 12465, 12466, 12470~12472, 12834, 12835, 12839~12841, 13941, 13942, 13946~1 3948, 14310, 14311, 14315~14317, 14679, 14680, 14684~14686, 15048, 15049, 15053~15055, 15417, 15418, 15422~15424, 15786, 15787, 15791~15793, 13203, 13204, 13208~13210, 13572, 13573, 13577~13579, 16155, 16156, 16160~16162, 16524, 16525, 16529~16531, 16893, 16894, 16898~16900, 17262 , 17263, 17267-17269, 17631, 17632, 17636-17638, 18000, 18001, 18005-18007, 18369, 18370, 18374-18376, 18738, 18739, 18743-18745, 19107, 19108, 19112-19114, 19476, 19477, 19481-19483, 21363-21384, 21389-21410, or fragments or variants of any of these sequences (see also Tables 3 and 4;opt1, 5, 6, 11) or at least 50%, 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical. Further information regarding each of these preferred nucleic acid sequences can be found in the Sequence Listing, in particular, under the identifiers <223> It can also be obtained from the details given below;

[0211] In a preferred embodiment, the artificial RNA of the present invention is selected from the group consisting of SEQ ID NOs: 73, 488, 902, 1271, 1640, 2009, 2378, 2747, 3116, 3485, 3854, 4223, 4592, 4961, 5330, 5699, 6068, 6437, 6806, 7175, 7544, 7913, 11730, 12099, 12468, 12837, 13944, 14313, 14682, 15051, 15420, 15789, 13206, 13575, 16158, 16527, 16896, 1 19110, 19479, or a fragment or variant of any of these sequences (see also Tables 3 and 4; opt3). Further information regarding each of these preferred nucleic acid sequences can be found in the Sequence Listing, in particular the identifiers <223> This can also be obtained from the details given below.

[0212] In a particularly preferred embodiment, the artificial RNA of the first aspect is selected from the group consisting of SEQ ID NOs: 72, 487, 901, 1270, 1639, 2008, 2377, 2746, 3115, 3484, 3853, 4222, 4591, 4960, 5329, 5698, 6067, 6436, 6805, 7174, 7543, 7912, 11729, 12098, 12467, 12836, 13943, 14312, 14681, 15050, 15419, 15788, 13205, 13574, 16157, 16526, 16 895, 17264, 17633, 18002, 18371, 18740, 19109, 19478, or a fragment or variant of any of these sequences (see also Tables 3 and 4; opt2). Further information regarding each of these preferred nucleic acid sequences can be found in the Sequence Listing, in particular the identifiers <223> This can also be obtained from the details given below.

[0213] In a preferred embodiment, the artificial RNA of the present invention is selected from the group consisting of SEQ ID NOs: 74, 489, 903, 1272, 1641, 2010, 2379, 2748, 3117, 3486, 3855, 4224, 4593, 4962, 5331, 5700, 6069, 6438, 6807, 7176, 7545, 7914, 11731, 12100, 12469, 12838, 13945, 14314, 14683, 15052, 15421, 15790, 13207, 13576, 16159, 16528, 16897 , 17266, 17635, 18004, 18373, 18742, 19111, 19480, or a fragment or variant of any of these sequences (see also Tables 3 and 4; opt4). Further information regarding each of these preferred nucleic acid sequences can be found in the Sequence Listing, in particular the identifiers <223> This can also be obtained from the details given below.

[0214] In some embodiments, the A / U content of the ribosome binding site environment of an artificial nucleic acid, particularly an artificial RNA of the present invention, can be increased compared to the A / U content of the ribosome binding site environment of its respective wild-type nucleic acid. This modification (increased A / U content around the ribosome binding site) increases the efficiency of ribosome binding to nucleic acids, preferably RNA. Effective binding of ribosomes to the ribosome binding site in turn has the effect of efficient translation of RNA. Thus, in particularly preferred embodiments, the artificial nucleic acids of the present invention comprise a ribosome binding site, also known as a "Kozak sequence," that is identical or at least 80%, 85%, 90%, or 95% identical to any of the sequences set forth in SEQ ID NOs: 41 and 42, or fragments or variants thereof.

[0215] Preferred RSV polypeptide and nucleic acid coding sequences ("cds") are provided in Tables 3A and 3B and Tables 4A and 4B.

[0216] Columns A to J of Tables 3A and 3B represent specific and preferred constructs of the present invention derived from RSV fusion (F) proteins, where column A provides a suitable sequence for F0, column B provides a suitable sequence for F-del, column C provides a suitable sequence for F0_DSCav1, column D provides a suitable sequence for F-del_DSCav1, column E provides a suitable sequence for F_DSCav1_mut1, column F provides a suitable sequence for F-del_DSCav1_mut1, column G provides a suitable sequence for F_DSCav1_mut2, column H provides a suitable sequence for F-del_DSCav1_mut2, column I provides a suitable sequence for F_DSCav1_mut3, and column J provides a suitable sequence for F-del_DSCav1_mut3. The sequence numbers of specific proteins provided in the sequence listing are given in row 2 ("PRT"). The SEQ ID NO of the corresponding wild-type / unmodified coding sequence is given in line 3 ("wt"). The SEQ ID NO of the codon-modified coding sequence corresponding to each protein construct is given in lines 4-10 ("optl", "opt2", "opt3", "opt4", "opt5", "opt6", "optll"). In Table 3A, the coding sequences derived from HRSV(A2) are given, and in Table 3B, the coding sequences derived from HRSV(Memphis-37) are given. Further information can be found in the sequence listing for each SEQ ID NO. <223> Given to the identifier.

[0217] [Table 3A]

[0218] [Table 3B]

[0219] Columns K to V of Tables 4A and 4B represent specific preferred constructs of the invention derived from RSV fusion (F) proteins, where column K provides a preferred sequence for F_DSCav1_mut0, column L provides a preferred sequence for F-del_DSCav1_mut0, column M provides a preferred sequence for F_DSCav1_mut4, column N provides a preferred sequence for F-del_DSCav1_mut4, column O provides a preferred sequence for F_DSCav1_mut5, and column P provides a preferred sequence for F_DSCav1_mut4. Column Q provides a suitable sequence for F_del_DSCav1_mut5, column Q provides a suitable sequence for F_del_DSCav1_mut6, column R provides a suitable sequence for F-del_DSCav1_mut6, column S provides a suitable sequence for F_DSCav1_mut7, column T provides a suitable sequence for F-del_DSCav1_mut7, column U provides a suitable sequence for F_DSCav1_mut8, and column V provides a suitable sequence for F-del_DSCav1_mut8. The SEQ ID NOs for specific proteins provided in the Sequence Listing are given in row 2 ("PRT"). The SEQ ID NOs for the corresponding codon-modified coding sequences for each protein construct are given in rows 3-9 ("opt1", "opt2", "opt3", "opt4", "opt5", "opt6", "opt11"). In Table 4A, coding sequences derived from HRSV(A2) are given, and in Table 4B, coding sequences derived from HRSV(Memphis-37) are given. Further information can be found in the SEQ ID NOs for each SEQ ID NO in the Sequence Listing. <223> Given to the identifier.

[0220] [Table 4A]

[0221] [Table 4B]

[0222] In some embodiments, the artificial RNA of the first aspect is monocistronic, bicistronic, or multicistronic.

[0223] In a preferred embodiment, the artificial RNA of the present invention is monocistronic.

[0224] The term "monocistronic nucleic acid" will be recognized and understood by those of skill in the art and is intended to mean, for example, an artificial RNA that contains only one coding sequence as defined herein. The terms "bicistronic nucleic acid, multicistronic nucleic acid" or "monocistronic RNA" as used herein will be recognized and understood by those of skill in the art and is intended to mean, for example, an artificial RNA that may have two (bicistronic) or even more (multicistronic) coding sequences.

[0225] In some embodiments, the artificial RNA of the present invention is monocistronic, and the coding sequence of the monocistronic artificial RNA encodes at least two different antigenic peptides or proteins, or fragments or variants thereof, derived from RSV F as defined herein. Thus, at least one coding sequence of the monocistronic artificial RNA can encode at least 2, 3, 4, 5, 6, 7, 8, or more antigenic peptides or proteins derived from RSV, preferably RSV F as defined herein, linked or not to an amino acid linker sequence, wherein the linker sequence can comprise a rigid linker, a flexible linker, a cleavable linker (e.g., a self-cleaving peptide), or a combination thereof (referred to herein as a "multi-antigen construct / nucleic acid").

[0226] In some embodiments, the artificial RNA of the present invention is bicistronic or multicistronic and comprises at least two coding sequences, wherein the at least two coding sequences encode two or more different antigenic peptides or proteins derived from RSV, preferably RSV F as defined herein, or any fragment or variant thereof. Thus, the coding sequences in the bicistronic or multicistronic artificial RNA preferably encode different antigenic proteins or peptides as defined herein, or fragments or variants thereof. Preferably, the coding sequences in the bicistronic or multicistronic artificial RNA are separated by at least one IRES (internal ribosome entry site) sequence. Thus, the term "encoding two or more antigenic peptides or proteins" is not intended to be limiting, and may refer to, for example, at least two, three, four, five, six, or more (preferably different) antigenic peptides or proteins of different RSVs, or fragments or variants thereof within the definitions set forth herein. Alternatively, the bicistronic or multicistronic artificial RNA may encode, for example, at least two, three, four, five, six, or more (preferably different) antigenic peptides or proteins from the same RSV, or fragments or variants within the scope of the definitions described herein. In this context, suitable IRES sequences can be selected from the list of nucleic acid sequences set forth in SEQ ID NOs: 1566-1662 of WO 2017 / 081082, or fragments or variants of these sequences. In this context, the disclosure of WO 2017 / 081082 regarding IRES sequences is incorporated herein by reference.

[0227] In the context of the present invention, it should be understood that specific combinations of coding sequences can be generated by any combination of monocistronic, bicistronic, and multicistronic artificial nucleic acids and / or multi-antigen constructs / nucleic acids to obtain nucleic acid compositions encoding multiple antigenic peptides or proteins as defined herein.

[0228] Preferably, an artificial RNA comprising at least one coding sequence as defined herein typically comprises a length of about 50 to about 20,000, or 500 to about 20,000 nucleotides, or about 500 to about 20,000 nucleotides, or about 500 to about 10,000 nucleotides, or about 1,000 to about 10,000 nucleotides, or preferably about 1,000 to about 5,000 nucleotides, or even more preferably about 1,000 to about 2,500 nucleotides.

[0229] According to a preferred embodiment, the artificial RNA of the first aspect can be an mRNA, a self-replicating RNA, a circular RNA, or a replicon RNA.

[0230] In some embodiments, the artificial RNA is a circular RNA. As used herein, "circular RNA" or "circRNA" should be understood as a circular polynucleotide capable of encoding at least one antigenic peptide or protein as defined herein. Thus, in a preferred embodiment, the circular RNA comprises at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV, or a fragment or variant thereof as defined herein. CircRNAs can be produced using various methods provided in the art. For example, U.S. Patent No. 6,210,931 teaches a method for synthesizing circRNAs by inserting a DNA fragment into a plasmid containing a sequence capable of spontaneous cleavage and self-cyclization. U.S. Patent No. 5,773,244 teaches the production of circRNAs by creating a DNA construct encoding an RNA cyclase ribozyme, expressing the DNA construct as RNA, and then self-splicing the RNA to generate intron-free circRNAs in vitro. WO 1992 / 001813 teaches the process of synthesizing linear polynucleotides, combining the linear nucleotides with complementary linking oligonucleotides under hybridization conditions, and ligating the linear polynucleotides to produce single-stranded circular nucleic acids.Those skilled in the art can also use the method provided in WO 2015 / 034925 or WO 2016 / 011222 to produce circular RNA.Therefore, the methods of producing circular RNA provided in US Patent No. 6,210,931, US Patent No. 5,773,244, WO 1992 / 001813, WO 2015 / 034925, and WO 2016 / 011222 are incorporated herein by reference.

[0231] In some embodiments, the artificial RNA is a replicon RNA. The term "replicon RNA" is recognized and understood by those skilled in the art and is intended to refer, for example, to an optimized, self-replicating artificial RNA construct. Such a construct comprises a replication element (replicase) derived from an alphavirus and the replacement of a structural viral protein with an artificial nucleic acid of interest (in the context of the present invention, an artificial nucleic acid comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV). Alternatively, the replicase can be provided in a separate construct comprising a replicase RNA sequence derived from, for example, Semliki Forest virus (SFV), Sindbis virus (SIN), Venezuelan equine encephalitis virus (VEE), Ross River virus (RRV), or other viruses belonging to the Alphaviridae family. Downstream of the replicase is a subgenomic promoter that controls replication of the artificial nucleic acid of the present invention, i.e., an artificial nucleic acid comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV.

[0232] In a preferred embodiment, the artificial RNA of the first aspect is mRNA.

[0233] The terms "RNA" and "mRNA" are recognized and understood by those skilled in the art and are intended to refer, for example, to a ribonucleic acid molecule, i.e., a polymer composed of nucleotides. These nucleotides are typically adenosine monophosphate, uridine monophosphate, guanosine monophosphate, and cytidine monophosphate monomers linked together along a so-called backbone. The backbone is formed by a phosphodiester bond between a first sugar, i.e., ribose, and the phosphate moiety of a second, adjacent monomer. A particular series of monomers is called an RNA sequence. mRNA (messenger RNA) typically provides a nucleotide sequence that can be translated into the amino acid sequence of a specific peptide or protein.

[0234] The artificial RNA, preferably mRNA, of the present invention can be prepared using any method known in the art, including chemical synthesis, such as solid-phase RNA synthesis, and in vitro methods such as RNA in vitro transcription reactions.

[0235] In a preferred embodiment, the artificial RNA, preferably mRNA, is obtained by in vitro transcription of RNA.

[0236] Thus, the RNA of the present invention is preferably in vitro transcribed RNA.

[0237] The term "RNA in vitro transcription" or "in vitro transcription" refers to a process by which RNA is synthesized in a cell-free system (in vitro). RNA can be obtained by DNA-dependent in vitro transcription of a suitable DNA template, which can be a linearized plasmid DNA template or a PCR-amplified DNA template, according to the present invention. The promoter for controlling RNA in vitro transcription can be any promoter for any DNA-dependent RNA polymerase. Specific examples of DNA-dependent RNA polymerases are T7, T3, SP6, or Syn5 RNA polymerase. In a preferred embodiment of the present invention, the DNA template is linearized with a suitable restriction enzyme before it is subjected to in vitro transcription of RNA.

[0238] Reagents used in in vitro transcription of RNA typically include a DNA template (linearized plasmid DNA or PCR product) having a promoter sequence with high binding affinity for a respective RNA polymerase, such as a bacteriophage-encoded RNA polymerase (T7, T3, SP6, or Syn5); ribonucleotide triphosphates (NTPs) of four bases (adenine, cytosine, guanine, and uracil); optionally, a cap analog as defined herein (e.g., m7G(5')ppp(5')G (m7G, m7G(5')ppp(5')(2'OMeG)pG, or m7G(5')ppp(5')(2'OMeA)pG)); optionally, further modified nucleotides as defined herein; and a DNA-dependent RNA polymerase (e.g., T7, T3, SP6, or Syn5) capable of binding to the promoter sequence in the DNA template. RNA polymerase; optionally, a ribonuclease (RNase) inhibitor to inactivate potentially contaminating RNases; optionally, a pyrophosphatase to degrade pyrophosphate that may inhibit in vitro transcription of RNA; MgCl2 to provide Mg2+ ions as a cofactor for the polymerase; and a buffer (TRIS or HEPES) to maintain an appropriate pH value (which may also contain an antioxidant (e.g., DTT) and / or a polyamine such as spermidine at optimal concentrations; an example is the TRIS-citrate buffer system disclosed in WO 2017 / 109161).

[0239] In some embodiments, the nucleotide mixture used in the in vitro transcription of RNA can further comprise modified nucleotides as defined herein, in which context preferred modified nucleotides include pseudouridine (Ψ), N1-methylpseudouridine (m1Ψ), 5-methylcytosine, and / or 5-methoxyuridine.

[0240] In a preferred embodiment, the nucleotide mixture (i.e., the fraction of each nucleotide in the mixture) used in the in vitro transcription reaction of RNA can be optimized for a given RNA sequence, preferably as described in WO 2015 / 188933.

[0241] In embodiments where it is necessary to generate more than one different artificial RNA as defined herein, for example, where it is necessary to generate 2, 3, 4, 5, 6, 7, 8, 9, 10 or more different artificial RNAs (e.g., encoding different RSV F antigens, or a combination of, for example, RSV F and RSV G; see the second aspect), the procedures described in WO 2017 / 109134 can be suitably used.

[0242] In the context of RNA vaccine production, it is necessary to provide GMP-grade RNA. GMP-grade RNA can be produced using a manufacturing process approved by regulatory authorities. Therefore, in a particularly preferred embodiment, RNA production is carried out under current Good Manufacturing Practice (GMP), preferably in accordance with WO 2016 / 180430, with various quality control steps at the DNA and RNA levels. In a preferred embodiment, the RNA of the present invention is GMP-grade RNA, particularly GMP-grade mRNA.

[0243] The resulting RNA product is preferably purified using PureMessenger® (CureVac, Tübingen, Germany; RP-HPLC according to WO 2008 / 077592) and / or tangential flow filtration (described in WO 2016 / 193206).

[0244] In a further preferred embodiment, the RNA, particularly the purified RNA, is freeze-dried according to WO 2016 / 165831 or WO 2011 / 069586 to obtain the thermostable dried artificial RNA (powder) as defined herein. The RNA, particularly the purified RNA of the invention, can also be dried using spray drying or spray freeze drying according to WO 2016 / 184575 or WO 2016184576 to obtain the thermostable RNA (powder) as defined herein. Accordingly, the disclosures of WO 2017 / 109161, WO 2015 / 188933, WO 2016 / 180430, WO 2008 / 077592, WO 2016 / 193206, WO 2016 / 165831, WO 2011 / 069586, WO 2016 / 184575, and WO 2016 / 184576 with respect to the production and purification of RNA are hereby incorporated by reference.

[0245] Thus, in a preferred embodiment, the RNA is dry RNA, in particular dry mRNA.

[0246] The term "dried RNA" as used herein should be understood as RNA that has been freeze-dried, or spray-dried, or spray-freeze-dried as defined above, to obtain a temperature-stable dry RNA (powder).

[0247] In a preferred embodiment, the artificial RNA of the present invention is purified RNA, in particular purified mRNA.

[0248] As used herein, the terms "purified RNA" or "purified mRNA" refer to RNA that has a higher degree of purity than the starting material (e.g., in vitro transcribed RNA) after a specific purification step (e.g., HPLC, TFF, oligo d(T) purification, precipitation step). Typical impurities that are essentially absent from purified RNA include peptides or proteins (e.g., enzymes derived from DNA-dependent RNA in vitro transcription, e.g., RNA polymerases, RNases, pyrophosphatases, restriction endonucleases, DNases), spermidine, BSA, incomplete RNA sequences, RNA fragments (short double-stranded RNA fragments, incomplete sequences, etc.), free nucleotides (modified nucleotides, conventional NTPs, cap analogs), template DNA fragments, buffer components (HEPES, TRIS, MgCl), etc. Other possible impurities that may result from, for example, fermentation procedures include bacterial impurities (bioburden, bacterial DNA) or impurities derived from the purification procedure (e.g., organic solvents). Therefore, in this regard, it is desirable for the "degree of RNA purity" to be as close to 100% as possible. The degree of RNA purity is also desirable for the amount of full-length RNA transcripts to be as close to 100% as possible.Therefore, as used herein, "purified RNA" has a purity of 75%, 80%, 85%, particularly 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, and most preferably 99% or more.The degree of purity can be determined, for example, by analytical HPLC, and the percentage corresponds to the ratio between the area of ​​the peak of target RNA and the total area of ​​all peaks representing by-products.Alternatively, the degree of purity can be determined, for example, by analytical agarose gel electrophoresis or capillary gel electrophoresis.

[0249] "Dried RNA" as defined herein and "purified RNA" as defined herein or "GMP-grade mRNA" as defined herein may have superior stability properties (in vitro, in vivo) and improved efficiency (e.g., greater translation ability of mRNA in vivo) and are therefore particularly suitable in the context of the present invention. Furthermore, "dried RNA" as defined herein and "purified RNA" or "GMP-grade mRNA" as defined herein may be particularly suitable for medical uses as defined herein.

[0250] The artificial RNA can be suitably modified by the addition of a 5'-cap structure, which preferably stabilizes the nucleic acid, as described herein. Thus, in a preferred embodiment, the artificial RNA of the first aspect comprises a 5'-cap structure, preferably m7G, cap0 (e.g., m7G(5')ppp(5')G), cap1 (e.g., m7G(5')ppp(5')(2'OMeG) or m7G(5')ppp(5')(2'OMeA)), cap2, modified cap0, or modified cap1 structure (which are generated using cap analogs as defined below).

[0251] The term "5'-cap structure," as used herein, is recognized and understood by those of skill in the art and is intended to refer, for example, to a modified nucleotide (cap analog), particularly a guanine nucleotide, added to the 5' end of an RNA molecule, such as an mRNA molecule. Preferably, the 5'-cap is added using a 5'-5'-triphosphate linkage (also referred to as m7GpppN). Further examples of 5'-cap structures include glyceryl, inverted deoxyabasic residue (moiety), 4',5' methylene nucleotide, 1-(beta-D-erythrofuranosyl) nucleotide, 4'-thionucleotide, carbocyclic nucleotide, 1,5-anhydrohexitol nucleotide, L-nucleotide, alpha-nucleotide, modified base nucleotide, threo-pentofuranosyl nucleotide, acyclic 3',4'-seconucleotide, acyclic 3,4-dihydroxybutyl nucleotide, acyclic 3,5 dihydroxypentyl nucleotide, 3'-3'-inverted nucleotide moiety, 3'-3'-inverted abasic moiety, 3'-2'-inverted nucleotide moiety, 3'-2'-inverted abasic moiety, 1,4-butanediol phosphate, 3'-phosphoramidate, hexyl phosphate, aminohexyl phosphate, 3'-phosphate, 3'-phosphorothioate, phosphorodithioate, or a bridged or non-bridged methylphosphonate moiety.

[0252] Additional 5'-cap structures that may be suitable in the context of the present invention are cap1 (additional methylation of the ribose of the nucleotide adjacent to m7GpppN), cap2 (additional methylation of the ribose of the second nucleotide downstream of m7GpppN), cap3 (additional methylation of the ribose of the third nucleotide downstream of m7GpppN), cap4 (additional methylation of the ribose of the fourth nucleotide downstream of m7GpppN), ARCA (anti-reverse cap analog), modified ARCA (e.g., phosphothioate-modified ARCA), inosine, N1-methyl-guanosine, 2'-fluoro-guanosine, 7-deaza-guanosine, 8-oxo-guanosine, 2-amino-guanosine, LNA-guanosine, and 2-azido-guanosine.

[0253] The 5'-cap (cap0 or cap1) structure can also be formed during chemical RNA synthesis or, preferably, during RNA in vitro transcription using a cap analog (co-transcriptional capping).

[0254] The term "cap analog," as used herein, will be recognized and understood by those of skill in the art and is intended to mean a non-polymerizable dinucleotide having a cap functionality that, when incorporated, for example, at the 5' end of a nucleic acid molecule, facilitates translation or localization and / or prevents degradation of the nucleic acid molecule, particularly an RNA molecule. By non-polymerizable, we mean that the cap analog is incorporated only at the 5' end, since it does not have a 5' triphosphate and cannot be extended in the 3' direction by a template-dependent polymerase, particularly a template-dependent RNA polymerase. Examples of cap analogs include, but are not limited to, chemical structures selected from the group consisting of m7GpppG, m7GpppA, m7GpppC; unmethylated cap analogs (e.g., GpppG); dimethylated cap analogs (e.g., m2,7GpppG), trimethylated cap analogs (e.g., m2,2,7GpppG), dimethylated symmetric cap analogs (e.g., m7Gpppm7G), or anti-inverted cap analogs (e.g., ARCA; m7,2'OmeGpppG, m7,2'dGpppG, m7,3'OmeGpppG, m7,3'dGpppG, and their tetraphosphate derivatives). Further cap analogues have been previously described (WO 2008 / 016473, WO 2008 / 157688, WO 2009 / 149253, WO 2011 / 015347, and WO 2013 / 059475). Further suitable cap analogs in this context are described in WO 2017 / 066793, WO 2017 / 066781, WO 2017 / 066791, WO 2017 / 066789, WO 2017 / 053297, WO 2017 / 066782, WO 2018075827, and WO 2017 / 066797, the disclosures of which regarding cap analogs are incorporated herein by reference.

[0255] A 5'-cap structure can be suitably added co-transcriptionally in an RNA in vitro transcription reaction as defined herein using a cap analog as defined herein.

[0256] In a preferred embodiment, the modified cap1 structure is generated using the cap analogs disclosed in WO 2017 / 053297, WO 2017 / 066793, WO 2017 / 066781, WO 2017 / 066791, WO 2017 / 066789, WO 2017 / 066782, WO 2018075827, and WO 2017 / 066797. In particular, any cap structure derivable from the structures disclosed in claims 1 to 5 of WO 2017 / 053297 can be suitably used to generate the modified cap1 structure by co-transcription. Furthermore, any cap structure derivable from the structure defined in claim 1 or claim 21 of WO 2018075827 can be suitably used to generate the modified cap1 structure by co-transcription.

[0257] A preferred cap analog is the dinucleotide cap analog mG(5')ppp(5')G(mG) or 3'-O-Me-mG(5')ppp(5')G, which co-transcriptionally generate the cap0 structure. A particularly preferred cap analog is the trinucleotide cap analog mG(5')ppp(5')(2'OMeA)pG or mG(5')ppp(5')(2'OMeG)pG, which co-transcriptionally generate the cap1 structure.

[0258] In this context, the RNA of the present invention preferably comprises the Cap1 structure defined above, which preferably results in increased protein expression, such as by high capping efficiency and improved translation efficiency. More preferably, the RNA of the present invention comprising the Cap1 structure exhibits reduced stimulation of the innate immune system compared to a Cap0 construct of the same nucleic acid sequence. Those skilled in the art are aware of methods for determining translation efficiency, capping degree, and immune stimulation.

[0259] In a particularly preferred embodiment, the artificial RNA of the first aspect of the present invention comprises a cap1 structure, which can be formed enzymatically or co-transcriptionally (e.g., using m7G(5')ppp(5')(2'OMeA)pG or m7G(5')ppp(5')(2'OMeG)pG analogs).

[0260] In a preferred embodiment, the artificial RNA of the first aspect comprises a m7G(5')ppp(5')(2'OMeA)pG cap structure. In such an embodiment, the coding RNA comprises a 5'-terminal m7G cap and an additional methylation of the ribose of the adjacent nucleotide of m7GppN (in this case, a 2'O-methylated adenosine).

[0261] In another preferred embodiment, the artificial RNA of the first aspect comprises an m7G(5')ppp(5')(2'OMeG)pG cap structure. In such an embodiment, the coding RNA comprises an m7G cap at the 5' end and an additional methylation of the ribose of the adjacent nucleotide (in this case, a 2'O-methylated guanosine).

[0262] Therefore, whenever a suitable RNA or mRNA sequence is referred to in the context of the present invention, the first nucleotide of said RNA or mRNA sequence, i.e. the nucleotide downstream of the m7G(5')ppp structure, may be a 2'O-methylated guanosine or a 2'O-methylated adenosine.

[0263] Thus, in other embodiments, the artificial RNA of the invention can comprise a 5' cap sequence element according to SEQ ID NO: 43 or 21321, or a fragment or variant thereof.

[0264] In other embodiments, the 5'-cap structure is added via enzymatic capping using a capping enzyme (e.g., vaccinia virus capping enzyme, commercially available capping kits) to generate a cap0, cap1, or cap2 structure. In other embodiments, the 5'-cap structure (cap0, cap1) is added via enzymatic capping using an immobilized capping enzyme, e.g., using a capping reactor (WO 2016 / 193226).

[0265] In a preferred embodiment, the artificial RNA of the present invention comprises at least one poly(A) sequence, preferably comprising 30 to 150 adenosine nucleotides.

[0266] In a preferred embodiment, the poly(A) sequence, suitably located at the 3'-terminus, contains 10 to 500 adenosine nucleotides, 10 to 200 adenosine nucleotides, 40 to 200 adenosine nucleotides, or 40 to 150 adenosine nucleotides. In a particularly preferred embodiment, the poly(A) sequence contains about 64 adenosine nucleotides. In an even more particularly preferred embodiment, the poly(A) sequence contains about 75 adenosine nucleotides. In an even more particularly preferred embodiment, the poly(A) sequence contains about 100 adenosine nucleotides.

[0267] The terms "poly(A) sequence," "poly(A) tail," or "3'-poly(A) tail" as used herein are recognized and understood by those skilled in the art and are intended to refer to, for example, a series of adenosine nucleotides, typically up to about 1,000 adenosine nucleotides, located at the 3' end of an RNA. Preferably, the poly(A) sequence is essentially homopolymeric, e.g., a poly(A) sequence of essentially 100 nucleotides in length, such as 100 adenosine nucleotides. In other embodiments, the poly(A) sequence may be interrupted by at least one nucleotide that is different from adenosine nucleotides; for example, a poly(A) sequence of 100 adenosine nucleotides may have a length of more than 100 nucleotides (including 100 adenosine nucleotides and the at least one nucleotide that is different from adenosine nucleotides).

[0268] In the context of the present invention, poly(A) sequences may be located within an mRNA or any other nucleic acid molecule, such as, for example, within a DNA vector, e.g., within a vector that serves as a template for the production of RNA, preferably mRNA, by transcription of said DNA vector.

[0269] Preferably, the poly(A) sequence of the artificial RNA is obtained from a DNA template during in vitro transcription of the RNA. In another embodiment, the poly(A) sequence is obtained in vitro by common methods of chemical synthesis, not necessarily by transcription from a DNA template. In another embodiment, the poly(A) sequence is produced by enzymatic polyadenylation of the RNA (after in vitro transcription of the RNA) using a commercially available polyadenylation kit and corresponding protocols known in the art, or alternatively, by using immobilized poly(A) polymerase in a polyadenylation reactor (described in WO 2016 / 174271).

[0270] In some embodiments, the artificial RNA of the present invention can include a poly(A) sequence derived from a vector and can include at least one additional poly(A) sequence generated by enzymatic polyadenylation (e.g., as described in WO 2016 / 091391).

[0271] In a preferred embodiment, the artificial RNA of the present invention contains at least one poly(C) sequence, preferably containing 10 to 40 cytosine nucleotides.

[0272] In a preferred embodiment, the poly(C) sequence, suitably located at the 3'-terminus, contains 10 to 200 cytosine nucleotides, 10 to 100 cytosine nucleotides, 20 to 70 cytosine nucleotides, 20 to 60 cytosine nucleotides, or 10 to 40 cytosine nucleotides. In a particularly preferred embodiment, the poly(C) sequence contains about 30 cytosine nucleotides.

[0273] The term "poly(C) sequence" as used herein will be recognized and understood by those skilled in the art and refers to, for example, a sequence of cytosine nucleotides, typically up to about 200 cytosine nucleotides, located at the 3' end of an RNA. In the context of the present invention, a poly(C) sequence may be located within an mRNA or any other nucleic acid molecule, such as, for example, a DNA vector, e.g., a vector that serves as a template for producing RNA, preferably mRNA, by transcription of the vector.

[0274] Preferably, the poly(C) sequences in the RNA sequences of the present invention are obtained by in vitro transcription of RNA from a DNA template. In other embodiments, the poly(C) sequences are obtained in vitro by common methods of chemical synthesis, without necessarily being transcribed from a DNA template.

[0275] In other embodiments, the artificial RNA of the present invention does not contain a poly(C) sequence as defined herein.

[0276] In a preferred embodiment, the artificial RNA of the first aspect comprises at least one histone stem loop.

[0277] The term "histone stem loop" as used herein is intended to refer to a nucleic acid sequence recognized and understood by those skilled in the art, e.g., found primarily in histone mRNAs. Exemplary histone stem loop sequences are described in Lopez et al. (Davila Lopez et al. (2008), RNA, 14(1)). The stem loop of a histone pre-mRNA is typically followed by a purine-rich sequence known as a histone downstream element (HDE). These pre-mRNAs are processed in the nucleus by a single endonucleolytic cleavage approximately 5 nucleotides downstream of the stem loop, catalyzed by U7 snRNP via base pairing between the U7 snRNA and the HDE.

[0278] The histone stem-loop sequence / structure can be suitably selected from the histone stem-loop sequences disclosed in WO 2012 / 019780, the disclosure of which is incorporated herein by reference. The histone stem-loop sequence that can be used in the present invention is preferably derived from formula (I) or (II) of WO 2012 / 019780. According to a further preferred embodiment, the RNA defined herein can comprise at least one histone stem-loop sequence derived from at least one of specific formulas (Ia) and (IIa) of WO 2012 / 019780.

[0279] In a particularly preferred embodiment, the artificial RNA of the present invention comprises at least one histone stem loop, said histone stem loop comprising a nucleic acid sequence according to SEQ ID NO: 39 or 40, or a fragment or variant thereof.

[0280] In other embodiments, the artificial RNA of the first aspect does not comprise a histone stem loop as defined herein.

[0281] In a further embodiment, the artificial RNA of the invention comprises a 3'-terminal sequence element, which should be understood as a sequence element comprising a poly(A) sequence and a histone stem-loop sequence, and which is located at the 3'-end of the artificial RNA of the invention.

[0282] In other embodiments, the artificial RNA of the present invention can include a 3'-terminal sequence element according to SEQ ID NOs: 44 to 63 or 21322 to 21328, or a fragment or variant thereof.

[0283] In a preferred embodiment, the artificial RNA of the present invention comprises at least one pseudouridine (Ψ) modified coding sequence.

[0284] Thus, in a preferred embodiment, an artificial RNA of the invention, or at least one coding sequence, comprises a nucleic acid sequence in which at least one or more than one, preferably all, uracil nucleotides are substituted with pseudouridine (Ψ) nucleotides.

[0285] In a further preferred embodiment, the artificial RNA of the present invention comprises at least one N1-methylpseudouridine (m1Ψ) modified coding sequence.

[0286] Thus, in a preferred embodiment, the artificial RNA of the invention, or at least one coding sequence, comprises a nucleic acid sequence in which at least one or more, preferably all, uracil nucleotides are substituted with N1-methylpseudouridine (m1Ψ) nucleotides.

[0287] In a preferred embodiment of the first aspect, the artificial RNA, preferably the mRNA, comprises, preferably in the 5' to 3' direction, the following elements a) to i): a) a 5'-cap structure, preferably a 5'-cap structure described herein; b) optionally, at least one 5'-UTR as described herein, preferably selected from SEQ ID NOs: 1 to 22; c) a ribosome binding site, preferably a ribosome binding site described herein; d) at least one coding sequence described herein, preferably at least one coding sequence described in Table 3 and Table 4; e) at least one 3'-UTR described herein, preferably selected from SEQ ID NOs: 23 to 38; f) optionally a poly(A) sequence, preferably a poly(A) sequence as described herein; g) optionally a poly(C) sequence, preferably a poly(C) sequence as described herein; h) optionally a histone stem loop, preferably a histone stem loop as described herein; i) optionally a 3'-terminal sequence element as described herein, preferably a 3'-terminal sequence element according to SEQ ID NOs: 44 to 63 or 21322 to 21328; and optionally, at least one or more, preferably all, uracil nucleotides are substituted with pseudouridine (Ψ) or N1-methylpseudouridine (m1Ψ) nucleotides.

[0288] In a further preferred embodiment of the first aspect, the artificial RNA, preferably the mRNA, comprises, preferably in the 5' to 3' direction, the following elements: a) a 5'-cap structure, preferably a 5'-cap structure described herein, most preferably a Cap1 structure; b) 3'-UTR and 5'-UTR elements according to a-1, a-4, c-1, e-4, g-2, i-2, or i-3 described herein; c) a ribosome binding site, preferably a ribosome binding site described herein; d) at least one coding sequence as described herein, located between the 5'-UTR and the 3'-UTR, preferably downstream of the 5'-UTR and upstream of the 3'-UTR, and preferably selected from any of those listed in Table 3 and Table 4; e) optionally a poly(A) sequence, preferably a poly(A) sequence as described herein; f) optionally a poly(C) sequence, preferably a poly(C) sequence as described herein; g) optionally a histone stem loop, preferably a histone stem loop as described herein; h) optionally a 3'-terminal sequence element as described herein, preferably a 3'-terminal sequence element according to SEQ ID NOs: 44 to 63, 21322 to 21328; and optionally, at least one or more, preferably all, uracil nucleotides are substituted with pseudouridine (Ψ) or N1-methylpseudouridine (m1Ψ) nucleotides.

[0289] In a further preferred embodiment of the first aspect, the artificial RNA, preferably the mRNA, comprises, preferably in the 5' to 3' direction, the following elements: a) a 5'-cap structure, preferably a 5'-cap structure described herein, most preferably a Cap1 structure; b) 3'-UTR and 5'-UTR elements according to a-1 or i-3 described herein; c) a ribosome binding site, preferably a ribosome binding site described herein; d) at least one coding sequence as described herein, located between the 5'-UTR and the 3'-UTR, preferably downstream of the 5'-UTR and upstream of the 3'-UTR, and preferably selected from any of those listed in Table 3 and Table 4; e) optionally a histone stem loop, preferably a histone stem loop as described herein; f) a poly(A) sequence, preferably a poly(A) sequence containing about 100 adenosine nucleotides; g) optionally a 3'-terminal sequence element as described herein, preferably a 3'-terminal sequence element according to SEQ ID NOs: 21322 to 21328; and optionally, at least one or more, preferably all, uracil nucleotides are substituted with pseudouridine (Ψ) or N1-methylpseudouridine (m1Ψ) nucleotides.

[0290] In a further preferred embodiment of the first aspect, the artificial RNA, preferably the mRNA, comprises in the 5' to 3' direction the following elements: a) a 5'-cap structure, preferably a 5'-cap structure described herein, most preferably a Cap1 structure; b) 3'-UTR and 5'-UTR elements according to a-1, a-4, c-1, e-4, g-2, i-2, or i-3 described herein; c) a ribosome binding site, preferably a ribosome binding site described herein; d) at least one coding sequence as described herein, located between the 5'-UTR and the 3'-UTR, preferably downstream of the 5'-UTR and upstream of the 3'-UTR, and preferably selected from the group consisting of SEQ ID NOs: 69-77, 484-492, 899-906, 1268-1275, 1637-1644, 2006-2013, 2375-2382, 274 4~2751, 3113~3120, 3482~3489, 3851~3858, 4220~4227, 4589~4596, 4958~4965, 5327~5334, 5696~5703, 6065~6072, 6434~6441, 6803~6810, 7172~7179, 7541~7548, 7910~7917, 11727~11734, 1 2096~12103, 12465~12472, 12834~12841, 13941~13948, 14310~14317, 14679~14686, 15048~15055, 15417~15424, 15786~15793, 13203~13210, 13572~13579, 16155~16162, 16524~16531, 1689 at least one coding sequence selected from any of: 3-16900, 17262-17269, 17631-17638, 18000-18007, 18369-18376, 18738-18745, 19107-19114, 19476-19483, 21363-21384, 21389-21410 (or a fragment or variant thereof); e) a poly(A) sequence containing approximately 64 adenosines; f) a poly(C) sequence containing approximately 30 cysteines; g) a histone stem loop according to SEQ ID NO: 39 or 40; and optionally, at least one or more, preferably all, uracil nucleotides are substituted with pseudouridine (Ψ) or N1-methylpseudouridine (m1Ψ) nucleotides.

[0291] In a further preferred embodiment of the first aspect, the artificial RNA, preferably the mRNA, comprises in the 5' to 3' direction the following elements: a) a 5'-cap structure, preferably a 5'-cap structure described herein, most preferably a Cap1 structure; b) 3'-UTR and 5'-UTR elements according to a-1, a-4, c-1, e-4, g-2, i-2, or i-3 described herein; c) a ribosome binding site, preferably a ribosome binding site described herein; d) at least one coding sequence as described herein, located between the 5'-UTR and the 3'-UTR, preferably downstream of the 5'-UTR and upstream of the 3'-UTR, and preferably selected from the group consisting of SEQ ID NOs: 69-77, 484-492, 899-906, 1268-1275, 1637-1644, 2006-2013, 2375-2382, 274 4~2751, 3113~3120, 3482~3489, 3851~3858, 4220~4227, 4589~4596, 4958~4965, 5327~5334, 5696~5703, 6065~6072, 6434~6441, 6803~6810, 7172~7179, 7541~7548, 7910~7917, 11727~11734, 1 2096~12103, 12465~12472, 12834~12841, 13941~13948, 14310~14317, 14679~14686, 15048~15055, 15417~15424, 15786~15793, 13203~13210, 13572~13579, 16155~16162, 16524~16531, 1689 at least one coding sequence selected from any of: 3-16900, 17262-17269, 17631-17638, 18000-18007, 18369-18376, 18738-18745, 19107-19114, 19476-19483, 21363-21384, 21389-21410 (or a fragment or variant thereof); e) a poly(A) sequence containing approximately 64 adenosines; f) a histone stem loop according to SEQ ID NO: 39 or 40; and optionally, at least one or more, preferably all, uracil nucleotides are substituted with pseudouridine (Ψ) or N1-methylpseudouridine (m1Ψ) nucleotides.

[0292] Preferred RSV polypeptide, nucleic acid, and mRNA sequences are shown in Tables 5A, 5B and Tables 6A, 6B.

[0293] In Tables 5A and 5B, the protein design is shown in row 1. In the table, columns A to J represent specific suitable constructs of the present invention derived from the RSV fusion (F) protein, where column A represents a suitable sequence for F0, column B represents a suitable sequence for F-del, column C represents a suitable sequence for F0_DSCav1, column D represents a suitable sequence for F-del_DSCav1, column E represents a suitable sequence for F_DSCav1_mut1, column F represents a suitable sequence for F-del_DSCav1_mut1, column G represents a suitable sequence for F_DSCav1_mut2, column H represents a suitable sequence for F-del_DSCav1_mut2, column I represents a suitable sequence for F_DSCav1_mut3, and column J represents a suitable sequence for F-del_DSCav1_mut3.

[0294] Protein designs are shown in row 1 (columns A-J), and the specific protein SEQ ID NOs as shown in the Sequence Listing are shown in row 2 ("Protein"). The SEQ ID NOs for the corresponding coding sequences for each protein construct are shown in row 3 ("cds", compare Table 3 for various cds optimizations). Further information regarding the type of codon-modified coding sequence (e.g., opt1, opt2, opt3, opt4, opt5, opt6, opt11) is provided in the table below each SEQ ID NO in the Sequence Listing. <223> The identifiers are listed in Table 3. SEQ ID NOs for the corresponding mRNA constructs containing the coding sequences and preferred 3'-UTRs and 5'-UTRs according to the present invention are listed in rows 4-47 (mRNA designs a-1 to i-3 described herein). Table 5A shows mRNA sequences derived from HRSV (A2), and Table 5B shows mRNA sequences derived from HRSV (Memphis-37). Further information regarding the type of coding sequence included in the mRNA construct (e.g., wt, opt1, opt2, opt3, opt4, opt5, opt6, opt11) can be found in the sequence listing for each SEQ ID NO. <223> It is written in the identifier.

[0295] [Table 5A-1] [Table 5A-2]

[0296] [Table 5B-1] [Table 5B-2]

[0297] In Tables 6A and 6B, the protein designs are shown in row 1. In the tables, columns K to V represent specific suitable constructs of the invention derived from the RSV fusion (F) protein, where column K represents a suitable sequence for F_DSCav1_mut0, column L represents a suitable sequence for F-del_DSCav1_mut0, column M represents a suitable sequence for F_DSCav1_mut4, column N represents a suitable sequence for F-del_DSCav1_mut4, column O represents a suitable sequence for F_DSCav1_mut5, and column P represents a suitable sequence for F-del_DSCav1_mut5. Column Q shows a suitable sequence for F_DSCav1_mut6, column R shows a suitable sequence for F-del_DSCav1_mut6, column S shows a suitable sequence for F_DSCav1_mut7, column T shows a suitable sequence for F-del_DSCav1_mut7, column U shows a suitable sequence for F_DSCav1_mut8, and column V shows a suitable sequence for F-del_DSCav1_mut8.

[0298] Protein designs are shown in the first row (columns K-V). Specific protein SEQ ID NOs shown in the Sequence Listing are listed under "PRT" in row 2. The SEQ ID NOs of the corresponding coding sequences for each protein construct are listed under "cds" in row 3 (compare Table 4 for various cds optimizations). Further information regarding the type of codon-modified coding sequence (e.g., opt1, opt2, opt3, opt4, opt5, opt6, opt11) is provided below each SEQ ID NO in the Sequence Listing. <223> The identifiers and sequences are listed in Table 4. The sequence numbers of the corresponding mRNA constructs containing the coding sequences and the preferred 3'-UTRs and 5'-UTRs of the present invention are listed in the following lines (mRNA designs "a-1" to "i-3" described herein). Table 6A shows mRNA sequences derived from HRSV(A2), and Table 6B shows mRNA sequences derived from HRSV(Memphis-37).

[0299] Further information regarding the type of coding sequence (e.g., wt, opt1, opt2, opt3, opt4, opt5, opt6, opt11) included in the mRNA construct is provided in the Sequence Listing for each SEQ ID NO. <223> It is written in the identifier.

[0300] Table 6A: Preferred mRNA constructs encoding RSV F from HRSV (A2) (rows K-V)

Table 6A-1

Table 6A-2

[0301] Table 6B: Preferred mRNA constructs encoding RSV F derived from HRSV (Memphis-37) (columns K-V)

Table 6B-1

Table 6B-2

[0302] In a preferred embodiment, the artificial RNA is selected from the group consisting of SEQ ID NOs: 78 to 482, 493 to 897, 907 to 1266, 1276 to 1635, 1645 to 2004, 2014 to 2373, 2383 to 2742, 2752 to 3111, 3121 to 3480, 3490 to 3849, 3859 to 4218, 4228 to 4587, 4597 to 4956, 4966 to 5325, 5335 to 5694, 5704 to 6063, 6073 to 6432, and 6442 to 680. 1, 6811~7170, 7180~7539, 7549~7908, 7918~8277, 8278, 21415~21480, 21561~21626, 11735~12094, 12104~12463, 12473~12832, 12842~13201, 13949~14308, 14318~14677, 14687~15046, 15056~15415, 15425~15784, 15794~16153, 13211~1 3570, 13580 to 13939, 16163 to 16522, 16532 to 16891, 16901 to 17260, 17270 to 17629, 17639 to 17998, 18008 to 18367, 18377 to 18736, 18746 to 19105, 19115 to 19474, 19484 to 19843, 21489 to 21552, and 21635 to 21698, or a fragment or variant of any of these sequences. The nucleic acid sequence of the present invention may comprise or consist of an RNA sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the nucleic acid sequence of the present invention, optionally in which at least one or more, and preferably all, uracil nucleotides are substituted by pseudouridine (Ψ) nucleotides or N1-methylpseudouridine (m1Ψ) nucleotides.

[0303] In a preferred embodiment, the artificial RNA comprises (a) at least one heterologous 5' untranslated region (5'-UTR) and / or at least one heterologous 3' untranslated region (3'-UTR); and (b) at least one coding sequence encoding at least one antigenic peptide or protein from RSV F, or a fragment or variant thereof, operably linked to the 3'-UTR and / or 5'-UTR. - said artificial RNA comprises or consists of an RNA sequence identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 78 to 482, 11735 to 12094, 21415 to 21417, 21561 to 21563, 21489, 21490, 21635, 21636 (encoding F0), or a fragment or variant of any of these sequences; - said artificial RNA comprises or consists of an RNA sequence identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 493 to 897, 12104 to 12463, 21418 to 21420, 21564 to 21566, 21491, 21492, 21637, 21638 (encoding F-del), or a fragment or variant of any of these sequences; - the artificial RNA comprises or consists of an RNA sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 907 to 1266, 12473 to 12832, 21421 to 21423, 21567 to 21569, 21493 to 21495, 21639 to 21641 (encoding F0_DSCav1), or a fragment or variant of any of these sequences; - the artificial RNA comprises or consists of an RNA sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1276-1635, 8278, 12842-13201, 21424-21426, 21570-21572, 21496-21498, 21642-21644 (encoding F-del_DSCav1), or a fragment or variant of any of these sequences; - said artificial RNA comprises or consists of an RNA sequence identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1645-2004, 13949-14308, 21433-21435, 21579-21581, 21505-21507, 21651-21653 (encoding F_DSCav1_mut1), or a fragment or variant of any of these sequences; - said artificial RNA comprises or consists of an RNA sequence identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 2014 to 2373, 14318 to 14677, 21436 to 21438, 21582 to 21584, 21508 to 21510, 21654 to 21656 (encoding F-del_DSCav1_mut1), or a fragment or variant of any of these sequences; - said artificial RNA comprises or consists of an RNA sequence identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 2383 to 2742, 14687 to 15046, 21439 to 21441, 21585 to 21587, 21511 to 21513, 21657 to 21659 (encoding F_DSCav1_mut2), or a fragment or variant of any of these sequences; - said artificial RNA comprises or consists of an RNA sequence identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 2752 to 3111, 15056 to 15415, 21442 to 21444, 21588 to 21590, 21514 to 21516, 21660 to 21662 (encoding F-del_DSCav1_mut2), or a fragment or variant of any of these sequences; - said artificial RNA comprises or consists of an RNA sequence identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 3121-3480, 15425-15784, 21445-21447, 21591-21593, 21517-21519, 21663-21665 (encoding F_DSCav1_mut3), or a fragment or variant of any of these sequences; - said artificial RNA comprises or consists of an RNA sequence identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 3490 to 3849, 15794 to 16153, 21448 to 21450, 21594 to 21596, 21520 to 21522, 21666 to 21668 (encoding F-del_DSCav1_mut3), or a fragment or variant of any of these sequences; - said artificial RNA comprises or consists of an RNA sequence identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 3859 to 4218, 13211 to 13570, 21427 to 21429, 21573 to 21575, 21499 to 21501, 21645 to 21647 (encoding F_DSCav1_mut0), or a fragment or variant of any of these sequences; - said artificial RNA comprises or consists of an RNA sequence identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 4228-4587, 13580-13939, 21430-21432, 21576-21578, 21502-21504, 21648-21650 (encoding F-del_DSCav1_mut0), or a fragment or variant of any of these sequences; - said artificial RNA comprises or consists of an RNA sequence identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 4597 to 4956, 16163 to 16522, 21451 to 21453, 21597 to 21599, 21523 to 21525, 21669 to 21671 (encoding F_DSCav1_mut4), or a fragment or variant of any of these sequences; - said artificial RNA comprises or consists of an RNA sequence identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 4966-5325, 16532-16891, 21454-21456, 21600-21602, 21526-21528, 21672-21674 (encoding F-del_DSCav1_mut4), or a fragment or variant of any of these sequences; - said artificial RNA comprises or consists of an RNA sequence identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 5335-5694, 16901-17260, 21457-21459, 21603-21605, 21529-21531, 21675-21677 (encoding F_DSCav1_mut5), or a fragment or variant of any of these sequences; - said artificial RNA comprises or consists of an RNA sequence identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 5704 to 6063, 17270 to 17629, 21460 to 21462, 21606 to 21608, 21532 to 21534, 21678 to 21680 (encoding F-del_DSCav1_mut5), or a fragment or variant of any of these sequences; - said artificial RNA comprises or consists of an RNA sequence identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 6073 to 6432, 17639 to 17998, 21463 to 21465, 21609 to 21611, 21535 to 21537, 21681 to 21683 (encoding F_DSCav1_mut6), or a fragment or variant of any of these sequences; - said artificial RNA comprises or consists of an RNA sequence identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 6442-6801, 18008-18367, 21466-21468, 21612-21614, 21538-21540, 21684-21686 (encoding F-del_DSCav1_mut6), or a fragment or variant of any of these sequences; - said artificial RNA comprises or consists of an RNA sequence identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 6811 to 7170, 18377 to 18736, 21469 to 21471, 21615 to 21617, 21541 to 21543, 21687 to 21689 (encoding F_DSCav1_mut7), or a fragment or variant of any of these sequences; - said artificial RNA comprises or consists of an RNA sequence identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 7180-7539, 18746-19105, 21472-21474, 21618-21620, 21544-21546, 21690-21692 (encoding F-del_DSCav1_mut7), or a fragment or variant of any of these sequences; - said artificial RNA comprises or consists of an RNA sequence identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 7549-7908, 19115-19474, 21475-21477, 21621-21623, 21547-21549, 21693-21695 (encoding F_DSCav1_mut8), or a fragment or variant of any of these sequences; - the artificial RNA comprises or consists of an RNA sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 7918-8277, 19484-19843, 21478-21480, 21624-21626, 21550-21552, 21696-21698 (encoding F-del_DSCav1_mut8), or a fragment or variant of any of these sequences, Optionally, at least one or more, preferably all, uracil nucleotides are substituted with pseudouridine (Ψ) or N1-methylpseudouridine (m1Ψ) nucleotides.

[0304] In a particularly preferred embodiment, the artificial RNA comprises (a) at least one heterologous 5' untranslated region (5'-UTR) and / or at least one heterologous 3' untranslated region (3'-UTR) and (b) a RSV gene operably linked to the 3'-UTR and / or 5'-UTR. The artificial RNA comprises at least one coding sequence encoding at least one antigenic peptide or protein derived from F, or a fragment or variant thereof, wherein the artificial RNA comprises or consists of an RNA sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 5704-6063, 17270-17629, 21460-21462, 21606-21608, 21532-21534, 21678-21680 (encoding F-del_DSCav1_mut5), or a fragment or variant of any of these sequences.

[0305] As explained throughout the specification, further information regarding suitable amino acid or nucleic acid sequences (coding sequences, mRNA sequences) can be found in the sequence listing, in particular in the identifiers 101 ... <223> It can also be obtained from the details given below.

[0306] For example, the numerical identifier of SEQ ID NO: 68 in the sequence listing <223> is as follows: "derived and / or modified protein sequence (artificial) from HRSV(A2)_F0". Throughout the sequence listing, numeric identifiers <223> The information given below has the same structure:<CONSTRUCT_IDENTIFIER> from<SEQUENCE_DESCRIPTOR> " should be considered to apply.<SEQUENCE_DESCRIPTOR> relates to the type of sequence (e.g., "derived and / or modified protein sequence," "derived and / or modified CDS," "mRNA product design a-1 comprising a derived and / or modified sequence," or "mRNA product design b-4 comprising a derived and / or modified sequence," or "mRNA product design c-5 comprising a derived and / or modified sequence," or "mRNA product design g-4 comprising a derived and / or modified sequence," etc.) and whether the sequence comprises or consists of a wild-type sequence ("wt") or whether the sequence comprises or consists of a sequence-optimized sequence (e.g., "opt1," "opt2," "opt3," "opt4," "opt5," "opt6," "opt11"; sequence optimization is described in more detail below). For example, the numeric identifier of SEQ ID NO: 68 <223> is given under<SEQUENCE_DESCRIPTOR> is: "derived and / or modified protein sequence (artificial)". Numeric identifier <223> is given under<CONSTRUCT_IDENTIFIER> has the following structure: ("organism_construct name" or "organism_accession number_construct name") and is intended to help those skilled in the art unambiguously derive suitable nucleic acid sequences (e.g., RNA, mRNA) encoding the same RSV protein of the present invention. For example, the numerical identifier of SEQ ID NO: 68 <223> is given under<CONSTRUCT_IDENTIFIER> is "HRSV(A2)_F0". In this example, each protein sequence is derived from "HRSV(A2)" (organism) and the protein contains the structural element "F0" (construct name, full-length F).When a person skilled in the art uses the construct identifier of SEQ ID NO: 68, i.e., "HRSV(A2)_F0", said person easily arrives at a list of suitable nucleic acid coding sequences, such as RNA coding sequences and mRNA sequences, which can be easily obtained from the sequence listing of the present invention.

[0307] Composition: A second aspect relates to a composition comprising at least one artificial RNA of the first aspect.

[0308] In a preferred embodiment of the second aspect, the composition comprises at least one artificial RNA of the first aspect and, optionally, at least one pharmaceutically acceptable carrier.

[0309] As used herein, the term "pharmaceutically acceptable carrier" preferably includes a liquid or non-liquid base for the composition. When the composition is provided in liquid form, the carrier is preferably water, typically pyrogen-free water; isotonic saline or a buffered (aqueous) solution, such as a phosphate or citrate buffer solution. Water or a buffer, preferably an aqueous buffer, can be used, which contains sodium salts, preferably at least 50 mM sodium salts, calcium salts, preferably at least 0.01 mM calcium salts, and optionally potassium salts, preferably at least 3 mM potassium salts. According to a preferred embodiment, the sodium salts, calcium salts, and optionally potassium salts may be present in the form of their halides (e.g., chlorides, iodides, or bromides), hydroxides, carbonates, bicarbonates, or sulfates. Examples of sodium salts include, but are not limited to, NaCl, NaI, NaBr, Na2CO3, NaHCO3, and Na2SO4, examples of potassium salts include, but are not limited to, KCl, KI, KBr, K2CO3, KHCO3, and K2SO4, and examples of calcium salts include, but are not limited to, CaCl2, CaI2, CaBr2, CaCO3, CaSO4, and Ca(OH)2. Additionally, organic anions of the above cations may be contained in the buffer solution.

[0310] In some embodiments, the compositions defined herein may comprise a plurality of or at least more than one artificial RNA as defined in the context of the first or second aspect of the invention.

[0311] In some embodiments, at least one RNA included in the composition is a bi- or multicistronic nucleic acid, particularly a bi- or multicistronic nucleic acid as defined herein, that encodes at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 different antigenic peptides or proteins from the same and / or different RSVs.

[0312] In an embodiment, the composition can include at least two, three, four, five, six, seven, eight, nine, ten, or more different artificial RNAs, as defined in the context of the first aspect of the present invention, each encoding at least one antigenic peptide or protein derived from a genetically identical RSV or a fragment or variant thereof. The term "identical" or "identical RSV" used in the context of a virus, e.g., "identical virus," should be understood to mean genetically identical. In particular, the (genetically) identical virus expresses identical proteins or peptides, and all proteins or peptides have the same amino acid sequence. In particular, the (genetically) identical RSV expresses essentially identical proteins, peptides, or polyproteins, and these proteins, peptides, or polyproteins preferably do not differ in their amino acid sequence. A non-limiting list of exemplary RSV viruses is shown in List 1.

[0313] In some embodiments, the composition can include at least two, three, four, five, six, seven, eight, nine, ten, or more different RNAs, as defined in the context of the first aspect of the invention, each encoding at least one peptide or protein derived from a genetically different RSV or a fragment or variant thereof. The term "different" or "different RSVs," as used in the context of a virus, e.g., "different viruses," should be understood as a difference between at least two respective viruses, where this difference is expressed in the RNA genome of each respective virus. In particular, the (genetically) different RSVs express at least one different protein, peptide, or polyprotein, and the at least one different protein, peptide, or polyprotein preferably differs by at least one amino acid.

[0314] In other embodiments, the composition comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more additional RNA constructs encoding a RSV antigen selected from glycoprotein G, short hydrophobic protein SH, matrix protein M, nucleoprotein N, large polymerase L, M2-1 protein, M2-2 protein, phosphoprotein P, nonstructural protein NS1, or nonstructural protein NS2, or any combination thereof.

[0315] For the preparation of compositions comprising at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 additional RNA constructs encoding RSV, it is preferred to use, and adapt as appropriate, the methods disclosed in PCT application PCT / EP2016 / 082487 or published patent application WO 2017 / 1090134A1.

[0316] In a preferred embodiment, the composition of the second embodiment comprises at least one artificial RNA of the first embodiment and at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein from RSV selected from matrix protein M, nucleoprotein N, M2-1 protein, and / or phosphoprotein P, or any combination thereof. The addition of the additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein from RSV M, N, M2-1, and / or phosphoprotein P (or a combination thereof) can, for example, enhance a T cell immune response.

[0317] It should be noted that embodiments relating to the artificial RNA of the first aspect also apply to at least one further artificial RNA of the second aspect and may be understood as preferred embodiments (e.g., embodiments relating to UTR combinations, CDS optimization, histone stem loops, polyA, polyC, cap structures, mRNA structure, mRNA production, mRNA purification, etc.).

[0318] Thus, the composition of the second aspect may suitably comprise at least one artificial RNA of the first aspect encoding RSF F, and at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from the RSV matrix protein M.

[0319] Alternatively, the composition of the second aspect may suitably comprise at least one artificial RNA of the first aspect encoding RSF F and at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV N.

[0320] Alternatively, the composition of the second aspect may suitably comprise at least one artificial RNA of the first aspect encoding RSF F and at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M2-1.

[0321] Alternatively, the composition of the second aspect may suitably comprise at least one artificial RNA of the first aspect encoding RSF F and at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV P.

[0322] In a preferred embodiment, the composition of the second aspect comprises two additional artificial RNA species, each comprising at least one coding sequence encoding at least one antigenic peptide or protein from RSV selected from matrix protein M, nucleoprotein N, M2-1, and phosphoprotein P.

[0323] Thus, the composition of the second aspect may suitably comprise at least one artificial RNA of the first aspect encoding RSF F, at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M, and at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV N.

[0324] Thus, the composition of the second aspect may suitably comprise at least one artificial RNA of the first aspect encoding RSF F, at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV N, and at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M2-1.

[0325] Thus, the composition of the second aspect may suitably comprise at least one artificial RNA of the first aspect encoding RSF F, at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV P, and at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M2-1.

[0326] Thus, the composition of the second aspect may suitably comprise at least one artificial RNA of the first aspect encoding RSF F, at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV P, and at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV N.

[0327] Thus, the composition of the second aspect may suitably comprise at least one artificial RNA of the first aspect encoding RSF F, at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M, and at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M2-1.

[0328] Thus, the composition of the second aspect may suitably comprise at least one artificial RNA of the first aspect encoding RSF F, at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M, and at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV P.

[0329] In a preferred embodiment, the composition of the second aspect comprises three additional artificial RNA species, each comprising at least one coding sequence encoding at least one antigenic peptide or protein from RSV selected from matrix protein M, nucleoprotein N, M2-1, and phosphoprotein P.

[0330] Thus, the composition of the second aspect may suitably comprise at least one artificial RNA of the first aspect encoding RSF F, at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M, at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV N, and at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV P.

[0331] Thus, the composition of the second aspect may comprise at least one artificial RNA of the first aspect encoding RSF F, at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M2-1, at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV N, and at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV P.

[0332] Thus, the composition of the second aspect may comprise at least one artificial RNA of the first aspect encoding RSF F, at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M2-1, at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M, and at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV P.

[0333] Thus, the composition of the second aspect may comprise at least one artificial RNA of the first aspect encoding RSF F, at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M2-1, at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M, and at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV N.

[0334] Thus, the composition of the second aspect may suitably comprise at least one artificial RNA of the first aspect encoding RSF F and at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M2-1.

[0335] In a preferred embodiment, the composition of the second embodiment comprises four additional artificial RNA species, each comprising at least one coding sequence encoding at least one antigenic peptide or protein from RSV selected from matrix protein M, nucleoprotein N, M2-1, and phosphoprotein P.

[0336] Thus, the composition of the second aspect may comprise at least one artificial RNA of the first aspect encoding RSF F, at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M, at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV N, at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV P, and at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV M2-1.

[0337] In a preferred embodiment of the second aspect, the coding sequence of the additional artificial RNA encodes at least one amino acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any of SEQ ID NOs: 9684, 10053-10133, 10134, 10503-10636, 10637, 11006-11182, 11183, 11552-11725, 19844, 20213, 20582, 20951, or a fragment or variant of any of these sequences. Further information regarding each of these preferred amino acid sequences encoding RSV proteins can be found in the Sequence Listing, in particular the identifiers 10053-10133, 10134, 10503-10636, 10637, 11006-11182, 11183, 11552-11725, 19844, 20213, 20582, 20951, or a fragment or variant of any of these sequences. <223> This can also be seen in the details provided below.

[0338] In a further embodiment of the second aspect, the coding sequence of the further artificial RNA encodes at least one antigenic peptide or protein as defined herein and further a heterologous secretory signal sequence or a heterologous secretory signal peptide, which is capable of increasing secretion of the encoded antigenic peptide or protein.

[0339] Suitable secretory signal peptides can be selected from the list of amino acid sequences according to SEQ ID NOs: 1 to 1115 and 1728 of WO 2017 / 081082, or fragments or variants of these sequences. At the nucleic acid level, any nucleic acid sequence (e.g., RNA sequence) encoding such an amino acid sequence can be selected. In this context, the disclosure of WO 2017 / 081082 is incorporated herein by reference.

[0340] According to some embodiments, the secretory signal sequence comprises an amino acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any of SEQ ID NOs: 21329-21362, or a fragment or variant of any of these sequences. Further information regarding each of these suitable amino acid sequences encoding secretory signal sequences can be found in the Sequence Listing, in particular, those identified by the identifiers <223> This can also be seen in the details provided below.

[0341] In a preferred embodiment, the additional artificial RNA of the composition is (a) at least one heterologous 5' untranslated region (5'-UTR) and / or at least one heterologous 3' untranslated region (3'-UTR); and (b) encoding at least one antigenic peptide or protein derived from RSV M, N, M2-1, and / or phosphoprotein P, or a fragment or variant thereof, operably linked to the 3'-UTR and / or 5'-UTR, preferably comprising an amino acid sequence selected from any of SEQ ID NOs: 9684, 10053-10133, 10134, 10503-10636, 10637, 11006-11182, 11183, 11552-11725, 19844, 20213, 20582, and 20951, or a fragment or variant of any of these sequences.

[0342] In a preferred embodiment, the additional artificial RNA of the composition is (a) at least one heterologous 5' untranslated region (5'-UTR) and / or at least one heterologous 3' untranslated region (3'-UTR); and (b) encoding at least one antigenic peptide or protein derived from RSV M2, or a fragment or variant thereof, operably linked to the 3'-UTR and / or 5'-UTR, preferably comprising an amino acid sequence selected from SEQ ID NOs: 11183, 20953, and 21414, or at least one coding sequence encoding a fragment or variant of any of these sequences.

[0343] Preferably, the further artificial RNA of the composition comprises a coding sequence operably linked to a 3'-UTR and a 5'-UTR selected from a-1, a-2, a-3, a-4, a-5, b-1, b-2, b-3, b-4, b-5, c-1, c-2, c-3, c-4, c-5, d-1, d-2, d-3, d-4, d-5, e-1, e-2, e-3, e-4, e-5, e-6, f-1, f-2, f-3, f-4, f-5, g-1, g-2, g-3, g-4, g-5, h-1, h-2, h-3, h-4, h-5, i-1, i-2 or i-3 (as defined in the context of the first aspect).

[0344] Preferably, the further artificial RNA of the composition has at least one or more uracil nucleotides, or preferably all uracil nucleotides, substituted with pseudouridine (Ψ) or N1-methylpseudouridine (m1Ψ) nucleotides.

[0345] Thus, in other embodiments, the further artificial RNA of the composition may comprise a 5' cap sequence element according to SEQ ID NO: 43 or 21321, or a fragment or variant thereof.

[0346] In a preferred embodiment of the second aspect, the coding sequence of the further RNA comprises at least one nucleic acid sequence identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NOs: 9685-9692, 10135-10142, 10638-10645, 11184-11119, 19845-19852, 20214-20221, 20583-20590, 20952-20959, 21385-21388, 21411-21414 or a fragment or variant (cds optimized as defined in the context of the first aspect) of any of these sequences. Further information regarding each of these preferred amino acid sequences encoding RSV proteins can be found in the Sequence Listing, particularly the identifiers AA, ... <223> This can also be seen in the details provided below.

[0347] In a preferred embodiment of the second aspect, the further artificial RNA, preferably the further mRNA, of the composition comprises, preferably in the 5' to 3' direction, the following elements a) to i): a) a 5'-cap structure, preferably a 5'-cap structure as described in the context of the first embodiment, most preferably a Cap1 structure; b) optionally, a 5'-UTR as described in the context of the first aspect, preferably at least one selected from SEQ ID NOs: 1 to 22; c) a ribosome binding site, preferably a ribosome binding site described herein; d) at least one coding sequence selected from SEQ ID NOs: 9685-9692, 10135-10142, 10638-10645, 11184-11119, 19845-19852, 20214-20221, 20583-20590, 20952-20959, 21385-21388, 21411-21414; e) a 3'-UTR as described in the context of the first aspect, preferably at least one selected from SEQ ID NOs: 23 to 38; f) optionally a poly(A) sequence, preferably a poly(A) sequence as described in the context of the first aspect; g) optionally a poly(C) sequence, preferably a poly(C) sequence as described in the context of the first aspect; h) optionally a histone stem loop, preferably a histone stem loop as described in the context of the first aspect; i) optionally a 3'-end sequence element as described in the context of the first aspect, preferably a 3'-end sequence element according to SEQ ID NOs: 44 to 63, 21322 to 21328; and optionally, at least one or more, preferably all, uracil nucleotides are substituted with pseudouridine (Ψ) or N1-methylpseudouridine (m1Ψ) nucleotides.

[0348] In a particularly preferred embodiment of the second aspect, the further artificial RNA, preferably the further mRNA, of the composition comprises, preferably in the 5' to 3' direction, the following elements a) to i): a) a 5'-cap structure, preferably a 5'-cap structure as described in the context of the first embodiment, most preferably a Cap1 structure; b) optionally, a 5'-UTR as described in the context of the first aspect, preferably at least one selected from SEQ ID NOs: 1 to 22; c) a ribosome binding site, preferably a ribosome binding site described herein; d) at least one coding sequence selected from SEQ ID NOs: 11185-11191, 21388, 20952-20959, 21414; e) a 3'-UTR as described in the context of the first aspect, preferably at least one selected from SEQ ID NOs: 23 to 38; f) optionally a poly(A) sequence, preferably a poly(A) sequence as described in the context of the first aspect; g) optionally a poly(C) sequence, preferably a poly(C) sequence as described in the context of the first aspect; h) optionally a histone stem loop, preferably a histone stem loop as described in the context of the first aspect; i) optionally a 3'-end sequence element as described in the context of the first aspect, preferably a 3'-end sequence element according to SEQ ID NOs: 44 to 63, 21322 to 21328; and optionally, at least one or more, preferably all, uracil nucleotides are substituted with pseudouridine (Ψ) or N1-methylpseudouridine (m1Ψ) nucleotides.

[0349] In that context, preferred RSV polypeptide, nucleic acid, and mRNA sequences of the second embodiment are shown in Table 7A.

[0350] In Table 7A, columns A through D show specific preferred constructs of the second embodiment derived from RSV proteins: column A shows preferred sequences for M, column B shows preferred sequences for N, column C shows preferred sequences for P, and column D shows preferred sequences for M2-1.

[0351] Protein names / designs are listed in row 1 (columns A-D), and protein SEQ ID NOs as listed in the sequence listing are listed in row 2 ("PRT"). Preferred protein SEQ ID NOs for HRSV(A2) are listed in row 3 ("HRSV(A2)PRT"). SEQ ID NOs for the corresponding coding sequences of each listed HRSV(A2) protein are listed in rows 4-11 ("cds wt," "cds opt1," "cds opt2," "cds opt3," "cds opt4," "cds opt5," "cds opt6," and "cds opt11," respectively). SEQ ID NOs for corresponding mRNA constructs comprising the coding sequences and preferred 3'-UTRs and 5'-UTRs of the present invention are listed in rows 12-55 ("mRNA Design a-1" to "mRNA Design i-3" as described herein).

[0352] Preferred protein SEQ ID NOs for HRSV (Memphis-37) are listed in column 56 ("Memphis-37 PRT"). SEQ ID NOs for the corresponding coding sequences for each of the listed HRSV (Memphis-37) proteins are listed in rows 57-64 ("cds wt," "cds opt1," "cds opt2," "cds opt3," "cds opt4," "cds opt5," "cds opt6," and "cds opt11," respectively). SEQ ID NOs for corresponding mRNA constructs comprising the coding sequences and suitable 3'-UTRs and 5'-UTRs of the present invention are listed in rows 65-108 ("mRNA Design a-1" through "mRNA Design i-3" as described herein).

[0353] Further information regarding the type of coding sequence contained in the indicated mRNA constructs can be found in the sequence listing for each SEQ ID NO: <223> Shown in the identifier.

[0354] [Table 7A-1] [Table 7A-2] [Table 7A-3]

[0355] Thus, in a preferred embodiment, the composition of the second aspect comprises at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein from RSV selected from matrix protein M, nucleoprotein N, M2-1 protein, phosphoprotein P, wherein the additional artificial RNA is selected from SEQ ID NOs: 9693-10052, 10143-10502, 10646-11005, 11192-11551, 19853-20212, 20222-20581, 20591-20950, 20960-21319, 21481-21482 488, 21627-21634, 21553-21560, 21699-21706, or a fragment or variant of any of these sequences, optionally comprising or consisting of an RNA sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid sequence selected from the group consisting of: 488, 21627-21634, 21553-21560, 21699-21706, or a fragment or variant of any of these sequences, optionally with at least one, more than one, or all uracil nucleotides substituted with pseudouridine (Ψ) nucleotides or N1-methylpseudouridine (m1Ψ) nucleotides. Further information regarding each of these preferred amino acid sequences encoding RSV proteins can be found in the Sequence Listing, particularly those identified by the identifiers 488, 21627-21634, 21553-21560, and 21699-21706, as described below. <223> This can also be seen in the details provided below.

[0356] In a particularly preferred embodiment of the second aspect, the composition of the second aspect comprises: - Sequence numbers 78 to 482, 493 to 897, 907 to 1266, 1276 to 1635, 1645 to 2004, 2014 to 2373, 2383 to 2742, 2752 to 3111, 3121 to 3480, 3490 to 3849, 3859 to 4218, 4228 to 4587, 4597 to 4956, 4966 to 5325 , 5335~5694, 5704~6063, 6073~6432, 6442~6801, 6811~7170, 7180~7539, 7549~7908, 7918~8277, 8278, 11735~12094, 12104~12463, 12473~12832, 12842~13201, 1394 9~14308, 14318~14677, 14687~15046, 15056~15415, 15425~15784, 15794~16153, 13211~13570, 13580~13939, 16163~16522, 16532~16891, 16901~17260, 17270~1762 9, 17639 to 17998, 18008 to 18367, 18377 to 18736, 18746 to 19105, 19115 to 19474, 19484 to 19843, 21415 to 21480, 21561 to 21626, 21489 to 21552, 21635 to 21698 (RSV as defined in the first embodiment) At least one artificial RNA that is identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 9693-10052, 19853-20212, 21481, 21482, 21627, 21628, 21553, 21554, 21699, and 21700 (encoding RSV F). an additional artificial RNA that is identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid sequence selected from the group consisting of: a nucleic acid sequence encoding M; - Sequence numbers 78 to 482, 493 to 897, 907 to 1266, 1276 to 1635, 1645 to 2004, 2014 to 2373, 2383 to 2742, 2752 to 3111, 3121 to 3480, 3490 to 3849, 3859 to 4218, 4228 to 4587, 4597 to 4956, 4966 to 5325 , 5335~5694, 5704~6063, 6073~6432, 6442~6801, 6811~7170, 7180~7539, 7549~7908, 7918~8277, 8278, 11735~12094, 12104~12463, 12473~12832, 12842~13201, 1394 9~14308, 14318~14677, 14687~15046, 15056~15415, 15425~15784, 15794~16153, 13211~13570, 13580~13939, 16163~16522, 16532~16891, 16901~17260, 17270~1762 9, 17639 to 17998, 18008 to 18367, 18377 to 18736, 18746 to 19105, 19115 to 19474, 19484 to 19843, 21415 to 21480, 21561 to 21626, 21489 to 21552, 21635 to 21698 (RSV as defined in the first embodiment) At least one artificial RNA that is identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 10143 to 10502, 20222 to 20581, 21483, 21484, 21629, 21630, 21555, 21556, 21701, and 21702 (RSV encoding RSV F). an additional artificial RNA that is identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid sequence selected from the group consisting of: a nucleic acid sequence encoding N; or - Sequence numbers 78 to 482, 493 to 897, 907 to 1266, 1276 to 1635, 1645 to 2004, 2014 to 2373, 2383 to 2742, 2752 to 3111, 3121 to 3480, 3490 to 3849, 3859 to 4218, 4228 to 4587, 4597 to 4956, 4966 to 5325 , 5335~5694, 5704~6063, 6073~6432, 6442~6801, 6811~7170, 7180~7539, 7549~7908, 7918~8277, 8278, 11735~12094, 12104~12463, 12473~12832, 12842~13201, 1394 9~14308, 14318~14677, 14687~15046, 15056~15415, 15425~15784, 15794~16153, 13211~13570, 13580~13939, 16163~16522, 16532~16891, 16901~17260, 17270~1762 9, 17639 to 17998, 18008 to 18367, 18377 to 18736, 18746 to 19105, 19115 to 19474, 19484 to 19843, 21415 to 21480, 21561 to 21626, 21489 to 21552, 21635 to 21698 (RSV as defined in the first embodiment) At least one artificial RNA that is identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 10646-11005, 20591-20950, 21485, 21486, 21631, 21632, 21557, 21558, 21703, and 21704 (encoding RSV F). at least one additional artificial RNA that is identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid sequence selected from the group consisting of: a nucleic acid sequence encoding P; or - Sequence numbers 78 to 482, 493 to 897, 907 to 1266, 1276 to 1635, 1645 to 2004, 2014 to 2373, 2383 to 2742, 2752 to 3111, 3121 to 3480, 3490 to 3849, 3859 to 4218, 4228 to 4587, 4597 to 4956, 4966 to 5325 , 5335~5694, 5704~6063, 6073~6432, 6442~6801, 6811~7170, 7180~7539, 7549~7908, 7918~8277, 8278, 11735~12094, 12104~12463, 12473~12832, 12842~13201, 1394 9~14308, 14318~14677, 14687~15046, 15056~15415, 15425~15784, 15794~16153, 13211~13570, 13580~13939, 16163~16522, 16532~16891, 16901~17260, 17270~1762 9, 17639 to 17998, 18008 to 18367, 18377 to 18736, 18746 to 19105, 19115 to 19474, 19484 to 19843, 21415 to 21480, 21561 to 21626, 21489 to 21552, 21635 to 21698 (RSV as defined in the first embodiment) At least one artificial RNA that is identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 11192 to 11551, 20960 to 21319, 21487, 21488, 21633, 21634, 21559, 21560, 21705, and 21706 (encoding RSV F). at least one additional artificial RNA that is identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid sequence selected from the group consisting of: a nucleic acid sequence encoding M2-1; or - Sequence numbers 78 to 482, 493 to 897, 907 to 1266, 1276 to 1635, 1645 to 2004, 2014 to 2373, 2383 to 2742, 2752 to 3111, 3121 to 3480, 3490 to 3849, 3859 to 4218, 4228 to 4587, 4597 to 4956, 4966 to 5325 , 5335~5694, 5704~6063, 6073~6432, 6442~6801, 6811~7170, 7180~7539, 7549~7908, 7918~8277, 8278, 11735~12094, 12104~12463, 12473~12832, 12842~13201, 1394 9~14308, 14318~14677, 14687~15046, 15056~15415, 15425~15784, 15794~16153, 13211~13570, 13580~13939, 16163~16522, 16532~16891, 16901~17260, 17270~1762 9, 17639 to 17998, 18008 to 18367, 18377 to 18736, 18746 to 19105, 19115 to 19474, 19484 to 19843, 21415 to 21480, 21561 to 21626, 21489 to 21552, 21635 to 21698 (RSV as defined in the first embodiment) At least one artificial RNA that is identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 9693-10052, 19853-20212, 21481, 21482, 21627, 21628, 21553, 21554, 21699, and 21700 (encoding RSV F). At least one additional artificial RNA identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 10646-11005, 20591-20950, 21485, 21486, 21631, 21632, 21557, 21558, 21703, 21704 (encoding RSV M)an additional artificial RNA that is identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid sequence selected from the group consisting of: a nucleic acid sequence encoding P; - Sequence numbers 78 to 482, 493 to 897, 907 to 1266, 1276 to 1635, 1645 to 2004, 2014 to 2373, 2383 to 2742, 2752 to 3111, 3121 to 3480, 3490 to 3849, 3859 to 4218, 4228 to 4587, 4597 to 4956, 4966 to 5325 , 5335~5694, 5704~6063, 6073~6432, 6442~6801, 6811~7170, 7180~7539, 7549~7908, 7918~8277, 8278, 11735~12094, 12104~12463, 12473~12832, 12842~13201, 1394 9~14308, 14318~14677, 14687~15046, 15056~15415, 15425~15784, 15794~16153, 13211~13570, 13580~13939, 16163~16522, 16532~16891, 16901~17260, 17270~1762 9, 17639 to 17998, 18008 to 18367, 18377 to 18736, 18746 to 19105, 19115 to 19474, 19484 to 19843, 21415 to 21480, 21561 to 21626, 21489 to 21552, 21635 to 21698 (RSV as defined in the first embodiment) At least one artificial RNA that is identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 9693-10052, 19853-20212, 21481, 21482, 21627, 21628, 21553, 21554, 21699, and 21700 (encoding RSV F). At least one additional artificial RNA identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 10646-11005, 20591-20950, 21485, 21486, 21631, 21632, 21557, 21558, 21703, 21704 (encoding RSV M)Further artificial RNAs identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the nucleic acid sequence selected from the group consisting of SEQ ID NOs: 10143-10502, 20222-20581, 21483, 21484, 21629, 21630, 21555, 21556, 21701, 21702 (encoding RSV P) an additional artificial RNA that is identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid sequence selected from the group consisting of: a nucleic acid sequence encoding N; or - Sequence numbers 78 to 86, 21415 to 21417, 21561 to 21563, 474 to 482, 11735 to 11742, 21489, 21490, 21635, 21636, 12087 to 12094, 493 to 501, 21418 to 21420, 21564 to 21566, 88 9~897, 12104~12111, 21491, 21492, 21637, 21638, 12456~12463, 907~914, 21421~21423, 21567~21569, 1259~1266, 12473~12480, 21493~21495, 2163 9~21641, 12825~12832, 1276~1283, 21424~21426, 21570~21572, 1628~1635, 12842~12849, 21496~21498, 21642~21644, 13194~13201, 1645~1652, 21 433~21435, 21579~21581, 1997~2004, 13949~13956, 21505~21507, 21651~21653, 14301~14308, 2014~2021, 21436~21438, 21582~21584, 2366~2373, 14318~14325, 21508~21510, 21654~21656, 14670~14677, 2383~2390, 21439~21441, 21585~21587, 2735~2742, 14687~14694, 21511~21513, 21657~21 659, 15039~15046, 2752~2759, 21442~21444, 21588~21590, 3104~3111, 15056~15063, 21514~21516, 21660~21662, 15408~15415, 3121~3128, 21445~ 21447, 21591~21593, 3473~3480, 15425~15432, 21517~21519, 21663~21665, 15777~15784, 3490~3497, 21448~21450, 21594~21596, 3842~3849, 1579 4~15801, 21520~21522, 21666~21668, 16146~16153, 3859~3866, 21427~21429, 21573~21575, 4211~4218, 13211~13218, 21499~21501, 21645~21647,13563~13570, 4228~4235, 21430~21432, 21576~21578, 4580~4587, 13580~13587, 21502~21504, 21648~21650, 13932~13939, 4597~4604, 21451~21453, 21597~21599, 4949~4956, 16163~16170, 21523~21525, 21669~21671, 16515~16522, 4966~4973, 21454~21456, 21600~21602, 5318~5325, 1653 2 to 16539, 21526 to 21528, 21672 to 21674, 16884 to 16891, 5335 to 5342, 21457 to 21459, 21603 to 21605, 5687 to 5694, 16901 to 16908, 21529 to 21531, 21675 to 21677, 17253 to 17260, 5704 to 5711, 21460 to 21462, 21606 to 21608, 6056 to 6063, 17270 to 17277, 21532 to 21534, 21678 to 21680, 17622 to 17629 (RSV as defined in the first embodiment) At least one artificial RNA that is identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 11192 to 11551, 20960 to 21319, 21487, 21488, 21633, 21634, 21559, 21560, 21705, and 21706 (encoding RSV F). at least one additional artificial RNA that is identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid sequence selected from the group consisting of: a nucleic acid sequence encoding M2-1; and optionally, at least one, or more than one, or all uracil nucleotides are substituted with pseudouridine (Ψ) nucleotides or N1-methylpseudouridine (m1Ψ) nucleotides.

[0357] In various embodiments, the RSV F (F0, F-del, F0_DSCav1, F-del_DSCav1, F_DSCav1_mut0, F-del_DSCav1_mut0, F_DSCav1_mut1, F-del_DSCav1_mut1, F_DSCav1_mut2, F-del_DSCav1_mut2, F_DSCav1_mut3, F-del_DSCav1_mut3, F_DSCav1_mut4, F-del_DSCav1_mut4, F_DSCav1_mut5, F-del_DSCav1_mut5, F_DSCav1_mut6, F-del_DSCav1_mut6, F_DSCav1_mut7, F-del_DSCav1_mut7, F_DSCav1_mut8, F-del_DSCav1_mut8) RNA construct and RSV Various combinations of T-cell antigen RNA constructs (RSV M, N, M2-1, or P) are suitable for inclusion in the compositions (disclosed in Table 7B; Combinations 1-64). Combinations 49-64 are preferred, with Combination 64 being particularly preferred.

[0358] [Table 7B-1] [Table 7B-2]

[0359] Table 7B discloses compositions of a second aspect, comprising at least one RNA encoding RSV F from strain A2 as defined in the first aspect, and at least one additional artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein from RSV from strain A2 selected from matrix protein M, nucleoprotein N, M2-1 protein, and phosphoprotein P. In a further embodiment of the invention, any of the disclosed compositions in Table 7B is also applicable to antigenic peptides or proteins from RSV isolate Memphis-37 (strain Memphis-37).

[0360] In a particularly preferred embodiment, the composition comprises one RSV F RNA construct (F0, F-del, F0_DSCav1, F-del_DSCav1, F_DSCav1_mut0, F-del_DSCav1_mut0, F_DSCav1_mut1, F-del_DSCav1_mut1, F_DSCav1_mut2, F-del_DSCav1_mut2, F_DSCav1_mut3, F-del_DSCav1_mut3, F_DSCav1_mut4, F-del_DSCav1_mut4, F_DSCav1_mut5, F-del_DSCav1_mut5, F_DSCav1_mut6, F-del_DSCav1_mut6, F_DSCav1_mut7, F-del_DSCav1_mut7, F_DSCav1_mut8, F-del_DSCav1_mut8) and one RSV M2-1 RNA construct, preferably comprising F-del_DSCav1_mut5 and M2-1.

[0361] In a particularly preferred embodiment, an RNA construct encoding an RSV F selected from SEQ ID NOs: 5704-5711, 21460-21462, 21606-21608, 17270-17277, 21532-21534, 21678-21680 and an RNA construct encoding M2-1 selected from SEQ ID NOs: 11192-11199, 21487, 21488, 21633, 21634, 20960-20967, 21559, 21560, 21705, 21706 are included in the composition of the invention.

[0362] In various embodiments, at least one artificial RNA of the first aspect and at least one additional artificial RNA described herein are derived from the same RSV (e.g., any virus selected from List 1).

[0363] In a preferred embodiment, at least one artificial RNA of the first aspect is derived from HRSV(A2), and at least one further artificial RNA described herein is derived from HRSV(A2).

[0364] In a preferred embodiment, at least one artificial RNA of the first aspect is derived from HRSV(Memphis-37), and at least one further artificial RNA described herein is derived from HRSV(Memphis-37).

[0365] In the context of the present invention, it should be understood that specific combinations of coding sequences can be generated by any combination of monocistronic, bicistronic, and multicistronic artificial nucleic acids and / or multi-antigen constructs / nucleic acids to obtain nucleic acid compositions encoding multiple antigenic peptides or proteins as defined herein.

[0366] In addition, one or more compatible solid or liquid fillers, diluents, or encapsulating compounds suitable for human administration can also be used. As used herein, the term "compatible" means that the components of the composition can be mixed with at least one RNA of the composition and optionally with additional artificial RNAs under normal use conditions so that no interactions occur that could substantially reduce the biological activity or efficacy of the composition. Pharmaceutically acceptable carriers, fillers, and diluents must be of sufficiently high purity and sufficiently low toxicity to make them suitable for administration to a subject to be treated. Compounds that can be used as pharmaceutically acceptable carriers, fillers, or components thereof include, for example, sugars such as lactose, glucose, trehalose, and sucrose; starches such as corn starch or potato starch; dextrose; cellulose and its derivatives, such as sodium carboxymethylcellulose, ethyl cellulose, cellulose acetate, and the like; powdered tragacanth; malt; gelatin; tallow; solid glidants, such as stearic acid, magnesium stearate; calcium sulfate; vegetable oils, such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil, and cocoa oil; polyols, such as polypropylene glycol, glycerol, sorbitol, mannitol, and polyethylene glycol; alginic acid, and the like.

[0367] Further additives that may be included in the composition are emulsifiers, such as Tween; wetting agents, such as sodium lauryl sulfate; coloring agents; flavoring agents, pharmaceutical carriers; tableting agents; stabilizers; antioxidants; and preservatives.

[0368] complex In a preferred embodiment of the second aspect, at least one artificial RNA of the first aspect and, optionally, further artificial RNAs of the second aspect are complexed or associated or at least partially complexed or associated with one or more cationic or polycationic compounds, preferably cationic or polycationic polymers, cationic or polycationic polysaccharides, cationic or polycationic lipids, cationic or polycationic proteins, cationic or polycationic peptides, or any combination thereof.

[0369] The term "cationic or polycationic compound" as used herein is recognized and understood by those skilled in the art and is intended to mean a positively charged molecule, e.g., at a pH of about 1-9, about 3-8, about 4-8, about 5-8, more preferably about 6-8, more preferably about 7-8, and most preferably at physiological pH, e.g., about 7.2 to about 7.5. Thus, a cationic component, e.g., a cationic peptide, cationic protein, cationic polymer, cationic polysaccharide, or cationic lipid, can be any positively charged compound or polymer that is positively charged under physiological conditions. A "cationic or polycationic peptide or protein" can contain at least one positively charged amino acid, or more than one positively charged amino acid, e.g., an amino acid selected from Arg, His, Lys, or Orn. ​​Thus, a "polycationic" component also falls within the scope of exhibiting a positive charge of more than one under given conditions.

[0370] Particularly preferred cationic or polycationic compounds in this context can be selected from the following list of cationic or polycationic peptides or proteins fragments thereof: protamine, nucleolin, spermine, or spermidine, or other cationic peptides or proteins, such as poly-L-lysine (PLL), polyarginine, basic polypeptides, cell penetrating peptides (CPPs), such as HIV-binding peptides, HIV-1 Tat (HIV), Tat-derived peptides, penetratin, VP22-derived or analogue peptides, HSV More preferably, the nucleic acid as defined herein, preferably the mRNA as defined herein, is complexed with one or more polycations, preferably protamine or oligofectamine, most preferably protamine.

[0371] In a preferred embodiment of the second aspect, at least one artificial RNA of the first aspect and optionally further artificial RNAs of the second aspect are complexed with protamine.

[0372] Further preferred cationic or polycationic compounds which can be used as transfection or complexing agents are cationic polysaccharides, such as chitosan, polybrene, etc.; cationic lipids, such as DOTMA, DMRIE, di-C14-amidine, DOTIM, SAINT, DC-Chol, BGTC, CTAP, DOPC, DODAP, DOPE: dioleylphosphatidylethanolamine, DOSPA, DODAB, DOIC, DMEPC, DOGS, DIMRI, DOTAP, DC-6-14, CLIP1, CLIP6, CLIP9, oligofectamine, etc.; or cationic or polycationic polymers, such as modified polyamino acids, such as beta-amino acid polymers or inverse polyamides, modified polyethylenes, such as PVP, modified acrylates, such as pDMAEMA, pAM, etc. modified amidoamines such as AM; modified polybeta-aminoesters (PBAEs) such as diamine-terminated 1,4 butanediol diacrylate-co-5-amino-1-pentanol polymers; dendrimers such as polypropyleneamine dendrimers or pAMAM-based dendrimers; polyimines such as PEI and poly(propyleneimine); polyallylamine; sugar-based polymers such as cyclodextrin-based polymers and dextran-based polymers; silane-based polymers such as PMOXA-PDMS copolymers; block polymers consisting of a combination of one or more cationic blocks (e.g., selected from the cationic polymers described above) and a combination of one or more hydrophilic or hydrophobic blocks (e.g., polyethylene glycol); and the like.

[0373] In this context, it is particularly preferred that at least one artificial RNA as defined herein and optionally further artificial RNAs of the second aspect are complexed or at least partially complexed with cationic or polycationic compounds and / or polymeric carriers, preferably cationic proteins or peptides. In this context, the disclosures of WO 2010 / 037539 and WO 2012 / 113513 are incorporated herein by reference. Partially means that only a portion of the artificial nucleic acid is complexed with a cationic compound, while the remaining portion of the artificial nucleic acid (contained in the inventive (pharmaceutical) composition) is uncomplexed ("free").

[0374] In a preferred embodiment of the second aspect, at least one artificial RNA of the first aspect and, optionally, a further artificial RNA of the second aspect are complexed with one or more cationic or polycationic compounds, preferably protamine, and at least one free artificial RNA of the first aspect and, optionally, a further RNA of the second aspect.

[0375] In this context, it is particularly preferred that at least one artificial RNA as defined herein and, optionally, a further artificial RNA of the second aspect is complexed or at least partially complexed with protamine. Preferably, the molar ratio of RNA to free RNA of the nucleic acid, in particular of the protamine-complexed RNA, can be selected from a molar ratio of about 0.001:1 to about 1:0.001, including a ratio of about 1:1. Advantageously, the complexed RNA is complexed with protamine by adding a protamine-trehalose solution to the RNA sample in a 2:1 RNA:protamine weight-to-weight ratio (w / w).

[0376] Further preferred cationic or polycationic proteins or peptides that can be used for conjugation can be derived from the formula (Arg)l;(Lys)m;(His)n;(Orn)o;(Xaa)x of WO 2009 / 030481 or WO 2011 / 026641, the relevant disclosures of which are incorporated herein by reference.

[0377] In a preferred embodiment of the second aspect, at least one artificial RNA of the first aspect and optionally further artificial RNAs of the second aspect are preferably complexed or at least partially complexed with at least one cationic or polycationic protein or peptide selected from SEQ ID NOs: 64-67, 21320 or any combination thereof.

[0378] According to some embodiments, the compositions of the invention comprise RNA as defined herein and a polymeric carrier.

[0379] The term "polymeric carrier" as used herein will be recognized and understood by those skilled in the art and is intended to mean, for example, a compound that facilitates the transport and / or complexation of another compound (cargo). A polymeric carrier is typically a carrier formed from a polymer. The polymeric carrier can be associated with its cargo (nucleic acid, RNA) by covalent or non-covalent interactions.

[0380] A suitable polymeric carrier may be a polymeric carrier formed by disulfide-bridged cationic compounds. The disulfide-bridged cationic compounds may be the same or different from each other. The polymeric carrier may also contain additional components. The polymeric carrier used according to the present invention may comprise a mixture of cationic peptides, proteins, or polymers as defined herein, cross-linked by disulfide bonds (via -SH groups), and optionally additional components.

[0381] In this context, polymeric carriers according to the formula {(Arg)l;(Lys)m;(His)n;(Orn)o;(Xaa')x(Cys)y} and formula Cys,{(Arg)l;(Lys)m;(His)n;(Orn)o;(Xaa)x}Cys2 of WO 2012 / 013326 are preferred, the disclosure of which in WO 2012 / 013326 is incorporated herein by reference.

[0382] In some embodiments, the polymeric carrier that can be used to complex RNA as defined herein has the formula (LP 1 -S-[SP 2 -S] n -SP 3 -L), the disclosure of which is incorporated herein by reference.

[0383] In some embodiments, the polymeric carrier compound is formed by, comprises or consists of the peptide element CysArg12Cys (SEQ ID NO: 64) or CysArg12 (SEQ ID NO: 65) or TrpArg12Cys (SEQ ID NO: 66). In particularly preferred embodiments, the polymeric carrier compound is (R 12 C)-(R 12 C) Dimer, (WR 12 C)-(WR 12 C) dimer, or (CR 12 )-(CR 12 C)-(CR 12 ) trimers, and dimers (e.g., (WR 12 C)) or trimers (e.g., (CR 12 The individual peptide elements in are linked via -SH groups.

[0384] In a preferred embodiment of the second aspect, at least one artificial RNA of the first aspect and optionally further artificial RNAs of the second aspect are complexed or associated with a polyethylene glycol / peptide polymer comprising HO-PEG5000-S-(S-CHHHHHHRRRRHHHHHHC-S-)7-S-PEG5000-OH (SEQ ID NO: 67 as peptide monomer).

[0385] In a further preferred embodiment of the second aspect, at least one artificial RNA of the first aspect and optionally further artificial RNAs of the second aspect are complexed or associated with a polyethylene glycol / peptide polymer comprising HO-PEG5000-S-(S-CHHHHHHRRRRHHHHHHC-S-)4-S-PEG5000-OH (SEQ ID NO: 67 as peptide monomer).

[0386] In a further preferred embodiment of the second aspect, at least one artificial RNA of the first aspect and optionally further artificial RNAs of the second aspect are complexed or associated with a polyethylene glycol / peptide polymer comprising HO-PEG5000-S-(S-CGHHHHHRRRRHHHHHGC-S-)7-S-PEG5000-OH (SEQ ID NO: 21320 as peptide monomer).

[0387] In a further preferred embodiment of the second aspect, at least one artificial RNA of the first aspect and optionally further artificial RNAs of the second aspect are complexed or associated with a polyethylene glycol / peptide polymer comprising HO-PEG5000-S-(S-CGHHHHHRRRRHHHHHGC-S-)4-S-PEG5000-OH (SEQ ID NO: 21320 as peptide monomer).

[0388] In other embodiments, the composition comprises at least one artificial RNA described herein and, optionally, a further artificial RNA of the second aspect, wherein the at least one artificial RNA and, optionally, the further artificial RNA of the second aspect are complexed or associated with a polymeric carrier and, optionally, at least one lipid component de...

Claims

1. a) at least one heterologous 5' untranslated region (5'-UTR) and / or at least one heterologous 3' untranslated region (3'-UTR); b) at least one coding sequence operably linked to the 3'-UTR and / or 5'-UTR, encoding at least one antigenic peptide or protein derived from an RSV fusion (F) protein, or a fragment or variant thereof; the at least one antigenic peptide or protein derived from the RSV fusion (F) protein is a C-terminally deleted RSV F protein (F-del); the RSV fusion (F) protein comprises a DSCav1 mutation selected from S155C, S290C, S190F, and V207L; the RSV fusion (F) protein comprises two subunits, F2 and F1, in a single polypeptide chain, the F2 subunit and the F1 subunit being connected via a linker element to generate a stable F2-linker-F1 protein; the F2-linker-F1 protein does not include aa104 to aa144; Furthermore, the RSV fusion (F) protein contains at least one mutation selected from A149C and Y458C; The artificial RNA, wherein the amino acid residues and their positions of the RSV fusion (F) protein correspond to the amino acid residues and their positions of SEQ ID NO:

68.

2. The artificial RNA of claim 1, wherein the at least one heterologous 3'-UTR comprises a nucleic acid sequence derived from the 3'-UTR of a gene selected from PSMB3, ALB7, alpha-globin, CASP1, COX6B1, GNAS, NDUFA1, and RPS9, or a homolog, fragment, or variant of any of these genes.

3. The artificial RNA of any of claims 1 to 2, wherein the at least one heterologous 5'-UTR comprises a nucleic acid sequence derived from the 5'-UTR of a gene selected from HSD17B4, RPL32, ASAH1, ATP5A1, MP68, NDUFA4, NOSIP, RPL31, SLC7A3, TUBB4B, and UBQLN2, or a homolog, fragment, or variant of any of these genes.

4. The artificial RNA of any of claims 1 to 3, wherein the RSV fusion (F) protein is designed to stabilize an antigen in a pre-fusion conformation.

5. The artificial RNA described in any one of claims 1 to 4, wherein the RSV fusion (F) protein is selected from F-del_DSCav1_mut1, F-del_DSCav1_mut2, F-del_DSCav1_mut3, F-del_DSCav1_mut4, or a fragment or variant thereof.

6. The artificial RNA of any of claims 1 to 5, wherein the at least one coding sequence encodes at least one amino acid sequence identical or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to any of SEQ ID NOs: 2005, 2743, 3481, 4957, 14309, 15047, 15785, 16523, or a fragment or variant of any of these sequences.

7. The at least one coding sequence is identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NOs: 2006-2013, 2744-2751, 3482-3489, 4958-4965, 14310-14317, 15048-15055, 15786-15793, 16524-16531, 21370, 21372, 21374, 21376, 21396, 21398, 21400, 21402, or at least one amino acid sequence that is identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NOs: 2006-2013, 2744-2751, 3482-3489, 4958-4965, 14310-14317, 15048-15055, 15786-15793, 16524-16531, 21370, 21372, 21374, 21376, 21396, 21398, 21400, 21402, or a fragment or variant of any of these sequences. The artificial RNA according to any one of claims 1 to 6.

8. An artificial RNA described in any of claims 1 to 7, wherein the at least one coding sequence is a codon-modified coding sequence, and the amino acid sequence encoded by the at least one codon-modified coding sequence is unchanged compared to the amino acid sequence encoded by the corresponding wild-type coding sequence.

9. The artificial RNA described in claim 8, wherein the at least one codon-modified coding sequence is selected from a C-maximized coding sequence, a CAI-maximized coding sequence, a human codon usage adapted coding sequence, a G / C content modified coding sequence, and a G / C optimized coding sequence, or any combination thereof.

10. The artificial RNA described in any one of claims 1 to 9, wherein the artificial RNA is mRNA, self-replicating RNA, circular RNA, or replicon RNA.

11. The artificial RNA described in claim 10, wherein the artificial RNA is mRNA.

12. The artificial RNA described in any one of claims 1 to 11, wherein the artificial RNA includes a 5'-cap structure.

13. The artificial RNA described in any of claims 1 to 12, wherein the artificial RNA contains at least one poly(A) sequence and / or at least one poly(C) sequence.

14. The artificial RNA described in any of claims 1 to 13, wherein the artificial RNA comprises at least one histone stem loop, and the histone stem loop comprises a nucleic acid sequence according to SEQ ID NO: 39 or 40 or a fragment or variant thereof.

15. The artificial RNA is selected from the group consisting of SEQ ID NOs: 2014-2373, 2752-3111, 3490-3849, 4966-5325, 14318-14677, 15056-15415, 15794-16153, 16532-16891, 21436-21438, 21442-21444, 21448-21450, 21454-21456, 21508-21510, 21514-21516, 21520-21522, 21526-21528, 21582-21584, 21588-21590, 21594-21 15. The artificial RNA according to any one of claims 1 to 14, comprising or consisting of an RNA sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a nucleic acid sequence selected from the group consisting of: 596, 21600-21602, 21654-21656, 21660-21662, 21666-21668, 21672-21674, or a fragment or variant of any of these sequences.

16. The artificial RNA described in any of claims 1 to 15, wherein the at least one coding sequence encodes at least one amino acid sequence that is identical or at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:4957, or a fragment or variant of any of these sequences.

17. A composition comprising at least one artificial RNA described in any one of claims 1 to 16, optionally comprising at least one pharma- ceutically acceptable salt.

18. The composition of claim 17, comprising at least one further artificial RNA comprising at least one coding sequence encoding at least one antigenic peptide or protein derived from RSV selected from matrix protein M, nucleoprotein N, M2-1 protein, and / or phosphoprotein P, or a combination thereof.

19. A composition described in any of claims 17 to 18, wherein the at least one artificial RNA and, optionally, the further artificial RNA are complexed or associated or at least partially complexed or associated with one or more cationic or polycationic compounds.

20. The composition described in claim 19, wherein the artificial RNA and, optionally, the further artificial RNA are complexed or associated with one or more lipids to form a liposome, lipid nanoparticle (LNP), lipoplex, and / or nanoliposome.

21. The composition comprising the LNP, or The composition of claim 20 , wherein the composition comprises a polymeric carrier, a polycationic protein or peptide, or an artificial RNA associated with the LNP.

22. The LNP comprises: (i) at least one cationic lipid; (ii) neutral lipid; (iii) a steroid or a steroid analogue, and (iv) PEG-lipids It essentially consists of The composition according to any one of claims 20 to 21, wherein (i) to (iv) are, in molar ratio, 20 to 60% cationic lipid, 5 to 25% neutral lipid, 25 to 55% sterol, and 0.5 to 15% PEG-lipid.

23. A polypeptide or any variant of these polypeptides, characterized in that it is encoded by an artificial RNA described in any of claims 1 to 16, having an amino acid sequence identical to or at least 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NOs: 2005, 2743, 3481, 4957, 14309, 15047, 15785, 16523.

24. A composition comprising the polypeptide described in claim 23, optionally comprising at least one pharma- ceutically acceptable carrier or at least one artificial RNA described in any of claims 1 to 16, or a composition described in any of claims 17 to 22.

25. A vaccine comprising an artificial RNA described in any one of claims 1 to 16, a composition described in any one of claims 17 to 22, a polypeptide described in claim 23, or a composition described in claim 24.

26. The vaccine described in claim 25, wherein the artificial RNA described in any of claims 1 to 16, the composition described in any of claims 17 to 22, the polypeptide described in claim 23, or the composition described in claim 24 induces an adaptive immune response.

27. ​​A kit or kit-of-parts comprising an artificial RNA as described in any one of claims 1 to 16, a composition as described in any one of claims 17 to 22, a polypeptide as described in claim 23, or a composition as described in claim 24, and / or a vaccine as described in any one of claims 25 to 26, optionally a liquid vehicle for solubilization, and optionally technical instructions providing information on the dosage and administration of the components.

28. An artificial RNA described in any of claims 1 to 16, a composition described in any of claims 17 to 22, a polypeptide described in claim 23, or a composition described in claim 24, and / or a vaccine described in any of claims 25 to 26, or a kit or kit-of-parts described in claim 27, for use as a pharmaceutical.

29. An artificial RNA described in any of claims 1 to 16, a composition described in any of claims 17 to 22, a polypeptide described in claim 23, or a composition described in claim 24, and / or a vaccine described in any of claims 25 to 26, or a kit or kit-of-parts described in claim 27, for use in the treatment or prevention of a viral infection or a disorder associated with such an infection.

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